3192 ENDO12L_M36A MEET-THE-PROFESSOR: CLINICAL - Clinical Management of Adrenocortical Carcinoma (1:00 PM - 1:45 PM) Clinical Management of Adrenocortical Carcinoma Martin Fassnacht University of Wuerzburg, Wuerzburg, Germany Nothing to Disclose: MF 2012-06-25T13:00:00 Room 320 2012-06-25T00:00:00 1899-12-30T13:00:00 6137 285 1945 M36 A2A Monday 258 2012


3193 ENDO12L_M38A MEET-THE-PROFESSOR: CLINICAL - Evaluation of Premature Adrenarche (1:00 PM - 1:45 PM) Evaluation of Premature Adrenarche Sharon E Oberfield Columbia University Medical Center, New York, NY Nothing to Disclose: SEO 2012-06-25T13:00:00 Room 310 2012-06-25T00:00:00 1899-12-30T13:00:00 6155 286 1946 M38 PED2A Monday 259 2012


3194 ENDO12L_M39A MEET-THE-PROFESSOR: CLINICAL - Pubertal Disruption in Chronic Inflammatory Disease (1:00 PM - 1:45 PM) Pubertal Disruption in Chronic Inflammatory Disease Mark Daniel DeBoer University of Virginia, Charlottesville, VA Disclosure Incomplete: MDD 2012-06-25T13:00:00 Room 360 2012-06-25T00:00:00 1899-12-30T13:00:00 6156 287 1947 M39 PED7A Monday 260 2012


3195 ENDO12L_M40A MEET-THE-PROFESSOR: CLINICAL - Specialized Lipid Testing: The Pre-Treatment Lipoprotein Size Consequences of Adiposopathy ([quot]Sick Fat[quot]) [amp] the Clinical Pitfalls of Post-Treatment Lipoprotein Particle Size Analyses (1:00 PM - 1:45 PM) Specialized Lipid Testing: The Pre-Treatment Lipoprotein Size Consequences of Adiposopathy ([quot]Sick Fat[quot]) [amp] the Clinical Pitfalls of Post-Treatment Lipoprotein Particle Size Analyses Harold Edward Bays L-MARC Research Center, Louisville, KY Disclosures: HEB: Advisory Group Member, Merck & Co.; Speaker, Merck & Co.; Researcher, Merck & Co., Orexigen, Vivus USA, Arena, Novo Nordisk. 2012-06-25T13:00:00 Room 371 2012-06-25T00:00:00 1899-12-30T13:00:00 6222 288 1948 M40 L1A Monday 261 2012


3196 ENDO12L_M41A MEET-THE-PROFESSOR: CLINICAL - What To Do If My Country Doesn[apos]t Have a FRAX? (1:00 PM - 1:45 PM) What To Do If My Country Doesn[apos]t Have a FRAX? Michael R McClung Oregon Osteoporosis Center, Portland, OR Nothing to Disclose: MRM 2012-06-25T13:00:00 Room 361 ABDE 2012-06-25T00:00:00 1899-12-30T13:00:00 6261 289 1949 M41 B11A Monday 262 2012


3197 ENDO12L_CMF9-1A CASE MANAGEMENT FORUM: CLINICAL - Management of Apparent Secondary Hypogonadism Due to Obesity, Opiates [amp] Anabolic Steroids (5:30 PM - 6:15 PM) Management of Apparent Secondary Hypogonadism Due to Obesity, Opiates [amp] Anabolic Steroids Bradley David Anawalt University of Washington, Seattle, WA Disclosure Incomplete: BDA 2012-06-25T17:30:00 Theater A 2012-06-25T00:00:00 1899-12-30T17:30:00 6169 343 2654 CMF9-1 CMF-MR2A Monday 308 2012


3198 ENDO12L_CMF9-2A CASE MANAGEMENT FORUM: CLINICAL - Management of Apparent Secondary Hypogonadism Due to Obesity, Opiates [amp] Anabolic Steroids (5:30 PM - 6:15 PM) Management of Apparent Secondary Hypogonadism due to Obesity, Opiates [amp] Anabolic Steroids Richard Arthur Bebb University of British Columbia, Vancouver, Canada Nothing to Disclose: RAB 2012-06-25T17:30:00 Theater A 2012-06-25T00:00:00 1899-12-30T17:30:00 6182 343 2655 CMF9-2 CMF-MR2A Monday 309 2012


3199 ENDO12L_M42A MEET-THE-PROFESSOR: CLINICAL - Assessing Readiness, Behavioral Modification [amp] Obesity (5:30 PM - 6:15 PM) Assessing Readiness, Behavioral Modification [amp] Obesity Daniel Holland Bessesen University of Colorado School of Medicine, Denver, CO Disclosure Incomplete: DHB 2012-06-25T17:30:00 Room 320 2012-06-25T00:00:00 1899-12-30T17:30:00 6164 344 2656 M42 OB1A Monday 310 2012


3200 ENDO12L_M43A MEET-THE-PROFESSOR: CLINICAL - Common Pitfalls in the Evaluation of Patients with Adrenal Incidentalomas (5:30 PM - 6:15 PM) Common Pitfalls in the Evaluation of Patients with Adrenal Incidentalomas Amir Hekmat Hamrahian Cleveland Clinic Foundation, Cleveland, OH The widespread use of abdominal imaging has resulted in an increased frequency of detected adrenal lesions. An adrenal incidentaloma is an adrenal mass, 1 cm or more in diameter, that is discovered serendipitously by radiological examination in the absence of symptoms or clinical findings suggestive of adrenal disease. When clinicians confront an adrenal incidentaloma, evaluation must address 2 issues: is the lesion functional or is it malignant and if so, is it primary or metastatic. In spite of the fact that the majority of adrenal incidentalomas are benign and non-functional, careful evaluation of all adrenal incidentalomas is warranted to ensure that tumors, such as primary adrenocortical carcinoma (PAC) and pheochromocytoma, that can be associated with significant morbidity and mortality, are not missed.[br]The adrenal computed tomographic (CT) is the cornerstone of imaging studies for adrenal tumors, providing information about size, density, presence of calcifications, areas of necrosis and extent of local invasion of the neoplasm. The three most important imaging criteria are: 1) size of the lesion, 2) the unenhanced CT attenuation value, and 3) the percentage washout. Several studies have shown that the non-contrast CT attenuation is superior to adrenal size in differentiating between benign and malignant adrenal tumors. Appropriate image interpretation can be a guide to a rational diagnostic workup and management.[br]There continues to be disagreement among experts on the choice and the extent of biochemical evaluation in patients with adrenal incidentalomas. All patients with an adrenal incidentaloma should be evaluated for autonomous cortisol secretion, termed subclinical Cushing[apos]s syndrome, (SCS), pheochromocytoma and, if hypertensive, primary aldosteronism. There is significant inconsistency among criteria used in the literature for the diagnosis of SCS. In spite of the lack of a single gold standard test to diagnose SCS, the 1-mg DST is currently considered the best diagnostic tool for evaluation of patients suspected to have SCS. Measurement of plasma metanephrines or 24 hour urine metanephrines are appropriate screening tests for pheochromocytoma. While measurement of plasma aldosterone to renin ratio (ARR) is considered as the best initial screening test for the evaluation of primary aldosteronism, there is a lack of consensus about what ARR cut-off should trigger further evaluation.[br][br]Nothing to Disclose: AHH 2012-06-25T17:30:00 Room 332 2012-06-25T00:00:00 1899-12-30T17:30:00 2459 345 2657 M43 A3A Monday 311 2012


3201 ENDO12L_M44A MEET-THE-PROFESSOR: CLINICAL - Management of Early Stage Cancer/Need for RAI (5:30 PM - 6:15 PM) Management of Early Stage Cancer/Need for RAI Bryan McIver Mayo Clinic, Rochester, MN Session supported by: Genzyme Corporation[br][br]Nothing to Disclose: BM 2012-06-25T17:30:00 Room 310 2012-06-25T00:00:00 1899-12-30T17:30:00 6160 346 2658 M44 TH3A Monday 312 2012


3202 ENDO12L_M45A MEET-THE-PROFESSOR: CLINICAL - Management of Gender Variant Youth (5:30 PM - 6:15 PM) Management of Gender Variant Youth Norman P Spack Children[apos]s Hospital Boston, Boston, MA Nothing to Disclose: NPS 2012-06-25T17:30:00 Room 370 2012-06-25T00:00:00 1899-12-30T17:30:00 6184 347 2659 M45 PED4A Monday 313 2012


3203 ENDO12L_M46A MEET-THE-PROFESSOR: CLINICAL - Minimally Invasive Surgery for Hyperparathyroidism (5:30 PM - 6:15 PM) Minimally Invasive Surgery for Hyperparathyroidism Robert Udelsman Yale University School of Medicine, New Haven, CT Nothing to Disclose: RU 2012-06-25T17:30:00 Room 361 ABDE 2012-06-25T00:00:00 1899-12-30T17:30:00 6208 348 2660 M46 B6A Monday 314 2012


3204 ENDO12L_M48A MEET-THE-PROFESSOR: CLINICAL - Update in FGF 23 (5:30 PM - 6:15 PM) Update in FGF 23 Michael John Econs Indiana University Medical Center, Indianapolis, IN Disclosures: MJE: Indiana University has a patent, but no clinical products at this time, Kirin Brewery; Consultant, Kirin Brewery. 2012-06-25T17:30:00 Room 352 DEF 2012-06-25T00:00:00 1899-12-30T17:30:00 6147 349 2661 M48 B10A Monday 315 2012


3205 ENDO12L_CMF10-1A CASE MANAGEMENT FORUM: CLINICAL - Management of Pediatric Thyroid Cancer (1:00 PM - 1:45 PM) Management of Pediatric Thyroid Cancer Catherine Anne Dinauer Yale University School of Medicine, Guilford, CT Session supported by: Genzyme Corporation[br][br]Nothing to Disclose: CAD 2012-06-26T13:00:00 Theater B 2012-06-26T00:00:00 1899-12-30T13:00:00 6185 389 2783 CMF10-1 CMF-TH2A Tuesday 367 2012


3206 ENDO12L_CMF10-2A CASE MANAGEMENT FORUM: CLINICAL - Management of Pediatric Thyroid Cancer (1:00 PM - 1:45 PM) Management of Pediatric Thyroid Cancer Gary Lee Francis Medical College of Virginia, Richmond, VA Session supported by: Genzyme Corporation[br][br]Nothing to Disclose: GLF 2012-06-26T13:00:00 Theater B 2012-06-26T00:00:00 1899-12-30T13:00:00 6186 389 2784 CMF10-2 CMF-TH2A Tuesday 368 2012


3207 ENDO12L_M49A MEET-THE-PROFESSOR: CLINICAL - Acromegaly (1:00 PM - 1:45 PM) Acromegaly Andrea Lynn Utz Vanderbilt University, Nashville, TN Session supported by: Ipsen Biopharmaceuticals, Inc.[br][br]Disclosures: ALU: Advisory Group Member, Ipsen. 2012-06-26T13:00:00 Room 310 2012-06-26T00:00:00 1899-12-30T13:00:00 6165 390 2785 M49 PIT4A Tuesday 369 2012


2996 ENDO12L_S37-2 SYMPOSIUM SESSION: BASIC - Endocrine Regulation of Lipid [amp] Carbohydrate Metabolism, with Consequences for Longevity: Insights from Animal Models (9:30 AM - 11:00 AM) Regulation of Adult Stem Cell Homeostasis by Metabolic Pathways Anne Brunet Stanford University, Palo Alto, CA Disclosure Incomplete: AB 2012-06-25T10:00:00 Room 342 ABDE 2012-06-25T00:00:00 1899-12-30T10:00:00 6063 261 1852 S37-2 T06-S01 Monday 277 2012


2997 ENDO12L_S37-3 SYMPOSIUM SESSION: BASIC - Endocrine Regulation of Lipid [amp] Carbohydrate Metabolism, with Consequences for Longevity: Insights from Animal Models (9:30 AM - 11:00 AM) Lifespan Extension in the Growth Hormone Receptor Disrupted Mouse John Joseph Kopchick Ohio University/Edison Biotechnical Institute, Athens, OH The growth hormone receptor gene disrupted (GHR-/-) mouse, a.k.a. [apos]The Laron Mouse[apos], was generated in our laboratory nearly two decades ago by targeting exon 4 of the GHR/BP gene. In doing so, both the GHR and GHBP encoded proteins were eliminated. These dwarf mice are insensitive (or resistant) to the action of GH with severely depressed levels of serum IGF-I and elevated levels of GH. GHR-/- mice are fertile; however, at times, breeding is difficult. They also are hypoinsulinemic and insulin sensitive and transition from hypo- to normoglycemic with age. Remarkably, these mice are obese yet long-lived. In terms of obesity, the GHR-/- mice have increased relative fat mass throughout life as compared to littermate controls with a preferential enlargement of the subcutaneous depot. Serum adipokine changes have been noted in these mice with both total adiponectin and high molecular weight (HMW) adiponectin increased. Interestingly, the ratio of the more biologically active HMW adiponectin to total adiponectin is increased in the GHR-/- mice relative to controls. This ratio may be important in the insulin sensitive state found in the mice. Significantly, the GHR-/- mouse mimics many phenotypic aspects of Laron Syndrome patients including depressed incidences of cancer and diabetes. Thus, the use of these mice as a research tool has been widespread and has increased our knowledge of the role of GH in growth, reproduction, longevity, obesity, cancer, insulin and glucose metabolism, the GH/IGF-I feedback loops, and in GH induced intracellular signaling. A review of results derived from these mice along with recent data will be presented.[br][br]The author would like to acknowledge additional authors on this abstract: Darlene Berryman and Ellen Lubbers, Edison Biotechnology Institute, Ohio University, Athens, Ohio.[br][br]Sources of Research Support: This work was supported in part by the State of Ohio[apos]s Eminent Scholar Program that includes a gift by Milton and Lawrence Goll; NIH grant P01AG031736-01A1; and by the AMVETS.[br][br]Nothing to Disclose: JJK 2012-06-25T10:30:00 Room 342 ABDE 2012-06-25T00:00:00 1899-12-30T10:30:00 2469 261 1853 S37-3 T06-S01 Monday 278 2012


2998 ENDO12L_S38-1 SYMPOSIUM SESSION: TRANSLATIONAL - New Insights in the Regulation of Puberty (9:30 AM - 11:00 AM) Lin 28A Transgenic Mice Models: The Human Phenotype Hao Zhu Children[apos]s Hospital Boston and Dana Farber Cancer Institute, Boston, MA Nothing to Disclose: HZ 2012-06-25T09:30:00 Room 362 2012-06-25T00:00:00 1899-12-30T09:30:00 6079 262 1854 S38-1 T07-S06 Monday 280 2012


2999 ENDO12L_S38-2 SYMPOSIUM SESSION: TRANSLATIONAL - New Insights in the Regulation of Puberty (9:30 AM - 11:00 AM) How Many Kisses Are Required To Change a Frog into a Princess? Robert A Steiner University of Washington, Seattle, WA Kisspeptin (a product of the [italic] Kiss1 [/italic] gene) and its receptor (GPR54 or Kiss1r) have emerged as key players in the regulation of reproduction. Mutations in humans and genetically targeted deletions in mice of either [italic] Kiss1 [/italic] or [italic] Kiss1r [/italic] silence the reproductive system, and early developmental expression of Kiss1 or activation of Kiss1r triggers precocious puberty. These observations suggest that kisspeptin signaling is a prerequisite for triggering the onset of puberty. However, recent evidence challenges this notion[mdash] reflecting the discovery that congenital ablation of virtually all ([gt] 97%) of kisspeptin neurons fails to disrupt normal puberty and fertility in mice. Although a few steely-eyed skeptics may question the precise role for kisspeptin in reproduction, others retort that just a few Kiss1-expressing neurons (and the most miniscule amount of Kiss1) may be sufficient to drive puberty and support reproduction. To distinguish between these interpretations and to determine whether males and females differ in their requirement for kisspeptin signaling to achieve reproductive success, we produced a mouse model that has a disruption in Kiss1 transcription ([italic] Kiss1 [sup] Cre/Cre [/sup] [/italic]) and markedly reduced expression of [italic] Kiss1 [/italic]. We found that male [italic] Kiss1 [sup] Cre/Cre [/sup] [/italic] mice exhibit a significant delay in the age of puberty onset and have lower testicular weights than wild type (WT) controls[mdash] however, these same [italic] Kiss1 [sup] Cre/Cre [/sup] [/italic] males can inseminate WT females, which produce normal-sized litters, and retain the ability to show a postcastration rise in gonadotropin secretion. In contrast, female [italic] Kiss1 [sup] Cre/Cre [/sup] [/italic] mice enter puberty at the same age as WT controls but exhibit a dramatically reduced incidence of ovulation and have markedly reduced fertility compared to WTs. Thus, we conclude that either a vanishingly small degree of Kiss1 expression is sufficient to support reproduction or that compensatory mechanisms help to rescue the reproductive phenotype of [italic] Kiss1 [sup] Cre/Cre [/sup] [/italic] mice; moreover, the absolute requirement for Kiss1 to initiate puberty and sustain reproduction in the adult differs between the sexes.[br][br]Sources of Research Support: NIH/NICHD 2 R01 HD049651 to RAS.[br][br]Nothing to Disclose: RAS 2012-06-25T10:00:00 Room 362 2012-06-25T00:00:00 1899-12-30T10:00:00 2532 262 1855 S38-2 T07-S06 Monday 281 2012


3000 ENDO12L_S38-3 SYMPOSIUM SESSION: TRANSLATIONAL - New Insights in the Regulation of Puberty (9:30 AM - 11:00 AM) Genes Implicated in GnRH Secretion Ursula B Kaiser Brigham and Women[apos]s Hospital/Harvard Medical School, Boston, MA The mechanisms controlling the timing and onset of puberty remain largely unknown but recent insights into the genetic causes of pubertal disorders have provided important advances in this field. Mutations in genes important for the development and function of hypothalamic-pituitary pathways controlling GnRH release and LH and FSH secretion have been identified in patients with normosmic hypogonadotropic hypogonadism, Kallmann syndrome, and central precocious puberty. Many of the genes implicated encode G protein-coupled receptors and their ligands, including: (1) [italic]KISS1/KISS1R,[/italic] encoding kisspeptin and its receptor (KISS1R), (2) [italic]TAC3/TACR3[/italic], encoding neurokinin B and the neurokinin 3 receptor (NK3R), (3) [italic]PROK2/PROKR2[/italic], encoding prokineticin 2 and prokineticin receptor 2 (PROKR2), and (4) [italic]GNRH1/GNRHR[/italic], encoding GnRH itself and its receptor, GnRHR. Mutations in these GPCRs, located in diverse functional domains of the receptors, have been described in both heterozygous and homozygous states in patients with varying degrees of GnRH dysregulation. Elucidating the structure-function relationships for these GPCRs and the key mechanisms by which the activation of these receptors by their ligands mediate cellular and biological responses have become increasingly important as reproductive abnormalities are attributed to mutations in these genes. The identification of the domains of these receptors important for cell surface expression, ligand binding, and activation of cellular signaling pathways will advance our understanding of the function of these receptors in the control of GnRH release.[br][br]Nothing to Disclose: UBK 2012-06-25T10:30:00 Room 362 2012-06-25T00:00:00 1899-12-30T10:30:00 2549 262 1856 S38-3 T07-S06 Monday 282 2012


3001 ENDO12L_S39-1 SYMPOSIUM SESSION: CLINICAL - New Insights into Androgen Replacement Therapy (9:30 AM - 11:00 AM) Does Androgen Replacement Drive Prostate Disease? Stephanie T Page University of Washington Medical Center, Seattle, WA Androgen replacement in men with low serum testosterone concentrations can improve symptoms of hypogonadism, such as low libido and reduced sexual function, and has positive effects on body composition and strength. However, concern exists that increases in circulating testosterone concentrations might increase the risk for prostate disease, such as benign prostatic hyperplasia or prostate cancer. Advanced age is the primary, identified risk factor for both diseases of the prostate and hypogonadism and it is likely that these conditions co-exist in significant numbers of men. Unfortunately, randomized-controlled trials appropriately powered to address the relationship, if any, between prostate disease risk and androgen replacement have not been performed. Therefore, in this session we will review the available data addressing separate and important issues regarding testosterone replacement, prostate biology and disease risk.[br]First, do exogenous androgens, acting as a growth factor, drive the normal prostate towards a proliferative or de-differentiated state that might progress to prostate cancer? Androgen replacement can increase prostate size in men with long-standing androgen deficiency. However, recent data suggest that raising serum androgen levels does not increase intraprostatic androgen concentrations in parallel, suggesting that there might be few biologic consequences within the prostate epithelium when serum androgen levels are increased (within the normal range).[br]Second, does androgen replacement in men who have been treated for prostate cancer increase their risk for recurrence? Androgen withdrawal is standard therapy for prostate cancer, and initial responsiveness is essentially universal, suggesting a pivotal role for androgens in maintaining prostate carcinoma once it has developed. Recent reports have challenged the notion that androgens stimulate cancer, citing small series of men who have undergone radical prostatectomy for prostate cancer, developed hypogonadism, and been treated with testosterone without biochemical recurrence. Men with longstanding biochemical remission prior to the initiation of testosterone replacement appear to be less likely to experience increases in PSA. However, these reports must be treated with extreme caution, as controlled clinical trials in this area are lacking. Thus, the true risks and benefits of androgen replacement in men who have been treated for prostate cancer remain largely unknown.[br][br]Disclosures: STP: Principal Investigator, Abbott Laboratories. 2012-06-25T09:30:00 Theater B 2012-06-25T00:00:00 1899-12-30T09:30:00 2489 263 1857 S39-1 T05-S05 Monday 284 2012


3002 ENDO12L_S39-2 SYMPOSIUM SESSION: CLINICAL - New Insights into Androgen Replacement Therapy (9:30 AM - 11:00 AM) When Is Testosterone an Appropriate Treatment for Erectile Dysfunction? Jacques Buvat CETPARP, Lille, France Erectile dysfunction (ED) is prevalent in young hypogonadal men. Some randomized, placebo-controlled trials (RCPTs) demonstrated that testosterone therapy (Tth) normalizes sexual function in such men. This has led to the belief that hypogonadism is an easily curable cause of ED. Serum testosterone (T) is [lt] 10.4 nmol/L in 15% of middle-aged and older men presenting with ED. Observational studies of the effects of Tth on erectile function of such men show limited success rate ([lt] 40%). Several meta-analyses of the RPCTs of Tth also analyzed the effects of this therapy on erectile function of men of any age. A first one, not restricted to men with sexual complaints, concluded to an improvement of erections in men with baseline T [lt] 12 nmol/L. Two further meta-analyses, restricted to men with ED or desire problems, did not confirm the benefit of Tth for erectile function, except in young hypogonadal men. Several reasons may account for such differences in the effects of Tth on erectile function of young and older hypogonadal men with ED. Above all vascular comorbidities are prevalent in older men with ED and late onset hypogonadism (LOH).These may prevent improvement of erections with Tth alone, and may require combination therapy with T and phosphodiesterase type V inhibitor (PDE5I) for optimal results. Moreover ED-associated LOH is often mild or moderate, and may not be the real cause of ED, since 2 population-based studies found that in older men low T is associated with ED, but only below a threshold of 8 nmol/L. In certain cases, the low T associated with ED might be a consequence of reduced sexual activity, as suggested by an increase in T following improvement of erectile function after non-hormonal treatment of ED. Lastly, evidence of requirement of a minimum T level for a complete efficacy of PDE5I therapy recently emerged. The corresponding thresholds seem close to 10.4 nmol/L for total T, 2.1 nmol/L for bioavailable T, and 180 pml/L for calculated Free T. In conclusion, the lower the patient[apos]s age and T level, the higher the chance of success of Tth alone in a hypogonadal patient with ED. Low T is probably a plausible cause of ED only below 8 nmol/L, it but may reduce sexual desire at less decreased levels. After the age of 50, the frequent association of endothelial and vascular factors often requires PDE5I therapy, that should be combined with Tth in case of T [lt] 10.4 nmol/L, due to the need for a minimum T level for a full PDE5I efficacy.[br][br](1) Buvat J, Maggi M, Gooren L, Guay A, Kaufman J, Morgentaler A, Schulman C, Tan H.M, Torres L.O, Yassin A, Zitzmann M. Endocrine Aspects of Male Sexual Dysfunctions. J Sex Med 2010;7:1627-1656. (2) Buvat J, Montorsi F, Maggi M, Porst H, Kaipia A, Colson MH, Cuzin B, Moncada I, Martin-Morales A, Yassin A, Meuleman E, Eardley I, Dean JD, Shabsigh R. Hypogonadal men nonresponders to the PDE5 inhibitor tadalafil benefit from normalization of testosterone levels with a 1% hydroalcoholic testosterone gel in the treatment of erectile dysfunction (TADTEST study). J Sex Med 2011;8:284-293.[br][br]Sources of Research Support: Grant of Bayer Schering Pharma.[br][br]Disclosures: JB: Advisory Group Member, Lilly USA, LLC; Investigator, Bayer Schering Pharma, Johnson & Johnson. 2012-06-25T10:00:00 Theater B 2012-06-25T00:00:00 1899-12-30T10:00:00 2478 263 1858 S39-2 T05-S05 Monday 285 2012


3003 ENDO12L_S39-3 SYMPOSIUM SESSION: CLINICAL - New Insights into Androgen Replacement Therapy (9:30 AM - 11:00 AM) Is Serum Testosterone Concentration in Men Genetically Determined? Claes Ohlsson Institute of Medicine, Sahlgrenska Academy, Gothenburg, Sweden Testosterone is the most important testicular androgen in men. Low serum testosterone concentrations are associated with cardiovascular morbidity, metabolic syndrome, type 2 diabetes mellitus, atherosclerosis, osteoporosis, sarcopenia, and increased mortality risk. Thus, there is growing evidence that serum testosterone is a valuable biomarker of men[apos]s overall health status. Studies in male twins indicate that there is a strong heritability of serum testosterone. We recently perform a large-scale genome-wide association study to examine the effects of common genetic variants on serum testosterone concentrations.[sup]1[/sup] By examining 14,429 men, we show that genetic variants in the sex hormone-binding globulin ([italic]SHBG[/italic]) locus and on the X chromosome are associated with a substantial variation in serum testosterone concentrations and increased risk of low testosterone. Two single-nucleotide polymorphisms at the sex hormone-binding globulin [italic](SHBG)[/italic] locus (17p13-p12) were identified as independently associated with serum testosterone concentration (rs12150660, p=1.2x10[sup]-41[/sup] and rs6258, p=2.3x10[sup]-22[/sup]). Subjects with [ge]3 risk alleles of these variants had 6.5-fold higher risk of having low serum testosterone than subjects with no risk allele. The rs5934505 polymorphism near [italic]FAM9B[/italic] on the X chromosome was also associated with testosterone concentrations (p=5.6x10[sup]-16[/sup]). The reported associations may now be used in order to better understand the functional background of recently identified disease associations related to low testosterone. Importantly, we identified the first known genetic variant, which affects SHBG[apos]s affinity for binding testosterone and the free testosterone fraction and could therefore influence the calculation of free testosterone. This finding suggests that individual-based SHBG-testosterone affinity constants might be required depending on the genotype of this single-nucleotide polymorphism.[br][br]1) [bold][underline]Ohlsson C[/underline][/bold][bold], Wallaschofski, H et al [/bold]2011 Genetic Determinants of Serum Testosterone Concentrations in men. [bold]PloS Genetics[/bold] [italic]Oct;7(10):e1002313[/italic].[br][br]Nothing to Disclose: CO 2012-06-25T10:30:00 Theater B 2012-06-25T00:00:00 1899-12-30T10:30:00 2399 263 1859 S39-3 T05-S05 Monday 286 2012


3004 ENDO12L_S40-1 SYMPOSIUM SESSION: BASIC - New Mechanisms for HPA Axis Regulation (9:30 AM - 11:00 AM) Circadian Clock Genes [amp] Regulation of Adrenal Glucocorticoid Secretion Henrik Oster University of Lubeck, Lubeck, Germany Besides acute activation by stress hypothalamus-pituitary-adrenal (HPA) axis regulation follows strong circadian ([sim]24h period) and ultradian ([lt]24h period) regulation. Under baseline conditions adrenal glucocorticoid (GC) secretion shows a robust circadian rhythm with blood levels peaking around wake-up time (i.e. in the morning in humans and at the beginning of the night in nocturnal rodents). While circadian HPA axis activity is ultimately controlled by the circadian pacemaker residing in the suprachiasmatic nuclei (SCN) of the hypothalamus, different so called peripheral clocks modulate HPA signal propagation at various levels. Circadian oscillators have been described in the paraventricular nucleus, the pituitary and the adrenal cortex. In the latter clock genes regulate the sensitivity of the steroidogenic machinery to ACTH stimulation and, hence, the GC responsiveness during the course of the day. Using a combination of molecular, genetic and surgical approaches we describe the mechanisms of SCN-to-adrenal clock interaction in the regulation of GC rhythms in the mouse and the role of GCs in the synchronization of the circadian machinery in other tissues. Recent data suggest that besides diurnal regulation, clock genes in the adrenal cortex also directly interfere with stress axis activation. In this way, the circadian system might regulate long-term psychological and physiological consequences of repetitive stress exposure.[br][br]Sources of Research Support: German Research Foundation (DFG) Emmy Noether Grant OS 353/4-1; Volkswagen Foundation Lichtenberg Grant.[br][br]Nothing to Disclose: HO 2012-06-25T09:30:00 Room 310 2012-06-25T00:00:00 1899-12-30T09:30:00 1649 264 1860 S40-1 T01-S06 Monday 288 2012


3005 ENDO12L_S40-2 SYMPOSIUM SESSION: BASIC - New Mechanisms for HPA Axis Regulation (9:30 AM - 11:00 AM) Fast Glucocorticoid Feedback Via Endocannabanoid Signaling Jeffrey G Tasker Tulane University, New Orleans, LA Stress leads to activation of corticotropin releasing hormone (CRH) cells in the hypothalamic paraventricular nucleus (PVN) and stimulation of the hypothalamic-pituitary-adrenal (HPA) axis, resulting in an increase in the systemic secretion of glucocorticoids. Glucocorticoids exert both rapid, transient actions and delayed, long-lasting actions in various target tissues throughout the organism, including in multiple areas of the brain. The delayed glucocorticoid effects are mediated by canonical nuclear receptor-mediated transcriptional actions of the steroid, while the rapid effects appear to be mediated by a membrane-associated receptor and cell signaling mechanisms. The acute glucocorticoid actions in the hypothalamus and pituitary gland cause rapid feedback inhibition of the HPA axis. We have investigated the cellular mechanisms responsible for the rapid glucocorticoid feedback inhibition of the HPA axis in a series of in vitro and in vivo studies. We have identified a novel mechanism of rapid glucocorticoid modulation of synaptic signaling in hypothalamic neuroendocrine cells that involves the activation of a membrane glucocorticoid receptor and the rapid synthesis of endocannabinoids in PVN neurons. The retrograde release at glutamate synapses of the endocannabinoid 2-arachidonoylglycerol leads to a presynaptic CB1 receptor-mediated suppression of glutamate release onto the PVN neurons, which causes an inhibition of the excitatory synaptic input to the PVN neurons. The rapid glucocorticoid actions appear to be mediated by the classical intracellular glucocorticoid receptor (GR) located at the membrane, since knocking out the GR causes the loss of the rapid glucocorticoid-induced eCB effect. In vivo, peripheral application of a CB1 antagonist blocked the rapid glucocorticoid suppression of HPA activation induced by restraint stress. Direct infusion of membrane-impermeant glucocorticoid into the PVN inhibited restraint-induced HPA activation, suggesting rapid feedback actions at the PVN. Infusion into the PVN of a CB1 receptor antagonist blocked the rapid glucocorticoid inhibition of the HPA axis. Our findings suggest, therefore, that the rapid glucocorticoid feedback inhibition of the HPA axis is mediated, in part, by the glucocorticoid activation of a membrane-associated GR and the GR-induced retrograde endocannabinoid suppression of the excitatory synaptic drive to the CRH cells.[br][br]Sources of Research Support: NIH grant MH069879; the Catherine and Hunter Pierson Chair in Neuroscience; the Tulane Research Enhancement Fund.[br][br]Nothing to Disclose: JGT 2012-06-25T10:00:00 Room 310 2012-06-25T00:00:00 1899-12-30T10:00:00 2545 264 1861 S40-2 T01-S06 Monday 289 2012


3006 ENDO12L_S40-3 SYMPOSIUM SESSION: BASIC - New Mechanisms for HPA Axis Regulation (9:30 AM - 11:00 AM) Molecular Regulation of CRH Gene Expression Greti Aguilera NIH/NICHD, Bethesda, MD Adequate regulation of hypothalamic corticotropin releasing hormone (CRH) secretion and expression is essential for circadian basal HPA axis activity and for stress adaptation. Transcriptional regulation of the CRH gene depends on cyclic AMP/protein kinase A signaling and binding of phospho-CREB to a CRE at [ndash]270 in the CRH promoter. This CRE is essential for activation of the CRH promoter, and DNA methylation at the internal CpG of this site reduces CREB binding to the promoter affecting CRH expression. However, phospho-CREB alone is not sufficient for driving CRH transcription and emerging evidence indicates that transcriptional activation requires the CREB co-activator, Transducer Of Regulated CREB activity (TORC) and its recruitment by the CRH promoter. In basal conditions, TORC is in the cytoplasm, phosphorylated by the Ser/Thr protein kinases, salt inducible kinase 1 and 2 (SIK1 and 2), and SIK inactivation by PKA allows TORC translocation to the nucleus. Over-expression of TORC1, 2 or 3 potentiates cyclic-AMP-stimulated CRH transcription, while TORC2 and 3 silencing using siRNA blunts it. Immunohistochemistry shows the presence of TORC2 in the cytoplasm of most CRH neurons in basal conditions. Restraint stress causes transient nuclear translocation of TORC2 in about 60% of CRH neurons by 30 min (returning to basal levels by 3 h). Activation of CRH transcription in vitro and in vivo is associated with recruitment of phospho-CREB and TORC2 by the CRH promoter. Increasing evidence indicate that activation/inactivation of TORC in the CRH neuron involves the TORC kinases SIK1 and SIK2. This includes marked induction of SIK1 concomitantly with the declining phase of CRH transcription, and the fact that over-expression of both SIK1 and SIK2 reduces nuclear translocation of TORC and CRH transcription, while the non-selective SIK inhibitor, staurosporin stimulates CRH transcription. Selective silencing of SIK1 or SIK2 using shRNA has revealed differential effects of both isoforms, suggestive that SIK2 is responsible for TORC sequestration in the cytoplasm in basal conditions, while induction of SIK1 probably inactivates TORC in the nucleus and limit CRH transcriptional responses. The overall evidence indicates that TORC is essential for activation of CRH transcription, and suggests that regulation of the SIK/TORC system by stress-activated signaling pathways acts as a sensitive switch mechanism for rapid activation and inactivation of CRH transcription.[br][br]Sources of Research Support: Intramural Research Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development.[br][br]Nothing to Disclose: GA 2012-06-25T10:30:00 Room 310 2012-06-25T00:00:00 1899-12-30T10:30:00 2542 264 1862 S40-3 T01-S06 Monday 290 2012


3007 ENDO12L_S41-1 SYMPOSIUM SESSION: TRANSLATIONAL - Novel Signaling Pathways during Bone Formation (9:30 AM - 11:00 AM) IGF-I Crosstalk with Other Signaling Mechanisms during Bone Formation Daniel D Bikle VA Medical Center 111N-UCSF, San Francisco, CA IGF-I crosstalk with other signaling mechanisms during bone formation.[br]DD Bikle*1 and Y Wang1. Dept Medicine VA Medical Center and University of California San Francisco, San Francsico,CA[br]IGF-I is produced by chondrocytes, osteoblasts, osteocytes, and osteoclasts, and its receptor (IGF-IR) is found in all these cells. The relative contributions of circulating (primarily liver produced) vs paracrine (produced in bone) IGF-I to bone formation is not clear, but both are involved. The actions of IGF-I are mediated by IGF-IR, a tetrameric membrane bound receptor homologous to the insulin receptor. When activated this receptor sets into motion two signaling pathways, the PI3K/Akt pathway and the ras/raf/MAPK/ERK pathway. Akt activation reduces apoptosis by inhibiting BAD, promotes protein synthesis via mTOR, and facilitates [beta]-catenin activation by suppressing GSK3[beta]. Selected integrins are critical for the ability of IGF-I to activate its receptor, modulating the response of bone to IGF-I during skeletal loading and unloading. The global deletion of IGF-I or the chondrocyte specific deletion of IGF-IR leads to a profound reduction in proliferation, increased apoptosis, and blunted differentiation of chondrocytes in the growth plate associated with a reduction in Ihh expression but with increased PTHrP expression presumably accounting for the failure in differentiation. Global deletion of IGF-I or the osteoblast specific deletion of IGF-IR leads to osteopenia and decreased bone formation resistant to the anabolic actions of PTH. Even when IGF-IR is deleted in mature osteoblasts, the ability of PTH to stimulate CFUs (osteoprogenitors) is blocked. Similarly, deletion of IGF-I in osteoblasts and chondrocytes or IGF-IR receptor in osteoclasts blocks osteoclastogenesis and its stimulation by PTH. These results can best be understood by mechanisms involving cell cell signaling. Indeed two such intercellular signaling mechanisms, RANKL/RANK and ephrinB2/EphB4, are dependent for their expression on an intact IGF-I signaling pathway. Disruption of this pathway interferes with the normal regulation of chondrocyte proliferation and differentiation by Ihh/PTHrP and bone remodeling by PTH.[br][br]Sources of Research Support: NIH grants RO1AR055924 and RO1DK054793 awarded to DDB.[br][br]Nothing to Disclose: DDB 2012-06-25T09:30:00 Room 332 2012-06-25T00:00:00 1899-12-30T09:30:00 2463 265 1863 S41-1 T03-S05 Monday 292 2012


3008 ENDO12L_S41-2 SYMPOSIUM SESSION: TRANSLATIONAL - Novel Signaling Pathways during Bone Formation (9:30 AM - 11:00 AM) EphrinB2/EphB4 Interactions Regulating Stromal Cell/Osteoblast Differentiation [amp] Bone Formation Natalie A Sims St Vincent[apos]s Institute of Medical Research, Fitzroy, Australia Ephrins and their receptors (Ephs) are receptor tyrosine kinases that regulate tissue remodeling, including cell migration, angiogenesis and axon guidance. Through direct cell contact, interaction of membrane-bound ephrin ligands with their receptors simultaneously activates signaling through both the receptor (forward) and ligand (reverse). We have reported that ephrinB2 expression by osteoblasts is stimulated by parathyroid hormone (PTH) and its related protein (PTHrP), suggesting endocrine and paracrine regulation of this pathway. In cell culture, specific inhibitors that block signaling of ephrinB2 through EphB4 impaired mineralization by both human and murine osteoblast precursors, and strongly inhibited markers of late stage osteoblast / osteocyte expression. In contrast, they stimulated osteoblastic expression of RANKL, a critical factor for osteoclast differentiation and bone resorption.[br]To study the role of ephrinB2/EphB4 signaling in PTH anabolic action, we treated mice with PTH in the presence and absence of soluble EphB4 (sEphB4) to inhibit ephrinB2/EphB4 signaling. PTH treatment increased trabecular number and thickness, and dramatically increased all osteoblast and bone formation parameters without increasing osteoclast parameters. sEphB4 treatment, with or without PTH, increased osteoblast formation even above PTH-treated levels, but production of osteoid and formation of mineralized bone was not increased, consistent with inhibition of late stage osteoblast markers in vitro. Strikingly, sEphB4 treatment converted PTH anabolic effect to a resorptive response: osteoclast formation was increased and trabecular number reduced. This influence of sEphB4 was confirmed in vitro, where both osteoblastic RANKL production and osteoclast formation in response to PTH or 1,25-dihydroxyvitaminD3 was increased with addition of sEphB4. This enhancing effect of receptor blockade on osteoclast formation might result from the increased RANKL production, but could also relate to sEphB4 preventing ephrinB2 reverse signaling within the osteoclast lineage that has also been reported to limit osteoclast differentiation.[br]These results indicate that ephrinB2/EphB4 signaling enhances late stages of osteoblast differentiation, and communicates with the hemopoietic lineage to restrict osteoclast formation. The increased resorption resulting from ephrinB2/EphB4 receptor blockade needs further assessment in view of the interest in EphB4 blockade as anti-cancer therapy.[br][br]Nothing to Disclose: NAS 2012-06-25T10:00:00 Room 332 2012-06-25T00:00:00 1899-12-30T10:00:00 2455 265 1864 S41-2 T03-S05 Monday 293 2012


3009 ENDO12L_S41-3 SYMPOSIUM SESSION: TRANSLATIONAL - Novel Signaling Pathways during Bone Formation (9:30 AM - 11:00 AM) The Role of Sclerostin in Mediating the Anabolic and Catabolic Effects of PTH in Cortical and Trabecular Bone Alexander G Robling Indiana University School of Medicine, Indianapolis, IN Parathyroid hormone (PTH) is a potent calcium-regulating factor that has skeletal anabolic effects when administered intermittently, or catabolic effects when maintained at consistently high levels. Bone cells express PTH receptors, but the cellular responses to PTH in bone are incompletely understood. Wnt signaling has recently been implicated in the osteo-anabolic response to the hormone. Specifically, the Sost gene, a major antagonist of Wnt signaling, is down-regulated by PTH exposure. We investigated this mechanism by treating Sost-deficient mice and their wild-type littermates with anabolic and catabolic regimens of PTH, and measuring the skeletal responses. Male Sost+/+ and Sost-/- mice were injected daily with human PTH 1-34 (0, 30, or 90 [micro]g/kg) for 6 weeks. Female Sost+/+ and Sost-/- mice were infused with vehicle or high dose PTH (40 [micro]g/kg/day) for 3 weeks. DEXA-derived measures of intermittent PTH (iPTH)-induced bone gain were impaired in Sost-/- mice. Further probing revealed normal or enhanced iPTH-induced cortical bone formation rates but concomitant increases in cortical porosity among Sost-/- mice. Trabecular bone was highly responsive to iPTH in Sost-/- mice. Continuous PTH (cPTH) infusion resulted in equal bone loss in Sost+/+ and Sost-/- mice as measured by DEXA. However, distal femur trabecular bone was spared the bone-wasting effects of cPTH in Sost-/- mice. These results suggest that changes in Sost expression are not required for iPTH-induced anabolism. iPTH-induced resorption of cortical bone might be over-stimulated in Sost deficient environments. Furthermore, Sost deletion protects the trabecular but not cortical compartments from bone loss induced by high-dose PTH infusion.[br][br]Nothing to Disclose: AGR 2012-06-25T10:30:00 Room 332 2012-06-25T00:00:00 1899-12-30T10:30:00 2429 265 1865 S41-3 T03-S05 Monday 294 2012


3010 ENDO12L_S42-1 SYMPOSIUM SESSION: CLINICAL - State of the Art Management of Nodular Thyroid Disease (9:30 AM - 11:00 AM) Application of Molecular Analysis to Clinical Practice Electron Kebebew National Cancer Institute, Bethesda, MD Nothing to Disclose: EK 2012-06-25T09:30:00 Theater A 2012-06-25T00:00:00 1899-12-30T09:30:00 6018 266 1866 S42-1 T02-S03 Monday 296 2012


3011 ENDO12L_S42-2 SYMPOSIUM SESSION: CLINICAL - State of the Art Management of Nodular Thyroid Disease (9:30 AM - 11:00 AM) Follow Up Recommendation for Benign Nodular Disease Edward K Tomimori Sao Paulo University School of Medicine, Sao Paulo, Brazil Disclosure Incomplete: EKT 2012-06-25T10:00:00 Theater A 2012-06-25T00:00:00 1899-12-30T10:00:00 6019 266 1867 S42-2 T02-S03 Monday 297 2012


3256 ENDO12L_M12 MEET-THE-PROFESSOR: CLINICAL - Medical Management of Obesity (2:45 PM - 3:30 PM) Medical Management of Obesity Florencia Halperin Brigham and Women[apos]s Hosp, Boston, MA 2012-06-23T14:45:00 Room 372 2012-06-23T00:00:00 1899-12-30T14:45:00 6303 86 864 M12 OB2 Saturday 12 2012


3257 ENDO12L_M33A MEET-THE-PROFESSOR: CLINICAL - CGMS Sensors: Who Benefits; How [amp] When To Use? (1:00 PM - 1:45 PM) CGMS Sensors: Who Benefits; How & When to Use Joseph Anthony Aloi ' Session supported by: Dexcom, Inc. [amp] Medtronic Diabetes 2012-06-25T13:00:00 Room 352 DEF 2012-06-25T00:00:00 1899-12-30T13:00:00 6288 282 1941 M33 D3A Monday 33 2012


3258 ENDO12L_M12A MEET-THE-PROFESSOR: CLINICAL - Medical Management of Obesity (5:30 PM - 6:15 PM) Medical Management of Obesity Florencia Halperin Brigham and Women[apos]s Hosp, Boston, MA 2012-06-23T17:30:00 Room 372 2012-06-23T00:00:00 1899-12-30T17:30:00 6303 102 902 M12 OB2A Saturday 12 2012


19 ENDO12L_OR04-1 ORAL SESSION: Female Reproduction: The Endometrium [amp] Pregnancy (11:15 AM-12:45 PM) Increased Pregnancy Losses and Poor Neonatal Outcomes in Women with First-Trimester TSH Levels between 2.5 and 4 mIU/L Compared to Euthyroid Women with TSH Less Than or Equal to 2.5 Jubbin Jagan Jacob, Kewal Aditya, Shweta Achint, Tapasya Dhar, Kumkum Avasti Christian Medical College and Hospital, Ludhiana, India; Christian Medical College and Hospital, Ludhiana, India Context: Though maternal and foetal outcomes in overt hypothyroidism have been well studied, data regarding outcomes in subclinical hypothyroidism is conflicting. A recent publication has shown an increase in pregnancy losses among antibody negative women with TSH levels between 2.5 and 5.0mIU/L.(1)[br]Objective: To evaluate pregnancy, maternal and neonatal outcomes in women with first trimester TSH levels [gt] 2.5 but [le] 4mIU/L compared to euthyroid women with TSH [le] 2.5mIU/L irrespective of thyroid antibody levels.[br]Design: The study is part of a prospective study evaluating routine screening of thyroid function tests among pregnant women in the first trimester (Thyroid Screening in Urban Ludhiana Pregnancy Study: TULIPS). Outcomes evaluated include early and late pregnancy losses, 12 pregnancy related maternal complications including preterm delivery, birth weights and 5 neonatal complications. All patients with TSH [gt] 4mIU/L were treated with levothyroxine. For the present study the women were divided into two groups based on the first trimester TSH value, group A with TSH [le] 2.5mIU/L and group B with TSH [gt] 2.5 but [le] 4mIU/L.[br]Setting: The study was conducted in the antenatal clinic of a university affiliated teaching hospital.[br]Patients: Of the 1000 patients recruited, data was available for 951, of whom 790 delivered/aborted in the study hospital and in another 161 patients; the outcomes were obtained over the telephone.[br]Results: There were 533 patients in group A and 263 in group B. There was a significant increase in early ([le] 20 wks.) and late ([gt] 20 wks.) pregnancy losses in group B vs. group A (Early; 8.4% vs. 4.1% p[lt]0.001 [OR (95%CI) 2.1(1.1-3.9)], Late; 4.2% vs. 0.6% p[lt]0.001 [OR (95%CI) 7.7(2.1-27.8)] and combined 12.5% vs. 4.7%, p[lt]0.001[OR (95%CI) 2.9(1.6-5.0)]). The odds (95%CI) of preterm delivery ([lt]37wks.) in group B was 2.02(1.1-3.7) compared to group A. There were no statistically significant differences between the two groups among the other 11 maternal outcomes. Neonates born to women in group B had significantly higher odds of being LBW (24.2% vs. 16.6% p=0.01 [OR (95%CI) 1.6(1.1-2.3)]), SGA (18.6% vs. 10.7%, p[lt]0.001 [OR (95%CI) 1.9(1.2-2.9)]) and had more instances of neonatal hyperbilirubinemia (12.1% vs. 7.5% p=0.04).[br]Conclusions: The clear cut increase in pregnancy losses with TSH [gt] 2.5 but [le] 4mIU/L and significant adverse neonatal outcomes provides another evidence for lowering the TSH range in pregnant women.[br][br]Negro R et al.,J Clin Endocrinol Metab 2010;E44-48.[br][br]Nothing to Disclose: JJJ, KA, SA, TD, KA 2012-06-23T11:15:00 352DEF 2012-06-23T00:00:00 1899-12-30T11:15:00 970 37 74 OR04-1 OR200-01 Saturday 19 2012


20 ENDO12L_OR04-2 ORAL SESSION: Female Reproduction: The Endometrium [amp] Pregnancy (11:15 AM-12:45 PM) Evidence for Bone Marrow Origin of Endometrial Cells in a Murine Model Sara S Morelli, Pranela Rameshwar, Laura T Goldsmith New Jersey Medical School of UMDNJ, Newark, NJ; New Jersey Medical School of UMDNJ, Newark, NJ Mechanisms used in cyclic regeneration of endometrial cellular compartments are unknown. Recent evidence supports the existence of resident stem cell population(s) which give rise to mature endometrial parenchymal cell types. Their origin is unknown. Few studies have attempted to determine the cellular origin of endometrial cells. Current studies determined whether bone marrow (BM) is a source of endometrial epithelial and/or stromal cells.[br]BM cells, harvested from Green Fluorescent Protein (GFP) transgenic donor mice, were injected into lethally irradiated, immature female mice (two independent expts, n=7 or 8 recipients in each). Flow cytometry was used to detect GFP positive cells in peripheral blood of recipient mice. Vaginal cytology determined whether irradiated mice were undergoing estrus cycles. Four animals with successful BM reconstitution, confirmed to be cycling, were sacrificed at either 5 months (n=2, 1 from each expt) or 9 months (n=2, 1 from each expt) post transplant, and hysterectomy was performed. Uteri were assessed by confocal laser microscopy to identify and localize GFP positive cells and fluorescently stained nuclei in recipient uterine tissue sections. Cell counting was performed using computer-assisted image analysis. Data are expressed as GFP positive cell number/total number of stromal compartment cells.[br]5/8 (Expt 1) and 6/7 (Expt 2) recipient mice were successfully reconstituted with donor BM at 7-8 weeks post transplant. In the 11 reconstituted animals, the range of GFP positive peripheral blood cells was 81.3 to 92.2%. At 5 months post transplant, 5.5% of cells (148/2690, Expt 1, n=1 animal) and 3.6% of cells (95/2659, Expt 2, n=1) in the endometrial stromal compartment were GFP positive. At 9 months post transplant, 21.5% of cells (584/2721, Expt 1, n=1) and 18.2% of cells (305/1678, Expt 2, n=1) in the endometrial stromal compartment were GFP positive. In distinct contrast, no endometrial glandular or luminal epithelial cells were GFP positive.[br]These data demonstrate that bone marrow is a significant source of endometrial cells in the stromal, but not epithelial, compartment. The contribution of bone marrow derived-cells to the endometrial stromal compartment increases with time/with increasing number of estrus cycles. These novel data provide an important advance in our understanding of the cellular source of regenerative cells of the endometrium.[br][br]Sources of Research Support: American Board of Obstetrics and Gynecology/American Association of Obstetricians and Gynecologists Foundation Research Training Scholarship; American Society for Reproductive Medicine Career Development Award in Reproductive Medicine and Biology.[br][br]Nothing to Disclose: SSM, PR, LTG 2012-06-23T11:30:00 352DEF 2012-06-23T00:00:00 1899-12-30T11:30:00 1745 37 75 OR04-2 OR200-01 Saturday 20 2012


21 ENDO12L_OR04-3 ORAL SESSION: Female Reproduction: The Endometrium [amp] Pregnancy (11:15 AM-12:45 PM) Absence of Kr[uuml]ppel-Like Factor 9 (KLF9) Promotes Ectopic Endometrial Lesion Development in a Mouse Model of Endometriosis Melissa E Heard, Christian D Simmons, Kim E Light, Frank A Simmen, Rosalia CM Simmen University of Arkansas for Medical Sciences, Little Rock, AR; University of Arkansas for Medical Sciences, Little Rock, AR; Arkansas Children[apos]s Research Institute, Little Rock, AR Endometriosis is a benign gynecological disorder commonly associated with uterine progesterone (P) -resistance and reduced fertility. Kr[uuml]ppel-like factors (KLF) are zinc finger-containing transcription factors which regulate diverse physiological processes in the female reproductive tract. Our group has identified KLF9 as a transcriptional co-regulator of estrogen and P receptor (PGR) signaling in endometrial stroma and glandular epithelial cells [italic]in vitro[/italic]. We recently demonstrated that [italic]KLF9[/italic] mRNA levels are decreased in eutopic endometria of women with vs. without endometriosis. Mice null for [italic]Klf9[/italic] are sub-fertile due to decreased numbers of implantation sites and exhibit attenuated P-sensitivity of uterine endometrium. To determine a causal relationship between P-resistant endometrium due to loss of KLF9 transcriptional control and predisposition to endometriosis, we utilized a mouse model of endometriosis induced by intraperitoneal (IP) injection of minced donor endometrial tissue into immune competent recipient mice. Initial studies with endometria from intact mice expressing enhanced green fluorescent protein administered IP into mice of the same strain confirmed the formation of peritoneal ectopic lesions. To compare predisposition of [italic]Klf9[/italic]-expressing (WT) and [italic]Klf9[/italic] null (KO) endometria for ectopic lesion development, WT recipient mice (8-10 weeks) in the estrous phase were administered IP with minced endometrium ([sim]40 mg tissue in 400[micro]l PBS) from WT or KO mice (10-12 weeks, estrus phase) or with PBS alone (sham control). Eight weeks later, recipient mice (n=10 and 9, respectively for WT and KO donors; one donor to one recipient) were analyzed for presence, size, and histology of endometrial lesions and for body weight changes pre- and post-endometriosis. While 50% of mice injected with donor WT tissue developed detectable lesions, all recipients of donor KO endometrium showed established peritoneal lesions. Donor WT and KO endometrial tissues generated similar numbers of lesions in WT recipient mice; however, lesions from KO donors were larger than those from WT donors (30.3[plusmn]11.3 vs. 19.8[plusmn]12.0 mm[sup]3[/sup]). Mice with and without ectopic lesions did not differ in body, uterine or ovarian weights. Eutopic endometria of WT mice with ectopic lesions did not differ in expression levels of [italic]Pgr[/italic], [italic]Igfbp1[/italic], [italic]Wnt4[/italic] and[italic] Wnt2[/italic] as a function of donor source at sac. Our findings suggest that endometrial KLF9 deficiency maybe causal to the etiology of human endometriosis.[br][br]Sources of Research Support: NIH-HD21961 and ABI-ACHRI Student and Clinical Staff Intramural Grants.[br][br]Nothing to Disclose: MEH, CDS, KEL, FAS, RCMS 2012-06-23T11:45:00 352DEF 2012-06-23T00:00:00 1899-12-30T11:45:00 324 37 76 OR04-3 OR200-01 Saturday 21 2012


22 ENDO12L_OR04-4 ORAL SESSION: Female Reproduction: The Endometrium [amp] Pregnancy (11:15 AM-12:45 PM) A Novel Role for Oviductal Epithelial Estrogen Receptor [alpha] during Fertilization and Early Embryo Development Wipawee Winuthayanon, Sylvia C Hewitt, Elizabeth Padilla-Banks, Grant D Orvis, Richard R Behringer, Carmen J Williams, Kenneth S Korach National Institute of Environmental Health Sciences (NIEHS)/National Institute of Health (NIH), Research Triangle Park, NC; Sloan-Kettering Institute, New York, NY; University of Texas MD Anderson Cancer Center, Houston, TX Loss of epithelial estrogen receptor [alpha] (ER[alpha]) from the uterus causes female infertility, in part, due to an implantation defect. We hypothesized that loss of oviductal epithelial ER[alpha] also contributed to their infertility. By flushing embryos at various times after mating, we showed that epithelial ER[alpha] conditional knockout (cKO) mice lost all of their embryos between days 1 and 2 of pregnancy. Although similar numbers of oocytes were ovulated, only [sim]50% of oocytes collected from the cKO oviducts were fertilized when compared to control littermates, and no 2-cell embryos were ever recovered on pregnancy day 2. The fertilization rate of the cKO oocytes was restored to that of control littermates only when the oocytes were collected directly from the ovaries prior to ovulation and then fertilized in vitro Furthermore, when wild type zygotes were transferred into oviducts of pseudopregnant cKO mice, the majority of the embryos were under-developed when flushed from the oviduct at 3.5 dpc, and no embryos were found in the uterus. In contrast, all transferred embryos collected from pseudopregnant control littermates were morulas or blastocysts, and all of these embryos had reached the uterus. These findings suggest that ablation of ER[alpha] in the oviductal epithelium results in alterations in the oviductal microenvironment that completely disrupts fertilization, preimplantation embryo development and embryo transport. Microarray analysis of cKO and control oviducts collected at 0.5 and 1.5 dpc demonstrated significant alterations in expression of genes involved in interleukin 17 signaling and prostaglandin D2 synthesis. These findings were confirmed at the protein level by analysis of IL17 and prostaglandin D synthase. It can be concluded that epithelial ER[alpha] is required to establish the appropriate oviduct environment for successful mouse preimplantation embryo development. Our findings demonstrate a highly novel and unexpected regulatory phenomenon of epithelial ER[alpha] in the female reproductive tract that has the potential to provide a better understanding of idiopathic infertility in women.[br][br]Nothing to Disclose: WW, SCH, EP-B, GDO, RRB, CJW, KSK 2012-06-23T12:00:00 352DEF 2012-06-23T00:00:00 1899-12-30T12:00:00 988 37 77 OR04-4 OR200-01 Saturday 22 2012


23 ENDO12L_OR04-5 ORAL SESSION: Female Reproduction: The Endometrium [amp] Pregnancy (11:15 AM-12:45 PM) Bone Morphogenic Protein Receptor Type 2 (BMPR2) Is Required for Establishment of an Adequate Uterine Environment To Support Normal Placentation Takashi Nagashima, Qinglei Li, John P Lydon, Francesco J DeMayo, Martin M Matzuk Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Texas A[amp]M University, College Station, TX Bone morphogenetic proteins (BMPs), known to play a critical role in reproductive function, are members of transforming growth factor-[beta] (TGF-[beta]) superfamily and regulate a variety of cellular functions. BMP receptor type 2 (BMPR2) transduces BMPs signals by forming heteromeric complexes with BMP type 1 receptors. Of note, mutant embryos homozygous for a [italic]Bmpr2[/italic] null allele ([italic]Bmpr2[sup]-/-[/sup][/italic]) die during gastrulation. To overcome the early embryonic lethality and understand the [italic]in vivo[/italic] roles of BMPR2 in reproduction, we used progesterone receptor Cre ([italic]Pgr-Cre[/italic]) to conditionally disrupt the [italic]Bmpr2[/italic] gene in the female reproductive tract. [italic]Bmpr2[/italic] conditional knockout ([italic]Bmpr2[/italic] cKO: [italic]Bmpr2[sup]flox/-[/sup]Pgr[sup]cre/+[/sup][/italic]) female mice are complete infertile. However, the ovarian function remains normal, as evidenced by normal ovarian morphology, follicle growth, cumulus expansion, ovulation, oocyte fertilization, and hormone levels, indicating the infertility of [italic]Bmpr2[/italic] cKO mice arises from uterine dysfunction. [italic]Bmpr2[/italic] cKO uteri showed reduced uterine weight after artificial decidualization, which likely results from defective proliferation and differentiation of uterine stromal cells and reduction of blood vessel diameter. However, implantation and early embryo development occurred normally in [italic]Bmpr2[/italic] cKO mice, as shown by normal embryo morphology and weight, number of implantation sites, and implantation- and trophoblast-specific-gene expressions. Strikingly, [italic]Bmpr2[/italic] cKO mice showed severe hemorrhage in trophoblast and decidual tissues of postimplantation uteri at the initiation of vascular remodeling. This defect was accompanied by multiple abnormalities, leading to fetal-growth retardation and loss of the fetus before birth. These results indicate that BMPR2 is indispensable for the establishment of an adequate uterine environment to support normal placentation in mice.[br][br]Sources of Research Support: NIH Grant HD32067 from the Eunice Kennedy Shriver National Institute of Child Health and Human Development.[br][br]Nothing to Disclose: TN, QL, JPL, FJD, MMM 2012-06-23T12:15:00 352DEF 2012-06-23T00:00:00 1899-12-30T12:15:00 1069 37 78 OR04-5 OR200-01 Saturday 23 2012


24 ENDO12L_OR04-6 ORAL SESSION: Female Reproduction: The Endometrium [amp] Pregnancy (11:15 AM-12:45 PM) Transcription Factor ZEB1 Acting Via Specific microRNAs (miRNA/miR) Plays a Pivotal Role in Hormonal Control of Uterine Quiescence and Contractility during Pregnancy and Labor Koriand[apos]r Williams, Nora E Renthal, Carole R Mendelson University of Texas Southwestern Medical Center, Dallas, TX Uterine quiescence is maintained during pregnancy by progesterone (P[sub]4[/sub]) acting via its nuclear receptors (PR) to inhibit expression of genes encoding contraction-associated proteins (CAPs) ([italic]e.g[/italic]. oxytocin receptor (OTR), connexin-43 (CX43) and cyclooxygenase-2 (COX-2)) in the myometrium. This occurs, in part, via P[sub]4[/sub]/PR anti-inflammatory actions. On the other hand, parturition is associated with a decline in PR function, resulting in an increased inflammatory response within the myometrium. Moreover, circulating estradiol (E[sub]2[/sub]) levels and/or estrogen receptor [alpha] activation, which increase markedly near term, contribute to the uterine inflammatory response and antagonize the anti-inflammatory actions of P[sub]4[/sub]/PR. Previously, we found that during the transition to labor the decline in PR function resulted in decreased expression of ZEB1/2, which allowed for the induction of miR-200 expression and increased transcription of OTR and CX43. Interestingly, we recently identified a conserved cluster of miRNAs, miR-199a and miR-214, that were significantly downregulated in pregnant mouse and human myometrium near term and during labor, whereas, their target, COX-2, a critical enzyme in the production of contractile prostaglandins, was coordinately increased. This pattern of expression also was observed in an inflammatory (lipopolysaccharide-induced) mouse model of preterm labor. Physiological relevance of these miRNAs was suggested by the finding that overexpression of miR-199a-3p/214 mimics in cultured human myometrial cells inhibited COX-2 protein expression and blocked TNF-[alpha]-induced myometrial cell contractility. Notably, E[sub]2[/sub] treatment of ovariectomized mice suppressed uterine miR-199a-3p/214 expression and coordinately induced COX-2 protein, whereas P[sub]4[/sub] treatment enhanced miR-199a-3p/214 expression and repressed COX-2. Intriguingly, these opposing hormonal effects were mediated by ZEB1, which we found to activate miR199a/214 expression. Remarkably, ZEB1 was inhibited by E[sub]2[/sub] and stimulated by P[sub]4[/sub] in uterine tissues of ovarectomized mice. Consequently, binding of endogenous ZEB1 to the miR-199a/214 promoter was found to be diminished in the pregnant mouse uterus at term. Furthermore, in co-transfected COS-7 cells, ZEB1 overexpression induced miR-199a/214 promoter activity. Our collective findings point to the key pivotal roles of myometrial ZEB1 and its miRNA targets as hormonally-controlled regulators of inflammatory and contractile gene expression in the pregnant uterus.[br][br]Sources of Research Support: P01-HD-11149, Prematurity Research Initiative Grant (#21FY11), March of Dimes Foundation, Pharmacological Sciences Training Grant.[br][br]Nothing to Disclose: KW, NER, CRM 2012-06-23T12:30:00 352DEF 2012-06-23T00:00:00 1899-12-30T12:30:00 1678 37 79 OR04-6 OR200-01 Saturday 24 2012


25 ENDO12L_OR05-1 ORAL SESSION: Hormone Dependent Tumors (11:15 AM-12:45 PM) Mitochondrial Estrogen Receptor Beta Dictates Tamoxifen Resistance or Sensitivity in Breast Cancer Ali Pedram, Mahnaz Razandi, Suzanne Fuqua, Ellis R Levin University of California, Irvine, CA; Long Beach VA Medical Center, Long Beach, CA; Baylor College of Medicine, Houston, TX Tamoxifen (TAM) resistance in breast cancer occurs in approximately 50% of women. TAM[apos]s effectiveness importantly involves a cyto-toxic action and underlies responses to TAM. We investigated several TAM sensitive or resistant cell lines most notably MCF7-BK (sensitive) or MCF7 BK-TR (resistant) cells. At 24 hours, TAM induced strong mitochondrial ROS generation in BK cells that caused a 13-fold increase in apoptosis compared to no TAM. Increased ROS and apoptosis resulted from 70% decreased MnSOD activity, a mitochondrial enzyme that processes superoxide ROS. Decreased mitochondrial MnSOD activity was due to TAM inducing nitric oxide-dependent nitrosylation at tyrosine 34 of MnSOD, by our mutational analysis required for inhibition of MnSOD activity. TAM-generated ROS induced PKCdelta and JNK kinase activation, BAX translocation to mitochondria, and the TAM-induced secretion of cytochrome c (apoptosome component) directly from isolated mitochondria. These functions in BK cells resulted from TAM acting as an ERbeta antagonist, shown by siRNA knockdown of ERbeta or ERalpha, and opposition to TAM by DPN (ERbeta agonist) but not PPT (ERalpha agonist). ERbeta was mainly localized to the mitochondria of the cells by fluorescent microscopy and sub-cellular immuno-blotting. In contrast, TAM acted as an ERbeta agonist in BK-TR (resistant) cells, decreasing nitrosylation to stimulate MnSOD activity, diminishing ROS levels and preventing apoptosis. Knocking down MnSOD with siRNA resulted in reversal of resistance to TAM in BK-TR cells, increasing cell death 15-fold. BK-TR tumors were grown in both flanks of nude mice (n=6) under TAM pellet administration for 5 weeks to 1200mm[sup]3[/sup]on average. Tumors were then injected with either lentiviral control or MnSOD shRNAs. MnSOD knockdown was seen only in the latter tumors that substantially regressed to 480mm[sup]3[/sup] at 8 weeks while control shRNA-injected tumors grew to 1690mm[sup]3[/sup], all in the presence of TAM. Caspase 7 was cleaved and apoptosis was confirmed (Annexin V histological staining) only in the MnSOD shRNA-injected tumors. MnSOD lentiviral shRNA in matrigel at initiation almost completely prevented in-vivo tumor formation over 5 weeks, compared to control shRNA and BK-TR cells in matrigel. Thus, mitochondrial ERbeta serves as an ROS rheostat to determine breast tumor fate. Inhibiting the tumor MnSOD and engaging ERbeta as antagonist overcomes TAM resistance and promotes apoptosis in human breast cancer.[br][br]Sources of Research Support: NCI Grant CA100366 (ERL).[br][br]Nothing to Disclose: AP, MR, SF, ERL 2012-06-23T11:15:00 342ABDE 2012-06-23T00:00:00 1899-12-30T11:15:00 24 38 80 OR05-1 OR42-01 Saturday 25 2012


26 ENDO12L_OR05-2 ORAL SESSION: Hormone Dependent Tumors (11:15 AM-12:45 PM) SLIRP Represses Retinoic Acid, SOX9 and Notch Signaling in Colorectal Cancer and Is a Good Prognostic Factor Patrick Aaron Candy, Michael Phillips, Andrew Redfern, Lisa Stuart, John A Davidson, Barry Iacopetta, Ben Wood, Shane M Colley, Peter J Leedman University of Western Australia, Western Australian Institute for Medical Research (WAIMR), Perth, Australia; University of Western Australia, Perth, Australia; Royal Perth Hospital, Perth, Australia; Sir Charles Gairdner Hospital and QEII Medical Centre, Perth, Australia; University of Western Australia, Perth, Australia Colorectal cancer (CRC) is the second highest cause of cancer death and the third most common malignancy. Aberrant Notch signaling in CRC promotes epithelial to mesenchymal transition, chemoresistance and metastasis. Its effects are mediated through transcription factors (TFs), in particular HES1 and HEY1, which influence COX2, NF-[kappa]B and Wnt signaling. In CRC, Notch signaling can be activated by retinoic acid, through the nuclear receptor (NR) RAR[alpha] and the early developmental TF SOX9, which activates HES1. Using CRC tissue microarrays (TMAs), we found that SLIRP, a nuclear receptor (NR) corepressor, is a good prognostic factor in CRC, with a tumor suppressor phenotype. High tumor SLIRP expression in a TMA cohort of 967 patients correlated with improved 5 year survival (p[lt]0.01) and inversely with tumor stage and lymph node invasion. In two CRC cDNA microarray sets, high SLIRP mRNA expression correlated with improved disease free survival over three years following surgery (p[lt]0.05). Here we investigated the mechanism for this clinical advantage and the hypothesis that SLIRP is a repressor Notch signaling. In human CRC cell transfection studies using luciferase (Luc) reporters, siRNA mediated depletion of SLIRP increased the activity of ligand-stimulated RAR[alpha], HES1 and SOX9 signaling, resulting in increased expression of downstream targets, including NOTCH2, NOTCH3, NFKB1, NFKB2, LMO2, HES1 and SOX9. In ChIP studies, SLIRP was recruited (with RAR[alpha]) to the HES1 and SOX9 promoters. When SLIRP was depleted from the cells with siRNA, ChIP studies revealed an increase in RAR[alpha] recruitment to the HES1 and SOX9 promoters, along with increased binding of SOX9 to the HES1 promoter, and a decrease in binding of HES1 to its own promoter. Further, we found that siRNA mediated SLIRP depleted CRC cells were more invasive in matrigel assays and more resistant to the standard CRC chemotherapeutic agents, 5-Fluorouracil and irinotecan. Taken together, these data suggest that SLIRP is a potent suppressor of Notch and RAR signaling in CRC and provide insight into the mechanisms by which SLIRP functions as a tumor suppressor to reduce invasion and enhance sensitivity to chemotherapy in this disease.[br][br]Nothing to Disclose: PAC, MP, AR, LS, JAD, BI, BW, SMC, PJL 2012-06-23T11:30:00 342ABDE 2012-06-23T00:00:00 1899-12-30T11:30:00 1055 38 81 OR05-2 OR42-01 Saturday 26 2012


27 ENDO12L_OR05-3 ORAL SESSION: Hormone Dependent Tumors (11:15 AM-12:45 PM) The Prostate Cancer Specific TMPRSS2:ERG (T/E) Fusion Cooperates with the Vitamin D Receptor To Induce CYP24A1, the Enzyme Responsible for Inactivation of 1,25-Dihydroxyvitamin D[sub]3[/sub] Jung-Sun Kim, Justin Michael Roberts, William Edward Bingman, Nancy Lynn Weigel Baylor College of Medicine, Houston, TX Results of epidemiological and clinical studies assessing the potential benefit of vitamin D in prostate cancer (PCa) have been conflicting even though pre-clinical studies in PCa models have shown that activation of the vitamin D receptor (VDR) reduces prostate cell proliferation and tumor growth. Although the majority of human PCas express a TMPRSS2:ETS factor fusion gene (T/E) which links the androgen regulated promoter of TMPRSS2 to the coding regions of ETS transcription factors (most commonly ERG), most pre-clinical studies have been performed in models lacking T/E. The level of the active metabolite of vitamin D, 1,25-dihydroxyvitamin D[sub]3[/sub] (1,25D), is controlled in part by VDR dependent induction of CYP24A1 which inactivates 1,25D. Since ETS factors can cooperate with VDR to induce CYP24A1, we hypothesized that T/E would cause aberrant induction of CYP24A1 limiting the beneficial effects of VDR action. In VCaP cells, ERG specific siRNAs reduced T/E to 30% of control and reduced 1,25D mediated induction of CYP24A1 to less than 10% of control. In contrast, 1,25D dependent induction of another vitamin D target gene (TRPV6) was not reduced. Induction of CYP24A1 by 1,25D was greatly increased in an LNCaP cell line that overexpresses a T/E isoform (Type VI +72). In T/E overexpressing LNCaP cells, 1 nM 1,25D caused an induction of CYP24A1 equivalent to 100 nM 1,25D in control LNCaP cells. T/E expression also increased maximal CYP24A1 expression at optimal levels of 1,25D suggesting that ETS factors are limiting in cells lacking T/E. Thus, one of the early effects of T/E in prostate cells is a reduction in intracellular 1,25D, which may lead to increased proliferation. Our ChIP analyses showed that ERG bound to the CYP24A1 promoter region predicted to contain both VDR and ETS factor binding sites. Preliminary data show that 1,25D enhances the extent of ERG binding suggesting an interaction between the factors. YK-4-279, an ERG inhibitor, did not reduce 1,25D mediated CYP24A1 induction in VCaP cells. The mechanism by which YK-4-279 inhibits ERG activity in PCa cells is unknown, but it does not prevent DNA binding and cannot prevent cooperation with VDR. Thus, although this compound would be beneficial in counteracting other activities of T/E, it would not prevent the aberrant induction of CYP24A1 by T/E. These new findings suggest that presence of T/E may reduce beneficial actions of VDR by increasing expression of CYP24A1 which inactivates 1,25D.[br][br]Nothing to Disclose: J-SK, JMR, WEB, NLW 2012-06-23T11:45:00 342ABDE 2012-06-23T00:00:00 1899-12-30T11:45:00 1127 38 82 OR05-3 OR42-01 Saturday 27 2012


28 ENDO12L_OR05-4 ORAL SESSION: Hormone Dependent Tumors (11:15 AM-12:45 PM) Macrophage-Derived Amphiregulin Causes Estrogen Receptor-[alpha] Down-Regulation in Tamoxifen-Resistant Breast Cancer Cells Fabio Stossi, Zeynep Madak-Erdogan, Rosa Ventrella, Benita S Katzenellenbogen Baylor College of Medicine, Houston, TX; University of Illinois at Urbana-Champaign, Urbana, IL Breast cancer is the second leading cause of death in US women. Approximately 70% of primary breast tumors express the estrogen receptor alpha (ER), which is routinely used as a prognostic marker and is pharmacologically targeted by endocrine therapies (e.g. tamoxifen or aromatase inhibitors). However, up to 1/3 of women that receive tamoxifen treatment will have a recurrence that has become resistant to the therapy. In this study, we wanted to follow up on our previous observation (1) that macrophage-derived factors can cause down-regulation of ER expression, thus directly impinging on the efficacy of endocrine therapies. We therefore analyzed the actions of macrophages on breast cancer cells that are resistant to tamoxifen treatment because relatively little is known about the role of elements of the tumor microenvironment in the development of endocrine resistance.[br]In this study, we found that macrophage derived factors caused ER down-regulation also in tamoxifen resistant breast cancer cells. Interestingly this happened under different macrophage polarization conditions when compared with tamoxifen sensitive cells. In resistant cells, conditioned media from unpolarized or M2 (pro-tumoral)-like macrophages was responsible for ER[alpha] loss while M1 (anti-tumoral)-like caused this in tamoxifen sensitive cells. By analyzing macrophage conditioned media activated pathways in tamoxifen resistant cells; we found that ER[alpha] down-regulation was mediated via an EGFR-dependent mechanism. Through protein arrays, recombinant factor treatments and antibody-depletion studies, we identified amphiregulin as the main EGFR ligand that elicits reduction of ER[alpha] levels in breast cancer cells. Of note, our findings revealed that this amphiregulin/EGFR paracrine, not autocrine, loop switches the growth potential of tamoxifen resistant breast cancer cells from ER-dependent to EGFR-dependent. No effect of amphiregulin or EGFR inhibitors was seen in tamoxifen sensitive cells. These observations further validate the importance of understanding which factors are present in the tumor microenvironment that will directly impinge on tumor cells, altering the signaling and growth milieu while directly contributing to the success or failure of therapeutic approaches.[br][br](1)Stossi, F., Madak-Erdogan, Z., and Katzenellenbogen, B.S. Macrophage-derived Soluble Factors Cause Loss of Estrogen Receptor Alpha in Breast Cancer Cells via Hyper-activation of ERK2 and c-Jun. Oncogene in press, PMID: 21860415.[br][br]Sources of Research Support: Grant from The Breast Cancer Research Foundation (BSK).[br][br]Nothing to Disclose: FS, ZM-E, RV, BSK 2012-06-23T12:00:00 342ABDE 2012-06-23T00:00:00 1899-12-30T12:00:00 1250 38 83 OR05-4 OR42-01 Saturday 28 2012


29 ENDO12L_OR05-5 ORAL SESSION: Hormone Dependent Tumors (11:15 AM-12:45 PM) Role of microRNA-141 in Hormone-Dependent Breast Cancer Jessica Finlay-Schultz, Diana M Cittelly, Peter Hendricks, Purvi R Patel, Dawn M Cochrane, Britta M Jacobsen, Jennifer K Richer, Carol A Sartorius University of Colorado Denver Anschutz Medical Campus, Aurora, CO; University of Colorado Denver Anschutz Medical Campus, Aurora, CO MicroRNAs (miRNAs) are involved in many aspects of cancer biology, including tumor progression, chemoresistance, and metastasis. In breast cancer specifically, miRNAs are more abundant in well differentiated cancer cells, and many are downregulated or lost in more de-differentiated cancer cells. Luminal breast cancers are estrogen receptor (ER) and progesterone receptor (PR) positive. These cancers are hormone dependent and require estrogens for growth; the role of progesterone is less clear. Progesterone is known to rapidly downregulate the expression of several miRNAs in breast cancer cell lines. This includes miR-141, a member of the miR-200 family, of which miR-200c is a well-known tumor suppressor. Interestingly, of the miR-200 family, only miR-141 was found to be significantly hormone regulated in breast cancer cells, and is also reported to be downregulated in breast cancer stem cells (CSCs). Therefore, we hypothesize that loss of miR-141 plays a role in triggering expansion of the CSC population in breast tumors.[br]Preliminary studies of stable inhibition of miR-141 showed significant upregulation of the CSC marker CK5/6 mRNA and an expansion of CK5/6+ cells with progestin treatment. To investigate the role of miR-141 in hormone dependent breast cancer, we have measured levels of miR-141 and family in multiple luminal and basal breast cancer cell lines and in CK5/6+ and CK5/6- cells. We have also created cells with stable knockdown and overexpression of miR-141 to measure mRNA and protein levels of basal and luminal markers. We have validated potential miR-141 targets, including CK6, using luciferase reporter assays and miR-141 mimics to show direct targeting of miR-141 to their 3[apos]-UTRs. Finally, we have measured miR-141 levels in fractionated CSC and non-CSC populations and show that miR-141 specifically downregulated in the CSC fraction.[br]These data support a role for miR-141 in potentiating the transition from luminal cancer cells to CSCs. This is important as a specific role of miR-141 has not yet been described in the maintenance of differentiated phenotype, as it has for other family members. Defining the molecular mechanisms that promote breast CSCs will help design better strategies to prevent or target these cells in tumors.[br][br]Nothing to Disclose: JF-S, DMC, PH, PRP, DMC, BMJ, JKR, CAS 2012-06-23T12:15:00 342ABDE 2012-06-23T00:00:00 1899-12-30T12:15:00 1992 38 84 OR05-5 OR42-01 Saturday 29 2012


30 ENDO12L_OR05-6 ORAL SESSION: Hormone Dependent Tumors (11:15 AM-12:45 PM) Regulation of Type 1 CRH-R Splicing and MCF-7 Cell Responsiveness to CRH by Estrogens: Implications for Breast Cancer Tumor Biology Suchita Lal, Anna Allan, Dimitris Grammatopoulos University of Warwick, Coventry, UK Corticotropin-releasing hormone (CRH), the 41 amino acid hypothalamic peptide that plays a fundamental role in adaptation to stressful stimuli, is emerging as an important regulator of breast cancer cell biology. Previous studies supported a positive effect of CRH on tumour growth by showing increased proliferation, migration and actin polymerization in the 4T1 mouse mammary tumour cell line (1,2). This proposed tumor-promoting action of CRH has been challenged by other studies suggesting an anti-neoplastic potential of CRH through anti-proliferative and differentiation-inducing effects (3,4). Moreover, the use of corticorelin acetate (CrA), a synthetic CRH analog in the preclinical model of MX-1 breast carcinoma, demonstrated substantial antitumor activity, by exerting antiangiogenic effects (5). To understand the role of CRH in breast cancer biology, we used the oestrogen-responsive MCF-7 cell line and we identified a complex signalling network activated by type 1 CRH receptors (CRH-R1) involving activation of pro-survival and migration signalling molecules such as Akt, Hsp27 and p38 MAPK, and Focal Adhesion kinase (FAK) as well GSK3[beta]. Phosphorylation of the latter at Tyr216 was associated with a 50% decrease in [beta]-catenin protein levels, thus suggesting a negative effect on malignant cellular pathways. Interestingly, the signalling potency of CRH was reduced when cells were pre-treated with 10 nM 17[beta]-estradiol (E2). We investigated the profile of CRH-R1 mRNA variants and we identified significant increase by 4-fold of mRNA transcripts of the signaling impaired exon 12-missing variant CRH-R1d, in response to E2 treatment. This was associated with an increase in cytoplasmic CRH-R1 immunoreactivity, a characteristic of CRH-R1d protein expression. In silico studies identified the Serine/Arginine-rich splicing factor 6 (SRp55) as a potential regulator of exon 12 retention in the CRH-R1 mRNA transcript. Indeed, E2 treatment significantly attenuated SRp55 mRNA and protein expression in MCF-7 cells. Attenuation of SRp55 expression by either siRNA or E2-treatment led to a significant increase in CRH-R1d mRNA. These results suggest that CRH actions in MCF-7 cells are sensitive to estrogen-driven pathways that down-regulate expression of SRp55. This signaling modification drives CRHR1 gene splicing mechanisms towards generation of the signaling impaired CRH-R1d and would result in dampened responses to CRH.[br][br]1.Androulidaki A et al. Mol Cancer 2009; 8: 30. 2.Arranz A et al. Mol Cancer 2010; 9: 261. 3. Tjuvajev J et al In Vivo. 1998; 12:1-10. 4. Graziani G et al Mol Cell Endocrinol. 2007 264:44-94. 5. Gamez I et al. 2011; 67: 1415-22.[br][br]Sources of Research Support: The Pathological Society of Great Britain.[br][br]Nothing to Disclose: SL, AA, DG 2012-06-23T12:30:00 342ABDE 2012-06-23T00:00:00 1899-12-30T12:30:00 1271 38 85 OR05-6 OR42-01 Saturday 30 2012


31 ENDO12L_OR06-1 ORAL SESSION: Benign Thyroid Disease (11:15 AM-12:45 PM) Nutrient Restriction during Critical Illness Accelerates Recovery While Further Aggravating the Low T3 Syndrome: A Randomized Controlled Clinical Study Lies Langouche, Sarah Vander Perre, Mirna Bastos Marques, Anita Boelen, Michael Casaer, Greet Van den Berghe KU Leuven, Leuven, Belgium; Academic Medical Center, Amsterdam, Netherlands Critically ill patients reveal low circulating T3 and/or T4 levels in the absence of a rise in TSH. This constellation is labeled [apos]low T3 syndrome[apos] or [apos]nonthyroidal illness syndrome (NTI)[apos]. Interestingly, in healthy subjects, a low T3 syndrome is evoked by fasting in which it presumably mediates adaptation to reduced macronutrient availability. The contribution of restricted nutrition during human critical illness in bringing about the NTI currently remains unclear.[br]We recently performed a large randomized, controlled, multicenter trial, in which we compared early initiation of parenteral nutrition to supplement insufficient enteral feeding ([apos]early PN[apos] group) with tolerating pronounced nutritional deficit during the first week in ICU ([apos]late PN[apos] group). Tolerating the nutritional deficit resulted in faster recovery from organ failure and fewer complications, as compared with early supplementation with parenteral feeding (1). The aim of the current study was to investigate the impact of early nutritional deficit versus early feeding on the NTI of critical illness, in relation to outcome.[br]Of a subgroup of patients in this trial, identifiable upon ICU admission, for whom early enteral nutrition was surgically contraindicated and for whom the study actually compared early PN with full fasting for the first week in ICU, we randomly selected 280 patients for this analysis. In the [apos]early PN[apos] group (n=140), parenteral nutrition was initiated within 48 hours after ICU admission, whereas in the [apos]late PN[apos] group (N=140), parenteral nutrition was only initiated on day 8 when still in ICU. Serum isolated from blood samples collected on admission, on ICU day 3 and 5 and on the last day in ICU were quantified for circulating levels of TT3, TT4 and leptin using commercial assays.[br]The decrease over time in TT3 was significantly blunted by early PN, whereas the decrease in TT4 was only transiently affected. These changes coincided with a pronounced increase in circulating leptin over time, although the circulating levels of leptin did not correlate with circulating thyroid hormone levels. Analyses for circulating rT3 and TSH are ongoing and will be presented. Also, further analyses will assess the extent to which the changes in thyroid hormones, brought about by the nutritional management, explain the clinical benefits of nutrient restriction.[br][br](1) Casaer M et al., 2011 NEJM; 365:506.[br][br]Nothing to Disclose: LL, SVP, MBM, AB, MC, GVdB 2012-06-23T11:15:00 Theater B 2012-06-23T00:00:00 1899-12-30T11:15:00 663 39 86 OR06-1 OR39-01 Saturday 31 2012


32 ENDO12L_OR06-2 ORAL SESSION: Benign Thyroid Disease (11:15 AM-12:45 PM) Hypothyroidism in Critically Ill Patients: Outcomes and Effects of Thyroid Hormone Replacement Katayoun Edalat Parsi, Marianne Ghobrial, Ivance Pugoy, Vijay Balasubramanian, Soe Naing UCSF Fresno Center for Medical Education and Research, Fresno, CA; Fresno Medical Education Program, University of California, San Francisco, Fresno, CA; Fresno Medical Education Program, University of California, San Francisco, Fresno, CA BACKGROUND:[br]Hypothyroidism has been associated with increased mortality in critically ill patients. However, there is a paucity of literature evaluating impact of thyroid hormone replacement (THR) on the mortality rate (MR) in critically ill patients with hypothyroidism.[br]AIM: To evaluate the outcomes and effects of THR in critically ill patients with hypothyroidism in medical intensive care unit (MICU).[br]STUDY DESIGN:[br]This is a retrospective study of the patients admitted to the MICU at a community hospital from 2004 to 2010. Hypothyroidism was defined as admitting thyroid stimulating hormone (TSH) of [gt]30mU/L. The patients were divided into 3 groups based upon total levothyroxine (LT4) dose received during first week of MICU stay: [lt]500 mcg (low dose group or LD), 500-1000mcg (medium dose group or MD) and [gt]1000mcg (high dose group or HD).[br]RESULTS:[br]We studied a total of 36 patients (mean age 68.1years (SD[plusmn]17.7), men 47.2%, BMI 28.2 (SD[plusmn]9.6), mean TSH 92.5mU/L (SD[plusmn]95.4), Hispanics 50%, known primary hypothyroidism 92%, mean total LT4 dose received during the first week of ICU stay 846mcg (SD[plusmn]425), total hospital MR 31%). There were 8 patients in LD, 13 in MD and 15 in HD. None of the patients received liothyronine (T3). There was no statistically significant difference in mean age (68.8 vs 68.1 vs 67.7 years), gender (men 38 vs 62 vs 46%), mean BMI (29.7 vs 25.8 vs 29.6), mean TSH (105 vs 71 vs 104;NR 0.35-5.5 mU/L), mean FT4 (0.6 vs 0.6 vs 0.7;NR 0.89-1.76 ng/dl), mean APACHE II score (22.3 vs 19.1 vs 19.4), mean Glasgow Coma Scale (8.6 vs 12.7 vs 13.2), portions of the patients who received intravenous LT4 (25% vs 38% vs 40%) among 3 groups (LD vs MD vs HD). However the differences in mean total LT4 dose (272 vs 725 vs 1260 mcg) were statistically significant (P[lt]0.0001). Overall, total hospital MR was 50%, 38% and 13% in LD, MD and HD, respectively. MICU length of stay was 9.1, 9.2 and 15.6 days and total hospital stay was 15.8, 22.8 and 27.9 days in LD, MD and HD, respectively.[br]CONCLUSIONS:[br]1. Total hospital MR was 31% in critically ill patients with hypothyroidism who had admission TSH of [gt]30.[br]2. MR was inversely proportional to the total dose of THR received during the first week of MICU stay.[br]3. Aggressive THR during the first week of MICU stay in critically ill hypothyroid patients may improve survival.[br]Further studies with larger number of cases are needed to confirm the findings.[br][br]Nothing to Disclose: KEP, MG, IP, VB, SN 2012-06-23T11:30:00 Theater B 2012-06-23T00:00:00 1899-12-30T11:30:00 682 39 87 OR06-2 OR39-01 Saturday 32 2012


33 ENDO12L_OR06-3 ORAL SESSION: Benign Thyroid Disease (11:15 AM-12:45 PM) Cardiovascular Morbidity after Surgical Treatment of Hyperthyroidism [mdash] A Nationwide Cohort Study with a Long-Term Follow-Up Essi Ryodi, Jorma Salmi, Matti Valimaki, Anssi Auvinen, Pia Jaatinen, Heini Huhtala, Rauni Saaristo, Saara Metso Tampere University Hospital, Tampere, Finland; Tampere University Hospital, Tampere, Finland; Helsinki University Central Hospital, Helsinki, Finland; University of Tampere, Tampere, Finland; University of Tampere, Tampere, Finland; Tampere University Hospital, Tampere, Finland [bold]OBJECTIVE[/bold] Previous studies suggest that hyperthyroid patients remain at an increased risk of cardiovascular morbidity after restoring euthyroidism. The mechanisms of the increased risk and its dependency on different treatment modalities of hyperthyroidism remain unclear. The aim of this long-term follow-up study was to compare the cardiovascular causes of hospitalization in hyperthyroid patients treated with thyroidectomy with those of an age- and gender-matched reference population.[br][bold]PATIENTS AND MEASUREMENTS[/bold] A population-based cohort study was conducted among the 4334 hyperthyroid patients treated with thyroidectomy between years 1986 and 2007 in Finland and among 12,991 reference subjects. Information on hospitalizations was obtained from the nationwide Hospital Discharge Registry. New events were analyzed as the main outcome, including only the first hospitalization due to the given indication.[br][bold]RESULTS[/bold] The median follow-up time of the patients was 10.5 years. The hospitalization rate due to cardiovascular diseases was higher among the patients with hyperthyroidism than in the control population (240.5 vs 206.2 per 10 000 patient-years, rate ratio (RR) 1.17, 95% CI 1.09-1.26). The risk was sustained up to two decades after thyroidectomy. Hospitalizations due to arrhythmias (RR 1.50), hypertension (RR 1.27), and other cardiovascular diseases (RR 1.54) including non-bacterial endo-, peri- and myocardial diseases, valvular diseases, and cardiomyopathy were significantly more frequent among the patients than the controls. No significant difference was found between the patients and controls for coronary artery disease (RR 1.04), cerebrovascular diseases (RR 1.12) and diseases of other arteries and veins (RR 1.11) or heart failure (RR 1.18).[br][bold]CONCLUSIONS[/bold] In line with previous findings on patients treated with radioactive iodine, the current study shows that also hyperthyroid patients treated with thyroidectomy have an increased risk of hospitalization due to cardiovascular diseases. The risk is sustained up to two decades after surgery, which is regarded as an effective treatment modality for hyperthyroidism.[br][br]Sources of Research Support: This work was supported by grants from the Research Funding of the Pirkanmaa Hospital District.[br][br]Nothing to Disclose: ER, JS, MV, AA, PJ, HH, RS, SM 2012-06-23T11:45:00 Theater B 2012-06-23T00:00:00 1899-12-30T11:45:00 673 39 88 OR06-3 OR39-01 Saturday 33 2012


34 ENDO12L_OR06-4 ORAL SESSION: Benign Thyroid Disease (11:15 AM-12:45 PM) Age-Related Modifications of the Relationship between Thyroid Status and Lipoprotein Profile: Results from a Large Cross-Sectional Study Sara Tognini, Antonio Polini, Giuseppe Pasqualetti, Silvia Cottone, Silvia Ursino, Fabio Monzani University of Pisa, Pisa, Italy Background: The relationship between serum lipids and subclinical hypothyroidism is controversial. We evaluated the possible influence of age and gender in a large cohort of patients attending a thyroid outpatient clinic.[br]Patients [amp] Methods: 2308 consecutive patients (1874 F, mean age 47[plusmn]4.1 years) were submitted to complete clinical workup. Thyroid status and lipoprotein profile were assessed after an overnight fast. Study participants were divided into subgroups according to age (1st: 10-29, 2nd: 30-49, 3rd: 50-64, 4th: [gt]65 years) and serum TSH values (1st: [lt]0.36; 2nd: [gt]0.36 and [lt]3.6; 3rd: [gt]3.6 and [lt]10.0; 4th: [gt]10.0 mIU/l).[br]Results: Males and females were matched for age and blood pressure while body mass index was slightly higher in men (p=0.05). The prevalence of autoimmunity was significantly higher in women than men (55.4% Vs 39.9%, p=0.0002) but no gender difference in TSH frequency distribution was observed. Total (TC) and LDL cholesterol (LDLc) values (p[lt]0.0003 and p[lt]0.003, respectively) as well as LDL/HDLc ratio (p[lt]0.03) were significantly elevated in unselected women of the 4th TSH group ([gt]10 mIU/l) but, while analyzing women of different age groups, also those of the 3rd TSH group older than 50 years showed an atherogenic lipid profile (TC and LDLc p=0.01 and LDL/HDLc ratio p=0.02 Vs euthyroid women). Among unselected men only those of the 4th TSH group had significantly elevated triglyceride (TG) (p[lt]0.0001) but not cholesterol values. However, men of the 3rd and 4th TSH groups older than 65 years showed significantly higher TC, LDLc values and LDL/HDLc ratio too (p=0.03, p=0.02 and p=0.01, respectively Vs euthyroid men). In the final model of stepwise regression analysis, which explained almost 40% of serum TC and LDLc variations, age had the highest standardized coefficient (0.36 and 0.37, respectively), followed by TSH (0.20 and 0.11, respectively) and FT4 (-0.11 and [ndash]0.09, respectively).[br]Conclusions: The current study, while confirming a gender related differentiation in the relationship between thyroid function and lipid profile, firstly documents that the impact of mild thyroid failure on serum lipids is substantially influenced by age in both genders. Overall, our data delineate a definite interrelationship between thyroid function, age and gender and may, at least partially, explain the conflicting results of the literature regarding the impact of slightly elevated TSH value on circulating lipid parameters.[br][br]Nothing to Disclose: ST, AP, GP, SC, SU, FM 2012-06-23T12:00:00 Theater B 2012-06-23T00:00:00 1899-12-30T12:00:00 1075 39 89 OR06-4 OR39-01 Saturday 34 2012


35 ENDO12L_OR06-5 ORAL SESSION: Benign Thyroid Disease (11:15 AM-12:45 PM) Subclinical Thyroid Dysfunction Is Not Associated with Risk of Cognitive Decline or Dementia in Older Adults Eve Bloomgarden, Alice M Arnold, Michelle Carlson, Lewis Kuller, Oscar Lopez, Anne R Cappola University of Pennsylvania, Philadelphia, PA; University of Washington, Seattle, WA; Johns Hopkins University, Bloomberg School of Public Health, Baltimore, MD; University of Pittsburgh, Pittsburgh, PA; University of Pittsburgh, Pittsburgh, PA [bold]Background:[/bold] Subclinical thyroid dysfunction is common in older individuals and has been linked to neurocognitive impairment. We assessed the risks of cognitive decline and dementia in older adults with subclinical thyroid dysfunction.[br][bold]Methods:[/bold] Annual Modified Mini Mental Examination (3MSE), Digit Symbol Substitution Test (DSST) scores and incident dementia were analyzed from US community-dwelling men and women aged [ge]65 y in the Cardiovascular Health Study (CHS) who were not taking thyroid hormone preparations. Generalized estimating equations were used to estimate the difference in rate of change in 3MSE and DSST scores comparing subclinical hypothyroid (Shypo; n=624) and subclinical hyperthyroid (Shyper; n=74) groups to the euthyroid group (n=3894). Informative missingness was assessed in sensitivity analyses using weights defined by the inverse probability of dropping out. Cox proportional hazard models were used to examine the relationship between baseline thyroid status and incident dementia. All models were adjusted for age, sex, race, and education. Analyses were repeated in a subset of participants who had a second set of thyroid measurements to define persistent, transient, or progressive Shypo and Shyper.[br][bold]Results:[/bold] There was no difference in rate of change of 3MSE or DSST score by baseline or persistent thyroid category, although there was a suggestion of greater decline in DSST scores in the baseline Shyper group compared to the euthyroid group (-0.79 vs. -0.47; p=0.06), which was confirmed in the sensitivity analysis (-0.94 vs. -0.44; p=0.004). In the small group of Shypo participants who progressed to overt hypothyroidism, a greater decline in DSST was observed, approaching statistical significance (-1.96 vs. -0.85; p=0.07). There were 402 cases (339 euthyroid, 58 Shypo, 5 Shyper) of incident dementia over a mean follow-up of 5.4 years. We found no association of thyroid status with incident dementia. The hazard ratio was 1.06 (0.80- 1.40) for the Shypo group and 1.32 (0.54-3.19) for the Shyper group. Findings were similar in models of persistence.[br][bold]Conclusions: [/bold]We found no difference in cognitive decline or development of dementia in older individuals with baseline or persistent Shypo. There may be a greater decline in those who progress to overt hypothyroidism and in those with Shyper. We find no evidence for a clinically significant impact of subclinical thyroid dysfunction on cognitive function in this population.[br][br]Sources of Research Support: R01 AG032317 to ARC.[br][br]Nothing to Disclose: EB, AMA, MC, LK, OL, ARC 2012-06-23T12:15:00 Theater B 2012-06-23T00:00:00 1899-12-30T12:15:00 657 39 90 OR06-5 OR39-01 Saturday 35 2012


36 ENDO12L_OR06-6 ORAL SESSION: Benign Thyroid Disease (11:15 AM-12:45 PM) Women with High Early Pregnancy Urinary Iodine Levels Are at Risk of Hyperthyroid Newborns: The Generation R Study Marco Medici, Akhgar Ghassabian, Willy Visser, Sabine MPF de Muinck Keizer-Schrama, Herbert Hooijkaas, Jacoba J Bongers-Schokking, Henning Tiemeier, Alec Ross, Theo J Visser, Yolanda B de Rijke, Robin P Peeters Erasmus Medical Center, Rotterdam, Netherlands; Erasmus Medical Center [ndash] Sophia Children[apos]s Hospital, Rotterdam, Netherlands; Erasmus Medical Center [ndash] Sophia Children[apos]s Hospital, Rotterdam, Netherlands; Erasmus Medical Center [ndash] Sophia Children[apos]s Hospital, Rotterdam, Netherlands; Erasmus Medical Center, Rotterdam, Netherlands; Erasmus Medical Center, Rotterdam, Netherlands; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands Introduction:[br]Both iodine deficiency and iodine excess can lead to thyroid dysfunction. During pregnancy, iodine requirements increase and maternal iodine deficiency can result in developmental brain damage. Most studies on iodine and pregnancy focus on regions with (mild) iodine deficiency. As iodine intake is highly variable within populations, it is remarkable to note that little data on the effects of variation in iodine status on maternal and fetal thyroid function are available from iodine-sufficient populations.[br]Materials and methods:[br]Urinary iodine, serum TSH, FT4 and TPO-antibody (TPOAb) levels were determined in early pregnancy (13.3 (1.7) wk) in 1154 pregnant women from the Generation R Study, a population-based cohort study in Rotterdam, The Netherlands, which is considered an iodine-sufficient area. Cord serum TSH and FT4 levels were available in 1068 newborns. Informed consent was obtained from all participants.[br]Results:[br]The median urinary iodine level was 222.5 [mu]g/L (range: 9.3 [ndash] 1743.5 [mu]g/L), indicating an iodine-sufficient population (1). 30.8% and 11.5% had urinary iodine levels [lt] 150 and [gt] 500 [mu]g/L, respectively.[br]When comparing mothers with urinary iodine levels [lt] 150 vs [ge] 150 [mu]g/L, and [gt] 500 vs [le] 500 [mu]g/L, there were no differences in the risk of maternal hypothyroxinemia, (subclinical) hypo- or hyperthyroidism. However, mothers with urinary iodine levels [gt] 500 [mu]g/L had a 2.7 times increased risk (5.6[plusmn]1.4 vs 2.1[plusmn]0.5 %, [italic]P[/italic] = 0.035) of a newborn with suppressed cord TSH levels (TSH [lt] 3.10 mU/L ([lt] 2.5[sup]th[/sup] percentile)), as well as a 5.2 times increased risk (3.1[plusmn]0.9 vs 0.6[plusmn]0.3 %, [italic]P[/italic] = 0.016) of a hyperthyroid newborn (TSH [lt] 3.10 mU/L ([lt] 2.5[sup]th[/sup] percentile) and FT4 [gt] 30.1 pmol/L ([gt] 97.5[sup]th[/sup] percentile)). These mothers had newborns with higher cord FT4 levels (21.7[plusmn]0.3 vs 21.0[plusmn]0.1 pmol/L, [italic]P[/italic] = 0.036).[br]Maternal urinary iodine levels [lt] 150 [mu]g/L were not associated with newborn thyroid dysfunction.[br]Conclusion:[br]In an iodine-sufficient population, women with higher urinary iodine levels have a substantial increased risk of a hyperthyroid newborn. In contrast, women with lowest urinary iodine levels did not have a higher TSH. These data provide insight into the effects of variation in maternal iodine status on maternal and fetal thyroid dysfunction.[br][br](1) World Health Organization United Nations Children[apos]s Fund, International Council for the Control of Iodine Deficiency Disorders. 2007 Assessment of Iodine Deficiency Disorders and Monitoring Their Elimination. 3rd ed. Geneva: WHO.[br][br]Nothing to Disclose: MM, AG, WV, SMPFdMK-S, HH, JJB-S, HT, AR, TJV, YBdR, RPP 2012-06-23T12:30:00 Theater B 2012-06-23T00:00:00 1899-12-30T12:30:00 826 39 91 OR06-6 OR39-01 Saturday 36 2012


1 ENDO12L_OR01-1 ORAL SESSION: Endocrine Gene Regulatory Circuits (11:15:00 AM-12:45:00 PM) A Pitx2-miRNA-200c-Noggin Regulatory Axis Regulates BMP Signaling and Anterior Pituitary Cell Type Specification Huojun Cao, Xiao Li, Jianbo Wang, Zhao Sun, Zichao Zhang, Brad A Amendt Texas A[amp]M Health Science Center, Institute of Biosciences and Technology, Houston, TX The microRNA-200 family plays a critical role in cancer cell progression and regulation of the epithelial-to-mesenchymal transition during embryonic development and cancer. We previously reported a role for mature miRNAs in pituitary development, hormone levels and gene expression. However, the specific miRNAs that regulate pituitary cell types remains unknown. The miRNA-200 family is specifically up-regulated during pituitary development and in gondadotropes, somatotropes and lactotropes, but not in corticotropes. The miRNA-200 family expression increases with differentiation. BMP signaling is required for invagination of Rathke[apos]s pouch and over-expression of Noggin (a BMP2/4 antagonist) inhibits pituitary cell lineage differentiation. BMP4 signaling and therefore repressed Noggin expression is required for the continued progression of pituitary development. We demonstrate that miRNA-200c targets Noggin and allows for BMP signaling to regulate gene expression and differentiation of pituitary lineage cells. Interestingly, miRNA-200c and 200b expression are increased in PITX2C over expression mouse epithelial tissues. Chromatin immunoprecipiation assays reveal endogenous PITX2 binding to the miRNA-200c-141 promoter. Transfection of PITX2 in cells activates endogenous miRNA-200c and the miRNA-200c enhancer reporter. The Pitx2 transcription factor regulates early pituitary developmental programs and we demonstrate Pitx2 regulates the miRNA-200 family during development. The activation of miRNA-200c indirectly regulates BMP signaling by targeting and repressing Noggin expression. This process allows for pituitary progenitor cells to become differentiated cells of the anterior pituitary. These studies reveal a multistep transcriptional program involving a Pitx2-miRNA200-Noggin axis during pituitary cell differentiation.[br][br]Sources of Research Support: National Institute of Dental and Craniofacial Research grants DE13941 and DE018885 awarded to BAA.[br][br]Nothing to Disclose: HC, XL, JW, ZS, ZZ, BAA 2012-06-23T11:15:00 320 2012-06-23T00:00:00 1899-12-30T11:15:00 2137 34 56 OR01-1 OR16-01 Saturday 1 2012


2 ENDO12L_OR01-2 ORAL SESSION: Endocrine Gene Regulatory Circuits (11:15:00 AM-12:45:00 PM) Evidence That microRNA-29 Downregulates Multiple Growth-Promoting Genes and Thus Contributes to Juvenile Growth Deceleration Fariha Kamran, Anenisia C Andrade, Samuel Clokie, Geoffrey A Rezvani, Ola Nilsson, Jeffrey Baron, Julian C Lui National Institute of Child Health and Human Development/National Institutes of Health, Bethesda, MD; Karolinska Institutet and Karolinska University Hospital, Stockholm, Sweden; St Christopher[apos]s Hospital for Children, Philadelphia, PA Infancy is characterized by rapid body growth, which slows with increasing age and eventually ceases due to a progressive decline in cell proliferation that occurs simultaneously in multiple organs. We previously showed evidence that this decline in proliferation is driven in part by postnatal downregulation of a large set of growth-promoting genes in multiple organs (Endocrinology 150:1791-800, 2009; FASEB J 24:3083-92, 2010). We hypothesized that this growth-limiting genetic program is orchestrated by microRNAs, which are short non-coding RNAs that bind to target sequences in multiple mRNAs to inhibit translation or stimulate mRNA degradation. A bioinformatic algorithm (Partek Genomics Suite 6.6) was used to identify microRNA target sequences in genes that are commonly downregulated in mouse kidney and lung as growth slows from 1 to 4 weeks of age. The analysis predicted that the downregulated genes are strongly enriched with target sequences for microRNA-29 family members (miR-29a, b, and c, P [lt] 0.00001 each). Concomitantly, we analyzed changes in microRNA expression in juvenile mice as growth slows with age, from 1 to 6 weeks by microarray (Agilent Mouse Genome VI). miR-29a, b, and c were strongly upregulated with age in both kidney and lung (all [gt]8-fold increase, false discovery rate [lt] 0.05). Real-time PCR confirmed significant (P [lt]0.05) upregulation with age, from 1 to 4 weeks, of miR-29a (lung, 15-fold; kidney, 27-fold), miR-29b (lung, 9-fold; kidney, 32-fold) and miR 29c (lung, 16-fold; kidney, 58-fold). We next focused on 3 age-downregulated genes that are predicted targets of miR-29: Igf-1, Igf2bp1, and Mest. The 3[apos]-UTR of each gene was placed downstream of a luciferase reporter and transfected into HEK293 cells. Co-transfection with miR-29a, b or c mimics suppressed luciferase activity, confirming that the mRNAs are actual targets of miR-29. In summary: (i) miR-29a, b and c are strongly upregulated with age in multiple organs during juvenile life, (ii) miR-29 target sequences are strongly overrepresented in genes that are downregulated with age in multiple organs and (iii) some of these predicted targets have been experimentally confirmed. Taken together, the findings suggest that upregulation of miR-29a, b, and c during juvenile life downregulates multiple growth-promoting genes, thus contributing to physiological slowing and eventual cessation of body growth.[br][br]Nothing to Disclose: FK, ACA, SC, GAR, ON, JB, JCL 2012-06-23T11:30:00 320 2012-06-23T00:00:00 1899-12-30T11:30:00 1392 34 57 OR01-2 OR16-01 Saturday 2 2012


3 ENDO12L_OR01-3 ORAL SESSION: Endocrine Gene Regulatory Circuits (11:15:00 AM-12:45:00 PM) [italic]ARTD1[/italic] Deletion Causes Increased Hepatic Lipid Accumulation in Mice on High-Fat Diet and Impairs Adipocyte Function and Differentiation Suheda Erener, Ali Mirsaidi, Mareike Hesse, Andre N Tiaden, Helga Ellingsgaard, Radina Kostadinova, Marc Y Donath, Peter J Richards, Michael O Hottiger University of Zurich, Zurich, Switzerland; University of Zurich, Zurich, Switzerland; University Hospital Basel, Basel, Switzerland ADP-ribosyltransferase Diphtheria toxin-like 1 (ARTD1) is a chromatin-associated enzyme involved in regulating metabolic homeostasis. We provided earlier evidence that ARTD1-dependent ADP-ribose polymer (PAR) formation increases during adipocyte development and, at late time points of adipogenesis, is involved in the sustained expression of [italic]PPAR[gamma]2[/italic] and of PPAR[gamma]2 target genes. During adipogenesis, ARTD1 is recruited to [italic]PPAR[gamma]2[/italic] target genes such as [italic]CD36[/italic] or [italic]aP2[/italic] in a PAR-dependent manner. Interestingly, activation of ARTD1 enzymatic activity is prevented with a Toposiomerase II inhibitor (1).[br]The liver is at the core of glucose and lipid metabolism and significantly affected by obesity and the metabolic syndrome. Here, we will show that on high fat diet (HFD), mice lacking [italic]ARTD1[/italic] developed exacerbated hepatic steatosis. [italic]ARTD1[/italic][sup]-/-[/sup] mice had a 19% higher liver weight than wildtype (WT) animals and also exhibited an increased serum concentration of free fatty acids (42% increase) and cholesterol (38% increase) and impaired glucose tolerance. In addition, adipocyte function and size were significantly reduced in [italic]ARTD1 [/italic][sup]-/-[/sup] mice on HFD (7794 [mu]m[sup]2[/sup] for WT and 5579 [mu]m[sup]2[/sup] for [italic]ARTD1[/italic][sup]-/-[/sup] mice). The significantly reduced adipogenic differentiation of adipose-derived stromal cells (ASCs) isolated from [italic]ARTD1 [/italic][sup]-/-[/sup] mice (28% vs. 11% Oil red O positive cells in WT and [italic]ARTD1 [/italic][sup]-/-[/sup] ASCs, respectively) suggested adipogenesis as the underlying cause for this adipose tissue malfunction. This function of ARTD1 was specific for adipogenesis since osteogenic differentiation was not affected by the [italic]ARTD1[/italic] deletion. Our data suggest a novel role for ARTD1 in the development of hepatic steatosis as a consequence of impaired adipogenesis and adipocyte malfunction, which can subsequently lead to non-alcoholic fatty liver disease.[br][br](1) Erener S et al., Mol Endocrinol. 2012; 26(1):79-86.[br][br]Sources of Research Support: This work was supported in part by the University Research Priority Program [ldquo]Integrative Human Physiology[rdquo] at the University of Zurich, the Swiss National Science Foundation Grant 31-122421 and 31-138667 and the Kanton of Zurich (to M.O.H.).[br][br]Nothing to Disclose: SE, AM, MH, ANT, HE, RK, MYD, PJR, MOH 2012-06-23T11:45:00 320 2012-06-23T00:00:00 1899-12-30T11:45:00 1436 34 58 OR01-3 OR16-01 Saturday 3 2012


4 ENDO12L_OR01-4 ORAL SESSION: Endocrine Gene Regulatory Circuits (11:15:00 AM-12:45:00 PM) The miR-200 Family and ZEB Transcription Factors Serve Important Roles in Developmental Induction of Surfactant Protein-A (SP-A) Gene Expression in Human Fetal Lung Houda Benlhabib, Carole R Mendelson University of Texas Southwestern Medical Center, Dallas, TX Differentiation of the fetal lung and development of the capacity to synthesize pulmonary surfactant are essential for transition of the fetus from an aqueous to an air-breathing environment. In recent years, considerable progress has been made in our understanding of the transcriptional mechanisms for differentiation of the developing lung and expression of genes encoding the surfactant proteins. To further define mechanisms for type II cell differentiation and developmental induction of surfactant synthesis, we are investigating the potential role of microRNAs (miRNAs, miRs). miRNAs are evolutionarily conserved [sim]22 nucleotide, single-stranded noncoding RNAs that, inhibit gene expression by binding to the 3[apos]-untranslated regions of their target mRNAs through base-pairing, resulting in inhibition of mRNA translation and/or increased mRNA degradation. Previously, we observed that midgestation human fetal lung explants differentiate spontaneously in organ culture in serum-free medium. The rate of type II cell differentiation and expression of the gene encoding the major surfactant protein, SP-A, are markedly induced by cAMP. To identify and characterize differentially regulated miRNAs in mid-gestation human fetal lung explants upon type II pneumocyte differentiation in culture, we performed miRNA microarray analysis of RNA isolated from epithelial cells from human fetal lung explants before and after 48 and 96 h of culture [plusmn] Bt[sub]2[/sub]cAMP. Interestingly, the miR-200 family was found to be significantly upregulated with type II cell differentiation. Moreover, the induction of miR-200 family members was inversely correlated with expression of their known targets, transcription factors ZEB1 and ZEB2. Interestingly, overexpression of ZEB1 or ZEB2 in cultured type II cells using recombinant adenoviruses blocked cAMP stimulation of SP-A expression. On the other hand, cAMP, which promotes type II cell differentiation, caused a marked inhibition of ZEB expression. Notably, culture of human fetal lung epithelial cells with active TGF-[beta]1, known to induce epithelial to mesenchymal transition in epithelial cells and to inhibit type II cell differentiation, induced expression of ZEBs and inhibited SP-A expression. These findings suggest that the miR-200 family and ZEB1/2, which exist in a double-negative feedback loop, serve important regulatory roles in the developmental regulation of type II cell differentiation and SP-A expression in human fetal lung.[br][br]Sources of Research Support: NIH R01 HL050022.[br][br]Nothing to Disclose: HB, CRM 2012-06-23T12:00:00 320 2012-06-23T00:00:00 1899-12-30T12:00:00 1931 34 59 OR01-4 OR16-01 Saturday 4 2012


5 ENDO12L_OR01-5 ORAL SESSION: Endocrine Gene Regulatory Circuits (11:15:00 AM-12:45:00 PM) Influence of Prenatal Steroid Exposure on Insulin Sensitivity and DNA Methylation in Mice Tiffany Sue Musick, Scott A Sands, Jill D Jacobson Children[apos]s Mercy Hospitals and Clinics, Kansas City, MO; Univeristy of Missouri-Kansas City School of Medicine, Kansas City, MO Objective: Stimulatory G proteins play a key role in energy metabolism. Loss of function mutations in the [italic]Gnas[/italic] gene, encoding the guanine nucleotide alpha subunit (G[alpha]s), lead to obesity and insulin resistance. Maternally inherited mutations of the [italic]Gnas[/italic] gene are known to cause obesity in both mice and humans. We have recently found that mice bearing inactivating mutations in the [italic]Gnas [/italic]gene display low estradiol levels during pregnancy. We hypothesized that alterations in the prenatal hormonal milieu would affect later insulin sensitivity, G protein expression, and/or epigenetic phenomenon, specifically DNA methylation.[br]Methods: All experiments were approved by the UMKC Animal Care and Use Committee. Pregnant female mice were treated with 17 [beta]-estradiol (.1 [micro]g/day) on days 16-18 or 5-21, dihydrotestosterone (DHT) (250 [micro]g/day) on days 16-18, or with vehicle consisting of charcoal-stripped, delipidated corn oil on days 5-21. At 6 months of age, we performed intraperitoneal glucose tolerance tests (ipgtt) and obtained insulin and leptin measurements in female mice. DNA, RNA, and protein were purified from hypothalamus and white adipose tissue (WAT). We performed quantitative PCR for G[alpha]s and G[alpha]q mRNA. Global DNA methylation was quantitated by ELISA.[br]Results: Prenatal exposure to estradiol led to a significant decrease in serum insulin levels at the peak time point (60 minute) in the ipgtt test[(mean [plusmn] SEM) 0.1796 [plusmn] 0.0269 vs. 0.4978 [plusmn] 0.0826]. In contrast, prenatal exposure to DHT resulted in significant increases in insulin levels at the same time point (0.8046 [plusmn] 0.1725 vs. 0.4978 [plusmn] 0.0826). Prenatal exposure to estradiol led to increased expression of G[alpha]s mRNA in the hypothalamus (166.4 [plusmn] 14.9 vs. 117.8 [plusmn] 14.2) and WAT (155.3 [plusmn] 15.7 vs. 54 [plusmn] 14.1). G protein mRNA expression in the hypothalamus correlated inversely with log 60 minute glucose measurements (rho = -0.62; p[lt]0.001). Prenatal exposure to estradiol lead to an increase in G[alpha]s mRNA expression in parallel with a decrease in global DNA methylation in the hypothalamus (0.87 [plusmn] 0.07 vs. 1.23 [plusmn] 0.11 % control).[br]Conclusions: Our study may provide insight into possible mechanisms for the effects of sex steroids on later development of insulin resistance. Prenatal exposure to estradiol increased insulin sensitivity and G protein expression in hypothalamus and WAT. Aberrant levels of estrogens or androgens may contribute to altered expression of G[alpha]s and insulin sensitivity in adipose cells.[br][br]Sources of Research Support: Endocrine Fellows Foundation Marilyn Fishman Grant for Diabetes Research Fall 2010.[br][br]Nothing to Disclose: TSM, SAS, JDJ 2012-06-23T12:15:00 320 2012-06-23T00:00:00 1899-12-30T12:15:00 725 34 60 OR01-5 OR16-01 Saturday 5 2012


6 ENDO12L_OR01-6 ORAL SESSION: Endocrine Gene Regulatory Circuits (11:15:00 AM-12:45:00 PM) T-Box 15 Is a Molecular Determinant of Fiber Type Distribution in Skeletal Muscle Kevin Y Lee, Cecile Vernochet, Stephane Gesta, C Ronald Kahn Joslin Diabetes Center, Boston, MA Tbx15 is a member of the T-box gene family that has been shown to have critical roles in mesoderm development and regulating mitochondrial mass and activity. We have previously shown that Tbx15 expression is increased in subcutaneous fat and decreased in the visceral fat of obese humans, and that Tbx15 overexpression in adipocytes in vitro impairs mitochondrial oxidation.[br]Skeletal muscle plays an important role in maintaining normal whole-body glucose metabolism and energy homeostasis, in part, due to its large mass and intrinsic oxidative capacity. In mammals, skeletal muscles are composed of a mosaic of different fiber types, and fiber type distribution correlates with glucose uptake and insulin resistance. Red oxidative fibers (Type I and IIa) are suited for endurance and use oxidative metabolism to generate ATP whereas white glycolytic fibers (Type IIx and IIb) are suitable for bursts of activity and rely on glycolytic metabolism.[br]In the present study, we demonstrate that Tbx15 is most highly expressed in skeletal muscle compared to other tissues. Interestingly, Tbx15 expression is increased in the skeletal muscle of obese humans. Using a Lac-Z reporter construct [italic]in vivo[/italic], we find that Tbx15 is almost exclusively expressed in Type IIb glycolytic muscle fibers. Global knockout of Tbx15 results in severe musculoskeletal developmental malformations. Heterozygous Tbx15 knockout (Tbx15[sup]+/-[/sup]) mice are developmentally normal in appear, but histological examination of the quadriceps from 6 week old animals revealed a 2.5-fold increase in the density of oxidative Type IIa fibers compared to controls and a corresponding decrease in Type IIb fibers. Furthermore, muscle of Tbx15[sup]+/-[/sup] animals displayed a constitutive activation of adenosine monophosphate kinase (AMPK) signaling. Muscles of Tbx15[sup]+/-[/sup] animals also exhibited a decrease in expression of insulin-like growth factor 2 (IGF-2) during embryogenesis. C2C12 myoblast cell lines with a stable knockdown of Tbx15 also exhibited a robust increase in AMPK signaling, and a 70% decrease in the expression of IGF-2. On the other hand, stable over-expression of Tbx15 in C2C12 myoblasts leads to a seven-fold increase in IGF-2 expression.[br]Taken together, these data demonstrate that Tbx15 controls AMPK signal, IGF-2 expression, and fiber type in skeletal muscle. Thus, changes in Tbx15 expression in muscle contributes to regulation of energy metabolism and muscle function.[br][br]Nothing to Disclose: KYL, CV, SG, CRK 2012-06-23T12:30:00 320 2012-06-23T00:00:00 1899-12-30T12:30:00 1635 34 61 OR01-6 OR16-01 Saturday 6 2012


7 ENDO12L_OR02-1 ORAL SESSION: Development [amp] Modulation of Hypothalamic Neuropeptidergic Neurons (11:15 AM-12:45 PM) Speed of GnRH Neuronal Migration Is Controlled by Leading Process Actin and Microtubule Flow Bruce Ian Hutchins, Ulrike Klenke, Susan Wray National Institutes of Health, Bethesda, MD The GnRH system is essential for proper function of the HPG axis. Correct placement of migrating GnRH neurons and their projections within the CNS is essential for reproductive function and occurs early in development. Failure to establish an adult-like distribution can lead to reproductive disorders such as Kallman[apos]s syndrome. Migration from proliferative zones to the proper location in the brain is a challenging task for many neurons. This is especially true for GnRH neurons, which must first enter the brain from the nasal region before beginning their journey through the forebrain. Originating in the nasal placode, GnRH neurons follow olfactory axons into the brain, while extracellular signals regulate movement of GnRH neurons along this pathway. A major unresolved question in the field is how these signals effect the mechanical changes that underlie cellular movement. We hypothesized that dynamic actin/microtubule interactions form an intracellular engine that pulls GnRH neurons forward along their migratory pathway in response to signaling pathways initiated by extracellular cues. To investigate this hypothesis, migrating GnRH neurons were imaged in mouse nasal explants, a powerful in vitro model that recapitulates this stage of embryonic development. GFP-tagged actin was imaged in the leading process of GnRH neurons. Actin flow toward the distal leading process correlated with movement of the cell body. This actin flow could be evoked by pharmacologically stimulating calcium release from stores via phospholipase C, while blocking calcium release through IP3 receptors slowed actin flow and reduced movement of the cell body. A similar behavior was found after examination of the leading process microtubules. Based on these results, we hypothesized that actin flow draws the nucleus up the leading process by pulling forward a [apos]microtubule cage[apos] surrounding the nucleus. Depolymerizing microtubules was found to reduce migration rates, consistent with this hypothesis. Experiments are currently testing whether microtubule movement requires actin dynamics. These results, when taken together, integrate information from signaling pathways to the molecular engines pulling the cell body forward to reveal the precise mechanisms used by cues in the environment to set the speed of migrating GnRH neurons.[br][br]Sources of Research Support: The authors are supported by the Intramural Research Program of the National Institute of Neurological Disorders and Stroke, National Institutes of Health (B.I.H., U.K. and S.W.) and a fellowship from the Pharmacology Research Associate (PRAT) Prorgam, National Institute of General Medical Sciences, National Institutes of Health (B.I.H.).[br][br]Nothing to Disclose: BIH, UK, SW 2012-06-23T11:15:00 371 2012-06-23T00:00:00 1899-12-30T11:15:00 827 35 62 OR02-1 OR20-01 Saturday 7 2012


8 ENDO12L_OR02-2 ORAL SESSION: Development [amp] Modulation of Hypothalamic Neuropeptidergic Neurons (11:15 AM-12:45 PM) Alterations in Hypothalamic Gene Expression of Reproductive Neuropeptides during the Pubertal Transition in Mice Sheila J Semaan, Alexander S Kauffman University of California-San Diego, La Jolla, CA Puberty onset is induced by the secretion of gonadotropin releasing hormone (GnRH). However, the mechanisms controlling and timing puberty onset remain poorly-defined, especially regarding the role of neuropeptides that act upstream of GnRH neurons. In mammals, kisspeptin ([italic]Kiss1[/italic] gene) and neurokinin B (NKB; [italic]Tac2[/italic] gene), stimulate GnRH secretion, while RFRP-3 (the mammalian homologue of GnIH; [italic]Rfrp[/italic] gene) inhibits the GnRH axis. In the rodent hypothalamus, [italic]Kiss1[/italic] is expressed in the anteroventral periventricular nucleus (AVPV) and the arcuate (ARC) nucleus, with neurons in the ARC also co-expressing [italic]Tac2[/italic]. RFRP-3 neurons reside exclusively in the hypothalamic dorsomedial nucleus (DMN). Previous studies reported that [italic]Kiss1[/italic] expression in the brain is higher in adults than prepubertal animals, but the exact peripubertal age(s) when [italic]Kiss1[/italic] first increases is unknown, as is the precise developmental pattern of [italic]Kiss1[/italic] expression specifically in the ARC and AVPV during the pubertal transition. Likewise, the specific developmental pattern of [italic]Tac2[/italic] and [italic]Rfrp[/italic] expression just before and during puberty is not well-characterized. Lastly, the pattern of developmental changes in each of these reproductive neuropeptide systems in relation to one another during puberty has not been determined. Using [italic]in situ[/italic] hybridization, we determined the detailed pubertal developmental profile (from PND 15 to PND 30) of [italic]Kiss1, Tac2[/italic], and [italic]Rfrp[/italic] gene expression in the AVPV, ARC, and DMN, as well as [italic]cfos[/italic] induction in each of these regions, in female mice. The mean age of vaginal opening was postnatal day (PND) 27.5. [italic]Kiss1[/italic] expression in the AVPV of females increased steadily over the pubertal transition, reaching adult levels around PND 28. The largest daily increase was noted on PND 24, several days before the mean age of vaginal opening. In the ARC, however, Kiss1 and Tac2 cell numbers remained fairly steady over the pubertal transition, showing only a very minor increase. [italic]Kiss1[/italic] neurons were not highly activated in the AVPV at any pubertal age, as measured by cfos coexpression. Interestingly, [italic]Rfrp[/italic] expression decreased significantly around PND 21, which may relate to RFRP-3[apos]s possible role as an inhibitor of the GnRH axis prior to puberty. Additional data from current RFRP-3- and Tac2-cfos coexpression assays will provide a more detailed picture of how these key reproductive neuropeptide systems change during the pubertal transition.[br][br]Sources of Research Support: R00 HD056157, R01 HD065856, U54 HD012303, F32 HD066849, and DERC P[amp]F grant DK063491.[br][br]Nothing to Disclose: SJS, ASK 2012-06-23T11:30:00 371 2012-06-23T00:00:00 1899-12-30T11:30:00 532 35 63 OR02-2 OR20-01 Saturday 8 2012


9 ENDO12L_OR02-3 ORAL SESSION: Development [amp] Modulation of Hypothalamic Neuropeptidergic Neurons (11:15 AM-12:45 PM) Intrauterine Vitamin D[sub]3[/sub] Deficiency Disrupts the Female Reproductive Axis Joseph B Davis, Marlena Petti, Thalia Segal, Genevieve Neal-Perry Montefiore Medical Center, Bronx, NY; Albert Einstein College of Medicine, Bronx, NY; North Shore Long Island Jewish Medical Center, Hyde Park, NY Vitamin D3[sub] [/sub](VD3) receptors are localized in the hypothalamic-pituitary-ovarian axis (HPO) suggesting VD3 may be important in reproductive physiology. Consistent with this hypothesisVD3 deficiency is more prevalent in women with reproductive dysfunction. VD3 insufficiency and deficiency has reached near epidemic levels in reproductive aged women, thus raising the possibility that intrauterine and lactational (developmental) VD3 deficiency adversely affects the reproductive physiology of exposed female offspring. [bold]Objective[/bold]: To determine if developmental VD3 deficiency affects puberty or HPO function in mice. [bold]Method[/bold]: Females (n=14-24) born to dams fed a sufficient (control) or deficient VD3 diet during pregnancy and lactation were weaned onto aVD3 sufficient diet. The effect of developmental VD3 deficiency on puberty (vaginal opening and 1st estrus), estrous cycling, ovarian physiology, glucose tolerance, GnRH neuronal density and pituitary responsiveness to GnRH peptide was determined. Data was analyzed with 2-way ANOVA or non-parametric tests. [bold]Results[/bold]: Young adult mice exposed to developmental VD3 deficiency had a normal pubertal transition. However developmental VD3 deficiency resulted in extended estrous cycles (p[lt]0.01) characterized by a prolonged diestrus (p[lt]0.05) and oligoovulation (p[lt]0.05). Developmental VD3 deficiency was associated arrested ovarian follicular development characterized by more early stage primordial (p[lt]0.01) and primary (p[lt]0.01) follicles than controls. Developmental VD3 deficiency was not associated with glucose intolerance. Developmental VD3 deficiency did not affect E2 negative feedback effects on pituitary gonadotropin (LH and FSH) release or did not affect the number of GnRH neurons in the hypothalamus of adult females. However, developmental VD3 deficiency was associated with reduced pituitary FSH (P[lt]0.01) and tended to have less LH release following a GnRH peptide challenge under E2 positive feedback conditions.[bold]Conclusion[/bold]: These data suggest that developmental VD3 deficiency reduces pituitary responsiveness and gonadotropin release in response to GnRH peptide. This most likely disrupts ovarian physiology (arrested follicular development) and results in prolonged estrous cycles characterized by oligoovulation. These preliminary data suggest that maternal VD3 deficiency may adversely affects the reproductive phenotype of exposed adult female offspring through effects on the pituitary function and secondary effects on the ovary.[br][br]Sources of Research Support: HD066355.[br][br]Nothing to Disclose: JBD, MP, TS, GN-P 2012-06-23T11:45:00 371 2012-06-23T00:00:00 1899-12-30T11:45:00 918 35 64 OR02-3 OR20-01 Saturday 9 2012


10 ENDO12L_OR02-4 ORAL SESSION: Development [amp] Modulation of Hypothalamic Neuropeptidergic Neurons (11:15 AM-12:45 PM) Leptin Is Not Required for Fertility Oulu Wang, Satoru Sakihara, Kolbein Gudmundsson, Joseph Majzoub Children[apos]s Hospital, Boston, MA Stress responses to acute stressors improve the chances of immediate survival, even at the expense of immediate reproductive fitness. Stress-induced elevation of glucocorticoids is associated with inhibition of the hypothalamic-pituitary-gonadal reproductive axis. Leptin-deficient ob/ob mice are infertile and obese, and leptin is considered necessary for fertility. However, ob/ob mice also have significantly elevated glucocorticoid levels, and these elevated glucocorticoid levels may be responsible for infertility in leptin-deficient animals. Additionally, glucocorticoids stimulate appetite, and two common side effects of glucocorticoid therapy are hyperphagia and weight gain. We hypothesized that reducing glucocorticoid levels in leptin-deficient animals would restore fertility and ameliorate obesity.[br]To study this putative role of corticosterone in leptin deficiency, we generated mice with genetic deficits in the synthesis of glucocorticoids by deleting corticotropin-releasing hormone (CRH), leptin, or both. CRH- and leptin-deficient mice had hypotrophic adrenal glands, low corticosterone, and undetectable leptin levels. Compared to ob/ob mice, these males had higher testosterone levels, larger testes, and normal testicular histology. Females had higher luteinizing hormone and estradiol levels, larger ovaries, and larger uteri. Both males and females successfully generated litters, despite having global leptin deficiency, demonstrating that leptin is not required for reproduction. Leptin- and corticosterone-deficient mice had reduced hyperphagia and [gt]50% weight loss compared to ob/ob animals. Body fat composition, plasma triglycerides, and liver lipid content were all normalized.[br]To address whether the restoration of fertility and amelioration of body weight were the result of changes in glucocorticoids or CRH, we treated mice with exogenous corticosterone. Mice deficient for both CRH and leptin, but not either alone, were rendered infertile by glucocorticoid supplementation, without any induced elevation in CRH. Glucocorticoid supplementation caused increased body weight and plasma triglyceride levels in the CRH- and leptin-deficient group only, suggesting that glucocorticoids modulate fertility and appetite under conditions of insufficient leptin. These data indicate that the removal of glucocorticoid excess in the setting of low leptin is sufficient to restore fertility and ameliorate obesity.[br][br]Sources of Research Support: DoD NDSEG; NSF GRFP; NIH.[br][br]Nothing to Disclose: OW, SS, KG, JM 2012-06-23T12:00:00 371 2012-06-23T00:00:00 1899-12-30T12:00:00 1920 35 65 OR02-4 OR20-01 Saturday 10 2012


11 ENDO12L_OR02-5 ORAL SESSION: Development [amp] Modulation of Hypothalamic Neuropeptidergic Neurons (11:15 AM-12:45 PM) Deletion of Dopamine D2 Receptors in Pituitary Lactotropes Increases Food Intake, Body Weight and Adiposity Guillermina Maria Luque, Maria Ines Perez Millan, Damasia Becu-Villalobos, Marcelo Rubinstein IBYME-CONICET, Buenos Aires, Argentina; INGEBI-CONICET, Buenos Aires, Argentina Prolactin secretion is tonically inhibited by dopamine acting on lactotrope dopamine D2 receptors (D2Rs). Consequently, female mice lacking D2Rs ([italic]Drd2[sup]-/-[/sup][/italic]) exhibit chronic hyperprolactinemia and pituitary lactotrope hyperplasia. Although hyperprolactinemia has been associated with increased food intake and body weight, [italic]Drd2[sup]-/-[/sup][/italic] female mice display normal body weight probably because of multiple overlapping phenotypes derived from the absence of D2Rs from all brain areas and peripheral tissues. To dissect the importance of lactotrope D2Rs in food intake and body weight regulation we generated mutant mice carrying targeted [italic]Drd2 [/italic]deletions specifically in lactotropes (lac[italic]Drd2[sup]-/-[/sup][/italic]) using Cre/[italic]LoxP[/italic] technology. As expected, lac[italic]Drd2[sup]-/-[/sup][/italic] females are hyperprolactinemic whereas neuronal and peripheral D2Rs appear to function normally as determined by haloperidol-induced catalepsia and insulin serum levels. In addition, lac[italic]Drd2[sup]-/-[/sup][/italic] females showed increased body weight gain and adiposity in comparison with age-matched [italic]Drd2[sup]loxP/loxP[/sup][/italic] and [italic]Drd2[sup]-/-[/sup][/italic] female mice. Lac[italic]Drd2[sup]-/-[/sup][/italic] mice had similar body length than [italic]Drd2[sup]loxP/loxP[/sup][/italic] mice, but increased gonadal and retroperitoneal fat pads, as well as heavier livers (142%, 122% and 34% increase, respectively). Daily food intake was also increased in lac[italic]Drd2[sup]-/- [/sup][/italic]compared to [italic]Drd2[sup]loxP/loxP [/sup][/italic]mice (3.47[plusmn]0.10 g vs. 3.02[plusmn]0.17 g). Hypothalamic expression of genes associated to food intake showed that precursor of orexin ([italic]Ppo)[/italic] mRNA decreased in [italic]Drd2[sup]-/- [/sup][/italic]mice but not in lac[italic]Drd2[sup]-/-[/sup][/italic]. In contrast, hypothalamic [italic]Npy[/italic] mRNA was increased only in lac[italic]Drd2[sup]-/-[/sup][/italic] female mice. Thus, the increase in food intake, body weight and adiposity observed in lac[italic]Drd2[sup]-/-[/sup][/italic] females in comparison to [italic]Drd2[sup]-/-[/sup][/italic] mice highlights the importance of functional central D2Rs in the orexigenic effect promoted by high prolactin levels and suggest that the increased levels of [italic]Npy[/italic] mRNA found in lac[italic]Drd2[sup]-/-[/sup][/italic] female mice are probably involved in these phenotypes.[br][br]Nothing to Disclose: GML, MIPM, DB-V, MR 2012-06-23T12:15:00 371 2012-06-23T00:00:00 1899-12-30T12:15:00 442 35 66 OR02-5 OR20-01 Saturday 11 2012


12 ENDO12L_OR02-6 ORAL SESSION: Development [amp] Modulation of Hypothalamic Neuropeptidergic Neurons (11:15 AM-12:45 PM) Lateral Hypothalamic Neurotensin Neurons Act by Multiple Neural Pathways and Control Water Intake and Activity Gina M Leinninger, Paulette Goforth, Adriana Gata Garcia, Lyndsay M Christensen, Leslie Satin, Martin G Myers, Jr University of Michigan, Ann Arbor, MI; University of Michigan, Ann Arbor, MI The lateral hypothalamic area (LHA) integrates diverse physiologic stimuli with appropriate behavioral outputs (including feeding, drinking and locomotor behavior) to maintain homeostasis, but specific roles for individual populations of LHA neurons remain unresolved. We investigated the function of GABAergic LHA neurons that contain the neuropeptide Neurotensin (Nts). LHA Nts neurons are activated by dehydration, while the adjacent population of Orexin/hypocretin (OX)-containing neurons are inhibited by water deprivation. We injected an AAV that mediates the cre-dependent expression of the D3q Designer Receptor Activated Exclusively by Designer Drugs (DREADDs) into the LHA of Nts-cre mice, permitting the selective activation of LHA Nts neurons in response to the DREADD ligand, clozapine-N-oxide (CNO). Consistent with our previous data demonstrating the importance of LHA Nts neurons for the regulation of OX neurons by leptin, CNO-mediated activation of DREADD-expressing LHA Nts neurons inhibited a population of OX neurons. This DREADD-mediated activation of LHA Nts neurons promoted voracious drinking and increased locomotor activity, although food intake was not changed. While LHA Nts neurons synapse on LHA OX neurons and ventral tegmental area (VTA) dopamine (DA) neurons, numerous VTA DA neurons express Neurotensin Receptor-1 (NtsR1), but OX neurons do not. As VTA Nts promotes locomotor activity, our data suggest that LHA Nts neurons inhibit LHA OX neurons by GABA release and activate VTA dopamine neurons via Nts to modulate ingestive behaviors and locomotion.[br][br]Sources of Research Support: NIH R01 DK78056 (MGM) and NIH K99 DK090101 (GML).[br][br]Nothing to Disclose: GML, PG, AGG, LMC, LS, MGM 2012-06-23T12:30:00 371 2012-06-23T00:00:00 1899-12-30T12:30:00 1539 35 67 OR02-6 OR20-01 Saturday 12 2012


13 ENDO12L_OR03-1 ORAL SESSION: Cardiometabolic Risk (11:15 AM-12:45 PM) The Intake of Fiber Suppresses the High-Fat High-Carbohydrate Meal-Induced Endotoxemia, Oxidative Stress and Inflammation Husam Ghanim, Sandeep Dhindsa, Manav Batra, Kelly Green, Sanaa Abuaysheh, Ajay Chaudhuri, Paresh Dandona State University of New York at Buffalo, Buffalo, NY Our previous work has shown that the intake of a high fat high carbohydrate (HFHC) meal results in an increase in endotoxin (LPS) and LPS binding protein (LBP) concentrations concomitant with an increase in the expression of TLR-4, the receptor for LPS, and CD-14,which facilitates the binding of LPS to its receptor. In addition, there is an increase in ROS generation, NF[kappa]B binding and the expression of TNF[alpha] and IL-1[beta]. Our work has also demonstrated that an equicaloric meal rich in fiber and fruit (American Heart Association (AHA) recommended meal) does not induce these changes. We hypothesized, therefore, that the addition of fiber to a HFHC meal will prevent the changes induced by HFHC meal. Eight fasting normal subjects (BMI[lt]25Kg/m2) were given 900 Calorie meals of either HFHC or AHA or HFHC with additional fiber (30g) (Fiber One Original) on 3 sequential visits one week apart in a randomized crossover design. As expected, the HFHC meal induced an increase in LPS concentrations (by 69[plusmn]14%) with an increase in ROS generation (by 120[plusmn]24%) and the expression of TLR-4, CD14 and IL-1[beta] (by 71[plusmn]14%, 86[plusmn]14% and 127[plusmn]14%, respectively; p[lt]0.05 for all) in MNC. The AHA meal did not induce any of these changes. The addition of fiber to HFHC meal significantly reduced the increase in LPS concentrations (by 58[plusmn]10%; p[lt]0.05), ROS generation (by 47[plusmn]14%; p[lt]0.05) and the expression of TLR-4, CD14 and IL-1[beta] (by 37[plusmn]11%, 46[plusmn]12% and 52[plusmn]15%, respectively; p[lt]0,05 for all) observed with the HFHC meal alone. Additionally, the intake of fiber with the HFHC meal reduced postprandial glucose excursion and increased insulin concentrations compared to HFHC meal alone. We conclude that the addition of fiber to a pro-inflammatory HFHC meal has a beneficial anti-inflammatory and metabolic effect. Thus, the fiber content of the AHA meal may account for its non-inflammatory nature. Since ROS, TLR-4 and IL-1[beta] potentially interfere with insulin signal transduction and are known to be pro-inflammatory in the arterial wall, this action of dietary fiber may contribute to its benefits in the prevention of insulin resistance, type 2 diabetes and atherogenesis.[br][br]Disclosures: PD: Speaker, Novo Nordisk. Nothing to Disclose: HG, SD, MB, KG, SA, AC 2012-06-23T11:15:00 360 2012-06-23T00:00:00 1899-12-30T11:15:00 1997 36 68 OR03-1 OR10-01 Saturday 13 2012


14 ENDO12L_OR03-2 ORAL SESSION: Cardiometabolic Risk (11:15 AM-12:45 PM) Plasma 25-Hydroxyvitamin D Concentration and Prevalence of Metabolic Syndrome in the Diabetes Prevention Program Randomized Controlled Trial Joanna Mitri, Jason Nelson, Cheryl Garganta, David Nathan, Frank Hu, Bess Dawson-Hughes, Anastassios G Pittas Tufts Medical Center, Boston, MA; Institute for Clinical Research and Health Policy Studies, Boston, MA; Floating Hospital for Children at Tufts Medical Center, Boston, MA; Massachusetts General Hospital and Harvard Medical School, Boston, MA; Harvard School of Public Health Boston, MA and Channing Laboratory, Brigham and Women[apos]s Hospital, Boston, MA; Tufts University, Boston, MA Background: Low vitamin D status, assessed by blood 25-hydroxyvitamin D (25OHD) concentration, has been associated with cardiometabolic disease, including metabolic syndrome and type 2 diabetes but results are not consistent. The association of 25OHD and metabolic syndrome has not been studied in a multi-ethnic population at risk for diabetes and there are limited reports on the association between 25OHD concentration and non-traditional components of metabolic syndrome.[br]Objective: To investigate the association of plasma 25OHD concentration with metabolic syndrome and its individual components, and markers of insulin secretion and sensitivity in the at-risk-for-diabetes population of the Diabetes Prevention Program (DPP).[br]Design, setting and patients: Cross-sectional analysis using baseline data from the placebo and lifestyle intervention arms of the DPP (N=2040). The metformin arm was excluded to minimize the cost associated with measurement of 25OHD.[br]Outcomes: Metabolic syndrome as defined according to the modified criteria from the National Cholesterol Education Program[apos]s Adult Treatment Panel; individual components of metabolic syndrome (fasting plasma glucose, triglycerides and HDL cholesterol levels, waist circumference and blood pressure); insulin sensitivity, insulin secretion and disposition index (based on oral glucose tolerance test).[br]Results: After multivariate adjustment, including age, gender, race, site location, month of blood draw, body mass index, smoking, alcohol consumption, C-reactive protein, ultraviolet index and physical activity, participants in the highest tertile of vitamin D (median 25OHD concentration 30.6 ng/mL) had a lower risk of prevalent metabolic syndrome (odds ratio 0.62; 95%CI 0.45; 0.84) compared to participants in the lowest tertile (median 25OHD concentration 12.1 ng/mL). Those in the highest tertile of vitamin D status had smaller waist circumference, higher HDL, and lower fasting plasma glucose. Higher plasma 25OHD concentration was associated with higher insulin sensitivity and lower insulin secretion while there was no association with disposition index.[br]Conclusion: In a multi-ethnic population at high risk for diabetes, higher plasma 25OHD concentration was inversely associated with metabolic syndrome.[br][br]Sources of Research Support: Diabetes Program Precention Research Group. By research grants R01DK76092 and R01DK79003 (to AGP) from the National Institute of Diabetes and Digestive and Kidney Disease, the Office Of The Director - National Institutes of Health, and the National Institutes of Health Office of Dietary Supplements; UL1RR025752 (to Tufts Medical Center, Boston, MA) from the National Center for Research Resources; The US Department of Agriculture Agreement 58-1950-9001 (to BDH); The Marilyn Fishman Grant for Diabetes Research grant (to JM) from the Endocrine Fellows Foundation; research grant UO1DK48489 from the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) of the National Institutes of Health to the DPP clinical centers and the Coordinating Center for the design and conduct of the DPP study; The Southwestern American Indian Centers were supported directly by the NIDDK, including its Intramural Research Program, and the Indian Health Service; The General Clinical Research Center Program, National Center for Research Resources, supported data collection at many of the clinical centers; Funding was also provided by the National Institute of Child Health and Human Development, the National Institute on Aging, the National Eye Institute, the National Heart Lung and Blood Institute, the Office of Research on of Women[apos]s Health, the National Center for Minority Health and Human Disease, the Centers for Disease Control and Prevention, the Indian Health Service, and the American Diabetes Association; Lipha (Merck-Sante) provided medication. LifeScan Inc., Merck-Medco Managed Care, Inc., and Merck and Co. donated materials, equipment, or medicines for concomitant conditions.[br][br]Nothing to Disclose: JM, JN, CG, DN, FH, BD-H, AGP 2012-06-23T11:30:00 360 2012-06-23T00:00:00 1899-12-30T11:30:00 993 36 69 OR03-2 OR10-01 Saturday 14 2012


15 ENDO12L_OR03-3 ORAL SESSION: Cardiometabolic Risk (11:15 AM-12:45 PM) The Effects of Caloric Restriction on Fetuin-A and Cardiovascular Risk Factors in Rats and Humans: A Randomized Controlled Trial Ho Cheol Hong, Jae Hee Ahn, Nam Hoon Kim, Yoonjung Kim, Hae Yoon Choi, Joo Hyung Kim, Chai Ryoung Eun, Sae Jeong Yang, Hye Jin Yoo, Hee Young Kim, Ji A Seo, Nan Hee Kim, Sei Hyun Baik, Dong Seop Choi, Kyung Mook Choi Korea University, Seoul, Korea The secreted liver protein fetuin-A is associated with insulin resistance, metabolic syndrome, type 2 diabetes, and atherosclerosis. We examined the effect of caloric restriction (CR) on fetuin-A level and concomitant changes in hepatic steatosis and cardiovascular risk factors in rats and humans.[br]We performed a randomized controlled clinical trial to examine the circulating fetuin-A level and cardiovascular risk parameters including visceral fat area (VFA), atherogenic lipid profile, inflammatory markers, adipokines and brachial artery endothelial function in 76 overweight women with type 2 diabetes before and after 12 weeks of CR. In addition, the effects of CR on hepatic steatosis and fetuin-A mRNA expression were evaluated in obese OLETF rats, an animal model of obesity and type 2 diabetes.[br]Circulating fetuin-A levels were significantly decreased after 12 weeks of CR, accompanied by improvements in VFA, blood pressure, glucose, lipid profiles, and liver function. The CR group also showed significantly deceased apolipoprotein B, leptin, and insulin resistance compared to those in the control group, although endothelial function were not different. Multiple regression analysis showed that the changes in fetuin-A were independently associated with CR and changes in hsCRP, adiponectin, and HOMA-IR ([italic]R[/italic][sup]2[/sup] = 24.6%). Moreover, CR significantly reduced hepatic steatosis and fetuin-A expression, as well as weight, glucose, total cholesterol and triglyceride levels in OLETF rats.[br]CR significantly reduced the hepatic expression of fetuin-A and its circulating levels and improved several cardiovascular risk factors in obese rats and humans with type 2 diabetes.[br][br]Nothing to Disclose: HCH, JHA, NHK, YK, HYC, JHK, CRE, SJY, HJY, HYK, JAS, NHK, SHB, DC, KMC 2012-06-23T11:45:00 360 2012-06-23T00:00:00 1899-12-30T11:45:00 60 36 70 OR03-3 OR10-01 Saturday 15 2012


16 ENDO12L_OR03-4 ORAL SESSION: Cardiometabolic Risk (11:15 AM-12:45 PM) Low Serum Dehydroepiandrosterone Sulfate (DHEA-S) Is Independently Associated with Increased Risk of Cardiovascular Events in Elderly Men Asa Tivesten, Maria Nilsson, Daniel Carlzon, Liesbeth Vandenput, Magnus K Karlsson, Osten Ljunggren, Elizabeth Barrett-Connor, Dan Mellstrom, Claes Ohlsson University of Gothenburg, Gothenburg, Sweden; Lund University, Malm[ouml], Sweden; University of Uppsala, Uppsala, Sweden; University of California San Diego, La Jolla, CA [bold]Background:[/bold] Dehydroepiandrosterone (DHEA) is a sex hormone precursor from the adrenal cortex that circulates mainly as a sulfate ester (DHEA-S), which is desulfated within peripheral target tissues and subsequently exerts biological effects via conversion into active sex steroids (e.g., testosterone and estradiol). Several studies have addressed the association between DHEA-S levels and cardiovascular (CV) disease, but most are small cross-sectional or mortality studies and findings are inconsistent. The aim of the present study was to test the hypothesis that serum DHEA-S is an independent predictor of major fatal and nonfatal CV events in a large population-based cohort of elderly men followed for 5 years.[br][bold]Methods: [/bold]We used liquid/gas chromatography-mass spectrometry to analyze baseline levels of DHEA and DHEA-S in the prospective population-based MrOS Sweden study (2416 men, age 69-81 years). Complete CV clinical outcomes were available from central Swedish registers.[br][bold]Results: [/bold]During follow-up, 485 fatal and nonfatal CV events occurred. Both DHEA and DHEA-S levels were inversely associated with the risk of CV events. Men in the lowest quartile of DHEA-S had increased risk of major CV events compared with men in pooled quartiles 2-4 (age-adjusted hazard ratio [HR] 1.25, 95% confidence interval [CI] 1.03-1.53). This association remained significant after adjustment for traditional CV risk factors and serum testosterone and estradiol levels, and was not materially changed in analyses excluding 634 men with known CV disease at baseline or men whose CV events occurred during the first years of follow-up (follow-up 2.6 years or less; n=241). Men in the lowest quartile of both DHEA and DHEA-S, compared with men in quartile 2-4 of both, showed increased risk of any major CV event (HR 1.34, 95% CI 1.06-1.70), of coronary heart disease (HR 1.41, 95% CI 1.05-1.89) and cerebrovascular (HR 1.41, 95% CI 1.00-1.99) events.[br][bold]Conclusion:[/bold] Low serum DHEA-S independently predicted increased 5-year risk of CV events in older men.[br][br]Sources of Research Support: This study was supported by the Swedish Research Council, the Swedish Foundation for Strategic Research, The Avtal om L[auml]karutbildning och Forskning research grant in Gothenburg, the Swedish Heart-Lung Foundation, the Marianne and Marcus Wallenberg Foundation, the Lundberg Foundation, the Torsten and Ragnar S[ouml]derberg[acute]s Foundation, the [Aring]ke Wiberg Foundation and the Novo Nordisk Foundation.[br][br]Nothing to Disclose: AT, MN, DC, LV, MKK, OL, EB-C, DM, CO 2012-06-23T12:00:00 360 2012-06-23T00:00:00 1899-12-30T12:00:00 472 36 71 OR03-4 OR10-01 Saturday 16 2012


17 ENDO12L_OR03-5 ORAL SESSION: Cardiometabolic Risk (11:15 AM-12:45 PM) A Novel PPAR[gamma] Ligand, GQ16,decreases Inflammation and Atherosclerosis in a Mouse Model of the Metabolic Syndrome Alan Richard Collins, Laurie J Minze, Tuo Deng, Lin Jianxin, Maricela Ramirez, Rajagopalan Senapathy, Kevin Phillips, Joey Z Liu, Christopher J Lyon, Anders L Berkenstam, Paul Webb, Willa Ann Hsueh The Methodist Hospital Research Institute, Houston, TX; The Methodist Hospital Research Institute, Houston, TX Current ligands to peroxisomal proliferator activated receptor gamma (PPAR[gamma]) used to treat diabetes are highly useful, but have limiting detrimental side effects. New compounds are being sought that retain the beneficial anti-inflammatory and antioxidant effects without side effects. New ligands which specifically inhibit cdk5 phosporylation of PPAR[gamma] (SPPARMs) are being developed, but they have little anti-inflammatory effect compared to thiazolidinediones (TZDs). We previously characterized a mouse model of the metabolic syndrome, the male middle aged (12 months old) low density lipoprotein receptor deficient (Ldlr[sup]-/-[/sup]) mouse fed 3 months Western diet, in which the TZD rosiglitazone (RSG) decreases accelerated atherosclerosis. Using this model, we tested a novel compound, GQ16, which binds PPAR[gamma] in an altered conformation compared to TZDs, but still inhibits cdk5 phosporylation. We placed mice on: 1) chow control, 2) Western diet (WD), 3) WD with 1.2 g rosiglitazone/kg food or 4) WD with 0.2 g GQ16/kg food. At sacrifice, atherosclerosis was determined by the en face method. All metabolic parameters were significantly increased in the mice fed WD compared to chow. There were no differences in the body weights, %body fat, or fasted glucose in any of the WD treatments. Treatment with either RSG (1.4 ng/ml, p[lt]0.0001) or GQ16 (3.1 ng/ml, p[lt]0.01) decreased plasma insulin levels compared to WD (6.1 ng/ml). Neither RSG nor GQ16 altered the plasma lipids significantly. Atherosclerosis was increased by WD (16.4% vs 5.8% respectively, p[lt]0.01), which was substantially attenuated by both RSG (6.3%, p[lt]0.01 vs WD alone) and GQ16 (9.1%, p[lt]0.01 vs WD alone). Lesion histology indicated that vessels of mice treated with GQ16 had an abundance of chondrocyte-like cells, indicating a more stable lesion, suggesting wound healing compared to vessels from RSG and untreated mice. Adipocytes isolated from epidydimal fat pads were assayed for inflammatory markers. Adiponectin expression decreased with WD, while interleukin 1[alpha] and 1[beta] as well as the MHC class II transactivator (CIITA) increased. Both RSG and GQ-16 preserved adiponectin expression and attenuated pro-inflammatory genes, IL-1[alpha] and IL-1[beta], and others, as well as CIITA. Ligands such as GQ16 with have strong potential in the development of novel PPAR[gamma] therapeutics to treat inflammatory and metabolic disorders.[br][br]Nothing to Disclose: ARC, LJM, TD, LJ, MR, RS, KP, JZL, CJL, ALB, PW, WAH 2012-06-23T12:15:00 360 2012-06-23T00:00:00 1899-12-30T12:15:00 2208 36 72 OR03-5 OR10-01 Saturday 17 2012


18 ENDO12L_OR03-6 ORAL SESSION: Cardiometabolic Risk (11:15 AM-12:45 PM) Diabetes Aggravates Reperfusion Injury by Reducing [underline]A[/underline]poptosis [underline]R[/underline]epressor with [underline]C[/underline]aspase Recruitment Domain (ARC) Expression Anastasia Susie Mihailidou, Thi Yen Loan Le, Mahidi Mardini, Anthony W Ashton Royal North Shore Hospital, Sydney, Australia; University of Sydney, Sydney, Australia; Royal North Shore Hospital [amp] University of Sydney, Sydney, Australia; Westmead Hospital, Sydney, Australia; Royal North Shore Hospital, Sydney, Australia Diabetes is the fastest growing disease worldwide and myocardial infarction (MI) is a major cause of death. Acute hyperglycaemia at the time of MI also predicts adverse cardiovascular outcomes. Using an [italic]ex-vivo[/italic] experimental myocardial infarction model we have previously reported aggravated infarct size and apoptosis in both a Type 2 animal model of diabetes (Zucker rat) and during acute hyperglycemia, although the mechanism was not defined. Since myocardial apoptosis correlates with unfavourable post-infarction left ventricular remodelling with progression to heart failure, we now target whether there is a common mechanism between acute and prolonged hyperglycemia. ARC ([underline]A[/underline]poptosis [underline]R[/underline]epressor with [underline]C[/underline]aspase recruitment domain) is an endogenous apoptosis repressor protein, highly expressed in cardiac tissue and is unique to other proteins, by inhibiting both death receptor (extrinsic) and mitochondrial (intrinsic) death pathways. [bold][italic]Aim: [/italic][/bold]To determine the expression and activity of antiapoptotic repressor protein, ARC, during acute and prolonged hyperglycemia. [bold][italic]Methods: [/italic][/bold]Male Sprague Dawley rats (SD) and obese Zucker rats (ZDF) were anesthetised, blood glucose measured and hearts isolated and subjected to regional ischemia (30min) followed by reperfusion (2.5hr). For acute exposure studies, 22 mM glucose was perfused 15 mins. prior to inducing ischemia and throughout reperfusion. ARC expression was detected by immunostaining myocardial sections and quantified. Infarct area was measured relative to the area-at-risk and total area of left ventricle. [bold][italic]Results:[/italic][/bold] Glucose levels for ZDF rats (19 [plusmn] 1 mM, n=10) were comparable to acute hyperglycemia levels. Ischemia-reperfusion (I-R) markedly reduced ARC levels [0.99 x 10[sup]6[/sup] [plusmn] 0.1, N=7 vs 2.15 x 10[sup]6[/sup] [plusmn] 0.3, N=7 (sham), p[lt]0.05]. Reduced ARC expression persisted during acute hyperglycemia (Gluc 22 mM) [0.63 x 10[sup]6[/sup] [plusmn] 0.09, N=6 vs 2.15 x 10[sup]6[/sup] [plusmn] 0.3, N=7 (sham), p[lt]0.05], while prolonged hyperglycemia levels in the ZDF rats, aggravated reduction of ARC levels due to I-R [0.44 x 10[sup]6[/sup] [plusmn] 0.04, N=7 vs 0.99 x 10[sup]6[/sup] [plusmn] 0.1, N=7 (I-R), p[lt]0.05]. Preliminary studies to measure ARC activity show I-R induced loss of ARC expression. Both acute [58 [plusmn] 1.7%, N=8] and prolonged [49 [plusmn] 1.4%, N=7] hyperglycemia aggravated infarct area [39 [plusmn] 2%, N=8, (I-R alone), p[lt]0.05]. [bold][italic]Conclusion:[/italic][/bold] Our study identifies a novel mechanism in diabetes, whereby hyperglycemia aggravates reperfusion injury by degrading anti-apoptotic protein ARC, a master regulator of apoptosis.[br][br]Nothing to Disclose: ASM, TYLL, MM, AWA 2012-06-23T12:30:00 360 2012-06-23T00:00:00 1899-12-30T12:30:00 1180 36 73 OR03-6 OR10-01 Saturday 18 2012


37 ENDO12L_OR07-1 ORAL SESSION: New Insights into Clinical [amp] Molecular Diagnosis of Adrenal Tumors (11:15 AM-12:45 PM) Primary Pigmented Nodular Adrenocortical Disease (PPNAD) and Isolated Micronodular Adrenocortical Disease (iMAD) Are Not Only Genetically but Also Biochemically and Clinically Distinct Causes of Pediatric ACTH-Independent Cushing Syndrome Evgenia Gourgari, Anya Rothebuhler, Margaret Keil, Anelia Horvath, Constantine Stratakis National Institute of Child Health and Human Development, Bethesda, MD; National Institute of Child Health and Human Development, Bethesda, MD Endogenous Cushing syndrome (CS) is caused by excess adrenal glucocorticoid secretion that is adrenocorticotropin(ACTH)-dependent or -independent. In both adults and children, adrenocortical lesions causing CS include the cortisol-producing adenoma, the rare adrenocortical cancer, and bilateral adrenocortical hyperplasias (BAHs). BAHs can be further divided in micronodular or macronodular hyperplasiaswith adrenal nodules smaller or larger than 1 cm, respectively. Micronodular BAHs can be further divided in primary pigmented nodular adrenocortical disease (PPNAD) or various forms of isolated micronodular adrenocortical disease (iMAD). PPNAD is characterized by internodular atrophy and intense nodular pigment, mostly due to the deposition of lipofuscin, while iMAD has a variable picture of hyperplasia of zona fasciculata and is characterized by the relative absence of pigmentation. Patients with PPNAD show paradoxical stimulation of urinary 17-hydroxy steroids (17-OHS) and free cortisol (UFC) versus those with macronodular hyperplasias and/or cortisol-producing adenomas during the course of Liddle[apos]s test. It is unknown whether iMAD consistently shares this feature with PPNAD. Genetically, the two disorders are different, since most patients with PPNAD have germline mutations of PRKAR1A, whereas iMAD is due to mutations in mostly unknown genes, with the minority of the cases caused by germline defects of the phosphodiesterase (PDE) genes PDE11A and PDE8B. We studied patients that were seen at NIH during the last 30 years, all below the age of 21 years. We identified 10 patients with iMAD and 21 patients with PPNAD that had undergone Liddle[apos]s test before bilateral adrenalectomy for CS. We found statistically significant differences of iMAD versus PPNAD when comparing age at surgery (mean age 8.54 vs 14.07 years), BMI z score (2.43 vs 1.47, p=0.001), baseline 24hUFC/m2 (mean 351 vs 98mcg/m2/day, p=0.02). No difference was observed in sex distribution, baseline 24h urine17OH/gr cr and response to Liddles test. We conclude that both PPNAD and iMAD can be diagnosed by the [ldquo]paradoxical[rdquo] response to Liddle[apos]s test; however, they are genetically, histopathologically, and even clinically different disorders.[br][br]Nothing to Disclose: EG, AR, MK, AH, CS 2012-06-23T11:15:00 372 2012-06-23T00:00:00 1899-12-30T11:15:00 2085 40 92 OR07-1 OR01-01 Saturday 37 2012


38 ENDO12L_OR07-2 ORAL SESSION: New Insights into Clinical [amp] Molecular Diagnosis of Adrenal Tumors (11:15 AM-12:45 PM) Impact of Mutations in the Selectivity Filter of the KCNJ5 Channel on the Subtyping of Primary Aldosteronism with Adrenal Vein Sampling Teresa M Seccia, Maniselvan Kuppusamy, Giulio Ceolotto, Brasilina Caroccia, Maria V Cicala, Franco Mantero, GianPaolo Rossi University of Padua, Padua, Italy Background and aim. In primary aldosteronism (PA) mutations affecting the selectivity filter of the KCNJ5 K+ channel (Choi, Science 2011) were suggested to be associated with a more prominent aldosterone excess as assessed by peripheral plasma aldosterone concentrations (PAC)(Boulkroun, Hypertension 2012). Hence, we hypothesize that these mutations, being somatic, could determine a higher aldosterone secretion from the adrenal gland with the aldosterone-producing adenoma (APA) and therefore could affect the adrenal vein sampling (AVS) results. We therefore compared the lateralization index (LI) and the cortisol-corrected PAC in the adrenal vein draining the APA and the contralateral side between APA with (KCNJ5mut) and without (KCNJ5wt) the mutation.[br]Patients and Methods: Consecutive APA patients (n=86) undergoing AVS underwent sequencing of APA and germinal DNA to seek for the presence of the KCNJ5 mutations (Boulkroun, Hypertension 2012). We calculated the ratio of cortisol(PCC)-corrected PAC from the APA side (PAC/PCC)APA and of the PAC/PCC in the inferior vena cava (PAC/PCC)IVC and the lateralization index (LI) was calculated as the ratio of (PAC/PCC)APA and (PAC/PCC) at the contralateral side.[br]Results. The overall prevalence rate of G151R, L168R and T158A mutations was 34%. The KCNJ5mut and KCNJ5wt groups did not differ for gender, age, serum K+ levels, and ARR and blood pressure fall post-adrenalectomy. At baseline the KCNJ5mut group showed significantly higher (p[lt]0.03) PAC and ARR, and lower PRA (p[lt]0.03) than KCNJ5wt group, which suggested a more prominent aldosterone excess. This was reflected in a higher (PAC/PCC)APA/(PAC/PCC)IVC ratio and a higher LI in the KCNJ5mut than in the KCNJ5wt group. Accordingly, the LI was higher in the former than in the latter group (29.3[plusmn] 6.7 vs 16.7[plusmn]3.9, p[lt] 0.02).[br]Conclusions. The APA carrying the KCNJ5 mutations in the selectivity filter are characterized by a secretion of aldosterone from the tumor higher than APA wild type. As this translated into a higher LI at AVS, the presence of KCNJ5 mutations is likely to increase the accuracy of the subtyping of PA by means of AVS.[br][br]Nothing to Disclose: TMS, MK, GC, BC, MVC, FM, GR 2012-06-23T11:30:00 372 2012-06-23T00:00:00 1899-12-30T11:30:00 1886 40 93 OR07-2 OR01-01 Saturday 38 2012


39 ENDO12L_OR07-3 ORAL SESSION: New Insights into Clinical [amp] Molecular Diagnosis of Adrenal Tumors (11:15 AM-12:45 PM) Mitotane Induces a Mitochondrial Cytochrome c Oxidase Defect: A Novel Pharmacological Action for Adrenocortical Carcinoma Management Segolene Hescot, Abdelhamid Slama, Angelo Paci, Herve Remy, Rita Chadarevian, Severine Trabado, Anne Lombes, Jacques Young, Eric Baudin, Marc Lombes Faculty Medicine Paris SUd, Le Kremlin Bic[ecirc]tre, France; CHU Bicetre, Le Kremlin Bic[ecirc]tre, France; Institut Gustave Roussy, Villejuif, France; HRA-PHARMA France, Paris, France; Institut Cochin, Paris, France; CHU Bicetre, Le Kremlin Bic[ecirc]tre, France; Institut Gustave Roussy, Villejuif, France Mitotane (o,p[apos]DDD) is the most effective therapeutic strategy for adrenocortical carcinoma (ACC). Yet the molecular mechanisms of mitotane action remain unclear despite its well-established antisecretory and antiproliferative properties. Mitochondrial (mt) impact of mitotane has been previously suggested but not yet fully validated. We examined functional consequences of mitotane exposure on mitochondrial steroidogenesis, respiratory chain activity and biogenesis, using human adrenocortical secreting H295R and non-secreting SW13 cells. We first confirmed that mitotane exposure inhibits cell proliferation in a dose- and time-dependent manner. We were however unable to detect mitotane metabolites, o,p[apos]DDA and o[apos]p[apos]DDE, in cultured cells supernatant but could not rule out an intracellular metabolic transformation. We demonstrated that mitotane drastically reduces cortisol and 17-hydroxyprogesterone secretions by 70%, at a 50 [mu]M (14 mg/L) concentration, considered as the therapeutic plasma level threshold. This was accompanied by significantly decreased expression of genes encoding mitochondrial proteins involved in steroidogenesis (STAR, CYP11B1, CYP11B2). In both H295R and SW13 cells, 50 [mu]M mitotane induced a significant and reproducible 50% decrease of the maximum velocity of the respiratory chain complex IV (cytochrome c oxidase, COX) activity. This was associated with significantly decreased expression of both the mtDNA-encoded COX2 and the nuclear DNA encoded COX4 COX subunits, and with an important reduction of the whole COX steady state expression, as revealed by Blue Native PAGE. Enhanced mitochondrial biogenesis was shown by an increased mt DNA content and enhanced expression of the transcriptional regulator PGC1[alpha], consistent with an adaptive and compensatory mechanism against mitochondrial defect. Mitotane exposure also triggered a morphologic shift from filamentous to punctuate aspects of mitochondrial network, providing additional support for alteration in the mitochondrial compartment dynamics.[br]Collectively, our results provided direct evidence that mitotane induces mitochondrial respiratory chain defect. Better understanding of mitotane pharmacology should enable us to identify predictive factors of efficacy and toxicity.[br][br]Sources of Research Support: Grants from Inserm, Univ Paris-Sud, HRA-Pharma.[br][br]Nothing to Disclose: SH, AS, AP, HR, RC, ST, AL, JY, EB, ML 2012-06-23T11:45:00 372 2012-06-23T00:00:00 1899-12-30T11:45:00 913 40 94 OR07-3 OR01-01 Saturday 39 2012


40 ENDO12L_OR07-4 ORAL SESSION: New Insights into Clinical [amp] Molecular Diagnosis of Adrenal Tumors (11:15 AM-12:45 PM) Comparing Biochemical and Radiological Characteristics in Pheochromocytoma in Patients with Less Than Four Fold Elevated Metanephrines to Those with Higher Levels Subramanian Kannan, Andrei Purysko, Erick Remer, Allen Sipperstein, Eren Berber, Amr Fergany, Charles Faiman, Amir Hamrahian, Emmanuel Bravo Cleveland Clinic Foundation, Cleveland, OH; Cleveland Clinic Foundation, Cleveland, OH; Cleveland Clinic Foundation, Cleveland, OH; Cleveland Clinic Foundation, Cleveland, OH [bold]Background:[/bold] Pheochromocytoma (Pheo) needs to be ruled out in patients with suggestive symptomatology, adrenal incidentaloma and predisposing hereditary conditions. Tumor size shows a strong positive correlation with summed plasma (PL) or urinary (Ur) Metanephrines (METs). Correlation between CT attenuation (HU) and PL and/or Ur METs has not been described. A 4 fold elevation or higher of upper limits of normal (ULN) of PL or Ur METs is usually diagnostic for Pheo. A combination of PL or Ur METs and radiological features may be helpful in a timely diagnosis of Pheo in patients with METs [lt]4 fold ULN.[br][bold]Aim:[/bold] To compare Clinical and Radiological features in patients with Pheo who had PL and/or Ur METs [ge] 4 fold ULN (G1) and those with levels [lt] 4 fold ULN (G2).[br][bold]Methodology:[/bold] Among 252 patients with pathologically proven pheochromocytoma, we identified 105 patients who had images and at least one set of PL or Ur METs available for review (1997-2011). Metastatic Pheos were excluded. MET and NorMET were expressed as ratios above the ULN.[br][bold]Results: [/bold]The median (range) age and F/M sex ratios for G1 (n=83) and G2 (n=22) were 51 (19-84) years, 44/39; and 50 (24-73) years, 11/11 respectively. The percentage of patients with predisposing family history (G2 32% vs G1 12%, p 0.027) was different but similar for incidentally detected Pheo (G2 73% vs G1 48%, p 0.22). Extra-adrenal Pheo was exclusively seen in G1 (11%). There was a stronger correlation between radiological size of the tumor and summed ratios of PL METs (p [lt]0.001) and Ur METs (p[lt] 0.001) in G1 compared to G2 (p 0.14 and p 0.019). There was no correlation between PL or Ur METs and pre-contrast HU in each group. The radiological tumor size (cm) and the non-contrast CT HU for Gr 1 and Gr 2 was 4.9 (2-22) cm, 35.5 (17-59) HU; and 3 (0.9-5) cm;, 39 (25-52) HU, respectively (p [lt]0.001; p 0.65). The distribution of the radiological features in both groups are as follows (%): Heterogeneity (83 vs. 43 p 0.005), Necrosis (73 vs. 50 p 0.11), Hypervascularity (61 vs 56, p 0.99), Calcification (7 vs. 0, p 0.58), Hemorrhage (11 vs 0, p 0.58) and Cystic changes (69 vs. 50, p 0.22).[br][bold]Discussion:[/bold] We did not identify any tumor [gt] 5cm in size with PL or Ur METs [lt]4 fold ULN. None of the extr-adrenal tumors had elevations [lt] 4 fold ULN. Pheos in both groups had a pre-contrast HU [ge]17.[br][bold]Conclusion:[/bold] Patients with PL or Ur METs [lt] 4 fold ULN were more likely to have smaller tumors, predisposing family history and homogenous tumors on imaging.[br][br]Nothing to Disclose: SK, AP, ER, AS, EB, AF, CF, AH, EB 2012-06-23T12:00:00 372 2012-06-23T00:00:00 1899-12-30T12:00:00 1656 40 95 OR07-4 OR01-01 Saturday 40 2012


41 ENDO12L_OR07-5 ORAL SESSION: New Insights into Clinical [amp] Molecular Diagnosis of Adrenal Tumors (11:15 AM-12:45 PM) Genotype-Specific Correlations between Respiratory Chain Enzyme Activities, Succinate Accumulation and Catecholamine Content in Pheochromocytoma and Paraganglioma: An [italic]In Vitro[/italic] 1H-NMR Spectroscopy Study Jyotsna Upendra Rao, Udo F Engelke, Richard J Rodenburg, Ron A Wevers, Karel Pacak, Benno Kusters, Angelina G Goudswaard, Jacques W Lenders, Ad R Hermus, Arjen R Mensenkamp, Graeme Eisenhofer, Dirk H Kunst, Fred C Sweep, Henri J Timmers Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands; National Institute of Child Health and Human Development, Bethesda, MD; University Hospital Carl Gustav Carus, Dresden, Germany Background: Direct implication of mitochondrial enzymes in the tumorigenesis of pheochromocytoma and paraganglioma (PGL) came with the discovery of heterozygous germline mutations in succinate dehydrogenase (SDH) subunits causing familial paraganglioma syndromes. SDH catalyses the oxidation of succinate to fumarate in Krebs cycle and forms complex II of the mitochondrial respiratory chain. Reduced expression of components of respiratory chain complexes (RCC) and Krebs cycle has been observed in type I sporadics and patients with VHL (von Hippel Lindau) and SDHx mutations. Altered expression of components of RCCs, Krebs cycle and glycolysis in PGLs caused by SDHx and VHL mutations appears to be related to hypoxia/angiogenesis pathways of tumorigenesis through the stabilization of HIFs. Further, differences in energy metabolism in PGL may also translate into distinct profiles of catecholamine production, metabolism and secretion.[br]Objective: The aim of the study was to examine the relationship between RCC activities and succinate and catecholamine content of PPGLs across different genotypes.[br]Methods: 72 primary PGL tissues collected from patients with the following genotypes were investigated: SDHB (n=11), SDHD (n=7), SDHAF2 (n=2), VHL (n=7), NF1 (n=7), MAX (n=2), RET (n=14) and sporadic (n=22). Patients whose genotype was tested negative for SDHB, SDHC, SDHD, RET and VHL were considered as sporadics. The tumor tissue homogenates prepared in SEF buffer (10% w/v) were assayed for RCCs (complex I-IV) on Konelab20XT autoanalyzer using protocols reported before (1). Succinate and catecholamine levels in these tissues were measured using 1H-NMR spectroscopy. For this purpose, the tissues were homogenized in MilliQ water (10% w/v) and deproteinized by ultrafiltration using a 10 kDa filter. The ultrafiltrates were analyzed further using 500MHz 1H-NMR spectrometer.[br]Results: Complex II activity in general was barely above the detection limit in tissues with SDHx mutations. Low complex II activity was significantly associated with vast accumulation of succinate and with norepinephrine dominant tumor catecholamine content. Variations in the activities of RCCs I, III and IV were also observed among the different genotypes.[br]Conclusion: Decreased complex II activity in SDHx-related PPGL correlates with succinate accumulation and a noradrenergic phenotype. Genotype specific changes in the energy metabolism appear to have an impact on catecholamine metabolism.[br][br](1) Rodenburg R., J Inherit Metab Dis 2011; 34:283.[br][br]Nothing to Disclose: JUR, UFE, RJR, RAW, KP, BK, AGG, JWL, ARH, ARM, GE, DHK, FCS, HJT 2012-06-23T12:15:00 372 2012-06-23T00:00:00 1899-12-30T12:15:00 224 40 96 OR07-5 OR01-01 Saturday 41 2012


42 ENDO12L_OR07-6 ORAL SESSION: New Insights into Clinical [amp] Molecular Diagnosis of Adrenal Tumors (11:15 AM-12:45 PM) Definition of a New Algorithm of Genetic Testing in Paraganglioma and Pheochromocytoma Syndromes Julia Katharina Bickmann, Stefanie Sollfrank, Heidi Rossmann, Arno Schad, Racha Hassoun, Carolin Neukirch, Brigitte Schneider-Ratzke, Konstantinos Papaspyrou, Wolf J Mann, Karl J Lackner, Matthias M Weber, Christian Fottner University Medical Center Mainz, Mainz, Germany; University Medical Center Mainz, Mainz, Germany; University Medical Center Mainz, Mainz, Germany; University Medical Center Mainz, Mainz, Germany; University Medical Center Mainz, Mainz, Germany [bold]Objective[/bold]: We intended to optimise routine genetic testing of 116 pheochromocytoma and paraganglioma index patients by extending the usual panel of genes (SDHB, SDHC, SDHD, VHL, RET-protooncogene) by other genes that have been reported to be involved in the pathogenesis of endocrine tumor syndromes and by complementing genetic testing by prospective clinical follow-ups of all of these patients.[br][bold]Design and Method[/bold]: We designed sequencing assays of the genes MAX, SDHAF2/SDH5, TMEM127, PHD2 and SDHA. Large rearrangements of the SDH genes were detected by a commercial MLPA (Multiplex Ligation Dependant Probe Assay). Pyrosequencing was applied in the screening of the only known SDHAF2/SDH5 mutation and in evaluating various potential SNPs in a healthy control population.[br][bold]Results and Conclusions[/bold]: Out of 116 index patients 62% suffered from paragangliomas, 36% from pheochromocytomas and 2% from both tumor types. In 34% (39/116) of cases the underlying mutations were identified (familial tumor syndromes). Mutations were found in the following genes (given as percentage of all tested patients): 5% RET, 3% VHL, 1% TMEM127, 8% SDHB, 5% SDHC, 11% SDHD, and 1% SDHA. There were 6 novel SDH mutations including a large deletion in SDHB and an amino acid exchange in codon 1 of SDHA. We are currently complementing the genetic analysis by SDHB and SDHA immunohistochemistry of tumour samples. Familial cases of paragangliomas and pheochromocytomas are more common than assumed in the past. Genetic characterisation of paraganglioma patients including the detection of genomic rearrangements is essential for genetic counseling. According to our data all paraganglioma and pheochromocytoma patients who have undergone tumor surgery should also have genetic testing irrespective of their family histories. Even oligosymptomatic patients (e.g. isolated glomus tumors) may have more seriously affected relatives who would benefit from an early diagnosis.[br][br]Nothing to Disclose: JKB, SS, HR, AS, RH, CN, BS-R, KP, WJM, KJL, MMW, CF 2012-06-23T12:30:00 372 2012-06-23T00:00:00 1899-12-30T12:30:00 1037 40 97 OR07-6 OR01-01 Saturday 42 2012


43 ENDO12L_OR08-1 ORAL SESSION: Osteoporosis Physiology [amp] Clinical Trials (11:15 AM-12:45 PM) Bone Anabolic Efficacy and Safety of BA058, a Novel Analog of hPTHrP: Results from a Phase 2 Clinical Trial in Postmenopausal Women with Osteoporosis Gary Hattersley, John Bilezikian, Jonathan Guerriero, Prasanna Kumar, Jose Zanchetta, C Richard Lyttle, Louis St L O[apos]Dea Radius Health, Cambridge, MA; Columbia University College of Physicians and Surgeons, New York, NY; M S Ramaiah Memorial Hospital, Bangalore, India; Instituto de Investigaciones Metabolicas, Buenos Aires, Argentina BA058 is a synthetic analog of hPTHrP (1-34) undergoing clinical development for the treatment of postmenopausal osteoporosis. This was a randomized, double blind, placebo-controlled Phase 2 study to determine BA058 safety and effects on BMD and bone markers. Healthy postmenopausal women with severe osteoporosis (ages 55 to 85 years) were randomized to placebo, BA058 20, 40 or 80 ug, or teriparatide (TP) 20 ug for 24 weeks in an international, multicenter study. Available patients who completed 24 weeks could participate in an additional 24 weeks of treatment. 184 (83%) of 221 patients completed 6 months of treatment. The mean percent change in lumbar spine BMD at 24 weeks was 1.6% with placebo, 2.9%, 5.2%, and 6.7% with BA058 20, 40 and 80 ug respectively, and 5.5% with TP. The difference from placebo was significant (p[lt]0.001) for BA058 40 and 80 ug and for TP. Further increases in lumbar spine BMD were seen during the extension phase (n=55), with a mean percent change at 48 weeks of 0.7% with placebo, 5.1%, 9.8%, 12.9% with BA058 20, 40 and 80 ug respectively, and 8.6% with TP. A marked, dose dependent increase in total hip BMD was also seen at 24 weeks, where the mean change was 0.4% with placebo, 1.4%, 2.0%, and 2.6% with BA058 20, 40, and 80 ug respectively, and 0.5% with TP. At 24 weeks the change in serum and urine markers was significant (p[le]0.05) from baseline for BA058 40 and 80 ug and for TP for PINP, BSAP, osteocalcin and CTX, and with TP for NTX. BA058 was generally well tolerated with treatment-emergent AEs reported in 164 (74%) of 221 patients, with similar proportions across treatment groups. The most commonly reported events were influenza (10.4%) and headache (10.4%). Seven patients (3.1%) discontinued due to an adverse event, and serious adverse events were reported in 1.4% patients, none were treatment related. Mean serum calcium levels were consistently higher with TP compared to BA058, with mean values continuing to rise with TP over time. The safety profile of BA058 was consistent through an additional 6 months of treatment and no new safety signals were observed. In conclusion, treatment with BA058 80 ug resulted in significant BMD gains at all measured sites. BA058 was well tolerated, with safety events comparable with blinded placebo and consistent with medical events in this population. The safety and efficacy data support selection of BA058 80 ug for further clinical development for the treatment of postmenopausal osteoporosis.[br][br]Disclosures: GH: Employee, Radius Health. JG: Employee, Radius Health. CRL: Employee, Radius Health. LSLO: Employee, Radius Health. Nothing to Disclose: JB, PK, JZ 2012-06-23T11:15:00 Theater A 2012-06-23T00:00:00 1899-12-30T11:15:00 443 41 98 OR08-1 OR05-01 Saturday 43 2012


44 ENDO12L_OR08-2 ORAL SESSION: Osteoporosis Physiology [amp] Clinical Trials (11:15 AM-12:45 PM) BMD Response in the Odanacatib Phase 2 Trial: Prespecified Threshold and Subgroup Analyses Michael McClung, Neil Binkley, Henry Bone, Paul Miller, Nadia Verbruggen, Carolyn DaSilva, Elizabeth Rosenberg, Albert Leung Oregon Osteoporosis Center, Portland, OR; University of Wisconsin, Madison, WI; Michigan Bone and Mineral Clinic, Detroit, MI; Colorado Center for Bone Research, Lakewood, CO; MSD Europe, Brussels, Belgium; Merck Sharp [amp] Dohme Corp, Whitehouse Station, NJ [bold]Background:[/bold] Odanacatib (ODN) is a cathepsin K inhibitor in phase 3 development for osteoporosis. Treatment with ODN 50 mg once weekly for 2 years resulted in placebo (PBO)-subtracted mean increases in lumbar spine (LS) and total hip (TH) bone mineral density (BMD) of 5.7% and 4.1% respectively, P[le]0.001 vs. placebo. Five years of continuous treatment with ODN 50 mg resulted in BMD increases from baseline of 11.9% at LS and 8.5% at TH, and sustained reductions in bone resorption markers. In contrast, levels of bone formation markers were near baseline.[br][bold]Objectives:[/bold]To evaluate BMD response by thresholds and in pre-specified subgroups.[br][bold]Methods:[/bold] 399 postmenopausal women with low baseline BMD were randomized to ODN 3, 10, 25, 50 mg or PBO weekly, plus vitamin D and calcium. Percent change from baseline in LS BMD was the primary endpoint. Prespecified BMD-change threshold analyses and subgroup analyses by age, ethnicity, and baseline BMD and biomarkers are reported for women who received weekly placebo or ODN 50 mg (the dose being investigated in the phase 3 trial).[br][bold]Results:[/bold] At the LS, 0%, 95%, 76%, and 41% of participants treated with ODN 50 mg weekly for 2 years had BMD changes of [le]-3%, [gt]0%, [gt]3%, and [gt]6%, respectively, compared with 24%, 48%, 18%, and 7% in the PBO group. At the TH, the proportions were 0%, 85%, 48%, and 19% in the ODN group and 33%, 41%, 8%, and 0% in the PBO group. Subgroup analysis of % changes from baseline in LS BMD after 2 years revealed no statistically significant treatment-subgroup interactions between women who at baseline were [lt]65 vs. [ge]65 years old, Caucasian vs. non-Caucasian, or above or below the median values of baseline LS, hip trochanter, or femoral neck BMD or baseline bone resorption markers CTX or DPD. However, there were greater treatment effects in LS BMD in women with lower baseline TH BMD, higher baseline bone resorption marker NTX, or higher baseline bone formation markers BSAP and P1NP (7.2%, 6.6%, 7.5%, 6.9%, respectively) compared with women with the opposite baseline characteristics (4.3%, 4.6%, 4.2%, 4.4%, respectively). TH BMD subgroup analyses showed similar trends of smaller magnitude.[br][bold]Conclusions: [/bold]Treatment with ODN 50 mg was considerably more likely than treatment with PBO to increase LS and TH BMD to each threshold level. Subgroup analysis of BMD change suggests that participants with lower BMD or higher bone turnover at baseline may have a greater % BMD increase with ODN treatment.[br][br]Disclosures: MM: Investigator, Amgen, Merck & Co., Takeda; Consultant, Amgen, Lilly USA, LLC, Merck & Co., Novartis Pharmaceuticals, Warner Chilcott Pharmaceuticals, Takeda; Speaker, Amgen, Lilly USA, LLC, Merck & Co., Novartis Pharmaceuticals, Warner Chilcott Pharmaceuticals.NB: Consultant, Merck & Co., Lilly USA, LLC, Tarsa; Investigator, Merck & Co., Amgen; Researcher, Tarsa. HB: Consultant, Amgen, Merck & Co., Takeda, Tarsa; Investigator, Amgen, Merck & Co., Nordic Bioscience A/S, Novartis Pharmaceuticals, Takeda, Tarsa; Speaker Bureau Member, Amgen. PM: Consultant, Merck & Co.; Investigator, Merck & Co. NV: Employee, Merck & Co. CD: Employee, Merck & Co. ER: Employee, Merck & Co. AL: Employee, Merck & Co. 2012-06-23T11:30:00 Theater A 2012-06-23T00:00:00 1899-12-30T11:30:00 1763 41 99 OR08-2 OR05-01 Saturday 44 2012


45 ENDO12L_OR08-3 ORAL SESSION: Osteoporosis Physiology [amp] Clinical Trials (11:15 AM-12:45 PM) Distinctive Role of 6-Month Teriparatide on Advanced Bisphosphonate-Related Osteonecrosis of the Jaw (BRONJ); Case-Control Study Kyoung Min Kim, Wonse Park, Hyung-Jun Kim, Woong Nam, Sung-Kil Lim, In Ho Cha, Yumie Rhee Yonsei University College of Medicine, Seoul, Korea; Yonsei University College of Dentistry, Seoul, Korea; Yonsei University College of Dentistry, Seoul, Korea Management of bisphosphonate-related osteonecrosis of the jaw(BRONJ) is a problematic in subset of patients. The objective was to determine whether combining teriparatide(TPTD) could overcome the difficulties in managing advanced cases of BRONJ patients. Twenty four subjects of advanced BRONJ, who did not recover from osteonecrosis with previous dental care including sequestrectomy, out of 119 BRONJ patients were chosen for the case-control study. Fifteen patients received daily 20 ug of TPTD injection subcutaneously along with conventional therapy for 6 months[TPTD group] and the other 9 patients were managed only with conservative care[non-TPTD group]. Bone turnover markers and hormones related to bone metabolism were analyzed at baseline, 1 and 6 months of treatment. Stages of BRONJ were assessed every month by two separate dentists using stages established by American Association of Oral and Maxillofacial Surgery. Outcome of BRONJ treatment was defined as [apos]not improved[apos] with no changes in stages of BRONJ after 6 months of treatment, [apos]moderately improved[apos] with one stage improvement and [apos]markedly improved[apos] with two stages upgrade or complete healing. The initial stages of BRONJ were mostly stage 2, 93.3%(14/15) in TPTD group and 88.9%(8/9) cases in non-TPTD groups, and the remainders of both groups were stage 3. The increment of C-telopeptide(CTx) was significantly higher in TPTD group compared to the level of non-TPTD group at 6 months (475.9[plusmn]334.9% vs. 0.2[plusmn]11.9%, [italic]p[/italic][lt]0.05). Osteocalcin also showed significant increase at 6 months in TPTD-group (254.1[plusmn]29.5% vs. -2.5[plusmn]7.5%, [italic]p[/italic][lt]0.05). After 6 months of treatment, 62.5% of TPTD group was classified as [apos]markedly improved[apos], 37.5% [apos]moderately improved[apos] without any case that did not improve, whereas final status of non-TPTD group ended up as 60%, [apos]moderately improved[apos] and 40%, [apos]not improved[apos] ([italic]p[/italic][lt]0.05 vs. TPTD group). Interestingly, 25(OH)D level showed lower tendency in [apos]moderately improved[apos] patients compared to [apos]markedly improved[apos] ones in TPTD group (15.3[plusmn]9.3 ng/dL vs. 29.7[plusmn]6.3 ng/dL, [italic]p[/italic]=0.056). Initial 1 month change of CTx had a strong positive correlation with 25(OH)D (r=0.649, [italic]p[/italic][lt]0.05) and a negative correlation with PTH (r=-0.670, [italic]p[/italic][lt]0.05). In conclusion, 6-month use of TPTD was superior on the resolution of advanced BRONJ compared to the conventional therapy via accelerated recovery of bone remodeling. Moreover, vitamin D insufficiency could be a risk factor for lesser response to the TPTD in BRONJ patients.[br][br]Nothing to Disclose: KMK, WP, H-JK, WN, S-KL, IHC, YR 2012-06-23T11:45:00 Theater A 2012-06-23T00:00:00 1899-12-30T11:45:00 748 41 100 OR08-3 OR05-01 Saturday 45 2012


46 ENDO12L_OR08-4 ORAL SESSION: Osteoporosis Physiology [amp] Clinical Trials (11:15 AM-12:45 PM) Sclerostin Levels and Bone Turnover Markers in Adolescents with Anorexia Nervosa and Healthy Adolescent Girls Alexander T Faje, Pouneh Fazeli, Debra K Katzman, Karen K Miller, Anne Breggia, Clifford J Rosen, Nara Mendes, Anne Klibanski, Madhusmita Misra Massachusetts General Hospital and Harvard Medical School, Boston, MA; Hospital for Sick Children and University of Toronto, Toronto, Canada; Maine Medical Center Research Institute, Portland, ME; Massachusetts General Hospital for Children and Harvard Medical School, Boston, MA Sclerostin is an important determinant of bone formation and resorption. Adult studies have shown that sclerostin is negatively regulated by estradiol. Adolescents with anorexia nervosa (AN) are hypogonadal and have low bone mineral density (BMD) and decreased levels of bone turnover markers. Sclerostin has not been examined in AN as a potential mediator of impaired bone metabolism.[br]Our study objectives were to (i) compare sclerostin in girls with AN and controls, (ii) assess associations with bone turnover markers, and (iii) examine effects of transdermal estradiol on sclerostin in AN.[br]A subset of adolescent girls with AN and normal-weight controls 13-18 years old were selected from a prior study based on their lumbar spine BMD Z-scores. The subset included all subjects with AN with a baseline lumbar BMD Z-score [lt] -0.5 and controls with lumbar BMD Z-scores between +1.0 and -1.0. 69 girls (44 with AN and 25 normal-weight controls) were studied at baseline. 22 AN girls with a bone age of [ge] 15 years were randomized to 100mcg transdermal estradiol (n=13) or placebo (n=9) in a double-blind study for 12 months. Subjects randomized to transdermal estradiol received medroxyprogesterone 2.5mg daily for 10 days each month. All subjects also received calcium carbonate (1200mg) and vitamin D (400 IU) daily.[br]Sclerostin levels did not differ in AN versus controls (0.422[plusmn]0.016 vs. 0.408[plusmn]0.018 ng/ml, p=0.58). Sclerostin correlated positively with P1NP and CTX in controls (r=0.49 and 0.55, p=0.01 and 0.004, respectively) but not in AN (r=0.09 and -0.17, p=0.62 and 0.34). Treatment with transdermal estradiol significantly improved lumbar BMD compared with placebo ([Delta] lumbar BMD: 0.035[plusmn]0.007 vs. 0.005[plusmn]0.012 g/cm[sup]2[/sup], p=0.01). However, changes in sclerostin over twelve months did not differ in girls randomized to estradiol or placebo (-0.101[plusmn]0.031 vs. -0.128[plusmn]0.042 ng/ml, p = 0.61).[br]Levels of sclerostin do not differ in girls with AN compared with normal-weight adolescents. The relationship between sclerostin and markers of bone turnover in normal-weight girls is disrupted in adolescent girls with AN. Despite an increase in BMD with transdermal estradiol administration in AN, estrogen does not impact sclerostin levels in this group. These findings imply (i) the presence of additional unspecified factors that regulate sclerostin in adolescents with AN, and (ii) that changes in sclerostin do not mediate the beneficial effects of transdermal estradiol on BMD in this group.[br][br]Sources of Research Support: R01 DK062249 and UL1 RR025758-01.[br][br]Nothing to Disclose: ATF, PF, DKK, KKM, AB, CJR, NM, AK, MM 2012-06-23T12:00:00 Theater A 2012-06-23T00:00:00 1899-12-30T12:00:00 88 41 101 OR08-4 OR05-01 Saturday 46 2012


47 ENDO12L_OR08-5 ORAL SESSION: Osteoporosis Physiology [amp] Clinical Trials (11:15 AM-12:45 PM) Denosumab for the Treatment of Men with Low Bone Mineral Density Ugis Gruntmanis, Eric Orwoll, Christence S Teglbjaerg, Bente L Langdahl, Roland Chapurlat, Edward Czerwinski, David L Kendler, Jean-Yves Reginster, Alan Kivitz, E Michael Lewiecki, Paul D Miller, Michael A Bolognese, Michael R McClung, Henry G Bone, Osten Ljunggren, Bo Abrahamsen, Yu-Ching Yang, Rachel W Wagman, Suresh Siddhanti, Andreas Grauer, Jesse W Hall, Steven Boonen Dallas Veterans Affairs Medical Center and University of Texas Southwestern Medical Center, Dallas, TX; Oregon Health and Science University, Portland, OR; Clinical and Basic Research, Ballerup, Denmark; Aarhus University Hospital, Aarhus, Denmark; Hopital Edouard Herriot, Lyon, France; Karkow Medical Center, Krakow, Poland; University of British Columbia, Vancouver, Canada; University of Liege, Liege, Belgium; Altoona Center for Clinical Research, Duncansville, PA; New Mexico Clinical Research and Osteoporosis Center, Alburquerque, NM; University of Colorado Health Science Center, Lakewood, CO; Bethesda Health Research Center, Bethesda, MD; Oregon Osteoporosis Center, Portland, OR; Michigan Bone and Mineral Clinic, Detroit, MI; Uppsala University, Uppsala, Sweden ; Copenhagen University Hospital Gentofte, Hellerup, Denmark; Amgen Inc, Thousand Oaks, CA; Amgen Inc, Thousand Oaks, CA; Leuven University, Leuven, Belgium [bold]Purpose: [/bold]The objective of this study was to evaluate the effect of denosumab (DMAb) on bone mineral density (BMD) in men with low BMD.[br][bold]Method: [/bold]This was a multicenter, randomized, double-blind, placebo-controlled study where subjects were randomized 1:1 to receive either 60 mg DMAb or placebo administered subcutaneously once every 6 months over a 12-month period. Subjects were male, and were included if they were [ge]30 and [le]85 yrs of age; had a BMD T-score [le]-2.0 and [ge]-3.5 at the lumbar spine (LS) or femoral neck (FN), or had a prior major osteoporotic fracture and a T-score [le]-1.0 and [ge]-3.5 at the LS or FN; and had at least two lumbar vertebrae, one femur, and one forearm evaluable by DXA. Subjects received daily calcium ([ge]1000 mg) and vitamin D ([ge]800 IU) supplementation. The primary endpoint was to assess the percent change from baseline in LS BMD at 12 months. Secondary endpoints included the percent change from baseline in total hip (TH), FN, trochanter (TR), and forearm (1/3R) BMD at month 12. Sensitivity analyses for LS BMD were done by controlling for baseline covariates (testosterone levels, BMD T-scores, race, geographic region and 10-year major osteoporotic fracture risk). The efficacy of DMAb on LS BMD was assessed in different subgroup populations (grouped by testosterone levels, minimum BMD T-scores, and 10-year major osteoporotic fracture risk). Safety analysis included all randomized subjects who received [ge]1 dose of investigational product.[br][bold]Results: [/bold]A total of 242 subjects were randomized; 228 (94.2%) completed one year of treatment. The mean (SD) age was 65 (9.8) yrs and 15% of subjects had testosterone concentration [lt]250 mg/dL at baseline. After 12 months of treatment with DMAb, BMD increased from baseline by 5.7%, 2.4%, 2.1%, 3.1%, and 0.6% at the LS, TH, FN, TR, and 1/3R, respectively (all [italic]p[/italic][lt]0.01 compared with placebo and adjusted for multiplicity). Sensitivity analyses for LS BMD were consistent with the primary analysis (mean differences from placebo were significant, [italic]p[/italic][lt]0.01), indicating the primary analysis results were robust. DMAb treatment resulted in significant increases in LS BMD in all subgroups (mean differences from placebo were significant, [italic]p[/italic][lt]0.005), indicating DMAb is effective across different clinical situations. Adverse event incidence was similar between groups.[br][bold]Conclusions: [/bold]One year of DMAb therapy in men with low BMD was well tolerated and resulted in significant increases in BMD at all measured skeletal sites.[br][br]Disclosures: EO: Research Funding, Amgen, Eli Lilly & Company, Merck & Co.; Consultant, Eli Lilly & Company, Merck & Co., Amgen, Wright Medical Technology. CST: Coinvestigator, Amgen. BLL: Research Funding, Amgen, Eli Lilly & Company, Novartis Pharmaceuticals, MSD; Consultant, Amgen, Eli Lilly & Company, Novartis Pharmaceuticals, Servie, Nycomed, MSD. RC: Advisory Group Member, Amgen, Eli Lilly & Company, Servier, Merck & Co., Novartis Pharmaceuticals; Speaker, Amgen, Ipsen, Servier, Roche Pharmaceuticals. EC: Advisory Group Member, Amgen, Eli Lilly & Company, Servier, Merck & Co., Novartis Pharmaceuticals; Speaker, Amgen, Ipsen, Servier, Roche Pharmaceuticals. DLK: Coinvestigator, Eli Lilly & Company, GlaxoSmithKline, Roche Pharmaceuticals; Advisory Group Member, Merck & Co., Novartis Pharmaceuticals, Amgen, Pfizer Global R&D. J-YR: Advisory Group Member, Servier, Novartis Pharmaceuticals, Negma, Lilly USA, LLC, Wyeth Pharmaceuticals, Genzyme Corporation, Roche Diagnostics, Merck & Co., Nycomed, NPS, Theramex, UCB; Speaker, Merck Sharp and Dohme, Lilly USA, LLC, Rottapharm, IBSA, Genevrier, Novartis Pharmaceuticals, Roche Pharmaceuticals, GlaxoSmithKline, Teijin, Teva, Ebewee Pharma, Zodiac, Analis, Theramex, Nycomed, Novo Nordisk; Investigator, Bristol Myers Squibb, Merck Sharp and Dhome, Rottapharm, Teva, Lilly, Novartis, Roche, GlaxoSmithKline, Amgen, Servier. AK: Coinvestigator, Amgen. EML: Principal Investigator, Amgen, Eli Lilly & Company, Merck & Co., Novartis Pharmaceuticals, GlaxoSmithKline, Warner Chilcott Pharmaceuticals; Speaker Bureau Member, Amgen, Eli Lilly & Company, Novartis Pharmaceuticals, Warner Chilcott Pharmaceuticals; Scientific Board Member, Amgen, Eli Lilly & Company, Merck & Co., Novartis Pharmaceuticals; Consultant, GlaxoSmithKline. PDM: Research Funding, Amgen, Eli Lilly & Company, Genentech, Inc., Merck & Co., Novartis Pharmaceuticals, Warner Chilcott Pharmaceuticals; Speaker Bureau Member, Amgen, Eli Lilly & Company, Genentech, Inc., Merck & Co., Novartis Pharmaceuticals, Warner Chilcott Pharmaceuticals, GlaxoSmithKline. MAB: Research Funding, Amgen, Eli Lilly & Company, Merck & Co.; Speaker Bureau Member, Amgen, Eli Lilly & Company, Warner Chilcott Pharmaceuticals. MRM: Consultant, Merck & Co. HGB: Consultant, Amgen, Merck & Co., Takeda, Tarsa; Investigator, Amgen, Merck & Co., Nordic Bioscience A/S, Novartis Pharmaceuticals, Takeda, Tarsa; Speaker Bureau Member, Amgen. BA: Research Funding, Amgen, Novartis Pharmaceuticals; Consultant, Amgen, Novartis Pharmaceuticals, Eli Lilly & Company, Nycomed; Speaker Bureau Member, Merck & Co., Nycomed. Y-CY: Employee, Amgen. RWW: Employee, Amgen. SS: Employee, Amgen. AG: Employee, Amgen. JWH: Employee, Amgen. SB: Research Funding, Amgen; Consultant, Amgen. Nothing to Disclose: UG, OL 2012-06-23T12:15:00 Theater A 2012-06-23T00:00:00 1899-12-30T12:15:00 1789 41 102 OR08-5 OR05-01 Saturday 47 2012


48 ENDO12L_OR08-6 ORAL SESSION: Osteoporosis Physiology [amp] Clinical Trials (11:15 AM-12:45 PM) Behavior of Bone Turnover Markers in Infertile Women Undergoing Follicle-Stimulating Hormone Administration for [italic]In Vitro[/italic] Fertilization Procedures Umberto Omodei, Gherardo Mazziotti, Gloria Donarini, Monica Gola, Valentina Guella, Franca Pagani, Teresa Porcelli, Andrea Giustina Universit[agrave] degli Studi di Brescia, Brescia, Italy; Universit[agrave] degli Studi di Brescia, Brescia, Italy; Fondazione Poliambulanza, Brescia, Italy There is experimental but limited clinical evidence to suggest that the increase in follicle-stimulating hormone (FSH) secretion, as it occurs since the first phase of perimenopause, may be involved in determining post-menopausal bone loss. In this prospective study, we evaluated for the first time the sequential profile of serum bone turnover markers during acute stimulation with recombinant FSH in 29 infertile women (age ranging from 30 and 40 years) undergoing in vitro fertilization procedures. In this clinical model, the skeletal effects of exogenous FSH were evaluated after pharmacological suppression of endogenous gonadotropin and estrogen production. The women were evaluated for serum osteocalcin (Elecsys N-MID Osteocalcina, Roche Diagnostics, Monza, Italy; reference range: 11-32 [mu]g/L), [beta]-CTX (Elecsys [beta]-CrossLaps Roche Diagnostics, Monza, Italy; reference range: 0.10-0.62 [mu]g/L), FSH and estradiol at baseline (T0), 12-20 days after the first dose of GnRH analog (Leuprolide 1 mg subcutaneously each day) (T1), 3 days (T2), 10 days (T3) after recombinant FSH administration (Meropur 75 in the morning and Gonal-F 1050 in the evening at doses between 75 and 225 UI each day) and at pick-up (T4) and at the embryo-transfer day (T5). All women were taking also prednisolone between T4 and T5. At study entry, all patients had serum osteocalcin and [beta]-CTX values in the reference ranges for pre-menopausal women. At T1, the suppression of FSH and E2 secretion, as affect of GnRH analog administration, was accompanied by a significant increase in serum [beta]-CTX values as compared to T0 (p[lt]0.001), whereas no significant change in serum osteocalcin was observed at this time. After the administration of recombinant FSH, a rapid increase in serum FSH was observed, whereas serum E2 values increased more slowly. At this time, the increase in serum FSH values (with E2 in normal range) was not associated with any significant change in median serum [beta]-CTX and osteocalcin values as compared to T1. At T3 (when both FSH and E2 were high), serum [beta]-CTX values decreased significantly as compared to T1 (p[lt]0.001), whereas osteocalcin remained unchanged. At T4, serum [beta]-CTX and osteocalcin remained unchanged as compared to T3, whereas both markers decreased significantly (p[lt]0.001 for both parameters) at T5. In conclusion, our model suggests that FSH, at high-normal circulating values, does not seem to acutely exert relevant direct effects on bone.[br][br]Nothing to Disclose: UO, GM, GD, MG, VG, FP, TP, AG 2012-06-23T12:30:00 Theater A 2012-06-23T00:00:00 1899-12-30T12:30:00 2195 41 103 OR08-6 OR05-01 Saturday 48 2012


49 ENDO12L_OR09-1 ORAL SESSION: Pediatric Endocrinology, General (11:15 AM-12:45 PM) Mutations in the [italic]AAAS[/italic] Gene in 215 Patients with Triple A Syndrome [mdash] Analysis of a Genotype/Phenotype Relationship Angela Huebner, Daniela Klaus, Dana Landgraf, Petra Mitzscherling, Katrin Koehler Technical University Dresden, Dresden, Germany The triple A syndrome (MIM*231550) is a rare autosomal recessive disorder characterized by adrenocorticotropic hormone (ACTH) resistant adrenal failure, alacrima, achalasia and a variety of neurological and dermatological features. The syndrome is caused by mutations in the [italic]AAAS[/italic] gene encoding ALADIN, a protein of the nuclear pore complex. We investigated N=173 families including 215 patients with clinically suggested triple A syndrome. In 142 families we identified 62 different [italic]AAAS[/italic] mutations including 19 nonsense mutations, 15 deletions and 2 insertions leading to a frameshift and a truncated protein, 13 splice defects and 13 missense mutations which were present in a homozygous or compound heterozygous form. Although the mutations were scattered over the entire gene a few mutation hotspots became apparent including c.43C[gt]A (exon1), c.787T[gt]C (exon8) and c.1331+1G[gt]A (intron 14), each occurring in 20 and more families from different regions. In 31 families with triple A syndrome we could not detect [italic]AAAS[/italic] mutations indicating genetic heterogeneity. Two patients carry [italic]AAAS[/italic] mutations in a heterozygous form. The phenotype of patients who either lack a mutation or appear to be heterozygous does not differ from patients with a proven [italic]AAAS[/italic] mutation.[br]Genotype/phenotype analyses revealed a highly variable occurrence, age of onset and severity of all clinical symptoms between patients with the same [italic]AAAS[/italic] mutation. This is reflected by the considerable intrafamilial variability which may also concern the three main symptoms. Whereas alacrima appears to be the earliest and most consistent symptom (93 % of all patients), achalasia was observed in 86 %. Adrenal insufficiency (75 % of all patients) usually occurs in the first decade of life but may occur only in the second decade or even later. The severity and progression of neurological features resembling those of a mixed hereditary motor and sensory neuropathy with axonal degeneration cannot be correlated to the localization and the nature of the mutations in the [italic]AAAS[/italic] gene. Nasal speech and a marked generalized hyperreflexia appear to be pathognomonic signs.In conclusion, in view of the highly variable clinical course, the triple A syndrome seems to be an underdiagnosed disorder. [italic]AAAS[/italic] mutation screening should therefore be extended to patients who exhibit only single features of the disease. The obvious lack of a genotype/phenotype relationship is suggestive of modifying genes/factors which have to be determined.[br][br]Sources of Research Support: German Research Foundation HU/895 3-3,3-4,3-5 and 4-1 awarded to AH.[br][br]Nothing to Disclose: AH, DK, DL, PM, KK 2012-06-23T11:15:00 Theater C 2012-06-23T00:00:00 1899-12-30T11:15:00 1257 42 104 OR09-1 OR30-01 Saturday 49 2012


50 ENDO12L_OR09-2 ORAL SESSION: Pediatric Endocrinology, General (11:15 AM-12:45 PM) Finite Element and Extended Cortical Analysis in Young Amenorrheic Athletes, Eumenorrheic Athletes and Non-Athletes Kathryn E Ackerman, Melissa Putman, Gabriela Guereca, Alexander P Taylor, Lisa Pierce, David B Herzog, Anne Klibanski, Mary Bouxsein, Madhusmita Misra Harvard Medical School, Boston, MA; Children[apos]s Hospital Boston, Boston, MA; Massachusetts General Hospital, Boston, MA; Massachusetts General Hospital, Boston, MA; Massachusetts General Hospital for Children, Boston, MA CONTEXT: Lower bone mineral density (BMD) in young amenorrheic athletes (AA) compared to eumenorrheic athletes (EA) and non-athletes may increase fracture risk during a critical time of bone accrual. Finite element analysis (FEA) is a unique [italic]in vivo[/italic] tool to estimate bone strength, while extended cortical analysis (ECA) allows assessment of cortical parameters that may contribute to bone strength using data from high-resolution peripheral QCT (HRpQCT).[br]OBJECTIVE: We hypothesized that stiffness and failure load are impaired in AA at the distal radius and tibia compared to EA and non-athletes despite weight-bearing exercise.[br]DESIGN AND SETTING: Cross-sectional study at the Clinical Research Center[br]SUBJECTS: Subjects included 34 female endurance athletes 14-21 years old involved in weight-bearing sports (17 AA, 17 EA) and 16 non-athletes of normal-weight (BMI 10th-90th percentiles) and comparable age and maturity. All subjects had body composition assessment using dual energy x-ray absorptiometry (DXA).[br]OUTCOME MEASURES: We used ECA to assess cortical parameters including porosity, and FEA to assess stiffness and failure load, using images obtained from HRpQCT.[br]RESULTS: At the weight-bearing tibia, both groups of athletes had greater cortical perimeter and porosity (p=0.02 and 0.01 respectively), and greater trabecular area than non-athletes (p=0.01), while percent cortical area was lowest in AA (14.6[plusmn]3.2% vs. 16.4[plusmn]3.0% in EA and 18.6[plusmn]4.9% in non-athletes; p=0.01). Despite greater cortical porosity in EA than non-athletes, stiffness and failure load were higher in EA (p=0.01 for both). This advantage, however, was lost in AA. At the non-weight-bearing radius, failure load and stiffness were lower in AA than non-athletes [failure load (kN): 3.60 [plusmn] 0.77 in AA, 4.05 [plusmn] 0.60 in EA and 4.20 [plusmn] 0.74 in non-athletes, p=0.048; stiffness (kN/m): 70.5[plusmn]14.8 in AA, 79.5[plusmn]12.1 in EA and 83.0[plusmn]15.6 in non-athletes, p=0.04]. After controlling for lean mass and menarchal age, athletic status accounted for 5-9% of the variability in stiffness and failure load, while menarchal age accounted for 8-23%, and lean mass for 12-37%.[br]CONCLUSION: AA have decreased bone strength at the distal radius (non-weight-bearing site) compared with non-athletes, and lose the advantage of weight-bearing exercise seen in EA at the distal tibia.[br][br]Sources of Research Support: NIH grants 1 UL1 RR025758-01 and 1 R01 HD060827-01A1.[br][br]Nothing to Disclose: KEA, MP, GG, APT, LP, DBH, AK, MB, MM 2012-06-23T11:30:00 Theater C 2012-06-23T00:00:00 1899-12-30T11:30:00 51 42 105 OR09-2 OR30-01 Saturday 50 2012


51 ENDO12L_OR09-3 ORAL SESSION: Pediatric Endocrinology, General (11:15 AM-12:45 PM) Inhibition of Pref-1 by Estradiol in Adolescent Girls with Anorexia Nervosa Is Associated with Improvement in Lumbar Bone Mineral Density Alexander T Faje, Pouneh Fazeli, Debra K Katzman, Karen K Miller, Anne Breggia, Clifford J Rosen, Nara Mendes, Madhusmita Misra, Anne Klibanski Massachusetts General Hospital and Harvard Medical School, Boston, MA; Hospital for Sick Children and University of Toronto, Toronto, Canada; Maine Medical Center Research Institute, Portland, ME; Massachusetts General Hospital for Children and Harvard Medical School, Boston, MA Regulation of mesenchymal stem cell (MSC) differentiation into the osteoblast or the adipocyte lineage may influence bone formation. Adolescents with anorexia nervosa (AN) have low bone mineral density (BMD) and decreased bone formation. Preadipocyte factor 1 (Pref-1), an inhibitor of adipocyte and osteoblast differentiation, is elevated in states of estrogen deficiency and may negatively affect BMD in adolescent girls with AN.[br][bold]Objective:[/bold] To (i) compare levels of Pref-1 in adolescent girls with AN and controls, (ii) investigate the effects of transdermal estradiol on Pref-1 in AN, and (iii) examine associations of changes in Pref-1 with changes in lumbar BMD and markers of bone turnover.[br][bold]Design and Methods:[/bold] Sixty-nine girls (44 with AN and 25 normal-weight controls) 13-18 years of age were studied at baseline. Subjects were selected from a prior study based on their lumbar spine BMD Z-scores. The subset included all subjects with AN who had a baseline lumbar BMD Z-score less than -0.5 and normal-weight controls with lumbar BMD Z-scores between +1.0 and -1.0. Twenty-two AN girls with a bone age of [ge] 15 years were randomized to 100mcg transdermal estradiol (n = 13) or placebo (n = 9) in a double-blind study for 12 months. Subjects randomized to transdermal estradiol received medroxyprogesterone 2.5mg daily for 10 days each month. All subjects also received calcium carbonate (1200mg) and vitamin D (400 IU) daily.[br][bold]Results:[/bold] Although Pref-1 levels did not differ in AN versus controls (0.246 [plusmn] 0.015 vs. 0.267 [plusmn] 0.022 ng/ml, p = 0.46) at baseline, levels significantly decreased in girls with AN after treatment with transdermal estradiol compared with placebo (-0.015 [plusmn] 0.016 vs. 0.060 [plusmn] 0.026 ng/ml, p = 0.02). Changes in Pref-1 over twelve months correlated inversely with changes in lumbar BMD (r = -0.48, p = 0.02) and positively with changes in CTX (r = 0.73, p = 0.01). Levels of Pref-1 were inversely associated with estradiol levels in subjects with AN (r = -0.67, p = 0.004) after 12 months of treatment with transdermal estradiol or placebo.[br][bold]Conclusions:[/bold] One year of estrogen replacement suppresses Pref-1 in girls with AN. Additionally, Pref-1 levels correlate inversely with changes in BMD and positively with changes in CTX, a marker of bone resorption. Inhibition of Pref-1 may in part mediate the beneficial effects of transdermal estradiol replacement on bone mass in adolescent girls with AN.[br][br]Nothing to Disclose: ATF, PF, DKK, KKM, AB, CJR, NM, MM, AK 2012-06-23T11:45:00 Theater C 2012-06-23T00:00:00 1899-12-30T11:45:00 87 42 106 OR09-3 OR30-01 Saturday 51 2012


52 ENDO12L_OR09-4 ORAL SESSION: Pediatric Endocrinology, General (11:15 AM-12:45 PM) Effect of Oxandrolone Versus Placebo on Body Composition and Bone Density in Klinefelter Syndrome: Results of a Two-Year, Double-Blind, Placebo-Controlled Trial Martha Zeger Bardsley, Karen Kowal, Judith Levine Ross Thomas Jefferson University/Nemours, Philadelphia, PA [bold]Introduction:[/bold] Klinefelter[apos]s syndrome (KS) is a relatively common genetic disorder defined by the karyotype 47,XXY. Males with KS often have decreased muscle tone, increased central adiposity, and are at increased risk for metabolic syndrome. The KS phenotype may be the result of androgen deficiency in utero, infancy and childhood. Androgen replacement is standard in adolescent and adult KS males but has not been used earlier in childhood. The results presented are from a randomized clinical trial to study the effects of childhood androgen replacement on the KS neurocognitive and physical phenotypes.[br][bold]Objective:[/bold] To determine the effects of androgen treatment for two years in boys with KS on body composition, including muscle mass, body fat, and bone density.[br][bold]Study design:[/bold] This double-blind, placebo-controlled clinical trial (2005-2011, NCT00348946) randomized 93 boys, ages 4-12 yrs, to 2 groups: oxandrolone (Ox)(N=46) and placebo (Pl)(N=47). Protocol specified Ox dose was 0.06 mg/kg daily. Study visits occurred every 6 months for 2 years. Measurements included limb circumference, body fat percent by skin fold, BMI and weight. All measurements were converted to standard deviation scores. Bone mass was determined by use of the bone health index (BHI), using automated radiogrammetry (1). Statistical analysis included repeated measures ANCOVA and Fishers Exact test.[br][bold]Results:[/bold] 93 patients enrolled, and the 80 (86%) subjects who completed the 2 year study were compared. Their mean ages at 2 years were (Ox) 9.1[plusmn]2.1 years, n=39 vs. (Pl) 10.2 [plusmn]2.6 years, n=41, (P=0.03). Weight SDS increased in the Ox (0.3[plusmn]0.6) vs. Pl (-0.07[plusmn]0.4) (P[lt].05), but the change in BMI SDS did not differ (P=0.7). Fewer boys had body fat [gt] 25% in the Ox (23%) vs. Pl (49%) group (P=0.03). Increased muscle mass was demonstrated by increased lower leg circumference SDS in Ox group (change from baseline: 0.5[plusmn]0.7 Ox vs. 0.1[plusmn]0.5 Pl, P[lt]0.01). Mean BHI SDS change from baseline was 0.4[plusmn]0.7 (Ox) vs. -0.2 [plusmn]0.6 (Pl) (P [lt]0.01), indicating increased bone mass (cortical thickness) in the Ox group.[br][bold]Conclusions: [/bold]This unique double-blind, randomized clinical trial demonstrates that Ox increases weight SDS but not BMI SDS, and is associated with increased muscle mass, lower percent body fat, and increased bone cortical thickness. Therefore, gradual age-appropriate androgen replacement should be considered to optimize body composition in KS.[br][br](1)Thodberg HH, van Rign RR, Tanaka T, Martin DD, Kreiborg S, A paediatric bone index derived by automated radiogrammetry. Osteoporosis International 2009.[br][br]Sources of Research Support: This study was sponsored by NIH: Study #R01NS050597.[br][br]Disclosures: JLR: Scientific Board Member, Eli Lilly & Company; Speaker, Eli Lilly & Company. Nothing to Disclose: MZB, KK 2012-06-23T12:00:00 Theater C 2012-06-23T00:00:00 1899-12-30T12:00:00 1991 42 107 OR09-4 OR30-01 Saturday 52 2012


53 ENDO12L_OR09-5 ORAL SESSION: Pediatric Endocrinology, General (11:15 AM-12:45 PM) Optimum Macronutrient Content of the Diet for Adolescents with Pre-Diabetes; RESIST a Randomized Control Trial ACTRN12608000416392 Sarah P Garnett, Megan Dunkley, Mandy Ho, Louise A Baur, Manny Noakes, Kate Steinbeck, Helen J Woodhead, Kerryn Chisholm, Susie Burrell, Carolyn R Broderick, Robert Parker, Sukanya De, Shubha Shrinivasan, Geoffrey R Ambler, Michael R Kohn, Chris T Cowell The Children[apos]s Hospital at Westmead, Sydney, Australia; The Children[apos]s Hospital at Westmead, Sydney, Australia; University of Sydney, Sydney, Australia; CSIRO, Adelaide, Australia; Sydney Medical School, University of Sydney, Sydney, Australia; Sydney Children[apos]s Hospital at Campbelltown, Sydney, Australia; The Children[apos]s Hospital at Westmead, Sydney, Australia; The Children[apos]s Hospital at Westmead, Sydney, Australia; The Children[apos]s Hospital at Westmead, Sydney, Australia Rationale: Pre-diabetes and clinical insulin resistance in adolescents are rapidly emerging clinical problems with serious health outcomes. With appropriate management these conditions are potentially reversible. Yet there are no randomised trials of lifestyle interventions in this group to inform clinical practice.[br]Aim: To determine the efficacy and effectiveness of two structured lifestyle interventions, differing only in diet composition, on insulin sensitivity in obese adolescents with pre-diabetes and/or clinical features of insulin resistance.[br]Design: A 12 month RCT taking place at two tertiary hospitals in Sydney, Australia. At baseline adolescents are prescribed metformin and randomised to either a high carbohydrate low fat diet or a moderate carbohydrate increased protein diet. The program commences with an intensive 3 month dietary intervention, followed by a 3 month intensive exercise intervention and a 6 month maintenance phase (1).[br]Inclusion criteria: 10 to 17 year olds who are overweight or obese, with either pre- diabetes and/or clinical features of insulin resistance.[br]Clinical endpoints: Insulin sensitivity index (ISI) (2), adiposity, cardiometabolic profile and fitness. Differences in outcome measures overtime and between trial arms will be examined at 3, 6, 12 and 24 months.[br]Preliminary data: Recruitment is complete, n=111 (66 girls) and 101 have completed 3 months (4 dropped out and the remaining 6 will complete in February 2012). At baseline, most had a family history of obesity (87%), type 2 diabetes (75%) and/or a high risk ethnic background (58%) including Aboriginal, Maori and Pacific Islander. 94 (85%) had acanthosis nigricans and 14 (13%) had pre-diabetes. The mean BMI z-score was 2.3[plusmn]0.3, the median ISI was 1.3 [0.3 to 3.4 and 58 (52%) had SBP and/or DPB [ge] 90th centile, 56 (50%) had ALT and/or GTT [ge] 30 U/L and 70 (63%) had HDL cholesterol [le]1.03mmol/L and/or triglycerides [ge]1.7mmol/L.[br]Discussion: We have recruited an obese population with clinical and biochemical abnormalities placing them at high risk of developing type 2 diabetes. The three month outcomes including differences between dietary groups will be presented.[br][br](1)Garnett SP et al., BMC Public Health 2010; 10:575. (2)Matsuda M et al., Diabetes Care 1999; 22:1462.[br][br]Sources of Research Support: Diabetes Australia Research Trust 2008, BUPA Health, Foundation Australia Pty Limited (formerly MBF Foundation)2008 to 2012; Heart Foundation (#G08S3758)2009 to 2010. SG was supported by a National Health and Medical Research Council Australian Clinical, Research Fellowship(#457225)2007 to 2010 and is supported by a Cancer Institute NSW ECR Fellowship 2011 to 2013.[br][br]Nothing to Disclose: SPG, MD, MH, LAB, MN, KS, HJW, KC, SB, CRB, RP, SD, SS, GRA, MRK , CTC 2012-06-23T12:15:00 Theater C 2012-06-23T00:00:00 1899-12-30T12:15:00 729 42 108 OR09-5 OR30-01 Saturday 53 2012


54 ENDO12L_OR09-6 ORAL SESSION: Pediatric Endocrinology, General (11:15 AM-12:45 PM) Metabolic Syndrome Features in Children Born Post-Term Ahila Ayyavoo, Paul Hofman, Jose Derraik, Sarah Mathai, Peter Stone, Lynn Sadler, Wayne Cutfield University of Auckland, Auckland, New Zealand; National Research Centre for Growth and Development, Auckland, New Zealand; Auckland District Health Board, Auckland, New Zealand [bold]Introduction [/bold][br]We have recently shown from a large Swedish cohort that nearly half of the boys born post-term were overweight or obese at 16 years of age. In contrast, post-term girls were leaner at birth, but had similar weights to control girls at 16 years (Beltrand et al., J Pediatr 2011).[br][bold]Hypothesis[/bold][br]Pre-pubertal children born post-term have features of the metabolic syndrome.[br][bold]Methods[/bold][br]Two groups of healthy pre-pubertal children aged 4[ndash]11 years were studied: post-terms (n=36; born at [ge] 42 weeks gestation) and controls (n=56; 38-40 weeks). Following an overnight fast, we performed a frequently sampled intravenous glucose tolerance test with insulin. Insulin sensitivity (Si) was calculated using Bergman[apos]s minimal model. Biochemical markers of metabolic syndrome were examined. Auxological and DEXA-derived body composition data were obtained. Children also underwent 24-hour blood pressure (BP) monitoring. Data were analysed separately for boys and girls using linear mixed models, controlling for appropriate confounders. Data are expressed as mean[plusmn]SEM.[br][bold]Results[/bold][br][bold]Si[/bold] was reduced in both post-term boys (n=18; 9.7[plusmn]1.21 vs 14.2[plusmn]1.1x10[sup]-4[/sup] min[sup][minus]1[/sup][middot](mU/L); p=0.014) and girls (n=18; 7.3[plusmn]0.7 vs 10.4[plusmn]1.2 x 10[sup]-4[/sup] min[sup][minus]1[/sup][middot](mU/L)); p=0.046) compared to controls. There was an appropriate compensatory increase in [bold]acute insulin response[/bold] in post-term boys (549 [plusmn]101 vs 269[plusmn]25 mU/L; p=0.009), but not in post-term girls (378.5[plusmn]53.3 vs 277[plusmn]29.8 mU/L; p=0.65). [bold]IGFBP1[/bold] was lower in post-term boys (9.9ng/ml[plusmn]1.57 vs 18.31[plusmn]2.14 ng/ml; p=0.027) and girls (9.23[plusmn]1.47 vs 17.44[plusmn]2.14 ng/ml; p=0.08).[br]There were further changes suggestive of metabolic syndrome in post-term boys, including elevated [bold]IGFBP3[/bold] (3815 ng/ml [plusmn]241: 3313[plusmn]110 ng/ml; p=0.027), and reductions in both [bold]nocturnal systolic [/bold](8.2[plusmn]1.0 vs 13.8[plusmn]1.0 mmHg; p[lt]0.0001) and [bold]nocturnal diastolic [/bold](14.9[plusmn]0.9 vs 19.8[plusmn]1.3 mmHg; p 0.004) [bold]BP dip[/bold]. Post-term boys were also dyslipidemic, with higher [bold]total cholesterol[/bold] (4.4 [plusmn]0.3 vs 3.9[plusmn]0.1 mmol/l; p=0.058), [bold]LDL[/bold] (2.6[pusmn]0.2 vs 2.2[plusmn]0.1 mmol/l; p=0.069) and [bold]triglycerides[/bold] (0.84[plusmn]0.07 vs 0.7[plusmn]0.05 mmol/l; p=0.088) than boys born at term. Further, post-term boys had higher serum [bold]leptin[/bold] concentrations (5.9 [plusmn]1.4 vs 3.2[plusmn]0.4 ng/ml; p=0.056) despite similar body composition as boys born at term (21.8[plusmn]2.4 vs 17.5[plusmn]1.1 %; p=0.47).[br][bold]Conclusion[/bold]s[br]Post-term boys and girls have insulin resistance. Post-term boys have additional features of metabolic syndrome. We speculate that insulin resistance precedes obesity in post-term children.[br][br]Sources of Research Support: National Research Centre for Growth and Development; APEG Research Grants 2010 [amp] 2011.[br][br]Nothing to Disclose: AA, PH, JD, SM, PS, LS, WC 2012-06-23T12:30:00 Theater C 2012-06-23T00:00:00 1899-12-30T12:30:00 518 42 109 OR09-6 OR30-01 Saturday 54 2012


55 ENDO12L_S18-1 ORAL SESSION: Clinical Trial Symposium (3:45 PM-5:15 PM) Weight Loss, Exercise, or Both and Cardiometabolic Risk Factors in Obese Older Adults: Results of a Randomized Controlled Trial Matthew F Bouchonville, Krupa Shah, Reina Armamento-Villareal, David R Sinacore, Clifford Qualls, Dennis T Villareal University of New Mexico School of Medicine, Albuquerque, NM; University of Rochester Medical Center, Rochester, NY; Washington University School of Medicine, St Louis, MO; New Mexico Veterans Affairs Health Care System, Albuquerque, NM; University of New Mexico School of Medicine, Albuquerque, NM; Washington University School of Medicine, St Louis, MO; Washington University School of Medicine, St Louis, MO; New Mexico Veterans Affairs Health Care System, Albuquerque, NM; University of New Mexico School of Medicine, Albuquerque, NM [bold]Background: [/bold]In older adults, obesity exacerbates the age-related decline in physical and cardiometabolic health; however, the appropriate treatment for obese older adults is controversial. We recently reported that the combination of weight loss and exercise provides greater improvement in physical function than either intervention alone ([italic]N Engl J Med[/italic] 2011). We now report the results of the independent and combined effects of weight loss and exercise on cardiometabolic risk factors in this population.[br][bold]Methods: [/bold]In this 1-year randomized controlled trial, 107 obese (BMI [ge] 30 mg/kg[sup]2[/sup]) older (age [ge] 65 years) adults were randomly assigned to control, diet, exercise, and combined diet-exercise. The primary outcome was the change in insulin sensitivity index (ISI), measured from the oral glucose tolerance test (OGTT). Secondary outcomes included glucose and insulin response to the OGTT, waist circumference and abdominal adiposity, systolic and diastolic blood pressure (BP), and serum concentration of lipids and adipocytokines.[br][bold]Results: [/bold]Ninety-three participants (87%) completed the trial. In the intention to treat analyses, the ISI improved in the diet group (70%) and in the diet-exercise group (86%), but not in the exercise group or the control group (between-group [italic]P[/italic][lt]0.05). Accordingly, both the glucose and the insulin area under the curves during the OGTT improved in the diet group (10%) and in the diet-exercise group (15-20%), but not in the exercise or the control group (between-group [italic]P[/italic][lt]0.05). Moreover, systolic BP (-9% and -10%), diastolic BP (-8% and -6%), waist circumference (-7% and -7%), abdominal visceral fat (-25% and -33%), serum triglycerides (-26% and -27%), C-reactive protein (-27% and -27%), and TNF-R (-7% and -9%) decreased while adiponectin (21% and 33%) increased in the diet group and in the diet-exercise group, but not in the exercise group or the control group (all between-group P[lt]0.05). Body weight decreased in the diet group (10%) and in the diet-exercise group (9%) but did not decrease in the exercise group or the control group (between-group [italic]P[/italic][lt]0.05).[br][bold]Conclusions: [/bold]Diet-induced weight loss but not exercise training improves insulin sensitivity and multiple other cardiometabolic risk factors simultaneously in obese older adults. Therefore, in addition to additive effects of weight loss with exercise on physical function, weight loss is particularly important in obese older adults to decrease the risk for cardiometabolic syndrome.[br][br]Villareal DT et al., N Engl J Med 2011; 364:1218-29.[br][br]Sources of Research Support: RO1 Grant; National Institute of Aging; NIH Grant AG025501 awarted to DTV.[br][br]Nothing to Disclose: MFB, KS, RA-V, DRS, CQ, DTV 2012-06-23T15:45:00 362 2012-06-23T00:00:00 1899-12-30T15:45:00 2288 101 896 S18-1 OR300-01 Saturday 55 2012


56 ENDO12L_S18-2 ORAL SESSION: Clinical Trial Symposium (3:45 PM-5:15 PM) The Effect of Six Years of Denosumab Treatment on New Vertebral and Nonvertebral Fractures in Postmenopausal Women with Osteoporosis: Results from the FREEDOM Extension Trial Henry G Bone, Jacques P Brown, Roland Chapurlat, Nathalie Franchimont, Maria L Brandi, Edward Czerwinski, Marc-Antoine Krieg, Zulema Man, Dan Mellstrom, Sebastiao C Radominski, Jean Yves Reginster, Heinrich Resch, Jose A Roman, Nadia S Daizadeh, Andrea Wang, Michelle L Geller, Rachel B Wagman, Socrates Papapoulos Michigan Bone and Mineral Clinic, Detroit, MI; Laval University and CHUQ Research Centre, Quebec City, Canada; Universit[eacute] de Lyon, Lyon, France; Amgen Inc, Thousand Oaks, CA; University of Florence, Florence, Italy; Krakow Medical Center, Krakow, Poland; University Hospital of Lausanne, Lausanne, Switzerland; Centro TIEMPO, Buenos Aires, Argentina; Sahlgrenska University Hospital, G[ouml]teborg, Sweden; Universidade Federal do Paran[aacute], Curitiba, Brazil; University of Li[egrave]ge, Li[egrave]ge, Belgium; St Vincent Hospital, Vienna, Austria; Hospital Universitario La Fe, Valencia, Spain; Amgen Inc, Thousand Oaks, CA; Amgen Inc, Thousand Oaks, CA ; Leiden University Medical Center, Leiden, Netherlands The FREEDOM trial open-label extension is evaluating the long-term safety and efficacy of denosumab (DMAb) for up to 10 years. We report the intention-to-treat (ITT) results for women who participated in the first 3 years of the extension, representing up to 6 years of DMAb exposure. We also report the results for the pre-specified per protocol (PP) and modified per protocol (MPP) subsets.[br]During the extension, each woman is scheduled to receive 60 mg DMAb every 6 months and supplemental calcium and vitamin D daily. For the analyses here, women from the FREEDOM placebo group received 3 years of DMAb (cross-over group) and women from the FREEDOM DMAb group received 3 more years of DMAb for a total of 6 years (long-term group). The PP and MPP subsets excluded subjects who were non-compliant with the protocol and the MPP subset further excluded subjects who missed [ge] 2 doses of DMAb during the extension.[br]Of the 5928 women eligible for the extension, 4550 (77%) enrolled (N = 2207 cross-over; N = 2343 long-term). During the first 3 years of DMAb treatment during the extension, the cross-over group had significant gains in bone mineral density (BMD) at the lumbar spine and total hip, and further significant increases in BMD occurred over years 4 to 6 in the long-term group. Serum CTX was rapidly and similarly reduced after the 1[sup]st[/sup] (cross-over) or 7[sup]th[/sup] (long-term) DMAb dose with the characteristic attenuation observed at the end of the dosing period.[br]In FREEDOM, DMAb reduced the risk of new vertebral (2.3% DMAb vs 7.2% placebo) and nonvertebral (6.5% DMAb vs 8.0% placebo) fractures over 3 years. In the first 3 years of the extension, for the cross-over group, incidences of new vertebral (2.8%) and nonvertebral (5.6%) fractures were similar to the FREEDOM DMAb group. In the long-term group, fracture incidences remained low (3.5% for new vertebral and 3.8% for nonvertebral fractures). The fracture efficacy results for the PP and MPP subsets were consistent with the ITT analysis. Incidences of adverse events (AEs) and serious AEs did not increase over time with DMAb. No subjects developed neutralizing antibodies to DMAb.[br]DMAb treatment for 3 years in the cross-over group reproduced FREEDOM efficacy observations. DMAb treatment for 6 years (long-term group) remained well tolerated, maintained reduced bone turnover, and continued to significantly increase BMD. Fracture incidence remained low in the ITT, PP, and MPP populations.[br][br]Sources of Research Support: Amgen Inc. sponsored this study.[br][br]Disclosures: HGB: Consultant, Amgen, Merck & Co., Takeda, Tarsa; Investigator, Amgen, Merck & Co., Nordic Bioscience A/S, Novartis Pharmaceuticals, Takeda, Tarsa; Speaker Bureau Member, Amgen. JPB: Investigator, Amgen, Eli Lilly & Company, Merck & Co., Novartis Pharmaceuticals, Warner Chilcott Pharmaceuticals; Advisory Group Member, Amgen, Eli Lilly & Company, Merck & Co., Novartis Pharmaceuticals, Warner Chilcott Pharmaceuticals; Speaker, Amgen, Eli Lilly & Company, Novartis Pharmaceuticals. RC: Advisory Group Member, Amgen, Eli Lilly & Company, Servier, Merck & Co., Novartis Pharmaceuticals; Speaker, Amgen, Ipsen, Servier, Roche Pharmaceuticals. NF: Employee, Amgen. EC: Investigator, Eli Lilly & Company, Novartis Pharmaceuticals, Roche Pharmaceuticals, Amgen, Pfizer, Inc., Servier, Merck Serono S.A., Astra Zeneca, Ardea Biosciences, INC Research, Shire, Biotest AG, Andromeda Biotech Ltd., Johnson & Johnson; Speaker, Amgen, Roche Pharmaceuticals, Servier, Zentiva. SCR: Advisory Group Member, Novartis Pharmaceuticals, Aventis Pharmaceuticals; Study Investigator, Bristol-Myers Squibb, Amgen, Roche Pharmaceuticals, Pfizer, Inc. JYR: Advisory Group Member, Servier, Novartis Pharmaceuticals, Negma, Lilly USA, LLC, Wyeth Pharmaceuticals, Genzyme Corporation, Roche Diagnostics, Merck & Co., Nycomed, NPS, Theramex, UCB; Speaker, Merck Sharp and Dohme, Lilly USA, LLC, Rottapharm , IBSA, Genevrier, Novartis Pharmaceuticals, Roche Pharmaceuticals, GlaxoSmithKline, Teijin, Teva, Ebewee Pharma, Zodiac, Analis, Theramex, Nycomed, Novo Nordisk; Investigator, Bristol Myers Squibb, Merck Sharp and Dhome, Rottapharm, Teva, Lilly, Novartis, Roche, GlaxoSmithKline, Amgen, Servier. NSD: Employee, Amgen. AW: Employee, Amgen. MLG: Employee, Amgen. RBW: Employee, Amgen. SP: Medical Advisory Board Member, Amgen, Eli Lilly & Company, GlaxoSmithKline, Merck & Co., Novartis Pharmaceuticals; Speaker, Amgen, GlaxoSmithKline; Scientific Board Member, Merck & Co. Nothing to Disclose: MLB, M-AK, ZM, DM, HR, JAR 2012-06-23T16:00:00 362 2012-06-23T00:00:00 1899-12-30T16:00:00 1686 101 897 S18-2 OR300-01 Saturday 56 2012


57 ENDO12L_S18-3 ORAL SESSION: Clinical Trial Symposium (3:45 PM-5:15 PM) Efficacy of Recombinant Human Parathyroid Hormone (rhPTH[1[ndash]84]) for the Treatment of Adults with Hypoparathyroidism: A Randomized, Double-Blind, Placebo-Controlled Study John Bilezikian, Leif Mosekilde, Dolores Shoback, Tamara Vokes, Michael Mannstadt, Bart Clarke, Hjalmar Lagast, Roger Garceau Columbia University, College of Physicians [amp] Surgeons, New York, NY; Aarhus University Hospital, Aarhus, Denmark; San Francisco VA Medical Center, University of California, San Francisco, CA; University of Chicago, Chicago, IL; Massachusetts General Hospital and Harvard Medical School, Boston, MA; Mayo Clinic Rochester, Rochester, MN; NPS Pharmaceuticals, Bedminster, NJ Therapy for hypoparathyroidism (HypoPARA) is limited to symptomatic management, often with large amounts of calcium (Ca) and vitamin (Vit) D metabolites leading to concern for possible long-term complications such as end-organ damage. We conducted a randomized, double-blind, placebo (PBO)-controlled study to assess whether replacement therapy with recombinant human parathyroid hormone (rhPTH[1[ndash]84]) reduced Ca and Vit D needs, while normalizing or maintaining albumin-corrected total serum Ca concentration.[br]During a 2[ndash]16 week optimization period, oral Ca and active Vit D doses were adjusted to achieve and maintain serum Ca concentrations of 8.0[ndash]9.0 mg/dL for 2 weeks. Subjects received daily 50 [mu]g subcutaneous injections of rhPTH(1[ndash]84) or PBO for 24 weeks. The rhPTH(1[ndash]84) dose could be increased to 75 or 100 [mu]g if oral Ca and active Vit D doses were sufficiently high to allow for reduction. Regular serum Ca measurements influenced further adjustments in Ca and Vit D requirements in order to maintain serum Ca levels of 8.0[ndash]9.0 mg/dL. A [ldquo]responder[rdquo] was defined as subject whose need for oral Ca and active Vit D could be reduced by 50% at Week 24 while maintaining a serum Ca level of greater than or equal to baseline. Adverse events were monitored throughout treatment and during a 4-week follow-up period.[br]The total number of study subjects was 134 randomized (2:1) to rhPTH(1[ndash]84) or PBO (n=90 and n=44, respectively). A total of 84 subjects in the rhPTH(1[ndash]84) group and 37 in the PBO group completed the study. At the end of treatment, 48/90 (53.3%) subjects in the rhPTH(1[ndash]84) group and 1/44 (2.3%) in the PBO group were responders (treatment difference, 51.1; 95% confidence interval 39.9[ndash]62.3; [italic]P [/italic][lt] 0.001). The mean dose of oral Ca increased from baseline by 5.7% in the PBO group but decreased by 52.0% in the rhPTH(1[ndash]84) group ([italic]P [/italic][lt] 0.001). Active Vit D was reduced by 30.5% in the PBO group and 78.4% in the rhPTH(1[ndash]84) group ([italic]P [/italic][lt] 0.001). At study end, 37 (41.1%) subjects in the rhPTH(1[ndash]84) group and 1 (2.3%) in the PBO group achieved independence from Vit D and could be maintained on an oral Ca dose of [le]500 mg/day. Treatment with rhPTH(1[ndash]84) was generally well tolerated.[br]Treatment of HypoPARA with rhPTH(1[ndash]84) significantly reduces the need for large quantities of Ca and active Vit D, with a majority of responders becoming independent of Vit D and high-dose Ca.[br][br]Sources of Research Support: NPS Pharmaceuticals.[br][br]Disclosures: JB: Advisory Group Member, NPS; Principal Investigator, NPS. DS: Advisory Group Member, NPS; Investigator, NPS. TV: Advisory Group Member, NPS. MM: Advisory Group Member, NPS. BC: Advisory Group Member, NPS. HL: Employee, NPS. RG: Employee, NPS. Nothing to Disclose: LM 2012-06-23T16:15:00 362 2012-06-23T00:00:00 1899-12-30T16:15:00 1682 101 898 S18-3 OR300-01 Saturday 57 2012


58 ENDO12L_S18-4 ORAL SESSION: Clinical Trial Symposium (3:45 PM-5:15 PM) Achievement of Composite of Recommended Goals in Obese Subjects with Type 2 Diabetes Mellitus Using Extended-Release Phentermine/Topiramate in a 56-Week Weight Loss Intervention Donna H Ryan, W Timothy Garvey, Wesley W Day Pennington Biomedical Research Center, Baton Rouge, LA; University of Alabama at Birmingham, Birmingham, AL; Vivus, Inc, Mountain View, CA Current goals for management of type 2 diabetes mellitus (T2DM) include weight loss (WL) and reduction of HbA1c to [lt]6.5% and systolic blood pressure (SBP) to [lt]130 mg/dL. The 56-week, randomised CONQUER study in overweight/obese subjects with [ge]2 weight-related comorbidities showed significant WL with extended-release phentermine/topiramate (PHEN/TPM ER) vs placebo (PBO). This post-hoc analysis assessed percentage of subjects in CONQUER with T2DM at baseline who achieved the following composite goals (CG) by Week 56: CG-A, WL [gt]5%, HbA1c [lt]6.5%, SBP [lt]130 mm Hg; and CG-B, WL [gt]10%, HbA1c [lt]6.5%, SBP [lt]130 mm Hg. Percentage of subjects with net changes (% increase minus % decrease) in concomitant antidiabetic and antihypertensive medications at Week 56 was also captured. Subjects were managed to T2DM standards of care (diet, exercise; metformin monotherapy was permitted at baseline; addition of oral hypoglycaemics by physicians blinded to assignment was permitted to achieve established HbA1c targets [ADA]). In total, 357 subjects with T2DM at baseline with [ge]1 postbaseline assessment of WL, HbA1c, and SBP were included: lifestyle intervention plus PBO (n=144), PHEN 7.5 mg/TPM ER 46 mg (7.5/46; n=63), or PHEN 15 mg/TPM ER 92 mg (15/92; n=150). At baseline, 66% were female, mean age was 52.6 years, mean weight was 100.9 kg, mean HbA1c was 6.8%, and 70.8% (n=253) were using metformin. Mean SBP was 125.9 mm Hg (40.3% of subjects had SBP [ge]130 mm Hg). At 56 weeks, CG-A was achieved by 11.8%, 27.0%, and 39.3% and CG-B by 4.2%, 14.3%, and 31.3% of the PBO, 7.5/46, and 15/92 groups, respectively ([italic]P[/italic][lt].0001 between treatment groups). For subjects achieving CG-A or CG-B, percentage of subjects with a net change in concomitant antidiabetic and antihypertensive medications was neutral or increased for PBO and decreased in PHEN/TPM ER[ndash]treated subjects ([italic]P[/italic]=NS between treatment groups). In these subjects with T2DM, there were 6 reports of hypoglycaemia: 4 (3 mild, 1 severe) in PBO, 1 (mild) in 7.5/46, and 1 (mild) in the 15/92 group; none led to study-drug discontinuation. Discontinuation due to adverse events (AEs) in this population was 8%, 9%, and 19% for PBO, 7.5/46, and 15/92, respectively, with the most common AEs being constipation and paraesthesia. These data indicate that the WL achieved by PHEN/TPM ER[ndash]treated subjects can drive improvements in HbA1c and SBP, significantly increasing the achievement of composite goals recommended for the management of T2DM.[br][br]Sources of Research Support: VIVUS, Inc.[br][br]Disclosures: DHR: Consultant, Vivus USA, Novo Nordisk, Takeda. WTG: Medical Advisory Board Member, Daiichi-Sankyo, Johnson & Johnson, Vivus USA, Abbott Laboratories; Clinical Researcher, Merck & Co., Amylin Pharmaceuticals, Vivus USA, Abbott Laboratories, Daiichi-Sankyo; Stockholder, Merck & Co., Genzyme Corporation, Vivus USA, Bristol-Myers Squibb, Amylin Pharmaceuticals; Speaker Bureau Member, Merck & Co., Abbott Laboratories. WWD: Employee, Vivus USA. 2012-06-23T16:30:00 362 2012-06-23T00:00:00 1899-12-30T16:30:00 535 101 899 S18-4 OR300-01 Saturday 58 2012


59 ENDO12L_S18-5 ORAL SESSION: Clinical Trial Symposium (3:45 PM-5:15 PM) Testosterone Dose-Response Relationships in Surgically Menopausal Women: Effects on Body Composition and Muscle Strength in a Randomized Trial Grace Huang, Shalender Bhasin, Thomas G Travison, Maithili Davda, Matthew Ho, Thomas W Storer, Renee Miciek, Phillip E Knapp, Lauren Collins, Monica Ursino, Connie Dzekov, Shehzad Basaria Boston University School of Medicine, Boston, MA; Charles R Drew University of Medicine and Science, Los Angeles, CA [bold]Background:[/bold] Menopause has been associated with a loss of muscle mass and an increase in fat mass. Testosterone therapy is being considered to improve body composition in menopausal women. However, the dose-response relationships between testosterone and the changes in body composition and muscle strength in women have not been established.[br][bold]Objective: [/bold]To determine the dose-dependent effects of graded doses of testosterone on body composition and muscle strength in surgically menopausal women.[br][bold]Methods: [/bold]71 surgically menopausal women received a standardized estrogen regimen during the 12-week run-in period, and were then randomized to one of 5 groups to receive weekly IM injections of placebo (n=15), 3 (n=14), 6.25 (n=14), 12.5 (n=15) or 25 mg (n=13) testosterone enanthate for 24 weeks. Total and free testosterone levels were measured by LC-MS/MS and equilibrium dialysis, respectively. Lean body mass (LBM) and fat mass were measured using DXA scan and muscle strength was assessed by one-repetition maximum method at baseline and week 24.[br][bold]Results: [/bold]71 women were randomized. At baseline, mean age was 53 yrs, BMI 30 kg/m[sup2], total testosterone 13.7 ng/dl and free testosterone 2.2 pg/ml. On-treatment nadir testosterone concentrations were 14, 87, 106, 164 and 211 ng/dl at the 0, 3, 6.25, 12.5 and 25-mg doses, respectively. Changes in LBM were highly correlated with changes in testosterone concentrations; the estimated between-person difference in LBM was 0.67 kg per 100 ng/dl change in testosterone (95% CI: 0.23, 1.11; p = 0.003). There was a significant increase in LBM in the 25-mg dose group (average increase=1.7 kg, p= 0.01). No significant changes in fat mass and leg press strength were seen.[br][bold]Conclusion: [/bold]Testosterone administration in surgically menopausal women was associated with dose and concentration-dependent gains in LBM. Long-term randomized trials are needed to determine whether clinically meaningful improvements in other outcomes can be achieved safely with testosterone doses that do not induce virilization or worsen cardiovascular risk.[br][br]Nothing to Disclose: GH, SB, TGT, MD, MH, TWS, RM, PEK, LC, MU, CD, SB 2012-06-23T16:45:00 362 2012-06-23T00:00:00 1899-12-30T16:45:00 1982 101 900 S18-5 OR300-01 Saturday 59 2012


60 ENDO12L_S18-6 ORAL SESSION: Clinical Trial Symposium (3:45 PM-5:15 PM) Carbohydrate Modified Diet and Insulin Sensitizers Reduce Fasting Insulin and Body Weight, Modulate Measures of the Metabolic Syndrome, and Improve Adipokines in EMPOWIR (Enhance the Metabolic Profile of Women with Insulin Resistance) a Randomized Clinical Trial (NCT00618071) of Normoglycemic, Hyperinsulinemic Women with Midlife Weight Gain Harriette Rosen Mogul, Ruth Freeman, Philipp E Scherer, Michael Frey, Feras M Hantash, Ron Greenbaum, Sheila Jozak, LeeAnn Klein, Karen Tanenbaum New York Medical College, Valhalla, NY; Einstein, Bronx, NY; University of Texas Southwestern Medical Center, Dallas, TX; Quest Diagnostics, San Juan Capistrano, CA; Westchester Medical Center, Valhalla, NY [bold]BACKGROUND [/bold]Progressive midlife weight gain is associated with diabetes[sup]1 [/sup]and all-cause mortality[sup]2[/sup] and preventive interventions targeting high risk populations are critically needed. [bold]EMPOWIR[/bold], a double blind, placebo controlled, randomized clinical trial of metformin (MF) 2000mg [amp] MF plus 4 mg (low dose) rosiglitazone (RSG) was conducted to evaluate prior research on Syndrome [bold]W,[/bold] an early manifestation of insulin resistance, defined by the triad of [bold]W[/bold]eight gain ([ge]20 lbs after the 20[apos]s), [bold]W[/bold]aist gain, and [bold]W[/bold]hite Coat hypertension and normoglycemic hyperinsulinemia[sup]3-6 [/sup]and test the [bold]hypothesis [/bold]that insulin sensitizers plus carbohydrate modified diet could reduce BW and attenuate risks of diabetes and related comorbidities. [bold]METHODS [/bold]We compared fasting insulin (FIn) [amp] glucose (FBS), BW, HOMA, blood pressure (BP), lipids, adiponectin (Adp), [amp] leptin (Lp) at baseline [amp] 6-months in 46 subjects (mean age 46.6, BMI 30.5kg/m[sup]2[/sup], A1C 5.4%) meeting inclusion criteria: age 35-55; 20 lb wt gain; normal GTT with [uarr]AUC-insulin. Paired t-tests [amp] multivariate models were used (SPSS 19.0). Participants attended 4 weekly nutrition workshops (lead-in phase) at 2 study sites to introduce the EMPOWIR dietary intervention: a food exchange program (40% carbohydrates-40% protein-20% fat) promoting increased intake of vegetables, low-glycemic index fruits, low-fat protein [amp] dairy products, elimination of free sugars, [amp] restriction of 3 allowable additional carbohydrates (starches) to after 4PM. [bold]RESULTS[/bold] Significant reductions in mean BW were observed in all 3 study arms ([italic]P[apos]s[/italic]=.049,.005,.017). FBS declined significantly only in the MF arm (89.4[rarr]84.0mg/dl, [italic]P[/italic]=.034) FIn reduction was significant in the MF arm (12.5[rarr]8.0[micro]U/ml, [italic]P[/italic]= .011). HOMA decreased in MF [amp] MF+RSG arms (2.5[rarr]1.6 [amp]1.9[rarr]1.3, [italic]P[apos]s=[/italic].054 [amp].013). SBP decreased in the MF arm (114.3[rarr]107.2, [italic]P[/italic]=.001). HDL increased in placebo [amp] MF arms (49.3[rarr]56.3 [amp] 57.4[rarr]67.7 mg/dl, [italic]P[apos]s[/italic]=.016 [amp].026) without other significant lipid changes. Adp increased only in the MF+RSG arm (11.2[rarr]18.5[micro]g/ml, [italic]P[/italic][lt].001). Lp did not decline in subjects with BW loss (mean difference -0.17, [italic]P[/italic]=.907). MF was well tolerated with no reported adverse events. [bold]CONCLUSIONS [/bold]The EMPOWIR diet independently, and in combination with insulin sensitizers, reduced BW in a diverse cohort of normoglycemic, hyperinsulinemic midlife women and merits additional study. Measures of glucose homeostasis and metabolic syndrome improved significantly with MF, whereas Adp increased only in the MF + RSG study arm.[br][br]1. Colditz GA, Willett WC, Rotnitzky A, Manson JE. Weight gain as a risk factor for clinical diabetes mellitus in women. Ann Intern Med 1995; 122(7):481-486. 2. Manson JE, Willett WC, Stampfer MJ et al. Body weight and mortality among women. N Engl J Med 1995; 333(11):677-685. 3. Mogul HR, Peterson SJ, Weinstein BI, Zhang S, Southren AL. Metformin and carbohydrate-modified diet: a novel obesity treatment protocol: preliminary findings from a case series of nondiabetic women with midlife weight gain and hyperinsulinemia. Heart Dis 2001; 3(5):285-292. 4. Mogul HR, Peterson SJ, Weinstein BI, Li J, Southren AL. Long-term (2-4 year) weight reduction with metformin plus carbohydrate-modified diet in euglycemic, hyperinsulinemic, midlife women (Syndrome W). Heart Dis 2003; 5(6):384-392. 5. Mogul HR, Weinstein BI, Mogul DB et al. Syndrome W: a new model of hyperinsulinemia, hypertension and midlife weight gain in healthy women with normal glucose tolerance. Heart Dis 2002; 4(2):78-85. 6. Mogul HR, Freeman R, Hantash FX. Long-term hormonal adaptations to weight loss. N Engl J Med 2012; 366.[br][br]Sources of Research Support: EMPOWIR (ClinicalTrials.gov:NCT00618071) is an unsolicited, investigator-initiated study supported by Glaxo Smith Kline, with additional support from a Clinical and Translational Science Award (CTSA) from the National Institutes of Health(UL11RR025740)awarded to the GCRC at Einstein of Yeshiva University.[br][br]Disclosures: HRM: Principal Investigator, GlaxoSmithKline. FMH: Study Investigator, Quest Diagnostics. Nothing to Disclose: RF, PES, MF, RG, SJ, LK, KT 2012-06-23T17:00:00 362 2012-06-23T00:00:00 1899-12-30T17:00:00 778 101 901 S18-6 OR300-01 Saturday 60 2012


61 ENDO12L_SAT-1 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Primary Amenorrhea Due to Endometrial Hypoplasia in Three Consanguineous Sisters Cicilia LR Santos, Larissa S Nazareno, Angelica A Amato, Adriana L Porto University of Bras[iacute]lia, Bras[iacute]lia, Brazil Introduction: primary amenorrhea (PA) comprises a broad range of possible clinical conditions. We describe herein 3 consanguineous sisters with PA associated with endometrial hypoplasia (EmH), a rare condition of unknown etiology. Case reports: three sisters were referred to the Endocrinology Unit because of primary amenorrhea. They had normal pubertal development, were born from consanguineous healthy parents, and had no past history of systemic or inflammatory pelvic disease. General physical and gynecological examination were unremarkable. Serum TSH and prolactin levels were all normal, and their karyotype was 46,XX. Pelvic ultrasonography showed a slightly diminished uterus ([lt]25 cm3). Serial hormones profile and pelvic ultrasonografic imaging (USG) over 4 weeks showed: Case 1:33 years-old(yo), sparse, irregular vaginal bleeding after past use of estrogen-progestagen oral contraceptive. Hormones profile: First weed(W1): FSH 10,07mIU/ml; LH 4,9mIU/ml; Estradiol(E) 7,0pg/mL; Progesteron(P) 0,24 ng/mL; USG: endometrium(Em) 1mm, ovarian follicles (OFs) [lt] 10mm. Second week(W2): FSH 7,54; LH 6,6; E 22,88; P [lt] 0,21; USG: Em 1mm, maximum follicular diameter(OFmax)= 12mm. Third week(W3): FSH 16,47; LH 37,6; E 46,82; P 0,75; USG: Em 3mm, corpus luteum (CL) of 13mm. Fourth week(W4): FSH 5,2; LH 4,3; E 54,61; P 8,77; USG: Em 3mm, OFs of 2 and 3mm. Case 2:30yo, denied past hormonal treatment. W1: FSH 9,56; LH 21,0; E 87,88; P [lt] 0,21; USG: thin Em and OFmax= 12mm. W2: FSH 8,57; LH 15,6; E 63,74; P 7,25; USG: Em 1mm, OFs [lt]5mm. W3: FSH 3,26; LH 3,9; E 138,18; P 5,69; USG: Em not detected, OFs [lt]6mm. W4: FSH 12,34; LH 10,20; E 53,74; P 0,58; USG: Em 2.2mm, OFs [lt]6mm. Case 3:31yo, denied past hormonal treatment, had a unilateral salpingectomy due to a tubal pregnancy. W1:FSH 7,92; LH 3,9; E 10,11; P 0,23; USG:thin Em and OFs [lt]1mm. W2: FSH 10,07; LH 8,0; E 7,0; P [lt]0,21; USG: Em 1mm and OFs[lt]7mm. W3: FSH 7,89; LH 7,9; E 35,78; P [lt]0,21; USG:Em 1mm and OFmax =10mm. W4: FSH 6,13; LH 14,9; E 436,32; P 0,34; TV USG:Em 2.4mm, image compatible with cystic CL. Conclusions: We observed a hormonal cyclic pattern with P levels on the ovulatory range and USG image of CL in 2 of the 3 cases, whereas thin Em appearance remained unchanged. Altogether, these findings suggest EmH, probably associated with a monogenic disturbance. Possible candidates would be PAX8, a transcriptional factor involved in mullerian duct development, and WNT4, which has been associated with Rokitansky syndrome.[br][br]Nothing to Disclose: CLRS, LSN, AAA, ALP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1544 49 122 SAT-1 PO32-01 Saturday 61 2012


62 ENDO12L_SAT-2 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Fetal Sulfate Supply in Human Gestation Paul A Dawson, Scott Petersen, Robyn L Rodwell, Phillip Johnson, Joanna Rakoczy, Francesca Grace, David G Simmons, H David McIntyre Mater Medical Research Institute, South Brisbane, Australia; Mater Mothers Hospital, South Brisbane, Australia; Mater Health Services, South Brisbane, Australia; University of Queensland, St Lucia, Australia; University of Queensland, South Brisbane, Australia Sulfate is an important nutrient for numerous cellular and metabolic processes in fetal development, including biotransformation of iodothyronines and steroids. In human gestation, maternal serum sulfate levels increase by approximately 2-fold, which enhances sulfate availability to the growing fetus. Sulfate is supplied from maternal circulation to the fetus via placental sulfate transporters. In this study, we measured venous cord plasma sulfate levels from healthy term pregnancies (altruistic cord blood donors), as well as maternal plasma sulfate levels in early ([lt]20wks) and late ([gt]30wks) gestation. Maternal plasma sulfate levels increased from (394[plusmn]12uM, n=43) in early pregnancy to (429[plusmn]12uM, n=33) in late gestation. Cord blood sulfate levels were lower in male babies (325[plusmn]16uM, n=31) when compared to female babies (371[plusmn]13uM, n=42). To build a model of directional sulfate transport in the placenta, we determined the relative abundance and spatial expression of all 10 sulfate transporter mRNAs in placentae (n= 10 male and 6 female babies) from uncomplicated healthy pregnancies at elective caesarean section. Quantitative real time PCR was used to determine the relative abundance of sulfate transporter mRNAs, and the spatial expression of mRNAs was determined by [italic]in situ[/italic] hybridisation with gene-specific DIG-labelled riboprobes. We detected very strong mRNA expression of [italic]SLC13A4[/italic] and [italic]SLC26A2[/italic], and intermediate levels of [italic]SLC26A1[/italic], [italic]SLC26A6[/italic] and [italic]SLC26A11[/italic]. Placental [italic]SLC13A4[/italic] mRNA was localised specifically to syncytiotrophoblasts, whereas [italic]SLC26A2[/italic] was detected in the cytotrophoblast cells. In summary, these data show high maternal plasma sulfate levels in late gestation when high sulfate levels are required for fetal development. In addition we show lower cord plasma sulfate levels from male babies when compared to female babies at birth, suggesting gender differences in sulfate requirements or placental sulfate transport. We identified [italic]SLC13A4[/italic] and [italic]SLC26A2[/italic] as the most abundant sulfate transporter mRNAs expressed in human placentae, suggesting a role for these 2 transporters in supplying sulfate from mother to fetus.[br][br]Sources of Research Support: Mater Medical Research Institute. Mater Mothers[apos] Hospital Golden Casket Research Grant.[br][br]Nothing to Disclose: PAD, SP, RLR, PJ, JR, FG, DGS, HDM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 86 49 123 SAT-2 PO32-01 Saturday 62 2012


63 ENDO12L_SAT-3 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Prolonged Pregnancy in Women Is Associated with Attenuated Myometrial Expression of Progesterone Receptor Co-Regulator Kr[uuml]ppel-Like Factor 9 John Mark P Pabona, Daying Zhang, David S Ginsburg, Frank A Simmen, Rosalia CM Simmen University of Arkansas for Medical Sciences, Little Rock, AR; Crozer Chester Medical Center, Upland, PA; Arkansas Children[apos]s Hospital Research Institute, Little Rock, AR Post-term pregnancy represents 3-5% of all complications during delivery and leads to both maternal and neonatal morbidity and mortality. The signaling mechanism(s) that maintains uterine quiescence during pregnancy and initiates the timely induction of uterine contractility at term remains unclear. Kr[uuml]ppel-like factor 9 (KLF9), an Sp-family member, is a progesterone receptor (PGR) interacting protein in uterine cells. In mice, KLF9 is robustly expressed in myometrial circular and longitudinal layers during late pregnancy. Further, mice null for [italic]Klf9[/italic] exhibit delayed parturition with increased gestational length and incidence of neonatal deaths. To examine the contribution of KLF9 in human parturition, we evaluated myometrial biopsies obtained from women during delivery, following procedures approved by the Institutional Board of Crozer Chester Medical Center and with signed consent. Participants with gestational age of 37 to [lt] 41 weeks were considered to have term pregnancy, while those with [ge]41 weeks were considered to exhibit prolonged pregnancy. Gene expression analysis showed that myometria from women with prolonged pregnancy (n=5) had decreased levels of KLF9, PGR-B and myosin light chain kinase transcripts, when compared to those of women with term pregnancy (n=8). The reduction in transcript levels was confirmed at the level of nuclear KLF9, PGR-A and PGR-B proteins and was accompanied by lower PGR-A/PGR-B ratio (P[lt]0.05). The loss of total PGR expression in myometria of patients with prolonged, relative to those with term pregnancies was also confirmed by immunohistochemistry. Estrogen receptor (ER) [alpha] expression, both at the level of mRNA and nuclear protein, was also lower for myometria of women with prolonged vs. term pregnancies. Myometrial levels of total and phosphorylated NF-[kappa]B (p65) proteins as well as of PGF2[alpha] receptor transcripts were similarly attenuated in women with prolonged relative to those with term pregnancies. In contrast, transcript levels for oxytocin receptor and inducible nitric oxide synthase did not differ between the two groups. Loss of myometrial KLF9 may aberrantly influence PGR, ER[alpha] and NF- [kappa]B signaling networks to promote uterine quiescence and delay labor. Our findings suggest that prolonged pregnancy associated with a deficiency in KLF9 expression may be a conserved phenomenon in mice and humans and importantly, that KLF9 has a physiological and pathological role in the timely onset of parturition.[br][br]Sources of Research Support: NIH-HD21961.[br][br]Nothing to Disclose: JMPP, DZ, DSG, FAS, RCMS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 240 49 124 SAT-3 PO32-01 Saturday 63 2012


64 ENDO12L_SAT-4 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Incident Metabolic Syndrome Is Not Increased in Women with Hyperandrogenemic Oligomenorrhea across Menopausal Transition: Longitudinal Follow-Up from the Study of Women[apos]s Health Across the Nation (SWAN) Alex J Polotsky, Amanda Allshouse, Sybil L Crawford, Sioban D Harlow, Naila Khalil, Rasa Kazlauskaite, Nanette Santoro, Richard Legro University of Colorado, Aurora, CO; University of Massachusetts, Worcester, MA; University of Michigan, Ann Arbor, MI; Boonshoft School of Medicine, Wright State University, Dayton, OH; Rush University Medical Center, Chicago, IL; Pennsylvania State College of Medicine, Hershey, PA Reproductive-age women with oligomenorrhea (Oligo) and hyperandrogenemia (HA), features of Polycystic Ovary Syndrome (PCOS), exhibit adverse cardiovascular risks as compared to age-matched controls. Recent data suggest that menstrual irregularity(1), HA(2) and insulin resistance(3) may improve with aging in PCOS women approaching menopause. Cross-sectional analysis of 2543 SWAN women at baseline showed that HA but not history of Oligo was independently associated with the risk of prevalent metabolic syndrome (MetS) (4). Persistence of metabolic risks after menopause is not well understood.[br][bold]Objective[/bold]: To evaluate metabolic health in women with history of HA and Oligo across menopausal transition.[br][bold]Methods[/bold]: SWAN is a multi-ethnic cohort of over 3000 US women as they traverse menopause with mean age at baseline of 45.8 years. Current analysis includes 8-10 years of follow-up. History of [ge] 2 instances of non-gestational/non-lactational amenorrhea for [ge] 3 months was classified as Oligo. The highest tertile of serum testosterone was set as HA. MetS was defined by NCEP ATP III criteria. Cox Hazard Ratios (HR) of MetS were estimated, with adjustment for age, ethnicity, BMI, smoking, menopausal stage [amp] study site.[br][bold]Results[/bold]: Among 1932 SWAN subjects who were MetS-free at baseline, 409 new cases of MetS were identified during 12,363 woman-years of follow-up over 8 years. Mean age at natural menopause was 51.3 years among 734 women who were at least 12 months past their final menstrual period. Women with HA and Oligo developed incident cases of MetS at a similar rate compared to their counterparts: HR of 1.1 (0.9-2.3) vs. eumenorrhea and normal androgens; HR of 1.2 (0.7-2.0) vs. Oligo and normal androgens; HR of 1.2 (0.7-1.9) vs. HA and eumenorrhea. Women with HA and Oligo had a similar trajectory of change for all individual components comprising MetS. Smoking and obesity were the strongest predictors of incident MetS: HR of 1.5 (1.1-1.9) for current smoking [amp] HR of 1.8 (1.7-1.9) per each 5 units of BMI.[br][bold]Conclusions[/bold]: While women acquire MetS at a more rapid rate after menopause, SWAN participants with history of HA and Oligo did not develop incident MetS more frequently than those without these characteristics. This suggests that history of androgen excess and menstrual irregularity in early life does not remain an independent risk for metabolic health after menopause. Attention to modifiable risks of obesity and smoking is warranted regardless of the PCOS diagnosis.[br][br](1)Elting MW et al., Human Reproduction 2000; 15:24. (2)Winters SJ et al., Fertil Steril 2000. 73: 724. (3)Brown ZA et al., Fertil Steril 2011; 96: 1259. (4)Polotsky AJ et al., Fertil Steril 2011; 96: S42.[br][br]Sources of Research Support: SWAN has grant support from the NIH, DHHS, through the NIA, the NINR and the ORWH (Grants NR004061; AG012505, AG012535, AG012531, AG012539, AG012546, AG012553, AG012554, AG012495). The content of this abstract is solely the responsibility of the authors and does not necessarily represent the official views of the NIA, NINR, ORWH or the NIH.[br][br]Nothing to Disclose: AJP, AA, SLC, SDH, NK, RK, NS, RL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 276 49 125 SAT-4 PO32-01 Saturday 64 2012


65 ENDO12L_SAT-5 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Aromatase Inhibition Normalizes Luteal Function but Dramatically Reduces Estrone Conjugate Excretion in Obese Women Lauren A Ross, Alex J Polotsky, Alexander Kucherov, Andrew P Bradford, Jennifer Lesh, Justin Chosich, Nanette Santoro University of Colorado Denver, Aurora, CO; Albert Einstein College of Medicine, Bronx, NY Adult female obesity is associated with menstrual cycle irregularities, subfertility and a hypogonadotropic phenotype. We have previously shown that aromatase inhibitor (AI)-treated obese women exhibit increased gonadotropin production similar to untreated normal weight women (1).[br][bold]Objective:[/bold] To assess the mechanism of action of AI in obesity, we examined reproductive hormone excretion across the menstrual cycle in obese women compared to normal weight women, after treatment with an AI.[br][bold]Methods:[/bold] We administered a 7-day, weight-adjusted dose of letrozole to 22 regularly cycling women: 12 obese (BMI, 37.1[plusmn]7.0kg/m2) and 10 normal weight (BMI, 21.2[plusmn]1.3kg/m2). Urine was collected daily over the course of the treatment cycle and assayed for luteinizing hormone (LH) [amp] follicle stimulating hormone (FSH; using the DELFIA immunofluorometric assay platform), and estrone conjugates (E1c) [amp] pregnanediol glucuronide (Pdg; using an in-house ELISA). All hormones were normalized to creatinine, and the day of ovulation was determined using a validated algorithm (2).[br]LH, FSH, E1c and Pdg were estimated for the whole cycle as well as follicular and luteal phases by computing sums of individual hormones. Groups were compared by t and Mann-Whitney tests as appropriate.[br][bold]Results[/bold]: Age did not differ between obese and normal weight women (30.5[plusmn]4.3 years vs. 30.9[plusmn]1.0 years).[br]Whole cycle E1c was half the level of controls in the obese women [331(259-465) mIU/mg Cr vs. 775(695-2180) mIU/mg Cr for obese and normal weight groups, respectively, p[lt]0.01] as was average E1c (p[lt]0.01) and peak E1c (p[lt]0.01).[br]Whole cycle Pdg (12.3 (5.2-16.2) ug/mgCr vs. 12.5 (9.2-33.8) ug/mgCr, p=0.47); LH (242 [plusmn]144 mIU/mg Cr vs. 335[plusmn]166 mIU/mg Cr, p=0.18); FSH (179[plusmn]95 mIU/mg Cr vs. 137[plusmn]59 mIU/mg Cr, p=0.23) did not differ between obese and normal weight women, respectively, nor did they differ by follicular or luteal phase means for these hormones.[br][bold]Conclusions[/bold]: Normalization of gonadotropin output and luteal function occurs at the expense of drastically reduced E1c excretion in AI-treated obese women as compared to normal weight AI-treated controls. These findings suggest that the adiposity-related reproductive phenotype is receptive to modulation of estrogen dynamics. Obese women demonstrate increased sensitivity to interruption of estrogen feedback with AI, implying that female obesity is a state of enhanced estrogen negative feedback at the hypothalamic-pituitary level.[br][br]1. Polotsky AJ et al. Endocr Rev 2011;32:S12. 2. Santoro N et al. JCEM 1996;81:1495.[br][br]Sources of Research Support: NIH U54 HD058155 Center for the Study of Reproductive Biology; K24 HD041978 to NS; 1UL1 RR025780 (University of Colorado CTRC).[br][br]Nothing to Disclose: LAR, AJP, AK, APB, JL, JC, NS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 902 49 126 SAT-5 PO32-01 Saturday 65 2012


66 ENDO12L_SAT-6 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Effects of Vaginal Estriol Cream on Circulating Hormones in Normal Cycling Women K-Y Francis Pau, Heidi Jones, Ilana Dzuba, Susan A Ballagh, Tracy Irwin, Bryna Harwood, Wesley H Clark, Yael Swica, Beverly Winikoff Oregon National Primate Research Center, Oregon Health [amp] Sciences University, Beaverton, OR; CUNY School of Public Health, Hunter College, New York, NY; Gynuity Health Projects, New York, NY; Los Angeles County Harbor-UCLA, Torrance, CA; University of Illinois at Chicago, Chicago, IL; New York Presbyterian Hospital, New York, NY; Columbia University College of Physicians and Surgeons, New York, NY [bold]Background:[/bold] Studies in ovariectomized non-human primates suggest that estriol (E3) administered topically protects against SIV transmission (1). In humans, HIV transmission is higher in premenopausal women during the luteal phase and pregnancy when progesterone (P4) levels are high (2). As part of a Phase 1 randomized, placebo-controlled, double-blinded trial to evaluate the effect of a topical E3 cream on vaginal health, this report examines the cream[apos]s effect on systemic hormones.[br][bold]Methods:[/bold] Of the 102 eligible consenting women aged 18-40 years, 82 completed 4 study visits in both follicular and luteal phases before and after randomization to 4.0mg estriol cream (1mg estriol/1ml cream) (n=38) or matching placebo (n=44). A single blood sample was taken at each visit for detection of estrone (E1), estradiol (E2), E3, P4, luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Data were analyzed with SPSS and compared by paired rank sum test.[br][bold]Results:[/bold] Serum E3 was non-detectable ([lt]0.07 ng/ml) in all subjects during baseline and among those assigned to placebo. E3 cream increased serum E3 level in 10 subjects (26.3%) during the follicular phase (0.09 +/- 0.01 ng/ml, range 0.08-0.23 ng/ml) and 11 (28.92%) during the luteal phase (0.09 +/- 0.01, range 0.08-0.30 ng/ml). None of the other 5 hormonal levels changed (p[gt]0.05) from baseline in the follicular phase. During the luteal phase, P4 levels were lower with E3 cream compared to placebo (median: 1.24 vs 8.69 ng/ml, p[lt]0.009; mean: 3.92 vs 7.17 ng/ml, p[lt]0.004). Of the 11 subjects with increased E3, 6 had P4 levels below the mean. Baseline P4 levels during the luteal phase did not differ between the two groups (median: 5.64 vs 6.60 ng/ml, p= 0.61).[br][bold]Conclusions:[/bold] Consistent with findings in non-human primates, these results show that a small amount of E3 can be found in the circulation of less than 30% of users in either the follicular or luteal phase. While vaginal E3 cream has little effect on circulating E1, E2, LH and FSH, serum P4 levels are reduced during the luteal but not the follicular phase. The relationship of E3 and P4 levels and their association with menstrual cycle changes remain to be explored.[br][br](1)Smith SM, Mefford M, Sodora D, Klase Z, Singh M, Alexander N, Hess D, Marx PA. Topical estrogen protects against SIV vaginal transmission without evidence of systemic effect. AIDS. 2004 Aug 20;18(12):1637-43. (2)Mishell DR, Davajan V. Infertility, contraception, and reproductive endocrinology. Montvale, NJ: Medical Economics, 1986.[br][br]Sources of Research Support: The United States Agency for International Development (USAID) through the International Partnership for Microbicides.[br][br]Nothing to Disclose: K-YFP, HJ, ID, SAB, TI, BH, WHC, YS, BW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 931 49 127 SAT-6 PO32-01 Saturday 66 2012


67 ENDO12L_SAT-7 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Factors within the Intravascular Compartment Associate with Progression of Brain White Matter Hyperintensity in Women of the Kronos Early Estrogen Prevention Study (KEEPS) Limor Raz, Kejal Kantarci, Nirubol Tosakulwong, Timothy G Lesnick, Muthuvel Jayachandran, Samantha M Wille, Matthew C Murphy, Matthew L Senjem, Clifford R Jack, Virginia M Miller Mayo Clinic, Rochester, MN; Mayo Clinic, Rochester, MN; Mayo Clinic, Rochester, MN; Mayo Clinic, Rochester, MN White matter hyperintensities (WMH) on brain MRI correlate with a future risk for mild cognitive impairment in elderly populations of men and women. Relationships of WMH with vascular disease in younger but vulnerable populations such as menopausal women remain unexplored. Our objective was to investigate relationships among conventional and novel cardiovascular (CV) risk factors regarding the presence and progression of WMH in recently menopausal women (within 6 months [ndash] 3 yrs of their last menses) enrolled in the Kronos Early Estrogen Prevention Study (KEEPS) at Mayo Clinic (n=95). Conventional CV risk factors (systolic blood pressure, plasma lipids, glucose, triglycerides and body mass index) and activated cell membrane-derived microvesicles (MVs) were assessed in each woman prior to brain MRI at baseline (prior to randomization to treatment groups), and at 36 and 48 months (mo) after randomization. MRI images included a Fluid Attenuated Inversion Recovery (FLAIR) sequence from which WMH volumes were calculated using a semi-automated segmentation algorithm. Changes in WMH volume were quantified from baseline to 36 and 48 mo. Linear regression models adjusted for age, time past menopause and APOE [epsilon]4 status were used to test for association of CV factors with baseline WMH and changes in WMH. The median [inter-quartile range (IQR)] of WMH at baseline was 1909 (1538, 2655) mm[sup]3[/sup]. Volumes of WMH were not associated with conventional CV risk factors at baseline. Volumes of WMH tended to increase over time; median (IQR) volume changes were 101 (-165, 510) mm[sup]3[/sup] and 155 (-89, 567) mm[sup]3[/sup] at 36 and 48 mo, respectively. These increases in WMH associated with numbers of platelet-derived and thrombogenic MVs, (those positive for phosphatidylserine) at baseline, but not with conventional CV risk factors. These results suggest that activated platelets and cell membrane-derived thrombogenic MVs within the vascular compartment may affect development of WMH within the brain as has been suggested for their involvement in anatomical changes in other organs such as arterial intimal medial thickening and arterial calcification in the heart.[br][br]Sources of Research Support: The Aurora Fnd, NIH HL90639, NS066147, HD65987 and the Mayo Fnd.[br][br]Nothing to Disclose: LR, KK, NT, TGL, MJ, SMW, MCM, MLS, CRJ, VMM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 568 49 128 SAT-7 PO32-01 Saturday 67 2012


68 ENDO12L_SAT-8 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) The Pathogenic Role of Novel Steroid Receptor Coactivator-1 C-Terminal Isoform in Endometriosis Progression Sang Jun Han, Shannon M Hawkins, Khurshida Begum, Sung Yun Jung, Ertug Kovanci, Jun Qin, John P Lydon, Francesco J DeMayo, Bert W O[apos]Malley Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX Approximately 10% of women of reproductive age in the United States suffer from the severe symptoms of endometriosis. However, almost nothing is known of its molecular etiology. Although endometriosis is considered as an estrogen-dependent inflammatory disease, the role of coactivators (SRCs), key regulators of estrogen receptor signaling, has not been elucidated in endometriosis progression. Here, we reveal that a previously undiscovered isoform of SRC-1 works as a [apos]causative molecular regulator[apos] in endometriosis progression and has the following unique molecular properties:[bold]1) [/bold]A novel 70-kDa SRC-1 proteolytic isoform has been discovered that migrates to the cytoplasm and is highly elevated in endometriotic tissue of mice with surgical induced endometriosis - and also in human endometriotic stromal cells isolated from ectopic lesions of women with endometriosis; [bold]2)[/bold] TNFalpha activates the MMP9 protease to process SRC-1 to this 70-kDa C-terminal isoform (791-1441 aa.) within endometriotic tissue; [bold]3)[/bold] To promote endometriotic tissue growth, the processed 70-kDa SRC-1 C-terminal isoform then acts mechanistically by protecting human endometrial cells from TNFalpha-induced [apos]apoptosis[apos] by directly interacting with, and inhibiting, procaspase 8 and importantly, it also enhances the epithelial-mesenchymal transition of human endometrial cells to promote their invasive capacity. Thus, this TNFalpha/MMP9/SRC-1-isoform axis functions as a newly discovered pathogenic pathway for endometriosis progression; the isoform could be employed as a molecular marker and/or therapeutic target for endometriosis.[br][br]Sources of Research Support: R01 HD08188 to B. W. O[apos]Malley and U54HD007495 pilot grant to S.J. Han.[br][br]Nothing to Disclose: SJH, SMH, KB, SYJ, EK, JQ, JPL, FJD, BWO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 588 49 129 SAT-8 PO32-01 Saturday 68 2012


69 ENDO12L_SAT-9 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Racial/Ethnic Differences in Sex Hormone Levels among Postmenopausal Women in the Diabetes Prevention Program (DPP) Catherine Kim, Sherita Golden, Kieren Mather, Gail A Laughlin, Shengchun Kong, Bin Nan, Elizabeth Barrett-Connor, John Randolph, Diabetes Prevention Program (DPP) University of Michigan, Ann Arbor, MI; Johns Hopkins University, Baltimore, MD; Indiana University, Indianapolis, IN; University of California, San Diego, La Jolla, CA; George Washington University, Washington, DC Context: Studies in mid-life women conflict regarding racial/ethnic differences in sex hormones: sex hormone binding globulin (SHBG), total and bioavailable estradiol (E2), total and bioavailable testosterone (T), and dehydroepiandrosterone (DHEA). Such differences may contribute to racial/ethnic variation in the incidence and consequences of sex hormone-sensitive comorbidities in older women, such as gynecological malignancies.[br]Objectives: We compared sex hormones by race/ethnicity in postmenopausal non-Hispanic white (NHW), Hispanic, and African-American (AA) women in the Diabetes Prevention Program (DPP).[br]Methods: The DPP was a randomized controlled trial of intensive lifestyle modification vs. metformin (850 mg twice a day) vs. placebo for diabetes prevention. We examined postmenopausal glucose-intolerant women participating in the DPP who did (n=310) or did not (n=370) use oral estrogen, with and without progestin, at baseline and 1-year follow-up. Postmenopause was defined as the absence of menses at baseline for at least one year without gynecologic surgery, bilateral oophorectomy, or age [gt]= 55 years and hysterectomy. Main outcome measures were baseline and 1-year change in serum levels of SHBG, total and bioavailable E2 (bioE2), total and bioavailable T, and DHEA.[br]Results: Among women not using estrogen, NHW had higher baseline total and bioavailable E2 and T levels than Hispanic women independent of age, type of menopause, waist circumference, alcohol intake, and current smoking. NHW also had higher levels of baseline bioE2 and lower levels of SHBG than AA. Among oral estrogen users, NHW had higher baseline total E2 and bioE2 than Hispanics, and higher levels of SHBG than AA independent of the same covariates. At 1-year, after adjustment for randomization arm, change in waist circumference, and the above covariates, NHW women not using estrogen had larger declines in total E2 and bioE2 levels than AA. Changes in sex hormones at 1-year did not differ by race/ethnicity among women using estrogen.[br]Conclusions: Among postmenopausal glucose-intolerant women, there were significant race/ethnicity differences in sex hormones and changes in sex hormones. Higher sex hormone levels in NHW women are consistent with reports of a higher incidence of hormone-sensitive cancers in NHW women compared to Hispanics and AA. Whether the greater reduction in E2 at 1-year in NHW results in a preferential reduction in risk in this group warrants further study.[br][br]Sources of Research Support: NIH (NIDDK).[br][br]Nothing to Disclose: CK, SG, KM, GAL, SK, BN, EB-C, JR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 614 49 130 SAT-9 PO32-01 Saturday 69 2012


70 ENDO12L_SAT-10 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Relations between Soluble P-Selectin and Sex Hormones in Postmenopausal Women in the Framingham Offspring Study Pornpoj Pramyothin, Thomas G Travison, Shalender Bhasin, Andrea D Coviello Boston University School of Medicine and Boston Medical Center, Boston, MA; Boston University School of Medicine and Boston Medical Center, Boston, MA Context: Soluble P-selectin (sP-sel), a cell-adhesion molecule with key roles in inflammation and coagulation, may contribute to cardiovascular disease (CVD) risk. sP-sel levels are higher in postmenopausal women but lower in women taking hormone therapy (HT). The relations between sP-sel and sex hormone levels in post-menopausal women is not understood.[br]Objective: To determine the relations between circulating levels of sP-Sel and endogenous sex hormones in postmenopausal women not taking HT: total (TT) and free (FT) testosterone, total (E2) and free (FE2) estradiol, total (E1) and free (FE1) estrone and their primary binding protein sex-hormone binding globulin (SHBG).[br]Methods: Multivariable linear regression models were used to determine the cross-sectional associations between circulating sP-sel and endogenous sex hormones in postmenopausal women who attended exam 7 (1998-2001) of the Framingham Offspring Study (FOS) who were not taking HT (n=875). Models were adjusted for factors associated with sex hormones and CVD risk including age, body mass index (BMI), smoking, insulin resistance, hypertension, cholesterol levels and statin use. Due to the inflammation associated with diabetes (DM), subgroup analyses were performed in women without DM (n=763). sP-sel was measured by R[amp]D enzyme-linked immunoassay; SHBG by immunoassay(Delfia-Wallac); TT, E2, and E1 by LC-MSMS; and FT, FE2, and FE1 were calculated.[br]Results: Mean[plusmn]SD age was 65[plusmn]8 yrs and mean[plusmn]SD BMI 28.0[plusmn]5.7 kg/m2. The prevalence of hypertension was 38%, hyperlipidemia 22%, metabolic syndrome 40%, diabetes 12.8%, and CVD 4%. In adjusted analyses, a 10% increase in sP-sel was associated with a 0.9% decrease in SHBG (p=0.03) and a 1.8% increase in E2 (p[lt]0.01) but not with changes in E1 (p=0.1). Further adjustment for SHBG did not alter results. A 10% increase in sP-sel was associated with a 2.4% increase in FE2 (p[lt]0.01) and 1.2% increase in FE1 (p=0.02). Adjustment for SHBG attenuated these associations to 1.9% (p[lt]0.01) for FE2 and 1% (p=0.04) for FE1. sP-sel was not associated with TT or FT in adjusted models. Exclusion of women with DM yielded similar results.[br]Conclusions: Circulating sP-sel levels were negatively associated with SHBG and positively associated with total and free E2 as well as FE1 in postmenopausal women not on HT. Defining the connection between inflammation and coagulation pathways and estrogens will lead to a better understanding of CVD in post-menopausal women.[br][br]Sources of Research Support: NIH Grant RO1 HL094755 (to A.D.C., S.B., and R.S.V.).[br][br]Nothing to Disclose: PP, TGT, SB, ADC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 660 49 131 SAT-10 PO32-01 Saturday 70 2012


71 ENDO12L_SAT-11 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Anti-Mullerian Hormone and Its Specific Receptor Are Differentially Regulated by Gonadotropins in Normo and Dysovulatory Women with the Polycystic Ovarian Syndrome Alice Pierre, Maeliss Peigne, Michael Grynberg, Nassim Arouche, Joelle Taieb, Laetitia Hesters, Jean-Yves Picard, Didier Dewailly, Renato Fanchin, Sophie Catteau-Jonard, Nathalie di Clemente INSERM, Clamart, France, Metropolitan; Universit[eacute] de Lille II, Lille, France, Metropolitan Anti-Mullerian hormone (AMH) is a member of the transforming growth factor family well known for its role during male sexual differentiation. However, AMH and its specific receptor AMHR-II are also expressed by granulosa cells of growing follicles. In rodents, AMH exerts a repressive role on folliculogenesis, but up to now, this effect has not been confirmed in human. These last years, serum AMH has been recognized as a useful tool in reproductive endocrinology, as a reliable marker of the ovarian follicular status and predictor of the ovarian response to controlled ovarian hyperstimulation. In particular, serum AMH is elevated in women with the polycystic ovarian syndrome (PCOS), the most common cause of female infertility, which is characterized by an increased number of small follicles. We had previously shown that AMH and AMHR-II expression are up-regulated in granulosa cells from PCOS women undergoing in vitro fertilization treatment, in keeping with a repressive role of AMH on follicle growth in these women. In this work, using real-time RT-PCR and ELISA experiments, we compared the regulation by gonadotropins of AMH and AMHR-II expression in primary culture of granulosa cells from control and normo and dysovulatory PCOS women. We show that AMH mRNAs are up-regulated by FSH in the three groups of women (+70% p=0.0068 and p=0.0469 for control and dysovulatory PCOS women respectively, and +40% p=0.0313 for normo ovulatory PCOS women), whereas there are stimulated by LH only in granulosa cells from dysovulatory PCOS women (+120% p=0.0391). Similarly, AMHR-II expression is unaffected by FSH in the three groups of women, whereas it is down-regulated by LH in control and normo ovulatory PCOS women (-30% p=0.0049 and -40% p=0.0469 respectively), but not in dysovulatory ones. Because LH levels are often elevated in PCOS women, our results could explain the increase of AMH expression by their granulosa cells. The reason why LH does not repress AMHR-II expression in dysovulatory PCOS cells is unknown. Altogether, our results demonstrate that LH regulation of the AMH/AMHR-II system is altered in PCOS granulosa cells, maintaining them in an immature stage. Because this dysregulation is associated with dysovulation in PCOS women, our results are in keeping with a role of AMH in this process.[br][br]Nothing to Disclose: AP, MP, MG, NA, JT, LH, J-YP, DD, RF, SC-J, NdC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 926 49 132 SAT-11 PO32-01 Saturday 71 2012


72 ENDO12L_SAT-12 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Impact of [italic]MTHFR[/italic] Polymorphisms C677T and A1298C in the Synthesis of Homocysteine and Follicular Estradiol Aline Amaro Santos, Bianca Bianco, Priscila Queiroz, Juliana Christofolini, Rene Busso, Denise Maria Christofolini, Caio Parente Barbosa Faculdade de Medicina do ABC, Santo Andre, Brazil [bold]Introduction: [/bold][italic]MTHFR[/italic] gene is essential in the folate metabolism, a major factor in the methylation reactions. Hypo or hypermethylation have been associated with dietary deficiencies of folate and genetic changes as polymorphisms of [italic]MTHFR[/italic] gene. The most common polymorphisms for this gene are C677T and A1298C and can also be linked to changes in the concentrations of estrogens. It was suggested that patients undergoing assisted reproduction techniques that carry the polymorphism had significantly lower concentrations of serum estrogen and follicle producing significantly fewer oocytes retrieved since the hormonal and biochemical content has great influence on the development and quality oocyte. We aimed to correlate levels of estradiol in blood and follicular fluid of infertile women with endometriosis and unexplained Infertility and polymorphism in [italic]MTHFR [/italic]gene, and to correlate the data with indicators of embryo quality. [bold]Material[/bold] [bold]and methods[/bold]: Eleven women that had unexplained infertility and forty patients that present endometriosis were studied. These patients underwent assisted reproduction treatment. The analysis of blood and follicular fluid was performed by measuring the concentrations of homocist[iacute]ne and estradiol and by the evaluation of molecular genetic polymorphisms C677T and A1298C [italic]MTHFR[/italic] gene from peripheral blood DNA. We used a control group composed by couples with male cause of infertility for comparison of molecular genetic analysis and serum and follicular homociste[iacute]ne and estradiol concentration. [bold]Results[/bold][bold]:[/bold] The groups were defined through the molecular genetic testing and the results were related to the findings serum and follicle estradiol. The distribution of genotypes revealed no differences between groups of infertile patients and controls (p=0.930 for serum and p=0.237 for follicle) for the C677T polymorphism and (p=0.603 for serum and p=0.550 for follicle) for polymorphism A1298C. Also, no differences were found concerning the concentration of homoc[iacute]steine between the three different genotypes. Embryo quality and embryo development showed no difference. [bold]Conclusion:[/bold] In this study was not observed a significant difference in the synthesis of serum estradiol and follicle in the presence of polymorphisms for C677T and A1298C of [italic]MTHFR[/italic] gene. There is no negative impact of these polymorphisms in the parameters of oocyte quality and embryonic development suggesting that the polymorphisms studied are not correlated with lower rates of pregnancy.[br][br]Sources of Research Support: FAPESP No. 2011/16274-4.[br][br]Nothing to Disclose: AAS, BB, PQ, JC, RB, DMC, CPB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1040 49 133 SAT-12 PO32-01 Saturday 72 2012


73 ENDO12L_SAT-13 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Pregnancy Loss and Neonatal Outcomes in Women with Thyroid Dysfunction in the First Trimester of Pregnancy Kewal Aditya, Varun Gupta, Shweta Achint, Tapasya Dhar, Jubbin Jagan Jacob, Kumkum Awasthi Christian Medical College and Hospital, Ludhiana, India; Christian Medical College and Hospital, Ludhiana, India Context: Universal screening of thyroid functions using both TSH assays and FT4 estimation in pregnancy leads to at least five different common diagnostic entities. Pregnancy and foetal outcomes in overt hypothyroidism and thyrotoxicosis is well documented, while outcomes with subclinical hypothyroidism and isolated hypothyroxemia are less clear.[br]Objective: To evaluate the pregnancy and neonatal outcomes in women with first trimester maternal thyroid dysfunction.[br]Design: The study is part of a prospective study evaluating routine screening of thyroid function tests among 1000 pregnant women in the first trimester (Thyroid Screening in Urban Ludhiana in Pregnancy Study: TULIPS). Normal first trimester values for TSH were considered to be between 0.03 to 2.3 mIU/L based on data from Chinese patients using the same automated chemiluminescent assay.(1) Normal non-pregnant range for free T4 was used for classification. The patients were classified based on the above ranges and compared to the reference group (antibody negative euthyroid patients).[br]Setting: The study was conducted in the antenatal clinic of a university affiliated teaching hospital.[br]Outcomes: The outcomes evaluated included early and late pregnancy losses, preterm delivery, birth weights and five neonatal complications.[br]Results: Of the 951 patients, whose data was available, 21 were antibody positive euthyroid patients (GpA), 376 had subclinical hypothyroidism (GpB), 98 had overt hypothyroidism (GpC), 2 had overt hyperthyroidism (GpD) and 31 had isolated hypothyroxemia (GpE). The rest 423 were included as the reference group (GpR). GpA, GpD and GpE did not show any statistical difference in outcomes when compared to the GpR. Subclinical hypothyroidism (GpB) was associated with significantly increased foetal loss, preterm delivery, increased low birth weight(LBW)rates and increased small for gestation age(SGA)rates, the odds ratios (95% CI) for these being 2.87(1.67-4.95), 1.91(1.06-3.44), 1.82(1.27-2.61) and 2.11(1.40-3.18) respectively. Overt hypothyroidism (GpC) was also associated significantly increased foetal loss, increased LBW rates, increased SGA rates and increased incidence of neonatal hyperbilirubinemia, the odds ratios (95% CI) for these being 2.54(1.17-5.51), 1.83(1.06-3.15), 2.12(1.15-3.89) and 3.14(1.57-6.25) respectively.[br]Conclusions: Subclinical hypothyroidism is associated with a similar magnitude of risk as overt hypothyroidism with regards to pregnancy loss and birth weights.[br][br](1) Panesar et al., Ann Clin Biochem 2001;38:329.[br][br]Nothing to Disclose: KA, VG, SA, TD, JJJ, KA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1080 49 134 SAT-13 PO32-01 Saturday 73 2012


74 ENDO12L_SAT-14 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Newly Diagnosed Hyperthyroidism in the Third Trimester of Pregnancy Manige Konig, Rana Malek, Eleanor Grimm Hutchens University of Maryland School of Medicine, Baltimore, MD Background: Hyperthyroidism in the third trimester of pregnancy is an uncommon phenomenon and should lead to further investigation. Choriocarcinoma is one of the possible causes of hyperthyroidism.[br]Clinical case: A 34 year old Asian female presented with increasing shortness of breath, weakness, hemoptysis to the hospital. Patient was 31 weeks gravid (G4P1A2). She had no other past medical history and had received most of her prenatal care in China.[br]Lab test results showed hyperthyroidism with a TSH of [lt] 0.02 uIU/ml (0.50-4.50 uIU/ml), free T4 of 4.8 ng/dL (0.7-1.8 ng/dL) and total T3 of 2.86 ng/ml (0.80-2.00 ng/ml).[br]She was started on methimazole 30 mg oral every 24 hours.[br]After 1 week of treatment her free T4 remained elevated with 3.4 ng/dl (0.7-1.8 ng/dl).[br]Further work up for possible autoimmune causes for hemoptysis, shortness of breath remained negative.[br]One week after admission, her respiratory status declined requiring intubation. CT-angio was negative for a pulmonary embolism but revealed multiple bilateral lung masses concerning for metastasis or granulomatous disease.[br]A b-HCG was obtained which showed a high level of 1,433,740 mIU/ml confirming the diagnosis of metastatic choriocarcinoma. The fetus was delivered emergently via C-section and survived without complications.[br]The surgical pathology report of the placenta confirmed the diagnosis of a choriocarcinoma.[br]The patient was started emergently on chemotherapy with EMA/CO (etoposide, methotrexate, actinomycin D, cyclophosphamide) and with a downwards trend in b-HCG the thyroid function normalized by day 8 of chemotherapy with a normal fT4 of 1.2 ng/dl (0.7-1.8ng/dl). At this point methimazole was discontinued.[br]Unfortunately the patient was pronounced brain dead after an acute bilateral parenchymal hemorrhage with left to right midline shift and ventricular entrapment.[br]Conclusion: This is a rare case in which a viable pregnancy co-existed with metastatic choriocarcinoma. HCG is composed of an a and b subunit. Hyperthyroidism can be induced via the homology of the a subunit of HCG to TSH; however, the lower potency of HCG to the TSH receptor requires HCG levels [gt] 100,000 mIU/mL to cause hyperthyroidism. The treatment is chemotherapy and methimazole until thyroid function tests normalize. Choriocarcinoma must be in the differential for all pregnant patients presenting with new hyperthyroidism in the third trimester.[br][br]Nothing to Disclose: MK, RM, EGH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 203 49 135 SAT-14 PO32-01 Saturday 74 2012


75 ENDO12L_SAT-15 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Re-Evaluation of Estrogen Receptor Expression in Endometriotic Cells Masao Izawa, Fuminori Taniguchi, Tomoiki Kiyama, Naoki Terakawa, Tasuku Harada Tottori University School of Medicine, Yonago, Japan; Tottori University School of Medicine, Yonago, Japan [bold]Background:[/bold] Endometriosis is an estrogen-dependent disease. The role of estrogen is obvious because the symptoms associated with endometriosis disappear after menopause, and the administration of GnRH agonists or progestin relieves the pelvic disease and the associated pain. A marked up-regulation of aromatase gene leads to a high estrogen environment in endometriotic tissues (1, 2). The urgent issue to be clarified is to understand the molecular basis of estrogen action. So far, a higher estrogen receptor [beta] (ER[beta]) and a lower estrogen receptor [alpha] (ER[alpha]) in endometriotic tissues have been documented to explain the pathophysiology of endometriosis. However, the recent discovery of multiple ER[beta] isoforms with distinct functions prompted us to re-evaluate the molecular basis of estrogen receptor (ER) expression in endometriotic tissues.[br][bold]Objective:[/bold] We re-evaluated the molecular basis of ER expression in stromal cells from chocolate cysts of patients with endometriosis.[br][bold]Patients:[/bold] The chocolate cyst lining in ovaries of patients with endometriosis was the source of endometriotic tissue. As control, endometrial tissues were obtained from uteri of cycling premenopausal women who had uterine leiomyoma. These patients had received no hormonal treatment before surgery. We obtained the informed consent from all patients.[br][bold]Methods:[/bold] Stromal cells were purified from endometriotic and endometrial tissues. ER[alpha] and ER[beta] mRNAs were assayed by real-time RT-PCR. Specific primer sets of unique 5[apos]-untraslated exons/exon2 in [italic]ESR1[/italic] were used for ER[alpha] promoter assay. Specific primer sets of unique 5[apos]-untraslated exons/exon1 in [italic]ESR2[/italic] were used for ER[beta] promoter assay.[br][bold]Results:[/bold] The expression of ER[alpha] mRNA in endometrial cells was 7-fold higher than that in endometriotic cells. A single cDNA sequence predicting a 66KDa ER[alpha] was amplified. 5[apos]-untranslated exons, C, D and F, were expressed. The expression of ER[beta] mRNA in endometriotic cells was higher than that in endometrial cells. Transcripts for ER[beta] isoforms ([beta]1, [beta]2, [beta]4 and [beta]5) were detectable. Among them, the ER[beta]5 was the most dominant. The expression was 4-fold higher than the ER[beta]1 in endometriotic cells. The 5[apos]-untranslated exon ON was expressed.[br][bold]Conclusion:[/bold] We re-evaluated the molecular basis of ER expression in endometriotic cells. Along with a marginal level of ER[alpha] mRNA, transcripts for 4 ER[beta] isoforms were demonstrated. These findings may provide a new facet in understanding the pathophysiological role of estrogen in endometriosis.[br][br]1. Izawa M, Harada T et al., Fertil Steril 2008; 89: 1390. 2. Izawa M, Taniguchi F et al., Fertil Steril 2011; 95: 33.[br][br]Nothing to Disclose: MI, FT, TK, NT, TH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1178 49 136 SAT-15 PO32-01 Saturday 75 2012


76 ENDO12L_SAT-16 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Estradiol Metabolites and Changes in Insulin Sensitivity during Oral Contraceptive Use Kai I Cheang, Xia Xu, Timothy D Veenstra, John E Nestler Virginia Commonwealth University, Richmond, VA; National Cancer Institute, Frederick, MD; Virginia Commonwealth University, Richmond, VA BACKGROUND: Deterioration in insulin sensitivity has been reported in some women taking combined hormonal oral contraceptives (OC). Mouse models suggest the estradiol metabolites 2-hydroxyestradiol (2OHE2) and 2-hydroxyestrone (2OHE1) regulate glucose metabolism. We prospectively evaluated changes in estradiol metabolites during OC use and their effect on insulin sensitivity.[br]METHODS: In 25 women during administration of ethinyl estradiol 35mcg and norgestimate 0.18/0.215/0.25 mg, insulin sensitivity (Si) was evaluated by frequently sampled IV glucose tolerance test, and serum endogenous estrogens and metabolites were quantified by LC-MS2 at baseline, 3 and 6 months. Pearson[apos]s correlations were used to evaluate associations between [Delta]Si and [Delta]estrogen metabolites. Subjects were also stratified into those whose Si worsened and those whose Si remained unchanged during the 6 months. Trends of 2OHE1 and 2OHE2 levels during 6 months of OC use between the 2 groups were evaluated by repeated measures analyses.[br]RESULTS: Ratios of 2OHE1/Total estrogens (-2.5[plusmn]2.0, p=0.003) and 2OHE2/Total estrogens (-0.11[plusmn]0.05, p=0.002) decreased during OC use. Although Si did not change significantly (+0.62[plusmn]0.4 min-1/mu/L, p=0.4198) overall after 6 months of OC, substantial inter-individual variability exist, and 50% of subjects had worsened Si. [Delta]2OHE2 (p=0.043) and [Delta]2OHE2/Total estrogens (p=0.045) were directly correlated with [Delta]Si. When participants were divided into whether their Si deteriorated or remained unchanged during the 6 months, divergent patterns of 2OHE2 were observed. Women whose Si deteriorated exhibited a more pronounced decrease in 2OHE2/Total than women whose Si remained changed (-0.38[plusmn] 0.29 vs. -0.08 [plusmn] 0.30 [x10-2], p=0.0166).[br]CONCLUSIONS: Changes in endogenous estradiol metabolism may determine OC[apos]s effects on insulin sensitivity.[br][br]Sources of Research Support: NIH K23HD049454; UL1RR031990; American Heart Association 11GRNT7730026 awarded to KIC.[br][br]Nothing to Disclose: KIC, XX, TDV, JEN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1303 49 137 SAT-16 PO32-01 Saturday 76 2012


77 ENDO12L_SAT-17 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Use of a Quality of Life (QOL) Menopause Rating Scale (MRS) To Define Hormone and Aging Symptoms during the Menopause and Post-Menopause (Geripause) Bernard A Eskin, Jay Tank, Owen Montgomery, Christopher Sell Drexel University College of Medicine, Philadelphia, PA; Drexel University College of Medicine, Philadelphia, PA [bold]Introduction.[/bold] The menopausal transitions are distinct in th lives of women, with certain common physiologic events thaat occur, related to the imbalance of the hypothalaamic-pituitary-ovarian axis. The symptoms experienced are myriad, and include all domains of a woman[apos]s life - psychological, physiological, physical, and sexual. While each woman is unique in terms of the specific symptoms experienced, including their severity and frequency, it is evident that they affect their lives to a great extent. They can lead to a substantial decrease of the QOL. The MRS was developed in response to the lack of standardized scales to measure the aseverity of aging symptoms and their impact on the health-related quality of QOL.[br][bold]Materials and Methods.[/bold] 60 women (20 in each group 40-49, 50-64, 65+) were randomly recruited. The patients were consented on the research study, given the MRS questionnaire (MQ) to fill out on their own, and a blood sample was taken for hormonal assays. The MQ has been standardized to include the 11 most common symptoms a woman might experience during the pre-menopausal, menopausal, and post-menopausal transition period. It consists of a list of 11 symptoms grouped into 3 domains: psychological, physiological, somato-vegetative, and urogenital. The respondents havee 5 choices for each symptom listed: no symptoms, mild, moderate, marked, and severe, with a score of 0-4, respectively. The composite scores for each of the dimensions are based on adding up the scores of these symptoms.[br][bold]Results.[/bold] The results from the MQ were stratified but stable in all age groups. The changes in symptoms are determined by many factors. In this study we looked at the results from the MQ to more clearly delineate the effects of aging versus hormone loss. They showed a significant difference between the pre-menopausal and early post-menopausal groups, particularly the psychological symptoms. Several theoies have been suggested for random changes seen in the statistics, but there is an high degree of varaibility.[br][bold]Conclusion.[/bold] The MRS questionnaire has been shown to have substantial validity and reliability, and should be considered as a potential diagnostic tool. One advantage of using the MRS is its complete reliance on the patient for the answers, leading to more accurate representation of the most concerning symptoms. This can lead to more defined therapeutic modalities that will benefit the patient.[br][br]Nothing to Disclose: BAE, JT, OM, CS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1548 49 138 SAT-17 PO32-01 Saturday 77 2012


78 ENDO12L_SAT-18 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) The Effect of a Tissue-Specific Estrogen Complex (TSEC) on Normal Mammary Gland and Breast Cancer Xenografts in Mice: Rationale for Use of a Tissue-Selective Estrogen Complex Yan Song, Wei Yue, Jiping Wang, Richard J Santen University of Virginia, Charlottesville, VA The Women[apos]s Health Initiative (WHI) studies showed that the combination of an estrogen plus a progestin increased the risk of breast cancer in postmenopausal women while estrogen alone did not. As a novel means of eliminating the need for a progestin as menopausal hormone therapy (MHT), use of a tissue selective estrogen complex (TSEC) has been proposed. The most studied TSEC in women involves the pairing of a SERM, bazedoxifene (BZA) with conjugated estrogens (CE). Our hypothesis is that this combination may prevent the growth of occult breast cancers in women. To systematically examine the effects of this TSEC on breast, we quantified its effects on mouse mammary gland and occult human breast cancer xenografts. We initially used computerized morphometry to examine the effects of estradiol (E[sub]2[/sub]) and CE alone or in combination with BZA on ductal development in immature castrate mice. BZA completely blocked CE or E[sub]2[/sub] stimulated ductal and terminal end bud growth (P[lt]0.05). We then examined human breast tumor proliferation. Occult, undiagnosed breast cancer is present at autopsy in 7% of women between the ages of 40 and 80 (Santen et al JCEM 95:S1-66, 2010). Using a MCF-7 mouse xenograft model to mimic occult tumors in women, we demonstrated that plasma levels of 80 pg/ml of E[sub]2[/sub] (estrogen clamp method) stimulated growth of small tumors. This effect was completely blocked with BZA (2 mg/kg) (p[lt]0.05). BZA also inhibited E[sub]2[/sub] stimulation of PR, pS2, cMyC and AREG; the enhancement of Ki67 and PCNA as proliferation markers; and the anti-apoptotic effect of estrogen. CE was much less potent than E[sub]2[/sub] on reducing apoptosis and on stimulation of Ki67 expression. In addition, gene expression analysis showed less stimulation of proliferation related genes (AREG, cyclin D1, and cMyc) by CE compared to E[sub]2[/sub] but all effects were blocked by BZA. Surprisingly CE alone did not stimulate tumor growth but did cause a 6-fold increase in uterine weight. This uterine weight bioassay confirmed that CE were effectively absorbed, de-conjugated in vivo, and capable of exerting an estrogenic effect on uterus but not on breast cancer. In summary, these results demonstrate that BZA can abrogate the growth of E[sub]2[/sub] stimulated occult human breast tumors. The estrogenic effects of CE on cellular markers could be blocked by BZA. These data support our hypothesis that the CE/BZA TSEC exerts anti-estrogenic effects on benign and malignant breast tissue.[br][br]Santen et al., JCEM 2010; 95:S1.[br][br]Sources of Research Support: Pfizer Inc.[br][br]Disclosures: RJS: Advisory Group Member, Pfizer, Inc., Novo Nordisk; Consultant, Teva. Nothing to Disclose: YS, WY, JW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1584 49 139 SAT-18 PO32-01 Saturday 78 2012


79 ENDO12L_SAT-19 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Development and Characterization of a Hyperglycosylated VEGF Analogue That Is Superior to Recombinant VEGF for Stimulating Angiogenesis during Ovarian Folliculogenesis in Rats Rhonda K Trousdale, Martin A Julius, Susan V Pollak, Joyce W Lustbader Columbia University, New York, NY; Columbia University, New York, NY; Harlem Hospital, New York, NY Physiological post-natal angiogenesis occurs cyclically in reproductive age women. Within the ovary, follicles rapidly expand during folliculogenesis. The independent vascular network surrounding each individual follicle must also expand rapidly to provide adequate oxygen, food, and nutrients to the growing structure. As women age, poor ovarian follicle vascular development is associated with reduced fertility.[br]A unique aspect of evaluating angiogenesis in rat ovarian follicles is the ability to accurately quantify vascular area for a well-defined structure as opposed to only evaluating vascular density. We previously reported that during non-stimulated folliculogenesis, as follicles increase in size the vasculature develops in a reproducible ratio(1). However, treatment with a long-acting follicle stimulating hormone analogue (FSH-CTP) promotes rapid follicle development with a lag in vascular development. Co-administration of FSH-CTP with recombinant vascular endothelial growth factor (rVEGF) did not enhance enhance follicle vascular development(1). We hypothesized rVEGF therapy was ineffective due to the inherently short half-life of VEGF(30 min).[br]We developed a hyper-glycosylated VEGF protein(VC). Hyper-glycosylation increased the half-life 3-fold to 80 min with decline to baseline at 200 min.[br]To evaluate the effect of VEGF monotherapy on follicle vasculature, 21 day female Sprague Dawley rats were given saline; rVEGF(1[mu]g); or VC(1[mu]g) at 0 and 24 hours. At 48 hours ovaries were evaluated. There was no difference in follicle vasculature development between these 3 groups.[br]However, when rats were stressed by administration of FSH-CTP, co-administration with VC significantly enhanced all parameters measured including mean antral follicle number, mean follicle size, and mean follicle vasculature. Furthermore, VC co-administration restored the ratio of vasculature to inner area back to the level of non-FSH-CTP treated rats.[br]This study demonstrates hyperglycosylation of VEGF-A[sub]165[/sub] does not interfere with protein activity and enhances the half-life without loss of biological activity. While administration of rVEGF during the stress of FSH-CTP ovarian hyperstimulation was unable to improve vasculogenesis, VC co-administered with FSH-CTP was able to enhance ovarian follicle vascular development leading to an increase in the number of large, healthy antral follicles. Short-term administration of VC during [italic]in vitro[/italic] fertilization may improve fertility in older women.[br][br](1)Trousdale RK et al., Endocrinology 2007; 148:1296.[br][br]Sources of Research Support: Juvenile Diabetes Foundation Innovative Grant; NIH Grant DK063224; NIH Grant HD055126 awarded to JWL.[br][br]Nothing to Disclose: RKT, MAJ, SVP, JWL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1658 49 140 SAT-19 PO32-01 Saturday 79 2012


80 ENDO12L_SAT-20 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Development of a Well-Characterized PAPP-A2 ELISA To Measure Bioactive PAPP-A2 in Maternal Biological Fluids Bhanu Kalra, Ajay Kumar, Anthony Tran, Claus Oxvig, Shivani Shah AnshLabs, Webster, TX; University of Aarhus, Gustav Wieds, Denmark Pregnancy-associated plasma protein-A2 (PAPP-A2) is a novel metalloproteinase identified as a homolog of PAPP-A in the metzincin superfamily of pappalysins. PAPP-A2 shares 46% sequence identity with PAPP-A. PAPP-A2 is a noncovalently linked dimer of two 220-kDa subunits. It exhibits roboust proteolytic activity against IGFBP-5 and possibly also IGFBP-3. PAPP-A2 is expressed in a wide range of tissues and is abundant in placental syncytiotrophoblasts and the pregnant uterus. The physiological importance of PAPP-A2 is not fully known.[br][bold]Methodology:[/bold] We have developed a well characterized two-step sandwich-type enzymatic microplate ELISA to measure PAPP-A2 levels in the maternal serum and other biological fluids. The assay measures PAPP-A2 in 50 [micro]L of sample (diluted 20 folds in sample diluent) against recombinant PAPP-A2 calibrators (0.1-10 ng/mL). The antibody pair used in the PAPP-A2 ELISA measures bioactive PAPP-A2 and does not detect PAPP-A2 missing the C-terminal region (residues 1397-1791), dimeric PAPP-A, PAPP-A proMBP complex and MMP-9.[br][bold]Validation [/bold]Total imprecision calculated on 4 samples over 12 runs, 4 replicates per run, using CLSI EP5-A guidelines was 4.7% at 1.03ng/mL, 4.% at 1.8ng/mL, 4.6% at 2.6ng/mL and 4.4% at 3.13ng/mL. The limit of detection calculated using six serum samples in the range of 0.091-3.085ng/mL over 12 runs is 0.071ng/mL.[br]The functional sensitivity of the assay at 20% CV was 0.08ng/mL. Dilution studies showed an average recovery of 100-110%. The median PAPP-A2 value on a first trimester samples (n=65) and second trimester samples (n=65) were 30.76ng/mL and 42.14ng/mL, respectively. When potential interferents (hemoglobin, triglycerides and bilirubin) were added at two times their physiological concentration, PAPP-A2 concentrations were within [plusmn]10% of the control.[br][bold]Conclusions:[/bold] A quantitative, robust and fully characterized microplate PAPP-A2 ELISA has been developed to measure PAPP-A2 in maternal serum. The approximate median PAPP-A2 levels found in a first and second trimester maternal serum can be measured with [lt] 5 % CV using this assay. The performance of the assay is acceptable for investigation of clinical utility in a variety of maternity related disorder.[br]* For Research Use Only. Not for use in diagnostic procedures.[br][br]Nothing to Disclose: BK, AK, AT, CO, SS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1690 49 141 SAT-20 PO32-01 Saturday 80 2012


81 ENDO12L_SAT-21 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Evaluation of Vascular Aging Using Myometrial Artery Calcification and Intima Thickness Sarah Catherine Hessler, Gerson Weiss, Debra S Heller, Peter G McGovern, Catherine Susan Delia, Laura T Goldsmith UMDNJ-New Jersey Medical School, Newark, NJ Although the incidence of cardiovascular disease increases with advancing age in women, studies have not successfully distinguished the effects of changing hormonal status on vascular aging from the effects of chronological age. Currently, models of human vascular aging that are generally used involve either imaging techniques or invasive sampling to assess markers of vascular aging, such as calcification and intimal hyperplasia. Occurrence of these markers has been shown to increase with advancing age and correlates with the presence of coronary heart disease. In the United States, greater than 500,000 hysterectomies are performed each year. No studies have demonstrated whether myometrial vessels can be used for the study of vascular aging. We hypothesized that myometrial arteries are a useful model for assessment of markers of vascular aging and that occurrence of these markers increases with age.[br]To test this hypothesis, myometrial arteries were assessed for calcifications utilizing hematoxylin and eosin-stained slides, and for intimal thickness utilizing orcein-stained slides for elastin. Tissues from 45-70 year old subjects who had a benign indication for hysterectomy were studied. Exclusion criteria were diagnosis of hypertension or diabetes, or treatment with sex steroids within 3 months prior to surgery. Subjects were categorized as premenopausal if they had their last menstrual period within 6 months. Calcifications were determined to be present or absent in tissues from all subjects (n=50). Intimal thickness was assessed in premenopausal women (n=16). For each subject, to assess intimal thickness, three myometrial arteries at least 0.5mm diameter were scored in a blinded manner according to the percent occlusion of the vessel lumen using the following scale: 0, less than 5%; 1, 5-20%; 2, 21-50%; or 3, greater than 50%. Data were assessed by linear regression.[br]In premenopausal women age 45-55 years, a significant positive relationship between age and intimal thickness was observed (p =0.035), intimal thickness increased with increasing age. Myometrial artery calcifications were infrequently observed, only in 2 post-menopausal subjects.[br]Myometrial arteries in hysterectomy specimens are an appropriate model for the study of vascular aging. The abundance of hysterectomy specimens may allow better differentiation of the effects of age and hormonal status on vascular aging.[br][br]Nothing to Disclose: SCH, GW, DSH, PGM, CSD, LTG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1697 49 142 SAT-21 PO32-01 Saturday 81 2012


82 ENDO12L_SAT-22 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Androstenediol May Interact with Estradiol To Modulate Vasomotor Symptoms during the Menopausal Transition Sybil Crawford, Daniel S McConnell, Sara R Pittenger, Marcelle I Cedars, Frank Z Stanczyk, Ellen B Gold, Nancy A Gee, Bill L Lasley University of Massachusetts, Worcester, MA; University of Michigan, Ann Arbor, MI; University of California, San Francisco, CA; University of Southern California, Los Angeles, CA; University of California, Davis, CA; University of California, Davis, CA; University of California, Davis, CA Background: During the menopausal transition (MT) circulating estradiol (E2) concentrations decrease modestly in all women while circulating adrenal delta five steroids can increase up to ten fold. Androstenediol (Adiol) ranges in concentration from 100 to 1,000 pg/mL at or near the final menstrual period (FMP), while circulating E2 levels may decline to 10 pg/mL or less. E2 is primarily an estrogen receptor (ER) alpha ligand while Adiol can activate both the ER alpha and ER beta receptor-mediated pathways and possesses both estrogenic and androgenic properties. Circulating E2 concentrations are negatively related to the frequency of vasomotor symptoms (VMS) and intervention with E2 is effective in attenuating hot flushes. However, direct measurements of circulating E2 do not accurately predict the occurrence or severity of hot flushes such that inter-woman differences in the VMS experience have not been adequately explained. The wider range of circulating Adiol, compared to E2, may help explain between women differences in VMS symptoms and the lack of correlation with circulating E2.[br]Aim: To characterize the relationship of E2, Adiol and the E2/Adiol ratio in relation to the occurrence and frequency of hot flushes in women during the MT.[br]Methods: Logistic regression analysis was conducted for mid-aged women (n=142 women, 998 observations) during the MT for VMS frequency (infrequent = 1-5 days in the past 2-weeks) and frequent (6+ days in the past two weeks) versus none.[br]Results: Circulating E2 was negatively related to VMS, in the setting of both low Adiol (Odds ratio (OR) =0.37, 95% CI 0.24 [ndash] 0.59, p[lt]0.0001) and high Adiol (OR=0.63, 95% Confidence interval (CI) 0.39 [ndash] 1.03, p=0.065). Adiol was negatively related to VMS at low E2 (OR 0.72, 95% CI 0.39 [ndash] 1.33, p=0.296) but positively related to VMS at high E2 (OR 1.22, 95% CI 0.59 [ndash] 2.50, p=0.590), although the interaction was not statistically significant (p=0.137).[br]Conclusion: These data suggest that while VMS frequency has some relation to circulating E2 concentrations, the protective effect of E2 on VMS may be modulated by the antagonistic effects of the relatively higher circulating Adiol levels.[br][br]Sources of Research Support: The Study of Women[apos]s Health Across the Nation (SWAN) has grant support from the National Institutes of Health (NIH), DHHS, through the National Institute on Aging (NIA), the National Institute of Nursing Research (NINR) and the NIH Office of Research on Women[apos]s Health (ORWH) (Grants NR004061; AG012505, AG012535, AG012531, AG012539, AG012546, AG012553, AG012554, AG012495, AG017719). The content of this abstract is solely the responsibility of the authors and does not necessarily represent the official views of the NIA, NINR, ORWH or the NIH.[br][br]Nothing to Disclose: SC, DSM, SRP, MIC, FZS, EBG, NAG, BLL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1738 49 143 SAT-22 PO32-01 Saturday 82 2012


83 ENDO12L_SAT-23 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Development of a Fully Characterized Total PAPP-A Chemiluminescence Assay for Maternal Serum Evaluation* Ajay Kumar, Bhanu Kalra, Amita S Patel, Claus Oxvig AnshLabs, Webster, TX; University of Aarhus, Gustav Wieds, Denmark Maternal serum concentrations of pregnancy-associated plasma protein A (PAPP-A, EC 3.4.24.79, papalysin-1) are used to predict occurrence of Down syndrome. During pregnancy, PAPP-A is produced in high concentrations by the trophoblast and released into maternal circulation. In pregnancy, PAPP-A primarily circulates as 500-kDa heterotetrameric 2:2 complex with the proform of eosinophil major basic protein (proMBP), which inhibits the proteolytic activity of PAPP-A. Dimeric PAPP-A is the only active form and proteolyse IGFBP-4 and IGFBP-5. Significant amounts of active PAPP-A is reported at gestational ages between seven and thirteen weeks.[br][bold]Methodology:[/bold] We have developed a well characterized two-step sandwich-type enzymatic microplate glow based chemiluminiscence assay to measure PAPP-A levels in the maternal serum. The assay measures PAPP-A in 50 [micro]L of serum sample (diluted 100 folds in sample diluent) against heterotetrameric PAPP-A calibrators (25-5000 ng/mL). The antibody pair used in the assay measures dimeric PAPP-A and PAPP-A/proMBP complex in almost equal concentration and does not cross-react with proMBP, PAPP-A2 and MMP-9 at twice the physiological concentrations.[br][bold]Validation:[/bold] All analytical results are reported in ng/mL. One ng/mL of htPAPP-A = 2.56 [mu]IU/mL. Total imprecision calculated on 3 samples over 10 runs, 4 replicates per run, using CLSI EP5-A guidelines was 3.1% at 249ng/mL, 4.3% at 476ng/mL and 5.7% at 732.5ng/mL. The limit of detection calculated using six serum samples in the range of of 3.95-159.15ng/mL over 10 runs is 7.73ng/mL. The functional sensitivity of the assay at 15% CV was 11.65ng/ml. Dilution studies showed an average recovery of 96-109%. The median PAPP-A value on a first trimester samples (n=50) was 643.68ng/mL. When potential interferents (hemoglobin, triglycerides and bilirubin) were added at two times their physiological concentration, PAPP-A concentrations were within [plusmn]10% of the control.[br][bold]Conclusions:[/bold] A quantitative, robust and fully characterized microplate PAPP-A chemiluminiscence assay has been developed to measure PAPP-A in maternal serum. The approximate median PAPP-A levels found in a first trimester maternal serum can be measured with [lt] 7 % CV using this assay. The performance of the assay is acceptable for investigation of clinical utility in a variety of maternity related disorder.[br]* For Research Use Only. Not for use in diagnostic procedures.[br][br]Nothing to Disclose: AK, BK, ASP, CO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1778 49 144 SAT-23 PO32-01 Saturday 83 2012


84 ENDO12L_SAT-24 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Pilot Study on Short-Term Effects of Aromatase Inhibitor Therapy on Cognitive Function in Postmenopausal Women with Breast Cancer Shalini Dabbadi, Paula Gendreau, Susan Tannenbaum, Susan Rabinowe, Richard Kaplan, Faryal Mirza University of Connecticut Health Center, Farmington, CT; University of Connecticut Health Center, Farmington, CT; University of Connecticut Health Center, Farmington, CT; Saint Francis Hospital and Medical Center, Hartford, CT BACKGROUND: Mild cognitive impairment (MCI) has been reported in cross-sectional studies of postmenopausal women with breast cancer receiving aromatase inhibitors (AIs). A potential underlying mechanism for this finding may be the reduction in the formation of female sex hormones from precursor androgens with AI therapy. Estrogen receptors have been identified in several areas of the brain important in cognitive performance including prefrontal cortex (short-term memory), the hippocampus (learning and storage) and the amygdala (modulation of memory consolidation).[br]OBJECTIVES: To determine the effect of short term therapy with AIs, letrozole and anastrozole, on cognitive function in postmenopausal women with breast cancer.[br]METHODS: Comprehensive neurocognitive testing was performed in postmenopausal women initiating therapy with anastrozole or letrozole for breast cancer. Subjects were evaluated at baseline and at 12 weeks after initiating therapy using the Rey Auditory Verbal Learning Test, Benton Visual Retention Test, Trail Making Test, the Stroop Color and Word Test and the Controlled Oral Word Association Test (COWAT). The Cognitive Failures Questionnaire and Beck Depression Inventory were also administered to assess perceived cognitive deficits and symptoms of depression. All tests were administered according to standardized published procedures.[br]RESULTS: 28 postmenopausal women with mean age of 59 [plusmn] 6 years completed the study. A significant decline in immediate memory (t = 2.6, df = 27, p [lt]0.02) and delayed memory (t = 2.3, df = 27, p [lt]0.03) was observed while the COWAT scores improved significantly (t = -2.8, df = 27, p [lt]0.01). There were no significant changes on the Stroop or Trail Making Tests (attention, executive functioning, non-verbal memory and processing speed for color and word). There were also no significant differences in self-reported depression and anxiety scores. Greater estrogen decline correlated with lower delayed memory scores on follow-up (r = -0.30), although it did not reach statistical significance.[br]CONCLUSION: Recall memory for a word list declined in the first three months of therapy with AIs in women with breast cancer suggesting the role of estrogen depletion in MCI. Our results suggest the need to further explore the assessment of short and long term cognitive function with adjuvant endocrine treatment of breast cancer as these findings may temporarily or permanently impact the quality of life in this population.[br][br]Sources of Research Support: Connecticut Breast Health Initiative. General Clinical Research Center, University of Connecticut Health Center.[br][br]Nothing to Disclose: SD, PG, ST, SR, RK, FM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1792 49 145 SAT-24 PO32-01 Saturday 84 2012


85 ENDO12L_SAT-25 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) SHBG, but Not Estrogen and Testosterone, Is Beneficially Associated with Adipokines in Postmenopausal Women Roksana Karim, Wendy J Mack, Chun-Ju Chien, Howard N Hodis, Frank Z Stanczyk University of Southern California, Los Angeles, CA; University of Southern California, Los Angeles, CA; University of Southern California, Los Angeles, CA; University of Southern California, Los Angeles, CA [underline]Background:[/underline] Adipose tissue is a major source of estrogens in postmenopausal women. Little is known about the association of estrogens and adipokines in this population. Till date, only one cross-sectional study evaluated the association of adiponectin with sex hormones and sex hormone binding globulin (SHBG) in postmenopausal women. The associations of leptin, resistin, and ghrelin with sex hormones and SHBG are largely unknown in this population.[br][underline]Methods:[/underline] A cross-sectional study in 643 postmenopausal women not taking any hormone therapy. Using stored frozen samples from the baseline visit, serum concentrations of estrone (E1), total estradiol (E2), and total testosterone (T) were measured by validated extraction/chromatographic RIAs; SHBG was measured by direct chemiluminescent immunoassay. Free estradiol (FE2) and free testosterone (FT) levels were calculated. Serum leptin, adiponectin, resistin, and ghrelin concentrations were quantified by RIA. Concentrations of all the analytes were tested for association with time since menopause, BMI, and waist-hip ratio (WHR), using linear regression models. Linear regression models were also used to evaluate the associations of sex hormones and SHBG (dependent variables) with adipokines and ghrelin (independent variables) adjusted for age, race, and BMI.[br][underline]Results:[/underline] The mean (SD) age of the women was 60 (6.9) years, the 68.5% white, and 58% were [gt]10 years from menopause. SHBG, adiponectin, and leptin concentrations but no sex hormones were significantly greater among women [gt]10 years (all p-values [lt]0.03). E1, E2, FE2, and FT were significantly inversely associated with adiponectin and leptin (all p[lt]0.03) adjusted for age, race, and BMI. Only estrogens were significantly inversely associated with ghrelin (all p[lt]0.01). On the contrary, SHBG levels were significantly positively associated with adiponectin and inversely associated with leptin.[br][underline]Conclusions:[/underline] Low adiponectin and ghrelin and high leptin levels are reportedly associated with obesity and increased risk of cardiovascular disease (CVD) whereas high SHBG levels are protective. In that context, our data indicate adverse association of estrogens and testosterone with adipokines and ghrelin. On the contrary, high SHBG levels are beneficially associated with adipokines. Further research is warranted to understand the underlying mechanisms of interaction between adipokines and sex hormones pertaining to CVD.[br][br]Sources of Research Support: Robert E. and May R. Wright Foundation Award, University of Southern California.[br][br]Nothing to Disclose: RK, WJM, C-JC, HNH, FZS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1882 49 146 SAT-25 PO32-01 Saturday 85 2012


86 ENDO12L_SAT-26 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Effects of Conjugated Estrogens and Estradiol on Breast Cancer Growth [italic]In Vitro[/italic] and Antagonism by Bazedoxifene: Rationale for Use of Tissue-Selective Estrogen Complexes (TSECs) Yan Song, Wei Yue, Jiping Wang, Richard J Santen University of Virginia, Charlottesville, VA The Women[apos]s Health Initiative (WHI) trial of menopausal hormone therapy (MHT) demonstrated an increased incidence of breast cancer in women receiving Premarin (conjugated estrogens, CE) plus the progestin, medroxyprogesterone acetate, but strikingly, not with Premarin alone. The use of a TSEC (tissue selective estrogen complex) has been proposed as a novel MHT strategy to eliminate the requirement for a progestin. The first TSEC in clinical development, which pairs the SERM, bazedoxifene (BZA) with conjugated estrogens improves hot flashes, vaginal atrophy, and bone density without stimulation of uterine or benign breast tissue. We posed the question whether this TSEC would exert anti-estrogenic effects on breast and potentially prevent breast cancer. Seven % of women from ages 40-80 harbor occult, undiagnosed breast cancer at autopsy (Santen et al., JCEM 95:S1-66, 2010) and a TSEC might inhibit growth of these tumors. To address this issue, we initially compared estradiol (E[sub]2[/sub]) and CE alone on proliferation and apoptosis in MCF-7 breast cancer cells. CE dose-dependently stimulated growth of MCF-7 cells at a peak concentration (10[sup]-9[/sup] M) ten-fold higher than required for E[sub]2[/sub] (10[sup]-10[/sup] M). We found that both CE and E[sub]2[/sub] alone increased DNA synthesis and reduced apoptosis at similar concentrations and BZA (10[sup]-8[/sup] M) completely blocked these effects. We then used western blot analysis to evaluate several key protein markers linked to proliferation and apoptosis. Both CE and E[sub]2[/sub] significantly stimulated phosphorylation of MAP kinase, Akt, and p70S6K as proliferation markers and up-regulated survivin, Bcl-2, and XIAP as anti-apoptotic factors. These effects of CE and E[sub]2[/sub] could be completely blocked by BZA. Gene expression studies demonstrated that CE and E[sub]2[/sub] were equally potent on expression of cMyc, pS2, and WISP whereas the stimulatory effect of CE on PR and AREG expression was weaker than for E[sub]2[/sub]. BZA effectively blocked each of these effects and alone, demonstrated no estrogen agonistic effects when used under estrogen deprived conditions. In summary, our results indicate that the stimulatory effects of E[sub]2[/sub] or CE on breast tumor cells could be completely abrogated by BZA at concentrations equivalent to those used in women. These studies imply that the CE/BZA TSEC exerts anti-estrogenic effects on breast cancer and might block the growth of occult, undiagnosed breast tumors in postmenopausal women, resulting in a reduction of diagnosed breast tumors.[br][br](1) Santen et al., JCEM 2010; 95:S1.[br][br]Sources of Research Support: Pfizer, Inc.[br][br]Disclosures: RJS: Advisory Group Member, Pfizer, Inc., Novo Nordisk; Consultant, Teva. Nothing to Disclose: YS, WY, JW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2006 49 147 SAT-26 PO32-01 Saturday 86 2012


87 ENDO12L_SAT-27 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Oxytocin Activates NF-[kappa]B and MAPK Pathways in Human Amnion Sung H Kim, Andrew Blanks, Steve Thornton, Mark R Johnson, Phillip R Bennett, Vasso Terzidou Imperial College, London, UK; Warwick Medical School, Coventry, UK Human labour is associated with an increase in prostaglandin (PG) and inflammatory cytokine synthesis within the fetal membranes. Human amnion is known to be one of the major sources of PG production. Incubation of pre-labour amnion cells with oxytocin (OT) results in a marked increase in PGE[sub]2[/sub] synthesis and upregulation of COX-2 and cPLA[sub]2[/sub], key enzymes involved in PGE[sub]2[/sub] synthesis. Our study has identified a novel role for OT in activating NF-kappaB in human amnion and has explored the regulatory mechanisms involved in OT-mediated COX-2 expression in human amnion.[br]Primary amnion cell cultures were established from patients having elective pre-labour caesarean section at term. We found that OT treatment results in nuclear translocation of p65, but not of p50 or RelB NF-kappaB subunits. OT activates p65 and IKK[alpha]/[beta] resulting in degradation of IkappaB[alpha] which could be inhibited using an IKK[beta] inhibitor (TPCA-1).[br]Treatment with OT upregulated mRNA expression of several NF-kappaB regulated genes, including IL-8, CCL2, CCL5 and COX-2 (p[lt]0.05, ANOVA). siRNA mediated knockdown of the p65 NF-kappaB subunit resulted in a significant suppression of OT induced COX-2 upregulation. Pre-treatment with IKK[beta] inhibitor (TPCA-1) also significantly decreased the upregulation of COX-2 by OT (p[lt]0.05, ANOVA).[br]We found that OT activates ERK1/2 and p38 kinase, but not JNK. Inhibition of ERK1/2 and p38 resulted in suppression of OT induced NF-kappaB activation and COX-2 upregulation. Inhibition of G[sub]i[/sub] protein by pertussis toxin (PTX) led to inhibition of OT induced phosphorylation of p65, ERK1/2 and p38, and suppressed OT induced COX-2 and p-cPLA[sub]2[/sub] upregulation.[br]We have demonstrated that OTR couples with G[sub]i[/sub] to activate NF-kappaB in human amnion by a novel mechanism, which requires both IKK and MAPK activation. Our findings suggest a potential further mechanism by which OTR-antagonists and NF-kappaB inhibitors might act in the management of preterm labour.[br][br]Sources of Research Support: Action Medical Research Grant 2010-2013.[br][br]Nothing to Disclose: SHK, AB, ST, MRJ, PRB, VT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2052 49 148 SAT-27 PO32-01 Saturday 87 2012


88 ENDO12L_SAT-28 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Development of Ovarian Cultures to Rapidly Test Chemotherapeutic Fertotoxicity [italic]In Vitro[/italic] and [italic]In Vivo[/italic] So-Youn Kim, Marilia Cordeiro, Vanida Ann Serna, Jessina Tomas, Lizbeth Gutierrez, Takeshi Kurita, Teresa K Woodruff Northwestern University, Chicago, IL Earlier detection of cancer, coupled with new and improved treatment options, has increased the number of survivors of cancer, especially those who are diagnosed as children. As the number of young cancer survivors increases, so does the concern for their quality of life post-treatment. Unfortunately, off-target effects of radiation and chemotherapies can have unexpected and lasting consequences. For young women, this is particularly worrisome, as the immature oocytes that comprise the ovarian reserve are particularly vulnerable to the effects of radiation and drugs. Our group has developed in vitro ovarian culture and isolated ovarian follicle culture systems in order to advance our understanding of follicle development and the effects of cancer treatments on the ovary, with the goal of finding new ways to restore fertility to young cancer survivors who have been rendered sterile by their cancer treatment. A recent study suggested that the tyrosine kinase inhibitor Gleevec[reg] (imatinib mesylate) protects oocytes from cisplatin-induced cell death when immature animals were treated. Options for preserving fertility in young women prior to cancer treatment are investigational and involve removal of an ovary; even in the face of certain sterility, this can be a difficult decision for young patients and their parents. Experiments aimed at more rapidly testing chemotherapies as well as identifying neo-adjuvant fertoprotective agents can be more rapidly advanced by the deployment of the in vitro follicle culture system and xenograph model described here. To test our in vitro culture system and the xenograph transplant model, we first treated ovaries with Gleevec[reg] in vitro. Here we show that Gleevec[reg] rescues primordial follicles by cisplatin in cultured mouse ovaries by blocking an apoptotic signaling pathway including Bax, cPARP and Caspase 3. Further, we show that the xenograph model recapitulates the in vitro results and extend our analysis to monitor rH2AX foci. Importantly, Gleevec[reg] decreases the number of young follicles containing rH2AX foci inducing DNA damage. These studies show that we have two model systems in which drug testing can occur rapidly, and aid our identification of fertoprotective agents in the future.[br][br]Sources of Research Support: NIH grant(HD041857).[br][br]Nothing to Disclose: S-YK, MC, VAS, JT, LG, TK, TKW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2068 49 149 SAT-28 PO32-01 Saturday 88 2012


89 ENDO12L_SAT-29 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Effects of Bazedoxifene/Conjugated Estrogens on Coagulation Parameters in a 1-Year, Randomized, Placebo- and Active-Controlled, Phase 3 Trial of Postmenopausal Women Sebastian Mirkin, Kelly A Ryan, John R Thompson, Arkadi A Chines Pfizer Inc, Collegeville, PA [bold]Objective:[/bold] In phase 3 clinical trials, bazedoxifene (BZA)/conjugated estrogens (CE) demonstrated efficacy in treating menopausal symptoms and preventing osteoporosis in postmenopausal women with a favorable safety/tolerability profile, including no increase in the risk of coronary heart disease or stroke. The risk of venous thromboembolism for BZA/CE appears to be similar to that for CE and BZA alone. The effects of BZA/CE on coagulation parameters were evaluated in the Selective estrogens, Menopause, And Response to Therapy (SMART)-5 trial.[br][bold]Methods:[/bold] The SMART-5 trial was a 1-year, randomized, double-blind, placebo (PBO)- and active-controlled, phase 3 study in non-hysterectomized, postmenopausal women aged 40 to 65 years (N = 1,843). Coagulation parameters were assessed in a subset of randomized subjects who received BZA 20 mg/CE 0.45 mg (n = 135), BZA 20 mg/CE 0.625 mg (n = 154), BZA 20 mg (n = 73), CE 0.45 mg/medroxyprogesterone acetate (MPA) 1.5 mg (n = 70), or PBO (n = 158).[br][bold]Results:[/bold] Coagulation parameters were evaluated in 590 dosed subjects (mean age [plusmn] standard deviation [SD], 52.9 years [plusmn] 3.4 years; mean years since last menstrual period [plusmn] SD, 2.5 years [plusmn] 1.5 years). At 12 months, BZA 20 mg/CE 0.45 and 0.625 mg and BZA 20 mg showed decreases from baseline in antithrombin III while PBO showed an increase (mean percent change of [ndash]2.3%, [ndash]4.7%, [ndash]3.4%, and 3.7% for BZA 20 mg/CE 0.45 and 0.625 mg, BZA 20 mg, and PBO, respectively; [italic]P[/italic] [lt]0.0001 for BZA/CE and BZA alone vs PBO). The change in antithrombin III for CE 0.45 mg/MPA 1.5 mg (1.8%) was similar to that for PBO. There were no differences among the BZA 20-mg/CE 0.45- and 0.625-mg, BZA 20-mg, CE 0.45-mg/MPA 1.5-mg, and PBO groups in the mean percent changes in Protein C activity (19.3%, 10.8%, [ndash]5.1%, 5.2%, and 1.5%, respectively) or Protein S activity ([ndash]9.3%, [ndash]8.9%, [ndash]5.4%, 5.7%, and [ndash]4.5%, respectively). Compared with PBO, both BZA/CE doses showed decreases in fibrinogen and PAI-1 activity, and an increase in plasminogen ([italic]P[/italic] [lt]0.05 for all). Changes in D-dimer, PAI-1 antigen, prothrombin time, and partial thromboplastin time were generally similar among groups.[br][bold]Conclusion:[/bold] Overall, there was no evidence of clinically meaningful changes in coagulation parameters for BZA 20 mg/CE 0.45 and 0.625 mg compared with PBO at 12 months. Together with the efficacy of BZA/CE on menopausal symptoms and bone preservation, these findings indicate that BZA/CE may be a promising option for postmenopausal women.[br][br]Sources of Research Support: This study was supported by Pfizer Inc.[br][br]Disclosures: SM: Employee, Pfizer, Inc. KAR: Employee, Pfizer, Inc. JRT: Employee, Pfizer, Inc. AAC: Employee, Pfizer, Inc. 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2123 49 150 SAT-29 PO32-01 Saturday 89 2012


90 ENDO12L_SAT-30 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) A Pooled Analysis of the Effects of Bazedoxifene/Conjugated Estrogens on Lipid Parameters from the Selective Estrogens, Menopause, and Response to Therapy Trials Sebastian Mirkin, Kelly A Ryan, Kaijie Pan, Arkadi A Chines, Rogerio A Lobo Pfizer Inc, Collegeville, PA; Columbia University Medical Center, New York, NY [bold]Objective:[/bold] Bazedoxifene (BZA)/conjugated estrogens (CE) is the first tissue selective estrogen complex in clinical development. The Selective estrogens, Menopause, And Response to Therapy (SMART) trials showed that BZA/CE reduced menopausal symptoms and bone loss and was well tolerated in postmenopausal women with a uterus. The effects of BZA/CE on lipid parameters were assessed based on pooled data from the SMART-1, SMART-2, SMART-3, SMART-4, and SMART-5 trials.[br][bold]Methods:[/bold] The SMART trials were randomized, double-blind, placebo (PBO)-controlled, phase 3 studies in non-hysterectomized postmenopausal women. Lipid parameters were assessed at 12 and 24 months for the SMART-1 trial, 3 months for the SMART-2 and SMART-3 trials, and 12 months for the SMART-4 and SMART-5 trials; pooled data were analyzed for women treated with BZA 20 mg/CE 0.45 or 0.625 mg or PBO (N = 4,735).[br][bold]Results:[/bold] BZA 20 mg/CE 0.45 and 0.625 mg were associated with decreases from baseline in total cholesterol (mean changes [95% confidence intervals (CI)] of [ndash]0.3 [[ndash]1.7, 1.1] and [ndash]0.3 [[ndash]1.6, 1.0] mmol/L, respectively, at 12 months; [ndash]0.1 [[ndash]1.6, 1.3] mmol/L for PBO). BZA 20 mg/CE 0.45 and 0.625 mg also reduced levels of low-density lipoprotein cholesterol (LDL-C) from baseline (mean changes [95% CI] of [ndash]0.4 [[ndash]1.6, 0.9] and [ndash]0.4 [[ndash]1.5, 0.7] mmol/L, respectively, at 12 months; [ndash]0.1 [[ndash]1.3, 1.1] mmol/L for PBO). Similar results for total cholesterol and LDL-C were seen at 3 and 24 months. Changes in high-density lipoprotein cholesterol were similar among groups at all time points. The BZA/CE doses showed slight increases in triglycerides at 3, 12, and 24 months. Mean changes (95% CI) in triglycerides for BZA 20 mg/CE 0.45 and 0.625 mg and PBO at 12 months were 0.1 ([ndash]0.9, 1.2), 0.1 ([ndash]0.9, 1.2), and [ndash]0.01 ([ndash]1.0, 1.0) mmol/L, respectively. At 3, 12, and 24 months, the BZA/CE doses showed slight increases in apolipoprotein A (0.13-0.14, 0.16-0.18, and 0.14-0.15 g/L, respectively) compared with minimal changes for PBO, and slight decreases in apolipoprotein B ([ndash]0.05 to [ndash]0.07, [ndash]0.02 to [ndash]0.03, and [ndash]0.02 to [ndash]0.03 g/L, respectively) compared with an increase of 0.03 g/L for PBO at 12 and 24 months.[br][bold]Conclusion:[/bold] Based on this pooled analysis of the SMART studies, BZA 20 mg/CE 0.45 and 0.625 mg were associated with a favorable overall lipid profile in postmenopausal women over 2 years of treatment.[br][br]Sources of Research Support: This study was supported by Pfizer Inc.[br][br]Disclosures: SM: Employee, Pfizer, Inc. KAR: Employee, Pfizer, Inc. KP: Employee, Pfizer, Inc. AAC: Employee, Pfizer, Inc. Nothing to Disclose: RAL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2170 49 151 SAT-30 PO32-01 Saturday 90 2012


91 ENDO12L_SAT-31 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Low Risk of Thrombophilia with Bazedoxifene/Conjugated Estrogens: Summary of Pooled Coagulation Data from the Selective Estrogens, Menopause, and Response to Therapy Trials Sven O Skouby, Rogerio A Lobo, Kelly A Ryan, John R Thompson, Barry S Komm, Arkadi A Chines, Sebastian Mirkin Herlev Hospital, Herlev, Denmark; University of Copenhagen, Copenhagen, Denmark; University of Southern Denmark, Esbjerg, Denmark; Columbia University Medical Center, New York, NY; Pfizer Inc, Collegeville, PA [bold]Objective:[/bold] Bazedoxifene (BZA)/conjugated estrogens (CE) is a menopausal therapy in development and has shown efficacy in reducing menopausal symptoms and bone loss in the Selective estrogens, Menopause, And Response to Therapy (SMART) trials in postmenopausal women. The effects of BZA/CE on coagulation parameters were assessed based on pooled data from the SMART-1, SMART-4, and SMART-5 trials.[br][bold]Methods:[/bold] The SMART-1, SMART-4, and SMART-5 trials were randomized, double-blind, placebo (PBO)- and active-controlled, phase 3 studies in postmenopausal women with a uterus. Mean changes in coagulation parameters were assessed at 12 months in the SMART-4 and SMART-5 trials and at 12 and 24 months in the SMART-1 trial; pooled data were analyzed for women who received BZA 20 mg/CE 0.45 or 0.625 mg or PBO.[br][bold]Results:[/bold] Across the 3 trials, coagulation parameters were assessed in 1,978 randomized and dosed subjects. At 12 months, BZA 20 mg/CE 0.45 and 0.625 mg showed decreases from baseline in antithrombin III compared with an increase for PBO ([ndash]0.05, [ndash]0.06 and 0.02 L/L, respectively; [italic]P[/italic] [lt]0.01 vs baseline for all). At 24 months, BZA 20 mg/CE 0.45 and 0.625 mg and PBO showed similar decreases in antithrombin III ([ndash]0.27, [ndash]0.26, and [ndash]0.21 L/L, respectively; [italic]P[/italic] [lt]0.001 vs baseline for all). Changes in Protein C activity at 12 and 24 months were [ndash]0.01 to [ndash]0.03 and 0.05 L/L, respectively, for the BZA/CE doses compared with [ndash]0.02 and 0.04 L/L, respectively, for PBO ([italic]P[/italic] [lt]0.01 vs baseline for all except BZA 20 mg/CE 0.625 mg and PBO at 12 months). For the 2 BZA/CE doses and PBO, changes in Protein S activity were [ndash]0.02 to [ndash]0.03 and [ndash]0.02 L/L, respectively, at 12 months and 0.06 to 0.08 and 0.15 L/L, respectively, at 24 months ([italic]P[/italic] [lt]0.05 vs baseline for all except PBO at 12 months). There were no apparent changes in fibrinogen, prothrombin time, partial thromboplastin time, PAI-1 activity, plasminogen, and D-dimer for BZA/CE compared with PBO. Pooled incidences of venous thromboembolic and cerebrovascular events were low ([le]0.2%) and similar among groups.[br][bold]Conclusion:[/bold] BZA 20 mg/CE 0.45 and 0.625 mg showed an overall favorable coagulation profile in postmenopausal women, as changes in coagulation parameters were generally comparable to those for PBO over 2 years. Combined with the demonstrated efficacy and safety of BZA/CE in treating menopausal symptoms and preventing osteoporosis, these findings show that BZA/CE may be a promising alternative for non-hysterectomized women.[br][br]Sources of Research Support: This study was supported by Pfizer Inc.[br][br]Disclosures: SOS: Advisory Group Member, Wyeth Pharmaceuticals. KAR: Employee, Pfizer, Inc. JRT: Employee, Pfizer, Inc. BSK: Employee, Pfizer, Inc. AAC: Employee, Pfizer, Inc. SM: Employee, Pfizer, Inc. Nothing to Disclose: RAL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2187 49 152 SAT-31 PO32-01 Saturday 91 2012


92 ENDO12L_SAT-32 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Vasomotor Symptoms and Cardiovascular Risk Markers in Postmenopausal Women Emily D Szmuilowicz, JoAnn E Manson, Ellen W Seely Northwestern University, Chicago, IL; Brigham and Women[apos]s Hospital and Harvard Medical School, Boston, MA; Brigham and Women[apos]s Hospital, Boston, MA [underline]Background[/underline]: We previously found that women who had vasomotor symptoms (VMS) around the time of menopause had lower risk of CVD, stroke, and all-cause mortality and that the predictive value of VMS for clinical CVD events varied by the timing of VMS onset.[br][underline]Methods[/underline]: We examined associations between VMS and both circulating CVD risk biomarkers and blood pressure (BP) in the Women[apos]s Health Initiative Observational Study (WHI-OS). N ranged 649-662 for lipid measures, 2327-2360 for biomarkers, and was [gt]59000 for BP and white blood cell count (WBC) measurements. VMS status was defined as: (1) No VMS: no VMS at menopause onset or at WHI-OS enrollment; (2) Early VMS: VMS at menopause onset, but not at WHI-OS enrollment; (3) Persistent VMS: VMS at both menopause onset and WHI-OS enrollment; (4) Late VMS: VMS at WHI-OS enrollment, but not at menopause onset. The associations between VMS and outcome variables were examined using linear regression. Outcome variables were systolic BP (SBP), diastolic BP (DBP), C-reactive protein (CRP), interleukin-6 (IL-6), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), E-selectin, white blood cell count (WBC), insulin, glucose, and lipid levels. Covariates included: age, ethnicity, education, smoking, hysterectomy/oophorectomy status, aspirin use, hormone therapy use, and physical activity (partial adjustment); and above plus BMI, hypercholesterolemia, diabetes, and family history of CHD (full adjustment).[br][underline]Results[/underline]: Compared to no VMS, persistent VMS were associated with higher SBP and DBP, and late VMS were associated with higher SBP (but not DBP) in the partially-adjusted model only. Early VMS were not associated with BP in either model. Persistent and late (but not early) VMS were associated with higher WBC in the partially-adjusted model only. In both models, persistent (but not early/late) VMS were associated with higher glucose and insulin, and early (but not persistent/late) VMS were associated with lower VCAM-1. There were no consistent associations between VMS and CRP, IL-6, ICAM-1, E-selectin, or lipids.[br][underline]Conclusions[/underline]: Persistent and late VMS were associated with one or more CVD risk markers (higher BP, WBC, and/or markers of impaired glucose metabolism), while early VMS were associated with lower levels of one marker of endothelial dysfunction. The pathways linking VMS to CVD risk may be mediated by traditional CVD risk factors and biomarkers and may be influenced by the timing of VMS.[br][br]Nothing to Disclose: EDS, JEM, EWS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2236 49 153 SAT-32 PO32-01 Saturday 92 2012


93 ENDO12L_SAT-33 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Cyclic Pattern of 17 [beta]-Estradiol Improves Spatial Memory Performance and Ischemia Outcome in Female Rats Ami Raval, Raquel Borges-Garcia, W Javier Moreno, Isabel Saul, Helen Bramlett Univeristy of Miami, Miami, FL; Univeristy of Miami, Miami, FL; University of Miami, Miami, FL The failure of the Women[apos]s Estrogen for Stroke Trial raised concern regarding the safety of chronic estrogen treatment in women. Recently, we demonstrated that an exogenous bolus of 17[beta]-estradiol to ovariectomized rats protect hippocampus against ischemia via activation of cyclic-AMP response element binding protein (CREB) pathway. In brain phosphorylation of CREB regulates learning and memory. Therefore, in the current study we hypothesize that intermittent estrogen treatment improves cognition and protects the brain from ischemic damage in female rats. Seven days after ovariectomy female rats were injected with 17[beta]-estradiol (5 [micro]g/Kg; i.p.) or oil at intervals of every 48/72h for the period of one month. During last 7 days rats (n=11) were tested using the Morris watermaze to measure cognitive capabilities. The first 4 days measured learning capabilities (learning the position of a platform), the 5[sup]th[/sup] day measured memory (memory of platform location when platform is removed), and the 6[sup]th[/sup] and 7[sup]th[/sup] days measured working memory (learning new platform location). In parallel set of experiments, on 48 hours after last hormone treatment rats were exposed to cerebral ischemia produced by 10 min of bilateral carotid occlusion and systemic hypotension (50 mmHg). Seven days later, rat brains were fixed for histopathological assessment. Results the watermaze study demonstrated that there was a difference in performance between the estradiol and vehicle groups. Data is presented as mean and standard deviation. Experimental group required less time to find the platform (21[plusmn]11s), than the control group (24[plusmn]18s), and remained in the correct quadrant longer during their allotted time (52%vs.12%). Secondly, in the working memory trials, treated rats also required less time to find the platform (21[plusmn]10s) than the control group (25[plusmn]11s). Further results are forthcoming. The results of histopathological analysis demonstrated that the number of live neurons per slice in the CA1 hippocampal region in na[iuml]ve rats was 1100[plusmn]45 (n=4). Ischemia in OvX rats decreased the number of normal neurons by 82% (192[plusmn]10, n= 6, p[lt]0.05). Intermittent estradiol-17[beta] treatment of OvX rats prior to ischemia increased the number of normal neurons 51% (556[plusmn]13, n=8, every 48h) and 41% (446[plusmn]14, n=7, every 72h) compared to the OvX group (p[lt]0.05). Intermittent 17[beta]-estradiol treatment conferred protection in cognitive tasks against ischemia in OvX rats.[br][br]Nothing to Disclose: AR, RB-G, WJM, IS, HB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2270 49 154 SAT-33 PO32-01 Saturday 93 2012


94 ENDO12L_SAT-34 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Effect of Oral Contraceptive Use on Heart Rate Variability during Laboratory-Induced Stress Lacey Wisslead, Lauren Drogos, Leah Rubin, Antonia Savarese, Erin Eatough, Kristen Mordecai, Pauline Maki University of Illinois at Chicago, Chicago, IL; University of Illinois at Chicago, Chicago, IL; University of South Florida, Tampa, FL; Baltimore VA Medical Center, Baltimore, MD Background: Endogenous estrogen plays a critical role in the regulation of autonomic function, including cardiac control as measured by heart rate variability (HRV). Changes in HRV have been observed across the rat estrous cycle. In addition, blockade or removal of endogenous hormones eliminates the corresponding change in HRV (1). Clinical studies have demonstrated that estrogen therapy restores autonomic balance in postmenopausal women, (2) and estrogen, but not progesterone correlates with HRV in healthy young women (3). HRV in young women using oral contraceptives has not been sufficiently studied. The only study to date suggested that OCs do not impact autonomic tone; however, it is unknown if HRV changes between active and inactive pill phases in oral contraceptive users. HRV is also affected by stress (4). In the present study, we examined the effects of progestin-containing OCs on HRV, specifically respiratory sinus arrhythmia (RSA), during conditions of stress and non-stress. We hypothesized that effects of oral contraceptives on RSA will become more pronounced during conditions of stress.[br]Methods: Participants included healthy women aged 18-40 (n = 30; 14 active; 16 inactive pill phase) who were taking the same estrogen-based oral contraceptive for a minimum of six months prior to study entry. Each participant contributed HRV measures obtained under two experimental conditions, one a laboratory-induced stress condition and the other a parallel control condition. HRV data were collected throughout the experimental and control sessions via a UFI Biolog monitor. HRV data were separated into 10 segments per experimental session, and each segment was edited using Cardio Edit.[br]Results: All women showed an increase in stress and anxiety following the TSST (p[apos]s[lt].001).There was a trend for RSA to be lower during the stress compared to the non-stress condition, F(1,28) =3.13, p=0.08, eta2=0.10. Unexpectedly, there were no significant differences between oral contraceptive users in the active versus inactive pill phase on RSA during the stress or non-stress condition, F(1,28) =1.66, p=0.21, eta2=0.06.[br]Conclusions: Contrary to predictions, oral contraceptive use (being in the active pill phase) did not differentially impact HRV during stress. Consistent with previous work in healthy women, our results suggest that there were no significant differences demonstrated on cardiac neural regulation.[br][br](1) T. B. Kuo, C. T. Lai, F. C. Hsu et al., Endocrinology 15 (6), 2613. (2) I. Virtanen, O. Polo, P. Polo-Kantola et al.,. Maturitas 37 (1), 45 (2000). (3) A. S. Leicht, D. A. Hirning, and G. D. Allen, Experimental physiology 88 (3), 441. (2003). (4) A. V. Klinkenberg, U. M. Nater, A. Nierop et al., Acta. obstetricia et gynecologica Scandinavica 88 (1), 77 (2009).[br][br]Nothing to Disclose: LW, LD, LR, AS, EE, KM, PM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2333 49 155 SAT-34 PO32-01 Saturday 94 2012


95 ENDO12L_SAT-35 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Association between Copy Number Variation (CNV) Polymorphism in [italic]CNTNAP4[/italic] Gene and Aging in Women: Effect Modification by Estrogen Receptor Genetic Variant Leonid Iakoubov, Malgorzata Mossakowska, Magdalena Owczarz, Monika Puzianowska-Kuznicka GeneCona, LLC, Vallejo, CA; International Institute of Molecular and Cell Biology, Warsaw, Poland; Medical Center of Postgraduate Education, Warsaw, Poland; Mossakowski Medical Research Centre, Warsaw, Poland [bold]BACKGROUND:[/bold] Accumulation of myelin-related abnormalities is characteristic for some neuropsychiatric disorders, as well as for normal aging. Several genes from neurexin superfamily encode proteins that critically support the integrity of myelin-covered fibers. Polymorphisms in a few of these genes are associated with autism, epilepsy, and schizophrenia. At least some of associations are sex-specific. We investigated whether CNV polymorphisms in neurexin superfamily genes are associated with aging in a similar sex-related fashion, and if female-specific associations could be affected by a functional polymorphism in the estrogen receptor gene [italic]ESR1[/italic].[br][bold]METHODS:[/bold] Five common intronic CNVs of in-del type that belong to 4 genes from neurexin superfamily were genotyped using TaqMan qPCR. To evaluate the association with aging, polymorphism frequency was compared between two groups of community old adults aged 65-75 and 80-90. The conclusions are based on data from two studies, a discovery study in USA Caucasians (n=90), and replication study in Eastern European Caucasians (n=1296). In each of them, samples from age groups, as well as male/female-originating groups were equally represented. Statistical significance was evaluated using Fisher[apos]s 2x2 exact test. Due to restricted power of USA sample-based study, effects of [italic]ESR1[/italic] variants on the[italic]CNTNAP4[/italic] gene association with aging were tested in Europe sample-based study only and need replication.[br][bold]RESULTS:[/bold] The only association with aging that was statistically significant in each of the two studies was for the CNV in [italic]CNTNAP4[/italic] gene in females. Combined, there were 48% carriers of risk polymorphism in females aged 65-75, versus 36% in aged 80-90, OR=1.65; CI(1.21-2.23) Fisher[apos]s P=0.00127. After additional stratification by [italic]ESR1[/italic] CNV genotype, the difference remained statistically significant in carriers of [italic]ESR1[/italic] non-deletion variant (OR =1.76; CI 1.19-2.59; P=0.00466), while virtually absent from carriers of the deletion allele (OR=0.93; CI 0.53-1.61; P=0.88829).[br][bold]CONCLUSION:[/bold] CNV in [bold][italic]CNTNAP4[/italic][/bold] gene is associated with aging in females. The association is not observed in a subpopulation of female carriers of a functional polymorphism in the [bold][italic]ESR1[/italic][/bold] gene that encodes estrogen receptor with higher sensitivity to the ligand. Female stratification based on variations in [bold][italic]CNTNAP4[/italic][/bold] and [bold][italic]ESR1[/italic][/bold] genes might reveal subpopulations with differential aging consequences in response to modifications in sex hormone status.[br][br]Nothing to Disclose: LI, MM, MO, MP-K 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 576 49 156 SAT-35 PO32-01 Saturday 95 2012


96 ENDO12L_SAT-36 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Simultaneous Measurement of Fourteen Steroid Hormones across the Menstrual Cycle in Healthy Women Using Liquid Chromatography-Tandem Mass Spectrometry Candace Keefe, Richard E Reitz, Corrine Welt, Mildred Goldman, Nigel Clarke Massachusetts General Hospital, Boston, MA; Quest Diagnostics Nichols Institute, San Juan Capistrano, CA Context: Steroid immunoassays and radioimmunoassays have limited sensitivity, specificity and accuracy. These limitations can make it difficult to assess hormone levels in children, women and hypogonadal adults. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) can provide improved accuracy and sensitivity. We have now validated a LC-MS/MS that simultaneously measures 14 steroid hormones, with results reported in reproductive age women.[br]Methods: Daily blood samples were obtained from healthy women with regular, 25-35 day menstrual cycles (n=35). Menstrual cycles were standardized to 28 days and hormone levels were centered to ovulation for comparison across the follicular and luteal phases. Pelvic ultrasounds were performed to ensure the emergence of a dominant follicle. Hormone levels were compared in the early, mid and late follicular phase (EFP, MFP, LFP) and luteal phase (ELP, MLP, LLP) and at the midcycle gonadotropin surge (MGS).[br]Results: Progesterone, 17-hydroxyprogesterone, testosterone and androstenedione levels varied across the cycle. In the follicular phase, 17OH progesterone levels increased at the MGS compared to the EFP [177 (67, 349) vs. 52 (20, 93) ng/dL; median (2.5, 97.5 percentile); p[lt]0.001] and preceded the rise in progesterone. In the MLP, 17OH progesterone [261 (139, 431) vs. (59 (23, 102) ng/dL; MLP vs. MFP; p[lt]0.001] and progesterone levels [15 (6.7, 22.2) vs. (0.1 (0.1, 0.3) ng/mL; p[lt]0.001] were higher than in the MFP, and the LP pattern of 17OH progesterone mirrored that of progesterone. Testosterone [42 (24, 66) vs. 26 (16, 47), 30 (20, 54) ng/dL; MGS vs. EFP, LLP; p[lt]0.001] and androstenedione levels [136 (73, 230) vs. 88 (58, 182), 98 (68, 175) ng/dL; p[lt]0.001] were highest at midcycle. Cortisone was lower in the LP than in the FP (p=0.02), but cortisol and DHEA levels did not change across the cycle.[br]Conclusion: This study validates previous findings that testosterone and androstenedione levels peak at the midcycle and 17-hydroxyprogesterone levels mirror the rise and fall of progesterone in the luteal phase. The assay will be a powerful tool for examining multiple adrenal and gonadal steroids simultaneously, which will be useful in the diagnosis of congenital adrenal hyperplasias.[br][br]Disclosures: RER: Chairman, Quest Diagnostics. MG: Employee, Quest Diagnostics. NC: Employee, Quest Diagnostics. Nothing to Disclose: CK, CW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2363 49 157 SAT-36 PO32-01 Saturday 96 2012


97 ENDO12L_SAT-37 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Analysis of Estrogens in Plasma Samples by LC-MS/MS, with a Reduced Sample Preparation Using SelexION Ion Mobility Technology To Remove Interferences Stefanie Krepenhofer, Bruno Casetta, Michael Jarvis, Michal Weinstock AB SCIEX, Concord, Canada (For Research Use Only. Not for Use in Diagnostic Procedures) The two estrogens, Estrone (E1) and 17[beta]-Estradiol (E2) are the primary female sex hormones. The objective of this work was to develop an accurate LC-MS/MS analytical method to detect Estrone (E1) and 17[beta]-Estradiol (E2), using simplified sample preparation to save time and cost. In general, the analysis of these compounds by LC-MS/MS is challenging due to the large number of interferences in addition to high chemical noise observed in the data. Therefore, we have evaluated the use of differential mobility spectrometry (DMS) to improve the selectivity of the detection, which has enabled us to achieve lower Limits of Detection (LOD) for the estrogens in plasma samples, after a simple sample preparation consisting only of protein precipitation.[br]Protein was precipitated from anonymized plasma samples using acetonitrile. After centrifugation the supernatant was directly injected on a 2D chromatographic separation system using a C8 clean-up column (10 [mu]m, 50 x 4.6 mm) and a Phenomenex Luna C8 separation column (5 [mu]m, 100 x 2.1 mm). The HPLC separation used a 10 minute flow gradient of methanol and water containing 0.2 mM NH[sub]4[/sub]F. The mass spectrometric detection of the estrogens was accomplished using an ABSCIEX QTRAP[reg] 5500 system operating in positive ionization mode. For improved selectivity and sensitivity, the instrument was equipped with the SelexION[trade] differential mobility device, which enabled the removal of potential interferences based on differences in their ion mobility characteristics. As a result of the additional selectivity provided by the SelexION device, a significant reduction of the background interferences was achieved. Using this approach it was possible to detect an Estrone (E1) concentration below 1 ppt and an 17[beta]-Estradiol (E2) concentration below 5 ppt in plasma, using the simple sample preparation described.[br][br]Disclosures: SK: Employee, AB Sciex. BC: Coinvestigator, AB Sciex. MJ: Employee, AB Sciex. MW: Employee, AB Sciex. 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2231 49 158 SAT-37 PO32-01 Saturday 97 2012


98 ENDO12L_SAT-38 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Estradiol Measurement Inter-Laboratory Comparison Study and Commutability Assessment Julianne Cook Botelho, Megan Vidal, Matthew Gatcombe, Hans Cooper, Hubert W Vesper Centers for Disease Control and Prevention, Atlanta, GA Accurate laboratory results are a matter of general concern in clinical medicine. This also holds true for results from Estradiol (E2) testing, especially at lower levels found in postmenopausal women. The goal of the CDC Hormone Standardization Program is to improve diagnosis, treatment, prevention and research of diseases and disorders by standardization of hormone measurements. This Inter-Laboratory Study is the first step in this process for E2 measurements.[br]An assessment of E2 measurement has been made by 17 laboratories with 23 different platforms from clinical, commercial, and research laboratories which include mass spectrometry based laboratory developed tests (LDTs) as well as immunoassay manufacturers and end users. Participants measured 40 single donor adult male and female (pre and postmenopausal) samples in duplicate over 3 different days over a range of 2.5-300 pg/mL. The serum material used for this study was produced following a standard protocol developed by the Clinical Laboratory and Standards Institute (CLSI Protocol C37-A) shown to contain minimal matrix effects and to behave similar to regular patient samples. The second part of this study was to assess the commutability of E2 standard reference materials (SRM 971) at 2 levels and of 9 commercial testing materials (CTM) used in external quality assurance (EQA) programs or as potential calibrator materials. Knowledge about the commutability of a material is important to assess its suitability as calibrators or trueness controls. The commutability assessment will evaluate whether SRMs and CTMs behave similarly as patient samples on an analytical system.[br]Current suggested criteria based on biological variability for estradiol recommend an acceptable bias of [plusmn]8.3%, imprecision of [le]11.4%, and a total error limit of [plusmn]27.2%. Bias assessments were made according to CLSI Protocol EP9-A2 and commutability by CLSI Protocol C53-A. These assessments will provide information about the measurement performance of individual assays, the measurement variability across laboratories, the bias of methods as compared to a reference method and an assessment of material suitability for calibrators or trueness controls. Furthermore, this study will help in better defining current needs and gaps in E2 measurements, which will help with standardizing the measurement more efficiently.[br][br]Nothing to Disclose: JCB, MV, MG, HC, HWV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2237 49 159 SAT-38 PO32-01 Saturday 98 2012


99 ENDO12L_SAT-39 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Testosterone Measurement by Mass Spectrometry [mdash] A Tale of Three Internal Standards Laura Jane Owen, Brian George Keevil UHSM, Manchester, UK Introduction[br]Testosterone measurement by liquid chromatography tandem mass spectrometry (LC-MS/MS) is well accepted as the preferred technique for the analysis of serum testosterone in both males and particularly females. However variation is seen between LC-MS/MS assays and is most likely to be due to method differences between laboratories. One area of inconsistency amongst routine LC-MS/MS assays is the choice of internal standard. We investigated the effects of three internal standards on the results obtained.[br]Methods[br]Stable isotopes of testosterone were prepared in methanol. Testosterone with two deuterium (D2), five deuterium (D5) and three carbon thirteen enrichment (C13) were separately assessed.[br]Sera, calibrators and quality controls (100 [micro]L) were extracted using 0.5 mL of ether following the addition of 10 [micro]L of internal standard. All aliquots were prepared in triplicate, one for each type of internal standard. After mixing the ether was transferred to the wells of a 96-deep well block then evaporated to dryness. Extracts were reconstituted with 100 [micro]L of 50% mobile phases and analysed using a Waters Acquity LC and Quattro Premier tandem mass spectrometer. This method had previously shown to have excellent agreement with a reference method using the D2 internal standard.[br]Results[br]Lower results were obtained when using D5 testosterone when compared to D2 testosterone. The Passing-Bablock regression equation was; testosterone (D5) nmol/L = 0.86 x testosterone (D2) + 0.04.[br]The C13 internal standard also gave lower results but was closer than the D2 target than the D5 internal standard. The Passing-Bablock regression analysis was; testosterone (C13) nmol/L = 0.90 x testosterone (D2) + 0.02.[br]Discussion[br]The choice of internal standard alone can have a significant affect on the results obtained by LC-MS/MS assays for testosterone using this chromatography. The effects of the combination of chromatography and internal standard choice should be investigated during method development.[br][br]Nothing to Disclose: LJO, BGK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 553 49 160 SAT-39 PO32-01 Saturday 99 2012


100 ENDO12L_SAT-40 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Dynamics of Serum Testosterone during the Menstrual Cycle Evaluated by Daily Measurements with an ID-LC-MS/MS Method and a 2[sup]nd[/sup] Generation Automated Immunoassay Hong N Bui, Patrick M Sluss, Stuart Blincko, Marinus A Blankenstein, Annemieke C Heijboer VU University Medical Center, Amsterdam, Netherlands; Massachusetts General Hospital, Boston, MA; Abbott, Wiesbaden, Germany [bold]Context: [/bold]Testosterone levels in normally cycling women are generally assumed to be elevated around the time of ovulation. The clinical relevance of changing testosterone levels during the menstrual cycle, however, is unclear. Poor performance of current direct immunoassays for testosterone at low concentrations confounds this issue.[br][bold]Objective[/bold]: To assess daily testosterone fluctuation during the menstrual cycle by a thoroughly validated isotope dilution-liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) method and to evaluate whether a ARCHITECT[sup][reg][/sup] 2nd Generation Testosterone fully automated direct immunoassay is equally suited for this purpose.[br][bold]Design[/bold][bold]:[/bold] Serum samples obtained daily during the menstrual cycles of 25 healthy women, characterized by biochemical and physical examination.[br][bold]Measurements: [/bold]Testosterone, progesterone, estradiol, LH, FSH, and SHBG were measured.[br][bold]Results:[/bold] Performance of the ID-LC-MS/MS method (1) was concordant with a published reference method (2)(y = 1.007 x [ndash] 1.61 ng/dL; r = 0.9998). Comparison of the ARCHITECT[sup][reg][/sup] 2nd Generation Testosterone results to ID-LC-MS/MS yielded y = 1.095 x + 3.00 ng/dL (r = 0.9031). Overall, testosterone levels were significantly higher mid-cycle, but a peak was not discernable in each individual. Intra-individual variation exceeded the group average in the menstrual cycle; the ratio of extremes (max/min) found in individuals ranged from 1.6-3.7. Apart from a persistent positive bias, the immunoassay measured the same testosterone profiles as the ID-LC-MS/MS method. The reference interval was 8.65-48.7 ng/dL (0.30-1.69 nmol/L) for ID-LC-MS/MS and 14.4-57.6 ng/dL (0.50-2.00 nmol/L) for the 2[sup]nd[/sup] generation Architect.[br][bold]Conclusion: [/bold]Our ID-LC-MS/MS method measured low testosterone levels accurately. The ARCHITECT[sup][reg][/sup] 2nd Generation Testosterone immunoassay had acceptable performance across the entire range measured in women. On average, the elevation of mid-cycle testosterone levels is statistically significant, although not relevant for clinical practice since the day-to-day variation is higher and independent of the menstrual cycle. In this light, we recommend measuring testosterone on at least 2 independent occasions for diagnostic purposes.[br][br](1) Bui HN et al., Ann Clin Biochem 2010; 47: 248-252. (2) Thienpont LM et al., Clin Chem 2008; 54: 1290-1297.[br][br]Disclosures: SB: Employee, Abbott Laboratories. Nothing to Disclose: HNB, PMS, MAB, ACH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 611 49 161 SAT-40 PO32-01 Saturday 100 2012


101 ENDO12L_SAT-41 POSTER SESSION: Female Reproductive Endocrinology: Pregnancy [amp] Menopause (1:30 PM-3:30 PM) Reference Ranges for Circulating Testosterone in the Follicular and Luteal Phases of the Menstrual Cycle in a Healthy, Community-Based Sample of Women in the Framingham Heart Study Andrea D Coviello, Kerrie P Nelson, Shalender Bhasin Boston University School of Medicine, Boston, MA; Boston University School of Public Health, Boston, MA [bold]Background:[/bold] Normative ranges of circulating total (TT) and free testosterone (FT) in healthy women across the menstrual phase have not been well characterized; in part due to lack of sufficiently sensitive and reliable testosterone assays that allow for accurate measurement of testosterone in the low concentrations found in women.[br][bold]Objective:[/bold] To formulate reference ranges for TT and FT in healthy women with regular cycles from a community based population using liquid chromatography tandem mass spectroscopy (LC-MSMS).[br][bold]Methods:[/bold] TT and SHBG were measured in women from Generation 3 of the Framingham Heart Study (FHS). Women, 19-45 years, were included if they reported regular menstrual cycles between 28-35 days and were not taking hormonal contraceptives. Women were excluded if they were under or overweight (BMI[lt]18 or [gt]25 kg/m[sup]2[/sup]), smoked, had impaired fasting glucose, diabetes, hypertension, hyperlipidemia, cardiovascular disease or cancer. Menstrual cycle length, first day of the last menstrual cycle, and the regularity of menstrual cycles was obtained during the FHS examination. Based on these responses and the date of blood draw, women were classified as being in the follicular (first 10 days), luteal (last 14 days), or ovulatory phase. TT was measured by LC-MSMS (sensitivity 2 ng/dL), SHBG by immunofluorometric assay, and FT was calculated. Hormones were log transformed for analyses. [bold]Results: [/bold]Mean[plusmn]SD age 37[plusmn]6 years, BMI 22.0[plusmn]1.6 kg/m[sup]2[/sup], fasting glucose 87[plusmn]5 mg/dl, total cholesterol 170[plusmn]25 mg/dl, HDLc 64[plusmn]15 mg/dl. Hormones are presented as the median (2.5[sup]th[/sup], 97.5[sup]th[/sup] percentile range). [underline]Follicular Phase (n=67):[/underline] TT 23.7 (8.3, 48.3) ng/dl, FT 1.8 (0.7, 4.4) pg/ml, and SHBG 89.7 (43.0, 184.4) nM. [underline]Ovulatory Phase (n=33):[/underline] TT 34.7 (10.1, 48.3) ng/dl, FT 2.8 (1.1, 4.6) pg/ml, and SHBG 93.3 (28.3, 154.3) nM. [underline]Luteal Phase (n=101):[/underline] TT 28.5 (11.3, 62.5) ng/dl, FT 2.4 (0.9, 7.2) pg/ml, SHBG 96.1 (42.9, 161.3) nM. Total and Free T varied across phase (TT p=0.001, FT p=0.002) but SHBG did not (p=0.7). Total and Free T were significantly lower in the follicular phase compared to the luteal phase (p[lt]0.01).[br][bold]Conclusion: [/bold]We present reference ranges for TT and FT measured by LC-MSMS in healthy premenopausal women with regular menstrual cycles. Both TT and FT were significantly different between the follicular and luteal phases suggesting that timing of measurement with the menstrual cycle is important when assessing premenopausal women[apos]s androgen status.[br][br]Sources of Research Support: RO1 HL094755.[br][br]Nothing to Disclose: ADC, KPN, SB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1684 49 162 SAT-41 PO32-01 Saturday 101 2012


102 ENDO12L_SAT-42 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Dexamethasone Decreases Expression of Genes Involved in Cholesterol Biosynthesis and Steroidogenesis in Stallion Testes Nancy H Ing, David W Forrest, Penny K Riggs, Adrianna L Lee, Shavahn Loux, Charles C Love, Dickson D Varner, Steve P Brinsko, Thomas H Welsh Texas A[amp]M University, College Station, TX; Texas A[amp]M University, College Station, TX Glucocorticoids negatively impact testicular steroidogenesis in males of every mammalian species studied but the molecular mechanisms that are responsible remain unclear. High levels of glucocorticoids may arise from stress or medical syndromes or treatments. In male horses (stallions) and men, subfertility is not uncommon. In three mature stallions, a single dose of dexamethasone (DEX, 0.1 mg/kg i.v.) decreased the peripheral blood concentrations of testosterone by 70% within 12 h, while four control stallions were unaffected. Castration at that time yielded testicular tissues for analyses of global gene expression using microarrays. Of thirteen differentially expressed genes identified, seven were confirmed on northern blots or by quantitative RT-PCR. Expression of one gene (NFkappaB inhibitor alpha or NFKBIA) increased by treatment, consistent with anti-inflammatory effects of DEX. Expression of six genes was depressed by DEX. The glucocorticoid receptor alpha (GR) gene, the protein product of which transduces DEX effects, was down-regulated by DEX as expected. The other five genes down-regulated were involved in cholesterol biosynthesis [squalene epoxidase and 24-dehydrocholesterol reductase] or steroidogenesis [luteinizing hormone receptor, glutathione S-transferase A3 (GSTA3) and P450 aromatase]. The GSTA3 protein is intriguing because it isomerizes the immediate precursors of testosterone and progesterone in non-rodent mammalian species, including man. We cloned the entire GSTA3 mRNA from stallion testes. The cDNA sequence localized the gene and regulatory sequences on horse chromosome 20. PCR primers were designed to selectively amplify GSTA3 and not other family members. The encoded GSTA3 protein contains the five amino acid residues that are critical for steroid isomerase activity. The protein will be expressed and analyzed for activity in the laboratory of Dr. Bengt Mannervik, an expert on the GSTA family of enzymes. Within the testes, Leydig cells express the GR gene most highly, as well as the genes depressed by DEX. Primary cultures of Leydig cells from stallion testes also showed dose-dependent regulation of GSTA3 and NFKBIA mRNA concentrations by DEX. Therefore, we propose a model in which DEX acts directly on Leydig cells to attenuate expression of a population of genes that results in decreased testosterone production. This mechanism may be useful therapeutically for altering steroidogenesis.[br][br]Sources of Research Support: American Quarter Horse Foundation grants to NHI and THW; Link Endowment.[br][br]Nothing to Disclose: NHI, DWF, PKR, ALL, SL, CCL, DDV, SPB, THW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1623 50 163 SAT-42 PO35-01 Saturday 102 2012


103 ENDO12L_SAT-43 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Mechanisms of WNT4 Action in Leydig Cell Function Pascal Bernard, Janelle Ryan, Dagmar Wilhelm, Vincent R Harley Prince Henry[apos]s Institute of Medical Research, Melbourne, Australia; The University of Queensland, Brisbane, Australia Male sexual differentiation requires the production of male-specific hormones from both fetal Sertoli and Leydig cells in the gonads. Fetal Leydig cells produce testosterone essential for the virilisation of the male embryo. Despite their crucial role, little is known about Leydig cell development and function. WNT4, a Wnt ligand, is important for both male and female gonadal development. In males, it regulates proper vascular development and androgen production. WNT4 regulates the expression of genes encoding enzymes in the testosterone biosynthetic pathway in mice and WNT4 heterozygous mutations in humans leads to excess production of androgens. While strong evidence suggests that WNT4 regulates testosterone synthesis at the transcriptional level, very little is known about its mechanism of action. A number of steroidogenic genes involved in the production of testosterone are transcriptionally regulated by nuclear hormone receptor Steroidogenic Factor 1 (SF1). One report indicates that WNT4 may regulates SF1-mediated transcriptional activation of steroidogenic genes, however the precise mechanism is unknown.[br]Using a specific antibody raised against Wnt4, we discovered that Wnt4 is specifically expressed in Leydig cells of the developing testis in mice. Using microarray expression analysis we have identified a new target gene of Wnt4, the prorenin receptor. The prorenin receptor is strongly expressed in Leydig cells and is absent in these cells in Wnt4 knockout mice. Since the prorenin receptor acts as a receptor for Wnt signalling, we hypothesize that Wnt4 upregulates the expression of the prorenin receptor and acts via the prorenin receptor to regulate steroidogenesis in the testis. We are investigating the role of Wnt4 and the prorenin receptor in mice using Wnt4 and prorenin receptor knockout models.[br][br]Nothing to Disclose: PB, JR, DW, VRH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1399 50 164 SAT-43 PO35-01 Saturday 103 2012


104 ENDO12L_SAT-44 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Hormone-Induced 14-3-3Gamma Adaptor Protein Regulates Steroidogenic Acute Regulatory Protein Activity and Steroid Biosynthesis in MA-10 Leydig Cells Yasaman Aghazadeh, Malena Rone, Josiph Blonder, Timothy Veenstra, Buck Hales, Martine Germaine Culty, Vassilios Papadopoulos McGill University, Montr[eacute]al, Canada; National Cancer Institute at Frederick, Frederick, MD; Southern Illinois University, Carbondale, IL; McGill University, Montr[eacute]al, Canada; McGill University, Montr[eacute]al, Canada Cholesterol is the sole precursor of steroid hormones in the body. The import of cholesterol to the inner mitochondrial membrane, the rate-limiting step in steroid biosynthesis, relies on the formation of a protein complex that assembles at the outer mitochondrial membrane called the transduceosome(1). The transduceosome contains several mitochondrial and cytosolic components, including the steroidogenic acute regulatory protein (STAR) (1). Human chorionic gonadotropin (hCG) induces [italic]d[/italic][italic]e novo[/italic] synthesis of STAR, a process shown to parallel maximal steroid production (2). In the hCG-dependent steroidogenic MA-10 mouse Leydig cell line, the 14-3-3Gamma protein was identified in native mitochondrial complexes by mass spectrometry and immunoblotting, and its levels increased in response to hCG treatment. The 14-3-3 proteins bind and regulate the activity of many proteins, acting via target protein activation, modification, and localization (3). In MA-10 cells, cAMP induces 14-3-3Gamma expression parallel to STAR expression. Silencing of 14-3-3Gamma expression potentiates hormone-induced steroidogenesis. Binding motifs of 14-3-3Gamma were identified in components of the transduceosome, including STAR. Immunoprecipitation studies demonstrate a hormone-dependent interaction between 14-3-3Gamma and STAR that coincides with reduced 14-3-3Gamma homodimerization. The binding site of 14-3-3Gamma on STAR was identified to be S194 in the STAR-related sterol-binding lipid transfer (START) domain, the site phosphorylated in response to hCG. Taken together, these results demonstrate that 14-3-3Gamma negatively regulates steroidogenesis by binding to S194 of STAR, thus keeping STAR in an unfolded state, unable to induce maximal steroidogenesis (4). Over time 14-3-3Gamma homodimerizes and dissociates from STAR allowing this protein to induce maximal mitoxhondrial steroid formation.[br][br](1)Papadopoulos V et al., Mol Cell Endocrinol 2007; 59-64. (2)Stocco DM et al.,Endocr Rev 1996; 221-44. (3)Aitken A, Plant Mol Biol 2002; 993-1010. (4)Arakane F et al.,J Biol Chem 1997; 32656-62.[br][br]Sources of Research Support: CHR MOP Grant 102647 awarded to VP. RI-MUHC fellowship awarded to YA.[br][br]Nothing to Disclose: YA, MR, JB, TV, BH, MGC, VP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 567 50 165 SAT-44 PO35-01 Saturday 104 2012


105 ENDO12L_SAT-45 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Developmental Changes of microRNA Profiles of Rat Leydig Cells: From Stem to Adult Ren-Shan Ge, Yufei Zhang, Kaiming Yuan, Zhijian Su, Ying Su, Xiaoheng Li, Qingquan Lian Wenzhou Medical College, Wenzhou, China; Mudanjiang Medical University, Mudanjiang, China; Jinan University, Guangzhou, China We have identified a developmental sequence that leads to the establishment of the adult Leydig cell (ALC). The first identified cell in the sequence is the progenitor Leydig cells. This fibroblast-like, luteinizing hormone receptor positive (LHR[sup]pos[/sup]) and 3[beta]-hydroxysteroid dehydrogenase positive (3[beta]HSD[sup]pos[/sup]) cell is shown to be from stem Leydig cell and then gives rise to the immature and adult Leydig cell. The hallmark of stem Leydig cell is its capability of self-renewal and of specifying into Leydig cell lineage. A fundamental unanswered question is how stem Leydig cells are specified, maintained and instructed to differentiate? The intrinsic cues including microRNAs (miRNAs) in stem Leydig cells possibly orchestrate the self-renewal and specification. In the present study, we systemically performed the miRNA arrays (653 common miRNAs) for stem, progenitor, immature and adult Leydig cells. We found that 3 miRNAs (miR-31, -221 [amp]-222) were 10 fold downregulation from stem to progenitor Leydig cells, with miR-31 downregulation by 600 fold. Using PicTar and TargetScanS, we find that Leydig cell specific transcription factor steroidogenic factor 1 3[apos] untranslated region (NR5A1-3[apos]UTR) has miR-31 binding site. miR-124 and 325 were significantly downregulated, mi-532-5p was upregulated from progenitor to immature Leydig cells. In conclusion, many miRNAs regulate the transition of stem Leydig cells in the Leydig cell lineage.[br][br]Sources of Research Support: The study is partially funded by NSFC 30871434.[br][br]Nothing to Disclose: R-SG, YZ, KY, ZS, YS, XL, QL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1417 50 166 SAT-45 PO35-01 Saturday 105 2012


106 ENDO12L_SAT-46 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Silver Nanoparticles Affect Leydig Cell Function [italic]In Vivo[/italic] Thomas Garcia, Guilherme Costa, Luiz Renato Franca, Marie-Claude Hofmann University of Illinois, Urbana, IL; Federal University of Minas Gerais, Belo Horizonte, Brazil Spermatogonial stem cells (SSCs) provide the foundation for spermatogenesis and are vital for the perpetuation of species. Previous studies from our lab examined the in vitro effects of silver nanoparticles (AgNPs) on an SSC cell line. Our data showed that AgNPs interfered with SSC proliferation in a dose and size-dependent manner and that AgNPs disrupted components of the GDNF signaling pathway, a pathway critical for self-renewal of SSCs in vivo. If AgNPs were to induce a similar effect in vivo, spermatogenesis and fertility would be severely impaired. To test this, CD1 male mice were given injections of vehicle and 1 mg/kg spherical AgNPs of 10nm diameter via the tail vein every 3 days, five times, beginning at postnatal day 45. This in vivo dose is roughly equivalent to the lowest in vitro dose with effects (10 [mu]g/ml) from our previous studies, and [sim]2,000 times greater than that found in the blood in a previous AgNP inhalation study. Although serum levels of LH and FSH were unaffected at 15, 60, and 120 days, serum and intratesticular levels of testosterone were significantly increased at 15 days. Additionally, significant changes in seminiferous epithelium morphology and Leydig cell nucleus and individual size (morphology) were observed at 15 and 60 days. Testis gene expression analysis of cholesterol biosynthesis, cholesterol transport, testosterone and estradiol biosynthesis, and GH-related genes revealed 3-[beta]-hydroxysteroid dehydrogenase mRNA significantly upregulated in treated animals at 15 days. In conclusion, we found significant histopathological alterations in Leydig cells, and significant changes in testicular testosterone levels and a critical biosynthesis gene. Thus, our results demonstrate that AgNPs have the potential to significantly impact testis health through altered steroid production.[br][br]Sources of Research Support: NIH T32 ES007326 and NIH HD054607.[br][br]Nothing to Disclose: TG, GC, LRF, M-CH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2328 50 167 SAT-46 PO35-01 Saturday 106 2012


107 ENDO12L_SAT-47 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Oxidative Stress Induced by the Xenoestrogen Bisphenol A in Rat Leydig Cells Is Influenced by Diet Manjunatha K Nanjappa, Manuj Ahuja, Muralikrishnan Dhanasekaran, Frank F Bartol, Robert L Judd, Benson T Akingbemi Auburn University, Auburn, AL; Auburn University, Auburn, AL Bisphenol A (BPA) is a high volume production chemical used in the manufacture of polycarbonate plastics and epoxy resins. Greater than 90% of the population in the United States has measurable BPA levels in their blood. Concern exists that exposure to BPA may predispose individuals to adverse health outcomes. Here, objectives were to determine if exposure to environmental chemical BPA and/or consumption of high energy diets induces oxidative stress in Leydig cells. Long Evans male rats (n=14) were exposed to BPA by maternal gavage at 0, 2.5 and 25 [micro]g/kg body weight from gestational day 12 to postnatal day (PND) 21 and were fed a BPA-free diet from weaning to PND 70. Subsequently, animals from each treatment group were maintained for a period of 28 days (PND 71 to 98) on a normal or high fat diet (NFD or HFD, Harlan-Teklad, Indianapolis, IN) with 16% or 42% of energy content from fat. Animals were sacrificed within 24 h of day 98 postpartum when serum and Leydig cells were obtained to assess oxidative stress and antioxidant enzyme activity. Serum levels of thiobarbituric acid reactive substance, a marker for oxidative stress, were greater in male rats exposed to BPA and maintained on HFD than in NFD control animals (P[lt]0.05). However, serum glutathione (GSH) levels increased only in animals exposed to the 25 [micro]g/kg dose of BPA and fed HFD compared to control (P[lt]0.05). Further, exposure to BPA at 25 [micro]g/kg and maintenance on HFD induced oxidative stress in Leydig cells as indicated by increased generation of reactive oxygen species (ROS) compared to the NFD control group (P[lt]0.05). Interestingly, antioxidant enzymes were differentially affected by the interaction of BPA and HFD. For example, superoxide dismutase activity was decreased in Leydig cells from all HFD animals (P[lt]0.05), whereas catalase activity was up-regulated by exposure to BPA regardless of diet (P[lt]0.05). The results were replicated in vitro because incubation of Leydig cells with BPA (10 nM, 18 h) increased ROS production compared to control (P[lt]0.05). In addition, culture of BPA-free Leydig cells with hydrogen peroxide (0, 25, 50 [micro]M; 18 h), a prototype ROS generator, decreased testosterone (T) production (P[lt]0.05). Collectively, results indicate that BPA exposure and/or maintenance on HFD induces oxidative stress. Therefore, BPA disruption of Leydig cell T secretion may be due, in part, to increased ROS levels.[br][br]Sources of Research Support: This study was supported in part by NIH ES-015886.[br][br]Nothing to Disclose: MKN, MA, MD, FFB, RLJ, BTA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1974 50 168 SAT-47 PO35-01 Saturday 107 2012


108 ENDO12L_SAT-48 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Activin Signaling Regulates Sertoli Cell Differentiation and Function Peter K Nicholls, Peter G Stanton, Justin Chen, Paul Gregorevic, Craig A Harrison Prince Henry[apos]s Institute of Medical Research, Clayton, Australia; Baker IDI Heart and Diabetes Institute, Melbourne, Australia Throughout development and in early postnatal life, proliferating Sertoli cells of the testis direct the formation of the seminiferous epithelium. At puberty, Sertoli cells enter a [apos]terminally[apos] differentiated state, characterised by the establishment of a functional blood testis barrier (BTB), the ability to sustain spermatogenesis, and withdrawal from mitosis. Activin A, a member of the transforming growth factor-[beta] (TGF-[beta]) superfamily, is a key regulator of Sertoli cell proliferation during embryogenesis, and a decrease in activin signalling corresponds with terminal differentiation. In this study, we sought to understand the function of activin upon differentiated Sertoli cells using an in vivo model of elevated systemic activin A. Chronic activin signalling reduces testis mass and induces a cumulative testicular dysgenesis phenotype in adult mice, consistent with a failure of Sertoli cells to support spermatogenesis. Activin was found to potently disrupt BTB function in adult mice, and tight junction formation in differentiated 20 dpp rat Sertoli cells. Furthermore, manipulation of activin signalling re-initiates a program of cellular proliferation in rat Sertoli cells. Proliferative cells were characterised by reduced expression of mature markers (including clusterin [0.32-fold, p[lt]0.01] and claudin-11 [0.23-fold, p[lt]0.01]), and re-expression of juvenile markers (including cytokeratin-18 [18-fold, p[lt]0.01] and ID3 [3.5-fold, p[lt]0.01]) following activin exposure. Our results confirm that Sertoli cells maintain the capacity to proliferate beyond puberty and show that activin promotes a functional de-differentiation of rodent Sertoli cells. This study indicates that activin signalling must be strictly controlled in the adult in order to maintain Sertoli cell function in spermatogenesis.[br][br]Sources of Research Support: National Health and Medical Research Council of Australia Project Grants (#1006488 and #1009144).[br][br]Nothing to Disclose: PKN, PGS, JC, PG, CAH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1454 50 169 SAT-48 PO35-01 Saturday 108 2012


109 ENDO12L_SAT-49 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Estrogen Receptors and Sertoli Cell Function Thais FG Lucas, Catarina S Porto, Maria de Fatima M Lazari Escola Paulista de Medicina - UNIFESP, S[atilde]o Paulo, Brazil Aim: Sertoli cells play a key role in the control of germ cell development, and are important targets for androgen and estrogen actions. In rat Sertoli cells, 17beta-estradiol (E2) activates translocation of estrogen receptors (ESRs) to the plasma membrane, and activates ERK1/2 phosphorylation (Biol Reprod 78:101, 2008). Expression of Cyclin D1 is increased by E2 and the ESR1-selective agonist PPT, but not by the ESR2-selective agonist DPN, indicating the role of ESR1 in Sertoli cell proliferation (Biol Reprod, 2012). In mammals, progression through the cell cycle is regulated by the association of Cyclins and Cyclin-dependent kinases (CDK), and the activity of these complexes is inhibited by p27Kip1 (reviewed in Genes Dev 18:2699, 2004). The role of E2 in the regulation of p27Kip1 has not been explored. Moreover, gene expression involved in differentiation, hormone synthesis and junction proteins in Sertoli cells may be regulated by GATA-1 transcription factor (reviewed in Mol Endocrinol 22:781, 2008). GATA-1 expression in Sertoli cells is not regulated by androgen (Endocrinology 146:2674, 2005), but the role of E2 has not been explored. We now report the regulatory role of E2, PPT and DPN on the expression of p27kip1 and GATA-1 in rat Sertoli cells. Methods and Results: Primary culture of Sertoli cells was obtained from 15-day old Wistar rats. Cells were incubated in the absence (control) and presence of E2 (0.1 nM), PPT (10 nM), or DPN (10 nM) for 24 hours at 35[deg]C. Cells were untreated or pretreated with the MEK inhibitor U0126 (20 [mu]M), the PI3K inhibitor wortmannin (1 [mu]M), the disruptor of the CREB:CBP complex KG-501 (25 [micro]M), or NFkB1 nucleus translocation inhibitor SN50 (1 [micro]M). Afterwards, cells were stimulated with E2, PPT or DPN, and the expression of p27kip1 and GATA-1 was determined by Western Blot. E2 or DPN increased the expression of p27kip1 and GATA-1 (2.3 and 1.5-fold, respectively), whereas PPT had no effect. Wortmannin or KG-501 blocked the effects of E2 or DPN on the expression of p27kip1 and GATA-1, while U0126 and SN50 had no effect. Conclusion: 17beta-estradiol modulates Sertoli cell proliferation through ESR1-mediated increase of cyclin D1 and ESR2-AKT-CREB-mediated increase of p27kip1. Possible effects of E2-ESR2 on gene transcription may be mediated by increased expression of GATA-1. The present study reinforces the important role of estrogen for normal testis development and homeostasis.[br][br]Sources of Research Support: FAPESP and CNPq.[br][br]Nothing to Disclose: TFGL, CSP, MdFML 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 927 50 170 SAT-49 PO35-01 Saturday 109 2012


110 ENDO12L_SAT-50 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Activation of PPAR[alpha] and PPAR[beta]/[delta] Regulates Sertoli Cell Metabolism Mariana Regueira, Maria Fernanda Riera, Silvana Lucia Artagaveytia, Maria Noel Galardo, Eliana Herminia Pellizzari, Selva Beatriz Cigorraga, Silvina Beatriz Meroni Hospital de Ni[ntilde]os R Guti[eacute]rrez, Buenos Aires, Argentina Sertoli cells (SC) provide the structural and nutritional support for germ cell development. SC actively metabolizes glucose and converts it to lactate, which is the major energy source for germ cells. On the other hand, it has been postulated that SC uses fatty acids (FA) as a source of energy. The transcription factors PPAR[alpha] and PPAR[beta]/[delta] [mdash]members of the PPARs nuclear receptor family[mdash] participate in the expression of genes involved in FA metabolism in many cells types. The aim of this work was to study the participation of PPAR[alpha] and PPAR[beta]/[delta] activation in the regulation of genes involved in FA metabolism and in lactate production in SC. Cultures of SC obtained from 20-day-old rats were incubated for different periods of time without (control) or with variable doses of WY14643 (WY) or GW0742 (GW) [mdash]pharmacological activators of PPAR[alpha] and PPAR[beta]/[delta] respectively. Carnitine-palmitoyltransferase 1 (CPT1) mRNA levels were detected by Northern Blot and long- and medium-chain 3-hydroxyacyl-CoA dehydrogenases (LCAD, MCAD) and the fatty acid transporter CD36 (FAT/CD36) mRNA levels were analyzed by RQPCR. Results expressed as fold-increase in mRNA levels combining three independent experiments (mean[plusmn]SD), obtained in 48h incubations with 10[micro]M WY and 5[micro]M GW are shown; [bold]CPT1[/bold] WY: 2.3[plusmn]0.6*; GW: 3.3[plusmn]0.2*; [bold]LCAD[/bold] WY: 1.9[plusmn]0.4*; GW: 2.8[plusmn]0.3*; [bold]MCAD[/bold] WY: 1.9[plusmn]0.3*; GW: 1.8[plusmn]0.4* and [bold]FAT/CD36[/bold] WY: 2.5[plusmn]0.6*, GW: 2.7[plusmn]0.5* (*p[lt]0.05 vs. control). While PPAR[alpha] activation had no effect on lactate production, PPAR[beta]/[delta] activation increased it (Control: 7.9[plusmn]0.5; GW: 11.4[plusmn]0.6[sup]# [/sup][mu]g/[mu]gDNA,[sup] #[/sup]p[lt]0.001 vs. control). Phosphorylation of the Pyruvate Dehydrogenase Complex (PDC) [mdash]enzymatic complex that regulates pyruvate to acetylCoA flow[mdash] was also evaluated by Western Blot. While an increase in phospho-PDC by PPAR[beta]/[delta] activation was observed, PPAR[alpha] activation had no effect. It is suggested that the increased phosphorylation of PDC by PPAR[beta]/[delta] activation, which results in inhibition of enzymatic activity, may be responsible for an increase in pyruvate availability that is converted to lactate. Altogether these results suggest that in SC, PPAR[alpha] and PPAR[beta]/[delta] participate in the regulation of FA metabolism. PPAR[beta]/[delta] also regulates lactate production and phosphorylation of PDC, which suggests a participation of this transcription factor in the provision of lactate for germ cell development.[br][br]Sources of Research Support: PIP2008N[ordm]806; PICT2007N[ordm]1004.[br][br]Nothing to Disclose: MR, MFR, SLA, MNG, EHP, SBC, SBM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 556 50 171 SAT-50 PO35-01 Saturday 110 2012


111 ENDO12L_SAT-51 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Evidence of Defective Release of Testosterone from XXY Mouse Testis Iqbal Munir, Jeffrey Kim, Lue YanHe, Leung Andrew, Christina Wang, Swerdloff Ronald Harbor-UCLA Medical Center, UCLA School of Medicine, Torrance, CA [bold]Objective: [/bold]In an experimental XXY mouse model of Klinefelter Syndrome, we have previously demonstrated that low peripheral serum testosterone (T) and elevated gonadotropins are associated with hyperplasia and hypertrophy of Leydig cells. Isolated Leydig cells from XXY testis produce significantly higher amounts of T [italic]in vitro[/italic] than XY controls. The aim of the study was to further characterize T production in XXY testis and to find direct evidence of a defect in the release of T to the circulation in XXY mice.[br][bold]Methods: [/bold]XXY mice were generated in our lab as published before. Testes of 7 adult XXY mice and their 7 XY littermates were used in the experiments. Testicular venous system was ligated and blood collected from the venous effluent. Testes were dissected out, decapsulated and incubated in the serum free medium in the presence or absence of 10 ng/ml of lutenizing hormone (LH) for 3 h. T was measured in the incubation medium, testicular homogenate and testicular venous effluent using LC-MS/MS. Expression of androgen binding protein (ABP) was evaluated in freshly isolated testicular homogenate by Western blot.[br][bold]Results: [/bold]Incubation of decapsulated whole testis in culture medium showed higher testicular content of T in XXY mice (25 [plusmn] 4.3 ng/testis, mean [plusmn]SEM,) in comparison to XY (17[plusmn] 2.6 ng/testis). Addition of LH to the culture medium further increased T content in XXY animals (40 [plusmn] 8.3 ng/testis) and XY testis (35 [plusmn] 7.3 ng/testis). There was no difference in the amount of T released in the medium from XY vs. XXY testis. T concentration in the testicular venous effluent, was 3-fold lower in the venous effluent from XXY testis (6.4 [plusmn] 0.85 ng/ml) compared to XY testis (18.3 [plusmn] 3.4 ng/ml, p=[lt]0.05). Western blot analysis demonstrated two subunits of ABP approximately 48 and 38 kDa. The 48 kDa subunit expression was higher in XXY testis whereas 38 kDa subunit had higher expression in XY testis.[br][bold]Conclusions: [/bold]Our data provide further evidence that synthesis of T is not impaired in the XXY testis. The amount of T is not lower in the testis of XXY mice but there is a defect in the release of T from testis to the venous circulation as shown by the lower concentration of T in testicular venous effluent. The similar concentrations of T in the culture media of decapsulated XY and XXY testes reflect leakage from the interstitial space. Differences in the ABP subunits in XY vs XXY testis may play a role in sequestration of T in mouse model XXY testes.[br][br]Nothing to Disclose: IM, JK, LY, LA, CW, SR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1543 50 172 SAT-51 PO35-01 Saturday 111 2012


112 ENDO12L_SAT-52 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Expression and Role of FoxO3a in the Testis Young-Suk Choi, Eun Jig Lee Yonsei University College of Medicine, Institute of Endocrine Research, Seoul, Republic of Korea; Yonsei University College of Medicine, Seoul, Republic of Korea; Northwestern University Feinberg School of Medicine, Chicago, IL Infertility was positively correlated with age in female FoxO3a KO mice. Degeneration in early proliferation of granulose cell was found. This study focuses mainly on the roles of FoxO3a in testis. The expression of FoxO3a starts from the embryonic day 13.5 and this lasts until the post pubertal week 3 (PPW3) in testicle. After PPW3, most of the expression of FoxO3a occurs in the nucleus of Leydig cell, but after PPW5, FoxO3a occurs both in the nucleus and cytoplasm. When GnRH signaling is defected in 5 weeks old male C57/BL6 mouse, FoxO3a was increased in the nucleus. When LH is treated on R2C cell, there was increase in phosphorylation of FoxO3a by AKT that is indicating LH regulates FoxO3a localization.[br]When active form of FoxO3a-TM adenovirus was introduced to R2C cell, a Leydig tumor cell, the concentration of testosterone hormone and StAR protein was reduced. Also when FoxO3a and StAR promoter-Luc vector were co-transfected into FT293 cell for reporter assay, FoxO3a inhibited StAR promoter activity. In conclusion, FoxO3a is a modulator of LH signal and plays an important role in testosterone production in the testis.[br][br](1)Murayama C., et al., Mol Cell Endocrinol 2012; 350(1): p. 1-9. (2)IHong Z.Y., et al,Cancer Lett 2012; 314(1): p. 34-40. (3)Fukuda S., et al.,J Ovarian Res, 2009. 2(1): p. 17.[br][br]Nothing to Disclose: Y-SC, EJL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 785 50 173 SAT-52 PO35-01 Saturday 112 2012


113 ENDO12L_SAT-53 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Gonadotroph Responsiveness to PACAP Signaling through the Short and HOP1 Isoforms of the PAC1 Receptor Rong Q Yang, Kimberly E Leake, Stephen J Winters, Joseph P Moore University of Louisville School of Medicine, Louisville, KY; University of Louisville School of Medicine, Louisville, KY There is increasing evidence that PACAP plays a role in the development and regulation of gonadotroph function. PACAP signals through VPAC1 and VPAC2, two G-protein-coupled receptors that are also activated by VIP, and through the PACAP specific receptor, PAC[sub]1[/sub]R. There are multiple splice variants of PAC[sub]1[/sub]-R (1). The Short and the Hop1 isoforms are expressed in gonadotrophs and act through the G[alpha]s pathway, while the Hop1 isoform also stimulates the G[alpha]q pathway to increase intracellular calcium (2). We sought to determine which PAC[sub]1[/sub]-R subtype is most influential in PACAP activation of the PACAP promoter and the follistatin promoter, a key PACAP target that mediates its effect to reduce FSH[beta] and GnRH-R expression. L[beta]T2 gonadotroph cells were transfected with 50 ng of either Short or HOP1 receptor expression vector and with a PACAP or follistatin promoter - luciferase construct. Transfected cells were treated for 6 h with increasing doses of PACAP, and luciferase activity was quantified. Over-expression of either Short or Hop1 increased follistatin promoter activation by PACAP equally. Mutation of the AP1 or CRE sites in the follistatin promoter reduced PACAP activation by the Short receptor only. Overexpression of either receptor increased PACAP promoter activation but the Hop1 receptor was more effective. Mutation of the proximal CRE of the PACAP promoter abolished stimulation. To identify genes activated by PACAP, L[beta]T2 cells were transfected with control or HOP1 vector and cultured with or without 10 nM PACAP for 6 h. In both groups, PACAP stimulated genes associated with PKA and p53 pathways and TRK receptor signaling equally and decreased DAR2 signaling molecules. Signaling pathway molecules stimulated more robustly in HOP1 transfected cells included PI3K/AKT, ERK/AKT, NGS and BMP pathways. Signaling pathway molecules stimulated only in HOP1 transfected cells included IP3 metabolism, calcium signaling, PKCb, JAK/STAT and NO pathways and receptor signaling molecules associated with GnRH, PPARa/RxRa, VEGF, TGFb, EGF, IGF-1, AR, ER, GR, and Wnt/b-catenin. Some of the genes significantly altered by PACAP exposure and relevant to gonadotroph function include increases in INHA, cMyc, junD, SMAD4, LIMHX1, LEP and ACVR2B and decreases in INHBB, CREBBP, ACVR1, and EGR1. Signaling through the HOP1 receptor likely involves cross-talk with multiple signaling pathways and may modulate gonadotroph responses to multiple endocrine and paracrine factors.[br][br](1) Vaudry D et al., Pharmacol. Rev. 2009; 61:283-357. (2) Mustafa T et al., J. Biol. Chem. 2007; 282:8079.[br][br]Sources of Research Support: NIH R01-HD050571 awarded to JPM. NIH Summer Endocrine Research Training Program Grant T35 DK072923.[br][br]Nothing to Disclose: RQY, KEL, SJW, JPM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2173 50 174 SAT-53 PO35-01 Saturday 113 2012


114 ENDO12L_SAT-54 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Comparative Expression of the Androgen Receptor in Immature and Mature Testes from Monkey and Human Espernaza Berensztein, Tony Plant, Roberto Ponzio, Diego Chirico, Marco A Rivarola, Alicia Belgorosky Hospital de Pediatria Garrahan, Buenos Aires, Argentina; Pittsburgh University, Pittsburgh, PA; Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina The androgen receptor (AR) is highly expressed in Sertoli cells (SC) of the adult human testis (HT) but not during infancy when expression of this receptor is found primarily in peritubular (PC) and interstitial cells (IC) (Berensztein et al. Pediatr Res 2006). In human infancy, serum gonadotropins and testosterone levels reach pubertal values but spermatogenesis is not initiated suggesting that FSH stimulation at this stage of development is not able to induce AR expression in SC. To analyze the role of gonadotropins in the regulation of the expression of testicular AR we have analyzed 1) the ontogenesis of AR expression by immunohistochemistry both in HT and in a higher primate model, the Macaca mulatta testis (MT),and 2) the effect of [italic]in vivo[/italic] LH and FSH stimulation on AR expression in SC in juvenile MT. RESULTS. 1) In both HT and MT, a similar cellular distribution of AR was found in all age groups: AR was positive in Leydig cells (LC), IC, PC and SC, and negative in germ cells (GC). In normal HT, the % of SC expressing AR (%AR) in Neonates ([lt] 1 month (m) old, n=6) was 2.25%, and in Infants (1-7 m, n=10) 2.96%, values significantly lower than in Prepubertal (5.9%,1-11 years, n=8) (p[lt]0.05) and in Pubertal (92.3.1%,14-15 y, n=4) subjects; in the adult subjects, it was 100%. Moreover, a significant positive linear regression of % AR as a function of age was found (r=0.951, p[lt]0.001). In MT, % AR in Neonates (9-18 d old, n=2) was 0.5; in Infants (4-5 m, n=2) 0, values significantly lower than in Juvenile (51%, 13-36 m, n=5) and Pubertal animals (86.1%, 42-48m, n=2); in adults (60 m, n=4) %AR was 96.7%. 2) Gonadotropin stimulation for 11 days of juvenile MT induced AR expression in SC. Specifically, %AR during treatment with FSH (79.3 [plusmn]13.6), LH (69.2 [plusmn]13) and FSH+LH (85.5[plusmn] 10.7) was significantly greater, p[lt]0.05 (Bonferroni test) with respect to that during vehicle (35.5[plusmn]17.6 %). Taken together, these results suggest 1) that upregulation of AR expression in SC during the transition from the neonatal to juvenile phase of development in primates is relatively gonadotropin independent, and 2) that LH and FSH appear to be equally important in driving the gonadotropin dependent upregulation of AR expression at puberty. Finally, validating the nonhuman primate model of Macaca mulatta for the human situation opens a range of possibilities to enrich our current knowledge of the postnatal development of HT.[br][br]Sources of Research Support: NIH grant,U54 HD 08160 (supported the generation of the monkey tissue).[br][br]Nothing to Disclose: EB, TP, RP, DC, MAR, AB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 185 50 175 SAT-54 PO35-01 Saturday 114 2012


115 ENDO12L_SAT-55 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Involvement of AMPK in Testicular Function Pauline Tartarin, Edith Guibert, Aminata Toure, Marc Foretz, Joelle Dupont, Benoit Viollet, Pascal Froment INRA, Nouzilly , France; Inserm, U1016, Institut Cochin; Cnrs UMR8104; Univ Paris Descartes, Sorbonne Paris Cit[eacute], Paris, France The 5 [apos]AMP-activated protein kinase or AMPK is a key protein kinase in the regulation of cell metabolism. It is also an important indicator of energy reserves. AMPK is the target of a drug, metformin, frequently used to treat diabetes and also certain female infertility associated with insulin resistance (syndrome polycystic ovaries). However few studies investigated the role of metformin and its target, AMPK, on male fertility. Thus, we analysed the involvement of this kinase by using 1/a male mouse line inactivated for the catalytic alpha 1 subunit of AMPK, 2/wild type males newborn mice from females treated with metformin during the first half of gestation.[br]Inactivation of AMPK led to a decrease in fertility associated with hyperandrogenism of testicular origin. Strangely, no modification of the testis structure was observed: neither in testis size or cell composition nor in apoptosis or in proliferation. However, sperm alpha1 AMPK-/- showed a lack of mobility in association with head structural anomalies, compared with male controls. Mitochondria in alpha1 AMPK-/- spermatozoa present also alterations that could be a cause of the lower mobility. In addition, hyperandrogenia was linked to hyperactivity in Leydig cells. Gonadotropins secretion were not modified in alpha1 AMPK-/- mice.[br]On the other hand, administration of metformin (an activator of AMPK) to pregnant mice showed a reduction in the testicular volume, diameter of the seminiferous tubules and the number of Sertoli cells. During administration of metformin to mothers, the intratesticular testosterone concentration in male fetuses decreased by about 30% compared to male fetuses from control mothers.[br]These two approaches suggest a role of AMPK in the testicular control (during testis development, sperm and testoterone production) similarly to its action in ovarian functions (oocyte quality and steroid production).[br][br]Sources of Research Support: This work was supported by the Agence National pour la Recherche (Fertinergy project).[br][br]Nothing to Disclose: PT, EG, AT, MF, JD, BV, PF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1909 50 176 SAT-55 PO35-01 Saturday 115 2012


116 ENDO12L_SAT-56 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Distinct Expression Patterns Predict Differential Roles of miRNA-Binding Proteins, Lin28 and Lin28b, in the Mouse Testis Francisco Gaytan, Susana Sangiao-Alvarellos, Maria Manfredi-Lozano, David Garcia-Galiano, Antonio Romero-Ruiz, Silvia Leon, Francisco Ruiz-Pino, Fernando Cordido, Leonor Pinilla, Manuel Tena-Sempere University of Cordoba, Cordoba, Spain; University of A Coru[ntilde]a, A Coru[ntilde]a, Spain Lin28 (also termed Lin28a) and Lin28b are related RNA-binding proteins, involved in the control of microRNA maturation, especially of the let-7 family, with putative roles in the regulation of stem cell differentiation and early (embryo) development. However, their function in adult tissues and during postnatal maturation remains ill defined. Despite the general assumption that Lin28 and Lin28b share similar targets and overlap in terms of function, conclusive demonstration of such a redundancy in different tissues and cell types is still missing. Recently, expression of Lin28 has been reported in the rodent testis, and human genomic studies have disclosed the association between variations in or around the LIN28B locus and the age of menarche; observations that suggest putative roles of Lin28 proteins in mammalian reproduction, whose mechanisms are yet to be defined. We document herein the pattern of RNA expression and protein distribution of Lin28 and Lin28b in the mouse testis during postnatal development and in adulthood. Expression of Lin28 and Lin28b mRNAs was detectable in mouse testis across postnatal maturation, from neonatal to adult periods. Of note, Lin28 peptides displayed totally different patterns of cellular distribution: Lin28 protein was prominently located in undifferentiated and type-A1 spermatogonia within the seminiferous tubules, whereas Lin28b was confined to interstitial Leydig cells and spermatids. These differential patterns of distribution were conserved at earlier stages of postnatal maturation. Moreover, relative mRNA levels of Lin28 and Lin28b disparately varied between neonatal and pubertal periods, with peak levels of Lin28 in the infantile testis and sustained elevation of Lin28b mRNA in young adult male gonads; an age when relative levels of let-7a and let-7b miRNAs were significantly suppressed. Altogether, our data are the first to document the divergent patterns of cellular distribution and mRNA expression of Lin28 and Lin28b in the mouse testis along postnatal maturation; a phenomenon which is suggestive of distinct functional roles of these two related, but apparently not overlapping, miRNA-binding proteins in the male gonad.[br][br]Sources of Research Support: BFU2011-25021 (Ministerio de Economia y Competitividad, Spain).[br][br]Nothing to Disclose: FG, SS-A, MM-L, DG-G, AR-R, SL, FR-P, FC, LP, MT-S 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1427 50 177 SAT-56 PO35-01 Saturday 116 2012


117 ENDO12L_SAT-57 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) A Specific Region in the 5[prime] Flanking Region of Gonadotropin-Regulated Testicular RNA Helicase (GRTH/DDX25) Gene Directs Its Cell-Specific Expression in Testicular Germ Cells Raghuveer Kavarthapu, Chon-Hwa Tsai-Morris, Masato Fukushima, Joaquin Villar, James Pickel, Maria L Dufau National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD GRTH is a multifunctional protein post-transcriptional regulator of genes that is essential for round spermatid elongation and completion of spermatogenesis. This helicase also prevents the Leydig cells (LC) from overstimulation of gonadotropin-induced androgen pathway by promoting degradation of StAR protein. In transgenic mice (Tg) carrying deletions of the GRTH 5[apos] flanking regions upstream of the promoter domain with GFP as the reporter gene, we previously demonstrated that the 205 bp promoter of the gene contains the necessary elements for LC specific expression. No expression in germ cells (GC) was found when Tg mice carrying the 5[apos] sequence extended to 3.6 kb, and 1.4 kb 5' to the ATG codon contains a functional half-site androgen response element. To further define regulatory regions of GRTH required for expression in GC during development, Tg mice were generated carrying 5[apos] flanking sequence 6.4 kb (6.4 kbTg) and/or various deletions. Within the 3.6-6.4 kb region, similar clusters of other known sequences that direct GC specific expression were identified. Immunohistochemistry showed that 6.4 kbTg directed GFP expression in both GC and LC. Deletion of the sequence between 1.4 kb to 3.6 kb (6.4-3.6 kb/1.4 kb Tg) direct GFP expression in GC only. In these lines, GFP expression was found to be stage specific in spermatocytes, round spermatids and elongating spermatids. Maximal expression of GFP in spermatocytes was observed in the pachtyene spermatocytes entering metaphase of meiosis. No expression was found in Sertoli cells. Western blots confirmed the GPF expression in all of these Tg lines. Our studies have generated and characterized transgenic lines that can be utilized to define basal and hormonally regulated transcription of the GRTH gene in a cell specific manner, and to further advance our knowledge on the regulation of GRTH action in male reproduction.[br][br]Sources of Research Support: The IRP, NICHD.[br][br]Nothing to Disclose: RK, C-HT-M, MF, JV, JP, MLD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1937 50 178 SAT-57 PO35-01 Saturday 117 2012


118 ENDO12L_SAT-58 POSTER SESSION: Regulation of Testicular Function (1:30 PM-3:30 PM) Expression and Gene Regulation of ARR19-[beta] in Mouse Testis Hana Kang, Keesook Lee Chonnam National University, Gwangju, Republic of Korea ARR19s, also known as Cmtm2 and cklfsf2, are MARVEL transmembrane domain containing proteins and characterized as two isoforms, ARR19-[alpha] and ARR19-[beta], with [sim]50% identity of amino acids in mice. ARR19-[alpha], which is highly expressed in the testis, was recently shown to be a factor regulating the function of testicular Leydig cells. In this study, we investigated the expression of ARR19-[beta] and its gene regulation in the testis. Besides highly expressed in the testis, ARR19-[beta] was also weakly expressed in other adult tissues such as epididymis, lung and kidney in mice. Further studies revealed that ARR19-[beta] expression was developmentally regulated in the testis. Immunohistochemical analysis of mouse testis showed that ARR19-[beta] protein was detected only in interstitial cells in young testis, whereas it was highly expressed in both haploid germ cells and interstitial Leydig cells in adult testis. ARR19-[beta] protein was also detected in epididymal sperms. In order to study the gene regulation of ARR19-[beta] in the testis, we analyzed 2 kb upstream sequence of the ARR19-[beta] promoter, which revealed the presence of three putative GATA-1 binding motifs. GATA-1 is the key regulator of ARR19-[alpha] expression in mouse Leydig cells. As well as GATA-1, CREB also markedly increased the activity of the ARR19-[beta] promoter. These findings demonstrate that ARR19-[beta] is the target gene of both GATA-1 and CREB. Interestingly, GATA-1 is repressed by cAMP, whereas CREB is activated by cAMP, in testicular Leydig cells. Therefore, ARR19-[beta] expression may be developmentally regulated by GATA-1 in young mice without LH/cAMP signaling and by CREB in adult mice with LH/cAMP signaling. Together, these findings suggest that ARR19-[beta] may be involved in the development and function of germ cells and Leydig cells in the testis.[br][br]Nothing to Disclose: HK, KL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 411 50 179 SAT-58 PO35-01 Saturday 118 2012


119 ENDO12L_SAT-59 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Estrogen Enhances Recovery of Fertility in Irradiated Rat Testis Wei Zhou, Connie C Weng, Marvin L Meistrich University of Texas MD Anderson Cancer Center, Houston, TX We previously showed in rats, in which irradiation had completely blocked spermatogonial differentiation, that testosterone (T) suppression with GnRH-antagonist and antiandrogen stimulated spermatogenic recovery and addition of estradiol (E2) accelerated this recovery. We now report for the first time, that addition of E2 enhances recovery of fertility in irradiated (6Gy) LBNF1 rats. 10 wk after irradiation, LBNF1 rats were treated with GnRH-antagonist and antiandrogen (Group XAF), or GnRH-antagonist and antiandrogen with addition of E2 (Group XAFE) for 6 wks. The rats were then allowed to recover for 6 wks without any hormonal suppression/treatment. For fertility recovery analysis, rats were mated with Sprague-Dawley female rats for 4 wks. Although condensed spermatids began to appear in testis by 22 wks after irradiation, in cauda epididymis the sperm were only observed at 26 wks after irradiation. At both time points, the sperm head counts were higher in XAFE group than in XAF group. By 26 wks after irradiation, 60% of the rats in XAFE group (n=5) recovered their fertility and for each fertile male, 1.7 litters of rats were born. The average litter size is 10.4, comparable to that of control normal rats (litter size=11.5). None of the rats in XAF group produced any pups. Testis histology analysis showed that by 26 wks after irradiation, although about 50% of the seminiferous tubules in rats of XAF group still exhibited spermatogenesis differentiation, around 1/3 of these differentiated tubules were abnormal, with missing layer(s) of differentiated germ cells or abnormal structure or apoptosis; while in XAFE groups, over 70% of the tubules exhibited spermatogenesis differentiation; in the 3 rats that had recovered fertility, over 90% of the tubules showed spermatogenesis recovery, and less than 10% of these tubules were abnormal. In XAF group, only 20% of the seminiferous tubules contained condensed spermatids by 26 wks after irradiation; while in XAFE group, about 60% of the tubules contained condensed spermatids. These data suggested that E2 not only enhances fertility recovery in irradiated rat, but also prolongs the normal spermatogenesis cycles after spermatogenesis induction by testosterone suppression. Further study and elucidation of the mechanism will help understand the role of estrogen in male spermatogenesis and may provide novel therapy methods for patients of azoospermia induced by cancer therapies.[br][br]Nothing to Disclose: WZ, CCW, MLM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 590 51 180 SAT-59 PO35-02 Saturday 119 2012


120 ENDO12L_SAT-60 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Contrasting FSH Signaling Actions during Sertoli Cell Proliferation and Differentiation Periods: Potential Regulation Via Differential Expression of G Protein Subunits Coupled to the FSH Receptor Ana P Jacobus, William H Walker Magee Womens Research Institute-School of Medicine-University of Pittsburgh, Pittsburgh, PA Sertoli cells receive, integrate, and transmit signals required for spermatogenesis. In the immature testes, Sertoli cells proliferate. During puberty, Sertoli cells stop proliferating and then differentiate so that they are able to support germ cell development. Sertoli cells proliferation and maturation are regulated by several hormones and growth factors; however, only follicle-stimulating hormone (FSH) stimulates both the proliferation of Sertoli cells immediately after birth and the differentiation of the cells during puberty. The extent of Sertoli cell proliferation and the timing of differentiation determines the final number of Sertoli cells and therefore, the number of germ cells that can be supported. To better understand the processes that control Sertoli cell development, we are investigating the mechanisms by which FSH signaling can be switched from promoting proliferation to differentiation. We hypothesize that the expression or activities of G protein subunits associated with the FSH receptor are altered during the proliferative and differentiation phases of the Sertoli cell development. Our preliminary data indicates that in proliferative Sertoli cells cultured from 5 day-old rats, FSH activates PI3K, AKT, c-Raf and Erk Kinase (MEK). These data suggest that FSH may act via G[beta][gamma] associated with G[alpha]i. In differentiated Sertoli cells isolated from 20 day-old rats, cAMP levels are elevated through FSH activation of G[alpha]s, leading to an increase of PKA activity and consequently c-Raf/MEK and AKT activities are decreased. These data suggest that FSH preferentially acts via G[alpha]s in differentiated Sertoli cells from 20 day-old rats. We will assay the levels and activities of the G protein subunits coupled to FSH receptor in 5 and 20-day-old Sertoli cells. We expect to find that the levels of G[alpha]i, G[alpha]s and their associated G[beta][gamma] subunits are altered during proliferation and differentiation such that FSH signaling results in opposing regulation of MAPK and AKT pathways during the two stages of development.[br][br]Sources of Research Support: CAPES post doctoral fellowship n0 0634/11-5.[br][br]Nothing to Disclose: APJ, WHW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1787 51 181 SAT-60 PO35-02 Saturday 120 2012


121 ENDO12L_SAT-61 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Simultaneous Insulin and Leptin Signaling in POMC Neurons Promotes Fertility and Metabolic Homeostasis in Male Rodents Latrice D Faulkner, Abigail R Dowling, Jennifer W Hill The University of Toledo, Toledo, OH; The University of Toledo, Toledo, OH Hypothalamic pro-opiomelanocortin (POMC) neurons in the arcuate nucleus are critical regulators of energy balance and glucose homeostasis. The POMC precursor is enzymatically processed to produce both the anorectic peptide [alpha]-melanocyte-stimulating hormone ([alpha]-MSH) and the endogenous opioid [beta]-endorphin. [beta]-endorphin exerts a continuous restrictive influence on GnRH secretion and the reproductive axis in both primates and rodents. Indeed, [beta]-endorphin immunoreactive terminals synapse on the GnRH neuron soma in these same species. Therefore, POMC neurons may function as critical nodes where metabolic and reproductive signals communicate.[br]Leptin and insulin convey information regarding energy stores to the CNS. POMC neurons contain both insulin and leptin receptors that regulate their activity. These regulatory signals also mediate the production of the pro-hormone convertases responsible for the post-translational processing of the POMC precursor. Here we demonstrate that male rodents lacking both insulin and leptin receptors in POMC neurons (LepR/IRPOMC mice) exhibit reduced fertility. Elevated testosterone levels, testis weights, and LH levels reflect this defect. We also examine whether the arcuate nucleus concentration of peptides produced from POMC processing and levels of pro-hormone convertases are affected. Our results suggest that the absence of leptin and insulin signaling in POMC neurons increases basal LH release and disrupts reproductive function via reduction in the inhibitory opioid tone on GnRH neurons. Thus, perception of leptin and insulin by POMC neurons is not only required for normal body weight and glucose homeostasis, but also to permit normal fertility. Additional studies are required to determine whether this action is mediated via direct or indirect neuronal connections.[br][br]Sources of Research Support: NIH Grant R00-HD-056491.[br][br]Nothing to Disclose: LDF, ARD, JWH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2007 51 182 SAT-61 PO35-02 Saturday 121 2012


122 ENDO12L_SAT-62 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Osteocalcin Mediates Insulin Regulation of Male Reproductive Functions Franck Oury, Mathieu Ferron, Huizhen Wang, Rajendra Kumar, Gerard Karsenty Columbia University, New York, NY; University of Kansas Medical Center, Kansas City, KS Genetics and biochemical evidence have established that bone, via the osteoblast-derived hormone osteocalcin, favors testosterone biosynthesis in the male mouse following its binding to GPRC6A on Leydig cells of the testis. The unraveling of this unexpected way of regulating male fertility raises the question as to know whether osteocalcin acts downstream of LH or instead defines a second pathway necessary for male fertility. Osteocalcin injection failed to correct any of the fertility defects in the pituitary Luteinizing hormone (LH)-deficient mice while in contrast Human chorionic gonadotropin (hCG) injection could correct the fertility defect in Osteocalcin-/- mice. Together these data indicate that osteocalcin does not act downstream of LH to regulate male fertility. Taking advantage of what is already known about osteocalcin regulation, we then showed through the analysis of cell-specific gene deletion and cell ablation mouse models that: (i) Insulin signaling in osteoblasts favors male fertility by modulating osteocalcin activity, (ii) bone resorption is the cellular mechanism used by insulin signaling in osteoblasts to enhance osteocalcin bioactivity. This study provides evidence that there is a direct link between energy metabolism and male fertility, and reveals that osteocalcin is necessary for this crosstalk to occur.[br][br]Nothing to Disclose: FO, MF, HW, RK, GK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2342 51 183 SAT-62 PO35-02 Saturday 122 2012


123 ENDO12L_SAT-63 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Cyclooxygenase 2 (COX2)/15d-PGJ2 System in Hamster Sertoli Cells: Its Modulation by FSH/Testosterone and Its Involvement on Glucose Uptake Maria Eugenia Matzkin, Eliana Herminia Pellizzari, Ricardo Saul Calandra, Selva Beatriz Cigorraga, Monica Beatriz Frungieri Institute of Biology and Experimental Medicine (IBYME-CONICET), Buenos Aires, Argentina; University of Buenos Aires, Buenos Aires, Argentina; Center for Endocrinological Investigations (CEDIE-CONICET), Buenos Aires, Argentina We have previously described a stimulatory effect of testosterone (T) on cyclooxygenase 2 (COX2) expression, prostaglandin (PG) synthesis as well as the involvement of PGs in the modulation of T production in hamster Leydig cells (1, 2). In the present study, we investigated the existence of a COX2/PGs system in hamster Sertoli cells (SC), its regulation by hormones (FSH and T) and the effect of one PG, 15d-PGJ2, on SC glucose uptake.[br]The Syrian hamster is a seasonal breeder. The exposure of adult hamsters to a short photoperiod (6h light/day) for 16 weeks triggers a drastic decrease in plasma levels of LH, FSH and T, as well as a severe testicular regression. SC were purified from testes of regressed adult hamsters and cultured under basal conditions for 48h. Subsequent incubation of SC in the presence of FSH (100 ng/ml), T (1 [mu]M) or the plasma membrane-impermeable T-BSA (1 [mu]M) for 1h significantly induced COX2 expression and MAPK1/2 phosphorylation (determined by Western blot) and 15d-PGJ2 production (quantified by immunoassay). The stimulatory effect of FSH and T-BSA on COX2 expression was abolished by the selective MEK1/2 inhibitor, U0126. In addition, T-BSA-induced COX2 expression and MAPK1/2 phosphorylation were reverted by the antiandrogen bicalutamide.[br]The presence of PPAR[gamma], the nuclear receptor for PGJ2 derivatives, was detected in hamster SC by RT-PCR and immunocytochemistry. 15d-PGJ2 (1 [mu]M) decreased the uptake of [sup]3[/sup]H-2-deoxyglucose (2DOG), a non-metabolizable glucose analogue, into SC. This effect was abolished by the PPAR[gamma] antagonist, BADGE. On the other hand, FSH, T and T-BSA increased 2DOG uptake, an effect further stimulated in the presence of meloxicam, a selective COX2 inhibitor. Thus, FSH and T regulate glucose uptake in hamster SC through, at least, two mechanisms, a direct positive modulation and an indirect negative effect exerted via 15d-PGJ2/PPAR[gamma].[br]In summary, these results demonstrate the presence of a COX2/PGs system in hamster SC and its up-regulation by FSH and T. Furthermore, results obtained in the presence of the plasma membrane-impermeable T-BSA suggest that T exerts a stimulatory effect on COX2/PGs through a non-classical mechanism that involves the presence of androgen receptors and MAPK1/2 activation. Overall, the COX2/15d-PGJ2 system might act as a local modulator of FSH and T actions on hamster SC function.[br][br](1) Matzkin ME et al., Reproduction 2009; 138:163. (2) Frungieri MB et al., Endocrinology 2006; 147:4476.[br][br]Sources of Research Support: Consejo Nacional de Investigaciones Cinetificas y Tecnicas (Conicet); Agencia Nacional de Promocion Cientifica y Tecnologica (ANPCyT); Facultad de Medicina, Universidad de Buenos Aires (UBACYT); Fundacion Roemmers.[br][br]Nothing to Disclose: MEM, EHP, RSC, SBC, MBF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 42 51 184 SAT-63 PO35-02 Saturday 123 2012


124 ENDO12L_SAT-64 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Impact of Aging and Obesity on Hepatic Sex Hormone-Binding Globulin Gene Expression in Women and Men Jyothi Gogineni, Marjan Karegar, Dushan T Ghooray, Charles R Scoggins, Stephen J Winters University of Louisville, Louisville, KY; University of Louisville, Louisville, KY Sex hormone binding globulin (SHBG) is a homodimeric glycoprotein produced by hepatocytes that transports testosterone and other steroids in human plasma, reduces their clearance, and regulates their entry into target cells. Multiple factors influence the plasma level of SHBG including genotype, sex, age, weight, diet, exercise, and a variety of hormones including thyroxine, growth hormone, cortisol, testosterone and estradiol. SHBG levels are low in obesity, and low levels beginning in childhood predict the development of the Metabolic Syndrome (MetS) and Type 2 diabetes (T2DM). However, the mechanisms that regulate SHBG are only partly understood. Experiments in HepG2 hepatocarcinoma cells and mice expressing an SHBG transgene reveal that the transcription factor HNF4[alpha] activates the SHBG promoter, COUP-TF antagonizes this effect, the peroxisome proliferator-activated receptor PPAR[gamma] represses transcription, and TNF[alpha] reduces SHBG expression partly through HNF4[alpha]. So far, however, no studies have examined SHBG gene expression in human liver. In this study, we obtained normal liver tissue from 25 patients (11F, 14M) age 46-82 yrs with colorectal cancer and ECOG performance status 0 who were undergoing partial hepatic resection as treatment for their disease. SHBG and HNF4[alpha] mRNA levels were measured by PCR, serum SHBG levels by ELISA, and clinical data were obtained. The BMI ranged from 16-34 in women and 16-37 kg/m[sup]2[/sup] in men. In agreement with previous results, serum SHBG levels were higher in women than men and diverged with increasing age, were markedly increased in women treated with estradiol or tamoxifen, and decreased with increasing BMI. SHBG mRNA levels were positively correlated to the plasma level of SHBG (r=0.37), were higher in subjects with lower BMI (r=-0.3), but were similar in men and women, and were unaffected by age. The expression level of HNF4[alpha] was highly positively correlated (r=0.81; p[lt]0.01) with the level of SHBG mRNA, and was also higher in thin patients than in those who were obese, but neither sex nor age predicted HNF4[alpha]. Moreover, while plasma SHBG was lower in those patients with MetS (n=9) or T2DM (n=8), SHBG mRNA and HNF4[alpha] levels were similar. Our results imply that the plasma level of SHBG is regulated by transcriptional and post-translational mechanisms, HNF4[alpha] is an important determinant of SHBG expression in human liver, and multiple factors contribute to the change in SHBG that occurs with aging or obesity.[br][br]Nothing to Disclose: JG, MK, DTG, CRS, SJW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 843 51 185 SAT-64 PO35-02 Saturday 124 2012


125 ENDO12L_SAT-65 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) GLUT3 and CASPR5 [mdash] Novel Genetic Factors in Male Infertility Alexander W Pastuszak, Carolina J Jorgez, Larry I Lipshultz, Dolores J Lamb Baylor College of Medicine, Houston, TX [bold]Introduction and Objective:[/bold] While several known genetic alterations cause male infertility, it is likely that many other currently unrecognized genetic etiologies exist. Here, we present genomic data implicating the glucose transporter GLUT3, as well as contactin-associated protein CASPR5, in male fertility.[br][bold]Methods:[/bold] Blood-derived genomic DNA from 22 men with non-obstructive azoospermia (NOA) and normal Y-chromosome microdeletion and karyotype analyses, as well as 4 fertile male controls, was used for array comparative genomic hybridization (aCGH) to assess for copy number variations (CNVs). Copy number gains and losses were identified and candidate fertility genes selected based on 1) the frequency of CNVs in a gene, 2) the magnitude of the gain/loss, and 3) available data on gene expression. CNVs were validated using Taqman qPCR, and DNA sequencing performed for genes of interest. Immunohistochemical staining of testis sections utilized commercially available antibodies and standard protocols.[br][bold]Results:[/bold] Copy number gains were identified using aCGH in GLUT3 in 2/22 NOA patients, and in CASPR5 in 1/22 patients and in no controls. These CNVs were confirmed in all patients using Taqman qPCR copy number assays (CNAs). Subsequent CNAs of DNA from 43 infertile men yielded 5 more men with GLUT3 gains, and one additional man with a loss in CASPR5. The frequency of GLUT3 CNVs in the general population (including fertile and infertile individuals) is approximately 5%, whereas our observed gain frequency is 16%. CNVs in CASPR5 are found in 0.002% of the general population and in 4% of our infertile male population. Sequencing of GLUT3 yielded SNPs in exons 2 (2465A[gt]C, 36K[gt]T), 6 (6505A[gt]G, 258L[gt]L), and 10 (14680C[gt]T, 436T[gt]T), none of which were predicted to be damaging. Staining for GLUT3 in testis of a male with 3 copies of GLUT3 and hypospermatogenesis demonstrated cytoplasmic Leydig cell and spermatocyte staining.[br][bold]Conclusions:[/bold] Copy number gains and losses in GLUT3 and CASPR5, respectively, were identified in several infertile males. Future work in the form of CASPR5 sequencing and subcellular localization and functional assays analyzing the impact of expression of both genes on fertility-related cell signaling pathways, as well as the study of animal models, will further elucidate the roles of these genes in male fertility.[br][br]Sources of Research Support: AUA Foundation Russell Scott, Jr., MD, Resident Research Award (AWP); NIH grants K12 DK0083014 (CJJ, DJL), the Multidisciplinary K12 Urologic Research (KURe) Career Development Program (CJJ, DJL), and by P01HD36289 from the Eunice Kennedy Shriver National Institute for Child Health and Human Development, NIH (DJL).[br][br]Nothing to Disclose: AWP, CJJ, LIL, DJL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1411 51 186 SAT-65 PO35-02 Saturday 125 2012


126 ENDO12L_SAT-66 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Circulating Endothelial Cells as Marker of Vascular Damage in Male Hypogonadism Domenico Milardi, Giuseppe Grande, Antonella Giampietro, Linda Tartaglione, Sara Palumbo, Francesca Vendittelli, Riccardo Marana, Alfredo Pontecorvi, Cecilia Zuppi, Ettore Capoluongo, Laura De Marinis Universit[agrave] Cattolica del S Cuore, Rome, Italy; Universit[agrave] Cattolica del S Cuore, Rome, Italy; Universit[agrave] Cattolica del S Cuore, Rome, Italy Testosterone (T) deficiency has become a frequently diagnosed condition in our current society with epidemic obesity and is associated with cardiovascular risk (1). Recent studies has established the importance of altered vascular endothelium function to cardivascular disease. The damage to the endothelium might also cause endothelial cell detachment, resulting in increased numbers of circultating endothelial cells (CECs) within the bloodstream (2).[br]To investigate if hypogonadism could modify CEC count in pherypheral bloodstream, we investigated CEC count by the CellSearch System in peripheral blood of twenty hypogonadic patients (25 to 55 years), studied after neurosurgical operation to remove nonsecreting pituitary adenoma or craniopharyngioma. The control group comprised ten age- and gender-matched healthy subjects. Antropomethric parameters, body mass index and waist circunference (WC) were evalutated. Serum total cholesterol, HDL cholesterol, triglycerides, glycosylated haemoglobin levels, T, estradiol, dihydrotestosterone, SHBG, LH and FSH were determined. CEC count was investigated by the CellSearch System, a semiauthomatic method to accurately and reliably enumerate CECs, sorted based on a CD146+, CD105+, DAPI+, CD45- phenotype (3).[br]CEC count/ml was increased in hypogonadic patients vs control group (mean[plusmn] SE 16.53[plusmn]7.55 vs 1.03[plusmn]0.14 cells/ml; p[lt]0.05). Correlation analysis in hypogonadic patients showed inverse correlation between T levels and CEC count/ml, positive correlation between waist circumference and CEC count/ml and inverse correlation between T levels and waist circumference. Regression analysis yielded a significant result for the model, with the highest adjusted R[sup]2[/sup] for T and WC, the most consistently significant variables.[br]Hypogonadism has been shown to be an indipendent determinant of endothelial disfunction, thus contributing to vascular pathology (4). The increase in CEC count in the group of hypogonadic patients represent a direct index of endothelial damage and could be a possible link between T deficiency and cardiovascular disease. WC in hypogonadal man may represent an adjuntive risk factor for endothelial damage and cardiovascular disease. Male hypogonadism induces endothelial disfunction. Visceral obesity may be implicated in the endothelial-disfunction induced by T deficiency.[br][br](1) Ullah MI et al., Horm Metab Res 2011; 43(3):153. (2) Blann AD et al., Thromb Haemost 2005; 93(2):228. (3) Strijbos MH et al., Thromb Haemost 2009; 102(2):347. (4) Foresta C et al., Clin Endocrinol 2008; 68(2):284.[br][br]Nothing to Disclose: DM, GG, AG, LT, SP, FV, RM, AP, CZ, EC, LDM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1924 51 187 SAT-66 PO35-02 Saturday 126 2012


127 ENDO12L_SAT-67 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Regenerated Luminal Epithelial Cells Are Derived from Pre-Existing Luminal Epithelial Cells in Adult Mouse Ventral Prostate Laura E Pascal, June Liu, Sudhir Isharwal, Daniel Metzger, Raquel Ramos Garcia, Jan Pilch, Susan Kasper, Karin Williams, Per Basse, Joel Nelson, Pierre Chambon, Zhou Wang University of Pittsburgh School of Medicine, Pittsburgh, PA; Universit[eacute] de Strasbourg, Coll[egrave]ge de France, Strasbourg, France; University of Cincinnati, Cincinnati, OH; University of Pittsburgh School of Medicine, Pittsburgh, PA Introduction: Determining the source of regenerated luminal epithelial cells in the adult prostate during androgen deprivation and replacement will provide insights into the origin of prostate cancer cells and their fate during androgen deprivation therapy. Prostate stem cells in the epithelial layer have been suggested to give rise to luminal epithelium. However, the extent of stem cell participation to prostate regrowth is not clear.[br]Methods: A recently developed genetic lineage tracing strategy was used to follow the fate of mouse prostatic luminal epithelial cells during androgen withdrawal and androgen replacement. To control for the specificity of prostatic luminal epithelial cell labeling and the time of labeling, a previously reported PSA[minus]CreERT2 transgenic mouse strain expressing tamoxifen (TAM)[minus]inducible CreERT2 recombinase driven by a 6[minus]kb human PSA promoter/enhancer was used. PSA[minus]CreERT2 mice were crossed with mT/mG mice to generate bigenic PSA[minus]CreERT2/R26[minus]EGFP mice. The luminal epithelial cells in intact prostate were genetically marked by transient administration of TAM, and the fate of marked luminal cells was determined during prostate regression and regrowth in the ventral prostate and compared to the fate of luminal epithelial cells previously determined in the lateral prostate.[br]Results: The PSA[minus]CreERT2[minus]based genetic lineage marking approach labeled fully[minus]differentiated luminal cells in the intact prostate, but not in the castrated prostate. In intact, non[minus]castrated PSA[minus]CreERT2/R26R[minus]GFP mice, [sim]70% of the luminal epithelial cells in the lateral prostate were labeled by GFP in the lateral prostate; and the percent labeling remained constant between intact and androgen manipulated mice. The efficiency of GFP genetic labeling by TAM was less in the ventral prostate than in the lateral prostate. Approximately 50% of the luminal epithelial cells were labeled by GFP in the ventral prostate, and the percent GFP labeling remained constant between intact and androgen manipulated mice. [br]Conclusions: Pre[minus]existing luminal epithelial cells were shown previously to be a source of regenerated luminal epithelial cells in the adult murine lateral prostate. This self-duplication of luminal epithelial cells was confirmed in the ventral prostate. Prostatic luminal epithelial cells could survive androgen deprivation and were capable of proliferating upon androgen replacement. This mechanism does not appear to be lobe specific.[br][br]Sources of Research Support: National Institutes of Health Grants 5 R37 DK51193, R01 CA 108675, 1 P50 CA90386, and P30 CA047904. LEP was a trainee of NIH T32 DK007774 training program and JL was a recipient of the Mellam Family Foundation Fellowship.[br][br]Nothing to Disclose: LEP, JL, SI, DM, RRG, JP, SK, KW, PB, JN, PC, ZW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 468 51 188 SAT-67 PO35-02 Saturday 127 2012


128 ENDO12L_SAT-68 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Steroidogenic Enzyme Expression in the Human Fetal Prostate Keely McNamara, Elana Rosman, Hiro Sasano, Yasuhiro Nakamura, Shen Yao, Ellen Shapiro, Hongying Huang, Alexander Kirschenbaum, Alice C Levine Tohoku University School of Medicine, Sendai, Japan; Mount Sinai School of Medicine, New York, NY; New York University School of Medicine, New York, NY; Memorial Sloan Kettering Cancer Center, New York, NY [bold]Background: [/bold]Androgens are necessary for normal prostate development. Recent reports demonstrate expression of steroidogenic enzymes in a subset of castration-resistant prostate cancer (PCa) cells and these findings have led to the development of new treatments for advanced disease. We previously demonstrated cell-type specific expression of type II 5-alpha-reductase (SRD5A2) in epithelial and mesenchymal cells of the human fetal prostate. We hypothesize that a subset of epithelial cells in the developing human prostate express steroidogenic enzymes involved in de novo androgen synthesis and that this expression in primary PCa may be a marker for more aggressive disease. [bold]Methods[/bold]: Tissues were obtained from 12 male fetuses (8-22 weeks gestation) at the time of pregnancy interruption and with the approval of the institutional review boards (Mount Sinai and New York University). All samples had been fixed in 10% formalin, embedded in paraffin and later deparaffinized and immuhistochemically stained for c17 and 17[beta]HSD5. Briefly, a citrate based (ph6) antigen retrieval step was used to retrieve the antigens. A methanol/H2O2 block was employed to quench endogenous peroxidase expression. Visualization of the antibody was carried out using the Histofine Kit (Nichirei, Japan) which is based on the principle of streptavidin-biotin amplification. Specificity of C17 staining was verified by antibody preabsorption. [bold]Results: [/bold]There was marked immunoreactivity of c17 in both ductal and acinar epithelial cells as in some stromal cells of 8, 9, 11, 12, 13, 16, 19, 21 and 22 weeks gestation human fetal prostates. There was also some weaker expression of 17BHSD5 in both stromal and epithelial cells of the same specimens. [bold]Conclusion:[/bold] Human fetal prostate epithelial cells express c17 as early as 8 weeks gestation in a subset of cells in the proximal ducts. These c17 positive fetal prostate epithelial cells may represent a progenitor population. PCa cells that similarly express steroidogenic enzymes may, therefore, represent a more primitive, potentially more aggressive subpopulation.[br][br]Nothing to Disclose: KM, ER, HS, YN, SY, ES, HH, AK, ACL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1598 51 189 SAT-68 PO35-02 Saturday 128 2012


129 ENDO12L_SAT-69 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Enhancement of the Antitumor Action of Calcitriol in Ectopic and Orthotopic Xenografts of Human Prostate Cancer Cells Harboring CYP24A1 shRNA in Mice Shaye Kamal Lewis, Julie N Stewart, Josephine B Addai, Hima V Vangapandu, Annisa L Lewis, Kristin Dittmar, Robia G Pautler, Dolores J Lamb Baylor College of Medicine, Houston, TX; Weil Cornell Medical College and The Methodist Hospital, Houston, TX; Weil Cornell Medical College and The Methodist Hospital, Houston, TX; Baylor College of Medicine, Houston, TX In many prostate cancer patients, androgen deprivation therapy results in a temporary reduction of tumor growth followed by androgen-independent tumor growth. Currently, there are no effective treatments for androgen-independent prostate cancer. Calcitriol inhibits the growth of cancer cells from multiple tissues including the prostate. An CYP24A1 is an important enzyme involved in the metabolism of calcitriol into a biologically inactive form. CYP24A1 is over-expressed during advanced stages of prostate cancer leading to calcitriol resistance. We tested the hypothesis that administration of low doses of calcitriol to DU145 prostate cancer cells stably expressing shRNA CYP24A1 will enhance inhibition of prostate cancer growth in xenograft mouse models. Tumors were produced by either intra-prostatic or subcutaneous injection of SCID or nude mice. Calcitriol or sesame oil (vehicle) was administered by oral gavage. MR imaging (Bruker Biospin Pharmascan 7.0T MRI) was done to assess intra-prostatic tumor growth. After 7 weeks of treatment, s.c. DU145 tumors stably expressing shRNA CYP24A1 receiving calcitriol showed a marked reduction in tumor volume compared to all experimental groups (150% difference in tumor volume compared with shRNA controls treated with calcitriol, p[lt]0.0001). Similarly, intra-prostatic tumor volume and proliferation decreased between 60 and 90 days of treatment in shRNA CYP24A1 xenograft-bearing mice treated with calcitriol. Metastasis occurred after orthotopic (38% of orthotopic engraftments, n=34; p[lt]0.0001, Fisher[apos]s exact test) but not s.c. injections. Local tumor invasion was predominantly observed beneath the urothelium of the bladder. Distant tumor dissemination was mostly located in the lymph nodes and diaphragm. Of importance, significantly fewer CYP24A1 shRNA xenograft-bearing animals treated with calcitriol (22%, n= 9; p= 0.0456) displayed metastatic tumors than did the other experimental groups. In both ectopic and orthotopic xenografts experiments, calcitriol treatments were not associated with any significant toxicity as evidenced by the absence of calcium phosphate deposits in the kidneys of all experimental groups. Thus, selective targeting CYP24A1 may facilitate the use of low doses of calcitriol to inhibit growth of prostate cancer cells while avoiding toxicity.[br][br]Sources of Research Support: This work was supported by W81XWH-07-1-0022 PC061154 Idea Development Award from the Department of Defense, Prostate Cancer Research Program Initiative.[br][br]Nothing to Disclose: SKL, JNS, JBA, HVV, ALL, KD, RGP, DJL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2235 51 190 SAT-69 PO35-02 Saturday 129 2012


130 ENDO12L_SAT-70 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Dicer1 Ablation in the Mouse Epididymis Causes Dedifferentiation of the Epithelium and Imbalance in Sex Steroid Signaling Ida Bjorkgren, Anton Krutskikh, Ilpo Huhtaniemi, Matti Poutanen, Petra Sipila University of Turku, Turku, Finland; University of Turku, Turku, Finland; Imperial College London, London, UK; University of Turku, Turku, Finland The epididymis, required for sperm maturation and storage, is a complex organ divided into several segments. Final differentiation of the epididymal epithelium takes place after birth and gives rise to a segment specific gene expression pattern. Recent studies on human and rat epididymides have shown that microRNAs (miRNAs) are differentially expressed at juvenile and adult stages, indicating a role for RNA interference (RNAi) in the postnatal development of the epididymis. To study the role of RNAi in epididymal development and function, we have generated a conditional knock-out (cKO) mouse line of Dicer1, the RNase III enzyme needed for processing pre-miRNAs to functional miRNAs. The lack of Dicer1 in the most proximal segments of the epididymis caused dedifferentiation of the epithelium with increased cell proliferation and a reduction in segment specific gene expression. The [italic] Dicer1 [/italic] cKO epididymis also presented with an imbalance in the expression of sex steroid receptors. Similar to androgen receptor [italic] (Ar) [/italic] cKO mice or mice with excess estrogen signaling, the expression of [italic] Ar [/italic] was downregulated while the expression of Estrogen receptor 1 increased in the most proximal segments of the [italic] Dicer1 [/italic] cKO epididymis. This suggests a role for the RNAi pathway as an important regulator of balanced estrogen-androgen action in the mouse epididymis.[br][br]Nothing to Disclose: IB, AK, IH, MP, PS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1192 51 191 SAT-70 PO35-02 Saturday 130 2012


131 ENDO12L_SAT-71 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Variants in Follicle-Stimulating Hormone Receptor Gene in Infertile Brazilian Men and the Correlation to FSH Serum Levels and Sperm Count Carla Peluso, Milton Ghirelli-Filho, Marcello Machado Gava, Denise Maria Christofolini, Caio Parente Barbosa, Bianca Bianco Faculdade de Medicina do ABC, Santo Andre, Brazil; Faculdade de Medicina do ABC, Santo Andre, Brazil In male, follicle-stimulating hormone (FSH) is essential for normal spermatogenesis and fundamental for Sertoli cell function and the induction and maintenance of qualitatively and quantitatively normal spermatogenesis by a specific receptor (FSHR). The aim of the study was to analyze the distribution of the follicle-stimulating hormone (FSH) receptor ([italic]FSHR[/italic]) Ala307Thr and Asn680Ser polymorphisms in infertile Brazilian men and evaluate the possible role of these polymorphisms on serum levels of FSH and in sperm count. A case-control study was performed comprising 138 infertile men with non-obstructive azoospermia (n=53) or severe oligozoospermia (n=85), and 217 fertile men as controls. Genotyping of [italic]FSHR[/italic] polymorphisms was performed by real time PCR. The results were analyzed statistically and a [italic]p[/italic]-value [lt]0.05 was considered significant. According to the sperm count, relatively similar [italic]FSHR[/italic] polymorphisms genotype and allele frequencies were found among the groups, and combined genotypes of two polymorphisms did not identify a haplotype associated with sperm count. Considering FSH serum level according to genotypes of the Ala307Thr and Asn680Ser polymorphisms individually, statistical analysis showed no difference among the groups. When the combined genotypes of the [italic]FSHR[/italic] polymorphisms were compared to FSH serum levels, no difference was also found among the groups. In conclusion, the findings demonstrate that, in Brazilian population studied, genetic variations, Asn680Ser and Thr307Ala, of the [italic]FSHR[/italic] gene are not correlated with serum FSH levels or sperm count in male infertility.[br][br]Sources of Research Support: This work was supported by grants from FAPESP (Funda[ccedil][atilde]o de Amparo a Pesquisa do Estado de S[atilde]o Paulo) #2011/08681-1.[br][br]Nothing to Disclose: CP, MG-F, MMG, DMC, CPB, BB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 8 51 192 SAT-71 PO35-02 Saturday 131 2012


132 ENDO12L_SAT-72 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Can Intratesticular Injection of Hypertonic Saline Replace Surgical Castration? Eun Young Jeon, Byung Kuk Kwak, Yong-Pil Cheon, Sung-Ho Lee Sangmyung University, Seoul, Republic of Korea; Sungshin Women[apos]s University, Seoul, Republic of Korea Recent evidence revealed that the intratesticular injection of hypertonic saline(20%) resulted in nadir testosterone level and sterile state in rats. To confirm the efficacy of this method in reproductive endocrine studies further, we investigated the anatomical changes and several target gene expression levels in the reproductive tissues. Single dose of hypertonic saline(750 [micro]l/testis) were administered into the testes of adult rats(5 months-old). Bilateral orchiectomy was performed at the same day of injection. Following 1, 2, 4 weeks and 3 months post-injection, the tissues were collected. The body weights were not changed significantly in both orchiectomy group and saline-injection group compared to those in intact group. The wet weights of reproductive tissues(testis, epididymis, seminal vesicle and prostate) and adrenal were significantly decreased in orchiectomy group and saline injected group from 4 weeks post-treatment when compared to those in intact group. The testes exerted slight atrophy and the tunica albuginea seemed to be intact in saline injected group. Histologically, however, larger parts of testicular tissue underwent necrosis and were barely recognizable after hematoxylin-eosin staining. Sloughing of immature germ cells from the basement membrane along with shedding cells in the intraluminal space was notable in most seminiferous tubules from the saline injected testis. In RT-PCR studies, the mRNA levels of hypothalamic GnRH and kisspeptin were significantly elevated in both orchiectomy group and saline injected group. Likewise, the expression levels of the subunits for pituitary gonadotropin(i.e. common alpha subunit, LH-beta and FSH-beta subunit) also were significantly increased. The present study confirmed that the direct injection of hypertonic saline into testis can induce a castration-like, testosterone-depriving effects on the accessory sex organs. Both chemically castrated and surgically castrated animals were almost identical not only in their histological features and but also in their expression patterns of hypothalamus-pituitary reproductive endocrine factors. Our findings suggest that this simple and inexpensive method would be a good replacement for the conventional orchiectomy and the hormone-based chemical castration(e.g. GnRH antagonists treatment).[br][br]Nothing to Disclose: EYJ, BKK, Y-PC, S-HL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1502 51 193 SAT-72 PO35-02 Saturday 132 2012


133 ENDO12L_SAT-73 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Determination of Peripheral Gonadal Hormones after the Treatment of Atypical Antipsychotics, Olanzapine and Risperidone in Wistar Male Rats Tulin Yanik, AK Mehmet, Denis Sezlev, Canan Kursungoz, Bulent Kurt, Ozan Akay, Suleyman Akarsu METU, Ankara, Turkey; GATA, Ankara, Turkey; GATA, Ankara, Turkey Sexual dysfunction is seen in approximately 30-80% of schizophrenic patients, which significantly reduces the quality of life. The antipsychotic treatment is one of the important causes of the sexual dysfunction in these patients. Two of the generally prescribed atypical antipsychotics, olanzapine and risperidone have less extrapyramidal side effects; although, they cause sexual and reproductive dysfunctions in males. The antipsychotics affect the levels of hypothalamus-pituitary-gonad (HPG) hormones that are mainly gonadotropin releasing hormone (GnRH), prolactin, follicle stimulating hormone (FSH), leutinizing hormone (LH) and also total testosterone which may lead to delay in spermatogenesis, decrease in sperm motility and quality, and cause morphological changes in testes. Atypical antipsychotics stimulate both dopaminergic and serotonergic systems and trigger release of LH, FSH, and testosterone. Therefore, we have investigated the peripheral levels of testosterone, LH, FSH and histological properties of the testes in the wistar-male rats (n=15) that were treated with olanzapine (4mg/kg/day) and risperidone (2mg/kg/day) orally for 4-wks. Plasma FSH, LH and testosterone levels were analyzed by ELISA. The plasma FSH levels were decreased by [sim]30% in the olanzapine treated group (1.47 mlU/ml) vs control (2.09 mlU/ml) and increased by [sim]35% in the risperidone group (3.20 mlU/ml [plusmn] 1.35 SEM) vs control. The plasma LH levels did not significantly change in both olanzapine and risperidone groups (0.55mlU/ml) vs control (0.53 mlU/ml). The plasma testosterone levels were also similar in all groups that were 0.43 mlU/ml in olanzapine and 0.69 mlU/ml risperidone groups vs control (0.61 mlU/ml). Although the animals[apos] testes were enlarged during the drug treatment, we did not detect any abnormalities in the diameter of seminifer tubules and their inflammation at the intertubular distance, Leydig cell morphology, as well. Finally, this slight change of FSH, a peptide evidenced to stimulate spermatogenesis, may render a beginning of change in the HPG axis during the atypical antipsychotic treatment.[br][br]Nothing to Disclose: TY, AKM, DS, CK, BK, OA, SA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2337 51 194 SAT-73 PO35-02 Saturday 133 2012


134 ENDO12L_SAT-74 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Analysis of Herpesvirus Infection in Human Infertility Mo Chen, Masashi Higuchi, Li-Yi Cai, Shun-ichiro Izumi, Takako Kato, Yukio Kato Meiji University, Kawasaki, Japan; Meiji University, Kawasaki, Japan; Meiji University, Kawasaki, Japan; Maternal and Child Health Hospital, Wuxi, China; Tokai University, Isehara, Japan We developed a transgenic (TG) rat using chimera gene composed of porcine FSHb promoter fused to the reporter gene, Human Herpesvirus1 Thymidine Kinase (HHV1-TK). TG rats show ectopic expression of HHV1-TK in the testes and exhibit male infertility. To clarify mechanism underlying the disruption of spermatogenesis in those TG rats is valuable to understand normal spermatogenesis. Recently we demonstrated an accumulation of HHV1-TK in the round spermatid of TG rat testis. In addition, we identified the ectopic transcription start site in the downstream of the translation start site of the HHV1-TK gene, resulting the production of truncated HHV1-TK. Observation that ectopic expression of HHV1-TK is due to an endogenous promoter/transcription site suggests us that HHV1 infected in human expresses its TK gene by ectopic promoter in testis resulting in accumulation of truncated HHV1-TK and this triggers male infertility as shown in HHV-TK TG rat. Hence, in this study, we have investigated a relationship between infection of herpes virus and human male infertility.[br]Semina were donated by Chinese male infertile patients (153 men, aged 21-49 years) with informed consent, followed by DNA preparation for PCR. Semen volume, sperm number and sperm mortality were examined. Nested-PCR was performed for four types of helpes virus. Amplified DNA was extracted after agarose gel electrophoresis and analyzed the nucleotide sequence[br]The virus DNAs of HHV1, HHV3, HHV5 and HHV6 were confirmed by PCR, electrophoresis and DNA sequencing and the virus infection was identified in 59 patients (39%). Number of patients infected were 39 (26%) for HHV1, 33 (22%) for HHV5, 6 (4%) for HHV4 and 3 (2%) for HHV6, respectively. Moreover, double-infection was found in 22 out of 59 specimens (37%), most of which were double-infection of HHV1 and HHV5s. However, relationship between virus infection and sperm abnormality was not identified.[br]This study demonstrated a high-frequency of viral infection in human male infertile patients. However, we did not observe significant difference in abnormality in semen volume, sperm number and sperm mortality between virus-infected and -uninfected patients. The results may indicate that viral infection in human has no relation to male infertility but we did not examine the expression of viral gene, especially viral TK gene. Accordingly, it is indispensable to examine whether viral TK gene is expressed in the testis of virus-infected infertile patient.[br][br]Nothing to Disclose: MC, MH, L-YC, S-iI, TK, YK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1465 51 195 SAT-74 PO35-02 Saturday 134 2012


135 ENDO12L_SAT-75 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) The Putative Humanin Receptor Subunits Are Present in the Testis Prasanth Surampudi, Yue Jia, YanHe Lue, Jeffrey Kim, Christina Wang, Ronald Swerdloff Labiomed at Harbor UCLA, Torrance, CA [bold]Background:[/bold] Humanin (HN), an 24 amino acid mitochondrial peptide, is found in several tissues including the brain and testis. Our previous studies demonstrated that administration of HN peptide protects testicular germ cell from apoptosis induced by hormonal deprivation in vivo [amp] hyperthermia in vitro. It has been demonstrated in neuronal cells that the cytoprotective action of HN is mediated in part through a membrane bound trimeric receptor, comprised of subunits WSX-1, GP-130, and CNTFR[alpha], that utilizes STAT3 as a signaling pathway. Thus, we hypothesize that the cytoprotective action of HN may be mediated through a similar receptor in the testis.[br][bold]Objective:[/bold] We attempted to identify and localize the presence of the HN receptor subunits WSX1, GP130, and CNTFR[alpha] in mouse testes.[br]Methods: Adult (12-week-old) mice (C57BL/6J, control and GnRH antagonist treated for two weeks) were used in the study. Immediately after sacrifice, one testis from each animal was dissected and snap-frozen in liquid nitrogen for protein extraction and Western blot (WB). The contralateral testis was perfused with Bouin[apos]s solution. Paraffin embedded testicular sections were subjected to immunohistochemistry (IHC) and immunofluorescence to determine the localization of the putative HN receptor subunits.[br][bold]Results:[/bold] Western blot showed that WSX1 was detected in the testes as at [sim]70kDa, which was absent in spleen samples from WSX1 knockout mice. GP130 was detected as a 130 kDa protein in the testes. CNTFR[alpha] was not detected in testes by WB using the available antibodies studied previously in neuronal cells. IHC revealed the presence of strong immunoreactivity of WSX1, GP130 [amp] CNTFR-[alpha] in Leydig cells. We found that WSX1 [amp] GP130 were localized in the cytoplasm of spermatocytes, but not CNTFR[alpha]. There were strong immunoexpression of both WSX1 [amp] GP130 in spontaneous apoptotic germ cells in control mice, and in apoptotic germ cells induced by testicular hormonal deprivation in GnRH-antagonist treated mice.[br][bold]Conclusion[/bold]: 1) HN receptor subunits WSX1, GP130 and CNTF[alpha] are present in mouse testes; 2) These three subunits of HN receptor are localized in Leydig cells; 3) Both WSX1 and GP130 are present in the cytoplasm of spermatocytes and apoptotic germ cells. Functional studies are ongoing in our laboratory to determine the role of each receptor subunit. This will help us understand the mechanism of the cytoprotective actions of HN in the testes and its modulation for clinical applications.[br][br]Disclosures: RS: Investigator, Aeterna-Zentaris. Nothing to Disclose: PS, YJ, YL, JK, CW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2274 51 196 SAT-75 PO35-02 Saturday 135 2012


136 ENDO12L_SAT-76 POSTER SESSION: Male Reproduction [amp] Fertility (1:30 PM-3:30 PM) Humanin Is Detected in the Testis after Intraperitoneal (ip) Administration Yue Jia, Junxiang Wan, Yanhe Lue, Aikoui Ohanyan, Kuk Wha Lee, Ronald S Swerdloff, Pinchas Cohen, Christina CL Wang Los Angeles Biomedical Research Institute and Harbor-UCLA Medical Center, Torrance, CA; Mattel Children[apos]s Hospital, David Geffen School of Medicine at UCLA, Los Angeles, CA Objective: Humanin (HN) has been identified as an endogenous cytoprotective peptide. HN protects neuronal, pancreatic beta, and cardiac cells from apoptosis after stress. In our previous study, intra-testicular injection of HN protected male germ cells from apoptosis after hormonal deprivation. The present study determines the distribution of HN in the testis after ip injection of HN.[br]Methods: In experiment 1, adult male mice (C57BL/6, 14 weeks old) received a single ip dose of HN peptide (40 mg/Kg body weight) and sacrificed at 15, 30, 60, 120, and 240 minutes after injection. In experiment 2, male mice were sacrificed at 240 minutes after different ip doses (4, 10, and 40mg/Kg/BW) HN peptide. Control mice only received placebo (saline) injection. One testis of each animal was collected and kept at -80C for Western blot and a specific ELISA assay used to detect the level of exogenous HN. The other testis was fixed with Bouin[apos]s fixative for immunohistochemistry to localize the HN peptide.[br]Results: In experiment 1, two peaks were detected by ELISA after 40 mg/Kg/BW HN ip injection, one at 15 minutes (6.9 ng/mg)(vascular phase) and the second at 240 minutes (8.0 ng/mg) (tissue phase). Western blot analyses confirmed that exogenous HN was detected at 15 and 240 minutes. In experiment 2, HN level in testis was 1.6 ng/mg in the control mice, rose after HN 4 and 10 mg/Kg/BW, reaching 8.6 ng/mg after 40 mg/Kg/BW HN ip injection. Exogenous HN expression was only detected after HN 40mg/Kg injection by immuno-blot. Using immunohistochemistry, specific HN staining was found in Leydig and early germ cells in mouse testis at 240 minutes after 40mg/Kg/BW HN injection.[br]Summary: Four hours after ip injection of 40 mg/Kg/BW HN peptide, exogenous HN peptide was detected in testis and localized mainly in Leydig cells and early germ cells. These data help define target sites for exogenous HN in the testes and may have clinical application since preliminary data from our laboratory shows that ip injection of HN at 40mg/Kg/BW prevents cyclophosphamide induced acute pro-apoptotic effect on male germ cell in mice.[br][br]Disclosures: RSS: Investigator, Aeterna-Zentaris. Nothing to Disclose: YJ, JW, YL, AO, KWL, PC, CCLW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1251 51 197 SAT-76 PO35-02 Saturday 136 2012


137 ENDO12L_SAT-85 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Retinaldehyde Dehydrogenase 1 Is a Novel Regulator of Adaptive Thermogenesis in White Adipose Tissue Florian W Kiefer, Cecile Vernochet, Patrick O[apos]Brian, Steffen Spoerl, Jonathan D Brown, C Ronald Kahn, Jorge Plutzky Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA; Harvard Medical School, Boston, MA Two functionally distinct types of fat are present in mammals: white adipose tissue (WAT), the primary site of triglyceride storage, and brown adipose tissue (BAT), which promotes energy dissipation through adaptive thermogenesis. Factors that determine white versus brown fat differentiation and function remain poorly understood. Recent data link vitamin A and its retinoid metabolites to the regulation of adipogenesis and energy balance. Retinoid metabolism is tightly controlled by an enzymatic network in which retinaldehyde dehydrogenases (Aldhs) are the rate-limiting enzymes converting retinaldehyde (Rald) to retinoic acid. Previously, we demonstrated that lack of the Aldh isoform 1a1 protected mice from diet-induced obesity by inducing a hypermetabolic state. However, the mechanisms for this phenotype remained unclear.[br]Here we show that Aldh1a1 is predominately expressed in white but not brown fat. Genetic Aldh1a1 deficiency resulted in increased expression of classic BAT markers in WAT of standard chow-fed mice. Moreover, Aldh1a1-deficient mice manifested increased mitochondrial enzyme activity and oxygen consumption in WAT and were resistant to cold exposure as compared to controls. Using antisense approaches, WAT-selective Aldh1a1 knockdown limited weight gain and improved glucose homeostasis in high fat-fed obese mice by induction of a similar thermogenic program. In investigating underlying mechanisms, we found that Rald, whose endogenous concentrations are elevated in Aldh1a1 deficiency, is a transcriptional regulator of uncoupling protein 1 (UCP1), the key mediator of adaptive thermogenesis. Rald stimulation in white adipocytes increased UCP1 expression 100-fold in a retinoic acid receptor (RAR) but not retinoic X receptor (RXR)-dependent manner. Rald selectively bound and activated RAR but not RXR, induced UCP1 promoter activity, and recruited the co-activator PGC-1[alpha] to the UCP1 promoter.[br]These data establish Aldh1a1 and Rald as novel determinants of adipocyte plasticity and adaptive thermogenesis in white adipose tissue with potential therapeutic consequences for obesity and type 2 diabetes.[br][br]Sources of Research Support: HL048743, AR054604-03S1, NIH/NIDDK DK056626, DK048873, DK048873-14S2, and the Austrian Science Fund (FWF): J3107-B19.[br][br]Nothing to Disclose: FWK, CV, PO, SS, JDB, CRK, JP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 225 52 198 SAT-85 PO27-02 Saturday 137 2012


138 ENDO12L_SAT-86 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Adipose Tissue[ndash]Specific Deletion of the Mitochondrial Transcription Factor A (TFAM) Causes Lipodystrophy and Hepatosteatosis Cecile Vernochet, Ron C Kahn Joslin Diabetes Center, Boston, MA Mitochondrial dysfunction in adipose tissue has been reported in obesity, type 2 diabetes and metabolic syndrome, but whether this dysfunction participates in the development or is a result of these disorders remains an open question. Mitochondrial transcription factor A (Tfam) plays a key role in mitochondrial function by controlling both mtDNA stability and transcription. To investigate the physiological consequences of mitochondrial impairment in adipose tissue, we generated a mouse deficient for Tfam specifically in adipocytes (Ad-TFKO) using Adiponectin-Cre and Tfam floxed allele. Ad-TFKO mice are leaner than control mice on normal chow diet (CD) by 16% and 28% less after 6 weeks upon HFD. In the CD, Ad-TFKO mice brown adipose tissue was almost inexistent and completely overtaken by white adipose tissue, whereas subcutaneous white fat was reduced by 61% and perigonadal fat by 52%. Ad-TFKO mice also had 14% decreased in basal energy expenditure with no significant change in spontaneous activity. Interestingly, fed glucose levels in Ad-TFKO mice were elevated by 35% compared to controls but fasting glucoses were similar. Ad-TFKO also exhibited a similar glucose clearance during an insulin tolerance test but remained hyperglycemic compared to control throughout. Ad-TFKO has a 2 fold increase in liver mass and histology revealed marked triglycerides accumulation. Thus TFAM KO in fat impaired white and brown adipose tissue expansion and provoked lipid localization to the liver leading to hepatosteatosis and systemic insulin resistance. These results thus suggest that mitochondria dysfunction in adipose tissue could indeed participate and accelerate development of metabolic syndrome.[br][br]Nothing to Disclose: CV, RCK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1550 52 199 SAT-86 PO27-02 Saturday 138 2012


139 ENDO12L_SAT-87 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Unacylated Ghrelin Suppresses High-Fat Diet[ndash]Induced Phenotypic Switch in Adipose Tissue Macrophages, and Enhances Differentiation of Bone Marrow-Derived Endothelial Progenitor Cells Patric JD Delhanty, Lucy Baldeon-Rojas, Martin Huisman, Iris van den Berge, Thierry Abribat, Axel PN Themmen, Pieter JM Leenen, AJ van der Lely Erasmus Medical Center, Rotterdam, Netherlands; Erasmus Medical Center, Rotterdam, Netherlands; Metabolic Diseases, Lyon, France The incidence of obesity and type 2 diabetes (T2D) is of epidemic proportions, although pre-diabetes, characterized by insulin-resistance (IR) occurs in an even larger population. The development of T2D, and associated morbidities, would likely be prevented by reversal of insulin-resistance. We now have evidence that unacylated ghrelin (UAG) has this beneficial effect, although the mechanisms are unclear. Obesity-induced conversion of anti-inflammatory macrophages (MP) to an inflammatory phenotype in fat is an important effector of IR, and is thought to be driven by recruitment of inflammatory monocytes from bone marrow (BM). To investigate if UAG modulates these processes, we infused mice for 4 wks with either saline or UAG. Half these mice were given a normal diet (ND), and half a high fat diet (HFD) during the last 2 wks of infusion to induce IR and increased fat mass. Epididymal fat was removed at wk 4, and the stromal-vascular cell fraction (SVF) isolated and analyzed by FACS. The MP population in the SVF was defined using F4/80 as a marker. Inflammatory (M1) and anti-inflammatory (M2) adipose tissue MPs (ATM) were defined by CD11c or CD301 immunoreactivity, respectively. Compared with the ND, HFD caused a significant 5-fold increase in ATM, accompanied by significant induction (from 6[plusmn]1 to 13[plusmn]0.8%, p[lt]0.001) of M1, and suppression (97.4[plusmn]0.3 to 96.2[plusmn]0.4%, p[lt]0.01) of M2 cells in the ATM population. However, UAG treatment significantly reversed these effects (M1, 6.1[plusmn]1%; M2, 98.8[plusmn]0.2% of ATM). To investigate if this was related to modulation of BM function, we assessed monocyte populations in BM of [plusmn]HFD/[plusmn]UAG treated mice, but surprisingly found no effect of either HFD or UAG. To explore this further, we stimulated BM cells in vitro to develop into three lineages: endothelial progenitor cells (EPC), MPs and dendritic cells (DC). No effect of HFD was observed on recovery of these cell types, and UAG did not affect the differentiation of MPs or DC. However, UAG stimulated the differentiation of EPC 126[plusmn]9% (p[lt]0.05) compared to controls. These data suggest short-term HFD causes an M2 to M1 transition in ATM that is corrected by UAG treatment. Thus, UAG appears to have a direct effect on fat-resident MP phenotype, but not on BM composition. UAG agonists may prevent obesity-induced IR in part by blocking its pro-inflammatory effects in fat. UAG also promoted the potential of BM to generate EPC, with potentially cardiovascular protective properties.[br][br]Nothing to Disclose: PJDD, LB-R, MH, IvdB, TA, APNT, PJML, AJvdL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 921 52 200 SAT-87 PO27-02 Saturday 139 2012


140 ENDO12L_SAT-88 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Estrogen and Retinoid Interactions in the Regulation of Adipose Triglyceride Lipase Barbara Reichert, Rumana Yasmeen, Fangping Yang, Petra Kotzbeck, Margret Poeschl, Rudolf Zechner, Ouliana Ziouzenkova Ohio State University, Columbus, OH; Ohio State University, Columbus, OH; University of Graz, Graz, Austria Visceral obesity is a complex disorder involving dysregulation of fat storage and lipolysis. Identifying mechanisms and regulation of the rate-limiting enzyme of lipolysis in adipose tissue, adipose triglyceride lipase (ATGL), is key in understanding and treating this disorder. Our previous studies implicated vitamin A metabolites, retinaldehyde and retinoic acid (RA), as key regulators of visceral fat accumulation in females. Here, we investigate the effects of estrogen and vitamin A metabolites in regulating ATGL.[br]We utilized ovariectomized mice as a model of postmenopause to examine the roles of vitamin A and estrogen in vivo. We ovariectomized (OVX) or sham-operated wild-type (WT; n=9 OVX, n=19 sham) and Aldh1a1-/- (n=10 OVX, n=10 sham) mice, which lack the principal enzyme for RA production from retinaldehyde in adipose tissue. Tissues were collected three months after surgery from mice receiving a regular chow diet. Ovariectomized WT mice acquired 240% more visceral fat than WT sham. Ovariectomized mouse visceral fat had increased Aldh1a3 (244%) and RA-sensitive Cyp26A1 expression compared to sham, suggesting increased RA production in estrogen-deprived mice. Ovariectomized Aldh1a1-/- mice with disrupted RA production resisted visceral fat accumulation and had improved glucose tolerance. Correspondently, ATGL protein in visceral fat was 190% higher in Aldh1a1-/- compared to WT ovariectomized mice.[br]To investigate the influences of estrogen and vitamin A seen in vivo, we utilized mature 3T3-L1 adipocytes and human SGBS adipocytes in vitro. We stimulated these adipocytes with retinaldehyde or RA in the presence and absence of estrogen. ATGL protein levels were measured by Western blot and lipase activity by non-esterified fatty acid (NEFA)-release assay. 3T3-L1 adipocytes stimulated with retinaldehyde had increased ATGL protein. Retinaldehyde or estrogen increased NEFA release in SGBS adipocytes; however combination of these metabolites did not further increase NEFA release, suggesting a common pathway for retinaldehyde and estrogen action.[br]In conclusion, our data suggest that estrogen suppresses retinaldehyde catabolism by Aldh1, and retinaldehyde, in turn, induces ATGL and protects ovariectomized Aldh1a1-/- female mice from visceral obesity. By elucidating Aldh1[apos]s role in sexually dimorphic fat accumulation, we can potentially target visceral obesity in women.[br][br]Sources of Research Support: Alpha Omega Alpha Carolyn L. Kuckein Student Research Fellowship.[br][br]Nothing to Disclose: BR, RY, FY, PK, MP, RZ, OZ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1546 52 201 SAT-88 PO27-02 Saturday 140 2012


141 ENDO12L_SAT-89 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) FGF21 Suppresses Lipolysis Via Akt-Dependent Reduction of cAMP Levels in Mouse and Human Adipose Tissues Xuan Ge, Aimin Xu, Cheng Chen, Karen SL Lam The University of Hong Kong, Hong Kong, China; The University of Hong Kong, Hong Kong, China Fibroblast Growth Factor 21 (FGF21) is a liver-secreted hormone that plays a regulatory role in glucose and lipid metabolism in animals. During fasting, free fatty acids (FFA) released from the adipose tissue stimulate hepatic FGF21 production [italic]via[/italic] the activation of PPAR[alpha]. However, the role of FGF21 in lipolysis and the underlying mechanism remain poorly characterized. This study aimed to address these questions using in vivo and ex vivo approaches as well as the FGF21 knockout (KO) mice. A single bolus injection of FGF21 acutely reduced basal and isoproterenol-stimulated serum FFA and glycerol levels in C57 mice and FGF21 KO mice. A greater suppression of lipolysis was seen in the FGF21 KO mice which also had higher baseline serum FFA and glycerol levels. At the molecular level, such an inhibitory effect of FGF21 on lipolysis was accompanied by the decreased phosphorylation of hormone sensitive lipase (HSL) at ser660. Explant studies showed that FGF-21 induced Akt phosphorylation in both mouse and human white adipose tissue (WAT). This was accompanied by an elevation in phosphodiesterase 3B (PDE3B) activity, a reduction in foskolin (FSK)[ndash]stimulated cAMP level, and decreased basal and FSK-stimulated lipolysis, all of which could be attenuated by an Akt inhibitor. Taken together, these findings suggest that FGF21 plays a physiological role in regulating basal lipolysis activity, and suppresses basal and stimulated lipolysis through a reduction in cAMP levels, consequent to Akt-dependent activation of PDE3B, in both mouse and human WAT.[br][br]Sources of Research Support: Acknowledgement: This study was supported by a Collaborative Research Grant (HKU3/CRF/09) from the Hong Kong Research Grant Council.[br][br]Nothing to Disclose: XG, AX, CC, KSLL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 102 52 202 SAT-89 PO27-02 Saturday 141 2012


142 ENDO12L_SAT-90 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) SRA/SRAP Regulates Adipocyte Differentiation and Insulin Sensitivity through Insulin Receptor Signaling Shannon Liu, Ruichuan Xu, Isabelle Gerin, William P Cawthorn, Ormond A MacDougald, Alan R Saltiel, Ronald J Koenig, Bin Xu University of Michigan Medical Center, Ann Arbor, MI; University of Michigan Medical Center, Ann Arbor, MI; Universit[eacute] Catholique de Louvain, Brussels, Belgium; University of Michigan, Ann Arbor, MI; University of Michigan Medical Center, Ann Arbor, MI The Sra1 gene expresses both a non-coding RNA denoted Steroid Receptor RNA Activator (SRA), and an encoded protein (SRAP). Our previous work showed that SRA enhances adipogenesis and insulin sensitivity. To assess the mechanism, we differentiated the ST2 mesenchymal precursor cell line into adipocytes using either a standard hormone cocktail of methylisobutylxanthine, dexamethasone and insulin, or all possible 1 and 2 hormone combinations. These studies showed that SRA overexpression promotes ST2 cell differentiation to adipocytes if the hormone cocktail includes insulin, suggesting that the SRA effect involves insulin signaling. In addition, SRA overexpression inhibits phosphorylation of p38 mitogen activated protein kinase (MAPK) and c-Jun NH2-terminal kinase (JNK) in the early mitotic clonal expansion of ST2 cells. Conversely, knockdown of endogenous SRA/SRAP increases phosphorylation of JNK. Next, we used 3T3-L1 cells to study the effects of SRA/SRAP on glucose uptake. Overexpression of SRAP in 3T3-L1 adipocytes increased insulin-stimulated glucose uptake. Knockdown of endogenous SRA/SRAP in 3T3-L1 mature adipocytes increased basal glucose uptake [sim]2 fold, whereas it inhibited glucose uptake in response to high doses of insulin. IGF-I receptor and GLUT4 proteins were induced by SRA/SRAP knockdown, and may contribute to the associated increased basal glucose uptake. Importantly, knockdown of SRA/SRAP in mature 3T3-L1 adipocytes leads to reduce protein levels of both insulin receptor [beta] (IR[beta]) and its precursor, without changing insulin receptor mRNA levels. This effect is partially eliminated by the proteasome inhibitor MG132, suggesting that SRA/SRAP negatively regulates proteasomal degradation of IR[beta] and its precursor. SRA/SRA knockdown also results in decreased autophosphorylation of IR[beta] and tyrosine phosphorylation of insulin receptor substrate -1 (IRS-1), but these changes are only partially accounted for by decreased IR[beta] protein. We found that SRA/SRAP directly binds to the insulin receptor, suggesting the effects on IR[beta] and its precursor may result from physical interaction. In summary, the present study reveals that SRA/SRAP regulates adipocyte differentiation and insulin sensitivity through maintaining the protein content of the insulin receptor and modulating insulin receptor signaling.[br][br]Nothing to Disclose: SL, RX, IG, WPC, OAM, ARS, RJK, BX 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1570 52 203 SAT-90 PO27-02 Saturday 142 2012


143 ENDO12L_SAT-91 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) The Clock Gene Bmal1 Regulates Adipogenesis Via Wnt Signaling Pathway Bingyan Guo, Somik Chatterjee, Lifei Li, Ji M Kim, Jeongkyung Lee, Vijay K Yechoor, Laurie J Minze, Willa A Hsueh, Ke Ma The Methodist Hospital Research Institute, Houston, TX; Baylor College of Medicine, Houston, TX Circadian clock is present in adipose tissue, but whether it regulates adipocyte development is unknown and the molecular link between circadian dysregulation and obesity is poorly understood. Here we report that disruption of the clock gene, brain and muscle Arnt-like 1(Bmal1), in mice led to increased adipogenesis, adipocyte hypertrophy and obesity, which was evident as early as day 7 of postnatal development without significant alterations in overall energy homeostasis, compared to wild-type controls. This is due to its cell-autonomous effect as Bmal1 deficiency from embryonic fibroblasts, as well as stable shRNA knockdown (KD) in 3T3-L1 preadipocyte and C3H10T1/2 mesenchymal stem cells promoted adipogenic differentiation. We demonstrate that attenuation of Bmal1 function resulted in down-regulation of many genes in Wnt signaling pathway, a known inhibitory mechanism of adipogenesis. Analysis of promoters of these genes ([beta]-catenin, Dishevelled2, TCF3) revealed Bmal1 occupancy, indicating direct circadian regulation by Bmal1, and serum shock was able to elicit their intrinsic circadian oscillation patterns. As a result, Bmal1 KD suppressed Wnt signaling activity while its overexpression augmented this activity. In addition, Wnt3a treatment partially restored the blunted Wnt activity, and in turn, suppressed the increased adipogenesis in KD cells. Taken together, our study demonstrates a novel function of the core clock gene Bmal1 as a negative regulator of adipocyte development and this function is mediated through direct transcriptional regulation of canonical Wnt signaling pathway. Our study suggests that this molecular pathway could be the mechanistic link between circadian disruption and development of obesity.[br][br]Nothing to Disclose: BG, SC, LL, JMK, JL, VKY, LJM, WAH, KM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1950 52 204 SAT-91 PO27-02 Saturday 143 2012


144 ENDO12L_SAT-92 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Testosterone Inhibits Adipogenesis through Modulation of the BMP4 Signaling Pathway Gregorio Chazenbalk, Prapti Singh, David Abott, Dumesic Daniel David Geffen School of Medicine at UCLA, Los Angeles, CA; University of Wisconsin, Madison, WI OBJECTIVE: Adipogenesis is a paracrine-mediated process whereby adipose stem cells (ASCs) become preadipocytes through commitment and then differentiate to adipocytes. Androgen acts through its receptor (AR) to inhibit human subcutaneous (SC) abdominal preadipocyte differentiation. We have previously demonstrated that testosterone (T) inhibits adipogenesis during conversion of ASCs to preadipocytes. The present study utilizes SC abdominal adipose from nonobese women to determine if the inhibitory effect of T in adipogenesis is exclusively mediated through AR events and if occurs during ASC commitment to preadipocytes.[br]DESIGN: Prospective study[br]MATERIAL AND METHODS: Lipoaspirate was obtained from SC abdominal adipose of nonobese patients. ASCs were isolated and then incubated in adipogenic medium for 3 days without and with testosterone (T, 50 nM), 5[alpha]-Dihydrotestosterone (DHT, 5 nM), androgen receptor (AR) antagonist flutamide (F, 100 nM), and Bone Morphogenetic Protein 4 (BMP4, 5 nM), a major regulator of ASC commitment to preadipocytes. Expression of adipogenic gene markers PPARg, CEBP-[alpha] and CEBP-[beta] was determined by qRT-PCR and expressed as fold changes (treated vs. untreated samples).[br]RESULTS: T decreased gene expression of PPARg (0.5 fold, p[lt]0.001), CEBP-[alpha] (0.6 fold, p[lt]0.001) and CEBP-[beta] (0.7 fold, p[lt]0.001). Similarly, DHT decreased gene expression of PPARg (0.4 fold, p[lt]0.002), CEBP-[alpha] (0.5 fold, p[lt]0.003) and CEBP-[beta] (0.7 fold, p[lt]0.001). F partially reversed T inhibition of PPAR[gamma] (0.3 fold, p[lt]0.02 T vs T+F, p[lt] 0.006 F vs F+T), CEBP-[alpha] (0.3 fold, p[lt]0.007 T vs T+F, p[lt] 0.003 F vs T +F) but completely reversed the inhibitory effect of T in CEBP-[beta] (0.3 fold, p[lt] 0.008 T vs T+F, n.s. F vs T+F). BMP4 stimulated PPAR[gamma] (1.96 fold, p[lt]0.010) and CEBP-[alpha] (1.88 fold, p[lt]0.001) and these BMP4 actions were inhibited by T. BMP4 did not alter CEBP-[beta] gene expression.[br]SUMMARY: Androgen inhibits early stages of adipogenesis through the BMP4 signaling pathway and this action appears mediated in part through aromatization of androgen to estrogen.[br]CONCLUSIONS: Androgen excess may diminish SC abdominal adipogenesis and reduce the capacity of this adipose store to safely store fat.[br][br]Nothing to Disclose: GC, PS, DA, DD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2292 52 205 SAT-92 PO27-02 Saturday 144 2012


145 ENDO12L_SAT-93 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Do Differences in Metabolic Function between Adipose Tissue Depots Play a Role in Obesity Development? Korinna Huber, Mirijam Bedurftig, Jennifer Miles-Chan, Nichola Thompson, Bernhard H Breier University of Veterinary Medicine, Hannover, Germany; University of Fribourg, Fribourg, Switzerland; The University of Adelaide, Adelaide, Australia; Massey University, Albany Campus, Auckland, New Zealand Successful prevention and therapeutic strategies to combat the growing [ldquo]obesity crisis[rdquo] are hampered by our lack of understanding of different pathways to obesity. A number of metabolically distinct pathways to obesity with different health outcomes are recognised (1,2,3). Here we investigate the role of adipose tissue and underlying mechanisms of one such pathway through interrogation of responses to metabolically active hormones [italic] ex vivo [/italic] and the expression of molecular markers of insulin signalling and lipolytic responses in both subcutaneous (ScAT) and retroperitoneal adipose tissue (RpAT).[br]Pregnant Wistar rats were either fed chow [italic] ad libitum [/italic] or undernourished throughout pregnancy, generating control (AD) or prenatally undernourished (UN) offspring (1). At 9 months of age, ScAT and RpAT were collected to investigate insulin-stimulated glucose uptake, catecholamine-induced glycerol release and insulin signalling proteins.[br]The mean adipocyte size was significantly higher (p[lt]0.001) in both types of adipose tissues from UN in comparison with AD offspring. Insulin increased glucose uptake [italic] ex vivo [/italic] in ScAT from both AD and UN offspring (p[lt]0.001); the response to insulin was stronger in AD offspring (p[lt]0.01). In RpAT, insulin stimulated glucose uptake only in AD offspring (p[lt]0.001) but not in UN offspring. While protein expression of insulin signalling markers, InsR, PI3K, PKC[zeta] and mTOR, were similar in ScAT of the two groups, significantly lower levels (p[lt]0.01) were observed in RpAT of UN in comparison with AD offspring. Lipolytic responses to [beta]3-adrenergic stimulation [italic] ex vivo [/italic] differed between adipose tissue depots and were influenced by prenatal nutrition. Lipolytic responses in ScAT were present (p[lt]0.01) in UN offspring, but were significantly lower (p[lt]0.01) compared to AD offspring. The lower lipolytic response in ScAT from UN offspring was completely inhibited by insulin, while in AD offspring, insulin only partially reduced (p[lt]0.01) the stimulation of glycerol release. In contrast, [beta]3-adrenergic stimulation of lipolysis was similar in RpAT from AD and UN offspring and insulin-dependent inhibition of glycerol release was absent.[br]These findings emphasize the role of different adipose tissue depots in the pathophysiology of obesity. A shift in insulin sensitivity and lipolytic responses in different adipose tissue depots may explain specific patterns of fat storage and energy availability that contribute to distinct health consequences.[br][br](1) Thompson NM, Norman AM, Donkin SS, Shankar RR, Vickers MH, Miles JL, Breier BH (2007). Prenatal and postnatal pathways to obesity: different underlying mechanisms, different metabolic outcomes. Endocrinology 148(5): 2345-2354. (2) Huber K, Miles JL, Norman AM, Thompson NM, Davison M, Breier BH (2009). Prenatally-induced changes in muscle structure and metabolic function facilitate exercise-induced obesity prevention. Endocrinology. 150 (9): 4135-4144. (3) Miles JL, Huber K, Thompson NM, Davison M and Breier BH (2009). Moderate daily exercise activates metabolic flexibility to prevent prenatally-induced obesity. Endocrinology. 150 (1):179-186.[br][br]Nothing to Disclose: KH, MB, JM-C, NT, BHB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 841 52 206 SAT-93 PO27-02 Saturday 145 2012


146 ENDO12L_SAT-94 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Nutrition-Independent Morphological Changes in Adipose Tissue during Critical Illness Mirna Bastos Marques, Lies Langouche, Annelies Aertgeerts, Sarah Vander Perre, Sarah Derde, Greet Van den Berghe KU Leuven, Leuven, Belgium We previously reported that adipose tissue biopsies from critically ill patients display an increased number of small adipocytes and a profound infiltration by M[sub]2 [/sub]type macrophages (1,2). Infiltration of M[sub]1 [/sub]type macrophages in adipose tissue has been described in conditions of excessive food intake. It currently remains unknown whether the morphological changes in adipose tissue during critical illness are related to the administration of parenteral nutrition. This study was designed to compare the impact of early parenteral nutrition (PN) with fasting on the morphological alterations within adipose tissue evoked by critical illness.[br]Male mice of 24-weeks were anesthetized and instrumented with central venous catheters. Sepsis was produced by cecal ligation and puncture. Mice were fluid resuscitated and treated with antibiotics. After 24-hour fluid resuscitation, mice were randomly allocated to fasting (n=11) or feeding (PN) (n=11). For comparison, healthy mice (n= 11) were allowed [italic]ad libitum[/italic] chow. Total fat content of the body was quantified with DEXA on days 0 and 5. On day 5, mice were sacrificed and adipose tissue was collected. Adipocyte size was determined in HE-stained sections through computer imaging analysis. Macrophages were identified by ICC (F4/80 Ab) and scored for intensity. Macrophage markers TNF[alpha] (M[sub]1[/sub]) and arginase (M[sub]2[/sub])[sub] [/sub]were quantified with real time PCR.[br]In subcutaneous and visceral adipose tissue from critically ill animals, adipocytes werer significantly smaller than in healthy controls. This change in adipocyte size with illness was unaffected by nutrition. Total fat content was lower in ill than in healthy animals, but again irrespective of nutrition. More macrophages were present in adipose tissue from ill than from healthy mice and, again, this was irrespective of nutrition. The increased number of macrophages in adipose tissue were predominantly of the M[sub]2[/sub] type, as arginase levels were 32-fold increased in all adipose depots, and TNF[alpha] was only slightly elevated.[br]In conclusion, the appearance of small adipocytes infiltrated with more M[sub]2 [/sub]type macrophages during critical illness was independent of PN administration. More small adipocytes during critical illness may reflect an adaptive response to illness, taken their increased capacity to take up glucose and fatty acids. In addition, infiltration of adipose tissue by M[sub]2[/sub] macrophages may be adaptive taken their antiinflammatory and insulin-sensitizing feature. This remains to be investigated.[br][br](1) Langouche L et al, Am J Respir Crit Care Med 2010, 182(4): 507-516. (2) Langouche et al, Crit Care. 2011;15(5):R245.[br][br]Nothing to Disclose: MBM, LL, AA, SVP, SD, GVdB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1431 52 207 SAT-94 PO27-02 Saturday 146 2012


147 ENDO12L_SAT-95 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Increased Mitochondrial Activity in BMP7-Treated Brown Adipocytes, Due to Increased CPT1- and CD36-Mediated Fatty Acid Uptake Kristy L Townsend, Ding An, TianLian Huang, Matthew D Lynes, Hongbin Zhang, Laurie J Goodyear, Yu-Hua Tseng Joslin Diabetes Center, Harvard Medical School, Boston, MA; Boston University, Boston, MA Brown adipose tissue dissipates chemical energy in the form of heat and has recently been shown to actively uptake triglyceride in a regulated manner. Factors that directly control fatty acid uptake and oxidation in brown adipocytes have not yet been fully elucidated. Bone morphogenetic protein 7 (BMP7) is a growth factor capable of inducing brown fat mitochondrial biogenesis during differentiation from adipose progenitors. Administration of BMP7 to mice also results in increased energy expenditure. To determine if BMP7 is able to directly affect the mitochondrial activity of mature brown adipocytes, independent of the differentiation process, we delivered BMP7 to mature brown adipocytes and measured mitochondrial activity. We found that BMP7 increased mitochondrial activity, including fatty acid oxidation and citrate synthase activity. This was accompanied by an increase in fatty acid uptake, and increased protein expression of CPT1 and CD36, which import fatty acids into the mitochondria and the cell, respectively. Importantly, inhibition of either CPT1 or CD36 resulted in a blunting of the mitochondrial activity of BMP7-treated cells. In conclusion, BMP7 increases mitochondrial activity in mature brown adipocytes via increased fatty acid uptake and oxidation, a process which requires the fatty acid transporters CPT1 and CD36.[br][br]Sources of Research Support: K.L.T. was supported by NIH T32-DK007260[ndash]33 and NIH F32DK091996, This work was supported in part by NIH grants DK077097 (Y.-H.T.), and Joslin Diabetes Center[apos]s Diabetes Research Center (P30 DK036836 from the NIDDK), a research grant from the Eli Lilly Research Foundation and by funding from Harvard Stem Cell Institute (to Y.-H.T.).[br][br]Nothing to Disclose: KLT, DA, TH, MDL, HZ, LJG, Y-HT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 577 52 208 SAT-95 PO27-02 Saturday 147 2012


148 ENDO12L_SAT-96 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Total and High-Molecular-Weight Adiponectin Levels Throughout Life in bGH and GHR-/- Mice Ellen R Lubbers, Aubree K Ziegler, Edward O List, John J Kopchick, Darlene E Berryman Ohio University, Athens, OH; Ohio University, Athens, OH; Ohio University, Athens, OH Bovine growth hormone (bGH) transgenic mice are giant and lean, while growth hormone receptor knockout (GHR-/-) mice are dwarf and obese. The bGH mice have significantly reduced lifespan and reduced insulin sensitivity. In contrast, GHR-/- mice have increased longevity and improved insulin sensitivity. Adiponectin is a powerful insulin sensitizer. It has been suggested that the high molecular weight (HMW) form has the predominate insulin sensitizing bioactivity. The levels of total and HMW adiponectin have been presented in these mice at 6 months of age, with total adiponectin being decreased in bGH mice, but increased in GHR-/-mice compared to wild type (WT) controls. HMW adiponectin and the HMW/total adiponectin ratio was also shown to be increased in GHR-/- mice, but not changed in bGH(1). As adiposity and insulin sensitivity change with age in these mice and a single 6-month time point represents different relative ages in relation to the total lifespan of the mice, an assessment of total and HMW adiponectin throughout life using several time points could help elucidate the role of adiponectin in these unique mouse lines. Serum total and HMW adiponectin were measured side-by-side using an ALPCO ELISA kit. In the bGH mice, total adiponectin is reduced at 2, 6 and 9 months of age, but is not significantly changed at 14 months of age; HMW adiponectin is significantly reduced only at 2 months of age. In bGH mice, the HMW/Total adiponectin ratio showed no significant difference from WT at any age. Circulating total and HMW adiponectin in GHR-/- mice are increased throughout life, at 6, 12 and 24 months of age. Interestingly, the HMW/total adiponectin ratio is increased in GHR-/- mice only at 6-months of age. While observed serum adiponectin levels remain relatively consistent, genotype differences become less pronounced with advancing age. Serum adiponectin will also be compared to leptin and insulin levels in the same samples. Overall, these changes in adiponectin during lifespan highlight the importance of carefully selecting time points for sample collection, as differences of a few months can alter observed genotype differences.[br][br](1)E Lubbers, BMiles, EO List1, JJ Kopchick, DE Berryman. Total and high molecular weight adiponectin levels in mice with altered GH signaling. Poster presented at: Experimental Biology; 2010 April 24-28; Anaheim, CA.[br][br]Sources of Research Support: This work was supported in part by the State of Ohio[apos]s Eminent Scholar Program that includes a gift from Milton and Lawrence Goll, by the AMVETS, by the Diabetes Research Initiative at Ohio University, by NIH grants DK083729, AG031736, and by a Student Enhancement Award.[br][br]Nothing to Disclose: ERL, AKZ, EOL, JJK, DEB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1773 52 209 SAT-96 PO27-02 Saturday 148 2012


149 ENDO12L_SAT-97 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Quantification of Collagen and Effect on Adipocyte Size in bGH Mice Katie M Troike, Lara A Householder, Edward O List, John J Kopchick, Darlene E Berryman Ohio University, Athens, OH; Heritage College of Osteopathic Medicine, Ohio University, Athens, OH; Ohio University, Athens, OH Fibrosis refers to the formation of excess connective or scar tissue in an organ and is usually attributed to changes in the amount and composition of extracellular matrix (ECM) proteins. Growth hormone (GH), a protein secreted from the anterior pituitary, has repeatedly been shown to be positively correlated to ECM deposition in many tissues. The primary ECM protein in adipose tissue is collagen. Increases in collagen have been shown to be directly correlated with a decrease in adipocyte size. Generally, a reduction in adipocyte size would be considered a positive change. However, it has been suggested that it may have negative implications in carbohydrate and lipid metabolism as well as inflammation. To date, no study has assessed the role of GH in ECM deposition in adipose tissue. Bovine growth hormone transgenic mice (bGH) are giant and lean, yet have shortened life spans. In this study, the ECM deposition in adipose tissue was assessed in tissue collected from bGH versus littermate control mice at 10, 26, and 62 weeks of age, in both males and females. Adipose tissue samples from subcutaneous, perigonadal, retroperitoneal, and mesenteric depots were fixed and embedded in paraffin blocks and stained with pico-sirius red. When assessing overall collagen content, the bGH mice show greater ECM deposition when compared to the WT controls at most ages and in both sexes. Furthermore, adipocyte size was reduced in the bGH mice, with the subcutaneous depot showing the most dramatic reduction in size. When comparing different depots, collagen staining appeared most prevalent in the subcutaneous depot as compared to intraabdominal depots in both male and female mice. These results demonstrate a novel role of GH in adipose tissue metabolism and suggest that ECM deposition likely contributes to the depot specific differences in adipose tissue previously reported in mice with modified GH action.[br][br]Sources of Research Support: This work was supported in part by the State of Ohio[apos]s Eminent Scholar Program that includes a gift from Milton and Lawrence Goll, by the AMVETS, by the Diabetes Research Initiative at Ohio University, and by NIH grants DK083729, AG031736.[br][br]Nothing to Disclose: KMT, LAH, EOL, JJK, DEB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2043 52 210 SAT-97 PO27-02 Saturday 149 2012


150 ENDO12L_SAT-98 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Depot Specific White Adipose Tissue Immune Cell Populations in Male C57Bl/6J Mice Stephanie Harshman, Fabian Benencia, Riia Junnila, Edward O List, John J Kopchick, Darlene E Berryman Ohio University, Athens, OH; Heritage College of Osteopathic Medicine, Ohio University, Athens, OH; Ohio University, Athens, OH White adipose tissue (WAT) is a multifaceted organ composed of many cell types within the stromal-vascular fraction (SVF). An assortment of leukocytes is present in SVF including adipose tissue macrophages (ATMs), dendritic cells, and lymphocytes. These immune cells have been shown to have a role in fat tissue inflammation, glucose homeostasis, and insulin resistance through cytokine secretion and cell recruitment. Cell populations change in obese states and appear to alter the endocrine products secreted from adipose tissue and contribute to the inflammation commonly associated with obesity. Many of these general classes of immune cells can be further subdivided based on physiological function. Thus, it is important to not only evaluate the general classes of immune cells present in the tissue under a given condition but also to determine the subclasses in order to better appreciate their role in adipose tissue physiology. Adding to the complexity of adipose tissue are depot specific differences in cellular complexity and endocrine output. To date, little is known regarding the depot differences in immune cell populations or the influence of aging. Therefore, the purpose of this study was to characterize the depot specific differences of immune cell populations in relation to age. Adipose tissue from four distinct depots (inguinal, retroperitoneal, epididymal, and mesenteric) was collected from male C57Bl/6J mice at 5 months of age (young adult) and 21 months of age (aged adult). Dissected adipose tissue was treated immediately with collagenase and the SVF collected. The SVF samples were incubated with a series of fluorphore antibodies and analyzed by FACSAria II flow cytometer with FlowJo software to observe immune cell characteristics. The results demonstrate a unique inflammatory profile in the 21-month subcutaneous and mesenteric depots with significant macrophage and dendritic cell populations compared to retroperitoneal and epididymal depots. In addition, CD45+ cells were identified in subcutaneous, epididymal, and mesenteric depots indicating the presence of B-lymphocytes. The changes in immune cell composition at two age points provided a better understanding of the diverse functionality of immune cell populations, their depot-specific expression, and their potential role in adipose tissue inflammation.[br][br]Sources of Research Support: This work was supported in part by the State of Ohio[apos]s Eminent Scholar Program that includes a gift from Milton and Lawrence Goll, by the AMVETS, by the Diabetes Research Initiative at Ohio University, by a Student Enhancement Award from Ohio University, by an Endocrine Society Summer Fellowship, and by NIH grants DK083729, AG031736.[br][br]Nothing to Disclose: SH, FB, RJ, EOL, JJK, DEB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2099 52 211 SAT-98 PO27-02 Saturday 150 2012


151 ENDO12L_SAT-99 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Effects of Myostatin and Its Propeptide on Transdifferentiation of C2C12 Myoblast to Adipocytes Heng Wang, Ann Yang, Jinzeng Yang University of Hawaii at Manoa, Honolulu, HI; Huazhong Agricultural University, Wuhan, China Myostatin, a key member of TGF-[beta] superfamily in skeletal muscle, inhibits myoblast proliferation and differentiation. There are conflicting reports about the role of myosatin in adipoctyes. Myostatin inhibits preadipocyte differentiation, but promotes adipogeneisis in mesenchymal cells. We previously demonstrated inhibitory effects of myostatin propeptide on myostatin function in both transgenic mice (1-5) and in vivo recombinant protein administration studies (6). Transgenic over-expression of myostatin propeptide results in 76-152% increase in main individual muscles at 12 months of age. Administration of the propeptide to neonatal mice at the age of 11 and 18 days increase 13-25% muscle mass at the age of 57 days. The formations of skeletal myocytes, adipocytes and chondrocytes initially derive from the same multipotent mesodermal precursors during embryonic development. The plasticity of myoblasts to adipogenic and osteogenic lineage determinations is well established in vitro. To investigate the effects of paracrine effects of myostatin on adiposity in skeletal muscle, we initiate studying the effects of myostatin on transdifferentiation of C2C12 myoblast to adipocytes. Myostatin mRNA is expressed in a very low level in proliferating C2C12 cells. After successful transdifferentiation of C2C12 myoblasts into adipocyte-like cells by rosiglitazone and PPAR-[gamma] cDNA tranfection, we demonstrated that expressions of myostatin mRNA were induced by the adipogenic medium during myoblast adipogenesis. The relative level of myostatin mRNA by qPCR showed 20-fold increase compared with that in proliferating cells. The treatment with myostatin (1[mu]g/ml) throughout the transdifferentiation period enhanced adipogenesis as evidenced by the lipid accumulation in cell staining and increased expressions of adipogenic markers of PPAR-[gamma] and C/EBP[alpha]. However, little or no effect of myostatin propeptide on the transdifferentiation was observed. The supplementation of the propeptide at a higher level (5[mu]g/ml) showed inhibitory effects on adipogenesis. These results provide initial cell culture evidences that myostatin enhance adipogenesis during myoblast transdifferentiation, which suggest a different role of myostatin in adipogenesis in aged animals and humans when skeletal muscle is naturally regress to atrophy and accumulation of adipose tissue is prevailing.[br][br]1. Yang J, Ratovitski T, Brady JP, Solomon MB, Wells KD, Wall RJ. 2001. Mol Reprod Dev. 60:351-61. 2. Zhao B, Li EJ, Wall RJ, Yang J. 2009. BMC Genomics 10:305. 3. Yang and Zhao. 2006. Mol. Reprod. Dev. 73:462-9. 4. Zhao B, Wall RJ, Yang J. 2005. Biochem Biophys Res Commun. 337:248-55. 5. Suzuki ST, Zhao B, Yang J. 2008. Biochem Biophys Res Commun. 369:767-73. 6. Li Z, Zhao B, Kim YS, Hu CY, Yang J. 2010. Mol. Reprod. Dev. 77:76-82.[br][br]Sources of Research Support: USDA Hatch/Multistate-National Animal Genome Research Program HAW00244-R; Wuhan [quot]3551 Talent Scheme[quot].[br][br]Nothing to Disclose: HW, AY, JY 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 516 52 212 SAT-99 PO27-02 Saturday 151 2012


152 ENDO12L_SAT-100 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Anti-Inflammatory Effects of Full-Length and Globular Protein Fragments of the New Adipokine CTRP-3 Andreas Schmid, Andrea Kopp, Christa Buechler, Andreas Schaeffler University Hospital of Regensburg, Regensburg, Germany [bold]Introduction[/bold][br]C1q/TNF-related protein-3 (CTRP-3) is a newly discovered adipokine with anti-inflammatory impact on monocytes by down-regulation of pro-inflammatory cytokines, e.g. interleukine 6 (IL-6) and tumor necrosis factor alpha (TNF-alpha), in both adipocytes and monocytes. So far it is unclear which kind of interactions and molecular mechanisms are underlying the observed effects. Our aim is to characterize the differential effects of the globular and the collagenous domain of CTRP-3 and to compare these with the effects exerted by the full length protein.[br][bold]Methods[/bold][br]DNA of recombinant full-length CTRP-3, its globular domain, and its collagen domain were transfected into and expressed in H5 insect cells. The secreted proteins were extracted from the cell culture supernatant and purified by affinity chromatography.[br]Costimulation experiments were performed in 3T3-L1 and monocyte-like THP-1 cells with pro-inflammatory stimuli lipopolysaccharide (LPS) and free fatty acids (FFA) plus CTRP-3 or its globular domain, respectively. Gene expression and secretion of pro-inflammatory cytokines were measured by real time RT-PCR and ELISA techniques. Protein levels of intracellular components of pro-inflammatory signaling were analysed by Western blot.[br][bold]Results[/bold][br]Both full-length CTRP-3 and its globular domain were shown to decrease the amount of secreted IL-6, MCP-1 and resistin after costimulation with LPS and FFA in differentiated 3T3-L1 cells. RT-PCR results confirmed down-regulation of these pro-inflammatory factors on mRNA level. Decrease of secreted IL-6 in THP-1 cells upon costimulation with LPS and CTRP-3 globular domain could be shown as well.[br][bold]Conclusion and outlook[/bold][br]The results suggest that the observed anti-inflammatory impact of CTRP-3 is mediated by its globular domain, whereas the collagen like domain is still to be tested on these effects. As a next step we plan to investigate the anti-inflammatory effects of CTRP-3 in an animal model of LPS-induced SIRS.[br][br]Nothing to Disclose: AS, AK, CB, AS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 20 52 213 SAT-100 PO27-02 Saturday 152 2012


153 ENDO12L_SAT-101 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Mechanisms Underlying Adipocyte Dysfunction Caused by HIV-1 Viral Protein R (VPR) Dinakar Iyer, Neeti Agarwal, Toni Oplt, Ashok Balasubramanyam Baylor College of Medicine, Houston, TX The HIV-1 accessory protein viral protein R (Vpr) disrupts adipogenesis and adipocyte function in vitro. This is due in part due to co-repression of PPAR-gamma (1). Vpr also has other effects in many cell types, e.g., induction of cell cycle arrest at the G2/M phase (by sequestering Cdc25c and keeping the cyclin D/Cdk1 complex hyperphosphorylated); and promotion of apoptosis (by activating JNK, decreasing the ratio of Bcl2 to Bax, and binding to the mitochondrial ANT). We used Vpr mutants to investigate additional mechanisms of adipocyte dysfunction due to Vpr.[br]We used a tetracycline inducible lentivirus system in 3T3-L1 adipogenic cell lines to measure the effects of Vpr and the Vpr R80A mutant (which abolishes Vpr[apos]s ability to cause both G2/M cell cycle arrest and mitochondrial dysfunction, without affecting PPAR-gamma corepression). Synchronized 3T3-L1 preadipocytes were infected with known titers of control vector, Vpr or Vpr R80A constructs, induced for protein expression by doxycyline and allowed to undergo differentiation. Cells were harvested at different intervals (1 day to 10 days after induction), and expression levels of genes involved in adipogenesis were quantified by SyberGreen qPCR assay. Compared to vector-infected cells, Vpr expression induced a profound increase in mRNA expression of cyclins B and D at all time points, indicating altered cell cycle regulation, whereas expression of these cyclins in VprR80A expressing cells was not different from vector-infected controls. Vpr expression caused a global decrease (p[lt]0.01) in expression of genes critical for adipogenesis (PPAR[gamma], C/EBP[alpha], Foxo1) at all time points compared to uninfected or vector-infected cells, with a marked diminution of cells positive for Oil Red O after 10 days. By contrast, Vpr R80A expression permitted preadipocytes to progress through the phases of differentiation with temporally and quantitatively similar adipogenic gene expression as vector-infected cells, with a significantly greater fraction of Oil Red O-positive cells compared to Vpr treatment after 10 days.[br]Thus the Vpr R80A mutant substantially reduces Vpr[apos]s ability to disrupt adipocyte differentiation, indicating that cell cycle arrest, mitochondrial dysfunction, or both, play a prominent role in Vpr[apos]s antiadipogenic effect. Cellular measurements of mitochondrial respiration, glycolysis and fat oxidation are under way to dissect the mechanism.[br][br](1) Shrivastav S et al., 2007 22:234-247.[br][br]Nothing to Disclose: DI, NA, TO, AB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2031 52 214 SAT-101 PO27-02 Saturday 153 2012


154 ENDO12L_SAT-102 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Role of NAD[sup]+[/sup] Signaling through Poly(ADP-Ribose) Polymerase 1 in Adipogenesis Keun Woo Ryu, Xin Luo, William Lee Kraus University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX Adipose tissue plays a central role in the development of obesity and insulin resistance, which may involve exceeding the capacity of adipose tissue to expand and subsequent failure to recruit new adipocytes. Adipogenesis is tightly regulated by the sequential regulation of a set of key adipogenic transcription factors. Although the major transcription cascades and protein factors that regulate this process have been identified, our understanding of the precise molecular mechanisms of adipogenesis, particularly during the early stages of differentiation, is incomplete. However, recent studies have shown that nicotinamide adenine dinucleotide (NAD[sup]+[/sup])-dependent enzymes, including poly(ADP-ribose) polymerases (PARPs; e.g., PARP-1) play important roles in adipogenesis.[br]To elucidate the transcriptional regulatory functions of NAD[sup]+[/sup] signaling in adipogenesis, we are using a well-established model of adipogenesis: the murine 3T3L1 preadipocyte cell line. Using a variety of cell-based and molecular assays, we have found that (1) the poly(ADP- ribosyl)ation activity of PARP-1 fluctuates during adipogenesis; (2) RNAi-mediated depletion of PARP-1 alters adipogenesis-related gene expression and modulates the 3T3L1 cell differentiation program; (3) depletion of the nuclear NAD[sup]+[/sup] pools by RNAi-mediated knockdown of nicotinamide mononucleotide adenyltransferase 1 (NMNAT-1), a nuclear NAD[sup]+[/sup] synthase, reduces PARP-1 enzymatic activity enhances the differentiation of 3T3L1 cells; and (4) PARP-1 binds to the promoter regions of adipogenic target genes in a pattern that partially overlaps with the peroxisome proliferator activated receptor gamma (PPAR[gamma]), a key transcriptional regulator of adipogenesis. We are now using a variety of biochemical, molecular, cellular, and genomic assays to elucidate the exact role and regulatory mechanisms of NAD[sup]+[/sup] signaling in adipogenesis.[br]Collectively, these studies will help to elucidate the adipogenic regulatory network, as well as shed light on the mechanisms by which the nuclear NAD[sup]+[/sup] signal can regulate transcription to affect cell proliferation and differentiation. These studies have the potential to reveal new aspects of the pathogenesis of obesity and the potential therapeutic benefits of inhibitors for the NAD[sup]+[/sup]-dependent enzymes.[br][br]Sources of Research Support: This work is supported by a predoctoral fellowship from the DOD/BCRP to X.L. and a grant from the NIH/NIDDK to W.L.K.[br][br]Nothing to Disclose: KWR, XL, WLK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2090 52 215 SAT-102 PO27-02 Saturday 154 2012


155 ENDO12L_SAT-103 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Pyroglutamylated RF-Amide Peptide (QRFP) Gene Is Regulated by Lipopolysaccharide and Interferons in a Cellular Model of Metabolic Endotoxemia between Macrophages and Adipocytes Christian Jossart, Riccarda Granata, Marc G Servant, Sylvie Marleau, Huy Ong Universite de Montr[eacute]al, Montr[eacute]al, Canada; University of Turin, Turin, Italy Background: The orexigenic QRFP has been shown to be involved in feeding regulation. QRFP increases adipogenesis and inhibits lipolysis in 3T3-L1 adipocytes in a paracrine/autocrine manner. The expression of QRFP was found to be induced during differentiation of 3T3-L1 cells. However, in diet-induced obesity (DIO), a mouse model in which plasma concentrations of endotoxin have been shown to be slightly elevated, QRFP expression was decreased compared to lean animals. This metabolic endotoxemia (ME) seems to play a crucial role in the development of obesity. Objectives: 1) To document the mechanisms of regulation of QRFP gene. 2) To assess the effect of ME on QRFP expression in adipocyte and macrophage cell lines. Methods: The expression of QRFP in 3T3-L1 adipocytes and RAW264.7 macrophages was monitored by quantitative PCR following treatments with low doses of lipopolysaccharide (LPS) (1 ng/mL) corresponding to plasma concentration found in ME. TNF-[alpha] (10 ng/mL), palmitate (500 [micro]M) and interferon-beta (IFN-[beta], 100 U/mL) and gamma (IFN-[gamma], 10 ng/mL) have also been tested. Moreover, LPS-treated macrophages conditioned medium has been used on adipocytes. Signaling pathways have been investigated with shRNA knockdowns and pharmacological inhibitors. Results: We report the expression of QRFP in mouse peritoneal macrophages and in RAW and J774 macrophage cell lines. TNF-[alpha] and palmitate did not have any effect on QRFP expression. Low doses of LPS downregulated QRFP by 50% in macrophages but not in adipocytes. A direct, but transient, effect of LPS on QRFP expression in macrophages could be reversed with IKK-2 inhibitor IV or the knockdown of TRIF but not the knockdown of MyD88. Interferons are induced by LPS in macrophages. IFN-[beta] could reduce QRFP expression in both macrophages and adipocytes in similar magnitude than LPS but in a more sustained manner. IFN-[gamma] downregulated QRFP, but only in macrophages. Macrophage-conditioned medium reduced QRFP expression in adipocytes and this effect was blocked with an IFN-[beta] neutralizing antibody. The effect of IFN-[beta] on QRFP expression was blocked by PI3K inhibitor (LY294002) and by p38 MAPK inhibitor (SB203580). Conclusions: LPS induces IFN-[beta] release from macrophages which reduces QRFP expression in both macrophages and adipocytes in an autocrine/paracrine dependent manner, suggesting QRFP as a potential biomarker in ME.[br][br]Sources of Research Support: This work was supported by Aeterna Zentaris.[br][br]Nothing to Disclose: CJ, RG, MGS, SM, HO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 366 52 216 SAT-103 PO27-02 Saturday 155 2012


156 ENDO12L_SAT-104 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) GRK2 Contribution to the Regulation of Energy Expenditure and Brown Fat Function Sonia Fernandez-Veledo, Rocio Vila-Bedmar, Lucia Garcia-Guerra, Iria Nieto-Vazquez, Federico Mayor, Jr, Cristina Murga IISPV Universitat Rovira i Virgili, Tarragona, Spain; CSIC-UAM, Madrid, Spain; CSIC-UAM, Madrid, Spain; Complutense University, Madrid, Spain Obesity is a major health problem and an important risk factor for the development of multiple disorders. Previous studies in our laboratory have revealed that downregulation of GRK2 decrease age-related adiposity (1), but the physiological and molecular mechanisms underlying this outcome remain unclear. We evaluate whether the lean phenotype results from a direct effect of GRK2 on energy homeostasis. The study of WAT in WT and GRK2+/- littermates showed a reduced expression of lipogenic enzymes and enhanced lypolitic rate in adult GRK2+/- mice. Moreover, hemizygous mice display higher energy expenditure and lower respiratory exchange ratio. Analysis of BAT from adult GRK2+/- mice showed a less deteriorated morphology associated to age compared to WT, which is correlated with a higher basal core temperature. BAT from young GRK2+/- mice showed an increase in gene expression of thermogenesis-related genes. Accordingly, hemyzigous mice display better thermogenic capacity and exhibited a more oxidative phenotype in both BAT and WAT than WT littermates. Overexpression of GRK2 in brown adipocytes corroborated the negative impact of this kinase in BAT function and differentiation. Collectively, our data point at GRK2 inhibition as a potential tool for the enhancement of brown fat activity, which may have important therapeutic implications for the treatment of obesity and associated metabolic disorders.[br][br](1) Garcia-Guerra L et al., Diabetes 2010; 2407-17.[br][br]Sources of Research Support: Ministerio de Educacion y Ciencia (SAF2008-00552 to F.M and BFU2008-04043 to S.F-V); Fundacion Ramon Areces; Comunidad de Madrid (S-SAL-0159-2006);Ministerio Sanidad y Consumo-Instituto Carlos III via The Cardiovascular Network (RECAVA, RD06-0014/0037 to F.M. and PS09/0128 to C.M) and Fondo de Investigacion Sanitaria (FIS) [ldquo]Miguel Servet[rdquo] Grant CP10/00438 to S.F-V.[br][br]Nothing to Disclose: SF-V, RV-B, LG-G, IN-V, FM, CM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1137 52 217 SAT-104 PO27-02 Saturday 156 2012


157 ENDO12L_SAT-105 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) The Expression in and Role of Oxytocin Receptor on Adipose Tissue Lipid Metabolism under Different Feeding Conditions in Mice Sanggun Roh, Kyoungha So, Yutaka Suzuki, Chen Chen, Kazuo Katoh Tohoku University, Sendai, Japan; University of Queensland, Brisbane, Australia Oxytocin, synthesized in hypothalamic neurons, transported to the posterior pituitary gland and released into peripheral circulation, cannot re-enter the brain because of the blood-brain barrier. Through specific and high affinity oxytocin receptor (OXTR), the behavioral effects of oxytocin are thought to reflect only release from centrally projecting oxytocin neurons, separated from hypothalamic ones that project to the posterior pituitary gland. Peripheral actions of oxytocin are mediated by OXTR, which ubiquitously expressed in peripheral tissues including mammary gland. It has been reported that oxytocin intraperitoneal injection induced lipolysis and improved glucose tolerance. However, the role of changing OXTR expression in adipose tissue to have balanced catabolic effects under different metabolic conditions is not clear. The aim of this study was to investigate the expression of OXTR in adipose tissues and possible role in regulating lipid metabolism. To clarify whether OXTR was expressed in isolated adipocytes, its RNA level was analyzed in adipocytes and stromal-vascular cells isolated from mouse adipose tissues. OXTR was highly expressed in adipocytes, with a much lower level of expression in stromal-vascular cells. OXTR mRNA expression was markedly up-regulated in adipose tissues of mice fed a high-fat diet compared with those fed a normal-fat diet. However, OXTR mRNA expression was down-regulated in pituitary gland of mice fed by a high-fat diet. In addition, OXTR mRNA was significantly increased in adipose tissues of db/db mice compared with wild type. To investigate differential gene expression of OXTR in adipose tissue in response to changes in nutritional status, mice was deprived of feeding for 16 hr and then allowed to refeed for 4hr. OXTR gene expression was markedly reduced in adipose tissue of mice deprived of feeding compared with ad libitum fed lean mice, but was increased after refeeding. To investigate whether oxytocin modified the expression of OXTR genes, 3T3-L1 adipocytes were treated with oxytocin (100 pg/ml) for 24 hr. Oxytocin treatment significantly up-regulated OXTR mRNA. These results suggest that up-regulation of OXTR during fat accumulation and adipocyte development may intensify the catabolic effects by oxytocin in adipose tissue to cause lipolysis.[br][br]Sources of Research Support: Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan.[br][br]Nothing to Disclose: SR, KS, YS, CC, KK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1573 52 218 SAT-105 PO27-02 Saturday 157 2012


158 ENDO12L_SAT-106 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Paradoxical Changes in Glucose Tolerance with Aging in Mice Over-Expressing Human IGFBP3: Possible Role of Brown Fat Hoa Khanh Nguyen, Xing Hai Yao, Suresh Mishra, Gregoire Nyomba University of Manitoba, Winnipeg, Canada; University of Manitoba, Winnipeg, Canada [bold]Introduction:[/bold] Three month-old transgenic (Tg) mice overexpressing human IGFBP3 (BP3) and mutant IGFBP3 (mBP3) lacking IGF binding ability have different degrees of glucose intolerance. In this study we have investigated the role of brown fat in the glucose intolerance. [bold]Methods:[/bold] Glucose status of 3, 6 and 12 month-old BP3 and mBP3 mice was studied by glucose (GTT) and insulin (ITT) tolerance tests, and expression of brown fat marker mRNAs was determined by real time PCR in the interscapular fat tissue. [bold]Results:[/bold] At 3, 6 and 12 months, respectively, body weight was 36.5[plusmn]1.0, 42.4[plusmn]1.2, 41.1[plusmn]0.8g in BP3; 40.2[plusmn]0.8, 45.4[plusmn]1.5, 50.1[plusmn]1.3g in mBP3; 42.7[plusmn]1.0, 48.4[plusmn]1.4, 53.3[plusmn]1.9g in wild-type (WT) mice. Glucose AUC was 1726[plusmn]93[sup][amp][/sup], 1854[plusmn]91, 1548[plusmn]146 in BP3; 1743[plusmn]76[sup][amp][/sup], 2241[plusmn]137[sup]#[/sup], 2131[plusmn]117[sup]#[/sup] in mBP3; 1413[plusmn]32, 1442[plusmn]92, 1382[plusmn]57 in WT mice. K[sub]itt[/sub] was 3.66[plusmn]0.42, 4.16[plusmn]0.42[sup][amp][/sup], 4.03[plusmn]0.24 in BP3; 4.14[plusmn]0.70[sup][amp][/sup], 3.21[plusmn]0.5, 0.98[plusmn]0.6[sup][sect][/sup] in mBP3; 2.34[plusmn]0.42, 2.98[plusmn]0.36, 3.35[plusmn]0.29 in WT mice. Percent brown fat was 0.48[plusmn]0.04, 0.52[plusmn]0.04, 0.51[plusmn]0.04 in BP3; 0.55[plusmn]0.03, 0.80[plusmn]0.4[sup][amp]*[/sup], 0.95[plusmn]0.06[sup][sect]$[/sup] in mBP3; 0.55[plusmn]0.03, 0.58[plusmn]0.05, 0.67[plusmn]0.06 in WT mice. Fat accumulation was greater in the interscapular space of mBP3 vs. BP3. At 6 months, mRNA increased for uncoupling protein-1 (UCP1; 6.16[plusmn]1.59[sup][sect][/sup] vs. 1.25[plusmn]0.19), peroxisome proliferator-activated receptor-[gamma] (PPAR[gamma]; 3.90[plusmn]0.56[sup][amp][/sup] vs. 1.62[plusmn]0.40), PPAR-coactivator-1[alpha] (PGC1[alpha]; 3.45[plusmn]0.57[sup][sect][/sup] vs. 1.30[plusmn]0.16), PGC1[beta] (5.63[plusmn]1.29[sup][amp][/sup] vs. 1.45[plusmn]0.30), Twist (3.53[plusmn]0.80[sup][sect][/sup] vs. 0.99[plusmn]0.11) and PRDM16 (3.72[plusmn]0.52[sup][sect][/sup] vs. 1.18[plusmn]0.10) in BP3 vs. WT mice; mBP3 had increases for PGC1[alpha] (2.75 [plusmn]0.55[sup][amp][/sup]), PGC1[beta] (7.72[plusmn]2.01[sup][sect][/sup]), Twist (3.95[plusmn]0.45[sup][sect][/sup]) and PRDM16 (4.15[plusmn]0.89[sup][sect][/sup]), but not PPAR[gamma] (3.08[plusmn]0.62) or UCP1 (3.25[plusmn]1.09). At 12 months, vs. WT mice, UCP1 (2.63[plusmn]0.42[sup][amp][/sup] vs. 1.11[plusmn]0.09) mRNAs were higher in BP3, while PGC1[alpha] (0.78[plusmn]0.18[sup][amp][/sup] vs. 1.35[plusmn]0.36), PPAR[gamma] (0.78[plusmn]0.11[sup][micro][/sup] vs. 1.17[plusmn]0.14) and PRDM (0.57[plusmn]0.07[sup][amp]*[/sup] vs. 1.01[plusmn]0.07) mRNAs were lower in mBP3. [bold]Conclusion:[/bold] Glucose intolerance occurred in both Tg mice, but was worse in mBP3, and improved with age in BP3 but not in mBP3, which developed insulin resistance with age. Thus, mutant IGFBP3, which does not bind IGFs, led to insulin resistance with reduced expression of brown fat specific genes, whereas these genes were higher in the presence of native IGFBP3. This suggests that brown fat explains differences in insulin sensitivity between the two Tg mice via IGF dependent and independent mechanisms.[br][sup][amp][/sup]p[lt]0.05,[sup][sect][/sup]p[lt]0.01, [sup]#[/sup]p[lt]0.001 vs. WT mice;[sup][micro][/sup]p[lt]0.05, [sup]*[/sup]p[lt]0.01,[sup]$[/sup]p[lt]0.001 vs. BP3 mice.[br][br]Sources of Research Support: The study was supported by a grant from the Canadian Institutes of Health Research (MOP-64306).[br][br]Nothing to Disclose: HKN, XY, SM, GN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 73 52 219 SAT-106 PO27-02 Saturday 158 2012


159 ENDO12L_SAT-107 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Fas (CD95) Induces Lipolysis in 3T3-L1 Adipocytes Via Activation of the ERK1/2 Pathway Reto A Rapold, Stephan Wueest, Adrian Knoepfel, Eugen J Schoenle, Daniel Konrad University Children[apos]s Hospital, Zurich, Switzerland; University Children[apos]s Hospital, Zurich, Switzerland; University of Zurich, Zurich, Switzerland [bold]Background and Aims[/bold]: Fas (CD95) belongs to the superfamily of the tumor necrosis factor (TNF) receptors. Besides its key role in apoptosis, Fas contributes to non-apoptotic pathways such as cell proliferation and inflammation. We previously found that Fas-deficient and adipocyte-specific Fas knockout mice are partly protected from high fat diet-induced insulin resistance. The aim of the present study was to elucidate the underlying molecular mechanisms of this protective effect; in particular the impact of Fas activation on lipolysis in adipocytes.[br][bold]Materials and Methods:[/bold] Fully differentiated 3T3-L1 adipocytes were treated with 2 ng/ml membrane-bound Fas ligand (FasL) in the presence or absence of the MEK inhibitor U0126, the Ca2+ chelator BAPTA/AM, the Ca2+/calmodulin dependent protein kinase II (CaM kinase II) inhibitor KN62 or after siRNA mediated p44/42 MAP kinases (ERK1/2) depletion. Protein was collected for western blot and Luminex analysis. Lipolysis was assessed by measuring free glycerol and free fatty acid release.[br][bold]Results:[/bold] Treatment of adipocytes with FasL induced lipolysis in a time dependent manner with an approximately 2-fold increase after 12 hours. In parallel, Fas activation increased phosphorylation of ERK1/2. FasL-induced lipolysis was blunted in the presence of the ERK-inhibitor U0126 or in ERK1/2 depleted adipocytes. In addition, blocking of the Ca2+/calmodulin kinase pathway (either by a Ca2+ chelator or by a CaM kinase II inhibitor) significantly blunted FasL-induced ERK1/2 phosphorylation and glycerol release.[br][bold]Conclusions[/bold]: Fas activation in mature 3T3-L1 adipocytes induces lipolysis in a CaM kinase II/ERK1/2-dependent manner and, thus, may impact on adipocyte and whole-body metabolism.[br][br]Sources of Research Support: Swiss National Science Foundation.[br][br]Nothing to Disclose: RAR, SW, AK, EJS, DK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 470 52 220 SAT-107 PO27-02 Saturday 159 2012


160 ENDO12L_SAT-108 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Effects of Growth Differentiation Factor 3 (GDF3) on Differentiation and Mature Adipocyte Function Maria Namwanje, Chester Brown Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX Growth differentiation factor 3 (GDF3) is a member of the TGF-[beta] family expressed in adipose tissue and its expression is up regulated under high fat diet (HFD) conditions. Gdf3 knock out mice are protected against HFD-induced obesity compared to wild type mice. In contrast, adenoviral transfer of GDF3 in mice results in enhanced sensitivity to HFD including increased adipocyte size and adipose tissue mass relative to wild type mice under the same diet conditions. These results suggest a possible role of GDF3 in regulation of differentiation and/or function of adipocytes. We hypothesize that under HFD conditions, loss of Gdf3 function results in increased lipolysis via effects on [beta]-adrenergic receptor signaling or other mechanisms, or defects in lipogenesis resulting in protection from obesity. We differentiated 3T3-L1 cells into mature adipocytes and treated the cells with GDF3 recombinant protein. We used quantitative RT-PCR to assess the expression of mature adipocyte genes and glycerol and non-esterified fatty acid assays to measure lipolytic activity. GDF3 had no significant effect on the amount of free glycerol and non-esterified fatty acids released at basal and stimulated levels in 3T3-L1cells. Also GDF3 has no effect on expression of lipogenic and lipolytic genes.[br]We then examined the cell autonomous effects of GDF3 on differentiated mouse embryonic fibroblasts (MEFs) from wild type and Gdf3-/- mice. On the whole the Gdf3-/- MEFs had a higher expression of Ppar[gamma] and Prmd16 compared to the wild type cells at day 2 and day 4 after induction of differentiation. The expression of Cebp-[alpha], an upstream regulator of Ppar[gamma] during adipogenesis, was not affected during the differentiation of wild type and Gdf3-/- MEFs. We are currently examining the effects of GDF3 on cell proliferation and apoptosis in MEFs as well as primary preadipocytes.[br][br]Nothing to Disclose: MN, CB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1001 52 221 SAT-108 PO27-02 Saturday 160 2012


161 ENDO12L_SAT-109 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Different Responses to Growth Differentiation Factor 3 in Adipogenic Cell Lines Juan Bournat, Chester Brown Baylor College of Medicine, Houston, TX Growth differentiation factor 3 (GDF3) is a member of TGF-beta superfamily whose expression in white adipose tissues is induced by high fat diet (HFD). Body weight and adipose tissue mass normally increase under HFD conditions, and these processes are augmented when adenovirus-mediated gene transfer overexpresses GDF3 systemically. In contrast, Gdf3 knockout mice have less adipose tissue mass than wild type mice under HFD conditions and exhibit higher basal metabolic rates. These results suggest that GDF3 may act as an adipogenic factor under conditions of caloric excess, and that GDF3 deficiency results in the dysregulation of energy expenditure. Interestingly, the brown adipose markers, CPT1b, PGC1-alpha, and UCP1 are overexpressed selectively in the white adipose tissues of GDF3 knockout mice on HFD, suggesting that GDF3 may play a role in modulating brown versus white adipocyte fate decisions. However, the signaling pathway(s) that mediate these effects are unknown. Recent reports indicate that GDF3 binds BMPs and inhibits their signaling during development. BMPs play important roles in adipogenesis, and BMP7 can induce brown adipocyte differentiation. Moreover, myostatin (GDF8) can inhibit adipogenesis by selectively inhibiting BMP7 signaling. Therefore, we are studying the molecular mechanisms underlying the effects of GDF3 alone and on BMP-dependent adipogenesis using pre-adipocyte cell lines, reporter assays, and quantitative PCR (qPCR). Our results show that GDF3 increases the proliferation of 3T3-L1 a committed white pre-adipocyte cell line, but does not affect the proliferation of C3H10T1/2 a multipotent mesenchymal stem cell (MSC) line. We also find that GDF3 selectively blocks both BMP4 and BMP7 signaling in 3T3-L1 cells, however, GDF3 does not have an effect on BMP4 or BMP7 signaling in C3H10T1/2 cells. In addition, GDF3 alone or in combination with BMPs increases the expression and activity of PPAR-gamma in these preadipocyte cell lines. These observations could be due to intrinsic differences between the cells that affect the expression of signaling components (either present or absent) that contribute to the inhibition of BMP signaling, such as receptors, ligands, co-receptors/ligands, extracellular inhibitors, or downstream mediators of canonical or non-canonical signaling pathways, and have provided insight into the stages and mechanisms by which GDF3 can influence adipogenesis.[br][br]Sources of Research Support: This work was supported by National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) RO1 Grant DK073572 to Dr. Chester Brown. Dr Juan Bournat was supported by a research supplement to promote diversity in health related research from the NIDDK.[br][br]Nothing to Disclose: JB, CB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 992 52 222 SAT-109 PO27-02 Saturday 161 2012


162 ENDO12L_SAT-110 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Trichostatin A (TSA) Modulates TZD Suppression of TNF[alpha]-Induced Lipolysis in Adipocytes Juu-Chin Lu, Yu-Tzu Chang, Chun-Ken Lin, Zhong-Sheng Wu Chang Gung University, Tao-Yuan, Taiwan; Chang Gung University, Tao-Yuan, Taiwan; Chang Gung University, Tao-Yuan, Taiwan Tumor necrosis factor [alpha] (TNF[alpha]) has several effects on adipocytes that may contribute to the development of insulin resistance. It increases local inflammation in adipose tissue and induces basal lipolysis of adipocytes, resulting in elevated serum levels of proinflammatory cytokines, chemokines, and free fatty acids, which have been suggested to mediate pathogenesis of obesity-linked chronic diseases such as insulin resistance and type 2 diabetes mellitus. The peroxisome proliferator-activated receptor [gamma] (PPAR[gamma]) is a nuclear receptor and the key protein controlling adipocyte function. It is also the cellular target of insulin-sensitizing drug thiazolidinediones (TZDs), the synthetic agonists of PPAR[gamma] used in treating diabetic patients. TZD activation of PPAR[gamma] has been shown to antagonize several TNF[alpha] actions in adipocytes, including TNF[alpha]-induced lipolysis, which may account for parts of the insulin-sensitizing effects of TZDs. As a transcriptional factor, the action of PPAR[gamma] is closely modulated by coregulators which include coactivators and corepressors. Previous studies have revealed that in macrophages, insulin-sensitizing effect of PPAR[gamma] may involve suppression of proinflammatory gene expression by recruitment of the corepressor complex that contains corepressors and histone deacetylases (HDACs). However, the role of corepressor complex in adipocytes remains largely unknown. We use a pan-HDAC inhibitor trichostatin A (TSA) to determine the role of HDACs in TZD suppression of TNF[alpha]-induced lipolysis in adipocytes. Treatment of 3T3-L1 adipocytes with TSA increased basal lipolysis in adipocytes. Moreover, inhibition of HDAC activity by TSA also reduced the suppression effect of TZD on TNF[alpha]-induced lipolysis. To determine how TSA modulates TZD suppression on TNF[alpha]-induced lipolysis, downstream signaling molecules involved in TNF[alpha]-induced lipolysis were examined. While TNF[alpha] induced ERK1/2 MAPK phosphorylation, known to be involved in TNF[alpha]-induced lipolysis, treatment of TZDs suppressed TNF[alpha]-induced phosphorylation of ERK1/2. Interestingly, treatment with TSA not only increased basal phosphorylation of ERK1/2, but also reduced TZD suppression on TNF[alpha]-induced phosphorylation of ERK1/2. Further experiments will be required to elucidate possible mechanisms, which may improve therapeutic approaches in the future.[br][br]Nothing to Disclose: J-CL, Y-TC, C-KL, Z-SW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 749 52 223 SAT-110 PO27-02 Saturday 162 2012


163 ENDO12L_SAT-111 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Alendronate Inhibits Adipocyte Differentiation in 3T3-L1 Cells through a Vitamin D Receptor[ndash]Mediated Effect Massimiliano Caprio, Matilde Calanchini, Caterina Mammi, Antonella Antelmi, Francesca Cinti, Giuseppe Rosano, Andrea Fabbri IRCCS San Raffaele Pisana, Rome, Italy; University [ldquo]Tor Vergata, Rome, Italy BACKGROUND: Adipocyte and osteoblast derive from the same mesenchimal progenitor. Age-related decrease in bone mass is accompanied by an increase in marrow adipose tissue. Vitamin D3 (VD) inhibits adipogenesis in 3T3-L1 preadipocytes. Recently it has been demonstrated that Alendronate (ALN) inhibits adipogenesis while promoting osteoblast differentiation of mesenchimal stem cells.[br]AIM OF THE STUDY: To evaluate the in vitro role of ALN on 3T3-L1 adipose differentiation and the potential sinergic role of VD co-treatment.[br]PROCEDURES: Murine 3T3-L1 and 3T3-F442A preadipocytes were routinely differentiated for 7 days adding ALN and VD 10-9,10-8 and 10-7M, then stained with Red Oil. We analyzed through RT-PCR the effect of such treatments upon mRNA expression of main molecular markers of differentiation, PPAR[gamma] and C/EBP[alpha], and VD Receptor (VDR).[br]RESULTS: ALN displayed a marked anti-adipogenic effect on 3T3-L1 cells. VD showed a clear dose-dependent anti-adipogenic effect. Interestingly co-incubation of ALN 10-8M and VD 10-9M did not show synergic effect in inhibition of adipogenesis. PPAR[gamma] mRNA expression was significantly reduced by ALN and VD. mRNA expression of C/EBP[alpha] was reduced only by the highest doses of VD. Interestingly a concomitant increase in VDR mRNA expression was observed in the presence of ALN and VD, suggesting that VDR may represent the molecular target of the anti-adipogenic effect of ALN.[br]To confirm this hypothesis, we explored the effects of ALN and VD on 3T3-F44 cells, which are in a more advanced differentiation stage in adipogenesis than 3T3-L1 cells; interestingly, we found that expression of VDR mRNA was much lower in 3T3-F44 than in 3T3-L1 cells. Moreover, adipose differentiation in this cell model was not affected by ALN nor by VD, differently from what observed in 3T3-L1 cells.[br]CONCLUSIONS: These data represent an indirect evidence of the role of VDR in mediating the anti-adipogenic effect of ALN. Further studies are required to clarify this mechanism.[br][br]Nothing to Disclose: MC, MC, CM, AA, FC, GR, AF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 802 52 224 SAT-111 PO27-02 Saturday 163 2012


164 ENDO12L_SAT-112 POSTER SESSION: The Adipocyte [amp] Biology of Adipose Tissue (1:30 PM-3:30 PM) Cheonghunhwadam-Tang Inhibits Differentiation to Adipocytes in 3T3-L1 Mouse Fibroblasts Hoyoung Lee, Ji-Hye Lim, Myeong Soo Lee, Min-Ho Cha Korea Institute of Oriental Medicine, Deajeon, Republic of Korea Cheonghunhwadam-tang(CHT), a herbal formula, is have been used in oriental medicine for many centuries as a therapeutic agent of fire and phlegm. It has been composed with 16 herbs including Pinellia ternate (Thunb.) Breit, Citrus unshiu Markovich, Poria cocos Wolf and Atractylodes macrocephala Koidzumi etc. In this study, we examined the efficacy of CHT on adipogenesis. 3T3-L1 cells were differentiated for adipocytes given 1 mg/mL insulin, 0.5 mM isobutylmethylxanthine (IBMX) and 0.25 [mu]M Dexamethasone (DEX) for 7days. After differentiation, cells were stained with Oil-Red-O to detect oil droplets in adipocytes and measured for triglyceride (TG) and glycerol-3-phosphate dehydrogenase (GPDH) activity. We confirmed that PPAR-[gamma], C/EBP-[alpha], Fas and aP2 mRNA expression by RT-PCR and analyzed the gene by microarray in treatment of 3T3-L1 with CHT.[br]These finding showed that CHT suppressed GPDH activity and TG accumulation in a dose-dependent manner significantly. Also, the treatment of CHT reduced the levels of PPAR-[gamma], C/EBP-[alpha], Fas and aP2 mRNA. In microarray analysis, the most down/up expressed pathways were cell cycle and lipid regulation signaling. These results suggest that CHT may prevent obesity and abdominal obesity.[br][br]1) Marie Thompson G etal., Anal Biochem 2004; 21-28. 2) Gregory R etal., Gene 2004; 167-185. 3) Tersuro O etal., Food Chem 2010; 239-244.[br][br]Sources of Research Support: This research was supported by a grant from the Korea Institute of Oriental Medicine (K12130).[br][br]Nothing to Disclose: HL, J-HL, MSL, M-HC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 635 52 225 SAT-112 PO27-02 Saturday 164 2012


165 ENDO12L_SAT-113 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Body Mass Index and Age Determine the Acrophase of Prolactin in the Human Ferdinand Roelfsema, Hanno Pijl Leiden University Medical Center, Leiden, Netherlands Prolactin has many effects in animals and man. For example, it regulates fat mass in fish, birds and mammals. In particular, the timing of the serum PRL acrophase in relation to the light-dark cycle, or to serum cortisol in constant light conditions, determines whether the fat mass increases or decreases, as part of the adaptation to seasons (1). The role of PRL in this respect has been less well studied in man. Hyperprolactinemic patients have increased fat which decreases after bromocriptine. Other studies, in small groups, have demonstrated (slightly) increased PRL levels in obese subjects, which may be attributed to defective D2R signaling (2), but no detailed studies on PRL acrophase in large groups are available, nor the effect of age on the phase setting. We hypothesized that the timing of the PRL acrophase may be correlated with fat mass (or BMI as proxy) in the human, as observed in animals. We therefore retrospectively analyzed our 24h blood sampling studies in healthy volunteers, in whom both PRL and cortisol were measured. Seventy-four subjects were available, mean age 43 yr (range 22-77 yr) and BMI 26.8 kg/m[sup]2[/sup] (range 18.7-38.4 kg/m[sup]2[/sup]).The PRL acrophase occurred at 03:22 h [plusmn] 13 min (mean [plusmn] SEM) and that for cortisol at 10:03 h [plusmn] 15 min. In the forward selection multivariable regression analysis BMI (P=0.01) and age (P=0.012) were independent negative predictors of the acrophase, but gender was not a significant factor. The cortisol acrophase correlated negatively with age (R=0.308, P[lt]0.009), but not with BMI or gender. The time difference between the cortisol and PRL acrophase was positively correlated with BMI (R=0.418, P[lt]0.001). In various species a wide gap between the cortisol acrophase and that of PRL leads to fat storage. Our findings are consistent with this observation, although we had to use BMI as proxy. If an advance shift of the PRL acrophase in relation to that of cortisol is also responsible for increased fat mass in man, manipulating the PRL phase may open an alternative route for treating obesity.[br][br](1)Meier AH and Cincotta AH, Diabetes Rev 4:464-87, 1996. (2)Kok P et al, JCEM 89:4445-9, 2004.[br][br]Nothing to Disclose: FR, HP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 34 53 226 SAT-113 PO28-02 Saturday 165 2012


166 ENDO12L_SAT-114 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Regulation of Adiponectin by Free Fatty Acids in Lean and Obese Subjects Kyung Wook Kim, Sy Nam, CW Ahn, KR Kim, JM Yu, Bjoern Richelsen Yongin Severance Hospital, Yongin-si Gyeonggi-do, Republic of Korea; Gangnam Severance Hospital, Seoul, Republic of Korea; Hallym University Kangnam Medical Center, Seoul, Republic of Korea; Aarhus University Hospital, Aarhus, Denmark [italic]Objective[/italic]: Among the adipocyte-derived cytokines, adiponectin possesses an enhancer of insulin sensitivity, anti-inflammatory and antiatherogenic properties. It is suggested that adiponectin plays an important role in lipid metabolism, however, whether free fatty acids (FFA) regulate adiponectin levels remains unclear. The present study was thus undertaken to evaluate the effects of FFAs on the regulation of plasma adiponectin and adipose tissue adiponectin mRNA expression in lean and obese subjects.[br][italic]Methods[/italic]: Seven obese men (BMI 31.3 [plusmn] 1.1 kg/m[sup]2[/sup]) and eight lean men (BMI 21.3 [plusmn] 0.4 kg/m[sup]2[/sup]) were studied. Subjects received an Intralipid 20% (Greencross, Korea) infusion at a rate of 1.5 mL/min over 180-minute period together with sodium heparin at a rate of 1600 IU/h (priming dose 200 IU). Blood sampling and abdominal subcutaneous adipose tissue biopsy through percutaneous mini-liposuction were taken before and after 180-minute Intralipid/heparin infusion.[br][italic]Results[/italic]: The FFA concentration was increased 6 fold after Intralipid/heparin infusion in both lean and obese groups. Adipose tissue adiponectin mRNA was significantly reduced by Intralipid/heparin infusion (38.0 [plusmn] 7.7 [italic]vs.[/italic] 21.1 [plusmn] 3.0.0, [italic]p[lt]0.05[/italic]), and the plasma level of adiponectin was reduced from 5.72 to 5.37 which, however, was a non-significant reduction ([italic]p=0.16[/italic]). If the two groups were separated, in the lean subjects, adipose tissue adiponectin mRNA was significantly lower after Intralipid/heparin infusion (48.3 [plusmn] 12.5 [italic]vs.[/italic] 23.2 [plusmn] 5.0, [italic]p[lt]0.05[/italic]), whereas there was no difference in plasma adiponectin before and after (5.89 [plusmn] 0.6 [italic]vs.[/italic] 5.84 [plusmn] 0.8, [italic]p=0.88[/italic]). In obese subjects, plasma adiponectin concentration was significantly lower after Intralipid/heparin infusion (5.5 [plusmn] 0.8 [italic]vs.[/italic] 4.8 [plusmn] 0.8, [italic]p[lt]0.05[/italic]), whereas there was no difference in adipose tissue adiponectin mRNA expression (26.0 [plusmn] 6.3 [italic]vs.[/italic] 18.7 [plusmn] 3.1, [italic]p=0.24[/italic]).[br][italic]Conclusions[/italic]: Albeit the other data on the effect of FFAs on adiponectin concentrations are not consistent, we could get that FFAs suppress the expression of adiponectin in lean and obese human. The issue of possible FFA-induced regulation of adiponectin warrants further investigations.[br][br]Nothing to Disclose: KWK, SN, CWA, KRK, JMY, BR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 59 53 227 SAT-114 PO28-02 Saturday 166 2012


167 ENDO12L_SAT-115 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Morbidly Obese Hypothyroid Patients Require an Increase in Weight-Based Thyroid Hormone Dose Following Bariatric Surgery Umasankari Sundaram, Valerie K Shostom, Jane L Meza, Corrigan L McBride, Melissa M Monzu-Sparks, Whitney S Goldner University of Nebraska Medical Center, Omaha, NE; University of Nebraska Medical Center, Omaha, NE; University of Nebraska Medical Center, Omaha, NE [bold]Background[/bold][br]The relationship between morbid obesity and thyroid hormones is complex and obesity itself can be accompanied by changes in thyroid function. In addition, hypothyroidism is common in morbidly obese patients. Previous studies have been inconsistent and have reported an increase, decrease, and no change in total thyroid hormone dose following bariatric surgery. We sought to evaluate the change in thyroid hormone dose following bariatric surgery in hypothyroid morbidly obese patients and the factors that influenced the change.[br][bold]Methods[/bold][br]We retrospectively analyzed 231 hypothyroid morbidly obese patients on levothyroxine replacement (LT4) who underwent Roux-en-Y-gastric bypass, lap banding or sleeve gastrectomy at The University of Nebraska Medical Center(UNMC) between January 2000 and December 2010. We analyzed for change in LT4 dose (both actual dose and weight based dose (mcg/kg)) and correlation between change in dose and confounding variables including change in weight, BMI, TSH, and vitamins B12 and 25OHD.[br][bold]Results[/bold][br]The prevalence of hypothyroidism in morbidly obese patients undergoing bariatric surgery at UNMC was 19% (231/1221). Complete pre and post-operative thyroid data was available on 74 patients. Overall, 19/74 (26%) had a decrease, 41/74 (55%) had no change, and 14/74 (19%) had an increase in actual LT4 dose. Pre and post-operative median TSH was not significantly different (pre: 2.64 mcIU/ml, post: 2.0 mcIU/ml) and median LT4 dose was also not different pre and post-operatively (125 mcg for both) between groups. However, the weight based LT4 dose following surgery was significantly increased post-operatively (pre: 0.95 mcg/kg) versus (post: 1.14 mcg/kg) (p=[lt]0.01). There was no significant difference between patient[apos]s minimum vitamin B12 and 25OHD levels and change in actual and weight based dose, however, there was a trend for the lowest levels of B12 and 25OHD being inversely associated with both actual and weight based LT4 dose.[br][bold]Conclusion[/bold][br]We observed an increase in weight based LT4 dose following bariatric surgery. However, actual LT4 dose changes were variable, and the majority had no change. This suggests malabsorption as a major contributor to post-operative changes in thyroid hormone dose rather than weight loss alone as the main contributor to thyroid hormone dose changes following bariatric surgery.[br][br]Nothing to Disclose: US, VKS, JLM, CLM, MMM-S, WSG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 234 53 228 SAT-115 PO28-02 Saturday 167 2012


168 ENDO12L_SAT-116 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Testosterone Therapy with Testosterone Undecanoate Long-Acting Intramuscular Injections in Hypogonadal Men Is Associated with Reduction in Obesity Aksam Yassin, Youssef El Douaihy, Ridwan Shabsigh, Aiman Yassin, Farid Saad Segeberger Kliniken, Norderstedt, Germany; Dresden International University, Dresden, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates; Maimonides Medical Center, Brooklyn, NY; Klinikum Braunschweig, Braunschweig, Germany; Bayer Pharma AG, Berlin, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates [bold]Introduction[/bold]: Obesity and the metabolic syndrome are frequently associated with late onset hypogonadism (LOH). We sought to study the effects of testosterone replacement therapy on weight, body mass index (BMI) and waist circumference (WC) in patients with LOH.[br][bold]Materials and Methods[/bold]: As of November 2004 130 patients with LOH (diagnosis criteria: total testosterone [le]3.5ng/dl and documented erectile dysfunction (ED) according to Sexual Health Inventory for Men score (SHIM; [le] 21) were included in a prospective cohort study to investigate the effect of long-acting testosterone undecanoate (TU) 1000 mg intramuscular injection. Treatment was initiated at day 1 (T1), then the second dose was administered 6 weeks later (T2). TU was injected at 3 months intervals thereafter. The parameters weight and waist circumference were measured at baseline and at every treatment visit. Serial BMI, WC and percentage of weight change from baseline were calculated.[br][bold]Results[/bold]: Median follow-up time was 4.7 years. The mean weight decrease from baseline to the last visit was 14.3 [plusmn] 8.7 kg (minimum -5, maximum 44), the mean percentage weight loss was 13.0 [plusmn] 6.8% (minumim -5.3, maximum 34.4), the mean of BMI points decreased was 4.5 [plusmn] 2.7 (minimum -1.6, maximum 13.4), and the mean drop in WC was 11 cm [plusmn] 6 (minimum -13, maximum 24). There is a strong linear relationship between weight loss, decrease in BMI and decrease in WC and time on TU treatment.[br][bold]Conclusion[/bold]: Long-term testosterone treatment with TU results in significant reduction of all parameters of obesity.[br][br]Sources of Research Support: Data entry was supported by Bayer Pharma AG.[br][br]Disclosures: AY: Speaker, Bayer Schering Pharma. RS: Speaker, Lilly USA, LLC. FS: Employee, Bayer Schering Pharma. Nothing to Disclose: YED, AY 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 477 53 229 SAT-116 PO28-02 Saturday 168 2012


169 ENDO12L_SAT-117 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) 15 Years of Experience with Intramuscular Testosterone Undecanoate for Substitution in Male Hypogonadism [mdash] Beneficial Effects on the Metabolic Syndrome and High Safety Profile Michael Zitzmann, Farid Saad University Clinics, Muenster, Germany; Gulf Medical University, Ajman, United Arab Emirates; Bayer Health Care, Berlin, Germany [bold]Background:[/bold][br]A reliable form of androgen substitution therapy in terms of favorable kinetics and tolerance as well as effective restoration of androgenicity is paramount for hypogonadal men. The intramuscular injection of the long-acting ester testosterone undecanoate (TU) offers a convenient modality for testosterone substitution.[br][bold]Methods:[/bold][br]We report data from 334 patients (147 with primary, 100 with secondary hypogonadism and 87 with late-onset ([ldquo]mixed[rdquo] or [ldquo]metabolic[rdquo] hypogonadism) aged 15 to 72 years (mean 42[plusmn]15 years) receiving altogether 6596 intramuscular injections of 1000 mg of TU during a maximal treatment time of 15 years, overall corresponding to 1403 treatment years.[br]Components of the metabolic syndrome were assesed in 269 men receiving 4296 injections.[br][bold]Results:[/bold][br]Individual dosing intervals ranged from 10 to 14 weeks Serum T concentrations increased from 5.8 to stable 16.1 nmol/L within the first year of treatment. The proportion of men fulfilling the new Harmonized Criteria for definition of the Metabolic Syndrome decreased from initially 88% to 52% within 2 years (Chi-square for trend: p[lt]0.001). During the maximal duration of treatment, an overall favorable change from baseline was visible for a multitude of parameters related to androgen effects/metabolic risk (see Table). Prostate size increased from 16.1[plusmn]5.2 to 21.1[plusmn]5.2 ml (p[lt]0.001), whilst PSA levels increased moderately (1.8[plusmn]0.4 to 1.9[plusmn]0.4[micro]g/l, p=0.001). No case of prostate cancer was observed. Hematocrit was significantly elevated during treatment but remained within the normal range (40.9[plusmn]2.1 to 46.2[plusmn]2.5%, p[lt]0.001). One patient suffered from deep vein thrombosis, one from stroke. No case of prostate cancer was observed.[br][bold]Parameter [rarr] Unit [rarr] Baseline [rarr] Endpoint 15 years [rarr] p[/bold] (ANOVA)[br]BMI [bold][rarr][/bold] kg[sup]x[/sup]m[sup]-2[/sup] [bold][rarr][/bold] 31.8[plusmn]5.2 [bold][rarr] [/bold]24.4[plusmn]3.2 [bold][rarr] [/bold][lt]0.001[br]Waist circ. [bold][rarr] [/bold]cm [bold][rarr] [/bold]114[plusmn]10.5 [bold][rarr] [/bold]94.1[plusmn]8.7 [bold][rarr] [/bold][lt]0.001[br]Weight [bold][rarr] [/bold]kg [bold][rarr] [/bold]103.0[plusmn]16.3 [bold][rarr] [/bold]79.1[plusmn]12.6 [bold][rarr] [/bold][lt]0.001[br]LDL-Cholesterol [bold][rarr][/bold] mg/dl [bold][rarr] [/bold]157[plusmn]29 [bold][rarr] [/bold]110[plusmn]19 [bold][rarr] [/bold][lt]0.001[br]HDL-Cholesterol [bold][rarr] [/bold]mg/dl [bold][rarr] [/bold]38.4[plusmn]9.7 [bold][rarr] [/bold]53.6[plusmn]11.7 [bold][rarr] [/bold][lt]0.001[br]Triglycerides [bold][rarr] [/bold]mg/dl [bold][rarr] [/bold]198[plusmn]33 [bold][rarr] [/bold]145[plusmn]21 [bold][rarr] [/bold][lt]0.001[br]Fasting glucose [bold][rarr] [/bold]mg/dl [bold][rarr] [/bold]118.1[plusmn]29.7 [bold][rarr] [/bold]91.2[plusmn]15.2 [bold][rarr] [/bold][lt]0.001[br]RR systolic [bold][rarr] [/bold]mmHg [bold][rarr] [/bold]148[plusmn]14 [bold][rarr] [/bold]128[plusmn]11 [bold][rarr] [/bold][lt]0.001[br]RR diastolic [bold][rarr] [/bold]mmHg [bold][rarr] [/bold]98[plusmn]11 [bold][rarr] [/bold]81[plusmn]10 [bold][rarr] [/bold][lt]0.001[br]Pulse [bold][rarr] [/bold]bpm [bold][rarr] [/bold]89[plusmn]9 [bold][rarr] [/bold]75[plusmn]8 [bold][rarr] [/bold][lt]0.001[br][bold]Conclusion:[/bold][br]Intramuscular injections of TU represent a feasible, safe and well tolerated modality of androgen substitution in hypogonadal men, substantiated by 15 years of experience, facilitating a decrement of metabolic/cardiovascular risk factors.[br][br]Disclosures: FS: Employee, Bayer Schering Pharma. Nothing to Disclose: MZ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1470 53 230 SAT-117 PO28-02 Saturday 169 2012


170 ENDO12L_SAT-118 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Restoring Testosterone to Normal Levels in Elderly Men Is Efficacious in Weight Reduction. A Follow-Up Study over 5 Years Farid Saad, Ahmad Haider, Louis Gooren Bayer Pharma AG, Berlin, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates; Private Practice, Bermerhaven, Germany; VUMC Amsterdam, Amsterdam, Netherlands [bold]Introduction:[/bold] Obesity is associated with reduced testosterone, and low testosterone induces weight gain. This study analysed the effects of normalization of serum testosterone in mainly elderly, hypogonadal men.[br][bold]Methods:[/bold] Open-label, single-center, cumulative, prospective registry study of 255 men (aged 38 [ndash] 83 years, mean 60.6 [plusmn] 8.0 years), with testosterone levels between 1.7 [ndash]3.5 ng/mL (mean: 2.87 [plusmn] 0.4). Cut-off point for testosterone treatment was serum testosterone [le] 3.5. ng/mL). 215 men were studied for at least 2 years, 182 for 3 years, 148 for 4 and 116 for at least 5 years. They received parenteral testosterone undecanoate 1000 mg/12 weeks after an initial interval of 6 weeks.[br][bold]Results:[/bold] After 5 years the following changes were observed: weight (kg) decreased from 106.22 [plusmn] 16.93 (minimum: 70, maximum: 139) to 90.07 [plusmn] 9.51 (min 74.00, max 115). The statistical significance was p[lt]0.0001 vs baseline and vs the previous year over 5 years indicating a continuous weight loss over the full observation period. Waist circumference (cm) declined from 107.24 [plusmn] 9.14 (min 86, max 129) to 98.46 [plusmn] 7.39 (min 84, max 117) (p[lt]0.0001 vs baseline and vs the previous year over 5 years). Body mass index (BMI, m/kg[sup]2[/sup]) declined from 33.93 [plusmn] 5.54 (min 21.91, max 46.51) to 29.17 [plusmn] 3.09 (min 22.7; max 36.71) (p[lt]0.0001 vs baseline and vs the previous year over 5 years). The mean per cent weight loss after 1 year was 4.12 [plusmn] 3.48%, after 2 years 7.47 [plusmn] 5.01%, after 3 years 9.01 [plusmn] 6.5%, after 4 years 11.26 [plusmn] 6.76% and after 5 years 13.21 [plusmn] 7.24%. At baseline, 96% of men had a waist circumference of [ge] 94 cm. This proportion decreased to 71% after 5 years.[br][bold]Conclusions:[/bold] Raising serum testosterone to normal produced loss of body weight, waist circumference and BMI. These improvements were progressive over the full 5 years of the study.[br][br]Sources of Research Support: Data entry and statistical analysis were supported by Bayer Pharma AG.[br][br]Disclosures: FS: Employee, Bayer Schering Pharma. AH: Principal Investigator, Bayer Schering Pharma. LG: Speaker, Bayer Schering Pharma. 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 495 53 231 SAT-118 PO28-02 Saturday 170 2012


171 ENDO12L_SAT-119 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Relationship between Cortisol and Components of the Metabolic Syndrome in Obese Children and Adolescents Chiara Guzzetti, Patrizia Zavattari, Sabrina Pilia, Anastasia Ibba, Maria Rosaria Casini, Luigi Minerba, Sandro Loche Ospedale Microcitemico, Cagliari, Italy; Universita[apos] degli Studi di Cagliari, Cagliari, Italy; Universita[apos] degli Studi di Cagliari, Cagliari, Italy [bold]Background.[/bold] A relationship between serum cortisol (F) and insulin resistance has been shown in obese adults and children, suggesting a role for F in the development of the metabolic syndrome (MS). In this regard, F could be an important predictor factor for MS.[br][bold]Objective.[/bold] The aim of this study was to evaluate the relationship between F and components of MS in obese children and adolescents.[br][bold]Subjects and Methods.[/bold] This is a retrospective study in 1027 obese children and adolescents (BMI-SDS 2.62[plusmn]0.51) referred to our endocrine unit in the last 8 y. Waist circumference (WC), systolic and diastolic blood pressure (SP, DP), morning serum concentrations of F, glucose (Glyc), cholesterol HDL, tryglicerides (TG) and HOMA (Glyc(mmol/L)xInsulin(mU/L)/22.5) were evaluated in all subjects. MS was defined according to the IDF criteria (1). Patients were subdivided into 3 age groups: 6-10, 10-16 and [gt]16 y.[br][bold]Results.[/bold] In univariate regression analysis including the entire cohort, F was weakly associated with SP (r=0.17, P[lt]0.001), DP (r=0.13 P[lt]0.001), Glyc (r=0.16 P[lt]0.001) and HOMA (r=0.15, P[lt]10[sup]-5[/sup]), also when adjusted for age, gender, puberty and BMI-SDS. 14/372 patients aged 10-16 y had MS. Univariate regression analysis in these 372 subjects showed that F was associated with SP (r=0.11, P=0.047), DP (r=0.16 P=0.003) and Glyc (r=0.21 P[lt]0.0001) but not with HOMA. When adjusted for age, gender, puberty and BMI-SDS, F was no longer correlated with SP but was weakly correlated with HOMA (r=0.12, P=0.02). In multivariate regression analysis including age, sex, puberty, BMI-SDS and F as independent variables and one of the component of the MS as the dependent variable, the only models where F was a weak predictor of the variability were one with DP (B=0.02, P=0.009, adjusted R[sup]2[/sup]=0.13) and one with Glyc (B=0.03, P=0.003, adjusted R[sup]2[/sup]=0.12) as dependent variables. Patients were then subdivided into 4 groups according to the WC centile: WC[lt]90, WC[gt]90, WC[gt]90 and 1 of the other components of the MS, WC[gt]90 and 2 of the other components of the MS. Mean F concentrations in the 4 groups was significantly different (F=7.12, P=0.0001), even adjusted for age, gender, puberty and BMI-SDS (F=4.53, P=0.004). Patients with one or more components of the MS had higher F concentrations.[br][bold]Conclusions.[/bold] In this cohort of obese children and adolescents, F was weakly associated with components of the MS. These findings suggest that F has a minor role, if any, in the development of MS.[br][br](1)Zimmet et al., Pediatric Diabetes 2007; 8:299[ndash]306.[br][br]Nothing to Disclose: CG, PZ, SP, AI, MRC, LM, SL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 478 53 232 SAT-119 PO28-02 Saturday 171 2012


172 ENDO12L_SAT-120 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Body Fat Is Inversely Correlated with Urine Free Cortisol in Human Subjects Nicola M Neary, Brent S Abel, Xiongce Zhao, Michael S Ring, Gail R Hall, Rana Malek, Kong Y Chen, Lynnette K Nieman, Monica C Skarulis National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; University of Maryland School of Medicine, Baltimore, MD [bold]Introduction: [/bold]Patients with anorexia nervosa are known to have increased plasma and urine free cortisol (UFC). We recently reported that UFC was elevated over 4-fold in a male with low body fat (1). In these situations, elevated cortisol may protect against further loss of visceral fat. We postulated that cortisol plays a role in fat regulation over a wider range of adiposities and hypothesized that body fat is inversely related to UFC in adults. The relationships between body fat as a percent of body weight (BF%) and AM and PM cortisol were also examined.[br][bold]Methods:[/bold] BF% was measured by DEXA in volunteers enrolled in a longitudinal study of overweight and obese adults. Inpatient assessment of the HPA axis included 24h UFC and mean diurnal serum cortisol obtained at 07:30, 08:00 and 23:30, 23:59. Patients treated with exogenous steroids, including creams and inhalers, or with diagnoses or treatment of depression, diabetes or obstructive sleep apnea or weight instability were excluded. For statistical analysis, log serum cortisol and square root UFC were used.[br][bold]Results: [/bold]212 subjects (89 males, 113 females) aged 18-69 (median 40, IQR 28-52) years, BMI 18-52 (median 28.7, IQR 24.1-34.7) kg/m[sup]2[/sup], of white (n=106, 50%), African American (n=83, 39%) and other (n=23, 11%) racial origin participated. UFC was median 21 (IQR 14-34) [mu]g/24h [normal range (nr) 3-45], AM cortisol: 10.9 (IQR 8.8-14.1) [mu]g/dl (nr 5-25), PM cortisol 2.8 (IQR 2.1-4.3), and BF% 35.4 (IQR 27.2-44.7). In univariate analysis, there was a weak but significant inverse correlation between BF% and UFC, R[sup]2[/sup]=0.08, p[lt]0.0002. After adjusting for race, sex, and age, BF% was inversely associated with AM cortisol (p=0.0321), but not with PM cortisol or UFC. Females had higher BF% than males (40.2[plusmn]0.9 (mean[plusmn]SEM) vs. 28.4[plusmn]1.0; p[lt]0.0001) but lower UFC (22.1[plusmn]1.4.2 vs. 29.7[plusmn]1.8, p=0.0002). Similarly, African American subjects had higher BF% than white subjects (37.8[plusmn]1.2 vs. 32.7[plusmn]1.1; p=0.0062) but lower UFC (20.9[plusmn]1.4 vs. 29.5[plusmn]1.9; p=0.0010).[br][bold]Summary: [/bold]In univariate analysis, BF% was weakly inversely correlated with UFC Adjusting for age, sex and race, BF% was inversely correlated with AM serum cortisol. There were significant differences in UFC between sexes and between racial groups that could relate in part to differences in body fat. Thus glucocorticoids may play a physiological role in fat preservation when the HPA axis is intact in addition to their critically adverse effect in excess as seen in Cushing[apos]s syndrome.[br][br](1)Neary NM et al., ENDO 2011; P3-226.[br][br]Sources of Research Support: National Institute of Diabetes and Digestive and Kidney Diseases; The Eunice Kennedy Shriver National Institute of Child Health and Human Development.[br][br]Nothing to Disclose: NMN, BSA, XZ, MSR, GRH, RM, KYC, LKN, MCS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1576 53 233 SAT-120 PO28-02 Saturday 172 2012


173 ENDO12L_SAT-121 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Estradiol Levels in Morbidly Obese Patients: Relationship with Metabolic and Inflammatory Markers Diana Teixeira, Diogo Pestana, Carla Sa, Sonia Norberto, Manuela Meireles, Gil Faria, Paula Freitas, Conceicao Calhau, Rosario Monteiro Faculty of Medicine, University of Porto, Porto, Portugal; S Jo[atilde]o Hospital, Faculty of Medicine, University of Porto, Porto, Portugal; S Jo[atilde]o Hospital, Faculty of Medicine, University of Porto, Porto, Portugal Obesity-related inflammation is a subject of great interest, not only for the exponential growth of incidence this pathology but also because obesity-driven inflammation may be an important instigator of the metabolic abnormalities that accompany the obese state. The dysregulation of estrogen metabolism has been associated with the susceptibility to obesity. However, the influence of estrogens on obesity-related inflammation has been little explored.[br]The aim of this study was to evaluate the relationship between total estradiol and inflammatory and metabolic markers in a sample of obese (BMI [gt] 40) patients undergoing bariatric surgery.[br]Twenty six men (age: 42.9 [plusmn] 9.3; BMI 44.6 [plusmn] 9.3; estradiol: 46.5 [plusmn] 33.9) and 221 women (age: 41.8 [plusmn] 10.6; BMI: 44.6 [plusmn] 4.9; estradiol 57.33 [plusmn] 63.90) divided by menopausal status (90 premenopausal: age: 38.0 [plusmn] 8.9; BMI 44.6 [plusmn] 4.3; estradiol: 66.6 [plusmn] 68.9; and 41 postmenopausal: age: 51.3 [plusmn] 8.7; BMI 44.9 [plusmn] 5.3; estradiol: 36.1 [plusmn] 40.8).[br]Body mass index (BMI), waist and hip circumference, blood pressure, lipid profile, plasma glucose, insulin, HgbA1c, estradiol, morning cortisol, sex hormone binding globulin (SHBG), testosterone, and hs-CPR were measured during standard clinical evaluation. Adipose tissue (subcutaneous and visceral) samples were collected during surgery at the Hospital of S. Jo[atilde]o, Portugal (protocol approved by the Ethics Committee of hospital) to determine size and number of adipocytes.[br]Plasma estradiol levels were positively correlated with BMI and HgbA1c in men and negatively correlated with morning cortisol levels. When all women were analyzed, estradiol levels were negatively correlated with: age, SHBG, testosterone and adipocyte number in visceral adipose tissue. In premenopausal women, estradiol levels were negatively associated with BMI, hip circumference, SHBG and positively correlated with alteration in HDL ([gt] 50 mg/dL) and IR-HOMA. There are no such significant correlations in postmenopausal women. No significant associations were found between estradiol and hs-CRP.[br]In conclusion, estradiol levels in men seem to be a marker of adiposity, and in women, seem to predict metabolic dysfunction, at least when all premenopausal women are analyzed as a whole.[br][br]Sources of Research Support: This work was supported by FCT (POCI, FEDER, Programa Comunit[aacute]rio de Apoio, PTDC/QUI/65501/2006; SFRH/BD/64691/2009; SFRH/BD/46640/2008, SFRH/BD/78367/2011,and SFRH/BPD/40110/2007).[br][br]Nothing to Disclose: DT, DP, CS, SN, MM, GF, PF, CC, RM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1446 53 234 SAT-121 PO28-02 Saturday 173 2012


174 ENDO12L_SAT-122 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Effects of GH Withdrawal in Obese Premenopausal Women with Abdominal Adiposity Eleanor Lin, Miriam A Bredella, Anu V Gerweck, Andrea L Utz, Karen K Miller Massachusetts General Hospital, Boston, MA; Vanderbilt, Nashville, TN [bold]Context[/bold]: Abdominal adiposity is associated with decreased growth hormone (GH) secretion. We previously reported results from a 6-month randomized, placebo-controlled study, which demonstrated that GH replacement in abdominally obese premenopausal women decreased total abdominal adipose tissue (TAT), subcutaneous abdominal adipose tissue (SAT), trunk fat, tissue plasminogen activator (tPA), apolipoprotein B (apoB), low-density lipoprotein (LDL), and high-sensitivity C-reactive protein (hsCRP), and increased thigh muscle and total lean mass. This was accompanied by a mild increase in both fasting and 2-hour glucose after oral glucose tolerance test (OGTT).[br][bold]Objective[/bold]: To determine whether GH replacement for 6 months has long-term effects on cardiovascular risk biomarkers, body composition, or glucose tolerance in obese women.[br][bold]Design[/bold]: Seventy-nine abdominally obese premenopausal women were allocated to GH (Genentech, Inc.) or placebo for 6 months. Thirty-nine completed a subsequent 6-month withdrawal observation period.[br][bold]Results[/bold]: Mean age (37[plusmn]1y) and BMI (34.9[plusmn]0.9kg/m[sup]2[/sup]) were comparable between the 39 subjects who completed the 6-month withdrawal period and the larger group that was initially randomized to GH or placebo. Insulin-like growth factor 1 (IGF-1) standard deviation scores (SDS) within the GH group were -1.7[plusmn]0.1 (pretreatment), -0.1[plusmn]0.3 (after 6 mos of GH) and -1.7[plusmn]0.1 (6 mos post GH withdrawal). Six months after GH withdrawal, TAT, SAT, total fat, trunk fat, trunk/extremity fat, hsCRP, apoB, LDL, and tPA increased ([italic]p[/italic][lt]0.05) by repeated-measures analysis of variance. There was a trend toward an increase in total cholesterol and extremity fat and a decrease in 2-hour OGTT glucose ([italic]p[/italic][lt]0.10). All body composition and cardiovascular risk marker outcome measures that had improved with GH returned to baseline levels by 6 months after GH discontinuation ([italic]p[/italic][gt]0.05). Fasting and 2-hour OGTT glucose returned to pre-treatment levels.[br][bold]Conclusion[/bold]: The effects of GH administration to abdominally obese premenopausal women have a short time-course. The beneficial effects on body composition and cardiovascular risk outcome measures, and the side effect of altered glucose tolerance, reverse after GH withdrawal; all return to pretreatment levels. Importantly, there was no indication of suppression of endogenous GH levels, as IGF-1 levels returned to baseline after withdrawal of GH therapy.[br][br]Sources of Research Support: This work was supported in part by National Institutes of Health Grants R01 HL-077674, UL1 RR-025758, K23 RR-23090, and T32 DK007028. Study medication and placebo only were supplied by Genentech, Inc.[br][br]Nothing to Disclose: EL, MAB, AVG, ALU, KKM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 679 53 235 SAT-122 PO28-02 Saturday 174 2012


175 ENDO12L_SAT-123 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Longitudinal Changes in Body Mass Index and Body Composition among Adult Survivors of Childhood Cancer Karin Blijdorp, Marry M van den Heuvel-Eibrink, Rob Pieters, AJ van der Lely, Sebastian JCMM Neggers Erasmus University Medical Centre Roterdam, Roterdam, Netherlands; Sophia Children[apos]s Hospital/Erasmus University Medical Centre Roterdam, Rotterdam, Netherlands Objective[br]Obesity, represented by high body mass index (BMI), is a major complication after treatment for childhood cancer. High amount of total and visceral fat and low lean body mass are described as more reliable determinants, predicting the development of cardiovascular disease. In this study longitudinal changes of BMI and body composition in adult childhood cancer survivors were evaluated.[br]Design Retrospective single center cohort study, evaluating longitudinal changes in BMI and body composition.[br]Subjects Data of 417 adult childhood cancer survivors, who had visited the late effects clinic twice, were analyzed. Median follow up time was 16 years (interquartile range 11-21) and time between visits was 3.2 (2.9-3.6). At both time points BMI was measured and body composition was assessed by dual X-ray absorptiometry (Lunar Prodigy). BMI and body composition measures were compared with those of healthy Dutch references and calculated as standard deviation scores (SDS).[br]Growth hormone deficient subjects treated with growth hormone replacement at time of follow up were excluded from further analyses.[br]Results[br]BMI SDS at 1st assessment was only significantly higher in female cranial radiotherapy (CRT) survivors as compared to healthy Dutch references (SDS=0.40, p=0.02). Increase of BMI over time (expressed as units per year) was only significantly higher in male survivors (0.27 versus 0.02 in controls (p[lt]0.001)). Percentage fat was significantly higher than controls in both men (SDS 1.37, P[lt]0.001) and women (SDS 1.05, P[lt]0.001) in all therapy groups, with the highest SDS after CRT (mean SDS 1.73 in men, 1.48 in women, P[lt]0.001). Only in men, increase in total fat percentage was significantly higher as compared to controls ([Delta]SDS=0.22, P[lt]0.001). Lean body mass did not significantly change over time.[br]Conclusion[br]Significantly greater increase of BMI and total fat percentage as compared to healthy references was found, especially in adult male survivors of childhood cancer.[br][br]Nothing to Disclose: KB, MMvdH-E, RP, AJvdL, SJCMMN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 627 53 236 SAT-123 PO28-02 Saturday 175 2012


176 ENDO12L_SAT-124 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Comparison of Fat Mass Index and Percent Body Fat as Measures of Adiposity in Youth David R Weber, Renee H Moore, Mary B Leonard, Babette S Zemel Children[apos]s Hospital of Philadelphia, Philadelphia, PA; University of Pennsylvania, Philadelphia, PA; Children[apos]s Hospital of Philadelphia, Philadelphia, PA; Children[apos]s Hospital of Philadelphia, Philadelphia, PA Background: Body Mass Index (BMI) of [ge]85th percentile is widely used to identify individuals at risk for excess adiposity and metabolic disease; however the ability of BMI to measure adiposity in youth may differ by racial group. Recent studies have suggested percentage body fat (PBF) as a superior index of adiposity in youth. The use of PBF as the gold standard of adiposity is an incomplete solution because it fails to account for the effect of height (HT). We hypothesize that fat mass index (fat mass/ht[sup]2[/sup], FMI) will more accurately represent adiposity in youth.[br]Methods: The LMS method was used to generate reference curves and Z-scores for FMI using total fat mass obtained from dual energy x-ray absorptiometry(DXA) scans in 8983 (3775 F) subjects aged 8-25 in 1999-2004 as a part of the National Health and Nutrition and Examination Survey. Comparisons of BMI and PBF were performed in 7123 (2903 F) subjects aged 8-20. Age and sex specific Z-scores for BMI (BMIZ) and HT (HTZ) were calculated using 2000 CDC growth reference data. PBF percentiles were defined using published reference data(1). Excess adiposity was defined as FMI or PBF [ge]75th percentile. Two-sample t-tests were used to compare continuous variables; chi-square tests for proportions.[br]Results: The positive predictive value (PPV) of BMI [ge]85th percentile to identify adiposity defined by FMI was lower in blacks (M:36%, F:30%) compared to whites (M:65%, F:52%), and Mexican Americans (M:73%, F:68%); (p[lt]0.001). Among subjects differentially classified by FMI and PBF, mean HTZ (M:0.71, F:0.48), BMIZ (M:1.54, F:1.62) and proportion black (M:17%, F:33%) were higher in subjects identified as having excess adiposity by FMI but normal by PBF, compared to HTZ (M:-0.04, F:-0.24), BMIZ(M:0.54, F:0.73) and proportion black (M:5%, F:5%) in subjects identified as having excess adiposity by PBF but normal by FMI (p[lt]0.02).[br]Conclusion: In a representative cohort of youth, the PPV of BMI [ge]85th percentile to identify excess adiposity defined by FMI is markedly lower in blacks. Subjects classified as having excess adiposity by FMI but normal adiposity by PBF are taller, have higher BMI, and are more likely to be black. We hypothesize that this represents a population of youth with high lean mass and high fat mass who may be at risk of metabolic disease but would be missed if using PBF to identify excess adiposity. Further studies are planned to investigate the effects of lean and fat mass on metabolic risk.[br][br](1)Ogden CL, Li Y, Freedman DS, Borrud LG, Flegal KM, Smoothed percentage body fat percentiles for US children and adolescents, 1999-2004. Natl Health Stat Report 2011;43:1-7.[br][br]Nothing to Disclose: DRW, RHM, MBL, BSZ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 632 53 237 SAT-124 PO28-02 Saturday 176 2012


177 ENDO12L_SAT-125 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Proteomic Identification of Morbidities in Overweight and Obese Prepubertal Children Nabila Roohi, Ateek Ahmad University of the Punjab, Lahore, Pakistan; Lahore General Hospital, Lahore, Pakistan [bold]Background and aims: [/bold]Childhood obesity is associated with significant morbidity and mortality in adult life. Proteomic identification of plasma signatures may be a valid tool for predicting and tracking the progression of comorbidities in overweight/obese children.[br][bold]Subjects and Methods:[/bold] We assessed plasma proteome of prepubertal boys (Mean age: 10.76[plusmn]0.23 years) with BMI 5[sup]th [/sup]to [lt]85[sup]th[/sup] percentile (healthy weight; n=12), 85[sup]th [/sup]to [lt]95[sup]th[/sup] percentile (overweight; n=20) and [ge]95[sup]th[/sup] percentile (obese; n=20) by two dimensional gel electrophoresis using Protean IEF coupled with II XL Cell System. The resolved proteins were identified by Swiss 2DPAGE database.[br][bold]Results: [/bold]By comparative proteome analysis, eight spots; five up-regulated and three down-regulated were differentially expressed in overweight and obese children in comparison to their normal weight counterparts. The expressions of Apolipoprotein (Apo) A-I, ApoA-IV and ApoE were significantly suppressed (p[lt]0.05) in 45%, 25% and 25% of overweight, whereas, 60%, 30% and 35% of obese children, respectively. ApoB-100, C-reactive protein (CRP), Retinol binding protein (RBP), Fibrinogen beta chain and Plasminogen activator inhibitor-1 (PAI-1) indicated marked expressions (p[lt]0.05) in 40%, 25%, 30%, 35% and 20% of overweight, while, 55%, 30%, 35%, 45% and 30% of obese children, respectively. The altered expressions of ApoA-I, ApoA-IV, ApoE, CRP, Fibrinogen and PAI-1 predict that overweight, in particular, obese children are at a higher risk of future progression towards cardiovascular disorders, particularly, atherosclerosis. Further, the over-expressions of CRP and RBP, in these children, demonstrae the risk for the development of insulin resistance and type 2 diabetes, in their later life.[br][bold]Conclusions: [/bold]Prominent alterations in the expressions of plasma proteins, in obese and overweight prepubertal children, may serve as novel diagnostic and prognostic biomarkers of morbidities in adulthood. Overweight, not only the obese, children should be screened for biomarkers. Extensive weight control strategies, for the prevention and reversal of overweight/obesity in children, may be warranted at population level.[br][br]Nothing to Disclose: NR, AA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 602 53 238 SAT-125 PO28-02 Saturday 177 2012


178 ENDO12L_SAT-126 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) High Second-Trimester Body Mass Index and Offspring Neurocognitive Development Wendy Y Craig, Glenn E Palomaki, Louis M Neveux, James E Haddow Foundation for Blood Research, Scarborough, ME; Women [amp] Infants Hospital/Alpert Medical School of Brown University, Providence, RI Among women and their offspring serving as controls for a separate nested case-control study, we observed an unexpected inverse association between second trimester maternal body mass index (BMI) and neurocognitive development at two years old. We explored this association further in that dataset, and in a second control group from an earlier nested case-control study that measured neurocognitive development in eight year old offspring. Both study groups were recruited from the Maine pregnancy population; demographic and pregnancy data were obtained at the time of prenatal screening and from the birth record. We tested 101 children (mothers pregnant 2004-6) using the Bayley Scales of Infant Development (BSID-III), and 118 children at age eight years (mothers pregnant 1987-90), using the Wechsler Intelligence Scale for Children (WISC-III). Frequency of maternal BMI [ge]30 kg/m[sup]2[/sup] increased from 10% to 30% in the 15 years between studies (p[lt]0.001), and the socioeconomic gradient with BMI became more pronounced (measured by the Hollingshead score at the time of neurocognitive testing). Regression analysis showed an inverse relationship between BSID-III language scores and log BMI in two year old offspring, after adjustment for potential effect modifiers (r = -0.16, slope = -26.810, p = 0.05; intercept = 138.8594); there was no significant relationship between BMI and either cognitive or motor scores. The inverse relationship between WISC-III performance subscale IQ scores and log BMI at eight years remained significant after adjustment (r = -0.20, slope = -34.873, p= 0.023; intercept = 148.534); there was no significant relationship between BMI and either full scale IQ or verbal subscale scores. Maternal obesity may be an independent risk factor for children[apos]s neurocognitive development. Confirmatory studies are needed.[br][br]Nothing to Disclose: WYC, GEP, LMN, JEH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 670 53 239 SAT-126 PO28-02 Saturday 178 2012


179 ENDO12L_SAT-127 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Relationship of Body Composition with Bone Mass in Korean Adults Kyung-Soo Kim, Soo-Kyung Kim, Se-Hwa Kim, Seok Won Park, Yong-Wook Cho CHA Bundang Medical Center, CHA University, Seongnam, Republic of Korea; Kwandong University College of Medicine, Goyang, Republic of Korea A relationship between bone mineral density (BMD) and body mass index or weight has been established. However, the effect of body composition such as fat or muscle mass on bone mass remains controversial. The aim of the present study was to examine the relationship of body composition with bone mass in Korean adults based on a large population-based survey.[br]Cross-sectional analysis were carried out on 3643 Korean adults (1928 men, 705 premenopausal women, 1010 postmenopausal women) who were 40 years or older and participated in the Fourth Korea National Health and Nutrition Examination Survey conducted in 2009. Total and regional fat mass, lean mass, and bone mineral content (BMC), BMD were measured by dual energy X-ray absorptiometry.[br]In normal (18.5[le]BMI[lt]23) and overweight (23[le]BMI[lt]25) group of premenopausal women, lumbar spine BMC was significantly higher in low waist circumference (WC) ([lt]80cm) than in high WC ([ge]80cm) after adjustment of age and weight (P[lt]0.05). Also in overweight group of postmenopausal women, lumbar spine BMC was significantly higher in low WC than in high WC after adjustment of age and weight (P[lt]0.05). In men, lumbar spine BMC was high in low WC in overweight group but in high WC in obese (BMI[ge]25) group. We showed both total fat mass and total lean mass were positively correlated with BMD (femur neck, lumbar spine) in all three groups. However, after controlling for age and weight, the association between total fat mass and BMD (femur neck, lumbar spine) became negative (P[lt]0.05). Furthermore, after controlling for age and weight, there were no significant association between appendicular fat mass and femur neck BMD although truncal fat mass still showed a negative association in premenopausal and postmenopausal women. In contrast, there was a positive association between total and regional lean mass (arm and leg) and BMD (femur neck and lumbar spine) after adjustment of age and weight in all three groups.[br]In conclusion, fat mass was negatively correlated with BMD (femur and lumbar spine) and lean mass was positively after removing the mechanical loading effect in Korean adults. Especially, in femur neck BMD, truncal fat mass was more important than appendicular fat mass in women.[br][br]Nothing to Disclose: K-SK, S-KK, S-HK, SWP, Y-WC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 688 53 240 SAT-127 PO28-02 Saturday 179 2012


180 ENDO12L_SAT-128 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Gender Difference in Secular Trends of Anthropometric Parameters among Korean Adolescents, 1998[sim]2008 Ki Eun Kim, Shin Hye Kim, Sang Shin Park, Mi Jung Park Gangnam CHA Hospital, CHA University, School of Medicine, Seoul, Republic of Korea; Sanggye Paik Hospital, Inje University, College of Medicine, Seuol, Republic of Korea; College of Veterinary Medicine and Biomedical Sciences, Texas A[amp]M University, College Station, Canyon, TX; Sanggye Paik Hospital, Inje University, College of Medicine, Seoul, Republic of Korea Objectives[br]The aim of this study was to investigate national trends of body physique [height, weight, body mass index (BMI), and waist circumference] in Korean adolescents.[br]Methods[br]We analyzed data for 2,822 subjects aged 10-18 yr (1,478 boys and 1,344 girls) from the Korean National Health and Nutrition Examination Surveys conducted between 1998 and 2008.[br]Results:[br]The final height of boys increased from 172.5 [plusmn] 6.6 cm in 1998 to 175.5 [plusmn] 5.1 cm in 2008 (P=0.008), while there was no significant change in the final height of girls. Over the study period, mean values of body weight, BMI and waist circumference were significantly increased in boys; however no significant changes were observed in girls. The proportion of boys with normal weight was decreased from 82.8% to 71.4% (P[lt]0.0001), while the proportion of overweight (from 6.3% to 14.7%, P[lt]0.0001) and obese boys (from 4.7% to 8.2%, P=0.004) were significantly increased. No significant difference in proportions of normal weight, overweight and obese girls was seen over the study period, however increasing trend in proportion of underweight girls was observed in elementary school girls (from 4.8% to 9.4%, P=0.05).[br]Conclusions[br]Secular trend of anthropometric parameters in Korean adolescents was marked by increasement of mean final height, obesity prevalence in teen aged boys, and emergence of underweight in elementary school girls. Age and gender targeted measures to prevent obesity and underweight are warranted in health policies for Korean youth.[br][br]Nothing to Disclose: KEK, SHK, SSP, MJP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 708 53 241 SAT-128 PO28-02 Saturday 180 2012


181 ENDO12L_SAT-129 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Normative Values of Anthropometric Indices among Residents of Calabar, South East Nigeria Ofem Egbe Enang, Okon Ekwere Essien, Olufemi Adetola Fasanmade, Augustine Efedaye Ohwovoriole University of Calabar Teaching Hospital, Calabar, Nigeria; Lagos University Teaching Hospital, Lagos, Nigeria Background: Ethnic specific anthropometric values have been proposed by the International Diabetes Federation (IDF) to aid in the diagnosis of the metabolic syndrome. No such values are available for Sub-Saharan Africa including Nigeria.[br]Objectives: To determine and propose normative values of indices of anthropometry among the inhabitants of a coastal Nigerian city.[br]Methods: In a cross sectional survey, a sample comprising 1134 subjects (645 males and 489 females) representative of the entire population of Calabar aged 15[ndash]79 was studied. A multistage sampling method was applied and 50 house-holds from each of the four wards were selected using the table of random numbers, out of which eligible individuals aged between 15 years and 79 years from the 200 households selected were recruited.Using a modification of WHO STEPS instrument the information obtained included anthropometric indices (height in meters, weight in kilogram, waist circumference in centimetre, Hip circumference in centimetre). Anthropometric indices were expressed as mean (standard deviation). The normative values of indices of nutriture were determined using mean at 95% confidence interval, and the level of significance was taken at P [lt] 0.05.[br]Results: The mean (95% CI) values of BMI for males and females were 27.0kg/m2 (95% CI 26.5-27.2) and 28.5kg/m2 (95% CI 28.0-29.0) respectively, P [lt]0.01. The mean (95% CI) value of WC males was 91cm (95% CI 90.2-91.8), of females was 89.8cm (95% CI 88.8-90.8), P [lt] 0.01. The mean (95% CI) values of WHR males was 0.90 (95% CI 0.88-0.89) while that of females was 0.85 (95% CI 0.87-0.89) P [lt] 0.01. The mean (95% CI) values of height of males was 1.70m (1.70-1.72) and of females was 1.60m (1.64-1.65), P [lt]0.01.[br]Conclusion: The normative values of anthropometry in the study population are different from those from other parts of Nigeria and other parts of the world with lower BMI, higher WC and WHR.More studies on anthropometric indices, involving other regions of the country are needed, to aid in the establishment of nationwide specific cut-off values for Nigeria.[br][br]Nothing to Disclose: OEE, OEE, OAF, AEO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 315 53 242 SAT-129 PO28-02 Saturday 181 2012


182 ENDO12L_SAT-130 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) IGF-1, IGFBP-1, IGFBP-3 and IGFBP-4 Levels and Their Associations to Cardiovascular Risk Factors in Obese Children before and after Weight Loss Thomas Reinehr, Joachim Woelfle, Nina Lass, Yvonne Uysal, Christian Roth Vestische Children[apos]Hospital, University of Witten/Herdecke, Datteln, Germany; University of Bonn, Bonn, Germany; University of Washington, Seattle, WA [bold]Background[/bold]: Insulin-like growth factor (IGF) -1 and its binding proteins (IGFBP-1,-3,-4) are postulated to be associated to obesity, insulin resistance and its associated comorbidities. However, studies in children are controversial and most importantly longitudinal studies are scarce.[br][bold]Methods: [/bold]We analyzed IGF-1, IGFBP-1, IGFBP-3, and IGFBP-4 in 60 obese children (mean age 10.8[plusmn]2.4, 52% female, mean BMI 28.2[plusmn]4.1kg/m[sup2]) at onset and end of a one-year lifestyle intervention. Anthropometrical markers, waist circumference (WC), systolic (SBP) and diastolic blood pressure (DBP), leptin, adiponectin, fasting glucose, insulin, triglycerides, uric acid, LDL- and HDL-cholesterol concentrations were determined at baseline and 1 year later.[br][bold]Results[/bold]: At baseline, IGF-1 correlated significantly to SBP (r=0.39), DBP (r=0.21), HDL-cholesterol (r=-0.30), and triglycerides (r=0.37). IGFBP-1 correlated significantly to BMI (r=-0.48), WC (r=-0.44), SBP (r=0.37), DBP (r=0.26), insulin (r=0.25), HOMA (r=-0.33), leptin (r=-0.37), and adiponectin (r=0.34). IGFBP-3 correlated significantly to WC (r=-0.39), SBP (r=-0.37), DBP (r=-0.30), HDL-cholesterol (r=-0.32), LDL-cholesterol (r=0.33), triglycerides (r=0.28), uric acid (r=0.25), insulin (r=0.25), HOMA (r=0.24), leptin (r=0.21), and adiponectin (r=-0.40). IGFBP-4 correlated significantly to BMI (r=0.34), WC (r=0.39), uric acid (r=0.39), insulin (r=0.32), HOMA (r=0.34), leptin (r=0.30), and adiponectin (r=-0.28).[br]In the course of 1y, the changes of IGF-1 were significantly related to changes of triglycerides (r=0.35) and uric acid (r=0.27). The changes of IGFBP-3 correlated significantly to changes of WC (r=0.29), SBP (r=0.29) and DBP (r=0.20). The changes of IGFBP-4 were significantly related to changes of BMI (r=0.21), uric acid (r=0.26), insulin (r=0.21), and HOMA (r=0.22).[br]In the 30 children with reduction of BMI (in mean -2.9kg/m[sup2]) all cardiovascular risk factors improved significantly, while IGF-1, IGFBP-3, and IGFBP-4 did not change significantly. In the 30 children with increase of BMI (in mean +1.3kg/m[sup2]), IGF-1 and IGFBP-4 increased significantly, IGFBP-1 decreased significantly, while IGFBP-3 and all cardiovascular risk factors remained stable.[br][bold]Conclusion[/bold]: The cross-sectional and longitudinal relationships between IGF-1 and its binding proteins to cardiovascular risk factors suggest a potential role of the IGF-1 axis in the pathogenesis of insulin resistance and its associated comorbidities.[br][br]Sources of Research Support: This work was supported by an investigator-initiated, unrestricted research grant from Merck Serono GmbH.[br][br]Nothing to Disclose: TR, JW, NL, YU, CR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 10 53 243 SAT-130 PO28-02 Saturday 182 2012


183 ENDO12L_SAT-131 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Waist Circumference Is Superior to Weight and BMI in Predicting Sexual Symptoms, Voiding Symptoms and Psychosomatic Symptoms in Men with Late-Onset Hypogonadism Aksam Yassin, Youssef El Douaihy, Ridwan Shabsigh, Aiman Yassin, Farid Saad Segeberger Kliniken, Norderstedt, Germany; Dresden International University, Dresden, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates; Maimonides Medical Center, Brooklyn, NY; Klinikum Braunschweig, Braunschweig, Germany; Bayer Pharma AG, Berlin, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates [bold]Introduction[/bold]: We investigated whether three measurements of obesity: weight, BMI, and waist circumference correlate with the International Index of Erectile Function (IIEF), the Aging Males[apos] Symptoms questionnaire (AMS), and the International Prostate Symptom Score (IPSS).[br][bold]Materials and methods[/bold]: As of November 2004, 130 patients were enrolled in a prospective study on late onset hypogonadism (LOH defined as total testosterone [le] 3.5 ng/mL and symptoms). All men were treated with i.m. testosterone undecanoate (TU) 1000mg. The treatment was initiated on day 1, then 6 weeks thereafter and was continued at 3 months intervals. The mean time of treatment and follow up was 4.7 years. Baseline demographic data were recorded including hormones, lipids, and glucose. Blood samples were collected at every or every other visit. In addition patients were asked to fill in three standardized questionnaires IIEF, IPSS and AMS. The Sexual Health Inventory for Men (SHIM), a short version of the IIEF was used. SPSS was used to plot weight, BMI and waist circumference versus the recorded scores on the three different questionnaires. ANOVA analysis was used to look at any possible statistical significance.[br][bold]Results[/bold]: The cohort[apos]s means and standard deviations for the IIEF, AMS and IPSS scores were 7.3 ([plusmn]3.11), 54.63 ([plusmn]7.06) and 11.65 ([plusmn]4.41) respectively. The estimated curves and the ANOVA regression for continuous variables showed that waist circumference is inversely proportional to IIEF and directly proportional to AMS and IPSS with statistical significance. Weight was inversely proportional to IIEF, and directly proportional to IPSS both with statistical significance. Weight did not show enough linearity with AMS to give statistical significance. BMI and all three measurements of size had no proportionality.[br][bold]Conclusion[/bold]: Among weight, BMI and waist circumference, the latter is the best predictor of health related quality of life in men with LOH including sexual symptoms, voiding symptoms and other psychosomatic symptoms.[br][br]Sources of Research Support: Data entry has been supported by Bayer Pharma AG.[br][br]Disclosures: AY: Speaker, Bayer Schering Pharma. RS: Speaker, Lilly USA, LLC. FS: Employee, Bayer Schering Pharma. Nothing to Disclose: YED, AY 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 469 53 244 SAT-131 PO28-02 Saturday 183 2012


184 ENDO12L_SAT-132 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Depressive Symptoms in Adolescents Referred for Obesity Management: What Is the Relationship with Insulin Resistance? Tamara S Hannon, Ann M Lagges, Jordan N Huber, Aaron E Carroll, Sandeep K Gupta Indiana University School of Medicine, Indianapolis, IN; Indiana University School of Medicine, Indianapolis, IN; Indiana University School of Medicine, Indianapolis, IN; Indiana University School of Medicine, Indianapolis, IN Studies in adults have shown depression and psychosocial stress to be associated with metabolic syndrome and type 2 diabetes; however, few studies in pediatrics have examined these relationships. We aimed to determine whether depressive symptoms are related to measures of glucose tolerance (fasting glucose, glycosylated hemoglobin [A1C]) and insulin resistance (fasting insulin, HOMA-IR) in obese adolescents referred to a hospital-based obesity program (Pediatric OverWeight Education and Research, POWER). Methods: The Children[apos]s Depression Inventory[sup]1[/sup] (CDI) was used to examine self-reported depressive symptoms (anhedonia, self-esteem, ineffectiveness, interpersonal problems, negative mood) in 284 children referred for treatment of obesity. Of the 284 patients, 260 had complete CDI and laboratory data (64% female; 54% white, 26% black, 5% Hispanic, 3% other race; mean age 12.9[plusmn]2.7 y, BMI 37.3[plusmn]8.1 kg/m[sup]2[/sup]). Results: BMI was associated with insulin resistance (r=0.37, p[lt]0.001 for fasting insulin; r=0.32, p[lt]0.001 for HOMA-IR) but not with glucose tolerance (r=[minus]0.09, p=0.20 for fasting glucose; r=0.08, p=0.21 for A1C). CDI t-scores (mean=49; range 31-100) were not indicative of increased prevalence of depression in this population and were not associated with BMI (r=0.04, p=0.54). CDI t-scores were, however, positively correlated with insulin resistance (r=0.15, p=0.02 for fasting insulin and r=0.16, p=0.11 for HOMA-IR), but not with glucose tolerance. These relationships were largely due to associations with interpersonal problems (difficulties interacting with people, social avoidance and isolation) (r=0.16, p=0.01 for fasting insulin; r=0.13, p=0.03 for fasting glucose; r=0.16, p=0.01 for HOMA-IR). Multiple linear regression analysis was performed controlling for sex, race, age, and BMI to determine the degree to which interpersonal problems predicted insulin resistance. The total variance explained by the model was 20%, F=10.4, p[lt]0.001. The interpersonal problem t-score contributed significantly to the model, R[sup]2[/sup] change=0.11, F change=17.5, p[lt]0.001. Conclusions: Among adolescents referred for obesity treatment, endorsement of interpersonal problems is associated with insulin resistance. The relationships between interpersonal problems and risk factors for the development of diabetes in youth should be investigated further to evaluate the mechanisms and determine if early recognition and therapy could lessen the risk of type 2 diabetes.[br][br](1) Kovacs, M. (2003) Children[apos]s Depression Inventory. Western Psychological Services, Los Angeles, CA.[br][br]Nothing to Disclose: TSH, AML, JNH, AEC, SKG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 943 53 245 SAT-132 PO28-02 Saturday 184 2012


185 ENDO12L_SAT-133 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Effects of Different Bariatric Surgery Procedures on Oligoelements and Antioxidants in Human Obesity Antonio Mancini, Roberto Festa, Sebastiano Raimondo, Chantal Di Segni, Serena Marchitelli, Andrea Silvestrini, Elisabetta Meucci, Alfredo Pontecorvi, Roberto Maria Tacchino Catholic University of the Sacred Heart, Rome, Italy; Polytechnic University of the Marche, Ancona, Italy; Catholic University of the Sacred Heart, Rome, Italy; Catholic University of the Sacred Heart, Rome, Italy In previous works we have demonstrated that surgically-induced weight loss, obtained by biliopancreatic diversion (BPD), is associated with an increase in insulin sensitivity, but a decrease in some molecules, such as Coenzyme Q[sub]10[/sub], that act as powerful antioxidant system. The mechanism of this phenomenon (weight loss per se, lipid malabsorption, metabolic or hormonal variations after surgery) are not yet clear. Since antioxidants are very important in preventing metabolic complications of obesity, in order to delve into the physiopathology of this condition we have studied 19 subjects (6 males and 13 females, age 41.4 [plusmn] 11.0 years, BMI 48.5 [plusmn] 7.9 Kg/m[sup]2[/sup]), before and at a long time (12-18 months) from surgery, treated by different surgical procedures: BPD, gastric bypass (GB), gastric minibypass (GM), evaluating oligoelements and antioxidants in plasma.[br]After surgery they reached a mean % decrease in body weight of 41.7 [plusmn] 0.1, without differences with the different surgical techniques. They were treated with the same schedule of integrative therapy. We studied metabolic parameters (glycemia, insulinemia with the calculation of HOMA insulin-resistance index, total- LDL- HDL-cholesterol, triglycerides, uric acid, albumin, transaminases), hormones (fT3, fT4, TSH, IGF-1, cortisol, ACTH) and serum values of copper, zinc and Vitamin E. Total antioxidant capacity was evaluated by the system metmyoglobin-H[sub]2[/sub]O[sub]2[/sub], as a source of radicals, and the chromogen ABTS, whose radical cation is spectroscopically revealed: the latency time (sec) in the appearance of radical species is a measurement of the antioxidant content of the sample. Analysis of variance was used for statistical evaluation.[br]We observed significant differences (p[lt]0.05) in mean variation of: HOMA (-80[plusmn]11% in BPD, -55[plusmn]31% in GB, -84[plusmn]15% in GM), HDL-cholesterol (+3[plusmn]21%, +26[plusmn]30%, +32[plusmn]17%, respectively), IGF-1 (-41[plusmn]21%, +9[plusmn]37%, -50[plusmn]32%, respectively). LAG values did not significantly differed in the three groups (-12[plusmn]14%, -18[plusmn]14%, -13[plusmn]21%, respectively).[br]These data, although preliminary, suggest a different pattern of response to bariatric surgery, irrespective of weight loss, with a more favourable response in gastric bypass and minibypass in comparison to BPD. They also suggest that previously demonstrated loss of antioxidant could be due to lipid malabsorption as main mechanism.[br][br]Nothing to Disclose: AM, RF, SR, CDS, SM, AS, EM, AP, RMT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 946 53 246 SAT-133 PO28-02 Saturday 185 2012


186 ENDO12L_SAT-134 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) How Does Treating Sleep-Disordered Breathing Affect the Lipid Profile in Obese Children and Adolescents? Zarlasht Amini, Seema Kumar, Suresh Kotagal, Robin Lloyd, Christine Lohse Mayo College of Medicine, Rochester, MN; Mayo College of Medicine, Rochester, MN; Mayo College of Medicine, Rochester, MN [bold]Background: [/bold]Childhood obesity is frequently associated with hyperlipidemia, hypertension, insulin resistance and cardiovascular risk. Obstructive Sleep Apnea (OSA) seen commonly in obese children appears to contribute to greater risk of these comorbidities independent of obesity. However the impact of treatment of OSA on metabolic parameters in children is not known. The objective of the study was to assess the impact of treating OSA with Positive Airway Pressure breathing (PAP) or Adenotonsillectomy (T[amp]A) on the fasting glucose and lipid profile in obese children and adolescents.[br][bold]Methods: [/bold]Retrospective medical record review of subjects aged 2-18 years with BMI[gt] 85[sup]th[/sup] percentile for age and gender who underwent polysomnography at the Center for Sleep Medicine for evaluation of Sleep Disordered Breathing (SDB) between January 1[sup]st[/sup], 2000 and December 31[sup]st[/sup], 2010. Associations among variables were evaluated using Spearman rank correlation coefficients and changes in variables from baseline following treatment were evaluated using paired t and Wilcoxon signed rank tests.[br][bold]Results: [/bold]Forty seven patients with OSA (defined as Apnea Hypopnea Index (AHI) of [ge] 1 per hour on overnight polysomnography) had a fasting lipid panel consisting of total cholesterol, triglycerides, high-density lipoprotein (HDL) cholesterol and calculated low density lipoprotein (LDL) cholesterol measured within three months prior to the diagnosis of OSA. Forty four patients had a fasting glucose measured within three months prior to the diagnosis of OSA. OSA was treated with PAP or T[amp]A. Twenty four subjects had a repeat lipid panel and 18 subjects had a repeat fasting glucose drawn at a mean of 19 months (median 12.5; range 3 [ndash] 46) following treatment of OSA.[br][bold]Results:[/bold] AHI was significantly positively correlated with fasting glucose (r= 0.45, p=0.002) and with systolic blood pressure (r=0.24, p=0.023). There was a statistically significant decrease in total cholesterol and in LDL cholesterol following treatment of OSA (median change -12.5 mg/dL; p[lt]0.001 and -4.5 mg/dL; p=0.021, respectively). No significant change in HDL-cholesterol, triglycerides or fasting glucose was noted.[br][bold]Conclusion[/bold]: Treatment of OSA in obese children is associated with improvement in levels of total cholesterol and LDL-cholesterol levels. Screening for OSA in obese children and its treatment should be an integral component of programs aimed at decreasing cardiovascular risk in these children.[br][br]Nothing to Disclose: ZA, SK, SK, RL, CL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 958 53 247 SAT-134 PO28-02 Saturday 186 2012


187 ENDO12L_SAT-135 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Effect of CPAP on Weight in Patients with Sleep Apnea: A Meta-Analysis Zunaira Z Chaudhry, Ali Hamza, Carmella Evans-Molina, Rashid Nadeem, Janice Gilden, Saleha Z Chaudhry, Abdul Rehman Rishi, Muhammad A Rishi Indiana University, Indianapolis, IN; Captain James A Lovell Federal Health Care Center, North Chicago, IL; Mercy Hospital Medical Center, Chicago, IL; MacNeal Hospital Medical Center, Berwyn, IL Background: Obesity is a common cause of obstructive sleep apnea syndrome (OSAS). Impact of treatment of sleep apnea with continuous positive airway pressure (CPAP) on body mass index (BMI) is not well established with various studies producing differing results.[br]Methods: We conducted a meta-analysis of controlled trials to evaluate the effects of CPAP on BMI, reported as either a primary or secondary end point, among patients with OSAS. Studies were retrieved by searching Medline (January 1980 to Nov 2011), the Cochrane Database of Systematic Reviews, conference abstracts, and bibliographies of review and original articles. From 345 relevant reports, 10 randomized control clinical trials were selected that compared CPAP to control among participants with OSAS, had a minimum treatment duration of 4 weeks, and reported BMI changes during the intervention or control period. Data on sample size, participant characteristics, study design, intervention methods, duration, and treatment results were independently abstracted by 2 investigators using a standardized protocol and examined using a fixed-effects model.[br]Results: Mean net change in BMI in patients with OSAS treated with CPAP compared with control was 0.78 kg/m[sup2] (95% confidence interval; CI 0.65-0.91).[br]Conclusion: This meta analysis indicates that CPAP use in patients with OSAS may lead to a modest increase in BMI. Further studies are needed to elucidate its cause.[br][br]Nothing to Disclose: ZZC, AH, CE-M, RN, JG, SZC, ARR, MAR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2358 53 248 SAT-135 PO28-02 Saturday 187 2012


188 ENDO12L_SAT-136 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Abdominal Adiposity and Obstructive Airway Disease: Testing Insulin Resistance and Sleep-Disordered Breathing Mechanisms Matthew Timothy Haren, Gary Misan, Tracey-Jayne Paterson, Richard E Ruffin, Janet F Grant, Jonathan D Buckley, Peter RC Howe, Jonathan Newbury, Anne W Taylor, Robyn A McDermott University of South Australia, Adelaide, Australia; University of South Australia and The University of Adelaide, Whyalla, Australia; University of South Australia, Whyalla, Australia; University of Adelaide, Adelaide, Australia; University of South Australia, Adelaide, Australia [bold]Background:[/bold] This study examined associations of abdominal adiposity with lung function, asthma symptoms and current doctor-diagnosed asthma, and mediation by insulin resistance (IR) and sleep disordered breathing (SDB).[br][bold]Methods:[/bold] A random sample of 2500 households was drawn from the community of Whyalla, South Australia (The Whyalla Intergenerational Study of Health, WISH February 2008 - July 2009). Seven-hundred twenty-two randomly selected adults (18 years and over) completed clinical protocols (32.2% response rate). Lung function was measured by spirometry. Post-bronchodilator FEV[sub]1[/sub]/FVC was used to measure airway obstruction, and reversibility of FEV[sub]1[/sub] was calculated. Current asthma was defined by self-reported doctor-diagnosis and evidence of currently active asthma. Symptom scores for asthma (CASS) and SDB were calculated. Intra-abdominal fat (IAF) was estimated using dual-energy x-ray absorptiometry (DXA). IR (HOMA2-IR) was calculated from fasting glucose and insulin concentrations.[br][bold]Results:[/bold] The prevalence of current doctor-diagnosed asthma was 19.9% (95% CI 16.7, 23.5%). The ratio of observed to expected cases given the age and sex distribution of the population was 2.4 (95%CI 2.1, 2.9). IAF was not associated with current doctor-diagnosed asthma, FEV[sub]1[/sub]/FVC or FEV[sub]1[/sub] reversibility in men or women but was positively associated with CASS independent of IR and SDB in women. A 1% increase in IAF was associated with decreases of 12mL and 20mL in FEV[sub]1[/sub] and FVC respectively in men, and 4mL and 7mL respectively in women. SDB mediated 12% and 26% of these associations respectively in men only.[br][bold]Conclusions:[/bold] In this population with an excess of doctor-diagnosed asthma, IAF was not a major factor in airway obstruction or doctor-diagnosed asthma, although women with higher IAF perceived more severe asthma symptoms which did not correlate with lower FEV[sub]1[/sub]. Higher IAF was significantly associated with lower FEV[sub]1[/sub] and FVC and in men SDB mechanisms may contribute up to one quarter of this association.[br][br]Sources of Research Support: The South Australian Premiers Science Research Fund; University Departments of Rural Health Program (Australian Government Department of Health and Ageing); National Health and Medical Research Council (NHMRC) of Australia Post-doctoral Training Fellowship (Public Health) 511345 awarded to MTH; NHMRC Public Health Practitioner Fellowship awarded to RAM.[br][br]Nothing to Disclose: MTH, GM, T-JP, RER, JFG, JDB, PRCH, JN, AWT, RAM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1315 53 249 SAT-136 PO28-02 Saturday 188 2012


189 ENDO12L_SAT-137 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) The Ability of Body Mass Index To Predict Abnormal Waist Circumference: ROC Analysis Marcio W Lauria, Livia P Moreira, George M Coelho, Silvia N Freitas, Raimundo Marques, Adauto V Ramos, Maria Marta S Soares UFMG, Belo Horizonte, Brazil; Hospital Felicio Rocho, Belo Horizonte, Brazil; UFOP, Ouro Preto, Brazil Background: Body mass index (BMI) is widely utilized as an anthropometric estimate of general adiposity; however, differences in body composition and body fat distribution limit its usefulness. In recognition that visceral fat accumulation increases the risk for metabolic disease, waist circumference (WC) was promoted as an alternative surrogate measure of obesity. However, WC is subject to significant inter-examiner variation.[br]Objectives: To correlate BMI and WC measures in a group of Brazilian adults searching for the most accurate BMI values in predicting abnormal WC.[br]Methods: BMI and WC were measured in 1,184 volunteers (45.6[plusmn]17.3 yrs; 69% women) according to standard procedures. Abnormal WC was defined as [gt] 88 cm in women and [gt] 102 cm in men. Statistics analysis was done using Pearson[acute]s correlation coefficient and receiver operating characteristic (ROC) curve.[br]Results: Anthropometric measures: BMI- women=27.1[plusmn]5.9 kg/m2; WC-women=88.9[plusmn]15.1 cm; BMI-men=25.4[plusmn]4.8 kg/m2; WC-men=89.9[plusmn]14.4 cm. BMI had a strong correlation with WC (women: r=0.87, p[lt]0.0001, area under ROC curve=0.93[plusmn]0.01; men: r=0.89, p[lt]0.0001, area under ROC curve=0.94[plusmn]0.01). The most accurate BMI cut offs for abnormal WC were 26.8kg/m2 for women (sensibility=82%; specificity=89%) and 27.1 kg/m2 for men (sensibility=96%; specificity=81%).[br]Conclusion: Patients with BMI[gt] 27 kg/m2 usually have abnormal WC. In this group, WC measurement may reasonably be waived. For patients with lower BMI, WC measurement still remains informative.[br][br]Nothing to Disclose: MWL, LPM, GMC, SNF, RM, AVR, MMSS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 995 53 250 SAT-137 PO28-02 Saturday 189 2012


190 ENDO12L_SAT-138 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Association between Mitochondrial DNA Content, Mitochondrial Function, and Childhood Obesity and Insulin Resistance Zohreh Shoar, Michael J Goldenthal, Francesco De Luca, Elizabeth A Suarez St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA; St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA BACKGROUND[br]In adults, low levels of mitochondrial DNA (mtDNA) and mitochondrial enzyme activity have been associated with obesity; in addition, altered mitochondrial function is thought to play a causal role in the metabolic syndrome, as a defective oxidative phosphorylation may be involved in visceral fat gain and development of insulin resistance. However, there is little evidence on the correlation between mtDNA content, mitochondrial function, and adiposity in children.[br]OBJECTIVES[br]Our goal was to compare mtDNA content and mitochondrial enzyme activity between obese and non-obese children and to correlate mtDNA content with BMI.[br]DESIGN/METHODS[br]Obese (BMI[ge]95% for age and sex) and non-obese children, ages 2-18 yrs were recruited. Anthropometric measurements were taken and BMI Z-score and percentile were calculated. Buccal swabs for mitochondrial enzymes (Complex I and Citrate Synthase) were collected and their activity was measured by using a dipstick methodology combined with spectrophotometric analysis. In obese children only, serum levels of fasting glucose and insulin were measured, HOMA-IR was calculated and nuclear/mtDNA from mononuclear cells was isolated and quantitated by real-time PCR.[br]RESULTS[br]54 obese (30 girls, 24 boys, mean age of 12.3 [plusmn] 3.8 yrs) and 25 control (15 girls, 10 boys, mean age of 10.9 [plusmn] 4.6 yrs) children were recruited. In obese children, complex I and Citrate Synthase levels were lower than those of controls (79.5[plusmn]49.6 unit/min/mg protein vs. 92.1[plusmn]43.1and 11.3[plusmn]4.9 nmol/min/mg protein vs.12.6[plusmn]4.3, respectively); however, this difference did not reach statistical significance (p=0.29 and 0.3 respectively). We found a positive correlation between BMI z-score and the ratio of mt/nuclear DNA (Spearman rho=0.43, p=0.035). No correlation was found between mtDNA and HOMA-IR. Lastly, children with the BMI in the highest quartile exhibited a higher mt/nuclear DNA ratio (1.9[plusmn]1.0) compared to those with a BMI in the lowest quartile (0.7[plusmn]0.7), but again the difference was not statistically significant.[br]CONCLUSIONS[br]Our preliminary findings in obese children suggest that mtDNA content in mononuclear cells is positively correlated with degree of adiposity. Although not statistically significant, the difference of the selected mitochondrial enzyme activity between obese and control children suggests a decreased oxidative phosphorylation in the former. Further studies in a larger population sample are needed to confirm these findings.[br][br]Nothing to Disclose: ZS, MJG, FDL, EAS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1153 53 251 SAT-138 PO28-02 Saturday 190 2012


191 ENDO12L_SAT-139 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Increased Circulating Placental Growth Factor (PLGF) Concentrations in Children and Adolescents with the Metabolic Syndrome Panagiota Pervanidou, Athanassios Akalestos, Despoina Bastaki, George P Chrousos Athens University Medical School, Athens, Greece Background: Childhood obesity is associated with an increased risk for early onset endothelial dysfunction and atherosclerosis. This association is mediated by pathogenetic mechanisms related to the development of the Metabolic Syndrome (MetS) and associated changes in endothelial and inflammation functions. Placental growth factor (PLGF), a member of the vascular endothelial growth factor family, plays an important role in a range of physiologic and pathologic processes,, including stimulating angiogenesis and atherogenic migration of monocytes/macrophages into the arterial wall, and contributions to preeclampsia, cardiovascular disease and tumor progression,. The aim of this study was to investigate differences in PLGF concentrations between children with obesity/metabolic syndrome and age-matched normal weight controls.[br]Materials and Methods: Forty-four (34 males) obese and 33 lean (normal body mass index, BMI) children were assessed at the Childhood Obesity Clinic of our department. Mean age was 11.65 [plusmn]2.01 years. Obesity and metabolic syndrome were defined using the IOTF and IDF criteria, respectively. PLGF was measured using electrochemiluminescence.[br]Results: Obese children (N=44) had similar PLGF concentrations [mean value 11.82 [plusmn]2.97 pg/ml] to those of children with normal BMI [mean value 11.87[plusmn] 3.09pg/ml], while no significant correlations were found between PLGF and BMI z-score. Among the obese children, those with the full MetS (8 children, 7 males) had significantly higher (p=0.014) PLGF [14.47[plusmn]3.47 pg/ml] concentrations than those without the MetS (11.32 [plusmn] 2.63 pg/ml).[br]Conclusions: Although no apparent differences were found in PLGF concentrations between obese and lean children, obese children with the MetS had higher concentrations of PLGF than obese children without the MetS. These findings suggest PLGF may participate in the process of atherogenesis in obese children and adolescents with the MetS.[br][br]Nothing to Disclose: PP, AA, DB, GPC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2016 53 252 SAT-139 PO28-02 Saturday 191 2012


192 ENDO12L_SAT-140 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Using Structural Equation Modeling To Assess the Associations between Obesity and Pancreatic Beta-Cell Function in Adolescents Alice PS Kong, Kai Chow Choi, Ronald CW Ma, Juliana CN Chan The Chinese University of Hong Kong, Hong Kong, Hong Kong; The Chinese University of Hong Kong, Hong Kong, Hong Kong [bold]Aim[/bold][br]Obesity, insulin resistance and lipotoxicity may contribute to the decline in pancreatic beta-cell function (BCF), which in turn leads to glucose intolerance. Here, we aim to explore the complex inter-relationships between these factors in adolescents using structural equation modeling (SEM).[br][bold]Methods[/bold][br]We identified 225 Chinese adolescents aged 12-19 year-old from a territory-wide, population recruited cohort surveyed in Hong Kong. Anthropometric measurements, fasting blood sampling for lipid profile, insulin (to estimate insulin resistant state by calculating homeostasis model assessment, HOMA-IR) and oral glucose tolerance test (OGTT) were performed. The SEM analyses were performed using LISREL 8.8 (Scientific Software International, Inc.) and the parameters were estimated by maximum likelihood method. The following fit indices were chosen to assess the goodness-of-fit of the path: (1) Chi-square statistic to degree of freedom ratio (c[sup]2[/sup]/[italic]df[/italic]), (2) standardized root mean square residual (SRMR), (3) root mean square error of approximation (RMSEA), (4) adjusted goodness-of-fit index (AGFI), (5) comparative fit index (CFI) and (6) non-normed fit index (NNFI).[br][bold]Results[/bold][br]Mean age of the study cohort was 15.0(+/-1.9) years. 45.8% were boys. 65.3% had normal glucose tolerance. The c2 of the final model is 53.5 with df=25, p[lt]0.001. The goodness-of-fit indices (c2/df =2.1, RMSEA = 0.07, SRMR=0.04, AGFI = 0.94, CFI = 0.98, and NNFI=0.96) indicate that the final model is a reasonably good fit to the data. All the path coefficients are significant (p [lt] 0.05). The model explains for 28%, 29%, 44% and 8% of the variance of lipid profile, HOMA-IR, pancreatic beta cell function and two-hour plasma glucose from OGTT respectively.[br][bold]Conclusion[/bold][br]Obesity leads to decline in BCF through interactions with lipids and insulin resistance in adolescents, culminating to the development of glucose intolerance.[br][br]Sources of Research Support: This study was supported by funding from the General Research Fund from the Research Grants Council (CUHK 4055/01M and 467410), Hong Kong.[br][br]Nothing to Disclose: APSK, KCC, RCWM, JCNC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1193 53 253 SAT-140 PO28-02 Saturday 192 2012


193 ENDO12L_SAT-141 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Effect of Body Fat Mass on Fasting and Glucose-Stimulated Insulin Secretory Dynamics in Healthy Men Ali Iranmanesh, Donna M Lawson, Johannes Veldhuis Salem VA Medical Center, Salem, VA; Mayo Clinic, Rochester, MN Background: Insulin secretion in fasting and fed states is episodic, with circulating concentrations mostly contributed by the pulsatile events.[br]Objective: To assess the effect of total body fat and body fat distribution on circulating insulin levels and insulin secretory parameters in normal men during short-term fasting and after oral glucose intake.[br]Subjects: 57 men ages 19-78 yrs and BMIs 20-39Kg/m[sup]2[/sup][br]Study design: Each man studied on 2 separate occasions, and after an overnight fast.[br]Intervention: Ingestion of either 75 grams of dextrose solution or equal volume of water. Sessions were 6.5-hr long, starting at of 0800-0900 hr. Blood was collected at 10-min intervals for glucose and insulin measurements. DXA and single-slice CT of abdomen were used for the assessment of body composition and abdominal fat area.[br]Results: Mean 6.5 hr serum glucose (g) and insulin (i) concentrations were positively correlated (R/P) with % body fat both on water (g: 0.42/0.001; i: 0.42/0.001), and dextrose (g: 0.49/0.0001; i: 0.43/0.0008) days. Deconvolution analysis of insulin time series identified combined contributions of basal (b) and pulsatile (p) events to total (t) insulin secretion, with positive effects of % body fat in fasting (b: 0.42/0.001, p: 0.27/0.04, t: 0.43/0.0009) and fed (0.35/0.008, 0.42/0.001, 0.43/0.0007) states. While not a determining factor in fasted state, body fat had a positive effect on frequency of insulin secretory bursts after dextrose intake. Backward stepwise multiple regression statistics with % body fat, visceral abdominal fat (VAF), and subcutaneous abdominal fat (SAF) as independent variables [underline]identified SAF as a stronger predictor (R/P)[/underline] of circulating insulin concentrations, and corresponding basal and total insulin release both on water (0.46/0.0003, 0.44/0.0006, 0.46/0.0003) and oral dextrose (0.46/0.003, 0.40/0.002, 0.47/0.0003) days. Pulsatile insulin secretion maintained a higher correlation with SAF during fasting (0.38/0.003), but with % body fat (0.42/0.001) after dextrose intake.[br]Summary: Excess body fat in healthy men augments pulsatile and basal insulin release both during short-term fasting and after glucose ingestion, with resulting increases in circulating insulin concentrations. The latter is further augmented by increased number of insulin secretory bursts, but only in the fed state. By virtue of association, SAF rather than VAF appears to be a stronger factor in fat-induced alterations in insulin/glucose homeostasis in men.[br][br]Nothing to Disclose: AI, DML, JV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1722 53 254 SAT-141 PO28-02 Saturday 193 2012


194 ENDO12L_SAT-142 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) Endurance Training Increases Serum Uncarboxylated Osteocalcin Level and Decreases HOMA-IR Via Reduction of Serum Leptin in Obese Young Patients Eun Jin Kwon, Yu Sik Kim, Shinae Kang, Dong Wook Yeo, Min Kyung Kim, Jiyoon Ha, Youngmi Lee, Jiwoon Kim, Tae Woong No, Jong Suk Park, Kyung Rae Kim, Sang Hoon Suh, Chul Woo Ahn Yonsei University Colleage of Medicine, Seoul, Republic of Korea Recently, serum osteocalcin (OC) secreted from osteoblast has been reported to enhance glycemic control. On the other hand, moderate increase of leptin has been known to stimulate osteoblastic functions. Therefore, it can be assumed that leptin may regulate glucose homeostasis by stimulating the secretion of OC. Between two forms of OC (uncarboxylated OC (unOC) and carboxylated OC (cOC)), unOC is the predominant form in the circulation. Interestingly, leptin is known to increase the expression of osteotesticular protein tyrosine phosphatase, which in turn is known to decrease OC secretion by inducing OC carboxylation. This suggests that reduction of leptin may result in increased osteoblastic activity, OC production/secretion, and consequently, glycemic control. This study investigated whether endurance training may regulate circulating levels of ucOC and parameters of glycemic control via leptin reduction. 39 class I obese, normoglycemic young Korean males were randomly assigned to control (C, n=10) or exercise group (Ex, n=29). The subjects in Ex group underwent 8 week-supervised-endurance exercise training (2400Kcal/week). Weight, BMI, waist circumference, % body fat, leptin, fasting plasma insulin, and HOMA-IR decreased significantly. Whereas ucOC and total OC increased in Ex, without significant change of cOC. There was also negative correlation between the change in leptin and ucOC. Total adiponectin decreased but the ratio of high molecular weight adiponectin to total adiponectin increased by 6.6 fold. Fasting plasma glucose and AUC from oral glucose tolerance test showed no significant change. Bone alkaline phosphatase and bone mineral density were unaffected by the exercise training. These results suggest that endurance training may contribute to increased circulating OC and decreased HOMA-IR without any significant change in bone metabolism. Considering that this study was limited to the normoglycemic obese individuals, further study in the prediabetic and diabetic individuals may be warranted.[br][br]Nothing to Disclose: EJK, YSK, SK, DWY, MKK, JH, YL, JK, TWN, JSP, KRK, SHS, CWA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1350 53 255 SAT-142 PO28-02 Saturday 194 2012


195 ENDO12L_SAT-143 POSTER SESSION: Obesity Topics II (1:30 PM-3:30 PM) The Association between 25(OH)D and Adiposity Indices among Preadolescent Children (Aged 7[ndash]9 Years). Hye Ah Lee, Young Ju Kim, Eun Hee Ha, Hwayoung Lee, Hye Sun Gwak, Eun Ae Park, Sujin Cho, Se Young Oh, Eun Hee Ha, Hyesook Park, Hae Soon Kim School of Medicine, Ewha Womans University, Seoul, Republic of Korea; School of Medicine, Ewha Womans University, Seoul, Republic of Korea; School of Medicine, Ewha Womans University, Seoul, Republic of Korea; School of Medicine, Ewha Womans University, Seoul, Republic of Korea; School of Medicine, Ewha Womans University, Seoul, Republic of Korea; Kyung Hee University, Seoul, Republic of Korea [bold]Background and aims:[/bold] The lower 25-Hydroxyvitamin D(25(OH)D) in obese child is being considered as a important public health problem, but there is still under debate for its relationship. Therefore, we aimed to investigate the association between 25(OH)D and adiposity indices [body mass index(BMI), body fat mass(BFM), percent body fat(PBF), waist circumference(WC), upper arm circumference, and skin-fold thickness] among preadolescent child (aged 7-9 years).[br][bold]Methods: [/bold]We followed up subjects at 2011 aged 7 to 9 child, who were part of Ewha Birth [amp] Growth Cohort study, which is a prospective cohort established 2001-2006. The 25(OH)D concentration in blood were measured using Radio immunoassay. We assessed the magnitude of association with vitamin D level and adiposity indices using multivariate regression analysis adjusted for sex, age, birth weight and calories.[br][bold]Results: [/bold]38(17.2%) of the total 221 child were deficiency ([lt]20ng/mL) for vitamin D level, and its prevalence was more higher in boys (25.2%) than girls (8.1%). 25(OH)D level was negatively associated with BMI([italic][beta][/italic]=-0.08, [italic]p[/italic]=0.03), BFM([italic][beta][/italic]=-0.10, [italic]p[/italic]=0.04), and WC([italic][beta][/italic]=-0.24, [italic]p[/italic]=0.02) in any given for sex, age, birth weight and calories, but was not associated with PBF, upper arm circumference. Regarding the risk of overweight ([ge]BMI 85%tile), those who were in lower 25(OH)D group had significantly higher risk of overweight than sufficient group ([italic]p[/italic][sub]trend[/sub]=0.01).[br][bold]Conclusions: [/bold]The lower 25(OH)D level is associated with risk of adiposity indices in general preadolescent child. This study may contribute to add the evidence of an association between Vitamin D level and adiposity in general population.[br][br]Sources of Research Support: Acknowledgements: This work was supported by National Research Foundation of Korea Grant funded by the Korean Government (2010-0026225).[br][br]Nothing to Disclose: HAL, YJK, EHH, HL, HSG, EAP, SC, SYO, EHH, HP, HSK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1382 53 256 SAT-143 PO28-02 Saturday 195 2012


196 ENDO12L_SAT-154 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) GAD65 and IA-2 Autoantibodies Identify a Population of Type 2 Diabetes Patients with a Significant Impairment in [beta]-Cell Function Angela R Subauste, Anne Chang, Cindy Plunkett, Suman Tandra, Massimo Pietropaolo University of Michigan, Ann Arbor, MI Type 2 diabetes mellitus (T2DM) represents a group of heterogeneous metabolic diseases, ranging from abnormalities related to glucose-stimulated insulin secretion, insulin resistance and autoimmune abnormalities of pancreatic [beta]-cells. The objective of this study was to evaluate indexes of insulin secretion and visceral adiposity in clinically diagnosed T2DM patients with and without the presence of GAD65 and IA-2 autoantibodies (Ab). A total of 18 subjects with T2DM diagnosed over the age of 40 participated in this study: 10 GAD65 and IA-2 Ab negative and 8 GAD65 and/or IA-2 Ab positive. These patients were subdivided in two groups: Ab positive: insulin and/or metformin and Ab negative: insulin and/or metformin. None of these individuals had family history significant for T1DM.These subjects underwent an arginine challenge test to asses [beta]-cell function and a DEXA scan to asses body composition. Both groups were matched for age and sex. No statistically significant differences were found between Ab positive vs Ab negative in terms of age (years) (54[plusmn]3 vs 57[plusmn]3), duration of T2DM (years) (11.6[plusmn]2.2 vs 14.7[plusmn]2.0), BMI (33[plusmn]3 vs 30[plusmn]2) or HgbA1c (8.2[plusmn]0.6 vs 7,5[plusmn]0.5). Both groups had similar body composition with DEXA central fat (g) estimated as 3055 [plusmn]299 for GAD65/IA-2 positive and as 3045[plusmn]671 for GAD65/IA-2 negative. DEXA free fat mass (kg) was estimated at 54[plusmn]5 and 54[plusmn]4 respectively. Ab positive T2DM patients exhibited severely impaired [beta]-cell function as demonstrated by lower AIRmax (33 vs. 197 p=0.0431) compared to the Ab negative. In conclusion, GAD65 and IA-2 Ab identify a pathogenetically distinct subpopulation of T2DM patients with similar body mass composition and seemingly immune-mediated injury of pancreatic [beta]-cells.[br][br]Sources of Research Support: NIH grant DK056200 awarded to MP.[br][br]Nothing to Disclose: ARS, AC, CP, ST, MP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2150 54 257 SAT-154 PO11-02 Saturday 196 2012


197 ENDO12L_SAT-155 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Steps toward Adopting Universal Gestational Diabetes Mellitus Criteria: A Review of GDM Practice at the Centre Hospitalier de l[apos]Universite de Montreal (CHUM) Thi Hoang Lan Nguyen, Elisabeth Codsi, Ariane Godbout Centre de Recherche du Centre Hospitalier de l[apos]Universit[eacute] de Montr[eacute]al (CRCHUM), Montreal, Canada; Centre Hospitalier de l[apos]Universit[eacute] de Montr[eacute]al (CHUM), Montreal, Canada Background: New recommendations for gestational diabetes mellitus (GDM) have been recently proposed (1). Review of local rates of adverse pregnancy outcome was warranted before revision of GDM diagnostic criteria in our center. Objective: To establish local demographic profile, review current care provided and analyze the impact of new diagnostic criteria on our population. Methods: An observational retrospective study was carried out among pregnant women with GDM diagnosed at the CHUM between 2005 and 2009. GDM was diagnosed if at least one of the two values on 75g oral glucose tolerance test was above local criteria: fasting plasma glucose [ge]5.0 or 2-hr post [ge]7.8mmol/L. Rates of adverse pregnancy outcomes were established for macrosomia, defined as birth weight (BW) [ge]4kg, [ge]4,5kg and as large for gestational age (BW [gt]90th percentile for gestational age); for intra-uterine growth restriction (IUGR), defined as low BW ([lt]2,5 kg) and as small for gestational age (BW [lt]10th percentile for gestational age); and for caesarean C-section (CS). Results: A total of 1354 pregnant women were included, representing 1407 pregnancies and 1437 newborns. Women mean age and BMI were 31.9yr and 26.6 kg/m2. Overall, 44.2% were Caucasians and reported family history of diabetes, past GDM or previous macrosomia represented respectively 36.5, 19.1 and 10.0% of subjects. All patients were followed by a nutritionist and 62.9% necessitated insulin therapy. For newborns, mean term and BW were 39wks and 3.4kg. Rates of macrosomia defined as BW[ge]4kg, [ge]4.5kg and LGA were respectively 9.3, 1.3 and 9.9%. Rates of low BW and SGA were 6.0 and 7.3%. Primary CS was needed for 17.7% of deliveries and assisted vaginal birth for 10.3%. Shoulder dystocia complicated 1.7% of birth. Overall, GDM prevalence was 15,3% in our cohort. Application of new criteria would decrease prevalence below 12% but the 308 patients that would have been undiagnosed with GDM did not present better pregnancy outcomes than the rest of the cohort. Conclusion: Despite the high-risk population treated at the CHUM, this study demonstrates lower complications rates in our cohort then in others reported GDM populations (2,3) and results were comparable to general population (4-8) suggesting benefice of even lower diagnostic criteria. Universal standardization of diagnostic criteria could improve GDM care but new recommendations have to be adapted to our high-risk population before validation. Further analyses are needed.[br][br](1) International Association of Diabetes and Pregnancy Study Groups Consensus Panel. International Association of Diabetes and Pregnancy Study Groups Recommendations on the Diagnosis and Classification of Hyperglycemia in Pregnancy. Diabetes Care 2010; 33:676-682. (2) Johns K, Olynik C, Mase R, Kreisman S, Tildesley H. Gestational Diabetes Mellitus Outcome in 394 patients. JOGC February 2006, 122-127. (3) Crowther CA, Hiller JE, Moss JR, et al. Effect of treatment of gestational diabetes on pregnancy outcomes. N Engl J Med 2005; 352:2477-86. (4) Statistics Canada. Birth-related indicators (low and high birth weight, small and large for gestational age, pre-term births), by sex, three-year average 2005-2007 in Canada and Quebec. (5) Canadian Institute for Health Information. Too Early, Too Small: A Profile of small babies Across Canada (2009) p. 1-101. (6) Canadian Institute for Health Information. Giving Birth in Canada: Regional Trends From 2001-2002 to 2005-2006, p. 1-48. (7) Canadian Institute for Health Information. Highlights of 2008-2009 Selected Indicators Describing the Birthing Process in Canada, p. 1-7. (8) The Society of Obstetricians and Gynaecologists of Canada (SOGC) website. Quarisma (Quality of Care, Management of Obstetrical Risks and Birthing Mode in Quebec). www.sogc.org/projects/quarisma_e.asp.[br][br]Nothing to Disclose: THLN, EC, AG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 269 54 258 SAT-155 PO11-02 Saturday 197 2012


198 ENDO12L_SAT-156 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Racial Differences in Maternal Metabolic Profiles in Gestational Diabetic (GDM) Women at One Year Postpartum Karen E Elkind-Hirsch, Martha S Paterson, Donna L Shaler, Brett L Schelin, Beverly W Ogden Woman[apos]s Hospital, Baton Rouge, LA; Woman[apos]s Hospital, Baton Rouge, LA [bold]Objective[/bold]: Women who develop gestational diabetes mellitus (GDM) have an increased risk for later glucose intolerance and type 2 diabetes (DM2). We examined whether race impacts the incidence of metabolic disturbances 1 year postpartum in previous GDM (pGDM) women.[br][bold]Design[/bold]: From April 2009-October 2011, 170 pGDM (130 white [W] and 40 non-white [NW]) women underwent metabolic assessment at 2-3 months postpartum including evaluation of glucose tolerance by a 75-g oral glucose tolerance test (OGTT) with glucose and insulin concentrations measured at 0, 30, 60, and 120 min. Participants returned at 12-months postpartum for reassessment. Fasting and mean blood glucose (FBG, MBG), cholesterol (C), HDL- and LDL-C, triglycerides (TRG), liver enzymes, TSH, blood pressure (BP), body mass index (BMI), and waist circumference (WC) were measured. Insulin sensitivity and secretion indices were derived from fasting (HOMA-IR and HOMA-B) and glucose-stimulated levels (Matsuda index [SI[sub]OGTT[/sub]] and [Delta]I[sub]0[ndash]30[/sub]/[Delta]G[sub]0[ndash]30[/sub] [IGI])[sup]1-4[/sup]. A measure of [beta]-cell response to glucose challenge was calculated using the insulinogenic index divided by HOMA-IR to adjust for degree of insulin resistance (IGI/IR).[sup]5,6[/sup][br][bold]Results[/bold]: A total of 113 pGDM women (92 W; 21 NW) returned for follow-up; 21 women on medical therapy for impaired glucose regulation (IGR) diagnosed early (2-3 months) postpartum were excluded from the analyses. Fifty-seven pGDM women were not tested at 12 months (7 DM2; 11 pregnant). The study cohort comprised 92 (73 W; 19 NW) pGDM women studied early and 1 year postpartum. At follow-up, dysglycemia was more prevalent in NW women with 47.4% having IGR compared to 24.7% of W women (p[lt].044). Postpartum weight and WC gain, BP, MBG, and IR were significantly greater whereas [beta]-cell secretory response was significantly decreased in NW pGDM women. W pGDM women had a less favorable lipid profile (higher TRG and LDL-C; p[lt].04). W pGDM women were more likely have breastfed compared with their NW pGDM counterparts (p[lt]02). Patient age, treatment of GDM, parity, FBG, TSH, and liver enzymes were not racially disparate.[br][bold]Conclusions:[/bold] Between 3 and 12 months, pGDM women who progressed to IGR manifest low insulin sensitivity, insulin response and [beta]-cell responsivity. Postpartum weight gain, particularly increased central adiposity, was strongly associated with deterioration of [beta]-cell compensation for insulin resistance which may be a key factor in the greater prevalence of glycemic impairment in NW pGDM women.[br][br](1) Matthews DR et al., Diabetologia. 1985; 28: 412 [ndash]419. (2) Matsuda M et al., Diabetes Care 1999;22: 1462[ndash]1470. (3) Sluiter WJ et al., Diabetes 1976; 25:241-244. (4) Wareham, NJ et al., Diabet. Med., 1995; 12, 931. (5) Kosaka K et al., Diabet Med. 1996; 13(9): S109-19. (6) Hanson RL et al., Am J Epidemiol 151:190[ndash]198.[br][br]Sources of Research Support: Irene W. and C.B. Pennington Foundation and Blue Cross/Blue Shield of Louisiana Impact Grant. (ClinicalTrials.gov number, NCT00849849 [ClinicalTrials.gov]).[br][br]Nothing to Disclose: KEE-H, MSP, DLS, BLS, BWO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 14 54 259 SAT-156 PO11-02 Saturday 198 2012


199 ENDO12L_SAT-157 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Glycemic Control in Familial Versus Sporadic Type 1 Diabetes Patients over 5 Years Sujana Reddy, Steven Reinert, Wendy Plante, Geetha Gopalakrishnan, Charlotte M Boney, JB Quintos The Warren Alpert Medical School of Brown University, Rhose Island Hospital, Providence, RI Background: Previous studies showed that familial type 1diabetes patients (FTID; patients with an affected parent and/or sibling) have less severe metabolic derangement at presentation compared to sporadic patients (ST1D; without affected parent or sibling). Glycemic control was similar at 1 yr follow-up but data on long term metabolic control is lacking.[br]Objective/Hypothesis: 1) FT1D compared to ST1D have less severe presentation at diagnosis and better HbA1c (A1c) over 5 yrs due to parent[apos]s experience in diabetes management 2) In FT1D parent-offspring group, A1c in the child will correlate with parent and 3) in sib-pair group, A1c of the second affected sibling (SP2) will correlate with the first affected sibling (SP1).[br]Methods[bold]:[/bold] Data was collected from T1D children age 6 months to 18 yrs over 20 yr period. Cohort includes 33 parent-offspring and 19 sib-pairs; 33 sporadic patients matched by age, sex, ethnicity, pubertal status, and insulin regimen were selected as controls. We collected A1c, pH, HCO3, glucose at presentation, and A1c, ER and clinic visits and DKA episodes 5 yrs post-diagnosis. Paired T-test and Pearson correlation were used for statistical analysis.[br]Results[bold]: [/bold]At diagnosis, mean age was FT1D (7.7+/- 4.9 yrs) vs ST1D (7.6 +/-4.5 yrs). The metabolic parameters were not different for FT1D vs ST1D (mean A1C: 9.6 vs 10.7%, HCO3: 21 vs 18 meq/l, glucose 428 vs 463 mg/dl and pH 7.35 vs 7.36 (p= ns). In the 5 yr follow up, mean A1C for FT1D vs ST1D (8.9 vs 8.8%; p=0.81), mean clinic (12 vs 12.5, p=0.68) and ER visits (0.48 vs 0.24, p=0.10).[br]Mean glucose at presentation between sib-pair groups (SP1: 433 vs SP2: 347 mg/dl, p=0.49 and HCO3 (SP1:18 vs SP2: 24 meq/l (p[lt]0.01).[br]Mean A1C for SP1 and SP2 was 9.1%. A1C for SP2 correlated positively with that of SP1 (r=0.67, p[lt]0.01). Mean A1C for offspring vs parents (9.3% vs. 8.6 %, p=0.34). A1C for offsprings correlated positively with parents but was not significant (r= 0.57, p=0.18).[br][bold]Conclusion: [/bold]Metabolic control at diagnosis of T1D and the mean A1C over 5 yrs were similar in familial and sporadic patients. In sib-pairs, the second affected sibling had milder clinical presentation compared to the first affected sibling. However, A1C over 5 yrs of SP2 correlates with the SP1. We found only a trend in the correlation of glycemic control between parents and offsprings.Therefore, based on our small sample size, parental experience in diabetes does not positively influence control in their children.[br][br]Nothing to Disclose: SR, SR, WP, GG, CMB, JBQ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2109 54 260 SAT-157 PO11-02 Saturday 199 2012


200 ENDO12L_SAT-158 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) The Effect of Parental Type 1 Diabetes on Glycemic Control in Children with Type 1 Diabetes Alexandra Martin, Michael B Johnson, Whitney Brown, Marielisa Rincon TC Thompson Children[apos]s Hospital, Chattanooga, TN; The University of Tennessee at Chattanooga, Chattanooga, TN; Palmetto Health Children[apos]s Hospital, Columbia, SC [bold]Background:[/bold] Complex psychosocial factors affect control of type 1 diabetes (T1D) in children. Parental T1D has not been identified as an independent predictor of glycemic control. To date, no published data exist comparing control of T1D in children with and without diabetic parents.[br][bold]Objective:[/bold] Demonstrate whether parental T1D is an independent predictor of poor glycemic control in children withT1D, defined as higher hemoglobin A1c (HbA1c), increased incidence of emergency department (ED) visits, increased incidence of diabetic ketoacidosis (DKA), decreased follow-up, and/or decreased frequency of home blood glucose (BG) testing.[br][bold]Design/Methods:[/bold] Data was collected retrospectively over a 3 year period (November 2007-October 2010) from the charts of preadolescent patients with duration of T1D [gt]12 months who were living full time with a parent with T1D. Patients with parental type 2 diabetes were excluded, as were patients age [gt]12 in order to avoid the confounding effects of adolescent behavior. Controls matched for age and sex were randomly selected from patients with T1D who did not have parents or sibling with T1D. Data collected included average HbA1c, ED visits/year, DKA episodes/year, follow-up visits/year, and BG tests/day.[br][bold]Results:[/bold] 15 subjects and 30 matched controls met criteria for inclusion in this study. Patients with parental T1D had trends toward higher mean HbA1c (0.35% higher, p=0.37), greater incidence of DKA (additional 1.44 episodes/year, p=0.16), greater frequency of ED visits (additional 1.99 visits/year, p=0.05), less follow-up (1.64 fewer clinic/visits/year, p=0.11), and decreased home BG testing (1.76 fewer tests/day, p=0.09) as compared to controls. ANCOVA analysis showed that these results were independent of socioeconomic status or duration of T1D.[br][bold]Conclusions:[/bold] All variables showed trends in the hypothesized direction, with parental T1D being associated with a trend toward higher HbA1c, greater incidence of DKA and ED visits, and poorer compliance with follow-up and BG testing. Larger studies are needed to demonstrate statistical significance, and thereafter, to clarify why diabetic children of diabetic parents have poorer control.[br][br]Nothing to Disclose: AM, MBJ, WB, MR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 116 54 261 SAT-158 PO11-02 Saturday 200 2012


201 ENDO12L_SAT-159 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Accuracy of Continuous Glucose Monitoring in Pediatric Patients with Diabetic Ketoacidosis Niyati Skaria, Thomas Wilson, Katherine Messina, Andrew Lane Stony Brook University Hospital, Stony Brook, NY Background[br]Continuous glucose monitors (CGM) are increasingly used in the outpatient setting. Patients may make clinical decisions based on CGM readings, including during development of diabetic ketoacidosis (DKA). Volume contraction during DKA contributes to acidosis via lactic acid production. Because CGM devices rely on measurement of generated hydrogen ions, it is unknown whether DKA affects CGM accuracy.[br]Objective[br]We aim to determine whether a commercially available CGM (Dexcom) measures blood glucose measurements accurately compared to a standard of care bedside glucometer in the setting of diabetic ketoacidosis.[br]Design/Methods[br]Succesive eligible patients ages 2 to 21 years old admitted to our hospital in DKA (venous pH [lt] 7.25) were invited to participate. Venous blood gases and chemistry panels were drawn as per standard PICU protocol. Exclusion criteria: other known metabolic conditions, or dermatologic conditions preventing sensor placement. The CGM was placed subcutaneously at presentation. After warm-up, CGM was calibrated to the bedside glucometer initally, and every 12 hours. CGM values were analyzed by comparison to bedside and serum gluocose values.[br]Results[br]In this ongoing study, 91 paired reference vs. CGM values have been obtained to date from 5 patients, ages 12 to 15 years old. The mean absolute relative difference (MARD) between glucometer and CGM values is 29% (SD=25.7). The MARD between serum glucose and CGM values is 26.7% (SD=24). The MARD between mild, moderate, and severe acidosis groups was not statistically different (p[gt].05). In a Clark Error Grid Analysis- 45% of values fell in category A - desirable correlation and 44% in category B - acceptable correlation. A total of 89% fell in category A and B. Of the rest of the values, 9% fell in category C - risk for inappropriate treatment, 2% in categories D and E - high risk.[br]Conclusion[br]Other investigators studying correlations between CGM and reference values in healthy individuals with diabetes have demonstrated MARDs of 13.2 to 21.5%, and 98% of values falling within Clarke Error Grid category A and B. In contrast, our data suggest that during DKA, there is a much higher MARD between CGM and reference values, and more values fall outside Clarke categories A and B. Our results have important implications in counseling patients and caretakers regarding reliance on CGM values at times of risk for the development of diabetic ketoacidosis.[br][br]Nothing to Disclose: NS, TW, KM, AL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 558 54 262 SAT-159 PO11-02 Saturday 201 2012


202 ENDO12L_SAT-160 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) The Incidence of Vitamin D Deficiency in Children with Type 1 Diabetes Mellitus (T1DM) in Northeast Ohio Berrin Ergun-Longmire, Asuman Bilgin, Lillian Prince Akron Children[apos]s Hospital, Akron, OH [bold]Background:[/bold] Vitamin D deficiency is highly prevalent worldwide although it is largely unrecognized. Several epidemiological studies indicate lower serum 25-hydroxy vitamin D (25OHD) levels in diabetic individuals compared to non-diabetic individuals.[br][bold]Objective:[/bold] The Northeast Ohio is one of the cloudiest places in the world. The aim of our study is to investigate the incidence of vitamin D deficiency in children with T1DM and its correlation with their glycemic control.[br][bold]Methods:[/bold] We conducted a retrospective chart review to identify serum 25OHD levels in patients with T1DM between January 1, 2010 and December 31, 2011. Vitamin D deficiency was defined if serum 25OHD level was [le]20 ng/mL. Vitamin D insufficiency was defined if serum 25OHD level was between 21-31 ng/mL. Levels of HbA1C and vitamin D were stratified in four seasons. The four seasons are defined as follows: Winter, December to February; Spring, March to May; Summer, June to August; and Fall, September to November. We used the SAS 9.2 statistical software for data analyses.[br][bold]Results:[/bold] Data were available from 200 children with T1DM (96 females, 104 males). The mean age, in years, was 12.51[plusmn]3.93. The mean duration of T1DM was 5.12[plusmn]3.31 years. The overall incidence of vitamin D deficiency was 12.5% and insufficiency was 46%. HbA1C was found to be higher in patients with vitamin D deficiency and insufficiency. When analyzing for a correlation using Pearson correlations (r) between HbA1C and serum 25OHD levels, correlations were not significant.[br]A one-way between subjects ANOVA was conducted to compare the effect of the four seasons on serum 25OHD levels. There was a significant effect of the seasons on the Vitamin D25 levels at the p[lt]0.05 level [F(3, 196) = 10.10, p = 0.0001]. The Bonferroni post-hoc comparison test indicated that the mean Vitamin D25 level for the Summer season (34.95[plusmn]9.68) was significantly different than all of the other seasons. These results suggest that highest levels of Vitamin D25 are indeed found in the Summer season. Specifically, the results confirm that the levels decrease during seasons of less sunlight. However, there was no significant difference in the mean Vitamin D25 levels for the pair-wise comparisons between the other seasons.[br][bold]Conclusion: [/bold]Although vitamin D deficiency/insufficiency is common in children with T1DM, there is no statistically significant correlation between their glycemic control and serum vitamin D levels.[br][br]Nothing to Disclose: BE-L, AB, LP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1772 54 263 SAT-160 PO11-02 Saturday 202 2012


203 ENDO12L_SAT-161 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Type 2 Diabetes Mellitus Re-Emergence Post Gastric Bypass Surgery Yessica Ramos, Sophie Bersoux, Lori R Roust, Yu-Hui H Chang Mayo Clinic Arizona, Scottsdale, AZ Roux-en-Y gastric bypass surgery (RYGB) is an effective intervention for improvement and remission of type 2 diabetes mellitus (T2DM). Limited data exist about re-emergence of T2DM after this procedure. A retrospective analysis of patients who underwent RYGB from January 2000 to December 2007 at our institution was performed. 138 patients with documented T2DM were identified. Of these, 72 patients (52%) had minimum 3-year continuous follow-up at our institution. The 72 patients were assessed pre- and post-operatively at various time intervals. Patients[apos] baseline demographics, weight, body mass index (BMI), HbA1C, fasting glucose and diabetic pharmacological treatment were documented. The purpose of the analysis was to determine the persistence, remission and re-emergence of T2DM. Remission of T2DM was defined as HbA1c of less than 6.5% with no pharmacological treatment. Re-emergence was defined by HbA1c of equal to or [gt]6.5%, fasting glucose [gt] 7mmol/L or re-initiation of anti-diabetic medications. The patients[apos] mean pre-operative BMI was 45 kg/m2. The majority were Caucasian (93%), mean age was 49.5 +/- 9.3 years, and 54% were female patients. The mean duration of T2DM preoperatively was 4.9 +/- 4.4 years. 66 patients (91.6%) were in remission at a minimum of one time point while 6 patients (8.4%) had persistence of T2DM throughout follow-up. Of the 66 patients that had remission, 14 patients (21.2%) had recurrence of T2DM. Five of these patients developed T2DM at two years post RYGB, 3 at 3 years, 3 at 4 years and 3 at 5 years. Duration of T2DM prior to RYGB was significantly associated with recurrence of T2DM (p=0.0021). The odds ratio was 1.26 indicating that longer duration of pre-op T2DM was associated with higher probability of recurrence. No significant association between pre-op BMI or weight regain was identified with a higher recurrence rate. Conclusion: The recurrence rate of T2DM after RYGB is 21%. Duration of diabetes prior to RYGB was associated with a higher recurrence rate of T2DM. This suggests that early surgical intervention in the type 2 diabetic obese population may improve durability of remission of T2DM.[br][br]Nothing to Disclose: YR, SB, LRR, Y-HHC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1881 54 264 SAT-161 PO11-02 Saturday 203 2012


204 ENDO12L_SAT-162 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) FPG, OGTT, HbA1c: Which Combination Best Detects Diabetes Mellitus? Daryl Wei Chong Tan, Daniel Ek Kwang Chew Tan Tock Seng Hospital, Singapore, Singapore Background: Previous methods of diagnosing diabetes mellitus in Singapore were the fasting plasma glucose (FPG) and the 2-h 75g oral glucose tolerance test (OGTT). However, the American Diabetes Association (ADA) has recommended the use of HbA1c as an alternative test for diabetes mellitus with a threshold of [ge] 6.5% in 2010. These diagnostic tests detects differing pathophysiology of dysglycemia amongst diabetes subjects and as such do not demonstrate complete concordance. This study aims to determine which combination of the three tests best detects diabetes mellitus.[br]Methods: Data of subjects who had HbA1c, FPG and OGTT tests done in the same seating between January 2007 and March 2010 at Tan Tock Seng Hospital Diabetes Centre were collected and analysed. Subjects with FPG values [ge] 7.0mmol/l, OGTT values [ge] 11.1 mmol/l, or HbA1c [ge] 6.5% were classified as diabetes mellitus.The records of 250 subjects were used.[br]Results: Of the 250 subjects in the study, 133 (53.2%) had newly diagnosed diabetes mellitus defined by a high FPG or OGTT or HbA1c. OGTT has the highest detection rate of diabetes mellitus (82.7%), followed by HbA1c (62.4%) and then FPG (52.6%). The combination of tests with the highest detection rate is that of HbA1c plus OGTT (97.7%), higher than the combination of FPG plus OGTT (89.5%) or FPG plus HbA1c (72.9%).[br]Conclusion: The combination of HbA1c and OGTT has a highest detection rate of diabetes mellitus out of the three tests FPG, OGTT and HbA1c.[br][br]Nothing to Disclose: DWCT, DEKC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 295 54 265 SAT-162 PO11-02 Saturday 204 2012


205 ENDO12L_SAT-163 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Vitamin D Deficiency across the Spectrum of Glucose Intolerance in an Adult Population at a Northeast Florida Clinic Arnold Vera, Paul Yosmin Casanova-Romero, Sean Patrick Hassett Vera Endocrine Associates, Inc, Daytona Beach, FL Background: The United States is facing a high prevalence of glucose intolerance and diabetes mellitus (DM). Although obesity is clearly a predominant factor in the pathogenesis of diabetes, other modifiable lifestyle factors and certain nutritional factors, such as vitamin D deficiency, are also believed to play a role. The aim of this study was to investigate[bold] the proportion of individuals with vitamin D 25-(OH) insufficiency/deficiency (VitD-)[/bold] across the spectrum of glucose status in an adult population referred for endocrine evaluation at a Northeast Florida clinic.[br]Methods: We performed a comprehensive history, physical and laboratories on consecutive subjects referred for an endocrine evaluation to determine the prevalence of VitD- and its relationship with age, BMI, glucose status, renal status by GFR categories, serum PTH, calcium, phosphorus and magnesium. A Chi-squared test was used to test the proportions for each group.[br]Results: 1663 consecutive subjects aged 18-98 and a mean BMI of 31.42 were evaluated. 51.9 % were female. 1077 individuals had a glucose abnormality: 28.0 % had impaired fasting glucose (IFG) or impaired glucose tolerance (IGT), 55.8 % were diagnosed with Type 2 DM and 16.2 % with Type 1 DM. Females diagnosed with IFG/IGT were found to have a significantly higher proportion of VitD- (40.7 %) than subjects with diagnosis of Type 1 (25.3 %) or Type 2 DM (29.0 %), independent of their age, gender and BMI (Chi-square 7.33 df 2 p[lt]0.05). Adults [lt] 55 year/old with Type 2 DM have a higher proportion of VitD- (42.6%) than subjects 55 to 75 year/old (33.9 %) or [gt] 75 year/old (21.1 %) (Chi-square 12.44 df 2 p[lt]0.001). Obese individuals diagnosed with IFG/IGT (73.3 %) and Type 2 DM (35.3 %) have the highest proportion of VitD- compared with normal or overweight subjects, independent of GFR categories (Chi-square 11.7 df 1 p[lt]0.001; Chi-square 6.9 df 1 p[lt]0.01, respectively).[br]Conclusions: Our data suggests that there is an increased prevalence of vitamin D insufficiency/deficiency in the Southeast female population of the USA with diagnosis of IFG/IGT. Obese adults [lt] 55 year/old with IFG/IGT experience the highest prevalence of VitD- independent of their renal status. We conclude that there is a need to do an early analysis and adjustment of vitamin D levels in adult obese individuals with diagnosis of pre-diabetes; future studies will determine if vitamin D treatment will delay the onset of diabetes.[br][br]Nothing to Disclose: AV, PYC-R, SPH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1530 54 266 SAT-163 PO11-02 Saturday 205 2012


206 ENDO12L_SAT-164 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Assessment of Pain and Treatment Satisfaction in Patients with Painful Diabetic Peripheral Neuropathy Latha Dulipsingh, Susan Zailskas, Teresa McInnis, Aniello Marotta The Hospital of Central Connecticut, New Britain, CT; Pfizer,Inc, New Haven, CT The purpose of this project was to describe pain experienced from painful diabetic peripheral neuropathy (pDPN) and evaluate current treatment satisfaction in patients followed through an endocrinology clinic.[br]Patients [ge] 18 years of age with a diagnosis of diabetes, with or without diabetic peripheral neuropathy, were offered a voluntary self-administered questionnaire, prior to their examination by the healthcare provider. Only patients who had nerve pain were asked to complete the entire questionnaire. In addition to basic information related to diabetes, patients were asked if they experienced nerve pain, the characteristics of the nerve pain, and questions related to treatment and satisfaction with current therapy.[br]A total of 98 questionnaires were collected with 53.1% of the patients being female in gender. The mean age was 55.1 years, 75.6% had type 2 diabetes and the mean duration of diabetes was 16.3 years. Thirty-one patients (31.6%) reported neuropathic pain with 83.3% having experienced nerve pain for one year or greater and the majority (67.7%) reporting nerve pain in their feet. The percentage of patients who reported moderate to severe nerve pain, score of [ge]4 on the 0-10 numeric pain rating scale, over the past week was 54.8% with 32.3% reporting nerve pain [ldquo]All Day [ldquo] in the past 24 hours. Impact of nerve pain on quality of life was noted by 61.3% of patients although 64.5% of patients reported that nerve pain did not cause them to avoid activities. Fifteen (48.4%) patients reported receiving medication for their nerve pain of which 66.7% reported being satisfied with their treatment. Of the satisfied patients, 40% reported severe nerve pain over the past week with 30% reporting pain [ldquo]All Day[rdquo].[br]Painful diabetic peripheral neuropathy is a common complication of diabetes mellitus and can have debilitating consequences with a significant impact on quality of life. Based on our questionnaire we found about a third of patients with long standing diabetes having pDPN of which about two thirds of them reported that it affected their quality of life. Surprisingly, almost half of the patients reported being satisfied with their treatment despite severe nerve pain.[br]It is important for healthcare providers to recognize barriers to pain control in patients with pDPN. Our findings will serve as a foundation for a pDPN provider-patient initiative focused at improving education and communication at our clinic.[br][br]Nothing to Disclose: LD, SZ, TM, AM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1569 54 267 SAT-164 PO11-02 Saturday 206 2012


207 ENDO12L_SAT-165 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Comparison of the Adherence to Guidelines of Diabetes Care in Primary Care and Subspecialty Clinics Lily K Sunio, Deepthi T Rao, Ved Gossain Michigan State University, East Lansing, MI Background:[br]Diabetes mellitus, with an estimated US prevalence of 8.3%, is a major public health problem with significant morbidity and mortality. Evidence based guidelines have been proposed to reduce the micro and macrovascular complications, but studies have shown that these goals are not being met.[br]Objective:[br]To compare the adherence to American Diabetes Association guidelines for measurement and control of glycohemoglobin (A1c) blood pressure (BP), lipids(LDL) and microalbuminuria (MA) by subspecialty and primary care clinics in an academic medical center.[br]Methods:[br]390 randomly selected charts of patients with diabetes mellitus from diabetes (DM), Internal Medicine (IM) and Family practice (FP) clinics at Michigan State University were reviewed. We assessed measurement and control of A1C, LDL, BP and MA during the one year period preceding their most recent visit.[br]Results:[br]Our study included 131, 134 and 125 charts from the FP, IM and DM clinics, respectively. Mean age of the subjects was 61[plusmn]11 (FP), 62.11[plusmn]12.88 (IM) and 52.82[plusmn]16.01 (DM) years. DM clinic had a higher percent of type 1 diabetes (34.4%) compared with about 5% in FP and IM clinics. A1C was measured in 99.2, 97.8, 100% of patients and was controlled ([lt]7%) in 38.9, 43.3, 28.8% of patients in the FP, IM and DM clinics, respectively. LDL was measured in 93.9, 91.8, 89.6% of patients and was controlled ([lt]100mg/dL) in 71.8, 64.9, 64% of patients in the FP, IM and DM clinics, respectively. MA was measured in 85.5, 82.8, and 76.8% of patients in the FP, IM and DM clinics, respectively. BP was measured 100% of the time in all clinics. 59.5, 67.2, 52.8 % of patients in the FP, IM and DM clinics, respectively had their BP controlled ([lt]130/80). Insulin use was associated with poor glycemic control. Patients on insulin were more likely to have A1C[ge]7 % (OR=4.37, p=0.00). Patients with uncontrolled BP were more likely to have LDL[ge]100mg/dL (OR=1.8, p=0.02). Patients on diuretics were more likely to have BP[gt]130/80 (OR=1.7, p=0.05).[br]Conclusion:[br]Our data show that the rate of HbA1C, LDL, and BP testing was high, similar to the national average in both subspecialty and primary care clinics. However, the degree of control of these parameters was still suboptimal which could be influenced by several factors. Continued efforts to improve these parameters are still needed.[br][br]Nothing to Disclose: LKS, DTR, VG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2086 54 268 SAT-165 PO11-02 Saturday 207 2012


208 ENDO12L_SAT-166 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Remission in Minorities with Newly Diagnosed Diabetes Mellitus Treated in a Real World Setting Abhishek Kansara, Yin Oo, Pranav Ghody, Anuja Kriplani, Alejandra Borensztein, Rajeev Sharma, Mikhail Kagan, Mary Ann Banerji State University of New York (SUNY) Downstate Medical Center, Brooklyn, NY Objective[br]Remission in type 2 diabetes is described in African-Americans, Hispanics, Asians and Caucasians with a frequency ranging from 20 to 50 % in a research setting. Our goal was to determine the frequency of remission in newly diagnosed minority diabetes patients treated in a real world clinic setting. We examined clinical factors influencing remission.[br]Research Design and Methods[br]We retrospectively analyzed data of 87 newly diagnosed diabetes patients, seen regularly in diabetes or primary care clinic. Therapeutic decisions were individualized. Remission was defined as achieving normoglycemia with A1C [le] 6.3% on medications. We expect that true remission (off all medications) would come from this subset. Remitters and non-remitters were analyzed by chi square and independent samples[apos] t-tests. Data are mean[plusmn]SD.[br]Results[br]39 out of 87 (45%) achieved normoglycemia while 48 (55%) did not. The baseline characteristics of the remission and non-remission group were not statistically different in terms of age (48[plusmn]12 years), gender (2/3 male) or BMI (26.7[plusmn]12). The initial A1C was lower in the remission group (10.6[plusmn]2.9% vs 12.1[plusmn]2.8%; p=0.015). The initial plasma glucose and clinically relevant microalbuminuria were both significantly higher in the remission vs the non-remission group. The follow-up time was 12.8[plusmn]13.0 months.[br]Patients attending diabetes clinic were more likely to develop remission than those in primary care clinic (67% vs 29%, p=0.014). Treatment using insulin or insulin plus oral anti-diabetic agents was twice as effective as those treated with oral agents alone (63.2% vs 30.6%, p=0.004). Clinicians in diabetes clinic were more likely to prescribe insulin based treatment compared to those in primary care clinic (53% vs 12%, p=0.0001). The time to achieve remission was 4.5 [plusmn]2.3 months.[br]Summary[br]Nearly half (45%) of minority patients in a real world setting achieve near normoglycemic remission. Key determinants of such imporved glucose homeostasis are treatment in a diabetes clinic and the use of insulin at the outset which doubled the remissions compared to primary care clinic. Prolonged near-normoglycemia is known to prevent diabetes complications, thus, inducing remissions is likely to have far reaching public health benefits in reducing morbidity. Our data suggest that early referral of new onset diabetes to a specialist and the use of insulin based treatment optimizes the likelihood of inducing remission.[br][br]Disclosures: MAB: Clinical Researcher, Merck & Co.; Scientific Board Member, Novartis Pharmaceuticals; Speaker Bureau Member, Sanofi-Aventis, Merck & Co. Nothing to Disclose: AK, YO, PG, AK, AB, RS, MK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2374 54 269 SAT-166 PO11-02 Saturday 208 2012


209 ENDO12L_SAT-167 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Deterioration of Diabetes Glucose Control during Civil Unrest M Loutfy Sakkal, Saad Sakkal Metabolic Care Center, Aleppo, Syrian Arab Republic; University of Kalamoon, Deir Atieh, Syrian Arab Republic Context: Civil unrest is stressful to all citizens[apos]. Stress effect diabetes control in general, but no or rare studies showed the influence of civil unrest on diabetes glucose control. We quantitated the effect during the recent Syrian civil unrest.[br]Methods:[br]In our clinic patients are seen quarterly. In each visit we review H[amp]P, laboratory parameters and undercurrents psychosocial factors.[br]In 200 usual clinic patients we determined means FBS, 2HPP, HgA1c during 2 distinct comparable periods: for the 6 months before the civil unrest and after (July to December 2010, 2011).We excluded new onset diabetes patients and other illnesses for a separate reports.[br]Results:[br]In the period before civil unrest July to December 2010 There was a drop in FBS: 37 mg/dl (177 to 140), 2hpp 62(214 to 152), HgA1c:2.5 %(%9.9 to 7.4) and insulin dose by 22 %(50 to38) from baseline. In the period after civil unrest July to December2011 there was a rise in FBS: 47(137 to 184), 2hpp:128(161 to 289), HgA1C:2%%7.6 to 9.6) and insulin dose by 25 %(44 to 55).[br]Conclusion:[br]Diabetes Glucose control is affected by civil unrest, probably as a result of stress, lack of health resources or other factors. We need more studies for diabetes during unfortunate civil unrest events.[br][br]Nothing to Disclose: MLS, SS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 155 54 270 SAT-167 PO11-02 Saturday 209 2012


210 ENDO12L_SAT-168 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Response to Vitamin D Supplementation in Type 2 Diabetes Patients with Pulmonary Tuberculosis Sunil K Kota, Siva K Kota, Svs Krishna, Lalit K Meher, Kirtikumar D Modi Medwin Hospital, Hyderabad, India; Central Security Hospital, Riyadh, Saudi Arabia; MKCG Medical College, Berhampur, India Objective:To evaluate effects of vitamin D supplementation on type 2 diabetes mellitus (T2DM) patients with pulmonary tuberculosis(PTB).[br]Methods:45 subjects (M:F= 34: 1) were screened.Inclusion criteria :Age[gt]15 years,newly diagnosed PTB cases with uncontrolled T2DM, serum vitamin D[lt]20 ng/ml.The patients were randomly assigned to 2 groups.Group 1 subjects received oral cholecalceferol(60000 units/week) and calcium carbonate (1 gm/day) along with anti tubercular treatment(ATT),while group 2 subjects did not. Sputum was checked at interval of 2 weeks for 12 weeks.Primary end point was time to achieve sputum smear conversion.Secondary end points were reduction in ESR and improvement in glycemic parameters.[br]Results:15 patients with vitamin D[gt]20 ng/ml were excluded.So the prevalence of vitamin D deficiency in T2DM with PTB was 30/45 (66.66 %),with 25/34 males (73.5%)and 5/11 females (45.5%)were vitamin D deficent.Mean age of patients (30) was 39.5[plusmn]18.9 years with FBS 230.5[plusmn]30.3 mg/dl,PLBS 320.5[plusmn]45.6 mg/dl,HbA1C 10.4[plusmn]4.4 % and 25(OH)D 12.1[plusmn]4.3 ng/ml. At end of 12 weeks,group 1 patients had significantly higher levels of serum 25(OH)D (25.4[plusmn]6.9 ng/ml in group 1 versus 10.2[plusmn]0.9 ng/ml in group 2).Sputum smear conversion was 6 weeks in group 1 versus 8 weeks in group 2(p= 0.067).Reduction in ESR was significant in group 1 vs group 2 (39.6 [plusmn] 12.4 mm/1st hr vs 24.0 [plusmn] 14.9, p-0.004).Difference in the reduction in FBS, PLBS and HbA1C in the 2 groups did not attain statistical significance.[br]Discussion:Correlations exist between low vitamin D levels with PTB and T2DM separately.Calcitriol modulates response to mycobacterial infection by induction of reactive nitrogen and oxygen intermediate,suppression of matrix metalloproteinase enzymes implicated in pulmonary cavitation, and induction of antimicrobial peptide cathelicidin.Calcitriol modulates immune responses by binding vitamin D receptors expressed by antigen presenting cells and activated lymphocytes. Several case series have reported utility of 25000 IU to 100000 IU vitamin D in improving patients[apos] response to ATT (1). Vitamin D supplementation also increases lymphocyte to monocyte ratio,a biomarker of disease resolution.We provided vitamin D at the dosage of 60000 IU/week, and report a 2 weeks reduction in sputum smear conversion.[br]Conclusion: Vitamin D may be the missing link between emerging epidemic of tuberculosis [amp] diabetes.It can serve as adjuvant treatment of tuberculosis in diabetics with vitamin D deficiency.[br][br]1. Martineau AR, et al., J Steroid Biochem Mol Biol 2007; 103: 793.[br][br]Nothing to Disclose: SKK, SKK, SK, LKM, KDM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1403 54 271 SAT-168 PO11-02 Saturday 210 2012


211 ENDO12L_SAT-169 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Impairment in Glycemic Control and Lipid Profile during Indian Festival Diwali Nitish Mathur, Rachita Mathur, Jairam Rawtani Dr S N Medical College, Jodhpur, India; Dr S N Medical College, Jodhpur, India INTRODUCTION: The effect of cultural factors on glycemic control in Asian Indian diabetics has not been studied in detail. Hindu festival of Diwali is associated with increased consumption of sweets and oil rich fried dishes and likely to influence metabolic control. Therefore, present study aims at estimating the changes in glycemic control and lipid profiles after Diwali.[br]METHOD: The study was conducted on 74 T2DM patients (age 60[plusmn]8.26 years, M:F ratio 43:24), recruited from out-patient department of Dr. S. N. Medical College, Jodhpur, India.T2DM was diagnosed as per ADA criteria, and all the subjects were on oral anti-diabetic drugs. Blood samples were collected 2 weeks before and after the Diwali. The patients were instructed to continue with the same oral anti-diabetic medication during this period. Following parameters were measured from the samples: Fasting and post-prandial plasma glucose, Lipid profile (serum total cholesterol, triglycerides, HDL, LDL and VLDL cholesterol)[br]RESULTS: Out of total 74 patients, 67 patients returned for follow-up. There was a significant increase in fasting (136.85[plusmn]2043 vs.154.02[plusmn]19.16, p = 1.82 [times] 10[sup]-14[/sup]), and post-prandial (185.22[plusmn]30.66 vs. 207.05[plusmn]31.16, p = 8.63 [times] 10[sup]-14[/sup]) plasma glucose, total cholesterol (198.47[plusmn]18.84 vs. 212.98[plusmn]19.06, p = 1.69 [times] 10[sup]-15[/sup]), triglycerides (136.37[plusmn]25.76 vs. 153.44[plusmn]23.95, p = 1.18 [times] 10[sup]-15[/sup]), LDL (130.25[plusmn]16.42 vs. 142.28[plusmn]17.50, p = 4.56 [times] 10[sup]-13[/sup]) and VLDL (27.27[plusmn]5.15 vs. 30.68[plusmn]4.79, p = 1.18 [times] 10[sup]-15[/sup]) cholesterol after Diwali. Whereas, HDL was found to be reduced significantly after Diwali (40.94[plusmn]3.75 vs. 40.01[plusmn]2.78, p = 3.57 [times] 10[sup]-4[/sup]).[br]CONCLUSION: Diwali is associated with significant impairment in glycemic control and lipid profile. Therefore, around the festive season, prescription must be adjusted in accordance with the change in food intake and patients must be provided with specialized diet counseling for the season.[br][br]Nothing to Disclose: NM, RM, JR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1181 54 272 SAT-169 PO11-02 Saturday 211 2012


212 ENDO12L_SAT-170 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Glucose Control during a Driving Training in Patients with Diabetes Peter Wiesli, Joel Capraro, Beat Frauchiger, Giatgen A Spinas, Roman Truninger Kantonsspital Frauenfeld, Frauenfeld, Switzerland; University Hospital of Zurich, Zurich, Switzerland [bold]Objective[/bold]: To investigate the effect of prolonged acute mental stress by means of a driving training on glucose control in patients with type 1 and type 2 diabetes.[br][bold]Research Design and Methods[/bold]: 39 patients with insulin-treated diabetes (18 type 1, 21 type 2 diabetes) were exposed to mental stress by means of a 2hr-driving training. The training session started 15 min after intake of a standard meal. Plasma glucose, blood pressure, heart rate, salivary cortisol, and stress perception (by visual analog scale) were monitored in regular intervals and compared to a control day either preceding or following the stress testing day, respectively.[br][bold]Results[/bold]: On the stress testing day, blood pressure rose from 142/86 [plusmn] 16/9mmHg at baseline to 157/93 [plusmn] 17/10mmHg (p[lt]0.05), heart rate from 72 [plusmn] 11bpm to 84 [plusmn] 15bpm (p[lt]0.05), stress perception from 1.4 [plusmn] 0.6 to 4.1 [plusmn] 1.9 points (p[lt]0.05), and salivary cortisol concentrations from 5.8 [plusmn] 3.3nmol/l to 7.6 [plusmn] 3.9nmol/l (p[lt]0.05); these measurements remained stable on the control day. In the entire study group, the course of glucose concentrations showed no significant difference on the stress testing day compared to the control day. However, 67% of patients (26/39) displayed a change of glucose concentrations of more than 2mmol/l on the stress testing day as compared to the control day; with 46% of patients (18/39) showing an increase of [gt]2 mmol/l (median 3.9mmol/l, range 2.0 [ndash] 8.9mmol/l, p[lt]0.05), and 21% of patients (8/39) showing a decrease of [gt]2mmol/l (median 4.5mmol/l, range 2.7 [ndash] 11.0mmol/l, p[lt]0.05), respectively. 33% of patients (13/39) experienced no relevant alteration of blood glucose ([lt]2mmol/l).[br][bold]Conclusions[/bold]: Although a majority of patients with diabetes display clinically relevant deviations of blood glucose control during prolonged mental stress, there is no constant pattern of glucose dysregulation.[br][br]Nothing to Disclose: PW, JC, BF, GAS, RT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 127 54 273 SAT-170 PO11-02 Saturday 212 2012


213 ENDO12L_SAT-171 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Use of Professional Continuous Glucose Monitoring (PCGM) in Outpatients with Diabetes at a Community Hospital Keya Malhotra, Sally M Pinkstaff, Naomi Walpert Sinai Hospital of Baltimore, Baltimore, MD [bold]Background[/bold]: Glucose monitoring is recommended for most patients with diabetes. Two techniques are available: self-monitoring of capillary glucose (SMG), and continuous monitoring of interstitial glucose (CGM). Intermittent use of CGM has been used when glucose control is not achieved. Recent AACE and Endocrine Society guidelines have outlined uses for PCGM, including suspected hypoglycemia (hypo), A1C not at target, postprandial hyperglycemia (hyper), or major medication change. The diabetes center (DC) at Sinai Hospital has used PCGM for 3 years. We conducted this chart review to evaluate the patient population referred for PCGM and the results of the PCGM tracings. Objectives: To assess the validity of the PCGM profiles, to compare the reasons for referral with the guidelines, and to identify the frequency of hypo/hyper excursions.[br]Design: Retrospective chart review.[br]Setting: Outpatient DC.[br][bold]Methods[/bold]: 98 patient profiles from January-June 2010 were downloaded using PCGM software. A valid PCGM profile required: 1)[gt]260 readings on each of 3 consecutive days, 2)[gt]2 paired SMG readings/day, 3) Mean Absolute Difference [lt]28%, and 4) R[gt]0.79. Frequency of hyper ([gt]180 mg/dl) and hypo ([lt]70 mg/dl) was calculated from a consecutive 72 hour tracing. 52 patient records with valid PCGM profiles were reviewed.[br][bold]Results[/bold]: Fifty-two PCGM profiles met the criteria for validity. 12 patients had type 1 diabetes (T1D), 40 had type 2 diabetes (T2D), and 37 had an A1C[gt]7%. 43 patients were on insulin. Testing indications were: elevated A1C, suspected hypo, change in medical regimen, dawn phenomenon, or postprandial hyper. PCGM tracings showed hyper 0-20% of the time in 4 T1D patients and 20 T2D patients. Six T1D patients and 13 T2D patients had hyper 21-50% of the time. Overall, hyper for [gt]20% of the time was detected in 67% of T1D and 50% of T2D. No hypo was detected in 2 of T1D and 25 of T2D patients. Hypo for [gt]10% of the time was found in 50% of T1D and 14% in T2D. 4 and 10 of T1D and T2D patients respectively, had hypo between 1-10% of the time.[br][bold]Conclusions[/bold]: This pilot study shows that use of PCGM in our DC needs revision. 50% of tracings were invalid, suggesting inadequate patient training. Reasons for referral agreed with current guidelines in most patients. PCGM profiles did demonstrate frequent hypo and hyper episodes in many patients and can be used to guide medication changes.[br][br]Nothing to Disclose: KM, SMP, NW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1798 54 274 SAT-171 PO11-02 Saturday 213 2012


214 ENDO12L_SAT-172 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) The Development of a Simplified and Integrated Biosensor for Diabetics Francis A Eusebio, Teagan Adamson, Curtiss B Cook, Jeffrey T La Belle Arizona State University, Tempe, AZ; Mayo Clinic Arizona, Scottsdale, AZ Self-monitoring of blood glucose (SMBG) is standard of care in diabetes management. Current technologies for SMBG are based upon enzymatic electrochemical sensing. To increase the sensitivity and specificity of current devices, we are developing a novel method of detecting glucose using electrochemical impedance spectroscopy (EIS) technology. EIS is a frequency based electrochemical method similar to amperometric methods but with the addition of an alternating current (AC) signal (1). This label free technique is capable of detecting femtoMolar concentrations of the desired marker. To test the ability of EIS methods to detect glucose, the enzyme glucose oxidase (GOx) was fixed to gold electrodes through the means of a specific immobilization process. This process involved a self-assembling monolayer, a 16- mercaptohexadecanoic, an N-hydroxysulfosuccinimide and 1-Ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride coupling intermediate, and a blocking element to prevent the attachment of interferents. Once GOx was fixed to the gold electrode surface, a 5 mV sine wave sweeping frequencies from 100 kHz to 1 Hz was induced at glucose concentrations of 0, 1, 4.5, 8, 10, 25, 60, 85, and 100 mg/dL mixed with a ferricyanide redox mediator. Each frequency in the 100 kHz range sweep was analyzed for highest response and r-squared value. The frequency with both factors optimized is specific for the glucose-GOx binding interaction, and was determined to be 966.8 Hz. Four separate electrodes were constructed and data from each were averaged. The mean lower limit of detection was a glucose concentration of 14.6 mg/dL with a relative standard deviation of 14.7 %. The correlation between the impedance response and concentration was determined to be 11.5*ohm/log(mg/dL) with a r-squared value of 0.8. The above data confirm that EIS offers a new method of glucose detection, and development of a sensor strip utilizing EIS methods offers a potential alternative technology for SMBG that offers improved detection at lower concentrations of glucose. The unique frequency response of individual markers allows for modulation of signals so that several markers could be measured with a single sensor. Future work includes assessment of other diabetes associated biomarkers that can be measured on a single sensor, integration testing and tuning of the biomarkers, impedance-time sensing development, and finally, testing on diabetes (and control) subjects.[br][br](1) La Belle JT, Demirok UK, Patel DR, Cook CB. Development of a Novel Single Sensor Multiplexed Marker Assay. Analyst, 2011 136:1496-1501.[br][br]Disclosures: CBC: Coinvestigator, The Epsilon Group. Nothing to Disclose: FAE, TA, JTLB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 400 54 275 SAT-172 PO11-02 Saturday 214 2012


215 ENDO12L_SAT-173 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Capillary and Venous Blood Samples Analyzed in Glucometers Compared to a Laboratory Hexokinase Method Milena G Teles, Suzimara A Oliveira, Isio Schulz, Nairo M Sumita, Maria Elisabeth Rossi, Marcia Nery Hospital das Cl[iacute]nicas da Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Hospital das Cl[iacute]nicas da Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Blood glucose meters are widely used for monitoring diabetes. A question common to both health professionals and patients is how much glucose meters values actually correlate with laboratory values. [bold]Aim: [/bold]To evaluate the performance of two brands of glucometer using both capillary and venous samples compared to plasmatic glucose (PG) analyzed in a laboratory. [bold]Patients and[/bold] [bold]Methods[/bold]: 237 capillary and venous blood specimens of 188 women and 45 men (age 14-87 years) were analyzed on both Accu-Chek Active (ACA)-Roche Diagnostics and Optium Xceed (OX)-Abbott Diabetes glucometers and compared with plasma glucose analysis in a clinical laboratory (hexokinase method). All blood samples were collected by the same trained meter operator. Venous samples were obtained by venipuncture and collected in tubes with fluoride. The time between capillary and venous samples collection was [lt] 3 minutes. After antisepsis with alcohol, the first drop of capillary blood was discarded and the second and third drops were analyzed in each meter in alternate days. [bold]Results[/bold]: The PG values range was 42-447 mg/dL. The correlation coefficients (CC) between capillary samples analyzed in the two devices and PG were similar: 0.973 (OX) and 0.950 (ACA). The mean percentage variation (MPV) regarding capillary and plasmatic levels was 7.84 and 8.39 for OX and ACA, respectively. Concerning venous samples analyzed in the meters, the CC was 0.931 (OX) and 0.978 (ACA) and MPV was 15.45 and 5.94 for OX and ACA, respectively. There was no significant statistical difference in relation to the order of drops analyzed in the meters. The mean absolute variation in the two devices between capillary samples and PG was significantly higher in individuals with blood glucose levels [ge] 100mg/dL than in individuals with levels [lt] 100mg/dL (p[lt]0.001), but the percentage variation showed no statistical difference (p[gt]0.05). [bold]Conclusions:[/bold] Capillary measurements in the two meters were comparable to PG levels analyzed in the clinical laboratory (variation [lt]10%). Venous samples analyzed in the meters showed satisfactory correlation coefficients, with higher variation in the Optium Xceed device. In conclusion, we observed a good correlation between capillary blood glucose and plasma glucose levels measured by a standard method compared to the two devices tested. Studies to evaluate the correlation of glucometers results and PG levels analyzed by a standard method may improve decision making in diabetes treatment.[br][br]Nothing to Disclose: MGT, SAO, IS, NMS, MER, MN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1487 54 276 SAT-173 PO11-02 Saturday 215 2012


216 ENDO12L_SAT-174 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Cross-Reactivity of Modern Insulin Analogs Insulin Detemir and Insulin Glulisine with Six Automated Insulin Immunoassays Stephen D Merrigan, William E Owen, Sonia L La[apos]ulu, Sara P Wyness, William L Roberts, Joley A Straseski ARUP Institute for Experimental Pathology, Salt Lake City, UT; University of Utah, Salt Lake City, UT Introduction: Identifying whether recombinant insulin analogs cross-react with commercially available insulin immunoassays is critical for accurate result interpretation. While the cross-reactivity of several commonly used analogs has been published previously, it is medically relevant to evaluate newly developed insulin analog formulations across multiple assay platforms.[br]Methods: We determined the cross-reactivity of insulin detemir (Levemir; Novo Nordisk) and insulin glulisine (Apidra; Sanofi-Aventis) with insulin immunoassays on 6 automated platforms at 4 concentrations (30, 100, 300, and 1000 [micro]IU/mL in BSA). Analogs were assayed per manufacturer instructions in duplicate on the Modular E170 (Roche Diagnostics), Liaison (DiaSorin), UniCel DxI 800 (Beckman Coulter), Advia Centaur and Immulite 2000 (Siemens Diagnostics), and Architect i2000 (Abbott Diagnostics) instruments. The ratio of observed and expected analog concentrations was converted to a percent.[br]Results: The Modular E170 and Liaison assays exhibited no cross-reactivity at any concentration with either analog. Cross-reactivity of 1000 [micro]IU/mL insulin detemir exceeded the upper limit of detection of all remaining assays except Immulite 2000 (25%). Conversely, 1000 [micro]IU/mL insulin glulisine exhibited nominal cross-reactivity (maximum 14% by UniCel DxI and Architect i2000). Insulin detemir concentrations from 30 to 300 [micro]IU/mL cross-reacted an average of 17% by UniCel DxI, 52% by Advia Centaur and 22% by Immulite 2000. Insulin glulisine cross-reactivity at these concentrations averaged 6% by UniCel DxI, [lt]3% by Advia Centaur and 8% by Immulite 2000. The most significant cross-reactivity was observed by the Architect i2000 (insulin detemir averaged 101%, with 300 [micro]IU/mL exceeding the upper limit of detection; insulin glulisine averaged 21%).[br]Discussion: This data highlights how exogenous insulins may be detected by, and thus confound the interpretation of, certain insulin immunoassay results. For those assays exhibiting cross-reactivity, insulin detemir was detected at higher percentages than insulin glulisine. All assays utilize two-site (sandwich) antibody detection, thus epitope structure is critical for recognition. Modifications to epitope sites specific to individual analogs may be responsible for the differences observed. Understanding cross-reactivity of automated immunoassays with recombinant analogs is useful for clinical management and diagnosis in patients administering exogenous insulins.[br][br]Sources of Research Support: ARUP Institute for Clinical and Experimental Pathology.[br][br]Nothing to Disclose: SDM, WEO, SLL, SPW, WLR, JAS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2217 54 277 SAT-174 PO11-02 Saturday 216 2012


217 ENDO12L_SAT-175 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) The Impact of Type 2 DM in Mortality of Hospitalized Female Cancer Patients in Taiwan Jen-Der Lin, Wen-Ko Chiou, Feng-Hsuan Liu, Jawl-Shan Hwang, Wei-Ying Chou Chang Gung Memorial Hospital, Chang Gung University, Taoyuan Hsien, Taiwan; Chang Gung University, Taoyuan Hsien, Taiwan [bold]Aim[/bold] The goal of this study was to determine the relationships between diabetes mellitus (DM), specific cancer histologic types (histotypes), age, and mortality in hospitalized women.[br][bold]Methods[/bold] A total of 67,660 females with a mean age 62.76 [plusmn] 14.48 years admitted to the Chang Gung Medical Center in Linkou, Taiwan were diagnosed with cancer or DM. These patients were categorized into the following three groups: patients with cancer, but without DM (group A); patients with cancer and DM (group B); and patients with DM, but without cancer (group C). The disease states of the patients were identified according to the diagnostic codes of the International Classification of Disease-9.[br][bold]Result [/bold]Of the study subjects, 37,204 (54.98 %) were diagnosed with DM. as follows: groups A, n=30,456; group B, n=5,992; and group C, n=31,212. Group B patients had a high frequency of pancreas (34.06%), liver (27.25%), and renal (23.19%) cancers, and a low frequency of thyroid (8.09%), breast (9.30%), and ovarian (12.34%) cancers. Of the 36,448 cancer patients, 2,906 (7.97%) died. The mortality rate in group B (10.46%) was significantly higher than group A and group C. The percentage of DM in patients with different cancers had a negative association with the DM mortality rate/non-DM mortality rate of the corresponding cancer patients.[br][bold]Conclusions[/bold] DM affects the incidence and therapeutic outcomes in female cancer patients. The impact of DM on the mortality rate of specific cancers was negatively associated with the study case of the specific cancer.[br][br]Nothing to Disclose: J-DL, W-KC, F-HL, J-SH, W-YC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 15 54 278 SAT-175 PO11-02 Saturday 217 2012


218 ENDO12L_SAT-176 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Glycemic Variability as Predictor of Clinical Outcome in Insulin-Treated Patients with Type 2 Diabetes Farnoosh Farrokhi, Dawn Smiley, Francisco J Pasquel, Limin Peng, Christopher Newton, Guillermo E Umpierrez Emory University School of Medicine, Atlanta, GA; Rollins School of Public Health, Atlanta, GA Glycemic variability (GV) has been shown to be an independent predictor of mortality in critically ill patients; however, the impact of GV in non-critical patients with T2DM is not known. Accordingly, we determined the ability of GV in predicting hospital complications in a prospective, randomized trial of insulin-treated medical and surgical patients with T2DM. A total of 279 patients with a BG of 140-400 mg/dl were randomized to basal bolus (n=146, age: 59[plusmn]11 yr, admission BG: 210[plusmn]88 mg/dl, A1C: 8.7[plusmn]2.5%, [plusmn]SD), or basal plus supplements (n=133, age: 59[plusmn]13 yr, admission BG: 207[plusmn]83 mg/dl, A1C: 8.3[plusmn]2.3%, [plusmn]SD). Patients in basal bolus were started at 0.5 U/kg, given half as glargine once daily and half as glulisine before meals. Basal Plus received 0.25 U/kg of glargine once daily plus correction doses of glulisine before meals for BG[gt]140 mg/dl. GV was calculated from all BG values using mean delta daily BG and mean standard deviation (SD).[br]Results: Basal plus regimen resulted in similar glycemic control as basal bolus group. Mean daily BG and rate of hypoglycemia after day 1 were 156[plusmn]36 mg/dl and 16% in the basal bolus and 163[plusmn]37 mg/dl and 13% in basal plus group (p=NS). There were no differences in length of stay or in-hospital complications between treatment groups (p=NS). There were no differences in GV between basal bolus and basal plus groups measured by delta or SD (72.5[plusmn]36 vs. 69.3[plusmn]34 mg/dl, and 38.5[plusmn]18 vs. 37.1[plusmn]16 mg/dl, both, p=NS). When GV was assessed in medical and surgical patients, we observed that surgical patients[apos] GV was higher with basal bolus than basal plus (delta: 74.9[plusmn]41 vs. 60.3[plusmn]32 mg/dl, SD: 38.2[plusmn]17 vs. 31.4[plusmn]14 mg/dl, both p=0.02), but there were no differences in GV in medical patients (delta: 70.7[plusmn]32 vs. 76.0[plusmn]34 mg/dl; SD: 38.7[plusmn]17 vs. 41.4[plusmn]16 mg/dl respectively, p=NS). Adopting median GV measures as a cut off, patients with high GV tend to have a higher risk of hypoglycemia compared to those with low GV (SD: 17% vs 7%, p=0.005, delta: 18% vs. 6%, p[lt]0.001). GV was associated with hypoglycemia (p=0.02), but no association was found between GV and hospital complications (p=NS).[br]In summary, treatment with glargine once daily plus correction doses of glulisine before meals resulted in similar glycemic variability, mean daily blood glucose and frequency of hypoglycemia compared to a basal bolus regimen. Higher glycemic variability was associated with increased risk of hypoglycemia but not with hospital complications.[br][br]Disclosures: GEU: Researcher, Sanofi-Aventis. Nothing to Disclose: FF, DS, FJP, LP, CN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2054 54 279 SAT-176 PO11-02 Saturday 218 2012


219 ENDO12L_SAT-177 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) The Impact of Hyperglycemia on Clinical Outcome in Elderly Patients with and without Diabetes in Acute Psychiatric Units: A Multicenter Observational Study Farnoosh Farrokhi, Powell Winter, Isabel Anzola, Pedram Aram, Francisco Pasquel, Shadi Sadeghi-Yarandi, Shoheera Khaliqdina, Charles Gillespie, Limin Peng, Dawn Smiley, Christopher Newton, Guillermo Umpierrez Emory University School of Medicine, Atlanta, GA; Emory University School of Medicine, Atlanta, GA; Rollins School of Public Health, Atlanta, GA The association between acute psychosis, use of antipsychotic therapy and diabetes (DM) is well established; however, the effect of antipsychotic agents on glycemic control and the impact of hyperglycemia on clinical outcome in elderly patients with acute psychiatric illnesses are not known. Accordingly, we analyzed the association between DM and hyperglycemia (BG[gt]140 mg/dl) and clinical outcome in 407 elderly ([gt]60 yrs) subjects (age 77[plusmn]9 yrs, 63% female, BMI 26[plusmn]7 kg/m[sup]2[/sup]) admitted to 2 acute psychiatric units between 1/1/2008 to 12/31/2008. The most common admission diagnoses were acute psychosis (33%), depression (29%), suicidal ideation/attempt (15%) and substance abuse/delirium (23%). A history of DM was present in 127 (31%) patients and 23 (6%) had newly diagnosed hyperglycemia on admission. Compared to nonDM patients, DM had higher BMI (29[plusmn]9 vs 25[plusmn]6 kg/m[sup]2[/sup], p[lt]0.001) and comorbidities: hypertension (87% vs 73%), dyslipidemia (59% vs 38%), and coronary artery disease (31% vs 18%), all p[le]0.002. The admission BG in patients with DM and hyperglycemia (157[plusmn]66 and 162[plusmn]19 mg/dl) was higher than in patients with normoglycemia (101[plusmn]16 mg/dl, p[lt]0.001). There were no differences on admission or daily hospital BG among DM and hyperglycemia patients treated with typical (163[plusmn]67 and 149[plusmn]39 mg/dl), atypical (156[plusmn]60 and 142[plusmn]36 mg/dl) or combination of antipsychotic agents (151[plusmn]75 and 137[plusmn]44 mg/dl), all p=NS. DM patients were treated with diet (9 %), sliding scale regular insulin (SSI, 9 %), oral antidiabetic drugs (OAD) alone (18 %) or in combination with insulin (43%), or basal insulin (22%). A total of 37% had [ge]1 episode of BG [lt]70 mg/dl. Among those with hypoglycemia, 20% were treated with OAD, 44% with OAD and insulin, 31% basal, and 5% with SSI alone. There were no differences in length of stay (LOS) or complications between patients with hyperglycemia or DM compared to those with normoglycemia.[br]In summary, hyperglycemia is a common finding in elderly patients with and without diabetes in acute psychiatric units. Our results indicate that a history of hyperglycemia and DM are not associated with higher risk of hospital complications compared to patients with normoglycemia. In addition, we found no differences in BG concentrations among patients treated with typical, atypical or with combination of antipsychotic agents. Randomized studies are needed to determine the optimal approach to manage hyperglycemia and DM in acute psychiatric disorders.[br][br]Nothing to Disclose: FF, PW, IA, PA, FP, SS-Y, SK, CG, LP, DS, CN, GU 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1922 54 280 SAT-177 PO11-02 Saturday 219 2012


220 ENDO12L_SAT-178 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Serum Sodium Level on Admission Was a Predictor for Mortality in Hyperglycemic Crises: A 5-Year Study in Thailand Pimjai Anthanont, Thana Khawcharoenporn, Thipaporn Tharavanij Faculty of Medicine Thammasat University, Pathum Thani, Thailand; Faculty of Medicine Thammasat University, Pathum Thani, Thailand [bold]Background:[/bold] Hyperglycemic crises, diabetic ketoacidosis (DKA) and hyperosmolar hyperglycemic state (HHS), are the most serious acute complications of diabetes mellitus (DM) which result in significant morbidity and mortality. However, existing and recently updated data on incidence and outcomes of hyperglycemic crises in Southeast Asia are lacking.[br][bold]Methods: [/bold]A retrospective study of DKA and HHS in adults with type 1 or type 2 DM admitted to Thammasat Hospital between 2006 and 2010 was performed. Incidences, precipitating causes, clinical and laboratory characteristics and treatment outcomes of these hyperglycemic crises were obtained via medical record review. Multivariate logistic regression analysis was used to determine predictors for mortality.[br][bold]Results: [/bold]Eighty-three patients were eligible and included. The mean age was 54.9[plusmn]17.7 years old. Most subjects had type 2 DM (85.5%). The 5-year incidence of hyperglycemic crises was 7.46%. Diabetic ketoacidosis occurred more frequently than HHS (4.67% VS 1.71%). During the hyperglycemic episodes, the mean plasma glucose on admission was 741.3[plusmn]320.8 mg/dL. Infections were the most common precipitating factor [61/83 (73.5%)], followed by non-compliance with treatments [35/83 (42.2%)]. Treatment complications included recurrent hyperglycemia (69.9%), hypokalemia (48.2%), hypernatremia (21.7%) and hypoglycemia (15.7%). The overall mortality rate of hyperglycemic crises was 8.4% (5.8% in DKA, 15.8% in HHS and 8.3% in the overlap of both conditions). The most common causes of death were infections [5/7(71.4%)]. By multivariate analysis, serum sodium level on admission was independently associated with mortality (adjusted odd ratio 1.08, 95% CI 1.01-1.16, [italic]P[/italic]= 0.03).[br][bold]Conclusion:[/bold] Hyperglycemic crises were not uncommon in our setting. Diabetic ketoacidosis occurred more frequently but had a lower mortality rate than HHS. Complications from hyperglycemic crisis treatment could be prevented by close monitoring, while high serum sodium level on admission was a predictor for mortality. Strategies to prevent infections and improve treatment compliance are needed to reduce the incidence of hyperglycemic crises among patients with DM.[br][br]Nothing to Disclose: PA, TK, TT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 304 54 281 SAT-178 PO11-02 Saturday 220 2012


221 ENDO12L_SAT-179 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Time to Capillary Blood Glucose Retest in Hypoglycemic Hospitalized Patients Eberta Tan, Richard Chen, Li-Wei Cho, Jen-Min NG Changi General Hospital, Singapore, Singapore Background: Hypoglycemia is common in hospitalised patients with diabetes mellitus due to concomitant illnesses, use of medical therapies or disruption of nutritional intake.(1) Hypoglycemia in hospital can lead to neurological damage, sudden cardiac events and has been associated with higher mortality within 1 year.(2) Prompt recognition and reversal of hypoglycaemia are paramount to prevent such undesirable outcomes. The development of an effective hypoglycaemia prevention and management program requires a good understanding of event patterns in the hospital.[br]Aim: This study intends to assess the time to a capillary blood glucose retest in patients found to be hypoglycemic (defined by capillary blood glucose reading [le]63mg/dl) and if it is affected by whether the patient is in a medical or surgical ward.[br]Methods: A retrospective cross-sectional analysis of all patients in the general ward and intensive care units from 1 to 30 November 2011 was conducted. All patients with capillary blood glucose [le]63mg/dl were identified for analysis. The time from the first capillary blood glucose of [le]63mg/dl to the next biochemical reassessment via capillary blood glucose (time to CGR) was determined for each event. Data showing a time to CGR of more than 2 standard deviations from the mean was excluded from analysis. We analysed time to CGR by ward location. Results are presented as Mean[plusmn]SD and time differences were compared using the two sample T-test.[br]Results: 401 patients were identified for analysis (406 surgical, 671 medical). There were a total of 1077 hypoglycemic events. Collectively, the mean time to CGR was 105[plusmn]86 minutes. There was a significant difference in time to CGR between patients who developed hypoglycaemia in surgical wards and medical wards (117[plusmn]102 versus 98[plusmn]74) (p[lt]0.05).[br]Conclusion: There were more patients who developed hypoglycemia in medical wards compared to surgical wards. Patients who developed hypoglycemia in medical wards were more likely to be retested sooner for biochemical correction of the low blood sugar as compared to patients admitted to surgical wards. Postulated reasons for this include a lack of familiarity with managing diabetes cases and different workloads. Despite these differences, the overall time to correction of hypoglycaemia was still less than ideal. Due to the high prevalence of hypoglycaemia in the acute care setting, it is important to ensure that all hospital care providers are competent in basic diabetes management.[br][br](1) Varghese P, Gleason V, Sorokin R, Senholzi C, Jabbour S, Gottlieb JE. Hypoglycemia in hospitalized patients treated with antihyperglycemic agents. J Hosp Med 2007;2(4):234-40. (2) Turchin A, Matheny ME, Shubina M, Scanlon JV, Greenwood B, Pendergrass ML. Hypoglycemia and clinical outcomes in patients with diabetes hospitalized in the general ward. Diabetes Care 2009;32:1153[ndash]1157.[br][br]Nothing to Disclose: ET, RC, L-WC, J-MN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1891 54 282 SAT-179 PO11-02 Saturday 221 2012


222 ENDO12L_SAT-180 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Survey of Barriers to Effective Inpatient Glucose Control Amita Kathuria, John N Clore, Linda Thurby-Hay Virginia Commonwealth University, Richmond, VA Hyperglycemia in hospitalized patients is a common problem, often contributing to health care costs and outcomes. However, management of hyperglycemia in the hospitalized patient remains a complex matter with multiple factors creating a dynamic state adversely impacting effective glycemic control.[br]The present study was designed to assess current site-specific strategies for inpatient glucose management as determined by a national survey of endocrine fellows.[br]A web-based survey was created using the REDCAP instrument. The survey was then distributed electronically to fellows in endocrinology identified via a listing from the Endocrine Fellows[apos] Foundation, as well as through Association of Program Directors in Endocrinology and Metabolism.[br]A total of 206 responses were received, which is estimated to be about half of the endocrine fellows across the nation. Approximately 53.8% of responses were from large hospitals with 500+ beds. 54.8% of the respondents identified a management protocol at their institution, whereas the remainder (45.1%) reported no management protocol at all. Of those that do not have a standing hospital protocol, strategies for glycemic control included: basal/bolus insulin (61.3%), continuation of home regimen (22.6%), or sliding scale insulin (13.9%). The vast majority of respondents (78.1%) indicated that decisions regarding inpatient glycemic management were heavily dependent upon individual clinical assessment (clinical presentation, prior glycemic control, medical adherence, physician preference). In assessing various barriers impacting inpatient hyperglycemia management, the major impediment appeared to be fear of hypoglycemia, followed by constraints related to timing of meals and procedures in relation to insulin administration. Additionally, while 69.9% of respondents reported obtaining a hemoglobin A1c on inpatients who had not had a recent one in the last 3 months, 30.1% reported no hemoglobin A1c was obtained.[br]In conclusion, the present data suggest that a structured in-hospital glycemic strategy is infrequent. A national randomized control trial is sorely needed and would address a critical factor in hospital outcomes.[br][br]Sources of Research Support: UL 1RR031990.[br][br]Nothing to Disclose: AK, JNC, LT-H 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2311 54 283 SAT-180 PO11-02 Saturday 222 2012


223 ENDO12L_SAT-181 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Frequency of Autoimmune Diseases in Children and Adolescents with Type 1 Diabetes Paulo Cesar Alves Silva, Genoir Simoni, Edson Cechinel, Rose Marie Muller Linhares, Juliana Sande Lee, Ana Carolina Salerno Del Menezzi Tessari, Marilza Leal Nascimento Hospital Infantil Joana de Gusm[atilde]o, Florian[oacute]polis, Brazil; Hospital Universit[aacute]rio da Universidade Federal de Santa Catarina, Florian[oacute]polis, Brazil Introduction: Children and adolescents with type 1 diabetes mellitus (DM1) are at increased risk of developing other autoimmune diseases. Hashimoto[apos]s thyroiditis and celiac disease are the most common disorders found in these patients.[br]Objective: To evaluate the frequency of other autoimmune diseases in outpatients with DM1 treated at Hospital Infantil Joana de Gusm[atilde]o, a reference center in South Brazil.[br]Method: Retrospective study of medical records of 290 patients with DM1 to evaluate the presence of other autoimmune diseases, and this data was analyzed by descriptive statistics.[br]Results: The mean age was 7.4 years old, 52.4% were male. Other autoimmune diseases were observed in 16.5% of cases. The most frequent was Hashimoto[apos]s thyroiditis with 9.3%, followed by celiac disease with 2.4%. There was one case of each of the following diseases: Graves[apos] disease, psoriasis and vitiligo.[br]Conclusion: The high frequency of autoimmune diseases associated with DM1 emphasizes the importance of attention to the signs and symptoms of these diseases and screening for adequate control of these patients.[br][br]Nothing to Disclose: PCAS, GS, EC, RMML, JSL, ACSDMT, MLN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 336 54 284 SAT-181 PO11-02 Saturday 223 2012


224 ENDO12L_SAT-182 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Hemoglobin A1c: The New Gold Standard in Type 2 DM Screening? Dawn M Weiler, Leonie Sutherland, Shawn R Simonson, Jonathan Glogowski Boise State University, Boise, ID; Boise State University, Boise, ID; Duke Children[apos]s Hospital [amp] Medical Center, Durham, NC [bold]Purpose:[/bold] Compare traditional fasting plasma glucose (FPG) and the recently published hemoglobin A1c (HbA1c) diagnostic criteria in screening for type 2 diabetes in a population of Latino adults.[br][bold]Background:[/bold] Diabetes is one of the fastest growing disease classifications within the United States; especially among the Latino population. Review of the literature reveals that up to 60% of individuals have micro and macro-vascular disease related complications present at the time of type 2 diabetes diagnosis. Additionally, it is estimated that for many the disease has been present for as long as seven years before formal diagnosis. Early diagnosis and effective treatment significantly reduces the long-term complications and costs often associated with diabetes. These numbers validate that aggressive, cost effective, and efficient screening methods are imperative for improved health outcomes.[br][bold]Methods:[/bold] A descriptive study was carried out with 225 Latino adults (female n=132; male n=93) ranging in age from 18-75 years (m=43). Demographic, health behavior (Health Promoting Lifestyle Profile II), anthropometric (height, mass, waist-hip ratio, Jackson-Pollock 3 site skinfolds), and fasting venous blood samples (lipid, insulin, glucose, HbA1c) were collected. Diabetes status was assessed utilizing traditional FPG and newly released HbA1c guidelines. Participants self-reporting a known diabetes diagnosis (n=27) were excluded from the analysis.[br][bold]Results:[/bold] Two and a half percent (n=5) met diabetes diagnosis criteria utilizing FPG compared to 7.6% (n=15) based on HbA1c criteria. Fifty-seven participants (28.9%) had HbA1cs between 5.6 and 6.4 of whom 56% (n=32) had normal FPG values. Of those with HbA1cs between 6.0 and 6.4 (n=22), 32% (n=7) had normal FPG levels. In comparison, of the 27% (n=53) who met [ldquo]impaired fasting glucose[rdquo] criteria, 10.2% (n=20) had normal (5.5 and lower) HbA1c values.[br][bold]Conclusions:[/bold] In this population, FPG had a 25% false negative and a 37.7% false positive screening rate when compared to HbA1c values. Fasting plasma glucose also failed to identify 66.7% of individuals who met HbA1c criteria for diabetes diagnosis. These results imply that utilizing HbA1c for type 2 diabetes screening is a more sensitive diagnostic tool and should become the new [ldquo]gold standard[rdquo] due to its increased accuracy, efficiency and lower overall cost (personal and financial) compared to traditional FPG methods.[br][br]Sources of Research Support: Boise State University School of Nursing Faculty Research Grant; Boise State University College of Health Sciences Mini-Research Grant; Boise State University Office of Sponsored Program Grant.[br][br]Nothing to Disclose: DMW, LS, SRS, JG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 625 54 285 SAT-182 PO11-02 Saturday 224 2012


225 ENDO12L_SAT-183 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) The Well-Being and Treatment Satisfaction of Diabetic Patients in General Hospital in Korea Ie Byung Park, Sin Na Lee, Young Sil Eom, Sei-Hyun Kim, Ki Young Lee, Sihoon Lee, Yeun Sun Kim Gachon University Gil Hospital, Incheon, Republic of Korea [bold]Backround[/bold][br]The ambulatory diabetes care system in Korea is different from those in America. In Korea, the number of patients visited per day is about 80-160 diabetic patients, and the treatment time per patient is so short(3-5 minutes). We investigated whether the well-being and treatment satisfaction of diabetes in general hospital in Korea are different from those in westernized ambulatory care. We analysed relationship between well-being and treatment satisfaction and affecting factors on both.[br][bold]Methods[/bold][br]This study subjects were 382 diabetics, 20-80 years old, who visited the Department of Endocrinology at Gachon University Gil Hospital. We evaluated the well-being using 12-item well-being questionnaire(WBQ-12) consisting of three 4-item subscales(Negative Well-Being(NWB), Energy(ENE), and Positive Well-Being(PWB), and 12-item overall scale General Well-Being(GWB)). We evaluated the diabetes treatment satisfaction using Diabetes Treatment Satisfaction Questionnaire(DTSQ).[br][bold]Results[/bold][br]Cronbach[apos]s alpha for WBQ-12 and DTSQ were 0.819 and 0.819, respectively. The higher total WBQ-12(GWB) score was significantly associated with women(r=0.20), educational status(r=0.15), income(r=0.30), satisfaction for waiting time(r=0.20), exercise compliance(r=0.21), total pill number(r=-0.16) and HbA1c level(r=-0.15). The NWB, ENE, and GWB score in men is significantly different from those in women(2.16[plusmn]2.3 vs 3.89[plusmn]3.3; 6.69[plusmn]2.5 vs 5.90[plusmn]2.8; 23.25[plusmn]6.5 vs 20.56[plusmn]7.0, respectively). ENE score in men was lower than that in women(5.56[plusmn]2.8 vs 6.56[plusmn]2.6, p=0.01) and ENE score in insulin-user was lower than non-user(5.56[plusmn]2.8 vs 6.56[plusmn]2.6. p=0.01).[br]The higher DTSQ score was significantly associated with age(r=0.24), education(r=-0.12), waiting time for treatment(r=0.34), satisfaction of treatment time(r=0.60), diet compliance(r=0.42) and exercise compliance(r=0.35).[br]In multiple regression analysis, GWB score was significantly associated with income and satisfaction of treatment time and DTSQ score was significantly associated with diabetes duration, satisfaction of treatment time and diet compliance.[br][bold]Conclusion[/bold][br]Our results show that some affecting factors on the well-being and diabetes treatment satisfaction in ambulatory diabetic patients in Korea are similar to those in westernized outpatient clinics. But the environmental conditions such as treatment time and waiting time for treatment are likely to be important factors affecting on those scores compared with those in Europe and America.[br][br]Nothing to Disclose: IBP, SNL, YSE, S-HK, KYL, SL, YSK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1170 54 286 SAT-183 PO11-02 Saturday 225 2012


226 ENDO12L_SAT-184 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Gestational Diabetes in Assisted Reproductive Techniques and Spontaneous Pregnancy Radhika Jindal, Nitin Gupta, Mukul Gupta, Mohammad Asim Siddiqui, Subhash Kumar Wangnoo Indraprastha Apollo Hospital, New Delhi, India OBJECTIVE:[br]The aim of this study was to compare the maternal glycemia, weight gain and fetal outcomes in women diagnosed with Gestational Diabetes Mellitus (GDM) during pregnancy with Assisted Reproductive Techniques (ART) and spontaneous pregnancies.[br]METHODS:[br]Thirty-six women who had successful ART (irrespective of indication) and developed GDM during the course of their singleton pregnancy (ART group) and 37 non-IVF women with GDM (non-ART group) were included in the study after informed consent. They were matched according to age and pre-pregnancy BMI. None had any previous history of diabetes mellitus or GDM before their pregnancies. GDM was diagnosed as per the ADA criteria for diagnosis of GDM. The patients in both the groups received individualized care from the diabetes educators and the treating endocrinologists. We compared the maternal characteristics, course of pregnancy and neonatal outcome.[br]RESULTS:[br]The weight gain until the diagnosis of GDM in both non-ART and ART groups of women was not significantly different (9.51[plusmn]3.97 vs 10.0[plusmn]3.8kg, p=0.8 respectively). First trimester fasting glucose levels were found to be significantly higher in ART group (88[plusmn]15.2 vs 81[plusmn]10.3mg/dl, p=0.04). Second trimester oral glucose tolerance test (OGTT) results and glucose levels during GDM treatment did not differ between the groups. There were no significant changes in investigated fetal and neonatal parameters: 3rd trimester abdominal circumference, percentile and neonatal birth weight (3250[plusmn]641 vs 2900[plusmn]440g, p=0.22).[br]CONCLUSIONS:[br]GDM among women who underwent ART is associated with higher first trimester fasting glucose levels. We suggest that women undergoing ART be evaluated for dysglycemia at an earlier gestational age.[br][br]Nothing to Disclose: RJ, NG, MG, MAS, SKW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1460 54 287 SAT-184 PO11-02 Saturday 226 2012


227 ENDO12L_SAT-185 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Pre-IGT in Pregnancy: Comparison of HbA1c and Clinical Scoring in Patients with Gestational Diabetes (GDM) Mary Jane Yu Tanchee-Ngo, Leilani B Mercado-Asis University of Sto Tomas Hospital, Manila, Philippines Background: In the initial stage of Diabetes mellitus, the [beta]-cell compensates for the insulin resistance by increasing insulin secretion to maintain normoglycemia, which we termed pre-impaired glucose tolerance (pre-IGT state). The 2nd hour glucose is normal ([lt]140mg/dl) but with increased insulin levels ([gt] 30uIU/ml) after 75 grams oral glucose tolerance test (OGTT)1,2. Clinical scoring developed by us for non-pregnant subjects using risk factors of diabetes mellitus showed similar scores in normal and pre-IGT nonpregnant individuals implying that the latter is a subclinical condition2. Although our group demonstrated that a 2nd hr insulin level of 45 uIU after 75 grams OGTT done during the first trimester is predictive of development of GDM3, elucidation of pre-IGT among pregnant individuals with risk factors has yet to be done.[br]Objective: To identify pre-IGT among pregnant individuals with risk factors for GDM.[br]Methodology: This is an on-going retrospective cohort study of pregnant individuals 18-40 year-old seen at the Endocrine Clinic of our institution. Risk factors for diabetes were determined. All underwent Second-hour blood glucose and insulin after 75 g oral glucose tolerance test (OGTT) with glycosylated hemoglobin (HbA1c). They were then divided into 4 groups base on their glycemic status and insulin levels.: Group A (normoglycemia with normal insulin levels), Group B or the Pre-IGT group (normoglycemia with high insulin levels and Group C or the IGT and GDM group (hyperglycemia with high insulin levels). The clinical risk scoring of the normoglycemic group was then determined. The most frequent risk factor given the highest score and the less frequent risk factor given the lowest score. A p value equal or less than 0.05 was considered significant.[br]Results: A total of 11 out of 16 patients fulfilled the criteria. Thirty percent were found to have GDM with hyperinsulinemia whereas 70% were found to have pre-IGT. Although both insulin levels were increased, the value in the GDM group was not significantly different in the pre-IGT group (236 uIU vs. 121 uIU, p=0.053). The most frequent risk factors found in both groups is a family history of DM followed by an elevated BMI. There was no significant difference between the mean clinical risk scores of the two groups (p 0.62).[br]Conclusion: Preliminary observations showed pre-IGT exists among high-risk pregnant subjects whereas clinical risk scoring of GDM and pre-IGT patients did not vary.[br][br]1. Matawaran B, Mercado-Asis L. Comparison of Pancreatic Insulin Response to Hyperglycemia Among Filipino Subjects of Various Glycemic Status. Phil J Internal Medicine, 47: 25-30, Jan-Feb 2009. 2. Nisce I, Matawaran B, Mercado-Asis L. Clinical Risk Scoring is Not Useful in Screening Patients with Pre-IGT (Hyperinsulinemic State) : The Importance of 2nd-hour Insulin Measurement in OGTT Poster in PSEM 2011, EndoSoc USA 2011. 3. Quiambao D., Fonte, J., Mercado-Asis L., Matawaran B., AMORADO-SANTOS, F., Lejos, A., Cabatu, M., Decena, C., Dee, M., Gamilla, Z., Gutierrez, R., Hyperinsulinemia During the First Trimester as a Predictor in the Development of Gestational Diabetes Mellitus Among High Risk Pregnant Patients. 4. Standards of Medical Care in Diabetes-2011. January 2011 vol. 34 no. Supplement 1 S11-S61.[br][br]Nothing to Disclose: MJYT-N, LBM-A 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2252 54 288 SAT-185 PO11-02 Saturday 227 2012


228 ENDO12L_SAT-186 POSTER SESSION: Diabetes [amp] Clinical Care (1:30 PM-3:30 PM) Postpartum Women Have Less Insulin Resistance and Better Glucose Tolerance Compared with Pregnant and Non-Pregnant Women David Gustavo Garcia-Gutierrez, Ma Ludivina Robles-Osorio, Pablo Garcia-Solis, Juan Carlos Solis-S, Hebert Luis Hernandez-Montiel Facultad de Medicina, Universidad Aut[oacute]noma de Quer[eacute]taro, Quer[eacute]taro, Mexico INTRODUCTION:[br]Pregnancy is a period during which there are changes such as insulin resistance and more frequent glucose intolerance and gestational diabetes. The study of this conditions and its changes during postpartum are important due to the group of patients who develop important changes during pregnancy, such as glucose intolerance and gestational diabetes than are prognostic factors to develop glucose intolerance and type 2 diabetes in the future.[br]METHODS:[br]Cross sectional study, we studied 304 women, mean age was 26.8[plusmn]5.3 years old; 192 pregnant, 47 women were in postpartum (between 8 to 12 weeks after delivery), age 26.3[plusmn]5.3 years old; 65 non-pregnant women, mean age 28.7[plusmn]5-8 years old. Glucose and insulin were measured on fasting and 1, 2 and 3 hrs. after an oral glucose load.[br]RESULTS:[br]We investigated the weight before pregnancy and current weight in no pregnant women, we found 30.6% overweight women and 15.1% obesity, both diagnosis were present in 45.7%. According to the Institute of Medicine 21.9% of pregnant women had increased more weight per trimester than recommended.[br]We found that fasting glucose was different between first [italic]vs[/italic] second (p[lt]0.001) and third trimesters (p=0.012), a significant difference was found only in the first hour between first and third trimester (p=0.05), but not difference was found within the other groups regarding to glucose and insulin levels. When we compared the glucose area under the curve (AUC) we found a significant difference between the groups (p[lt]0.001); being the higher glucose and insulin AUC within the third trimester pregnant women, and the lowest AUC for glucose and insulin in the postpartum group, even compared with the non-pregnant women.[br]Between the pregnant women there was no difference within the different trimesters on insulin and glucose levels.[br]CONCLUSIONS:[br]We found in a population with a high prevalence of diabetes and metabolic syndrome, a significant difference between the pregnant women and the postpartum (8 to 12 weeks after delivery), with glucose and insulin lower levels even comparing with non-pregnant women, this finding can be related to metabolic changes given after pregnancy, after a state of significant insulin resistance and a increase in glucose levels during the OGTT. The study of this changes can be of physiological interest and also in order to identify which factors also can contribute to a good recovery after pregnancy and contribute to develop type 2 diabetes in the future.[br][br]Sources of Research Support: Programa para el mejoramiento del profesorado, secretaria de educacion publica. (PROMEP).[br][br]Nothing to Disclose: DGG-G, MLR-O, PG-S, JCS-S, HLH-M 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2347 54 289 SAT-186 PO11-02 Saturday 228 2012


229 ENDO12L_SAT-187 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Hsp60, a Leptin-Induced Mitochondrial Chaperone, Impacts on Central Insulin Signaling Andre Kleinridders, Siegfried Ussar, Jane Hvarregaard Christensen, Marcelo Mori, Peter Bross, C Ronald Kahn Joslin Diabetes Center, Boston, MA; Aarhus University Hospital and Aarhus University, Aarhus, Denmark; Aarhus University, Aarhus, Denmark The adipocyte-derived hormone leptin is secreted after food consumption and acts on the hypothalamus to control energy metabolism by increasing energy expenditure and promoting satiety. Heat shock protein (Hsp) 60, a nuclear encoded mitochondrial chaperone, is crucial for proper folding of mitochondrial proteins and plays a pivotal role in maintaining mitochondrial function and cell homeostasis. Here we show a 50% decrease in the level of Hsp60 in the hypothalamus at both the mRNA and protein level of leptin-resistant db/db mice and leptin-deficient ob/ob mice. This is due to direct regulation of Hsp60 expression by leptin via activation of STAT3 as overexpression of a dominant negative STAT3 abolishes leptin[apos]s ability to induce Hsp60 expression. Consistent with this, intraperitoneal injection of leptin in normal mice led to an upregulation of Hsp60 in the hypothalamus [italic]in vivo[/italic].[br]As Hsp60 is crucial for the integrity of the mitochondrial matrix, we assessed mitochondrial respiration using isolated mitochondria from hypothalamic samples of db/db mice in a Seahorse Flux Analyzer. When compared to control mitochondria, mitochondria from db/db mice exhibited a defect in the electron transport chain activity. This defect was preserved [italic]in vitro[/italic] by lentiviral knockdown of Hsp60 in the hypothalamic cell line N25/2. This was associated with decreased levels of multiple protein subunits of complexes I, II, III and V. As a consequence of this, reactive oxygen species production was increased as assessed by measuring lipid peroxidation. In addition, the mitochondrial dysfunction produced by knockdown of Hsp60 [italic]in vitro[/italic] led to activation of c-Jun kinase pathway, increased IRS1 Ser307 phosphorylation, and resistance to normal insulin signaling, which could be restored by inhibiting reactive oxygen species using N-acetyl-L-cysteine. Consistent with the insulin resistance observed [italic]in vitro[/italic], mice with a haploinsufficiency of Hsp60 showed insulin resistance in the hypothalamus due to mitochondrial dysfunction specifically in the hypothalamus.[br]Together these data show that leptin plays an important role in regulating mitochondrial function in the hypothalamus by regulating the mitochondrial chaperone Hsp60 which influences insulin sensitivity in the hypothalamus. These data provide a novel pathway of leptin/insulin crosstalk in the central nervous system which may impact on hypothalamic control of energy homeostasis in obesity and insulin resistant states.[br][br]Nothing to Disclose: AK, SU, JHC, MM, PB, CRK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 930 55 290 SAT-187 PO14-02 Saturday 229 2012


230 ENDO12L_SAT-188 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Insulin Action on Brain Affects Serotonergic and Dopaminergic Neurotransmission and Mediates Behavioral Changes with Aging Andre Kleinridders, Laura Cappellucci, Sara Vienberg, Emmanuel N Pothos, C Ronald Kahn Joslin Diabetes Center, Boston, MA; Tufts University School of Medicine, Boston, MA Insulin is secreted from the pancreas in response to rise in blood glucose and acts on peripheral tissues to control glucose and lipid metabolism. Insulin also acts on neurons in the brain in control of energy homeostasis and feeding behavior. We have previously shown that mice with a disruption of the insulin receptor in the brain (NIRKO mice) have increased food intake leading to some features of metabolic syndrome. In addition, these mice develop hypothalamic hypogonadism, but have no detectable behavioral abnormalities at a young age. Here we show that NIRKO mice subjected to aging develop significant alterations in the dopaminergic and serotonergic systems leading to increased anxiety and decreased exploratory drive, as well as impaired motor coordination.[br]To test anxiety and explorative behavior, aging NIRKO mice were subjected to mouse tail suspension, forced swimming and adapted open field tests. By 17 months of age, all of these tests showed significantly increased periods of immobility indicative of increased anxiety, decreased explorative drive and depressive-like behavior. These changes were not apparent in the 11 month old mice. Assessment of motor skills in the older cohort using the balance beam also revealed impairment of NIRKO mice in motor balance and coordination.[br]To understand the cause of these phenomena, we examined neurotransmitter receptor expression and neurotransmitter release in control and NIRKO mice with aging. Dopamine release in the mesolimbic system assessed using carbon fiber amperometry displayed a significant decrease in amplitude and width of the dopamine peak, as well as a decreased half-life of dopamine in the nucleus accumbens, which was independent of neurotransmitter reuptake and not altered by the reuptake inhibitor nomifensine. This phenotype was already observed at age of 8 month where no behavioural abnormalities were detected. Furthermore, even in young (3 month old) NIRKO mice, we could observe decreased expression of some serotonin receptors (Htr1B, Htr2A and Htr7) in the hippocampus of NIRKO mice. Insulin treatment of cultured neuronal cells in vitro showed that these serotonin receptors were directly induced by insulin treatment.[br]Thus, taken together these data show that insulin signaling can act directly on neurons to alter both serotonergic and dopaminergic signaling. While initially these alterations are clinically silent, with aging, these alterations results in changes in behavior and motor skills.[br][br]Nothing to Disclose: AK, LC, SV, ENP, CRK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1998 55 291 SAT-188 PO14-02 Saturday 230 2012


231 ENDO12L_SAT-189 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Novel Role of Insulin and IGF-1 Signaling in Regulation of DNA Imprinting Jeremie Boucher, Marika Charalambous, Kim Zarse, Marcelo Mori, Michael Ristow, Anne Ferguson-Smith, C Ronald Kahn Joslin Diabetes Center, Boston, MA; University of Cambridge, Cambridge, UK; University of Jena, Jena, Germany Insulin and insulin like growth factor 1 (IGF-1) control many biological processes including cellular metabolism, proliferation, differentiation and apoptosis. Insulin and IGF-1 receptors (IR and IGF1R) also create novel signals in the unoccupied state, such that preadipocytes lacking both insulin and IGF-1 receptors are resistant to apoptosis. Double knockout cells (DKO) also exhibit major changes in miRNA regulation. Among the 371 miRNAs detectable in preadipocytes, 134 were differentially expressed between control and DKO cells (p[lt]0.05) and 89 of them were down-regulated in DKO cells compared to controls, in many cases by 10- to [gt]1000-fold. This occurred without any changes in expression of proteins involved in miRNA maturation and processing such as Dicer, Drosha, DGCR8, TRBP, Ago2 and Exportin 5. Most of these down-regulated miRNAs reside in two miRNA clusters on mouse chromosomes 2 and 12 previously shown to be regulated by imprinting. Inactivation of the IR alone in mouse embryonic fibroblasts also resulted in a down-regulation of many of these imprinted miRNAs. In DKO cells, the imprinted genes in these loci, Dlk1, Gtl2, Rtl1, Dio3 and Sfmbt2, whether normally maternally or paternally expressed, along with several imprinted genes on other chromosomes, such as IGF2, H19 or Phlda2 were highly down-regulated or undetectable, indicating generalized epigenetic misregulation of imprinted genes. Pyrosequencing of bisulfite-treated DNA from the DKO preadipocytes revealed increased methylation of several differentially methylated regions, such as the Dlk1-DMR, IG-DMR, Gtl2 promoter or the H19 DMD, which occurred without changes in expression in DNA methyl transferases 1, 3a or 3b. Treatment of DKO cells with a methyltransferase inhibitor partially reversed the changes in miRNA and gene expression. Increased methylation correlated with a [gt]70% decrease in expression of the transcription factor Zac1, which has been shown to regulate a large imprinted network. Thus, disruption of insulin and IGF-1 signaling in somatic cells can induce changes in the epigenome, causing a down-regulation in expression of imprinted genes, at least in part, due to down-regulation of Zac1 and increased DNA methylation.[br][br]Nothing to Disclose: JB, MC, KZ, MM, MR, AF-S, CRK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2189 55 292 SAT-189 PO14-02 Saturday 231 2012


232 ENDO12L_SAT-190 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Attenuation of High Glucose-Induced Oxidative Stress in Brain Pericytes by Ethoxyzolamide, a Mitochondrial Carbonic Anhydrase Inhibitor Tulin O Price, Nader Sheibani, Ping Patrick, Nuran Ercal, Gul N Shah Saint Louis University, St Louis, MO; Missouri University Science[amp]Technology, Rolla, MO; University of Wisconsin, Madison, WI Diabetes mellitus has strongly been associated with the imbalance between production of reactive oxygen species (ROS) and the antioxidant enzyme system. Overproduction of mitochondrial ROS results in oxidative stress which is augmented during diabetes in insulin-insensitive tissues such as brain. Pericytes (PC) in the microvasculature of brain are especially susceptible to oxidative stress. Mitochondrial carbonic anhydrases CA VA and CA VB (mCAs) regulate oxidative metabolism of glucose, and thus, may play an important role in the ROS generation and oxidative stress. We have recently shown that inhibition of mCAs reduces diabetes-induced oxidative stress in the mouse brain and rescues cerebral PC loss. Here we tested the hypothesis that pharmacological inhibition of mCAs attenuates high glucose-induced intracellular oxidative stress in brain PC.[br]We have isolated conditionally immortalized cerebral PC from transgenic Immorto mouse. These cells were analyzed for primary PC markers by fluorescence[ndash]activating cell sorting (FACS) and for the presence of mCAs by RT-PCR and Western blotting. These cells were grown in low glucose (LG, 5.7 mM) and high glucose (HG, 40.7 mM) for 6 days. The measures of oxidative stress were reduced glutathione (GSH), and activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR). For pharmacological inhibition of mCAs, ethoxyzolamide (ETZ) was added to the culture medium from day one of the experiment.[br]The PC expressed PDGFR-[beta], NG2, CD13, and [alpha]-SMA markers. PECAM-1, the endothelial cell marker was not expressed. These cells were successfully passaged and maintained in culture for several months without loss of these markers. Six days exposure to high glucose resulted in significant decrease in GSH (100[plusmn]5.4 LG, 39.95[plusmn]3.18 HG, p[lt]0.0007), SOD (100[plusmn]17.1 LG, 41.58[plusmn]7.44 HG, p[lt]0.03), and GR (100[plusmn]9.86 LG, 82.64[plusmn]1.49 HG, p[lt]0.05) and an increase in GPx (100[plusmn]13.02 LG, 130.3[plusmn]6.65 HG, p[lt]0.05) activities compared to low glucose indicating increased intracellular oxidative stress. Treatment of PC with ETZ significantly reduced high glucose-induced oxidative stress.[br]These results provide the first evidence that high glucose induces oxidative stress in cultured mouse brain PC, and pharmacologic inhibition of mCAs attenuates high glucose-induced oxidative stress.[br][br]Nothing to Disclose: TOP, NS, PP, NE, GNS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1804 55 293 SAT-190 PO14-02 Saturday 232 2012


233 ENDO12L_SAT-191 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Mitochondrial Carbonic Anhydrase Inhibition Protects Kidneys from Hyperglycemia-Induced Oxidative Damage in Diabetic Mice Uma Muthyala, Tulin Price, Ping Patrick, Gul Shah Saint Louis University, St Louis, MO [bold]Introduction[/bold]: Diabetes mellitus (DM) is the leading cause of end-stage renal disease.Increased mitochondrial oxidative stress (OxSt) mediated by reactive oxygen species (ROS) is implicated in hyperglycemia-induced kidney dysfunction.[br][bold]Objective[/bold]: We hypothesize that inhibition of mitochondrial carbonic anhydrases (mCA), regulators of oxidative metabolism of glucose, reduces OxSt in the kidneys and helps slow down the onset and progression of diabetic nephropathy (DN).[br][bold]Methods[/bold]: We used mCA double knockout (DKO) and diabetic mice to test our hypothesis. Diabetes was induced by streptozotocin and pharmacological inhibition of mCA was accomplished by CA inhibitor, topiramate(TOP), administered daily for 12 weeks. Urine samples were analyzed for albumin and creatinine. Kidneys were extracted for determination of OxSt.[br][bold]Results[/bold]: Diabetes-induced OxSt indicated by decreased reduced glutathione (GSH, 100 [plusmn] 9.22 control, 71.89 [plusmn] 3.82 DM, 107.1 [plusmn] 9.06 DM+TOP, p [lt] 0.02) and increased 4-hydroxy-2-trans-nonenal (HNE, 100 [plusmn] 17.66 control, 607.1 [plusmn] 176.6 DM, 290.8 [plusmn] 52.02 DM+TOP, p [lt]0.02) was reversed with TOP treatment. An elevated albumin/creatinine ratio in DM (100.93 [plusmn] 9.43 g/mg) was reduced upon TOP treatment (73.91 [plusmn] 5.43 g/mg, p[lt]0.05) as well. The superoxide dismutase(SOD) activity remained unchanged in DM and DM+TOP mice. The above results were supported by DKO data.[br][bold]Discussion[/bold]: The unchanged SOD activity in DM and DM+TOP mice suggests that mCA inhibition reduces OxSt and albuminuria by decreasing the production of ROS rather than elimination of ROS by increasing SOD activity.[br][bold]Conclusions[/bold]: Mitochondrial CA may provide a novel therapeutic target for slowing down the onset and progression of DN.[br][br](1)Brownlee M. The pathobiology of diabetic complications: a unifying mechanism. Diabetes 2005;54(6). (2)Saito Y, Kida H, Takeda S, Yoshimura M, Yokoyama H, Koshino Y, Hattori N. Mesangiolysis in diabetic glomeruli: its role in the formation of nodular lesions. Kidney Int. 1988 Sep;34(3):389-96. (3)Kriz W, Lemley KV. The role of the podocyte in glomerulosclerosis. Curr.Opin.Nephrol.Hypertens. 1999 Jul;8(4):489-97.[br][br]Nothing to Disclose: UM, TP, PP, GS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 160 55 294 SAT-191 PO14-02 Saturday 233 2012


234 ENDO12L_SAT-192 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) [D-Leu-4]-OB3, a Synthetic Peptide Amide with Leptin-Like Activity, Enhances the Effects of Orally Delivered Exenitide (Byetta[reg]) and Pramlinitide Acetate (Symlin[reg]) on Body Weight Gain and Glycemic Control in Leptin-Resistant Male C57BLK/6-m [italic]db/db[/italic] Mice Matthew C Leinung, Patricia Grasso Albany Medical College, Albany, NY In the present study, exenitide (Byetta[reg]) or pramlinitide acetate (Symlin[reg]), currently administered by subcutaneous injection, were reconstituted in Intravail[reg], a patented transmucosal absorption enhancing agent, in the absence or presence of [D-Leu-4]-OB3, and delivered orally by gavage to leptin-resistant male C57BLK/6-m [italic]db/db[/italic] mice twice daily for 14 days. Body weight gain, food and water intake, blood glucose, and serum insulin levels were measured. Mice receiving Intravail[reg] alone for 14 days were 21.1% heavier at the end of the test period, whereas oral delivery of Byetta[reg] or [D-Leu-4]-OB3 in Intravail[reg] resulted in significantly less body weight gain, 13.9% and 11.5%, respectively. Most worthy of note, mice receiving Byetta[reg] and [D-Leu-4]-OB3 were 2.2% lighter than they were at the beginning of the study. Similar results were seen with Symlin[reg]. In another study, mice receiving Symlin[reg] or [D-Leu-4]-OB3 in Intravail[reg] were 26.8% and 25.4% heavier, respectively, after 14 days, while Intravail[reg] treated control mice were 38.2% heavier than their initial body weight. Co-administration of Symlin[reg] and [D-Leu-4]-OB3 did not significantly enhance the effect of Symlin[reg] on body weight gain. Food intake was significantly reduced by Byetta[reg], Symlin[reg] and [D-leu-4]-OB3 alone, and co-delivery of Byetta[reg] or Symlin[reg] with [D-Leu-4]-OB3 did not induce any further reduction. Oral delivery of Byetta[reg] or Symlin[reg] in Intravail[reg] significantly lowered blood glucose levels by 20.4% and 22.0%, respectively, and co-delivery of Byetta[reg] or Symlin[reg] with [D-Leu-4]-OB3 further reduced blood glucose by 32.3% and 40.0% respectively, of initial levels. A dose-related increase in serum insulin was observed in response to increasing concentrations of Byetta[reg], and [D-Leu-4]-OB3 enhanced the insulin response to Byetta[reg] at all concentrations tested. No similar increases in serum insulin occurred in response to increasing concentrations of Symlin[reg]. The results of these studies indicate that (1) oral delivery of Byetta[reg] and Symlin[reg] is feasible; (2) the bioactivity of Byetta[reg] and Symlin[reg] is retained following oral delivery in Intravail[reg]; and (3) the effects of Byetta[reg] and Symlin[reg] on energy balance and glycemic control can be enhanced by co-administration with [D-Leu-4]-OB3, a synthetic peptide amide with leptin-like activity.[br][br]Sources of Research Support: Willard B. Warring Memorial Fund. Aegis Therapeutics, San Diego, CA.[br][br]Nothing to Disclose: MCL, PG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 133 55 295 SAT-192 PO14-02 Saturday 234 2012


235 ENDO12L_SAT-193 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) CNS Vitamin D Improves Glucose but Not Insulin Tolerance in Diet-Induced Obesity Stephanie R Sisley, Randy J Seeley, Darleen A Sandoval Cincinnati Children[apos]s Hospital Medical Center, Cincinnati, OH; University of Cincinnati, Cincinnati, OH Vitamin D deficiency is associated with obesity and type 2 diabetes, although the mechanism is unknown. Multiple lines of evidence support the CNS activity of the NF[kappa]B pathway as a link in the glucose dysregulation accompanying diet-induced obesity. Given that vitamin D receptors are present in the hypothalamus, the key CNS region for weight and glucose regulation, we sought to determine if acute downregulation of this pathway in the hypothalamus with 1,25-hydroxyvitamin D, an IKK[beta] inhibitor in the NF[kappa]B pathway, decreases food intake or improves glucose tolerance. No effects on food intake were observed after direct administration of 100[mu]g 1,25-hydroxyvitamin D or vehicle (1[mu]l) into the third cerebral ventricle (i3vt) of diet-induced obese Long-Evans rats 60 minutes prior to the dark cycle. We then performed an i3vt injection of 1,25-hydroxyvitamin D 60 minutes prior to an intraperitoneal glucose tolerance test (ipGTT). I3vt 1,25-hydroxyvitamin D significantly decreased the glucose excursion 15 and 30 minutes after the glucose load compared to vehicle. Interestingly, no significant differences in glucose levels were seen during an intraperitoneal insulin tolerance test after i3vt injection of 1,25-hydroxyvitamin D compared to vehicle. We speculate that the positive ipGTT results are secondary to an improved first phase insulin response. Therefore, acute inhibition of NF[kappa]B by vitamin D, specifically within the CNS, improves glucose tolerance. However, the mechanism of improvement is not secondary to improved insulin sensitivity.[br][br]Sources of Research Support: NIH Grant 5T32ES010957-1 awarded to SRS; Fellows Development Research Grant Program in Diabetes, Obesity and Fat Cell Biology awarded to SRS.[br][br]Disclosures: DAS: Research Funding, Johnson & Johnson, Novo Nordisk, Pfizer, Inc; Speaker, Novo Nordisk. Nothing to Disclose: SRS, RJS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1953 55 296 SAT-193 PO14-02 Saturday 235 2012


236 ENDO12L_SAT-194 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Physiological Effect of Peripheral Serotonin on Glucose and Lipid Metabolism in Mice and Sheep Hitoshi Watanabe, Ryo Saito, Tatsuya Nakano, Hideyuki Takahashi, Yu Takahashi, Keisuke Sumiyoshi, Yuya Nagasawa, Shunsuke Terada, Natsumi Okada, Dian W Harjanti, Natsumi Sekiguchi, Hiroaki Sano, Kouichi Watanabe, Hisashi Aso Tohoku University, Sendai, Japan; Iwate University, Morioka, Japan [Objective][br]Serotonin is a monoaminergic neurotransmitter with activities that modulate central and peripheral functions. Serotonin affects food intake, sleep, anxiety, sexual behavior and mood in the central nervous system. On the other hand, the functions of serotonin in peripheral tissue have not yet been fully elucidated. Further, serotonin is thought not to be able to pass the blood-brain barrier. Thus, there are two independent serotonin systems: one in the central nervous system and the other in the periphery. Recently, peripheral serotonin is known to be associated with glucose metabolism mainly because of its regulation of the secretion of insulin in pancreatic [beta] cells. Additionally, the concentration of serotonin in the blood of mice fed a high fat diet was much higher than that of lean control mice. These studies suggest that serotonin may play important roles with regard to glucose and lipid metabolism. In this study, we have investigated the physiological effect of peripheral serotonin on the plasma levels of metabolites in both mice and sheep.[br][Results][br]Mice (C57BL/6, male, 7 weeks old) were fasted for 12 h before the experiment. After the intraperitoneal injection of 1 mg serotonin, the plasma glucose, insulin, triglyceride, NEFA, cholesterol and bile acids concentrations were measured. Plasma glucose and insulin significantly elevated after the injection. Plasma triglyceride, cholesterol and NEFA concentrations decreased. The concentration of bile acids in plasma increased after injection. These changes in plasma levels of metabolites and hormone were induced by several kinds of serotonin receptors using experiments of serotonin receptor antagonists. Wethers (crossbred, averaging 47.2 [plusmn] 7.8 kg of body weight) were fasted for 24 h before the experiment, and then intravenous injected with serotonin (40 [mu]g/kg body weight). Serotonin also induced the elevations of plasma glucose and insulin concentrations through the same serotonin receptor in wethers as well as in mice. On the other hand, the concentrations of plasma triglyceride and NEFA increased and the level of plasma bile acids decreased by serotonin injection. Additionally, serotonin did not affect the plasma cholesterol concentration. These results reveal that there are quit differences in the physiological functions of peripheral serotonin against lipid metabolism between mice and sheep.[br][br](1) Watanabe et al., Endocrinology 2010; 151:4776-4786.[br][br]Nothing to Disclose: HW, RS, TN, HT, YT, KS, YN, ST, NO, DWH, NS, HS, KW, HA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 598 55 297 SAT-194 PO14-02 Saturday 236 2012


237 ENDO12L_SAT-195 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Insulin and Leptin Action on Kiss1 Neurons and the Regulation of Puberty and Reproduction Xiaoliang Qiu, Abigail Dowling, Joseph Marino, Latrice Faulkner, Laura Nedorezov, Jennifer Hill University of Toledo, Toledo, OH The neuropeptide kisspeptin, encoded by the Kiss1 gene, is necessary for puberty and fertility. Humans and mice with loss-of-function mutation of Kiss1 or its receptor are infertile due to hypogonadotropic hypogonadism. Insulin and leptin receptors are widely distributed throughout the hypothalamus and play a pivotal role in the regulation of energy balance, metabolism, and reproduction. Recent data show that leptin[apos]s effect on puberty in mice (KiSS1-Cre, ObR[sup]flox/flox[/sup]) does not require KiSS1 neurons. In order to study whether insulin -sensing plays an important role in Kiss1 neurons, we generated Kiss1 neuron specific insulin receptor knockout mice (Kiss1-Cre, IR [sup]flox/flox[/sup] mice). In addition, we produced Kiss1 neuron-specific insulin receptor and leptin receptor knockout mice (Kiss1-Cre, IR [sup]flox/flox[/sup] ObR[sup] flox/flox[/sup] mice) to look at the potential synergic role of the two receptors in the regulation of Kiss1 neurons. Here, we describe effects on vaginal opening, first estrus cycle, and fertility. Kiss1-Cre, IR [sup]flox/flox[/sup] female mice delayed vaginal opening (31.4[plusmn]0.5 VS 34.0[plusmn]0.9 days) and first estrus cycle (39.6[plusmn]1.2 VS 44.1[plusmn]1.6 days). Luteinizing hormone (LH) in Kiss1-Cre, IR [sup]flox/flox[/sup] mice were lower compared with control on early puberty 31 days old, but not adult. Body weight, food intake, and glucose regulation were comparable. Kiss1-Cre, IR [sup]flox/flox[/sup] ObR[sup] flox/flox[/sup] mice were examined for changes in pubertal timing, reproduction and metabolism as well. These results have important implications for understanding how insulin and leptin signaling in Kiss1 neurons may be involved in puberty and reproduction.[br][br]Sources of Research Support: NIH grant R00HD056491 to JWH.[br][br]Nothing to Disclose: XQ, AD, JM, LF, LN, JH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2272 55 298 SAT-195 PO14-02 Saturday 237 2012


238 ENDO12L_SAT-196 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Reduction of Glucose-6-Phosphate Transporter Induces Suppression of 11[beta]-Hydroxysteroid Dehydrogenase Type 1 Expression and Improvement of Glucose Homeostasis in Diabetic Mouse Yanjun Liu, Ying Wang, Yuichi Nakagawa, Wei Wang, Alexei Lyzlov, Kabirullah Lutfy, Theodore C Friedman Charles Drew University of Medicine and Science, Los Angeles, CA; Hamamatsu University School of Medicine, Hamamatsu, Japan Pre-receptor activation of glucocorticoids via 11[beta]-hydroxysteroid dehydrogenase type 1 (11[beta]-HSD1) has been identified as an important mediator of metabolic syndrome. Hexose-6-phosphate dehydrogenase (H6PDH) mediates 11[beta]-HSD1 amplifying tissue glucocorticoid production by driving intracellular NADPH exposure to 11[beta]-HSD1 and requires glucose-6-phosphate transporter (G6PT) to maintain its activity. To test the efficiency of reducing G6PT expression to treat hyperglycemia and insulin resistance, we targeted the G6PT gene in type 2 diabetic mice using G6PT ASO specific to G6PT and explored the impact of G6PT on the pre-receptor metabolism of glucocorticoids in type 2 diabetes and obesity. We observed that G6PT ASO treatment of db/db mice markedly reduced hepatic G6PT mRNA and protein levels and improved glucose homeostasis and insulin resistance. The reduction of hepatic G6PT expression paralleled the suppression of 11[beta]-HSD1 and H6PDH production in the liver. These findings suggest that G6PT plays an important role in the modulation of pre-receptor activation of glucocorticoids and provide new insights into the role of G6PT in the development of type 2 diabetes.[br][br]Sources of Research Support: Y. Liu is supported by NIH grants SC1DK087655.[br][br]Nothing to Disclose: YL, YW, YN, WW, AL, KL, TCF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1060 55 299 SAT-196 PO14-02 Saturday 238 2012


239 ENDO12L_SAT-197 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Glucagon-Induced Phosphorylation of the Salt-Inducible Kinase-2, a Key Mediator That Controls Hormone-Regulated Hepatic Gluconeogenesis Kashyap Patel, Olga Goransson, Calum Sutherland, Kei Sakamoto College of Life Sciences, University of Dundee, Dundee, UK; Lund University, Lund, Sweden; Ninewells Hospital and Medical School, University of Dundee, Dundee, UK The hormones insulin and glucagon are central to regulating glucose homeostasis via controlling hepatic gluconeogesis and peripheral glucose uptake. When the regulation of glucose homeostasis by these hormones dysfunctions, metabolic disorders develop, such as type 2 diabetes. Thus deciphering the details of molecular and cellular signaling network controlled by insulin and glucagon promises to provide new strategies for treating type 2 diabetes. Recent work has proposed that the salt inducible kinase-2 (SIK2), a closely related member of AMP-activated protein kinase (AMPK), plays a key role as a signaling mediator in the control of hormone-regulated hepatic gluconeogenesis. The aim of current study was to identify novel and key phosphorylation sites on SIK2 regulated by glucagon and insulin and elucidate the molecular mechanism by which SIK2 is regulated by these hormones in the liver. We have performed a phospho-peptide mapping analysis by a mass-spectrometry using recombinant SIK2 proteins isolated from mouse primary hepatocytes with or without glucagon or insulin treatment. This revealed that glucagon caused an increase in several phosphorylation sites including Ser358 and Ser587, but not Thr175 site in the T-loop of kinase domain, and these results were validated using phospho-specific antibodies to each site. Interestingly, but contrary to the previous work, we failed to see an increase in Ser358 phosphorylation in response to insulin at multiple time points measured (5-40 min) in mouse primary hepatocytes. To investigate if the phosphorylation of SIK2 by glucagon has impact on SIK2 kinase activity, endogenous SIK2 was immunoprecipitated from glucagon-treated mouse primary hepatocytes and in vitro kinase assay was performed. We observed no significant change in SIK2 activity between non-treated and glucagon-stimulated hepatocytes. We also performed mutagenesis analysis and observed that activity of Ser358Ala or Ser587Ala mutant SIK2 proteins isolated from HEK293 cells was similar compared to that of wild-type SIK2. Taken together, we have identified glucagon-regulated phosphorylation sites on SIK2, which do not appear to be critical for directly controlling SIK2 enzymatic activity, but may be important for localization or protein-protein interaction to regulate target proteins and glucose metabolism in hepatocytes.[br][br]Sources of Research Support: Wellcome Trust Grant awarded to KA.[br][br]Nothing to Disclose: KP, OG, CS, KS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1673 55 300 SAT-197 PO14-02 Saturday 239 2012


240 ENDO12L_SAT-198 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Hyperinsulinemia Promotes Metastasis to the Lung in a Mouse Model of Her2-Mediated Breast Cancer Rosalyn D Ferguson, Dara Cohen, Nyosha Alikhani, Aviva Tobin-Hess, Ruslan Novosyadlyy, Nadine Haddad, Derek Leroith Mount Sinai School of Medicine, New York, NY The Her2 oncogene is expressed in around 30% of human breast cancers and is associated with frequent metastatic progression and poor outcome.Epidemiological studies show that breast cancer incidence and mortality rates are higher in women with type 2 diabetes. Here we use a mouse model of Her2-mediated breast cancer on a background of hyperinsulinemia to determine how elevated circulating insulin levels affect Her2-mediated primary tumor growth and subsequent progression to lung metastasis. Hyperinsulinemic (MKR) mice were crossed with doxycycline-inducible Neu (rtTANeu) mice to produce the rtTANeuMKR mouse model. Both rtTANeu controls and rtTANeuMKR mice were administered doxycycline in drinking water to induce mammary tumor formation. At time of sacrifice total mammary tumor weight from both groups was measured and lungs were removed so that metastases could be quantified. After 90 days on doxycycline, although no significant increase in tumor weight was observed in rtTANeuMKR compared to rtTANeu mice, the number of lung metastases was significantly higher in rtTANeuMKR compared to rtTANeu mice (rtTANeuMKR 16.41 [plusmn] 4.18 vs. rtTANeu 5.36 [plusmn] 2.72). We analyzed signaling pathways downstream of the insulin receptor (IR)/insulin-like growth factor receptor 1 (IGF-1R) in tumor tissues. As well as finding increased activation of S6 kinase, an important mediator of cell proliferation, we also observed an increase in Snail, a transcription factor which is known to be important for the epithelial to mesenchymal transition (EMT) in breast cancer metastasis. We conclude that while primary tumors were similar in size, the hyperinsulinemia in the rtTANeuMKR mice resulted in increased EMT changes and therefore more aggressive primary tumors and a higher numbers of metastatic events.[br][br]Nothing to Disclose: RDF, DC, NA, AT-H, RN, NH, DL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 537 55 301 SAT-198 PO14-02 Saturday 240 2012


241 ENDO12L_SAT-199 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Hypoglycemic Effect of Aqueous Extract of [italic]Carica papaya[/italic] Leaf in Streptzotocin-Induced Diabetic Rats Isela Juarez Rojop, P H Miranda-Osorio, Teresa Ramon-Frias, Hidemi Aguilar-Mariscal, Jorge Ble-Castillo, A Rodriguez-Hernandez, Maria A Jimenez-Santos, Juan C Diaz-Zagoya Universidad Ju[agrave]rez Aut[oacute]noma de Tabasco, Villahermosa, Mexico; Universidad Ju[agrave]rez Aut[oacute]noma de Tabasco, Comalcalco, Mexico Diabetes is one of the major global public health problems and rapidly getting worse particularly in the developing nations (Unwin et al., 2009). In addition, the use of plants herbal remedies against diseases that constitutes economic problems such as diabetes (Adisa et al., 2011). [italic]Carica papaya[/italic] belongs to the family of [italic]Caricaceae[/italic] and several species of [italic]Caricaceae[/italic] have been used as remedy against a variety of diseases (Perez et al., 2003). The present study was to determine the effect of aqueous extract of [italic]C. papaya[/italic] leaf in streptozotocin (STZ)-induced diabetes in rats. Diabetes was induced in male Wistar rats (250-300g) using a single intraperitoneal injection of STZ (60mg/Kg). An experimental groups of non diabetic (ND) and diabetic (D) rats received an aqueous extract of [italic]C. papaya[/italic] a doses (3.5, 7.5 and 15 g/500 ml) for 30 days while other group ND and D received water for the same length of time. At the end of treatment, the animals were sacrificed by decapitation and blood sample was collected. Level of serum glucose, cholesterol, triglyceride and HDL-cholesterol concentrations were determined by using spectrometric methods. Level of insulin was assayed by an ELISA kit. Treatment with extract of [italic]C. papaya[/italic] reduced a dose-dependent manner level of glucose in D rats. Plasma insulin levels were increased by treatment in ND but not modify in D rats. Additionally,[italic] C. papaya[/italic] diminished the triglyceride level in D rats. The results indicated that aqueous extract of [italic]C. papaya[/italic] showed a clear hypoglycaemic effect in diabetic rats. Such an effect cannot be mediated by an enhanced insulin secretion.[br][br]Sources of Research Support: PFICA.UJAT-2010-C06-10.[br][br]Nothing to Disclose: IJR, PHM-O, TR-F, HA-M, JB-C, AR-H, MAJ-S, JCD-Z 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1064 55 302 SAT-199 PO14-02 Saturday 241 2012


242 ENDO12L_SAT-200 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Characterization of Amylin Receptor and Amylin-Regulated Insulin Secretory Pathways in Rat Insulinoma and Human Pancreatic Islet Cells Yasemin Cagil, Saurabh Trikha, Aleksandar Jeremic The George Washington University, Washington, DC Human amylin is a 37-amino acid peptide hormone that is produced and co-secreted with insulin by pancreatic islet beta-cells. In contrast to insulin, the physiological role of amylin is less clear. Studies indicate that amylin may, together with insulin, regulate secretion of insulin and other hormones from islets through autocrine/paracrine signaling mechanisms. However, the exact mechanism and receptors involved in regulatory effect of amylin on insulin secretion needs to be established. Hence in this study, the modulatory effect of amylin on basal and glucose-evoked insulin release in cultured rat insulinoma (RINm5F) cells and human islet cells were investigated using ELISA immunoassay. Low concentrations of amylin (1-10 nM) inhibited both basal and glucose-evoked insulin secretion in rat and human islets. This inhibitory effect of amylin on insulin secretion was also observed in cultures containing high (20 mM) potassium chloride. This result suggests that amylin acts downstream or at the level of glucose-evoked depolarization of beta-cells. This finding is consistent with previous studies showing the hyper-polarizing effect of human amylin on electrical activity of islet beta-cells. The physiological range (10[sup]-10 [/sup]- 10[sup]-8 [/sup]M) in which amylin modulates insulin release suggests involvement of amylin receptor in this secretory process. To test this hypothesis, we examined expression of amylin receptor (AM-R) constituents, calcitonin receptor (CT-R) and receptor activity modifying protein (RAMP), in two cell types by western blot and immunocytochemistry. Western blot analysis demonstrated expressions of type-1 and type-2 AM-R isoforms in human and rat pancreatic cells, respectively. Immunoconfocal microscopy further showed increased co-localization of CT-R and RAMP-2 on plasma membrane of RINm5F cells upon exposure to low doses of human amylin (1-10 nM) for 24 hours, indicating cells[apos] capacity for amylin sensing, which may play a role in regulation of insulin secretion. Further studies are needed to clarify causal connection between amylin receptor expression and amylin biological function including regulation of insulin release in pancreatic islets.[br][br]Nothing to Disclose: YC, ST, AJ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1665 55 303 SAT-200 PO14-02 Saturday 242 2012


243 ENDO12L_SAT-201 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Uncoupling Protein 3 Inhibits Glucose Oxidation in Muscle Cells Romain Harmancey, Truong Nguyen Lam, Heinrich Taegtmeyer University of Texas Medical School at Houston, Houston, TX Glucose and fatty acids are competing substrates for energy provision in muscle. Fatty acid-mediated inhibition of glucose oxidation (Randle cycle) results from the inhibition of phosphofructokinase and pyruvate dehydrogenase (PDH). However, additional control sites may exist. Because fatty acids are potent activators of uncoupling protein (UCP) 3 expression, we proposed that the mitochondrial protein contributes to the inhibition of glucose oxidation in myocytes. L6 rat myoblasts were differentiated into myocytes, and changes in UCP3 expression levels were correlated to PDH activity by determination of its inhibitory phosphorylation status on Ser232, Ser293, and Ser300. Cellular rates of glucose oxidation were determined by quantitative collection of [[sup]14[/sup]C]O[sub]2 [/sub]following incubation with [U-[sup]14[/sup]C]glucose. Glycolytic flux was determined indirectly by quantification of lactate released in the incubation medium. To further examine the effect of UCP3 on glucose metabolism, we then transfected L6 myocytes with two siRNAs targeting the mitochondrial protein. UCP3 was not expressed in proliferating L6 myoblasts, but there was a 16-fold increase in UCP3 expression, both at the mRNA and protein levels, seven to eight days after the initiation of myogenic differentiation. The rise in UCP3 expression was tightly correlated to a gradual increase in PDH phosphorylation on Ser232 (r=0.9753; [italic]p[/italic][lt]0.001), Ser293 (r=0.9657; [italic]p[/italic][lt]0.01), and Ser300 (r=0.9461; [italic]p[/italic][lt]0.01). Compared to myoblasts, the rates of glucose oxidation were consequently reduced by more than 50% in the differentiated myocytes. In the meantime, lactate release was increased, suggesting that glycolysis becomes gradually uncoupled from glucose oxidation. The knockdown of the UCP3 transcript approached 70% with both siRNA sequences, resulting in a more than 60% decrease of UCP3 protein levels. This reduction in the protein level of UCP3 was sufficient to increase the rates of glucose oxidation in myocytes. We conclude that reduced UCP3 expression favors glucose oxidation and improves the coupling of glucose metabolism in cultured myocytes. Fatty acid-mediated expression of UCP3 may represent an additional control point for fatty acid inhibition of glucose oxidation.[br][br]Sources of Research Support: NHLBI Grant 5R01HL061483; Postdoctoral fellowship 09POST2060155 from the American Heart Association awarded to RH.[br][br]Nothing to Disclose: RH, TNL, HT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1650 55 304 SAT-201 PO14-02 Saturday 243 2012


244 ENDO12L_SAT-202 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Testosterone Deficiency Is Associated with Decreased Expression of Muscle GLUT4 and Hexokinase 2 as Key Regulators of Insulin Sensitivity in the Testicular Feminized Mouse David S McLaren, Daniel M Kelly, Samia Akhtar, Kevin S Channer, Thomas H Jones University of Sheffield, Sheffield, UK; Royal Hallamshire Hospital, Sheffield, UK; Barnsley Hospital NHS Foundation Trust, Barnsley, UK Testosterone deficiency is common in men with type 2 diabetes (T2D). We have shown testosterone replacement therapy (TRT) improves insulin resistance and glycaemic control. The mechanisms by which testosterone mediates this action are unknown but may be due to a combination of effects on muscle, liver and adipose tissues. This study investigates the expression of Glut4 and HK2, (two key proteins involved in insulin sensitivity) in muscle tissue of the testicular feminised (Tfm) mouse which exhibit non-functional androgen receptors and low circulating levels of testosterone.[br]Tfm mice were fed a high-cholesterol diet ad libitum for 28 weeks and received either physiological testosterone replacement (mixed testosterone esters, Sustanon100) or placebo (saline) and were compared to wild-type littermates (WT). Striated muscles tissue was collected from the thigh. Expression of Glut 4 and HK2 mRNA and protein expression was analysed by qPCR and Western blotting.[br]There was a significant decrease in the relative mRNA expression of Glut-4 (0.59[plusmn]0.14, p=0.02) and HK2 (0.5[plusmn]0.16,p=0.01) in Tfm mice compared to WT. TRT did not significantly alter mRNA expression of Glut4 or HK2. Western blotting confirmed reduced protein expression of HK2 (0.33[plusmn]0.09vs0.04[plusmn]0.01,p=0.005) and Glut4 (1.05[plusmn]0.22vs0.57[plusmn]0.07,p=0.037) in Tfm mice compared to WT. TRT showed no change in HK2 protein expression compared to Tfm placebo (0.05[plusmn]0.02vs0.04[plusmn]0.01,p=0.627), but an increase in Glut4 was observed (1.09[plusmn]0.16vs0.57[plusmn]0.07,p=0.005).[br]Testosterone deficiency is associated with decreased expression of Glut4 and HK2. TRT did not have any effect on HK2 in the Tfm mouse suggesting that testosterone acts via the androgen receptor to stimulate HK2 expression. Although testosterone failed to increase Glut4 mRNA in the Tfm an increase in protein was observed indicating that post-translational mechanisms are involved. This study shows that testosterone has important actions on modulating glucose metabolism in muscle. This may explain part of the mechanism by which testosterone improves insulin sensitivity in T2D.[br][br]Nothing to Disclose: DSM, DMK, SA, KSC, THJ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1453 55 305 SAT-202 PO14-02 Saturday 244 2012


245 ENDO12L_SAT-203 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Potential Mechanism of Type 2 CRF Receptor-Mediated Skeletal Muscle Insulin Resistance Hongxia Chao, Haochen Li, Peilin Chen, Chien Li University of Virginia Health System, Charlottesville, VA Insulin resistance for glucose disposal by skeletal muscle (SM) is an essential and perhaps primary defect for Type 2 diabetes and metabolic syndrome. The underlying mechanism of muscle insulin resistance has not been fully delineated and is likely due to an interaction between multiple extrinsic and intrinsic factors. Urocortin 2 (Ucn 2), a member of the corticotropin-releasing factor (CRF) family, and its selective receptor, the type 2 CRF receptor (CRFR2), is highly expressed in SM. It has been suggested that CRFR2 and its ligand may be involved in regulating glucose homeostasis in muscle.[br]In the present study, we found that CRFR2 expression was greatly increased in the muscle of obese diabetic Ob/Ob mice, but was significantly reduced in the muscle of exercised mice compared to that of sedentary control mice. Consistent with our in vivo observation, CRFR2 expression in insulin resistant C2C12 myotubes induced by either chronic insulin or palmitic acid treatment was greatly elevated compared to that of control cells. Functional study showed overexpression of CRFR2 in SM cells blocked insulin-induced glucose uptake. Conversely, reduced CRFR2 expression improved insulin sensitivity in insulin resistant myotubes. Moreover, we found that cAMP is required for the inhibitory effect of Ucn 2 on insulin-induced glucose uptake and blocking Epac and protein kinase A, two prominent cAMP downstream effectors, failed to modulate the effect of Ucn 2. Interestingly, pretreating cells with rapamycin prevented the inhibitory effect of Ucn 2 on insulin-induced glucose uptake. Furthermore, Ucn 2 attenuated insulin-induced tyrosine608 phosphorylation of insulin receptor substrate 1 (IRS1) and increased phosphorylation of IRS1 serine 636/639 residues, whereas treatment with rapamycin reversed the phosphorylation of these residues. Finally, Ucn 2 failed to inhibit glucose uptake induced by platelet-derived growth factor, which stimulates PI3-kinase signaling in an IRS1 independent manner. This result further supports the notion that CRFR2 signaling modulates insulin[apos]s action at the level of IRS1.[br]In summary, we show CRFR2 expression is closely correlated with the status of muscle insulin sensitivity and CRFR2 signaling plays a critical role in modulating insulin-induced glucose uptake. Furthermore, we found that IRS1 is the key nodal point where CRFR2 signaling modulates insulin action and the effect of CRFR2 may be mediated by a novel cAMP-mTOR signaling pathway.[br][br]Sources of Research Support: NIH Grant DK078049 awarded to CL.[br][br]Nothing to Disclose: HC, HL, PC, CL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2026 55 306 SAT-203 PO14-02 Saturday 245 2012


246 ENDO12L_SAT-204 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Adiponectin Restores Insulin Responsiveness in Human Neuroblastoma Cell Line SH-SY5Y Emna Chaabouni, Gael Lemerle, Yacir Benomar, Michel Parquet, Mohammed Taouis University Paris-Sud, Orsay, France; CNRS, Orsay, France Adiponectin regulates energy homeostasis through the modulation of glucose and fatty acid metabolism in peripheral tissues. However, its central effect on energy balance remains unclear and controversial. Recent advances in our understanding of the signal transduction mechanisms used by adiponectin in the periphery and in the hypothalamus suggest that intracellular cross-talk between adiponectin, leptin and insulin may occur. In the present study we aimed to study the impact of adiponectin on insulin responsiveness in human neuroblastoma cell line SH-SY5Y. In order to study the potential cross talk between adiponectin and insulin signaling pathways, it is necessary to demonstrate that SH-SY5Y cells express insulin and adiponectin receptors. We have previously shown that these cells express insulin receptor, and here using immunohistochemical approach we show that SH-SY5Y cells express both AdipoR-1 and Adipo-R2. Then we studied the functionality of these receptors by measuring the phosphorylation of AMPkinase in response to adiponectin. We show that adiponectin increased AMP-K phosphorylation in a dose dependent manner. Since in literature adiponectin is considered as an anti-diabetic hormone, we studied the plausible impact of adiponectin in delaying or abolishing insulin resistance associated to an overexposure of cells to insulin. Firstly, we show that a pretreatment with insulin significantly down-regulated insulin receptor expression at the protein level which is accompanied by an impairment of insulin-dependent Akt phosphorylation. To investigate the potential anti-insulin resistance effect of adiponectin, we studied the impact of adiponectin on Akt phosphorylation after insulin overexposure. As expected, in the absence of insulin pretreatment, acute insulin stimulation increased significantly Akt phosphorylation. This effect was completely abolished when cells were overexposed to insulin. Interestingly, when cells were overexposed to both insulin and adiponectin the insulin-dependent Akt phosphorylation was completely restablished protecting then cells from insulin resistance. In addition, we show that adiponectin exerted, most likely, its effect by reducing the insulin-dependent down-regulation of insulin receptors.[br]In conclusion, we show that adiponectin in a neuronal cells is able to maintain insulin responsiveness in an environment favoring insulin resistance through probably the inhibition of insulin receptor down-regulation.[br][br]Nothing to Disclose: EC, GL, YB, MP, MT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1182 55 307 SAT-204 PO14-02 Saturday 246 2012


247 ENDO12L_SAT-205 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Low-Carb/High-Fat Diets Reduce Fasting Glucose and Insulin Levels, but Are Associated with Insulin Resistance and Reduced Pancreatic [beta]-Cell Mass in Rats Maximilian Bielohuby, Stephanie Sisley, Nadja Herbach, Darleen Sandoval, Juliane Ramisch, Joyce Sorrell, Dominik Menhofer, Rudiger Wanke, Kerstin Stemmer, Matthias H Tschop, Randy J Seeley, Martin Bidlingmaier Medizinische Klinik und Poliklinik IV, Klinikum der LMU, Munich, Germany; University of Cincinnati, Cincinnati, OH; College of Medicine, University of Cincinnati, Cincinnati, OH; LMU, Munich, Germany; Helmholtz Centre for Health and Environment and Technical University, Munich, Germany Low-carbohydrate/high fat (LCHF) diets are claimed to induce weight loss in overweight subjects and to ameliorate glucose metabolism. To investigate the effects of LCHF diets on glucose and insulin metabolism independent of the energetic intake, we pair-fed male rats isoenergetic amounts of standard rodent chow (CH) or one of two different LCHF diets (% of metabolizable energy, fat/protein/CHO: LCHF-1 (78.7/19.1/2.2), LCHF-2 (92.8/5.5/1.7) and CH (16.7/19./64.3)). After 3 weeks of feeding the respective diets, oral glucose tolerance tests (OGTT), insulin tolerance tests (ITT) and hyperinsulinemic-euglycemic clamps were performed (n=4-5/challenge test and diet group). After 4 weeks on the diets, rats were fasted for 6h and sacrificed for collection of trunk blood and pancreas. Serial pancreas sections from rats which have not been previously challenged were used to determine total [beta]-cell volume by quantitative stereological techniques (n=5/diet group). Results: Basal fasting glucose and insulin levels were lower in rats fed LCHF vs. CH diets (p[lt]0.05). Total pancreas volumes in rats fed LCHF diets were 20% (LCHF-1; p[lt]0.01) and 30% (LCHF-2; p[lt]0.001) lower compared to CH. Quantitative stereological analyses revealed that both LCHF diets led to significantly (p[lt]0.001) lower total [beta]-cell volumes, both absolute and relative to pancreas volume and body weight. During OGTT, the area under the glucose curve was significantly higher in rats fed both LCHF diets, but insulin secretion was not affected. The decline in glucose during ITT was significantly delayed in LCHF vs. CH fed animals. Clamp studies showed that the glucose infusion rate was about 3-fold higher (p[lt]0.05) and endogenous glucose production (EGP) over than 50% lower in rats fed CH. In contrast, EGP was not inhibited and clearance of glucose was significantly reduced by both LCHF diets. In summary, feeding LCHF diets led to lower fasting glucose and insulin levels. At first sight these data support a beneficial role of LCHF diets for glucose metabolism. However, dynamic challenge tests revealed that rats fed LCHF diets developed insulin resistance. Moreover, lower basal insulin levels of rats fed LCHF diets are most likely explained by a loss of pancreatic [beta]-cell mass. This occurred independent of the relative abundance of fat and protein in LCHF diets and in the absence of increased energetic intake. Our data clearly argue against a beneficial effect of LCHF diets on glucose and insulin metabolism.[br][br]Disclosures: DS: Research Funding, Johnson & Johnson, Novo Nordisk, Pfizer, Inc.; Speaker, Novo Nordisk; MB: Study Investigator, Chiasma. Nothing to Disclose: MB, SS, NH, JR, JS, DM, RW, KS, MHT, RJS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 951 55 308 SAT-205 PO14-02 Saturday 247 2012


248 ENDO12L_SAT-206 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) A New Syntaxin4 Binding Partner Tctex1d2 Is Specifically Expressed in Adipocytes and Inhibits GLUT4 Translocation through Interference of Doc2b-Syntaxin4 Interaction Atsushi Nohara, Shuichi Okada, Tsugumichi Saito, Kihachi Ohshima, Jeffrey E Pessin, Masatomo Mori Gunma University Graduate School of Medicine, Maebashi, Japan; Gunma University, Maebashi, Japan; Albert Einstein College of Medicine, New York, NY ctex1d2 is a novel gene that encodes for a functionally unknown protein that contains a Tctex1 domain found in dynein light chain family members. Examination of gene expression during adipogenesis demonstrated a marked increased in Tctex1d2 gene expression that was essentially undetectable in pre-adipocytes and markedly induced during 3T3-L1 adipocyte differentiation. Acute insulin stimulation (up to 30 min) in 3T3-L1 adipocytes over expressing Tctex1d2 had no effect on the phosphorylation of the insulin receptor [beta] subunit, IRS-1, Erk, or Akt. However Tctex1d2 overexpression significantly inhibited insulin-stimulated GLUT4 translocation. Syntaxin4 is a binding partner for Doc2b and the v-SNARE protein VAMP2 that are required for insulin-stimulated GLUT4 translocation. Over expression of Tctex1d2 was found to associate with Syntaxin4 and prevent the insulin-stimulated association of Doc2b. Based upon these data, we hypothesize that Tctex1d2 and Doc2b share binding sites on Syntaxin4 thereby inhibiting insulin induced GLUT4 translocation through interference of Doc2b-Syntaxin4 interaction.[br][br]Nothing to Disclose: AN, SO, TS, KO, JEP, MM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 32 55 309 SAT-206 PO14-02 Saturday 248 2012


249 ENDO12L_SAT-207 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Estrogen Facilitates Insulin Signaling in a Leptin-Dependent Manner Shiva PD Senthil Kumar, Xian Liu, Haifei Shi Miami University, Oxford, OH Estrogens regulate glucose homeostasis. Postmenopausal women with decreased levels of endogenous estrogen have increased risk of insulin resistance, and treatment with estradiol (E2; the primary form of estrogen) improves insulin sensitivity. Leptin and insulin, two hormones that regulate energy homeostasis, have overlapping intracellular PI3/Akt signaling pathway and elicit similar metabolic actions. We hypothesize that the insulin sensitizing action of estrogen on insulin-targeted tissues is influenced by leptin. Three male mouse models were used in this study, standard chow-fed lean mice with low circulating leptin level and normal leptin sensitivity, high-fat diet-induced obese mice with high circulating leptin level and leptin resistance, and leptin-deficient [italic]ob/ob[/italic] mice. First the role of E2 in insulin sensitivity was assessed using intraperitoneal insulin tolerance test in combination with injections of E2 or its vehicle. Injection of a sub-threshold dose of insulin (0.5 U/kg) did not significantly change glucose levels of lean mice; however, when the lean mice were co-injected with 200 mg/kg E2 and 0.5U/kg insulin, their glucose levels declined more rapidly, with significantly greater glucose disappearance rate and lower glucose levels, than the lean mice injected with 0.5U/kg insulin and vehicle for E2. The obese mice co-injected with 1U/kg insulin and E2 cleared glucose efficiently, with their glucose disappearance rates being faster than the obese mice injected with insulin alone. In contrast, [italic]ob/ob[/italic] mice co-injected with E2 and insulin had similar glucose levels as the [italic]ob/ob[/italic] mice treated with insulin alone. Thus, administrations of insulin and E2 had greater effects in lowering glucose levels than insulin alone in wildtype, but not in leptin deficient mice. Second E2[apos]s effects on insulin and leptin signaling pathways were investigated. Phosphorylation of Akt (common markers for insulin and leptin signaling), MAPK (marker for insulin signaling), and STAT3 (marker for leptin signaling) in metabolic tissues was measured by Western blot analysis. The ratios of pAkt/total Akt, pMAPK/total MAPK, and pSTAT3/total STAT3 in skeletal muscle, white adipose tissue and the hypothalamus of wildtype mice increased after co-injection of E2 and insulin comparing to insulin injection alone. Our data indicate that acute E2 improves insulin sensitivity and suggest that this effect is related to an activated insulin and leptin signaling in the periphery and the CNS.[br][br]Sources of Research Support: National Institutes of Health National Institute of Diabetes and Digestive and Kidney Diseases [Grant 1R15DK090823-01 to HS] and American Heart Association [Grant 10SDG4520028 to HS].[br][br]Nothing to Disclose: SPDSK, XL, HS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1233 55 310 SAT-207 PO14-02 Saturday 249 2012


250 ENDO12L_SAT-208 POSTER SESSION: Insulin [amp] Nutrient Metabolic Action in Animal Models (1:30 PM-3:30 PM) Adult Rats Exposed to Ethanol In Utero Have Increased Hepatic Gluconeogenesis Due to Oxidative Stress-Induced Histone Deacetylases Xing-Hai Yao, Hoa Khanh Nguyen, Jonghan Lee, Gregoire Nyomba University of Manitoba, Winnipeg, Canada [bold]Introduction:[/bold] Prenatal ethanol (EtOH) increases hepatic gluconeogenesis in adult rat offspring by unknown mechanism[bold]. [/bold][br][bold]Methods:[/bold] Four month-old male rat offspring exposed to EtOH with or without tauroursodeoxycholic acid (TUDCA) during pregnancy days 1-7 (early), 8-14 (mid) and 15-21 (late) were compared with those from pair-fed (PF) dams. We performed pyruvate challenge test and measured liver reactive oxygen species (ROS), phosphoenolpyruvate carboxykinase (PEPCK), glucose 6-phosphatase (G6Pase), nuclear histone deacetylases (HDACs), and acetylated Foxo1 (AcFoxo).[br][bold]Results:[/bold] EtOH vs PF vs TUDCA (Mean[plusmn]SE, *P[lt]0.05, **P[lt]0.01):[br]-pyruvate induced glycemia (AUC): early: 1485[plusmn]93 vs 1159[plusmn]133* vs 1188[plusmn]76*; mid: 1565[plusmn]55 vs 1229[plusmn]67** vs 1259[plusmn]83*; late: 1734[plusmn]70 vs 1242[plusmn]84** vs 1375[plusmn]115*;[br]-ROS: early: 118.84[plusmn]13.07 vs 72.61[plusmn]6.65** vs 81.83[plusmn]9.56*; mid: 124.25[plusmn]12.51 vs 65.46[plusmn]9.02** vs 79.91[plusmn]8.58*; late: 119.68[plusmn]14.71 vs 73.52[plusmn]9.49* vs 79.62[plusmn]7.90*;[br]-PEPCK: early: 1.60[plusmn]0.15 vs 1.07[plusmn]0.13* vs 1.30[plusmn]0.13; mid: 2.03[plusmn]0.21 vs 1.14[plusmn]0.10** vs 1.20[plusmn]0.15*; late: 1.90[plusmn]0.29 vs 0.91[plusmn]0.12** vs 1.06[plusmn]0.09**;[br]-G6Pase: early: 1.63[plusmn]0.24 vs 0.95[plusmn]0.13* vs 1.33[plusmn]0.13; mid: 1.47[plusmn]0.12 vs 0.97[plusmn]0.12* vs 1.21[plusmn]0.12; late: 1.42[plusmn]0.08 vs 1.05[plusmn]0.10* vs 1.25[plusmn]0.12;[br]-HDAC activity: early: 0.92[plusmn]0.07 vs 0.43[plusmn]0.05** vs 0.63[plusmn]0.09*; mid: 0.75[plusmn]0.05 vs 0.45[plusmn]0.05** vs 0.52[plusmn]0.05*; late: 0.72[plusmn]0.07 vs 0.38[plusmn]0.05** vs 0.52[plusmn]0.05*;[br]-HDAC3: early: 1.73[plusmn]0.12 vs 0.94[plusmn]0.06** vs 1.07[plusmn]0.11**; mid: 1.60[plusmn]0.13 vs 1.05[plusmn]0.08** vs 1.16[plusmn]0.09*; late: 2.73[plusmn]0.25 vs.98[plusmn]0.13** vs 1.25[plusmn]0.09**;[br]-HDAC4: early: 2.04[plusmn]0.27 vs 0.76[plusmn]0.14** vs 1.21[plusmn]0.27*; mid: 1.87[plusmn]0.27 vs 0.96[plusmn]0.10** vs 1.26[plusmn]0.09*; late: 2.37[plusmn]0.47 vs 0.93[plusmn]0.11**vs 1.31[plusmn]0.45*;[br]-HDAC5: early: 1.95[plusmn]0.22 vs 1.09[plusmn]0.12** vs 1.04[plusmn]0.14**; mid: 1.85[plusmn]0.21 vs 1.09[plusmn]0.09* vs 1.17[plusmn]0.21*; late: 2.63[plusmn]0.55 vs 0.72[plusmn]0.17** vs 1.34[plusmn]0.20*;[br]-HDAC7: early: 3.32[plusmn]0.79 vs 0.76[plusmn]0.11** vs 1.47[plusmn]0.33*; mid: 2.56[plusmn]0.55 vs 0.87[plusmn]0.27** vs 1.33[plusmn]0.23*; late: 2.18[plusmn]0.52 vs 0.96[plusmn]0.13* vs 1.05[plusmn]0.14*;[br]-AcFoxo: early: 0.17[plusmn]0.03 vs 0.98[plusmn]0.07** vs 0.46[plusmn]0.05*; mid: 0.22[plusmn]0.06 vs 1.01[plusmn]0.06** vs 0.43[plusmn]0.04*; late: 0.16[plusmn]0.02 vs 1.07[plusmn]0.15** vs 0.48[plusmn]0.05*.[br][bold]Conclusion:[/bold] Adult rats exposed to EtOH in utero, have increased gluconeogenesis, oxidative stress, and HDAC expression, but reduced Foxo acetylation. Oxidative stress likely increases HDACs, which deacetylate Foxo, leading to gluconeogenic genes transcription. These anomalies occur even in early pregnancy and can be prevented by TUDCA administration to the EtOH dams.[br][br]Sources of Research Support: Canadian Diabetes Association grant OG-3-11-3419-BN.[br][br]Nothing to Disclose: X-HY, HKN, JL, GN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 124 55 311 SAT-208 PO14-02 Saturday 250 2012


251 ENDO12L_SAT-209 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Hallucinations Associated with Miglitol Use in a Patient with Chronic Kidney Disease and Hypothyroidism Thet Hlaing, Shilpa Chaudhari, Alan Sacerdote, Gul Bahtiyar Woodhull Medical and Mental Center, Brooklyn, NY; State University of New York (SUNY) Downstate Medical Center, Brooklyn, NY; New York University School of Medicine, New York, NY; St George[apos]s University School of Medicine, Grenada, Grenada A 71 year old woman with Type 2 diabetes mellitus (Type 2DM), chronic kidney disease stage IV (CKD stage IV), primary hypothyroidism, and osteoarthritis whose prescribed treatment included miglitol 50 mg thrice daily with the first bite of meals reported that she suffered visual hallucinations while taking miglitol, which resolved within a few days of stopping the drug. When she resumed miglitol hallucinations recurred within a few days and again resolved within a few days of stopping the drug. At no point were hallucinations associated with any symptom of hypoglycemia or a low finger stick glucose reading. Glycemic control was suboptimal with HbA1c by HPLC = 11.2%. She was still hypothyroid on thyroxine replacement with TSH by chemiluminescence = 39.7 mIU/ml.[br]Although, in general, miglitol is minimally absorbed following oral administration (2-3%) it is mostly eliminated unchanged via the renal route and thus, may accumulate, in the setting of renal failure. Likewise, incompletely compensated hypothyroidism may result in drug accumulation. Once absorbed, miglitol is able to cross the blood/brain barrier and potentially inhibit the alpha-glucosidase enzymes of the the central nervous system (CNS), eg isomaltase, resulting in CNS glycogen accumulation mimicking genetic glycogen storage diseases, eg Pompe disease, which may also present with hallucinations. Inhibition of CNS alpha-glucosidases may also result in formation of aneurysms. Although one series has reported a 2% incidence of hallucinations in patients taking the related drug, acarbose, the risk is not widely appreciated and it is not a reported adverse effect in the package insert for either medication.[br]We conclude that alpha-glucosidase inhibitors should be used with caution in patients with coexisting renal insufficiency and incompletely treated hypothyroidism. CNS changes, in the absence of hypoglycemia should dictate drug discontinuation.[br][br]Ritesh Patil, MD, MPH and Jack L. DePriest, MD, FCCP, FACP. J Gen Intern Med. 2009 May; 24(5): 683[ndash]686.[br][br]Nothing to Disclose: TH, SC, AS, GB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 136 56 312 SAT-209 PO47-02 Saturday 251 2012


252 ENDO12L_SAT-210 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Severe Hyperglycemic Hyperosmolar State and Acute Mental Status Changes in a Child with New-Onset Type 1 Diabetes Mellitus Nohemie Brevil, Priya Khanna Mount Sinai Hospital, Chicago, IL [italic][bold]Introduction:[/bold][/italic] Hyperglycemic hyperosmolar state (HHS) is a life-threatening diabetic emergency that is thought to occur most commonly in adults patients with type 2 diabetes mellitus. However HHS is increasingly being seen in pediatric patients presenting with new onset type 1 and type 2 diabtes mellitus, and is associated with a high mortality rate.[br][italic][bold]Clinical Case:[/bold][/italic] An 8.9-year old, previously healthy, African American female with a low BMI (14.3 kg/m2) presented to our emergency department with history of weakness and altered mental status for two days. After initial evaluation, the patient was admitted to our pediatric intensive care unit (PICU) for mild diabetic ketoacidosis (DKA), hypernatremia, severe dehydration, renal insufficiency, and hyperosmolarity. Her initial lab findings were: Serum glucose 1258 mg/dL (65-99), serum sodium 155 mEq/L (138-145), corrected sodium 173 mEq/L, serum osmolality 412 mOsm/kg (278-305), serum bicarbonate 11 mmol/L (22-29), blood urea nitrogen 90 mg/dL (5-18), creatinine 4.69 mg/dL (0.3-0.7), venous pH 7.11, beta-hydroxybutyrate 15.3 mmol/L (0.1-0.3), urine ketone 10mg/dL. Her hemoglobin A1c was found to be 10.1%. Islet cell antibodies were negative while those to glutamic acid decarboxylase (GAD) were positive at more than 30 U/ml. These results are consistent with HHS in a new onset type 1 diabetic. The patient[apos]s acidosis corrected within six hours of presentation. She continued to have altered mental status and decreased cognition for three days. Her hyperosmolarity and hypernatremia resolved slowly over four days with careful IV hydration. Once her neurologic status returned to baseline and she was ready for oral feeds, she was started on subcutaneous insulin therapy. She was discharged home after one week of hospitalization.[br][italic][bold]Conclusion:[/bold][/italic] This case highlights the fact that pediatric patients with new onset type 1 diabetes mellitus can present with both DKA and HHS. Management strategies are undefined, but we do know that these patients require careful IV hydration and appropriate fluid replacement to improve their clinical outcome and decrease their mortality rate.[br][br]Nothing to Disclose: NB, PK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 957 56 313 SAT-210 PO47-02 Saturday 252 2012


253 ENDO12L_SAT-211 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) A Case of Steroid Diabetes with Severe Acute Pancreatitis after Liraglutide and Bezafibrate Administration Naoki Hiroi, Mariko Sue, Kenzaburo Oda, Yoshie Oka, Rika Shigemitsu, Ryo Iga, Aya Yoshihara, Shuki Usui, Koji Kuboki, Gen Yoshino Toho University, Tokyo, Japan [Introduction] It is reported that incidence of drug induced acute pancreatitis is only 0.2[ndash]2%, however the true incidence is not clear. The mechanism of drug induced damage of the pancreas is not still completely understood.[br][Case] A 58-year-old, nonalcoholic Japanese male with steroid induced diabetes, who had taken prednisolone 30 mg daily for 3 years for rheumatoid arthritis and interstitial pneumonia, presented with a 3 days history of mid-epigastric pain 3 months after treatment of liraglutide 0.6 mg daily and 1 month after treatment of bezafibrate 200mg daily. Levels of fasting plasma glucose and glycosylated hemoglobin were 172 mg/dL and 6.5% (NGSP), respectively. Liver function test results and serum creatinine concentration were almost normal. Level of triglyceride was 155 mg/dL. Serum lipase concentration was 79 U/L (reference range, 11-53 U/L), and serum amylase concentration was 513 U/L (reference range, 24-137 U/L) at admission. Abdominal computed tomography (CT) showed changes consistent with pancreatitis, and the intestinal thickness and ascites. Liraglutide and bezafibrate were discontinued. Conservative therapy for acute pancreatitis resulted in rapid resolution of symptoms, normal lipase concentration (37 U/L).[br][Discussion] In this case, diabetes and hypertriglyceridemia were stable, and the past history which caused pancreatitis was not seen. He took many drugs for treatment of rheumatoid arthritis and interstitial pneumonia, however, administration of liraglutide and/or bezafibrate might cause pancreatitis, because there were no drugs introduced other than liraglutide and bezafibrate newly.[br][Conclusions] Liraglutide and/or bezafibrate should be used cautiously in patients with diabetic patients.This case suggests that liraglutide- and/or bezafibrate-induces pancreatitis is rare but potentially serious adverse effects.[br][br]Nothing to Disclose: NH, MS, KO, YO, RS, RI, AY, SU, KK, GY 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1522 56 314 SAT-211 PO47-02 Saturday 253 2012


254 ENDO12L_SAT-212 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Effect of Vildagliptin on Central Aortic Blood Pressure in Diabetic and Hypertensive Individual Luciana Neves Cosenso-Martin, Jose Fernando Vilela-Martin, Luiz Tadeu Giollo-Junior State Medical School of Sao Jose do Rio Preto (FAMERP), Sao Jose do Rio Preto, Brazil Background: Vildagliptin is a incretin-based therapy for the treatment of the type 2 diabetes (DM). Incretins are gastrointestinal hormones, predominantly glucagon-like peptide 1 (GLP-1), which are inactived by dipeptidyl peptidase-4 (DPP-4), and they stimulate insulin secretion. Vildagliptin is a DPP-4 inhibitor, improving [beta]-cell function, and decreasing glucagon secretion. GLP-1 has vasodilatory actions, and improves endothelium function in animals and humans. Thus, the objective of this case report was to evaluate the effect of the vildagliptin in the endothelium using the applanation tonometry of the radial artery (AT). [br]Clinical Case: Woman, 51 years-old, caucasian. She related DM for 4 years and high blood pressure for 6 years. Medicines in use: metformin 1700 mg/day, pioglitazone 30 mg/day, sinvastatin 10 mg/day, enalapril 5 mg/day and amlodipine 10 mg/day. No smoking. No DM in the family. At examination: Blood Pressure (BP): 129X81 mmHg; Heart rate: 78 bpm; BMI: 30 kg/m2; waist circumference: 91 cm[br]Glycated hemoglobina (HbA1c): 11,11%; Microalbuminuria: normal.[br]Evaluation of the central BP and Augmentation Index (AI) by AT were made before and after treatment with vildagliptin 100 mg/day. After 3 months, the patient returned taking vildagliptin and the medicines above described. No adverse effects were related. BP: 100X70 mmHg. HbA1c: 7,2%. AI and systolic central aortic BP reduced, 96% to 72% and 127 to 101 mmHg, respectively.[br]Clinical Lessions: AT is a noninvasive technique used to examine arterial elastic properties. An increased AI is associated with higher cardiovascular risk, and it was reported in patients with DM or impaired glucose tolerance. Higher values of AI indicate increased arterial stiffness. In health young individual, the systolic central aortic BP is around 20 mmHg below the systolic peripheric BP. Without vildagliptin the systolic central BP was only 2 mmHg lower than peripheric BP. This fact could be considered as an endothelium dysfunction, which was reversed by the vildagliptin, demonstrated by AI and systolic central BP reduction. Lower values for AI indicate a good elasticity of the arterial wall. We think this effect is not exclusive of the vildagliptin, because glucose control also can improve endothelium function. Others studies are necessary to confirm our results.[br]Conclusion: This case report is interesting because evaluate the central BP, demonstrating the effect of a DPP-4 inhibitor in endothelium dysfunction.[br][br]Nothing to Disclose: LNC-M, JFV-M, LTG-J 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1317 56 315 SAT-212 PO47-02 Saturday 254 2012


255 ENDO12L_SAT-213 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Puberty Onset of Diabetes Mellitus with 49, XXXXY Syndrome Takamasa Ichijo, Fumitaka Saida, Eiko Yoshida, Kaoru Yamashita, Hiromi Ouchi, Mariko Higa Saiseikai Yokohamashi Tobu Hospital, Yokohama, Japan [bold]Backbround: [/bold]The 49, XXXXY syndrome is a rare X-chromosome polysomy characterized by mental retardation, skeletal defects, craniofacial anomalies and hypogonadism (1). This syndrome is considered the most severe variant of Klinefelter syndrome. Although the incidence of diabetes among patients with Klinefelter syndrome or other types of X-chromosome polysomy is higher (2), the literature reported about puberty development of diabetes with 49, XXXXY syndrome are rare and only 2 cases have been reported in our knowledge. One possibility of this rare is most patients with 49, XXXXY syndrome are diagnosed in the newborn period rather than in adulthood as usually seen in 47, XXY Klinefelter syndrome.[br][bold]Clinical Case: [/bold]A 24-year-old man was admitted to our hospital for the treatment of hyperglycemia. He was diagnosed as 49, XXXXY syndrome at his birth and diabetes at 18-year-old. He was first treated by 1mg of glimepiride and increased the dose, but his plasma glucose level was not well controlled, and insulin injection therapy was started at 20-year-old. When he admitted to our hospital, he was treated by 30% insulin aspart premix twice a day, 24U at breakfast and 22U at supper, and his fasting glucose level was 143mg/dl and HbA1c was 7.9%. His height, weight and body mass index were 173.6 cm, 64.6 kg, and 21.4, respectively. His sexual development was Tanner stage I and small testes were palpable in his scrotum. His endogenous insulin profile, the fasting plasma c-peptide (CPR), 24 H urine CPR, and the glucagon test, showed 3.9 ng/ml (normal: [gt]0.5 ng/ml), 62.2 [mu]g/day (normal: [gt]20 [mu]g/day), and DCPR 1.2 ng/ml (normal: [gt]1.0 ng/ml), and his insulin secretion was conserved. His sex hormonal tests showed LH 13.0 mIU/ml (normal: 0.8-5.7 mIU/ml), FSH 40.8 mIU/ml (normal: 2-8.3 mIU/ml), and serum free testosterone [lt]0.5 pg/ml (normal: 2.55-7.53 pg/ml). We adjusted his administrating insulin and discharged with 22U of NPH at breakfast and 20U of 30% insulin aspart premix at supper, and his fasting glucose level and HbA1c were 112 mg/dl and 6.4% after a month. Because of his mother[apos]s refusal, the testosterone replacement therapy has not been started.[br][bold]Conclusion:[/bold] This is the third case report of 49, XXXXY syndrome with diabetes. We rather insist a periodical follow up is important to detect the development of diabetes and prevent the complications. We must refocus on his/her quality of life including prevention of diabetic complications in patients with 49,XXXXY syndrome.[br][br](1) Horm Res 2006;65;14-17. (2) J Clin Endocrinol Metab 1969;29;1062-1073.[br][br]Nothing to Disclose: TI, FS, EY, KY, HO, MH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 182 56 316 SAT-213 PO47-02 Saturday 255 2012


256 ENDO12L_SAT-214 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Successful Treatment of Acute Esophageal Necrosis Associated with Diabetic Ketoacidosis in a Patient with Type 1 Diabetes Maiko Mouri, Tsuguka Shiwa, Masayasu Yoneda, Tomokazu Awaya, Shuhei Nakanishi Chugoku Rosai Hospital, Kure, Japan; Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan A 64-year-old Japanese man was admitted to our hospital due to frequent vomiting and poor appetite for 2 days. He was diagnosed as diabetes 9 years ago and had been treated with biphasic insulin twice a day. He used to drink a lot and had drunk too much before his vomiting began. On admission, laboratory findings showed hyperglycemia (1,108 mg/dl) and metabolic acidosis, pH 7.215 with low bicarbonate levels. Ketone bodies in urine were elevated. These clinical findings indicated the diabetic ketoacidosis. Further, he presented melena. Upper gastrointestinal endoscopic examination showed black esophageal mucosa of the distal esophagus and erosion from just below the pharyngoesophageal constriction to the esophagogastric junction because of the presence of melena. Thus, he was diagnosed as diabetic ketoacidosis accompanying acute esophageal necrosis (AEN). He was immediately treated with continuous insulin administration to properly control blood glucose level as well as with total parenteral nutrition, oral intake restriction and intravenous administration of proton pump inhibitor. AEN was successfully treated without any complications. He was diagnosed as type 1 diabetes based on the following data. Anti-GAD antibody was positive (3.1 U/ml, normal range 0-1.4 U/ml), fasting serum C-peptide level was 0.1 ng/ml (normal range 0.8-2.5 ng/ml), and urinary C-peptide level was 0.4 [mu]g/day (normal range 22.8-155.2 [mu]g/day). After his recovery from AEN, his previous insulin therapy was replaced to basal-bolus insulin therapy to control the blood glucose level. AEN is a rare and life-threatening disorder. AEN usually develops to the perforation of esophagus, esophageal stricture, and superinfection of mucosa. The prognosis of AEN is largely dependent on the underlying disease, and the mortality rate approximately approaches to more than 30%. The immediate identification of AEN and the accurate management of blood glucose level with continuous insulin administration may lead to successful treatment of AEN without any complications. Taken together, AEN should be considered as one of the differential diagnoses of upper gastrointestinal bleeding in especially patients with diabetic ketoacidosis.[br][br]Nothing to Disclose: MM, TS, MY, TA, SN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 161 56 317 SAT-214 PO47-02 Saturday 256 2012


257 ENDO12L_SAT-215 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Use of Calcium-Stimulated Venous Sampling in Nesidioblastosis So-Young Kim, Ashutosh Pareek, Michael Via Beth Israel Medical Center, New York, NY; Beth Israel Medical Center, New York, NY Objective: To describe the utility of calcium stimulated venous sampling in suspected nesidioblastosis.[br]Case: A 36-year-old obese woman with a history of type 2 diabetes (DM2) and Roux-en-Y gastric bypass (RYGB) surgery five years earlier presented with syncope episodes she described as [ldquo]blackouts[rdquo] for 3 months, occurring at least three times a week. The episodes lasted less than one minute without any warning symptoms or signs of seizure-like activity. During one episode, the patient measured a finger-stick glucose (FSG) of less than 30 mg/dL. All DM2 medications were stopped, however, her episodic symptoms continued.[br]She was admitted to our institution for further evaluation and was given a 5% dextrose infusion. The following day, she again had symptomatic hypoglycemia with FSG of 43 mg/dL that improved with a 25 g dextrose infusion. At this time, patient had elevated serum insulin (6.8 uIU/mL, n[lt]3 uIU/ml), low pro-insulin ([lt]5pmol/L, n[lt]5pmol/L), low c-peptide (0.29 ng/mL, n[lt]0.69 ng/mL), non-detected sulfonylurea, insulin antibody [lt]0.4 U/mL, and low beta-hydroxybutyrate (38 mcg/mL, n[gt]280.8 mcg/mL). Computed tomography of abdomen/pelvis and magnetic resonance imaging of the brain did not demonstrate any masses. She produced a normal response to cosyntropin stimulation. A calcium localization test demonstrated a 2- to 3-fold increase of the insulin levels in the celiac, superior mesenteric, gastroduodenal, splenic, and hepatic artery. The patient was started on acarbose with meals and did not experience any further hypoglycemic episodes. She was subsequently given a 2-month course of parenteral nutrition for suspected malabsorption. She remained euglycemic after the parenteral nutrition was discontinued.[br]Discussion: Nesidioblastosis is an uncommon cause of hypoglycemia that is increasingly recognized following RYGB surgery. The diagnosis is often made clinically and confirmed histologically. In most cases, however, pancreatic surgery is not required. Arterial infusion of calcium followed by venous sampling can aid in the diagnosis of this condition.[br]Conclusion: In patients with prior RYGB surgery, the diagnosis of nesidioblastosis as a cause for episodic hypoglycemia may be established through calcium localization testing.[br][br]Nothing to Disclose: S-YK, AP, MV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 38 56 318 SAT-215 PO47-02 Saturday 257 2012


258 ENDO12L_SAT-216 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Use of Insulin Pump in Patients with Cystic Fibrosis-Related Diabetes: A Case Series Saket Gupta, Ronan Canavan St Vincent[apos]s University Hospital, Dublin, Ireland Cystic fibrosis related diabetes (CFRD); the most common complication of cystic fibrosis (CF) is associated with 6-fold increase in morbidity [amp] mortality. At present insulin is the only recommended medical treatment for CFRD (1) and multiple insulin injections (MDI) is the most effective regimen (2). Many CF patients don[apos]t adhere to regular insulin injections. A number of case reports and a small non [ndash] randomised study has reported improvement of glycaemic control and quality of life using continuous subcutaneous insulin infusion (CSII) via an insulin pump in the CFRD patients (3). However, there are no guidelines regarding the use of insulin pump therapy as a cost-effective treatment for CFRD.[br]We report our experience of CSII use in a series of CFRD patients at our institute.[br]The first patient, 18 year female, diagnosed with CFRD at age 15, was commenced on MDI in view of catabolic state (BMI 19 kg/m2) and poor glycaemic control (HbA1c 9.3%). She was poorly compliant to MDI and her nutritional status continued to decline despite PEG feed and subsequently CSII was commenced. During a 2-year follow on CSII, BMI increased to 24 kg/m2 however HbA1c failed to improve while FEV1 (69% of predicted) remained stable.[br]The second patient, 18 years male, diagnosed with CFRD at early age required insulin pump therapy because of declining lung function (FEV1, 50% of predicted) and malnutrition (BMI 18.2 kg/m2). The management of CFRD was complicated by his underlying decompensating liver disease. Intermittently he required hospital admissions for the management of complications related to liver disease. Despite serious illness, his BMI remained stable (19 kg/m2) and HbA1c fell (7.4% to 5.5 %). He subsequently underwent successful liver transplantation.[br]Another patient passed away within two years of commencing CSII of end-stage respiratory disease.[br]In summary, the use of CSII in our patients with CFRD resulted in improvement in the metabolic control nutritional status, and stabilisation of lung function. All three patients tolerated CSII with no pump related problems. Use of CSII in patients with CFRD has potential benefits in this complicated group of patients. Previous reports and our experience show that CSII may be a safe [amp] effective therapeutic alternative to MDI treatment in CFRD patients. However indications of CSII in CFRD should be based on individual patient requirement.[br][br]1. Moran A, et al., Diabetes Res Clin Pract 1999; 45: 61[ndash]73. 2. Borowitz D et al., J Pediatr Gastroenterol Nutr. 2002 Sep;35(3):246-59. 3. Hardin DS et al., J Cyst Fibros 2009 May;8(3):174-8. Epub 2009 Jan 21.[br][br]Nothing to Disclose: SG, RC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2081 56 319 SAT-216 PO47-02 Saturday 258 2012


259 ENDO12L_SAT-217 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Insulin Autoimmune Syndrome Mimicking Insulinoma: A Challenging Diagnosis Francesca Lugli, Donato Iacovazzo, Alessandra Fusco, Antonio Bianchi, Domenico Milardi, Sabrina Chiloiro, Marilda Mormando, Linda Tartaglione, Serena Piacentini, Andrea Giaccari, Alfredo Pontecorvi, Laura De Marinis Policlinico [quot]A Gemelli[quot] - Catholic University, Rome, Italy Insulin autoimmune syndrome (IAS) is a rare condition characterized by hypoglycemia due to autoantibodies against endogenous insulin. It is correlated, in approximately 50% of cases, to specific drugs intake. Hypoglycemia occurs typically during the late post-prandial period.[br]We report 2 cases of IAS patients who came to our attention because of recurrent and severe hypoglycemia.[br]Case 1: a 29 years old female, affected by Graves disease treated with methimazole, was evaluated in 2006 because of recurrent hypoglycemia. Insulinemia was [gt]1000 [micro]UI/ml. Fasting test was negative as well abdomen MRI. After total thyroidectomy glycemic and insulin levels normalized.[br]Case 2: a 45 years old female came to our attention in 2011 because of recurrent post-prandial hypoglycemia. Insulinemia was [gt]1000 [micro]UI/ml. An OGTT revealed late reactive hypoglycemia, while fasting test was normal as well EUS. Dosing of insulin antibodies was 658 U/ml ([lt]0.4). With an adequate diet and acarbose treatment the symptoms improved.[br]In these 2 cases, inappropriately high levels of insulin were associated with late post-prandial hypoglycemia with no evidence of pancreatic lesions and negative fasting tests.[br]In the evaluation of patients with hypoglycemia and inappropriate secretion of insulin, IAS should be considered in the differential diagnosis, especially in patients with late post-prandial hypoglycemia with history of specific drugs intake and negative imaging.[br][br]Nothing to Disclose: FL, DI, AF, AB, DM, SC, MM, LT, SP, AG, AP, LDM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1514 56 320 SAT-217 PO47-02 Saturday 259 2012


260 ENDO12L_SAT-218 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Clinical Characteristics of Fifteen Cases of Fulminant Type 1 Diabetes Mellitus in Our Hospitals Mariko Sue, Aya Yoshihara, Ryo Iga, Shuki Usui, Masahiko Miyagi, Mayumi Ishikawa, Yasuyo Ando, Hiromi Ouchi, Naoki Hiroi, Mariko Higa, Gen Yoshino Toho University School of Medicine, Ota-ku, Japan; Saiseikai Yokohama-shi Tobu Hospital, Tsurumi-ku, Japan [lt]Introduction[gt][br]Fulminant type 1 diabetes mellitus (FT1DM) is characterized by the remarkable acute onset and rapid development of ketoacidosis with nearly total destruction of pancreas b cells, whose etiology is hypothesized to be related to viral infection. We present clinical characteristics of 15 cases of FT1DM we experienced in our hospitals.[br][lt]Cases[gt][br]5 were women and 10 were men. The average age was 42.3 [plusmn] 15.5 year old.[br][lt]Results[gt][br]14 cases revealed acute onset with flu-like and/or abdominal symptoms. The average duration of onset was 4.5 [plusmn] 2.8 days. There was a discrepancy between extremely high plasma glucose concentration (1259 [plusmn] 574 mg/dL) and low levels of HbA1c (6.3 [plusmn] 0.5%). In addition, the serum and urine CPR levels were 0.1 [plusmn] 0.1 ng/mL and 2.4 [plusmn] 3.6 [mu]g/day, respectively, reflecting the severely damaged insulin secretion.[br]Even the relationship between FT1DM and viral infection is frequently suggested, the reports of specific viral infection are few. Though 2 cases revealed elevated herpes simplex virus or measles virus IgM, they did not represent the symptoms related to these viral infections, nor were the DNA levels of viruses negative.[br]In most patients with FT1DM, auto-antibodies are undetectable. However, there also are a few reports of cases with anti GAD Ab positive, or the cases that revealed changes from Ab negative to positive. In our cases, 3 cases were anti-GAD Ab positive, including 2 cases in low titer, and 1 case whose anti-GAD Ab changed from negative to positive, and subsequently changed to negative.[br][lt]Discussion[gt][br]In the first report, the negative status of auto-antibody was one of the sub criteria of FT1DM with the hypothesis that viral infection dominantly plays role in the pathogenesis. However, as many cases with anti-GAD Ab positive are reported, there remains the possibility that some part of autoimmune process contributes to FT1DM.[br]The absence of abnormality in imaging is one of the characters in FT1DM. Only 1 case described the swelling of pancreas in abdominal CT scan, which led to the misdiagnosis of acute pancreatitis. Including this, there were 2 cases misdiagnosed as gastritis or pancreatitis, whose plasma glucose concentration elevated to 1009 and 2230 mg/dL by the administration of inappropriate treatment, including intravenous infusion containing glucose. Though the concept of FT1DM has been accepted more recently, the mistake or delay in diagnosis are not rare even in Japan, where this disease was first reported.[br][br]Nothing to Disclose: MS, AY, RI, SU, MM, MI, YA, HO, NH, MH, GY 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 145 56 321 SAT-218 PO47-02 Saturday 260 2012


261 ENDO12L_SAT-219 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Dunnigan-Type Familial Partial Lipodystrophy [mdash] Case Report Sandra Belo, Angela Magalhaes, Paula Freitas, Cristina Gamboa, Davide Carvalho Centro Hospitalar de S[atilde]o Jo[atilde]o, Porto, Portugal; Centro Hospitalar de S[atilde]o Jo[atilde]o, Porto, Portugal; Universidade do Porto, Porto, Portugal [bold]INTRODUCTION:[/bold] Familial partial lipodystrophy, Dunnigan variety (FPLD), is a well recognized autosomal dominant disorder due to heterozygous missense mutations in lamin A/C (LMNA) gene. The Dunnigan-type familial partial lipodystrophy is characterized by a variable loss of fat in the extremities and trunk and excess subcutaneous fat in the chin and supraclavicular area. Associated metabolic abnormalities include low leptin, insulin resistance and dyslipidemia. [bold]CASE:[/bold] Female, 24 years old, past medical history of acute pancreatitis (1st episode October 2007, complicated with thrombosis of the superior mesenteric vein), combined dyslipidemia, diagnosed in this context, and diabetes, diagnosed in 2009, at first classified as secondary to pancreatitis. She had been referred to Endocrinology appointments but with irregular complacence. On August 2011 she was referred to the Pregnancy Endocrine pathology appointments in the context of 7 weeks pregnancy in a patient with diabetes. At physical examination she presented loss of fat in the extremities and trunk and excess subcutaneous fat in the chin. Weight 58 kilograms. At the first visit the therapy was insulin (initial total dose 106U), heparin and salmon oil. She presented good glycaemic control and triglycerides 1566 mg/dL ([lt]150mg/dL). Anti-GAD antibodies had been already requested and were positive. Dunnigan lipodystrophy was suspected and genetic study was requested. In September 2011 the patient was admitted to the emergency department for acute pancreatitis. Triglycerides were 9975mg/dL. As an inpatient, after improvement of TG with alimentary pause, therapy with fenofibrate was begun. She was then discharged. In October she presented TG of 3000mg/mL. She was then admitted for metabolic control. Therapy was then begun also with omega-3 polyunsaturated fatty acids and concentrated protein supplement after which TG were always around 1000-2000mg/dL. Genetic study revealed the presence of c.1444C[gt]T (p.Arg482Trp) heterozygote mutation in LMNA gene. The patient is now 34 weeks pregnant. [bold]CONCLUSION:[/bold] It is important to recognize the clinical features of this rare lipodystrophic syndrome due to its potentially severe consequences. This patient may benefit, after delivery, from leptin therapy.[br][br]Nothing to Disclose: SB, AM, PF, CG, DC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2200 56 322 SAT-219 PO47-02 Saturday 261 2012


262 ENDO12L_SAT-220 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Deep Neck Infection in a Diabetic Patient Joana Menezes, Elisabete Rodrigues, Jose Filipe Ramos, Hugo Guimaraes, Margarida Santos, Davide Carvalho Centro Hospitalar S[atilde]o Jo[atilde]o, Porto, Portugal; Centro Hospitalar S[atilde]o Jo[atilde]o, Porto, Portugal Introduction: Deep neck infections may be lethal if life-threatening complications occur, such as descending mediastinitis, pleural empyema, pericardial effusion, epidural abscess, adult respiratory distress syndrome and septic shock, especially in immunocompromised hosts such as diabetic patients(1). The occurrence of deep neck infections has been declining since the advent of antibiotic therapy; however, they do still occur and represent challenging diagnostic and treatment problems(2).[br]Case-Report: A 63-year-old woman with type 2 diabetes mellitus diagnosed 10 years ago and treated with oral antidiabetic drugs (metformin 2000mg/day, acarbose 50mg/day and gliclazide MR 30mg/day), hypertension and dyslipidemia was admitted in Otolaryngology department with neck swelling and redness of the submentonian region, with one week of evolution. She complained of odynophagia, dysphagia and neck pain, without fever, chills, trismus or voices changes. Laboratory results revealed white blood cell count over 12000cells/mm[sup]3[/sup] CPR=430.7mg/L and HbA1c=9.2%. The infection source of deep neck infection was unclear. The patient underwent a neck CT scan that showed a right parapharyngeal abscess with 13mm diameter with slight compression of upper air way, extending up to the lower edge of the right lobe of the thyroid with a longitudinal extent of about 85mm. The abscess involved the right half of the hyoid bone, showing a maximum diameter at this level of 30mm. CT scan also demonstrated densification of soft tissues under the chin region, compatible with inflammatory changes and cervical bilateral lymph nodes. The patient started on insulin therapy with good metabolic control, on intravenous antibiotic therapy (gentamicin and clindamycin) and underwent surgical drainage of the abscess. The results of microbiologic culture of purulent collection from surgery were negative as well as Koch Bacillus test. She presented a good clinical course, with discharge 8 days after admission.[br]Conclusion: Complications of deep neck infections are often life threatening. Clinical evidence and early radiologic diagnosis provide valuable information in defining the origin, location and extension of neck infections. In patients with diabetes mellitus, the clinical course is more severe and there is a poorer prognosis. The combination of appropriate intravenous antibiotic therapy, surgical drainage, securing of airway and good metabolic control are recognized cornerstones of treatment(3).[br][br](1)Mu-Kuan Chen et al, Am J Otolaryngol 2000; 21:169. (2) Silvio Marra et al, Am J Otolaryngol 1996; 17(5): 287. (3) Salih Bakir et al, Am J Otolaryngol 2010; 33: 63.[br][br]Nothing to Disclose: JM, ER, JFR, HG, MS, DC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1415 56 323 SAT-220 PO47-02 Saturday 262 2012


263 ENDO12L_SAT-221 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Case of New-Onset Type 2 Diabetes Mellitus in a Young Bodybuilder with No Known Family History of the Disease Niyaz Gosmanov, Donny Wynn OUHSC, Harold Hamm Diabetes Center, Oklahoma City, OK; Oklahoma City VA Medical Center, Oklahoma City, OK We present a 41 yo male of Puerto-Rican descent with past history of hypertension (HTN) and hypertriglyceridemia (HTG) has initially presented with diabetes in January 2011 when he was admitted to the hospital for nausea, abdominal pain, weakness and was found to have blood glucose values (BG) in 400[apos]s mg/dl range, triglycerides (TG) of 3200 mg/dl range (with no biochemical evidence of pancreatitis), HgA1c of 11.1% and diagnosed with hyperosmolar hyperglycemic state. He was promptly started on NPH 5 units BID and metformin 500 mg tid and discharged from the hospital with BG readings in 200 mg/dl range. Follow up with PCP resulted in addition of Glyburide 10 mg po bid and titration of NPH up to 10 units twice. Five months after initial presentation, his BGs were ranging 100-150 mg/dl; he had additional testing showing negative anti-GAD or islet cell antibodies and c-peptide of 2.7, insulin of 13 while glucose was 199 mg/dl. He has resumed his bodybuilding routine, though has made substantial dietary modifications with significant reduction of his carbohydrate (CHO) intake resulting in 20 lb weight loss. Twelve months after initial presentation, A1c is 6.1% on Acarbose 100 mg pot tid, Metformin 1000mg bid and occasional regular insulin (for large protein-enriched powder mix routinely used for work-out).[br]It remains unclear which factor dominated in the disease development in the physically active individual with no known family history of diabetes. It might be critically important that the patient[apos]s ethnic background has predisposed him to limited ability to produce insulin on high CHO diet. Of interest, normalization of TG levels has paralleled improved of diabetes control. Patient[apos]s life style and possible use of steroids or growth hormone have been investigated and shown that his Testosterone levels (total and free) as well as IGF-1, LH and FSH were normal and ratios have not indicated signs of exogenous injections; we have incidentally found that patient[apos]s prolactin level was slightly elevated. Patient[apos]s report about four other peer bodybuilders developing diabetes on similar exercise and meal regimens indicated that additional educational effort is needed to increase awareness about possibility of Type 2 diabetes development even among individuals on high diet/high exercise regimen with advice to periodically have lab tests. We stress need for supervision of dietary supplementation content abundantly used by bodybuilders for recreation purposes.[br][br]1. Matthew J Geraci, Mario Cole and Peter Davis., Hum Exp Toxicol 2011 30: 2007.[br][br]Nothing to Disclose: NG, DW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1670 56 324 SAT-221 PO47-02 Saturday 263 2012


264 ENDO12L_SAT-222 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Long-Term Stabilization of Diabetic Nephropathy and CAD with Triple Therapy of T2 Diabetes with TZD, Metformin and Ramipril Lee Alice Goscin Largo Medical Center, Largo, FL Both ACE Inhibitors and ARBS have shown reduction in microalbuminuria in both type 1 DM and type 2 DM. The Lewis study (1) showed captopril reduced death, dialysis and transplantation in type1 Dm. ARBS had similar benefits in subjects with type2 DM. The Hope trial (2) showed 3577 people with type2 DM over 55 years old with a history of a previous cardiovascular event or at least one other risk factor, 10 mg of ramipril reduced cardiovascular events and mortality by [ndash] 25%. In patients with type 2 DM, hypertension, microalbuminuria and renal insufficiency (Creatinine [gt]1.5 mg/dl), ARBS have been shown to delay the progression of nephropathy. This 77 year old man with triple coronary bypass, 750 mg of Proteinuria and previous Creatinine of 1.5 complicated by hyperkalemia has remained with creatinine of 1.1 and the same proteinuria for 14 years by using triple therapy of rezulin and then rosiglitazone, metformin and ramipril.[br]A second patient, a 58 year old woman with metabolic syndrome, experienced improvement of her Creatinine of 1.5 to 1.1 when started cautiously on metformin 500mg per day. The postulated mechanism may be reduction of endothelial activation and coagulation such as sVCAM, sICAM, and v WF. Twenty years of clinical experience with metformin as a first line medication in spite of creatinine levels slightly higher than1.6 suggests more consideration of higher creatinine clearance is warranted. A third patient remained with the same level of macroproteinuria for 5 years by remaining on her TZD. Indeed the PROACTIVE study (3) showed decrease in death, nonsilent MIs and strokes were reduced by 16%. In addition TZD[apos]s are beneficial in treating NASH and NAFL. These specific three patients suggest more studies are needed with emphasis on starting population and timing as most experienced clinicians practice. Removing existing patients in the United States such as this man from life-saving rosiglitizone is not warranted.[br][br](1) Lewis, N. Engl J. Med.,1963;. (2) Lancet, 2000;. (3) Dormandy, JA et al 2005, Lancet;366: 1279-1289.[br][br]Nothing to Disclose: LAG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2377 56 325 SAT-222 PO47-02 Saturday 264 2012


265 ENDO12L_SAT-223 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) The Relation between Diabetes Children and Insulin-like Growth Factor 1 (IGF-1) Araz Omer Rasheed University/55, Sulaimani, Iraq Introduction:[br]Insulin-like growth factor 1 (IGF-1) also known as somatomedin C or mechano growth factor is a protein that in humans is encoded by the IGF1 gene.It plays an important role in childhood growth and continues to have anabolic effects in adults.[br]Background:[br]IGF-1 consists of 70 amino acids. IGF-1 is produced primarily by the liver as an endocrine hormone as well as in target tissues in a paracrine/autocrine fashion. Production is stimulated by growth hormone (GH) and can be retarded by under nutrition, growth hormone insensitivity.[br]There is evidence of a metabolic role for IGF-I in type 1 diabetes.[br]In conclusion, IGF-I had a direct effect on glucose and protein metabolism, which was maintained during the hyperinsulinemic euglycemic clamp. This suggests that IGF-I acts in concert with insulin and may have an important role in maintaining glucose homeostasis and protein metabolism in type 1 diabetes.[br]As its name indicates IGF-1, has properties similar to insulin, and it has been shown to improve blood sugar profiles in type 2 diabetics.IGF-1 administration has actually normalized the insulin resistance in a group of healthy volunteers. While in these short term studies IGF-1 therapy did not completely do away with the need for insulin, it did improve the control of blood sugar and so may help prevent the complications of diabetes.[br]Clinical Case:[br]In the present study 40 children were randomly selected at Sulaimani pediatrics hospital aged (1-15) years old and subjected to be tested. The children were divided into two three groups according to their ages [amp]sex [amp]sugar level and IGF-1 and regiments to be teasing.[br]Result:[br]In our result we observed that IGF-1 has no any relation with age[apos]s[amp]sex. The correlation between (duration diabetes was (0.096) and IGF-1) is not significant.[br]And the correlation between (Low level sugar and IGF-1 was (0.14)) and for (high sugar level was (0.191)) is not significant.[br]For the sex the correlation between (males and IGF-1 was (0.837)) and (female [amp]IGF-1 was (0.096)) is not significant.[br]and the Correlation between (Age and IGF-1) is not significant.[br]Conclusion:[br]The obtained result showed that there were no significant effects (age[apos]s[amp]sex) of IGF-1 while it has a powerful rule with abnormality of sugar level.[br][br]Nothing to Disclose: AOR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 118 56 326 SAT-223 PO47-02 Saturday 265 2012


266 ENDO12L_SAT-224 POSTER SESSION: Diabetes [amp] Insulin Resistance: Uncommon Case Reports (1:30 PM-3:30 PM) Tropical Diabetic Hand Syndrome [mdash] Case Report Paula PB Silva, Gabriella D Arcari, Lenira C Stella, Fadlo F Filho, Isabela TI Carvalho, Renata P Bacchin Beneficencia Portuguesa, S[atilde]o Paulo, Brazil Diabetes Mellitus classically correlates to injuries of the extremeties, including debilitating injuries of hands.Typical diabetic complications related to the duration of the ilness and glycemic control, include infections, Dupuytren[apos]s Contracture syndrome,carpal tunnel syndrome and finger on trigger, more prevalent if diabetics than the general population.Tropical diabetic hand syndrome (TDHS) is a complication that affects patients with DM around the topics. Despite the prevalence of TDHS is still unclear in the literature, is defined as any diabetic adult patient with cellulitis, infection and gangrene. A 36-year-old woman, with DM1 for 35 years, went to the hospital with a necrotic and ulcero-infected lesion in the second chirodactyl of the right hand. She had already an antecedent of amputation of distal phalanx of the third finger on the right hand, after an insect bite. Physical examination revealed blood pressure of 90x50 mmHg, heart beat frequency was of 110/min. A small ulcer on the right second and third metacarpal bones was very warm and red with a purulent secretion. Laboratory test revealed leukocytosis, blood glucose level was 783 mg/dl and her HbA1c was of 12,5%. Ultrasonography of right hand diagnosed osteomyelitis and abscess on the those affected fingers. After glycemic control, anthibiotic therapy and a surgery for excision of the distal phalanx of the finger, she went home. However, after some days, the patient complained of pain, edema on the site of surgery and had the amputation of her second finger metacarpal. TDHS a potentially serious, because it can progress rapidly to gangrene. The predisposition to TDHS is multifactorial and includes uncontrolled diabetes, neropathy and insulin therapy. Antimicrobial tratment should be instituted immediatly based on the culture of biopsy and normally is necessary incision and drainage of the area. Without rapid and agressive treatment can occur permanent incapacity, amputation or death. The TDHS ends up to be neglected, causing delay in forwarding to the hospital. Prevention strategies include education of patients nad professionals for the examination of the hands, and the importance of establishing quick treatment regardless of severity of the injury as soon as the onset of the symptoms.[br][br]F Shiavon et al, Reumatismo 2004;56 (3), 139-142. Papanna et al, J Diabetes and its complications; 2010, 154-162.[br][br]Nothing to Disclose: PPBS, GDA, LCS, FFF, ITIC, RPB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2128 56 327 SAT-224 PO47-02 Saturday 266 2012


267 ENDO12L_SAT-225 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Measurement of Aldosterone in Serum by Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry Yuesong Wang, Gabrielle D Gay, Matthew Gatcombe, Julianne Cook Botelho, Hubert W Vesper Centers for Disease Control and Prevention, Atlanta, GA Aldosterone is a mineralocorticoid hormone secreted from the adrenal zona glomerulosa. Aldosterone secretion has been linked to the pathogenesis of hypertension and related cardiovascular disease. It is a major regulator of body fluid and ion transport, such as sodium reabsorption and potassium excretion, at very low circulating levels.[br]The measurement of aldosterone in serum is used clinically to investigate primary and secondary aldosteronism, and other disorders. Interpretation of the aldosterone level is typically conducted simultaneously with the plasma renin, salt intake, and in the context of upright or supine position. Immunoassay-based methods for measuring aldosterone are widely used in research and clinical diagnosis. However, antibodies can have varying selectivity and poor inter-laboratory reproducibility when measuring aldosterone in serum which limits their use in patient care and public health activities. Liquid chromatography tandem mass spectrometry (LC-MS/MS) can provide highly specific measurements of aldosterone. Herein, a highly specific, high-throughput procedure for measuring aldosterone in human serum by high performance liquid chromatography coupled tandem mass spectrometry was developed.[br]Aldosterone in serum is isolated from the sample matrix using liquid-liquid extraction. Interfering compounds are separated from the analyte using high performance liquid chromatography. Aldosterone is then measured by mass spectrometry. Sample processing is performed in an automated manner using well plates. Sample IDs are scanned automatically and linked to the LC-MS files. The HPLC separation was performed on a C18 column with a gradient and the mobile phases of water and methanol. Aldosterone and its stable isotope labeled internal standard were measured by selected reaction monitoring (SRM) in negative-ion mode. Aldosterone is quantified using the ion transition m/z 359 to 189 and confirmed by the transition m/z 359 to 331. The method performance such as sensitivity, recovery, robustness, and accuracy was well validated.[br][br]Nothing to Disclose: YW, GDG, MG, JCB, HWV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2181 57 328 SAT-225 PO09-01 Saturday 267 2012


268 ENDO12L_SAT-226 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Ratio of Plasma Aldosterone to Plasma Renin (ARR) Using Two Fully Automated Chemiluminescence Immunoassays Jenny Manolopoulou, Alexandra Bennett, Philipp Grimminger, Evelyn Fischer, Anja Pallauf, Sven Diederich, Martin Reincke, Martin Bidlingmaier Immunodiagnostic Systems Ltd, Boldon, UK; Medizinische Klinik und Poliklinik IV, Munich, Germany; Endokrinologikm Berlin am Gerdanmenmarkt, Berlin, Germany Background: Primary aldosteronism (PA) is reported to occur in 5-10% of the hypertensive population with increased incidence observed in recent years with the introduction of screening using the aldosterone to renin ratio (ARR). Variability in results from commercially available assays means there is a need for method specific cut-off values. We present here preliminary clinical data of two novel chemiluminescence immunoassays for plasma aldosterone (PAC) and plasma direct renin (DR) concentrations on an automated analyzer (IDS-iSYS), comparing results to commonly used assays.[br]Methods: PAC and DR were measured on the IDS-iSYS (Boldon, UK) and compared to the Siemens Aldosterone radioimmunoassay (RIA) and Diasorin Liaison Renin (LSN). 20 healthy controls, 26 essential hypertensives (EH) and 36 PA patients were included. PA was defined by lack of PAC salt-load suppression to [lt]50pg/mL using the Siemens RIA. Mineralocorticoid receptor antagonists and b-blockers were excluded. Hypokalemia was controlled in all subjects. Our laboratory cut-off during screening for PA is 12 (Siemens PAC (ng/L)/Liaison DR (mU/L)).[br]Results: Inter-assay imprecision on the aldosterone iSYS at 68pg/ml - 310pg/ml was at 15% - 4.3%, intra-assay CVs were 12-4%. For the renin iSYS assay, at 10.9mU/L - 309mU/L inter-assay imprecision was at 15% - 3.5%. PAC in controls, EHs and PAs were (mean; 25-75%ile): Siemens: 157(84.2-196), 123(73.3-137), and 274(190-320)pg/ml; iSYS: 115 (50.5-182), 114(68.7-131), and 279(172-348) pg/ml. In the same groups for DR: LSN assay: 33(21.2-43.1), 62(8.9-82.2), and 7.9(2.0-9.5) mU/L; iSYS: 40.1(27.4-47.8), 77.9(13.6-110), and 14.1(4.9-13.7) mU/L. Good correlation was observed for the aldosterone, iSYS=0.947RIA + 0.71, R2=0.71 and renin assays, iSYS=1.057LSN [plusmn] 0.067, R2=0.77. Using our current cut-off for the RIA and LSN at 12, ROC analysis of this cohort gives a sensitivity of 96.7% and specificity of 85.1%. At this cut-off the iSYS assays provided 93.6% sensitivity, so that a lower cut-off at 6.3 would provide corresponding 96.7%/84.8% sensitivity/specificity.[br]Conclusions: Our data demonstrate that the new assays present a convenient practical alternative for the simultaneous measurement of PAC and DR on a single automated analyzer. Comparable discrimination between the three groups and the same diagnostic power could be provided with a lower method specific ARR cut-off. The availability of these automated assays will facilitate screening and diagnosis of PA.[br][br]Disclosures: MB: Study Investigator, Chiasma. Nothing to Disclose: JM, AB, PG, EF, AP, SD, MR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1052 57 329 SAT-226 PO09-01 Saturday 268 2012


269 ENDO12L_SAT-227 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Role of Adrenal Vein Sampling in Primary Aldosteronism. Impact of Different Diagnostic Criteria on Subtype Diagnosis Maria-Verena Cicala, Anna Patalano, Monica Salva, Diego Miotto, Beatrice Rubin, Raffaele Pezzani, Barbara Mariniello, Franco Mantero Endocrinology Unit, University of Padua, Padua, Italy; Radiology Unit, University of Padua, Padua, Italy In patients with primary aldosteronism (PA), adrenal vein sampling (AVS) is considered the gold standard to distinguish between unilateral and bilateral autonomous production of aldosterone, while diagnostic imaging tests by CT scan or MRI are useful but often inconclusive for the diagnosis of PA. To date agreement is lacking on the best criteria indicating successful cannulation and lateralization.[br]The aim of the study was to evaluate the impact of different diagnostic criteria for the successful cannulation and lateralization on subtype diagnosis and to compare the difference of the findings between adrenal CT scan and AVS.[br]Sixty-seven patients with confirmed PA underwent AVS. The different diagnosis of PA subtype reached using AVS data assessed by more permissive (type 1) and strict (type 2) criteria were compared. Al patients performed CT scan before AVS and imaging results were compared with results of AVSs (using both criteria).[br]Using Type 1 criteria AVSs were successful in 86,3% of patients, while they were successful in only 66,6% using type 2 criteria. Type 1 criteria also led to a higher rate of diagnosis of unilateral PA (85% of successful procedures) than type 2 (75%). There was considerable disparity in the diagnosis reached using the 2 different criteria, with a concordance in only 50% of patients. In conclusion more permissive criteria for successful cannulation and lateralization on AVS can lead to incorrect diagnosis and accordingly to inappropriate treatment options. In the selected group of patients with successful AVS, CT findings correlated with AVSs findings and correctly identified unilateral or bilateral disease in 57,9% of patients using type 1 criteria and in 47,7% using type 2 criteria. Finale diagnosis was based on histological results in 29 patients (43,9%) which underwent adrenalectomy based on AVSs findings. On the basis of CT findings alone 17,5% of patients from the first group and 31,8% of patients of the second group probably would have been incorrectly bypassed as candidates for adrenalectomy. CT scanning lacks sensitivity and specificity and should, therefore, be followed by AVS, which is the only reliable means of differentiating unilateral from bilateral PA and lateralizing APAs preoperatively. However, there are still controversies to be solved by large prospective studies on the criteria to adopt for defining the most appropriate cut off for both correct cannulation and lateralization.[br][br]Nothing to Disclose: M-CC, AP, MS, DM, BR, RP, BM, FM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1252 57 330 SAT-227 PO09-01 Saturday 269 2012


270 ENDO12L_SAT-228 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Evaluation of Reproducibility of Captopril Loading Test in Primary Aldosteronism Masataka Kudo, Ryo Morimoto, Yoshitsugu Iwakura, Yoshikiyo Ono, Ken Matsuda, Akira Sugawara, Sadayoshi Ito, Fumitoshi Satoh Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku University Graduate School of Medicine, Sendai, Japan [italic]Introduction[/italic]: The plasma aldosterone concentration: renin ratio (ARR) is widely used for the screening of primary aldosteronism (PA), and then the captopril loading test (CAPT) is widely adopted as a confirmatory test of PA. There are many reports concerning about the accuracy of CAPT in PA diagnosis but little is known about its reproducibility. We, therefore, investigated the reproducibility of the ARR in CAPT among PA patients. [italic]Method:[/italic] A total of 48 PA subjects (male: 20, female: 28) who had CAPT 2 times were extracted from our PA patients data base in this study. The first CAPT (50 mg PO) was performed as an outpatient procedure and diagnosed as PA (basal ARR[gt]20 and the ARR in CAPT[gt]20). Successively, adrenal vein sampling (AVS) was performed to ascertain whether unilateral or bilateral excessive aldosterone secretion. The Second CAPT was performed after AVS during hospital stay. Blood samples were obtained before captopril administration, after that at 60min, 90min and 120min. The reproducibility of the ARR in CAPT at each time was assessed with Passing and Bablok regression, coefficient of reproducibility, and Bland-Altman plots. [italic]Results:[/italic] 12/48 patients presented with unilateral excessive aldosterone secretion and were underwent adrenalectomy as aldosterone producing adenoma (APA), whereas 36/48 patients presented with bilateral. The ARR in CAPT at 60min and 90min showed a highly significant within-patient correlation (ARR 60min: r=0.85 P[lt]0.01, ARR 90min: r=0.71 P[lt]0.01) and reproducibility. The ARR at basal and 120min showed a slightly lower correlation (basal ARR:r=0.45, ARR 90min:r=0.52). Bland-Altman plot showed no proportional, magnitude-related, or absolute systematic error between the ARR. [italic]Conclusion:[/italic] It is already reported that the basal ARR was affected by many factors, e.g. posture, time of blood sampling, sodium intake, kinds of drugs, gender and some genetic factors. Nevertheless, the ARR in CAPT showed a good reproducibility. From the points of view of reproducibility, the ARR 60 min was best. These data support that CAPT is a useful confirmatory test of PA.[br][br](1)Gian Paolo Rossi et al.,Hypertension 2010; 55:83. (2)Mohsen Agharazii et al.,Hypertension 2001; 37:1440.[br][br]Nothing to Disclose: MK, RM, YI, YO, KM, AS, SI, FS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1535 57 331 SAT-228 PO09-01 Saturday 270 2012


271 ENDO12L_SAT-229 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Cosyntropin Stimulation Test for the Subtype Classification in Primary Aldosteronism Kazutaka Nanba, Tetsuya Tagami, Tamiko Tamanaha, Kanako Nakao, Yohei Ueda, Maiko Kakita, Mika Tsuiki, Takeshi Usui, Akira Shimatsu, Mitsuhide Naruse National Hospital Organization Kyoto Medical Center, Kyoto, Japan Background: Primary aldosteronism (PA) is the common cause of endocrine hypertension with the prevalence up to 3 to 10% of hypertensive patients. Although adrenal venous sampling (AVS) is the gold standard for subtype classification in PA, it is available only in limited specialized centers. The aim of the study was to investigate the clinical significance of cosyntropin (ACTH) stimulation test for subtype classification in PA. Patients and Methods: Sixty patients with PA who underwent ACTH stimulation test were studied. The subjects were diagnosed as having either unilateral (n=41) or bilateral PA (n=19) based upon AVS, adrenal scintigraphy, and/or adrenal surgery. We evaluated the diagnostic significance of ACTH stimulation test in differentiating unilateral PA from bilateral PA. Results: Peak PAC (P[lt]0.01) and peak PAC/cortisol (P[lt]0.05) after ACTH stimulation were significantly higher in patients with unilateral PA than those with bilateral PA. Receiver operating characteristic curve analysis showed a peak PAC value of 403 pg/ml had a sensitivity of 70.7% and specificity of 79.0%, and a value of 596 pg/ml had a sensitivity of 46.3% and specificity of 100%. A peak PAC/cortisol value of 19.7 (PAC, pg/ml; cortisol, mcg/dl) had a sensitivity of 58.5% and specificity of 89.5% and a value of 30.5 had a sensitivity of 26.8% and specificity of 100%. Conclusions: ACTH stimulation test could discriminate between unilateral and bilateral PA and is useful in selecting the patients who should undergo AVS for localization before surgery.[br][br]Nothing to Disclose: KN, TT, TT, KN, YU, MK, MT, TU, AS, MN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1449 57 332 SAT-229 PO09-01 Saturday 271 2012


272 ENDO12L_SAT-230 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Aldosterone Effect on Erythrocyte Membranes: Comparison with Patients Affected by Hyperaldosteronism and Controls Chiara Sabbadin, Luciana Bordin, Gabriella Dona, Leila Bakdounes, Cristina Fiore, Giulio Clari, Decio Armanini University of Padua, Padua, Italy; University of Padua, Padua, Italy Aldosterone action has been associated with both genomic and non-genomic mechanisms. Aldosterone induces rapid non genomic effects by rising intracellular calcium and cAMP contents, triggering Na[sup]+[/sup]/K[sup]+[/sup] channel exchangers and increasing the activities of protein kinases, such as MAPK and PKC, and NADPH-oxidase. NADPH-oxidase system is the main superoxide generator and is probably involved in the aldosterone induced pro-inflammatory action, maybe in parallel with genomic effects.[br]Taking into account the particular sensitiveness of erythrocytes to the oxidative assault generated by reactive oxygen species (ROS), and their peculiarity as a-nucleated cells, erythrocytes can be particularly useful in the study of the non-genomic effect of aldosterone. In addition, previous studies evidenced that both the association in high molecular weight aggregates (HMWA) and the Tyr-phosphorylation (Tyr-P) level of erythrocyte membrane band 3 can be used as parameters to monitor the redox status of these cells.[br]We studied 12 patients affected by hyperaldosteronism (HA) and 6 healthy controls (HC), evaluating band 3 HMWA formation and Tyr-P levels, both in baseline conditions and after diamide treatment. Diamide is a bland oxidant which is efficaciously utilized to evidence pre-existent membrane alterations induced by oxidative stress.[br]In affected patients, diamide-induced band 3 Tyr-P was higher than controls, with a average increase of 132[plusmn]33%. In addition, also formation of band 3 HMWA induced by diamide was much higher in patients compared to healthy subjects (321.9[plusmn]8.7 compared to 130[plusmn]2%). This was expected due to diamide-induced disulfide bond formation between cysteine residues of band 3. More interestingly, in basal conditions, i.e. in absence of diamide, HA patients showed a net increase in the HMWA content (183[plusmn]8%) compared to HC, thus confirming that membranes from patients were altered.[br]In conclusion, our study demonstrates that aldosterone induces oxidative-like stress in a-nucleated cells, such as erythrocytes, by altering membrane structure. It would be of great interest to investigate this mechanism, through which this compound can regulate this non-genomic effect.[br][br]Nothing to Disclose: CS, LB, GD, LB, CF, GC, DA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1274 57 333 SAT-230 PO09-01 Saturday 272 2012


273 ENDO12L_SAT-231 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Predicting Factors of the Postsurgical Renal Function in Primary Aldosteronism Kanako Nakao, Kazutaka Nanba, Tamiko Tamanaha, Mika Tsuiki, Tetsuya Tagami, Takashi Usui, Akira Shimatsu, Mitsuhide Naruse National Hospital Organization Kyoto Medical Center, Kyoto, Japan Primary aldosteronism (PA) is the most common cause of secondary hypertension. Although decline in renal function especially that experienced after adrenalectomy (ADX) has been demonstrated, details of the mechanism remain to be elucidated. Aim of the study was to investigate the factors predicting renal outcome after ADX in PA. Fourteen patients with PA and 4 patients with non-functioning adrenal tumor (NFT) as control were studied. Estimated glomerular filtration rate (eGFR), serum potassium, plasma aldosterone concentration (PAC), and plasma renin activity (PRA) before and 1 month after ADX were analyzed. The study was approved by the institutional ethical committee. Baseline characteristics (M[plusmn]SD, PA vs. NFT) were as follows: age; 48[plusmn]9 vs. 65[plusmn]9 (yrs.), systemic blood pressure; 139[plusmn]10 (estimated duration of hypertension: 11.1[plusmn]10.2 yrs.) vs. 114[plusmn]10 (mmHg), eGFR; 92.8[plusmn]24.8 vs. 75.3[plusmn]19.2 (ml/min/1.73 m2), serum potassium; 3.1[plusmn]0.9 vs. 4.4[plusmn]0.3(mEq/L), PRA; 0.3[plusmn]0.3 vs. 1.0[plusmn]0.6 (ng/ml/h), PAC; 342[plusmn]235 vs. 87[plusmn]35 (pg/mL), and ARR; 2216[plusmn]2712 vs. 131[plusmn]102. Twelve PA patients (85.7%) showed decline in eGFR after ADX (92.8[plusmn]24.8 to 71.9[plusmn]24.1, p=0.04). Post-ADX eGFR were correlated negatively to estimated duration of hypertension (p=0.02) and preoperative PAC (p=0.04). There was no significant correlation between the post-ADX eGFR and the patient[apos]s age, blood pressure, serum potassium, and PRA. Chronological change in post-ADX eGFR could be further investigated in 13 PA patients. It was found that eGFR was already declined 1 day after ADX but did not show significant change over the period of 1 year. By contrast to the change in PA patients, there was no change in eGFR in NFT patients after ADX. The present study clearly demonstrated that the post-ADX decline in eGFR is a common and specific complication in PA patients. Duration of hypertension and preoperative PAC are the most important factors predicting the post-ADX eGFR, while there was no further aggravation of the renal function over a period of 1 year. It is therefore concluded that early diagnosis with correction of hypertension and hyperaldosteronism is essential to prevent decline of renal function after ADX in PA.[br][br]Sources of Research Support: Research Grant from National Hospital Organization in Japan for the Clinical Collaborative Study on Primary Aldosteronism.[br][br]Nothing to Disclose: KN, KN, TT, MT, TT, TU, AS, MN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1531 57 334 SAT-231 PO09-01 Saturday 273 2012


274 ENDO12L_SAT-232 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Prospective Multi-Center Study of National Hospital Organization in Japan on the Verification of the Guideline for Primary Hyperaldosteronism (PHAS-J2) Mitsuhide Naruse, Yasuishi Miyazaki, Tsuyoshi Tanaka, Masatoshi Shimizu, Sumire Otani, Shirou Hata, Atsushi Ogo, Kazuya Yonezawa, Kazuo Yoshida, Mamoru Sawamura, Rika Araki, Makito Tanabe, Mika Tsuiki, Tomoko Suzuki, Takurou Shimbo, Study PHASJ2 NHO Kyoto Medical Center, Kyoto, Japan; Misato Kenwa Hospital, Misato, Japan; NHO Mie Chuou Medical Center, Tsu, Japan; NHO Kobe Medical Center, Kobe, Japan; Saitama National Hospital, Wako, Japan; Nagasaki Goto Chuoh Hospital, Goto, Japan; NHO Kyushu Medical Center, Fukuoka, Japan; NHO Hakodate Hospital, Hakodate, Japan; NHO Nagasaki Kawatana Medical Center, Kawatana, Japan; Nishigunma National Hospital, Shibukawa, Japan; National Mie Hospital, Tsu, Japan; NHO Kokura Medical Center, Kokura, Japan; National Center for Global Health and Medicine, Tokyo, Japan Primary aldosteronism (PA) is the major cause of secondary hypertension. The guideline for PA established by the Endocrine Society recommends case detection, confirmatory testing, and subtype classification as the essential process. Given that PA is very common in hypertension, the guideline should be verified in the general practice of hypertension in the city hospital. Aim of the study was to investigate the implementation of each step by the multi-center collaborative study of National Hospital Organization (NHO) in Japan (PHAS-J2). Total 25 NHO and collaborated city hospitals with beds from 280 to 700 participated into the study. The study was approved by the institutional ethical committee. Patients with hypertension (20 to 75 yrs.) were enrolled. Plasma aldosterone to renin activity ratio (ARR) with a cutoff of 20 was used for case detection and captopril test was used as a confirmatory testing. Captopril-positive patients were subjected to subtype classification with adrenal CT, 131I-adosterol scintigraphy, and adrenal venous sampling (AVS). Total 1360 patients were enrolled. Screening test was positive in 21% and captopril test was positive in 58% of the screening-positive patients. Collectively, prevalence of PA was 12.2% of the patients. Adrenal tumor was visualized only in 32% on CT. Implementation of each step was 85% in captopril test, 93% in adrenal CT, 14.9% in adrenal scintigraphy, and 39% in AVS. In Kyoto Medical Center, total 242 patients with hypertension were enrolled. Screening test was positive in 46.7% and captopril test was positive in 65.1% of the screening-positive patients, providing the prevalence of PA as 30.3%. Adrenal tumor was detected in 44.4% on CT. While captopril test and adrenal CT was implemented in 76.1% and 96.4%, respectively, AVS was done only in 25.0% by various reasons. The present study of the NHO in Japan demonstrated that the prevalence of PA with positive screening and confirmatory testing is as low as 10% of the hypertension. In contrast to the screening and confirmatory testing, however, implementation of the subtype classification with AVS was still not high enough. Whatever the reasons underlying the results of the verification, the process of subtype classification requires alternatives as well as further advance in AVS for the standardization of clinical practice of PA in hypertension.[br][br]Sources of Research Support: Research Grant from the National Hospital Organization Japan for the Clinical Collaborative Study of Primary Aldosteronism.[br][br]Nothing to Disclose: MN, YM, TT, MS, SO, SH, AO, KY, KY, MS, RA, MT, MT, TS, TS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1385 57 335 SAT-232 PO09-01 Saturday 274 2012


275 ENDO12L_SAT-233 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Subtype Classification by Computed Tomography and Adrenal Venous Sampling in Primary Aldosteronism Sachiko Tsukamoto Kawashima, Kazutaka Nanba, Kanako Nakao, Yohei Ueda, Maiko Kakita, Tamiko Tamanaha, Mika Tsuiki, Takeshi Usui, Tetsuya Tagami, Akira Shimatsu, Mitsuhide Naruse National Hospital Organization, Kyoto Medical Center, Kyoto, Japan [Background] Subtype classification is the key procedure in determining the therapeutic strategy of primary aldosteronism (PA). Although CT has spread to many hospitals, it doesn[apos]t indicate the function of tumor nor detect microadenoma. By contrast, although adrenal venous sampling (AVS) is the gold standard for subtype classification, it is invasive and available only in limited centers. [Objective] Aim of this study was to compare the subtype classification by CT and AVS and to investigate the influence of different criteria of AVS for lateralization. [Subjects and methods] Forty eight patients with PA proved by confirmatory tests were studied. CT was decided to be positive if the tumor size was [ge]7 mm in its diameter. AVS was performed with ACTH stimulation and 4 different criteria were used for laterality: 1) adrenal PAC[ge]14000pg/ml, 2) PAC ratio of both sides(A/A)[ge]4, 3) lateralized ratio (LR)[ge]4 or 2.6, and 4) contralateral ratio (CR) [lt]1, respectively. [Results] On adrenal CT, 26 patients showed a unilateral adenoma and 22 patients showed normal or equivocal findings. In the CT-positive group, subtype classification by AVS was unilateral in 16 (62%), bilateral in 7 (27%), and contralateral in 3 (11%) with PAC criteria, unilateral in 14 (54%) and bilateral in 12 (46%) with A/A criteria, unilateral in 11 (42%) and bilateral in 15 (58%) with LR criteria[ge]4, unilateral in 13 (50%) and bilateral in 13 (50%) with LR criteria[ge]2.6, and unilateral in 10 (38%) and bilateral in 16 (62%) with CR criteria. In the CT-negative group, subtype classification by AVS was bilateral in 15 (68%) and unilateral in 7 (32%) with PAC criteria, bilateral in 18 (82%) and unilateral in 4 (18%) with A/A criteria, bilateral in 19 (86%) and unilateral in 3 (14%) with LR criteria[ge]4, bilateral in 15 (68%) and unilateral in 7 (32%) with LR criteria[ge]2.6, and bilateral in 20 (91%) and unilateral in 2 (9%) with CR criteria. Numbers of the criteria (PAC, A/A, LR[ge]4, and CR) full-filled in each patient of CT-positive group varied in each patient: 1 (n=16, 61%), 2 (n=14, 54%), 3 (n=10, 39%), and 4 (n=10,39%). The concordant rate of adrenal CT with AVS differed depending on the AVS criteria used. [Conclusions] Concordance of the subtype classification by adrenal CT and AVS depends on its criteria, cut-off, and the number of criteria full-filled, leading to a reversal of the subtype. Subtype classification even by AVS needs further standardization in terms of its diagnostic criteria.[br][br]Sources of Research Support: Research Grant from National Hospital Organization for the collaborative clinical study on primary aldosteronism.[br][br]Nothing to Disclose: STK, KN, KN, YU, MK, TT, MT, TU, TT, AS, MN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1089 57 336 SAT-233 PO09-01 Saturday 275 2012


276 ENDO12L_SAT-234 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Reevaluation of ACTH Loading in Adrenal Venous Sampling for the Diagnosis of Primary Aldosteronism Akiyo Tanabe, Aya Tsumagari, Mika Tsuiki, Tsuyoshi Tajima, Shuji Sakai, Atsuhiro Ichihara Tokyo Women[apos]s Medical University, Tokyo, Japan; Tokyo Women[apos]s Medical University, Tokyo, Japan; Kyoto Medical Center, Kyoto, Japan [bold]Objectives: [/bold]Corticotropin (ACTH) loading is commonly used in adrenal venous sampling (AVS) for lateralization of primary aldosteronism (PA). The aim of the present study was to reevaluate the diagnostic utility of ACTH loading during AVS. [bold]Subjects and methods: [/bold]AVS data from 127 patients with PA were analyzed. ACTH was administrated by bolus injection (250 [mu]g) in 20 patients and by continuous infusion (125 [mu]g/h) in 107 patients. The adrenal vein (AV) to inferior vena cava (IVC) cortisol (C) ratio [gt]2 before ACTH and [gt]5 after ACTH were used to document successful cannulation of each AV. Unilateral aldosterone excess was confirmed with an aldosterone lateralization ratio (LR) [gt]3; where the affected adrenal AV plasma aldosterone concentration (PAC)/C ratio is divided by the contralateral adrenal AV PAC/C ratio. In addition, the absence of aldosterone secretory autonomy was documented when the contralateral PAC/C ratio in the AV was less than that in IVC (contralateral ratio).[br][bold]Results: [/bold]1) Success rates of AVS. The success rate in the right AV was 60% before and 83% after ACTH loading. The success rate in left AV was 87% before and 97% after ACTH. 2) The concordant rate of lateralization before and after ACTH loading. In the patients with successful bilateral AVS both before and after ACTH (n=68), concordance in lateralization was seen in 69% of the patients. 3) The aldosterone LR. In the patients with proven unilateral aldosterone-producing adenoma (APA) by surgery (n=51), the accuracy rate of AVS lateralization was not different before (82.4%) or after (82.4%) ACTH. A change from negative to positive aldosterone LR was seen in 7.8% of the patients and an increase of this ratio [gt]100% in 23.5% of the patients after ACTH. By contrast, a change from positive to negative aldosterone LR was seen in 7.8% of the patients and a decrease in this ratio by [gt]50% was found in 29.4% of the patients after ACTH. 4) The contralateral ratio. In patients with APA, the accuracy rate of the absence of aldosterone excess from the contralateral AV was increased from 72.5% before to 90.2% after ACTH. The change from negative to positive result was seen in 17.6% of the patients after ACTH; there were no changes from positive to negative results.[br][bold]Conclusions: [/bold]ACTH loading increases the cortisol concentration gradient from the AV to IVC; as a result, it becomes easy to confirm successful AVS. However ACTH loading does not always improve the diagnostic value of AVS in lateralization of PA.[br][br]Nothing to Disclose: AT, AT, MT, TT, SS, AI 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 718 57 337 SAT-234 PO09-01 Saturday 276 2012


277 ENDO12L_SAT-235 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Usefulness of Super-Selective Adrenal Venin Sampling for Detecting Tiny Adrenal Lesions in Primary Aldosteronism Masao Omura, Kohzoh Makita, Seishi Matsui, Jun Inoue, Maki Nagata, Kunio Yamaguchi, Yukio Kakuta, Yoko Matsuzawa, Jun Saito, Tetsuo Nishikawa Yokohama Rosai Hospital, Yokohama, Japan; Tokyo-kita Social Insurance Hospital, Tokyo, Japan; Yokohama Rosai Hospital, Yokohama, Japan; Yokohama Rosai Hospital, Yokohama, Japan; Yokohama Rosai Hospital, Yokohama, Japan Introduction: Primary aldosteronism (PA) is one of the common diseases among hypertensives, and aldosterone-producing adenoma (APA) is usually treated by unilateral adrenalectomy. We had recently developed a new method of supper-selective ACTH-stimulated adrenal venous sampling (SS-ACTH-AVS) for exactly detecting the main lesion(s) of excess aldosterone or cortisol production. Then we attempted to partially remove the unilateral lesion of hyperaldosteronism detected by SS-ACTH-AVS.[br]Methods:SS-ACTH-AVS is done by using a strip-tip type 2.2 Fr micro-catheter (Omura-Makita Catheter; Koshin Medical Inc. Japan), of which the tip possesses three-quarters hall for obtaining easily adrenal effluent at intra-adrenal tributary veins. SS-ACTH-AVS was performed in 50 patients with PA, demonstrating unilateral CT-detectable adrenal nodules (Ult-N). Adrenal effluents were sampled at more than 2 intra-adrenal first-degree tributary veins in each adrenal gland after ACTH stimulation. When concentration of aldosterone in effluent sampled at one of the tributary veins connecting to any nodule was [gt]1400ng/dl and the others were [lt] 1400ng/dl, Ult-N was diagnosed as APA. Then, the patients with APA were treated by laparoscopic unilateral partial resection of the Ult-N without removing adherent normal tissues. When aldosterone was [gt]1400ng/dl in all effluents sampled at tributaries of the unilateral adrenal, showing the presence of Ult-N and was [lt]1400ng/dl in all effluents sampled at each tributary of the contralateral adrenal gland, the patients were also diagnosed as unilateral APA and treated by unilateral total adrenalectomy.[br]Results: Thirty patients, showing Ult-N were exactly diagnosed as APA after functionally detecting the localization by SS-ACTH-AVS, and were treated by unilateral partial adrenalectomy. Pathological examination of resected adrenal glands demonstrated that APA was completely removed without destruction of the capsules nor removing normal tissues. One year after adrenalectomy, concentrations of aldosterone in peripheral blood samples were normalized in 30 patients treated by partial adrenalectomy as well as in 20 treated by total one.[br]Discussion: SS-ACTH-AVS is a quite new method for detecting and localizing tiny adrenal lesions, and is also promising for choosing how to remove the adrenal lesion inducing hyperaldosteroneima, such as unilateral partial and total adrenalectomy.[br][br]Nothing to Disclose: MO, KM, SM, JI, MN, KY, YK, YM, JS, TN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 881 57 338 SAT-235 PO09-01 Saturday 277 2012


278 ENDO12L_SAT-236 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Validation of the Confirmatory Tests for the Diagnosis of Primary Aldosteronism; an Institutional Experience Takeshi Nigawara, Takako Moriyama, Aya Sugiyama, Shingo Murasawa, Satoshi Yamagata, Yutaka Watanuki, Yuko Tsushima, Shinobu Takayasu, Ken Terui, Satoru Sakihara, Kazunori Kageyama, Toshihiro Suda Hirosaki University Graduate School of Medicine, Hirosaki, Japan The high incidence of primary aldosteronism (PA) within the hypertensive population is widely acknowledged, and a need for efficacious diagnostic procedures has become a topic of discussion. Among various confirmatory tests, upright-furosemide test (Up-Fur), captopril test (Cap) and saline test (Sal) are officially recommended by Japanese Ministry of Health, Welfare and Labor, and positivity in any two of these tests is considered to corroborate the diagnosis of PA. We validated the usefulness of these examinations in our series of patients, together with cosyntropin test ([mu]g iv) without dexamethasone suppression, which is different from the method advocated by Sonoyama et al. (JCEM 2011). Clinical data of one hundred and ninety patients examined in suspicion of PA (plasma renin activity [lt] 2.0 ng/ml/min and plasma aldosterone concentration (Aldo) [ge] 8.0 ng/dl) in our department from January 2008 to June 2011 was collected, and subjected to statistical analysis. For patients taking angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers, calcium channel blockers or alpha-adrenergic inhibitors were prescribed in place of the former antihypertensive agents, prior to endocrinological examinations. A post-injection (30 or 60 min) plasma aldosterone concentration (Aldo) [gt] 20 ng/dl or peak Aldo/cortisol ([mu]g/dl) ratio [gt] 0.85 was tentatively considered as positive for PA in Cos. The basal Aldo in PA group (n=146) was 17.0 [plusmn] 9.1 ng/dl, whereas that in non-PA group (n=28) was 13.0 [plusmn] 5.2 ng/dl. The sensitivity of Cos, Up-Fur, Cap and Sal was 98.6, 92.4, 79.0 and 61.0%, respectively; while the specificity was 25.0, 91.0, 100 and 100%, respectively. This demonstrated that the above criteria in Cos rendered good sensitivity but very poor specificity. Alteration of the criterion to peak Aldo [gt] 30 ng/dl gave specificity of 92.0%, but resulted in a significant decline of sensitivity. Although Cos has an advantage in high sensitivity and the low risk of adverse effects, it may have a limited diagnostic value as a single test. The most useful single examination proved to be Up-Fur, but it is not feasible in elderly patients who have locomotor disabilities. In conclusion, the choice of confirmatory tests for PA should be made according to the patient[apos]s status, in combination with Up-Fur, when possible.[br][br]Nothing to Disclose: TN, TM, AS, SM, SY, YW, YT, ST, KT, SS, KK, TS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2018 57 339 SAT-236 PO09-01 Saturday 278 2012


279 ENDO12L_SAT-237 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) The Analysis of 87 Cases with Primary Aldosteronism [mdash] Evaluation of the Confirmatory Tests and Therapeutic Effect Against Atherosclerosis Takamasa Ichijo, Eiko Yoshida, Ayumi Yoshifuji, Kaoru Yamashita, Hiromi Ouchi, Mariko Higa Saiseikai Yokohamashi Tobu Hospital, Yokohama, Japan It is known primary aldosteronism (PA) quickens the atherosclerosis even if the blood pressure is well controlled. Since the guideline was established, we analyzed the usefulness of the confirmatory tests and therapeutic effect.[br]We screened the PA suspicious patients and 87 of those showed ARR[ge]20, which we considered positive. We, then, performed the confirmatory tests, such as rapid ACTH stimulating, captopril challenge, upright plus furosemide, and saline infusion tests, to those patients to diagnose PA. When any of those confirmatory tests were positive, and when surgical treatment is practicable, the diagnosis unilateral hyperaldosteronism (UHA) or bilateral hyperaldosteronism (BHA) was made by ACTH loading-adrenal venous sampling (AVS) after CT scans. We, then, finally perform unilateral laparoscopic adrenalectomy upon AVS documented unilateral hyperaldosteronism. Otherwise, we administrated mineralcorticoid receptor blockers (MRBs), spinorolactone or eplerenone.[br]The mean age was 55.3[plusmn]13.6 years old including 35 males and 52 females, and the mean ARR was 69.1[plusmn]82.0. Twenty-seven and 31 cases out of 65 AVS performed cases showed plasma aldosterone concentration[ge]1,400 ng/dl in either or both side, respectively. We then calculated the lateral ratio by aldosterone/cortisol in higher side to the other lower side, and 18 cases showed the ratio [gt]2.6, which we consider the laterality positive. Our AVS results revealed the mean ARR of UHA was significantly higher than one of BHA, 83.5[plusmn]110.1 vs. 51.7[plusmn]31.2, respectively, as we reported.[br]We next analyzed the anti-atherosclerotic effect of adrenalectomy and MRBs administration by the intima media thickness (IMT) of cervical artery, the brachial-ankle pulse wave velocity (baPWV) and ankle brachial index (ABI) of legs among 20 treated patients followed more than 1 year. Five cases were performed adrenalectomy and 15 cases were administrated MRBs among the AVS performed patients. The annual change of these surrogate makers revealed the adrenalectomy was more effective than MRBs, -0.04 vs. +0.01 mm, -57.33 vs. -36.11 cm/s, -0.03 vs. -0.02, respectively.[br]In conclusion, our results confirmed the usefulness of the multiple measurement of ARR to distinct between UHA and BHA before proceeding to the confirmatory tests, and both adrenalectomy and MRBs administration were effective, not only for anti-hypertensive but anti-atherosclerotic effect, though the former showed better outcome.[br][br]Nothing to Disclose: TI, EY, AY, KY, HO, MH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 158 57 340 SAT-237 PO09-01 Saturday 279 2012


280 ENDO12L_SAT-238 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Long-Term Outcome Following Adrenalectomy of Primary Hyperaldosteronism Caused by Unilateral Adrenal Hyperplasia Shalini Dabbadi, Jason C Jacob, Faripour Forouhar, Beatriz Tendler, Steven Shichman, Dougald MacGillivray, David Miner, Carl Malchoff University of Connecticut Health Center, Farmington, CT; Hartford Hospital, Hartford, CT; University of Connecticut Health Center, Farmington, CT; Maine Medical Center, Portland, ME; Middlesex Hospital, Middletown, CT Introduction. Primary hyperaldosteronism (PA) is caused by aldosteronomas, bilateral adrenal hyperplasia of the adrenal glomerulosa (BAH), and less frequently by unilateral hyperplasia of the adrenal glomerulosa (UAH). It is not known if UAH is a distinct entity amenable to long-term surgical cure or if it is an asymmetric variant of BAH with a propensity for PA recurrence over time following adrenalectomy. Few studies have carefully examined the long-term outcome of UAH following adrenalectomy. Hypothesis. We test the hypothesis that adrenalectomy produces a long-term cure in patients with PA caused by UAH. Methods. At the University of Connecticut Health Center we performed an IRB approved retrospective analysis of PA patients who underwent simultaneous bilateral adrenal vein sampling and subsequent unilateral adrenalectomy (1996-2010). The pathology was reviewed to identify all patients with UAH. Cure was defined as a post-adrenalectomy serum potassium concentration [gt]4.0 mEq/L without supplementation plus either serum aldosterone concentration (SAC) [lt]7 ng/dl or SAC (ng/dl) to plasma renin activity (PRA) (ng/ml/h) ratio [lt]15. SAC and PRA were measured with the patient sitting, and dynamic testing was not required to confirm normalization of the renin-aldosterone system (RAS). Results. We identified 8 patients with UAH; 1 refused post-adrenalectomy evaluation. Of the 7 available patients, 6 were cured by adrenalectomy. Hypokalemia resolved in the single patient who was not cured by adrenalectomy, but other medical problems precluded safe dynamic testing of the RAS. One patient died from unrelated causes 2 y following adrenalectomy. Pathologically, the adrenal glands of all UAH patients demonstrated triangular glomerulosa hyperplasia with nodular architecture, and 4 had dominant nodules from 0.5 cm to 1.2 cm in diameter. The 6 cured patients were followed for a mean of 8.0 y (range = 2.0 y to 15.2 y) after adrenalectomy, and each remained cured of PA. Discussion. There are limited reports concerning the long-term outcome of UAH patients following adrenalectomy. The complete long-term resolution of all RAS abnormalities in 6 of 7 patients with UAH is most consistent with the hypothesis that UAH is usually a distinct entity and not an asymmetric variant of BAH. Conclusion. Approximately 85 % of UAH patients experience a long-term cure following adrenalectomy. UAH is likely a distinct entity and not an asymmetric variant of BAH.[br][br]Nothing to Disclose: SD, JCJ, FF, BT, SS, DM, DM, CM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 373 57 341 SAT-238 PO09-01 Saturday 280 2012


281 ENDO12L_SAT-239 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Serum Selenium Concentrations and Hypertension in the German Population Ioanna Gouni-Berthold, Bernhard Michalke, Wilhelm Krone, Eliseo Guallar, Heiner Berthold University of Cologne, Cologne, Germany; German Research Center for Environmental Health, Neuherberg, Germany; Johns Hopkins Bloomberg School of Public Health, Baltimore, MD; Research Groupon Geriatrics, Berlin, Germany Selenium is an essential element with antioxidant properties mediated through various selenoenzymes. Because oxidative stress is involved in hypertension development, it has been suggested that selenium may be involved in blood pressure control and hypertension prevention. The few studies that have investigated the association between serum selenium concentrations and blood pressure have been inconsistent, reporting inverse, null, or positive associations. We explored the relationship between serum selenium concentrations and blood pressure levels and hypertension in the German population. We undertook a cross-sectional analysis of 792 Caucasian subjects (46% male; mean [plusmn] SD age, 53 [plusmn]14 years) who participated in the Lipid Analytic Cologne (LIANCO) cohort. LIANCO was a cohort study designed to assess the relationships between genetic mutations, serum lipoproteins, other biochemical parameters, and clinical data in hypertension, diabetes and atherosclerotic disease. Hypertension was defined as having a systolic blood pressure [ge]140 and/or a diastolic blood pressure [ge]90 mmHg. About half of the cohort was diagnosed as hypertensive. Selenium was measured by inductively coupled plasma-dynamic reaction cell-mass spectrometry (ICP-DRC-MS). Mean [plusmn] SD serum selenium concentration was 68 [plusmn] 32 [mu]g/L, i.e about half of the concentrations seen in the US population. The multivariable adjusted (for sex, age, and body mass index) differences (95% confidence intervals) in blood pressure levels comparing the highest ([gt] 91.9 [mu]g/L) to the lowest ([lt]42.8 [mu]g/L) quartile of serum selenium were 5.2 (1.4 to 8.9), 2.8 (0.7 to 4.8), and 2.4 ([ndash]0.4 to 5.2) mmHg for systolic, diastolic, and pulse pressure, respectively ([italic]P[/italic] for trend for all [lt]0.003). The corresponding multivariable adjusted odds ratio for the presence of hypertension was 1.52 (0.98 to 2.36; [italic]P [/italic]for trend=0.004).[br]The present data suggest that even in a population with very low serum selenium concentrations higher serum selenium concentrations are associated with higher blood pressure levels and a higher prevalence of hypertension. These findings call for careful evaluation of the risks and benefits associated with high selenium concentrations, and may question the rational of the current widespread use of selenium supplements.[br][br]Nothing to Disclose: IG-B, BM, WK, EG, HB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 43 57 342 SAT-239 PO09-01 Saturday 281 2012


282 ENDO12L_SAT-240 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Low-Sodium Diet Is a Significant Predictor of Increased Catecholamine Levels in Normotensive and Hypertensive Men and Women Aditi R Saxena, Bindu Chamarthi, Ellen W Seely Brigham and Women[apos]s Hospital, Boston, MA Context: Many factors are proposed to affect catecholamine levels, including: age, gender, hypertensive disease and dietary sodium. However, no studies have examined relative effects of these factors within the same study population.[br]Objective: To determine what factors significantly predict both plasma and urinary catecholamine levels.[br]Methods: Hypertensive and normotensive subjects (n=308) were studied in high (250 mEq sodium per day) and low sodium balance (10 mEq of sodium per day). Antihypertensive medications were discontinued in all subjects two weeks before the study. Twenty-four hour urine collections were used to confirm sodium balance and to measure urinary norepinephrine, epinephrine, and creatinine. Plasma norepinephrine and epinephrine were measured in the morning after an overnight fast. Gender, race, age, body mass index (BMI), dietary salt exposure (high salt vs. low salt), hypertension status, and mean arterial pressure were demographic variables of interest. Repeated measures multivariate linear regression models were used to examine the effect of these covariates on plasma and urinary epinephrine and norepinephrine levels. Best-fit lines were obtained using the least-squares method.[br]Results: Of the variables listed above, only the following factors significantly predicted plasma norepinephrine levels: age ([beta]= 2.1, SE=0.8, p=0.005), Black race ([beta]=50.2, SE 23.0, p=0.03), and low salt diet ([beta]=64.9, SE=11.5, p[lt]0.0001). With respect to 24-hour urinary NE, there were a greater number of significant predictors: age ([beta]=0.5, SE=0.2, p=0.005), female gender ([beta]= -10.7, SE=3.5, p=0.002), BMI ([beta]=1.5, SE=0.4, p=0.0009), low salt diet ([beta]=16.4, SE=2.0, p[lt]0.0001). Race did not significantly predict urinary NE levels, controlling for other variables. In contrast, only female gender and low salt diet significantly predicted plasma epinephrine levels, with the following effects: female gender ([beta]= -6.5, SE=2.9, p=0.02) and low salt diet ([beta]=4.8, SE=2.4, p=0.05). With respect to 24-hour urinary epinephrine, only female gender was a significant predictor ([beta]= -4.1, SE=0.9, p[lt]0.0001).[br]Conclusion: In relation to other variables known to affect catecholamine levels, low dietary sodium is the most significant predictor of increased catecholamine levels. These results suggest that multiple factors affect plasma and urinary catecholamine levels, and these should be considered when interpreting catecholamine values in patient and study populations.[br][br]Nothing to Disclose: ARS, BC, EWS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1633 57 343 SAT-240 PO09-01 Saturday 282 2012


283 ENDO12L_SAT-241 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) High Salt Intake Is Associated with Increased Production of Cortisol, Aldosterone Metabolites and Metabolic Syndrome Rene Baudrand, Jaime Cerda, Carmen E Campino, Cristian A Carvajal, Macarena Jimenez, Daniela Figueroa, Fidel Allende, Sandra Solari, Oliviero Olivieri, Carlos F Lagos, Carolina Valdivia, Gareth Owen, Carlos E Fardella School of Medicine, Faculty of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile; School of Medicine, Faculty of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile; Millennium Institute of Immunology and Immunotherapy, Santiago, Chile; School of Medicine, Faculty of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile; University of Verona, Verona, Italy Background and Aims: Excess salt intake, highly prevalent worldwide, is associated with hypertension (HT), insulin resistance (IR) and cardiovascular events, but its physiopathology has not been fully elucidated. We evaluate a possible dysregulation of cortisol (F) and aldosterone (A) production secondary to high salt intake, that could have a key role in metabolic syndrome (MetS).[br]Subjects and Measurements: We recruited 370 adults (17-85 years, BMI 29.3 [plusmn] 4.4 kg/m2 ,70% women, 68% had HT, 43% had MetS by ATPIII criteria). High salt intake, defined by urinary sodium [gt]150 mEq/24h, was detected in 69% of recruited subjects. We evaluated in plasma: F, cortisone (E), A, renin activity (PRA), leptin, adiponectin, glucose, insulin and lipid profile. In 24h urine: F and E were measured by LC/MS-MS and tetrahydroaldosterone (THA) and F tetrahydrometabolites (THM) by GC/MS. IR was estimated by HOMA Calculator 2.2. Results are expressed as median [Q1-Q3].[br]Results: Subjects with high, compared to normal salt intake, presented higher values of urinary F (48.3 ug/24h [32.9-61.5] vs 29.2 ug/24h [20.2-37]; p=0.001), THM (8.1 mg/24h [6.2-10.7] vs 5.6 mg/24h [4-7.9]; p[lt]0.001), THA (80.2 ug/24h [50-110] vs 60 ug/24h [37.5-92.5]; p[lt]0.001), urinary F/E ratio (0.46 [0.3-0.6] vs 0.32 [0.2-0.5]; p=0.015), HOMA-IR (3.1 [1.5-5] vs 2.4 [1.1-3.9]; p=0.04) and a higher % of MetS (61 vs 39; p=0.004). We observed a positive correlation between salt intake and urinary F (r=+0.45,p[lt]0.001), THM (r=+0.41,p[lt]0.001), THA (r=+0.26,p[lt]0.001), urinary F/E ratio (r=+0.26,p=0.003) and BMI (r=+0.12,p=0.02) and a negative correlation with adiponectin (r=-0.17,p=0.004) and HDL-C (r=-0.15, p=0.007). By ANCOVA analysis, adjusted by age and BMI, high salt intake was associated with higher THM (p[lt]0.001) and HOMA-IR (p=0.01) and lower adiponectin (p=0.01). In adjusted logistic regression model, high salt intake was associated with MetS (OR= 2.2 [95% IC 1.2-3.8]).[br]Conclusions: In our cohort, high salt intake was associated with increased cortisol production, increased cortisol and aldosterone urinary metabolites and higher urinary F/E ratio. In addition, high salt intake was associated to insulin resistance, dyslipidemia, hypoadinectinemia and increased risk of metabolic syndrome, even after adjusting for confounding variables. Our results suggest that the association of high salt intake with metabolic disorders could be, at least partially, related to increased cortisol and aldosterone production.[br][br]Sources of Research Support: FONDEF D08I1087, Fondecyt 110036 and Millennium Institute in Immunology and Immunotherapy P09/016-F.[br][br]Nothing to Disclose: RB, JC, CEC, CAC, MJ, DF, FA, SS, OO, CFL, CV, GO, CEF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 582 57 344 SAT-241 PO09-01 Saturday 283 2012


284 ENDO12L_SAT-242 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) The Role of Serum Factor in the Pathogenesis of Obesity-Related Idiopathic Hyperaldosteronism Takako Ohyama, Hirotaka Shibata, Isao Kurihara, Ayano Murai-Takeda, Yuko Mitsuishi, Takeshi Hayashi, Rie Jo, Kazutoshi Miyashita, Hiroshi Itoh School of Medicine Keio University, Tokyo, Japan [bold][underline]Background:[/underline][/bold] Primary aldosteronism (PA) is characterized by hypertension, autonomous aldosterone secretion, and higher prevalence of metabolic syndrome. Two most common subtypes of PA are aldosterone-producing adenomas (APAs) and bilateral idiopathic hyperaldosteronism (IHA). The pathogenesis of APAs is due to autonomous overexpression of CYP11B2 or the [italic]KCNJ5[/italic] potassium channel gene mutation, whereas that of IHA is totally unknown at present. Previous reports showed that human [ldquo]adipocyte-derived factors[rdquo] stimulate aldosterone secretion in human adrenocortical cells. We therefore investigated whether the serum have stimulatory effects on aldosterone secretion in patients with IHA.[br][bold][underline]Patients and Methods:[/underline][/bold] (1) Correlation between urinary aldosterone excretion and metabolic parameters, such as BMI, HbA1c, glucose, HOMA-R, HDL-C, LDL-C, and TG were analyzed in patients with PA (APA, n=36; IHA, n=85) and non-PA patients (n=57). (2) The effects of patient[apos]s serum on the [italic]CYP11B2[/italic]-luciferase reporter activity in human adrenocoritcal H295R cells. The H295R conditioned medium contained 5% Nu-Serum and 20% patient[apos]s serum.[br][bold][underline]Results :[/underline][/bold] (1) The BMI (P=0.014), waist circumference (P=0.008), HOMA-R (P=0.003), and plasma glucose (P=0.005) values were significantly correlated with urinary aldosterone excretion in patients with IHA, but not those with APA or non-PA. (2) We next investigated whether addition of patient[apos]s serum into the culture medium affect the [italic]CYP11B2[/italic]-luciferase reporter activity. The results showed that patient[apos]s serum significantly increased the reporter activities by 1.9-fold, 1.9-fold, and 1.7-fold, in patients with IHA, APA, and non-PA, respectively. Of interest, when we investigated obese patients whose BMI value is greater than 25, co-treatment with patient[apos]s serum and protein kinase A inhibitor (H89) merely suppressed the [italic]CYP11B2[/italic] reporter activities by 6% in IHA, whereas significantly suppressed the activities by 43% and 77% in APA and non-PA, respectively.[br][bold][underline]Conclusion:[/underline][/bold] Clinical profiles strongly indicate that aldosterone excess in IHA may be associated with obesity and insulin resistance. The serum [italic]CYP11B2[/italic]-stimulating activity was confirmed in all patients; however, the activity was drastically cancelled with H89 treatment in APA and non-PA, but not IHA. These data suggest that PKA-independent serum factors which may be derived from adipocytes play crucial roles in excessive aldosterone secretion in patients with obesity-related IHA.[br][br]Nothing to Disclose: TO, HS, IK, AM-T, YM, TH, RJ, KM, HI 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 728 57 345 SAT-242 PO09-01 Saturday 284 2012


285 ENDO12L_SAT-243 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Vitamin D3 Therapy Lowers Tissue Renin-Angiotensin System Activity in Obesity: A Prospective Study Anand Vaidya, Bei Sun, Carol Larson, John P Forman, Justin P Annes, Jonathan S Williams Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA; Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA; Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA Background: Increased activity of the tissue renin-angiotensin system (RAS) is a leading mechanism for obesity-related hypertension (HTN) and chronic kidney disease (CKD). Low vitamin D levels are associated with increased RAS activity, the development of CKD and HTN, and are common in obesity. We hypothesized that vitamin D3 therapy in obesity could lower tissue-RAS activity and therefore raise the tissue-sensitivity to angiotensin II (AngII).[br]Methods: 19 obese subjects with HTN, 25(OH)D[lt]25 ng/mL, and normal renal function were enrolled; 14 completed all study procedures. Anti-hypertensive medications were stopped for up to 3 months prior to the 1st study visit to eliminate interference with the RAS, after which subjects received 1 month of vitamin D3 15,000 IU/day, and returned for a 2nd study visit. Controlled dietary sodium balance was maintained during both study visits. Tissue-RAS activity was assessed at both study visits by measuring the responses of mean arterial pressure (MAP), renal plasma flow (RPF), and adrenal aldosterone secretion to an infusion of AngII (1 ng/kg/min). Subjects were then treated with captopril, and underwent a second infusion of AngII to evaluate the change in tissue-RAS induced by captopril.[br]Results: Following vitamin D3 therapy, mean 25(OH)D and 1,25(OH)2D rose [18 to 52 ng/mL and 38 to 58 pg/mL respectively] and PTH fell [46 to 37 pg/mL] (P[lt]0.01 for all). Additionally, supine MAP declined [105.6 to 102.7 mmHg] and basal RPF increased [383.6 to 401.0 mL/min/1.73m2] (P[lt]0.01 for both). There was a greater decline in RPF and a higher mean stimulation of aldosterone with AngII infusion following vitamin D3 therapy (P[lt]0.05). As anticipated, captopril increased the RPF, MAP, and aldosterone responses to AngII, but this effect was much smaller at visit 2, indicating that vitamin D3 therapy increased the tissue-sensitivity to AngII akin to captopril (P[lt]0.05). Vitamin D3 therapy did not influence levels of TGF-[beta], PAI-1, or IL-6, and hypercalcemia or hypercalciuria were not detected.[br]Conclusions: In this controlled study, vitamin D3 therapy in obese hypertensives modified RPF, MAP, and the tissue-sensitivity to AngII similar to the action of a converting enzyme inhibitor, indicating that vitamin D3 therapy reduces tissue-RAS activity. Whether chronic vitamin D3 therapy in obesity abrogates the development of CKD and HTN via reducing RAS activity warrants investigation.[br][br]Sources of Research Support: F32 HL104776-02 (AV), K23 HL08236-03 (JSW).[br][br]Nothing to Disclose: AV, BS, CL, JPF, JPA, JSW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 40 57 346 SAT-243 PO09-01 Saturday 285 2012


286 ENDO12L_SAT-244 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Processing of Precursor Aldosterone Synthase by Mitochondrial Processing Peptidase Requires Precise Interaction in the Mitochondrial Matrix Brian P Adams, Himangshu S Bose Mercer University School of Medicine, Savannah, GA Majority of mitochondrial proteins are encoded by the nucleus, synthesized in the cytosol and directed to the mitochondria by their own peptide signaling sequences. Proteins targeted to the mitochondrial matrix have an N-terminal sequence that is cleaved after matrix translocation. Mitochondrial processing peptidase (MPP) cleaves precursor to the mature form by precise recognition of the protease cleavage site. For instance, inhibition of aldosterone synthase (AS) cleavage by MPP increased activity. MPP cleaves AS between the 24th and 25th amino acids. Mutation of the 24th amino acid from leucine to aspartate prevents MPP cleavage. To understand the mechanism of AS precursor processing, we focused on how the cleavage site of AS interacts with MPP. Based on the AS crystal structure we predicted interactions of AS residues with the MPP active site from amino acids 22 through 25 as critical for AS recognition. Mitochondrial import showed reduced cleavage of the mutants compared to wild-type. Mutations of arginine 22 to leucine, leucine 24 to proline, and glycine 25 to asparagine reduced cleavage of those mutants 50.0 [plusmn] 3.1%, 23.4 [plusmn] 2.4%, and 57.2 [plusmn] 1.7%, but were translocated into the mitochondrial matrix. Activity of the mutants is 20-90 times more than the wild type AS. Mutation of alanine 23 to serine had no effect on cleavage, but we found this mutant to be inactive. These results reveal a specific molecular interactions for the activity and mitochondrial cleavage of AS is necessary to regulate mineralocorticoid balance.[br][br]Sources of Research Support: National Institutes of Health HD057876.[br][br]Nothing to Disclose: BPA, HSB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1285 57 347 SAT-244 PO09-01 Saturday 286 2012


287 ENDO12L_SAT-245 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) High Glucose Stimulates Aldosterone Synthase Gene ([italic]CYP11B2[/italic]) Expression in Human Adrenal H295R Cells Takako Saito-Ito, Ken Matsuda, Akira Uruno, Kyoko Shimizu, Miki Nakamura, Rou Takahashi, Takeo Yoshikawa, Masataka Kudo, Akiko Saito-Hakoda, Sadayoshi Ito, Akira Sugawara Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku University Graduate School of Medicine, Sendai, Japan Introduction: Aldosterone synthase gene ([italic]CYP11B2[/italic]) is the key enzyme of adrenal aldosterone production, and the examination of its regulation is essential for the elucidation of the etiology of hypertension. We here examined the effects of high glucose on [italic]CYP11B2[/italic] transcription using a newly obtained stable H295R cell line expressing[italic] CYP11B2[/italic] 5[apos]-flanking region/luciferase cDNA chimeric construct. Methods: Human adrenal H295R cells and the stable H295R cell line were used. The cells were incubated with several concentrations of D-glucose. The cellular osmolarity was adjusted by L-glucose. mRNA expression of[italic] CYP11B2[/italic] and transcription factors was determined by real-time PCR. Aldosterone secretion from H295R cells to the media was measured by EIA. In some experiments, transient transfection studies were also performed. Results: D-glucose dose- and time-dependently stimulated [italic]CYP11B2[/italic] transcription. Moreover, high concentrations of D-glucose also stimulated[italic] CYP11B2 [/italic]mRNA expression and aldosterone secretion. Interestingly, high-glucose also stimulated the expression of transcription factors nerve growth factor-induced clone B (NGFIB) and Nur-related factor 1 (NURR1). The high-glucose stimulated [italic]CYP11B2[/italic] transcription was not affected by angiotensin II receptor blockers and LY333531 (a protein kinase C-b inhibitor) treatment, but was abrogated by treatment with KN-93, a Ca[sup]2+[/sup]/calmodulin-dependent kinase (CaMK) inhibitor. Transient transfection experiments using [italic]CYP11B2[/italic] 5[apos]-flanking region deletion mutants revealed the possible involvement of NBRE-1 element that is transactivated by NGFIB and NURR1. Conclusion: High-glucose was demonstrated to stimulate [italic]CYP11B2[/italic] transcription possibly via CaMK-mediated NBRE-1 element activation. Our observation may give us the clue for the elucidation of the link between hypertension and diabetes mellitus.[br][br]Nothing to Disclose: TS-I, KM, AU, KS, MN, RT, TY, MK, AS-H, SI, AS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1475 57 348 SAT-245 PO09-01 Saturday 287 2012


288 ENDO12L_SAT-246 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Angiotensin II Stimulation of Aldosterone Production Is Mediated by Kir3.4 in HAC15 Cells Kenji Oki, Maria W Plonczyski, Milay Luis-Lam, Elise P Gomez-Sanchez, Celso E Gomez-Sanchez GV Montgomery VA Medical Center, Jackson, MS; University of Mississippi, Jackson, MS Primary aldosteronism (PA) is the most common form of secondary hypertension. Approximately half the patients have an aldosterone-producing adenoma (APA). Recently somatic mutations in the selectivity filter of the KCNJ5 gene which codes for the Kir3.4 potassium channel was described in 30-60% of APA patients. Kir3.4 is expressed in the normal human zona glomerulosa and is a G protein-coupled inward rectifying potassium channel that transfers potassium to outside the cell and hyperpolarizes the membrane, thereby opening Calcium channels, leading to the stimulation of aldosterone synthesis. We studied KCNJ5 expression and activity in HAC15 cells derived from H295R cells originating from a human adrenocortical carcinoma. Angiotensin II (A-II) treatment significantly suppressed KCNJ5 mRNA expression (41.2%) and protein (47.3%), as did the calcium ionophore A23187 also suppressed KCNJ5 mRNA levels, and increased aldo synthesis. Naringin, a Kir3.4 activator, significantly abrogated the A-II mediated increase in membrane voltage and aldosterone production was inhibited.[br]Over-expression of the wild type KCNJ5 using a lentiviral vector caused a decrease in membrane voltage, intracellular calcium concentration and aldosterone synthesis under basal and A-II stimulation compared to control cells. mRNA expression of StAR protein and the steroidogenic enzymes HSD3B2, CYP11B1 and CYP11B2 were also significantly suppressed in the KCNJ5 overexpressing cells under basal and A-II stimulation. Knockdown of KCNJ5 with a lentivirus delivering shRNAs for KCNJ5 decreased Kir3.4 protein, but had no effect on aldo synthesis. In conclusion, that A-II mediated aldo biosynthesis is partially mediated by the expression and activity of the potassium channel KCNJ5.[br][br]Sources of Research Support: These studies were supported by grants from the NIH HL27255 and HL105383, and from medical research funds from the Department of Veterans Affairs.[br][br]Nothing to Disclose: KO, MWP, ML-L, EPG-S, CEG-S 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1354 57 349 SAT-246 PO09-01 Saturday 288 2012


289 ENDO12L_SAT-247 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Angiotensin II-Induced Protein Kinase D Activation Partly Mediates Steroidogenic Acute Regulatory Protein Expression and Phosphorylation of Activating Transcription Factor-2 To Underlie Increased Aldosterone Production Lawrence O Olala, Wendy B Bollag Georgia Health Sciences University, Augusta, GA Aldosterone production involves the transport of cholesterol from the outer to the inner mitochondrial membrane where various regulatory enzymes act on this lipid to synthesize the steroid aldosterone. Steroidogenic acute regulatory (StAR) protein mediates this process and is the rate limiting step in steroidogenesis. The serine/threonine protein kinase D (PKD) has been shown to be activated by angiotensin II (AngII) in bovine adrenal glomerulosa cells, underlying increased aldosterone secretion. Previous results from our laboratory obtained from overexpression of various PKD adenoviral mutants, have demonstrated a role for protein kinase C (PKC) and Src family kinases in mediating this process. Most recently, we have shown that overexpression of consitutively active PKD increases StAR mRNA expression. In this study, we also show that PKD promotes the phosphorylation of activating transcription factor-2 (ATF-2), a member of the ATF/cAMP response element (CRE-) binding proteins (CREB) family of leucine zipper proteins, which has been previously shown to bind the StAR proximal promoter thus recruiting coactivator CREB binding protein (CBP) to promote aldosterone secretion.[br]Overexpression of constitutively active serine-738/742[ndash]to-glutamate PKD mutant induced increased ATF-2 phosphorylation and dominant negative acting serine-738/742-to-alanine PKD mutant decreased ATF-2 phosphorylation. Overexpression and infection were verified by western analysis with an antibody recognizing total PKD. In summary, our results demonstrate that AngII activated PKD may induce StAR mRNA expression through increasing the phosphorylation of ATF-2.[br][br]Nothing to Disclose: LOO, WBB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2071 57 350 SAT-247 PO09-01 Saturday 289 2012


290 ENDO12L_SAT-248 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Tannic Acid, an Inhibitor for Renal Angiotensin Type 1 Receptor and Hypertension in Spontaneously Hypertensive Rats Thomas John Thekkumkara, Upendra Gumaste, Russell O Snyder, Vardan Karamyan Texas Technical University Health Sciences Center, Amarillo, TX Dietary intake of polyphenols is associated with lower blood pressure and reduced incidence of cardiovascular disease; however, the mechanisms and targets are not well understood. In a recently published study, we have demonstrated that tannic acid (TA), a natural occurring polyphenol present in a variety of foods (nuts, red wine, tea and coffee), down-regulates the angiotensin type 1 receptor (AT1R) through transcriptional repression. Therefore, in the current study we examined the possible protective effects of TA on blood pressure, as well as AT1R protein and mRNA expression in the kidney of spontaneously hypertensive rats (SHR). All studies were performed in parallel with normotensive Wistar-Kyoto rats (WKY) serving as a control. The SHR and WKY were randomly divided into 2 groups of 6 animals, receiving either vehicle control or TA treatment (5 mg/kg body weight/day-IP). Before treatment, the SHRs and WKYs displayed a baseline mean arterial pressure (MAP) of 147.0[plusmn]4.590 and 106.4[plusmn]6.318 mmHg respectively. At the end of the 4-week study, TA treated SHR showed a significant reduction in MAP (128.1[plusmn]4.750 mmHg) compared to animals treated with vehicle alone (164.6[plusmn]3.096 mmHg). In the normotensive animals with or without TA, there was no detectable reduction in blood pressure. Consistent with the reduction in blood pressure, [3H]Angiotensin II specific binding studies on membrane preparations showed significant reduction (79.50[plusmn]21.78%) in AT1R protein expression in renal cortical tissue. Furthermore, mRNA analysis from renal cortical membrane preparations demonstrated that AT1R expression was significantly down regulated in TA treated animals. Serum concentration of circulating AngII in TA treated SHR were slightly elevated (not significant) compared to all other test groups with no significant change in the renin/prorenin concentrations. The reduction in blood pressure was not accompanied by a significant change in heart/body weight ratio or kidney/body weight ratio in TA treated SHR. These results demonstrate for the first time that long-term therapy with TA can attenuate hypertension in SHR through down-regulation of AT1R expression in the kidney, and this may provide an insight to the mechanism of polyphenolic cardioprotection, an observation previously difficult to explain.[br][br]Sources of Research Support: NIH Grant DK072140.[br][br]Nothing to Disclose: TJT, UG, ROS, VK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2291 57 351 SAT-248 PO09-01 Saturday 290 2012


291 ENDO12L_SAT-249 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Promoter-Specific Consequences of the N/C-Interaction in the Mineralocorticoid Receptor Peter J Fuller, Yizhou Yao, David Pearce Prince Henry[apos]s Institute of Medical Research, Clayton, Australia; University of California San Francisco, San Francisco, CA The mineralocorticoid receptor (MR) is unique in responding to two physiological ligands, aldosterone and cortisol. In epithelial tissues aldosterone selectivity is determined by the activity of 11[beta]HSD2. In other tissues, cortisol is the primary ligand; it may act as an antagonist. We have previously characterised an interaction between the N-terminal domain and the C-terminal, ligand-binding domain (LBD) of the human MR (N/C-interaction) which occurs in response to aldosterone but is antagonised by cortisol (Pippal, et al Mol Endocrinol 23: 1360, 2009). Given the equivalence of aldosterone and cortisol with a full-length MR in a simple transactivation assay, it would seem likely that the N/C-interaction is not obligatory for MR action but it may have a role in gene or tissue-specific signalling. The structural basis of this ligand-discriminant interaction has been elusive. We have recently found that two amino acids, serine 936 and glutamine 940 in helix 11 of the LBD have a role in the N/C-interaction. Mutation of each alone, S936R and E940R, decreased the aldosterone-induced interaction in the mammalian 2-hybrid assay; the double mutation (DM) eliminated the interaction. In a transactivation assay using CV-1 cells with an MMTV-LUC reporter, S936R, E940R or DM had no effect on the response. DM thus dissociates the MR N/C-interaction from the transactivation response. The ligand and promoter-dependence of this dissociation was examined. The responses of MMTV-LUC were not different with cortisol nor in the presence of an MR ligand-specific coactivator, tesmin. The response to aldosterone was compromised for DM when compared to MR at the minimal promoters TAT1 and TAT3, with a greater relative impact on the TAT3 promoter. There was no right-shift of the aldosterone dose response curve with DM indicating that the effect was not a consequence of a change in ligand-binding affinity. Transactivation at two physiological promoters GILZ and Cnksr3 was dichotomous. Whilst the response with Cnksr-LUC paralleled that with MMTV-LUC, the response with GILZ-LUC for DM was [lt] 50% that of MR. The molecular basis of this promoter specific differential response is currently being explored. It suggests that the N/C-interaction maybe important at multimer response elements, perhaps through an intermolecular interaction. These studies provide a unique insight into the contribution of the N/C-interaction to providing diversity of responses at the MR.[br][br]Sources of Research Support: National Health and Medical Research Council of Australia.[br][br]Nothing to Disclose: PJF, YY, DP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 508 57 352 SAT-249 PO09-01 Saturday 291 2012


292 ENDO12L_SAT-250 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Mutations in KCNJ5 Alter the Transcriptome of Aldosterone-Producing Adenomas and Human Adrenocortical Cells Namita G Hattangady, Silvia Monticone, Franco Mantero, Beatrice Rubin, Maria Verena Cicala, Raffaele Pezzani, Richard J Auchus, Tracy A Williams, William E Rainey Georgia Health Sciences University, Augusta, GA; University of Padova, Padova, Italy; University of Michigan Medical School, Ann Arbor, MI; University of Torino, Torino, Italy [bold]Introduction[/bold]: Adrenal aldosterone production and expression of the enzyme aldosterone synthase (CYP11B2) expression are normally controlled by the renin-angiotensin system. Up to 10% of hypertensive patients, however, exhibit elevated aldosterone production and CYP11B2 expression despite suppressed or low circulating renin, a condition termed primary aldosteronism (PA). PA has two primary causes: idiopathic hyperaldosteronism (IHA) and unilateral adrenal aldosterone producing adenoma (APA). Recently, somatic point mutations in the potassium channel, KCNJ5, were found in greater than one third of APA and implicated in increased aldosterone production via depolarization of the adenoma cells. [bold]Objective[/bold]: To define the effect of KCNJ5 mutations on gene expression and aldosterone production using APA tissue and a human adrenocortical cell line (HAC15). [bold]Methods[/bold]: APA (n=19) were obtained from two different centers (Padua, Italy and Dallas, USA) and categorized based on sequencing as APA expressing wild type or mutant KCNJ5. Microarray analysis was used to compare gene expression profiles in APA with and without KCNJ5 mutations, and in HAC15 cells over-expressing mutant and wild-type KCNJ5. Quantitative real time RT-PCR (qPCR) validated a set of differentially expressed genes. [bold]Results[/bold]: Microarray comparison was made between 11 APA with mutations in KCNJ5 (7 G151R and 4 L168R) and 8 tumors without KCNJ5 mutation. Microarray comparison of APA with mutant KCNJ5 (versus APA with wildtype KCNJ5) indicated a significant up-regulation in 31 genes and down-regulation in 29 genes by [gt]2.0 fold (p[lt]0.05). Moreover, aldosterone synthase (CYP11B2) was the highest up-regulated transcript in APA with mutant KCNJ5 (3 fold). qPCR analysis on a larger APA subset also indicated a 6 fold increase in CYP11B2 expression in APA with mutated KCNJ5. Supporting these observations, over-expression of the G151R and L168R KCNJ5 mutations in HAC15 cells increased aldosterone production, elevated CYP11B2 expression (9 fold) and altered 36 genes by [gt]2.5-fold (p[lt]0.05). qPCR confirmed increased expression of CYP11B2, its essential transcription factor nuclear receptor subfamily 4, group A, member 2 (NR4A2) and nuclear receptor subfamily 4, group A, member 3 (NR4A3). [bold]Conclusions[/bold]: Our data suggest that KCNJ5 mutations increase expression of CYP11B2 and its transcriptional regulators, NR4A2 and NR4A3, thus leading to a primary aldosteronism phenotype exhibiting aldosterone over-production.[br][br]Disclosures: RJA: Investigator, Jansen Pharmaceuticals. Nothing to Disclose: NGH, SM, FM, BR, MVC, RP, TAW, WER 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2227 57 353 SAT-250 PO09-01 Saturday 292 2012


293 ENDO12L_SAT-251 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Human 11[beta]-Hydroxysteroid-Dehydrogenase Type 2 (11BHSD2) Knockdown Affects Mineralocorticoid Response Genes in the Endothelial Cell Line EaHy926 Cristian A Carvajal, Alejandra Tapia, Cristobal A Fuentes, Carolina DP Valdivia, Carlos F Lagos, Fidel Allende, Sandra Solari, Carmen Campino, Rene F Baudrand, Alejandra C Martinez, Gareth I Owen, Carlos E Fardella Pontificia Universidad Catolica de Chile, Santiago, Chile; Pontificia Universidad Catolica de Chile, Santiago, Chile; Pontificia Universidad Catolica de Chile, Santiago, Chile; Pontificia Universidad Catolica de Chile, Santiago, Chile High blood pressure affects 26% of the world population. We have reported that 15% essential hypertensives may suffer from impairment of the enzyme 11beta-hydroxysteroid-dehydrogenase type 2 (11BHSD2), which inactivates cortisol to cortisone, thus avoiding mineralocorticoid receptor (MR) activation. [bold]Aim[/bold]: In EaHy926 cells, evaluate via HSD11B2-knockdown the expression of genes associated with mineralocorticoid activity and inflammatory response. [bold]Methods[/bold]: The human vascular endothelial cell line EaHy926 (ATCC, CRL-2922) was cultured at 70-80% confluence in IMDM/10% FBS charcoal-treated (steroid free), and subsequently transiently transfected with stealth siRNA-HSD2 to knockdown (KD) the human HSD11B2 RNA, prior to 50 nM cortisol (F) challenge for 18 hrs. HSD11B2, MR and 18S RNA were quantified by Taqman RT-qPCR. The expression of genes associated with MR activation (alpha-ENAC, beta-ENaC, WNK1, NEdd4-2, and Na/K-ATPse) and inflammatory response (TNFa, IL-10, TGFB) were quantified by Sybr-green qPCR. The results are expressed as X[plusmn]SD in arbitrary units (AU) respect to control. Statistical comparisons were performed by either Mann-Whitney or t-test. [bold]Results[/bold]: EaHy926 cells showed an increase of HSD11B2-RNA expression when treated with F (1.02[plusmn]0.24 vs. 1.32[plusmn]0.15, p 0.05). EaHy926 transiently transfected with HSD11B2-KD reduce HSD11B2-RNA with either treatment with vehicle (V) (0.10[plusmn]0.15, p 0.001) or F (0.42[plusmn]0.16, p 0.01), respectively. The RNA expression in HSD11B2-KD cells treated with F showed a decrease in MR (1.31[plusmn]0.15 vs. 0.66[plusmn]0.16, p 0.01), beta-ENAC (1.40[plusmn]0.15 vs. 0.59[plusmn]0.16, p 0.01), WNK1 (2.00[plusmn]0.45 vs. 0.39[plusmn]0.36, p 0.001) and NEdd4-2 (0.81[plusmn]0.19 vs. 0.41[plusmn]0.12, p 0.05). We did not observe changes in the expression alpha-ENAC (0.95[plusmn]0.22 vs. 1.03[plusmn]0.30, pNS) or Na/K-ATPse (0.78[plusmn]0.35 vs. 1.16[plusmn]0.56, pNS). Assayed cytokines displayed no significant changes in expression in either control or F treated EaHy926 HSD11B2-KD cells. [bold]Conclusion[/bold]: In EaHy926 cells, siRNA-HSD2 decreased the 11BHSD2-RNA expression in both basal and stimulated (F, 50nM) conditions, which concomitantly associate a reduced expression of MR, beta-ENaC, WNK1 and Nedd4-2, suggesting these genes -downstream of MR- are also affected by HSD11B2-KD. Inflammatory response genes analyzed were not affected by F or HSD11B2-KD. Further studies using both KD and pharmacological inhibitors are necessary to reinforce these results, and clear the role of 11BHSD2 deficiency in mineralocorticoid hypertension.[br][br]Sources of Research Support: FONDEF D08i1087, FONDECYT 1100356 [amp] IMII P09/016-F Grants supported this work. CFL and CAC are PhD fellows from Comision Nacional y Cientifica de Chile (CONICYT).[br][br]Nothing to Disclose: CAC, AT, CAF, CDPV, CFL, FA, SS, CC, RFB, ACM, GIO, CEF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 350 57 354 SAT-251 PO09-01 Saturday 293 2012


294 ENDO12L_SAT-252 POSTER SESSION: Endocrine Hypertension: Renin-Angiotensin-Aldosterone System (1:30 PM-3:30 PM) Effects of BMP-6 on Aldosterone Production by Rat Adrenal Gland [italic]In Vivo[/italic] Tomoko Miyoshi, Fumio Otsuka, Yoshinori Matsumoto, Kenichi Inagaki, Naoko Tsukamoto, Eri Nakamura, Mariko Takano-Narazaki, Kanako Ogura-Ochi, Masaya Takeda, Hirofumi Makino Okayama University Graduate School, Okayama, Japan Aldosterone is synthesized in the zona glomerulosa of the adrenal cortex. We previously reported the presence of a functional BMP/activin system including BMP-6 and BMP type-1 and type-2 receptors in human adrenocortical cells. BMP-6 contributes to Ang II-induced aldosterone production by activating Smad1/5/8 through binding ALK-2 and/or ALK-3 in combination with ActRII and by enhancing Ang II-induced MAPK activation, implying that this machinery of aldosterone regulation may be involved in the process of the aldosterone escape phenomenon. As for an in vivo study on BMP-6, no particular phenotype regarding endocrine abnormality has been documented in BMP-6 null mice with the exception of delayed skeletogenesis in the sternum. In the present study, we attempted to elucidate the physiological role of BMP-6 in regulation of aldosterone in vivo, in a harmless way by utilizing rats treated with immunization against BMP-6. Three-week-old male rats were actively immunized with rat mature BMP-6 antigen conjugated with keyhole limpet hemocyanin (KLH). The immunization treatment had no effect on bilateral adrenal weight or its ratio to body weight. Urinary aldosterone excretion was time-dependently increased during the 8-week observation period in the control group. Of note, the level of urinary aldosterone excretion in BMP-6-KLH-immunized rats was significantly reduced compared to that in the control group, suggesting that endogenous BMP-6 contributes to the induction of aldosterone production in vivo. Moreover, the level of urinary aldosterone/creatinine after 8-week treatment was significantly lowered by treatment with BMP-6-KLH. In contrast, with chronic Ang II treatment, urinary aldosterone and creatinine-corrected values at 8 weeks were not significantly different between the two groups, suggesting that the effects of BMP-6-KLH were impaired under the condition of chronic treatment with Ang II. The expression levels of CYP11B2 mRNA were significantly decreased in adrenal tissues isolated from BMP-6-KLH-immunized rats after 8-week treatment. Furthermore, the ratio of plasma aldosterone level to corticosterone was significantly decreased by immunization with BMP-6-KLH. Collectively, the results indicate that endogenous BMP-6 is functionally linked to aldosterone synthesis by the zona glomerulosa in the adrenal cortex in vivo.[br][br]Nothing to Disclose: TM, FO, YM, KI, NT, EN, MT-N, KO-O, MT, HM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1492 57 355 SAT-252 PO09-01 Saturday 294 2012


295 ENDO12L_SAT-254 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) SRC-3 Is a Critical Mediator in the Development of Castration-Resistant Prostate Cancer Jean Ching-Yi Tien, Zhaoliang Liu, Gao Li, Fen Wang, Jianming Xu Texas A[amp]M Health Science Center, Houston, TX; Baylor College of Medicine, Houston, TX; MD Anderson Cancer Center, Houston, TX Androgen ablation therapy is standard treatment for advanced prostate cancer. While tumors initially respond to androgen deprivation, they almost always recur in an androgen-independent phenotype termed castration-resistant prostate cancer (CRPC). Castration-resistant tumors are incurable with current therapy regimens and account for most prostate cancer mortality. Better understanding of their biology is therefore critical for devising effective treatment options. Steroid Receptor Coactivator 3 (SRC-3) is a nuclear receptor coactivator that promotes growth of endocrine tissues. It enhances proliferation of prostate cancer cell lines and is highly expressed in advanced human prostate tumors. Despite this, the role of SRC-3 in CRPC is not well studied. PTEN is a tumor suppressor gene mutated in most human prostate cancers. Mice harboring PTEN deletion in prostatic epithelium develop cancer that arises from progenitors in the luminal epithelial and basal compartments. Tumors histologically mimic advanced human disease and are castration-resistant. [bold]We hypothesized that, in prostate tumors caused by PTEN deletion, SRC-3 is a critical mediator in accelerating the development of CRPC. [/bold]To test this hypothesis, we generated mice in which floxed [italic]PTEN[/italic] and [italic]SRC-3[/italic] genes were concomitantly deleted in prostate epithelial cells via a Cre recombinase driven by the Probasin promoter. We first compared tumor mass, histology and biomarkers in these PTEN[sup]f/f[/sup]/SRC-3[sup]f/f[/sup]/ARR2PBiCre (pten3cko) mice vs. PTEN[sup]f/f[/sup]/ARR2PBiCre (ptencko) mice. We found that tumor histology and cellular composition were not markedly different between the groups at 12wks. We then castrated mice of both genotypes at 9-wks-old and harvested tumors at 12wks. Interestingly, castrated ptencko mice had significantly worsened tumor aggressiveness--increased proliferation index, reduced differentiation (increased p63; decreased K5 and K8) and increased reactive stroma (double staining of SMA and vimentin)--versus their non-castrated counterparts. In addition to significantly reducing tumor size, SRC-3 ablation reversed all changes comprising the aggressive phenotype seen in the post-castration tumors. Interestingly, SRC-3 deletion decreased mTOR signaling. Together, these data show castration worsens phenotype of tumors caused by PTEN deletion and that SRC-3 promotes this process by enhancing epithelial cell proliferation trophic effect onstroma. SRC-3 may serve as a potential target for CRPC therapy.[br][br]Nothing to Disclose: JC-YT, ZL, GL, FW, JX 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1299 58 356 SAT-254 PO42-01 Saturday 295 2012


296 ENDO12L_SAT-255 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) The Role of Progesterone Receptor as a Suppressor of Epithelial-Mesenchymal Transition in Breast Cancer Adriana P Visbal, Viroj Boonyaratanakornkit, Dean P Edwards Baylor College of Medicine, Houston, TX; Chulalongkorn University, Bangkok, Thailand Progesterone (P4) action via progesterone receptor (PR) results in increased proliferation of breast epithelial cells and several lines of evidence suggest that P4 exposure is a risk factor for developing breast cancer. Despite the mitogenic activities of P4, the presence of PR in breast cancer is associated with a better prognosis, a better response to endocrine therapy, and a more differentiated phenotype suggesting that PR may have suppressor activity during cancer progression. In T47DY breast cancer cells, a PR negative variant of T47D, loss of PR is associated with a change in cell morphology to a more mesenchymal-like shape, increased cell motility and up-regulation of several epithelial-messenchymal transition (EMT) associated genes. Re-expression of PR in this cell line reversed these EMT-like phenotypes. shRNA-mediated knockdown of PR in parental T47D cells resulted in up-regulated gene expression of vimentin and slug, two genes known to play a crucial role in the EMT process. EMT is characterized by morphological changes in cytoskleletal architecture and cell polarity that lead to cell invasiveness. In order to investigate the role of PR in these processes, we developed a 3D acini Matrigel culture system with non-transformed MCF10A human breast epithelial cells in which PR expression is regulated from a tetracycline promoter by doxycycline (Dox). We examined the effect of PR expression, with and without P4, on EMT induced by transforming growth factor [beta] (Tgf[beta]). Morphological changes characteristic of EMT in 3D cultures were analyzed by immunostaining for epithelial cell polarity markers and by fluorescent labeling of the actin cytoskeleton. Upon Tgf[beta] exposure, acini become smaller, amorphous, and exhibit actin cytoskeleton protrusions. Additionally, changes in the phosphorylation of the ezrin/radixin/moesin (ERM) family of actin binding proteins are also observed. In Dox-treated acini that express PR, these morphological changes are less pronounced and phospho-ERM levels appear to be restored to those of non-Tgf[beta] treated acini. The supression of these Tgf[beta]-induced phenotypes is most predominant in acini exhibiting the highest levels of PR expression. These data support the concept that PR can suppress EMT during breast cancer progression. On going studies to examine the ability of PR expression to suppress EMT in highly metastatic breast cancer cell lines [italic]in vitro[/italic] and metastatic potential with [italic]in vivo[/italic] mouse models will be reported.[br][br]Nothing to Disclose: APV, VB, DPE 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1728 58 357 SAT-255 PO42-01 Saturday 296 2012


297 ENDO12L_SAT-256 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Plasma Steroid Concentrations in Male Rhesus Monkeys Following Treatment with the P450c17 (CYP17) Inhibitors VT-464 and Abiraterone Acetate: A Comparison to Human 17,20-Lyase (lyase) and Combined Lyase/17[alpha]-Hydroxylase (hydroxylase) Deficiencies David H Abbott, Joel R Eisner, Ian M Bird, Stephen W Rafferty, William R Moore, Robert J Schotzinger University of Wisconsin, Madison, WI; Viamet Pharmaceuticals, Inc, Morrisville, NC; University of Wisconsin, Madison, WI CYP17 is a single protein with two distinct P-450-dependent enzyme activities, hydroxylase and lyase. Reduction of extra-gonadal androgen production through CYP17 inhibition is a validated paradigm for treatment of castration-resistant prostate cancer and may prove beneficial for the treatment of disorders of androgen excess such as polycystic ovary syndrome. Individuals with mutations within the CYP17 coding region are either lyase-deficient or are both lyase and hydroxylase deficient. Both groups have decreased plasma testosterone (T) concentrations, but only those that are lyase/hydroxylase -deficient have accompanying increased progesterone (P4) and decreased cortisol (F) concentrations [1,2]. The objective of the current study was to compare the endogenous steroid hormone responses to the CYP17 inhibitors VT-464 and abiraterone acetate (AA) in male rhesus monkeys to the endogenous steroid hormone responses to genetically-determined lyase or lyase/hydroxylase deficiencies in humans. Plasma T, P4, and F concentrations were determined using LC/MS/MS methods after a single subcutaneous dose (12.5 mg/kg) of VT-464, AA, or vehicle. The % change from vehicle for VT-464- and AA-treated animals was determined at each time-point (0-24 h). The single values reported below represent the time-point as which the greatest reduction in T occurred (% of vehicle). The % change for the human deficiencies was calculated from the median normal reference range values reported. A comparison of steroid hormone concentrations following VT-464 or AA treatment in primates (% of vehicle) with those from lyase-deficient or LHD humans (% of normal) are: T [9% (VT-464), 3% (AA), 30% (lyase-deficient), and 4% (lyase/hydroxylase-deficient)]; P4 [100% (VT-464), 1,494% (AA), 212% (lyase-deficient), and 1,000% (lyase/hydroxylase-deficient)]; F [82% (VT-464), 9% (AA), 103% (lyase-deficient), and 26% (lyase/hydroxylase-deficient)]. Similar to lyase- and lyase/hydroxylase-deficient individuals, T concentrations were reduced in both VT-464- and AA-treated primates. However, the effects of VT-464 and AA treatments on P4 and F segregated with isolated lyase and combined lyase/hydroxylase deficiencies, respectively. The current study demonstrates that selective CYP17 lyase inhibition effectively suppresses T concentrations without significantly perturbing F or P4 concentrations.[br][br][1] Martin et al, J Clin Endocrinol Metab 88: 5739-5746, 2003. [2] Kok et al, J Clin Endocrinol Metab 95: 994-999, 2012.[br][br]Disclosures: DHA: Ad Hoc Consultant, Viamet Pharmaceuticals, Inc.; Recipient Award, Viamet Pharmaceuticals, Inc. JRE: Employee, Viamet Pharmaceuticals, Inc. IMB: Consultant, Viamet Pharmaceuticals, Inc. SWR: Employee, Viamet Pharmaceuticals, Inc. WRM: Employee, Viamet Pharmaceuticals, Inc. RJS: Employee, Viamet Pharmaceuticals, Inc. 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1622 58 358 SAT-256 PO42-01 Saturday 297 2012


298 ENDO12L_SAT-257 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Mammalian Target of Rapamycin Activation in Estrogen Receptor [alpha] Positive Human Breast Tumor Is a Predictor for Better Treatment Outcome Anuraag Shrivastav, Srinivas Seekallu, Zoann Zoann Nugent, Leigh C Murphy University of Manitoba and CancerCare Manitoba, Winnipeg, Canada; University of Winnipeg, Winnipeg, Canada; University of Manitoba and CancerCare Manitoba, Winnipeg, Canada Expression of estrogen receptor (ER-[alpha]) is an important determinant for employing hormonal therapy for breast cancer. Earlier we provided a phosphorylation score, called P7 score, which represents the presence of up to seven phosphorylation-ER[alpha] sites detected in a tumor. On multivariate analysis the P7score is significantly associated with over-all survival (OS) from breast cancer death and relapse free survival (RFS) in ER-[alpha] positive breast cancer patients who were later treated with tamoxifen. The P7score represents the balance of those phospho-ER[alpha] sites associate with good versus poor clinical outcome (1) Mammalian target of rapamycin (mTOR) is a central hub of signaling pathways impacting cell growth, transformation, proliferation and protein synthesis. Since mTOR activation is implicated in resistant to endocrine therapy in breast cancer we were interested in investigating whether mTORC1 activation, as measured by its phosphorylation on S2448 residue (p-mTOR), was associated with OS and RFS in the same patient cohort used above. Contrary to earlier reports we observed that p-mTOR expression, measured by IHC, was negatively associated with size and nodal status (Spearman correlation r = -0.12, P= 0.026, n=337; r = -0.15, P = 0.0068, n=331, respectively). Furthermore, p-S2448 mTOR expression was found to be positively correlated with p-S118-ERa (r = 0.268, n= 308, P [lt] 0.0001), p-S167-ERa (r = 0.205, n= 325, P = 0.0002) and p-S282-ERa (r = 0.188, n= 304, P=0.001) but negatively correlated with p-T311-ERa (r = -0.125, n=307, P = 0.028). Consistent with these findings we also observed that p-mTOR was negatively associated with P7 score (r = -0.23, n = 284, P [lt] 0.0001) and phosphorylation of mTOR at S2448 residue was significantly associated with OS (HR = 0.61, P= 0.028, 95% CI 0.39-0.95, n = 337) and RFS (HR = 0.58, P = 0.0032, 95%CI 0.41-0.83, n = 337) in a cohort of ER-[alpha] positive primary breast tumor cases wherein patients were later treated with tamoxifen. However, in contrast to the P7-score, p-mTOR relationship to clinical outcome did not remain significant on multivariate analysis. In summary, we report here for the first time that activated mTORC1 is a marker of an intact estrogen dependent signaling pathway in breast cancer and therefore likely to be predictive of better response to the endocrine therapies.[br][br](1) Skliris et al., Endocr. Relat. Cancer2010,17:589.[br][br]Sources of Research Support: This work was supported by the Canadian Institute of Health Research (CIHR), the CancerCare Manitoba Foundation (CCMF) and the Canadian Breast Cancer Foundation (CBCF). This study was also supported by the Manitoba Breast Tumor Bank, Winnipeg, Manitoba, Canada funded in part by CCMF and CIHR.[br][br]Nothing to Disclose: AS, SS, ZZN, LCM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2295 58 359 SAT-257 PO42-01 Saturday 298 2012


299 ENDO12L_SAT-258 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Androgenic Regulation of the Anti-Apoptotic Bcl-2 Family Member Mcl-1 in Prostate Cancer Cell Lines Frederic R Santer, Su Jung Oh, Zoran Culig Innsbruck Medical University, Innsbruck, Austria Anti-apoptotic proteins of the Bcl-2 family are known to confer resistance against a variety of therapeutic agents. Among them is induced myeloid leukemia cell differentiation protein (Mcl-1), a highly regulated protein at both transcriptional and post-translational levels by a number of signaling pathways. Overexpression of Mcl-1 has been reported in biopsies of prostate cancer patients with high Gleason grade. We have previously shown that Mcl-1 mediates the anti-apoptotic effect of the Interleukin-6 autocrine loop in a cellular model of advanced prostate cancer. Currently, novel anti-cancer approaches that aim to inhibit Mcl-1 are being developed. In this study we have analyzed the influence of androgens and Androgen Receptor (AR) signaling on Mcl-1 protein expression. Western Blot experiments with androgen-sensitive prostate cancer cell lines LNCaP and VCaP showed a strong and consistent upregulation of Mcl-1 protein expression levels under androgen-depleted conditions, i.e. in steroid-free medium. Addition of 0.1-10 nM of the synthetic androgen R1881 decreased Mcl-1 expression in a concentration-dependent manner and the maximal effect was observed after 24-48h incubation time. Androgenic regulation of Mcl-1 was dependent on the presence of functional AR since Mcl-1 downregulation could not be observed in cells co-treated with R1881 and antiandrogens bicalutamide or hydroxyflutamide or after AR down-regulation by siRNA. Interestingly, Mcl-1 mRNA expression was unchanged after treatment with R1881 as measured with qPCR leading to the hypothesis that the decrease of Mcl-1 is a post-translational mechanism. This was corroborated by addition of the proteasome inhibitor MG132 that stabilized Mcl-1 protein expression levels. We therefore hypothesize that AR could regulate the expression of an E3-Ligase able to polyubiquitinate Mcl-1 and target it to the proteasome. The finding of androgenic regulation of the anti-apoptotic protein Mcl-1 may explain its high expression in specimens obtained from high Gleason grade prostate cancer patients. Androgen ablation might lead to upregulated Mcl-1 levels thus contributing to the development of resistances against first- and second-line therapies. Therefore, anti-Mcl-1 therapies might be considered to improve the efficacy of androgen ablation in patients with prostate cancer.[br][br]Sources of Research Support: Austrian National Bank to Z.C.[br][br]Disclosures: ZC: Principal Investigator, Johnson & Johnson. Nothing to Disclose: FRS, SJO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 795 58 360 SAT-258 PO42-01 Saturday 299 2012


300 ENDO12L_SAT-259 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Reprogramming of Estrogen Receptor Signaling by the Serine Threonine Kinase AKT in Breast Cancer Cells Harikrishna Nakshatri, Guohua Wang, Yunlong Liu, Sunil Badve, Poornima Bhat-Nakshatri Indiana University School of Medicine, Indianapolis, IN; Indiana University School of Medicine, Indianapolis, IN; Indiana University School of Medicine, Indianapolis, IN Activating mutation of PI3Kinase is more common in luminal type A subtype of breast cancers. A transcription factor network comprising estrogen receptor alpha (ER), FOXA1, and GATA-3 confers estrogen (E2) dependent growth properties to luminal type A breast cancer. FOXA1 and GATA-3 serve as [ldquo]pioneer factors[rdquo] that influence the ER binding pattern to the genome. Previous studies by others and us have shown that PI3 kinase and its downstream kinase AKT are integral parts of ER-E2 signaling in breast cancer. AKT-mediated changes in E2 signaling are associated with altered E2-dependent ER-binding suggesting the ability of AKT to reprogram ER binding to the genome. We designed this study using MCF-7 cells and MCF-7 cells overexpressing constitutively active AKT (CA-AKT) as a model system to test the following: 1) the effect of AKT on ER binding to genes in Oncotype DX and MapQuantDxTM gene signatures, which are used as prognostic tests for ER-positive breast cancer; 2) the effect of AKT on E2-regulated expression of genes in these signatures and 3) transcription factors that may cooperate with ER and AKT to alter the expression of genes in MapQuantDxTM. E2 increased the expression of five among 16 genes of the Oncotype DX; AKT further increased expression of one of them. AKT had no effect on five genes that were repressed by E2. Among 97 genes of MapQuant DXTM, 42 and 50 genes were E2 inducible in MCF-7 and MCF-7CA-AKT cells, respectively. Interestingly, only five and nine genes in MCF-7 and MCF-7CA-AKT cells, respectively, contained ER binding sites. These results suggest that AKT alters secondary E2-response pathway to modulate the expression of genes in the MapQuantDXTM. Transcription factor motif search analysis revealed MapQuantDXTM signature genes commonly expressed in both cell types were enriched for E2F, Rb:E2F1:DP1, NF-1, and TBX5 transcription factor binding sites. Myc:Max and NRF2 binding sites were enriched in genes that are uniquely E2-regulated in MCF-7 cells, whereas STAT3, STAT5A, STAT6, LBP1, E2F2:DP2, E2F4:DP2, and NF-kappaB binding sites were enriched in genes uniquely E2-regulated in MCF-7CA-AKT cells. The basal and E2-regulated expression of E2F family transcription factors was different in MCF-7 and MCF-7CA-AKT cells. From these results, we conclude that PI3 Kinase and its downstream target AKT alter E2 secondary response, which ultimately affects the expression of genes that constitute prognostic signatures of ER-positive breast cancer.[br][br]Nothing to Disclose: HN, GW, YL, SB, PB-N 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2069 58 361 SAT-259 PO42-01 Saturday 300 2012


301 ENDO12L_SAT-260 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Effects of Aromatase Inhibitor Therapy on Alteration of Enzymes Involved in Intratumoral Estrogen Production in Lung Carcinoma Cells Mohit Kumar Verma, Yasuhiro Miki, Hironobu Sasano Tohoku University School of Medicine, Sendai-shi, Japan [bold]Background[/bold]: Recent studies demonstrated estrogens were synthesized [italic]in situ[/italic] in non-small cell lung carcinomas (NSCLC) through aromatase. It was further demonstrated that aromatase inhibitor (AI) suppressed the NSCLC growth. In breast cancer patients, AI therapy may lead to emergence of alternative intratumoral estrogen production pathways. Similar emergence of alternative intratumoral estrogen production pathway may occur in NSCLC patients following AI therapy. Therefore, the aim of the present study was to elucidate the effects of letrozole (AI) upon the enzymes involved in intratumoral estrogen production in NSCLCs.[br][bold]Methods[/bold]: We employed NSCLC cells i.e. A549 [amp] LK87 and evaluated expression of enzymes including aromatase, steroid sulfatase (STS) and 17-[beta] hydroxysteroid dehydrogenases (17-[beta]HSD type I [amp] II) following letrozole treatment using both q-RT-PCR and immunoblotting. In addition, archival materials form 104 NSCLC patients were immunohistochemically evaluated using anti-17-[beta]HSD type I [amp] II antibodies.[br][bold]Results[/bold]: Letrozole treatment significantly decreased the cell proliferation of A549 and LK87 cells (p=[lt].0001). Letrozole treatment also suppressed the cell migration in A549 cells (p=[lt].0001). In addition, letrozole treatment induced both mRNA and protein levels of both 17-[beta]HSD type I and II enzymes in both cell lines (p=[lt].0001). Letrozole treatment also decreased aromatase protein levels in both the cell lines but no differences were detected in STS expression. NSCLC patients with a higher immunoreactivity of 17-[beta]HSD type II were associated with significantly larger tumor diameter (p=.002), higher tumor stage (p=.003) and tended to be associated with the presence of lymph node metastasis (p=.057). In addition, the patients with a higher expression of 17-[beta]HSD type I tended to be associated with higher tumor stage (p=.053) and the presence of lymph node metastasis (p=.089) but did not reach statistical significance. A subsequent multivariate regression analysis revealed a higher 17-[beta]HSD type I status (HR= 2.58, p=0.017) as an independent prognostic factor.[br][bold]Conclusion[/bold]: Our results suggest that 17-[beta]HSD pathway may be considered as an important prognostic factor in NSCLC patients and its modulation following AI therapy may represent a potent marker for response to aromatase inhibitor therapy in NSCLC patients. Therefore, a combination therapy with AI and 17-[beta]HSD inhibitor may further improve the clinical response in these hormone responsive NSCLC patients.[br][br]Nothing to Disclose: MKV, YM, HS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 648 58 362 SAT-260 PO42-01 Saturday 301 2012


302 ENDO12L_SAT-261 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Co-Expression of Androgen Receptor and 5[alpha]R1 in Triple-Negative Breast Cancer (TNBC) Is Associated with Lower Rates of Tumor Cell Proliferation Keely May McNamara, Yasuhiro Miki, Niramol Chanplakorn, Sansanee Wongwaisayawan, Pimpin Incharoen, Yasuhiro Nakamura, Takashi Suzuki, Minoru Miyashita, Kentaro Tamaki, Takonori Ishida, Noriaki Ohuchi, Hironobu Sasano Tohoku University School of Medicine, Sendai, Japan; Ramathibodi Hospital, Bangkok, Thailand Background: Triple negative breast cancer (TNBC) is a subset of breast cancer defined by the absence of estrogen/progesterone receptors (ER/PR) and HER2 expression in carcinoma cells. The management of TNBC patients poses clinical problems due to ineffective endocrine and Trastuzumab treatment leading to the exploration of potential new therapeutic targets including androgens.[br]Methods: Following the IRB approval of both institutions, we examined two TNBC cohorts, one from Tohoku University Hospital Japan (n= 87), and the other from Ramathibodi Hospital, Bangkok, Thailand (n=121). ER/PR and HER2 were evaluated based upon ASCO/CAP guideline for the confirmation of triple negative status of the cases. Tissue sections were prepared from 10% formalin-fixed and paraffin embedded blocks and androgen receptor (AR), 5 alpha reductase type one (5[alpha]R1) and 17 beta hydroxysteroid dehydrogenase type five (17[beta]HSD5) were evaluated via IHC. Nuclear AR immunoreactivity was quantified using H score. Immunoreactivity of the androgenic enzymes was evaluated using a semi-quantitative method (0: no, 1: 1-50% and 2: 51-100% positive carcinoma cells).[br]Results: Increased intratumoral 5[alpha]R1 and/or 17[beta]HSD was associated with a higher AR score in both Thai and Tohoku TNBC cohorts (n=208) reaching significance (p=0.007) with 5[alpha]R1 but not with 17[beta]HSD. To examine the effect of this interaction on tumor biology we sub-classified the Tohoku TNBC patients into 4 groups (AR high/5[alpha]R1 high, AR high/5[alpha]R1 low, AR low/5[alpha]R1 high, AR low/5[alpha]R1 low). 5[alpha]R1 high was defined as samples having greater than 50% immunoreactivity. AR high was defined by greater than 10% of cells being immunoreactive. The Ki67 labelling index in AR high/5[alpha]R1 high TNBCs was significantly lower (p=0.005) than that of AR low/5[alpha]R1 high or AR low/5[alpha]R1 low TNBCs (24% vs. 62 and 49% respectively).[br]Conclusions: AR status of carcinoma cells was significantly associated with increased levels of 5[alpha]R1 expression in TNBCs. These results suggests that AR positive TNBC cases actively synthesize androgen in situ from circulating adrenal androgen precursors and AR detected in these cases may be considered functional. The co-expression of both androgen receptor and androgen producing enzymes results in decreased carcinoma cell proliferation in TNBCs suggesting that therapeutic approaches that stimulate androgenic signalling could be effective in AR positive triple negative breast cancer patients through decreasing carcinoma cell proliferation.[br][br]Nothing to Disclose: KMM, YM, NC, SW, PI, YN, TS, MM, KT, TI, NO, HS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1381 58 363 SAT-261 PO42-01 Saturday 302 2012


303 ENDO12L_SAT-262 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Prostate Cancer Cells Are More Responsive to Testosterone or Prolactin Induction of Carboxypeptidase-D Than Benign Prostate Cells Lynn Natalie Thomas, Catherine Kar-Ling Too Dalhousie University, Halifax, Canada; Dalhousie University, Halifax, Canada [bold]Introduction:[/bold] Carboxypeptidase-D (CPD) is a membrane-bound metalloproteinase that cleaves C-terminal arginine, which is the substrate of nitric oxide synthase for the production of nitric oxide (NO). NO regulates many physiological processes, including tumor progression and angiogenesis. Previously, we reported that CPD and NO levels were upregulated by testosterone (T) or prolactin (PRL) in prostate cancer (pCa) cells, which enhanced cell survival (1). In the current study, T/PRL regulation of CPD expression was further characterized. CPD and NO levels in benign and malignant prostate cells/tissues were compared to determine their roles in pCa development.[br][bold]Methods:[/bold] Quantitative RT-PCR (QPCR) was used to measure CPD mRNA levels in human prostate cell lines. Immunohistochemistry (IHC) and tissue microarray analysis (TMA) were used to compare CPD staining in human prostate specimens. NO production was measured using 4,5-diaminofluorescein diacetate (DAF-2DA) assay.[br][bold]Results:[/bold] QPCR analysis showed that CPD mRNA levels were not significantly increased in the benign prostate cell lines BPH-1 and RWPE1, as compared to a 3-4-fold increase in the pCa cell line LNCaP, following treatment with T or PRL (P[lt]0.01, n=3-6). T or PRL caused similar increases in other pCa cell lines (22Rv1, MDAPCa2b and PC-3), in which T and/or PRL receptors were expressed. DAF-2DA analysis revealed that NO levels were increased [lt]2-fold in benign cells, compared to [gt]3-fold in LNCaP cells after hormonal stimulation. T and PRL also showed a synergistic effect in upregulating NO levels in LNCaP pCa cells. Consistent with these results, TMA showed that CPD staining increased significantly from 8.9[plusmn]3.8% (mean[plusmn]SEM, n=18) of benign epithelial cell area to 30.9[plusmn]2.9% (n=79) of tumor cell area (P[lt]0.001). Endothelial cells in blood vessels associated with tumor were significantly more likely to show positive staining than vessels associated with benign tissue (P[lt]0.0001). Results from the TMA were corroborated by an IHC assessment of 26 large ([ge]50mm2) prostate specimens from men with cancer or benign prostatic hyperplasia.[br][bold]Conclusion:[/bold] T/PRL-stimulated CPD mRNA levels are higher in tumor than in benign prostate cell lines. Likewise, CPD protein levels are higher in cancer than benign prostate specimens. Increased CPD levels lead to an increase in NO production, which may contribute to pCa development. Our results implicate the potential usefulness of the CPD-Arg-NO pathway as a therapeutic target for pCa.[br][br](1) Thomas LN et al., Prostate 2012; 72:450.[br][br]Sources of Research Support: Study funded by Canadian Institutes of Health Research-RPP, Nova Scotia Health Research Foundation and Dalhousie Cancer Research Program (to CKLT).[br][br]Nothing to Disclose: LNT, CK-LT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2261 58 364 SAT-262 PO42-01 Saturday 303 2012


304 ENDO12L_SAT-263 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Prolactin and Androgen R1881 Upregulate Carboxypeptidase-D Gene Expression and Transcription for the Survival of Human Breast Cancer Cells Samir Koirala, Catherine KL Too Dalhousie University, Halifax, Canada; Dalhousie University, Halifax, Canada [bold]Introduction:[/bold] Carboxypeptidase-D (CPD) is a metalloproteinase that cleaves C-terminal arginine. Arginine is converted by nitric oxide synthase to nitric oxide (NO), which can stimulate cell growth. We have reported that 17[beta]-estradiol (E2) and prolactin (PRL) upregulated CPD mRNA/protein levels to increase NO production, which increased viability, and inhibited apoptosis of BCa cells (1, 2). The 2.0-kbp human CPD gene promoter contains a consensus interferon-[gamma]-activated sequence (GAS), several putative non-consensus androgen response elements (AREs), but no estrogen response element. The GAS and putative AREs may bind PRL-activated transcription factors Stat5a/Stat5b and the liganded androgen receptor (AR), respectively. The present study investigated regulation of CPD gene expression/transcription by E2, PRL, and synthetic androgen R1881, in BCa cells.[br][bold]Methods:[/bold] Quantitative RT-PCR (QPRC) and western analyses were used to measure CPD mRNA and protein levels, respectively, in BCa cells. CPD promoter constructs were transfected into BCa cells, and CPD promoter activity was measured using luciferase reporter assays.[br][bold]Results:[/bold] CPD mRNA and protein levels were upregulated by E2, PRL, and R1881, in a time- and dose-dependent manner, in MCF-7 and T47D BCa cell lines. The increase in CPD mRNA levels, after PRL or R1881 treatment, was supressed by actinomycin-D, suggesting transcriptional activation of the CPD gene. Consistent with this, PRL and R1881 stimulated the activity of a 2.0-kbp CPD promoter construct. When the two distal AREs (at -1994 and -1848 bp upstream from start site) were deleted from the 2.0-kbp construct, PRL or R1881 continued to stimulate promoter activity, suggesting that the GAS (at -1454) and third ARE (at -1443) responded to PRL and R1881, respectively. A 0.7-kbp CPD promoter construct lacking GAS and AREs was not activated by PRL nor R1881. In contrast, E2-stimulated CPD mRNA expression was not affected by actinomycin-D. Neither the 2.0-kbp nor 0.7-kbp promoter construct responded to E2. Thus, E2 may elicit CPD gene expression through non-genomic mechanisms, e.g., via protein kinase cascades.[br][bold]Conclusion:[/bold] PRL and R1881 stimulate CPD gene expression and transcription, whereas E2 stimulates CPD expression only. Since upregulation of CPD promotes NO production and BCa cell survival, our results implicate the potential usefulness of targeting the AR and PRL receptor signalling pathways in the treatment and/or management of BCa.[br][br](1) Abdelmagid SA and Too CKL, Endocrinology 2008; 149:4821. (2) Abdelmagid SA et al., J Cell Biochem 2011; 112:1084.[br][br]Sources of Research Support: SK is supported by a trainee award from the Beatrice Hunter Cancer Research Institute with funds provided by Cancer Care Nova Scotia as part of the Terry Fox Foundation Strategic Health Research Training Program in Cancer Research in CIHR. Study funded by CIHR-RPP, NSHRF, DCRP, and CBCF/Atlantic (to CKLT).[br][br]Nothing to Disclose: SK, CKLT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2253 58 365 SAT-263 PO42-01 Saturday 304 2012


305 ENDO12L_SAT-264 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) AKT Inhibitor, MK-2206, Reduces Leiomyoma Cell Viability and Induces Autophagy of Uterine Leiomyoma Cells [italic]In Vitro[/italic] and Reduces Tumor Volume [italic]In Vivo[/italic] Elizabeth C Sefton, Wenan Qiang, Vanida Serna, Takeshi Kurita, Julie Kim Northwestern University, Chicago, IL Uterine leiomyomas, benign tumors of the myometrium, are the number one indication for hysterectomies in the United States. Rates of surgery are high due to a lack of effective therapy. Uterine leiomyomas show increased phosphorylated AKT (p-AKT) compared to normal myometrium indicating that AKT signaling is increased in the tumor and may contribute to tumor viability. For this reason, we hypothesized that AKT inhibition would reduce leiomyoma cell viability. The AKT inhibitor MK-2206 was used to test our hypothesis. MK-2206 (MK) effectively reduced AKT and AKT substrate, PRAS40, phosphorylation over 24 hours. In addition, mTOR substrate p70S6K phosphorylation was reduced with MK. Although 10uM MK over 48 hours reduced leiomyoma cell viability, cell proliferation was unaffected. Despite that 10uM MK reduced viability, little apoptosis was induced as indicated by low cleaved caspase 3 induction and lack of annexin V staining coupled with DAPI positivity. Additionally, MK induced autophagy in leiomyoma cells as determined by LC3 II induction and punctate LC3 staining. Together these data indicate that apoptosis is not the major contributor to MK reduced cell viability and that autophagy may have a role in leiomyoma cell death. To determine if MK is effective in-vivo, primary human leiomyoma cells embedded in collagen were grown under the kidney capsule of immunocompromised mice and mice were treated weekly with 360 mg/kg MK for 3 weeks. MK reduced leiomyoma tumor volume in 2 of 3 patients tested compared to vehicle treated mice. Phosphorylated AKT was also reduced in lung tissue of MK treated mice compared to the control group indicating that MK was effective at the dose and treatment schedule. In summary we have demonstrated that AKT inhibition using MK reduces leiomyoma cell viability, induces autophagy, and reduces leiomyoma xenograft volume in-vivo. We are actively determining the function of MK induced autophagy in relation to leiomyoma cell survival and whether mTOR regulates autophagy in response to MK.[br][br]Sources of Research Support: NIH Grant P01HD057877-01A2.[br][br]Nothing to Disclose: ECS, WQ, VS, TK, JK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1699 58 366 SAT-264 PO42-01 Saturday 305 2012


306 ENDO12L_SAT-265 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Hyperactive Akt Signaling Causes Progestin Resistance in Endometrial Cancer through the Modulation of Progesterone Receptor B Transcriptional Function Irene I Lee, Julie Kim Northwestern University, Chicago, IL Progestin resistance presents the most significant challenge to improving fertility-sparing treatment in type I endometrial cancer. Currently, the response rates to conservative progestin therapy vary considerably and the molecular mechanisms behind progestin resistance are not well understood. Additionally, endometrial cancers can also be characterized in part by inactivating mutations in the PTEN gene; due to the loss of PTEN function, overactive Akt signaling persists in these cancers. Therefore, we hypothesized that in PTEN mutated endometrial cancers, hyperactive Akt signaling alters Progesterone Receptor B (PRB) transcriptional activity, leading to impaired progestin responses in endometrial cancer. Throughout our studies, we utilized the allosteric Akt inhibitor, MK-2206 (MK), to inhibit Akt activity in Ishikawa endometrial cancer cells. To determine the biological ramifications of combinatorial progestin (R5020) and Akt inhibitor (MK) treatments, we performed cell proliferation assays in PRB stably transfected Ishikawa cells and found that the combination of R5020 and MK inhibited cell proliferation more than either agent alone. In order to identify the specific PRB gene targets that are modulated by Akt signaling, we performed microarray gene expression analysis. Upon real-time PCR validation of target genes, we identified a subset of PRB target genes whose expression is dependent upon both Akt and PRB. However, not all PRB target genes were affected by Akt inhibition, indicating that the mechanism by which Akt modulates PRB target genes is context specific. We have identified [italic]PDK4[/italic] as a PRB target gene that is highly upregulated by combinatorial R5020 and Akt inhibition treatments. Additionally, we hypothesized that partnership with the PR cofactor, FOXO1, may suggest the mechanism by which inhibition of Akt affects expression of PRB target genes. We overexpressed a constitutively active form of FOXO1 (Tm-FOXO1) in PRB cells, and found that [italic]PDK4[/italic] expression is upregulated similarly to Akt inhibition treatment. In future studies, we plan to investigate the specific role that PDK4 plays in inhibiting cell proliferation in endometrial cancer as well as how FOXO1 may cooperate with PRB to govern the transcription of this gene as well as others. Taken together, these data suggest that Akt is sufficient to disrupt PRB transcriptional activity, and that inhibition of Akt may improve progestin responsiveness in endometrial cancer.[br][br]Sources of Research Support: NIH/NCI Training Grant T32CA09560 awarded to IIL; Malkin Scholars Program from the Robert H. Lurie Comprehensive Cancer Center of Northwestern University awarded to IIL; NIH R21 CA127674 awarded to JK.[br][br]Nothing to Disclose: IIL, JK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1872 58 367 SAT-265 PO42-01 Saturday 306 2012


307 ENDO12L_SAT-266 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Assessment of Steroid Hormones Upstream of P450c17 (CYP17) in Chemically Castrate Male Rhesus Monkeys Following Treatment with the CYP17 Inhibitors VT-464 and Abiraterone Acetate (AA) Joel R Eisner, David H Abbott, Ian M Bird, Stephen W Rafferty, William R Moore, Robert J Schotzinger Viamet Pharmaceuticals, Inc, Morrisville, NC; University of Wisconsin, Madison, WI; University of Wisconsin, Madison, WI CYP17 is a single protein with two distinct P-450-dependent enzyme activities, hydroxylase and lyase. Reduction of extra-gonadal androgen production through CYP17 inhibition is a validated paradigm for treatment of castration-resistant prostate cancer (CRPC) and may prove beneficial for the treatment of disorders of androgen excess, such as polycystic ovary syndrome. Treatment with the recently-approved CYP17 inhibitor, AA, increases overall survival in post-docetaxel CRPC patients, although due to potent CYP17 hydroxylase inhibition, is also associated with both cortisol (F) suppression and increased steroid hormone concentrations upstream of CYP17 [1,2]. The clinical candidate VT-464 is an oral, non-steroidal, lyase-selective CYP17 inhibitor. In vitro, VT-464 is approximately 10-fold more selective towards lyase than hydroxylase, while abiraterone is approximately 6-fold more selective towards hydroxylase than lyase. In a chemically-castrate monkey model, both single-dose VT-464 and AA significantly reduced plasma androgen concentrations, but AA treatment also resulted in significant F suppression and progesterone (P4) elevation [3]. The objective of the current study was to compare the upstream steroid hormone profiles in adult, chemically-castrate male rhesus monkeys following treatment with VT-464 and AA, respectively. LC/MS/MS methods were used to measure plasma P4, pregnenolone (P5), corticosterone (CS) and deoxycorticosterone (DOC) concentrations 4 hours after a single subcutaneous dose (6.25 mg/kg) of VT-464, AA, or vehicle. Treatment with AA resulted in an increase in all four upstream steroid hormones compared to VT-464 and vehicle treatments, respectively: P4 (29.7 [plusmn] 7.2 SEM vs. 0.7 [plusmn] 0.2 and 1.4 [plusmn] 0.8), P5 (16.1 [plusmn] 3.5 vs. 0.6 [plusmn] 0.1 and 2.3 [plusmn] 0.4), CS (283.8 [plusmn] 22.3 vs. 35.8 [plusmn] 6.4 and 100.3 [plusmn] 17.5), and DOC (4.9 [plusmn] 1.2 vs. 0.5 [plusmn] 0.1 and 0.7 [plusmn] 0.2). The lack of increased upstream steroid hormone concentrations following VT-464 treatment, in monkeys that previously accumulated such steroid hormones following AA treatment, confirms VT-464 lyase-selectivity. VT-464, which is in early clinical trials for CRPC, may provide suppression of extra-gonadal androgens without mineralocorticoid-induced side effects (CS and DOC excess) or elevation of P4 and P5, which have been implicated in activating the mutated androgen receptors found in some CRPC patients.[br][br][1] de Bono et al, N Engl J Med 2011; 364:1995-2005. [2] Attard et al, J Clin Endocrinol Metab 2012;97(2). [3] Eisner et al, Genitourinary Cancers Symposium, San Francisco, CA: 2012. Abstract No. 198.[br][br]Disclosures: JRE: Employee, Viamet Pharmaceuticals, Inc. DHA: Ad Hoc Consultant, Viamet Pharmaceuticals, Inc.; Recipient Award, Viamet Pharmaceuticals, Inc. IMB: Consultant, Viamet Pharmaceuticals, Inc. SWR: Employee, Viamet Pharmaceuticals, Inc. WRM: Employee, Viamet Pharmaceuticals, Inc. RJS: Employee, Viamet Pharmaceuticals, Inc. 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1852 58 368 SAT-266 PO42-01 Saturday 307 2012


308 ENDO12L_SAT-267 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Mechanisms of Intrinsic Transactivation of PPAR[gamma]1 in Breast Cancer Progression Natalie Wallis, Linah Al-Alem, Yekaterina Zaytseva, Diana Syam, Michael Kilgore University of Kentucky College of Medicine, Lexington, KY; University of Kentucky College of Medicine, Lexington, KY; University of Kentucky College of Medicine, Lexington, KY Expression of the peroxisome proliferator-activated receptor gamma 1 (PPAR[gamma]1) and its intrinsic activation play critical roles in breast cancer progression yet little is known of the mechanisms controlling these events. PPAR[gamma]1 is highly overexpressed in human breast cancer including triple negative breast cancer and in a genetic model of constitutive transactivation PPAR[gamma] drives promotion, increases the rate of metastasis and greatly reduces survival. By contrast, synthetic ligands are reported to be antiproliferative in breast cancer cells though it is unclear whether these actions are mediated through PPAR[gamma]1. Furthermore, intrinsic activation of PPAR[gamma]1 accompanies the increase in expression seen in malignant transformation. Both are necessary for the survival as the knockdown of PPAR[gamma]1 is lethal in breast cancer cells. While these studies clearly demonstrate that endogenous transactivation is critical to breast cancer progression the mechanism(s) controlling its activity remain poorly understood. Neither the identity of the endogenous ligand(s) nor the contributions that post-translational modifications (PTMs) play in tumor progression are known. To define the nature and location of the PTM made to PPAR[gamma]1 during tumor progression we have constructed both HA- and FLAG-tagged expression vectors of wild type PPAR[gamma]1. These are being used in proteomic approaches to compare the PTMs seen between normal mammary epithelial cells (HMEC), non-transformed cell lines (MCF10A) and metastatic breast cancer (MDA-MB-231) cells. Additionally, site directed mutations to putative phosphorylation (S84A) and sumoylation sites (K79R and K367R) have also been constructed. The ability of these to control transcription demonstrates an interdependence of the PTM and the response to the exogenous ligand pioglitazone. It is equally possible the intrinsic activation of PPAR[gamma]1 is mediated by the synthesis of an endogenous ligand; therefore, we have also constructed a mutation in the ligand-binding domain (Q286P) that prevents ligand activation of transcription but does not alter basal activity. The fact that Q286P does not act as dominant negative suggest that an endogenous ligand may not be present although additional studies will be required to verify this observation. Together, these studies could explain the [ldquo]Janus Faced[rdquo] actions of PPAR[gamma]1 in cancer progression and reveal novel therapeutic strategies for treatment.[br][br]Sources of Research Support: NCRR-P20-RR1559 and R01 CA95609.[br][br]Nothing to Disclose: NW, LA-A, YZ, DS, MK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 947 58 369 SAT-267 PO42-01 Saturday 308 2012


309 ENDO12L_SAT-268 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) The Group II Metabotropic Glutamate Receptor 3 (GRM3) Expression in Human Prostate Cancer: A Possible Regulator of Microenvironment of Cancer Tissue Yasuhiro Nakamura, Sauro Ferizola, Shunichi Takeda, Takashi Maekawa, Kazue Ise, Shigeto Ishidoya, Yoichi Arai, Hironobu Sasano Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku University Graduate School of Medicine, Sendai, Japan [bold]Background[/bold]: G protein-coupled receptors (GPCRs) are well-known as one of the important regulators of cell signaling pathways. Among GPCRs, the group II metabotropic glutamate receptor (GRM3) negatively modulates adenylate cyclase signaling cascade and regulates neurotransmission in human brain. GRM3 expression is also reported in various types of human malignancies including glioma, ganglioglioma, laryngeal cell carcinoma and adrenocortical tumor and considered a key regulator of cell proliferation in these cancers. Estrogens have been also reported to activate GRM3 signaling, resulting in suppression of cAMP response element-binding protein in cultured rat hippocampal neurons. Both androgens and estrogens play important roles in the pathogenesis of prostate cancer and we previously reported that the [italic]in situ[/italic] production of these hormones has been proposed to play a critical role in the pathogenesis and/or development of human prostate cancer. However it remains unclear whether GRM3 is expressed or not in human prostate cancer tissue.[br][bold]Experimental design[/bold]: We evaluated GRM3 expression in human prostate cancer obtained from surgery (n=112) using immunohistochemical analysis, and correlated the findings with clinicopathological features of the patients and status for steroidogenic enzymes in these specimens.[br][bold]Results[/bold]: GRM3 immunoreactivity was detected in 36 cases (32%) and was significantly correlated with that of Gleason score ([italic]P[/italic]=0.018), immunoreactivity of 17-beta-hydroxysteroid dehydrogenase type 1 (17bHSD1) ([italic]P[/italic]=0.015) and aromatase ([italic]P[/italic]=0.005).[br][bold]Conclusion[/bold]: These data all suggest that GRM3 expression level is correlated with histological grade of prostatic carcinoma cells and possibly [italic]in situ[/italic] estrogen-production in human prostate cancer tissues and GRM3 could be a novel target for hormonal manipulation of human prostate cancer.[br][br]Nothing to Disclose: YN, SF, ST, TM, KI, SI, YA, HS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 107 58 370 SAT-268 PO42-01 Saturday 309 2012


310 ENDO12L_SAT-269 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Amino-Terminal Segment of Aromatase Possesses Distinct Functional Domains for Its Targeting to Endoplasmic Reticulum Jasmeet Kaur, Ray M Rudolph, Himangshu S Bose Mercer University School of Medicine, Savannah, GA; Memorial University Medical Center, Savannah, GA Estrogen plays fundamental role in breast cancer as approximately 70% breast tumors express estrogen receptor. In estrogen biosynthesis, aromatase catalyzes the last step, thereby producing estrogens from androgens. Over-expression of this enzyme leads to overproduction of estrogen in breast cancer tissue. Thus, inhibiting estrogen through impeding aromatase is more effective endocrine treatment than blocking its action. Aromatase is an endoplasmic-reticulum (ER)-resident protein, so we expect a different mechanism for its translocation and in turn, accumulation in normal and affected tissue. So, we propose that aromatase has specific conformational domains that can be potential targets to combat its overexpression. Our results show that aromatase strongly follows the co-translocation mechanism to process into ER and is a glycoprotein. It possesses type I-like transmembrane topology with N-terminal on the non-cytoplasmic side and C-terminal in the reducing cytosol. It is strongly integrated into the membrane through its N-terminal. N-terminal works as an uncleavable signal sequence as well as stop-transfer/membrane-integration signal. In the absence of the specific N-terminal sequence, the protein is not glycosylated. Further, this sequence is a little downstream unlike other cytochrome P450s[apos] amino terminal. Thus, the conformation of this protein is extremely specific and important for its targeting and localization into the ER and can be exploited for better targeted drug development against breast cancer.[br][br]Nothing to Disclose: JK, RMR, HSB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 426 58 371 SAT-269 PO42-01 Saturday 310 2012


311 ENDO12L_SAT-270 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Role of Calcium in Hormone-Resistant Breast Cancer Kedra Cyrus, Mudit Kaushal, Fatima Gibrel, Anna Chichura, William Yeguech, Mary Beth Martin Georgetown University, Washington, DC; University of the District of Columbia, Washington, DC; Montgomery College, Rockville, MD Central to the growth and progression of a subset of breast cancer is the estrogen receptor alpha (ER[alpha]). As such, use of antiestrogens has become the standard of care in the treatment of ER[alpha] -positive breast cancer. Unfortunately, one-third of patients receiving antihormone treatment eventually develop resistance to therapy. There are several factors contributing to resistance, including increased hormone independent activation of the ER[alpha] by signal transduction pathways, due to overexpression of receptors such as the epidermal growth factor (EGF) receptor. Recently, our laboratory has shown that calcium mediates the cross talk between EGF and ER[alpha] by directly binding to and activating the ligand binding domain of the ER[alpha]1. These data suggest that calcium is a legitimate ligand for ER[alpha]. The calcium mediated EGF- ER[alpha] cross talk may also hint at a role for calcium in the development and progression of hormone resistance and may provide a new target for breast cancer therapy. Using a hormone resistant breast cancer cell line LCC-9, we investigated the effects of FDA approved calcium channel blockers on the growth of these resistant cells, as well as the effects on gene transcription. The intracellular calcium chelator BAPTA-AM was also used to query the effects of blocking intracellular calcium in these cells. Our preliminary results suggest that chelation of intracellular calcium and the combined use of calcium channel blockers with antiestrogens results in the restoration of hormone sensitivity in these cells, pointing to a role for calcium in hormone resistance.[br][br]1Divekar , SD et al. The Role of Calcium in the Activation of Estrogen Receptor [ndash]Alpha Cancer Res. 2011 Mar 1;71(5):1658-68.[br][br]Nothing to Disclose: KC, MK, FG, AC, WY, MBM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2186 58 372 SAT-270 PO42-01 Saturday 311 2012


312 ENDO12L_SAT-271 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Three-Dimensional (3D) Micro-Environments May Provide Cell Templates for Drugs That Are Different from Those of Two Dimensions (2D) Kenny Chitcholtan, Peter H Sykes, John J Evans University of Otago, Christchurch, New Zealand; University of Otago, Christchurch, New Zealand One aspect of local cell environment is the physical characteristics, such as provided by two dimension (2D) monolayer cultures in vitro, which will influence the function of cells. Thus three-dimension (3D) models may provide more realistic recapitulation of in vivo states. We compared endometrial cancer cell lines in a 3D model, using a reconstituted basement membrane (3D rBM) (GelTrex[trade]), and 2D monolayer in vitro cell culture. 3D rBM strikingly reduced cell proliferation compared with 2D cell culture. Thus drugs tested on rapidly proliferating cells in 2D may be assigned properties that would be unreliably translated. The morphologies of the resultant 3D entities were determined. Ishikawa and RL95-2 cell lines formed cell clusters of spherical morphology in 3D culture. KLE and EN1078D cell lines formed disorganised tumour sheets and multicellular spheroids, respectively. After 8 days in 3D rBM Ishikawa cell clusters possessed central lumen, which in RL95-2 cells were less well-defined. Such interactions between morphology and growth may be dependent on integrins. This investigation revealed that [beta]4-integrin localised to the basal region of polarised Ishikawa cells but it was observed within clusters of poorly differentiated cell lines. In addition, RL95-2, KLE and EN1078D tended to have higher expression of [beta]4-integrin subunit than [beta]1-integrin. [beta]1-integrin was localised to regions where cancer cells were in contact with the rBM. Ishikawa cells increased [beta]4 and [beta]1-integrin expression in 3D compared to 2D culture. Secretion of an end-product, VEGF, was markedly reduced in Ishikawa and KLE cell lines grown in 3D rBM, whereas there was no difference in RL95-2 and EN1078D cancer cells. In investigation of underlying intracellular signalling cascades we noted that the addition of exogenous EGF to endometrial cancer cells in 3D rBM activated p-EGFR and p-Akt in both cell culture conditions. Investigation of the different structures noted that glucose metabolism was selectively reduced in 3D of Ishikawa and KLE cells compared with 2D cell culture. In contrast EN1078D cells increased glucose metabolism in 3D culture. There was no difference in Glut-1 expression in cells cultured in 3D rBM or 2D cell culture. Taken together, our data demonstrated that 3D rBM induces differences in endometrial cancer cell behaviour compared to 2D cell monolayer counterparts, and it may provide more suitable in vitro models to understand the tumorigenesis of endometrial cancer.[br][br]Nothing to Disclose: KC, PHS, JJE 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1218 58 373 SAT-271 PO42-01 Saturday 312 2012


313 ENDO12L_SAT-272 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) 17[beta]-Estradiol Prevents Taxol-Induced Apoptosis in Triple-Negative Breast Cancer Cells Reyhaan A Chaudhri, Agreen Hadadi, Zvi Schwartz, Barbara D Boyan Georgia Institute of Technology, Atlanta, GA; Georgia Institute of Technology, Atlanta, GA Hormone effects in breast cancer can vary among patients. However, in most cases where estrogen is concerned, tumor growth is promoted and this may be through non-classical effects of estrogen from the estrogen receptors (ERs) on the plasma membrane. The effects of estrogen can inhibit the actions of taxol, a commonly used chemotherapeutic drug used to induce apoptosis in many types of cancer, including breast cancer. In both ER-positive and ER-negative breast cancer cells, we previously found that 17[beta]-estradiol (E2) can enhance cellular proliferation, while promoting anti-apoptotic effects through the membrane receptor, ER[alpha]36. The mechanism by which this occurs is not fully understood. The purpose of this study was to elucidate the mechanism by which breast cancer tumorigenicity is enhanced by E2. We hypothesized that membrane-associated E2 signaling via ER[alpha]36 leads to activation of a pathway that may include specific mediators, such as G-proteins, phospholipase D (PLD), phospholipase C (PLC), protein kinase C (PKC), phosphatidylinositol-3 kinase (PI3K), cytochrome C, and others that allow E2 signaling to inhibit taxol-induced apoptosis in breast cancer cells. In order to determine which proteins play a role in anti-apoptotic signaling, triple negative HCC38 breast cancer cells were divided into 4 different treatment groups (Control, E2-treated, taxol-treated, and E2/taxol-treated). Each group was treated with and without inhibitors of the aforementioned signaling molecules. After 4 hours, caspase-3 activity was measured and after 24 hours, cell viability was determined by the MTT assay. Previously we have shown that taxol increases caspase-3 activity and lowers cell viability, while E2 inhibits this. In this study, E2 rapidly activated PKC and PLD activity, and inhibitors to PKC, PLC and PI3K blocked E2[apos]s effect on taxol-induced apoptosis. Antibodies to ER[alpha]36 also blocked the effect of E2 on preventing taxol-induced apoptosis. Based on these data, we can conclude that the pathway by which E2 prevents taxol-induced apoptosis is activated by ER[alpha]36 on the plasma membrane, and includes activation of PLC, PLD, PI3K, and PKC.[br][br]Sources of Research Support: The Price Gilbert Jr. Foundation and by PHS Grant UL1 RR025008 and TL1 RR025010 from the Clinical and Translational Science Award program, National Institutes of Health, National Center for Research Resources.[br][br]Nothing to Disclose: RAC, AH, ZS, BDB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2230 58 374 SAT-272 PO42-01 Saturday 313 2012


314 ENDO12L_SAT-273 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Steroidogenic Factor 1 Expression Induces Steroidogenic Enzyme Expression and Steroid Synthesis To Fuel Growth of Aggressive Prostate Cancer Cells Samantha R Lewis, Joan S Jorgensen University of Wisconsin, Madison, WI The prostate depends on androgens synthesized by the testis both in disease and development. Although androgen deprivation therapy by medical or surgical castration is a cornerstone of treatment for metastatic prostate cancer, remission is only temporary and tumors inevitably progress to castration refractory prostate cancer. There are several theories to explain this aggressive resurgence, one of which suggests that prostate adenocarcinoma cells acquire machinery for de novo steroidogenesis. During development, and throughout life, Steroidogenic Factor 1 (NR5A1, ADBP4, SF1) is a key regulator of steroidogenesis in normal endocrine tissues. Normal and benign prostate tissues lacks SF1 expression and are non-steroidogenic. Therefore, we hypothesize that SF1 is expressed in castration refractive prostate cancer to stimulate steroidogenesis and fuel malignant growth. We examined SF1 expression in a panel of prostate epithelial cell lines and found SF1 present in each aggressive cancer cell line examined but absent in benign prostate cell lines. These results were supported by detection of SF1 in human prostate cancer tissue specimens from patients with castration refractory prostate cancer, whereas it is conspicuously absent from all specimens of localized prostate cancer and benign prostate thus far examined. To functionally link SF1 to steroidogenic production in prostate epithelial cells, we tested whether ectopic expression of SF1 was sufficient to induce steroid production in a benign prostate epithelial cell line (BPH-1). BPH-1 cells expressing ectopic SF1 exhibit increased mRNA levels of steroidogenic enzymes and secrete estradiol, which was inhibited by treatment with the aromatase inhibitor, anastrozole. These studies were followed by examining effects of shSF1 in an SF1-expressing cell line of malignant transformed prostate epithelial cells that synthesize estradiol in levels similar to SF1-transfected BPH-1 cells. shSF1-transfecting cells resulted in decreased estradiol production and a slower proliferation rate. Based on these data, we conclude that SF1 is induced in aggressive prostate cancers and is sufficient to stimulate steroidogenesis to promote aggressive growth. Our findings present a new potential mechanism and therapeutic target for deadly prostate cancer.[br][br]Sources of Research Support: The University of Wisconsin-Madison and NIH 5T32-HD041921-07.[br][br]Nothing to Disclose: SRL, JSJ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1712 58 375 SAT-273 PO42-01 Saturday 314 2012


315 ENDO12L_SAT-274 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Opposing Effects of Estrogen Receptor [beta]1 and [beta]2 in the Prostate Cancer Cell Line PC3 Prasenjit Deay, Jan-Ake Gustafsson, Anders Strom University of Houston, Houston, TX; Karolinska Institutet, Stockholm, Sweden Estrogen receptor [beta]1[apos]s (ER[beta]1) expression is lost during the progression of prostate cancer and cannot be detected beyond Gleason grade 3. In agreement with clinical studies indicating a tumor suppressive effect of ER[beta] we find that expression of the receptor has anti-proliferative effect in the prostate cancer cell line PC3. A splice variant of ER[beta]1; ER[beta]2 was discovered two years after the parent receptor, but surprisingly very little is known about this splice variant. We describe that this variant has opposing effect to ER[beta]1 on proliferation in PC3 cells. Furthermore, we found that the c-myc protein is down regulated by ER[beta]1 but is up regulated on both mRNA and protein level by ER[beta]2. In addition, we found that Twist1, a factor involved in prostate cancer metastasis is strongly up regulated by ER[beta]2 but not changed by ER[beta]1. It is interesting to note that increased expression of both c-myc and Twist1 correlates to worse prognosis of prostate cancer, indicating a tumor suppressive effect of ER[beta]1 and a tumor promoting effect of ER[beta]2. This is furthermore in agreement with a recent finding that expression of ER[beta]2 in metastatic prostate cancer correlates with reduced overall survival. We also found that a dominant negative ras is blocking ER[beta]2[apos]s enhancement of the Twist1 promoter activity in PC3 cells indicating that ER[beta]2[apos]s tumor promoting effect is dependent on ras signaling.[br][br]Sources of Research Support: The Cancer Prevention [amp] Research Institute of Texas (CPRIT) grants HIRP100680 and RP110444, the Texas Emerging Technology Fund under Agreement no. 300-9-1958, the Robert A. Welch Foundation and the Swedish Cancer Fund. All awarded to JAG.[br][br]Nothing to Disclose: PD, J-AG, AS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2038 58 376 SAT-274 PO42-01 Saturday 315 2012


316 ENDO12L_SAT-275 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) ER[beta]1 Regulates the XBP-1 Pathway of the Unfolded Protein Response in Breast Cancer Cells Gayani Rajapaksa, Fotis Nikolos, Igor Bado, Christoforos Thomas, Jan-Ake Gustafsson University of Houston, Houston, TX Estrogen-regulated transcription factors are important in the physiology of the mammary gland as well as in the progression and anti-estrogen resistance of breast cancer. Endocrine therapy resistance is a significant challenge in the treatment and disease management of breast cancer. Unfolded protein response (UPR) has been identified as one of the key mechanisms determining endocrine responsiveness and cellular survival in breast cancer. UPR assists the cells to cope the stress caused by unfolded proteins in the endoplasmic reticulum. Although estrogen receptor [alpha] (ER[alpha]) has been shown to modulate the UPR-dependent endocrine resistance, the role of wild-type ER[beta] (ER[beta]1) in UPR has not been elucidated.[br]Control and ER[beta]1-expressing MDA-MB-231 and MCF7 breast cancer cells were treated with pharmacological stress inducer-thapsigargin and subjected to FACS analysis, protein and RNA profiling.[br]Here we show that ER[beta]1 decreased the resistance of breast cancer cells to ER stress by altering the inositol-requiring enzyme-1 (IRE-1) pathway of the UPR. Expression of ER[beta]1 repressed the mRNA splicing of X-box binding protein-1 (XBP-1) that correlates with endocrine resistance and survival in breast cancer cells. Furthermore ER[beta]1 induced apoptosis in breast cancer cells by repressing the IRE-1-mediated expression of Bcl-2 and promoting the cleavage of caspace 3. These results suggest the potential role of ER[beta]1 in regulating resistance of breast cancer to variety of stressors including anti-estrogens through regulation of the unfolded protein response.[br][br]Nothing to Disclose: GR, FN, IB, CT, J-AG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1965 58 377 SAT-275 PO42-01 Saturday 316 2012


317 ENDO12L_SAT-276 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) 1[alpha],25-Dihydroxyvitamin D[sub]3[/sub] Inhibits Nandrolone-Stimulated Proliferation in Human Prostate LNCaP Cells Helenius J Kloosterboer, Marjanne Prins, Reiner Class KC2, Oss, Netherlands; OrgaNext, Arnhem, Netherlands; Pharmacelsus, Saarbruecken, Germany [bold]Introduction. [/bold]Nandrolone decanoate has a high selectivity index with respect to anabolic effects on muscles over androgenic effect on the prostate in vivo. Previously, we have shown that nandrolone combined with 1[alpha],25-dihydroxyvitamin D[sub]3[/sub] (DHVD[sub]3[/sub]) has a synergistic effect on human muscle cell proliferation. The aim of our study was to investigate whether DHVD[sub]3[/sub] influences the effect of nandrolone on the proliferation of human prostate LNCaP cells. We also determined in LNCap cells the effect on the S-phase of the cell cycle.[br][bold]Materials and Methods.[/bold] LNCaP cells were grown in RPMI 1640 medium supplemented with 10% Fetal Bovine Serum (FBS). Before the cells were seeded in 96-well plates medium was changed daily for 5 days with medium plus 10% charcoal-stripped FBS (CCS-FBS). Cells were trypsinized and seeded in 96-well plates at a density of 3000 cells per well. After 48 hrs, nandrolone and reference testosterone (T) were added over the concentration range of 10[sup]-14[/sup] to 10[sup]-6[/sup] M either alone or in the presence of 10 nM DHVD[sub]3[/sub]. After 48 hrs, medium containing compounds was replaced and 72 hrs later, cell growth was quantified by the resazurin assay. LNCaP cells to be used for cell cycle experiments were grown in 25 cm[sup]2[/sup] flasks under the same conditions except that the cells were seeded in FBS-containing medium, followed by 24 hrs in CCS-FBS. Subsequently, nandrolone (10-1000nM) and T (1-100nM) were added either alone or in combination with 10 nM DHVD[sub]3[/sub]. Medium containing compounds was refreshed after 24 hrs and 24 hrs later the % cells in G1/0-phase, S-phase and G2/M-phase were detemined by EdU incorporation and propidium iodine counter staining. Analysis was done using a dual laser flow cytometer (Beckman Coulter).[br][bold]Results.[/bold] Nandrolone has a 3 times higher binding to androgen receptor than T, but the growth curve for nandrolone showed a lower maximal response than that for T. DHVD[sub]3[/sub] alone has little or no effect on growth of LNCaP cells. Combining nandrolone with 10 nM DHVD[sub]3[/sub] reduced the growth with about 25%. Cell cycle experiments showed that nandrolone increased the % of cells in the S-phase with 3-4% compared to control, 3 times less compared to the reference T. DHVD[sub]3[/sub] had a neglectable effect.[br][bold]Conclusions.[/bold] The combination of DHVD[sub]3[/sub] and nandrolone resulted in 25% lesser growth of LNCaP cells than with nandrolone alone. Combining these results with the effects on muscle cells it can be concluded that 1[alpha],25-dihydroxyvitamin D[sub]3[/sub] improves the selectivity index for nandrolone.[br][br]Nothing to Disclose: HJK, MP, RC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1701 58 378 SAT-276 PO42-01 Saturday 317 2012


318 ENDO12L_SAT-277 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Circulating Stem Progenitor Cells and Mammographic Density in Postmenopausal Females Receiving Metformin and N-Acetylcysteine Lev M Berstein, Dmitry A Vasilyev, Natalya V Bychkova NN Petrov Research Institute of Oncology, St. Petersburg, Russian Federation Objectives: Mammographic density (MD) increase is an important breast cancer (BC) risk factor. Association between MD value and progenitor stem cells circulating in blood (cSC) has not been studied. cSC fraction CD44 belonging to mammary stem cells correlates with BC stage (Baek et al., 2011) while number of circulating CD34 cells which is increased in BC patients (Richter-Ehrenstein et al., 2007), according to some data decreases in impaired glucose tolerance and metabolic syndrome (Fadini et al., 2007). Materials: Following over-night fast, blood levels of CD34+, CD44+, CD90+ (a tumor suppressor for some tumors, Abeysinghe et al., 2003) and CD44+CD90+ cells were evaluated using Cytomics FC 500 (Beckman-Coulter) flow cytometer in 34 cancer-free postmenopausal females aged 56,1[plusmn]0,7. Received data were confronted with recently assessed MD value (mean 22,5[plusmn]3,5%), signs of glucose intolerance and metabolic syndrome and with response of MD in 17 of the studied women to long-term, not less than 6 mo, treatment with antidiabetic biguanide metformin, 1500 mg/day or N-acetyl cysteine, 600 mg/day. Results: Average fasting levels of cSC were the following (cond. units): CD34 0,072[plusmn]0,007, CD44 95,94[plusmn]0,59, CD90 0,326[plusmn]0,044 and CD44/CD90 double positive 0,311[plusmn]0,042. In the whole studied group, a tendency to negative Spearman rank correlation was revealed between MD value and CD90 level (R -0,236, p 0,23), while in females with glucose intolerance (R -0,34, p 0,11) and metabolic syndrome (R -0,46, p 0,07) this association was pronounced more distinctly. In regard of CD34 cells, the negative correlation of their level with MD was characteristic only for women with metabolic syndrome (R -0,40, p 0,09) but not for females with impaired glucose tolerance. Content of CD44 had been associated with MD value positively and similarly in cases with glucose intolerance (R 0,54) and metabolic syndrome (R 0,55, p 0,11) while this was not peculiar to females without such metabolic disturbances. No changes of cSC levels were discovered after metformin and N-acetyl cysteine course in spite of the decrease of MD (- 3,2% and -10,0% in mean absolute and relative values respectively) in 8 of 17 treated women. Conclusions: An association of cSC with MD value is determined by properties of these cells as well as by metabolic phenotype of postmenopausal females while moderate decrease of MD under the influence of metformin and N-acetyl cysteine is not mediated through cSC involvement.[br][br]Baek et al., Breast Cancer Research Treatment 2011;130:1029. Richter-Ehrenstein et al., Breast Cancer Research Treatment 2007;106:343. Fadini et al., Diabetologia 2007; 50:2156. Abeysinghe et al.,Cancer Genet Cytogenet 2003;143:125.[br][br]Sources of Research Support: RFBR grant 12-04-00084.[br][br]Nothing to Disclose: LMB, DAV, NVB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 149 58 379 SAT-277 PO42-01 Saturday 318 2012


319 ENDO12L_SAT-278 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Metformin Does Not Decrease Immunohistochemical Aromatase Expression in Breast Tumors of Diabetic Patients Lev M Berstein, Elena A Turkevich, Marina P Boyarkina NN Petrov Research Institute of Oncology, St. Petersburg, Russian Federation Objectives: Estrogens are important for, at least, some breast tumors. Although others have demonstrated that antidiabetic biguanide metformin (MF) inhibits aromatase expression in certain cell types in vitro (Takemura et al., 2007; Rice et al., 2009; Brown et al., 2010), to the best of our knowledge, data about the intratumoral aromatase expression in breast cancer (BC) of diabetic patients and on the possible consequences of antidiabetic therapy preceding cancer diagnosis are lacking. Materials: Tumor paraffin blocks were obtained from 57 pts aged 48-77 yrs (mostly ductal invasive BC with moderate to high grade of malignancy, [gt]80% at stages T1-2N0-3M0; 13 pts without diabetes, and 44 with type 2 diabetes with following treatment modalities for 1 yr at least: diet only (n=14), sulphonylurea (SU, n=14), metformin (MF, n=9), and 7 MF+SU (n=7)). Four-micron sections deparaffinized in xylene were processed with the monoclonal aromatase antibody 677. Four to five different fields from each section were analyzed and evaluated semi-quantitatively for the proportion of positively stained cells and the intensity of staining using conventional scores to distinguish four categories: negative - 0, weak - 1, moderate - 2, and strong - 3. Negative controls were processed with 0.01 M PBS instead of the specific antibody. Sections from paraffin-embedded human placenta blocks were used as a positive control. Results: Mean conventional scores that combine data on the percent of aromatase immunopositive cells and on the intensity of staining were as follows: 1.31 (no diabetes), 1.47 (all diabetic patients), 2.22 (MF), 1.50 (SU), 1.29 (MF+SU), 1.81 (MF and MF+SU), 1.07 (diet). Allred scores for progesterone receptor (PR) were highest in the tumors of diabetics treated with MF or MF+SU and lowest in the tumors of SU-treated pts. Conclusions: Thus, in contrast to previous findings that suggested the suppressive effect of MF on aromatase, no such trend was discovered for aromatase expression in tumors of diabetic patients treated with MF. Although this observation is still needs to be confirmed using a more representative group of patients and possibly another aromatase antibody, together with the data evidencing higher PR levels (see also: Berstein et al., Med. Oncol. 2011) this may suggest better responses to hormonal therapy in MF-treated patients and supports hopes for MF usefulness in breast cancer treatment.[br][br]Takemura et al., J Clin Endocrinol Metab. 007;92:3213. Rice et al., Endocrinology 2009; 150:4794. Brown et al., Breast Cancer Res Treat. 2010; 123:591. Berstein et al., Med Oncol. 2011;28:1260.[br][br]Sources of Research Support: Acknowledgements. To Drs. Dean Evans and Dick Santen for help in obtaining of aromatase antibody 677.[br][br]Nothing to Disclose: LMB, EAT, MPB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 147 58 380 SAT-278 PO42-01 Saturday 319 2012


320 ENDO12L_SAT-279 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Presence of Estrogen Receptor-[alpha]36 in ER[alpha]-Positive and ER[alpha]-Negative Breast Cancer Reyhaan A Chaudhri, Natalia Cuenca, Ruth O[apos]Regan, Barbara D Boyan, Zvi Schwartz Georgia Institute of Technology, Atlanta, GA; Georgia Institute of Technology, Atlanta, GA; Emory University, Atlanta, GA Breast cancer treatment is determined primarily by the expression of hormone receptors and HER2. Traditionally, ER-positive cancers are felt to depend on estrogen for growth, while ER-negative cancers do not. However, we previously found that ER-negative breast cancer cells not only respond to estradiol, but actually harbor a recently-identified alternatively spliced variant of ER[alpha], ER[alpha]36. Through this receptor that resides in the plasma membrane, estrogen can illicit proliferative and anti-apoptotic effects. The purpose of this study was to evaluate the presence of ER[alpha]36 in vivo using immunohistochemistry on a breast tissue microarray containing 36 tumor sections from different patients. Of these 36 tumor sections, we found that 16 were positive for ER[alpha]36 ([sim]44%) of which 5 were classified as luminal A (ER+/PR+/HER2-), 4 were classified as triple negative (ER-/PR-/HER2-), 3 were ER- (ER-/PR+/HER2+), 2 were luminal B (ER+/PR+/HER2+), and 2 were HER2+ (ER-/PR-/HER2+). We also noticed that in the majority of samples, ER[alpha]36 staining appeared non-nuclear, and in many cases where vasculature was present in the tissue section, we noticed that red blood cells were strongly positive. This supports our previous in vitro data that ER[alpha]36 functions primarily as a non-nuclear estrogen receptor. Of further note is that 9 of the 16 ER[alpha]36 positive samples were from patients of a pre-menopausal age group ([lt]50 years of age), while the rest of the positive samples (7) were from patients in a post-menopausal age group ([gt]50 years of age). As far as nodal status of the tumor is concerned, 7 samples were from patients with regional lymph node metastasis, while 9 patients with positive staining did not show regional lymph node metastasis or the lymph node status was not available. Fisher[apos]s exact test was used to obtain p-values for the following associations: ER-status versus ER[alpha]36 status (p=0.7830); menopausal status versus ER[alpha]36 status (p=0.3437); nodal status versus ER[alpha]36 status (p=0.7106). Our results show no significant association among these variables; however, we can conclude that ER[alpha]36 status is independent of these variables, and therefore may possess unique functions than typical ERs. Further functional analysis is necessary to evaluate this hypothesis.[br][br]Sources of Research Support: The Price Gilbert Jr. Foundation and by PHS Grant UL1 RR025008 and TL1 RR025010 from the Clinical and Translational Science Award program, National Institutes of Health, National Center for Research Resources.[br][br]Nothing to Disclose: RAC, NC, RO, BDB, ZS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1985 58 381 SAT-279 PO42-01 Saturday 320 2012


321 ENDO12L_SAT-280 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Resveratrol Negatively Influences NAMPT in a Hepatocarcinoma Cell Line without Modifying Intracellular NAD Levels [mdash] Does Nicotinamide Play a More Important Role? Susanne Schuster, Stefanie Petzold-Quinque, Antje Garten, Melanie Penke, Anja Barnikol-Oettler, Sandy Laue, Rolf Gebhardt, Wieland Kiess University Hospital Leipzig, Hospital for Children and Adolescents, Leipzig, Germany; University Leipzig, Medical Faculty, Leipzig, Germany Background: Nicotinamide phosphoribosyltransferase (NAMPT) is a key enzyme of NAD biosynthesis and regulates the activity of the NAD- dependent deacetylase sirtuin 1 (SIRT1). SIRT1 is implicated in modulating cellular energy metabolism, stress response and multiple aging-related diseases including cancer. Resveratrol (Resv), a natural dietary polyphenol found in grape skin and red wine is a potential SIRT1 activator and has been proven to be an effective chemopreventive agent. We hypothesized that NAMPT acts as key player in the Resv-mediated apoptosis of tumoral cells through a reduced metabolisation of nicotinamide.[br]Results: Resv reduced cell viability by 44.8[plusmn] 5.3% (100[mu]M) and caused an increase of cells in the S- and G2/M- phase (25, 50[mu]M) in HepG2 cells. High doses of Resv led to apoptosis (68.7[plusmn] 10.7%), phosphorylation of p53 (3-fold) and an increased acetylation of p53 (10-fold). It also reduced the intracellular NAMPT protein level by 48.1[plusmn] 15.4% and the NAMPT activity by 49.5[plusmn] 2.3%. Interestingly, the NAD level in HepG2 cells was not negatively modified. In contrast, primary human hepatocytes did not show significant changes in NAMPT protein expression and cell viability. However, primary human hepatocytes treated with the same amount of Resv showed a dose-dependently increased NAMPT activity (+112.0[plusmn] 7.5%).[br]Conclusion: We demonstrated that Resv exerts opposite effects on tumoral cells and primary, healthy hepatocytes regarding cell viability, apoptosis and NAMPT protein amount and -activity. The apoptotic effects of Resv on tumoral cells might be an up-regulation of the intracellular nicotinamide levels via reduced NAMPT activity and consequent inhibition of SIRT1 which leads to an increased p53 mediated apoptosis. The nicotinamide/NAD ratio could play a key role for sirtuin regulation.[br][br]Sources of Research Support: German Research Council - KFO 152 Atherobesity to W.K., the LIFE program (Leipzig Interdisciplinary Research Cluster of Genetic Factors, Clinical Phenotypes and Environment) to W.K., the IFB (Integrated Research and Treatment Center AdiposityDiseases), BMBF (Federal Ministry for Education and Research)to W.K.;the Kompetenznetz Adipositas-Verbund LARGE (TP01) and Kompetenznetz Diabetes to W.K.[br][br]Nothing to Disclose: SS, SP-Q, AG, MP, AB-O, SL, RG, WK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 476 58 382 SAT-280 PO42-01 Saturday 321 2012


322 ENDO12L_SAT-281 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) Tanshinone IIA Inhibits the Growth of Prostate Cancer Cell LNCaP Cells by the Transcriptional Activity of Androgen Receptor Ching-Yuan Wu, Chang-Yi Hsieh, Ko-En Huang, Chawnshang Chang, Hong-Yo Kang Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan; Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan; Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan Androgen receptor (AR) is a nuclear transcription factor of the hormone receptor family. For some prostate cancers, named AR-dependent prostate cancer, it plays a very important role for the development and progress of prostate cancer. In our study, we found tanshinone IIA can inhibit the growth of LNCaP cells, AR-dependent prostate cancer cell line. It suggests that tanshinone IIA can affect the function of AR. In our data, we found tanshinone IIA can profoundly inhibit the expression of mRNA and protein level in AR down-dream proteins. It suggests tanshinone IIA can block the transcriptional activity of activated ARs. Interestingly, tanshinone IIA also can suppress protein level and mRNA level of AR in our result. Hence, in this study, we discover tanshinone IIA can efficiency inhibit AR transactivation, block AR target genes expression, and inhibit cell growth in AR positive LNCaP cells. Our results suggest that tanshinone IIA could be a potential agent for prostate cancers.[br][br]Nothing to Disclose: C-YW, C-YH, K-EH, CC, H-YK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2376 58 383 SAT-281 PO42-01 Saturday 322 2012


323 ENDO12L_SAT-282 POSTER SESSION: Hormone Dependent Tumors (1:30 PM-3:30 PM) [sup]18[/sup]F-Fluorodihydroxyphenylalanine PET Imaging as Promising Diagnostic Tool for Breast Invasive Ductal Carcinoma Kathryn S King, Jaydira Del Rivero, Clara C Chen, Millie A Whatley, Karen T Adams, Karel Pacak National Institute of Child Health and Human Development, National Institute of Health, Bethesda, MD; National Institute of Health, Bethesda, MD [bold]Introduction: [/bold]Conventional imaging of breast cancer can be challenging due to the similarities in tissue density between normal breast parenchyma and cancerous tissue. We report two cases with pheochromocytoma and both were found to have invasive ductal carcinoma of the left breast that showed positive uptake with [sup]18[/sup]F-fluorodihyroxyphenylalanine ([sup]18[/sup]F-FDOPA). These cases introduce a possible new radiolabeled imaging agent for localizing and staging breast cancer lesions. [bold]Case: [/bold]First, a 51-year-old female with a right adrenal pheochromocytoma underwent surgical removal and pathological examination of the adrenal tumor verified the diagnosis. A variety of symptoms and her past history resulted in a recommendation for an MRI of the liver, on which a 1-2 cm nodule in the left breast was incidentally found. Subsequent imaging with MRI, mammogram, and [sup]18[/sup]F-fluorodeoxyglucose ([sup]18[/sup]F-FDG) positron emission tomography (PET)/computed tomography (CT) confirmed the presence of the left breast lesion. An [sup]18[/sup]F-FDOPA positively identified the left breast lesion, a core biopsy was performed that showed invasive ductal carcinoma. Second, a 49-year-old female with positive succinate dehydrogenase subunit B (SDHB) mutation with no biochemical evidence of pheochromocytoma/paraganglioma, was found on CT a small (8mm) adrenal mass. At the initial presentation the patient reported a left breast mass demonstrated on mammogram. A 1.7 cm mass in the left breast was seen on CT. Subsequent anatomical imaging with MRI and breast MRI confirmed the presence of this lesion. Functional imaging with [sup]18[/sup]F-FDG and [sup]18[/sup]F-FDOPA were positive for the left breast lesion. A biopsy was performed and reported invasive ductal carcinoma. [bold]Discussion:[/bold] [sup]18[/sup]F-FDOPA has not been evaluated to date in breast cancer, but amino acid transporters have been shown to be present on breast cancer cells. We presented two cases of invasive breast carcinoma that demonstrated positive [sup]18[/sup]F-FDOPA uptake. The potential use of [sup]18[/sup]F-FDOPA PET, or possibly positron emission mammography, in the localization of breast cancer lesions should be further studied. Additionally, the significance of amino acid receptors on these lesions should be investigated as they could be a possible avenue for treatment. The potential for this radiopharmaceutical as a diagnostic agent and the potential for it to provide insight into possible therapeutic targets are unknown and should be further studied.[br][br]Nothing to Disclose: KSK, JDR, CCC, MAW, KTA, KP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1348 58 384 SAT-282 PO42-01 Saturday 323 2012


324 ENDO12L_SAT-283 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) The Relationship of Papillary Thyroid Cancer to SDHB Paraganglioma Syndrome: Coincidental or Causal? Hussain Mahmud, Ruth E Dudley, Sue M Challinor, Linwah Yip, Michael T Stang, R Harsha Rao University of Pittsburgh Medical Center, Pittsburgh, PA; University of Pittsburgh Medical Center, Pittsburgh, PA; University of Pittsburgh Medical Center, Pittsburgh, PA BACKGROUND:[br]Mutations in the Succinate Dehydrogenase B (SDHB) gene are most commonly associated with abdominal paragangliomas (PGL) and less frequently with adrenal pheochromocytomas and head and neck paragangliomas (HNPGL). Isolated reports of coexisting papillary thyroid carcinoma (PTC) in patients with such mutations are of uncertain significance (1). We report three patients with PTC from two families with different SDHB mutations and a molecular pattern suggesting a sporadic, rather than genetic relationship.[br]CASES:[br]Patient 1: A 45 year old female with SDHB c.380T[gt]G mutation (found on genetic screening when her sister developed HNPGL) underwent a total thyroidectomy for bilateral nodules (left: 4.8 X 3.8 cm, right: 1.7 X 0.9 cm). A 0.9 cm follicular-variant PTC found in the right lobe was negative for BRAF, NRAS61, HRAS61, KRAS12/13 mutations or RET/PTC rearrangement. Loss of heterozygosity (LOH) for the SDHB gene was not observed.[br]Patient 2: A 40 year old male (sibling of Patient 1) with the same SDHB mutation and bilateral thyroid nodules (left: 1.4 X 1.2 cm, right: 1.2 X 1.1 cm) was found to have bilateral multifocal PTC (4 foci, 0.3 - 1.4 cm in size) on thyroidectomy, with the largest focus showing follicular variant PTC, positive for NRAS-61 mutation.[br]Patient 3: A 17 year old female with SDHB c.331_332delCT mutation (found on genetic screening as her brother had metastatic PGL) was found to have a left thyroid nodule (2.2 X 1.5 cm) on neck CT. FNA showed a follicular lesion with NRAS-61 mutation. She underwent thyroidectomy, which revealed a 2.1 cm encapsulated follicular variant PTC.[br]DISCUSSION:[br]Based on rare reports, PTC is included as a minor component of the SDHB-related PGL syndrome. However, earlier reports of PTC in this syndrome did not include tests for mutations and chromosomal rearrangements usually implicated in PTC pathogenesis. In one patient it was reported that there was no LOH in the PTC (2). In our series of three SDHB positive individuals with follicular-variant PTC, two had an NRAS61 mutation, a genetic signature of sporadic cancers. This suggests that SDHB is not involved in the pathogenesis of PTC and that its sporadic occurrence in these syndromes is likely to be coincidental. Based on this, we believe that PTC should not be considered as part of SDHB-related PGL syndromes, and that routine screening specifically for PTC (through thyroid ultrasonography) is unnecessary in such patients.[br][br](1) Neumann HP, et al. Distinct clinical features of paraganglioma syndromes associated with SDHB and SDHD gene mutations. JAMA. 2004 Aug 25;292(8):943-51. (2) Solis DC, et al. Penetrance and clinical consequences of a gross SDHB deletion in a large family. Clin Genet. 2009 Apr;75(4):354-63.[br][br]Nothing to Disclose: HM, RED, SMC, LY, MTS, RHR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1053 59 385 SAT-283 PO53-01 Saturday 324 2012


325 ENDO12L_SAT-284 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) The Association between Pheochromocytoma (Pheo)/Paraganglioma (PGL) and Cyanotic Congenital Heart Disease Aya Tsumagari, Akiyo Tanabe, Mika Tsuiki, Hirohumi Tomimatsu, Toshio Nakanishi, Atsuhiro Ichihara Institute of Clinical Endocrinology and Hypertension, Tokyo Women[apos]s Medical University, Tokyo, Japan; Tokyo Women[apos]s Medical University, Tokyo, Japan; Clinical Research Institute, Kyoto Medical Center, National Hospital Organization, Kyoto, Japan Cyanotic congenital heart disease and Pheo/PGL are rare disorders. Herein we report the clinical characteristics of 3 patients with Pheo/PGL associated with congenital heart disease. Case 1: As an infant, this 30-year-old man underwent palliative surgery for single-ventricle with right heart hypoplasia, transposition of the great arteries, and atrial septal defect. However, the patient had persistent cyanotic heart disease following the cardiac surgery. At 28 yrs of age, he presented with symptoms consistent with Pheo/PGL (eg, episodes of hyperhidrosis, palpitation, and paroxysmal hypertension). Biochemical testing included diagnostic elevations in plasma norepinephrine. Imaging detected a 5.5-cm PGL near the left renal vein. Case 2: At 2 months of age, this 13-year-old adolescent underwent palliative surgery for Tetralogy of Fallot. Cyanotic heart disease persisted postoperatively. At 10 yrs of age, she presented with paroxysmal hyperhidrosis; subsequently, at 13 yrs of age, the paroxysms included palpitations and fatigue. Blood pressure was normal. Biochemical testing included diagnostic elevations in plasma norepinephrine. Imaging detected a 4.4-cm right adrenal Pheo. Case 3: At 2 yrs of age, this 22-year-old woman underwent palliative surgery for Holt-Oram syndrome (complex cyanotic malformation of the heart, defect of the radius bone, and the anal atresia). At 20 yrs of age, she presented with hyperhidrosis. At 22 years of age she developed acute heart failure and hypertension. Biochemical testing included diagnostic elevations in plasma norepinephrine. Imaging detected a right peri-renal 2.3-cm PGL with associated hepatic and boney metastases. Due to their cyanotic heart disease and high surgical risk, these 3 patients were not surgical candidates. Medical treatment included alpha- and beta-adrenergic blockade, which resulted in symptomatic improvement. Although the pathophysiologic mechanisms responsible for the occurrence of Pheo/PGL in the setting of cyanotic congenital heart disease remain to be elucidated, it is known that the prevalence of PGL is increased in populations living at high altitude. In addition, succinate dehydrogenase subunit mutations are linked hypoxic signaling, predisposing to the development of Pheo/PGL. We speculate that the chronic hypoxia associated with cyanotic congenital heart disease may increase the risk of Pheo/PGL.[br][br]Nothing to Disclose: AT, AT, MT, HT, TN, AI 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 787 59 386 SAT-284 PO53-01 Saturday 325 2012


326 ENDO12L_SAT-285 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) A Case Report: Surgically Unresectable Metastatic Abdominal Paraganglioma Susan Michelle Tibuni-Sanders, Mary Ruppe The University of Texas Medical School at Houston, Houston, TX [bold]Introduction:[/bold][br]Malignant abdominal paragangliomas are rare neoplasms developing from extra adrenal chromaffin cells of sympathetic ganglia. These are rare causes of endocrine hypertension and can be a challenge for the clinician to treat when not surgically resectable. We now present such a case of malignant paraganglioma, and currently available management options.[br][bold]Clinical Case:[/bold][br]A 53 year old Hispanic man with a history of HTN and a family history of abdominal tumors presented to a university hospital complaining of cough, chest pain, and lower extremity edema. He was found to have a DVT, multiple pulmonary emboli, and non-ischemic CHF with an EF of 20%. CT imaging was performed which revealed a 4.5x3.7x8.5cm mass, anterior to the aorta, invading and compressing the right ureter, with multiple para-aortic lymph nodes.[br]Fine needle aspiration revealed a neuroendocrine neoplasm with positive staining for synaptophysin, neuron specific enolase, and chromogranin. MIBG scan showed uptake in abdominal lymph nodes. Plasma normetanephrines were 3385 pg/ml, N[lt]145. Plasma metanephrines were 101pg/ml, N[lt]62. 24 hour urine dopamine was 919 micrograms/24 hrs, N[lt]510. Chromogranin A was 48 nmol/L, N[lt]48.[br]During his hospital course his hypertension was difficult to control with severe orthostatic hypotension and episodic hypertensive spikes. The patient was treated with volume repletion and a high salt diet in order to allow for the titration of alpha and then beta blockers.[br]He underwent surgical exploration, but given tumor adherence to vital structures, the paraganglioma was incompletely resected. Pathology was consistent with a Stage IV T4N1M0 pheochromocytoma. He is now under consideration for chemotherapeutic options.[br][bold]Clinical Lessons:[/bold][br]As seen in this case, malignant paraganglioma represents a challenging clinical entity. Chemotherapy with established entities such as CVD (cyclophosphamide, vincristine, dacarbazine), MIBG, and other new therapies such as sunitinib have shown symptomatic and biochemical improvement, but incomplete tumor response. This case of unresectable paraganglioma highlights the need for a multidisciplinary endocrine and oncologic approach to achieve optimal patient outcomes.[br][br]Nothing to Disclose: SMT-S, MR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2215 59 387 SAT-285 PO53-01 Saturday 326 2012


327 ENDO12L_SAT-286 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Palliative Chemotherapy for Malignant Paraganglioma Masayasu Iwabuchi Seirei Mikatahara Hospital, Hamamatsu, Japan BACKGROUND[br]Malignant paraganglioma is rare, deceptive and deadly. Averbuch reported some success with the CVD therapy that consists of 750 mg/m2 of cyclophosphamide on day 1, 1.4 mg/m2 of vincristine on day 1, and 600 mg/m2 of dacarbazine on days 1 and 2 (1). Iwabuchi reported palliative chemotherapy for malignant pheochromocytoma (2). Prior to treatment, an informed consent should be obtained. In treating patient with psychiatric disorder, collaboration with psychiatrists is required.[br]CASE REPORT[br]A 52-year-old woman suffered from abdominal pain. CT scanning indicated a retroperitoneal lesion. Positive scanning of MIBG suggested that the tumor had arisen from the sympathetic ganglia. After surgical treatment, histological diagnosis of paraganglioma was confirmed and she was hospitalized in another mental institute because of schizophrenia. Next year, examinations of CT, MIBG and PET indicated metastases to rib in her chest wall and lumbar vertebrae. After the diagnosis of malignant paraganglioma was confirmed, she made a decision of chemotherapy under informed consent. She could share all medical information we had. We started the CVD chemotherapy with the collaboration of psychiatrists and other medical staffs. We have continued the treatment for 33 months until her psychiatric condition worsened. Because of delusion, she could not share any medical information. We decided to discontinue CVD therapy. Metastasized tumor turned worse during the discontinuation. She did not agree to re-start CVD therapy because of the side effects of the chemotherapy. She decided to have palliative chemotherapy of subcutaneous administration of 20 mg of cytosine arabinoside twice a week. The chemotherapy had been continued for eleven months without any side effect. However, her pregnancy delusion of schizophrenia obliged us to discontinue chemotherapy again. Three months later, we could re-start the chemotherapy because her psychiatric condition turned better enough to obtain informed consent again. Retrospectively, rapid worsening of metastasized lesion during discontinuation of the chemotherapy suggested the efficacy of the treatment.[br]RESULT[br]We treated a schizophrenic patient with malignant paraganglioma in collaboration with psychiatrists. The efficacy of palliative chemotherapy of low dose cytosine arabinoside was demonstrated retrospectively. The multidisciplinary discussion is prerequisite for the treatment of the patient with psychiatric disorder.[br][br](1) Averbuch et al., Ann Intern Med 1988; Aug 15;109(4):267-73. (2) Iwabuchi et al., Intern Med. 1999 May;38(5):433-5.[br][br]Nothing to Disclose: MI 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1169 59 388 SAT-286 PO53-01 Saturday 327 2012


328 ENDO12L_SAT-287 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Association of RET Haplotype, Not Linked to the RET C620R Activating Mutation, with Hirschsprung Disease in a MEN2 Family Rodrigo Almeida Toledo, Elisangela Quedas, Sergio Pereira de Almeida Toledo University of Sao Paulo School of Medicine, Sao Paulo, Brazil Background: It was initially thought that Hirschsprung disease could be due to disturbances in apoptosis or due to a tendency of the mutated RET receptor to be retained in the Golgi apparatus.[br]Objective: To report the results of the haplotyping analysis of a novel HSCR/FMTC family that carries the germline C620R RET activating mutation in exon 10.[br]Patients: The patients come from a rural area of northeast Brazil and have been diagnosed, treated and followed for several years at the University of Sao Paulo School of Medicine, Academic Hospital (Hospital das Clinicas, southeast Brazil).[br]Methods: To verify if an additional and inactivating RET mutation was present in the C620R-carrying HSCR patients in this family, the 21 exons of the RET proto-oncogene were analyzed, in duplicate, by polymerase chain reaction amplification and bidirectional sequencing, as previously reported.[br]Results: No germline RET mutation other than C620R was observed. We also investigated whether 16 RET single nucleotide polymorphisms (SNPs) or specific haplotypes were involved. The haplotype CTCGTGGTGAdel was documented in all of the studied C620R-mutated relatives in the present family, indicating that this haplotype is linked to the RET mutation. Because not all of family members with the RET mutation had developed HSCR, the emerging hypothesis was that HSCR disease may be associated with the remaining chromosomal region. In accordance, the MEN2-affected relatives who did not have HSCR have inherited the CATACTGTAAdel haplotype, while the HSCR/MEN2 patients had inherited a different haplotype, TTCGTTAAAGC. Furthermore, analysis of four additional members of this family with the C620R mutation (three cases without and one with HSCR) strengthened the proposal that this RET gene region is involved in the development of HSCR (n = 9; p = 0.012; Fisher exact test). Importantly, RET exon 2 A45A SNP has been found in the homozygous state in cases with HSCR only, whereas MEN2 patients and healthy controls are frequently heterozygous for this SNP.[br]Discussion: In summary, we have presented new evidence supporting the role of a RET haplotype, which is not linked to the C620R mutation, in the development of HSCR in a newly reported FMTC/MEN2 family.[br][br]Sources of Research Support: The study was funded by the CNPq research grant number 401990/2010 - 9.[br][br]Nothing to Disclose: RAT, EQ, SPdAT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1332 59 389 SAT-287 PO53-01 Saturday 328 2012


329 ENDO12L_SAT-288 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Bone Pain: An Atypical Presentation of Malignant Pheochromocytoma Maria Elena Pena, Ageliki Valsamis, Isabela Romao, John Tloczkowski North Shore University Hospital-LIJ, Manhasset, NY [bold][underline]Introduction[/underline][/bold]: Pheochromcytomas are rare and malignant pheochromocytomas are even less common. Both benign and malignant pheochromocytomas present similarly with the classic triad of palpitations, diaphoresis and headaches. Paroxysmal hypertension, orthostatic hypotension, and pallor are also common manifestations of pheochromocytomas (1).[br][bold][underline]Case[/underline][/bold]: A 32 year old Caucasian male presented to NSUH-LIJ complaining of localized left sided chest and back pain. He had been feeling well except for occasional palpitations and nausea. In the hospital, he was tachycardic with labile blood pressure. His course was complicated by a non ST-elevation myocardial infarction and respiratory distress requiring intubation. A CT scan revealed multiple pulmonary nodules, multiple lytic lesions in the left ribs and scapula and a 19 cm adrenal mass with necrosis. Plasma metanephrines were elevated: normetanephrines 397 nmol/L ([lt]0.90) and metanephrines 8.4 nmol/L ([lt]0.50). A 24-hour urine collection for catecholamines was markedly elevated consistent with a pheochromocytoma [norepinephrine 1451 mcg (15-80), epinephrine 267 mcg (0-20), dopamine 1562 mcg (0-20)]. Both aldosterone (45 ng/dL) and renin (27 ng/mL/h) were elevated suggesting secondary hyperaldosteronism possibly due to vascular compression. The patient[apos]s blood pressure was regulated with alpha blockade followed by beta blockade. Biopsy of a rib lesion was consistent with metastatic pheochromocytoma. Once stabilized, he underwent palliative debulking surgery. Review of tumor histology showed mitosis and nuclear pleomorphism with areas of necrosis, capsular invasion, positive synaptophysin staining, and low to absent S-100 staining. These features were consistent with malignant pheochromocytoma. The patient received chemotherapy with cyclophosphamide, vincristine, and dacarbazine but he did not respond to it. Targeted radiation therapy to the lytic lesions was attempted but he did not tolerate it. He is currently receiving supportive care.[br][bold][underline]Discussion[/underline][/bold]: Malignant pheochromocytoma presenting as bony pain is a rare phenomenon only described in a few case reports. He had few of the classic symptoms associated with pheochromocytoma despite having an adrenal mass that measured 19 cm. The symptoms of pheochromocytoma may be subtle at times so it is important to keep a high index of suspicion, especially in patients with hypertension.[br][br]Wailly P et al.,Langenbecks Arch Surg 2011; 239[ndash]246.[br][br]Nothing to Disclose: MEP, AV, IR, JT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1555 59 390 SAT-288 PO53-01 Saturday 329 2012


330 ENDO12L_SAT-289 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) A Patient with PTEN Hamartoma Tumor Syndrome (PHTS) and Multiple Pulmonary Tumorlets and Carcinoids Tobias Else, Victoria M Raymond, Jessica C Long, Gary D Hammer University of Michigan, Ann Arbor, MI Introduction: Roughly 80% of patients with the clinical diagnosis of Cowden syndrome are found to have PTEN mutations, which defines the diagnosis PTEN Hamartoma Tumor Syndrome (PHTS). PHTS predisposes to several cancers, including breast, endometrial, thyroid and renal cancers, as well as gastrointestinal polyposis. Foregut neuroendocrine tumors can be associated with von Hippel-Lindau disease, tuberous sclerosis and multiple endocrine neoplasia type 1 (MEN1), of which only MEN1 predisposes to pulmonary neuroendocrine tumors.[br]Clinical case: A 63-year old female patient presented for evaluation of multiple lung nodules. The patient had a history of thyroid cancer and breast cancer, treated with thyroidectomy and bilateral mastectomy in her 20s. She also underwent hysterectomy and bilateral salpingo-oophorectomy for unknown tumor pathology in her 30s. Her family history was notable for brain cancer in one first-degree relative. On physical exam, classical findings of Cowden syndrome were evident, including head circumference of 62cm, acral hyperkeratotic lesions and mucosal cobblestoning. Lung imaging showed multiple lung nodules ranging from 0.1 to 1.2cm. Diagnostic wedge resection revealed multiple tumorlets and typical carcinoids with absent mitosis. Sequencing of the PTEN gene revealed a mutation generating a premature stop codon (c.388C[gt]T, p.R130X). Hormonal work-up, including ACTH, cortisol, 5HIAA, gastrin, calcitonin, pancreatic polypeptide, chromogranin A, and metanephrines, was negative. The diagnosis of multiple non-functional neuroendocrine tumors of the lung (tumorlets with at least two dominant typical carcinoids) was made. Initial six month repeat imaging showed stable pulmonary nodules. In the absence of symptoms, the decision to follow the patient with regular clinic visits and imaging.[br]Conclusion: This is the first report on pulmonary neuroendocrine tumorlets in a patient with PHTS, possibly extending the tumor spectrum associated with this syndrome. Tumorlets have not been reported with other hereditary cancer syndromes. However, patients with tumorlets may have a higher incidence of non-neuroendocrine primary malignancies. Tumorlets, even with dominant nodules greater than 5mm, often follow a long indolent course of disease and can be followed by imaging.[br][br]Sources of Research Support: NIH Grant T32-DK007245.[br][br]Nothing to Disclose: TE, VMR, JCL, GDH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1422 59 391 SAT-289 PO53-01 Saturday 330 2012


331 ENDO12L_SAT-290 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Atrophic Gastritis, Hypergastrinemia and Multiple Gastrointestinal Carcinoids Associated with a Pituitary Macroadenoma Alejandro G Trepp Carrasco, Robert J Anderson Creighton University Medical Center, Omaha, NE [bold]Introduction:[/bold] The development of gastric carcinoid tumors in the setting of hypergastrinemia secondary to atrophic gastritis is well known. However, in the absence of documented MEN1, there is only one case, to our knowledge, of an additional association with a pituitary adenoma (GH hormone-producing).(1)[br][bold]Case:[/bold] Our patient is a 53 yo woman with a history of multiple autoimmune disorders including vitiligo and myasthenia gravis during childhood. At age 28, she had worsening Graves[apos] orbitopathy with hyperthyroidism and received radioiodine. By age 35, an EGD done for iron deficiency anemia revealed atrophic gastritis and a 4mm gastric carcinoid tumor with negative gastrin stain. At age 45, an MRI of the brain for headache showed a 12mm pituitary mass, which has remained stable in size. Her laboratory tests included positive IF antibodies and persistent hypergastrinemia (1500-6000pg/mL). Chromogranin A has been mildly increased ([lt] 300ng/mL); urinary 5-HIAA, catecholamines and metanephrines have been normal. Pituitary functions included: elevated postmenopausal LH and FSH; prolactin, 10-25ng/mL; variable TSH; normal ACTH and cortisol. IGF-1 has been high normal to slightly increased (up to 339ng/mL), and GH suppressed normally during OGTT. Calcium, PTH and vitamin D have remained normal. No pathogenic mutations were detected in the MEN1 gene. Multiple CT scans of the trunk and two octreotide scans have been negative. After multiple EGDs, two additional gastric carcinoid polyps (with negative gastrin stain) have been found. Autoimmune metaplastic atrophic gastritis also has been a persistent finding. Recently, a 4mm duodenal submucosal carcinoid polyp was found (positive gastrin stain).[br][bold]Conclusions:[/bold] In the setting of a negative MEN1 mutation test, this is the first report, to our knowledge, of a nonfunctional pituitary macroadenoma associated with Type A atrophic gastritis and multiple indolent, GI carcinoids, potentially caused by the hyperplastic/neoplastic effect of chronic hypergastrinemia. This effect possibly could have contributed to the pituitary adenoma formation as well.(1) Even though not 100% sensitive, MEN1 gene testing and other investigations for MEN1 are reasonable to do in similar patients. After 18 years of follow-up, we confirm a good prognosis for type I sub-centimeter gastric carcinoid tumors without metastases, and our findings support a conservative approach with periodic surveillance and endoscopic resection of polypoid lesions.[br][br](1) Kojima K et al., Intern Med. 1997;36:787[ndash]9.[br][br]Nothing to Disclose: AGTC, RJA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 870 59 392 SAT-290 PO53-01 Saturday 331 2012


332 ENDO12L_SAT-291 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Pasireotide and Octreotide LAR Administered in Combination in a Patient with Small Intestinal Neuroendocrine Tumor with Hypokalemia and a High Level of Vasoactive Intestinal Peptide Gordana Kozlovacki, Staffan Welin, Barbro Eriksson, Dan Granberg, Kjell Oberg, Eva Tiensuu Janson Uppsala University Hospital, Uppsala, Sweden Introduction: Small intestinal neuroendocrine tumors (NET) are argentaffin-positive tumors that progress slowly. We present a case of small intestinal NET secreting high levels of vasoactive intestinal peptide (VIP) causing severe diarrhea and hypokalemia that showed a good clinical and biochemical response to combination somatostatin analog therapy, including pasireotide and octreotide LAR.[br]Materials and methods: A 70-year-old woman who underwent surgery for small intestinal NET in 1991 had slowly deteriorating health during the spring of 2008, with increasing bouts of diarrhea followed by hypokalemia. High levels of circulating VIP were detected, though urinary 5-HIAA levels were normal.[br]Results: The patient started treatment with pasireotide 2400 micrograms/day in April 2008, and octreotide LAR was added in October 2008 at a dose of 30 mg every fourth week. This combination resulted in decreased episodes of diarrhea, from 10 to 4 per day, and the patient[apos]s need for potassium substitution diminished. The VIP level decreased from 788 pmol/L in October 2008 to the lowest level of 79 pmol/L in October 2010.[br]Conclusion: We present a rare case of small intestinal NET producing a high level of VIP. Combination therapy with pasireotide and octreotide LAR proved to be an effective treatment that resulted in decreasing episodes of diarrhea and decreasing secretions of VIP.[br][br]Sources of Research Support: Novartis Pharmaceuticals Corporation.[br][br]Disclosures: KO: Advisory Group Member, Novartis Pharmaceuticals, Pfizer, Inc., Ipsen. Nothing to Disclose: GK, SW, BE, DG, ETJ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1767 59 393 SAT-291 PO53-01 Saturday 332 2012


333 ENDO12L_SAT-292 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Refractory Hypoglycemia Post Octerotide Treatment of Recurrent Neuroendocrine Tumor Eli Klam, Mohamad Lazkani, Adonis Jabbour, Mohamad H Horani Banner Baywood Hospital, Mesa, AZ; Banner Good Samaritan Hospital, Phoenix, AZ; Alsham Endocrinology, Chandler, AZ [bold]Introduction[/bold][br]In this case study, we present a female with recurrent neuroendocrine pancreatic cancer that developed into an insulinoma. Aggravation of hypoglycemia in insulinoma by the long acting Somatostatin analogues is reported in this case.[br][bold]Case Report[/bold][br]An 84-year-old female had a history of treated breast cancer and a low grade pancreatic carcinoid tumor status post resection of most of the pancreas and splenectomy 14 years ago. She had no hypoglycemia at that time.[br]The patient had been experiencing flushing and carcinoid symptoms. An outside imaging revealed an 8.7 perihepatic mass and 3.5 cm left adrenal mass. Perihepatic mass was biopsied, and found to be a pancreatic endocrine neoplasm. She was treated with sandostatin for carcinoid symptoms. Two weeks later, she presented with near-syncope, cold sweats, and blood glucose of 39. Blood drawn during a hypoglycemic episode showed an unexpectedly normal insulin level and a high C-peptide level, confirming the insulinoma. MEN-I was ruled out. The adrenal mass was nonfunctional. Octreotide scan showed an intense tracer only in the pancreas consistent with neuroendocrine tumors. Dextrose drip was initially used to treat her hypoglycemia. Blood glucose levels were later maintained within normal ranges with use 10 mg dose prednisone. The insulinoma was not amenable to surgical resection. Patient was started on Everolimus and remained euglycemic on follow up blood glucose levels after 2 months from discharge.[br][bold]Discussion:[/bold][br]Pancreatic neuroendocrine tumors are rare and found in approximately 4 people per million in a year. Somatostatin and its long-acting analogues are effective in controlling symptoms in patients with functional neuroendocrine tumors. Somatostatin analogues however can aggravate hypoglycemia in patients with insulinoma (1). Using prednisone to maintain normal blood glucose levels after stopping Somatostatin analogues was rarely reported (2). The treatment of choice of insulinoma is surgical removal. Everolimus has been shown to induce rapid plasma glucose normalization in insulinoma within 14 days (3).[br][br]1. Acta Endocrinol (Copenh). 1989 Jul;121(1):34-40. 2. Support Care Cancer. 2005 Sep;13(9):760-2. 3.Oncologist. 2011;16(6):783-7.[br][br]Nothing to Disclose: EK, ML, AJ, MHH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 29 59 394 SAT-292 PO53-01 Saturday 333 2012


334 ENDO12L_SAT-293 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) A Case of ACTH-Secreting Bronchial Carcinoid Donato Iacovazzo, Francesca Lugli, Antonio Bianchi, Antonella Giampietro, Sabrina Chiloiro, Vincenzo Cimino, Mariella Celico, Giuseppe Grande, Germano Perotti, Giorgio Treglia, Vittoria Rufini, Pierluigi Granone, Alfredo Pontecorvi, Laura De Marinis Policlinico [quot]A Gemelli[quot] - Catholic University, Rome, Italy; Policlinico [quot]A Gemelli[quot] - Catholic University, Rome, Italy; Policlinico [quot]A Gemelli[quot] - Catholic University, Rome, Italy Ectopic ACTH syndrome (EAS) occurs in about 5-10% of ACTH-dependent hypercortisolism and, in up to 15% of cases, is related to occult neoplasms.[br]We report one case of EAS highlighting the usefulness of Nuclear Medicine imaging in pre and intraoperative localization of the ACTH-secreting neoplasm.[br]A 66 years old male patient was diagnosed in 2010 with ACTH-dependent hypercortisolism. Pituitary MRI was negative, while a chest-abdomen CT scan described a 7 mm nodule in the upper lobe of the right lung. Bilateral Inferior Petrosal Sinus Sampling (BIPSS) showed no evidence of center-periphery ACTH gradient. An Octreoscan did not reveal any area of pathological hyperuptake. Treatment with ketoconazole and lanreotide was started with clinical benefit. In July 2011 the patient underwent a PET/CT scan with 68Ga-DOTANOC that revealed mild hyperuptake of the lung nodule.[br]In September 2011 the patient underwent radio-guided surgery using a hand-held gamma probe 24 h after i.v. administration of 111In-pentetreotide. The pulmonary nodule was identified, confirmed by the gamma probe and surgically removed. Pathology confirmed a typical bronchial carcinoid with positive ACTH expression.[br]This case underlines how the integration of traditional diagnostic tools with both Nuclear Medicine imaging and intraoperative techniques can guarantee an improvement in the diagnostic and therapeutic management of neuroendocrine neoplasms.[br][br]Nothing to Disclose: DI, FL, AB, AG, SC, VC, MC, GG, GP, GT, VR, PG, AP, LDM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2056 59 395 SAT-293 PO53-01 Saturday 334 2012


335 ENDO12L_SAT-294 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Midgut ACTH-Secreting Neuroendocrine Tumor Unmasked with FDOPA-PET Julien Ducry, Fulgencio Gomez, John O Prior, Ariane Boubaker, Matteo Monti, Maurice Matter, Braulio Mora, Nelly Pitteloud, Luc Portmann Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland; Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland; Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland; Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland; Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland [bold]Introduction[/bold]: Ectopic ACTH syndrome (EAS) accounts for 5-10% of all cases of Cushing syndrome, mainly caused by neuroendocrine tumors (NETs) of lungs, pancreas, thymus, and other less frequent locations. Once the non-pituitary origin of excess ACTH has been established, CT scan, MRI, somatostatin receptor scintigraphy (SSRS), and [sup]18[/sup]F-deoxyglucose-PET (FDG-PET) are used to localize secreting tumors, but owing to small tumor size and tissue diversity, the task can be challenging.[br][bold]Clinical case[/bold]: A 64-year old man presented with fatigue, muscular weakness, and dyspnea, rapidly developing in four months. On examination, he was in poor physical condition, and showed slight facial redness, marked proximal amyotrophy, and bruises. Laboratory analyses revealed hypokalemia (1.7 mmol/L) with metabolic alkalosis, elevated random serum cortisol (1600 nmol/L; 5PM, N: 40-250), ACTH (261 pg/L; 5PM, N: 5-35), and urinary free cortisol (9181 nmol/d; N: 20-220). During a 48-hour oral dexamethasone test (2 mg q6h), serum cortisol decreased from 1221 to 721 nmol/L. Pituitary MRI was normal, bilateral inferior petrosal sinus sampling, with CRH stimulation, showed no significant central-to-periphery ACTH gradient. Therefore EAS was diagnosed. Neck, thorax and abdomen CT scan, MRI, SSRS, and FDG-PET failed to localize any lesion. Finally, [sup]18[/sup]F-dihydroxyphenylalanine-PET (FDOPA-PET), coupled with CT scan disclosed a 20-mm lesion in the jejunum wall and a 7-mm lymph node. During surgery, a segment of mid-jejunum was resected showing two adjacent NETs, measuring 2 cm (Ki 67 1%)(T4), and 0.5 cm (Ki 67 1%)(T2), with a local peritoneal deposit (M1). On immuno-histochemistry, the largest tumor expressed ACTH in 30% of cells. After surgery ACTH and cortisol fell, the patient fully recovered, after transient adrenal insufficiency, with a normal cortisol on a follow-up of 5 months.[br][bold]Discussion[/bold]: Midgut NETs causing EAS are difficult to localize on CT or MRI, and require from metabolic imaging. However, SSRS may be negative in presence of elevated cortisol due to a down regulation of type-2 somatostatin receptors. NETs are often poor candidates for FDG-PET due to their low mitotic activity. FDOPA-PET, related to amine precursor uptake, has been shown to have a low sensitivity for occult EAS (55%), but an excellent positive predictive value (100%), and therefore may constitute a useful alternative tool for occult EAS when all the other conventional radiological and nuclear medicine have failed.[br][br]Nothing to Disclose: JD, FG, JOP, AB, MM, MM, BM, NP, LP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 915 59 396 SAT-294 PO53-01 Saturday 335 2012


336 ENDO12L_SAT-295 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Insulinoma with Non-Suppressed Ketones at the Completion of a 72-Hour Fast Abdulrahman Alkabbani, Robert Zimmerman Endocrinology and Metabolism, Cleveland, OH [bold]Introduction[/bold]:[br]The most recent endocrine society clinical practice guidelines for adult hypoglycemia indicate that betahydroxybutyrate (BHB) levels do not exceed 2.7 mmol/L in patients with insulinoma undergoing a 72 hour fast test, based on a paper by O[apos]Brien et al. showing a single outlier with this value in their series. We hereby present a case of pathologically proven insulinoma with progressive elevation of BHB well beyond the cut off level on a 72-hr fast.[br][bold]Clinical Case:[/bold][br]An 86-year old woman was referred with frequent episodes of fatigue, sweating and confusion for a year. The episodes occurred during fasting and resolve by eating. Her family reported 2 occasions of loss of consciousness with corresponding capillary blood glucose of 48 and 27 mg/dL. No weight change was noted. A 72-h fast test demonstrated a spontaneous and symptomatic drop in serum glucose to [lt]55 mg/dL after 15 hrs. Glucose (65-100 mg/dL), insulin (1-24 uU/mL), proinsulin (3-20 pmol/L) [amp] c-peptide (0.8-3.2 ng/mL) levels were (45, 2.2, 28, 0.7) at 15 hrs, (43, 3.2, 34, 0.7) at 18 hrs, (39, 3, 35,-) at 20 hrs, (33, 6.2, 44, 0.9) at 22 hrs of fasting, respectively. BHB (0.0-0.3 mmol/L) was progressively rising during the fast to 0.8, 1.3, 2.6, and 3.5 mmol/L at the same intervals respectively.[br]The test was concluded by administrating 1 mg glucagon IV which showed an increase in glucose from 32 to 73 mg/dL after 30 minutes. Insulin antibodies and serum sulfonylurea screening were negative. Lightheadedness, confusion and sweating resolved after feeding.[br]CT of the abdomen didn[apos]t show a pancreatic mass, so the patient underwent selective arterial calcium stimulation test which showed a step-up in insulin level at the mid pancreatic region. Intraoperative ultrasonography localized a 1 cm tumor at the tail of the pancreas which was enucleated, and pathology confirmed a 0.9 cm pancreatic endocrine neoplasm, consistent with insulinoma.[br]The patient had an uneventful recovery after surgery with resolution of hypoglycemia.[br][bold]Conclusion:[/bold][br]Despite the antiketogenic effect of insulin and consequently the expected suppression of BHB levels in fasted insulinoma patients, our patient demonstrated progressive BHB elevation with a peak well beyond the cutoff value established in recent guidelines. Therefore, in cases with high clinical suspicion for insulinoma, rising BHB levels should not be a reason for early termination of the fast before establishing the diagnosis using the other diagnostic parameters.[br][br]O[apos]Brien T et al.,J Clin Endocrinol Metab. 1993 Aug;77(2):448-51.[br][br]Nothing to Disclose: AA, RZ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2032 59 397 SAT-295 PO53-01 Saturday 336 2012


337 ENDO12L_SAT-296 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Adrenocortical Carcinoma (ACC) Associated with Birt-Hogg-Dub[eacute] Syndrome Tobias Else, Victoria M Raymond, Jessica C Long, Elena M Stoffel, Hammer D Gary University of Michigan, Ann Arbor, MI Introduction: Adrenocortical carcinoma (ACC) has been described as part of several familial cancer susceptibility syndromes, including Li-Fraumeni syndrome, multiple endocrine neopasia type 1, and Beckwith-Wiedemann syndrome. Birt-Hogg-Dub[eacute] syndrome (BHD) is a rare familial cancer susceptibility syndrome characterized by the dermatological triad of trichodiscomas, fibrofolliculomas and acrochordons. BHD predisposes to the development of benign and malignant kidney tumors (specifically renal oncocytomas) and pulmonary cysts associated with spontaneous pneumothoraces.[br]Clinical case: A 62-year old female patient was diagnosed with a trichodiscoma of the left cheek. Because trichodiscomas are classically associated with BHD and otherwise rare in the general population, suspicion for the diagnosis of BHD was raised. Baseline renal imaging revealed a 6.0x5.0cm adrenal tumor. The patient underwent a surgical resection, and pathology was notable for a low grade ACC, oncocytoma type. Germline sequencing of the folliculin gene (FLCN) revealed a deleterious frameshift mutation resulting in a premature stop codon (c.1252delC, p.Leu418TrpfsX50), previously reported in patients with BHD. Sequencing and dosage analysis of TP53 responsible for Li-Fraumeni Syndrome was negative. In order to further characterize the adrenal phenotype of BHD patients, we evaluated the adrenal morphology in patients with BHD and deleterious FLCN mutations who enrolled in the IRB-approved University of Michigan Cancer Genetics Registry (n=10). Abdominal imaging (CT or MRI) was available for all patients and did not reveal any gross adrenal abnormalities. None of the patients reported adrenal tumors in their families.[br]Conclusion: This is the first description of a patient with BHD and ACC. Of note, a low grade, oncocytic ACC is a highly unusual tumor. Benign oncocytomas as well as malignant clear cell/oncocytic hybrid renal tumors are classically found in BHD patients. In conclusion, our data suggest ACC, specifically adrenal oncocytoma, as a rare feature of the BHD tumor spectrum.[br][br]Sources of Research Support: NIH Grant T32-DK007245.[br][br]Nothing to Disclose: TE, VMR, JCL, EMS, HDG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1853 59 398 SAT-296 PO53-01 Saturday 337 2012


338 ENDO12L_SAT-297 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Adrenocortical Carcinoma: An Unusual Case Joana Menezes, Elisabete Rodrigues, Francisco Monteiro, Joanne Lopes, Jose Costa Maia, Davide Carvalho Centro Hospitalar S[atilde]o Jo[atilde]o, Porto, Portugal; Centro Hospitalar S[atilde]o Jo[atilde]o, Porto, Portugal; Centro Hospitalar S[atilde]o Jo[atilde]o, Porto, Portugal AIM: To describe a successful long-term tumor control in metastatic adrenocortical carcinoma, a relatively rare tumor with poor prognosis.[br]CASE REPORT: A 41-year-old woman with medical history notable just for hypertension initially presented in January 2007 in a private practice hospital with right lumbar back pain, peripheral oedema, virilization, amenorrhea and weight gain (8Kgs in 3 months). On physical examination she presented with Cushing[apos]s stigma. Endocrine tests confirmed high cortisol levels and excess levels of steroid hormone (total testosterone, delta(4)-androstenedione, dihydroepiandrostenedione sulfate and 17-hydroxyprogesterone) with normal levels of aldosterona, renina and catecholamines. CT scan revealed an adrenal mass measuring 9 x 7.5 x 8 cm with a mass effect on surrounding structures (liver, right kidney and inferior vena cava) but without invasion. The patient underwent laparoscopic surgery in April 2007 and histology revealed an adrenocortical carcinoma (pT2N0M0, stage II, ENSAT 2008). She started therapy on corticosteroids with progressive dose-reduction and clinical and laboratory tests resolution. Two years later, in May 2009, a follow-up CT scan revealed a 3cm liver metastasis and a biopsy confirmed metastasis of primary adrenocortical carcinoma with mitotic index 1m/10HPF. She underwent partial hepatectomy and was referred to our Department but missed several consultations and attended our first only in June 2011. In July 2011 the follow-up CT scan demonstrated suspicious implant of 6mm on the right inferior angle of the liver and we performed a fluorodeoxyglucose (FDG)-PET scan that showed no pathologic findings.[br]CONCLUSION: Adrenocortical carcinoma is an uncommon and aggressive malignancy with approximately 30-85% of patients having distant metastasis at the time of presentation(1,2). We present an unusual case of adrenocortical carcinoma non metastatic on diagnostic time with low rate of progression despite not being on mitotane therapy, with 4.5 years survival so far.[br][br](1)Martin Fassnacht et al, Nat Rev ENdocrinol 2011; 7:323. (2)Martin Fassnacht et al, Clinical Endocrinol 2919; 73:565.[br][br]Nothing to Disclose: JM, ER, FM, JL, JCM, DC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1428 59 399 SAT-297 PO53-01 Saturday 338 2012


339 ENDO12L_SAT-298 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Von Hippel-Lindau Disease with Pancreatic Neuroendocrine Tumor Causing Ectopic Cushing Syndrome Esra Hatipoglu, Hasan Kepic, Elif Rusen, Levent Kabasakal, Sadi Gundogdu, Pinar Kadioglu Cerrahpasa Medical School, University of Istanbul, Istanbul, Turkey; Cerrahpasa Medical School, University of Istanbul, Istanbul, Turkey Introduction: Von Hippel Lindau Disease(VHLD)is an autosomal dominant disease characterized by various tumors.Pancreatic neuroendocrine tumor(PNET)is a rare presentation of VHLD.In VHLD PNETs are usually indolent.[br]Case: A 39 years old woman with previous diagnosis of VHLD presented with Cushing[apos]s syndrome (CS) persisting after unilateral adrenalectomy for pheochromocytoma. Twentyfour hour urine valine mandelic acid, homovalinic acid, metanephrine and normetanephrine levels were 0.54 mg/24 hr,4.07 mcg/24 hr,131.67 mcg/24 hr and 94.61 mcg/24 hr, respectively (n[lt]6.6 mg/24 hr, [lt]6.9 mcg/24 hr,[lt]297 mcg/24 hr and [lt]354 mcg/24hr,respectively). Initial tests were consistent with ACTH dependent CS: urinary free cortisol level [gt]1000 nmol/24 hr (n[lt] 100 nmol/24 hr), cortisol after 1 mg dexamethasone overnight test-21 mcg/dl (n[lt] 1.8 mcg/dl), after high dose dexamethasone test cortisol level-19.4 mcg/dl (n[lt] 1.8 mcg/dl), ACTH level-68.40 pg/ml (n=0-46 pg/ml). Nonsupressed cortisol level after 8 mg dexamethasone overnight test,normal sella MRI,basal ACTH level with nonsupressed DHEA-SO4level (264.2 mcg/dl, n=60.8-338 mcg/dl) aroused the suspicion of ectopic CS. Abdominal CT revealed a 13 mm noncalcified nodule in the left lung and pancreatic lesions which showed rapid contrast enhancement in arterial phases. Galium-68 Somatostatin Receptor PET revealed a primary tumor with intense somatostatin type 2 receptor involvement in the pancreatic head with metastatic lesions in aortocaval lymphatic system and in the left lung.With endoscopic ultrasonography fine needle aspiration biopsy was performed to primary tumor. The cytologic and immunocytochemical findings were consistent with a low grade neuroendocrine tumor (synaptophysin and chromogranin were strongly positive, CD56 was focal membranous positive and Ki67 index was below 1%). Because the tumor was metastatic peptide receptor radionuclide therapy with [177Lutetium-DOTA0,Tyr3]octreotate for a total of 4 cycles once in every 6-8 weeks was planned.She did not live any side-effects and was discharged from the hospital 10 days after the first cycle.[br]Conclusion: Timely diagnosis of PNETs in VHLD is of clinical importance. Although they are notorious for being indolent they may have a wide spectrum of hormonal activity and may be diagnosed when they are metastatic. So accurate diagnosis warrants a high index of suspicion and a multidisciplinary approach. Treatment should be individualized based on patient and tumor characteristics.[br][br]Nothing to Disclose: EH, HK, ER, LK, SG, PK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 757 59 400 SAT-298 PO53-01 Saturday 339 2012


340 ENDO12L_SAT-299 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Pancreatic Neuroendocrine Tumor Presenting with New-Onset Diabetes Mellitus and Multiple Endocrinopathies: Rare Occurrence Visha Amit Parnerkar, Crispin Semakula Hennepin County Medical Center, Minneapolis, MN A 42 year old female with history of hypertension presented with polyuria, polydipsia and hyperpigmentation. She was newly diagnosed with uncontrolled diabetes mellitus (DM)(HgbA1C 15.6) requiring three oral hypoglycemic agents and over 200 Units of insulin daily. Physical examination showed normal blood pressure, central obesity, violaceous abdominal stria, hyperpigmentation on palmer creases, trunk and extremities.[br]Biochemical results were remarkable for Gastrin [gt]10,000 pg/ml (0-100 pg/ml), elevated morning Cortisol (43.1), ACTH 441pg/ml (7.2-63.3 pg/ml), low CRH ([lt]28pg/ml) and hypokalemia (K 2.9 mEq/L). Patient had elevated cortisol levels on both low and high dose dexamethasone suppression tests confirming the diagnosis of ectopic ACTH production consistent with ACTHoma. DM was secondary to Cushing[apos]s syndrome. Despite high gastrin levels consistent with Gastrinoma, there was no evidence of Zollinger- Ellison syndrome clinically or on upper GI endoscopy.[br]CT Abdomen showed a mass (12 X11X10 cm) at the tail of pancreas with liver nodules. Ultrasound guided biopsy reaveled neuroendocrine tumor with immunohistochemistry stains positive for pankeratin, synaptophysin and chromogranin. Octreotide scan showed multiple metastatic lesions in the liver.[br]She underwent transcatheter arterial chemoembolization of the hepatic artery for liver metastases. Initial treatment with octreotide and ketoconazole failed. So, debulking surgery was performed which restored ACTH, cortisol levels and euglycemia (HgbA1c 4.5). Patient was off insulin in next two months. Coincidentally, she developed severe acute hypercalcemia presenting with altered mental status secondary to adrenal insufficiency. Other results showed low aldosterone, hypovitaminosis D, normal calcitonin, PTHrp and 24-hour urine calcium excretion. There was no bone metastases on radiological studies. She responded well to fluid resuscitation and hydrocortisone. Discharge plan included slow long-term taper of steroid replacement.[br]Pancreatic neuroendocrine tumors may secrete active hormones resulting in clinical syndromes. Most common are Insulinomas (60%) and gastrinomas (20%). ACTHomas (0.8%), VIPOmas, Glucagonomas are rare. Gastrinomas are derived from multipotent cells of neuroendocrine origin. They are also a part of MEN syndrome. This case illustrates the rare occurrence of pancreatic neuroendocrine tumor producing two ectopic active hormones with major manifestations from ACTH overproduction.[br][br]1.Pancreatic neuroendocrine tumors (PNETs): incidence, prognosis and recent trend toward improved survival. Halfdanarson TR, Rabe KG, Rubin J, Petersen GM. Ann Oncol. 2008;19(10):1727. 2. Clinicopathological features of pancreatic endocrine tumors: a prospective multicenter study in Italy of 297 sporadic cases. Zerbi A, Falconi M, Rindi G, Delle Fave G, Tomassetti P, Pasquali C, Capitanio V, Boninsegna L, Di Carlo V, AISP-Network Study Group. Am J Gastroenterol. 2010;105(6):1421. 3. Molecular genetics of neuroendocrine tumors.Duerr EM, Chung DC.Best Pract Res Clin Endocrinol Metab. 2007;21(1):1. 4. Pancreatic neuroendocrine tumors. H.L.O[apos]gary, K.C.Conlon.EJSO 34 (2008) 324-332.[br][br]Nothing to Disclose: VAP, CS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 713 59 401 SAT-299 PO53-01 Saturday 340 2012


341 ENDO12L_SAT-300 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Resolution of Necrolytic Migratory Erythema with Octreotide in a Patient with Glucagonoma Adriana Martins Fernandes, Melina Silva Pinto, Daniel Soares Freire, Maria Adelaide Albergaria Pereira Hospital das Cl[iacute]nicas da Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Glucagonoma is a rare neuroendocrine tumor that arises from pancreatic [alpha] cells. In roughly 70% of cases, it is associated with a particular dermatosis called necrolytic migratory erythema. Other manifestations include diabetes mellitus, anemia, weight loss, diarrhea, deep vein thrombosis and neuropsychiatric disorders.[br]A 52-year old Caucasian man was referred to the dermatology department because of an extensive, itching cutaneous eruption that had been present for 2 years. The dermatosis initiated on lower extremities and progressed to involve trunk, arms, head and neck. Treatment with topical fluorinated corticosteroids, topical antibiotics or systemic steroids caused no improvement on skin lesions. In the meantime, diabetes mellitus was diagnosed and treatment with glyburide was initiated. In addition, the patient complained of weight loss (15 kg). A skin biopsy was consistent with necrolytic migratory erythema and the patient was transferred to the endocrinology department.[br]Laboratory analysis reviewed the following: hemoglobin, 11.3 g/dL; FPG, 198 mg/dL; HbA[sub]1C[/sub], 7.2%; albumin, 2.7 g/dL; serum glucagon, 1909 pg/mL (Ref.: [le]134 pg/mL); chromogranin A, 62 ng/mL (Ref.: 1.9-15 ng/mL); zinc, 27.5 [mu]g/dL (Ref.: 50-150 [mu]g/dL).[br]NMR imaging demonstrated a 6.7 cm nodular mass in the tail of the pancreas, which was hyperintense on T2 and exhibited enhancement after contrast injection. There was no evidence of metastases on the liver or elsewhere.[br]Treatment with octreotide (Sandostatin[reg], Novartis), 0.1 mg TID was initiated for three weeks, with substantial improvement of cutaneous lesions.[br]The patient underwent distal pancreatectomy with splenectomy. Histological examination revealed a neuroendocrine carcinoma with positive staining for glucagon, synaptophysine and chromogranin A. One week after surgery, the cutaneous lesions completely remitted.[br]The exact cause of the skin symptoms of necrolytic migratory erythema in the glucagonoma syndrome is not completed understood. The elevated glucagon levels stimulates carbohydrate metabolism, leading to hypovitaminosis B. Also, the catabolic state produced by the hyperglucagonemia leads to hypoamminoacidemia and deficiency in essential fatty acids. Zinc deficiency is another possible contributing factor. Normalization of glucagon levels, through tumor excision or pharmacological treatment with somatostatin analogues, results is resolution of the skin disorder, as seen in our patient.[br][br]Nothing to Disclose: AMF, MSP, DSF, MAAP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1306 59 402 SAT-300 PO53-01 Saturday 341 2012


342 ENDO12L_SAT-301 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Malignant Insulinoma in an Adolescent Girl Sookyoung Jeong, Ben Kang, Sohyun Park, Yongse Kwon, Soonki Kim, Yoonmee Choi, Joonmee Kim, Jieun Lee INHA University Hospital, Incheon, Republic of Korea; INHA University Hospital, Incheon, Republic of Korea; INHA University Hospital, Incheon, Republic of Korea An insulinoma is a rare pancreatic endocrine tumor that is typically sporadic, solitary, and less than 2 cm in diameter. Most insulinomas are benign with less than 10% demonstrating malignant behavior, the vast majority of which occur in adults. We report a case of malignant insulinoma in a 11-year-old girl. A girl presented with a 5-month history of recurrent episodes of seizures, especially in the morning. She had previously undergone Brain MRI and EEG studies, which revealed to be normal 2 months ago. On physical examination, her height was 161.3 cm ([gt]97 percentile), and her weight was 47.6 kg (75th). Her consciousness was alert and vital signs were normal. Physical exams of the orgens were also normal including no hepatosplenomegaly. Neurological exams were also normal. Laboratory test resuls during fasting showed marked hypoglycemia with a serum glucose level of 21 mg/dl coupled with an elevated serum insulin level of 50.1 uU/ml and elevated C-peptide level of 1.81 ng/ml. Urine test were negative for ketone. Glucagon stimulation test results revealed an increase of insulin level to 85.8 uU/mo after the infusion of glucagon. On radiologic imagin studies, 3[times] 2.4 cm size, solid mass with lobulated margin in the pancreatic tail was shown on MR images of the pancreas. The patient underwent laparoscopic distal pancreatectomy on the 12th hospital day. Fasting serum glucose concentrations were maintained at levels of 130-170 mg/dl after surgery, showing elevation compared to levels before surgery of 81-88 mg/dl only maintained by intravenous infusion of 10% dextrose fluid. The patient was discharged 20 days after surgery with euglycemia sate. Two month later follow up of abdominal MRI revealed the existence of multiple small metastatic nodular lesions in the liver.[br]Malignat inslinoma is extremely rare in children with only a few cases reported in literature. Managing this rare entity in children is challenging in terms of achieving the correct diagnosis, selecting therapy.[br][br]Nothing to Disclose: SJ, BK, SP, YK, SK, YC, JK, JL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1374 59 403 SAT-301 PO53-01 Saturday 342 2012


343 ENDO12L_SAT-302 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Low and Behold: Refractory Hypoglycemia in a Patient with Malignant Insulinoma Rommel R Geronimo, Markus D Agito, Daniela Ciltea, John M Moorman Akron General Medical Center, Akron, OH; Akron General Medical Center, Akron, OH; Northeast Ohio Medical University, Rootstown, OH Introduction: Insulinoma is a rare neuroendocrine tumor with a reported incidence of 4 cases per million a year(1). Less than 10 percent of insulinomas are malignant(2). Malignant insulinoma may cause severe hypoglycemia refractory to medical treatment.[br]Case Report: A 64-year old Caucasian male initially presented with elevated liver enzymes on routine laboratory testing. CT scan of the abdomen revealed soft tissue mass at the pancreatic head with hypodense lesions in the liver. Core biopsy of the liver lesions was compatible with low-grade neuroendocrine tumor. The patient was scheduled for Whipple[apos]s procedure, but during surgery, his tumor was deemed unresectable. Instead, he was treated with concomitant chemoradiotherapy. On repeat CT scan, progression of liver metastases was noted. The patient underwent hepatic artery embolization, which was complicated by formation of a hepatic abscess that required surgical drainage.[br]Two years after the initial presentation, the patient developed episodes of severe hypoglycemia. His work-up showed elevated insulin at 36mU/mL ([lt]17mU/mL), pro-insulin at 449.2pmol/L ([le] 18.8pmol/L) and C-peptide at 6.35mg/mL (0.80[ndash]3.10ng/mL). The patient was diagnosed with malignant insulinoma and received dextrose infusion, subcutaneous octreotide, oral diltiazem and diazoxide. Despite treatment, he continued to have severe hypoglycemic episodes requiring recurrent hospital admissions and increasing doses of diazoxide and octreotide. Oral prednisone was added with transient improvement in his blood glucose levels. Medical treatment was complicated by thrombocytopenia attributed to diazoxide. The patient[apos]s platelet count improved after diazoxide was discontinued. On his last admission the patient was discharged on an octreotide infusion pump and D20 intravenous fluid. He is scheduled for radioembolization of the hepatic lesions with Yttrium-90.[br]Discussion: This is a case of a 64-year old man who presented with pancreatic cancer, which turned out to be malignant insulinoma. This case illustrates how pancreatic neuroendocrine tumors, such as insulinoma, may initially present as nonfunctioning tumors but later result in hormone overproduction and difficult-to-treat hypoglycemia. Patients with unresectable insulinoma can be challenging to manage as they are usually refractory to medical treatment and require a multi-disciplinary approach.[br][br](1) Service FJ et al., Mayo Clinic Proceedings 1991; 66(7):711. (2) Hirshberg B et al., Cancer 2005; 104(2):264.[br][br]Nothing to Disclose: RRG, MDA, DC, JMM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1865 59 404 SAT-302 PO53-01 Saturday 343 2012


344 ENDO12L_SAT-303 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Insulinoma in a Patient with [ldquo]Reactive[rdquo] Hypoglycemia Aswathiah Srinath, Jack Martin, Alberti Isla, Abbi Lulsegged Kings College Hospital, London, UK; East and North Hertfordshire NHS Trust, Hertford, UK; North West London Hospitals NHS Trust, Harrow, UK; South London Healthcare NHS Trust, Orpington, UK 43year old healthy and active male presented with episodes of sweating, tiredness, difficulty concentrating [amp] recalling information lasting 10 to 15 minutes particularly between meals. He had difficulty to recall these events but never had any other neurological symptoms. Reflux symptoms was the only past medical history to note.[br]Examination: no features of endocrinopathy. BP 140/90, pulse 70 with waist circumference of 80cms. Systemic examination unremarkable.[br]He underwent prolonged glucose tolerance test which revealed blood glucose level of 1.4mmol/L ([lt]36mg/dl) at 5 hours at that time he became clammy found it difficulty to concentrate. Synacten test showed adequate steroid response of 896nmol/L at 60 minutes [amp] stool examination for H pylori was negative. At this stage a diagnosis of reactive hypoglycaemia was made. His symptoms improved on a low glycaemic diet.[br]Subsequently he had two similar episodes while he was on holiday with glucometer readings less than 2mmol/L ([lt]36mg/dl). There was no change in his diet at that time. Urgent admission for 72 hour fast revealed a glucose level of 1.7mmol/L ([lt]36mg/dl) associated with symptoms of hypoglycaemia and elevated levels of Insulin at 19.6U/L, C-Peptide 3.88mcg/L and ProInsulin 11pmol/L([lt]11pmmol/l). No further episodes were noted on Diazoxide 200mg BD.[br]Biochemical screening for MEN I was negative. Urinary [amp] plasma metanephrines were within normal range. Urine for sulphonylurea was negative.[br]CT showed a 10mm solid arterialised lesion in head of pancreas and there was a correlating lesion demonstrated on MRI. Endoscopic USS confirmed 16mm rounded solid lesion in head of pancreas. A diagnosis of Insulinoma was made and referred for endoscopic enucleation.[br]Conclusion: (1) Insulinoma can present with reactive hypoglycaemia and insulinomas should be considered if hypos persist despite change in diet or patient does not have risk factors or obvious causes for reactive hypoglycaemia. (2) Biochemistry during 72 hour fast may be mildly abnormal as in this case where insulin was only mildly elevated. This does not rule out insulinomas.[br][br]Postgraduate Medical Journal (October 1979)5,735-738. N Engl J Med.1989; 321(21): 1421.[br][br]Nothing to Disclose: AS, JM, AI, AL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1228 59 405 SAT-303 PO53-01 Saturday 344 2012


345 ENDO12L_SAT-304 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Cushing Syndrome Associated with pNET with Liver Metastasis: A Rare Presentation Enrica Salomone, Giovanna Muscogiuri, Clelia Cipolla, Caterina Policola, Silvia Della Casa, Chiara Maria Assunta Cefalo, Teresa Schiro, Anna Capozzi, Andrea Giaccari Catholic University, Rome, Italy Background.[br]NETs constitute a heterogeneous group of neoplasms that originate from endocrine glands, as well as endocrine islets within glandular tissue and cells dispersed between exocrine cells, such as endocrine cells of the digestive and respiratory tracts. NETs may present with a wide variety of functional or nonfunctional endocrine syndromes. Imaging tecniques include endoscopic US, CT, MRI and particularly, scintigraphy with somatostatin analogs and Gallium-68-DOTA-NOC PET/CT, wich appear to be a highly sensitive and specific tools for detecting of gastroenteropancreatic NET compared to conventional imaging such as CT and MRI.[br]Ectopic ACTH production by the pancreatic neuroendocrine tumor (p-NET) is relatively rare: in fact in literature case reports of Cushing[apos]s syndrome caused by ectopic ACTH secreting by pancreatic NET along with hepatic metastases with positive gallium-68-DOTA-NOC PET were not reported.[br]We report the case of a 43-year-old woman, who in September 2011, because of hirsutism, amenorrhea, muscle weakness, edema of bilateral lower extremities and face, underwent CT and US of abdomen, that demonstrated the presence of a 6-cm mass of the pancreas, associated with hepatic nodules.[br]On admission, she showed Cushingoid appearance; laboratory examinations revealed significant increase in plasma ACTH (260 pg/ml; normal range 10-55), cortisol (253 ng/ml; normal range 60-220) UFC: 1815.2 [mu]G/24 h (normal range [lt]70.0); 1-mg overnight DST: 281 ng/ml; 8 mg DST: 236 ng/ml and hypokalemia (2.8 mEq/l).[br]Were detected elevated levels of serum gastrin: 1976.0 pg/ml (normal range 30.0-115.0) and chromogranin A: 2031.7 ng/ml (normal range 19.4-98.1).[br]Histological examinations by needle biopsy of liver and pancreatic tumors indicated metastatic p-NET ACTH-producing; ACTH, synaptophysin and chromogranin A were identified by immunohistochemistry, and a Ki67 proliferation index was 5 %, NET, G2, (WHO 2010).[br]For a functional study of NET, the patient underwent a gallium-68-DOTA-NOC PET/CT which showed an increased uptake of the tracer receptor at the level of pancreas and hepatic nodules.[br]A case of p-NET associated with Cushing[apos]s syndrome is rare and in literature the detection of ectopic ACTH Cushing to pancreatic origin using PET with gallium-peptides was not reported.[br]The patient underwent steroid-blocking ketoconazole therapy, somatostatin analogue (lanreotide 120 mg), and she is waiting to perform FDG/PET/CT.[br][br]Nothing to Disclose: ES, GM, CC, CP, SDC, CMAC, TS, AC, AG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1474 59 406 SAT-304 PO53-01 Saturday 345 2012


346 ENDO12L_SAT-305 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Cyclical Cushing Syndrome (CCS): Clinical Inertia Despite Clear Diagnosis [mdash] Lessons for All Grace Sun, Allan Siperstein, Keith Peters, Rajesh Desai, Laurence Kennedy Cleveland Clinic Foundation, Cleveland, OH; University of Florida, Gainesville, FL; Medical Associates of Brevard, Melbourne, FL [bold]Background[/bold]: CCS is said to be rare, and cortisol periodicity is thought to hamper diagnosis. We present a case of CCS in which the diagnosis was clear, but definitive treatment was nevertheless delayed.[br][bold]Clinical Case[/bold]: For 5 years a 26-year old man had episodes of facial plethora and swelling, acne, peripheral edema, weight gain, hypertension and livid striae lasting 4-5 weeks with spontaneous regression. He felt well between episodes, which recurred every 5-6 months. During episodes, ACTH ranged from 145-1094 pg/mL, serum cortisol from 41.3-115.4 mcg/dL, and urinary free cortisol was as high as 9898 mcg/24h. Between episodes, ACTH and serum cortisol were 98-195 and 4.5-8.3, respectively. Pituitary MRI was normal. Treatment with ketoconazole did not affect his course. On referral to the Cleveland Clinic 5 years after symptom onset, he was nearing the end of an episode and was clearly Cushingoid on examination. Inferior petrosal sinus sampling suggested ectopic ACTH production. Chest CT showed a 2-cm anterior mediastinal mass with positive octreotide uptake. A thymic mass was removed; pathology showed a well-differentiated neuroendocrine carcinoma (atypical carcinoid), staining positively for ACTH, chromogranin and synaptophysin. He did not develop adrenal insufficiency and had recurrent, severe symptoms of hypercortisolism 5 months post-operatively. CT/PET scan revealed 2 metabolically active foci at the high left internal mammary chain and at the left lung hilum. Given the clinical situation, it was felt that bilateral adrenalectomy offered definitive treatment for the recurrent hypercortisolism; this was performed 6 years after the initial episode. Two months after adrenalectomy, he remains well on steroid replacement and is undergoing evaluation for further treatment directed at the thymic carcinoid metastases.[br][bold]Conclusions[/bold]: The episodic nature of the symptoms did not impair the diagnosis of CCS but may have contributed to clinical inertia. Prompt treatment of CCS is imperative since the cause may often be ectopic ACTH secretion from a potentially malignant tumor. In this case, bilateral adrenalectomy, rather than being a last resort, eliminated hypercortisolism as an ongoing clinical concern and shifted the focus to treatment of the primary tumor.[br][br]Nothing to Disclose: GS, AS, KP, RD, LK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 359 59 407 SAT-305 PO53-01 Saturday 346 2012


347 ENDO12L_SAT-306 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Surviving a Steroid Storm Noureddine Moumli, Gerlof Valk, M E Sanson-van Praag, Pierre MJ Zelissen UMC, Utrecht, Netherlands; Meander MC, Amersfoort, Netherlands [bold]Case:[/bold] A 60-year old previously healthy man was admitted with symptoms of weakness, weight gain and edema. Laboratory tests revealed a hypokalemia, hyperglycemia and metabolic alkalosis.[br]We tested and confirmed the diagnosis of ACTH-dependent Cushing syndrome: extremely elevated cortisol urinary excretion (42.000 nmol/24h, N 90-450), absent circadian rhythm of cortisol (4 umol/L, N 0.20-0.65), abnormal low-dose dexamethasone suppression test (2.74 umol/L, N [lt]0.05) and high ACTH levels (160-600 ng/L, N 10-70). MRI of the pituitary showed no abnormalities. High-dose dexamethasone-suppression and CRH-stimulating tests suggested the presence of an ectopic source of ACTH-production.[br]We performed an FDG-PET/CT-scan, which revealed a metabolic active retroperitoneal tumour near the duodenum and bilateral adrenal masses. A biopsy via endo-echography showed tumorous cells of neuro-endocrine origin, with positive ACTH-staining.[br]As the majority of gastroenteropancreatic neuro-endocrine tumours (GEP-NETs) express high concentrations of somatostatin-receptors, octreotide scintigraphy was performed, showing only pathologic uptake in the earlier described tumour, and a new smaller tumour along the duodenum.[br]The patient[apos]s condition deteriorated. He developed severe pancytopenia, pneumonia and progressive weakness. We started 2 inhibitors of cortisolsynthesis (ketoconazole and metyrapone) and octreotide to lower the ACTH production and release from the tumour. Within 8 days the cortisol levels dropped dramatically.[br]Subsequently the 2 tumours and left adrenal gland were resected. Pathologic examination revealed low-grade tumours (6.5 and 4 cm); the adrenal gland was hyperplastic with no signs of metastasis.[br]In the months after discharge the patient fully recoverd. Laboratory tests were normal. Repeated thoracal/abdominal CT-scans and endo-echography showed no abnormalities up to 8 months postoperatively.[br][bold]Discussion:[/bold] GEP-NETs are rare (incidence 2-4 per 100.000 people/year), and only a few produce ACTH. From 1968 to 2011 limited case-series were published, with a total of 51 patients. A hallmark of these tumours is the rapid deterioration due to overwhelming cortisol secretion with detrimental multiorgan effects and high mortality. We emphasize the importance of accurate assessment of the location and extent of the tumour, which is crucial for management. Furthermore we advocate aggressive and timely initiated multitargeted treatment to block the excessive cortisol production.[br][br]Nothing to Disclose: NM, GV, MES-v-P, PMJZ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2294 59 408 SAT-306 PO53-01 Saturday 347 2012


348 ENDO12L_SAT-307 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) A Rare Case of Cushing Syndrome Nisha Kaimal, Shankar Dhandapani, Sheila Grecian, Piotr Krysiak, Nauman Chaudhry, Basil Issa University Hosiptal of South Manchester, Manchester, UK; University Hospital of South Manchester, Manchester, UK; University Hospital of South Manchester, Manchester, UK [bold]Introduction[/bold]: Ectopic ACTH causes 10-15% of Cushing[apos]s syndrome. Bronchial carcinoids are the commonest cause, but other rare causes include small cell lung cancer, pancreatic tumours, medullary thyroid carcinoma, pheochromocytoma and thymic carcinoids (TC). We describe a rare case of ACTH producing TC presenting with type 2 diabetes and Cushing[apos]s syndrome.[br][bold]Clinical case[/bold]: A 49-year-old lady presented with type 2 diabetes and uncontrolled hyperglycemia, hypertension and hypokalemia. Cushing[apos]s syndrome was suspected because of the presence of central adiposity, round plethoric face and proximal myopathy. A random serum cortisol was elevated at [gt]1750 nmol/L and ACTH was 683ng/L ([lt]46ng/L). 24 hour urinary free cortisol was grossly elevated at 75,194 nmol/24 hours ([lt] 165nmol) and high dose dexamethasone suppression test failed to suppress cortisol values below 1750 nmol/L ([lt]50nmol/L). Because of the highly elevated cortisol levels and the presence of hypokalemia, ectopic ACTH secretion was suspected and a CT scan of her chest, abdomen and pelvis revealed a 55x24mm soft tissue mass in the anterior mediastinum, suggestive of a thymic lesion, and bilateral adrenal hyperplasia. MR scan of the pituitary was normal and octreotide scan results were concordant with the thymic lesion on CT. She was commenced on metyrapone, but deteriorated rapidly with septicaemia, cavitatory lung lesions, brain abscesses and superadded lung fungal infection. Despite being a high surgical risk, it was recognised that her infections would be difficult to control without treating her hypercortisolism and she underwent thoracotomy and thymectomy. Histology confirmed a paragangliod variant of TC with positive staining for ACTH. Cortisol and ACTH levels were restored to normal after surgery.[br][bold]Discussion: [/bold]Neuroendocrine carcinomas of the thymus account for 2-4% of all anterior mediastinal tumours. TC associated with ectopic ACTH secretion is very rare. When associated with an endocrinopathy, TC behave more aggressively. Our patient presented with type 2 diabetes, hypertension and obesity; conditions seen commonly by all clinicians. This case highlights a) the importance of having a high index of suspicion of Cushing[apos]s syndrome, combined with endocrine testing and imaging modalities to reach a diagnosis b) that timely institution of definitive treatment (in this case surgical removal of the source of ACTH production) is crucial in the context of life-threatening complications of the disease.[br][br]Nothing to Disclose: NK, SD, SG, PK, NC, BI 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 744 59 409 SAT-307 PO53-01 Saturday 348 2012


349 ENDO12L_SAT-308 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Double ACTH Secretion (Pituitary Adenoma and Pulmonary Carcinoid) Versus a Histological Pitfall in Cushing Syndrome [mdash] Case Report Juraj Payer, Peter Vanuga, Jana Kollerova, Eva Nemethova, Peter Jackuliak, Frantisek Ondrias University Hospital and Medical Faculty of Comenius University, Bratislava, Slovakia (Slovak Republic); National Institute of Endocrinology and Diabetology, Lubochna, Slovakia (Slovak Republic); Alpha Medical sro, Bratislava, Slovakia (Slovak Republic) Introduction: Cushing[apos]s syndrome is commonly caused by ACTH-secreting pituitary adenoma, but 1-10% is of ectopic origin from a pulmonary carcinoid tumor. Very rare cases of parallel presence of both are described in literature.[br]Clinical case: A 31-years-old man presented with typical symptoms of Cushing[apos]s syndrome with elevated serum and urinary levels of cortisol and serum levels of ACTH. No supressibility of serum cortisol levels was observed in dexamethason suppression test and diurnal periodicity was also impaired. MRI revealed a pituitary microadenoma, sinus petrosus venous sampling proved the pituitary origin of ACTH oversecretion. After hypophysectomy, the ACTH levels slightly declined, but remained elevated (up to 226 pg/ml - normal range up to 50 pg/ml). Due to this, multiple resections of the residual tissue were performed immunohistochemical methods repeatedly confirmed a positivity of ACTH and PRL production in the removed tissue. Due to the progressively impairing clinical condition of the patient a bilateral adrenalectomy was preformed from vital indication. Complete hormone replacement therapy was initiated and the clinical status of the patient markedly improved. Nevertheless, serum levels of ACTH remained high. During differential diagnostics a chest X-ray and CT showed a nonspecific finding in the upper lobe of the right lung suspicious of postinflammatory changes. No progression or changes were observed on regular controls until now. After 7 years the mentioned finding enlarged and the diagnosis of a primary typical carcinoid of the lungs with metastases to lymphnodes was confirmed by histology and immunohistochemistry. After surgical resection of the tumor ACTH levels normalised. The patient is undergoing radiotherapy and an octreoscan is planned. A second-look reevaluation of histological material from surgical interventions is in progress.[br]Conclusion: We present an extremely rare case of a patient with Cushing[apos]s syndrome with persistent high-levels of ACTH after hypophysectomy for an adenoma. 7 years after the primary diagnosis a pulmonary carcinoid was surgically removed with a consecutive normalisation of ACTH levels. Despite of the clear diagnosis of an ACTH-producing pituitary tumor, a double malignancy or an incorrect interpretation of histology can be assumed. Despite the clear identification of the primary source, in clinical practice a differential diagnostic process should be considered when ACTH levels remain elevated.[br][br]Nothing to Disclose: JP, PV, JK, EN, PJ, FO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1268 59 410 SAT-308 PO53-01 Saturday 349 2012


350 ENDO12L_SAT-309 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Pituitary Carcinoma with Diabetes Insipidus, Panhypopituitarism, Elevated Ki-67 and Metastases to an Unusual Site Varun Mehta, Laura Knecht St Joseph[apos]s Hospital, Phoenix, AZ [bold]Introduction:[/bold][br]Pituitary carcinomas constitute 0.1-0.2% of pituitary tumors. Clinical presentation is similar to that of an adenoma with visual changes, headache and effects of hormone hypersecretion. Presence of other CNS lesions or distant metastases is suggestive of a carcinoma. Gonadotroph carcinoma is even rarer with only a few published cases. It is usually characterized by elevated FSH and LH levels. Our patient was an unusual case, presenting with diabetes insipidus and a CNS lesion prior to noting spread to both adrenals, an uncommon site for metastases.[br][bold]Case Report:[/bold][br]A 58 year old male presented with failure to thrive. He had recently been diagnosed with a pituitary mass and was undergoing treatment for adrenal insufficiency, hypothyroidism and diabetes insipidus. MRI revealed a pituitary lesion with mass effect upon the optic chiasm and a ring-enhancing mass in the right frontal lobe. Both masses were surgically resected. Biopsy of the pituitary mass revealed a non-small cell malignant neoplasm consistent with pituitary carcinoma (gonadotropin). It was strongly positive for alpha subunit and weakly positive for luteinizing hormone immunoreactivity. The front mass had similar immunohistochemical features as the pituitary mass, indicating a metastatic spread from the primary pituitary neoplasm. Patient was started on cabergoline and temodar and underwent radiation treatment with good response. CT scan at 6 months showed incidental bilateral adrenal masses suspicious for metastases. Biopsy revealed it to be a neuroendocrine neoplasm originating from the pituitary with a Ki-67 labeling index (LI) of 60%. Currently the patient continues to undergo outpatient treatment with cabergoline, temodar, DDAVP, thyroxine and steroids.[br][bold]Discussion:[/bold][br]Primary pituitary carcinomas are rare tumors with about 140 documented cases. They are usually corticotroph tumors or prolactinomas but can occur in any cellular subtype. The hallmark of a pituitary carcinoma is distant metastases. Diagnosis cannot be made without evidence of metastatic disease, either outside the CNS or as separate noncontiguous foci within the CNS. Estimation of the Ki-67 LI using MIB-1 antibody has been shown to correlate best with invasiveness and prognosis. Treatment modalities include surgery, radiation and chemotherapy. Hormone hypersecretion, depending on the type of tumor can be managed with dopamine analogs or somatostatin. Treatment is mainly palliative with a mean survival time of 2 years.[br][br](1) Kaltsas, et al. Diagnosis and Management of Pituitary Carcinomas. Journal of Endocrinology and Metabolism. May 2005. 90(5):3089. (2) Ragel, et al. Pituitary carcinoma: a review of the literature. Neurosurg Focus 16(4). Article 7, 2004.[br][br]Nothing to Disclose: VM, LK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 232 59 411 SAT-309 PO53-01 Saturday 350 2012


351 ENDO12L_SAT-310 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Successful Treatment of Pituitary Carcinoma after Surgery, Radiotherapy and Temozolomide Cecile N Chougnet, Cyril Garcia, Lyse Bordier, Olivier Dupuy, Stephane Gaillard, Gerald Raverot, Olivier Bauduceau, Eric Baudin, Herve Mayaudon Institut Gustave Roussy, Villejuif, France; HIA Begin, Saint Mande, France; CH Foch, Suresnes, France; CHU Lyon, Bron, France; HIA Val de Grace, Paris, France The malignant prolactinoma is uncommon and treatment is not well established.[br]aims: We report a case of successful outcome after surgery and sequential treatment with temozolomide, during and after pituitary radiotherapy.[br]methods: A 44 year old man presented bilateral cavernous sinus invasion by a pituitary macroprolactinoma (28x28x48mm). After 6 months use of cabergoline, prolactine level dropped and tumor size decreased. But 9 months later secondary resistance was observed, despite increasing dose of cabergoline. An incomplete trans-sphenoidal debulking was performed. Tissue analysis showed a positive immunostaining for prolactine (90%) and GH (10%), with high proliferative index (mitotic count 20/10 HPF, Ki67 30%) leading to the diagnosis of malignant prolactinoma. No metastasis was revealed. The patient underwent radiotherapy (60 Gy) centered on the pituitary with concomitant administration of temozolomide. Then, he received 6 sequential cycles of temozolomide (150 mg/m[sup2]/d, 5 days every 4wk) with good tolerance.[br]results: A clear response was noted: prolactine level dramatically dropped with nearly complete tumor shrinkage within both cavernous sinus.[br]conclusion: Few cases of pituitary cancers are published. Objective response to temozolomide is inconstantly reported, perhaps due to its too late use or genetic polymorphisms. We propose to first use temozolomide during radiotherapy as this drug has radiosensitizing properties, then its classical administration.[br][br]Nothing to Disclose: CNC, CG, LB, OD, SG, GR, OB, EB, HM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2359 59 412 SAT-310 PO53-01 Saturday 351 2012


352 ENDO12L_SAT-311 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Unexpected Infertility in a Man with Acromegaly Leading to a Rare Diagnosis Jillene M Brathwaite, Ibrahim F Sherrif, Jeanne O[apos]Brien, Ismat Shafiq, Glynis Scott, Gabrielle A Yeaney, G Edward Vates, Laura M Calvi University of Rochester Medical Center, Rochester, NY; University of Rochester Medical Center, Rochester, NY; University of Rochester Medical Center, Rochester, NY; University of Rochester Medical Center, Rochester, NY; University of Rochester Medical Center, Rochester, NY [bold]Background[/bold]: Infertility is rarely reported in acromegaly. When seen, it is usually in conjunction with hyperprolactinemia, with hyperprolactinemia being the underlying cause for infertility, which is typically easily reversible [1]. In this case we highlight the importance of obtaining gonadotropins in patients with acromegaly who present with infertility.[br][bold]Clinical case[/bold]: We describe a 44 year old man who presented with a previous diagnosis of acromegaly, hyperprolactinemia and infertility. He was seen by a Urologist in his early teens and then an Endocrinologist 3-4 years prior to presenting to our center. His acromegaly was initially treated medically but patient discontinued treatment on his own after about 6 months as he did not really understand why he needed to take the medication. We diagnosed hypergonadotropic hypogonadism, which was not consistent with his acromegaly or hyperprolactinemia and thus prompted the search for an additional diagnosis. On exam, he was obviously acromegalic, displaying prognathism, frontal bossing and prominent zygomatic arches. He was also noted to have spotty pigmentation on the lips, buccal mucosa, face and trunk. On further questioning, he admitted to a prior diagnosis of hypogonadism but never pursued further work-up or treatment. Patient stated that he was never given a reason for his hypogonadism. He also admitted to having testicular [ldquo]lumps[rdquo]. He initially noted these around age 13 and had a testicular biopsy at that point. Based on the biopsy he was told that he had testicular calcifications but, as far as he could remember, he was not given a reason for them. In addition, he was not told if it might affect his future fertility. Pituitary MRI imaging showed a pituitary adenoma. Resection of that adenoma confirmed it to be a growth hormone and prolactin-secreting tumor. Histologic examination confirmed cutaneous myxomas, and blue nevi. Testicular and thyroid ultrasounds revealed testicular calcifications and multiple hypoechoic thyroid nodules respectively. Therefore the diagnosis of Carney complex was made.[br][bold]Conclusion[/bold]: Infertility is rarely reported in patients with acromegaly. When infertility is present, it should prompt a clinical investigation starting with checking gonadotropins. Carney complex is a very important diagnosis to make as it may predispose to an increased risk of various neoplasia or life threatening cardiac myxomas.[br][br](1) Singh P et al., J Hum Reprod Sci 2011; 4(2):102[ndash]103.[br][br]Nothing to Disclose: JMB, IFS, JO, IS, GS, GAY, GEV, LMC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1232 59 413 SAT-311 PO53-01 Saturday 352 2012


353 ENDO12L_SAT-312 POSTER SESSION: Neoplasia of Endocrine Tissues Case Reports (1:30 PM-3:30 PM) Parathyroid Carcinoma Presenting by Incidental Finding and after Ten Years of Hemodialysis: Two Cases Report Chih-Yiu Tsai, Chuen Hseuh, Kun-Ju Lin, Miaw-Jene Liou, Tzu-Chieh Chao, Yu-Yao Huang, Jen-Der Lin Chang Gung Memorial Hospital, Chang Gung University, Taoyuan Hsien, Taiwan; Chang Gung Memorial Hospital, Chang Gung University, Taoyuan Hsien, Taiwan; Chang Gung Memorial Hospital, Chang Gung University, Taoyuan Hsien, Taiwan; Chang Gung Memorial Hospital, Chang Gung University, Taoyuan Hsien, Taiwan Background: Parathyroid cancer is a rare cause of hyperparathyroidism. Pre-operative diagnosis and final pathology are difficult. Here we describe two cases of parathyroid carcinoma with atypical clinical presentations.[br]Case 1: A 45 years old male patient was found hypercalcemia (12.5 mg/dL, 7.9-9.9) and osteoporosis (T score = -3.2) during health examination. After referral to our endocrinological clinic, physical examination was negative finding but laboratory examinations showed hypercalcemia with normal serum albumin (4.5 g/dL, 3.5-5.5), low-level normophosphatemia (2.4 mg/dL, 2.4-4.7), and elevated serum i-PTH (851 pg/mL, 10-57). We arranged parathyroid ultrasonography and dual phase Tc-99m MIBI parathyroid scan, both revealed a lesion in the mid-to-lower left thyroid bed. A 6.0x5.0 cm tumor adhesive to left recurrent laryngeal nerve and esophagus was then disclosed by surgical intervention. Final pathology was parathyroid carcinoma with an invasive growth to surrounding fibrovascular tissue. Serum calcium (9.5 mg/dL) and i-PTH (38.7 pg/mL) showed normalization one year later. No evidence of recurrence was noted during 11 years follow-up until now.[br]Case 2: A 53 years old male had history of end stage renal disease on hemodialysis since 2000. He was found a palpable neck mass during physical examination in 2005, which was proved as one lobulated lesion of left thyroid lobe by ultrasonography. After ten years hemodialysis and follow-up, three parathyroid glands enlargement with refractory hyperparathyroidism (i-PTH: 3,081 pg/mL, 7-53; serum calcium: 10.8 mg/dL; serum phosphate: 6.6 mg/dL) developed. Besides, ultrasonography with fine needle aspiration of left thyroid lesion showed follicular neoplasm. Therefore, surgical intervention was arranged in July, 2011. The left lower parathyroid gland, 2.5x1.5x1.0 cm in size, was found firmly adhesive to thyroid tissue and surrounding strap muscle. Final pathology was parathyroid carcinoma with suspicious focal invasion. Postoperative follow-up revealed rebounding i-PTH (1,634 pg/mL) with normal serum calcium (8.4 mg/dL) and serum phosphate (3.3 mg/dL) in 6 months later, but no evidence of recurrence was noted.[br]Conclusion: Parathyroid cancer could present with incidental finding in routing health examination or long term hemodialysis patients. After total removal of malignant tumors, patients may follow with good prognosis.[br][br]Nothing to Disclose: C-YT, CH, K-JL, M-JL, T-CC, Y-YH, J-DL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1059 59 414 SAT-312 PO53-01 Saturday 353 2012


354 ENDO12L_SAT-325 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Bone Quality as Measured by Trabecular Bone Score (TBS) in Patients with Adrenal Incidentalomas with and without Subclinical Hypercortisolism Cristina Eller-Vainicher, Valentina Morelli, Fabio M Ulivieri, Serena Palmieri, Elisa Cairoli, Volha V Zhukouskaya, Rosa Pino, Antonella Naccarato, Alfredo Scillitani, Paolo Beck-Peccoz, Iacopo Chiodini Fondazione IRCCS C[agrave] Granda - Ospedale Maggiore Policlinico, Milan, Italy; Fondazione IRCCS C[agrave] Granda - Ospedale Maggiore Policlinico, Milan, Italy; Ospedale [ldquo]Casa Sollievo della Sofferenza[rdquo], IRCCS, San Giovanni Rotondo, Italy [bold]Background[/bold]. Patients with adrenal incidentalomas (AI) and subclinical hypercortisolism (SH) are at risk of fracture independently of bone mineral density (BMD) and possibly due to reduced bone quality. The Trabecular Bone Score (TBS) is a new method of gray-level texture measurement that can be extracted from DXA images, which has been suggested to consent the routine clinical evaluation of bone microarchitecture in post-menopausal osteoporosis. Scarce data are available regarding the use of TBS in patients with glucocorticoid-induced osteoporosis and no study evaluated its use in endogenous hypercortisolism. Therefore, the aim of the present study is to investigate the role of TBS in SH.[br][bold]Subjects and Methods[/bold]: In 102 AI patients, SH was diagnosed in the presence of [ge]2 out of: urinary free cortisol [gt]70 [mu]g/24h (193.1 nmol/L), cortisol after 1[ndash]mg dexamethasone test (1mg-DST) [gt]3.0 [mu]g/dL, (82.8 nmol/L), ACTH [lt]10 pg/mL ([lt]2.2 pmol/L). By Dual X-ray Absorptiometry, BMD was measured at spine (LS) and femur (neck, FN and total, FT) and TBS was assessed in the same regions as for LS-BMD; BMD and TBS data were reported as Z-scores. The presence of morphometric vertebral deformity was assessed by thoraco-lumbar (T4-L4) radiograph.[br][bold]Results[/bold]. Age, BMI and gender were comparable among patients with (n=34) and without (n=68) SH. Patients with SH had lower LS-BMD (-0.31[plusmn]1.17), FT-BMD (-0.29[plusmn]0.91) and TBS (-3.18[plusmn]1.21) than patients without SH (0.31[plusmn]1.42, P=0.031; 0.19[plusmn]0.97, P=0.018; -1.71[plusmn]1.54, P[lt]0.0001, respectively). TBS was inversely correlated with 1mg-DST ([beta]= -0.26, t= -2.79, P=0.006) regardless of age, LS-BMD, BMI and gender.[br]Fractures were predicted by the presence of low TBS alone (OR=4.8, 95% CI=1.85-12.42, P=0.001) and the combination low TBS [italic]plus[/italic] low LS-BMD (OR=4.37, 95% CI=1.71-11.4, P=0.002) as well as by the absence of the combination normal TBS [italic]plus[/italic] normal LS-BMD (OR=4.39, 95% CI=1.58-12.18, P=0.004), but not by the presence of low LS-BMD alone, after adjustment for age, BMI and gender. Low TBS [italic]plus[/italic] low LS-BMD and normal TBS [italic]plus[/italic] normal LS-BMD showed good specificity for individuating fractured (79.1%) and not fractured (88.1%) patients, respectively. Finally, TBS predicted the occurrence of a new fracture in 40 AI patients followed up for 24 months (OR=11.2, 95%CI=1.85-1.71, P=0.012) after adjusting for LS-BMD, BMI and age.[br][bold]Conclusions[/bold]: In SH bone microarchitecture, as measured by TBS, is altered and TBS is useful in detecting AI patients at risk of fractures.[br][br]Nothing to Disclose: CE-V, VM, FMU, SP, EC, VVZ, RP, AN, AS, PB-P, IC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 490 60 415 SAT-325 PO05-01 Saturday 354 2012


355 ENDO12L_SAT-326 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Osteoporosis and Vertebral Fractures in Primary Aldosteronism Iacopo Chiodini, Antonio S Salcuni, Serena Palmieri, Vincenzo Carnevale, Valentina Morelli, Claudia Battista, Vito Guarnieri, Giuseppe Guglielmi, Gaetano Desina, Cristina Eller-Vainicher, Paolo Beck-Peccoz, Alfredo Scillitani Fondazione Policlinico, Mangiagalli e Regina Elena, IRCCS, University of Milan, Milan, Italy; [apos]Casa Sollievo della Sofferenza[apos], IRCCS, San Giovanni Rotondo, Italy; [apos]Casa Sollievo della Sofferenza[apos], IRCCS, San Giovanni Rotondo, Italy; [apos]Casa Sollievo della Sofferenza[apos], IRCCS, San Giovanni Rotondo, Italy; [apos]Casa Sollievo della Sofferenza[apos], IRCCS, San Giovanni Rotondo, Italy; [apos]Casa Sollievo della Sofferenza[apos], IRCCS, San Giovanni Rotondo, Italy [italic]Background[/italic]: In rats with aldosteronism, an increase in urinary calcium excretion with a reduction of bone mineral density (BMD) and cortical bone strength has been reported. Our study was aimed to evaluate bone involvement in patients with primary aldosteronism (PA).[br][italic]Methods[/italic]: One hundred eighty-eight consecutive subjects with adrenal incidentaloma, observed between November 2009 and October 2011, were screened for PA with aldosterone-to-renin ratio (ARR). After confirmatory tests, eleven patients were diagnosed as PA while fifteen patients were not (nPA). A serum/urinary biochemical profile, PTH, BMD measured at lumbar spine (LS) and femoral neck (FN and TN) by dual X-ray absorptiometry and conventional spinal radiographs in lateral (T4-L4) and anteroposterior projection were obtained in all subjects.[br][italic]Results[/italic]: PA patients had a significantly higher 24h urinary calcium (M[plusmn]SD) (251[plusmn]74 vs 171[plusmn]47 mg/die; p[lt] 0.01), PTH [Median (range)] 90.0 (53-134) vs 48.6 (23-99) pg/mL; p[lt] 0.01) than nPA patients. BMD (M[plusmn]SD) expressed as Z-value at LS (-1.18[plusmn]0.99 vs 0.22[plusmn]1.12), FN (-0.85[plusmn]0.73 vs 0.01[plusmn]0.82) and TN (-0.49[plusmn]0.61 vs 0.39[plusmn]0.93) was lower in PA than in nPA, p= 0.003, p= 0.011, p= 0.012, respectively. The prevalence of osteoporosis was higher in PA than in nPA (8/11, 72.7% vs 3/15, 20.0%; Fisher exact test: p= 0.015). Vertebral fractures tended to be more prevalent in PA than in nPA (5/11, 45.5% vs 2/15, 13.3%; Fisher exact test: p= 0.095). Bivariate correlations showed that in the all sample there was a significant association between: aldosterone and 24h urinary calcium (r= 0.43, p= 0.029); aldosterone and PTH (r= 0.42, p = 0.035); aldosterone and BMD measured at FN (r= -0.58, p [lt] 0.01) or LS (r= -0.66, p [lt] 0.01) or TN (r= -0.66, p [lt] 0.01); 24h urinary calcium and PTH (r= 0.57, p [lt] 0.01). Logistic regression analysis showed that osteoporosis and morphometric vertebral fractures were associated with PA (OR= 15[middot]4, C.I.= 1.83-130, p= 0.012; and OR= 30.4, C.I.= 1.07-862, p= 0.045, respectively) regardless for age, BMI and LS-BMD.[br][italic]Conclusions[/italic][bold]: [/bold]PA is associated with osteoporosis, vertebral fractures and increased urinary calcium excretion.[br][br]Nothing to Disclose: IC, ASS, SP, VC, VM, CB, VG, GG, GD, CE-V, PB-P, AS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1101 60 416 SAT-326 PO05-01 Saturday 355 2012


356 ENDO12L_SAT-327 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Occult Secondary Osteoporosis: Extensive Clinical Experience Cristina Eller-Vainicher, Elisa Cairoli, Volha V Zhukouskaya, Valentina Morelli, Serena Palmieri, Paolo Beck-Peccoz, Iacopo Chiodini Fondazione IRCCS C[agrave] Granda - Ospedale Maggiore Policlinico, University of Milan, Milan, Italy [bold]Background: [/bold]The prevalence of secondary osteoporosis (OP2) is debated. Only some causes may be identified by medical history and/or physical examination. To identify hidden causes of OP2 is important, since a missed diagnosis may lead to treatment failure. The aim of the study was to evaluate the prevalence and causes of hidden OP2 in a large sample of patients evaluated at the same center.[br][bold]Subjects and Methods. [/bold]Among 993 new consecutive patients evaluated for reduced bone mineral density and/or fragility fractures, 419 patients were excluded because of already known causes of OP2. The remaining 574 patients (539 women, 35 men, age 65.4[plusmn]10.1 years) underwent laboratory investigations (serum and 24h urinary calcium, 25-hydroxyvitamin D, creatinine, PTH, TSH, protein electrophoresis, IgA, anti-transglutaminase and endomisium antibodies, serum electrophoresis, C-reactive protein, cortisol after 1-mg dexamethasone test and serum testosterone in men) before and after vitamin D supplementation. All patients were classified as osteoporotic (T-score [le] -2.5) or osteopenic (T-score [le] -1.0), considering the worst T-score between lumbar spine and hip, on the basis of the DXA results. The vertebral deformities were assessed by radiograph, and each vertebra was defined as intact (grade 0) or grade 1 (20[ndash]25%), 2 (25[ndash]40%), or 3 ([gt]40%) deformity. The Spinal Deformity Index (SDI) was calculated by summing the grade of deformity for each vertebra.[br][bold]Results: [/bold]Hidden OP2 were diagnosed in 43.7% (251/574) of patients with apparently primary osteoporosis, due to: idiopathic hypercalciuria in 34.8% (n=201), primary hyperparathyroidism in 4.5% (n=26), hyperthyroidism in 1.2% (n=7), male hypogonadism in 5.7% (n=2/35), celiac disease in 1.4% (n=8), subclinical hypercortisolism 1.4% (n=8). Moreover, 70.6% of patients presented serum 25-hydroxyvitaminD [lt]30 ng/dl, and 11.2% [le]10 ng/ml. The 53.4% of patients presented [ge]1 vertebral deformity and 15.0% a SDI[ge]5. Patients with hidden OP2, were younger (63.8[plusmn]10.3 vs 66.7[plusmn]9.8 years, p=0.001) and had higher prevalence of osteoporosis (82.9% vs 72.4%, p=0.004) than patients with idiopathic osteoporosis, while no difference was found in gender, SDI, prevalence of vertebral deformities and hypovitaminosis D.[br][bold]Conclusions: [/bold]Almost 40% of patients with apparently idiopathic osteoporosis are affected with hidden OP2, with no difference between males and females. In about 30% of patients, osteoporosis is due to idiopathic hypercalciuria.[br][br]Nothing to Disclose: CE-V, EC, VVZ, VM, SP, PB-P, IC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1451 60 417 SAT-327 PO05-01 Saturday 356 2012


357 ENDO12L_SAT-328 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Nonalcoholic Fatty Liver Disease as a Risk Factor for Low Bone Mass in Postmenopausal Women Seong-Su Moon, Young-Sil Lee Dongguk University School of Medicine, Gyeongju, Korea Purpose: Osteoporosis is a disease associated with insulin resistant states such as central obesity, diabetes and metabolic syndrome. Nonalcoholic fatty liver disease (NAFLD) is also increased in such conditions. However, little is known about whether osteoporosis and nonalcoholic fatty liver disease are etiologically related to each other or not. We examined whether bone mineral density (BMD) is associated with NAFLD in pre- and postmenopausal women.[br]Methods: Four hundred eighty one female subjects (216 premenopausal, 265 postmenopausal) were enrolled. Lumbar BMD was measured using dual-energy X-ray absorptiometry (DXA). Liver ultrasonography was done to check the severity of fatty liver. We excluded subjects with a secondary cause of liver disease. Blood pressure, lipid profile, fasting plasma glucose, alanine aminotransferase (ALT), aspartate aminotransferase, and body mass index were measured in every subjects.[br]Results: Mean lumbar BMD was lower in subjects with NAFLD than those without NAFLD in postmenopausal women (0.98[plusmn]0.01 vs. 1.01[plusmn]0.02 g/cm2, P=0.046). Multiple correlation analysis revealed a significant association between mean lumbar BMD and NAFLD in postmenopausal subjects after adjusting for age, body mass index, ALT, smoking status, alcohol consumption ([beta] coefficient -0.066, 95% CI -0.105[sim] -0.027, P=0.001). Even after adjusting the presence of metabolic syndrome, the significance was maintained ([beta] coefficient -0.043, 95% CI -0.082 [sim] -0.004, P=0.031).[br]Conclusion: Lumbar BMD is related with NAFLD in postmenopausal females. We suggest that postmenopausal women with NAFLD may have a higher risk of osteoporosis than those without.[br][br]Nothing to Disclose: S-SM, Y-SL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1361 60 418 SAT-328 PO05-01 Saturday 357 2012


358 ENDO12L_SAT-329 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Efficacy and Safety of the Anabolic Therapies in Severe Osteoporosis: Experience of a Team of Endocrinologists and Spine Surgeons Marilda Mormando, Alessandra Fusco, Enrico Pola, Serena Piacentini, Debora Colangelo, Luigi Nasto, Mariella Celico, Antonio Bianchi, Alfredo Pontecorvi, Laura De Marinis Catholic University, Rome, Italy; Department of Orthopaedics and Traumatology, Catholic University, Rome, Italy [bold]Introduction[/bold]: Anabolic therapies represent a major advance in the management of osteoporosis, and they may provide significant benefit to those patients with severe osteoporosis in whom antiresorptive therapy has proven insufficient. Parathyroid hormone (PTH) and human recombinant parathyroid hormone peptide 1-34 (Teriparatide) demonstrated an increase in bone mineral density and a significant reduction in vertebral fractures in patients with osteoporosis when given for 18-24 months. The intermittent administration of parathyroid hormone (PTH) and teriparatide stimulates osteoblastic function, improves architecture at both the trabecular and cortical bone and has an additional analgesic effect.[br][bold]Materials and methods[/bold]: we retrospectively analyzed the safety, efficacy and adherence to therapy with anabolic agents given for 18 months in 53 patients (M/F: 7/46) with severe primary osteoporosis resistant to antiresorptive therapy. Patients were followed at an outpatient centre of osteoporosis disease by a team of spine surgeons and endocrinologists from 2007 to 2010. Before anabolic therapy 30 patients underwent percutaneus kyphoplasty and 23 were treated with brace. The anabolic agents were administered as a daily injection, and supplementation of vitamin D was given, when necessary. The change in BMD value was measured at the beginning of therapy and after 18 months through lumbar and femoral DEXA scan.[br][bold]Results[/bold]: A total of 45 patients completed the 18 months treatment with the anabolic agents. Then, six patients (11%) discontinued the treatment due to side effects occurring during the first 3 months of therapy (in detail, hypercalcemia occurred in 5 patients, and nausea in 1 patient). In 2 cases, the treatment was interrupted due to non adherence to therapy. All patients had vertebral fractures (VF) (mean number of VF 3.5[plusmn]1.5) and suffered from chronic and/or acute lumbar pain. After the beginning of the anabolic therapy, 47% of patients showed a significant improvement of lumbar pain. At the end of the 18 months of treatment, there was a substantial increase in BMD at both lumbar spine (+9%) and femoral neck (+5%).[br][bold]Conclusions[/bold]: This retrospective study confirms that the anabolic agents are safe and effective in the treatment of severe osteoporosis. Adherence to therapy is very high, despite the daily administration. Of note, this therapy can reduce the lumbar pain in a significant percentage of patients with multiple VF.[br][br]Nothing to Disclose: MM, AF, EP, SP, DC, LN, MC, AB, AP, LDM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1211 60 419 SAT-329 PO05-01 Saturday 358 2012


359 ENDO12L_SAT-330 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Effects of Teriparatide Treatment on Anabolic Hormones and Quality of Life in Patients with Osteoporosis with Vertebral Fractures Ece Harman, Korhan Baris Bayram, Gokcen Unal Kocabas, Ilgin Yildirim Simsir, Hasan Kamil Sucu Ataturk Training and Research Hospital, Izmir, Turkey; Ataturk Training and Research Hospital, Izmir, Turkey; Ataturk Training and Research Hospital, Izmir, Turkey; Ege University School of Medicine, Izmir, Turkey Objective: The aim of this cross-sectional study was to compare the effects of teriparatide therapy on anabolic hormones and bone turnover markers and quality of life in patients with osteoporotic fractures.[br][bold]Materials and methods:[/bold][br]Patients with severe osteoporosis defined as lumber vertebral total or femur total T score [le] -4.0 and at least 2 lumber vertebral compression fractures were included. Vertebral fracture was defined as loss of at least %25 height compared with adjacent vertebral height at lateral thoracal or lumber vertebral X-ray. 15 patients recently had teriparatide therapy and 15 patiens had no therapy for osteoporosis. Patients with thyroidal, parathyroidal, hepatic, renal disease, inflammatory joint disease, cancer, using drugs that effect bone metabolism and with traumatic vertebral fractures were excluded. Alcaline phosphatase, osteocalcin, GH, IGF-1, IGFBP and SHBG levels were determined. QUALEFFO was used for QOL evaluation.[br][bold]Results: [/bold]Age, height, weight, number of parturition and duration of menopause did not differ within groups. (p[gt]0,05) Nursing, immobilisation, smoking, coffee consumption and calcium intake also was not different (P[gt]0,05).[br]In the teriparatide group ALP and osteocalcin levels were significantly higher (p[lt]0,05)than no treatment group while GH, IGF-1, IGFBP and SHBG levels did not differ significantly. (P[gt]0,05) In teriparatide patients physical function (a,b,c,d,e), general health evaluation (f) and total QUALEFFO scores were significantly lower, while mental function (g) scores were significantly higher (p[lt]0,05). Physical function scores b and c were negatively correlated with IGF-1 (p[lt]0,05). Treatment duration in teriparatide group was negatively correlated with physical function score a,b,c,d ,and general health score (f) (p[lt]0,05).[br][bold]Conclusions:[/bold] In this study teriparatide therapy in osteoporotic patients with vertebral fracture was shown to siginificantly raise the levels of bone turnover markers and positively effect quality of life, and this situation was correlated with treatment duration.[br][br]Nothing to Disclose: EH, KBB, GUK, IYS, HKS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 456 60 420 SAT-330 PO05-01 Saturday 359 2012


360 ENDO12L_SAT-331 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) The Effect of Combined Raloxifene and Conjugated Estrogen on Bone Metabolism, Endometrial Safety, and Coagulation Profile in Postmenopausal Osteoporotic Women Hajime Oishi, Tetsu Yano, Osamu Hiraike-Wada, Mana Hirano, Yutaka Osuga, Shiro Kozuma, Yuji Taketani Faculty of Medicine, The University of Tokyo, Tokyo, Japan Context: It has recently been proposed that the combination of a selective estrogen receptor modulator (SERM) with estrogen may be an alternative to combination estrogen and progestin therapy (EPT) to prevent osteoporosis in at-risk postmenopausal women without the potential concerns associated with EPT.[br]Objective: The aim of the present study is to compare effects of 24 weeks[apos] treatment with either low-dose, alternate-day, estrogen-progestin or low-dose estrogen plus raloxifene (RLX) 60mg/day on bone mineral density, biochemical markers of bone turnover, and endometrial safety in postmenopausal osteopenic women.[br]Design: In this randomized controlled pilot study, postmenopausal women (46-62 yr of age) with osteopenia or osteoporosis, were treated with 1) conjugated estrogen 0.625mg plus medroxyprogesterone acetate 2.5mg on alternate days (CE/MPA; n=17) or 2) CEE 0.625mg on alternate days plus RLX 60mg/day (CE/RLX; n=15). Subjects were 1-10 years postmenopause, with uterus intact, and had a screening BMD T-score at lumbar below -1. The primary outcome was change in bone mineral density of lumbar spine. Other assessments include change in bone turnover markers, endometrial thickness, and D-dimer.[br]Results: In both groups, BMD increased significantly compared with baseline. Change in lumbar spine BMD increased significantly higher in CE/RLX group than CE/MPA group. Serum bone alkaline phosphatase and urine N-telopeptide significantly decreased with both groups. Endometrial thickness and level of D-dimer did not significantly change in both groups.[br]Conclusion: Combination CE (0.625mg, alternate-day) plus RLX (60mg per day) is an effective and safe treatment option for osteoporosis prevention. CE/RLX represents a novel alternative to traditional hormone therapies and further investigation will be needed.[br][br]Sources of Research Support: This work is supported by Grant-in-Aid for Scientific Research 23791815 from the Ministry of Education, Science, and Culture, Japan.[br][br]Nothing to Disclose: HO, TY, OH-W, MH, YO, SK, YT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1236 60 421 SAT-331 PO05-01 Saturday 360 2012


361 ENDO12L_SAT-332 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Effect of Bisphosphonate and Raloxifen on 2-Year Progression of Aortic Calcification Kwang Joon Kim, Sung-Kil Lim, Kyoung Min Kim, Yumie Rhee, Eun Young Lee Severance Hospital, Yonsei University of College of Medicine, Seoul, Korea; Yonsei University Health System, Seoul, Korea There exists a strong correlation between low bone mineral density and vascular calcification. The mechanism of vascular calcification has similarities to the process of osteogenesis. Thus, bisphophonate or hormonal replacement therapy has been considered to affect vascular calcification. We investigated the effect of anti-resorptive drug, risedronate and raloxifen, on vascular calcification. In postmenopausal women (n=38) with anti-resorptive therapy, quantitative computed tomography was performed for measuring bone mineral density. Aorta calcification score (ACS) were measured by the Agaston scoring method for 2 years. There was no difference of baseline characteristics of the two groups except c-terminal telopeptide level (Table 1). ACS increased with time. ACS increased more in risedronate group than in raloxifen group, but not significant (Table 2). ACS showed negatively correlation with bone mineral density. In multiple regression analysis, age only affected the change of ACS (Table 3). In conclusion, there was no difference in effect of the risedronate or raloxifen on ACS.[br][br]Nothing to Disclose: KJK, S-KL, KMK, YR, EYL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1583 60 422 SAT-332 PO05-01 Saturday 361 2012


362 ENDO12L_SAT-333 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Testosterone Replacement on Top of Bisphosphonate Therapy Has an Additional Benefit on Bone Mass, Libido and Sexual Activity in Male Cardiac Transplant Patients: A 5-Year Prospective Controlled Study Doris Wagner, Guenther Prenner, Harald Dobnig, Thomas Pieber, Barbara Obermayer-Pietsch, Andreas Tomaschitz, Astrid Fahrleitner-Pammer Surgery, Graz, Austria; Internal Medicine, Graz, Austria; Internal Medicine, Graz, Austria Hypogonadism is frequently encountered in cardiac transplant patients (HTX) and exerts negative effects not only on bone metabolism but also libido and quality of life. We investigated whether testosterone replacement therapy (TRT) on top of parenteral ibandronate (IBN) therapy in hypogonadal HTX recipients confers positive effects on bone mass, fracture incidence and quality of sex life compared to untreated hypogonadal and eugonadal HTX patients receiving solely quarterly IBN injections.[br]69 male patients with osteoporosis who were at least in the 2[sup]nd[/sup] year since surgery were screened, finally 52 patients entered the study. 31 (60%) were classified as hypogonadal, fourteen (45%) out of the hypogonadal patients additionally received testosterone replacement therapy on top of IBN. All patients received osteoprotective therapy with intravenous IBN (2mg every 3 months) as well as 1200mg calcium and 880IU vitamin D3.[br]At baseline 77% of the hypogonadal patients reported loss of libido (compared to 27% of eugonadal men, P=0.005) with an average of 7[plusmn]6 annual sexual activities (as compared to 15[plusmn]14 of eugonadal men, P=0.005). Hypogonadal men at baseline had also markedly lower Z-score values at the femoral neck (-1.54 vs. 0.15), and total hip (-1.34 vs. 0.01) (all P=0.0001) as well as a higher percentage of prevalent vertebral fractures (63.3%; vs. 13.6%; P=0.0003). After 1 and 5 years of treatment bone mineral density (BMD) had significantly increased in all patients, however hypogonadal patients with additional TRT had a significant higher BMD increase (neck 12.4 and 16.4%, total hip 9.2 and 12.4%, respectively, all P[lt]0.001) as compared to eugonadal patients (neck 2.9 and 3.4%, total hip 3.7 and 4.4%) and unreplaced hypogonadal patients (neck 2.3 and 3%, total hip 3.2 and 4.2%). Compared to baseline patients with TRT reported an increase in annual sexual activities after 1 (29[plusmn]8; p[lt]0.0001) and 5 years (25[plusmn]9; p[lt]0.0005). No changes in sexual behavior were reported by the other groups.[br]Hypogonadismus has a deleterious effect on bone health in transplant patients and is a complication among long term survivors. Intravenous IBN therapy increases BMD in HTX patients on continuing immunosuppressive treatment independently of gonadal status. Hypogonadal patients clearly benefit from additional TRT over at least 5 years with respect to changes in bone mass as well as quality of life. TRT and IBN are safe and well tolerated treatment options in this entity.[br][br]Nothing to Disclose: DW, GP, HD, TP, BO-P, AT, AF-P 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1563 60 423 SAT-333 PO05-01 Saturday 362 2012


363 ENDO12L_SAT-334 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Management of Osteoporosis in an Elderly Woman with Monoclonal Gammopathy Ambreen Qureshi, Hanna Mawad University of Kentucky, Lexington, KY; University of Kentucky, Lexington, KY [bold]Background[/bold]: Pharmacologic therapies for osteoporosis include anabolic agent teriparatide and antiresorptive drugs including bisphosphonates and denusomab. Choice of therapy should be made after assessing patient[apos]s fracture risk and comorbid conditions, and after determining whether the patient has low or high bone turnover as indicated by biochemical bone markers, and the results of a bone biopsy if it has been performed. [bold]Clinical Case[/bold]: An 87-year-old female patient presented for a second opinion regarding management of osteoporosis. Past medical history was significant for neuropathy, and osteoporosis diagnosed ten years ago. She had sustained multiple fractures despite treatment with various oral bisphosphonates for nine years. She was taking risedronate. Her last DEXA scan six months ago showed a T-score of -4.1 in L1 to L4,-3.2 in the left hip and -2.9 in the right hip.Risedronate was discontinued and bone markers were ordered:bone specific alkaline phosphatase,N-telopeptide collagen x-links and osteocalcin. These were not suppressed as expected with prolonged bisphosphonate use. Due to the patient[apos]s age, SPEP and UPEP were also ordered. UPEP was normal but SPEP demonstrated monoclonal gammopathy with elevated atypical IgM monoclonal protein, and marginally increased from one year ago. After extensive communication with the patient, her PCP and neurologist, it was determined that her neuropathy was secondary to monoclonal gammopathy and she had been treated with rituximab by a hematologist in the past. Bone biopsy was not performed as the patient could not be treated with teriparatide due to relative contraindication in premalignant conditions,even if the results had demonstrated high bone turnover requiring anabolic therapy. Denosumab is a monoclonal antibody against RANKLwhich has an additional role in the immune system;therefore its inhibition can have potential infectious and/or neoplastic complications. Since the patient had been treated with rituximab previously, denusomab could enhance her long-term risk of infections and malignancies, and it was decided to treat her with zoledronic acid instead of denusomab. [bold]Conclusion[/bold]: Each pharmacologic agent for osteoporosis has side effects. Until sufficient long-term safety data are available, clinicians should consider potential complications and make a choice after weighing the risks and benefits of each agent in each patient after careful assessment of his fracture risk and comorbid illnesses.[br][br]Nothing to Disclose: AQ, HM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2312 60 424 SAT-334 PO05-01 Saturday 363 2012


364 ENDO12L_SAT-335 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Nitroglycerin May Improve Bone Mass by Suppressing Sclerostin Levels Aarthi Arasu, Celeste J Hamilton, Steven R Cummings, Richard Eastell, Sophie A Jamal University of California, San Francisco, CA; University of Toronto, Toronto, Canada; California Pacific Medical Center Research Institute, San Francisco, CA; University of Sheffield, Sheffield, UK; University of Toronto, Toronto, Canada Context: Nitroglycerin, a nitric oxide (NO) donor, is a novel treatment for osteoporosis which stimulates bone formation and inhibits bone resorption. A recently published randomized control trial demonstrated that nitroglycerin ointment (15 mg/day) increases bone mineral density (BMD) in the lumbar spine (7%), total hip (6%), and femoral neck (7%) as well as indices of bone strength such as cortical thickness (14% at radius and 25% at tibia) in postmenopausal women after 2 years. However, the mechanism of this outcome is unknown. The effects of mechanical loading on the osteocyte to increase bone mass may be mediated by NO. Therefore, a potential pathway is that NO lowers the production of sclerostin, a protein secreted by osteocytes which inhibits bone formation.[br]Objective: To determine if nitroglycerin treatment of postmenopausal women decreases circulating sclerostin levels.[br]Design, Setting, Participants: A single-center, double-blind, placebo-controlled randomized trial was completed in ambulatory postmenopausal women with BMD T scores above -2.0 at the spine and hip. The study was conducted in Toronto, Ontario, Canada. Circulating sclerostin levels were measured by the Biomedica ELISA assay in a random sample of 25 women in the treatment group and 25 women in the placebo group at baseline and year 2.[br]Main Outcome Measure: Change in serum sclerostin levels[br]Results: The baseline characteristics of the random subsamples of women did not differ significantly such as age (60.5 v. 61.0 years), BMD (spine 1.05 v. 1.06 g/cm[sup]2[/sup]; total hip 0.91 g/cm[sup]2 [/sup]in both), weight (69.0 v. 71.3 kg), and walking for exercise (84% in both) in the placebo and treatment groups, respectively. Circulating sclerostin levels decreased significantly in the nitroglycerin treated participants by 21% (baseline, 32.8 pmol/l; year 2, 25.6 pmol/l; p = 0.01) but did not change in the placebo group (-6%; baseline, 26.7 pmol/l; year 2 24.6, pmol/l; p = 0.30). The percent decrease in circulating sclerostin levels after 2 years differed significantly between the treatment and placebo groups (p [lt]0.001).[br]Conclusion: Our results suggest that nitroglycerin treatment substantially increases bone mass by decreasing sclerostin production in postmenopausal women.[br][br]Sources of Research Support: Canadian Institute of Health Research; Physicians[apos] Services Incorporated.[br][br]Nothing to Disclose: AA, CJH, SRC, RE, SAJ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 762 60 425 SAT-335 PO05-01 Saturday 364 2012


365 ENDO12L_SAT-336 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Unfavorable Impacts of Aromatase Inhibitors on Cortical Geometry in the Femoral Neck in Postmenopausal Women with Breast Cancer Su Jin Lee, Kyoung Min Kim, Ji-Hae Shin, Sung-Kil Lim, Byeong Woo Park, Yumie Rhee Yonsei University, College of Medicine, Seoul, Republic of Korea; Yonsei University, College of Medicine, Seoul, Republic of Korea; Yonsei University, College of Medicine, Seoul, Republic of Korea The negative impact of aromatase inhibitors (AIs) used as adjuvant hormonal therapy on bone mass determined by dual energy X-ray absorptiometry and increased risk of fragility fractures in postmenopausal women with breast cancer are obvious. As fracture risks highly depend on the bone structure as well as bone mass, we have adapted quantitative computed tomography (QCT) as an effective way of evaluating both volumetric bone mineral density (vBMD) and geometric data in the femur. From Sep 2009 to Dec 2011, 296 postmenopausal women in their fifties who had experienced menopause within 5 years as well as the diagnosis of breast cancer itself were chosen. The subjects who received AIs (n=166, mean age 51.1[plusmn]0.3 y.o.) and the ones who did not get any adjuvant medication (n=130, mean age 51.9[plusmn]0.2 y.o.) had been examined by QCT at the time point of average 33.1[plusmn]1.1 months after the mastectomy. In addition to the measurement of axial volumetric bone mass, 2D BMD and hip geometry were analyzed by QCT PRO software (Mindways Software, Austin TX, USA). As the subjects were in their early menopausal period, there were no statistical differences in volumetric BMD of lumbar spine and femur, however, areal BMD in the femoral neck (FN) and total hip (TH) were significantly lower in patients who had been treated with AIs compared to non-AI treated group (FN: 0.656[plusmn]0.008 [italic]vs.[/italic] 0.691[plusmn]0.008 g/cm[sup]2[/sup], TH: 0.765[plusmn]0.008 [italic]vs.[/italic] 0.804[plusmn]0.011 g/cm[sup]2[/sup], P[lt]0.05, respectively). The cross-sectional area (CSA) and the average cortical thickness of FN were also significantly lower in AI-treated group (1.82[plusmn]0.03 [italic]vs.[/italic] 1.98[plusmn]0.45 cm[sup]2[/sup] 2.06[plusmn]0.05 [italic]vs.[/italic] 2.27[plusmn]0.54 mm, P[lt]0.05, respectively). There were no differences between two groups regarding angle, width, axis length and buckling ratio in the FN. The use of AIs resulted in unfavorable cortical geometry of FN as compared to women on non-AI therapy in postmenopausal breast cancer patients. This indicates that AIs, by inducing absolute deficiency in the estrogen level, lead to changes in the increased endocortical bone resorption with accordingly reduced cortical thickness in the FN much earlier than expected before the occurrence of the well-known bone loss in the spines. In conclusion, use of AIs in patients with breast cancer in their early menopausal period caused negative alteration in the cortical geometrical parameters which might increase the risk of osteoporotic fractures.[br][br]Nothing to Disclose: SJL, KMK, J-HS, S-KL, BWP, YR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1393 60 426 SAT-336 PO05-01 Saturday 365 2012


366 ENDO12L_SAT-337 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Association between Bone Mineral Density and Incidence of Breast Cancer in Southern Israel Merav Franekel, Yair Liel, Michael Koretz, Ethel Siris, Larry Norton, Tali Shafat, Shraga Shani, Victor Novack, David B Geffen Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel; Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel; Columbia University Medical Center, New York, NY; Memorial Sloan Kettering Cancer Center, New York, NY; Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel; Faculty of Health Sciences, Ben Gurion University of the Negev, Beer Sheva, Israel; Faculty of Health Sciences, Ben Gurion University of the Negev, Beer Sheva, Israel Introduction[br]Osteoporosis and breast cancer affect many postmenopausal women. Estrogen has a central role in the maintenance of bone integrity. Its deficiency leads to accelerated bone resorption and decreased BMD. Some studies have suggested an inverse relationship between BMD and the incidence of breast cancer. The primary objective of this study was to assess whether BMD is associated with risk of subsequent development of breast cancer in the female population of the southern Israel treated at Soroka University Medical Center (SMC).[br]Methods:[br]The electronic medical charts of women who underwent DEXA BMD studies at SMC between February 2003 and March 2011 were screened for breast cancer diagnosed after DEXA was performed, using ICD-9 codes. Demographic, BMD and laboratory findings were compared between those who were subsequently diagnosed with breast cancer and those who remained cancer-free. Women were divided into groups based on the tertiles of BMD at 3 sites: lumbar spine (LS,L1-4), total hip (TH) and neck of femur (FN). The incidence of breast cancer was calculated for each group.[br]Results:[br]Of 15268 women who underwent BMD testing, 807 had a previous diagnosis of breast cancer and 86 were diagnosed with breast cancer after BMD testing. The mean age of those diagnosed with breast cancer was higher than those without (68.8[plusmn]9.1 vs. 65.1[plusmn]11 years.; p[lt]0.001) and they had a higher BMI (30.9[plusmn]5.5 vs. 29.1[plusmn]5.7 p=0.004). The majority of women in both groups were older than 50 (94.2% and 92.7% p=0.597), and they did not differ in the level of vitamin D and PTH.[br]The mean Z-score BMD of breast cancer patients was significantly higher than controls in all 3 sites (LS: 0.36[plusmn]1.58 vs. -0.12[plusmn]1.42, p=0.002; TH: 0.371.08[plusmn] vs. 0.03[plusmn]1.02, p=0.002; FN: 0.04[plusmn]0.99 vs.-0.18[plusmn]0.94; p=0.026)[br]Women with higher Z scores at all three sites had a significantly higher chance of developing breast cancer with an odds ratio of 2.1, 2.2 and 1.85 for the FN, TH and LS(per tertile, p value 0.006, 0.005 and 0.03 respectively). No correlation was found between the BMD Z-score and the stage, histology, grade or survival from breast cancer.[br]Conclusions[br]This study confirms previously published data on a direct association between BMD and the risk for breast cancer. The limitation of this study is the lack of information about several other confounders that may have affected BMD and breast cancer risk such as age of menarche and menopause, gravidity, parity, HRT usage and anti resorptive medications.[br][br]Sources of Research Support: USA Breast Cancer Foundation, unrestricted research grant.[br][br]Nothing to Disclose: MF, YL, MK, ES, LN, TS, SS, VN, DBG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 305 60 427 SAT-337 PO05-01 Saturday 366 2012


367 ENDO12L_SAT-338 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) The Effects of Telmisartan on Bone Turnover Markers Emrah Eraslan, Sevim Gullu Ankara University, Faculty of Medicine, Ankara, Turkey; Ankara University, Faculty of Medicine, Ankara, Turkey Introduction and Purpose: It is known that a group of oral hypoglycemic drugs, thiazolidinedions, are agonists for PPAR[gamma] receptors and have negative effects on bone metabolism. And there are some data about beneficial effects of RAS blocking on bone metabolism. But the effect of telmisartan, which is an Ang II AT1 receptor blocker and partially agonist for PPAR[gamma], on bone metabolism is not known. The aim of the study was to determine the total effect of telmisartan use on bone metabolism.[br]Materials and Methods: The patients who have newly diagnosed as stage I hypertension were enrolled and randomised to telmisartan (N=24) or losartan (N=21) group. The patients who had bone diseases and disease or drug use which may affect bone metabolism were excluded. Serum Ca, P, 25 OHD, bone alkaline phosphatase (BALP), osteocalcin (OC), IL-6 and 24 hours urinary NTx levels were measured before the start of treatment and at third month visit.[br]Findings: At the end of three months period Ca and P levels did not change. But statistically significant increase in the levels of 25 OHD [15.4 (10.2-41.9) vs. 17.8 (10.4-49.1) [micro]g/mL, p=0.010], and statistically significant decreases in the levels of PTH (54.39[plusmn]25.98 vs. 45.12[plusmn]22.87 pg/mL, p[lt]0.001), BALP (16.51[plusmn]5.02 vs. 15.06[plusmn]4.06 [micro]g/L, p=0.008), OC (11.27[plusmn]5.03 vs. 8.96[plusmn]5.65 ng/mL, p=0.045), urinary NTx (96.37[plusmn]64.51 vs. 43.08[plusmn]35.68 nM BCE/mM, p[lt]0.001), IL-6 [17.25 (2.30-196.9) vs. 9.5 (0.8-137.6) pg/mL (p=0.002)] were observed. There were no differences between the two groups regarding the changes of these parameters. After the covariant analysis which was made for eliminate of 25 OHD effect; there were no significant changes at any of the parameters and partial correlation analysis showed a positive correlation between NTx and IL-6 (R=0.338, p=0.033). After the elimination of PTH effect the positive correlation of NTx between IL-6 was steady in partial correlation analysis (R=0.382, p=0.015).[br]Conclusion: The use of telmisartan in humans does not affect the bone metabolism despite its partial PPAR-gamma agonistic effect.[br][br]Nothing to Disclose: EE, SG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1203 60 428 SAT-338 PO05-01 Saturday 367 2012


368 ENDO12L_SAT-339 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Use of the FRAX Calculator with and without Bone Mineral Density (BMD) in Greek Women Ioannis Ilias, Filio Spanoudi, Maria Alexiou, Eftychia Koukkou, Stamatina C Nikopoulou E Venizelou Hospital, Athens, Greece Introduction[br]The creators of the FRAX fracture risk calculator for countries with no national reference values (such as Greece) suggest the use of FRAX from a neighboring country. For Greeks (considered to run a high risk for osteoporotic fractures) the Italian FRAX is the suggested choice, although endocrinologists also use the Turkish FRAX. The correspondence of FRAX results with and without the use of femoral neck T-scores with DXA is considered to be adequate.[br]Aim[br]To assess the correspondence of 10-year femoral neck fracture risk with and without the inclusion of femoral neck T-scores implementing the Italian or Turkish FRAX in Greek women and to evaluate [ndash] albeit indirectly - its validity.[br]Subjects[br]We studied 98 consecutive women outpatients (mean age+SD: 58+8 years, BMI+SD:26+4 kg/m2) with 0-3 FRAX risk factors. All had neck femoral BMD measured with DXA and received only calcium supplements+vitamin D3; all were bisphosphonate-naive. We calculated the 10-year femoral neck fracture risk with and without inclusion of femoral neck T-scores, drafted Bland-Altman diagrams and calculated Cohen[apos]s Kappa coefficient (with the accepted 3% 10-year femoral neck fracture risk as a discriminatory threshold).[br]Results[br]Based on DXA measurements 23 women had BMD within normal limits, 54 had osteopenia and 21 had various degrees of osteoporosis. The Bland-Altman diagrams showed that although the correspondence of FRAX scores was acceptable for most women there were nevertheless inconsistencies for a substantial number of them. Discrepancies in FRAX scores with clinical importance were noted in 22/98 (Kappa=0.34) and 24/98 (Kappa=0.24) women with the Italian or Turkish FRAX, respectively.[br]Conclusion[br]The correspondence of femoral neck fracture risk in Greek women using the Italian version of FRAX (with and without the inclusion of femoral neck T-scores) was not satisfactory (78%) and was worse than the one reported during the validation of FRAX in other countries (approximately 85%-90%). Correspondence of values with the Turkish FRAX was slightly lower (75%). We have to note that the validity of the Turkish FRAX has been questioned (1). Our results are important in view of the proposed use of FRAX as a means to identify low- or high-fracture risk patients that do not require DXA and has to be taken into consideration when interpreting results in Greek women. A Greek FRAX seems necessary.[br][br](1) Tuzun et al, Osteoporos Int. 2011; DOI: 10.1007/s00198-011-1655-5.[br][br]Nothing to Disclose: II, FS, MA, EK, SCN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 940 60 429 SAT-339 PO05-01 Saturday 368 2012


369 ENDO12L_SAT-340 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) The Effect of Bisphosphonate Treatment on Histomorphometric and Clinical Indices in Adolescents with Idiopathic Juvenile Osteoporosis Jennifer Harrington, Etienne Sochett The Hospital for Sick Children, Toronto, Canada [bold]Context:[/bold]Idiopathic juvenile osteoporosis (IJO) is an uncommon condition in children and adolescents, characterized by frequent fractures and significant bone pain in the absence of underlying systemic disease or evidence of a collagen disorder. The exact pathogenesis and optimal management strategy in IJO is still unclear. Bisphosphonates have been shown to reduce bone pain but the effect on bone histomorphometry in IJO is unknown.[br][bold]Objectives:[/bold]To assess the clinical and histomorphometric data in a cohort of children and adolescents with IJO, and to compare these findings in those who had received treatment with bisphosphonates with those who had not received any specific pharmacotherapy at the time of bone biopsy.[br][bold]Results:[/bold]21 patients (12 males) with IJO, aged 13.2([plusmn]3.0) years at the time of transiliac bone biopsy were assessed. None had a history of a secondary systemic disease or had abnormalities of sclera, joint hypermobility or skin laxity. Median number of previous peripheral fractures was 3(0-8) and vertebral fracture was 2(0-8), and all had a significant history of bone pain. The cohort had a mean height Z score of -0.4([plusmn]1.2), weight Z score of -0.1 ([plusmn]1.2) and BMI Z of 0.1(1.3). Median 25-hydroxy Vitamin D was 77(21-127) nmol/L with 3 subjects having levels [lt] 50 nmol/L.[br]8 (6 males) patients had previously been treated with bisphosphonates for a mean of 2.3([plusmn]1.3) years. Lumbar spine BMD in the treated group significantly improved post treatment (Z score -2.8 to -1.7, p=0.009) but was not different at time of bone biopsy from the untreated group (Z score -2.0). In the group overall, on histomorphometry 11(52%) subjects had evidence of low bone volume (BV/TV[lt] -1SD) and trabecular thickness was subnormal in 8(38%). Bone volume positively correlated with age (r=0.43, p=0.04). There was a trend toward less significant impairment in bone volume in the treated group compared to the untreated group (BV/TV -1.3 vs. -0.11, p=0.09). Of those who had tetracycline labeling, all of the untreated patients had evidence of normal or high bone turnover (n=8/13) compared to low turnover in all treated patients (n=6/8).[br][bold]Conclusions:[/bold]Bisphosphonate use in adolescents with IJO is associated with increased BMD Z scores and bone volume as measured by histomorphometry. Caution may be needed with its use in IJO given the presence of uniform low turnover in treated patients. Larger prospective studies are needed to assess the impact of bisphosphonates in patients with IJO.[br][br]Nothing to Disclose: JH, ES 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1839 60 430 SAT-340 PO05-01 Saturday 369 2012


370 ENDO12L_SAT-341 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Low Bone Density and Effects of Alendronate Therapy in Patients with Duchenne Muscular Dystrophy Caroline Houston, Amal Shibli-Rahhal University of Iowa Hospitals and Clinics, Iowa City, IA [bold]Background: [/bold]Duchenne Muscular Dystrophy (DMD) is characterized by progressive muscle weakness that leads to loss of mobility by age 10 and death by age 20. Glucocorticoids are the mainstay of treatment, as they have been shown to slow disease progression. DMD patients develop low bone density as a result of glucocorticoid use and loss of mobility; however, little data exists to guide efforts towards screening for and treating low bone density in this population. We examined changes in bone density and the effects of alendronate therapy in DMD patients treated at the University of Iowa in the last decade.[br][bold]Methods: [/bold]We conducted a retrospective cohort study of 39 patients with DMD for whom bone density data was available between December 2000 and September 2011. We examined baseline patient characteristics and bone density. We then assessed changes in bone density with alendronate therapy.[br][bold]Results: [/bold]The mean patient age at the time of diagnosis was 3.5 years. Mean follow-up was 15.1 years. Ninety percent became wheelchair bound at a mean age of 10.6 years. Seventy-four percent were treated with glucocorticoids for an average period of 8.4 years. Seventy-two percent were treated with alendronate for an average period of 3.8 years.[br]Patients received their first DXA scan an average of 8.5 years after diagnosis. Their bone density was low for their age, and the mean z-score was lower at the hip than at the lumbar spine (-3.27 versus -1.84, p [lt] 0.0001). Patients treated with glucocorticoids received their first DXA scan sooner after diagnosis (7.7 versus 10.9 years), yet had lower z-scores at the lumbar spine (-2.04 versus -1.12, p = 0.0014). Among 8 patients who received multiple DXA scans prior to starting alendronate, the mean z-score at the hip declined from -2.85 to -3.04 (p = 0.70) over an average of 4.0 years. Among 18 patients treated with alendronate for an average of 3.0 years, mean z-scores increased at the hip (-3.31 to -3.25, p = 0.836) and lumbar spine (-2.09 to -1.80, p = 0.523).[br][bold]Conclusions: [/bold]Patients with DMD develop low bone density that is more severe at the hip than the lumbar spine. Our patients were found to have low bone density 8.5 years after diagnosis and may have benefitted from earlier screening. Z-scores at the hip tended to decline with time prior to alendronate therapy. Z-scores at the hip and lumbar spine tended to improve with alendronate therapy. These trends were non-significant, likely due to small sample size.[br][br]Nothing to Disclose: CH, AS-R 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 441 60 431 SAT-341 PO05-01 Saturday 370 2012


371 ENDO12L_SAT-342 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Spine Bone Mineral Density (BMD) in Children with Duchenne Muscular Dystrophy Treated with Corticosteroids Gisella Viterbo, Cristina Tau, Soledad Monges, Juliana Catagneto, Alicia Belgorosky Hospital de Pediatria Garrahan, Buenos Aires, Argentina; Hospital de Pediatria Garrahan, Buenos Aires, Argentina Reduced mobility and glucocorticoids as adjunctive therapy may cause osteoporosis and fractures in children with Duchenne muscular dystrophy (DMD). We analyzed in 21 boys (mean age [plusmn] SD: 11.6 [plusmn]2.7y, weight SDS: -0.27[plusmn]1.16 (-2.70 to 2.65), and height SDS: -1.77[plusmn]0.87) DMD after treatment with deflazacort (n=13) or methylprednisone (n=8) during 3.8[plusmn]2.4 years (range, 0.4-8.75), vitamin D (300 to 2400 IU/day) and calcium supplement (0.13 to 1 g/day) and calcium intake by dairy products of 607[plusmn]231 mg/day: Lumbar L2-L4 BMD measured by dual-energy X-ray absorptiometry (Lunar, Prodigy), serum calcium (Ca), phosphate (P), alkaline phosphatase (AP), PTH, urinary calcium and D-Pyridoline/creatinine (uD-Pyr). Ten patients were wheelchair-bound (mean age: 13.2 [plusmn]2.3 y, immobilization time was 2.04 [plusmn]1.9 y) while long bone (n=3) and vertebral crush fractures (n=7) patients were also observed. The cumulative corticosteroid dose was 27.5[plusmn]19.8 g (range: 4.7-65.7). Mean BMD z score was -2.2[plusmn]1.5, ranging from -5.9 to 0.4. BMD z score was [lt]-2 in 43%, and between -1 and -2 in 38%. Mean BMD z score was inversely correlated with age (p[lt]0.001), time of immobilization (p[lt]0.001), duration of corticosteroid therapy (p[lt]0.01) and cumulative dose of corticosteroids (p[lt]0.001). Significantly lower BMD z score (-3.78[plusmn]1.26) was observed in patients with, in comparison with those without vertebral fractures (-1.43[plusmn]0.62, p[lt]0.005). The number of fractures was positively correlated with cumulative corticosteroids dose. Ca, P, AP and PTH were within the normal range. Urinary calcium and uD-Pyr were increased in 38 % and 56 % of the patients, respectively. In conclusion long-term immobilization and treatment with high corticoid doses affect bone mineralization in children with DMD and might worsen the outcome of the disease.[br][br]Nothing to Disclose: GV, CT, SM, JC, AB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 467 60 432 SAT-342 PO05-01 Saturday 371 2012


372 ENDO12L_SAT-343 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) The Evaluation of Teriparatide and Alendronate in Cystic Fibrosis Oranan Siwamogsatham, Vin Tangpricha Emory University Hospital, Atlanta, GA Introduction: Cystic fibrosis (CF) is a common genetic disease that has effects on multiple organs, including bone. The major contributing factors of bone disease in cystic fibrosis include vitamin D malabsorption, poor nutritional status, physical inactivity, chronic inflammation, glucocorticoid therapy, delayed puberty and hypogonadism. These factors can lead to a reduction in bone density, osteoporosis, fragility fractures and kyphosis. There is limited long term data regarding the effectiveness of osteoporosis treatments in these patients. The objective of our study was to examine the efficacy of teriparatide and alendronate in adult subjects with CF and low bone density.[br]Methods: We conducted a retrospective chart review of all CF patients who were diagnosed as having osteoporosis and received teriparatide or alendronate treatment at the Emory CF program during 2006-2011. The study was approved by the IRB at Emory. We collected demographic information, bone mineral density (BMD) at baseline and 2 years after treatment, and all adverse events.[br]Results: We identified 11 subjects with CF who were treated with osteoporosis medication during this study period. Six were treated with teriparatide and 5 with alendronate. Both groups were comparable in demographic information except subjects receiving teriparatide had significantly lower BMI (p=0.02). The BMD was similar in both groups at the lumbar spine, left total hip and femoral neck. After 2 years of treatment, the group receiving teriparatide had significant improvement in the left femoral neck (0.711[plusmn]0.096 to 0.762[plusmn]0.012 g/cm2, p=0.02). We did not detect a significant increase in BMD at any site the group receiving alendronate, likely due to the small sample size. No adverse events were detected in either group.[br]Conclusion: In patients with CF, therapy with teriparatide appears to be effective. Our sample size precludes any conclusion to whether teriparatide is superior to alendronate in the treatment of CF bone disease. Large and larger term studies are required to examine the optimal therapy for CF bone disease.[br][br]Nothing to Disclose: OS, VT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 953 60 433 SAT-343 PO05-01 Saturday 372 2012


373 ENDO12L_SAT-344 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Factors Related to Decreased Bone Mineral Density in Childhood Cancer Survivors Yun Jung Choi, Moon Hee Lee, Won Kyoung Cho, Kyoung Soon Cho, So Hyun Park, Min Ho Jung, Byung Kyu Suh, Byung Churl Lee College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea [bold]Purpose[/bold]: The risk of osteoporosis or osteopenia is known to increase after childhood cancer treatment. The purpose of this study was to evaluate patterns of bone mineral density (BMD) and factors related to decreased BMD in childhood cancer survivors.[br][bold]Method[/bold]: We studied 78 patients (34 males, 44 females) treated for acute lymphoblastic leukemia, acute myelogenous leukemia or chronic myelogenous leukemia during childhood. Clinical data, laboratory finding, bone age, lumbar BMD (LBMD) and femur neck BMD (FNBMD) were investigated. Chronological age at evaluation was 11.6[plusmn]3.4 yr in males and 13.0[plusmn]3.3 yr in females. Primary disease, chronological age at treatment, body mass index (BMI), method of treatment (chemotherapy, radiotherapy), endocrine function, presence of chronic graft-versus-host disease (cGVHD) and relevance of BMD standard deviation score (SDS) were evaluated.[br][bold]Results[/bold]: LBMD and FNBMD of the subjects were -0.91[plusmn]1.41 and -1.13[plusmn]1.79, respectively. Twenty (25.7%) patients had LBMD SDS lower than -2. Nineteen (24.4%) patients had FNBMD SDS lower than -2. The patients treated with hematopoietic stem cell transplantation had lower LBMD SDS (-1.17[plusmn]1.39 vs -0.43[plusmn]1.33, [italic]P[/italic]=0.025). The risk of having LBMD SDS [lt]-2 was higher in the patients treated with glucocorticoid for cGVHD (36.6% vs 13.5%; odds ratio [OR], 3.7; [italic]P[/italic]=0.020). In multivariate logistic regression analysis, longer duration of glucocorticoid treatment (OR, 1.12; 95% confidence interval [CI], 1.03-1.22) and lower BMI SDS (OR, 0.42; 95% CI, 0.21-0.83) were associated with decreased LBMD SDS.[br][bold]Conclusions[/bold]: These findings suggest that prolonged glucocorticoid use and reduction in BMI are risk factors for decreased BMD in childhood cancer survivors. Anticipatory follow-up and appropriate treatment are necessary, especially for the patients with risk factors.[br][br]Nothing to Disclose: YJC, MHL, WKC, KSC, SHP, MHJ, BKS, BCL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 786 60 434 SAT-344 PO05-01 Saturday 373 2012


374 ENDO12L_SAT-345 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Spooning Pattern of Age-Related Changes of Femoral Bone Mineral Density in Youth: From a Nationwide Study (KNHANES IV) Eun Young Lee, Daham Kim, Kyoung Min Kim, Kwang Joon Kim, Han Seok Choi, Yumie Rhee, Sung-Kil Lim Yonsei University College of Medicine, Seoul, Korea; Severance Hospital, Seoul, Korea; Dongguk University College of Medicine, Ilsan, Korea [bold]Context:[/bold] Bone loss has been considered to begin with menopause in women and later in life in men. Recently, several studies reported that bone loss began in young adults. There are still discordant results concerning age-related changes in bone mineral density (BMD), especially in non-vertebral bone.[br][bold]Objective:[/bold] To investigate the age-related changes in BMD in Korean youth.[br][bold]Design and Setting:[/bold] This was a population-based, cross-sectional study from the Fourth Korea National Health and Nutrition Examination Surveys (KNHANES IV).[br][bold]Participants:[/bold] A total 10,575 Korean (men 4,731 and women 5,844) aged 10 to 80 yr were included.[br][bold]Main outcome Measures:[/bold] BMD at spine and hip was measured using dual X-ray absorptiometry.[br][bold]Results:[/bold] Age-related bone loss at femoral neck in men occurred continuously with temporary acceleration phase after achieving peak bone mass (PBM). In contrast, age-related bone loss at total hip in both genders and femoral neck in women showed obvious 3 phases; acceleration, consolidation, and then the second acceleration phase after reaching PBM. Interestingly, this pattern of bone loss was more significant in total hip, thus, showed acceleration phase until late 20[apos]s and consolidation phase until late 40[apos]s. Early accelerated loss of BMD was not observed at lumbar spine in both genders. Although body mass index and body fat percent were more related with BMD than other clinical parameters, they could not explain the early accelerated loss of BMD at femur.[br][bold]Conclusions:[/bold] There was accelerated bone loss at femur in both genders during early young adult life and more than 60% of bone loss before age 50 yr occurred during this period.[br][br]Sources of Research Support: This work was supported by the National Research Foundation (NRF) of Korea grant funded by the Korea government (MEST) (No. 20110001024).[br][br]Nothing to Disclose: EYL, DK, KMK, KJK, HSC, YR, S-KL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1448 60 435 SAT-345 PO05-01 Saturday 374 2012


375 ENDO12L_SAT-346 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Improving the Bone Health of Adolescent Patients with Hormonal Deficiencies Pamela Corlett, Pauline Whittingham, Aftab Ahmad The Royal Liverpool and Broadgreen University Hospital, Liverpool, UK Introduction[br]Evidence suggests bone mineral density is reduced in adolescent patients with hormonal deficiencies. Consensus agreement recommends the management of adolescent patient transitioning to adult services be conducted through specialist clinics. These Transitional clinics should be staffed by a multi-disciplinary team experienced in managing hypopituitarism and growth hormone deficiency.[br]We established our Transitional Clinic in 2007, staffed by Consultant Paediatric Endocrinologist, Consultant Endocrinologist (Adult) and Endocrine Specialist Nurse.[br][bold]Method:[/bold] we performed a retrospective audit of 42 patients attending our Transitional Clinic, using patient casenotes and hospital systems. All patients had undergone DEXA scanning of the lumbar spine and left femoral neck.[br]Demographic[br]A total of 42 patient records were accessed, of these 29 (69%) had a low BMD. Of the 29 patients with low BMD Brain tumour/injury (n=10) Histocytosis(n=2), Leukaemia (n=3), CAH (n=1), BPS (n =)1, Primary hypogonadism (n=1), Thallasaemia (n=1) Hypogonadotrophic hypogonadism (n=1)[br]Treatment[br]Of the 29 with low BMD 17/29 received combination of bisphosphonate and calcium/vit d, whilst 3 received calcium/vit d only [amp] 9 patients were untreated. In the group treated with bisphosphonate and calcium 8 (47%) showed improved BMD when rescanned, 3 had no change and 2 showed reduction in BMD while 4 await follow up scans. Of those treated with Calcium/Vit d 1 (33%) improved at rescan and 2 await follow up scans. In the not treated group 1 showed improvement at rescan and 1 showed no change while 7 await follow up scans[br]Of the 29 with low BMD 16 (55%) had childhood growth hormone deficiency, 2 of which remained on growth hormone replacement. 10/16 received bisphosphonate+calcium of which 4 (40%) showed improved BMD, 3 (30%) no change and 3 await rescan. 2/16 received Calcium/VitD both of which are awaiting rescan [amp] 4 received no treatment awaiting rescans.[br]Conclusion[br]Low BMD is present in 2/3 of patients attending our Transitional clinic. In those treated with combined bisphosphonate and calcium vitamin d 47% showed improvement in BMD when rescanned whereas a further 18% had stable BMD on repeat scanning. Further monitoring is needed to assess optimum treatment duration for continued bone health.[br][br]Nothing to Disclose: PC, PW, AA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1210 60 436 SAT-346 PO05-01 Saturday 375 2012


376 ENDO12L_SAT-347 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) The Markers of Bone Metabolism and Bone Quality in Patients with Anorexia Nervosa Ayako Urano, Mari Hotta, Mariko Araki, Rina Ohwada, Atsuhiro Ichihara Tokyo Women[apos]s Medical University, Tokyo, Japan; National Graduate Institute for Policy Studies, Tokyo, Japan Introduction [amp] Objective: Anorexia nervosa (AN) is frequently accompanied by osteoporosis. They have multiple risk factors for osteoporosis such as low body weight, malnutrition, a decrease in serum insulin-like growth factor-I (IGF-I) as an osteogenic factor, and estrogen deficiency. Recently, bone strength is evaluated by both bone mineral density (BMD) and bone quality. To evaluate bone strength, we investigated BMD, bone metabolism, and bone quality in AN patients. Subjects [amp] Methods: Fifty-five female AN patients (restricting type: 32, binge-eating/purging type: 24, age: 26.1 [plusmn] 5.6 (mean [plusmn] SD) years, body mass index (BMI): 14.8 [plusmn] 2.6 kg/m[sup]2[/sup]) and Tweleve age-matched healthy female (28.8 [plusmn] 3.3 years, 21.1 [plusmn] 3.1 kg/m[sup]2[/sup]) were included in this study. We measured nutritional factors including of 25-hydroxy-vitaminD (25OHD),1,25-dihydroxy-vitaminD (1,25OHD), and undercalboxylated osteocalcin (ucOC), serum IGF-I, estradiol(E2), intact PTH, leptin, and adiponectin, bone specific alkaline phosphatase, osteocalcin (OC) as bone formation markers, urinary bone resorption markers of N-terminal telopeptide (NTX) and carboxyterminal telopeptide (CTX) of type I collagen, and homocysteine and pentosidine as bone quality markers. Results: In AN patients, lumbar BMD measured by dual-energy X-ray absorptiometry was extremely low (T-score: -2.2 [plusmn] 1.1), but there was no significant difference between restricting type and binge-eating/purging type. Serum levels of IGF-1, E2 were significantly low, while urinary NTX was significantly high. Serum levels of ucOC was elevated in 43% of AN patients, which indicated deficiency of vitamin K intake. In 61% of AN patients, serum levels of 25-OHD was lower than 20 pg/ml, which indicated deficiency of vitamin D intake. While bone quality markers of AN patients under 30 years of age were within normal range, both markers of those over 30 years of age significantly increased, which indicated deterioration of bone quality. Discussion [amp] Conclusion: Since a considerable number of AN patients have a lack of vitamin D as well as vitamin K, those vitamins would be available for prevention or treatment of osteoporosis. We have to pay attention to bone quality in AN patients aged as early as 30 years.[br][br]Nothing to Disclose: AU, MH, MA, RO, AI 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1078 60 437 SAT-347 PO05-01 Saturday 376 2012


377 ENDO12L_SAT-348 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Serum Sodium Levels Are Predictive of Bone Loss Severity in Anorexia Nervosa Elizabeth A Lawson, Pouneh Fazeli, Genevieve Calder, Hannah Putnam, Madhusmita Misra, Erinne Meenaghan, Karen K Miller, Anne Klibanski Massachusetts General Hospital and Harvard Medical School, Boston, MA Anorexia nervosa (AN) is a psychiatric disorder characterized by restrictive eating, low body weight, and severe bone loss. In studies of young women with AN, the vast majority have osteopenia and up to 40% meet WHO criteria for osteoporosis at one or more sites. Recent data show a deleterious relationship between low serum sodium levels and bone mass, and relative or absolute hyponatremia is a known complication of AN. Verbalis et al. demonstrated that experimentally induced hyponatremia in rats results in a marked reduction in bone mineral density (BMD) and trabecular and cortical parameters. Clinical studies of other medical conditions associated with hyponatremia suggest that detrimental effects of low sodium levels on health are seen even within the normal range. We hypothesized that women with AN and relatively low sodium levels would have lower BMD than those with higher sodium levels. In a cross-sectional study of 404 women ages 17-54 (mean[plusmn]SEM age 25.6[plusmn]0.3 years) who met DSM-IV criteria for AN, we measured BMD using DXA. BMD was compared in women with sodium levels [lt]140mmol/L (midpoint of normal range) (N=114) vs. those with sodium levels [ge]140mmol/L (N=287), and in women with hyponatremia (sodium[lt]135mmol/L) (N=12) vs. those without (N=389) using Student[apos]s T-test. Multivariable least-squares analyses were constructed to control for potential confounders. Women with sodium levels [lt]140mmol/L had lower BMD, T- and Z-scores at the AP spine and total hip vs. those with sodium levels [ge]140mmol/L (T-scores: AP spine -1.8[plusmn]0.1 vs. -1.4[plusmn]0.1, hip -1.2[plusmn]0.1 vs. -0.9[plusmn]0.1, p[lt]0.01). Differences remained significant after controlling for age, BMI, and psychiatric medications, including serotonin reuptake inhibitors (SSRI[apos]s). After controlling for disease duration, differences at the AP spine remained significant. Hyponatremic women had lower BMD, T- and Z-scores at the AP spine, lateral spine and total hip vs. those without hyponatremia (T-scores: AP spine -2.6[plusmn]0.4 vs. -1.5[plusmn]0.0, lateral spine -3.2[plusmn]0.5 vs. -1.8[plusmn]0.1, hip -2.6[plusmn]0.5 vs. -1.0[plusmn]0.1, p[lt]0.003). Differences remained significant after controlling for age, BMI, disease duration, and psychiatric medications, including SSRI[apos]s. These data suggest that relative sodium deficiency may contribute to AN-related osteopenia.[br][br]Sources of Research Support: National Institutes of Health Grants R01 DK052625, DK062249,. DK084970, MH083657, ULIRR0257801 and M01 RR01066.[br][br]Nothing to Disclose: EAL, PF, GC, HP, MM, EM, KKM, AK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 948 60 438 SAT-348 PO05-01 Saturday 377 2012


378 ENDO12L_SAT-349 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Obese Male Adolescents Have an Increased Cross-Sectional Bone Size and Volumetric Trabecular Bone Mineral Density at the Tibia Sara Vandewalle, Inge Roggen, Patrick Debode, Maria van Helvoirt, Greet Roef, Eva Van Caenegem, Youri Taes, Jean De Schepper, Jean-Marc Kaufman University Hospital Ghent, Ghent, Belgium; University Hospital Ghent, Ghent, Belgium; University Hospital Brussels, Brussels, Belgium; Zeepreventorium, De Haan, Belgium [bold]Context: [/bold]A growing concern exits that childhood obesity may negatively affect bone development, because of increasing evidence linking childhood obesity to skeletal fractures. Whereas some studies report greater bone mass (measured by DXA) in overweight children, other studies using DXA conclude that obesity is linked to a lower bone mineral density (BMD). There is still limited information about the true volumetric BMD (vBMD) of long bones.[br][bold]Objective: [/bold]This study aims to determine whether obese adolescents (ObA) are at risk for a lower vBMD and altered bone geometry at the level of the tibia.[br][bold]Methods: [/bold]51 male ObA (mean BMI sds: 2.55), aged 10-19y, at start of a residential obesity treatment program and 51 healthy (mean BMI sds: -0.17) controls, matched for age, height and pubertal status were included in a cross-sectional study. Trabecular (at the distal 4% end) and cortical (at the 66% site) vBMD and cross-sectional geometry were assessed at the non-dominant tibia using peripheral quantitative computed tomography. Whole body lean- and fat mass were determined using DXA.[br][bold]Results: [/bold]Compared to the controls, ObA had not only a higher fat mass (44.9 [plusmn] 12.4 vs. 8.2 [plusmn] 4.0 kg; p [lt]0.001), but also a higher lean mass (51.2 [plusmn] 13.0 vs. 42.6 [plusmn] 10.0 kg; p [lt]0.001). Trabecular bone content (131 [plusmn] 35 vs. 103 [plusmn] 30 mg/cm; p [lt]0.001), vBMD (240 [plusmn] 29 vs. 227 [plusmn] 35 mg/cm[sup3]; p = 0.04), as well as bone area (545 [plusmn] 111 vs. 452 [plusmn] 97 mm[sup2]; p[lt]0.001) at the distal tibia were higher in ObA versus the controls. At the mid shaft, both cortical bone content (332 [plusmn] 81 vs. 278 [plusmn] 64 mg/cm; p [lt]0.001) and area (307 [plusmn] 69 vs. 257 [plusmn] 51 mm[sup2]; p[lt]0.001), as well as the periosteal (80 [plusmn] 10 vs. 74 [plusmn] 8 mm; p [lt]0.001) and endosteal (52 [plusmn] 8 vs. 46 [plusmn] 10 mm; p =0.004) circumferences were larger in the obese group. There was no significant difference in cortical thickness and cortical vBMD between the groups. In the merged study groups significant positive associations were observed between body weight and cortical bone area (p[lt]0.001), periosteal circumference (p[lt]0.001), endosteal circumference (p[lt]0.001) trabecular bone area (p[lt]0.001) and trabecular vBMD (p=0.01), but not cortical vBMD.[br][bold]Conclusion:[/bold]During adolescence obesity results in a greater cross-sectional tibial bone size, probably resulting from an accelerated periosteal apposition, related to a greater mechanical load with increased muscle mass and force. Our data do not support the hypothesis that the higher fracture rate in ObA is caused by a lower vBMD.[br][br]Sources of Research Support: SV and YT are holders of a respectively a PhD fellowship and postdoctoral fellowship, from the Research Foundation, Flanders (FWO).[br][br]Nothing to Disclose: SV, IR, PD, MvH, GR, EVC, YT, JDS, J-MK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1108 60 439 SAT-349 PO05-01 Saturday 378 2012


379 ENDO12L_SAT-350 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Hormonal Determinants of Bone Microarchitecture in Obese Men Miriam A Bredella, Eleanor Lin, Anu V Gerweck, Karen K Miller Massachusetts General Hospital and Harvard Medical School, Boston, MA; Massachusetts General Hospital and Harvard Medical School, Boston, MA Context: Obesity in men is associated with dysregulation of the GH/IGF-1 and gonadal steroid axes, important regulators of bone homeostasis. Studies suggest that obesity is detrimental to bone health. It is unknown whether endocrine dysregulation impacts skeletal microarchitecture in obese men.[br]Objective: To investigate endocrine determinants of bone microarchitecture in obese men.[br]Participants: 35 obese men (mean age: 33.7[plusmn]6.4 y, mean BMI: 36.5[plusmn]5.8 kg/m[sup]2[/sup]).[br]Outcome measures: Distal radius trabecular and cortical microarchitecture by 3D hr-pQCT: average bone density (D100), trabecular to tissue volume (BV/TV), trabecular density (Dtrab), trabecular number (TbN), Tb thickness (TbTh), cortical area (CtAr); total and free estradiol and testosterone; IGF-1; peak glucagon-stimulated GH.[br]Results: Mean testosterone was 380 ng/dl (subject range 147-697 ng/dl; normal range 249-836 ng/dl). Mean estradiol was 34.5 pg/ml (subject range 15-73 pg/ml; normal range 1-44 pg/ml). Mean peak stimulated GH was 5.1 ng/ml (subject range 0.02-30.0 ng/ml), and mean IGF-1 SDS was [minus]1.8 (subject range [minus]2.6- [minus]0.7). Total (r=0.33, p=0.05) and free estradiol (r=0.34, p=0.05) were positively associated with D100, and there was a trend toward a positive association with Dtrab (r= 0.30, p=0.08), BV/TV (r=0.30, p=0.08), and TbTh (r=0.29, p=0.09). Total (r=0.33, p=0.05) and free (r=0.34, p=0.05) testosterone were positively associated with TbTh and inversely with TbN (r= [minus]0.33, p=0.05). Peak stimulated GH was positively associated with TbTh (r=0.43, p=0.01). IGF-1 was positively associated with CtAr (r=0.36, p=0.04). When TbTh was entered as a dependent variable and peak GH, IGF-1, estradiol and testosterone as independent variables in a stepwise regression model, only peak GH and estradiol were significant predictors of TbTh (p=0.02 and p=0.04, respectively); GH explained 18% and estradiol 11% of the variability of TbTh.[br]Conclusion: Testosterone, estradiol and GH are positive determinants of trabecular microarchitecture, and IGF-1 is a positive determinant of cortical microarchitecture in obese men. This suggests that decreased GH and testosterone, characteristic of male obesity, may exert deleterious effects on skeletal microarchitecture, whereas higher estradiol levels may be protective. Estradiol may be a more important determinant of bone microarchitecture than testosterone. The potential differential effects of GH and IGF-1 on trabecular vs. cortical microarchitecture warrant further study.[br][br]Sources of Research Support: In part by National Institutes of Health Grants R01 HL-077674, UL1 RR-025758, and K23 RR-23090.[br][br]Nothing to Disclose: MAB, EL, AVG, KKM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 374 60 440 SAT-350 PO05-01 Saturday 379 2012


380 ENDO12L_SAT-351 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Intrabdominal Visceral Adipose Tissue Is Negatively Associated with Lumbar Spine Volumetric Bone Mineral Density in Older Singapore Chinese Men Gilbert Wei-Kiat Ng, Adoree Yi-Ying Lim, Ling Ling Chan, Kavita Venkataramen, E Shyong Tai, Alvin Choong-Meng Ng Singapore General Hopsital, Singapore, Singapore; Singapore General Hopsital, Singapore, Singapore; National University Hospital Singapore, Singapore, Singapore; National University Hospital Singapore, Singapore, Singapore [bold]Background [/bold][br]Obesity is traditionally thought to be protective against osteoporosis. However, recent studies have suggested that increased fat mass is a risk factor for osteoporosis(1, 2). The relative importance of different fat depots on bone is not well studied.[br][bold]Methods[/bold][br]This is a cross-sectional observational study involving 57 healthy Chinese male age 60-80 years. Computed tomography (CT) was used to determine visceral adipose tissue (VAT) and subcutaneous tissue (SAT) at L2/3 vertebral level and volumetric bone mineral density (vBMD) at the lumbar spine (LS), femoral neck (FN) and total hip (TH).[br][bold]Results[/bold][br]Separate linear regression analyses showed that VAT was negatively associated with LS vBMD (p[lt].05), after adjusting for age, height, and weight. This negative association between was not significantly modulated by BMI classification (obesity [gt] 25 kg/m2). No significant associations were found between VAT with FN vBMD and TH vBMD, although there was a trend towards an inverse relationship at these sites. No significant associations were found between SAT and vBMD at all three sites.[br][bold]Discussion [/bold][br]Our findings suggest that VAT may have a detrimental impact on bone mass, particularly in the lumbar spine in elderly Chinese men. This warrants further investigations to elucidate the underlying pathophysiologic mechanisms which may yield new therapeutic targets for osteoporosis.[br][br]1. Hsu YH, Venners SA, Terwedow HA, Feng Y, Niu T, Li Z, Laird N, Brain JD, Cummings SR, Bouxsein ML, Rosen CJ, Xu X (2006) Relation of body composition, fat mass, and serum lipids to osteoporotic fractures and bone mineral density in Chinese men and women. The American journal of clinical nutrition 83:146-154. 2. Zhao LJ, Liu YJ, Liu PY, Hamilton J, Recker RR, Deng HW (2007) Relationship of obesity with osteoporosis. The Journal of clinical endocrinology and metabolism 92:1640-1646.[br][br]Sources of Research Support: National Medical Research Council of Singapore.[br][br]Nothing to Disclose: GW-KN, AY-YL, LLC, KV, EST, AC-MN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1199 60 441 SAT-351 PO05-01 Saturday 380 2012


381 ENDO12L_SAT-352 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Loss of Bone Strength in Response to Exercise-Induced Weight Loss in Obese Postmenopausal Women Karen L Villalon, Vanessa D Sherk, Tracy A Swibas, Wendy M Kohrt University of Colorado Denver Anschutz Medical Campus, Aurora, CO Background: Both diet- and exercise-induced weight loss result in loss of areal bone mineral density (aBMD)(1-5), even when fat-free mass was preserved (3-6). It is unknown whether this loss of aBMD translates into decreased bone strength. The current gold standard for assessing aBMD, dual-energy X-ray absorptiometry (DXA), does not distinguish between changes in cortical and trabecular bone. This limits the ability to detect changes in key determinants of bone strength and fracture resistance due to weight loss (7). Quantitative computed tomography (QCT) measures total, cortical, and trabecular volumetric BMD (vBMD) and bone strength at clinically important sites. Herein, we report changes in aBMD, vBMD, and bone strength after exercise-induced weight loss. Methods: Subjects were healthy, postmenopausal women (n=13) who participated in a 4-mo supervised endurance exercise-induced weight loss (WL) intervention. A weight stable (WS) control group (n=8) was also followed for 4-mo. DXA and QCT scans were performed at baseline (BL) and 4-mo to assess changes in hip aBMD and vBMD. Bone strength measures (cross-sectional moment of inertia (CSMI) and section modulus (Z) were obtained for the hip using commercial QCT software (Mindways QCT Pro). Data are reported as mean[plusmn]SD. Results: At study entry, WL women were aged 54.8[plusmn]2.4 y, BMI was 31.6[plusmn]5.8 kg/m2, and total hip t score 0.1[plusmn]1.3. Participants lost -3.0+2.7 kg. WS women were aged 54.4[plusmn]3.3y, BMI was 28.2[plusmn]4.0 kg/m2, and total hip t score was 0.6[plusmn]0.5 (no significant group differences). There were significant decreases in CSMI (p[lt]0.0001) and Z (p=0.002) in the WL group, adjusted for BL values. However, total hip aBMD or vBMD did not change significantly in response to WL. WS women had a significant decline in CSMI (p=0.0002), but not Z, aBMD or vBMD. Conclusions: Given the small subject number, limited amount of weight loss, and relatively short intervention period, no significant decreases in total hip aBMD or vBMD were appreciated. However, there were significant decreases in bone strength measures in response to weight loss. This suggests that changes in bone strength may be more apparent than changes in BMD with weight loss. Further studies are needed to elucidate the mechanisms involved in the loss of bone strength and determine whether such decreases lead to increased fracture risk.[br][br](1) Villareal DT, Chode S, Nehu P, Sinacore DR, Hilton T, Armento-Villareal A, et al. Weight Loss, Exercise, or Both and Physical Function in Obese Older Adults. N Engl J Med 2011; 364(13): 1218-29. (2) Gozansky WS, Van Pelt RE, Jankowski CM, Schwartz RS, Kohrt WM. Protection of bone mass by estrogens and raloxifene during exercise-induced weight Loss. J Clin Endocrinol Metab 2005 Jan; 90(1):52-9. (3)Park HA, Lee JS, Kuller LH, Cauley JA. Effects of weight control during the menopausal transition on bone mineral density. J Clin Endocrinol Metab 2007; 92(10):3809-15. (4) Villareal DT, Fontana L, Weiss EP, Racette SB, Steger-May K, Schechtman KB, et al. Bone mineral density response to caloric restriction-induced weight loss or exercise-induced weight loss: a randomized controlled trial. Arch Intern Med 2006; 166(22):2502-10. (5) Villalon KL, Gozansky WS, Van Pelt RE, Wolfe, P, Jankowski, Schwartz RS, et al. A Losing Battle: Weight Regain Does Not Restore Weight Loss-Induced Bone Loss in Postmenopausal Women. Obesity 2011; doi : 10.1038/oby.2011.263. (6) Lang TF, Li J, Harris ST, Genant HK. Assessment of vertebral bone mineral density using volumetric quantitative CT. J Comput Assist Tomogr 1999; 23(1):130-7.[br][br]Sources of Research Support: NIA F32 AG035460-02.[br][br]Nothing to Disclose: KLV, VDS, TAS, WMK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2000 60 442 SAT-352 PO05-01 Saturday 381 2012


382 ENDO12L_SAT-353 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Increased Bone Turnover in Gastric Bypass Is Corrected after Feeding: A Bone Protector Mechanism in Bariatric Surgery? Juan Patricio Valderas, Sandra Solari, Oslando Padilla, Jeannette Gutierrez, Gilberto B Gonzalez School of Medicine, Pontificia Universidad Cat[oacute]lica de Chile, Santiago, Chile; School of Medicine, Pontificia Universidad Cat[oacute]lica de Chile, Santiago, Chile; School of Medicine, Pontificia Universidad Cat[oacute]lica de Chile, Santiago, Chile; School of Medicine, Pontificia Universidad Cat[oacute]lica de Chile, Santiago, Chile We have described that gastric bypass (GB) is associated even in the long-term to an increase in bone resorption, measured by fasting carboxitelopeptide (CTX). On the other hand, in healthy subjects, feeding results in an acute reduction of bone resorption. This response seems to be regulated by glucagon-like peptide 2 (GLP-2), which increases its secretion after GB. Thus, we hypothesized that the acute reduction of bone resorption after feeding is preserved and even increased in patients with GB.[br]Methods: A cross-sectional study was conducted in 15 postmenopausal women (mean[plusmn]SD: 61.8 [plusmn] 4.0 y.o.; BMI: 29.6 [plusmn] 3.9 kg/m[sup]2[/sup]) with GB performed 7.9 years before (range 36 to 196 months), matched by age (60.3 [plusmn] 3.6 y.o.) and BMI (29.6[plusmn]4.1 kg/m[sup]2[/sup]) with 15 nonoperated women. All volunteers had not any clinical condition that affect bone metabolism. Plasma PTH, calcium corrected by albumin (Ca), phosphate (P), CTX and GLP-2 were measured in fasting and after ingestion of a standard test meal (STM; 355 kcal, 13 g protein, 50 g carbohydrate and 11 g fat) during 180 minutes (m). Data were analyzed by parametric and non parametric paired sample tests, and generalized estimating equation model for repeated measures.[br]Results: In fasting Ca, P, and PTH were normal and similar in both groups (p[gt]0.05); CTX was higher in GB than controls (0.589[plusmn]0.11 vs. 0.382[plusmn]0.11 ng/ml; p=0.001). STM did not induce changes in Ca or P; PTH decreased significantly at 15, 30 and 45 m in both groups with no differences between them. CTX had a significant decrease after STM in both groups since 30 to 180 m. The percentages of reduction from baseline values were significantly higher in GB compared to controls at 45, 60 and 90 m, reaching a nadir of 42.2% at 90 m in GB vs. 53.9 % at 120 m in controls (p[lt]0.05). There were no differences between GB and controls in GLP-2 in fasting (11.7[plusmn]7.0 vs. 8.5[plusmn]6.2 pmol/l, p=0.2). STM induced increase of GLP-2 levels in both groups, but with values significantly higher in GB vs. controls since 30 to 180m (peak at 30 m: 1,868[plusmn]2,542 vs. 33[plusmn]65 pmol/l; p=0.001).[br]Conclusions: To the best of our knowledge, this is the first study to show that the acute reduction of bone resorption after feeding is preserved in GB. This reduction is higher in GB than controls and it is associated to increasing levels of GLP 2. These findings suggest a new bone protector mechanism in GB that may counteract the increased bone resorption described in fasting in these patients.[br][br]1.- Valderas JP et al., Obes Surg 2009; 19:1132. 2.- Walsh JS, Henriksen DB., Arch Biochem Biophys. 2010;503:11.[br][br]Sources of Research Support: This study was funded by a grant from Sociedad Chilena de Endocrinolog[iacute]a y Diabetes (Project N[deg] 2010-03).[br][br]Nothing to Disclose: JPV, SS, OP, JG, GBG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 623 60 443 SAT-353 PO05-01 Saturday 382 2012


383 ENDO12L_SAT-354 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Carbohydrate Beverage Attenuates Bone Resorption Marker in Elite Runners Maysa Vieira Sousa, Rosa Maria Rodrigues Pereira, Maria Elizabeth Rossi Silva University of Sao Paulo, S[atilde]o Paulo, Brazil; University of Sao Paulo, S[atilde]o Paulo, Brazil Runners expose their lower extremity bones and joints to large repetitive axial loads. Such extremes of chronic loading might be expected to affect the turnover rate of bone and cartilage in affected parts of the skeleton and can be detectable using biomarkers assays. Diet and nutrition status have traditionally been the most relevant factors in the management of skeletal health and consideration of this factor has helped to mitigate some losses. Based on that, the aim of this study was to evaluate the effect of carbohydrate beverage on biochemical markers of bone turnover and norepinephrine in long-distance runners undergoing an overload training program. Twenty-four male runners were randomly assigned into two groups: carbohydrates (CHO) and control (CON) groups. The participants were submitted to an overload training program (days 1 - 8), followed by a high-intensity intermittent running protocol (10 x 800 m) on day 9. The runners received maltodextrin solution (CHO group) or zero energy placebo solution as the CON equivalent before, during, and after this protocol. After 8 days of intensive training, osteocalcin baseline levels decreased in both CHO (28.8 [plusmn] 3.6 [italic]vs.[/italic] 24.8 [plusmn] 3.0 ng/ml) and CON group (26.6 [plusmn] 2.4 [italic]vs.[/italic] 21.9 [plusmn] 1.6 ng/ml; p [lt] 0.01). On day 9, immediately after intermittent running (10x800m), the increase in norepinephrine concentrations was lower in the CHO group (1648.0 [plusmn] 204.0 pg/ml) [italic]vs.[/italic] CON group (2542.0 [plusmn] 505.3 pg/ml; p [lt] 0.05) and leptin levels were also lower in the CHO group (166.5 [plusmn] 28.4 pg/ml) [italic]vs.[/italic] CON group (334.1 [plusmn] 66.0 pg/ml; p [lt] 0.05) whereas osteocalcin and P1NP concentrations increased in both groups (p [lt] 0.001). Norepinephrine was positively correlated with P1NP (r = 0.4 and p [lt] 0.001). At 80 min of the recovery period, there was an increase in CTX levels in the CON group (from: 0.4 [plusmn] 0.0 ng/ml to: 0.6 [plusmn] 0.0 ng/ml; p [lt] 0.001) that was diminished with CHO supplementation (from: 0.5 [plusmn] 0.1 ng/ml to: 0.3 [plusmn] 0.1 ng/ml; p [lt] 0.001). Our results show that ingestion of a carbohydrate beverage may result in less bone resorption, providing better recovery during intensive training.[br][br](1) Scott JPR et al., J Clin Endocrinol Metab 2010; 95: 3918-25. (2) Banfi G et al., Sports Med 2010; 40: 697-14.[br][br]Sources of Research Support: State of S[atilde]o Paulo Research Foundation (FAPESP). FAPESP Grant 2007/ 08747-7.[br][br]Nothing to Disclose: MVS, RMRP, MERS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 771 60 444 SAT-354 PO05-01 Saturday 383 2012


384 ENDO12L_SAT-355 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) African American and Caucasian Patients Have Different Risk Factors for Prevalent Vertebral Fractures Sharon H Chou, Maureen Costello, Tamara J Vokes University of Chicago, Chicago, IL There is controversy regarding racial differences in the prevalence of vertebral fractures. While the Study of Osteoporotic Fractures reported a prevalence of 10.6% in African American (AA) and 19.1% in Caucasian (CA) women (1), we found that in women referred for bone mineral density (BMD) testing the prevalence of vertebral fractures was 21% for both races (2). The current study aimed to assess the prevalence of and risk factors for radiographic vertebral fractures detected using VFA (Vertebral Fracture Assessment, spine image obtained on a densitometer) in an independent sample of densitometry patients. Women over 50 years and patients using glucocorticoids (at least 5 mg of prednisone or equivalent for at least 3 months) were invited to participate. Among 336 participants (52 men), there were 183 AA and 151 CA with the mean age ([plusmn]SD) of 69.8 [plusmn] 12.6 and 64.8 [plusmn] 11.7, respectively (p=0.002). Vertebral fractures were graded according to Genant[apos]s semi quantitative method with grade [ge]2 classified as fractured. There was no significant difference in the prevalence of vertebral fractures between AA (16%) and CA patients (18%, p=0.846). In univariate logistic regression analysis, significant risk factors for presence of vertebral fracture were age (OR 1.6, CI 1.3-2.6/decade, p [lt]0.001), height loss (OR 1.3, CI 1.1-1.6/inch, p=0.004), and hip BMD (OR 1.8, CI 1.3-2.3/unit decrease in T-score, p [lt]0.001). In multivariate analysis, the only significant risk factors were age in AA (OR 1.9, CI 1.2-3.2/decade, p=0.008) and hip BMD in CA subjects (OR 2.8, CI 1.7-4.7/unit decrease in T-score, p[lt]0.001). Corticosteroid use and history of non-vertebral fractures were not significant risk factors, possibly due to small sample size. Results were similar when men were excluded from analysis. We conclude that in contrast to population studies, in patients referred for bone densitometry, prevalence of vertebral fractures is similar in AA and CA patients. Our results also suggest that the risk factors for vertebral fractures may differ between the two races with BMD being more predictive in CA and age in AA subjects.[br][br](1)Cauley et al., J Bone Miner Res 2008;23:1458. (2)Vokes et al., J Clin Densitom 2007;10:1.[br][br]Nothing to Disclose: SHC, MC, TJV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1321 60 445 SAT-355 PO05-01 Saturday 384 2012


385 ENDO12L_SAT-356 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Visual Radiographic Texture Analysis of Calcaneal Images in Assessing Bone Fragility Jessica Lanin Hwang, Tamara Vokes University of Chicago Medical Center, Chicago, IL Background: Bone fragility is determined by bone quantity measured as bone mineral density (BMD) and by bone quality or structure that can not be easily assessed using currently available methods. We have previously shown that Radiographic Texture Analysis (RTA, computerized analysis of the pattern of bone radiographs) of calcaneal images obtained on a densitometer can be used to assess bone fragility. We reported that RTA differentiated subjects with and without high bone fragility (defined by presence or absence of vertebral fractures, VFx)1. However, RTA requires image processing and specialized software that may not be readily available in clinical practice. In this study we examined whether visual assessment with semi-quantitative scoring of geometric patterns and contrasts captures qualities similar to RTA. This method, which we termed Visual Radiographic Texture Analysis (VRTA) is potentially more efficient, accessible and inexpensive. In this study we examined whether VRTA correlated with previously obtained computerized RTA measurements and how well VRTA differentiates subjects with and without prevalent VFx.[br]Methods: In 690 women referred to the University of Chicago densitometry unit, BMD was measured at the lumbar spine, proximal femur and calcaneus. RTA and VRTA were performed on densitometric heel images. Fragility was defined as presence of VFx detected on VFA (Vertebral Fracture Assessment - densitometric spine image). The VRTA features included trabecular prominence (PROM) and trabecular length (LENGTH), each scored between 1 and 3 with the higher number corresponding to a more pronounced trabecular pattern.[br]Results: There was a modest but significant correlation between RTA and both PROM (R2= 0.57, p[lt]0.001) and LENGTH (R2=0.50, p[lt]0.001). When controlling for BMD (the lowest T-score from the lumbar spine or proximal femur), a 1 unit decrease in either PROM or LENGTH was associated with the presence of VFx with an odds ratio of 1.67 (95% CI: 1.17-2.37, p=0.004) or 1.65 (95% CI: 1.17-2.31, p=0.004), respectively. The odds ratios for the association of VFx with BMD, expressed per 1 unit decrease in T-score were 1.38 (95% CI: 1.19-1.59, p[lt]0.001) for spine, 2.01 (95% CI: 1.63-2.48, p[lt]0.001) for femoral neck, and 1.39 (95% CI: 1.19-1.61, p[lt]0.001) for heel.[br]Conclusion: VRTA may be useful as a simple and inexpensive way of assessing bone structure and fragility.[br][br](1) Vokes et al: Radiographic Texture Analysis of Densitometric Calcaneal Images - Relationship to Clinical Characteristics and to Bone Fragility. 2010 J Bone Miner Res. 25:56-63.[br][br]Nothing to Disclose: JLH, TV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 894 60 446 SAT-356 PO05-01 Saturday 385 2012


386 ENDO12L_SAT-357 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Do Nutritional Factors Play a Role in the Higher Prevalence of Osteoporosis on Spine in Korean Adults Aged 50 Years or Older? Yong Jun Choi, Bu Kyung Kim, Yoon-Sok Chung Ajou University School of Medicine, Suwon, Republic of Korea Introduction: It has been reported that the vertebral fracture rate is higher in Asians than Caucasians. It is well known that Asians and Caucasians have different life styles, especially in diet. In this study, we calculated the proportion of older adults who have osteoporosis at each skeletal site and investigated if nutritional factors affect osteoporosis at the lumbar spine[br]Methods: We used data from the Fourth Korea Health and Nutrition Examination Surveys (KNHANES) to examine the prevalence of osteoporosis according to the skeletal sites in adults aged [ge]50 years. We divided the subjects with osteoporosis into three groups, spine (osteoporosis only at the spine), femur (osteoporosis only at the femur), and spine and femur osteoporosis (at both spine and femur) groups. The 24-h dietary recall method was used to collect data on food items consumed by participants during the previous 24 h. Information on supplement use was also collected. We compared nutritional factors between the spine and femur groups.[br]Results:In men, 4.2% had osteoporosis only at the lumbar spine, 1.5% only at the femur and 1.8% at both the spine and femur. In women, 13.1% had osteoporosis only at the lumbar spine, 8.4% only at the femur and 14% at both the spine and femur. The mean age of the spine osteoporosis group was younger than the femur group. After adjustment for age and weight, only energy and carbohydrate intake were significantly higher in only the female spine osteoporosis group only in women (p=0.006 and p=0.001, respectively). There were no significant differences in protein or fat intake between the two groups in men and women. There was also no significant difference in vitamin D level between two groups. Energy intake was significantly associated with only the femur neck (P[lt]0.001: men; p=0.035: women) and total hip (p[lt]0.001: men; p=0.019: women) bone mineral density (BMD), but not lumbar spine BMD in both sexes.[br]Conclusion: More Korean older adults had osteoporosis only at the lumbar spine, compared to other races. Nutritional factors were not significantly associated with lumbar BMD. Genetic factors may play a more important role in the ethnic difference in vertebral fracture rather than the nutritional factors.[br][br]Nothing to Disclose: YJC, BKK, Y-SC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 511 60 447 SAT-357 PO05-01 Saturday 386 2012


387 ENDO12L_SAT-358 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Mild to Moderate Renal Dysfunction Is Not a Risk Factor for Low Bone Mineral Density in Korean Men and Women Se Hwa Kim, Soo-Kyung Kim, Tae Ho Kim, Jae Hyuk Lee Kwandong University College of Medicine, Goyang, Korea; CHA University School of Medicine, Seongnam, Korea [bold]Background[/bold]: The objective of this study was to examine whether renal dysfunction was associated with low bone mineral density (BMD) in Korean men and women.[br][bold]Subjects and Methods[/bold]: We analyzed data on 1,602 postmenopausal women and men aged 50 and more from the Fourth Korea National Health and Nutrition Examination Surveys (NHANES IV, 2008). Creatinine clearance (CCr) was calculated using the Cockcroft-Gault (CG) equation. BMD at lumbar spine and proximal femur was measured by dual energy x-ray absorptiometry. Renal function was categorized by the criteria of the Kidney Disease Outcomes Quality Initiative (K/DOQI) chronic kidney disease (CKD) stage.[br][bold]Results[/bold]: The average age was 63.7 [plusmn] 9.0 years. Based on creatinine clearance, 13.9% of subjects were CKD stage 1, 51.8% were CKD stage 2, and 34.3% were staged 3 or 4. There was a positive correlation between CCr and femoral neck BMD (r = 0.414, [italic]p[/italic] [lt] 0.001 in men, r = 0.485, [italic]p[/italic] [lt] 0.001 in women). This association was strongest in those with CKD stage 3 or greater. However, multiple regression analysis showed the association between CCr and femoral neck BMD was extinguished after adjusting for age and weight.[br][bold]Conclusion[/bold]: Mild to moderate renal dysfunction was common in Korean men and women. Although renal function measured by CG was associated with BMD at lumbar spine and femoral neck, this association can be explained by confounding factors such as age and weight.[br][br]Nothing to Disclose: SHK, S-KK, THK, JHL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 395 60 448 SAT-358 PO05-01 Saturday 387 2012


388 ENDO12L_SAT-359 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Relation of Vitamin D Levels with Bone Mineral Density and Parathyroid Hormone in Adults with Low Bone Density Sunil K Kota, Siva K Kota, Svs Krishna, Lalit K Meher, Kirtikumar D Modi Medwin Hospital, Hyderabad, India; Central Security Hospital, Riyadh, Saudi Arabia; MKCG Medical College, Berhampur, India Objective:To investigate relationship among serum 25-hydroxyvitamin D [25(OH)D] levels,parathyroid hormone(iPTH) levels and bone mineral density(BMD).[br]Methods:Adults with low BMD at the total hip or lumbar spine underwent assessment.Multivariate regression models were used to investigate the relationships among serum 25(OH)D,iPTH and BMD.[br]Results:102 patients(M:F=38:64) with mean age of 62.5[plusmn]6.4 years were inducted into the study. Osteoporosis and osteopenia were present in 44 and 58 patients respectively.The mean serum 25(OH)D and IPTH levels were 21.3[plusmn]0.5 ng/ml and 53.1[plusmn]22.3 pg/ml respectively.In 84.3% of patients,serum 25(OH)D levels were below 30 ng/mL. There was no association between 25(OH)D levels and BMD at the total hip, or lumbar spine (P=.473, and.353 respectively).Serum iPTH levels were negatively associated with BMD at the total hip (P=.019) and the lumbar spine (P=.02).Both at the total hip and lumbar spine,iPTH levels,male sex,BMI and age were found to be significant predictors of BMD.Patients with higher BMI had significantly lower BMD and T score.At levels [lt]30 ng/mL,25(OH)D was negatively associated with iPTH (P=.041).[br]Discussion: Vitamin D deficiency is a risk factor for osteoporosis.Some studies have reported a positive correlation between 25(OH)D levels and BMD at all sites(1),others finding a correlation at femoral neck but not at other sites(2).In osteoporotic patients,a positive correlation between 25(OH)D levels and BMD at the femoral neck but not at the lumbar spine was found(3); another study found an association between serum 25(OH)D and BMD at the trochanter only in patients with a serum 25(OH)D concentration [lt]10 ng/mL (4).A south east Asia study including patients with low BMD revealed no association between 25(OH)D levels and BMD at the femoral neck,total hip or lumbar spine(5).[br]Conclusion: Among our cohort of patients with low BMD, no direct relationship between serum 25(OH)D levels and BMD at total hip and lumbar spine was observed.A negative correlation existed between iPTH and 25(OH)D at serum 25(OH)D concentrations [lt]30 ng/mL and serum iPTH levels showed a significant negative association with BMD at the total hip and lumbar spine.These significant negative associations between iPTH levels and BMD at the total hip and lumbar spine underscore the critical role of this hormone in bone metabolism and health.Advancing age,male sex,BMI are other signifcnat predictors for BMD both at total hip and lumbar spine.[br][br](1) Roy DK, et al., Bone. 2007;40:200. (2) Ooms ME, et al.,J Bone Miner Res. 1995;10:1177. (3) Aguado P, et al., Osteoporos Int. 2000;11:739. (4) Lips P, et al., J Clin Endocrinol Metab. 2001;86: 1212. (5) Chandran M, et al., Endocr Pract. 2011;17:226.[br][br]Nothing to Disclose: SKK, SKK, SK, LKM, KDM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1344 60 449 SAT-359 PO05-01 Saturday 388 2012


389 ENDO12L_SAT-360 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Gender Difference in Osteoporosis Screening: Retrospective Chart Review Khaled Alswat, Suzanne Myers Adler George Washington University, Washington DC Veterans Affairs Medical Center, Washington, DC; George Washington University, Washington, DC [bold][underline]Background:[/underline] [/bold]Osteoporosis is a common disease affecting [sim]30% of all postmenopausal women. It accounts for [gt]1.5million fractures yearly in the USA. Up to 20% of patients who sustain hip fractures die within the first 12 months from related complications.[sup](1) [/sup]Physician-specific osteoporosis screening rates of a random sample of women aged [gt]65 years vary widely, ranging from 19-97%. Overall, the mean rate of osteoporosis screening among all physicians is 56 %.[sup](2) [/sup]There are little data comparing gender-specific osteoporosis screening rates. The National Osteoporosis Foundation recommends screening all women [gt]65 and men [gt]70 years of age regardless of risk factors. The aim of the study is to identify any gender difference in osteoporosis screening.[br][bold][underline]Methods:[/underline] [/bold]We conducted a retrospective study to determine the screening rates for osteoporosis in males and females in our Division of Internal Medicine, university based outpatient clinic (UBC). Males between the ages of 70-75 years and females between the ages of 65-70 years with a primary care physician (PCP) at our UBC, who have had at least one routine health maintenance exam (HME) since 2002 were included. We counted the number of patients on whom DEXA was performed in order to determine DEXA screening rates for both males and females. We excluded patients who have had DEXA at our facility but without a PCP in our Division of Internal Medicine; however, these patients were included to calculate the overall rate of osteoporosis.[br][bold][underline]Result:[/underline] [/bold]A total of 8,262 patients who met the age criteria were identified; 3,255 (39.4%) males and 5,007 (60.6%) females.[br]Of the 3,255 male patients, 1,415 (43.5%) had their PCP outside our UBC and were excluded, and 1,840 (56.5%) males had their PCP at our UBC and were included. Of this group, 342 had a HME and 63 had DEXA performed for an osteoporosis screening rate of 18.4%.[br]Of the 5,007 female patients, 2,354 (47%) had their PCP outside our UBC and were excluded, and 2,653 (53%) had their PCP at our UBC and were included. Of this group, 668 patients had a HME and 402 had DEXA performed for an osteoporosis screening rate of 60%.[br]18.2% of males and 19.3% of females who had DEXA performed had findings consistent with osteoporosis.[br][bold][underline]Conclusion:[/underline] [/bold]The female screening rate at our UBC for those who had a HME visit is similar to the overall mean rate for osteoporosis screening reported in other studies.[sup](2) [/sup]Males are screened less frequently although they have a comparable prevalence of osteoporosis.[br][br](1) Johnell O,Kanis JA. An estimate of the worldwide prevalence and disability associated with osteoporotic fractures. Osteoporos Int. 2006 Dec; 17(12):1726-33. (2) Kenneth Cohen. Evaluation of Screening Patterns in a Primary-Care Group Practice. Journal of clinical densitometry. Pages 498-502 (October 2008).[br][br]Nothing to Disclose: KA, SMA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1176 60 450 SAT-360 PO05-01 Saturday 389 2012


390 ENDO12L_SAT-361 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Elevated Serum Homocysteine Levels Are Associated with Vertebral Fracture in Type 2 Diabetic Men Masahiro Yamamoto, Toru Yamaguchi, Toshitsugu Sugimoto Shimane University Faculty of Medicine, Izumo, Japan Introduction: We previously reported that patients with type 2 diabetes (T2DM) had an increased risk of vertebral fractures (VFs) despite higher bone mineral density (BMD) compared to non-T2DM controls (1), and that pentosidine, which is one of well-known advanced glycation end-products, and its endogenous secretory receptor were associated with VFs independent of BMD (2, 3), suggesting that pentosidine is attributed to the pathogenesis of poor bone quality in T2DM patients. Previous reports show that urinary pentosidine levels are correlated with serum homocystein (HC) levels in postmenopausal women and that elevated HC levels are associated with an increased risk for hip fracture in older men and women. However, it is still unclear whether HC levels are associated with VFs in T2DM patients. In this study, a relationship between serum levels of HC and presence of VFs was examined in T2DM patients.[br]Methods: Two hundred-two men older than 50 yr and 140 postmenopausal women with T2DM within normal creatinine levels were examined by bone mineral density (BMD) at the lumbar spine, and femoral neck (FN) as well as spine radiographs, and by measurement of biochemical parameters including serum HC, and pentosidine.[br]Results: Simple regression analysis revealed that age, BMI, HbA1c, pentosidine and BMD except for female FN were not associated with HC levels in either gender. Serum creatinine levels in both genders as well as female FN-BMD were positively and inversely correlated with HC levels, respectively, and these relationship still remained significant after adjustment for age, BMI, HbA1c and duration of T2DM (creatinine: Men, r = 0.29, P [lt] 0.01, Women, r = 0.23, P = 0.01; FN-BMD, r = -0.27, P [lt] 0.01). HC levels in men with VFs were significantly higher than those without VFs. Multivariate logistic regression analysis adjusted for age, BMI, HbA1c, creatinine, duration of T2DM, and FN-BMD identified HC level as an factor associated with the VFs in T2DM men [odds ratio 2.41, (95% confidence interval 1.21[ndash]4.79), P = 0.01 for each increase of 1SD in HC levels].[br]Conclusion: The present findings showed that serum HC level were associated with prevalent VFs independent of BMD in T2DM men, suggesting that HC may be related to bone quality that is not defined by BMD. Thus, measurements of serum HC levels seem to be helpful for assessing VF risk in patients with T2DM men.[br][br](1)Yamamoto M et al. J Bone Miner Res 2009; 24:702. (2)Yamamoto M et al. J Clin Endocrinol Metab 2008; 93:1013. (3)Yamamoto M et al. Diabetes Care 2009; 32:2263.[br][br]Nothing to Disclose: MY, TY, TS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1043 60 451 SAT-361 PO05-01 Saturday 390 2012


391 ENDO12L_SAT-362 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Osteoporosis in Type 2 Diabetes Mellitus: No Sex Bias [amp] Both Postmenopausal Diabetic and Non-Diabetic Patients Are Equally Affected Kalpana Dash, Chiranjit Roy Apollo Hospitals, Bilaspur, India; Apollo Hospitals, Bilaspur, India [bold]Objective:[/bold] To assess the association of diabetes and osteoporosis/osteopenia in men and women and association with the severity of diabetes.[br][bold]Methods:[/bold] We screened 100 patients aged 30-75 years of age coming to Endocrine clinic from September 2009 and January 2010. All of them were subjected to undergo BMD scan. HbA1c was done in all patients by Turbidometry method. Bone mineral density measurement of hip area and the lumbar spine were done by using the Dual Energy X-ray Absorptiometry scan. Osteoporosis and osteopenia were assessed on the basis of the World Health Organization guidelines.[br][bold]Results:[/bold] There were 36 male and 64 female patients with an average age of 55.6 years and 50.89 years respectively. There were 52 diabetic (M-30, F-22) and 48 (M-7, F-41) non diabetic individuals. Bone mineral density analysis revealed osteoporosis/osteopenia in 87 [M- 30 (34.48%), F-57 (65.52%)] patients. Among osteoporosis/osteopenia patients, 47(54.02%) were diabetic [amp] 40 (45.97%) were non diabetic patients. 24 (51.06%) male and 23 (48.93%) female diabetic individuals had osteoporosis/osteopenia. Among non diabetic individuals, 6 (15%) men and 34(85%) women had osteoporosis/osteopenia. Among 64 female individuals 36 (56.6%) were post menopausal and 28 (45.4%) were premenopausal. Among post menopausal patients 36 (58.3%) were diabetics [amp] 15 (41.6%) were nondiabetics. Interestingly, both post menopausal diabetics and nondiabetics individuals were equally affected by osteoporosis and osteopenia 20(95.2%) and 14 (93.3%) respectively. Hence, this retrospective analysis shows, there was no difference in the prevalence of osteoporosis/osteopenia in diabetic and non-diabetic post menopausal women and that both men and women diabetics were equally affected by osteoporosis/osteopenia. The average HbA1c value was 7.9%, when correlated with BMD measurement was found that, higher the HbA1c level, higher was the severity of osteoporosis/osteopenia.[br][bold]Conclusion:[/bold] We conclude that, the prevalence of osteoporosis/osteopenia was equal among diabetic men and women (M-51%, W-49%) where as in non diabetic individuals, women 34 (85%) were more affected than men 6 (15%). Hence there is no sex bias in diabetic patients having osteoporosis/osteopenia [amp] this could be probably due to glycemic control modifying the pathophysiology of osteoporosis/osteopenia. Further, there is no difference in prevalence of osteoporosis/osteopenia among post menopausal diabetic and nondiabetics patients.[br][br]1.Sta Romana M, Li-Yu JT.Investigation of the relationship between type 2 diabetes and osteoporosis using Bayesian inference,2007 Oct-Dec;10(4):386-90. 2.Liu EY, Wactawski-Wende J, Donahue RP, Dmochowski J, Hovey KM, Quattrin T. Does low bone mineral density start in post-teenage years in women with type 1 diabetes? Diabetes Care. 2003;26:2365-2369. 3.Rosato MT, Schneider SH, Shapses SA. Bone turnover and insulin-like growth factor 1 levels increase after improved glycemic control in noninsulin-dependent diabetes mellitus. Califc Tissue Int. 1998; 63:107-111. 4.Schwartz AV, Sellmeyer DE, Ensrud KE, et al. Study of osteoporotic features research group. Older women with diabetes have an increased risk of fracture: a prospective study. J Clin Endocrinol Metab. 2001;86:32-38. Abstract.[br][br]Sources of Research Support: Self.[br][br]Nothing to Disclose: KD, CR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1484 60 452 SAT-362 PO05-01 Saturday 391 2012


392 ENDO12L_SAT-363 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Serum Sclerostin Levels in Men with Idiopathic Osteoporosis Bruno Lapauw, Sara Vandewalle, Youri Taes, Stefan Goemaere, Hans-Georg Zmierczak, Julien Collette, Jean-Marc Kaufman Ghent University Hospital, Ghent, Belgium; Ghent University Hosptial, Ghent, Belgium; CHU Sart Tilman, Li[egrave]ge, Belgium BACKGROUND: Our previous findings in a family-based study of men with idiopathic osteoporosis concurred to indicate the existence of a familial defect in the acquisition of peak bone mass and size. Sclerostin, an osteocyte-specific glycoprotein, is a strong negative regulator of osteoblast activity through the Wnt/[beta]-catenin-pathway. If sclerostin is involved in the less efficient bone acquisition in men with idiopathic low bone mass, this might be reflected in higher serum sclerostin levels.[br]OBJECTIVE: To assess serum sclerostin levels and relate them to bone parameters in men with idiopathic low bone mass.[br]DESIGN: Cross-sectional study of 114 men diagnosed with idiopathic low bone mass ([lt] 65 yrs), compared to 114 healthy age-matched control men.[br]METHODS: Areal bone mineral content (BMC) at the spine, proximal femur and whole body measured using dual X-ray absorptiometry (Hologic). Volumetric and geometric bone parameters at the radius (4% and 33% site) assessed using peripheral quantitative computed tomography (pQCT - XCT2000, Stratec GmbH). Serum sclerostin levels measured using an immunoassay (Biomedica Medizinprodukte GmbH, Vienna).[br]RESULTS: Men with idiopathic osteoporosis had lower levels of sclerostin (0,54[plusmn]0,17 vs.0,66[plusmn]0,23 ng/mL; P[lt]0.001). In probands and healthy controls, sclerostin levels were strongly associated with age (r=0,46; P[lt]0.001); when adjusting for age, no associations with weight or height were observed (P[gt]0,14). When adjusting for age, weight and height, sclerostin levels displayed a positive association with whole body BMC (r=0,21; P=0.014) in the probands. No associations were observed in the control group, neither were sclerostin levels associated with BMC at the radius or lumbar spine (all P[gt]0,11). Except for a positive trend with cortical thickness (r=0,20; P=0.055), no associations were found between sclerostin levels and pQCT-derived trabecular or cortical bone parameters (all P[gt]0,13).[br]CONCLUSIONS: Lower rather than higher serum sclerostin levels in probands with idiopathic osteoporosis suggest that sclerostin is not involved in the pathogenesis of low bone mass in these subjects. The results rather suggest that the lower serum sclerostin levels in men with idiopathic osteoporosis reflect their lower bone mass and osteocyte number.[br][br]Nothing to Disclose: BL, SV, YT, SG, H-GZ, JC, J-MK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1225 60 453 SAT-363 PO05-01 Saturday 392 2012


393 ENDO12L_SAT-364 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Prevalence of Morphometric Vertebral Fracture in Old Men and the Agreement between Different Methods in the City of Recife, Brazil Juliana Coelho Maia, Denise Dantas Lima, Carla Nubia Borges, Marcelo Cabral, Paula Aragao Prazeres, Francisco Alfredo Bandeira University of Pernambuco Medical School, Recife, Brazil Introduction: Osteoporosis is relatively common in men and has a great impact on quality of life, although it[apos]s often neglected by clinicians worldwide. Despite the importance of the subject, there are few studies that provide information about prevalence of morphometric vertebral fracture in men and the associated risk factors. Objective: The aim of the present study was to determine the prevalence of morphometric vertebral fractures in elderly men by three methods, the agreement between the different methods and its association with solar index, skin phototype and metabolic factors. Methods: 234 men aged [gt] 60 years (mean age 69,4 +- 6,5 years) from a primary care district were evaluated using lateral radiograph of the thoracolumbar spine that were analyzed by two experienced radiologists according to two methods: Genant and Jiang. A third senior radiologist adjucated Genant[apos]s method. Results: The highest prevalence of fractures in Jiang and Genant methods were 37.6% and 36.8% respectively (both by examiner 2). The lowest prevalence rates were 26.5% in Jiang method and 5.6% in Genant (both by examiner 1). The prevalence found by the Genant adjucated was 31.6%. The agreement between the examiners were 69.2% in the method of Jiang (Kappa 0.30; 95% CI = 0.17 to 0.43) and 65.5% in the method of Genant (Kappa 0.09; 95% CI = 0.01 to 0.17).[br]The agreement between the methods by examiners showed a greater coincidence in the second examiner (98.3%) with a kappa of 0.96 (95% CI = 0.93 to 1.0). We evaluated skin phototype, waist circumference, the presence of hypertension, body mass index (BMI), history of fracture, calcium intake, serum 25 (OH) vitamin D and solar index. After using multivariate regression analysis we found that lower BMI (prevalence ratio = 1.41; p = 0.024; 95% CI = 1.05 to 2.03) and solar index (prevalence ratio = 1.45; p = 0.049; 95% CI = 1.01 to 1.95) were independently associated with morphometric vertebral fracture. Conclusion: Our results found a high prevalence of morphometric vertebral fractures in elderly men and a high concordance between the examiners in the method of Jiang. The solar index and BMI were inversely associated with morphometric vertebral fractures, suggesting frailty as a common factor.[br][br]Nothing to Disclose: JCM, DDL, CNB, MC, PAP, FAB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1296 60 454 SAT-364 PO05-01 Saturday 393 2012


394 ENDO12L_SAT-365 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Specific Phytoestrogen Intake Is Associated with Higher Bone Mineral Density in Young Men from Southern China Elaine YN Cheung, Ching-Lung Cheung, Shivani Sahni, Kathryn CB Tan, Annie WC Kung United Christian Hospital, Hong Kong, Hong Kong; University of Hong Kong, Hong Kong, Hong Kong; Hebrew Senior Life and Harvard Medical School, Boston, MA Introduction: It has been shown that phytoestrogens may help preserve bone mineral density (BMD) in postmenopausal women. However, such data is relatively limited among young men and premenopausal women. The aim of this study was to examine the cross-sectional associations between phytoestrogens intake (isoflavones, coumestrol, flavonoids and lignans) and BMD at lumbar spine (LS) and femoral neck (FN) in a cohort of Southern Chinese young men and premenopausal women.[br]Methods: 386 men and 957 women with completed food frequency questionnaire and BMD measured by dual X-ray absorptiometry between 1998 and 2009 were included in this study. Linear regression analysis was used to examine the association between different classes of phytoestrogens intake (as continuous variable and sex-specific tertiles) and BMD at LS and FN. All models were adjusted for age, BMI, calcium intake, smoking and drinking history, number of pregnancy (for women), educational and exercise level, serum estradiol (for women and men) or testosterone (for men) level. A linear test for trend was conducted across tertiles of phytoestrogen intakes.[br]Results: For both sexes, the mean age at baseline was 32[plusmn]5years (range 20-39years). In young men, dietary lignan intake was significantly associated with lumbar spine (beta=0.093; p=0.008) and femoral neck (beta=0.071; p=0.041) BMD whereas flavonoid intake was significantly associated with lumbar spine BMD (beta=0.075; p=0.032) but not femoral neck BMD (beta=0.046; p=0.182). Interestingly, isoflavone intake was negatively associated with lumbar spine BMD (beta=-0.024; p=0.011) but not femoral neck in men. In pre-menopausal women, only marginal associations were observed for lignan and flavonoid intake with BMD at both sites (P range: 0.061-0.09) while no significant associations were observed for other classes of phytoestrogen intake and BMD (P range: 0.214-0.96). After adjustment, men in the highest tertile of lignan intake had higher LS BMD (P-trend = 0.02) and FN BMD (P-trend = 0.01) compared to subjects in the lowest tertile of intake. Similar trend was observed for flavonoid intake and LS (P-trend = 0.036) but not FN BMD. No significant trend of LS BMD was seen with tertiles of isoflavone intake.[br]Conclusion: Significant associations were observed for more than one class of phytoestrogen intake and BMD in young men while marginal associations were observed among pre-menopausal women. Further examination by a prospective study will be needed.[br][br]Nothing to Disclose: EYNC, C-LC, SS, KCBT, AWCK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 801 60 455 SAT-365 PO05-01 Saturday 394 2012


395 ENDO12L_SAT-366 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Safety of Intravenous Pamidronate for Bone Hyperresorption in Patients with Chronic Critical Illness (CCI) and Chronic Kidney Disease (CKD) Rifka C Schulman, Chenbo Zhu, James H Godbold, Jeffrey I Mechanick Mount Sinai School of Medicine, New York, NY [bold]Objective:[/bold] Chronic critical illness (CCI) occurs in survivors of acute critical illness remaining ventilator-dependent via tracheostomy. Pamidronate reduces bone hyperresorption in patients with CCI (1), but its use has been limited due to concerns about nephrotoxicity. The purpose of this study is to determine the safety of intravenous (IV) pamidronate on renal function in patients with CCI and CKD.[br][bold]Methods:[/bold] A retrospective case series of patients admitted to The Mount Sinai Hospital Respiratory Care Unit from 2006-2010, identified patients who received IV pamidronate, administered for 24 hour urine N-telopeptide (NTx) [ge] 70 nmol BCE/mmol Cr or serum NTx [gt] 40 nMBCE/L. Pamidronate 30-90mg was infused intravenously over 4 hours. Patients in both groups received calcium carbonate, ergocalciferol and calcitriol. Changes in glomerular filtration rate (GFR), estimated by the MDRD formula, and creatinine were tabulated for 4 CKD groups for those who did (N=115) and did not (N=200) receive pamidronate, respectively: no or mild CKD up to stage 2 (N=83, N=121), stage 3 CKD (N=13, N=28), stage 4 (N=3, N=30), and stage 5 on hemodialysis (N=16, N=21).[br][bold]Results:[/bold] None of the pamidronate groups or doses showed significant reductions in GFR [ge] 25% immediately after, or at 7 or 14 days post-infusion. Median changes in GFR from baseline to 7 days per CKD group were 0% (P[lt]0.0001), -4% (P=0.29), -4% (P=1) and -9% (P=0.02) after pamidronate, and 0%, 0%, 6% and 6%, in the control groups (P[lt]0.05), respectively. Patients with increases in creatinine [ge] 0.5 or [ge] 1.0 mg/dl (for baseline creatinine [lt] or [ge] 1.4 mg/dl, respectively) were 0%, 4.4%, 6.7% and 18.2%, in the pamidronate groups, and 0.8%, 2.4%, 8.8%, and 20.8%, in the control groups, respectively. Differences in creatinine change between the groups did not reach statistical significance (P=0.17, 0.06, 0.39, 0.78, respectively).[br][bold]Discussion:[/bold] IV pamidronate was not associated with significant worsening of renal function, irrespective of CKD stage. Small fluctuations in renal function in this cohort of patients with multiple comorbidities were not noted to be significant in the pamidronate group compared to the general CCI population.[br][bold]Conclusion:[/bold] Pamidronate was not associated with short-term nephrotoxicity in patients with CCI, with or without CKD. It is hoped that these results will remove barriers to more aggressive management of metabolic bone disease in CCI, which may have salutary downstream effects on morbidity and mortality.[br][br](1) Nierman DM, Mechanick JI. Biochemical response to treatment of bone hyperresorption in chronically critically ill patients. Chest 2000;118(3):761-766.[br][br]Disclosures: JIM: Speaker, Abbott Nutrition; Principal Investigator, Select Medical Corporation. Nothing to Disclose: RCS, CZ, JHG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 372 60 456 SAT-366 PO05-01 Saturday 395 2012


396 ENDO12L_SAT-367 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Diagnosing Remote Bisphosphonate-Associated Atypical Femur Fractures Peter McIntyre, Babette Glister Walter Reed National Military Medical Center, Bethesda, MD Background:[br]In the last decade evidence has emerged linking chronic bisphosphonate use to atypical femur fractures. The pathophysiology of these fractures is widely believed to be weakened microarchitecture from inhibited remodeling. Fractures are often heralded by chronic atraumatic thigh pain in patients using bisphosphonates for over 5 years. Radiologic features include cortical thickening, beaking, and horizontal subtrochanteric lucency before emergence of a transverse subtrochanteric fracture. Changes are frequently found in films of the contralateral leg at the time of fracture, or in review of films evaluating thigh pain from before the fracture. To our knowledge there are no reports of using contemporary films to diagnose bisphosphonate-related fractures from persistant changes after healing.[br]Clinical Case:[br]Our patient is a 55 year old female with osteoporosis diagnosed by DEXA in the mid-1990[apos]s while evaluating recurrent metatarsal stress fractures. She had been on oral alendronate for 5 years when she stepped off a curb and spontaneously fractured her bilateral femurs in 2000. Workup for secondary causes was pertinent for known seronegative rheumatoid arthritis and psoriasis, but she had never used prednisone. She was continued on alendronate because the link to atypical fractures was not yet well-recognized. Over the next 11 years she had recurrent atraumatic rib fractures confirmed by bone scan. Bone turnover markers including serial osteocalcin measurements were in the lower third of the reference range. In 2011 she presented to a new provider with new rib pain. Her workup showed a possible thoracic compression fracture, but T-scores were now normal on DEXA scan. Her long-term (17 year) bisphosphonate use was re-evaluated and plain films were obtained of her femurs, which showed findings characteristic of atypical femur fractures.[br]Clinical Lesson:[br]Atypical femur fracture is a feared complication of long-term bisphosphonate use and has sparked a movement to consider periodic [ldquo]drug holidays[rdquo] from these agents. A frequent clinical dilemma involves patients whose DEXA scans show improvement but clinical history suggests ongoing fracture risk. To our knowledge there are no published reports of using plain X-rays of previously healed fractures to demonstrate bisphosphonate-related adverse events, but in our patient this provided a persuasive argument to stop her bisphosphonate.[br][br]Koh, J, et al. [ldquo]Femoral cortical stress lesions in long-term bisphosphonate therapy: a herald of impendingfracture?[rdquo] J Orthop Trauma, 2010;24(2):75-81. Rosenberg, et al. [ldquo]Bisphosphonate-related complete atypical subtrochanteric femoral fractures: diagnostic utility of radiography.[rdquo] AJR Am J Roentgenol, 2011;197(4):954-60. Ahlman, M, et al. [ldquo]Case review: Evolution of bisphosphonate related atypical fracture retrospectively observed with DXA scanning.[rdquo] J Bone Miner Res, 2011;[Epub ahead of print].[br][br]Nothing to Disclose: PM, BG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 560 60 457 SAT-367 PO05-01 Saturday 396 2012


397 ENDO12L_SAT-368 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) An Unusual Subtrochanteric Fracture in a Patient Receiving Denosumab Rodis Paparodis, Bjoern Buehring, Neil Binkley, Ellaine Pelley University of Wisconsin School of Medicine and Public Health, Madison, WI; University of Wisconsin School of Medicine and Public Health, Madison, WI Background: Atypical subtrochanteric femoral fractures (AFF) are an uncommon complication of osteoporosis therapy. These fractures are often heralded by groin/thigh pain. AFF risk appears to increase with prolonged bisphosphonate therapy. Given this, over-suppression of bone turnover has been invoked as a precipitating factor in the development of these fractures.[br]Methods: We report a case of an unusual femoral fracture after treatment with denosumab. Our patient is an 81-year-old woman who presented with CKD, secondary hyperparathyroidism, osteopenia, and elevated fracture risk. She had a remote history of estrogen therapy but no prior bisphosphonate treatment. She was treated with one injection of denosumab and developed severe right groin/thigh pain approximately six months later. There was no recent history of trauma or falls. Femoral radiography and MRI were performed and a fracture was identified. In order to provide more insight into this fracture, we measured serum markers of bone turnover and reviewed her medical history and imaging.[br]Results: Her femoral radiograph was negative, without any cortical thickening. MRI performed the following day revealed a transverse subtrochanteric insufficiency fracture with a medial cortical defect involving approximately 25% of the femoral cortex. There was associated periosteal reaction medially. Serum bone-specific alkaline phosphatase and N-telopeptide were normal at 9.3 (7.0 - 22.4 ug/L) and 9.5 (6.2 - 19.0 nM BCE) respectively. Levels of serum calcium, phosphate, and 25-(OH) vitamin D were normal. The parathyroid hormone level was elevated at 167 (15-65 pg/mL).[br]Conclusion: The subtrochanteric fracture location, concurrent treatment with antiresorptive therapy, and presentation with thigh/groin pain are reminiscent of bisphosphonate-associated AFF. However, in contrast to most reported AFF cases, this patient presented with an unusual subtrochanteric fracture which occurred (1) with the newer antiresorptive agent denosumab, (2) early in the course of antiresorptive therapy, and (3) in a medial location. Bone turnover markers were not low, suggesting that oversuppression of bone turnover was unlikely to be a contributor to this fracture. The etiology of this fracture remains unclear. Metabolic factors such as hyperparathyroidism may have contributed. However, this case illustrates that not all [ldquo]atypical[rdquo] femoral fractures are related to bisphosphonate therapy.[br][br]Disclosures: NB: Consultant, Merck & Co., Amgen, Lilly USA, LLC, Tarsa; Investigator, Merck & Co., Amgen; Researcher, Tarsa. Nothing to Disclose: RP, BB, EP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 613 60 458 SAT-368 PO05-01 Saturday 397 2012


398 ENDO12L_SAT-369 POSTER SESSION: Osteoporosis (1:30 PM-3:30 PM) Multiple Pathological Fractures in a 41-Year-Old Woman [mdash] Case Report Miro Cokolic, Ursa Ksela University Clinical Centre, Maribor, Slovenia Background: Chronic glucocorticoid excess has deleterious effects on bone that can lead to osteoporosis and fractures.[br]Clinical case: In 9/2003 the 41-year female was examined by the orthopaedist because of the back pain stated under the right spatula and body weight gain. Due to insistence of problems together with conservative therapy, several examinations at neurologist, neurosurgeon and orthopaedist, several X-rays were made, CT radiculography, EMG, US of the hip and the abdomen, sternal puncture, mammography, MR of L/S. Laboratory and tumour markers were normal. In 6/2005 scintigraphy was made and it showed the pathological accumulation in the rib, skull and pelvis. X-ray showed the old fracture of the both pubic bones, for what she was hospitalized in the Dep. of Orthopaedics. Bone biopsy was made, histological examination showed callus development after fracture. She started with bisfosfonate therapy. Problems insisted. An abdominal CT-scan showed bilateral adrenal hyperplasia (right 2,5 cm, left 1 cm). Meanwhile she made gynecological examination to exclude malignancy, once again the rib biopsy was made in the Dep. of Surgery because of newly discovered pathological rib fractures. Orthopaedist sent the patient to endocrinologist, she was first examined in 7/2005. In the status the increased weight, cushinguid look and hypertension stood out. Basic lab. results were normal, osteocalcin suppressed (7 ng/l, n 12-41), [beta]-cross laps (0,27 ng/l, n 0,12-0,40. Hormone testing showed increased cortisol at 8 A.M. (778 nmol/l, n 130-640), elevated cortisol at 1 P.M. (905 nmol/l, n 60-430), normal vitamin D (65,2 [mu]mol/l, n 47,7-144), TSH (1,08 mIU/l, n 0,27-4,2), iPTH (59,3 pg/ml, n 15-65) and ACTH (7,93 pmol/l, n[lt]10,2), no suppression after 1 mg dexamethason overnight test (cortisol 914 nmol/l, n[lt]50). DXA showed osteoporosis(T-score -2,88 SD L1-4 and -1,25 SD left hip, -1,88 SD neck). Adrenal scintigraphy with iodomethyl-19-norcholesterol showed the accumulation in the right adrenal gland. In 7/2006 the right sided laparoscopic adrenalectomy in the Dep. of Urology was made. Histological examination showed the adrenal adenoma. Noteworthy, cortisol excretion fell to near normal levels. Since operation there were no further complications.[br]Conclusion: Spontaneous fractures in Cushing syndrome are very common and they should be taken into consideration. According to the typical clinical picture endocrine cause of patient[apos]s problems was discovered lately.[br][br]Nothing to Disclose: MC, UK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1518 60 459 SAT-369 PO05-01 Saturday 398 2012


399 ENDO12L_SAT-370 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Bisphosphonate-Associated Atypical Hip Fracture with Prodromal Symptomatology Mark Oertel, Candice Rose, Leigh M Eck University of Kansas, Kansas City, KS [bold]Introduction[br][/bold]Bisphosphonates (BP) are the gold standard treatment for osteoporosis (OP) to prevent future fractures. BP use may be causally related to atypical femur fractures particularly in those taking BP for [gt] 5 years. The American Society of Bone and Mineral Research (ASBMR) have proposed criteria to define atypical femoral fractures and recommendations for the evaluation of groin pain in those on BP therapy.[br][bold]Clinical Case[/bold][br]A 74 year old female with glucocorticoid (GC) related OP, on chronic proton pump inhibitory (PPI) therapy, treated with teriparatide therapy x 2 years and BP therapy x 5 years had complaint of left groin pain. A hip radiograph was obtained without abnormal findings. Due to continued pain, an MRI was performed with evidence of a stress fracture in the subtrochanteric aspect of the left femur. Within hours of this MRI and prior to results being reviewed, the patient experienced intense pain with non-traumatic external rotation of her left leg. Imaging revealed a left subtrochanteric fracture with cortical thickening, a medial spike and lateral beaking. Cephalomedullary nailing was pursued. The patient was continued on calcium and vitamin D; no additional therapy for OP was offered given prior two year teriparatide exposure. Laboratory obtained at time of fracture revealed an osteocalcin of 4.2 ng/ml (8.4-55.0), serum crosslinks of 15.5 nMBCE/L (6.2-19), and normal vitamin D status.[br][bold]Clinical Lessons [/bold][br]An ASBMR task force created a case definition for atypical fractures with major criteria to include: subtrochanteric location, non-traumatic event, transverse configuration, and non-comminuted with a medial spike [ndash] all present in this subject. Minor criteria present in this case include beaking of the lateral cortex, cortical thickening, and use of BP/GC/PPI therapy as well as prodromal groin pain. Avoidance of a complete fracture may have been achieved in our patient if limited weight bearing status and/or prophylactic pinning had been pursued. There is likely an association between long-term BP use and atypical fractures in a small number of patients with additional risk factors, including GC or PPI use. All clinicians should be aware of the potential association of atypical fractures with BP use; patients should be asked about groin pain. If present, urgent and thorough evaluation is indicated.[br][br]Nothing to Disclose: MO, CR, LME 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 287 61 460 SAT-370 PO46-02 Saturday 399 2012


400 ENDO12L_SAT-371 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Nonuremic Calciphylaxis Precipitated by Teriparatide [rhPTH(1-34)] Therapy in the Setting of Chronic Warfarin and Glucocorticoid Treatment Elias Spanakis, Deborah Sellmeyer Johns Hopkins Hospital, Johns Hopkins University, Baltimore, MD; Johns Hopkins Bayview Hospital, Johns Hopkins University, Baltimore, MD Background: Calciphylaxis is rare in the absence of end stage renal disease. Predisposing factors for nonuremic calciphylaxis include hyperparathyroidism, coagulopathies, connective tissue disease, liver disease, glucocorticoid use, and malignancy. Warfarin may facilitate vascular calcification by reducing the vitamin K[ndash]dependent carboxylation of matrix-Gla protein.[br]Clinical case: An 86-year old Caucasian woman with a history of polymyalgia rheumatica, two spontaneous deep venous thromboses, and vertebral fractures was treated chronically with calcium, vitamin D, prednisone, and warfarin. Two months after initiating teriparatide therapy for osteoporosis, she developed painful erythematous nodular lesions on her calves bilaterally that progressed to necrotic ulcers on the left leg despite intravenous antibiotic therapy. Biopsy of the lesions showed calcification in the media of small blood vessels and subcutaneous fat with fat necrosis, consistent with calciphylaxis.[br]Teriparatide, calcium, and vitamin D were discontinued and wound care was initiated. Laboratory studies one month later demonstrated a serum creatinine of 1.1 (0.6-1.3 mg/dl), eGFR of 46 ml/min/1.73 m[sup]2[/sup], calcium 9.4 (8.5-10 mg/dl), ionized calcium 5.4 (4.8-5.6 mg/dl), phosphorus 3.5 (2.5-4.6 mg/dl), and PTH 47 (12-65 pg/ml). Liver enzymes and alkaline phosphatase were normal. ANA was positive at 1:320; 25 hydroxyvitamin D was 30 (32-100 ng/ml). Coagulopathy screen was negative. Bone mineral density showed a femoral neck T-score of -2.8.[br]Mortality from calciphylaxis can be as high as 80%, due to sepsis from infected wound sites. Expert wound care is imperative. Other treatment options include sodium thiosulfate, cinacalcet, bisphosphonates, parathyroidectomy, or hyperbaric oxygen. Outcome data are very limited for these modalities in nonuremic calciphylaxis. With cessation of teriparatide therapy and aggressive wound care, our patient[apos]s lesions began improving. She was treated with intravenous zoledronic acid 5 mg and changed from warfarin to a low molecular weight heparin.[br]Conclusions: We report the first case of nonuremic calciphylaxis precipitated by teriparatide therapy. Our patient had multiple underlying predisposing factors including glucocorticoid therapy, warfarin use, and possible underlying coagulopathy or connective tissue disorder given her history of multiple deep venous thromboses and positive ANA. In such patients, alternative osteoporosis therapies may be preferred.[br][br]Nothing to Disclose: ES, DS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2212 61 461 SAT-371 PO46-02 Saturday 400 2012


401 ENDO12L_SAT-372 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Subungual Exostosis in a Patient Treated with Teriparatide Danae A Delivanis, Amit Bhargava, Pooja Luthra University of Connecticut Health Center, Farmington, CT Background: Recombinant PTH (Teriparatide) is commonly used for the treatment of severe osteoporosis. Rat studies using 3-60 times the approved human dose of teriparatide have shown an increased risk of osteosarcoma. Out of more than 430,000 patients treated by 2010, only 2 documented case reports of osteosarcoma exist. Subungual exostosis to our knowledge has not been reported.[br]Clinical Case: 54 yr old female presented with a 4 month history of significant pain and swelling in the medial side of her right thumb. There was no preceding history of trauma. Past history was significant for severe osteoporosis with multiple fractures, treated with teriparatide for 16 months. No family history of subungual exostosis. On examination, right thumb was swollen and tender to palpation. No erythema or signs of an infection were noted. X-ray imaging revealed a trabecular bony overgrowth consistent with subungual exostosis. Patient was treated with subungual excision and did well post-operatively. Pathology showed endochondreal bone formation with reactive atypia, consistent with osteocartilaginous exostosis.[br]Discussion: To our knowledge, this is the first report of a subungual exostosis in a patient treated with teriparatide. Subungal exostosis is a benign growth of bone that arises in the distal phalanx, under or adjacent to the nail bed. Differential diagnoses include verruca, a pyogenic granuloma or carcinoma of the nail bed. It affects women and men in a 2:1 ratio. The most common site of manifestation is the medial aspect of the great toe. Less commonly, it is seen in the index and middle fingers. Risk factors include trauma or inherited conditions such as multiple exostoses syndrome and multiple exostoses-mental retardation syndrome. The pathophysiology behind this growth is not clearly understood, but lesion has base of trabecular bone with a proliferating fibro cartilaginous cap. Teriparatide itself is known to stimulate the trabecular bone formation. Hence, an association between the use of teriparatide and the development of subungal exostosis cannot be excluded.[br]Conclusion: This is the first case hypothesizing an association between teriparatide and subungal exostosis. Subungal exostosis must be kept in the differential of any patient on teriparatide, presenting with an outgrowth on the distal aspect of their phalanx. Further studies delineating the relationship between teriparatide and subungal exostosis are needed.[br][br]Nothing to Disclose: DAD, AB, PL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1269 61 462 SAT-372 PO46-02 Saturday 401 2012


402 ENDO12L_SAT-373 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Atypical Femoral Fractures: Radiographic and Histomorphometric Features in 9 Patients Aliya Aziz Khan, Adil Hasan Zaidi, Nazir Khan McMaster University, Hamilton, Canada Purpose: This study describes characteristics and histomorphometric and radiographic features of atypical femoral fractures (AFF) as seen in 9 cases referred for evaluation.[br]Methods: All patients referred for evaluation of AFF were reviewed. Patients meeting the ASBMR criteria for AFF were further evaluated and tetracycline labelled bone biopsies were completed. Radiographs were reviewed by a musculoskeletal radiologist.[br]Results: All fracture lines were transverse or short oblique with thickened cortices. We report 9 cases of AFF in patients on long term bisphosphonate (BP) therapy. 6 of 7 fractures occurred without a fall or direct trauma to the femur with 1 case occurring after a fall from standing height. All patients were female; average age was 67 years (range 54-80 years). 2 of the 9 cases were of Chinese descent, 2 were East Indian with 3 being Caucasian. Average BP durations of us was 8.5 years (range 7-14 years). 5 of 9 patients were on alendronate alone, 2 patients were on risedronate. 1 patient had received 18 months of teriparatide, 3 years prior to AFF and had received a total of 10 years of BP use prior to teriparatide. Prodromal thigh or groin pain was seen in 5 of the 9 patients for 3 to 12 months prior to fracture. Proton pump inhibitor use was present in 1 patient for the previous 2 years. 1 patient was on prednisone for rheumatoid arthritis.[br]Rheumatoid arthritis was present in 2 cases. Diabetes was not present in any of the 9 cases. Bilaterality occurred in 1 patient with the second AFF occurring 1 month after the first AFF. Renal function was well preserved with estimated glomerular filtration (eGFR) rates [gt] 60 ml/min in all cases. 25 hydroxy Vitamin D levels were [gt]75 nmol/L in 5 cases. 2 cases had mild Vitamin D insufficiency ([gt]50 nmol/L).[br]Summary: A large number of the AFF occurred in women of Asian descent (4 of 9). 6 of the 9 AFF occurred in the absence of a fall. Prodromal pain was commonly seen. Proton pump inhibitors were used in only 1 patient. Histomorphometric features included evidence of mineralization abnormalities and decreased bone formation.[br]Conclusion: AFF in association with long term BP use are being seen in a disproportionately large number of Asian women. Further evaluation of all AFF with identification of predisposing key clinical risk factors is needed. Improved understanding of the pathophysiology may be gained with further histomorphometric data in larger numbers of patients.[br][br]Nothing to Disclose: AAK, AHZ, NK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 65 61 463 SAT-373 PO46-02 Saturday 402 2012


403 ENDO12L_SAT-374 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Transient Osteoporosis of Pregnancy Misdiagnosed as Osteomyelitis M Omar Salim, Mujahid Salim, Abdul Nuristani, Khalid Salim, Mahmood Shahlapour, Mohamad H Horani Mercygilbert Hospital, Gilbert, AZ; Good Samaritan Hospital, Phoenix, AZ; Alsham Endocrinology, Chandler, AZ [bold]Introduction[/bold]:[br]Transient osteoporosis is a condition characterized by bone pain along with X-ray evidence of osteoporosis and MRI signs of inflammation. This condition usually resolves itself within few months.[br][bold]Case Study[/bold] :[br]A 36 year old Hispanic female with no significant past medical history was admitted to the hospital for impending child birth. She had also inguinal pain that started during her third trimester, which spread down to her mid thigh. The patient denied any fever, no falls or trauma.[br]On Physical Exam: Patient had significant tenderness on the right inguinal area and right thigh. There was some erythema, but no skin breakdown. She had some tenderness on the left side as well.[br]Her lab results revealed calcium levels of 8.1 mg/dL. Vitamin D-25 levels were 6 ng/mL (30-100 ng/mL). Vitamin D-1,25 levels were 12 pg/mL (15-75 pg/mL). PTH levels were 32 pg/mL (11-67 pg/mL). The ESR was 76. WBC was 6.2.[br]An MRI of the femur and thigh showed results consistent with diffuse moderate cellulitis of the right thigh,and left Knee. After initial osteomyelitis concern, patient was started on antibiotics and asked to abstain from breastfeeding. After infectious disease consult, there was felt to be no real evidence of osteomyelitis. So antibiotics were discontinued. The patient underwent a biopsy of the right distal femur, during which, a fracture occurred and required surgery. Bone biopsy showed nonspecific reactive reparative changes. the culture was negative. After discussing the case with the radiologist, it was concluded that MRI findings that resembles infection could also be seen in transient osteoporosis of pregnancy.[br][bold]Discussion:[br][/bold]Transient osteoporosis is not a fully understood condition, but is distinguished by self-limited pain and radiographically evident osteopenia. Transient osteoporosis typically occurs in middle aged women in the third trimester of pregnancy or men. There is a spontaneous onset of pain, that usually progresses over several weeks. Transient osteoporosis can manifest as pain in hips, groin, buttocks, or thighs. A feared complication of this condition is hip fracture. Clinical improvement occurs over several weeks to months without specific treatment. The role of vitamin D deficiency in our case as a predisposing factor for transient osteoporosis is yet to be known.[br][bold]Conclusion:[br][/bold]Peripartum women with hip pain and without supportive medical history should have transient osteoporosis included in the differential diagnosis.[br][br]Nothing to Disclose: MOS, MS, AN, KS, MS, MHH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 66 61 464 SAT-374 PO46-02 Saturday 403 2012


404 ENDO12L_SAT-375 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Cinacalcet Use in X-Linked Hypophosphatemic Rickets Complicated by Tertiary Hyperparathyroidism and Hypercalcemia Srikanth Mitta, Amal Shibli-Rahhal University of Iowa Hospitals and Clinics, Iowa City, IA [bold]Introduction:[/bold] X-linked hypophosphatemia (XLH) is caused by a loss of function mutations in the endopeptidase PHEX, which results in increased levels of FGF23. FGF23 suppresses 1-[alpha] hydroxylation of vitamin D and inhibits renal tubular reabsorption of phosphate, leading to hypophosphatemia, rickets and growth retardation. Treatment consists of oral phosphate and calcitriol, but this may be complicated by secondary and tertiary hyperparathyroidism (HPT).[br][bold]Clinical case:[/bold] We report a 70-year-male with XLH treated with calcitriol and phosphate for most of his adult life. In 2004 he was found to have a PTH of 249 pg/ml (10-65), serum calcium of 9.6 mg/dl (8.5-10.5), phosphorus of 2.6 mg/dl (2.5-4.5) and creatinine of 0.8 mg/dl (0.7-1.4). He was on potassium phosphate (K-Phos) 500 mg tid and calcitriol 0.5 mcg daily. K-Phos was decreased to 500 mg bid to maintain a serum phosphorus of 2-2.5 mg/dl. A lower dose of K-Phos was not tolerated due to fatigue and bone pain. PTH decreased to 99 pg/ml in 6 months and remained stable for 2 years with normal serum calcium. Subsequently both his PTH and calcium started increasing and continued to increase despite discontinuation of calcitriol. As his serum calcium reached 12 mg/dl with PTH 405 pg/ml, parathyroidectomy was considered but was not possible as the patient could not extend his neck due to a previous cervical spine fusion surgery. Cinacalcet 30 mg bid was initiated and the calcium decreased to 9.6 mg/dL in 2 weeks and has remained normal since. His PTH improved from 405 pg/mL to 295 pg/mL within 3 months.[br][bold]Discussion:[/bold] Oral phosphate and calcitriol are effective at raising the serum phosphate and healing rickets and osteomalacia in XLH. However, oral phosphate causes secondary HPT by acutely raising the plasma phosphate, which binds plasma calcium, causing transient hypocalcemia and by directly stimulating the release of PTH from the parathyroid glands. Long-term stimulation of the parathyroid glands can result in hyperplasia, autonomous hyperfunction and hypercalcemia due to tertiary HPT. Parathyroidectomy is the preferred treatment for these patients. When surgery is not possible, calcimimetics can correct the hypercalcemia and reduce serum PTH by directly targeting the calcium sensing receptor on the parathyroid glands.[br][bold]Conclusion:[/bold] Cinacalcet may correct hypercalcemia in patients with XLH who develop tertiary HPT and in whom surgery is not possible.[br][br]Nothing to Disclose: SM, AS-R 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1959 61 465 SAT-375 PO46-02 Saturday 404 2012


405 ENDO12L_SAT-376 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Heart Failure in Hypophosphatemic Rickets: Lessons Learned from the Complications of High-Dose Phosphate Therapy Grace Sun, Ozan Suer, Melissa Li-Ng Cleveland Clinic Foundation, Cleveland, OH; Cleveland Clinic Lerner College of Medicine, Cleveland, OH [bold]Background: [/bold]Hypophosphatemic rickets (HR) is caused by excess phosphorus excretion which impairs bone mineralization. Complications of HR have decreased with the advent of vitamin D analogues used with oral phosphate. However, high phosphate doses, even with calcitriol use, can cause secondary hyperparathyroidism (HPT) by stimulating parathyroid hormone secretion, complexing with serum calcium and decreasing calcitriol via FGF-23 at the renal tubule. These mechanisms may lead to autonomous parathyroid (PT) function. We present a 34-year old male with HR who developed tertiary HPT with extensive heart calcifications causing heart failure due to inappropriate phosphate-only therapy as a child.[br][bold]Case: [/bold]Our patient was diagnosed with HR after presenting at 4 years old with delayed growth and leg bowing. Family history was negative for HR. He was placed on high-dose phosphate therapy (2-6 grams/day) with sporadic calcitriol use between 4-18 years of age. For 6 years total, he was on high-dose phosphate [italic]without[/italic] calcitriol. He subsequently developed nephrocalcinosis, heart valve calcifications requiring subsequent multi-valve replacements, severe calcific coronaries ([ge]90% occlusion of 3 major vessels), heart block, and cardiomyopathy from extensive endocardial/myocardial calcifications. He required a biventricular pacemaker and an implantable cardioverter defibrillator. Most of his cardiac interventions took place in his 20[apos]s. He was lost to endocrine follow-up for 15 years, and his secondary HPT progressed to tertiary HPT. He presented to our hospital for valve replacement, but his hospital course was complicated by decompensated heart failure and multisystem organ failure. His autopsy showed hyperplasia of the 3 identified PT glands.[br][bold]Conclusion: [/bold]Cardiac calcifications are a known complication of tertiary HPT when it occurs in patients with renal failure, but it is rarely seen in HR due to high phosphate therapy. Phosphate alone should never be used to treat HR; high doses, even with calcitriol, should be avoided. It is unclear what the [ldquo]safe[rdquo] oral phosphate threshold is to avoid HPT in these patients. Our case was an extreme example of cardiac complications from high-dose phosphate therapy and resultant tertiary HPT. It is important to be aware of high-dose phosphate effects and to consider parathyroidectomy for autonomous function, if needed. Our patient[apos]s critical illness precluded parathyroidectomy, but he would have benefited from surgery for his tertiary HPT.[br][br]Nothing to Disclose: GS, OS, ML-N 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 230 61 466 SAT-376 PO46-02 Saturday 405 2012


406 ENDO12L_SAT-377 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Vitamin D Deficiency (VDD) and Major Depressive Disorder (MDD): A Causal or Casual Association? Sonal Pathak, Mohammad Asim Nisar, David S Rosenthal Bayhealth Endocrinology, Dover, DE; St Lukes Roosevelt Hospital, New York, NY; Nassau University Medical Center, East Meadow, NY [bold]Background:[/bold] Vit D is associated with several metabolic, musculoskeletal and immunologic effects. Its psychosomatic effects have been speculated, but less studied. We present 3 cases of VDD in patients with MDD, with significant improvement in mood after repletion. We[apos]ll discuss controversies regarding effects of Vit D on mood and cognitive disorders and a potential role of Vit D replacement as an adjunct in management of MDD in Vit D deficient patients.[br][bold]Cases: [/bold]Patient #1 was a 42 y/o F treated for hypothyroidism with obesity and depression for 5 years. She was euthyroid on a stable dose of L-T4 with TSH 1.24[micro]IU/ml (0.45-4.50). In spite of optimization of antidepressant therapy, her depression had exacerbated over the preceding 6 months (*BDI 32). Patients #2 and #3 were females aged 58 and 66 years, being treated for DM2 and also suffered from depression (*BDI 26 and 21 respectively). VDD was suspected based on risk factors/symptoms. Presenting 25(OH)VitD levels were low (normal 30-85 ng/ml) at 12, 8.9 and 14.5ng/ml respectively. After Vit D repletion, 25(OH)Vit D levels became sufficient at 32, 34.6, and 38ng/ml respectively. Other medications and environmental factors were unchanged. Depressive symptoms were reported to be significantly improved by all patients (*BDI 12, 8 and 16 respectively).[br][bold]Discussion:[/bold] Vit D receptors and [alpha]-hydroxylases have been mapped throughout the brain, including the amygdala/limbic system, where behavior/emotions are regulated. Vit D is also involved in release of neurotransmitters like serotonin/dopamine in brain. Association of VDD with dementia [amp] schizophrenia has been well studied. However, data from epidemiological studies and a few randomized controlled trials on relationship between Vit D status and depression has been equivocal, although suggestive of a possible causal association. Effects of Vit D replacement in deficient patients with MDD has been less studied, with a few reports suggesting a significant positive effect, but limited by confounding variables.[br][bold]Conclusion:[/bold] Detection and treatment of VDD in patients with depression may be an easy and cost-effective adjunct to mainstream therapies; however, more research is needed to definitively confirm these findings.[br]*Beck Depression Inventory score (BDI): 0[ndash]9 minimal depression; 10[ndash]18 mild depression; 19[ndash]29 moderate depression; 30[ndash]63 severe depression.[br][br]1. Milaneschi et al, JCEM 95(7), July 2010. 2. Ganji et al, Int Archives of Medicine, 2010, 3:29. 3. Bertone-Johnson, Nutr Rev. August 2009; 67(8).[br][br]Nothing to Disclose: SP, MAN, DSR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 830 61 467 SAT-377 PO46-02 Saturday 406 2012


407 ENDO12L_SAT-378 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Osteomalacia and Severe Hypocalcemia after Bariatric Surgery and Partial Parathyroidectomy Luciana Valadares Ferreira, Lilian Araujo Caetano, Denise Yumi Eimori, Raquel Resende Silva, Ricardo Ayello Guerra Hospital do Servidor P[uacute]blico Municipal, S[atilde]o Paulo, Brazil Background: Gastric bypass surgery procedures are a common practice for the treatment of morbid obesity. Metabolic bone disease is a well-documented long-term complication of obesity surgery. It is often undiagnosed, or misdiagnosed, because of lack of physician and patient awareness. Abnormalities begin shortly after gastrointestinal bypass operations and include osteoporosis, osteomalacia and secondary hyperparathyroidism (HPT). Despite the great risk of hypocalcemia these patients still have the defense mechanism that is secondary HPT. Clincal Case: We report a case of a 45 year-old woman who underwent a biliopancreatic diversion (BPD) surgery in 2006. After BPD, the patient lost 67Kg (from a BMI of 52,9 kg/m2 to a BMI of 31,7 kg/m2). Two years later she exhibited progressive back pain and was diagnosed with vertebral body[apos]s fracture. At that time: BMD Tscore L1-L4 -3,2, ionic calcium (Cai) 1,0mmol/l (RV: 1,0-1,32), phosphorus (P) 3,1mg/dl (RV: 2,5-4,9), alkaline phosphatasis (AP) 358U/l (RV: 40-150) and PTH 849pg/ml (RV: 12-65), suggestive of osteomalacia. She was diagnosed with osteoporosis and primary HPT and had two parathyroid glands surgically removed. Since then she started with frequent cramps, extremities and perioral paresthesia, with persistently high PTH, hypocalcemia and low serum 25-hydroxivitamine D (25OHD). Osteomalacia was diagnosed on the basis of her biochemical profile - serum calcium (Ca) 7,42mg/dl (RV 8,9-10), urinary calcium 38mg/24h, AP 262U/l, P 3,7mg/dl, PTH 507pg/ml and 25OHD 10,4ng/ml (RV 30-100). The severity of the hypocalcemia was exacerbated after the parathyroidectomy. At this time she was taking alendronate, calcium carbonate and calcitriol. The bisfosfonate was interrupted and the dose of supplementary calcium and vitamin D was progressively increased until calcium citrate 9g/day, colecalciferol 50.000U/day and calcitriol 1mcg/day. That leads to no improvement of the biochemical profile that shows 25OHD 5,76ng/ml. Parenteral ergocalciferol (600.000 units) was administered with an improvement of 25OHD levels (to 23,1ng/ml after 1 month). Conclusion: The reported case shows the importance of the correct analysis of biochemical profile of calcium in patients after bariatric surgery to avoid possible diagnostic errors that lead to iatrogenic worsening of bone metabolic disease.[br][br]Nothing to Disclose: LVF, LAC, DYE, RRS, RAG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1030 61 468 SAT-378 PO46-02 Saturday 407 2012


408 ENDO12L_SAT-379 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Two Different Cases of Vitamin D Deficiency in Patients after Surgery of Gastric Bypass Maria Prokopiou, Lia Zapanti, Nektarios Benetatos, Maria Christou, Athanasios Karahalios Laiko Hospital, Athens, Greece Introduction[br]Vitamin D deficiency is more common after bariatric surgery due to malabsorption.[br]Two different cases of patients having undergone surgery of gastric bypass, with Vitamin D deficiency will be presented here.[br]Case 1: A 49-year-old woman was presented to us for an osteoporosis check walking with the use of a jib-boom.[br]Medical History: Bariatric surgery(loss 140 kg in 8 months), menopause, spontaneous fracture of L1 vertebra. Present illness: Muscle and Bone pain at the lumbar and thoracic vertebral column difficulty with walking for the last 4 years. On physical examination: She exhibited lumbar kyphosis with thoracic and lumbar percussive pain and was depressive. Her body height was 155 cm (previously 166 cm) and her weight was 65 Kgr. She had muscle atrophy and symmetrical loss of proximal muscle strength. Laboratory evaluation: Serum: Ca=8.1mg/dl,P=3.2 mg/dl,Alb=4.1g/dl, 25(OH)VIT D=3.1 ng/ml, PTH=178 pg/ml, TSH=3.2 mU/L, ALP=95 U/L[br]Bone mineral density measurement:[br]BMD T-score[br]LWK (L1[ndash]L4) 0.578 -5[br]HIP 0.529 -3.9[br]The patient was prescribed 6000 IU/day Vitamin D and 1500 mg/day Calcium carbonate. After correcting serum Ca, she was also prescribed anti-osteoclastic therapy. After 5 months of administration, she exhibits improvement in underlying symptoms and clinical manifestation. However, Vitamin d remained low.[br]Case 2: A 42-year-old man was presented to us having difficulties with walking and sensitivity disorders in low extremities (burning sensations).[br]Medical History: Bariatric surgery of gastic bypass(loss 70 kg in 8 months).Present illness: Sensibility disorders of the legs, muscle weakness in the legs. On physical examination: He had proximal muscle weakness, tendon hyporeflessia. He was walking with small steps, diminished superficial and profundus sensibility in the legs and was depressive.[br]Laboratory evaluation: Serum: Ca=9.5 mg/dl, P=3.7 mg/dl, Alb=4.4 g/dl, 25(OH)Vit D=8 ng/ml, PTH= 90 pg/ml, TSH=2,1 mU/L, ALP=93 U/L. [br]Bone mineral density measurement:[br]BMD T-score[br]LWK (L1[ndash]L4) 0.947 -2[br]HIP 0.952 -1.8[br]The patient was prescribed 6000 IU/day Vitamin D. After 6 months of administration, there was an increase in 25(OH)VitD level (25(OH)VitD=24mg/dl) and further subjective(muscle weakness, depression) and objective improvement.[br]Conclusion[br]Vitamin D deficiency can manifest itself in a variety of ways including bone metabolism disease, neuromuscular disorders and depression.[br][br]Nothing to Disclose: MP, LZ, NB, MC, AK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1281 61 469 SAT-379 PO46-02 Saturday 408 2012


409 ENDO12L_SAT-380 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Post Zoledronic Acid (ZOL) Hypocalcemia in a Vitamin D Replete Roux-en-Y Gastric Bypass (RYGB) Patient Liviu George Danescu, Ruban Dhaliwal, Arnold M Moses State University of New York (SUNY) Upstate Medical University, Syracuse, NY Background: The incidence of hypocalcemia after infusion of ZOL can be markedly decreased by assuring adequate calcium intake and optimizing pre-infusion vitamin D levels. In patients who have had RYGB surgery, the duodenum, a major source of active calcium absorption is bypassed, thus perhaps predisposing to post-infusion hypocalcemia.[br]Clinical case: A 64 year-old male had RYGB 3 years prior to referral for evaluation and treatment of Paget[apos]s disease. He had no gastrointestinal complaints and no bone pain. On presentation: PTH 84 pg/ml (15-65), alkaline phosphatase 143 U/L (n[lt]104), BSAP 33.7 ug/L (n[lt]20.1), serum NTX 34.2 nM BCE (n[lt]24.2), 25-OH vitamin D 36 ng/ml (30-100), calcium 8.8 mg/dL (8.8-10.2), albumin 4.2 g/dL (3.4-4.8), phosphorus 3.1 mg/dL (2.7-4.5), magnesium 1.5 meq/L (1.3-2.1), urine calcium 34.5 mg/24h (n[gt]100) with creatinine 1800 mg/24h. X-ray, CT and bone scans were consistent with Paget[apos]s disease of the right proximal femur. His medications included ergocalciferol 50,000 IU biweekly and 500 mg elemental calcium as carbonate three times a day. Dietary calcium intake was minimal.[br]Ergocalciferol was increased to weekly and three months later he was treated with 5 mg IV ZOL. His pretreatment 25-OH vitamin D level was 46 ng/ml, calcium 9.4 mg/dl, PTH 111 pg/ml and 24 h urine calcium 28.8 mg with creatinine 1600 mg. Three days post infusion calcium decreased to 7.7 mg/dl without symptoms. Elemental calcium as citrate 315 mg three times daily was added. His two-week post infusion calcium level normalized to 8.6 mg/dl while PTH rose to 189 pg/ml and phosphorus decreased to 2.1 mg/dL. His 24 h urine calcium remained low at 31.1 mg. The average of four calcium levels over 2 weeks post infusion was 8.1 mg/dl compared to an average of 9.1 mg/dl for four calcium levels over 3 months prior.[br]Conclusion: This patient with a gastro-duodenal bypass procedure developed hypocalcemia after infusion with ZOL even though his vitamin D level was normal. The ZOL apparently unmasked a pre-infusion calcium deficit which caused a further elevation of PTH which may have lowered his serum phosphorus. His 24h urine calcium excretion remained low. The findings in this case indicate that before ZOL is administered to patients who have had a Roux-en-Y procedure, in addition to normalizing vitamin D levels an effort should be made to normalize PTH, urine calcium or both to prevent hypocalcemia.[br][br]Nothing to Disclose: LGD, RD, AMM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 861 61 470 SAT-380 PO46-02 Saturday 409 2012


410 ENDO12L_SAT-381 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) An Interesting Case of Peripheral Vascular Disease, Vascular Reperfusion and Subsequent Development of Pain Due to Paget Disease of Bone Sunna Kwun, Joseph R Tucci Rhode Island Hospital, The Warren Alpert Medical School at Brown University, Providence, RI; Roger Williams Medical Center, Boston University School of Medicine, Providence, RI Paget[apos]s disease of bone is a focal disorder of chaotic bone remodeling with an increase in osteoblastic and osteoclastic activity that results in disorganized and structurally abnormal bone in one or more skeletal sites. Pagetic bone is of poor quality, enlarged, and susceptible to deformation and fracture. Pagetic bone and bone marrow are hypervascular and, as a result, skin overlying pagetic bone (e.g. tibia or skull) is often warmer than unaffected skeletal sites. Paget[apos]s disease of bone has been associated with an increase in vascular calcifications and an increase in cardiovascular disease particularly in those with extensive disease.[br]79-year-old-woman with a history of coronary artery heart disease and recent finding of a T5 compression fracture following a motor vehicle accident was hospitalized for evaluation of right lower extremity claudication. Angiography demonstrated a focal complete occlusion of the distal right femoral and popliteal arteries. A self-expanding stent was placed in the distal femoral and popliteal arteries. Approximately 48 hours after the procedure, the patient developed severe right lower leg pain. Sciatic nerve impingement and osteomyelitis were considered and excluded as causative. She was discharged on pain medication to a rehabilitation center. Two weeks later, she was again hospitalized for excruciating pain in the right lower leg with an inability to bear weight. On Cardiac evaluation, a reperfusion complication was excluded. She was seen by Pain Management and Neurology and was treated with a narcotic and neuroleptic agent. On endocrine evaluation, anterior bowing and a marked increase in warmth was noted over the right tibia as compared with the contralateral tibia. Serum alkaline phosphatase level was increased at 127 IU/ml (normal 25-100 IU/ml). A bone scan revealed intense uptake in the proximal two-thirds of the right tibia consistent with Paget[apos]s disease of bone. X-rays revealed thickening and coarsening of the trabecular pattern involving the proximal two-thirds of the tibia with slight anterior bowing of the tibia.[br]In summary, this patient[apos]s development of severe pain following reperfusion of distal femoral and popliteal arteries is in keeping with the known and aforementioned hypervascularity of pagetic bone. The finding of increased warmth over an area of skeletal deformation should always raise the possibility of Paget[apos]s disease of bone.[br][br]Nothing to Disclose: SK, JRT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 316 61 471 SAT-381 PO46-02 Saturday 410 2012


411 ENDO12L_SAT-382 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Worsening of Intracranial Hypertension and Prolonged Hypocalcemia Following Treatment of Paget Disease of the Skull with Zoledronic Acid Bruno Ferraz-de-Souza, Helena Nicolielo, Cristiane N Lauretti, Regina M Martin, Pedro Henrique S Correa Hospital das Clinicas, University of S[atilde]o Paulo School of Medicine, S[atilde]o Paulo, Brazil Background: Skull involvement in Paget[apos]s disease leads to hearing loss, tinnitus, and, in severe cases, intracranial hypertension (IH), prompting bisphosphonate treatment. Intravenous zoledronic acid (ZA) has emerged as an effective and safe treatment option for patients with Paget[apos]s, leading to sustained remission and improved quality of life. Rarely, symptomatic hypocalcemia might ensue from ZA infusion, particularly when vitamin D deficiency or hypoparathyroidism are present. Potentially direct adverse effects of bisphosphonates on intracranial pressure have not been reported.[br]Clinical Case: A 61-year-old female presented with a 2-year history of facial asymmetry with progressive hearing impairment and holocranial headache. A pronounced left temporal bone deformity with increased local temperature was evident. She had not been previously treated. Total serum Ca levels were normal (9.5 mg/dl; NR 8.6-10.2) with upper normal levels of PTH (71 pg/ml; NR 16-87) and low 25-OHD levels (13 ng/ml; NR[gt]20). Serum alkaline phosphatase was markedly increased (2,866 U/L; NR 35-104) and bone scan showed extensive pagetic involvement of the skull, confirmed by radiography. Head CT and MRI revealed IH (supratentorial hydrocephalus, tonsillar herniation), platybasia and basilar invagination. Based on normal fundoscopy and absence of clinical signs of IH, initial neurosurgical evaluation forwent emergency treatment and sanctioned bisphosphonate therapy. Vitamin D and Ca were optimized, and 15 d later the patient was treated with 4 mg intravenous ZA. Twelve hours after the infusion, she became confused, agitated and disoriented and developed urinary incontinence. Serum Ca was 8.6 mg/dl and cortical sulci became effaced on CT indicating increased intracranial pressure. Over the next days, she developed frank hypocalcemia (nadir 7.0 mg/dl) requiring IV Ca infusion and calcitriol. Neurological status returned to normal and the patient was discharged 4 d later with prescribed oral Ca supplementation and calcitriol. Six months later she still required calcitriol to maintain normal calcemia and remained neurologically stable.[br]Conclusion: Following ZA infusion, our patient with extensive Paget[apos]s disease of the skull experienced an acute worsening of previously asymptomatic IH and prolonged hypocalcemia. While it is unclear whether ZA affected IH directly or, most likely, through hypocalcemia, we believe this report may serve as an alert for the future management of similar cases.[br][br]Nothing to Disclose: BF-d-S, HN, CNL, RMM, PHSC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1222 61 472 SAT-382 PO46-02 Saturday 411 2012


412 ENDO12L_SAT-383 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Recurrent Pleural Effusion after Zoledronic Acid in a Patient with Fibrous Dysplasia Tulsi Sharma, Jennifer J Kelly, Arnold M Moses State University of New York (SUNY) Upstate Medical University, Syracuse, NY Introduction: Multiple serious adverse effects have been attributed to bisphosphonate use; some resulting from off-label uses, but others seen at the recommended dosage and infusion time.[br]Case: We present the case of a 41 year-old male with polyostotic fibrous dysplasia since 1986. He had increasing bone pain for which he received IV zoledronic acid 5mg infusion in April 2011. He complained of progressive dyspnea over the next few weeks. Workup revealed large bilateral exudative pleural effusions. He underwent a therapeutic thoracentesis and left drain placement which was removed over the next 2 days.[br]His symptoms recurred the following month; he was again found to have massive pleural effusions and underwent thoracentesis of more than 1800 ml fluid. Since that time, his effusions have been recurrent requiring repeated thoracentesis. His course over the last few months has been complicated by development of pleural fibrosis and trapped lung as a result of his effusions. He has recently undergone a right sided VATS(video-assisted thoracic surgery) with parietal pleurectomy and decortication. Pleural biopsy showed evidence of fibrotic adhesions. Rib lesion biopsy showed fibrous dysplasia without any evidence of malignancy. Despite a complete and exhaustive work-up, no specific cause has been identified for this effusion. Is this related to the use of zoledronic acid?[br]Discussion: Exudative pleural effusion has wide differential and thinking out of the ordinary can sometimes help if no obvious cause is found. This is especially the case for newer drugs with which we do not have long term data and experience.[br]Our patient has a long history of fibrous dysplasia but did not have any prior pleural effusions. The occurrence of recurring effusions with recent use of zoledronic acid points to a diagnosis of this being a rare medication side-effect. A detailed search of literature shows that a few cases have been reported to the FDA after use of zoledronic acid but none have been published. Dyspnea is a listed side effect occurring in 22-27% patients but the etiology is not elucidated.[br]As the clinical indications for bisphosphonate use continue to expand, it is important for clinicians to prevent, recognize, and manage any possible complications effectively and expeditiously. This case raises awareness to be vigilant for the possibility of a pleural effusion after the use of zoledronic acid as dyspnea is a frequently reported complication.[br][br](1)Abrahamsen B. Curr Opin Rheumatol. 2010 Jul;22(4):404-9. (2)Diel IJ, et al. J Support Oncol. 2007 Nov-Dec;5(10):475-82. (3)Mansoori LS, et al. Endocr Pract. 2010 Sep-Oct;16(5):851-4. (4) http://www.uptodate.com/contents/zoledronic-acid-drug-information.[br][br]Nothing to Disclose: TS, JJK, AMM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1154 61 473 SAT-383 PO46-02 Saturday 412 2012


413 ENDO12L_SAT-384 POSTER SESSION: Metabolic Bone Diseases (1:30 PM-3:30 PM) Non-Uremic Calciphylaxis: Rare Disease, Common Outcome J Barr Biglane, Sarah E French, Jose Subauste University of Mississippi Medical Center, Jackson, MS; GV (Sonny) Montgomery Veterans Affairs Medical Center, Jakcson, MS Background: Calciphylaxis, or calcific uremic arteriolopathy, occurs in 1-4% of patients with ESRD and is associated with high mortality rate of 60-80%, usually due to sepsis1. Other risk factors include female gender, white race, obesity, systemic steroid use, warfarin use, and raised serum calcium phosphate product. There is no standardized treatment but cases with response to bisphosphonates and sodium thiosulfate have been reported2.[br]Clinical case: A 32 year-old white female with steroid-dependent rheumatoid arthritis, chronic bronchopleural fistula from recurrent pneumothoraces presented with painful purpuric lesions and ecchymoses on bilateral lower extremeties. She denied any warfarin use or history of renal failure. On physical exam, she had painful, ecchymotic, non-blanching lesions on her right lower leg, left ankle and abdomen, measuring 0.5cm to 3 cm. Laboratory confirmed normal calcium, phosphorus, and renal function. Her skin lesions slowly erupted and multiplied into large stellate-shaped ulcerations with fibrinous exudate and no obvious granulation tissue. A few developed black, necrotic centers. Lesions also developed on her buttocks, thigh, hips, lower abdomen, and left arm. A biopsy of a representative lesion showed calcification of the small and medium-sized blood vessels with occlusion and necrosis as well as necrotic foci in subcutaneous tissue and dermis[mdash]consistent with calciphylaxis. Treatment with sodium thiosulfate and pamindronate was initiated in addition to aggressive pain control and wound debridement. However, despite several weeks of treatment, she had no improvement in her lesions. Given her overall health, she chose to stop further treatment and began hospice care.[br]Conclusion: While there is no standard therapy for calciphylaxis, surgical debridement often becomes necessary and was associated with prolonged survival in one study3. Successful treatment with sodium thiosulfate has been described in both a patient with chronic renal failure requiring dialysis4 as well as a patient with normal renal function2 by either chelating calcium from involved tissues or its antioxidant activitiy1. Bisphosphonates (etidronate, pamidronate, and ibandronate) have used in five patients on hemodialysis with good outcomes1. Ours is one of the only cases in which both therapies were tried but no response to treatment was seen. The mortality of calciphylaxis remains high with a 1-year survival rate of just 45.8%3.[br][br]1. Raymond CB, Wazny LD. Sodium thiosulfate, bisphosphonate, and cincalcet for treatment of calciphylaxis. Am J Health Syst Pharm,2008;65:1419-20. 2. Hackett BC, McAleer MA, Sheehan G, et al. Calciphylaxis in a patient with normal renal function: response to treatment with sodium thiosulfate. Clin Exp Dermatol,2009;34:39-42. 3. Weenig RH, Sewell LD, Davis MD, et al. Calciphylaxis: natural history, risk factor analysis, and outcome. J Am Acad Dermatol,2007;56:569-79. 4. Cicone JS, Petronis JB, Embert CD, et al. Successful treatment of calciphylaxis with intravenous sodium thiosulfate. Am J Kidney Dis, 2004;43:1104-8.[br][br]Nothing to Disclose: JBB, SEF, JS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1923 61 474 SAT-384 PO46-02 Saturday 413 2012


414 ENDO12L_SAT-386 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Dissociation between LPS-Induced Changes in Type 2 Deiodinase and OATP1c1 Expression in the Brain Suggests T3 Action Is Localized to the Meningeal Compartment Gabor Wittmann, Ronald M Lechan Tufts Medical Center, Boston, MA; Tufts University School of Medicine, Boston, MA Recently, we demonstrated that the administration of bacterial lipopolysaccharide (LPS) to rats induces marked, [italic]de novo[/italic] transcription of the type 2 deiodinase (D2) gene in the leptomeninges and choroid plexus, resulting in a 50-100-fold increase in D2 enzymatic activity in these tissues within 9h (1). At the same time, the mRNA of organic anion transporting polypeptide (OATP1c1), a high affinity T4 transporter, is virtually absent from endothelial cells in the brain (2). Since elevated T3 levels are known to downregulate OATP1c1 in blood vessels, we hypothesized that the reduction in OATP1c1 expression by LPS may be secondary to increased T3 levels generated by D2 at the blood-brain/cerebrospinal fluid barrier. Therefore, we studied the time course of D2 and OATP1c1 mRNA expression by [italic]in situ[/italic] hybridization following LPS administration, expecting that reduction in OATP1c1 mRNA will follow upregulation of D2 mRNA. Contrary to expectations, OATP1c1 mRNA was markedly reduced in blood vessels as early as 2h following LPS, and absent by 4h, whereas D2 mRNA in the leptomeninges was not detected at 2h, low by 4h, and maximally increased by 9h. As another index of T3 concentration in brain tissue, the expression of type 3 deiodinase (D3) was studied, an enzyme known to be upregulated by thyroid hormone. Neuronal D3 mRNA was not significantly changed in the brain even 9h after LPS administration. These data suggest that the marked increase in D2 expression in the leptomeninges and choroid plexus may not significantly affect T3 levels in the brain neuropil. Rather, the rapid, LPS-induced downregulation of OATP1c1 in brain endothelial cells may result in reduced thyroid hormone levels in neuronal tissue. Since thyroid hormone has been linked to reduced proinflammatory cytokine levels and attenuated inflammatory tissue damage (3), we hypothesize that the relatively late induction of D2 in the leptomeninges by LPS may be an important component of a negative feedback response that restrains proinflammatory cytokine expression by increasing the local meningeal concentration of T3.[br][br](1) Wittmann et al., Endocr Rev 2010; 31: Meeting Abstracts OR41-2. (2) Wittmann et al., Endocr Rev 2011; 32: Meeting Abstracts P1-659. (3) Barca-Mayo et al., PNAS 2011; 108: E1321.[br][br]Sources of Research Support: NIH DK37021.[br][br]Nothing to Disclose: GW, RML 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2132 62 475 SAT-386 PO38-01 Saturday 414 2012


415 ENDO12L_SAT-387 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Analysis of Cellular Localization and Iodide Transport of Pendrin (SLC26A4) Mutants Involving the STAS Domain Aigerim Bizhanova, Peter Kopp Northwestern University, Feinberg School of Medicine, Chicago, IL Pendrin (SLC26A4) is a hydrophobic multifunctional anion transporter expressed at the apical membrane of thyrocytes, where it is thought to be involved in mediating iodide efflux. Biallelic mutations in the SLC26A4 gene lead to Pendred syndrome, an autosomal recessive disorder characterized by sensorineural deafness, goiter and a partial defect in iodide organification. Like other members of the SLC26A family, pendrin contains a so-called STAS (sulfate transporter and antisigma factor antagonist) domain in its carboxy-terminus. This domain shares similarity with the bacterial anti-sigma factor antagonists. The STAS domain of pendrin is thought to be composed of a proximal (residues 535 to 573) and distal (residues 654 to 729) domain. While the functional role of the STAS domain needs further characterization, it has been suggested that it is involved in nucleotide binding and/or interactions with other proteins. We characterized pendrin mutants lacking the proximal, the distal, or both parts of the STAS domain and determined the effect of these mutations on cellular localization and the ability to transport iodide.[br]A GFP-tagged pendrin mutant lacking the proximal part of the STAS domain shows membrane localization that is similar to the expression pattern of wild type pendrin in transiently transfected rat kidney epithelium (Ptk2) cells. Moreover, time-lapse imaging of living PtK2 cells demonstrates that a considerable amount of the mutant rapidly translocates to the plasma membrane in response to activation of the protein kinase A pathway by forskolin. In contrast, GFP-tagged mutants lacking either the distal part, or both the proximal and the distal elements of the STAS domain, are located in intracellular compartments and fail to translocate to the plasma membrane in response to forskolin. Consistent with the localization studies, the functional analyses indicate that deletion of the distal part of the STAS domain results in complete inability of the protein to mediate iodide efflux. Deletion of the proximal part of the STAS domain does not affect the ability of the protein to mediate iodide efflux.[br]In conclusion, the presence of the distal part of the STAS domain of pendrin is required for membrane targeting and its ability to mediate basal and forskolin-stimulated iodide efflux.[br][br]Nothing to Disclose: AB, PK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2157 62 476 SAT-387 PO38-01 Saturday 415 2012


416 ENDO12L_SAT-388 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Low Frequency of TPO Gene Mutations in Patients with Iodide Organification Defects Ester Saraiva Brust, Cristine Barbosa Beltrao, Suemi Marui Disciplina de Endocrinologia-Faculdade de Medicina-Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Defects in thyroid hormones synthesis account for 15% of congenital hypothyroidism (CH)and correspond to genetic defects mainly defective organification of iodide (I)due to defects in the peroxidase enzyme (TPO) or in H2O2 generating system. The association of an organification defect with sensorineural deafness is known as Pendred syndrome. TPO defect is the most frequent and is commonly associated with an autosomal recessive form of inheritance. Objective: Search for mutations in TPO in patients with suggestive I organification defect. Methods: We evaluated 40 patients with permanent CH from the Association for Parents and Friends of Disabled Individuals (APAE) of Sao Caetano outpatient clinic, and referred to our Service previously described (1). All patients had hormonal evaluation, color-Doppler ultrasound (CDUS), radionuclide scan with 131I and 99Tc Pertechnetate. Perchlorate discharge (PCIV) test was performed only in patients with normal or enlarged thyroid detected by CDUS with normal or elevated uptake. PCIV test with iodide discharge above 20% from basal rates was considered positive. Otorhinolaryngologist evaluation and tonal audiometric exam rule out sensorineural deafness. I organification defect was established in patients with eutopic thyroid, high serum TG, increased 131I uptake, positive PCIV test and normal hearing. TPO exons and exon-intron boundaries were amplified by PCR and automatic sequenced. Results: From 40 patients with CH, seven patients (3F:4M) had the diagnosis of I organification. Chronological age at diagnosis ranged from 3 to 10 years-old. Twelve polymorphisms previously described were detected. TPO mutations were identified in 3 non-consanguineous patients. A novel mutation R584Q was found in homozygous state and in compound heterozygous state with Q660E mutation. Q660E was also found in heterozygous state but none mutation was found in the other allele. R584Q was not found in 50 normal controls. Patients with identified TPO mutation in both alleles showed the highest PCIV test, compatible to severity of defects. Q660E had been previously described also in heterozygous state (2), suggesting severe TPO defect or an occult mutation in promoter region. Although I organification defect was suspected, TPO mutation was not found in the remaining patients. Therefore other gene must be studied to clarify the genetic diagnosis.[br]Conclusion: TPO gene mutations were found in 43% patients with suggestive I organification defects.[br][br](1) Beltrao CB et al., Endocrine J 2010;57:587-93. (2) Rodrigues C et al., Eur J Endocrinol. 2005; 152:193-8.[br][br]Sources of Research Support: FAPESP 2011/02627-5 and 2012/00324-8.[br][br]Nothing to Disclose: ESB, CBB, SM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 848 62 477 SAT-388 PO38-01 Saturday 416 2012


417 ENDO12L_SAT-389 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Hypophysiotropic TRH Neurons Are Stimulated by an Acute Increase in Physical Activity, While Inhibited by Two Weeks of Exercise (Wheel Running) Rosa M Uribe, Mariana Gutierrez-Mariscal, Candy Ramirez-Martinez, Edith Sanchez, Jean-Louis Charli, Patricia Joseph-Bravo Universidad Nacional Autonoma de Mexico, Cuernavaca, Mexico; Instituto Nacional de Psiquiatr[iacute]a Ram[oacute]n de la Fuente Mu[ntilde]iz, Mexico City, Mexico Hypothalamus-pituitary-thyroid (HPT) axis activity, essential for energy homeostasis, is inhibited by chronic stress but effects of acute stress are contradictory. We hypothesized that a determinant component is the energy demand of the physical activity imposed by some tests. Endocrine response was evaluated, within 1h of test termination, in Wistar male rats exposed 5 min to the elevated plus maze (EPM), or the open field test (OFT) during light (L) or dark (D) phases, or to 30 min restraint. All paradigms increased serum corticosterone concentration, and the levels of either CRH-R1 or proCRH mRNAs in the PVN. The HPT axis was activated after EPM-L and OFT-D (increased levels of PVN-proTRH mRNA, serum TSH), while inhibited after restraint (decreased levels of proTRH mRNA and serum TSH, increased TRH content in mediobasal hypothalamus (MBH) implying reduced release) or OFT-L (increased MBH content), which caused the highest corticosterone release. Changes in PVN proTRH expression or MBH TRH release, and serum TSH levels correlated positively with locomotion, while negatively with serum corticosterone values. PVN TRH neurons responsive to OFT-D were localized by in situ hybridization (ISH) to the middle PVN, where most TRH hypophysiotropic neurons reside. To evaluate the effects of exercise, rats were exposed to voluntary wheel running over-night for 1-14 days. Serum corticosterone levels increased on days 1 and 3, when body weight gain halted; body weight increased thereafter less intensely than in control rats. Weight of brown and white adipose tissues decreased since day 3 stabilizing at a lower value than in controls by days 7-14. Levels of PVN proTRH mRNA or MBH-TRH were unchanged, those of serum TSH gradually decreased since the first day, those of serum T4 had a tendency to decrease by day 14, while those of serum T3, significantly on day 1 only. Variations in T3 or T4 correlated positively with travelled distance on days 3 and 7; those in proTRH mRNA correlated positively with travelled distance on day 3, while negatively with body weight by day 14. ISH revealed a small but significant decrease in proTRH expression in middle-PVN by day 14. Thus, the activity of HPT axis depends on a balance between the stimulatory effect of acute increased physical activity and the negative impact of stress-induced enhancement of serum corticosterone levels; however, the long-term negative energy balance imposed by training, resets the axis at a lower level.[br][br]Sources of Research Support: CONACYT 83363.[br][br]Nothing to Disclose: RMU, MG-M, CR-M, ES, J-LC, PJ-B 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1294 62 478 SAT-389 PO38-01 Saturday 417 2012


418 ENDO12L_SAT-390 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) DNA Methylation as a Predisposing Factor in the Pathogenesis of Congenital Hypothyroidism in Twin Births Federica Marelli, Giovanna Weber, Maria Cristina Vigone, Giorgio Radetti, Davide Gentilini, Luca Persani Universit[agrave] degli Studi di Milano, Milan, Italy; San Raffaele Hospital, Milan, Italy; Hospital of Bolzan, Bolzan, Italy; IRCCS Istituto Auxologico Italiano, Cusano Milanino, Italy Several reports indicate a 3-fold higher incidence of congenital hypothyroidism (CH) in multiple as compared to single gestations, but the underlying explanations of such phenomenon are still unknown. The investigation of discordant and concordant twin pairs is a powerful strategy for uncovering disease-associated epigenetic changes because it allows an assessment of the epigenome independent of any underlying genomic sequence variation. This project is focused on the study of DNA methylation, as predisposing factor of thyroid diseases in twin births. We collected a total of six twin pairs discordant for CH (DTP: 2 monozygotic, MZ, and 4 dizygotic pairs, DZ) and other six twin pairs concordant for CH (CTP: 4 MZ and 2 DZ pairs) and analyzed the global DNA methylation levels by the Illumina Infinium HumanMethylation27 BeadChip. The unsupervised hierarchal clustering of these subjects based on their methylation profile showed that birth weight (BW) appears the main factor associated with differences of the epigenome. In fact, DTP and CTP with similar BW, regardless of their gestational ages (GA) clustered together, in contrast with subjects with different BWs. Interestingly, within the DTP group, all of the CH-twins have a lower BW than euthyroid twins, suggesting a possible relationship between DNA methylation, low BW and thyroid defects. The global methylation analysis revealed that CH affected twins with low BW are significantly hypomethylated than hypothyroid or euthyroid counterparts. In particular, the 60-70% of differentially methylated CpG sites are located in DNA non-coding regions suggesting the existence of a genomic instability effect correlated with low BW. Since maternal folate deficiency caused by MTHFR deficiency leads to a shortage of methyl-donors and to a consequent hypomethylation status, we analyzed the presence of MTHFR variants in the maternal DNA of these differentially methylated twin pairs. Interestingly, we found that 2 mothers are homozygous and one is heterozygous for the MTHFR 677C/T variant, and 2 mothers have the MTHFR 1298A/C genotype. The presence of these genetic variants was invariably associated with low BW and CH and may therefore potentially account for the fetal hypomethylated status.[br]In conclusion in this group of twin gestations, CH appears associated with low birth weight, rather than gestational age, as well as with a global DNA methylation defect as a possible consequence of maternal MTHFR variations.[br][br]Nothing to Disclose: FM, GW, MCV, GR, DG, LP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1561 62 479 SAT-390 PO38-01 Saturday 418 2012


419 ENDO12L_SAT-391 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Lipopolysaccharide (LPS)-Induced Non-Thyroidal Illness Syndrome (NTIS) in Pigs Causes Profound Hypothyroidism and Tissue-Specific Changes in Thyroid Hormone Metabolism and Molecular Targets Isabel Castro-Piedras, Leah Quisenberry, James Hutson, Reid Norman, John McGlone, Daniel Hardy, Joaquin Lado-Abeal Texas Technical University Health Sciences Center, Lubbock, TX; Texas Technical University Health Sciences Center, Lubbock, TX; Texas Technical University Health Sciences Center, Lubbock, TX; Texas Technical University Health Sciences Center, Lubbock, TX [bold]Introduction. [/bold]The effect of sepsis on thyroid hormone (TH) levels and TH molecular targets, as well as the relationship between NF-kB activation and THRB gene expression was investigated in a large animal model of septic shock NTIS.[br][bold]Material and Methods. [/bold]17 domestic pigs (9-11 kg) were given saline (control, n=8) or LPS (n=9): 3.5 [mu]g/kg/min for 5 minutes followed by 3.5 [mu]g/kg/hr intravenously for 48 hr. Tissue samples were obtained from multiple organs after euthanasia. Tissue samples were assayed for total T4 and T3, and serum samples were assayed for free T4 (FT4), FT3, reverse T3 (rT3) and cortisol. Expression of MCT8, THRB, THRA1, THRA2, RXRA, RXRB, NCOR1, DIO1, DIO2, and, RPL4 (internal control) genes were assessed by qPCR. D1, D2 and D3 activity were assayed in tissue homogenates. NF-kB activation was evaluated by EMSA on nuclear extracts incubated with [sup]32[/sup]P-oligonucleotides containing a NF-kB binding site.[br][bold]Results. [/bold]Pigs given LPS had lower serum and tissue TH levels than control pigs. D1 activity was detected only in liver and kidney (tissues with the highest DIO1 expression). Endotoxemia decreased DIO1 expression in both liver and kidney. DIO2 expression and D2 activity were detected in all studied tissues. DIO2 expression increased in heart, kidney and spleen of LPS treated pigs compared to control pigs, while no significant differences were observed in D2 activity. D3 activity was detected in all tissues, and endotoxemia was related to increased D3 activity in hypothalamus, thyroid, heart and liver. The other studied genes showed tissue specific changes in expression. In frontal lobe, cerebellum, hypothalamus, thyroid, and skeletal muscle, no significant differences were observed between groups; however, LPS caused a profound decrease in MCT8 and THRB expression in heart, aorta, liver, kidney, and adrenal gland. Also, THRA1 decreased in heart, adrenal gland and kidney medulla of septic animals. No relationship between changes in THRB expression and NF-kB activation was observed.[br][bold]Conclusions. [/bold]LPS treated pigs showed changes in serum TH levels that resemble those in humans with septic shock NTIS. A profound hypothyroidism was observed in tissues from animals with endotoxemia. The changes in D1 and D3 activities during sepsis contributed to NTIS hypothyroidism. THRB, THRA1 and MCT8 gene repression during sepsis was tissue specific, and TRHB repression did not correlate with NF-kB pathway activation.[br][br]Sources of Research Support: CH Foundation. Spanish Ministry of Education (MEC).[br][br]Nothing to Disclose: IC-P, LQ, JH, RN, JM, DH, JL-A 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1653 62 480 SAT-391 PO38-01 Saturday 419 2012


420 ENDO12L_SAT-392 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Fasting Induces a Delayed Activation of Thyrotropin-Releasing Hormone Degrading Ectoenzyme in Tanycytes of the Mediobasal Hypothalamus of Adult Rats Ivan Lazcano-Sanchez, Jean-Louis Charli, Patricia Joseph-Bravo, Miguel Cisneros, Gabriel Linares, Edith Sanchez Instituto Nacional de Psiquiatr[iacute]a Ram[oacute]n de la Fuente Mu[ntilde]iz, M[eacute]xico City, Mexico; Instituto de Biotecnolog[iacute]a, Universidad Nacional Aut[oacute]noma de M[eacute]xico, Cuernavaca, Mexico Fasting induces a rapid reduction of the activity of the hypothalamic- pituitary-thyroid (HPT) axis, with a drop in the synthesis of thyrotropin releasing hormone (TRH) in hypophysiotropic neurons of the paraventricular nucleus (PVN) and decreased TRH release into the pituitary portal capillaries. As pyroglutamyl peptidase II (PPII), the TRH degrading ectoenzyme, is expressed by the [beta]2 tanycytes, in close apposition to the TRH nerve endings in the external layer of the median eminence (1), we tested the hypothesis that tanycyte PPII activity is regulated during fasting. PPII mRNA expression and activity was measured in the median eminence of male Wistar rats (225-300 g BW) submitted to fasting for 24-72 h, with water ad libitum. Since the role of thyroliberinase, the isoform of PPII that is secreted by the liver into the circulation, is poorly understood, we also measured thyroliberinase activity in serum. Control rats were submitted to isolation for the same period of time than food-deprived animals and sacrificed between 3-5 h after lights were on. Compared to isolation, fasting reduced significantly animal weight since 36 h [Fed ad lib: 100%[plusmn]1; fasted 90%[plusmn]1; P[lt]0.001], PVN TRH mRNA levels at 48 h, measured by in situ hybridization histochemistry on coronal sections of the hypothalamus, tended to reduce serum TSH and total T3 concentrations between 36-60 h, and decreased total concentration of T4 in serum since 48 h [Fed ad lib: 65[plusmn]7; fasted 39[plusmn]4 nmol/L; P[lt]0.01]. Semi-quantitative in situ hybridization indicated that PPII mRNA levels increased in [beta]2 tanycytes 48 h after fasting initiation. This increase was transitory (not detected at 72 h), and followed by an increase of PPII activity in the median eminence at 72 h [Fed ad lib: 6.5[plusmn]1.3; fasted 12.5[plusmn]0.7 pmoles [beta]NA/min/mg prot; P[lt]0.01]. Thyroliberinase activity increased since 36 h, with a peak at 48 h [Fed ad lib: 27[plusmn]5; fasted 44[plusmn]7 pmoles [beta]NA/min/ml; P[lt]0.05]. We conclude that during fasting the delayed increase in median eminence PPII activity, as well as that of thyroliberinase, may facilitate the maintenance of the profound reduction of HPT axis activity.[br][br](1) S[aacute]nchez E et al., Endocrinology 2009; 150:2283.[br][br]Sources of Research Support: CONACYT 107109 (ES), 61804 (JLC) y 83363 (PJB).[br][br]Nothing to Disclose: IL-S, J-LC, PJ-B, MC, GL, ES 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1340 62 481 SAT-392 PO38-01 Saturday 420 2012


421 ENDO12L_SAT-393 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Effect of Thyroid Hormones in Hexokinase Activity and Subcellular Distribution in Rodents Tissues Flavia Leticia Martins Pecanha, Reinaldo Sousa dos Santos, Wagner Seixas da Silva Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil Thyroid hormones, T3 and T4, are very important in organism metabolism, regulating energy expenditure. Hexokinase (HK) is an important enzyme in glucose consume, responsible for the first step of glycolysis, catalyzing the conversion of glucose into glucose 6-phosphate. Our results, in mice, showed an increased activity of mitochondrial HK (mitHK) in cardiac, solear and gastrocnemius muscle of hyperthyroid groups. The objective of our work is to investigate if the same occurs in rats and try to identify possible mechanism related to this phenomenon. To achieve our objectives we used two different models, rat and C2C12 cell line. Wistar rats groups were treated as describe below: hypothyroid [ndash] received 0,03 % (w/v) of methimazole in drink water during 21 days; hyperthyroid [ndash] received 0,1 [mu]g T4/g body mass during 10 days; euthyroid [ndash] received both vehicles. Cells were cultivated in stripped medium supplemented with 150 pM, 740 pM and 14800 pM T3 to achieve hypo-, eu- and hyperthyroid status. In rats, we observed an increase in heart/body mass ratio. We also observed an increase in mitHK activity of solear and heart, as described in mice. In gastrocnemius muscle we did not see mitHK activity. In an attempt to investigate the mechanisms of hexokinase modulation, we observed that T3 had no direct effect on HK activity. Furthermore, the expression of mitHK did not change after treatment in all groups. In C2C12, a muscle cell line, as well as other muscle tissues, we observed an increase in mitHK activity from hyperthyroid group after 24h of treatment. Moreover, we noted a decreased in mitHK activity from hypothyroid group. We did not see any change in soluble HK activity among the groups. We also noticed that increased HK activity appeared between 5 and 25 hours. This result together with one that shows that HK expression did not change, suggest that probably there is a factor that is regulated pre transcriptionally and then are able to modulate HK activity.[br][br]Sources of Research Support: Conselho Nacional de Desenvolvimento Cientifico e Tecnologico/Brazil, Fundacao Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro/Brazil.[br][br]Nothing to Disclose: FLMP, RSdS, WSdS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 630 62 482 SAT-393 PO38-01 Saturday 421 2012


422 ENDO12L_SAT-394 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Effect of Thyroid Hormone to the Expression of Bile Salt Export Pump and ATP-Binding Cassette Transporter Hwa Young Ahn, Jung Ah Lim, Hoon Sung Choi, Jae Hoon Moon, Yun Jae Chung, Young Joo Park, Ka Hee Yi, Do Joon Park, Seong Yeon Kim Seoul National Univesity College of Medicine, Seoul, Korea; Seoul National University Bundang Hospital, Seongnam, Korea; Chung-Ang University College of Medicine, Seoul, Korea Introduction[bold]: [/bold]Excretion of bile salt and free cholesterol through bile was essential for intestinal digestion, absorption of lipids and regulation of cholesterol. Thyroid hormone plays an important role in the production of bile acids through regulation of CYP7A1 (cholesterol 7[alpha]-hydroxylase) expression. However, it is not well known whether thyroid hormone contributes excretion of bile acids.[br]In this study, we aimed to evaluate the effect of thyroid hormone to expression of bile salt export pump (BSEP) and ATP-binding cassette transporter, ABCG5 and ABCG8 in the liver.[br]Material and Methods: We injected thyroid hormone (T3) to C57BL/6 mice intraperitoneally and then RNA and protein were isolated from liver. After 6 hours and 5 days of T3 treatment, we measured serum and hepatic cholesterol and hepatobiliary bile acids.T3 was administered to mouse primary hepatocytes and human hetatoma cell to evaluate in vitro effect.[br]Results: Serum total cholesterol was decreased after 5 days of T3 treatment. Expression of BSEP mRNA and protein in mouse liver were increased after T3 treatment. BSEP protein in human hepatoma cell was also increased after T3 treatment. Expression of ABCG5/8 mRNA and ABCG5 protein were increased after T3 treatment. In primary hepatocytes, T3 also increased ABCG5/8 mRNA expression. Nuclear receptor, liver X receptor (LXR) has been suggested to regulate the expression of ABCG5/8. When we cotransfected liver X receptor response element (LXRE) construct and thyroid hormone receptor (TR)[beta]/retinoid X receptor (RXR)[alpha] with T3, the activity of LXRE containing construct was markedly increased.[br]Conclusion: We confirmed that thyroid hormone increased expression of BSEP and ABCG5/8. These results suggested that thyroid hormone can regulate cholesterol metabolism, not only increased bile acid synthesis but also stimulated the excretion of bile acids through increased expression of BSEP and ABCG5/8.[br][br]Nothing to Disclose: HYA, JAL, HSC, JHM, YJC, YJP, KHY, DJP, SYK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 502 62 483 SAT-394 PO38-01 Saturday 422 2012


423 ENDO12L_SAT-395 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Thyroid Hormone and Estrogen Overlapping Actions in Kidney Glutathione S-Transferase Alpha Gene Expression Regulation Larissa C Faustino, Norma A Almeida, Guilherme F Pereira, Marcelo M Morales, Carmen C Pazos-Moura, Tania M Ortiga-Carvalho Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil; Universidade Federal Rural do Rio de Janeiro, Serop[eacute]dica, Brazil Glutathione S-transferases (GST) are a superfamily of ubiquitous dimeric detoxification isoenzymes that conjugate many substrates to reduced glutathione. Alpha class GST (Gsta) is an essential component of antioxidant defence mechanism both in liver and kidney. Hypothyroidism up regulates Gsta expression in liver of male or female mice (Faustino, 2011) however there is no published information regarding kidney Gsta regulation by thyroid hormones or sex steroids. Our previous data regarding the kidney showed that in males, there is an increase of Gsta expression in hypothyroidism. Surprisingly, in females there is a reduction of renal Gsta levels. Here, we investigated, if female renal Gsta levels are a result of a cross-talk among T3 and estrogen. Mice were subjected to ovariectomy (OVX) and then, induced to hypothyroidism (HYPO - PTU diet for 4 weeks) and hyperthyroidism (HYPER, T3-50[mu]g/100g BW/15days). In a second experiment, after OVX, females were treated with low (0.01mg) or high (0.1mg/kg BW) dose of estradiol benzoate (Es). In a final experiment, after OVX and TH modulation, mice were treated with estradiol benzoate (0.1mg/kg BW). Protein and mRNA levels were measured by real-time RT-PCR and Western Blotting. By kidney tubular micro-dissection, we showed that both males and females expressed Gsta mostly in the proximal tubule and that females had increased mRNA levels of Gsta compared to males (80% more). After ovariectomy (OVX), Gsta mRNA was elevated in HYPO (2 times) compared to EU and HYPER. Protein levels showed the same results as the mRNA. OVX mice plus low and high doses of Es presented two-times of mRNA levels compared to OVX alone and males (0.94[plusmn]0.2). We next evaluated TR[beta]1 and ER[alpha] mRNA renal levels. OVX mice exhibited higher mRNA levels of TR[beta]1 (1.0[plusmn]0.05) and ER[alpha] (1.0[plusmn]0.02) than sham (TR[beta]1:0.6[plusmn]0.1*; ER[alpha]: 0.4[plusmn]0.06*) and OVX plus ES mice (TR[beta]1:0.4[plusmn]0.06*; ER[alpha]: 0.3[plusmn]0.03*), suggesting down regulation of both receptors by estrogen. Finally, we analyzed Gsta expression in OVX mice, plus Es, at different thyroid states. Interestingly, the expression pattern was opposite to that found in OVX alone in EU (ovx: 1.0[plusmn]0.05; ovx +Es: 1.7[plusmn]0.1*), HYPO (ovx: 2.1[plusmn]0.2; ovx+Es: 1.2[plusmn] 0.15*) and HYPER (ovx: 1.5[plusmn]0.4; ovx+Es: 2.9[plusmn]0.4*). Our data show overlapping actions of T3 and estradiol regulating renal Gsta expression, in which Gsta is down regulated by T3 in the absence of estradiol, and up regulated by T3 when estradiol is present. * P[lt]0.05 or less.[br][br]Faustino et al., Exp Physiol 2011; 96:790.[br][br]Sources of Research Support: CNPq, CAPES, FAPERJ, Dept. of Thyroid, SBEM.[br][br]Nothing to Disclose: LCF, NAA, GFP, MMM, CCP-M, TMO-C 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 640 62 484 SAT-395 PO38-01 Saturday 423 2012


424 ENDO12L_SAT-396 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Regulation of Bile Acid Synthesis in Humans by Thyroid Hormone Jan Lammel, Stephen Ayers, Anusha Angajala, Paul Webb Methodist Hospital Research Institute, Houston, TX Thyroid hormone is known to regulate hepatic cholesterol metabolism through the transcriptional regulation of metabolic gene sets through its actions on thyroid hormone receptor beta (THRB) in liver. Although this regulation has been observed to include the induction of bile acid synthesis in various animal models as well as in humans, studies of the mechanism of this effect have produced inconsistent results in different model systems. In mouse models, this conversion has been explained through the direct transcriptional regulation of Cyp7a1. In this investigation, we have performed a further characterization of thyroid hormone[apos]s regulation of hepatic bile acid synthesis in mouse and human hepatoctye cell models. Through this analysis, we have observed the regulation of bile acid metabolic genes, including Cyp7a1, leading to the characterization of distinct proximal regulatory elements, and providing an explanation for the regulation of this metabolic process. Our observations provide a consistent explanation for previous observations in human patients and animal models, and add insight to the mechanism for the regulation of serum lipid parameters by thyroid hormone.[br][br]Nothing to Disclose: JL, SA, AA, PW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2317 62 485 SAT-396 PO38-01 Saturday 424 2012


425 ENDO12L_SAT-397 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Thyroid Hormone Action on Cerebrocortical Neurons in Primary Culture Pilar Gil-Ibanez, Juan Bernal, Beatriz Morte Instituto de Investigaciones Biom[eacute]dicas [quot]Alberto Sols[quot], Madrid, Spain; CIBERER, Madrid, Spain Thyroid hormone (T3) is important during development of the mammalian brain acting through regulation of gene expression. T3 is involved in neuronal and glial cell differentiation and migration, axonal myelination, and synaptogenesis. To get a further insight on the genes and pathways regulated by T3 in neural cells, we used mouse cerebrocortical neurons in primary culture. To identify thyroid hormone-dependent genes we used microarrays. A bioinformatics analysis of these genes predicted a crosstalk between thyroid hormone signaling and glucocorticoid and retinoid pathways. We confirmed synergistic gene regulation between T3 and dexamethasone. We also demonstrated that T3 regulates the expression of enzymes of the retinoic acid metabolism thereby influencing the retinoid pathway. In this study we have identified many genes regulated at the cellular level by thyroid hormone in neurons. The interrelation of T3, retinoid and glucocorticoid signaling in the regulation of neuronal gene expression should be relevant on stage-specific processes of brain development.[br][br]Nothing to Disclose: PG-I, JB, BM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 753 62 486 SAT-397 PO38-01 Saturday 425 2012


426 ENDO12L_SAT-398 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Thyroid Hormone Receptor [beta] Is a Suppressor of SV40-Mediated Tumorigenesis Dong Wook Kim, Li Zhao, Sheue-Yann Cheng National Cancer Institute, National Institutes of Health, Bethesda, MD Accumulated evidence suggests that thyroid hormone receptor [beta] (TR[beta]) could suppress tumor cell proliferation and invasion, but the detailed mechanisms by which TR[beta] inhibits tumor development is not fully understood. Using simian virus-40 (SV40)-induced tumorigenesis, we explored the mechanisms by which TR[beta] acts to inhibit tumor development and growth. SV40 large T antigen (Tag)-immortalized human thyroid epithelial (HTori) cells stably expressing TR[beta] (HTori-TR[beta]) or only the selectable neomycin gene (HTori-Neo) were injected into immune-deficient mice. HTori-Neo, but not HTori-TR[beta], cells induced tumors, indicating TR[beta] acted to suppress SV40-Tag-induced tumor development. The SV40-Tag oncoprotein binds to and inactivates the retinoblastoma protein (Rb) and the tumor suppressor p53, thereby inducing tumorigenesis. We hypothesized that TR[beta] could suppress tumorigenesis by interfering with SV40-Tag binding to Rb and p53. Indeed, cell-based studies showed that TR[beta] competed with Rb for binding to SV40-Tag oncoprotein. Rb, released from SV40-Tag oncoprotein, altered signaling pathways to impede cell cycle progression. The protein abundance of cyclin E, a key regulator in the G1/S progression, was lower in HTori-TR[beta] cells than in HTori-Neo cells with concomitant reduced phosphorylated Rb(S807/811). These changes were accompanied by reduced cell proliferation and markedly delayed cell entry from the G1 to the S phase, indicating a TR[beta]-mediated inhibition of cell proliferation. TR[beta] also competed with p53 for binding to SV40-Tag, as evidenced by decreased association of p53 with SV40-Tag in HTori-TR[beta] cells. PTEN, a direct p53 target gene, was found to increase expression at the mRNA and protein levels in HTori-TR[beta] cells, leading to decreased phosphorylated Akt(S473) to attenuate downstream signaling, thereby decreasing cell proliferation. Moreover, the protein abundance of Bcl2, an anti-apoptotic regulator that is a downstream effector of p53, was decreased in HTori-TR[beta] cells, accompanied by increased protein abundance of BAX, an apoptosis activator. These alterations led to increased apoptosis, as evidenced by elevated expression of cleaved caspase-3 and of cleaved PARP in HTori-TR[beta] cells. Our results show that TR[beta] suppressed tumorigenesis by decreasing cell proliferation and increasing apoptosis. Our study uncovered a novel mechanism by which TR[beta] inhibits tumorigenesis by blocking the oncogenic actions of SV40-Tag via protein-protein interaction.[br][br]Nothing to Disclose: DK, LZ, S-YC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 424 62 487 SAT-398 PO38-01 Saturday 426 2012


427 ENDO12L_SAT-399 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Tetraiodothyroacetic Acid (Tetrac) Acts at a Plasma Membrane Receptor To Modulate Expression of Inflammation-Related Genes in Cancer Cells Hung-Yun Lin, Gennadi V Glinsky, Anna B Glinskii, Faith B Davis, Shaker A Mousa, Mary K Luidens, Aleck Hercbergs, Paul J Davis Albany College of Pharmacy and Health Sciences, Albany, NY; Signal Transduction, Inc, Albany, NY; Albany Medical College, Albany, NY; Cleveland Clinic, Cleveland, OH The inflammatory response has contributions from inflammatory cells, response-modifying proteins (cytokines) and blood vessel growth fractors. Agonist thyroid hormones (L-thyroxine, T4; 3, 5, 3[apos]-triiodo-L-thyronine, T3) act at a cell surface receptor on integrin [alpha]v[beta]3 to stimulate white blood cell mirgation, actions of cytokines, e.g., interferon-[gamma] on HLA-DR expression, and angiogenesis, the latter via four vascular growth factors (1). These actions onf T4 and T3 are blocked at the integrin by tetraiodothyroacetic acid (tetrac) and its nanoparticulate formulation (nanotetrac); the latter acts exclusively at the [alpha]v[beta]3 hormone receptor and does not enter the cell. The interaction of inflammation and tumor cell biology is relevant to cancers of several organs, including breast, liver and gastrointestinal tract. We have previously reported that tetrac and nanotetrac, via mechanisms initiated at the cell surface, modulate the expression of a number of cancer cell survival pathways genes (2). We describe here the actions of tetrac (10[sup]-7[/sup] M) or nanotetrac (10[sup]-7[/sup] M tetrac equivalent at 24 h on expression of genes in human breast cancer (MDA-MB-231) cells that are relevant to inflammation. Nanotetrac significantly (P [lt]0.05) suppressed expression of 5 of 6 differentially-regulated interleukin genes, including [italic]IL-6[/italic] and [italic]IL-1[alpha][/italic], but stimulated expression of [italic]IL-11[/italic], a factor that supports proliferation of hematopoietic stem cells and megakaryocyte progenitor cells. Nanotetrac reduced expression of most interferon response pathway genes, e.g., [italic]IRF-5[/italic] and [italic]IFIT2[/italic], but had a minimally suppressive effect on [italic]IRF1[/italic]. Tetrac and nanotetrac each stimulated expression of [italic]SOCS4[/italic], whose gene product is an endogenous suppressor of cytokine signaling. Nanotetrac downregulated expression of chemokine and chemokine receptor genes, including [italic]CX3CL1[/italic] ([italic]fractalkine[/italic]) and its receptor, [italic]CX3CR1[/italic]. Thus, nanotetrac has coherent effects on expression of several families of genes whose products contribute importantly to inflammation; the effects are anti-inflammatory in pattern and are desirable from the standpoint of potentially minimizing contributions of inflammation to the support or emergence of inflammation-associated cancers. Certain of the cytokine genes affected by nanotetrac are also important to atherogenesis, warranting analysis of anti-inflammatory activity of nanotetrac in blood vessel cells.[br][br](1) Davis PJ et al., Annu Rev Pharmacol Toxicol 2011; 51:99. (2) Glinskii AB et al., Cell Cycle 2009; 8:3562.[br][br]Nothing to Disclose: H-YL, GVG, ABG, FBD, SAM, MKL, AH, PJD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2367 62 488 SAT-399 PO38-01 Saturday 427 2012


428 ENDO12L_SAT-400 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Assessment of Thyroid Status Using Monocyte Gene Expression J Jonklaas, J Credle, M Danielsen Georgetown University, Washington, DC; Georgetown University, Washington, DC Measurement of gene expression in tissues such as liver and muscle is not routinely used for clinical assessment of thyroid status, as it requires tissue biopsy. Peripheral blood samples, however, can be obtained with minimal discomfort and used to measure genes expressed in human blood cells. To develop a gene expression system that reflects thyroid status, we have used microarray analysis to identify genes in peripheral monocytes that have mRNA expression levels that respond to thyroid hormone. We chose circulating monocytes because they are easily purified from blood and reportedly have a transcriptional response to TH.[br]We used the human Affymetrix U133a gene array to compare gene expression in monocytes from patients with hypothyroidism to gene expression in the same patient after euthyroidism was achieved. We were able to separate the hypothyroid and euthyroid groups by unsupervised hierarchical clustering of differentially expressed genes, showing that gene expression patterns of the two groups are indeed unique. From these studies we identified 38 genes that have at least a 2-fold change in expression levels.[br]In our in vivo study, hypothyroid individuals had high TSH levels that then decreased after TH treatment. The effects we observed on gene expression changes could therefore be due not only to changes in TH levels but to changes in TSH levels. To show that TH directly controls gene expression in monocytes, we measured gene expression of a subset of genes induced by TH in vivo using TH treated monocytes ex vivo. We examined 8 genes and found that their expression levels did indeed increase upon the addition of T3 to the culture medium. Our data also show that these monocytes express both thyroid hormone receptors alpha and beta, and that mRNA levels for both receptors are induced by T3.[br]In summary, the expression of certain monocyte genes can serve as a tissue biomarker of thyroid status. This biomarker, in addition to providing assessment of thyroid status in patients with TSH deficiency, could provide a means of assessing tissue euthyroidism during therapy with LT4 and during combination therapy with LT4 and T3.[br][br]Sources of Research Support: Funding provided by the Georgetown Howard Universities Clinical and Translational Science Award (1UL1RR031975), and a Translational Science Award from Georgetown University Department of Medicine.[br][br]Nothing to Disclose: JJ, JC, MD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 326 62 489 SAT-400 PO38-01 Saturday 428 2012


429 ENDO12L_SAT-401 POSTER SESSION: The Hypothalamic-Pituitary-Thyroid Axis [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Effect of Neonatal Nicotine Exposure on Thyroid Hormone Metabolism in Rat Patricia C Lisboa, Elaine Oliveira, Tania M Ortiga-Carvalho, Isis H Trevenzoli, Ana P Santos-Silva, Cintia R Pinheiro, Larissa C Faustino, Alex C Manhaes, Egberto G Moura State University of Rio de Janeiro, Rio de Janeiro, Brazil; Federal University of Rio de Janeiro, Rio de Janeiro, Brazil Background: Postnatal nicotine exposure causes early primary hypothyroidism and programs for secondary hypothyroidism at adulthood (1,2). Objectives: We studied the effects of maternal nicotine exposure during lactation on the thyroid hormone (TH) metabolism of neonate and adult offspring. Methods: Osmotic minipumps were implanted in lactating rats, and nicotine (NIC, 6 mg/kg/day s.c.) or saline (control) was released from day 2 to 16. NIC and control offspring were killed at 15 and 180 days-old (n=10 per group, one rat per litter). We measured type 1 and 2 deiodinase activity and expression, and mitochondrial [alpha]-glycerol-3-phosphate dehydrogenase (mGPD) activity. Significant differences had p[lt]0.05. Results: NIC exposure caused lower liver 5[apos]-D1 (-26%) and mGPD (-29%) activities at 15 as well as at 180 days-old (5[apos]-D1:-57%; mGPD:-41%) that are in accordance with offspring[apos]s thyroid status on both periods. Adult NIC offspring showed lower muscle 5[apos]-D1 activity (-47%), higher 5[apos]-D2 activity in brown adipose tissue (BAT, 5.2-fold increase), heart (+35%) and testis (+39%). The expression of deiodinase mRNAs in liver, muscle, BAT, heart and testis followed the same profile of the enzymatic activity. As expected, those TH-dependent enzymes are in accordance with hypothyroidism. However, deiodinase activities were not changed in hypothalamus, pituitary and thyroid of adult NIC offspring, suggesting that homeostatic mechanisms preserve TH metabolism in these tissues. Conclusions: For the first time, short and long-term consequences of postnatal nicotine exposure on TH peripheral conversion in the progeny were demonstrated.[br][br](1) Oliveira E et al. J Endocrinol 2009; 202: 397. (2) Santos-Silva AP et al. J Endocrinol 2010; 209:1.[br][br]Sources of Research Support: CNPq, Capes and Faperj.[br][br]Nothing to Disclose: PCL, EO, TMO-C, IHT, APS-S, CRP, LCF, ACM, EGM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1440 62 490 SAT-401 PO38-01 Saturday 429 2012


430 ENDO12L_SAT-402 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) A Solitary Hyperfunctioning Thyroid Nodule Harboring Follicular Thyroid Carcinoma: Case Report and Review of the Literature Sasan Mirfakhraee, Dana Mathews, Lan Peng, Stacey L Woodruff, Jeffrey M Zigman University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX [bold] Background: [/bold] Classically, thyroid nodules that are autonomously hyperfunctioning on scintigraphy represent benign adenomas. However, there are a number of case reports describing hyperfunctioning thyroid nodules that were determined to be thyroid carcinoma following resection. We present the case of a patient with a [ldquo]hot[rdquo] nodule found to be follicular thyroid carcinoma on surgical pathology and then review similarly-described cases in the literature.[br][bold] Clinical Case: [/bold] A 29-year old woman presented with a thyroid nodule. She denied prior radiation exposure or family history of thyroid cancer. She endorsed mild thyrotoxic symptoms. Her TSH was suppressed (0.005mcIU/mL, normal 0.40[ndash]4.50mcIU/mL) but fT3 and fT4 were normal. Thyroid ultrasound showed a 2.7cm nodule that was solid and isoechoic without suspicious features. I-123 thyroid scan demonstrated that the nodule was autonomously hyperfunctioning with 24-hour iodine uptake of 27%. The patient underwent left hemithyroidectomy and pathologic examination revealed a solitary, well-encapsulated, minimally-invasive follicular carcinoma. The patient has remained euthyroid after surgery without evidence of cancer recurrence.[br][bold] Review of the Literature: [/bold] A literature search through January 2012 was performed using the PubMed/MEDLINE database; reference lists were explored to find additional cases. A total of 83 cases were found of autonomously hyperfunctioning thyroid nodules proven to be malignant on pathologic review. Of these nodules, 41% were follicular and 59% papillary thyroid carcinoma. The mean nodule size was 3.9 [plusmn]1.9cm. The average age of the subjects was 47 [plusmn]19 years; 72% of subjects were female. 63% of subjects had overt hyperthyroidism, 14% had subclinical hyperthyroidism, and 23% were euthyroid. Only 1 patient had a high-risk clinical history. Ultrasonography was infrequently obtained; suspicious features were noted in 6 of 23 patients undergoing ultrasound.[br][bold] Clinical Lesson: [/bold] While the vast majority of hyperfunctioning thyroid nodules are benign adenomas, a number of case reports are proven exceptions to this rule. Our literature review reveals that high risk history and clinical features are of limited utility in identifying malignant thyroid nodules. Further studies could help determine if malignant hyperfunctioning nodules have distinctive features that distinguish them from nonmalignant nodules. Also, a cost/benefit analysis of performing biopsies on hyperfunctioning nodules would be instructive.[br][br]Nothing to Disclose: SM, DM, LP, SLW, JMZ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1848 63 491 SAT-402 PO56-01 Saturday 430 2012


431 ENDO12L_SAT-403 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Incidentally Detected Medullary Thyroid Cancer in a Patient on Liraglutide [mdash] More Questions Than Answers! Subramanian Kannan, Deborah Chute, Robert Zimmerman Cleveland Clinic Foundation, Cleveland, OH; Cleveland Clinic Foundation, Cleveland, OH [bold]Background:[/bold] GLP-1 receptors are present in rodent C-cells and stimulates C-cell secretion of calcitonin. A statistically significant increase in malignant C-cell carcinoma was seen in male rats with high plasma liraglutide concentrations. In contrast, monkeys receiving higher doses of liraglutide for 20 months did not demonstrate increased calcitonin release or C-cell hyperplasia. We report a patient on Liraglutide with incidentally detected Medullary thyroid cancer (MTC) and discuss some of its pathophysiological and clinical implications.[br][bold]Case report:[/bold] A 62 yr postmenopausal female presented to the clinic for enrolling in weight loss programs. She had previously lost 30 lbs on a protein sparing modified fast but regained 50% of weight, a year before presentation. Her past history was significant for a large multinodular goiter. Biopsy of dominant nodules in past were consistent with benign histology. She also had Hypertension, Dyslipidemia and Primary Hypothyroidism on replacement dose of levothyroxine 75 mcg/d. She denied any exposure to radiation to her head and neck region in the past. There was no family history of cancers or hypercalcemia. On clinical exam she was an obese individual with BMI of 30 kg/m[sup]2[/sup]. She had a multinodular goiter of about 60 gms with no palpable lymphadenopathy. Her labs were remarkable for fasting blood glucose of 120 mg/dl, HbA1c of 6.4 and TSH of 0.4 IU/ml. The Patient was started on Liraglutide with dose titrated up to 1.8 mcg/day. She lost about 10 lbs in 4 months. At this point the patient preferred to have thyroidectomy for her thyroid nodules. She underwent total thyroidectomy and was incidentally detected to have a 3 mm focus of MTC and another focus of 1mm of papillary thyroid cancer (follicular variant). A MEN2/FMTC mutation analysis was negative. She is currently on 125 mcg of Thyroxine suppression with TSH of 0.24 IU/ml and at 6 month follow up calcitonin and CEA levels have been negative.[br]Discussion: Expression of GLP-1 receptors in normal human thyroid tissue has been controversial, but they are over-expressed in MTC. Whether incretin receptor stimulation plays a role in hyperplasia and neoplasms or only serves as a marker of genetic dysregulation in abnor-mal tissue is not known. Proper patient selection, clinical vigilance, measuring serum calcitonin and Thyroid Ultrasound in appropriate cases may be helpful in patients on Liraglutide.[br][br]Disclosures: RZ: Speaker, Novo Nordisk. Nothing to Disclose: SK, DC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1868 63 492 SAT-403 PO56-01 Saturday 431 2012


432 ENDO12L_SAT-404 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Thyroglobulin Elevation Due to Heterophilic Antibodies Interference in the Follow-Up of a Patient with Papillary Thyroid Carcinoma Andrea Kozak, Patricia Fainstein Day, Maria Fabiana Russo Picasso, Carmen Cabezon, Sergio Damilano, Cecilia Fenili Hospital Italiano de Buenos Aires, Caba, Argentina; Laboratorio de Bioanal[iacute]tica, Caba, Argentina Introduction: Thyroglobulin (Tg) levels are the most efficient markers of disease in the follow-up of differentiated thyroid carcinoma. Methodological interference by antithyroglobulin antibodies (ATGU) is widely known, but less common interference by [ldquo]hook-effect[rdquo] and heterophilic antibodies (HA) has also been reported. We report a case of methodological interference by HA in the measurement of Tg by a chemiluminescent immunometric assay (CLIA) in the follow-up of a patient with papillary thyroid carcinoma,and suggest a study protocol for these cases.[br]Case report: In 2002 a female 43 year-old patient underwent a near-total thyroidectomy for a unifocal 2 cm CPT. She underwent thyroid remnant ablation with 100 mCi of I[sup]131[/sup] and was started on an inhibitory dose of levothyroxine (T4). Twelve months after surgery T4 was withdrawn: the 5mCi- I[sup]131[/sup] whole body scan (WBS) was negative, TSH [gt] 70 uU/ml and Tg [lt] 2 ng/ml. ATG were always negative ([lt]0.9 UI/mL). As from 2004, Tg was progressively higher, with or without TSH stimulation, whilst several WBS were always negative. The patient underwent several imaging procedures to rule out recurrence of CPT, that were negative. After a [ldquo]shot in the dark[rdquo] 150 mCi dose of I[sup]131[/sup], WBS did not show any uptake. Tg under TSH inhibition reached 473 ng/ml. Meantime, the patient appeared in good health and routine lab tests were normal. The consistently high Tg levels in a clinically disease-free patient suggested methodological interferences in the determination of Tg. To investigate this further, several tests were performed on the patient[apos]s serum: 1) Serial dilutions of the serum did no show linear correlation. 2) Measurement of Tg and ATGU by an alternative assay (ECLIA) resulted in Tg[lt] 0.5 ng/ml and ATGU [lt]10 UI/ml. 3) Measurement of Tg by both assays after precipitation of protein A ruled out high molecular weight variants of Tg. 4)A recovery assay showed 100% recovery. 5) Treatment with a specific blocker of HA (HBT Scantibodies) and measurement of Tg by CLIA showed Tg[lt]0.9ng/ml, confirming methodological interference by HA.[br]Conclusions: We confirmed methodological interference by HA in the measurement of Tg in a patient with CPT by performing an in-depth study protocol. We therefore suggest using this protocol to identify methodological interference whenever Tg results are inconsistent with the clinical course of the patient and thereby avoid costly imaging studies and unnecessary treatments.[br][br]Nothing to Disclose: AK, PFD, MFRP, CC, SD, CF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 586 63 493 SAT-404 PO56-01 Saturday 432 2012


433 ENDO12L_SAT-405 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) A Case of Distant Metastatic Highly Differentiated Follicular Carcinoma of Ovarian Origin (HDFCO) and Concurrent Micropapillary Thyroid Carcinoma Brittany Noel Bohinc, Jennifer Marie Perkins Duke University Hospital, Durham, NC CASE: A 28-year-old female 6 weeks postpartum presented with pelvic pain and was found to have a 7.5 cm left ovarian mass. She underwent resection and pathology revealed struma ovarri with no malignant features. Nineteen years later, she presented with left-sided flank pain. A CT scan showed a 3.2 cm left periaortic lymph node. CT-guided biopsy revealed bland thyroid tissue with no malignant features. I-131 ablation was planned for treatment of metastatic highly-differentiated follicular carcinoma of ovarian origin (HDFCO). In preparation, a total thyroidectomy was performed. Pathology revealed a single 1 mm focus of papillary microcarcinoma. The patient underwent Thyrogen-stimulated I-131 ablation with 104.6 mCi I-131. Post-treatment whole body scan (WBS) revealed increased activity in the left periaortic region and another focus in the right supraclavicular region. Stimulated thyroglobulin (Tg) after treatment remained detectable at 26 ng/ml. Twelve months later, repeat WBS showed persistent foci of increased radiotracer activity in the right supraclavicular region and left mid-abdomen that appeared unchanged. She was retreated with 175 mCi I-131, continued on relatively low doses of levothyroxine for body weight because of persistently functioning ectopic thyroid tissue, and followed with serial imaging and Tg. Her most recent suppressed Tg was 0.3 ng/ml.[br]DISCUSSION: Metastatic struma ovarii is a rare diagnosis and is usually associated with areas of differentiated thyroid cancer (frequently papillary thyroid carcinoma) originating in the ovary (1-6). Recently, bland metastatic struma ovarii has also been described, coined as the term highly differentiated follicular carcinoma of the ovary (HDFCO)(7). This is a very rare diagnosis and is confusing to the treating physician with regards to prognosis and long-term follow-up as it continues to appear benign on pathology, but has metastatic potential. By definition, HDFCO tumors have an innocuous appearance resembling that of bland thyroid tissue in both the ovary and sites of dissemination. To our knowledge, this is the fifteenth known case with peritoneal spread and fifth case shown to metastasize to a distant site. We suspect the 1mm PTC found at thyroidectomy was incidental. The treatment of choice for patients with HDFCO is local resection of the extra-ovarian tumor (if possible), thyroidectomy, and radioactive iodine ablation (8).[br][br](1) Kabukcuoglu F, Baksu A, Yilmaz B, Aktumen A, Evren I 2002 Malignant struma ovarii. Pathol Oncol Res 8:145[ndash]147. (2) O[apos]Connell MEA, Fisher C, Harmer MB 1990 Malignant struma ovarii: presentation and management. Br J Radiol 63: 360[ndash]363. (3) Devaney K, Snyder R, Norris HJ, Tavassoli FA 1993 Proliferative and histologically malignant struma ovarii: a clinicopathologic study of 54 cases. Int J Gynecol Pathol 12:333[ndash]343. (4) Kostoglou-Athanassiou I, Lekka-Katsouli I, Gogou L, Papagrigoriou L, Chatonides I, Kaldrymides P. Malignant struma ovarii: report of a case and review of the literature. Hormone Research. 2002;58(1):34[ndash]38. (5) Roth LM, Talerman A. The enigma of struma ovarii. Pathology. 2007;39(1):139[ndash]146. (6)Makani S, Kim W, Gaba AR. Struma Ovarii with a focus of papillary thyroid cancer: a case report and review of the literature. Gynecologic Oncology. 2004;94(3):835[ndash]839. (7) Garg K, Soslow RA, Rivera M, et al. Histologically bland [ldquo]extremely well differentiated[rdquo] thyroid carcinomas arising in struma ovarii can recur and metastasize. (8) Roth LM, Karseladze AI. Highly differentiated follicular carcinoma arising from tstrume ovarii: a report of three cases and review of the literature, and a reassessment of so-called peritoneal strumosis. Int J Gynecol Pathol 2008; 27(2):213-22.[br][br]Nothing to Disclose: BNB, JMP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1318 63 494 SAT-405 PO56-01 Saturday 433 2012


434 ENDO12L_SAT-406 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Coexistence of Malignant Struma Ovarii and Cervical Papillary Thyroid Carcinoma (PTC) Aaron Leong, Miltiadis Paliouras, Louise Rochon, Mark Trifiro, Michael Tamilia Jewish General Hospital/McGill University, Montr[eacute]al, Canada; Jewish General Hospital/McGill University, Montr[eacute]al, Canada; Jewish General Hospital/McGill University, Montr[eacute]al, Canada [bold]Background:[/bold] Struma ovarii is an uncommon monodermal teratoma where thyroid tissue is the predominant element. Malignant transformation of struma ovarii is an even rarer occurrence.[br][bold]Clinical case:[/bold] We describe a case of a previously healthy 42-year-old woman who underwent a total abdominal hysterectomy/bilateral oopherectomy for a symptomatic left pelvic mass. Pathology identified malignant struma ovarii with classical papillary thyroid carcinoma (PTC) expression. There was no evidence of invasion through the ovarian cortex however analysis of the peritoneal fluid retrieved thyroid-type tumor ovarian cells. In light of these findings, an ultrasound of the cervical neck was performed which merely showed thyroid micronodules. Nonetheless, as the ovarian tumor was large, the patient underwent a total thyroidectomy with the intention of administering adjuvant I-131 therapy. Pathology of the cervical thyroid gland revealed bilateral multifocal PTC with extrathyroidal extension and perithyroidal lymph node metastasis. In spite of this, radioiodine post-therapeutic scan and PET-CT scan did not disclose advanced disease linking these two tumor sites and the stimulated thyroglobulin level remained low suggesting an excellent prognosis.[br][bold]Clinical Interpretations/Lessons:[/bold] This coexistence of malignant struma ovarii and cervical PTC could either represent a primary thyroid carcinoma with metastasis to the ovary or synchronous PTC in two distinct sites. We discuss the various clinical features (cancer dissemination characteristics) and tumor phenotypic and genotypic profiles to support the independent existence of these two embryologically related, although topographically distinct malignancies. Therefore, we report on the morphological (microscopy), immmunohistochemical (Hector Battifora Mesothelial Cell 1 (HBME-1), Cytokeratin-19 (CK-19), Galectin-3) and molecular (BRAF V600E, RAS, RET-PTC) results. In addition, through Human androgen receptor gene (HUMARA) X-chromosome inactivation analyses of the normal and malignant tissues, we hope to gain insight on the clonal origin of multifocal PTC.[br][bold]Conclusion[/bold]: We explored the clinical variables and molecular determinants of the coexistence of malignant struma ovarii and cervical PTC to make novel recommendations on its work-up, management and prognostication. This unique case could potentially clarify our understanding on thyroid tumorigenesis. To our knowledge, this coexistence has not been described in the medical literature.[br][br]Nothing to Disclose: AL, MP, LR, MT, MT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2220 63 495 SAT-406 PO56-01 Saturday 434 2012


435 ENDO12L_SAT-407 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Occult Thyroid Cancer with Distant Metastasis Tulsi Sharma, Roberto Izquierdo State University of New York (SUNY) Upstate Medical University, Syracuse, NY Introduction: Papillary thyroid carcinoma (PTC) generally carries a good prognosis since it usually remains intrathyroidal and tends to metastasize locally to lymph nodes alone. Distant metastases occur to the lungs and bone, and these are very rare especially from an occult primary.[br]Case: A 21-year-old lady presented to an outside facility in October 2008 after a motor vehicle accident. CT thorax was negative for any traumatic injuries; however, it revealed a diffuse reticulonodular pulmonary process. Sputum for AFB and PPD were negative and she was asymptomatic. She did not return for her follow-up appointments and returned to her PCP in November 2009 for an annual evaluation. Repeat CT thorax revealed a stable nodular pattern.[br]She was referred to Upstate Medical University in early 2010 with possible diagnosis of sarcoidosis. She still denied any constitutional symptoms and clinical exam was normal. Repeat CT-thorax revealed persistence of the miliary pattern. Blood work, ACE-level and PPD were negative. What would cause these numerous nodules which have been stable for 2 yrs and without any clinical manifestations?[br]After a detailed discussion the patient agreed for a bronchoscopy. Bronchoscopy with transbronchial biopsy revealed metastatic well-differentiated PTC! Thyroid sonography revealed a small solid nodule in the left thyroid lobe with internal and peripheral vascularity. The patient underwent a near-total thyroidectomy and a limited central compartment neck dissection. She has since undergone radioactive iodine therapy and is on levothyroxine. Her follow up hypothyroid I131 whole body scan showed a significant decrease in uptake in the lungs.[br]Discussion: Even in the presence of metastatic disease survival periods for PTC may exceed 20 years. The lung is the most common site for distant spread. Diffuse lung metastatic disease from an occult thyroid cancer is however extremely rare.[br]The growth rate of pulmonary nodules is often used to help differentiate benign from malignant disease. Pulmonary nodules that exhibit lack of growth for more than two years are generally considered benign. However, nodular lung metastases from PTC are an exception and may demonstrate lack of significant growth over many years. The cause of this growth arrest is unknown.[br]Conclusion: 1.Distant metastatic disease is extremely rare from an occult PTC.[br]2.Pulmonary metastases from PTC may have prolonged periods of growth arrest.[br][br](1) Chariot P, et al. Chest. 1993 Sep;104(3):981-2. (2) Zimmerman D, et al. Surgery 1988; 104:1157-66. (3) Kressel HY, et al. West J Med. 1978;129(5):424-9.[br][br]Nothing to Disclose: TS, RI 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 726 63 496 SAT-407 PO56-01 Saturday 435 2012


436 ENDO12L_SAT-408 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Spindle Epithelial Tumor with Thymus-Like Differentiation (SETTLE) in a 12-Year-Old Male Paulo Cesar Alves Silva, Paulo Roberto Fortunato Nascimento, Juliana van de Sande Lee, Genoir Simoni, Edson Cechinel, Rose Marie Muller Linhares, Jalmir Rogerio Aust, Daniella Serafin Couto Vieira, Marilza Leal Nascimento Hospital Infantil Joana de Gusm[atilde]o, Florian[oacute]polis, Brazil; Universidade Federal de Santa Catarina, Florian[oacute]polis, Brazil; Universidade Federal de Santa Catarina, Florian[oacute]polis, Brazil; Universidade Federal de Santa Catarina, Florian[oacute]polis, Brazil Background: Spindle Epithelial Tumor with Thymus-like Differentiation is a rare malignant neoplasm occurring predominantly in children, adolescents and young adults. It was first described as a cervical tumor derived from ectopic thymic tissue or remnants of the branquial pouch. SETTLE seems to behave as a low-grade malignancy, with potential of sending late metastases. Therewith patients should receive special attention in the initial diagnosis and long term follow up after resection of the lesion.[br]Case report: We present a male adolescent who presented at 12 yo of age to our clinic due to cervical nodule and right cervical lymphadenopathy. Laboratory testing revealed normal thyroid function. Neck ultrasound showed a hypoechoic nodule, with irregular borders and gross calcifications within, measuring 2.2 x 1.4 x 1.8 cm in the right lobe of the thyroid and bilateral cervical lymph nodes. The patient subsequently underwent fine needle aspiration biopsy of the thyroid nodule, with pathologic criteria for medullary thyroid carcinoma, spindle cell variant. Total thyroidectomy and resection of cervical lymph nodes were performed, in which confirmed by histological findings and immunohistochemistry study the diagnosis of SETTLE.[br]Conclusion: A long term follow-up is recommended since these patients may present with delayed metastases. With early diagnosis and prompt surgical intervention, this patient would be expected to have a favorable prognosis.[br][br]Nothing to Disclose: PCAS, PRFN, JvdSL, GS, EC, RMML, JRA, DSCV, MLN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 392 63 497 SAT-408 PO56-01 Saturday 436 2012


437 ENDO12L_SAT-409 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Multiple Primary Malignancy: Synchronous Thyroid Papillary Cancer and Astrocytoma [mdash] Case Report Albert D Makarov, Lyubov N Korosteleva, Violetta I Arzhakova, Elena R Makarova National Center for Medicine, Yakutsk, Russian Federation The incidence of multiple primary cancers is reported to be between 0.3% and 4.3%. The second primary lesion is identified either simultaneously with the primary lesion (synchronous) or after a period of time (metachronous). We report such a rare occurrence of dual malignancy of the thyroid and brain in order to stress the importance of a good preoperative workup to arrive at such a diagnosis preoperatively and also to stress the importance of radical surgery in patients with operable primary tumors. A 47-year- old man was admitted to our Medical center on March 18, 2010 with 4-month history of weakness of the right fingers, with a thyroid mass. Magnetic resonance imaging revealed a mass lesion in the right temporal, frontal lobes of brain with diffuse high intensity on T2-weighted and fluid-attenuated inversion images. A few small lesions were enhanced by contrast medium on the T1-weighted images. The thyroid gland was evaluated by ultrasonography, which demonstrated nodules (size 2,6x2,1x2,0cm and 5 mm) in the right lobe and 3 nodules 4 mm, 6,mm, 8 mm) in the left lobe. Ultrasonography guided fine-needle aspiration biopsy was performed and the cytological examination revealed cells of papillary thyroid cancer with metastases to the lymph nodes. The absence of familial adenomatous polyposis was determined after a complete colonoscopy failed to reveal any other colorectal pathology. In 24/03/2010 subtotal removal of brain tumor was performed. Histological examination: the brain neoplasm was astrocytoma grade II according to the WHO. In 5/04/2010 a total thyroidectomy with central and unilateral modified left radical neck dissection on the was performed.Histology demonstrated a papillary thyroid carcinoma. Postoperatively, he underwent radioactive iodine ablation therapy. In order to explain the molecular basis for such an occurrence, we performed the immunohistochemistry of the operative specimens. The cells of papillary thyroid cancer reacted for p53 - (+) in 75% sells, (HER-2 neu)-(+2 in 90% cells), vimentin - (+), EMA-(+), L-thyroglobulin (+), estrogen receptors -6F11 (+),progesterone receptors-312 (++), pancytokeratin - (++). The astrocytoma was positive only for p53- (+) In the light of advances in molecular biology we can only conjecture that the mutant p53 which has been found in both papillary thyroid cancers and astrocytoma may be one such putative mutation underlying such an occurrence.[br][br]Nothing to Disclose: ADM, LNK, VIA, ERM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 71 63 498 SAT-409 PO56-01 Saturday 437 2012


438 ENDO12L_SAT-410 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) V600E BRAF Mutation Positive Anaplastic Thyroid Cancer in a Patient with Graves Disease Priscilla Pei Sze Chiam, Chiaw-Ling Chng, Lishya Liauw, Andrew Eik Hock Tan, Chan HL Norman Singapore General Hospital, Singapore, Singapore; Singapore General Hospital, Singapore, Singapore; Singapore General Hospital, Singapore, Singapore Background[br]Graves[apos] Disease (GD) is associated with a 7-10 times higher incidence of malignant nodule formation[sup]1[/sup] and possibly greater aggressiveness of the thyroid cancer. The reason for this is unknown. We present a rare case of GD associated with V600E BRAF mutation positive anaplastic thyroid carcinoma.[br]Clinical case[br]A 68 year-old lady with a history of GD in remission for the past 10 years was referred to our institution for a left neck mass and possible relapse of her GD. She noted a growing neck mass for the past 1 month, as well as hoarseness of voice for the past 3 months. There is no family history of thyroid cancers or previous head and neck irradiation. Examination revealed an enlarged, irregular hard left thyroid mass. Cervical lymph nodes were impalpable. She did not display any signs of thyrotoxicosis.[br]Laboratory tests revealed T4 level of 27.9 pmol/L (8.8-14.4), TSH of [lt]0.015 mU/L (0.65-3.70) and TSH receptor antibody levels of 16.1 IU/L (0.0-1.5).[br]Thyroid uptake scan showed a large hypofunctioning left solid-cystic thyroid mass measuring 3.1 x 3.2 x 3.1cm. Fine needle aspiration cytology (FNAC) revealed atypical follicular cells, suspicious for a follicular or papillary neoplasm. In view of the indeterminate FNAC result, a trucut biopsy of the left thyroid mass was done. Histology showed a poorly differentiated carcinoma.[br]CT scan of the neck showed the extension of the left thyroid mass into the strap muscles, oesophagus and trachea. The diagnosis of anaplastic thyroid cancer was made and the decision was for surgery in view of the intra-thyroidal disease.[br]She underwent a total thyroidectomy, bilateral tracheo-oesophageal groove clearance, sternal split and segmental tracheal resection. The final histology confirmed the diagnosis of anaplastic cancer with components of well differentiated papillary cancer. Further evaluation via genetic testing of this post-operative specimen revealed V600E BRAF mutation (c1799T[gt]A).[br]Post-surgery, she underwent radiotherapy and chemotherapy and is currently doing well 9 months after her initial presentation.[br]Conclusion[br]Our case highlights the importance of a low threshold for prompt and thorough investigation in a patient with a background of GD presenting with a thyroid nodule. Further research into the role of V600E BRAF mutation in contributing to the higher aggressiveness of thyroid cancer in GD is warranted.[br][br]1.Belfiore A, Russo D, Vigneri R, Filetti S (2001) Graves[apos]disease, thyroid nodules and thyroid cancer. Clinical Endocrinology 55: 711[ndash]718.[br][br]Nothing to Disclose: PPSC, C-LC, LL, AEHT, CHLN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 604 63 499 SAT-410 PO56-01 Saturday 438 2012


439 ENDO12L_SAT-411 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) A Large Anaplastic Thyroid Cancer with Metastasis: Long-Term Favorable Response to High-Dose External Beam Radiation Pinkson Sheila, Maureen Koops, Tripathy Devjit AL Murphy VA Hospital, San Antonio, TX; University of Texas Health Science Center, San Antonio, TX Background: Anaplastic thyroid cancer (ATC) is an extremely aggressive solid tumor that accounts for approximately 1.6% of all thyroid cancers in the United States. However, ATC accounts for more than 40-50% of deaths from thyroid cancer. No effective treatment exists and the median survival is only 5 months. We report a case of ATC in a 55 year old man who has survived ATC for 5 years.[br]Case: A 55 yr old Caucasian male presented with a rapidly enlarging neck mass. CT showed a large neck mass with tracheal compression and deviation. The left neck mass was 7 x 6.8 cm by thyroid ultrasound. Thyroid fine needle aspiration (FNA) showed large anaplastic cells with bizarre, pleomorphic nuclei and prominent nucleoli with a background of necrosis and immunohistochemical stains negative for Thyroid Transcription Factor-1 (TTF-1), thyroglobulin and cytokeratin AE1/3. The large mass was found unresectable and caused airway compromise; therefore, a tracheostomy tube was placed. Combination of chemotherapy and radiation was recommended. The patient refused chemotherapy but received fractionated external beam radiation therapy (XRT) (5000cGy). After receiving 2500cGy, his breathing improved and the patient removed his tracheostomy tube. Serial neck CTs demonstrated progressive decrease in tumor size. One year post XRT, neck mass was 2.5cm at greatest dimension by neck CT; however, a suspicious lung nodule was present. The biopsy demonstrated anaplastic cells. He underwent XRT to the lung mass. A PET/CT surveillance scan found recurrence in the neck and he received additional XRT (4440cGy). Patient remains alive and functional five years since diagnosis, and recent PET/CT found no evidence of recurrence or progression of the disease.[br]Conclusion: Most patients with ATC do not survive beyond one year of diagnosis. This is an unusual case of a large ATC with lung metastasis who had a favorable long-term response to high dose XRT with 5 year post diagnosis survival. Since chemotherapy has not improved survival, more data are needed to evaluate the role of high dose XRT alone as a treatment option for anaplastic thyroid cancer.[br][br]Nothing to Disclose: PS, MK, TD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2248 63 500 SAT-411 PO56-01 Saturday 439 2012


440 ENDO12L_SAT-412 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Metastatic Papillary Thyroid Carcinoma to the Kidney [mdash] 30 Years after Initial Diagnosis Ramona Dadu, Mouhammed Habra, Mimi I Hu Baylor College of Medicine, Houston, TX; University of Texas MD Anderson Cancer Center, Houston, TX [bold]Background: [/bold]Distant metastases (DMs) are seen in about 10% of patients with papillary thyroid carcinoma (PTC) during their disease course with the lungs and bones being the most commonly involved sites. Metastatic areas are usually found in the first decade after initial cancer diagnosis in the majority of patients. PTC with renal metastases is very rare with only few well described cases published to date. We present a case of PTC with DM to the kidney that was diagnosed more than 3 decades after initial diagnosis.[br][bold]Case presentation: [/bold]An 81 year-old woman was diagnosed with PTC follicular variant (unknown stage) at the age of 50 years and underwent total thyroidectomy and radioactive iodine followed by Levothyroxine therapy. Routine surveillance with thyroglobulin(TG) and ultrasound of the neck were not performed. Thirty years later, an 8.5 x 7.9 cm right upper pole renal mass was incidentally discovered on an abdominal ultrasound performed for an unrelated reason. The renal mass had increased FDG avidity (average 6.7 SUV) on PET/CT scan without other sites of metastatic disease identified. She underwent right nephrectomy for presumed renal cell carcinoma. Surgical pathology revealed an 11.5 cm well-circumscribed mass composed of variably-sized follicles with nuclear crowding and focal nuclear grooves. Immunohistochemical studies showed that the tumor cells were positive for TG, PAX 8 and TTF-1. These results supported the diagnosis of metastatic PTC follicular variant. Postoperative laboratory evaluation revealed TG level of 5.7 ng/mL, undetectable TG antibodies, TSH 0.34 uIU/mL (range: 0.27-4.2 uIU/mL), and free T4 1.74 ng/dL (range: 0.93-1.70 ng/dL). Ultrasound of the neck revealed two nonspecific subcentimeter nodules in the central compartment. The management included continued TSH suppression and follow up of TG, thyroid function tests and imaging studies of the neck and abdomen every 6-12 months.[br][bold]Conclusion: [/bold]This case illustrates the need for regular and long term follow up for thyroid cancer patients. Many cases with DMs can go undetectable initially in the absence of formal monitoring and surface later with gross DMs. Additionally, it highlights the indolent nature of many cases of recurrent thyroid cancer. Further investigation into prognostic determinants of recurrence risk for thyroid cancer is warranted to determine appropriate surveillance recommendations after treatment.[br][br]Nothing to Disclose: RD, MH, MIH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1141 63 501 SAT-412 PO56-01 Saturday 440 2012


441 ENDO12L_SAT-413 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Undifferentiated Follicular Thyroid Carcinoma with Rhabdoid Features: A Rare Pathologic Diagnosis Brittany Noel Bohinc, John C Parker Duke University Hospital, Durham, NC; Wilmington Health, Wilmington, NC CASE: This is a 74-year-old male who was found to have a right 2.4 cm thyroid nodule by ultrasonography. Fine needle aspiration (FNA) of the right thyroid lesion revealed cytologic atypia highly suspicious for medullary or anaplastic thyroid carcinoma, prompting total thyroidectomy. Pathologic diagnosis demonstrated a poorly differentiated thyroid malignancy of the right lobe. Nuclear heterogeneity by routine staining suggested features of medullary and/or anaplastic thyroid carcinoma to the initial pathologist. Immunohistochemical staining for calcitonin, chromogranin, estrogen receptor, and Congo Red (amyloid) were negative. The tumor was sporadically positive for synaptophysin, vimentin, and CK19. Further consultation was obtained with a final histopathologic diagnosis of poorly differentiated follicular carcinoma with rhabdoid features. Maximum tumor diameter was 1.6 cm with lymphovascular invasion but without extrathyroidal tumor extension. The patient was treated with 162 mCi of I-131 following Thyrogen-stimulation. Post-treatment whole body scan (WBS) revealed residual uptake in the thyroid bed but no evidence for distant metastasis. Thyrogen-stimulated thyroglobulin has remained undetectable over years of follow-up. The patient is doing well and is in remission greater than 5 years after initial diagnosis.[br]DISCUSSION: Undifferentiated follicuar thyroid carcinoma with rhabdoid features is a rare diagnosis. Many of the reported cases describe an aggressive phenotype appearing in middle-aged women resulting in death months to several years after diagnosis (1). The tumor is usually largely composed of undifferentiated follicular cells and 10-30% rhabdoid cells. Histopathologically, rhabdoid cells are typically large, with abundant cytoplasm, eosinophilic inclusions, and eccentric nuclei containing distinct nucleoli (2). Immunohistochemical staining is usually positive for vimentin, sarcomeric actin, myoglobin, and cytokeratin expression in the rhabdoid tumor cells. Additionally, the rhabdoid cells are usually negative for desmin, thyroglobulin, and calcitonin (3,4). Treatment is total thyroidectomy +/- lymph node dissection and radioactive iodine ablation. To our knowledge, this is the 11th known case of follicular carcinoma with rhabdoid features and the longest known patient survival after treatment.[br][br](1) Agarwal S, Sharma MC, Aron M, et al. Poorly differentiated thyroid carcinoma with rhabdoid phenotype: a diagnostic dilemma[mdash]report of a rare case. Endocr Pathol. 2006; 17(4): 399-405. (2) Lai ML, Faa G, Serra S, et al. Rhabdoid tumor of the thyroid gland: a variant of anaplastic carcinoma. Arch Pathol Lab Med 2005; 129(3): e55-7. (3) Albores-Saavedra J, Sharma S. Poorly differentiated follicular thyroid carcinoma with rhabdoid phenotype: a clinicopathologic, immunohistochemical and electron microscopic study of two cases. Mod Pathol 2001; 14(2): 98-104. (4) Chetty R, Govender D. Follicular thyroid carcinoma with rhabdoid phenotype. Virchows Arch 1999; 435(2): 133-6.[br][br]Nothing to Disclose: BNB, JCP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1620 63 502 SAT-413 PO56-01 Saturday 441 2012


442 ENDO12L_SAT-414 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Anaplastic Thyroid Cancer: The Potential Role of TSH Stimulation Monika Datt, Rajshree Patel, Alice Lee, Salini C Kumar, Kenneth H Hupart NuHealth, Nassau University Medical Center, East Meadow, NY; St John[apos]s University, Queens, NY; Albert Einstein College of Medicine, Bronx, NY ATC remains one of the most lethal of human malignancies. We present a patient who developed anaplastic cancer after a decade of overt hypothyroidism.[br]A 59 yo female with papillary DTC underwent a total thyroidectomy in 2001. A 6 cm tumor with extracapsular and vascular invasion was present as were metastasis to bilateral LN. She received 154mCi of I-131 and was classified as T3N1bM0, stage 4a. Whole body scans from 2003-2009 revealed no metastases, however thyroglobulin levels were positive with fluctuating concentrations as high as 63. Pt had erratic compliance with appointments and l-T4 treatment and never achieved TSH suppression (TSH 30-130 uIU/mL). In 2009, a right sided neck mass was noted with a benign fine needle aspiration. 6 months later the mass grew and became painful. FNA was positive for cancer. Pt underwent excision in June 2011 revealing ATC. PAX8 and p53 mutation stains were positive. She underwent chemotherapy and radiation treatment for about 1 mos before choosing palliative care.[br]21-79% of ATC cases had coexisting or prior history of DTC. T4 therapy to achieve a low or suppressed TSH is commonly employed to decrease the likelihood of recurrence or progression. For more than a decade our patient[apos]s serum TSH was commonly in the overt hypothyroid range. This suggests the possibility that our patient[apos]s prolonged TSH elevation was important in the progression and dedifferentiation of her cancer. This concept has been explored in situations involving both TSH elevation and in patients with Graves[apos] disease who have displayed persistent titers of thyroid stimulating immunoglobulin (TSIg). In vitro, human ATC cell lines have demonstrated the presence of CD-133, a transmembrane protein which is expressed in stem cells. Friedman et al1 demonstrated that these cells proliferate with TSH receptor stimulation, and TSH induces increased expression of TSH receptor protein. This provides a framework for understanding how TSH suppressive therapy may play a role in prolonging survival in patients with differentiated thyroid cancer.[br]At the time of her original diagnosis, our patient had significant risk for a poor outcome. Nonetheless, her decade of overt hypothyroidism raises the possibility that TSH played a role in her tumor progression. As we explore DTC treatment strategies defined by individual patient risk stratification, our patient may suggest an area of inquiry that can help decide whether TSH suppression should be pursued.[br][br]Friedman, S et al. PLoS One,4:e5395, 2009.[br][br]Nothing to Disclose: MD, RP, AL, SCK, KHH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2355 63 503 SAT-414 PO56-01 Saturday 442 2012


443 ENDO12L_SAT-415 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Papillary Thyroid-Type Carcinoma Arising from Struma Ovarii. A Case Report Jerome Rebollos Barrera, Frances Lina Lantion Ang University of the Philippines-Philippine General Hospital, Manila City, Philippines Struma Ovarii and its malignant transformation to thyroid-type carcinoma are rare conditions. In recent reviews of reported literatures, only 53 cases of papillary thyroid-type carcinoma arising from struma ovarii were documented from 1924 to 2008 (1). Due to the rarity of the disease, lack of uniform histological criteria for malignancy and protracted clinical course, its management is not also universally accepted by physicians. The aim of this paper is to present a very rare case of papillary thyroid-type carcinoma arising from struma ovarii, and review literatures on the suggested consensus for diagnosis and management of the case.[br]A 41-year old Filipino woman presented to our section after she underwent total abdominal hysterectomy with bilateral salphingectomy for suspected primary ovarian carcinoma. She has been complaining of vague abdominal pain and distention for almost a year. Her pre-operative CT scan of the abdomen revealed large cystic mass with calcification suggestive of ovarian teratoma. Intra-operatively, the left ovary was converted to an 11 x 8 x 3.5 cm multiloculated, multiseptated mass densely adherent to the bowels and uterus. On pathologic examination with thyroglobulin staining, the large ovarian mass proved to be a papillary thyroid-type carcinoma arising from a struma ovarii. She underwent platinum-based chemotherapy used for ovarian teratoma.[br]On review of available case reports and case series done on papillary thyroid-type carcinoma, surgical resection of the tumor has been shown as effective treatment modality. Total thyroidectomy, to facilitate whole body scan and rule out primary thyroid carcinoma, radioactive iodine ablation, and thyroxine suppression therapy have shown to significantly decrease tumor recurrence and mortality (2). Chemotherapy and radiotherapy did not show any benefits on clinical outcomes.[br]Our patient underwent total thyroidectomy, radioactive ablation therapy and levothyroxine suppression therapy after tumor resection.[br]Papillary thyroid-type carcinoma arising from stuma ovarii as presented in our case should, therefore, be diagnosed and managed as primary papillary carcinoma of thyroid gland.[br][br](1)Roth, LM et al., Papillary Thyroid-type Carcinoma Arising in Struma Ovarii. A report of 4 Cases and Review of Literature. (2)Mircescu, H et al.,Papillary Carcinoma within Struma Ovarii: A Case Report with Literature Review.[br][br]Nothing to Disclose: JRB, FLLA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1103 63 504 SAT-415 PO56-01 Saturday 443 2012


444 ENDO12L_SAT-416 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) An Unusual Finding of Two Distinct Malignancies Concomitantly Found in Thyroid: Metastatic Malignant Melanoma and Papillary Thyroid Carcinoma Ongkarn Sarasombath, Uzma Khan University of Missouri, Columbia, MO Thyroid gland is an uncommon site of malignant metastasis, especially from malignant melanoma. We report a rare case of concomitant finding of metastatic malignant melanoma and papillary thyroid carcinoma of the thyroid.[br]CASE: a 53 year-old female presented with rapidly enlarging right parotid mass. Past history was significant for a skin lesion resected from right anterior forehead 2 years ago with appearance concerning for melanoma, however pathology results were inconclusive. Biopsy of right parotid mass showed poorly differentiated metastatic carcinoma. An MRI also showed an incidental right thyroid mass. A thyroid ultrasound showed a hypoechoic right thyroid mass measuring 23.6 x 20.8 x 33.8 mm. An FNA showed papillary thyroid cancer. PET/CT showed evidence of activity in the parotid gland, lytic right iliac bone lesion, large active right thyroid lobe and multiple active bilateral supraclavicular, axillary, hilar, mediastinal and internal iliac lymph nodes. The patient underwent right parotidectomy, neck dissection, and right thyroid lobectomy. Pathology result reported evidence of melanoma metastatic lesion in the parotid gland (22 mm). Right thyroid lobe revealed metastatic melanoma (1.5 mm) and papillary thyroid cancer (6 mm). Two out of 16 lymph nodes were positive for papillary thyroid cancer. Completion thyroidectomy with lymph node dissection showed that her left thyroid had no malignancy. Two out of 9 lymph nodes had metastatic thyroid cancer. BRAF mutation was positive in melanoma tissue but negative in thyroid papillary carcinoma tissue. After thyroidectomy, the patient was placed on levothyroxine to maintain TSH around 0.1 mcunit/mL. After discussion with the patient, a decision was made to postpone radioactive iodine ablation until she has completed her management for melanoma.[br]DISCUSSION: A thyroid nodule in patient with history of malignancy should raise a question whether the mass is benign, a new primary, or a metastasis. Fine-needle aspiration biopsy can help differentiate them. This case showed that even though papillary thyroid cancer is common and melanoma in thyroid gland is rare, the finding of the mass in thyroid gland can be either one of them or can be both. It is very important to find out because the treatment is different. Clinically significant metastatic disease in the thyroid gland is a sign of poor long-term survival in a patient with melanoma because it is an indication of disseminated disease.[br][br]Nothing to Disclose: OS, UK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 954 63 505 SAT-416 PO56-01 Saturday 444 2012


445 ENDO12L_SAT-417 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Thyroid Cancer in Plummer Disease: A Report of Two Cases in the Philippines Queenie Guinto Ngalob, Iris Thiele Isip-Tan University of the Philippines-Philippine General Hospital, Manila, Philippines Background: Thyroid cancer with concomitant thyrotoxicosis is variably reported to be rare. Most foci of malignancy are small and occur as postoperative incidental finding in surgery for Graves[apos] disease, toxic multinodular goiter or toxic adenoma. Thyroid masses with clinical features of malignancy and concomitant thyrotoxicosis are less commonly reported.[br]Case 1: A 49 year old Filipina presented with a twelve year history of enlarging goiter. She also had a mass in the sternal notch which developed over one year. Examination revealed a large multinodular goiter measuring 12x12 cm and a 7x7 cm mass in the sternal notch. Investigations revealed follicular neoplasm on fine needle aspiration biopsy and T3 toxicosis. A 1.6 x 1.8 cm functioning nodule in the superolateral right lobe was demonstrated on thyroid scintigraphy. She underwent total thyroidectomy and excision of the manubrial mass. Histopathology showed follicular variant of papillary cancer with metastasis to the manubrium.[br]Case 2: A 71 year old Filipina presented with recurrent thyroid nodules of 21 year duration. She had three thyroid surgeries in the past. Histopathology of the last surgery was follicular cancer. She was lost to follow-up after each surgery and received no subsequent treatment. She again consulted for recurrence of the thyroid masses. She had no symptoms of obstruction, voice changes, hypothyroidism nor hyperthyroidism. Examination revealed a large multinodular goiter measuring 10 x 6 cm. Fine needle biopsy yielded follicular neoplasm. TSH was suppressed at 0.008 mIU/L (NV: 0.3-3.8). Free T4 was elevated at 36.2 pmol/L (NV: 11-24). Thyroid scintigraphy showed functioning thyroid tissues with a conglomerate size of 8 x 5.6 cm. Chest imaging revealed lung metastasis. She is presently being treated with Methimazole 40 mg daily in preparation for completion thyroidectomy.[br]Conclusion: Plummer[apos]s disease may harbour cancer and behave aggressively.[br][br]Nothing to Disclose: QGN, ITI-T 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1421 63 506 SAT-417 PO56-01 Saturday 445 2012


446 ENDO12L_SAT-418 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Bilateral Vocal Cord Paralyses Secondary to Metastatic Papillary Thyroid Cancer Catherine HY Yu, Jennifer Anderson University of Toronto, Toronto, Canada; St Michael[apos]s Hospital, Toronto, Canada; St Michael[apos]s Hospital, Toronto, Canada Objective: To describe a case of papillary thyroid cancer (PTC) and bilateral vocal cord paralyses secondary to carotid sheath and vagus nerve (VN) invasion, and discuss intraoperative consideration of bilateral VN paralyses and its implications on voice, airway, swallowing and cardiovascular physiology.[br]Case: A 66-year-old woman presented with an 8-month history of progressive voice change, dyspnea, dysphagia and weight loss. Exam revealed a thin woman with inspiratory stridor, a weak breathy voice and a hard nodule on the posterior aspect of the left thyroid lobe. Indirect laryngoscopy showed bilateral abnormal vocal fold movement (complete paralysis of left, slight adduction of right) with pooled secretions in pyriform fossae. Computed tomography (CT) demonstrated a 1.3 cm nodule (left lobe), 2.7 cm mass (below right inferior pole) and bilateral lymphadenopathy in zones 3, 4 and mediastinum. Fine needle aspiration biopsy of a left sided lymph node showed PTC. The patient underwent total thyroidectomy, bilateral functional neck dissection of zones 2 to 6, and a planned tracheotomy for airway management. Intra-operatively, left-sided lymphadenopathy was grossly invading the carotid sheath. Given extensive VN involvement, it was sacrificed. On the right, lymphadenopathy was similar with carotid sheath invasion; however the tumor was less adherent and shaved off the VN in order to avoid the morbidity of bilateral VN paralyses. Pathology confirmed mixed classical and follicular variant of PTC replacing the left lobe with perineural and extrathyroidal invasion. Ten of 24 nodes had PTC with extranodal and vascular invasion. She received radioactive iodine ablation (200 mCi); body scan showed intense activity in the thyroid bed only. Six months later, thyroglobulin fell from 1366 (stimulated) to 7.4 pmol/L (unstimulated) and CT showed reduction in mediastinal lymphadenopathy. Nine months post-operatively, she was successfully decannulated but continued to have a weak breathy voice and dysphagia.[br]Conclusion: Vocal cord paralysis is a rare manifestation of PTC more often seen with anaplastic cancer. Our patient[apos]s unique findings of dysphonia, stridor and dysphagia were due to bilateral lymphadenopathy with reduced abduction of cords due to VN involvement on the left and RLN involvement on the right (partially compensated by reduced adduction of cricothyroid muscles due to VN involvement) and reduced pharyngeal and cricopharyngeal constriction due to VN involvement.[br][br]Nothing to Disclose: CHYY, JA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2283 63 507 SAT-418 PO56-01 Saturday 446 2012


447 ENDO12L_SAT-419 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Medullary Thyroid Cancer Presenting as Bilateral Foot Drop Hasmik Arzumanyan, Andrea Tom Stanford University Medical Center, Stanford, CA; Santa Clara Valley Medical Center, San Jose, CA Background: Medullary Thyroid Cancer (MTC) accounts for [sim]4% of all thyroid cancer cases. 75-80% of MTC presents sporadically. It is autosomal dominant in [sim]20-25%.[br]Clinical Case: 42 y/o previously healthy Indian man presented to the ED with bilateral foot drop. He had no past medical problems. Family history was negative for cancer, HTN or hypercalcemia. Examination revealed limited dorsiflexion of 1/5 in bilateral lower extremities. Spine MRI revealed a T12 lytic lesion with pathologic fracture and compression. Urgent T10-L2 decompression and fusion was performed with red-tan tissue noted to wrap around the spinal cord. Surgical pathology was suggestive of MTC. Serum calcitonin level was 13838 pg/ml (nl 0-10), with elevated serum chromogranin A and CEA, and normal serum Ca, PTH, plasma metanephrines, TFTs. Staging CT revealed bilateral [lt]1cm thyroid nodules and multiple [lt]1cm pulmonary nodules. PET-CT revealed hypermetabolic T12 lesion, several rib lesions, left 8mm and right 1cm hypermetabolic thyroid nodules. Thyroid ultrasound (US) revealed right interpolar 1.6cm, left superior 1cm, and left interpolar 1.3cm nodules. US-guided FNA of left interpolar thyroid nodule was consistent with MTC, right thyroid nodule was suspicious for MTC. Genetic testing was positive for RET mutation (C620R). His 18 y/o son carried the same mutation. Son had calcitonin level of 39 pg/ml, [lt]1cm left thyroid nodule on US, and underwent total thyroidectomy with central compartment dissection showing C-cell hyperplasia and multifocal microcarcinoma.[br]Discussion: Inherited MTC syndromes affect approximately 1 in 30,000 individuals and consist of MEN 2A (Sipple[apos]s syndrome), FMTC, and MEN 2B. They are caused by germline autosomal-dominant gain-of-function mutations in the RET proto-oncogene. Our patient[apos]s mutation (C620R) is associated with MEN 2A and Familial MTC (FMTC). It confers ATA risk level B (levels A-D, level D-highest risk), MTC risk level 2 (VII International Workshop on MEN: levels 1-3, level 3-highest risk). As shown in our case, initial presentation of MTC can be of metastatic disease.[br]Conclusion: All people with MTC should be offered RET testing since [sim]1-7% of apparently sporadic MTC cases have RET mutations. 95% of MEN 2A and MEN 2B patients, and 88% of those with FMTC will have RET mutations. If index case is negative for RET mutation, there is [lt]1% chance of having hereditary MTC. Knowledge of the RET mutation can guide the timing of prophylactic thyroidectomy.[br][br]Kloos et al. Medullary Thyroid Cancer: Management Guidelines of the American Thyroid Association. THYROID. Volume 19, Number 6, 2009.[br][br]Nothing to Disclose: HA, AT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 791 63 508 SAT-419 PO56-01 Saturday 447 2012


448 ENDO12L_SAT-420 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Anaplastic Thyroid Carcinoma Presenting Clinically as Horner Syndrome Isabela Targas Inocencio Carvalho, Fadlo Fraige, Ricardo Ayello Guerra, Lenira Cristina Stella, Paula Paes Batista Silva, Gabriella Dias Arcari Benefici[ecirc]ncia Portuguesa Hospital, S[atilde]o Paulo, Brazil Introduction:The diagnosis of Horner[apos]s Syndrome is not necessarily linked to a malignancy, but it should be excluded. This syndrome is a rare complication of benign and malign thyroid diseases, and it can happens after thyroidectomy or preoperatively. This is usually caused by an intrathoracic goiter compressing the sympathetic plexus on its emergence from the chest.[br]Clinical Case: A 50 years-old woman, presented with weight loss of 8 kilograms in 4 months and hoarseness. Sudden painful increase of neck and right ptosis have appeared two months before.Without the complaints of diplopia, ocular burning or irritability. On physical examination, a hard, fixed increased right thyroid lobe (RL) was evident. Laboratory tests: Cr:0.9mg/dl(0.4-1.2);Cai:1.22 mMol/L(1.1-1.3); AP: 80U/L(40-130), serum calcitonin[lt]2pg/ml([lt]11.5);TSH:1.5mU/L(0.3- 4.5);FT4:1.63ng/dL(0.93-1.70);anti-TG:10U/ml ([lt]115) and anti-TPO:8.2U/mL([lt]34);LDH:696U/L (180-460);beta 2 microglobulin 2.49mg/L(0.60-2.45), protein electrophoresis normal.Neck computed tomography(CT): solid lesion at the base of the right side of the neck measuring 6,7x6,5x6,0 cm. The lesion displaces the trachea and esophagus to the left and surrounding the right common carotid artery in most of its extension. Part of the RL is in close contact with the tumor but it is not possible to define a cleavage plan. Neck CT 1 month after: increased lesion 6,2x6,8x8,0 cm. Thyroid U.S.: Expansive lesion with discrete Doppler vascularity with RI: 0.70.[br]Abdominal CT: liver with multiple hypodense oval formations. FNAB(Fine needle aspiration biopsy): presence of numerous atypical cells with scant cytoplasm and nucleus with granular chromatin singly arranged. Washed needle of FNAB: TG [lt] 0.2 ng/ml and calcitonin: 3.05 pg/ml. Tumoral incisional biopsy revealed undifferentiated malignant neoplasia, presence of epithelioid cells with scant cytoplasm and severe nuclear atypia. Identified markers in immunohistochemistry: AE1/AE3,vimentin and P53.The case is compatible with anaplastic thyroid carcinoma(ATC).[br]Conclusion: This rare case is a ATC with atypical clinical presentation of Horner[apos]s Syndrome.Its very aggressive behavior can be attributed, according to the literature, to the overexpression of p53, MIB-1 and topoisomerase 2 alfa.The washed needle of FNAB with negative TG rule out the possibility of the case be a differentiated tumor and also negative calcitonin discards medullary thyroid carcinoma.[br][br](1) Smallridge RC, Copland JA. Anaplastic thyroid carcinoma: pathogenesis and emerging therapies. Clin Oncol (R Coll Radiol). 2010 Aug;22(6):486-97. Epub 2010Apr 24. Review. PubMed PMID: 20418080. (2) Chiacchio S, Lorenzoni A, Boni G, Rubello D, Elisei R, Mariani G. Anaplastic. thyroid cancer: prevalence, diagnosis and treatment. Minerva Endocrinol. 2008. Dec;33(4):341-57. Review. PubMed PMID: 18923370. (3) Broome JT, Gauger PG, Miller BS, Doherty GM. Anaplastic thyroid cancer. manifesting as new-onset Horner syndrome. Endocr Pract. 2009 Sep-Oct;15(6):563-6. Review. PubMed PMID: 19491063.[br][br]Nothing to Disclose: ITIC, FF, RAG, LCS, PPBS, GDA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 895 63 509 SAT-420 PO56-01 Saturday 448 2012


449 ENDO12L_SAT-421 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) PET-CT in the Detection of Differentiated Thyroid Carcinoma Residual Disease Lavinia Magdalena Vija, Taly Meas, Isabelle Faugeron, Laetitia Vercellino, Marie-Elisabeth Toubert St Louis Hospital, Paris, France Background:[br]Positron emission tomography (PET)/computed tomography (CT) with fluorodeoxyglucose (FDG) is at present increasingly used in early detection of residual disease or recurrences of differentiated thyroid cancer (DTC) patients who present with increasing thyroglobulin (Tg) levels and a negative 131I whole-body scan (WBS).[br]We report two patients with persistent disease due to tracheal and laryngeal invasion, detected only by FDG-PET. Appropriate identification of the localisation of the thyroid cancer remnants allowed a proficient treatment and a long-term adequate management.[br]A 62 year old patient with recent dysphonia was discovered to present a large calcified tumor infiltrating the right cervical region up to the upper side of the right clavicle on a cervical and thoracic CT, which was diagnosed as papillary thyroid carcinoma on fine needle aspiration biopsy. Total thyroidectomy with right lymph node dissection confirmed a 3mm tall-cell follicular thyroid cancer within the right thyroid lobe invading adjacent adipose tissue, oesophageal muscles, adjacent nervous fibers and local lymph nodes (pT4N1bM0). 3700 MBq 131I post-operative WBS revealed only left cervical remnant foci (Tg=18 ng/ml). Six months later, stimulated Tg was still detectable at 10 ng/ml, justifying a second RI treatment: post therapeutic RI WBS was negative, whereas concomitant PET/CT revealed an intense FDG uptake on the right thyroidectomy bed, corresponding to a lesion deviating and compressing the trachea. External radiotherapy was performed on the cervical peri-operatory areas with very good results, as 3 years later FDG-PET scan was normal, with undetectable stimulated Tg levels.[br]A 53 year old woman treated by total thyroidectomy followed by RI therapy for multifocal papillary thyroid cancer (pT4N1bM0) presented rhTSH stimulated detectable Tg (17.3 ng/ml) with negative RI WBS 6 months after RI ablation. FDG-PET revealed a unique hypermetabolic erosive lesion of the left lateral part of the trachea. Partial left laryngectomy with left lateral lymph node dissection was thus performed, confirming papillary localisation. Three year follow[ndash]up confirmed complete resection of residual disease as both stimulated Tg levels and FDG-PET were negative.[br]Conclusion: FDG-PET-CT is a very useful method to detect residual disease in DTC patients.[br][br]Nothing to Disclose: LMV, TM, IF, LV, M-ET 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2141 63 510 SAT-421 PO56-01 Saturday 449 2012


450 ENDO12L_SAT-422 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) LORAN: An Occupational Risk for Thyroid Cancer? Peter McIntyre, Robert Vigersky Walter Reed National Military Medical Center, Bethesda, MD From 1942 to 2011 the US Coast Guard operated over 150 LOng RAnge Navigation (LORAN) stations in the US, Italy, Japan, and Turkey which were the official radio-navigation system for both military and public use. An estimated 10,000 Coast Guard personnel operated and maintained this equipment until the final station closed in March 2011. Periodic investigations showed typical radiation exposures below occupational limits, so LORAN operators were not routinely monitored for radiation exposure despite ionizing radiation generated by their equipment. In response to concerns from LORAN veterans, many of whom have developed head and neck cancers, the Coast Guard released a report in March 2011 confirming that certain individuals performing [ldquo]exceptional operational and maintenance procedures[rdquo] may be at increased risk for radiogenic disease such as cancer.[br]Case Report:[br]The patient is a 63 year old Caucasian man who served in the US Coast Guard as a LORAN operator from 1971-1974. In addition to usual LORAN operation, he had recurring high risk exposures which deviated from usual protocol. He and colleagues routinely disposed of glass vacuum tubes containing Cobalt[sup]60[/sup], Nickel[sup]63[/sup], Krypton[sup]85[/sup], and/or Radium[sup]226[/sup] by breaking them to scavenge for copper. He was diagnosed with Stage 1 papillary thyroid cancer in 1977. Since then he has developed regional lymph node and pulmonary metastases. Most recently, he suffered a supraspinatus nerve injury during excisional biopsy of a posterior cervical lymph node which ultimately was found to be benign. His neurologic impairment has limited his occupational and recreational function. Currently, he is without evidence of active disease.[br]Conclusion:[br]Our patient may represent an example of LORAN-induced thyroid cancer despite the short latency period. There is a paucity of data about latency in occupational exposures and virtually none about exposure to LORAN-induced radiation. The latency between radiation exposure and the development of thyroid cancer is complex - age, dose (including time, intensity and shielding), and frequency of exposure are risk factors. The short latency in our patient does not exclude a possible causal relationship since there was an increased incidence in thyroid cancer within the first 4 years after the 1986 Chernobyl accident. The experience of our patient emphasizes the importance of discussing occupational exposures particularly in those who served in one of the uniformed services.[br][br]Nothing to Disclose: PM, RV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 561 63 511 SAT-422 PO56-01 Saturday 450 2012


451 ENDO12L_SAT-423 POSTER SESSION: Thyroid Cancer Case Reports (1:30 PM-3:30 PM) Malignant Intra-Thyroidal Paraganglioma: Case Report and Review of Literature Sangeetha Radhakrishnan, Jamie Mitchell, Joseph Sharpf, Deborah Chute, Krupa B Doshi Cleveland Clinic, Cleveland, OH; Cleveland Clinic, Cleveland, OH; Cleveland Clinic, Cleveland, OH; Cleveland Clinic, Cleveland, OH; Louis Stokes Cleveland VA Medical Center, Cleveland, OH [bold]Introduction[/bold]: Intra-thyroidal paragangliomas (PG) are rare neuroendocrine tumors. Only 25 cases have been reported in literature[sup]1[/sup]. Most cases are confined within the thyroid capsule but few cases of locally invasive PGs have been reported. To our knowledge, no case of distant metastatic disease has been described. We report a case of locally invasive intra-thyroidal PG with possible liver metastasis.[br][bold]Case:[/bold] A 71 year old Caucasian female with h/o Hashimoto[apos]s hypothyroidism underwent bilateral neck exploration and sub-total parathyroidectomy for primary hyperparathyroidism and parathyroid gland hyperplasia. Prior to surgery, a fine needle aspiration (FNA) proven benign solitary right thyroid nodule measuring 2.53 x 2.64 x 4.02 cm was noted. Concurrent thyroidectomy was not performed. Within the next year, she complained of increased right neck fullness and difficulty in swallowing. Thyroid US showed that the nodule had increased in size to 3.56 x 4.03 x 7.91 cm and was causing left tracheal deviation. FNA showed possible follicular neoplasm. Right hemithyroidectomy and central neck dissection was performed. The mass was noted to encase the right recurrent laryngeal nerve and invaded the trachea and esophagus. Minimal residual disease on trachea and esophagus remained at the end of the surgery. Histologic and immunohistochemical exam showed an intrathyroidal PG with metastasis to one right central neck lymph node. Tumor cells were positive for synaptophysin and NSE and negative for chromogranin, calcitonin, CEA, CAM 5.2 and TTF-1. S-100 was negative within the tumor cells but highlighted sustentacular cells rimming of the tumor nests. Six months later, patient had biopsy proven recurrence when she developed new difficulties with swallowing and breathing. She underwent a left hemithyroidectomy, total laryngopharyngectomy and central lymph node dissection followed by radiation. Histologically, the PG invaded the trachea, fibroadipose tissue and lymph nodes. Six months later a new 4.2 X 2.9cm mass developed in the liver suspicious for metastatic disease, currently being worked up.[br][bold]Conclusion:[/bold] This case highlights the fact that intra-thyroidal PG must be considered in the differential diagnosis of solitary thyroid nodule. Literature review suggests that majority of these tumors are encapsulated or locally invasive at the time of diagnosis. To our knowledge, this is the first case of malignant intra-thyroidal PG with possible distant metastasis.[br][br](1)Ferri E et al.,Acta Otorhinolaryngol Ital 2009; 29:97.[br][br]Nothing to Disclose: SR, JM, JS, DC, KBD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1806 63 512 SAT-423 PO56-01 Saturday 451 2012


452 ENDO12L_SAT-424 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) The Impact of Thyroid Nodule Size on the Accuracy of Ultrasound-Guided Fine-Needle Aspiration Merica Shrestha, Barbara A Crothers, Henry B Burch Walter Reed National Military Medical Center, Bethesda, MD; Walter Reed National Military Medical Center, Bethesda, MD [bold]Background:[/bold] Thyroid fine needle aspiration (FNA) has reduced the number of patients referred to surgery for benign disease. False negative (FN) rates in the literature have ranged from 0.4-13%, but the effect of nodule size on the accuracy of thyroid FNA remains controversial. Due to concerns about sampling error, many patients with nodules [gt] 4 cm are referred for thyroidectomy. We retrospectively reviewed thyroid nodule FNA results and surgical outcomes in a large cohort of patients over a 10-year period.[br][bold]Methods:[/bold] All thyroid FNAs performed at the Walter Reed Army Medical Center September 2001-September 2011 were compared to pathology results in those referred for thyroidectomy. A strict correlation between the biopsy site, location and size of nodule on ultrasound and pathology report was ensured. FNA results were classified as benign, atypical, follicular neoplasm/suspicious for follicular neoplasm (SF), suspicious for malignancy (SM), or malignant according to the Bethesda System, and the pathology report was used to classify nodules as benign or malignant. Nodules were grouped by size: 0.5-0.9 cm (group 1), 1-3.9 cm (group 2), and [ge] 4 cm (group 3). Incidental thyroid cancer was not included.[br][bold]Results: [/bold]Of 3,013 patients undergoing FNA, 667 (22.1%) had surgery. Patients were excluded for nodules [lt] 0.5 cm, non-diagnostic FNA, or no preoperative ultrasound, leaving 540 patients with 695 nodules. FNA results were benign in 417 nodules (60%), atypical in 22 (3.2%), SF in 122 (17.6%), SM in 77 (11.1%), and malignant in 57 (8.2%). Postoperative malignancy rates by FNA result were 7% if benign, 4.5% if atypical, 23% if SF, 33.8% if SM and 78.9% if malignant. FNA accuracy was 60% in group 1, 68.5% in group 2, and 80.3% in group 3 (p = 0.01). The FN rates for FNA were 15.8% in group 1, 6.3% in group 2 and 7.1% in group 3 (p = 0.25). Sensitivity and negative predictive value were highest in group 2 at 81.6% and 93.7%, respectively. The prevalence of malignancy was 18.6% overall, 22.9% in group 1, 19.3% in group 2 and 14.2% in group 3 (p = 0.33).[br][bold]Discussion: [/bold]Our results show that nodule size [ge] 4 cm neither increases the risk of FN FNA results, nor the overall risk of malignancy. We show a tendency towards a higher FN rate in sub-centimeter nodules and conversely, a low FN rate in nodules [ge] 4 cm. Large nodule size should not prompt automatic referral for thyroidectomy and an increased awareness of potential sampling error in sub-centimeter nodules is warranted.[br][br]Nothing to Disclose: MS, BAC, HBB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 563 64 513 SAT-424 PO41-01 Saturday 452 2012


453 ENDO12L_SAT-425 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Long-Term Growth Rate of Thyroid Nodules Monitored by Ultrasound Laura Middleton, Donald L Bodenner, Ann T Riggs, Spencer Pierson, Brendan Stack Winthrop P Rockefeller Cancer Institute, Little Rock, AR; University of Arkansas for Medical Sciences, Little Rock, AR; University of Arkansas for Medical Sciences, Little Rock, AR [bold]Introduction: [/bold]The aim of the present study was to evaluate the growth of thyroid nodules and examine factors that may influence growth such as initial nodule size, patient age, sex, race, weight, and TSH levels.[br][bold]Methods: [/bold]Thyroid nodules were measured by ultrasound over time by a single individual. Only nodules with a minimum of four measurements were included. Nodules were measured in 3 orthogonal planes and regression analysis was performed on the serial measurements of each orthogonal plane. Information gathered on the subjects included age, sex, race, weight, and TSH levels. Nodules in a multinodular gland were individually measured. Growth was defined as the largest change in any dimension over time.[br][bold]Results: [/bold]255 nodules in 125 patients were evaluated over an average of 4.7 years. Nodules were separated into categories based on their size at the initial ultrasound; 0 - 1 cm (90 nodules), 1-2 cm (108) nodules), 2-3 cm (33 nodules), and [gt]3cm (24 nodules). Neither patient demographics nor mean follow-up of nodules were significantly different among groups. Overall, 111 nodules (44%) increased in size although growth was slow in all size groups. Larger nodules grew faster than smaller nodules with a mean increase in size per year of 0.3 cm (+/-.003 cm/yr) in nodules 0 to 1cm compared with 0.99 cm per year (+/-.025 cm/yr) in nodules greater than 3 cm. 144 nodules decreased in size (56%), again at a very low velocity, ranging from.21 cm per year (+/-.003 cm/yr) in nodules 0 to 1 cm in size to.98 cm per year (+/-.024 cm/yr) in nodules greater than 3 cm in size. Most nodules that decreased in size were cystic or complex cystic. 116 nodules grew or shrank by less than one mm, within measurement error of the ultrasound instrument. The percent of nodules that did not grow were similar between initial size groups.[br][bold]Conclusions: [/bold]Thyroid nodules followed at our institution by serial ultrasound changed size very slowly, without regard to patient demographics. Similar number of nodules increased and decreased in size. Large nodules changed size faster than small nodules, but in absolute terms, the change was slow. Our results suggest that after initial evaluation and 6 month return, if no change is detected, ultrasound intervals may be extended to at least one year, and eventually to two to three years in most patients.[br][br]Nothing to Disclose: LM, DLB, ATR, SP, BS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 822 64 514 SAT-425 PO41-01 Saturday 453 2012


454 ENDO12L_SAT-426 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) The Value of Repeated US-Guided Fine-Needle Aspirations (US-FNAB) in the Follow-Up of Thyroid Nodules: Preliminary Results from the MoCyThy (Modena[apos]s Cytology of the Thyroid) Database Vincenzo Rochira, Anna Ansaloni, Serena Belli, Silvia Vezzani, Chiara Diazzi, Lucia Zirilli, Manuela Simoni, Cesare Carani, Bruno Madeo Unit and Chair of Endocrinology [amp] Metabolism, University of Modena [amp] Reggio Emilia, Azienda USL of Modena-NOCSAE, Modena, Italy INTRODUCTION. There is no consensus about the usefulness of repeating the US-FNAB during the follow-up of nodules when a benign (Thy2) or indeterminate (Thy3) report is obtained at first US-FNAB. AIM OF THE STUDY. To investigate the clinical value of repeating US-FNAB after a previous adequate Thy2 or Thy3 US-FNAB. METHODS. We reviewed the US-FNABs performed from 2006 to 2009. All clinical data of the patients were collected and analyzed using the MoCyThy DATABASE, which is the part of the institutional database ENDOBASE (based on the MySQL open source technology) devoted to store data of all institutional US-FNABs. Among 7983 records, we searched out 288 patients (327 nodules) undergoing at least two consecutive adequate US-FNABs for a total of 686 US-FNABs. We compared the first US-FNAB (Thy2 or Thy3) at baseline with the results of the following US-FNABs (2nd or 3rd US-FNAB). RESULTS. Of the 327 baseline US-FNABs, 58% were Thy2 and 42% Thy3. Of the 189 Thy2 at baseline, 157 (83%) were confirmed as Thy2 at follow-up, while 32 (17%) did not confirm the first diagnosis: 29 (15%) of them were Thy3 and 3 (2%) Thy4. No modifications of volume or US-features were recorded in these Thy4 from baseline. Of the Thy3 at baseline, 55 (40%) were confirmed as Thy3 at the follow-up, 84 (60%) did not confirm the first diagnosis: of them 6 (4%) were Thy4/5 and 78 (56%) Thy2. CONCLUSIONS. The outcome of a subsequent US-FNAB is often discordant compared with the first cytological diagnosis. A first cytological diagnosis of Thy2 does not completely exclude a malignant (Thy4) or an indeterminate (Thy3) lesion. A second US-FNAB after 6-12 months could be useful, in clinical practice, to definitively confirm benign lesions or to reduce the rate of malignant tumor or of follicular lesions unrecognized by the first US-FNAB.[br][br]Nothing to Disclose: VR, AA, SB, SV, CD, LZ, MS, CC, BM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 486 64 515 SAT-426 PO41-01 Saturday 454 2012


455 ENDO12L_SAT-427 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Thyroid Nodule Screening for Malignancy: A Prospective Study on Ultrasonographic/Clinical Profile, Cytology and BRAF V600E Mutation Evaluation Martina Rossi, Mattia Buratto, Stefania Bruni, Carlo Filieri, Federico Tagliati, Giorgio Trasforini, Roberta Rossi, Maria Donatella Beccati, Ettore degli Uberti, Maria Chiara Zatelli University of Ferrara, Section of Endocrinology, Ferrara, Italy; S Anna Hospital, Ferrara, Italy Ultrasound (US)-guided fine-needle aspiration biopsy (FNAB) is the most reliable nonsurgical test for distinguishing benign from malignant thyroid nodules. However, there is no consensus on which nodules should undergo FNAB, yet. The aim of our study is therefore to assess the diagnostic sensitivity of FNAB in a series of unselected thyroid nodules and to investigate the additional diagnostic contribution of BRAF V600E analysis in the detection of differentiated thyroid cancer. To thjs aim, 1856 patients with thyroid nodules displaying at least one US feature suspicious of malignant lesion were submitted to FNAB, for a total of 2421 cytoaspirates submitted to cytological evaluation and to biomolecular analysis.[br]We found that cytology has high positive predictive value and specificity for the diagnosis of malignant lesions. BRAF V600E mutation was found in 115 samples, 80 of which were also cytologically diagnosed as papillary thyroid cancer. BRAF mutation analysis significantly enhanced the diagnostic value of cytology, increasing FNAB diagnostic sensitivity for malignant nodules by 28%. Diameter was not a good predictor of thyroid nodule malignancy. Each investigated US/clinical characteristic of suspected malignancy correlated with the presence of a thyroid cancer in thyroid nodules with diameter of 4 mm or greater.[br]These data indicate that all nodules with at least one US/clinical characteristic of suspected malignancy should undergo FNAB. The diagnostic sensitivity for thyroid cancer is significantly increased by BRAF V600E mutation analysis, indicating that the screening for BRAF mutation in FNAB samples has a relevant diagnostic potential and should always be performed.[br][br]Nothing to Disclose: MR, MB, SB, CF, FT, GT, RR, MDB, EdU, MCZ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 485 64 516 SAT-427 PO41-01 Saturday 455 2012


456 ENDO12L_SAT-428 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Is There a Real Diagnostic Impact of Elastosonography and Contrast-Enhanced Ultrasonography in the Management of Thyroid Nodules with Indeterminate Cytology? Massimo Giusti, Lara Vera, Giulia Melle, Francesco Minuto, Tiziana Celiento, Enzo Silvestri, Gianni Turtulici AIRCCS AOU San Martino-IST, Genova, Italy; AIRCCS AOU San Martino-IST, Genova, Italy; Evangelico Hospital, Genova, Italy [bold]Objective[/bold] FNAB is the best single test in the management of thyroid nodules, but its potential presurgical value is limited by the finding of nodules with indeterminate cytological diagnosis (Thy3 according to BTA). Elastosonography (USE) and contrast-enhanced US (CEUS) have recently been introduced into the screening of thyroid nodules. The aim of this prospective study was to compare the sensitivity, specificity, positive predictive value (PPV) and accuracy of USE (ELX 2/1 index) and CEUS [Peak index and time to peak (TTP) index] in thyroid nodules with indeterminate cytology [bold]Design[/bold] 25 Thy3 nodules from 8 m and 17 f were studied in parallel with US areas of normal extra-nodular tissue. In all patients (pts), cytological-histological correlation was assessed. A presurgical US score was assigned. USE was evaluated according to the Esaote program and CEUS was performed with Bracco SonoView contrast medium. [bold]Results[/bold] Thyroid hormones were normal in all pts. Two pts were TPOAb+ and a slight increase in calcitonin levels was noted in 3 pts. The maximum nodule diameter was 22.4[plusmn]2.2 ml ([plusmn]SEM) and the cumulative US score was 2.4[plusmn]0.2. Malignancy was found in 14% of our Thy3 nodules, while in 9% of pts a thyroid microcarcinoma was found in extra-nodular tissue. No correlation between cumulative US scores and histology was found. There was no significant difference in the ELX 2/1 index (benign (b) 1.34[plusmn]0.14; malignant (m) 1.53[plusmn]0.14) in Thy3 nodules. After CEUS, Peak index (b 0.89[plusmn]0.08; m 0.69[plusmn]0.17) and TTP index (b 0.89[plusmn]0.05, m 1.07[plusmn]0.06) were similar. No correlation was found between histology and ELX1/2 index, P index or TTP index. USE displayed high sensitivity (100%) but low specificity (37%), PPV (23%) and accuracy (47%). After CEUS, the P index also showed very low specificity (15%), PPV (21%) and accuracy (31%). On the other hand, the TTP index of CEUS showed slightly better specificity (54%) and accuracy (62%) but always low PPV (33%). [bold]Conclusions[/bold] US and FNAB play a central diagnostic role in thyroid nodules. The US score does not modify the [ldquo]gray area[rdquo] of Thy3 nodules, in which malignancy can by found in about one in four pts. USE and CEUS are innovative techniques that need to be standardized. Our data on Thy3 nodules do not suggest a very promising role of USE. We cannot exclude a possible role of CEUS in the management of Thy3 nodules. A large number of pts with cytological-histological correlation are needed in order to obtain more conclusive results.[br][br]Nothing to Disclose: MG, LV, GM, FM, TC, ES, GT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 464 64 517 SAT-428 PO41-01 Saturday 456 2012


457 ENDO12L_SAT-429 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Ultrasound (US) Features of Thyroid Nodules with Cytology Suspicious for Malignancy Vincenzo Rochira, Erica Taliani, Anna Ansaloni, Chiara Diazzi, Giulia Brigante, Cesare Carani, Bruno Madeo Unit [amp] Chair of Endocrinology [amp] Metabolism, University of Modena [amp] Reggio Emilia, Azienda USL of Modena-NOCSAE, Modena, Italy Introduction: Several studies in literature have shown that some features of thyroid nodules at US are associated with malignancy. However, previous studies were focused mainly on subjects affected by multinodular goiter (about 70%) rather than subjects with thyroid cancer (about 30%). Furthermore, the main limitation of previous studies was the lack of thyroidectomy in all subjects.[br]Aim of the study: To evaluate the diagnostic value of US features in a selected sample of patients with thyroid nodules cytologically suspected for malignancy (THY4 [ndash] THY5) by comparing US features of each nodule with the results of histological analysis after thyroidectomy.[br]Methods: In this prospective study, we enrolled 54 patients with cytological result suspicious of malignancy. All subjects underwent thyroid ultrasound before thyroidectomy. We evaluated the following US features: size, content, shape, margins, echogenicity, calcification, halo sign, vascular pattern, for all the nodules (those citologically suspected and those not suspected). All enrolled patients underwent total thyroidectomy, therefore all benign and malignant nodules previously assessed at US received histological verification.[br]Results: In all the 54 patients a diagnosis of differentiated thyroid cancer was confirmed. Each of the following features: microcalcifications, macrocalcifications, irregular margins and hypoechogenicity at US correlate with malignancy at histology by using chi-square (p[lt]0.001).These features have high specificity but low sensitivity (microcalcifications 93.9% - 40.4%, macrocalcifications 98% - 22.8%, hypoechogenicity 96% - 21%, respectively). Irregular margins is the feature with the best pair of sensitivity (65%) and specificity (65%).[br]Conclusions: These results confirm and reinforce previous studies that showed a correlation among microcalcification, irregular margins, hypoechogenicity and malignancy in a highly selected sample of patients undergoing thyroidectomy. Furthermore, in contrast with literature, we found a strong correlation also between macrocalcification and malignancy. US is a valid tool to select which nodules require FNA evaluation according to sonographic features closely related to malignancy.[br][br]Nothing to Disclose: VR, ET, AA, CD, GB, CC, BM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 487 64 518 SAT-429 PO41-01 Saturday 457 2012


458 ENDO12L_SAT-430 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Densely Calcified Thyroid Nodules Have High Risk for Thyroid Cancer Jung Min Kim, Hee Kyung Kim, Jee Hee Yun, Soo Jung Kim, Ho-Cheol Kang Chonnam National University Medical Shool, Gwangju, Korea; St Carollo Hospital, Suncheon, Korea [bold]Backgrounds:[/bold] It has been controversial that dense calcifications in thyroid nodules are the high risk feature for thyroid cancers. The aim of the study was to evaluate the prevalence of malignancy in patients with densely calcified nodules with analysis of clinical parameters related to the development of thyroid cancer.[br][bold]Methods:[/bold] We retrospectively reviewed thyroid sonographic findings, results of fine-needle aspiration cytology (FNA) and clinical characteristics in patients with thyroid nodules who visited Chonnam National University Hwasun Hospital for 5 year period (2,387cases). Dense calcifications include [ldquo]egg-shell[rdquo] or rim-like peripheral calcification and coarse dense calcification (163 cases). Nodules with microcalcification ([lt]2mm) were excluded in this analysis (154 cases).[br][bold]Results:[/bold] Among the 163 densely calcified nodules, 45(27.6%) were malignant. Its prevalence was higher than that of patients without calcification (p [lt]0.01). Patients with densely calcified nodules were older (p [lt]0.01), and showed the higher frequency of obtaining inadequate FNA samples (1.2 % vs. 0.2 %) than patients without calcification. In densely calcified nodule group, malignant nodules (55.6%) were more often hypoechoic than benign nodule (37.2%).[br][bold]Conclusions: [/bold]Nodules with dense calcification carry a higher risk for malignancy, and higher risk for obtaining inadequate or non-diagnostic sample than non-calcified nodules. Special consideration should be endeavored not to misdiagnose malignancy in thyroid nodules showing dense calcifications on ultrasonography.[br][br]Nothing to Disclose: JMK, HKK, JHY, SJK, H-CK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 501 64 519 SAT-430 PO41-01 Saturday 458 2012


459 ENDO12L_SAT-431 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Utilization of Ultrasound-Guided Core Needle Biopsy as a Primary Diagnostic Technique for Evaluation of Thyroid Nodules, Compared to Fine-Needle Aspiration Biopsy Akshay Bhanwarlal Jain, Behrouz Salehian, Ellie Maghami, Elizabeth Scarlett Perry, Cheryl Torricelli, Bihong T Chen City of Hope National Medical Center, Duarte, CA; City of Hope National Medical Center, Duarte, CA; City of Hope National Medical Center, Duarte, CA; Wayne State University, Detroit, MI Objective: To study the efficacy of Ultrasound-guided core-needle biopsy (UG-CNB) in comparison to UG fine needle aspiration biopsy (UG-FNAB) as a primary diagnostic tool for evaluation of thyroid nodules.[br]Method: A retrospective chart review was performed on all patients undergoing first-time ultrasound guided biopsy of thyroid nodules from January 2007 through November 2011 at our institution. Both, UG-CNB and UG-FNAB are performed by the same group of radiologists. Biopsy results were then compared to surgical pathology reports of those patients undergoing hemi-thyroidectomy (HT) or total-thyroidectomy (TT), to determine accuracy of diagnosing malignancy with either UG-CNB or UG-FNAB.[br]Results: Of the 489 ultrasound guided biopsies, 399 were UG-CNB and 91 were UG-FNAB. Non-diagnostic results were seen in 6 (2%) of UG-CNB and 17 (19%) of UG-FNABs (p[lt]0.001; chi-square). Eighty-seven patients who had UG-CNB and 17 patients who had UG-FNAB (total=104) underwent HT or TT. The diagnostic accuracy of UG-CNB for detecting malignancy was 93% while that of UG-FNAB was 59% (p= 0.004; two-sided Fisher[apos]s exact test). Sensitivity and specificity of UG-CNB for detecting malignancy were 79% and 96%; for UG-FNAB were 14% and 90% respectively. Both, negative predictive value and positive predictive value were higher in UG-CNB compared to UG-FNAB- 88% and 93% versus 60% and 50% respectively. Both groups had one patient each having a localized hematoma post-procedure. No major complications were seen in either group.[br]Discussion: UG-FNAB is currently the diagnostic technique of choice for evaluation of thyroid nodules(1). However, FNAB is associated with non-diagnostic sampling in 10-20% of cases(2). FNAB has a limitation on follicular structure as specimens contain only cell aggregations and small tissue fragments, not the whole follicles. These limitations are thought to be overcome by use of UG-CNB, which provides a larger tissue sample that retains its cellular architecture. This may also increase the diagnostic yield of immunohistochemical markers of thyroid malignancy, such as HBME-1 and cytokeratin-19. Core biopsy can be particularly advantageous in centers where on-site cytology evaluation during the procedure is unavailable to determine adequacy of tissue sampling.[br]Conclusion: In comparison to UG-FNAB, UG-CNB is a more accurate technique for detection of malignant thyroid lesions, with no increase in the rate of complications.[br][br](1)Cooper DS et al., Thyroid. 2009 Nov;19(11):1167-214. (2)Gharib H et al., Ann Intern Med 1993; 118:282-289.[br][br]Nothing to Disclose: ABJ, BS, EM, ESP, CT, BTC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 875 64 520 SAT-431 PO41-01 Saturday 459 2012


460 ENDO12L_SAT-432 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) The Utility of Serum Calcitonin Measurement in Thyroid Nodule Evaluation: Experience of a Single University Hospital in Sao Paulo, Brazil Suemi Marui, Debora Seguro Danilovic, Rosalinda Yasato Camargo, Valeria Samuel Lando, Meyer Knobel, Marcelo Cidade Batista Disciplina de Endocrinologia-Faculdade de Medicina-Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Disciplina de Endocrinologia-Faculdade de Medicina-Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Routine calcitonin (Ct) measurements in patients with thyroid nodules have been recommended as a screening method for C-cell hyperplasia and medullary thyroid cancer (CMT). This approach may help in early detection and improve overall survival. Basal Ct [gt]100 pg/mL establish the diagnosis of CMT, so that surgery may be planned with cervical exploration. However, basal Ct [lt]100 may be seen in patients with C-cell hyperplasia and micromedullary carcinoma as well. In addition, normal cut-off Ct values vary among different assays. Objective: Analyze basal serum Ct levels in patients with thyroid nodules evaluated in a single University Hospital. Material and Methods: From 01/2009 to 01/2011, serum Ct was measured in all patients with thyroid nodules detected at ultrasound (US) in the outpatient clinic of Hospital das Clinicas-Universidade de Sao Paulo, Brazil. Serum Ct levels were determined using an automated solid-phase, enzyme-labeled, two-site chemiluminescent immunoenzymatic assay (Immulite 2000; Siemens Medical Solutions Diagnostic, UK). Ct[lt]2.0 pg/mL were considered undetectable. Thyroid US was performed in B mode with a scanner 7.5-12 MHZ transducer. Patients with normal thyroid parenchyma and familial CMT or MEN were excluded. Results: 134 patients (115 females) were selected, with mean age of 57 [plusmn] 17 yrs, from 17 to 95 yrs. All had solitary or multiple nodules at US and 68% underwent US-guided fine needle aspiration according to clinical decision. Cytology was benign (goiter) in 38; undetermined in 39; suspicious for papillary carcinoma in 2; and diagnostic of papillary carcinoma in 8 patients. In 15 patients with undetermined cytology submitted to thyroidectomy, histological diagnosis was follicular adenoma (3), papillary carcinoma (5) and goiter (7). No CMT or C hyperplasia was detected. Median Ct levels were 2.0 (maximum 7.0). In 11 patients with Ct[gt]2.1, a second measurement was [lt]2.0. Antithyroperoxidase antibodies were assayed in 25 patients with Ct [gt]2.1, but only 2 were positive. None had impaired renal function. Therapy with proton pump inhibitors was not compiled from patient[acute]s records. Conclusion: In our cohort, Ct measurements did not help in the diagnostic evaluation of thyroid nodules, since all patients had serum Ct [lt]7.0. Further studies in larger groups including patients with CMT and C-cell hyperplasia are needed to determine the diagnostic value of measuring Ct in patients with thyroid nodules.[br][br]Nothing to Disclose: SM, DSD, RYC, VSL, MK, MCB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 863 64 521 SAT-432 PO41-01 Saturday 460 2012


461 ENDO12L_SAT-433 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Comparison of Clinical Outcomes in Differentiated Thyroid Carcinoma between Childhood and Young Adult Patients Won Jin Kim, Min Jung Bae, Yang Seon Yi, Ji Hyun Kang, Bo Gwang Choi, Yun Kyung Jeon, Snag Soo Kim, Bo Hyun Kim, In Ju Kim, Yong Ki Kim Pusan National Univeristy Hospital, Busan, Korea; Kim Yong Ki Internal Medicine Clinic, Busan, Korea [bold]Objective[/bold]: The aim of the present study was to evaluate differences between childhood and young adults in presentation, clinical course and outcome of well differentiated thyroid carcinoma (DTC).[br][bold]Methods[/bold]: We retrospectively reviewed the medical records of 61 childhood and young adults (50 females, 11 males, [lt]25 years) with DTC who were treated with radioactive radioiodine (RI) and followed up between June 2002 and May 2010. All patients had undergone total thyroidectomy with lymph node dissection if enlarged lymph nodes were present and had been referred for initial radioiodine ablation. Recurrence-free survival (RFS) was evaluated with the Kaplan-Meier method.[br][bold]Results[/bold]: At diagnosis, extrathyroidal extension of DTC was more prevalent and average tumor size was bigger in the childhood than in the young adults ([italic]P[/italic]=0.045 and [italic]P[/italic]=0.002, respectively). However, there was no significant difference between 2 groups with regard to the presence of lymph node or distant metastases ([italic]P[/italic]=0.885 and [italic]P[/italic]=1.000, respectively). During follow-up, the recurrence in the thyroid bed or cervical lymph nodes occurred in 6 patients (20.7%) in the childhood and 3 patients (9.4%) in the young adults ([italic]P[/italic]=0.323). The RFS rate was similar in the childhood and young adult patients (log-rank test, [chi][sup]2[/sup]=2.424, df=1, [italic]P[/italic]=0.120).[br][bold]Conclusion:[/bold] Our result shows that although the presentation of DTC at the time of diagnosis was more aggressive in the childhood, intensive management elicited a similar clinical outcome in the childhood and in the young adults.[br][br]Nothing to Disclose: WJK, MJB, YSY, JHK, BGC, YKJ, SSK, BHK, IJK, YKK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 83 64 522 SAT-433 PO41-01 Saturday 461 2012


462 ENDO12L_SAT-434 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Histopathological Features and Tumor Size Relationship in Papillary Thyroid Cancer Ana Maria Chacon, Patrica Andrada, Cecilia Hernandez, Jose Ignacio Echeveste, Maria Dolores Lozano, Juan Carlos Galofre Cl[iacute]nica Universidad de Navarra University of Navarra, Pamplona, Spain; Cl[iacute]nica Universidad de Navarra University of Navarra, Pamplona, Spain Two of the main prognostic factors in papillary thyroid cancer (PTC) are tumor size (TS) and aggressive histopathological features (HF). However, it is not well established whether these two factors are interrelated. The aim of this study was to analyze the presence of different HF in PTC with several TS.[br]Data were retrospectively extracted from 115 PTC consecutive clinical records. The sample was divided by TS. We established a first cutoff at 1.0 cm and compared Group A: [ge]1cm (n=64) and Group B: [lt]1 cm (n=51). In a second analysis we set up a second cutoff point at 0.5 cm and then compared Group C [ge]0.5 cm (n=81) and Group D [lt]0.5 cm (n=34). And finally, we subdivided Group B into Group B1: [ge]0.5 cm (n=17) and B2 [lt]0.5 cm (which was identical to Group D) for the third analysis.[br]We compared the prevalence of the following HF between groups: mitotic activity, vascular invasion, extra-thyroidal extension, psammoma bodies, necrosis, capsule or pseudocapsule presence, and capsule invasion. Data were analyzed by chi-square test.[br]When the HF percentage values were compared between Groups A and B we found statistically significant differences in vascular invasion (23.4% vs. 5.8%; p[lt]0.01); extra-thyroidal extension (21.9% vs. 7.8%; p[lt]0.05); psammoma bodies (60.0% vs. 27.4%; p[lt]0.01); capsule presence (56.2% vs. 25.5%; p[lt]0.01); and capsule invasion (34.37 vs. 9.8%; p[lt]0.01), respectively. Similar results were obtained when Groups C and D were compared: vascular invasion (22.2% vs. 0%; p[lt]0.01); psamomma bodies (58.0% vs. 17.6%; p[lt]0.01); capsule presence (54.3% vs. 14.7%; p[lt]0.01); and capsule invasion (32.0% vs. 2.9%; p[lt]0.01), respectively. Finally, statistical significance difference was also found when comparing Groups B1 and B2 in terms of mitotic activity (23.5 % vs. 2.9%; p[lt]0.01); necrosis (11.7% vs. 0%; p[lt]0.01); capsule presence (47.0% vs. 14.7%; p[lt]0.05); and capsule invasion (23.5% vs. 2.9%; p[lt]0.05), respectively.[br]Overall, our results suggest that the presence of those histopathological features normally related with aggressiveness is higher in larger PTC tumors. In other words, this study establishes a link between tumor size and progressive development of aggressive histopathological features and suggests at least partially, why small variations of size are normally associated with different prognosis.[br][br]Nothing to Disclose: AMC, PA, CH, JIE, MDL, JCG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1523 64 523 SAT-434 PO41-01 Saturday 462 2012


463 ENDO12L_SAT-435 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Prevalence, Characteristics and Outcome of Tall Cell Papillary Thyroid Carcinoma: A Retrospective Cohort Study Maria F Russo Picasso, Carolina Carrizo, Ana Jaen, Marcelo Figari, Eduardo Mazzaro, Ana Mollerach, Andrea Kozak, Marta Balzaretti Hospital Italiano de Buenos Aires, Caba, Argentina; Hospital Italiano de Buenos Aires, Caba, Argentina; Hospital Italiano de Buenos Aires, Caba, Argentina Introduction: The reported prevalence of tall cell variant of papillary thyroid carcinoma (TCC) is between 3 and 19%, underscoring the wide regional differences and also the lack of consensus in the diagnostic criteria used. TCC has been associated with a more aggressive clinical course than other variants of papillary thyroid carcinoma (CPT)[br]Aims: To assess the prevalence of TCC in our medical centre and the clinico-pathological characteristics at presentation. Compare both presentation, and clinical outcome at 12months postsurgery[br]Design: Observational study in a 10 y retrospective cohort of patients operated for papillary thyroid carcinoma.[br]Materials and Methods: We reviewed the pathological and clinical records of patients operated for CPT in the Hospital Italiano de Buenos Aires between January 1st 2000 and December 31st 2010. TCC was defined as at least 30% of tall cells. These patients were compared to the rest of CPT (C). We used a stringent definition to consider a patient free of disease at 12 months postoperatively: Negative TSH-stimulated Tg, negative neck ultrasound and negative radioactive iodine uptake (RAIU).[br]Statistical analysis: Numeric variables were expressed as medians and interquartile ranges, and tested for differences with Mann-Whitney Test. Categorical variables are expressed as pecentages, and tested with Chi 2 test. p [lt]0.05 was considered statistically significant.[br]Results:. From January 1st 2000 and December 31st 2010, 598 patients underwent thyroid surgery for CPT; there were 39 cases of TCC. However, no TCC were identified before 2006. The prevalence between 2008 and 2010 (36 patients) was 18.4%. Capsular invasion (82 % vs. 33%, p[lt]0.0001) and lymph node metastasis (35.8% vs 17.9% ,p [lt]0.002) were significantly more frequent in TCC than C patients at surgery. 196 patients that were operated for CPT between 2008 and 2010 were considered for the outcome assessment. All of them underwent a total thyroidectomy, 152 (77.5%) were ablated with I131 and 122 (62%) received TSH suppression with levothyroxine. With the stringent definition we used, no differences were found in the outcome of TCC (44.4% free of disease) and C (30%) at twelve months after surgery.[br]Conclusions: The prevalence of TCC was 18.4% of CPT operated in our hospital as from 2008. In spite of more aggressive pathological features at presentation, outcome at 12 months did not differ between TCC and C patients.[br][br]Nothing to Disclose: MFRP, CC, AJ, MF, EM, AM, AK, MB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 544 64 524 SAT-435 PO41-01 Saturday 463 2012


464 ENDO12L_SAT-436 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Prevalence of Thyroid Cancer in Minority Patients with Thyroid Nodules at an Urban Inner City Hospital Nazia Sultana, Zwadie March, Rhonda Trousdale Harlem Hospital, New York, NY; Columbia University Medical Center, New York, NY The National Cancer Institute has reported that non-Hispanic Whites and White Hispanics have a similar incidence of thyroid cancer (CA) (2.3 and 5.4 per 100,000 males and females per year)(1,2). In contrast Blacks have a significantly lower incidence of thyroid CA (1.3 and 3.7 per 100,00 black males and females per year) and only account for 5% of all cases of thyroid cancer.[br]One theory for the low incidence in Blacks is poor access to healthcare and lower screening rates leading to under-detection of thyroid CA. If the primary issue with incidence is screening, the prevalence of thyroid CA amongst patients with thyroid nodules should be similar for White, Hispanic and Black pts.[br]The prevalence of thyroid CA in pts with a nodule [gt]10 mm has been reported at 14.8% for a general population (GP)(3). The prevalence of thyroid CA in minorities with thyroid nodules has not been previously reported.[br]Objective: Determine the prevalence of thyroid CA in Black and Hispanic patients with a thyroid nodule [gt]10mm on US.[br]Setting: Urban, inner-city community hospital.[br]Design: Retrospective observational study of all patients referred for thyroid US between 2006-2011.[br]Results: 1,162 pts underwent thyroid US, 731 had nodules, 516 pts had at least one nodule [gt]10mm, 289 pts had FNA and/or surgical pathology report available. Ethnic distribution of pts with thyroid nodules (61% Black, 34% Hispanic, 4% Other) was not significantly different than the ethnic distribution of the neighborhood (48% Black, 35% Hispanic, 17% Other).[br]There were 17 new cases of thyroid CA identified (2 Black, 15 Hispanic). The prevalence of thyroid CA in Hispanics (15%) was equal to previously reported prevalence of 15% in GP(1). The prevalence of thyroid CA in Blacks (1.2%) was statistically lower than both Hispanics and GP(p[lt]0.001).[br]At Harlem Hospital, Hispanics with a thyroid nodule [gt]10mm have a 11.8 relative risk of thyroid CA compared to Black pts (95% CI 2.75-50.71;p[lt]0.0001).[br]Conclusion: The ethnic distribution of pts with thyroid nodules at an urban inner-city hospital closely reflected the ethnic make-up of the surrounding neighborhood. The prevalence of thyroid CA in Hispanics with a thyroid nodule [gt]10mm was similar to previously reported GP rates. However, the prevalence of thyroid CA in Blacks with a documented thyroid nodule was very low. This study demonstrates that the previously reported low incidence of thyroid CA in Black populations cannot be fully attributed to low screening rates.[br][br](1)Spitz MR et al., Int J Cancer 1988; 42:549. (2)Yu G-P et al., Thyroid 2010; 20:465. (3)Frates MC et al., J Clin Endocrinol Metab 2006; 91:3411.[br][br]Nothing to Disclose: NS, ZM, RT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 880 64 525 SAT-436 PO41-01 Saturday 464 2012


465 ENDO12L_SAT-437 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) A Profile of Thyroid Cancer in the West of Ireland Ruth Casey, Aoife Garrahy, Mary B Casey, Marcia Bell Galway University Hospital, Galway, Ireland; Galway University Hospital, Galway, Ireland Worldwide there has been a recent increase in the incidence of thyroid cancer (TC).[sup]1[/sup] Data on TC in Ireland is limited by the lack of an established TC register. The aim of this study was to profile TC in a single institution in the West of Ireland. A retrospective review of all histology specimens attained at fine needle aspiration (FNA) and thyroidectomy between 2008 and 2011 was carried out. Computer based radiology, pathology and biochemical data bases were interrogated for relevant information. A total of 151 patients, 124 females were included in the study. Mean age was 50 (SD 16.3). A total of 61 patients had malignant pathology. Papillary carcinoma was the most common subtype and accounted for 64%. The pathological diagnosis was made preoperatively using FNA in 48%. The remaining diagnoses were made from specimens post thyroidectomy. The most common indication for thyroidectomy in patients without a preceding FNA was suspicion of malignancy. Other indications from thyroidectomy included compressive effects of large goitres and management of hyperthyroidism. Youth was found to be a predictor of malignant pathology (p 0.002). Male gender approached statistical significance (p 0.07). Thyroid function tests were available for 98% of patients preoperatively. 75.5% were biochemically euthyroid. Thyroid status was not a predictor of malignancy (p 0.4). 72% underwent a preoperative ultrasound. Sonographic findings of a solitary thyroid nodule, dominant nodule within a multinodular goitre and a nodule size [gt]2cm correlated positively with malignancy (p 0.002). This highlights the usefulness of ultrasound in the investigation of thyroid disease. As expected papillary carcinoma was the most common subtype of TC [sup]1[/sup] in this Institution. Surprisingly in this population there was a higher percentage of male patients with TC than expected (25%). This study highlights the need for a dedicated TC register.[br][br](1) IARC; 2007. IARC Scientific Publications No. 160.[br][br]Nothing to Disclose: RC, AG, MBC, MB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2133 64 526 SAT-437 PO41-01 Saturday 465 2012


466 ENDO12L_SAT-438 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Changes in Prognosis and Risk Factors of Papillary Thyroid Cancer in Korea during Past Decades Hoon Sung Choi, Hwa Young Ahn, Jung Ah Lim, Sang Wan Kim, Young Joo Park, Ka Hee Yi, Do Joon Park, Seong Yeon Kim, Bo Youn Cho Seoul National University College of Medicine, Seoul, Republic of Korea; Seoul National University Bundang Hospital, Seongnam, Republic of Korea; Boramae Municipal Hospital, Seoul, Republic of Korea; Chung-Ang University College of Medicine, Seoul, Republic of Korea Backgrounds: The recurrence rates and mortality rates of papillary thyroid cancer (PTC) is reported to be reduced, with significant increase in micro-PTC. Rapidly increasing rates of PTC in Korea also indicates improvement of its prognosis, although only few studies were done. We examined the chronological changes in prognosis and associated risk factors of PTC patients in Korea.[br]Methods: Our study included 3147 patients who were diagnosed as PTC at the Thyroid Cancer Clinic in Seoul National University Hospital from 1962 to 2009. The clinicopathological characteristics and prognosis were collected by medical records review for analysis of changes in prognosis and risk factors.[br]Results: Patients[apos] mean age was 46.5[plusmn]12.6 years and female gender was 84.2%. During follow up periods of median 5.1 years (1-43), the cumulative recurrence rates were 18% at 10 year and 31% at 20 year, and the cumulative mortality rates were 1% at 10 year and 6% at 20 year. According to years at diagnosis (before 1989, from 1990 to 1999, after 2000), recurrence rates and mortality rates were significantly decreased with the passage of time. The associated factors for recurrence were male (Odds ratio [OR], 1.579; 95% confidence intervals [CI], 1.001-2.489; P=0.05), large size tumor (OR, 1.804; 95% CI, 1.564-2.081; P[lt]0.001), lymph node (LN) involvement (OR, 3.660; 95% CI, 2.177-6.153; P[lt]0.001), and extrathyroidal extension (ETE) (OR, 2.142; 95% CI, 1.161-3.953; P=0.015). Among them, OR of male was decreased, and OR of tumor size, LN involvement and ETE were increased as time passed. Regarding mortality, old age (OR, 17.021; 95% CI, 3.719-77.909; P[lt]0.001), male (OR, 4.204; 95% CI, 1.448-12.204; P=0.008), large size tumor (OR, 1.713; 95% CI, 1.289-2.277; P[lt]0.001), and LN involvement (OR, 10.584; 95% CI, 1.331-84.136; P=0.026) showed meaningful effects with the passage of time. The male gender seized to be a meaningful factor after 2000. In contrast, age and tumor size showed constant association regardless of time.[br]Conclusion: The prognosis of PTC in Korea, like the rest of the world, has improved and shows change in risk factors. Among the risk factors associated with recurrence, chorological characteristics showed decreasing association. In contrast, pathological showed more significant association. Regarding mortality, the male gender no longer showed meaningful association, while age and tumor size showed constant risk association.[br][br]Nothing to Disclose: HSC, HYA, JAL, SWK, YJP, KHY, DJP, SYK, BYC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 794 64 527 SAT-438 PO41-01 Saturday 466 2012


467 ENDO12L_SAT-439 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) BRAF Mutation in Follicular Variant of Papillary Thyroid Carcinoma Is Correlated with Suspicious Ultrasonographic Features without Clinicopathological Difference Dong Yeob Shin, Sena Hwang, Sooyun Chang, Eun Jig Lee Yonsei University College of Medicine, Seoul, Republic of Korea Background: Follicular variant of papillary thyroid carcinoma (FVPTC) is the most common subtype of papillary thyroid carcinoma. However, optimal classification and clinical behavior of FVPTC is debated. The objective of this study was the to investigate the correlation of BRAF V600E mutation with clinicopathological and ultrasonography (US) features in FVPTC.[br]Patients and methods: A total of 114 patients with pathologically confirmed FVPTC, who had BRAF V600E mutation analysis at the preoperative US-guided fine needle aspiration biopsy were included. Clinicopathlogical parameters and the frequency of suspicious US features were compared between BRAF V600E positive and negative groups. [br]Results: 48.3% of FVPTC patients were BRAF V600E mutated. Logistic regression showed that BRAF V600E mutation was not statistically correlated with age, multiplicity, tumor size, central/lateral cervical lymph node metastasis (LNM), and extra-thyroidal extension in FVPTC. However, BRAF V600E positive cases had a significantly higher possibility to present with both classical PTC and FVPTC subtypes (47.27% vs 20.34%, P=0.003) to have BRAF V600E mutation. FVPTC with BRAF V600E mutation also showed higher frequency of suspicious US features (90.7% vs 74.6%, P=0.025) and smaller tumor size on US (8.3 [plusmn] 0.4 mm vs 1.1 [plusmn] 0.9 mm, P=0.028).[br]Conclusions: In FVPTC, BRAF V600E mutation was correlated with higher frequency of suspicious US features despite smaller size on US. However, other clinicopathological parameters including LNM were not different according to BRAF status in FVPTC.[br][br]Nothing to Disclose: DYS, SH, SC, EJL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1395 64 528 SAT-439 PO41-01 Saturday 467 2012


468 ENDO12L_SAT-440 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Thyroid-Stimulating Hormone Level in Differentiated Thyroid Cancer Is Associated with Size and Advanced Stage of the Cancer Won-Chul Ha, Su-Jin Oh, Moo-Il Kang, Hyun-Shik Son, Tae-Seo Sohn, Ye-Jee Lim Uijeongbu St Mary[apos]s Hospital, Uijeongbu-si, Korea; Seoul St Mary[apos]s Hospital, Seoul, Korea The prevalence of thyroid cancer is increasing and recent studies have shown that serum thyroid-stimulating hormone (TSH) level is associated with an increased risk of thyroid cancers. The aim of this study is to evaluate association between TSH level and size and stage of thyroid cancer.[br]Method[br]The study was a retrospective cross-sectional analysis. Data from 199 patients with normal thyroid function underwent thyroid surgery for thyroid cancer between 2005 and 2011 was analyzed (81% female, age 52.2[plusmn]11.7 years). Thyroid cancers were diagnosed on final pathology and staged according to tumor node metastasis in all patients. The patients were subdivided into three groups based on normal mean TSH level predetermined in a population study (0.17[sim]0.4 mU/L, 0.4[sim]1.86 mU/L, 1.86[sim]4.65 mU/L).[br]Result[br]Of the 199 patients, 101(50.8%) patients were stage I[sim]II and 98(49.2%) patients were stage III[sim]IV. The TSH level was 1.90[plusmn]1.02 mU/L and the size of thyroid cancer was 1.29[plusmn]1.08 cm. There was a positive relation between serum TSH level and thyroid cancer size, analyzed by linear regression (P=0.018). The patients with advanced stage (III[sim]IV) had a higher TSH level and larger cancer size compared with those with stage I[sim]II (2.01[plusmn]1.04 mU/L [italic]vs[/italic]. 1.79[plusmn]0.98 mU/L, 1.76[plusmn]1.22 cm [italic]vs[/italic]. 0.83[plusmn]0.65cm, P=0.01). Moreover, the mean TSH level of the patients with the advanced stage (2.01[plusmn]1.04 mU/L) was higher than the mean TSH level predetermined in a population study (1.86 mU/L). More patients with the advanced stage (III[sim]IV) was in the upper one-third TSH level (1.86[lt]TSH[le]4.65: 52/98 (53.1%)) than the patients with early stage (I[sim]II) (35/101(34.7%)) (P[lt]0.05).[br]Conclusion[br]TSH level is correlated with the size and stage of the thyroid cancer. Higher TSH levels than population mean, even within normal range, have been associated with an advanced stage of thyroid cancer. TSH might play a role in the advance of thyroid cancer.[br][br]Nothing to Disclose: W-CH, S-JO, M-IK, H-SS, T-SS, Y-JL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1175 64 529 SAT-440 PO41-01 Saturday 468 2012


469 ENDO12L_SAT-441 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Infiltration of Mixture of Immune Cells Is Associated with Low Agressiveness and Molecular Profile of Differentiated Thyroid Cancer Lucas Leite Cunha, Elaine Cristina Morari, Ana Carolina Trindade Guilhen, Daniela Razolli, Suely Nonogaki, Fernando Augusto Soares, Jose Vassallo, Laura Sterian Ward University of Campinas (Unicamp), Campinas, Brazil; University of Campinas (Unicamp), Campinas, Brazil; Adolfo Lutz Institute, S[atilde]o Paulo, Brazil; A C Camargo Cancer Hospital, S[atilde]o Paulo, Brazil; University of Campinas (Unicamp), Campinas, Brazil The immune responses against differentiated thyroid carcinomas (DTC) have long been recognized but their effects on patients prognosis and association with tumor molecular profile remain unclear. We studied 398 nodules (253 papillary and 13 follicular thyroid cancers; 132 nonmalignant tissues). We defined tumors as persistent/recurrent and/or presenting long distance metastasis according to serum Tg levels and/or image evidences of metastasis. Three hundred and twenty-six patients evolved free-of-disease after 43.50[plusmn]33.29 months of follow-up. Immune cell infiltration was identified using CD3, CD4, CD8, CD20, CD68, and FoxP3 immunohistochemical markers. We colocalized CD4/IL-17 in order to identify Th17 lymphocytic infiltration and colocalized CD33/CD11b to identify myeloid derived suppressor cells (MDSCs) infiltration. DTC molecular profile was assessed by immunohistochemical identification of MUC1, NIS, ATM, PTEN and B7H1 proteins. We observed immune cells infiltrating malignant more often than benign lesions and a close correlation among concurrent chronic lymphocytic thyroiditis (CLT), tumor infiltrating lymphocytes, tumor-associated macrophages (TAM) and cellular molecular profile, suggesting this profile may be related to antitumor immune response. The presence concurrent CLT (p=0.0026), CD68+ (p=0.04963), CD4+ (p=0.0224), CD8+ (p=0.03119), CD20+ (p=0.0134), FoxP3+ (p=0.0122), Th17 (p=0.06417) was associated with clinical and pathological features of lower tumor aggressiveness and a more favorable patient outcome. A log-rank test confirmed an association between CLT (p=0.00149), TAM (p=0.04963), and CD8+ lymphocytes infiltration (p=0.03119) and an increased disease-free survival. Furthermore, expression of MUC1, NIS, ATM, PTEN was accompanied by infiltration of a mixture of different immune cells. Tumors with high B7H1 expression were frequently enriched with MDSCs (p=0.03256), suggesting that a complex immune evasion mechanism is engaged in DTC microenvironment. However, the close association between concurrent CLT, CD68+, CD4+, CD8+, CD20+, FoxP3+, Th17 and low aggressiveness features suggest that immune elimination overcomes immune escape, explaining the good prognosis frequently associated to DTC.[br][br]Sources of Research Support: Fapesp: 2009/18362-0.[br][br]Nothing to Disclose: LLC, ECM, ACTG, DR, SN, FAS, JV, LSW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 974 64 530 SAT-441 PO41-01 Saturday 469 2012


470 ENDO12L_SAT-442 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Unilateral Parathyroidectomy May Fail To Diagnose 75% of Clinically Significant Concomitant Thyroid Cancers Douglas Politz, Jose Lopez, James Norman Norman Parathyroid Center, Tampa, FL Background: It is possible that unilateral parathyroidectomy may be missing clinically important thyroid cancers that are occasionally found incidentally.[br]Methods: Thyroid pathology from 15,000 consecutive patients undergoing parathyroidectomy for pHPT was examined for extent of parathyroid operation, influence of preop ultrasound, and eventual surgical outcome. Patients with preoperatively known concomitant thyroid cancer were excluded.[br]Results: 3091 (20.6%) parathyroid operations were unilateral while 11,909 (79.4%) were bilateral. Ultrasound was performed prior to surgical referral on 10,667 (71%) patients, identifying one or more thyroid nodules [gt]1cm in 5,227 (49%). FNA thyroid biopsy was performed on 278 (5.3%) prior to surgery with 24 (7.9%) being positive for thyroid cancer and excluded from the study.[br]During parathyroidectomy 5,507 thyroid nodules were removed in 4382 patients (29.2%). Average nodule size was 1.4+0.6cm. More than one nodule was removed in 1127 patients, all in the bilateral group (p[lt]0.0001). Thyroid nodules were removed/biopsied in 343 (11.1%) unilateral operations and 4039 (33.9%) of bilateral (p[lt]0.0001). The rate of thyroid nodule removal was identical (p=0.89) for bilateral patients regardless of whether ultrasound was performed or not. In contrast, biopsy of a thyroid nodule occurred one third as frequently when a unilateral operation was performed without ultrasound, and one half as often if ultrasound was performed (both p[lt]0.001). Well differentiated thyroid cancer was found in 25 (0.8%) unilateral and 274 (2.3%) bilateral operations (p[lt]0.0001). Clinically significant cancers ([gt]1cm) were found in 3 (0.1%) unilateral and 71 (0.6%) of bilateral operations (p[lt]0.0001). Ultrasound did not increase the frequency of incidental cancer detection in either group (p=0.07).[br]Conclusions: Incidental thyroid cancer is present in 2.3% of patients undergoing parathyroidectomy, but only one-fourth (0.8%) are detected in patients undergoing unilateral operation including less than 75% of clinically significant thyroid cancers. Although preoperative ultrasound may detect concomitant thyroid cancer prior to surgery, it did not increase the detection of incidental cancers. Although patient selection for unilateral operation likely plays a significant role, unilateral parathyroidectomy will fail to diagnose the majority of clinically significant thyroid cancers.[br][br]Nothing to Disclose: DP, JL, JN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1490 64 531 SAT-442 PO41-01 Saturday 470 2012


471 ENDO12L_SAT-443 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Comparative Incidence of Thyroid Cancer in Multinodular Goiter vs. Single Nodule: A Systematic Review and Meta-Analysis Juan P Brito, Andres Yarur, Bryan McIver, Victor M Montori Mayo Clinic, Rochester, MN; Miller School of Medicine, University of Miami, Miami, FL Context:[br]The extent to which the incidence of thyroid cancer is different in thyroid glands with a solitary nodule (SN) vs. multinodular goiter (MNG) remains uncertain.[br]Objective:[br]To summarize the extant literature on the comparative incidence of thyroid cancer in SN compared with MNG.[br]Data source:[br]Systematic search of included PubMed MEDLINE, Ovid Medline In-Process [amp] Other Non-Indexed Citations, Ovid MEDLINE, Ovid EMBASE, Ovid Cochrane Database of Systematic Reviews, Ovid Cochrane Central Register of Controlled Trials, and Scopus (through July 2011) and reference lists from retrieved articles.[br]Study Selection:[br]Two reviewers working independently selected observational cross-sectional and longitudinal studies evaluating thyroid cancer in SN and MNG.[br]Data Extraction:[br]Two reviewers working independently extracted descriptive, methodological, and quantitative information from each study with consensus resolution of discrepancies. We used the Ottawa-Newcastle tool to assess for the risk of bias in these observational studies.[br]Data Analysis:[br]Using random-effects meta-analyses we estimated pooled odds ratio of thyroid cancer in SN vs. MNG and its 95% confidence interval. We also estimated the I2 statistic that quantifies the proportion of differences in estimates across studies that is not attributable to chance variation and thus quantifies true inconsistency across study results. We explored for subgroup interaction by gender, diagnostic method, iodine-sufficiency status and type of study.[br]Results:[br]Fourteen studies from 7 countries and studying 20723 SN and 23565 MNG patients were eligible for inclusion. Most eligible studies were of adequate methodological quality. Multinodular goiters were associated with a lower risk of thyroid cancer than SN (pooled odds ratio [OR] 0.8, 95% confidence interval [CI], 0.67, 0.96; I2 = 35%%). Subgroup analysis demonstrated that this difference in risk persisted only when studies were conducted in countries other than United States (OR 0.71;95% CI, 0.60-0.83; I2 =11%).[br]Conclusions:[br]Evidence of low to moderate quality suggests that thyroid cancer may be less frequent in MNG than in SN, particularly outside the US and perhaps in iodine deficient areas.[br][br]Nothing to Disclose: JPB, AY, BM, VMM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 651 64 532 SAT-443 PO41-01 Saturday 471 2012


472 ENDO12L_SAT-444 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Coexistence of Multiple Lipomatosis and Differentiated Thyroid Cancer: More Than a Coincidence? Beatriz Lecumberri, Jersy Cardenas, Sonia Rodado, Antonio Santiago, Blanca Vicandi, Joaquin Diaz, Jose Maria Viguer, Luis Felipe Pallardo La Paz University Hospital, Madrid, Spain; La Paz University Hospital, Madrid, Spain; La Paz University Hospital, Madrid, Spain; La Paz University Hospital, Madrid, Spain; La Paz University Hospital, Madrid, Spain Subcutaneous lipomas (SL) are the most common benign mesenchymal tumors with an estimated prevalence of 1%. They usually appear single, between 50-60 years of age, without gender differences. However, multiple lipomatosis (ML) occurs in 5-10% of patients, more frequently in men and almost 30% of them have family history of ML. Cytogenetic abnormalities are found in 50-80% of cases, including rearrangements of the 12q13-15 region, and aberrations at 6p21 also described in follicular thyroid adenomas and aggressive subtypes of thyroid carcinomas (1), and the presence of an underlying mitochondrial dysfunction has been suggested. To date, there are still no reports of SL/ML prevalence in patients with differentiated thyroid cancer (DTC). We have reviewed 245 patients with DTC, identified those with SL/ML and compared their features with those of the whole DTC group. Among 198 women and 47 men (19%) with DTC, being 13 H[uuml]rthle cell carcinomas (HCC) (5%), we found 21 patients, 13 women and 8 men (38%), with ML mainly in arms, back and legs. Seventeen of them (80%) referred appearance of ML in several members of their families at an early age. In the ML group there were 6 HCC (28%) (4 mixed with areas of multicentric papillary thyroid microcarcinoma), 1 multicentric mixed papillary-follicular carcinoma, and 14 papillary thyroid carcinomas (PTC) (8 follicular variant, 1 diffuse sclerosing variant, 4 microcarcinomas, 7 multicentric and 8 showing capsular invasion). Five men and 1 woman had suffered a non-thyroid cancer (NTC): 1 seminoma, 2 melanomas, 1 colon, 1 nasopharyngeal carcinoma and 1 acute lymphoblastic leukemia. Sixteen patients (76%) had first-degree relatives with NTC mostly breast, lung and liver. Only 4 cases (19%) remained free of DTC at the end of the follow-up in the ML group. When compared to the DTC group, ML was significantly associated with a higher rate of men, HCC, mixed forms (HCC-PTC), multicentricity and less probability of remaining free of disease (all p[lt]0,001), and a non-significant trend towards older age at DTC diagnosis and higher prevalence of capsular invasion was found. In our series the prevalence of ML (8,5%) is significantly higher than that of the general population with an increased frequency of familiar forms. Based on our findings coexistence of ML and DTC seems to be not casual, and could be considered a new bad prognostic marker in DTC. More research is needed to unravel the mechanisms that underlie this association.[br][br](1) Wreesmann et al., Cancer Res 2004; 64:3780-3789.[br][br]Nothing to Disclose: BL, JC, SR, AS, BV, JD, JMV, LFP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1120 64 533 SAT-444 PO41-01 Saturday 472 2012


473 ENDO12L_SAT-445 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Alcohol Abstinence Reduces the Risk of Thyroid Cancer in Patients with Thyroid Nodules Swaytha V Yalamanchi, Rebecca S Sippel, Herbert Chen University of Wisconsin, Madison, WI Background: Thyroid cancer is the most rapidly increasing malignancy in the USA. While the increased use of cervical ultrasound accounts for some of this rise, other environmental or behavioral factors are thought to play significant role. However, to date, few of these reasons have been clearly identified. In the study, we examined if alcohol consumption or tobacco use are risk factors for the development of thyroid cancer.[br]Methods: We retrospectively reviewed a prospective database of 1066 consecutive patients undergoing thyroidectomy for nodular thyroid disease between 1999- 2011. The final pathology was categorized as benign or malignant. Data were analyzed with Chi-square and ANOVA tests with SPSS.[br]Results: The mean age of the patients was 48.12 [plusmn] 15.89 years, and 78% were female. Of the 1066 patients, 472 underwent thyroid lobectomy while 579 had a total thyroidectomy. A history of tobacco abuse was not associated with a risk of thyroid cancer (p-value = 0.826). In contrast, individuals who had a history of alcohol use had a significantly higher incidence of thyroid malignancy (34% vs. 28%, p- value = 0.022). However, the risk of thyroid cancer was not dependent upon the absolute amount of alcohol consumed on a weekly basis.[br]Conclusions: In patients undergoing thyroidectomy for nodular thyroid disease, alcohol consumption is a significant risk factor for thyroid cancer. This observation may be due to alcohol or possibly due to other behavioral factor associated with alcohol abstinence.[br][br]Nothing to Disclose: SVY, RSS, HC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1568 64 534 SAT-445 PO41-01 Saturday 473 2012


474 ENDO12L_SAT-446 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Immunodetection of Cancer Stem Cell Markers in Anaplastic Thyroid Carcinoma Georges Elhomsy, Reigh-Yi Lin Saint Louis University, Saint Louis, MO [bold]Introduction:[/bold] Anaplastic thyroid carcinoma (ATC) accounts for 1-2% of all thyroid cancer, is a fatal disease, with a median survival rate of 6 months. This low survival rate is due to the aggressive nature of the disease and the lack of efficacious therapy. A better understanding of thyroid carcinogenesis is needed to improve the outcome of this cancer through novel therapies.[br][bold]Background: [/bold]It was hypothesized that only a small subset of cancer cells, referred to as cancer stem cells (CSC), have the ability to replicate indefinitely and produce diverse tumor cells that sustain tumor growth and metastasis. To evaluate a potential role of these CSC markers in ATC, we investigated the expression profiles of four putative CSC markers in tissues from ATC patients. These markers include NANOG, a transcription factor expressed in embryonic stem cells; studies showed that NANOG was also expressed in brain CSC. Chemokine receptor type 4 (CXCR4) a CSC marker in lung cancer. CD44, a cell-surface glycoprotein that plays a role in cell growth and differentiation, is a CSC marker in breast cancer and colorectal cancer. Finally, aldehyde dehydrogenase (ALDH), an enzyme plays a part in cellular detoxification, was proved to be a CSC marker in many cancers including lung cancer and breast cancer.[br][bold]Methods:[/bold] ten cases of ATC were examined. The expression of NANOG, CXCR4, CD44 and ALDH were analyzed in paraffin-embedded biospecimens by immunohistochemistry. Immunoreactivity of each CSC marker was scored by evaluating the number of positive tumor cells over the total number of tumor cells.[br][bold]Results:[/bold] The expression of all evaluated CSC markers differed significantly between patient samples. We found 6 of 10 subjects stained positive for CXCR4; 3 of 9 subjects stained positive for NANOG; 4 of 7 and 5 of 6 subjects stained positive for CD44 and ALDH respectively. In all markers, we found negative staining in normal thyroid cells adjacent to cancer cells. In addition, inter-individual variability in the disease was discovered. Only one patient was positive for all four markers, while two patients were positive for ALDH, CXCR4 and CD44 markers.[br][bold]Conclusion:[/bold] Our data suggest that CSC-associated pathways are activated in ATC. Because CSC is thought to be responsible for tumor initiation and its recurrence after chemotherapy, further large-scale studies are required to confirm these findings to establish the prognostic value of CSC in the clinical setting.[br][br]Nothing to Disclose: GE, R-YL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 156 64 535 SAT-446 PO41-01 Saturday 474 2012


475 ENDO12L_SAT-447 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) Different Alleles at Several [italic]Phosphodiesterase 8B (PDE8B)[/italic] Gene Loci Predispose to the Development of Either Secreting or Non-Secreting Thyroid Tumors Lionel Groussin, Laure Cazabat-Sage, Joel Coste, Fernande Rene-Corail, Diana Zelenika, Mark Lathrop, Pilar Galan, Jerome Clerc, Jerome Bertherat, Marie-Laure Raffin-Sanson Institut Cochin, Paris, France; Groupe Hospitalier Cochin-Saint Vincent de Paul Universit[eacute] Ren[eacute] Descartes Sorbonne Paris Cit[eacute], Paris, France; H[ocirc]pital Ambroise Par[eacute], Assistance Publique-H[ocirc]pitaux de Paris, Boulogne, France; Universit[eacute] Versailles-Saint Quentin en Yvelines, Paris-Saclay, Versailles, France; Groupe Hospitalier Cochin-Saint Vincent de Paul Universit[eacute] Ren[eacute] Descartes Sorbonne Paris Cit[eacute], Paris, France; Groupe Hospitalier Cochin-Saint Vincent de Paul Universit[eacute] Ren[eacute] Descartes Sorbonne Paris Cit[eacute], Paris, France; Centre d[apos]Etude du Polymorphisme Humain, [Eacute]vry, France; INRA/CNAM,Universit[eacute] Paris 13, Bobigny, France [bold][underline]Context[/underline][/bold]: Thyroid growth and secretion is controlled by TSH through activation of the of cyclic AMP pathway. Mutations activating this pathway result in hypersecreting thyroid adenomas. Higher TSH plasma levels, although remaining within the normal range, have been associated with a higher risk of thyroid tumors. Phosphodiesterase 8B (PDE8B) is strongly expressed in thyroid cells where it controls AMPc concentration and thus the activity of the AMPc pathway. Genotype at different snps in the [italic]PDE8B[/italic] gene have been associated with opposite TSHus and T4l plasma variations in several populations suggesting a different sensitivity to TSH in the carriers (1)(2).[br][bold][underline]Objective[/underline][/bold]: We hypothesized that alleles associated with higher TSH plasma levels could be more frequent in patients with thyroid tumors.[br][bold][underline]Design and Setting[/underline][/bold]: Four snps at the [italic]PDE8B[/italic] gene (rs4704397, rs6453293, rs4361497 and rs13158164) were genotyped in patients treated by surgery for different thyroid tumors and in control subjects with a normal thyroid palpation, all recruited in two University Hospital Departments.[br][bold][underline]Patients[/underline][/bold]: 247 patients with thyroid tumors (hypersecreting adenoma: n=21, benign non toxic multinodular goiter: n=93, papillary cancer: n=111, follicular cancer: n=22) and 313 matched controls. A complementary cohort of 2191 controls was also available for 2 snps.[br][bold][underline]Main Outcome Measure[/underline][/bold]: Allelic frequencies at the 4 loci.[br][bold][underline]Results[/underline][/bold]: We confirmed a higher frequency of the alleles associated with [underline]higher[/underline] TSH levels in patients with non toxic multinodular goiter (p=0,04) as well as in patients with papillary cancer (p=0,022). Interestingly, the small group of patients with hypersecreting thyroid tumors had a significantly higher frequency of the alleles associated with [underline]lower[/underline] TSH plasma levels in 3 out of the 4 snps.[br][bold][underline]Conclusion[/underline][/bold]: Depending on the genotype at four PDE8B loci associated with sensitivity to TSH in thyroid cells, subjects are at risk to develop either secreting or non secreting thyroid tumors.[br][br](1)Arnaud-Lopez L et al., Am J Hum Genet, 2008; 82(6): 1270[ndash]1280. (2)Taylor PN et al., Eur J Endocrinol. 2011; 164(5): 773[ndash]780.[br][br]Sources of Research Support: PHRC Regional, Assistance Publique H[ocirc]pitaux de Paris.[br][br]Nothing to Disclose: LG, LC-S, JC, FR-C, DZ, ML, PG, JC, JB, M-LR-S 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1477 64 536 SAT-447 PO41-01 Saturday 475 2012


476 ENDO12L_SAT-448 POSTER SESSION: Evaluation of Thyroid Nodules [amp] Prognosis/Risk Factors in Thyroid Cancer (1:30 PM-3:30 PM) IL-32 Promoter Polymorphism Modulates IL-32 Expression and Influences the Risk and the Outcome of Epithelial Cell Derived Thyroid Carcinoma Theo Plantinga, Irene Costantini, Bas Heinhuis, Angelique Huijbers, Benno Kusters, Mihai Netea, Ad Hermus, Charles Dinarello, Leo Joosten, Romana Netea-Maier Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands Objectives:[br]IL-32 is an intracellular proinflammatory cytokine that can modulate cell survival. Recent studies have suggested the involvement of IL-32 in the pathogenesis of malignancies. In the present study we aimed to elucidate whether genetic variation in the IL-32 promoter modulates IL-32 expression and whether it influences susceptibility and/or outcome of epithelial cell derived thyroid carcinoma (TC).[br]Methods:[br]Immunohistochemical staining for IL-32 was conducted on TC tumor tissue. Peripheral blood mononuclear cells from healthy donors with different genotypes for the IL-32 T/A promoter polymorphism rs28372698 were assessed for mRNA expression of IL-32 isoforms. Furthermore, a cohort of TC patients (N=139) was genotyped for this SNP and compared to a cohort of healthy controls (N=138).[br]Results:[br]IL-32 is highly expressed in TC tumor cells, compared to healthy tissue. Detection of mRNA expression of IL-32 in primary cells from healthy volunteers after LPS stimulation revealed two-fold higher expression of IL-32 in cells homozygous for the ancient T allele, compared to individuals homozygous for the variant A allele (p[lt]0.05). Furthermore, production of LPS-induced cytokines was increased in cells bearing the ancient T allele of the IL32 polymorphism. Genetic analysis for the IL32 promoter polymorphism revealed a different distribution of frequencies between TC patients and controls: the ancient T allele was overrepresented in TC patients, with an OR (95%CI) = 1.71 (1.06-2.75). Accordingly, the cumulative radioactive iodine dose required to eradicate the tumor after total thyroidectomy was significantly higher in TC patients bearing the ancient T allele.[br]Conclusions:[br]Individuals bearing genetic variants of IL32 that lead to increased IL-32 gene expression and a higher production of proinflammatory cytokines have an increased risk for developing epithelial cell derived TC. Subsequently, they require a higher dose of radioactive iodine to achieve successful tumor ablation. These data suggest an important role of IL-32 in the pathogenesis of TC.[br][br]Nothing to Disclose: TP, IC, BH, AH, BK, MN, AH, CD, LJ, RN-M 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1293 64 537 SAT-448 PO41-01 Saturday 476 2012


477 ENDO12L_SAT-449 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Neurofibromatosis Type 1 and Extra-Adrenal Composite Paraganglioma: An Unusual Association Amit Bhargava, Ria Lim, Sameera Tallapureddy, Socorro Vargas University of Connecticut Health Center, Farmington, CT; Saint Francis Hospital, Hartford, CT [bold]Background[/bold]. Composite tumors containing pheochromocytoma and ganglioneuroma are rare tumors typically arising in the adrenals. We present a case of extra-adrenal composite paraganglioma in a nonhypertensive patient with neurofibromatosis type 1 (NF1).[br][bold]Clinical case[/bold]. 34y normotensive female with NF1 and family history of NF1, on CT scan imaging for abdominal pain, was found to have a 6cm right-sided retroperitoneal mass. With her history of NF1, further testing for paraganglioma was done. Plasma free metanephrine was normal. Both plasma and 24hr urine normetanephrine were elevated at 7.03 nmol/L (reference[lt]0.9) and 2908 mcg/24hr (reference 111-419), respectively. Chromogranin A was also increased at 1675 ng/ml (reference[lt]225). I[sup]123[/sup] MIBG scan revealed intense uptake at the right upper quadrant of the abdomen. After alpha blockade with phenoxybenzamine, the 8x5x4cm retroperitoneal tumor was resected. Histology revealed cells consistent with pheochromocytoma mixed with foci showing neurofibrillary cytoplasmic processes [amp] ganglion cells representing ganglioneuroma. Immunohistochemical stains were positive for CD56 and chromogranin in the pheochromocytoma component and neurofilament immunoreactivity in the ganglioneuroma area. Post-operatively, plasma free normetanephrine and chromogranin A were normal.[br][bold]Discussion[/bold]. Reviewing literature, there is no reported case of NF1-associated composite paraganglioma outside the adrenals. In NF1, it is hypothesized that neurofibromin insufficiency may induce abnormal proliferation of Schwann cells and produce neurotropins that result in propagation of the pheochromocytoma and ganglionic cells, leading to composite pheochromocytomas. Clinical presentation is usually similar to pure pheochromocytomas. In some patients, there may be absence of endocrine abnormalities and symptoms possibly due to autoregulation of the pheochromocytoma by the ganglion cells of the adjacent ganglioneuroma components. Management is surgical resection, with preoperative alpha blockade. Grossly, it is similar in appearance to typical pheochromocytoma; hence the mixed phenotype is distinguished histologically. There is limited data on their malignancy potential. [br][bold]Conclusion[/bold]. Composite paraganglioma is a rare variant of an uncommon disease. As in this case, patients may not have the typical symptoms of pheochromocytoma. To our knowledge, there has been no reported case of NF1-associated extra-adrenal composite pheochromocytoma.[br][br]Nothing to Disclose: AB, RL, ST, SV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 759 65 538 SAT-449 PO45-01 Saturday 477 2012


478 ENDO12L_SAT-450 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) A Rare Case of a Benign Sporadic Pheochromocytoma Presenting with Reactive Hypoglycemia Prior to Subsequent Hypertension Deepa Beeharry, Rupa Ahluwalia, Alison Waghorn, Jiten Vora Royal Liverpool University Hospital, Liverpool, UK [underline]Case History:[/underline][br]A 65-year-old woman presented to our endocrinology unit with [apos]classical[apos] symptoms of post-prandial reactive hypoglycaemia. At the point of diagnosis, she was otherwise well with no o significant medical history of note.[br][underline]Investigations and method:[/underline][br]A 75gm glucose tolerance test (GTT) confirmed the diagnosis with a fasting glucose of 5.3 and a 2-hour reading of 1.8mmol/L. A normal 72-hour starvation test ruled out any evidence of unprovoked hypoglycaemia. Additional tests including baseline biochemistry, short synacthen test and thyroid hormone levels were normal. A barium enema revealed a small sigmoid diverticulum. During the course of investigations she was also found to be persistently hypertensive. Associated intermittent headaches with flushing led to further investigations for secondary causes of hypertension.[br][underline]Results and treatment:[/underline][br]Her 24-hour urinary collection showed markedly raised noradrenalin (2824 nmol/24 hrs; normal 120 to 590) and normetadrenaline (24684 nMol/24 hrs-normal [lt]650) levels. A CT scan of the abdomen revealed a 10cm left adrenal tumour. Functional scintigraphy (MIBG I123) was suggestive of a phaeochromocytoma. She later underwent laproscopic left adrenalectomy following preoperative alpha and beta blockade for blood pressure control. Histology confirmed a benign phaeochromocytoma.[br][underline]Conclusions and points for discussion:[/underline][br]Phaeochromocytomas are classically associated with hyperglycaemia due to catecholamine excess resulting in a combination of impaired insulin secretion primarily due to alpha-adrenoceptor stimulation and enhanced gluconeogenesis and glycogenolysis. Post-operative hypoglycaemia is a well-recognized feature of phaeochromocytomas as well. Loss of inhibitory effect of catecholamine excess resulting in rebound insulin production can cause hypoglycaemia following surgery.[br]Non-post- operative hypo glycaemia is a very rare complication of phaeochromocytoma, with only a limited number of reported cases in the literature. The underlying mechanism remains unclear and less well understood. Various possible mechanisms have been proposed including beta-adrenoceptor mediated insulin release as well as direct tumoural secretion of insulin or insulin like growth factors.[br]Our case highlights the importance of recognsing both hypoglycaemia as well as hyperglycaemia as associated features with phaeochromocytomas.[br][br]Nothing to Disclose: DB, RA, AW, JV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1412 65 539 SAT-450 PO45-01 Saturday 478 2012


479 ENDO12L_SAT-451 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Unusually Aggressive Paraganglioma Phenotype in a Kindred with a 57delG [italic]SDHD[/italic] Mutation Ria Lim, Jennifer Stroop, Faripour Forouhar, Robert Frankenthaler, Carl Malchoff University of Connecticut Health Center, Farmington, CT; University of Connecticut Health Center, Farmington, CT; University of Connecticut Health Center, Farmington, CT; Beth Israel Deaconess Medical Center, Boston, MA [bold]Background.[/bold] Familial paraganglioma syndromes (FPS) are caused by germline mutations in succinate dehydrogenase (SDH) subunit genes. Paragangliomas caused by [italic]SDHD[/italic] mutations are usually less aggressive than those caused by [italic]SDHB[/italic] mutations. In contrast, we describe three patients in a single kindred with a [italic]SDHD[/italic] mutation and unusually aggressive paragangliomas.[br][bold]Clinical Cases.[/bold] The index subject is a white male who presented at age 24 y with bilateral carotid body tumors. Debulking was followed by external beam radiation. Whole blood DNA sequencing identified a heterozygous 57delG [italic]SDHD[/italic] mutation. At age 31 y, he was found to have lung metastases that stabilized after treatment with Gemcitabine, high dose In-111-Octreotide, 100 mCi I-131-MIBG and 4 cycles of cytoxan, vincristine, and dacarbazine. At the age of 34 y bilateral pheochromocytomas were found. The sister of the index subject presented with bilateral carotid body tumors and a left glomus vagale tumor extending to the skull base at age 38 y. She underwent embolization of the glomus tumor, followed by resection of the glomus tumor and both carotid body tumors. The right carotid body tumor showed low mitotic activity, but demonstrated an infiltrative border composed of ribbon like cords invading the surrounding connective tissue. Whole blood DNA sequencing confirmed heterozygosity for the 57delG [italic]SDHD[/italic] mutation. Two sons of the index subject were heterozygous for the 57delG [italic]SDHD[/italic] mutation, and one developed a pheochromocytoma at age 8 y. The paternal grandmother of the index subject had bilateral paragangliomas, and the index subject[apos]s father was clinically unaffected.[br][bold]Discussion.[/bold] Paragangliomas caused by germline [italic]SDHD[/italic] mutations are less frequently malignant and occur at an older age than paragangliomas caused by germline [italic]SDHB[/italic] mutations. In paragangliomas caused by germline [italic]SDHD[/italic] mutations, the malignancy prevalence is estimated at 0-7%, and the age at onset ranges from 10-96 y. This kindred with a germline [italic]SDHD[/italic] mutation was unusual in that the paragangliomas were aggressive with metastases, infiltrative disease, and early onset. This suggests that the 57delG [italic]SDHD[/italic] mutation causes a more aggressive phenotype or that modifier genes in this kindred altered the expected phenotype. The expected imprinting persisted.[br][bold]Conclusion.[/bold] The paragangliomas in this kindred with the 57delG [italic]SDHD[/italic] mutation demonstrate a more aggressive phenotype than is usually seen in FPS caused by [italic]SDHD[/italic] mutations.[br][br]Nothing to Disclose: RL, JS, FF, RF, CM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 597 65 540 SAT-451 PO45-01 Saturday 479 2012


480 ENDO12L_SAT-452 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Localization of Occult Adrenal Tissue with Cosyntropin-Stimulated 18F-FDG-PET/CT in a Patient with Metachronous Adrenocortical Tumor Who Presented with Persistently Elevated DHEAS after Bilateral Adrenalectomy Maira Rovigatti Franco, Guilherme Asmar Alencar, Andre M Faria, Joao Evangelhista Bezerra, Carolina Cruz, Sorahia Domenice, Lorena Oliveira Lima, Antonio Marcondes Lerario, Madson Queiroz Almeida, Ana Claudia Latronico, Berenice Bilharinho Mendonca, Maria Candida Barisson Vilares Fragoso Hospital das Clinicas da Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Instituto do Cancer do Estado de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Instituto do Cancer do Estado de S[atilde]o Paulo, S[atilde]o Paulo, Brazil [bold]Introduction:[/bold] Adrenocortical carcinoma (ACC) is a rare and potentially fatal disease in childhood. Complete resection of the tumor and metastasis can improve survival. Integrated 2-[fluorine 18] fluoro-2-deoxy-D-glucose positron emission tomography/computed tomography ([sup]18[/sup]F-FDG-PET/CT) is a diagnostic method that has been used to discriminate benign from malignant adrenal masses (1). [bold]Case:[/bold] A 2.6 year-old boy presented isosexual pseudo-precocious puberty at 2.1 years. CT revealed a single nodule in the left adrenal (2 cm) which was resected by adrenalectomy. Histology revealed an adrenocortical tumor (2 cm, 3.0 g, Weiss score 4), ENSAT stage I. Partial regression of the puberty features and normalization of hormonal levels were observed. At 4.8 years, recurrence was observed, with high androgen levels and an 8 cm right adrenal mass on CT. Right adrenalectomy and nephrectomy was performed, which revealed an adrenocortical tumor (8 cm, 30 g, Weiss score 7) with no apparent metastasis at that time. At 6 years, DHEAS levels became increasingly high and CT showed a pulmonary nodule that was surgically removed. After 8 months of partial lung resection, abnormally high DHEAS was present with no obvious source on conventional imaging techniques. [bold]18F-FDG-PET/CT [/bold]showed a mediastinal lesion that was resected and histologically confirmed as metastatic ACC. DHEAS levels remained undetectable until he was 13 years-old, with a progressive increase not suppressible by high doses of dexamethasone (0.5 mg qid for 7 days). Conventional radiological techniques did not identify any metastatic lesions. At 18.5 years of age, the patient underwent 18F-FDG-PET/CT under stimulation with intravenous cosyntropin 250 mcg, which identified a 1.5 cm nodular lesion in left adrenal topography. FDG hypermetabolism was shown in that area pre-stimulus (SUV=1.5), with an increase 60 minutes after stimulation to 3.6, suggesting remaining tumorous tissue. [bold]Conclusions:[/bold] We report a long-term evolution of a pediatric patient with an indolent ACC and suspected local recurrence identified by cosyntropin-stimulated 18F-FDG-PET/CT. This new diagnostic modality may be used in clinical practice to locate occult metastasis of ACC.[br][br](1)Anurag R. Lila, Gaurav Malhotra et al.,Localization of Remnant and Ectopic Adrenal Tissues with Cosyntropin-Stimulated 18F-FDG-PET/CT in a Patient with Nelson Syndrome with Persistent Hypercortisolism. J Clin Endocrinol Metab, December 2010, 95(12):5172[ndash]5173.[br][br]Nothing to Disclose: MRF, GAA, AMF, JEB, CC, SD, LOL, AML, MQA, ACL, BBM, MCBVF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1610 65 541 SAT-452 PO45-01 Saturday 480 2012


481 ENDO12L_SAT-453 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Dopamine-Secreting Pheochromocytoma: A Different View of Catecholamine-Producing Tumor Ongkarn Sarasombath, James R Sowers, Guido Lastra University of Missouri, Columbia, MO Pheochromocytoma is a catecholamine producing tumor originating in adrenal medulla. The majority of these tumors secrete epinephrine (E) or norepinephrine (NE). Dopamine (DA) producing tumors are rare and usually do not present with classical symptoms of catecholamine excess.[br]Case: 43 year-old female in whom a CT abdomen was obtained during work up for right inguinal pain, and revealed a 5.8 x 3.3 cm heterogeneously enhancing mass in her left adrenal, which was confirmed by MRI. 24 hour urine dopamine was markedly elevated (28 times above the upper limit of normal). Plasma and 24 hour urine metanephrine and normetanephrine were mildly elevated (1.2 times). Plasma aldosterone concentration, plasma renin activity and dexamethasone suppression test were normal. With a diagnosis of a DA-secreting pheochromocytoma, the patient underwent left adrenalectomy. Surgery was well tolerated. Pathology report confirmed a left adrenal pheochromocytoma. The tumor cells were positive for chromogranin, synaptophysin, and neuron special enolase. Pheochromocytoma of the Adrenal gland Scaled Score (PASS) was 4 which imply a potential for a biologically aggressive behavior.[br]Discussion: Only fifteen cases of DA-secreting pheochromocytoma/paragangliomas have been reported in literature. These tumors lack the classical symptoms of E/NE secreting tumors, which can delay or obscure the diagnosis. The malignancy rate and incidence of extra-adrenal location is higher relative to E/NE producing tumors. The most reported diagnostic tests used are urinary and plasma dopamine. However, their sensitivities and specificities remain unknown. This type of tumor is less likely to be enhanced with MIBG compared to E/NE, and FDG-PET scan is another imaging option, its diagnostic accuracy is also unknown.[br]There are no specific guidelines for treatment of DA-secreting pheochromocytomas, although surgical resection is indicated. Unlike E/NE secreting tumors, preoperative [alpha]-blockade is not routinely recommended due to its potential for arterial hypotension.[br]This case points out the importance of identifying the type of tumor in a patient with adrenal mass. Knowing the nature of tumor will help guide the management. Measurement of urinary or plasma dopamine are part of workup in all patients with presumed pheochromocytoma.[br][br]Nothing to Disclose: OS, JRS, GL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 942 65 542 SAT-453 PO45-01 Saturday 481 2012


482 ENDO12L_SAT-454 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Vertebral Metastasis in a Case of New SDHB Mutation Eva Lau, Ana Varela, Ana Isabel Oliveira, Rui Pinto, Jorge Lima, Luis Matos Lima, Davide Carvalho Centro Hospitalar S[atilde]o Jo[atilde]o, Faculty of Medicine University of Porto, Porto, Portugal; Centro Hospitalar S[atilde]o Jo[atilde]o, Faculty of Medicine University of Porto, Porto, Portugal; Faculty of Medicine University of Porto, Porto, Portugal; Centro Hospitalar S[atilde]o Jo[atilde]o, Faculty of Medicine University of Porto, Porto, Portugal [bold]Introduction:[/bold] Paragangliomas are rare tumors that arise from nonadrenal chromaffin tissue. They are mostly benign. [bold]Case report[/bold]: 37 years-old Caucasian female, without known diseases until 1997, when she complains of headaches, restless feeling, anxiety and palpitations. Routine screening showed hypertension that was resistant to medical treatment. Laboratory analyses demonstrated an elevation of vanilmandelic acid (VMA 52 mg/24h (1.4-6.5)); epinephrine 21[micro]g/24h (0-20): norepinephrine 3161[micro]g/24h (23-105); dopamine 711[micro]g/24h (65-400); metanephrine 91[micro]g/24h (52-341) normetanephrine 3546 [micro]g/24h (88-444). Abdominal CT showed a 5.2 cm mass in the region of the left adrenal gland, with a corresponding hyperfixation in the MIBG scintigraphy. Intraoperatively it was found that the lesion was separated from the adrenal gland. This finding together with the pathologic report of a neoplasia invading the capsule, without going beyond it, and images of vascular venous invasion made the diagnosis of malignant paraganglioma. Genetic screening of the patient demonstrated a new germinal frameshift mutation of the SDHB exon 6 (c.587-591DelC). There was a family history of a cousin diagnosed at 13 years of age with a dopamine producing malignant paraganglioma, that died at 30 years of age with wide spread metastasis (cranial and lumbar). Genetic screening revealed: the father, two other cousins and one uncle had the mutation, till now without biochemical and morphological manifestations. The patient underwent more 3 surgeries. One to remove para-aortic metastasis/paraganglioma; the 2[sup]nd[/sup] to remove metastasis in the 11[sup]th[/sup] dorsal vertebral body (D11) and the last one to remove abdominal lymph node metastasis. Persistent elevation of catecholamines and persistent uptake of MIBG on D11 after surgeries led to [sup]131-[/sup]I-MIBG treatment twice with. The patient went also through radiation therapy (total dose of 30 Gy in 10 fractions) achieving stable disease. Two years after, urine and plasma fractionated catecholamines and metanephrines remain within normal range, with faint uptake of MIBG on D11, reflecting partial response. [bold]Conclusion:[/bold] This case outlines the clinical course of a patient with a familial disorder associated with paraganglioma syndrome type 4 caused by a new mutation in the SDHB gene. It also illustrates the importance of identifying patients with SHDB mutations, as these patients are at high risk of developing malignant disease.[br][br]Nothing to Disclose: EL, AV, AIO, RP, JL, LML, DC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1309 65 543 SAT-454 PO45-01 Saturday 482 2012


483 ENDO12L_SAT-455 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Pheochromocytoma: A Silent Killer Hassan Shawa Baylor College of Medicine, Houston, TX [bold]Background:[/bold] Clinical expression of pheochromocytoma may involve numerous cardiovascular manifestations due to catecholamine excess; some of them can be life-threatening such as ventricular arrhythmias, conduction disturbances, cardiogenic shock, or hypertensive emergencies. Cardiomyopathy is usually transient and reversible with treatment (1,2). Hypertension could be sustained or paroxysmal but approximately 10% of the cases are normotensive.[br][bold]Clinical Case[/bold]: A 38-year African-American woman presented with cardiogenic shock (EF [lt]15 %) that was unresponsive to placement of an intra-aortic balloon pump. She had a cardiac arrest and required cardiopulmonary resuscitation for 5 minutes. A TandemHeart was inserted emergently, and this allowed pressors to be weaned within minutes. Comprehensive electrophysiology with induction studies showed multifocal atrial tachycardia and junctional ectopic tachycardia. This was treated with radiofrequency ablation and a temporary pacemaker. One week later, another 2D-Echo showed an EF of [gt]70% and the TandemHeart was removed. CXR showed substantial pneumoperitoneum which was evaluated with a CT scan of the abdomen/pelvis. A 4.2 x 4.8 cm hypodense (14.8 HU) right suprarenal mass was noted. Plasma free fractionated metanephrines (metanephrine 1361 pg/ml, n[lt]57 pg/ml and normetanephrine 981 pg/ml, n[lt]148 pg/ml), Chromogranin A (19.0 ng/ml, n 1.9-15 ng/ml), 24hr urinary fractionated metanephrines (metanephrine 4749 ug/24 hr, n 36-190 ug/24 hr- normetanephrine 1434 ug/24 hr, n 35-482 ug/24 hr), catecholamines (epinephrine 222 ug/24 hr, n 2-24 ug/24 hr- norepinephrine 143 ug/24 hr, n 15-100 ug/24 hr), and VMA (10.8 mg/24 hr, n[lt]6 mg/24 hr) were elevated. Increased uptake in the right suprarenal mass only was confirmed by MIBG. BP was maintained in the normal range during while time. Salt and oral fluid intake were encouraged, and she was prepared for surgery with prazocin and subsequently carvedilol for 12 days. Doses were titrated up gradually without significant side effects. She underwent an open right adrenalectomy. Intraoperatively, her SBP went up to 210 mmHg requiring nitroprusside and Phentolamine transiently. Pathology confirmed the diagnosis of pheochromocytoma with no lymphovascular invasion.[br][bold]Conclusion:[/bold] Pheochromocytoma should be kept in the differential diagnosis while approaching young patients with acute cardiac decompensation even if they are normotensive and have no previous history of hypertension.[br][br]1. Wood R, Commerford PJ, Rose AG, Tooke A. Reversible catecholamine-induced cardiomyopathy. Am Heart J. 1991;121:610[ndash]613. 2. Bybee KA, Prasad A. Stress-related cardiomyopathy syndromes. Circulation. 2008;118:397[ndash]409.[br][br]Nothing to Disclose: HS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 620 65 544 SAT-455 PO45-01 Saturday 483 2012


484 ENDO12L_SAT-456 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Giant Biochemically Active Asymptomatic Pheochromocytoma Jonathan Stringer, Nabil Bissada, Vitaly Kantorovich University of Arkansas for Medical Sciences and Little Rock Veterans Affairs Hospital, Little Rock, AR; University of Arkansas for Medical Sciences and Little Rock Veterans Affairs Hospital, Little Rock, AR Pheochromocytoma (PHEO) is a rare tumor of chromaffin cells with potentially devastating clinical course. Its diagnosis may be delayed by a multitude of presentations, ranging from asymptomatic cases to severe PHEO crisis. Diagnosis relies on a timely clinical suspicion and assessment of either plasma or urine metanephrine levels. Surgery is the ultimate treatment, but patients need to be carefully prepared to prevent severe intra-operative complications. Here we present an unusual case of a giant asymptomatic PHEO. A 61 year old male was found to have a non-homogenous 12.8 x 7.6 x 8.0 cm adrenal incidentaloma on CT scan. Biochemical workup revealed plasma epinephrine of 256 (0-62 pg/ml), norepinephrine 1140 (0-874 pg/ml), metanephrine 540 (0-62 pg/ml) and normetanephrine [gt]20,000 (0-145 pg/ml), urine metanephrine 2075 (35-460 ug/24h) and normetanephrine 80,568 (110-1050 ug/24h). MIBG scan showed increased uptake in the area of the adrenal mass. Patient had a 12 years h/o well controlled hypertension on relatively low dose of atenolol and nifedipine. He was otherwise asymptomatic and his pre-operative echocardiography showed normal ejection fraction and no ventricular hypertrophy. He received phenoxbenazamine 2 weeks prior to surgery and was had mild orthostatic hypotension on 20mg BID. His post-operative course was uneventful with neither postural nor recumbent hypotension. Pathology revealed extension to local lymph nodes and Ki-67 of less than 1%. Plasma metanephrines were normal one month after surgery. Discussion: Several points can be raised by this very unusual case. Large PHEOs are thought to have lower synthetic capacity and together with easily controlled HTN, lack of hypertensive spells and normal ECHO could suggest a relatively benign disease with relatively low levels of catecholamines. This statement could also support low Ki-67. On the other hand, severely elevated intracellular metabolites [ndash] metanephrines [ndash] suggest both high synthetic and metabolic activity of the tumor. Chronic use of nifedipine might to some extent prevent vesicular degranulation, which diverted intracellular catecholamines towards metabolism to metanephrines rather than systemic spill over and, to some extent, precluded the development of systemic signs and end organ damage. Additionally, slow growth of the tumor could have been associated with marked desensitization of adrenergic receptors that explained lack of symptoms/signs, or of end organ damage.[br][br]Nothing to Disclose: JS, NB, VK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 275 65 545 SAT-456 PO45-01 Saturday 484 2012


485 ENDO12L_SAT-457 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Pheochromocytoma Presenting as Intestinal Pseudo-Obstruction: Case Report and Review of the Literature Melina Silva Pinto, Adriana Martins Fernandes, Daniel Soares Freire, Maria Adelaide Albergaria Pereira Hospital das Cl[iacute]nicas da Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Pheochromocytoma (pheo) is a rare catecholamine-secreting tumor originated from chromaffin cells. Intestinal pseudo-obstruction is a rare complication of the disease.[br]A 27 year-old man came for medical attention with a 4-day history of palpitation, cold sweating, postural hypotension, fever, dyspnea and dry cough. Chest x-ray revealed a wedge opacity on left lung. A chest CT scan identified a 11 cm tumor with necrotic center on the right adrenal gland. The patient was then referred to us.[br]On arrival, his blood pressure (BP) and heart rate (HR) at supine position were 197/135 mmHg and 117 bpm, respectively. In standing position, BP and HR were 140/33 mmHg and 148 bpm. The abdomen was moderately distended and tympanic. Bowell sounds were hypoactive and a rectal examination was normal.[br]The 24-hour urine samples demonstrated the following: total metanephrines, 14.0 (0.05-1.2 [mu]g/mg creatinine); norepinephrine, 5187 [mu]g/d (14-80 [mu]g/d); epinephrine, 729 [mu]g/d (0.5-20 [mu]g/d); dopamine, 490 [mu]g/d (65-400 [mu]g/d). Blood samples confirmed the urinary results: plasma norepinephrine 27789 pg/mL (40-268 pg/mL); epinephrine 1260 pg/mL (0-75 pg/mL); dopamine, undetectable (0-83 pg/mL).[br]Aggressive fluid replacement and treatment with prazosin were promptly started. Two days after admission the patient developed worsening of abdominal symptoms, with diffuse abdominal pain and distension, nausea and vomiting. Oral feeding was discontinued and a nasogastric tube was inserted, without complete resolution. The patient underwent an exploratory laparotomy, which found no anatomic obstruction, thus confirming the diagnosis of an intestinal pseudo-obstruction.[br]Since abdominal symptoms persisted despite alpha-blockage, prolonged parenteral nutrition was initiated while the patient was being prepared for surgery. One week after adrenalectomy, his bowel movements recovered and oral diet was restarted.[br]Intestinal pseudo-obstruction is one of the rarest manifestations of a pheochromocytoma. The underlying mechanism is related to the visceral effects of catecholamines acting via [alpha]-adrenergic receptors, which results in decreased motility, peristalsis and intestinal tone. Some authors have reported prompt relieve of symptoms after treatment with phentolamine, an [alpha][sub]1[/sub]/[alpha][sub]2[/sub]-adrenergic antagonist. Our patient was treated with prazosin, since phentolamine is not available in Brazil, with adequate blood pressure control but without any improvement of paralytic ileus, which resolved completely after surgery.[br][br]Nothing to Disclose: MSP, AMF, DSF, MAAP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1413 65 546 SAT-457 PO45-01 Saturday 485 2012


486 ENDO12L_SAT-458 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Catecholamine-Induced Myocarditis in Pheochromocytoma Valeria de Miguel, Andrea Paissan, Maria Arzadun, Anibal Arias, Diego Perez de Arenaza, Ariela Soto, Marcelo Pietrani, Patricia Fainstein Day Hospital Italiano de Buenos Aires, Buenos Aires, Argentina; Hospital Italiano de Buenos Aires, Buenos Aires, Argentina; Hospital Italiano de Buenos Aires, Buenos Aires, Argentina; Hospital Italiano de Buenos Aires, Buenos Aires, Argentina Background:[br]Pheochromocytomas are rare neuroendocrine tumors with variable clinical presentation. High circulating levels of catecholamines may cause direct myocardial injury.[br]Clinical case:[br]A 25-year old male patient arrived at the emergency room of his community hospital complaining of abdominal pain, headaches and palpitations while he was playing soccer. He had hypertension and the abdominal ultrasound revealed a tumor located in the right adrenal gland.[br]The patient was subsequently transferred to our hospital for further evaluation. Upon admission, he had an episode of acute pulmonary edema with severe hypertension (Blood pressure: 220/120 mm Hg). He was admitted to the Coronary Care Unit. Intravenous vasodilators and loop diuretics were administered with rapid recovery of clinical status.[br]Cardiac biomarkers: Creatinine-kinase (CK): 1145 UI/L (38-175), CK-MB: 115.9 UI/L ([lt]20), Troponin T: 1.38 ng/dl ([lt]0.04), pro BNP 1900 pg/dl ([lt] 450).[br]The ECG showed sinus rhythm, 80 beats per minute and T-wave inversion in DI, DII and AVL leads. Transthoracic echocardiography revealed left ventricular hypertrophy (LVH) and dilated left atrium. The ejection fraction was preserved.[br]Computed Tomography (CT) scan of the abdomen showed a 47.5 x 36.7 mm heterogeneous right adrenal mass.[br]The 24-hours urine fractionated metanephrines and catecholamines were elevated: metanephrine: [gt]1000 ug/24 h (25-115), normetanephrine: [gt]1000 ug/24 h (75-375), adrenaline: 52 ug/24 h (0.5-8.5), noradrenaline: 520 ug/24 h (18.5-100), vainillyl- mandelic acid: 9.9 mg/24 hs (0.2-8).[br]Cardiac Magnetic Resonance (CMR) showed increased left ventricular wall thickness and diffuse intramyocardial edema with focal intramyocardial late gadolinium enhancement (LGE).[br]After seven days of alpha- and beta- adrenergic receptor blockade, hemodynamic stability, the patient underwent a successful right laparoscopic adrenalectomy. Pathological studies confirmed the pheochromocytoma diagnosis.[br]Four-months after surgery he was asymptomatic, normotensive and urine catecholamines were within normal range. CMR depicted normal left ventricular wall thickness and reversal of myocardial edema with persistent focal LGE.[br]Conclusion:[br]Catecholamine induced- myocarditis is an infrequent clinical manifestation in patients with pheochromocytoma and should be distinguish from stress related cardiomyopathies.[br]It is important to recognize this pathology before surgical treatment in order to minimize morbidity and mortality.[br][br]Nothing to Disclose: VdM, AP, MA, AA, DPdA, AS, MP, PFD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 186 65 547 SAT-458 PO45-01 Saturday 486 2012


487 ENDO12L_SAT-459 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Pheochromocytoma during Pregnancy Zeina Maani, Srilakshmi Mitta, Peter Mazzaglia, Geetha Gopalakrishnan Brown University/Rhode Island Hospital, Providence, RI; Brown University/Rhode Island Hospital, Providence, RI; Columbia University, New York, NY Background: Pheochromocytoma during pregnancy is extremely rare. Although the severity and clinical features can vary, it can be a life-threatening condition for both the mother and the fetus. Prognosis has improved over the last decade due to a multidisciplinary approach in management. Regardless, timing and type of adrenal surgery as well as mode of delivery are debated. Even though vaginal delivery and laparoscopic adrenalectomy are less invasive, our case illustrates the need to consider c-section and open adrenalectomy in the management of large pheochromocytomas presenting in the third trimester.[br]Clinical case: A 34 year old G6P2032 female was referred for an 8 cm adrenal mass found on ultrasound at 23 weeks gestation. She denied any history of hypertension or family history of adrenal, calcium or thyroid problems. At the time of presentation, she was asymptomatic except for mild headaches. BP was 110/70 mmHg and HR was 68 bpm. Ultrasound revealed 8.1 x 5.2 x 7.5 cm heterogenous, solid right suprarenal mass with central echogenic and cystic areas. A 24 hour urine collection showed an elevated normetanephrine of 9042 mcg (nl [lt]900) consistent with a pheochromocytoma. Normal calcium, PTH and calcitonin levels were noted. She was treated with phenoxybenzamine and maintained normal BP and HR during pregnancy. However, at 39 weeks gestation, labor was induced and the patient developed severe hypertension. BP was controlled and a healthy baby girl was delivered via cesarean section. The patient was continued on phenozybenzamine. Laparoscopic adrenal surgery was scheduled 7 weeks later but was complicated by increased blood loss and renal ischemia requiring conversion to open surgery and right nephrectomy. Surgical pathology confirmed a 7x 7x 4.5 cm pheochromocytoma with focal necrosis and calcifications. Post-operatively, phenozybenzamine was discontinued. Follow-up blood pressure, heart rate, normetanephrine and metanephrines values were normal.[br]Conclusion: We present a case of pheochromocytoma during pregnancy that was diagnosed in the third trimester. Although vaginal delivery and laparoscopic adrenalectomy was considered in our patient, c-section and open adrenalectomy was eventually preformed due to complications. This case illustrates the benefits of c-section and open adrenalectomy when dealing with a large pheochromocytoma presenting during the third trimester.[br][br]Nothing to Disclose: ZM, SM, PM, GG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1263 65 548 SAT-459 PO45-01 Saturday 487 2012


488 ENDO12L_SAT-460 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Coexistence of Bilateral Myelolipoma and ACTH-Independent Macronodular Adrenal Hyperplasia with GIP/LH-Dependent Cushing Syndrome Louis Bondaz, Mathieu Latour, Kevin Zorn, Odile Prosmanne, Sylvie Oble, Livia Mara Mermejo, Andre Lacroix Centre Hospitalier de l[apos]Universit[eacute] de Montr[eacute]al (CHUM), Montr[eacute]al, Canada; CHUM, Montr[eacute]al, Canada; CHUM, Montr[eacute]al, Canada; CHUM, Montr[eacute]al, Canada [bold]Background:[/bold] ACTH-independent bilateral macronodular adrenal hyperplasia (AIMAH) is a rare cause of Cushing[apos]s syndrome, but is frequently secondary to the aberrant expression of various G-protein coupled hormone receptors. Adrenal myelolipoma is a rare benign neoplasm composed of mature adipose tissue and variable amount of hematopoietic elements. Its pathogenesis remains unclear. It is usually unilateral and not associated with hormonal secretion, but has rarely been described in association with inflammatory conditions, or with hyperfunctioning adrenal disorders. We now report its presence in a case of AIMAH with GIP/LH dependent Cushing[apos]s syndrome.[br][bold]Clinical case:[/bold] A 61 year-old woman admitted for a fibular fracture was found to have clinical features of Cushing[apos]s syndrome. She had elevated urinary free cortisol (880 nmol/d, N:[lt]330 nmol/d) with a fasting plasma cortisol of 223 nmol/L and a suppressed plasma ACTH level (0.8 pmol/L, N:2.0-11.0 pmol/L). Adrenal CT and MRI showed bilateral adrenal enlargement and nodules (R: 6.5 x 3.5 cm; L: 8 x 6.9 cm) with heterogeneous features, evocative of mixed AIMAH and myelolipoma. FDG PET scan was not suggestive of malignancy.[br][bold]Investigation:[/bold] In vivo testing for aberrant receptors showed a marked cortisol response to mixed meals (+353%), oral 75 g glucose (+71%), GIP 0.6 mcg/kg/h infusion (+416%) and hLH 300 IU IV (+243%). Endogenous LH was presumably suppressed by narcotic analgesia. Medical therapy with Octreotide to suppress endogenous GIP was unable to control meal induced hypercortisolism; the multi somatostatin receptor ligand Pasireotide was able to reduce post-prandial cortisol, but produced important hyperglycemia. Bilateral adrenalectomy was performed and the left adrenal was predominantly composed of lipomatous tissue with areas of hematopoiesis and strands of adrenocortical nodular hyperplasia; the right adrenal was mainly composed of AIMAH with some foci of myelolipoma. GIP receptor overexpression was confirmed by RT-PCR and immunohistochemistry in adrenocortical hyperplasia tissues mainly in the right adrenal.[br][bold]Conclusion:[/bold] This case is the first report of the coexistence of myelolipoma and AIMAH with GIP/LH dependent Cushing[apos]s syndrome. Their simultaneous occurrence may suggest adrenocortical tissue production under aberrant receptor control of growth factors stimulating adipocyte and hematopoietic proliferation.[br][br]Disclosures: AL: Study Investigator, Novartis Pharmaceuticals; Speaker, Novartis Pharmaceuticals; Advisory Group Member, Ipsen; Speaker, Pfizer, Inc.; Editor, Up To Date; Advisory Group Member, Otsuka, HRA Pharma. Nothing to Disclose: LB, ML, KZ, OP, SO, LMM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 877 65 549 SAT-460 PO45-01 Saturday 488 2012


489 ENDO12L_SAT-461 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Unmasking of Ectopic Cushing Syndrome Following Treatment of Corticotropin Co-Secreting Pheochromocytoma ChenchiReddy Kankara, Steven J Creely, John W Foote, Tabinda Dugal Royal Cornwall Hospital, Truro, UK We report a case of Pheochromocytoma co-secreting ACTH as a rare cause of ectopic Cushing[apos]s syndrome. Our patient is a 49 year lady admitted acutely with sudden onset headache and severe hypertension. CT brain and CSF analysis were normal. Her urinary catecholamines were found to be elevated. Noradrenaline 709 and 972 nmol/mmol of creatinine (ref range 0-48), Adrenaline 215 and 256 nmol/mmol of creatinine (ref range 0-10). CT abdomen revealed 4.2 cm left adrenal mass, MIBG scan confirmed pheochromocytoma. Initial treatment involved alpha and beta blockade with dose titration, reaching up to phenoxybenzamine 70mg three times daily and propranolol LA 160mg daily. Despite this she remained haemodynamically unstable and clinically started to become cushingoid. Investigations for cortisol excess revealed ectopic ACTH secretion. Cortisol following 1mg dexamethasone suppression test was [gt]1750 nmol/L, 24hr urine free cortisol [gt] 1750 nmol/L and ACTH 555 ng/L (ref.range 0-50). Metyrapone was added to her treatment and was transferred to tertiary centre for adrenalectomy. Her pre-op course was complicated by admission to ITU for suspected respiratory failure due to swine flu associated pneumonia and a small pulmonary embolus. After recovery, she successfully underwent laparoscopic left adrenalectomy. Subsequent biochemical testing and imaging confirmed complete resolution of catecholamine and cortisol excess. Screening for MEN was negative.[br]This case is unusual in its acute presentation with highly metabolically active phaeochromocytoma which has probably masked the symptoms and signs of cortisol excess until active blockade of catecholamine action. This case highlights the need to consider the possibility of other hormone dependent syndromes due to various peptide hormones secreted by pheochromocytoma tumor.[br][br]Nothing to Disclose: CK, SJC, JWF, TD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2190 65 550 SAT-461 PO45-01 Saturday 489 2012


490 ENDO12L_SAT-462 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Cushing Syndrome Induced by Ocular Dexamethasone Joanna Mitri, Ronald Lechan Tufts Medical Center, Boston, MA [bold]Background:[/bold] Cushing[apos]s syndrome is a well-known side effect of excess corticosteroids, including oral, topical, intranasal and inhaled. However, Cushing[apos]s syndrome due to topical ocular steroids is extremely rare.[br][bold]Objective:[/bold] To report an unusual cause of Cushing[apos]s syndrome.[br][bold]M[/bold][bold]ethod:[/bold] We present the clinical course of an adult patient presenting with Cushing[apos]s syndrome.[br][bold]Case Report:[/bold] A forty-two year old woman with history of Pott[apos]s disease and long standing history of glaucoma was referred to the endocrine clinic for evaluation of a low AM cortisol, found during the evaluation of extreme fatigue. The patient reported taking dexamethasone eye drops bilaterally 6 times per day for cataracts. Clinically, the patient appeared cushinoid including centripital obesity with a BMI of 45 kg/m[sup]2[/sup], proximal myopathy, ecchymosis and wide violatious, abdominal striae. Biochemical evaluation revealed a serum cortisol of [lt]0.2 mg/dl concomitantly with an ACTH of 5 pg/ml and dexamethasone level 66 ng/dl (normal less than 20ng/dl). Hemoglobin A1C was 5.7%, consistent with pre-diabetes. After gradually decreasing the dexamethasone eyedrops to once per day, her cushinoid features gradually faded and adrenal function improved with an AM cortisol of 9.3 mg/dl.[br][bold]Discussion:[/bold] Exogenous glucocorticoid use is often apparent from the medical history. However, on rare occasions, the clinician may be faced with a case of surreptitious use of glucorticoids or unusual methods of administration. In this patient, a low basal cortisol level and a low ACTH confirmed the presence of a depressed hypothalamic-pituitary-adrenal axis. However, detectable levels of dexamethasone in the blood in the context of cushinoid features strongly suggested that dexamethasone eye drops were responsible for the adrenal axis suppression. There are only 3 cases described in the literature of Cushing[apos]s syndrome related to glucocorticoid ocular drops (1-3). In the latter report, co-administration of ritonavir was responsible for the accumulation of excessive systemic levels of topical ocular steroids.[br][bold]Conclusion:[/bold] Prolonged, unsupervised use of glucocorticoid occular drops may result in Cushing[apos]s syndrome and its numerous associated complications. Physicians should be aware of this possibility and must educate patients about the potentially serious adverse effects of such use.[br][br]1. Afandi B, Toumeh MS,Saadi HF. Cushing[apos]s syndrome caused by unsupervised use of ocular glucocorticoids. Endocr Pract 2003; 9: 526-529. 2. Chiang MY, Sarkar M, Koppens JM, et al. Exogenous Cushing[apos]s syndrome and topical ocular steroids. Eye (Lond) 2006; 20: 725-727. 3. Molloy A, Matheson NJ, Meyer PA, et al. Cushing[apos]s syndrome and adrenal axis suppression in a patient treated with ritonavir and corticosteroid eye drops. AIDS 2011; 25: 1337-1339.[br][br]Nothing to Disclose: JM, RL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 615 65 551 SAT-462 PO45-01 Saturday 490 2012


491 ENDO12L_SAT-463 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Iatrogenic Cushing Syndrome with Adrenal Suppression in HIV-Infected Patient Receiving Inhaled Fluticasone and Ritonavir Omar Zmeili, Wael Taha, Abdul Abou-Samra Wayne State University, Detroit, MI [bold]Introduction: [/bold]Ritonavir, a protease inhibitor (PI), is commonly used in the treatment of HIV infection. It is a potent inhibitor of cytochrome P450-3A4 (CYP3A4) activity and consequently decreases the clearance of synthetic glucocorticoid. The coadministration of ritonavir and inhaled fluticasone, a substrate of hepatic CYP3A4, results in corticosteroid accumulation. This may cause Cushing[apos]s syndrome with adrenal suppression.[br][bold]Clinical Case: [/bold]A52-year-old HIV-infected woman receiving antiretroviral therapy including ritonavir for 3 years presented for evaluation of weight gain and facial swelling. The patient had chronic obstructive pulmonary disease and was started on inhaled fluticasone one year prior to presentation. She developed weight gain (22.0 Kg) in 5 months, facial swelling, and lower extremity edema. On clinical examination the patient was markedly cushingoid with a body mass index (BMI) of 43.5 kg/m[sup]2[/sup], moon face, buffalo hump, truncal obesity, and purple striae over both arms. Laboratory evaluation revealed low morning cortisol [lt] 0.5 mcg/dL (normal range, 4.0-22.0) and low ACTH [lt]0.5 pg/mL (normal range, 6.0-50.0). The 24-h urinary free cortisol was very low as well. Iatrogenic Cushing[apos]s syndrome with adrenal suppression was diagnosed. Fluticasone was discontinued and hydrocortisone replacement was started. After 3 months, there was significant improvement of the moon face and the striae over both arms. At 3 months after fluticasone cessation, the early morning cortisol was 12.2 mcg/dL and ACTH was 41.0 pg/mL. Recovery of endogenous adrenal function was confirmed by cosyntropin stimulation test with a cortisol level rise from 10.0 to 18.3 mcg/dL. She continued to do well after cessation of hydrocortisone.[br][bold]Conclusion: [/bold]Iatrogenic Cushing[apos]s syndrome should be considered in HIV-infected individuals receiving ritonavir and inhaled steroids. Careful evaluation and management are needed to prevent adrenal crisis.[br][br]Nothing to Disclose: OZ, WT, AA-S 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 903 65 552 SAT-463 PO45-01 Saturday 491 2012


492 ENDO12L_SAT-464 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Ectopic Cushing Syndrome: A Difficult Diagnosis Candice Rose, Leigh M Eck University of Kansas, Kansas City, KS [bold]Introduction:[/bold] Ten to 50 percent of patients with ectopic Cushing[apos]s syndrome (CS) have undergone inappropriate transphenoidal surgery. Higher rates are found among those patients with occult sources. This may be at least partially related to increased rates of pituitary incidentalomas attributed to improvements in imaging. It is often difficult to localize a small and indolent tumor. We present a case of misdiagnosed ectopic CS related to a bronchial carcinoid tumor.[br][bold]Clinical case:[/bold] A 28-year old woman presented with rapidly increasing central obesity and abdominal striae. A diagnostic work up revealed ACTH dependent CS that was thought to be due to Cushing[apos]s disease (CD). Given this presumed diagnosis, the patient underwent pituitary resection on two occasions. This rendered her with hypopituitarism, but did not cure her CD. She subsequently underwent bilateral adrenalectomy. ACTH remained elevated but stable following this intervention. Eleven years later she was found to have prominent hyperpigmentation and a rapidly increasing ACTH level with a peak value of 1032 pg/mL (10-60 pg/mL). Due to this rising ACTH level and historical diagnosis of CD, there was concern for Nelson[apos]s syndrome. MRI of the pituitary was performed without any evidence of mass in the pituitary fossa. On further retrospective analysis, a review of the surgical pathology from the decade[apos]s prior hypophysectomies showed no evidence of ACTH staining cells. At this point, the index of concern for a previously undiagnosed ectopic ACTH secreting tumor was high. Imaging was pursued; a CT chest revealed a pulmonary nodule measuring 1.6 x 1.7 cm and an octreotide scan confirmed this nodule to be somatostatin avid. Surgical resection of the nodule was undertaken with pathology consistent with an atypical carcinoid tumor.[br][bold]Clinical lessons:[/bold] The diagnosis and differential diagnosis of CS is one of the most challenging algorithms in endocrinology. ACTH dependent CS is CD in the majority of cases; but, even though rare, an ectopic source of ACTH must be considered in all cases. Multiple modalities including dynamic hormonal testing [CRH stimulation and high-dose dexamethasone suppression testing], imaging [pituitary MRI with gadolinium], and invasive testing [bilateral inferior petrosal sinus sampling] should be undertaken with results analyzed together when determining an etiology of ACTH dependent CS.[br][br]Nothing to Disclose: CR, LME 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 288 65 553 SAT-464 PO45-01 Saturday 492 2012


493 ENDO12L_SAT-465 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Clinical Recovery in Patients with Severe Deconditioning from Cushing Syndrome Susmeeta T Sharma, Nicola M Neary, Lynnette K Nieman Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD [bold]Background: [/bold]Proximal muscle weakness in Cushing[apos]s syndrome (CS) can lead to decreased mobility and functional capacity. Clinical recovery during remission is a slow and sometimes incomplete process. We describe the clinical course of five patients with ACTH-dependent CS with extreme muscle weakness and inability to perform activities of daily living (ADLs).[br][bold]Case Series:[/bold] Four patients (3 women, 1 man) had ectopic ACTH syndrome (EAS), one man had Cushing[apos]s disease. Age at presentation was 38-72 years and duration of symptoms 2.5-30 months. All patients were previously fully independent in ADLs. Progressive muscle weakness, vertebral fractures and depression associated with CS led to decreased mobility. On admission, three patients had 0-2/5 strength in lower extremities (LE) requiring maximal assistance in all ADLs. Two patients had 4-5/5 LE strength. One was limited in ADLs due to back pain; LE strength in the other patient progressively deteriorated (0-2/5) during admission. Serum and urine cortisol levels were elevated and creatine phosphokinase, creatinine, vitamin D, total protein and albumin were low or low-normal in all. Imaging studies showed diffuse fatty infiltration of muscles (n=3) and vertebral fractures (n=4). All patients were wheelchair or bed-bound after prolonged hospitalization (25-155 days) including intensive care (1-153 days). Physical therapy was initiated early and continued throughout hospitalization except during medical instability. Spinal brace was used to stabilize vertebral fractures. Hypercortisolism resolved after surgical resection in three cases [pituitary adenoma, pulmonary and thymic neuroendocrine tumors (NET)]. One patient with metastatic NET underwent bilateral adrenalectomy (BilAdx). Despite control of hypercortisolemia with etomidate and later BilAdx, one EAS patient died from intestinal perforation after a 5 month admission. Autopsy revealed an ACTH-staining pancreatic NET with gastrointestinal metastases. The other patients continued physical therapy after discharge, in an acute care rehabilitation facility or as an outpatient. After 7-12 months of continued physical therapy, they were able to walk with or without an assistive device and were independent in other ADLs.[br][bold]Conclusion: [/bold]Cushing[apos]s syndrome can be associated with severe prolonged muscle weakness, which may improve after resolution of hypercortisolism. We recommend early initiation and continuation of physical rehabilitation to aid clinical recovery.[br][br]Sources of Research Support: Research was funded in part by the intramural program of the Eunice Kennedy Shriver National Institute of Child Health and Human Development.[br][br]Nothing to Disclose: STS, NMN, LKN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 889 65 554 SAT-465 PO45-01 Saturday 493 2012


494 ENDO12L_SAT-466 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Cushing Disease and Opportunistic Infections Paula PB Silva, Lenira C Stella, Fadlo F Filho, Gabriella D Arcari, Isabela TI Carvalho, Julia F Magalhaes Beneficencia Portuguesa, Sao Paulo, Brazil Hypercortisolemic patients, both those with endogenous Cushing[apos]s syndrome, and more common those receiving exogenous glucocorticoid therapy, are at risk of opportunistic infections, such as cryptococcosis, asperfillosis and nocardiosis. High levels of cortisol impacts the immune system, reducing, mainly cellular immunity, by the hormonal action on neutrophiles, macrophages and lymphocytes Th1, with significant increase on the susceptibility to opportunistic infections, especially the fungal ones. These affects are manifested clinically by a decreased cutaneous expression of delayed hypersensitivity, masking of the clinical signs of inflammation, and predisposition to infection. Normally, it impacts the system in a dose-dependent way, with higher chances of serious and opportunistic infections in cases with higher values of plasmatic cortisol. A 38-years-old woman, came to the clinic for Hipothyroidism and Dyslipidemia, she also related headache, rapid weight gain, hirsutim and proximal muscle weakness for 5 years, on progress. After being examined, she had cushingoid facies and truncal obesity. Her blood pressure measured 110 x 70 mmHg, the pulse was regular with a frequency of 76 BPM, and her BMI was 26,8 kg/m2. Laboratory evaluation demonstrated glicemia 101 mg/dL, HbA1c 5,9%, total cholesterol 244 mg/dl, HDL 73 mg/dl, LDL 148 mg/dl, PTH 99,2 pg/ml, Ca 8,5 mg/dl, TSH 1,66 mIU/Ll, T4l0,87 ng/dL. The disease was confirmed by increased excretion of urinary free cortisol (262,4[micro]g/24h) and a failure of suppression of plasma cortisol by a low dose of dexamethasone(16[micro]g/dL l). Plasma ACTH was 5,0 pg/mlL. A CT scan of the abdomen disclosed a nodular lesion of 2,9 cm in diameter in the left adrenal, with density of 18 HU. The chest X- ray showed an infitrate with two nodular densities in the bottom part of the right lung. She went to the surgery of left adrenelectomy and the ressection of the part of the right lung, the material revealed granulomatous inflamation containning cyptococci. The patient was treated with antifungal therapy. In early reports on patients with hypercotisolism it was already recognised that opportunistic infections were an important cause of morbilidy and mortality in those patients. Other cases of Cushing syndrome and pulmonary cryptococcosis are related in literature. This case report calls attention to need for recognition this patients which presents weight gain instead caquexia like tipical fungic infections.[br][br]Bakker et al., J Endocrinol Invest 1998;21(5):329:33. Graham BS et al., Ann Intern Med 1984; 101(3)334:8. Kramer et al, Arch Inter Med 1983; 143(11): 2179-80.[br][br]Nothing to Disclose: PPBS, LCS, FFF, GDA, ITIC, JFM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1989 65 555 SAT-466 PO45-01 Saturday 494 2012


495 ENDO12L_SAT-467 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Latent Autoimmune Hepatic Dysfunction Unmasked by Treatment of Endogenous Hypercortisolemia Diala El-Maouche, Susmeeta Sharma, Lynnette Nieman National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD Background: Endogenous hypercortisolemia of Cushing[apos]s syndrome (CS) can be associated with improvement of autoimmune diseases with a risk of exacerbation upon remission.[br]Clinical Case:[br]A 26 year old woman diagnosed with CS in 2007, and subsequently developed transaminitis during ketoconazole (KTZ) treatment. Later that year she had remission after left lower lobe wedge resection of an ACTH-secreting neuroendocrine tumor with lymph node metastases. She remained in remission until 2010. Steroidogenesis inhibitors were given when the source(s) of ACTH could not be found and patient declined bilateral adrenalectomy. She developed severe neutropenia on combination therapy with metyrapone and KTZ, leading to their discontinuation. Use of mifepristone, a glucocorticoid antagonist, was associated with increased ACTH and cortisol levels and worsening peripheral edema, weight gain, and hypertension. Remission (cortisol [lt]1-2.5 mcg/dl, normal 5-25) was achieved in February 2011 on a block and replace regimen with octreotide, KTZ (600 mg/day) and dexamethasone 0.25 mg/day. In May 2011, laboratory tests first showed elevated transaminases with ALT 160 (normal 0-40 IU/L) and AST 117 (normal 0-40 IU/L). Ketoconazole was stopped due to concern for drug-induced hepatitis. Octreotide was discontinued three weeks later after persistent rise in transaminases (ALT 427, AST 302 IU/L) off KTZ. Further evaluation showed a normal liver ultrasound, normal alkaline phosphatase and bilirubin levels, negative hepatitis serology, positive anti-mitochondrial antibody (AMA) and negative anti-smooth-muscle antibody. She remained asymptomatic. Autoimmune hepatitis and early primary biliary cirrhosis were considered as possible causes given positive AMA and increasing transaminases weeks after discontinuation of KTZ. Liver biopsy showed periductal inflammation and lobular hepatitis. Prednisone 30 mg/day was initiated in late June 2011 when transaminases continued to rise (ALT 839, AST 579 IU/L). Transaminitis resolved and prednisone was gradually tapered off in September 2011. Liver function tests remained normal off prednisone in the setting of recurrent endogenous hypercortisolemia (UFC=90 mcg/day, normal 3.5-45).[br]Conclusion: Treatment of endogenous hypercortisolemia in CS can be associated with unmasking of latent autoimmune disease. Autoimmune hepatitis should be considered in the differential diagnosis of hepatic dysfunction in a eucortisolemic CS patient on KTZ.[br][br]Sources of Research Support: This work was funded in part by the intramural program of NICHD, NIH (The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health).[br][br]Nothing to Disclose: DE-M, SS, LN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 666 65 556 SAT-467 PO45-01 Saturday 495 2012


496 ENDO12L_SAT-468 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Small-Cell Carcinoma of the Endometrium Presenting as Cushing Syndrome Haruhiro Sato, Masafumi Fukagawa, Yoshiyuki Robert Osamura Tokai University School of Medicine, Shimokasuya, Isehara, Japan; Tokai University School of Medicine, Shimokasuya, Isehara, Japan Small-cell carcinoma (SCC) of neuroendocrine type is an uncommon tumor of the endometrium. No previous report has documented Cushing[apos]s syndrome due to ectopic ACTH production by SCC of the endometrium. We describe a 56-year-old Japanese woman with SCC of the endometrium and multiple lung metastases presenting as Cushing[apos]s syndrome. The patient was referred to our hospital because of general fatigue with facial and leg edema, and multiple nodular lesions in the bilateral lungs on chest X-ray examination. A physical examination revealed that the patient had moon face, buffalo hump, and truncal obesity. Laboratory tests performed at 0800 h showed a plasma ACTH level of 234 pg/ml (normal range, 7.4-55.7), a serum cortisol level of 54.0 [micro]g/dl (normal range, 4.0-18.3), and a serum dehydroepiandrosterone sulfate (DHEAS) level of 2730 ng/ml (normal range, 80-1880). The urinary free cortisol level was 1940 [micro]g/day (normal range, 11.2-80.3). Circadian rhythm of ACTH and cortisol had been lost. After an overnight low-dose (1 mg) dexamethasone suppression test, her serum cortisol level was 69.2 [micro]g/dl (normal response, [lt]5). After an overnight high-dose (8 mg) dexamethasone suppression test, the serum cortisol level was 83.0 [micro]g/dl (less than 50% of baseline in Cushing[apos]s disease). Endocrinological examinations confirmed ACTH-dependent Cushing[apos]s syndrome. Thoracic computed tomography imaging showed multiple nodular lesions in the bilateral lungs. Abdominal magnetic resonance imaging suggested a malignant tumor of the uterus. The patient received a lung tumor biopsy and surgical hysterectomy. The endometrial carcinoma was histologically a SCC admixed with endometrioid adenocarcinoma. The SCC of the endometrium showed immunoreactivity for pro-opiomelanocortin, ACTH, and vimentin, but not for thyroid transcription factor-1 (TTF-1). The lung biopsy specimen had the same features. The tumor in the endometrium was macroscopically solitary, whereas CT imaging revealed multiple tumors in the lung, suggesting that the latter were metastases. TTF-1 is widely used in the diagnosis of thyroid and lung carcinomas. In the present case, both the SCC and endometrioid adenocarcinoma components showed no TTF-1 immunoreactivity. These findings indicated that the SCC originated from the endometrium, and the ectopic ACTH-producing tumor caused Cushing[apos]s syndrome. This study provides the evidence that SCC of endometrial origin was an ectopic ACTH-producing tumor causing Cushing[apos]s syndrome.[br][br]Nothing to Disclose: HS, MF, YRO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1571 65 557 SAT-468 PO45-01 Saturday 496 2012


497 ENDO12L_SAT-469 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Ectopic ACTH Syndrome Due to MEN-I Associated Metastatic Neuroendocrine Pancreatic Gastrinoma in a 22 Year Old Female without Hyperparathyroidism: Diagnostic and Therapeutic Challenges Mansoor Tanwir, Vasudev G Magaji, Korytkowski Mary, Challinor M Sue University of Pittsburgh, Pittsburgh, PA [bold]Introduction: [/bold]Ectopic ACTH production due to malignant tumors is a rare cause of Cushing[apos]s syndrome (CS). Diagnosis and treatment of such patients can be challenging.[br][bold]Case: [/bold]22 year old female with recent onset fatigue, weight gain, polyuria, polydipsia, presented with melena and hypotension. She had cushingoid facies, acne, central obesity, muscle weakness and purple striae. Labs showed: Cortisol 56 [micro]g/dl (5-15), 24 hour UFC [gt]1000 [micro]g/24hr (5-50), Cortisol after 1mg Dexamethasone suppression 54 [micro]g/dl ([lt]1.8), ACTH 211 pg/ml (5-27), Ca 9.1 mg/dl (8.5-10.3), and PTH 25 ng/dl (11-54). EGD showed multiple gastric ulcers and gastrin level was 1599 ng/L ([lt]200) for which PPI and octreotide were started. Her cortisol level did not suppress with 8mg Dexamethasone (baseline 58.8 ng/dl, post supression 132.2 ng/dl), suggesting ectopic ACTH source. CRH stimulation test resulted in 26% and 33% rise in ACTH and Cortisol level respectively, indicating possible pituitary source. CT scans revealed a 5cm hepatic cavernous hemangiona, which on MRI was defined as 6 cm metastatic lesion. Pituitary MRI also revealed a 3mm adenoma. IPSS revealed no Central:Peripheral gradient, confirming ectopic ACTH production. Endoscopic ultrasound (EUS) revealed a 2cm pancreatic lesion not seen on CT or MRI. Biopsy confirmed pancreatic neuroendocrine tumor (PNET) with rare staining for gastrin and ACTH. Ketoconazole was started and discontinued following elevations in transaminase. Etomidate infusion at 0.2mg/kg IBW was used to maintain Cortisol of 20-30 ng/dl until Metyrapone was available. Hospital course was complicated by multiple pulmonary emboli with hemodynamic compromise requiring thrombolysis and anticoagulation, delaying surgical intervention. MEN-1 testing was positive, despite a negative family history and absence of hyperparathyroidism. Currently, she is maintained on Metyrapone with normal UFC levels.[br][bold]Learning points:[/bold][br]When evaluating patients with suspected ectopic ACTH production, independent review of radiologic imaging should be done to avoid being misled by prior reports. EUS is a superior method for identifying PNET (1). Biochemical confirmation with IPSS helps exclude Cushing[apos]s Disease in the setting of a pituitary adenoma. Medical therapy to control hypercortisolism and anticoagulation are necessary to lower morbidity and mortality associated with severe CS (2). Finally, screening for MEN-1 in these patients is recommended, even in the absence of hyperparathyroidism (3).[br][br]1)Anderson MA et al., Am J Gastroenterol. 2000; 92: 2271-7. 2)Isidori AM et al., J Clin Endocrinol Metab. 2006; 91: 371-377. 3)Brandi ML et al., J Clin Endocrinol Metab. 2001; 86: 5658-71.[br][br]Nothing to Disclose: MT, VGM, KM, CMS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1990 65 558 SAT-469 PO45-01 Saturday 497 2012


498 ENDO12L_SAT-470 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Pituitary Macroadenoma Disappearance with Ketoconazole Treatment: An Unusual Case of Cushing Syndrome with Interesting Findings Saira Furqan, Asma Ahmed, Najmul Islam Aga Khan University and Hospital, Karachi, Pakistan Introduction:[br]ACTH dependent Cushing[apos]s is usually associated with diagnostic dilemma especially when it occurs concomitantly with pituitary adenoma. We report a case of ACTH dependent hypercortisolism with concomitant pituitary macro adenoma and chest lesions. [br]Case Summary:[br]This is a case history of 38 year old male patient with complains of significant weight loss and proximal myopathy. Investigations revealed significant hypokalemia and biochemical hypercortisolism insuppressible with both low and high doses of dexamethasone. Serum ACTH level was found to be 60 (7-63). Diagnosis of ACTH dependent Cushing[apos]s syndrome was made. MRI Brain showed pituitary macro adenoma (2[times]1.9cm). IPSS was done to confirm central/pituitary source, but the study failed to show any significant gradient. Subsequently CT Scan Chest and Abdomen was also done which revealed necrotic mass lesion seen in right upper lobe (4x2.9cm). Multiple tiny nodules in right lung, largest one measuring 7 mm, was also noted. As the source of ACTH was not evident the patient was started on ketoconazole, the dose was gradually increased to 1200mg/day. Spironolactone with potassium replacement was also started. For the chest lesion broad spectrum intravenous antibiotics was given for 2 weeks. After 1 month CT scan of chest and abdomen were repeated which showed significant reduction in the size of soft tissue density mass noted in right upper lung (11x10 mm, previously 43 x 34mm) with complete resolution of right sided lesions but persistent mediastinal lymph nodes. Mediastnal LN biopsy via VATS was done which showed benign reactive changes One month after mediastinal lymph node biopsy, repeat chest X-ray was completely normal with no abnormal finding. Due to normal biopsy, the main source was thought to be of pituitary macro adenoma. Repeat MRI of pituitary was done which showed complete resolution of adenoma and the dose of ketoconazole has been gradually reduced.[br]Conclusion:[br]Ketoconazole not only resulted in biochemical suppression of Cushing[apos]s but it also caused disappearance of pituitary macro adenoma.[br][br]Nothing to Disclose: SF, AA, NI 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1648 65 559 SAT-470 PO45-01 Saturday 498 2012


499 ENDO12L_SAT-471 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Cushing Syndrome in Pregnancy Karen Hutchinson, Kunchang Song, Stephen Hammes, Loralei Thornburg, Tulin Ozcan, Jacob Moalem, Runhua Hou University of Rochester, Rochester, NY; University of Rochester, Rochester, NY; University of Rochester, Rochester, NY; University of Rochester, Rochester, NY [bold]Introduction[/bold][br]Cushing[apos]s syndrome during pregnancy is uncommon. During normal pregnancy the hypothalamic-pituitary-adrenal axis is affected, resulting in elevated total cortisol, CBG and ACTH. Also, many of the signs and symptoms of cortisol excess can overlap with normal features of pregnancy making the diagnosis difficult.[br][bold]Clinical Case[/bold][br]A 29-year-old woman with no previous significant medical history presented to endocrine clinic at 34 weeks gestation with hypertension, gestational diabetes and 60 pound weight gain during pregnancy. She had diffuse purple striae of her arms, breasts, abdomen and upper legs in addition to emotional lability, facial plethora, supraclavicular fat pads and proximal muscle weakness. She also complained of headaches and had proteinuria. The patient was admitted to the hospital for evaluation for preeclampsia and Cushing[apos]s syndrome. While in the hospital, her 8 am cortisol level was elevated at 37.4 ug/dL (6.2-19.4 ug/dL) with a corresponding suppressed ACTH of [lt]5 pg/dL (0-46 pg/dL). On two consecutive nights, midnight salivary cortisol levels were high at 605 [amp] 1020 ng/dl ([lt]100 ng/dl). Her 24-hour urinary cortisol was high on 2 separate occasions, with amounts of 549 [amp] 468 ug/24h ([lt]45 ug/24h). Based on her gestation age and preeclampsia, the patient underwent induction of labor at 36 weeks and delivered a healthy female infant. Following delivery, her serum cortisol remained high (36.7 [amp] 36.4 ug/dL at 9 am [amp] 11 pm respectively). An abdominal CT scan, showed a 3.1 x 3.2 cm left-sided adrenal mass. The patient underwent a laparoscopic left adrenalectomy, with pathology revealing a 3.2 x 3 x 3 cm round, bright yellow tumor. Cells stained negative for LH, but weakly positive for hCG, which suggest hCG may have stimulated Cushing[apos]s syndrome to progress. She was discharged home on hydrocortisone replacement.[br][bold]Conclusions[/bold][br]Cushing[apos]s syndrome during pregnancy is difficult to diagnose due to normal pregnancy physiologic changes with elevations in cortisol, CBG [amp] ACTH; however, diurnal variation of cortisol should be maintained. In this case, the diagnosis of Cushing[apos]s syndrome was made by the combination of her progressive symptoms and the laboratory evaluation that revealed the loss of diurnal cortisol secretion and markedly elevated urinary cortisol levels. Clinicians should maintain a high index of suspicion for new onset Cushing[apos]s syndrome during pregnancy in any patient with pronounced symptoms suggestive of cortisol excess.[br][br]Nothing to Disclose: KH, KS, SH, LT, TO, JM, RH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 431 65 560 SAT-471 PO45-01 Saturday 499 2012


500 ENDO12L_SAT-472 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Case Report of Cushing Syndrome in Pregnancy: An Unanswered Conundrum Reem Kheetan, Vishnu Garla, Tipu Saleem Marshall University, Huntington, WV Objective: To report a rare case of Cushing syndrome in pregnancy and to elucidate the diagnostic and management dilemmas associated with it.[br]Methods: We present Patient[apos]s clinical history, physical findings, diagnostic work up and treatment along with the literature review.[br]Results: A 31 year old 16 weeks pregnant woman with history of gestational diabetes and hypertension was admitted with symptoms of headaches, blurred vision along with excessive weight gain and hirstuism. Physical examination revealed facial plethora, prominent supraclavicular fat pads, wide purple abdominal striae and multiple bruises in the lower extremities. Review of her medical records revealed a left sided adrenal mass of 2.7 cm found 6 months ago with inconclusive and incomplete endocrine work up. Further laboratory data revealed serum cortisol 24.5 mcg/dl (3-17),24 hour urine free cortisol 1845 mcg/24hours (0-50) and 24 hour urinary protein 585 mg (0-75), ACTH [lt]1.1 pg/ml (7-63). She has normal levels of renin, aldosterone, DHEA and urinary metanephrines. Serum cortisol did not suppress (24 mcg/dl) after overnight 1mg of dexamethasone. MRI revealed that left adrenal mass has enlarged and now measures 3.2 cm with signal dropout on out of phase imaging suggestive of a lipid laden adrenal adenoma. She was diagnosed with cushing syndrome due to left adrenal adenoma and left adrenalectomy was done. Histopathology specimen confirmed the diagnosis without any evidence of malignancy. Patient was started on physiologic dose of steroids. Her requirements for antihypertensive medications and insulin drastically reduced and proteinuria improved after the surgery. Plasma ACTH level rose up to normal after 4 months. Patient received stress dose steroids during an uncomplicated delivery of a healthy male baby after 37 weeks of gestation. Steroids were slowly tapered off after the delivery.[br]Conclusion: Pathologic Cushing syndrome in pregnancy is very rare and poses a diagnostic dilemma to the physician due to the physiologic hyper secretion of cortisol during normal pregnancy.[br][br]1) De P, Evans LM, Scanlon MF, Davies JS. [quot]Osler[apos]s phenomenon[quot]: misdiagnosing Cushing[apos]s syndrome. Postgrad Med J. 2003 Oct;79(936):594-6. 2)Steffensen C, BakAM, Rubeck KZ, Jorgensen JO. Epidemiology of Cushing[apos]s syndrome.Neuroendocrinology. 2010;92Suppl 1:1-5. Epub 2010 Sep 10. 3)Vilar L, FreitasMda C, Lima LH, Lyra R, Kater CE. Cushing[apos]s syndrome in pregnancy: an overview. Arq Bras Endocrinol Metabol. 2007 Nov;51(8):1293-302. 4)Lindsay JR, Nieman LK. The hypothalamic-pituitary-adrenal axis in pregnancy:Challenges in disease detection and treatment. Endocr Rev. 2005 Oct;26(6):775-99. 5)Fayol L, Masson P, Millet V, Simeoni U. Cushing[apos]s syndrome in pregnancy andneonatal hypertrophic obstructive cardiomyopathy. ActaPaediatr. 2004 Oct;93(10):1400-2. 6)Buescher MA, McClamrock HD, Adashi EY. Cushing syndrome in pregnancy.Obstet Gynecol. 1992 Jan;79(1):130-7. 7) Deborah J, Robert H, John D. Cushing[apos]s syndrome in pregnancy. CMAJ,1989 Nov; Vol 141. 8) Stewart PM. The adrenal cortex. In:Kronenberg HM, Melmed S, Polonsky KS, Larsen PR, eds. Williams Textbook of Endocrinology. 11 ed.Philadelphia, PA: WB Saunders, 2008: 445-503. 9)Aron DC, Schnall AM, Sheeler LR. Cushing[apos]s syndrome and pregnancy. Am J ObstetGynecol. 1990 Jan;162(1):244-52. 10) Blanco C, Maqueda E, Rubio JA, Rodriguez A. Cushing[apos]s syndrome during. pregnancy secondary to adrenal adenoma: metyrapone treatment and laparoscopic. adrenalectomy. J Endocrinol Invest. 2006 Feb;29(2):164-7.[br][br]Nothing to Disclose: RK, VG, TS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 413 65 561 SAT-472 PO45-01 Saturday 500 2012


501 ENDO12L_SAT-473 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Cushing Syndrome in Pregnancy Secondary to an Adrenal Adenoma Vivian Cenize Guardia, Ane Caroline The Bonifacio Freire, Nilza Scalissi, Jose Viana Lima Junior, Rachel Emerick Mendes, Leticia Iervolino, Lydia Sedda Souza Irmandade Santa Casa de Misericordia de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Introduction: Pregnancy is uncommon in women with Cushing[apos]s syndrome (CS), with fewer cases reported in literature, as both excessive cortisol and androgen levels suppress gonadotropin secretion and induce ovulatory disturbances. Although it is a rare condition, it increases rates of spontaneous abortion, perinatal death, premature birth and intrauterine growth retardation. Maternal morbidity includes hypertension, preeclampsia, diabetes and opportunistic infections.(1) The clinical diagnosis of CS during pregnancy may be missed because of the overlapping features that occur in normal pregnancy. Also normal physiologic changes in pregnancy, including increases in serum/urinary cortisol and plasma ACTH levels, complicate the screening process. (2)[br]Clinical case: A 25-yr-old woman was admitted in our hospital at 17[sup]th[/sup] gestacional week with a 4-month history of hypertension, weight gain, hirsutism and widespread acne. Physical examination also revealed moon face, facial plethora, proximal muscle weakness, central obesity and few violaceous striae. Serum cortisol at 8 am after 1mg of dexamethasone overnight was not suppressed (25.9 ug/dl), urinary cortisol was within the normal range (32.7 ug/24hs), total serum testosterone was elevated (319 ng/dl), as well as serum androstenedione (more than 10.0 ng/ml), and adrenocorticotropic hormone level was 10.7 pg/ml. On admission, she was also diagnosed with diabetes. After 6 days of hospitalization, the patient had an acute hypertensive pulmonary edema followed by spontaneous abortion. Thereafter, an abdominal CT was performed and detected a mass of 3 cm in the right adrenal gland. After some days, the patient underwent a laparoscopy right adrenalectomy and pathologic examination surprisingly revealed a benign adrenocortical adenoma. Corticoid replacement therapy was gradually reduced during the following month in the post operatory follow-up. Subsequent biochemical tests suggested disease remission and, after 8 months, the patient remains with mild hypertension, but has had significant improvement in the Cushing phenotype and no longer needs hypoglycemic treatment.[br]Conclusion: An increased suspicion for CS is required for the diagnosis of this rare disorder in pregnancy. Therefore, it is important to consider this clinical condition when a pregnant woman develops hypertension and/or hyperglycemia. The early diagnosis would likely facilitate early treatment and result in improved outcomes for both mother and fetus.[br][br](1)Kim HG et al.,J Korean Med Sci 2003;18:444. (2)Lindsay JR, Nieman LK., Endocrine Reviews 2005; 26:775.[br][br]Nothing to Disclose: VCG, ACTBF, NS, JVLJ, REM, LI, LSS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2047 65 562 SAT-473 PO45-01 Saturday 501 2012


502 ENDO12L_SAT-474 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Nocardia Associated with Series of Patients with Ectopic Cushings Azra Rizwan, Aqiba Sarfaraz, Abdul Jabbar, Jaweed Akhter, Najmul Islam Aga Khan University Hospital, Karachi, Pakistan Introduction[br]Cushing[apos]s syndrome results from prolonged exposure to excess glucocorticoids. Ectopic Cushing[apos]s is endogenous ACTH dependant form of Cushing[apos]s associated with markedly raised ACTH, and, subsequently, cortisol levels. Such levels lead to an impaired immune response, setting the stage for the occurrence of bacterial and fungal opportunistic infections. Nocardia is a gram positive bacterial infection caused by aerobic actinomycetes in the genus Nocardia. We report a series of patients diagnosed with ectopic Cushing[apos]s, with growth of Nocardia asteroides from pulmonary secretions. In one of these cases, the manifestations of Cushing[apos]s disappeared with treatment for Nocardia.[br]Clinical Case[br]Two middle aged men presented to the Endocrine clinic in the same year: the first with history of exertional shortness of breath, and weight loss for 1 year, the other with facial swelling, disturbed sleep and lethargy for a month. The third case was a young male who presented with progressive weakness [amp] weight loss for some time. All three had uncontrolled hypertension, high blood sugars [amp] were hypokalemic (K: [bold]2.52, 2.9, 1.5[/bold]; [normal range 3.6-4.0]; 24 hour urine cortisol was elevated at [bold]2000, 27,216 and 9088[/bold] (32-243); ACTH [bold]68.5, 159, 255[/bold][0-48), respectively. Their MRI pituitary was normal, inferior petrosal sinus sampling revealed no central peripheral gradient. CT chest of these subjects demonstrated cavitatory lung lesions; bronchial washings recovered heavy growth of [bold]nocardia. [/bold]CT guided biopsy revealed no malignancy. Antihypertensives, insulin, potassium replacement, ketoconazole [amp] trimethoprim- sulphamethoxole (TS) were initiated. The patients[apos] symptomatology improved [amp] cavitatory lesions resolved with treatment. The primary source for the ectopic cushings remained unknown. The first case later followed a progressively downhill course leading to death. The second case required bilateral adrenalectomy. In the third case, we were able to completely taper off ketoconazole, potassium, insulin [amp] antihypertensives, after starting TS.[br]Conclusion[br]Opportunistic infections are well known to be associated with Cushing[apos]s syndrome, as higher levels of glucocorticoid secretion are found in patients with ectopically produced ACTH. Nocardia of lungs is important differential to consider. This series includes the first case reported in which signs and symptoms of cushings subsided after treatment of Nocardia.[br][br]Nothing to Disclose: AR, AS, AJ, JA, NI 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 662 65 563 SAT-474 PO45-01 Saturday 502 2012


503 ENDO12L_SAT-475 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Explosive Recurrence of Adrenal Carcinoma after Apparent Surgical Cure Ashutosh Pareek, Rachel Bier, Barnett Zumoff Beth Israel Medical Center, New York, NY [bold]Objective[/bold]: To report a case of rapidly recurring metastatic adrenal carcinoma after surgical [uml]cure.[uml][br][bold]Case[/bold]: A 64-year-old female with history of hypothyroidism, surgically removed parathyroid adenoma, diabetes mellitus (DM), and hypertension (HTN) presented with a three-week history of confusion, fatigue, lower extremity edema, parasthesias of the arms and legs, and diffuse ecchymosis. Physical examination was significant for lower extremity muscle weakness and facial puffiness; no signs of virilization. Laboratory findings included potassium 3.2mmoL/L (3.5-5.1), glucose 161mg/dL (74-106), TSH 0.87mU/L (0.55-4.78) and a random cortisol of 29ug/dL (4-22). Persistent hypokalemia and recent onset of DM and hypertension prompted an abdominal CT scan that showed a 9.5 X 10.8 X 2.5 cm left adrenal mass. Patient underwent left adrenalectomy. Pathology confirmed a T3NOMO adrenal carcinoma with clear tumor margins. Post-op complications included pulmonary embolism requiring anticoagulation and fluid in the surgical bed requiring drainage and antibiotics. Patient[apos]s clinical signs and symptoms showed almost complete resolution within two weeks after surgery, at which time plasma cortisol was 10ug/dL.[br]Three weeks later, cortisol was elevated at 33ug/dL, and a week after that it was 24ug/dL and did not suppress with dexamethasone. Plasma ACTH level was 5pg/ml (5-27) at that time. Due to a strong suspicion of tumor recurrence a PET/CT scan was performed; it revealed pulmonary, hepatic, and sacral metastases only nine weeks after adrenalectomy. There was also a recurrence of all her pre-op signs and symptoms. The patient was subsequently treated with maximal doses of mitotane, fluconazole, and chemotherapy, but there was no improvement and she died soon after.[br][bold]Discussion[/bold]: Adrenal carcinoma is a rare malignancy with a poor prognosis. Five-year disease-free survival after an ostensibly complete resection of a stage I-III tumor is approximately 30%. This patient was biochemically [uml]cured[uml] after adrenalectomy and her Cushingoid clinical picture virtually completely resolved within two weeks. Despite these good results, her tumor advanced explosively and her Cushing[apos]s syndrome promptly recurred. Even with multidrug treatment she deteriorated rapidly and died five months later.[br][bold]Conclusion[/bold]: In patients with adrenal carcinoma, apparent surgical cure does not preclude rapid recurrence and metastasis; therefore, prognosis should be extremely guarded even in best-case scenarios.[br][br]Nothing to Disclose: AP, RB, BZ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 828 65 564 SAT-475 PO45-01 Saturday 503 2012


504 ENDO12L_SAT-476 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) [[sup]18[/sup]F]Fluorodeoxyglucose Positron Emission Tomography-Guided Therapy in Metastatic Adrenocortical Carcinoma: An Illustrative Case Ferhat Deniz, Hubert H Chuang, Kanishka Sircar, Camilo Jimenez, Christopher G Wood, Carlos Rubin De Celis, Mouhammed Amir Habra The University of Texas MD Anderson Cancer Center, Houston, TX; Gulhane Military Medical Academy, Ankara, Turkey; The University of Texas MD Anderson Cancer Center, Houston, TX; The University of Texas MD Anderson Cancer Center, Houston, TX; The University of Texas MD Anderson Cancer Center, Houston, TX; Texas Oncology, PA, Austin, TX [bold]Context:[/bold] [[sup]18[/sup]F] Fluorodeoxyglucose (FDG)-positron emission tomography/computed tomography (PET/CT) scans have been used in staging and monitoring therapy for many cancers, but its role in assessing responses of adrenocortical carcinoma (ACC) to chemotherapy has not been described (1-4).[br][bold]Subject and Methods:[/bold] A 38-year-old man with metastatic ACC for whom neoadjuvant chemotherapy was the initial treatment. Contrast CT and PET/CT scans were obtained before and after chemotherapy. CT response was assessed using Response Evaluation Criteria in Solid Tumors, version 1.1 (RECIST 1.1) and PET response was assessed by PET Response Criteria in Solid Tumors, version 1.0 (PERCIST) (5,6).[br][bold]Results:[/bold] At presentation, the patient had a 14-cm left adrenal mass with para-aortic lymphadenopathy, a tumor thrombus extending into the inferior vena cava, and hepatic metastases. The primary tumor and para-aortic lymph nodes had standardized uptake value-lean (SUL) peaks of 7.1 and 2.7, respectively; the hepatic metastases were more subtle, with representative SUL peak measurements of 2.1 and 1.9. After five cycles of chemotherapy, the patient had a partial response based on RECIST 1.1, whereas he had a complete metabolic response based on PERCIST criteria. Based on these findings, neoadjuvant therapy was halted and he had resection of primary tumor followed by 2 cycles of adjuvant therapy to consolidate response. He remained disease-free 28 months after initial diagnosis.[br][bold]Conclusion:[/bold] FDG-PET/CT scans can be used to assess response to neoadjuvant chemotherapy and yield information useful in planning the surgical intervention. Further studies are needed to clarify the exact role of FDG-PET/CT in the management of ACC.[br][br](1)Boland GW et al.,Radiology 1995; 194:131. (2)Erasmus JJ et al. AJR Am J Roentgenol 1997; 168:1357. (3)Leboulleux S et al. J Clin Endocrinol Metab 2006; 91:920. (4)Antoch G et al. J Clin Oncol 2004; 22:4357. (5)Eisenhauer EA et al. Eur J Cancer 2009; 45:228. (6)Wahl RL et al. J Nucl Med 2009; 50 Suppl 1:122S.[br][br]Nothing to Disclose: FD, HHC, KS, CJ, CGW, CRDC, MAH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 18 65 565 SAT-476 PO45-01 Saturday 504 2012


505 ENDO12L_SAT-477 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Feminizing Adrenal Carcinoma Presenting with Heart Failure and Ventricular Tachycardia Anjana Harnoor, Robert L West, Fiona J Cook Brody School of Medicine at East Carolina University, Greenville, NC [bold]Background[/bold]: Adrenocortical carcinomas (ACC) are rare. Adrenal steroid hormone excess is evident in 40-60% of cases. Feminizing ACC comprise only 1-2%. Estrogen reduces potassium channel currents, causing repolarization to be delayed, which increases susceptibility to arrhythmia. We present a case of feminizing ACC associated with a very high serum estradiol who presented with congestive heart failure and ventricular arrhythmia.[br][bold]Clinical Case[/bold]: A 54 year old male presented with acute heart failure and non-sustained ventricular tachycardia. Echocardiogram revealed dilated cardiomyopathy with ejection fraction 35-40%. EKG showed NSR with a prolonged QTc interval. He had weight loss, decreased libido, erectile dysfunction, and gynecomastia. Physical exam revealed gynecomastia, a hyperdynamic precordium, and a firm mass in the left upper quadrant. Imaging revealed a mass (12 x 11 x 17 cm) felt to arise from the left adrenal gland. Hormonal evaluation revealed a serum estradiol of 3853 pg/ml ([lt]52 males) and mildly elevated adrenal androgens and 24 hour urine free cortisol. He underwent adrenalectomy with subsequent normalization of estradiol level to [lt]30 pg/ml. Surgical pathology was consistent with ACC MacFarlane stage II. On 15 month follow up he had a normal estradiol level, normalization of QTc interval and no further episodes of ventricular arrhythmia. Echocardiogram revealed improvement in cardiac function with an ejection fraction of 60-65%.[br][bold]CONCLUSION: [/bold]Although feminizing adrenal cancers are rare, a middle aged man presenting with new onset gynecomastia should be evaluated with an estradiol level. The effects of very high levels of estrogen on the heart are still poorly understood. Our case highlights the possibility that the uniquely high serum level of estradiol due to adrenal carcinoma contributed to arrhythmia by prolonging QT interval, which normalized with normalization of serum estradiol level after surgery. It is also possible that the very high concentration of estrogen, along with mild elevation of other adrenal steroids, contributed to his dilated cardiomyopathy, which also resolved with adrenal tumor resection. This unique clinical model appears to support previous observations from in vitro and animal studies on the cardiac effects of estrogen and adds to our limited knowledge of this issue in humans.[br][br]Nothing to Disclose: AH, RLW, FJC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 26 65 566 SAT-477 PO45-01 Saturday 505 2012


506 ENDO12L_SAT-478 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Long-Term Survival in a Patient with a Large Adrenal Cortical Carcinoma Producing Cortisol and Androgens Mohammed NMN Ahmed, Rashid NMN Al-Muqbali King Faisal Specialist Hospital [amp] Research Center, Riyadh, Saudi Arabia Background:[br]ACC is a rare endocrine malignancy w/estimated worldwide incidence 0.5- 2/million/yr. It is characterized by a high risk of recurrence; even after radical surgery up to 85% pts. develop recurrence. A decreased OAS (30 %) is reported in most series.Tumors localized to adrenal gland (McFarlane stages 1 [amp]2) have better prognosis than invasive [amp] metastatic tumors (stages 3 [amp] 4).Important factors dictating outcome include, pts.[apos] age, tumor stage, resection status at initial surgery, overexpression of molecular markers (IGF-2, constitutive activation of beta-caten) [amp] adjuvant therapy administered. Whether use of mitotane is beneficial as adjunct Rx remains controversial. Long-term survival of a decade or more is limited to few cases. Little information is available regarding survival status of pts. w/ACC producing both cortisol [amp] andogens.[br]Case Summary:[br]A 21-yr old lady w/amenorrhea, [amp] hirsutism presented in 2000. O/E she had acne, hirsutism(Ferrimen-Gallawey score 3) w/prominent Cushingoid features. Labs: Serum AM/PM cortisol (776/856 nmol/l:nl:-170-356/125-220), Urine free cortisol (UFC) 604 nml/d (nl: [lt]250),Serum ACTH [lt] 10 ng/l (nl:5-60),testosterone 16 nmol/l (nl, female: 0.22-2.9),DHEAS 53.6 umol/l (nl 2.2-15.2).CT abdomen/Chest/Pelvis: 12.5x 10.5 cm lt. adrenal mass, w/out regional or distant metastases. She underwent laparoscopic complete resection. Histology: findings compatible w/ACC 19x11x 9 cm, weighing 643 grams, w/out extracapsular extension. Adjuvant mitotane Rx was started [amp] is continued to date (3 G/D), w/prednisone 5 mg/D. Serum mitotane monitoring has not been available. No other Rx was given.[br]FU Course: For the last 11 yrs. she has been under surveillance using Q 6-12 monthly CT, US abdomen, liver function tests, serum androgens, cortisol [amp] UFC; these [amp] all other labs have remained unremarkable with no evidence of recurrence or metastases, nor any side effects of mitotane Rx. She is fully functional as a teacher [amp] became pregnant x3 during which she withheld mitotane for cumulative 30 mos. She lost one pregnancy but 2 other pregnancies were normal [amp] is raising 2 healthy children.[br]Conclusions.[br]Favorable factors for disease-free survival in our pt. included young age, low-stage tumor, complete resection, lack of metastases, and possibly use of mitotane. Adjunct mitotane may prolong recurrence-free survival in pts. w/radically resected ACC producing cortisol [amp] androgens. Mitotane should be considered in most ACC pts.[br][br]Nothing to Disclose: MNMNA, RNMNA-M 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 329 65 567 SAT-478 PO45-01 Saturday 506 2012


507 ENDO12L_SAT-479 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) First Reported Case of Adrenocortical Carcinoma and Cushing Syndrome in Patient with Sotos Syndrome Ramya S Vedula, Angela T Boldo, Richard J Comi Dartmouth Hitchcock Medical Center, Lebanon, NH [bold][underline]Background:[/underline][/bold] Sotos syndrome is a rare childhood disorder associated with abnormal facial features, cognitive limitations, behavioral problems, congenital cardiac anomalies, neonatal jaundice, renal anomalies, scoliosis, seizures and tumors. The prevalence of Sotos syndrome is estimated at 1:14,000. The majority of cases are sporadic mutations but autosomal dominant pedigrees are reported. NSD1 (nuclear receptor binding SET domain protein 1) is the only gene associated with Sotos syndrome, it is found in about 80-90%.[br][bold][underline]Case: [/underline][/bold]28 y.o. male with Sotos syndrome presented with weight gain and bilateral pitting edema. Evaluation found dorsal fat pad, large purple striae on abdomen, moon facies, stage II HTN, and hypokalemia. Abdominal MRI showed a right 12cm mass consistent with adrenocortical carcinoma compressing the IVC. Lab studies showed K+ 3.2mmol/L, random cortisol of 44.4mcg/dL, elevated DHEAS of 681mcg/dL and a midnight salivary cortisol of 1380ng/dL with a suppressed ACTH of [lt]5pg/mL. Pt was initially treated with spironolactone and potassium supplementation and was later placed on ketocanazole.[br]Biopsy of the adrenal mass was consistent with adrenocortical carcinoma and stained negative for KRAS, BRAF V600E and EGFR. Pt was also found to have metastatic disease to the lungs with bilateral pulmonary nodules with the largest of 14mm in the right lower lobe and largest on the left 6mm in left upper lobe. Pt was deemed inoperable due to his large tumor burden and metastasis and was started on Mitotane in addition to ketocanozole. He passed away shortly after discharge.[br][bold][underline]Conclusion:[/underline][/bold] Patients with Sotos syndrome are pre-disposed to developing tumors. Although there are reports of tumors in the brain, liver, kidney, bone, jaw, skin, and intestines, this is the first reported case of Cushing[apos]s syndrome and adrenocortical carcinoma in a patient with Sotos syndrome.[br][br]Tatton-Brown K, Cole TRP, Rahman N. Sotos Syndrome. In: Pagon RA, Bird TD, Dolan CR, Stephens K, editors. GeneReviews [Internet]. Seattle (WA): University of Washington, Seattle; 1993-.2004 Dec 17. Leventopoulos G, Kitsiou-Tzeli S, Kritikos K, Psoni S, Mavrou A, Kanavakis E,Fryssira H.A clinical study of Sotos syndrome patients with review of the literature. Pediatr Neurol. 2009 May;40(5):357-64. Cohen MM Jr. Tumors and nontumors in Sotos syndrome. Am J Med Genet. 1999 May 21;84(2):173-5.[br][br]Nothing to Disclose: RSV, ATB, RJC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1291 65 568 SAT-479 PO45-01 Saturday 507 2012


508 ENDO12L_SAT-480 POSTER SESSION: Adrenal Disorders Case Reports I (1:30 PM-3:30 PM) Cortisol, Estrogen (E2) and Androgen-Secreting Adrenal Cortical Carcinoma (ACC) Mohammed NMN Ahmed, Hindi NMN Al-Hindi, Imaduddin NMN Kanaan King Faisal Specialist Hospital [amp] Research Center, Riyadh, Saudi Arabia; King Faisal Specialist Hospital [amp] Research Center, Riyadh, Saudi Arabia; King Faisal Specialist Hospital [amp] Research Center, Riyadh, Saudi Arabia [bold]Background:[/bold][br]We report a case of ACC associated w/high levels of serum cortisol (CS), testosterone [amp] E2. Although not studied specifically in our case, nevertheless, it provides a model for understanding sex-steroid metabolism in ACC. Elevated E2 in pts w/ACC is due to probable dual source of E2 from tumor [amp] peripheral conversion of tumor-derived androgens(1). Aromatase is terminal enzyme responsible for E2 biosyntehesis. High levels of aromatase protein [amp] mRNA have been demonstrated in ACC, but not in adjacent normal tissue(2). Aromatse gene (C19) is regulated in cell-specific manner via alternative use of various promoters in first exon(3). Promoter II mainly directs expression of armoatase gene in all testicular cell types(3). Androgens up-regulate aromatse gene expression in purified adult germ cells(4). The case suggests that utilization of promoter II-directed gonadal-type exon 1 might be involved in over production of aromatase in E2-secreting ACC.[br][bold]Clinical Case[br][/bold]A 27-yr.old man presented w/Cushing[apos]s syndrome(CS) [amp] gynecomastia. Evaluation: consistent w/ACTH-independent CS: elevated 24-hr urinary free cortisol (UFC:3388 nmol, nl,[lt]250),AM serum cortisol:1312 nmol/l, (nl,170-356), ACTH[lt] 1 ng/l,(nl 5-60),E2x2: 45,950 [amp]44, 613 pmol/l,(nl 28-156), testosterone [gt]26 nmol/l x3, (nl,9.9-26), androstenedione [gt]70 nmol/lx3 (nl,2.4-12.6,)17-OH-PX2: 2090[amp]2650ng/dl,(nl,55-455) DHEAS280 ng/ml, (nl,[lt]13). Imaging Studies: CT/MRI/FDG-PETCT:11x10 cm Rt. adrenal tumor w/liver,[amp] IVC invasion, total occlusion of IVC extending cranially within 3 cm rt. atrium, lung mets. FU images: Extensive necrosis w/coincidental drop in E2 to 8,251, UFC to 67 [amp]SC to 339,ACTH increased to 31, all these unrelated to laprotomy for unresectable ACC followed by resurgence of hypercortisolemia (UFC 2297, SC 1231,Drop in ACTH [lt]1). Histopath: Compatible w/ACC w/hi nuclear grade, extensive hemorrhage [amp] necrosis immunostains: +ve for calretnin, inhibin, melan A, AE1/AE3,CAM 5.2. RX: Mitotane, ketoconazole,[amp] Metyrapone tried w/out response, considered not a candidate for Chemo/Radiation Rx.[br][bold]Conclusion:[/bold][br]The case supports the notion that E2-secreting tumors in adult males are frequently carcinomas. Extensive tumor necrosis can result in cyclical steroid hormonal changes. The case provides clinical model for understanding sex-steroid metabolism in ACC, lending support to the concept that high androgenic production by tumor leads to excess E2 synthesis by up-regulating Aromatse gene (C19).[br][br]1)Zayed A, Stock Jl, Liepman MK, Wollin M, Longscope C: Feminization as a result of both peripheral conversion of androgens and direct estrogen production from an adrenal carcinoma. J Endocrinol Invest. 1994;17(4): 275-8. 2)Watanabe T, Yasuda T, Noda H, et al. Estrogen secreting adrenaocarcinoma in an 18-month-old boy: aromatase activity, protein expression, mRNA and utilization of gonadal type promoter. Endocr J. 2000:47(6): 723-30. 3)Carreau S, Bourguiba S, Lambard S, et al. The promoter(s) of aromatase gene in male testicular cells. Reprod Biol. 2004; 4(1):23-34. 4)Bourguiba S, Lambard S, Carreau S. Steroid control the aromatase gene expression in purified germ cells from the adult male rat. J Mol Endocrinol. 2003; 31(1): 83-94.[br][br]Nothing to Disclose: MNMNA, HNMNA-H, INMNK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 330 65 569 SAT-480 PO45-01 Saturday 508 2012


509 ENDO12L_SAT-481 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) miR-449a Is Involved in Glucocorticoid-Induced Downregulation of CRF Type 1 Receptor Expression Takahiro Nemoto, Tamotsu Shibasaki Nippon Medical School, Tokyo, Japan The hormones composing the hypothalamic-pituitary-adrenal (HPA) axis such as corticotropin-releasing factor (CRF), adrenocorticotropic hormone, and glucocorticoids play important roles in stress response. The HPA axis is controlled by the feedback of glucocortioids on the hypothalamus and pituitary. Glucocorticoids reportedly down regulate the expression of not only CRF mRNA in the hypothalamus and proopiomelanocortin mRNA in the anterior pituitary, but also CRF type 1 receptor (CRF-R1) mRNA and protein in the anterior pituitary. However, the mechanism underlying the glucocorticoid-induced CRF-R1 downregulation was not fully understood.[br]miRNAs are short RNA molecules that are found in most types of eukaryotic cells, and are post-transcriptional regulators that usually induce translational repression and gene silencing after binding to the complementary sequence of target mRNAs. We hypothesized that glucocorticoids may induce the expression of miRNAs in the anterior pituitary, which may be involved in glucocorticoid-induced downregulation of CRF-R1.[br]To test this hypothesis, we found 3 miRNAs with sequences predicted to bind to CRF-R1 3[apos] untranslated region (3[apos]UTR) by data base search. Immobilization significantly increased the expression of miR-449a, one of the 3 miRNAs, in the anterior pituitary of rats. The diurnal change of miR-449a expression in the pituitary was in parallel with that of plasma corticosterone. miR-449a expression was significantly increased by dexamethasone, but not by CRF, in the monolayer cultured anterior pituitary cells. Adrenalectomy blocked immobilization-induced increase in miR-449a expression in the anterior pituitary. While overexpression of miR-449a decreased CRF-R1 mRNA and protein expression, knockdown of miR-449a blocked dexamethasone-induced decrease in CRF-R1 mRNA and protein expression in the monolayer-cultured pituitary cells. When cells were co-transfected with luciferase vector, which contained CRF-R1 3[apos]UTR, and miR-449a expression vector, luciferase activity was significantly decreased in HEK 293 cells.[br]These results suggest that miR-449a is involved in glucocorticoid-induced downregulation of CRF-R1 expression.[br][br]Nothing to Disclose: TN, TS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1386 66 570 SAT-481 PO03-02 Saturday 509 2012


510 ENDO12L_SAT-482 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Retinoid X Receptor Agonists Differentially Affect Proliferation, Apoptosis, and ACTH Secretion/POMC Gene Expression in AtT20 Corticotroph Cells Akiko Saito-Hakoda, Akira Uruno, Kyoko Shimizu, Masataka Kudo, Takako Saito-Ito, Takeo Yoshikawa, Ken Matsuda, Ikuma Fujiwara, Hiroyuki Kagechika, Yasumasa Iwasaki, Sadayoshi Ito, Akira Sugawara Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku University Graduate School of Medicine, Sendai, Japan; Tokyo Medical and Dental University, Tokyo, Japan; Kochi University, Kochi, Japan Introduction: Retinoids are possible candidates for the novel therapeutics against adrenocorticotropic hormone (ACTH)-dependent Cushing[apos]s syndrome. In ENDO 2011, we demonstrated the inhibitory effect of retinoid X receptor (RXR) agonist HX630 on ACTH secretion and proopiomelanocortin (POMC) gene expression in AtT20 cells. We here examined the effects of various RXR agonists on proliferation, apoptosis, and ACTH secretion/POMC gene expression. Methods: Mouse pituitary corticotroph AtT20 cells were used. The cells were incubated with RXR agonists (HX630, PA024, or CD3254) for 72 h. Cell proliferation was determined using Cell Counting Kit-8, and cell apoptosis was determined using Homogeneous Caspases Assay. POMC mRNA expression was determined by real-time PCR, and ACTH secretion was measured by ELISA. For POMC gene transcription, a newly obtained stable AtT20 cell line expressing POMC gene 5[apos]-flanking region/luciferase cDNA chimeric construct was used. Results: All RXR agonists induced AtT20 cell apoptosis dose-dependently. HX630 and PA024, but not CD3254, inhibited AtT20 cell proliferation at their high doses. HX630, but not PA024 and CD3254, inhibited POMC gene transcription/mRNA expression and ACTH secretion at its higher doses. Conclusion: Among RXR agonists, HX630 may be the best candidate for the novel therapeutics against ACTH-dependent Cushing[apos]s syndrome.[br][br]Nothing to Disclose: AS-H, AU, KS, MK, TS-I, TY, KM, IF, HK, YI, SI, AS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1434 66 571 SAT-482 PO03-02 Saturday 510 2012


511 ENDO12L_SAT-483 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Impact of Lactation on Cortisol Response to Psychosocial and Predator Stress in Sheep Charlotte Keating, Alan J Tilbrook Monash University, Melbourne, Australia; Monash University, Clayton, Australia Activation of the hypothalamo-pituitary adrenal (HPA) axis by psychosocial stress is commonly attenuated during lactation(1). Evidence in humans suggests that this attenuation may not occur in response to all types of stress(2), particularly when the welfare of the infant may be perceived by the dam to be threatened. We tested this hypothesis in a sheep model using two stressors: psychosocial stress (isolation and restraint) and predator stress (barking dogs). Predator stress would be perceived to threaten the welfare of the lamb. Blood samples were collected i.v. every 10min from non-lactating ewes (n=6), lactating ewes with lambs present but not able to suckle (n=6) and lactating ewes with lambs present and able to suckle (n=6). All animals were exposed to both stressors on different days after 1h of control sampling. Isolation and restraint was imposed for 4h while predator stress was imposed for 5min every h for 4h. For non-lactating ewes there was a significant (P[lt]0.01) increase in cortisol in response to both stressors and these increases were significantly greater (P[lt]0.01) than in both groups of lactating animals. For both groups of lactating ewes there was a significant (P[lt]0.05) increase in cortisol in response to both stressors with a greater response to predator stress (P[lt]0.05). Nevertheless, it is important to note that these increases in cortisol in lactating ewes in response to both stressors were significantly less than those in non-lactating ewes (P[lt]0.01). These data confirm that lactation attenuates HPA axis responses to stress and indicate that the extent of this attenuation is likely to vary with the type of stressor. While the application of the stressors varied, it appears that dams are likely to have a greater HPA axis response to a stressor that is perceived to threaten the welfare of their offspring.[br][br](1) Tilbrook AJ et al., Endocrinology 2006; 147; 3501. (2) Kaye J et al., Clinical Endocrinology 2004; 61; 582.[br][br]Nothing to Disclose: CK, AJT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 356 66 572 SAT-483 PO03-02 Saturday 511 2012


512 ENDO12L_SAT-484 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Oxidative Stress and the Regulation of ALADIN, the Triple A Syndrome Gene Product and Its Interacting Protein Partner Ferritin Heavy Chain Rathi Prasad, Adrian J Clark, Helen L Storr William Harvey Research Institute, Barts and the London School of Medicine, Queen Mary University of London, London, UK [bold]Background[/bold][br]Triple A syndrome is a rare, autosomal recessive cause of adrenal failure that usually manifests in the first decade. Additional features include alacrima ([sim]90%), achalasia of the oesophageal cardia ([sim]75%), and a progressive neurodegenerative process ([sim]60%). The [italic]AAAS[/italic] gene product is the nuclear pore complex protein ALADIN of unknown function. Ferritin Heavy Chain (FTH1), known to have a DNA protective role, is an interacting protein partner with ALADIN(1). Unlike control cells, no nuclear FTH1 is apparent in AAAS fibroblasts implicating ALADIN in the nuclear localisation of FTH1(1). Furthermore apoptosis of neuronal cells induced by hydrogen peroxide is significantly reduced by transfection of [italic]AAAS[/italic] or [italic]FTH1[/italic] and maximally by both genes together(1), implicating oxidative stress in the pathogenesis of the disease.[br][bold]Aim[/bold][br]To investigate the expression of ALADIN and FTH1 in conditions of oxidative stress in H295R adrenocortical tumour cells and SH-SY5Y neuroblastoma cells (chosen as representative of the cell types affected by AAAS).[br][bold]Methods[/bold][br]Both cell lines were treated with hydrogen peroxide and mRNA expression of [italic]AAAS[/italic] and [italic]FTH1[/italic] was quantified over a 24 hour period, using real time qPCR. Protein expression at 24 hours was quantified by densitometric analysis following immunoblotting.[br][bold]Results[/bold][br][italic]FTH1[/italic] mRNA expression was significantly increased following treatment with a peak response at 6 hours in H295R cells (mean increase of 135%, n=3, p[lt]0.05) and SH-SY5Y cells (mean increase of 96%, n=3, p[lt]0.001). However in SH-SY5Y cells, [italic]AAAS[/italic] mRNA expression was significantly reduced in response to oxidative stress (mean reduction of 56%, n=3, p[lt]0.05). Indeed 24 hours after treatment protein expression of ALADIN in SH-SY5Y cells is significantly reduced (mean reduction of 46%, n=3, p[lt]0.005). In comparison, [italic]AAAS[/italic] mRNA expression and ALADIN protein expression are maintained in H295R cells with treatment.[br][bold]Conclusion[/bold][br]The adrenal cortex and neural tissue are highly oxidative environments. FTH1 has potent ferroxidase activity and its mRNA expression is upregulated in both cell lines following application of oxidative stress. Interestingly, ALADIN appears to be downregulated both at transcriptional and translational levels in SH-SY5Y cells in response to oxidative stress whereas levels are maintained in H295R cells. Further exploration of the mechanisms of this differential regulation may have implications for susceptibility of these tissues to oxidative stress in other disease states.[br][br](1) Storr HL et al., Molecular Endocrinology 2009; 12: 2086-2094.[br][br]Nothing to Disclose: RP, AJC, HLS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1536 66 573 SAT-484 PO03-02 Saturday 512 2012


513 ENDO12L_SAT-485 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Gender-Independent Control of Adrenocorticotropic Hormone (ACTH) Secretion under a Varying Cortisol Infusion Clamp in Older Men and Older Women Animesh N Sharma, Paul Aoun, Sue Weist, Jean Wigham, Johannes D Veldhuis Mayo Clinic, Rochester, MN [bold]Introduction:[/bold][br]Stress-adaptive responses of the corticotropic axis are critical to sustain life and longevity. Either impaired feedback repression by elevated cortisol or accentuated rebound from a low cortisol could mediate altered stress adaptations. Our objective is to discern gender associated ACTH secretory responses to feedback enhancement in older men and women. We also appraise the role of gender in determining the regularity of ACTH secretion as well as the integrity of the feedback system via approximate entropy (ApEn) and Cross ApEn (X-ApEn) respectively[sup]1-3[/sup].[br][bold]Methods/Design:[/bold][br]A prospectively randomized double-blinded, placebo controlled, crossover study, conducted in 10 older men and 10 older women. During four separate hospital visits, each patient received oral placebo followed by intravenous saline (IVS), or oral ketoconazole (KTCZ) followed by an infusion of either IVS, low dose cortisol (LDC), or high dose cortisol (HDC). Blood sampling was performed every 10 minutes for ACTH and cortisol from 10 pm to 2 am, at which point the infusion was discontinued.[br][bold]Result:[/bold][br]There was no difference in mean ACTH concentration, ACTH ApEn and feedback X-ApEn with regards to gender. Gender independence was confirmed by insignificant difference in feedback slope (p=0.45) in a model that showed infusion-dependent suppression. There was also no significant difference in inter-individual variability (standard deviation {SD}) for mean ACTH, ACTH ApEn and feedback X-ApEn between gender types.[br][bold]Conclusion:[/bold][br]While experimental hypercortisolemia significantly suppresses ACTH release and enhances feedback regularity in older individuals, gender does not govern the foregoing parameters. While these findings may suggest a lack of inter-gender variability in ACTH secretion and feedback control, they should be contrasted with similar gender related studies in younger individuals.[br][br](1)Veldhuis, J.D. and S.M. Pincus, European journal of endocrinology / European Federation of Endocrine Societies, 1998. 138(4): p. 358-62. (2)Pincus, S.M., Novartis Foundation symposium, 2000. 227: p. 82-96; discussion 96-104. (3)Pincus, S.M., et al., The American journal of physiology, 1999. 277(5 Pt 1): p. E948-57.[br][br]Sources of Research Support: NIH Grant DK73148 awarded to Dr. J.D.Veldhuis.[br][br]Nothing to Disclose: ANS, PA, SW, JW, JDV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1105 66 574 SAT-485 PO03-02 Saturday 513 2012


514 ENDO12L_SAT-486 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Diminished ACTH Sensitivity and Efficacy in Obese Premenopausal Women Ferdinand Roelfsema, Hanno Pijl, Daniel M Keenan, Johannes D Veldhuis Leiden University Medical Center, Leiden, Netherlands; University of Virginia, Charlottesville, VA; Mayo Clinic, Rochester, MN Obesity is associated with various neuroendocrine perturbations (i.e. ACTH, PRL, TSH, GH) and peripheral hormone abnormalities (insulin, adipokines).We previously demonstrated the activated ACTH-cortisol axis in obese premenopausal women associated with decreased ACTH potency (1,2). Recently, a new method for constructing the endogenous dose-response relationship was introduced (3), which is based on the relation between ACTH concentrations and associated cortisol secretion rates within cortisol bursts. We retrospectively analyzed ACTH-cortisol profiles in 25 obese premenopausal women and 15 lean premenopausal women. We used the 2-potency model in which the initial potency is not necessarily equal to that during recovery phase (4).The initial potency (negative logarithm) was -7.83 [plusmn] 0.75 (mean [plusmn] SEM) in obese women, and -10.14 [plusmn] 1.08 in lean women (P=0.10) and the corresponding values for the recovery phase were -26.62 [plusmn] 2,21 and -36.67 [plusmn] 1.66 (P = 0.004).The sensitivity (curve slope)amounted 0.468 [plusmn] 0.05 in obese women and 0.784 [plusmn] 0.09 in lean women (P=0.004).The efficacy (maximal value) was 17.6 [plusmn] 4.9 nmol/L.min in obese women and 26.3 [plusmn] 3.8 nmol/L.min in lean women (P = 0.01). No differences were present for basal secretion rate, inflection point and EC[sub]50 [/sub]values. The ACTH-cortisol relation in obesity in women is characterized by decreased sensitivity and efficacy, thus explaining non-elevated serum cortisol concentrations in view of increased plasma ACTH levels. The relation described here differs markedly from that found in Cushing[apos]s disease (5). In the latter condition, the dose-response curve is shifted to the right (increased EC[sub]50[/sub]) with increased efficacy, which normalizes after curative surgery. The present results only apply to premenopausal obese women. Whether a similar dose-response curve is present in men and in postmenopausal women is not known yet. The mechanisms behind the changed dose-response curve are not known, as well as whether this relation can be repaired after weight reduction. Increased secretion of ACTH [italic]per se[/italic] is not a causal mechanism, because short term treatment with bromocripine or Acipimox, which restores excessive ACTH secretion to normal values, does not change the dose-response curve.[br][br](1) Kok et al., J Clin Endocrinol Metab 2004;287:848. (2) Roelfsema et al.,J Clin Endocrinol Metab 2009;94:2991. (3) Chattopadhyay et al., Quart Appl Mathem 2008;66:401. (4) Keenan et al., Am J Physiol 2010;299:R11. (5) Roelfsema et al.,J Clin Endocrinol Metab 2011;96:3768.[br][br]Nothing to Disclose: FR, HP, DMK, JDV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 48 66 575 SAT-486 PO03-02 Saturday 514 2012


515 ENDO12L_SAT-487 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Normal Ranges of Basal and ACTH-Stimulated Free Cortisol in Children Ori Eyal, Naomi Weintrob, Irina Serdukov, Asaf Oren, Tali Taxir, Anita Schachter Davidov, Naftali Stern, Rona Limor Dana-Dwek Children[apos]s Hospital, Tel Aviv Sourasky Medical Center, Tel Aviv, Israel; Tel Aviv University, Tel Aviv, Israel; Tel Aviv Sourasky Medical Center, Tel Aviv, Israel [bold]Background[/bold]: Standard assays for serum cortisol measurements determine total cortisol (TC) concentrations but not the unbound biologically active serum free cortisol (sFC). Measurement of TC would be greatly influenced by alteration in cortisol-binding globulin (CBG) concentrations. It is, therefore, important to determine sFC levels when CBG levels are either decreased or increased.[br][bold]Objectives[/bold]: To determine basal and ACTH-stimulated sFC levels in healthy children and to assess their relationship to TC, age, gender and Tanner stage.[br][bold]Subjects and methods[/bold]: Baseline and stimulated serum TC and FC concentrations were measured before and after IV administration of 250 mcg Synacthen (Defiante Farmaceutica, S.A., Portugal) in healthy children referred for exclusion of adrenal dysfunction. Serum TC was determined by chemiluminescence (Rosh, Cobas A 411), and serum FC was measured by the same methods following equilibrium dialysis. A TC response of 20 mcg/dl was considered normal.[br][bold]Results[/bold]: The study group consisted of 55 subjects (M/F 17/38) whose median age was 8.5 years (range, 0.6-16.9). Mean baseline TC and sFC levels (mcg/dl, 95 CI) were 11.7 (10.0-13.3) and 0.35 (0.27-0.42), respectively. Mean peak TC and sFC levels were 31 (30.5-33.3) and 1.6 [plusmn] 0.5 (1.46-1.73), respectively. Mean fractions of sFC at baseline and at peak were 3.1 % (2.8-3.4) and 5.88 % (5.4-6.3) (p[lt]0.001), reflecting a lower increase in TC (247%) compared to sFC (647%). Baseline and peak TC and sFC levels were positively correlated (r=0.81, p[lt]0.001 and r=0.43, p[lt]0.001, respectively). Peak TC levels and age were negatively correlated (r=-0.38, p=0.005). There was a significant difference between TC at baseline and at peak between children aged [lt]10 years compared to those [gt]10 years (p=0.04 and 0.01, respectively).[br][bold]Summary[/bold]: Based on these findings, we suggest normal ranges for basal and ACTH-stimulated sFC. The finding that TC, but not sFC, is age-dependent indicates that sFC may be more reliable than TC for measuring adrenal reserve in children.[br][br]Nothing to Disclose: OE, NW, IS, AO, TT, ASD, NS, RL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 325 66 576 SAT-487 PO03-02 Saturday 515 2012


516 ENDO12L_SAT-488 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) The Role of Basal Serum Cortisol in the Assessment of Hypothalamic Pituitary Adrenal Function in Children Arati Mokashi, Fatima Imran, Sam Stewart, Lisa Tramble, David Clarke, Stephanie M Kaiser, Mohammad Alotaibi, Syed Ali Imran Dalhousie University, Halifax, Canada; Dalhousie University, Halifax, Canada; Dalhousie University, Halifax, Canada; Dalhousie University, Halifax, Canada The data on the role of morning (basal) serum cortisol as a marker of hypothalamic pituitary adrenal (HPA) function in children are mostly derived from the adult literature. The cut-off values which would avoid the need for dynamic testing vary from 80 - 138 nmol/L (lower) to 275 - 525 nmol/L (upper) [1,2]. In our centre, we use lower and upper cut-off values of [lt]80 nmol/L and [gt]275 nmol/L, respectively based on a review by Frank et al [1]. For dynamic testing, we use the low dose (1 microgram) ACTH test as an alternative to the insulin tolerance test which is not recommended in children. For this study, we analyzed consecutive patients since 2009 who had been assessed through the endocrinology clinic for a provisional diagnosis of adrenal insufficiency. Based on the above criteria, patients who had a basal (0800) serum cortisol followed by an ACTH test were identified and their data were analyzed. A total of 46 patients (21 females) underwent testing; the primary diagnoses were prior chronic systemic glucocorticoid therapy (17), asthma/inhaled glucocorticoid therapy (8), congenital hypopituitarism (8), cranial radiation therapy (5), fatigue (3), Prader- Willi syndrome (1), cortisol secreting adrenal tumor post resection (1), hypoglycemia (1) and low basal cortisol of unknown etiology (1). A 30 or 60-minute post-ACTH cortisol peak of [gt]500nmol/L were used as standards to exclude HPA insufficiency. Based on post ACTH peaks of [gt]500 or [gt]550 nmol/L, the lower cut-off values for 0800 basal cortisol providing 100% sensitivity were 112 and 129 nmol/L, respectively, while the upper cut-off values providing 100% specificity were 330 and 350 nmol/L, respectively. Optimal values of serum cortisol providing the best fit were calculated through an ROC analysis. An 0800 serum cortisol value of 233 nmol/L while using a post-ACTH peak of [gt]500 nmol/L provided a sensitivity of 70% and specificity of 68%, whereas an 0800 cortisol of 203 nmol/L while using a post-ACTH peak of [gt]550 nmol/L provided a sensitivity of 77% and a specificity of 58%. In conclusion, our data suggest that an 0800 serum cortisol may be used as an alternative to dynamic testing in children.[br][br](1) Frank GR et al. Safety of Medications and Hormones Used in Pediatric Endocrinology: Adrenal. Ped. Endocrinol. Rev. 2004; 2(suppl 1):134-145. (2) Oelkers W. Adrenal Insufficiency. N Eng. J. Med. 1996; 355:1206-1212.[br][br]Nothing to Disclose: AM, FI, SS, LT, DC, SMK, MA, SAI 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 570 66 577 SAT-488 PO03-02 Saturday 516 2012


517 ENDO12L_SAT-489 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Distinct Effects of Macronutrients on Post-Prandial Adrenal and Extra-Adrenal Cortisol Production in Humans Roland H Stimson, Nor A Mohd-Shukri, Ruth Andrew, Jennifer L Bolton, Rebecca M Reynolds, Brian R Walker University of Edinburgh, Edinburgh, UK Cortisol secretion and metabolism are affected by chronic energy imbalance (starvation or obesity), and dietary macronutrient content may modify these effects (e.g. a low carbohydrate (CHO) diet increases hepatic cortisol regeneration by 11[beta]-HSD1). Circulating cortisol concentrations also increase post-prandially, but it is unclear whether this is specific to certain macronutrients and reflects increased adrenal secretion, increased regeneration by 11[beta]-hydroxysteroid dehydrogenase type 1 (11[beta]-HSD1), or reduced cortisol clearance. We used similarly flavoured liquid meals to test the effects of individual macronutrients on cortisol secretion and metabolism, measured by stable isotope tracer infusion.[br]Eight normal weight healthy men attended after overnight fast on 4 occasions separated by [gt]1 week. At each visit, 9,11,12,12-[[sup]2[/sup]H][sub]4[/sub]-cortisol (d4-cortisol) was infused to steady state (t+3.5 h); subjects were given liquid meals composed of either CHO, protein, fat ([sim]560 kcal/meal), or low calorie placebo in a randomised double-blind crossover design; and tracer infusion and blood sampling continued for 3 h. Local ethical approval was obtained.[br]Plasma cortisol, the rate of appearance (Ra) of cortisol (a composite of adrenal secretion and extra-adrenal regeneration), Ra d3-cortisol (a specific measure of 11[beta]-HSD1 regeneration), and d4-cortisol clearance were similar before each meal. Plasma cortisol and total Ra cortisol were increased similarly by all meals compared with placebo (Ra cortisol by 84% after CHO, 96% after protein, and 43% after fat, all p[lt]0.05). In contrast, Ra d3-cortisol was increased only by CHO and protein (by 47% and 38%, respectively, p[lt]0.05). d4-Cortisol clearance was unchanged following CHO and fat, but increased by protein (by 21%, p[lt]0.05). Plasma insulin was increased markedly by CHO (p[lt]0.001) and slightly by protein (p[lt]0.05).[br]All classes of dietary macronutrient increase adrenal cortisol secretion. However, while a high fat meal has no effect on cortisol metabolism or clearance, both CHO and protein increase regeneration of cortisol by 11[beta]-HSD1, putatively via increased insulin. The lack of reduction in cortisol clearance suggests that CHO and protein also increase activity of cortisol-inactivating enzymes. These novel findings suggest that post-prandial tissue glucocorticoid levels vary substantially with different diets, potentially contributing to deleterious effects of, for example, high glycemic index foods.[br][br]Sources of Research Support: This work was funded by the British Heart Foundation.[br][br]Nothing to Disclose: RHS, NAM-S, RA, JLB, RMR, BRW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 911 66 578 SAT-489 PO03-02 Saturday 517 2012


518 ENDO12L_SAT-490 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Do Children and Young Adults with Prader-Willi Syndrome (PWS) Have an Increased Prevalence of Central Adrenocortical Insufficiency? Berthold P Hauffa, Katja Konrad, Corinna Grasemann, Christian Dohna-Schwake, Eva Tschiedel, Nicole Unger, Dagmar Fuhrer-Sakel, Christiane Maser-Gluth, Stephan Petersenn University Children[apos]s Hospital, Essen, Germany; University Hospital, Essen, Germany; University of Heidelberg, Heidelberg, Germany; Outpatient Clinic for Endocrinology, Andrology and Medical Oncology, Hamburg, Germany [bold]Introduction:[/bold] The annual death rate in children with Prader-Willi syndrome (PWS) is elevated (3 %), sudden and unexpected deaths do occur. This has been attributed in part to dysfunction of the corticotropic axis at the hypothalamic level. Using a metyrapone test, 60 % of PWS children (1) were found to have central adrenal insufficiency (CAI), and hydrocortisone replacement during stressful conditions has been recommended. Fewer children ([lt] 15 %) were identified with the low dose ACTH test (LDAT) (2). No cases of CAI were found in PWS patients (3) when high dose ACTH testing (HDAT) or insulin hypoglycaemia (IHT) were used. Metyrapone test results of other PWS cohorts have not been reported. [bold]Patients and Methods:[/bold] We therefore examined 25 children with PWS (age 8.1 [plusmn] 5.6 yrs [mean [plusmn] 1 SD]), performing a single dose overnight metyrapone test. At 11:30 PM 29.1 [plusmn] 2.7 mg/kg metyrapone was given po. Blood samples for measurement of cortisol, 11-desoxycortisol, ACTH and glucose were obtained at 4:00 AM, 6:00 AM, and 7:30 AM. An ACTH increase [gt] 150 pg/ml was considered sufficient to exclude CAI. Seven young adult PWS patients (age 17.5-20.8 years) had the corticotropic axis evaluated by IHT during workup for adult growth hormone deficiency. A rise of serum cortisol [gt] 500 nmol/l in response to a fall in blood glucose of [lt] 2.2 mmol/l was considered sufficient to exclude CAI. [bold]Results:[/bold] Over the past years, 10/25 PWS children had slightly decreased basal AM cortisol concentrations on 1-4 occasions, the lowest cortisol concentration being 59 nmol/l. None of the children had clinical evidence of adrenal insufficiency during acute illness. Pre-metyrapone cortisol (ACTH, 11-desoxycortisol) was 187 [plusmn] 156 nmol/l (18.3 [plusmn] 14.8 pg/ml, 0.31 [plusmn] 0.19 ng/ml). After metyrapone, cortisol decreased to 50.7 [plusmn] 25.0 nmol/l, ACTH (11-desoxycortisol) increased to 278 [plusmn] 206 pg/ml (13.1 [plusmn] 4.8 ng/ml). Blood glucose did not change, blood pressure and heart rate remained stable. Six of 25 children (25 %) failed to pass the metyrapone cut-off and fulfilled the criteria for CAI. None of the young adult PWS patients failed to increase serum cortisol above 500 nmol/l. [bold]Conclusion:[/bold] In PWS, CAI diagnosed by an insufficient rise in ACTH after a single night-time metyrapone dose may be less frequent than originally thought. Differences between studies may depend on ACTH assays, pre-analytical conditions and stimuli used. The risk of a PWS child to progress into CAI with age appears to be negligible.[br][br](1)De Lind van Wijngaarden RFA et al., JCEM 2008; 93:1649. (2)Corrias A et al., Clin Endocrinol 2012, doi:10.1111. (3)Farholt S et al., JCEM 2011; 96:E173.[br][br]Nothing to Disclose: BPH, KK, CG, CD-S, ET, NU, DF-S, CM-G, SP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2147 66 579 SAT-490 PO03-02 Saturday 518 2012


519 ENDO12L_SAT-491 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Urocortin System: A Link between Heart and Renal Diseases Shilpi Mahajan, Aditi Bhargava University of California San Francisco (UCSF), San Francisco, CA Renal disease is the strongest risk factor for cardiovascular morbidity and mortality. Chronic kidney disease (CKD) is a major contributor to severe cardiac damage and, conversely, congestive heart failure is a major cause of progressive CKD. Urocortins (Ucns), members of the corticotrophin-releasing factor (CRF) family, are important mediators of stress and immune function. Their cardiovascular effects are mediated via the CRF receptors CRF[sub]1[/sub] and CRF[sub]2[/sub] through Ca[sup]2+[/sup] or cAMP intra-cellular signaling pathways. Reports conflict regarding Ucn2[apos]s role in regulating the aldosterone (Aldo) and plasma Na[sup]+[/sup] level during heart failure. The mechanistic links between Ucn, heart failure and kidney diseases have not been established. Here we show by RT-PCR that Ucn2, CRF[sub]1[/sub], CRF[sub]2[/sub], and components involved in mediating Ucn2 signaling are expressed in kidneys under basal conditions and are regulated after Aldo treatment. Aldo increased Ucn2 levels 3-fold over baseline in total kidney and in cortical collecting ducts (CCD). CRF[sub]1[/sub] was expressed in total kidney, but not in CCD and did not respond to Aldo treatment. CRF[sub]2[/sub] was undetectable in total kidney but robustly increased in CCD cells after dexamethasone treatment. Therefore, CRF[sub]2[/sub] seems to be focally expressed in CCD and regulated in Aldo-responsive areas. We found an inverse relationship between Ucn2 and expression of endothelin-converting enzyme 1 (ECE-1) in response to Aldo treatment in total kidneys. ECE-1 regulates recycling and re-sensitization of some GPCRs by degrading their agonists in endosomes, thereby freeing the receptor to recycle to the cell surface, which mediates re-sensitization. HPLC data showed that ECE-1 differentially degrades Ucn1-3. ECE isoforms (b, c and d) were down-regulated in response to Aldo treatment in total kidney. In contrast, in CCD cells, ECE-1 b [amp] c isoforms were upregulated in response to dexamethasone. Thus, ECE1 actions appear important in regulating the expression of Ucn2 in kidney and Ucn2 might be critical in decreasing Aldo level to normal, alleviating hypertension. Ucn2 stimulation resulted in CRF[sub]2[/sub] trafficking from the plasma membrane into cytosolic vesicles as seen under confocal microscopy, and stimulation of HEK-CRF[sub]2[/sub] cells with Ucn2 evoked a significant intracellular Ca[sup]2+[/sup] response. We therefore propose a model in which the Ucn system is the link that mediates its pleiotropic effects on heart and kidney during stress and inflammation via tight regulation of its receptor activation.[br][br]Sources of Research Support: R01 NIDDK/NIH 3DK080787-01 grant to AB.[br][br]Nothing to Disclose: SM, AB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 112 66 580 SAT-491 PO03-02 Saturday 519 2012


520 ENDO12L_SAT-492 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) The Role of Pattern Recognition Receptors and Related Inflammation in Adrenal Gland Dysfunction during Sepsis Waldemar Kanczkowski, Nguyen Tran, Vanessa Hoesker, Uta Lehnert, Monika Ehrhart-Bornstein, Triantafyllos Chavakis, Kai Zacharowski, Stefan Bornstein Technical University Dresden, Dresden, Germany; University Hospital Frankfurt, Frankfurt am Main, Germany Sepsis and septic shock in response to bacterial or viral infections remain a major health care problem worldwide. Patients with diagnosed adrenal gland insufficiency are at high risk of developing a multi-organ dysfunction syndrome which dramatically increases risk of mortality. However, little is known about the mechanisms leading to improper adrenal function in these conditions.[br]In the present study, we tested our hypothesis that sepsis-related activation of the innate immunity pattern recognition receptors (PRRs) increases adrenocortical cell inflammation, oxidative stress, and cell death thereby contributing to their impaired function.[br]Using real-time PCR, we compared gene expression in the adrenal glands obtained from mice with polymicrobial sepsis (cecal ligation and puncture model) plus appropriate sham-operated controls and from mice with systemic inflammation induced by Gram-positive and -negative bacterial ligands and corresponding saline-treated animals. Our results showed a pronounced changes in expression of genes encoding for PRRs such as toll-like receptors (TLR2, 4), NOD-like receptors (NLRP3, NOD1 and NOD2) and PRR-related inflammasome components (ASC, caspase 1). Furthermore, we observed an upregulation of genes involved in oxidative stress, mainly subcomponents of NADPH oxidases (p40phox, p47phox), nitric oxide synthase (iNOS), and proinflammatory cytokines (IL1 beta, TNF alpha, IL6). In addition, we showed increased protein levels of corresponding proinflammatory cytokines using a multiplex ELISA technique. Finally, using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, we identified a dramatic increase in apoptosis rate in adrenal glands obtained from mice undergoing polymicrobial sepsis or with bacterial ligand-induced systemic inflammation as compared to control animals.[br]Taken together, sepsis-induced activation of the innate immunity receptors may contribute to adrenal insufficiency by enhancing tissue inflammation, oxidative stress and culminate in cellular apoptosis.[br][br]Nothing to Disclose: WK, NT, VH, UL, ME-B, TC, KZ, SB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 962 66 581 SAT-492 PO03-02 Saturday 520 2012


521 ENDO12L_SAT-493 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Adrenal Function at Different Stages of Chronic Renal Disease Alejandro Luis Arregger, Estela Maria Lujan Cardoso, Enrique Dorado, Alfredo Zucchini, Alicia Elbert, Rocio Sanchez, Olga Beatriz Sandoval, Liliana Noemi Contreras University of Buenos Aires, Buenos Aires, Argentina; IDIM-CONICET, Buenos Aires, Argentina; University of Buenos Aires, Buenos Aires, Argentina Excess or deficiency of corticoadrenal steroid secretion affect quality of life and is associated to early mortality in general population. Multiple endocrine disturbances are seen in patients with end-stage renal disease and its diagnosis is a challenge due to pitfalls in biochemical testing interpretation. This study aims to validate the accuracy of first line tests used to rule out adrenal dysfunction in patients with chronic kidney disease in stages (S) 1 to 4 (K/DOQI CKD). We screened 34 adults (aged 18-72 yo, 25 women and 9 men) staged: S1:n=14; S2: n=12; S3:N=5; S4: n=3. Thirteen patients had primary kidney disease, fourteen had HTA, three had Diabetes Mellitus type 2, four had obesity. Glomerular filtration rate (GFR ml/min/1.73m[sup]2[/sup]) was stable in all the cases for al least three months previous this study. None were on steroids or drugs that may affect adrenal function. All patients gave their consent to participate in the study. Whole salivary samples were obtained by each patient at 08 h and 23 h in 2 non-consecutive days for salivary cortisol assessment (SAF [sub]8 [/sub] and SAF [sub]23[/sub], respectively). When SAF [sub]8[/sub] was less than 3.0 nM, low-dose ACTH stimulation test (25.0 [micro]g ACTH im) was performed to assess rapid cortisol response. At SAF[sub]8 [/sub]levels[ge] 3.0 nM the fast feed back was assessed through the overnight 1 mg dexamethasone suppression test. Patients with GFR[ge] 60 (n=28) collected 24 hr urine samples for urinary free cortisol (UFC).Reference values: (range) SAF[sub]8 [/sub]= 4.4-18.0 nM; SAF[sub]23[/sub]=0.5-3.8 nM; SAF[sub]dex[/sub] = 0.5-2.0 nM, F[sub]dex[/sub] =13.5- 50.0 nM, UFC: 27.0-248.0 nM, SAF 30 minutes post ACTH = 20.0-60.0 nM.[br]Results: age correlated negatively and significantly with GRF (r= -0.360; p= 0,038) SAF [sub]23[/sub] (mean [plusmn]SD): 1.32 [plusmn] 0.87 nM; SAF[sub]8[/sub]: 8.38 [plusmn]4.9 nM; SAF [sub]dex:[/sub] 1.15 [plusmn] 1.19 nM; F[sub]dex[/sub]: 39.0 [plusmn] 54.0 nM; UFC: 98.0 [plusmn] 47.0 nM/day.SAF [sub]dex[/sub] and F[sub]dex [/sub]showed significant correlation with GFR (r= -0.556 and r= -0.481, respectively; p[lt]0.008 for both). 30/32 patients suppressed normally. In 2 cses suppression was achieved after 2 mg dex. 2/34 patients had SAF[sub]8[/sub] [lt]3.0 nM. One patient accepted an ACTH stimulation test, that confirmed an adequate SAF response (55.0 nM)[br]Conclusion: Patients on S1 did not show false positive results. At difference, variable data were obtained in S2 to S4. In these stages further and cost consuming tests were required to rule out adrenal dysfunction.[br][br]Nothing to Disclose: ALA, EMLC, ED, AZ, AE, RS, OBS, LNC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 190 66 582 SAT-493 PO03-02 Saturday 521 2012


522 ENDO12L_SAT-494 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Pasireotide, Octreotide and Cabergoline Do Not Decrease Basal and ACTH-Stimulated Cortisol Secretion by Primary Cultures of Human Adrenal Glands Rob van der Pas, Richard Feelders, Casper van Eijck, Peter van Koetsveld, Francien van Nederveen, Leo Hofland Erasmus Medical Center, Rotterdam, Netherlands; Erasmus Medical Center, Rotterdam, Netherlands; Erasmus Medical Center, Rotterdam, Netherlands [bold]Introduction[/bold][br]Although transsphenoidal surgery is still the primary treatment for Cushing[apos]s disease (CD), there is a growing body of evidence that shows effectiveness of medical therapy with pasireotide, which targets the somatostatin receptor subtypes (sst) 2 and 5, and cabergoline, which targets the dopamine 2 receptor (D2R), in a subset of patients with CD. Whereas the inhibitory effects of cabergoline and pasireotide on ACTH secretion by pituitary adenomas are well established, their direct effects on cortisol secretion by adrenocortical cells have thus far not extensively been investigated. Previous studies already showed that sst and D2R are expressed in the adrenal cortex.[br]The objective of this study was to evaluate the effects of somatostatin analogs and dopamine agonists on cortisol production by primary cultures of human adrenocortical cells.[br][bold]Methods[/bold][br]Primary cultures of both normal adrenal glands (n=3) and ACTH-dependent adrenocortical hyperplasias (n=5) were incubated with pasireotide, octreotide and cabergoline (all at 10nM). Basal (72h) and ACTH-stimulated (250 pg/mL; 2h) supernatant cortisol concentrations were measured.[br][bold]Results[/bold][br]After 2h, neither drug significantly influenced ACTH-stimulated cortisol secretion by cultured cells of either normal adrenal glands or ACTH-dependent hyperplasias. In addition, a 72h incubation with octreotide or pasireotide did not inhibit basal cortisol secretion. Compared to control, cabergoline increased basal cortisol secretion from normal adrenocortical cells by 28% after 72h (p[lt]0.01). Finally, after a 72h pre-incubation, ACTH-stimulated cortisol secretion was not affected by pasireotide and octreotide. A 72h pre-incubation, followed by a 2h incubation with cabergoline in the presence of ACTH, however, induced a 34% increase in cortisol secretion from cultured cells of ACTH-dependent adrenocortical hyperplasias (p[lt]0.05).[br][bold]Conclusion[/bold][br]Whereas pasireotide, octreotide and cabergoline have been shown to inhibit ACTH secretion by corticotroph pituitary adenomas in patients with CD and Nelson[apos]s syndrome, respectively, these drugs do not seem to directly inhibit adrenocortical cortisol production. Thus, the beneficial effects observed in patients with CD are primarily attributed to their ACTH-lowering effect.[br][br]Nothing to Disclose: RvdP, RF, CvE, PvK, FvN, LH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 916 66 583 SAT-494 PO03-02 Saturday 522 2012


523 ENDO12L_SAT-495 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Usefulness of the Ratio of the Cortisol Concentrations (Concentrations at the End/Concentrations at the Start of the Working) in Evaluating the Night Shift Work on the Circadian Rhythm Fumihisa Miyauchi Ehime Rosai Hospital, Niihama, Japan Introduction[br]The previous study showed that the cortisol concentrations in plasma were affected by the night shift work. To evaluate the effects of working on the circadian rhythm of cortisol, plasma concentrations were compared based on the working conditions, i.e., daytime shift, evening shift, and night shift.[br]Materials and Methods[br]Fifty-one female nurses and twenty-one male nurses who enrolled voluntarily in this study were healthy and non-smokers. They worked on three different shifts from 8:00 to 17:00 (daytime shift), from 17:00 to 24:00 (evening shift), and from 24:00 to 8:00 (night shift). Blood were obtained from each nurse at the start and the end of their shifts. The effects of the shift works were evaluated by comparing cortisol concentrations of plasma obtained every two hours from 8:00 for 24hours and at 17:00 from off-duty nurses (6 female and 5 male). The data from the off-duty nurses were served as a control group to see the circadian rhythm. The effects of shift works were also evaluated by calculating the ratio of the cortisol concentrations (concentrations at the end/concentrations at the start of the working). Concentrations of cortisol were determined by LC-MS/MS.[br]Results[br]The cortisol concentrations at both the start and the end of the daytime shift, and the evening shift in plasma did not differ from those of the control group. Ratio of the daytime shift and the evening shift were not differ with that of the control group. The ratio was significantly decreased in the female group only, although the cortisol concentrations at both the start and the end of the first day of the night shift did not differ from those of the control group of neither the female nor the male groups. In the female group, the cortisol concentrations at the start of the second day of the night shift were significantly elevated and the ratio was also significantly decreased. On the contrarily, plasma concentrations and ratio were similar with those of the control group, respectively, on the second day of the night shift in male group.[br]Conclusions[br]The circadian rhythm of cortisol is maintained during the daytime shift and the evening shift, but affected by the night shift work in the female group. To evaluate the effect of working at night, calculating the ratio (concentrations at the end/concentrations at the start of the work) is more sensitive rather than comparing the examined values of the plasma concentrations.[br][br]Sources of Research Support: This study is supported by Japan Labour Health and Welfare Organization.[br][br]Nothing to Disclose: FM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 589 66 584 SAT-495 PO03-02 Saturday 523 2012


524 ENDO12L_SAT-496 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Is the Urinary Free Cortisol a Reliable Test for Screening of Hypercortisolemia? Maria Castillo Tous, Ainara Madrazo-Atutxa, Juan Francisco Martin-Rodriguez, Antonio Leon-Justel, Miguel Angel Mangas-Cruz, Alfonso Leal-Cerro IBIS/HUVR, Seville, Spain; IBIS/HUVR, Seville, Spain Introduction:[br]Cushing[apos]s syndrome (CS) results from prolonged and inappropriate high exposure of tissues to glucocorticoids. The clinical suspicion of CS always needed confirmation by biochemical tests; 24-h urinary free cortisol (UFC) have been used extensively at first line test for screening of CS. The practical issues that accompany 24-h urine collection, the varied of measurement methods, and the increased of prevalence of mild, preclinical or cyclic CS with the fact cortisol is not uniformly secreted during the day, questioning the value of UFC as screening test for CS. We have compared the efficiency of different test proposed for initial screening of CS.[br]Methods:[br]The study participants were patients under the suspicion of having CS and patients that were referred to our institution (tertiary universitary hospital) to manage proven CS (13 Cushing[apos]s disease, 3 adrenal adenoma and 1 ectopic corticotrophin syndrome). In all patients we measured UFC (three different samples), NSC, and overnight DST. We compared the diagnostic efficiency (sensibility (SB), specificity (SP), positive predictive value (PPV), and negative predictive value (NPV)) of the different test proposed for screening of CS: UFC, NSC, overnight DST.[br]Results:[br]Single test: UFC: SB: 75%%, SP: 97,9%, PPV: 92,3%, NPV:92,2% (three samples out of rage); NPV: 93%; NSC: SB: 88,2%, SP: 76,2%, PPV: 50%. NPV: 96%; DST: SB: 88%, SP: 76%, PPV: 50%, NPV:96%. Test ombinations: UFC + DST: SB: 70,6%, SP: 96,9%, PPV: 85,7%. NPV: 92,5%: NSC + DST: SB: 70,6%, SP: 98,4%, PPV: 92,3%. NPV: 92,5%.[br]Conclusions:[br]No one of the tests studied, when are used as single measurement, have acceptable accuracy for screening of CS.[br]The practical issues that accompany UFC collection, measurement and the necessity for careful patient compliance don[apos]t make its use simply for screening test.[br]NSC has similar diagnostic efficiency that UFC when we use that, either as a single test or in combination with overnight DST for the screening of CH.[br][br]Nothing to Disclose: MCT, AM-A, JFM-R, AL-J, MAM-C, AL-C 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2096 66 585 SAT-496 PO03-02 Saturday 524 2012


525 ENDO12L_SAT-497 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) The Effect of Major Surgery on Glucocorticoid Concentrations Laura Jane Owen, Rachel Louise Jones, Timothy Strang, Lajos Szentgyorgyi, Gregory Cook, Maria Safar, Peter Trainer, Brian Keevil UHSM, Manchester, UK; UHSM, Manchester, UK; UHSM, Manchester, UK Introduction:[br]The stress response post-operatively increases glucocorticoid concentration and a higher cortisol response is associated with better recovery. The measurement of cortisol concentrations post-operatively can be unreliable due to altered binding protein concentrations. Free serum cortisol is felt to be the better marker of bio-active glucocorticoid concentrations but is difficult to measure. Salivary cortisol and cortisone have been shown to correlate well with serum free cortisol and is independent of binding protein concentrations. We investigated the use of salivary cortisol and cortisone in this group of patients.[br]Methods:[br]Patients undergoing cardiothoracic surgery without any history of glucocorticoid drug use or adrenal pathology were recruited into the study. Paired saliva and serum samples were collected pre- and post-operatively. Serum total cortisol, serum free cortisol, salivary cortisol and salivary cortisone were all measured by liquid chromatography tandem mass spectrometry. Saliva was collected using a disposable pipette after allowing the saliva to pool in the mouth for a few minutes.[br]Results:[br]A poor correlation was observed between salivary cortisol serum free cortisol (r = 0.34) and serum free cortisol and salivary cortisone (r = 0.28). A curvilinear correlation was seen between serum free cortisol and serum total cortisol as expected (r = 0.93). The range of pre-operative cortisol concentrations was 55-552 nmol/L. The range of post-operative cortisol concentrations was 35-1233 nmol/L. 70% of post-operative samples which were collected in the morning had cortisol concentrations [lt]500 nmol/L.[br]Discussion:[br]Despite previous reports promoting the use of salivary cortisol and cortisone as a marker of serum free cortisol a poor correlation was seen in this population. This may be due to the poor quality of saliva collected in this group and different methods for collecting saliva should be investigated. A significant proportion of patients had low cortisol concentrations post-operatively.[br][br]Disclosures: PT: Clinical Researcher, Ipsen, Versartis. Nothing to Disclose: LJO, RLJ, TS, LS, GC, MS, BK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 548 66 586 SAT-497 PO03-02 Saturday 525 2012


526 ENDO12L_SAT-498 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Characterization of the Novel Missense Mutation G250V in Type II 3[beta]-Hydroxysteroid Dehydrogenase (3[beta]-HSD2) Gene Found in a 46,XX (Female) Patient with Congenital Adrenal Hyperplasia (CAH) Maria Sonia Baquedano, Marta Ciaccio, Roxana Marino, Natalia Perez-Garrido, Pablo Ramirez, Marco A Rivarola, Alicia Belgorosky Hospital de Pediatria Garrahan, Buenos Aires, Argentina 3[beta]-HSD2 deficiency, a rare form of CAH, is characterized by varying degrees of salt loss and incomplete masculinization in males and mild virilization or normal external genitalia in females. 3[beta]-HSD2 gene mutations have been reported in a small number of affected females.[br]We report a 7-moth-old 46,XX girl who was referred because of premature pubarche and postnatal clitoromegaly. Hormonal profile showed inadequate production of glucocorticoids and very high serum ACTH levels (cortisol:4.8mcg/dl:ACTH: 2888pg/ml), increased serum 17OHProg(141ng/ml) and plasma renin levels (423.9ng/ml), in addition to very high levels of DHEAS (53000ng/ml).[br]Genetic analysis revealed a novel homozygous c.749G[rarr]T mutation in 3[beta]-HSD2 gene, resulting in a G250V change. This G is highly conserved in the vertebrate 3[beta]-HSD2 gene family and is located in the substrate-binding domain of the enzyme. As [italic]in silico[/italic] PolyPhen and SIFT analysis predicted G250V to be a damaging substitution, enzymatic activity was analyzed by in vitro analysis of mutant recombinant enzyme generated by site-directed mutagenesis after its transient expression in COS cells. Enzyme activity using 0.5 [micro]M pregnenolone as substrate in the medium after 6h revealed relative conversion rates of pregnenolone to progesterone of 78[plusmn]4% and 21[plusmn]1% by WT and G250V-3[beta]-HSD2 enzymes respectively. Using 0.5 [micro]M dehydroepiandrosterone as substrate, the relative conversion rate of dehydroepiandrosterone to androstenedione after 6h was 87[plusmn]8% and 23[plusmn]7% by WT and G250V-3[beta]-HSD2 enzymes, respectively. Immunofluorescence studies showed that both WT and mutant G250V-3[beta]-HSD2 protein colocalized with endoplasmic reticulum.[br]We identified a novel G250V 3[beta]-HSD2 gene mutation which causes an incomplete loss of enzymatic activity, explaining the compensated non-salt loss phenotype of CAH. Peripheral type 1, 3[beta]-HSD activity often complicates the hormonal diagnosis of this disorder. However, the discrepancy between the grossly elevated 17-OHP levels, as well as very high levels of DHEAS and the lack of ambiguous genitalia point against 21-hydroxylase deficiency. Flux via the adrenal [ldquo]backdoor[rdquo] pathway, which convert 17OHProg to dihydrotestosterone (DHT) has been recently implicated in human disorders of androgen excess. We hypothesized that this alternate pathway could not be activated in 3[beta]-HSD2 deficiency due to very low intraadrenal 17OHProg substrate levels, explaining mild virilization or even normal sexual differentiation in females.[br][br]Sources of Research Support: FONCYT,CONICET,Argentina.[br][br]Nothing to Disclose: MSB, MC, RM, NP-G, PR, MAR, AB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 331 66 587 SAT-498 PO03-02 Saturday 526 2012


527 ENDO12L_SAT-499 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Evaluation of Cortisol Resistance in Young Sedentary and Endurance-Trained Men Mahtab Niyyati, Ozan Y Suer, Qingxiang Wei, Lynnette K Nieman National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD OBJECTIVE: Some previous reports suggest central and peripheral glucocorticoid (GC) resistance and decreased negative cortisol feedback in athletes. We examined the hypothalamic-pituitary-adrenal axis of male exercisers (Es, running [ge]28 mi weekly) and nonexercisers (NEs, less than 1 h exercise weekly) with 0.25 mg overnight dexamethasone suppression test (DST) and baseline salivary cortisol. We evaluated glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) expression in PBMC and assessed their sensitivity to dexamethasone (D).[br]METHODS: In vitro sensitivity to GCs was assessed by measurement of D suppression of lipopolysaccaride (LPS) stimulated cytokines (IL-6, TNF-a, IFN-[gamma]) in PBMCs. Basal GR and MR expression and pre- and post-D target gene expression (glucocorticoid-induced leucine zipper (GILZ), FK506 binding protein 5 (FKBP5), IL-2) were assessed by RT-PCR. Salivary cortisol was measured at 2100 and 2300 on 2 separate days. Es ran on one day and not the other. Overnight 0.25 mg DST was performed. Unpaired student[apos]s t-test was used to compare the 2 groups.[br]RESULTS: 23 NEs and 17 Es participated. Age (E: 24.0 NEs: 25.0 yrs) and mean body mass index (Es: 23.3 NE: 24.2 kg/M2) were similar. Both groups demonstrated a decrease in PBMC cytokine responses with increasing D concentrations; the amplitude of cytokine release was similar in the two groups both at baseline and post D (P[gt]0.05). EC50 for D suppression of LPS-stimulated cytokines was similar (P[gt]0.05). There was no significant difference in the MR and GR expression between the two groups (P[gt]0.05). GILZ and FKBPS gene expression increased while IL2 decreased with higher D dosage in both groups; the EC50 was not significantly different. In Es nonrunning day 11 pm salivary cortisol was 118.8[plusmn]154.4 ng/dl (mean [underline]+ [/underline]SD)([lt]100.0 ng/dl), and in NEs day 1 salivary cortisol was 86.1[plusmn]49.2ng/dl (P [gt] 0.05). The running day salivary cortisol was 101.3[plusmn]148.9 compared to 88.0[plusmn]100.9 on NE day 2 (P [gt] 0.05). 19 NEs and 12 Es completed the DST. The post-DST cortisol levels were similar in the 2 groups (Es 9.9[plusmn]4.5vs NEs 9.7[plusmn]5.2 mcg/dl (5.0-25.0), mean[plusmn]SD, P=0.89). Cortisol and D levels were correlated significantly in NEs (r[sup]2[/sup] 0.5546, P=0.0137) but not in Es (r[sup]2[/sup] 0.1366, P=0.67).[br]CONCLUSION: Es and NEs had similar salivary cortisol, 0.25 mg DST, PBMC LPS secretion with D suppression, GR and MR receptor and target gene expression, suggesting similar sensitivity to endogenous glucocorticoid.[br][br]Sources of Research Support: The intramural program of NICHD, NIH.[br][br]Nothing to Disclose: MN, OYS, QW, LKN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1551 66 588 SAT-499 PO03-02 Saturday 527 2012


528 ENDO12L_SAT-500 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) BclI and N363S May Affect Cortisol Response after Dexamethasone Suppression Test and the Metabolic Profile in Patients with Adrenal Incidentaloma: Results from a Preliminary Study Giuseppe Reimondo, Daniela Giachino, Ilaria Micossi, Paola Berchialla, Simona Varacalli, Barbara Allasino, Anna Pia, Federica Laino, Giulia Peraga, Paolo Cosio, Chiara Sciolla, Mario De Marchi, Massimo Terzolo Medicina Interna 1 a Indirizzo Endocrinologico - AOU San Luigi, Orbassano (TO), Italy; Genetica Medica - AOU San Luigi, Orbassano (TO), Italy; Statistica - AOU San Luigi, Orbassano (TO), Italy The effects of glucocorticoids are mediated by the glucocorticoid receptors (GR) and some single nucleotide polymorphisms (SNPs) have been associated to enhanced sensitivity (N363S, BclI) or reduced sensitivity (ER22/23EK) to glucocorticoids.[br]Aims of the present study were: 1) to assess the frequency of N363S, BclI and ER22/23EK in 46 patients (37 F and 9 M, median age 44 years) with Cushing[apos]s syndrome (CS) and 73 patients (42 F and 31 M, median age 61 years) with adrenal incidentaloma (AI) compared to 186 healthy subjects; 2) to evaluate a possible correlation between the different SNPs and patient phenotype.[br]In all patients we evaluated clinical and demographic parameters, urinary free cortisol (UFC) and cortisol after 1 mg dexamethasone suppression test (DST). DNA was extracted from peripheral blood leukocytes using PCR. The PCR product was digested with 1 U of TasI at 65[deg]C overnight, and the accuracy of genotyping was confirmed by sequencing analysis.[br]We did not observe any differences between patients and controls for the frequency of N363S (CS 0%, AI 7.6%, controls 9.7%), BclI (CS 58.6%, AI 55.7%, controls 55.4%) and ER22/23EK (CS 5.2%, AI 1.3%, controls 3.8%). Although not statistically different, we observed a reduced frequency of BclI in patients with AI and cortisol [gt] 5 mcg/dl after DST (5.1% vs 10.3% in the remainders). Moreover, 4 of the 5 patients with AI carrying the N363S SNP had an overt metabolic syndrome despite normal UFC levels and cortisol suppressibility after DST. The low frequency of the ER22/23EK SNP precluded a meaningful statistical analysis.[br]Our data suggest that the evaluated SNPs are not involved in the pathogenesis of CS or AI. In patients with CS, we did not observe any genotype/phenotype correlation, since the overt hypercortisolism likely overruns any modulatory effect of the SNPs on clinical parameters. In AI patients, who present only a mild hypercortisolism, if any, BclI may affect cortisol suppression after DST, while N363S may be associated to a worsen metabolic profile.[br][br]Nothing to Disclose: GR, DG, IM, PB, SV, BA, AP, FL, GP, PC, CS, MDM, MT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1204 66 589 SAT-500 PO03-02 Saturday 528 2012


529 ENDO12L_SAT-501 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) The Role of Basal Serum Cortisol in the Assessment of Pituitary-Adrenal Insufficiency Fatima Imran, Mohammad Alotaibi, Sam Stewart, Stephanie M Kaiser, David B Clarke, Thomas P Ransom, Syed Ali Imran Dalhousie University, Halifax, Canada; Dalhousie University, Halifax, Canada; Dalhousie University, Halifax, Canada The role of morning (basal) serum cortisol as a marker of HPA (hypothalamic pituitary adrenal) function has been reported previously but those studies were conducted mostly in the context of pituitary surgery where insulin tolerance tests (ITT) were used as the gold standards. The cut-off values for basal cortisol excluding the need for a dynamic test in those studies ranged between 100 - 138 nmol/L (lower) and between 300 - 495 nmol/L (upper). In our centre, we have been using lower and upper cut-off values of [lt]100 nmol/L and [gt]300 nmol/L, respectively, based on the original data from Hagg et al (1). For dynamic testing, we use the standard 250 mcg ACTH test, which is an effective alternative to the more cumbersome ITT (2). For this study, we analyzed consecutive patients since 2008 who had been assessed through our neuropituitary clinic for a provisional diagnosis of adrenal insufficiency. Those patients, who based on the above criteria had undergone basal (0900) serum cortisol test followed by an ACTH test were identified and their data were analyzed. A total of 73 patients (46 females) underwent both tests; the primary diagnoses were pituitary tumour (34), fatigue (15), radiation therapy (7), iatrogenic adrenal insufficiency (7), hypotension and vomiting (3), non-pituitary sellar mass (4) and hypopituitarism (3). Post-ACTH cortisol peaks of either [gt]500 or [gt]550 nmol/L have previously been reported as standards to exclude HPA insufficiency. Based on post ACTH peaks of [gt]500 nmol/L and [gt]550 nmol/L, the lower cut-off values for 0900 basal cortisol providing 100% sensitivity were 121 and 128 nmol/L, respectively, while the upper cut-off providing 100% specificity was 243 nmol/L for both. Optimal values of serum cortisol providing the best fit were calculated through the ROC analysis. A 0900 serum cortisol value of 168 nmol/L while using a post-ACTH peak of [gt]500 nmol/L provided a sensitivity of 86% and specificity of 59% whereas a 0900 cortisol of 145 nmol/L while using a post-ACTH peak of [gt]550 nmol/L provided a sensitivity of 98% and a specificity of 40%. In conclusion, these results suggest that 0900 serum cortisol levels of [lt]128 and [gt]243 nmol/L can be used to effectively identify those who require dynamic testing. The upper cut-off 0900 cortisol value from our data is significantly lower than the presently suggested values, indicating that many patients may currently be unnecessarily undergoing ACTH testing.[br][br](1) Hagg E et al., Clin Endocrinol. 1987: 221. (2) Abdu TA et al., J Clin Endocrinol Metab. 1999: 838.[br][br]Nothing to Disclose: FI, MA, SS, SMK, DBC, TPR, SAI 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 134 66 590 SAT-501 PO03-02 Saturday 529 2012


530 ENDO12L_SAT-502 POSTER SESSION: Control of the Hypothalamo-Pituitary-Adrenal Axis (1:30 PM-3:30 PM) Effects of Growth Hormone Excess on the Synthesis and the Metabolism of Cortisol Tomoatsu Mune, Tetsuya Suwa, Shin-Ichi Kawachi, Jun Takeda, Takatoshi Anno, Shinji Kamei, Mitsuru Hashiramoto, Michihiro Matsuki, Kohei Kaku Kawasaki Medical School, Kurashiki, Japan; Gifu University, Gifu, Japan The growth hormone (GH) - insulin-like growth factor 1 axis plays an important role in modulating the peripheral metabolism of glucocorticoids mainly through its effect on the type 1 isozyme of 11[beta]-hydroxysteroid dehydrogenase (11[beta]HSD), which act as a reductase catalyzing the conversion of inactive cortisone (E) to active cortisol (F). Here we analyzed the effects of GH excess on the pituitary-adrenal axis as well as F-E status. We measured serum 17-hydroxyprogesterone (17OHP4), 11-deoxycortisol (S), F and E by the method using liquid chromatography-tandem mass spectrometry (LC/MS/MS) in 15 (male/female 4/11, age 30-73yr) patients with acromegaly before and after treatment. The ratio of S/17OHP4, F/S and F/E was calculated as a marker of 21-hydroxylase, 11[beta]-hydroxylase (the final step of F production) and 11[beta]HSD type 1 activity, respectively.[br]Serum levels of each steroid (mean[plusmn]SD in ng/ml) were as follows; 17OHP4 0.39[plusmn]0.32/0.41[plusmn]0.39 (before/after), S 0.23[plusmn]0.28/0.20[plusmn]0.22, F 94.9[plusmn]44.7/90.7[plusmn]33.3, E 27.7[plusmn]6.3/23.5[plusmn]5.4, and the decrease in E after cure of the disease was significant (P=.023). There were no differences in the S/17OHP4 ratio or the F/S ratio, but the F/E ratio was increased from 3.33[plusmn]1.24 to 3.83[plusmn]0.97(P=.047)after treatment. Plasma ACTH did not change significantly, but the ratio of plasma ACTH to F was decreased from 4.3[plusmn]2.1 to 3.3[plusmn]1.5 (P=.041).[br]At the active phase (before treatment), serum GH levels were positively correlated with levels of plasma ACTH (r[sup]2[/sup] =.26) and serum S (r[sup]2[/sup] =.36). Serum GH levels was positively correlated with the S/17OHP4 (r[sup]2[/sup] =.52) and F/E ratio (r[sup]2[/sup] =.39) but negatively correlated with the F/S ratio (r[sup]2[/sup] =.20).[br]These data suggest that excess GH inhibits the reductase activity of 11[beta]HSD type 1 concomitantly with enhanced ACTH drive probably at the central site (hypothalamus or pituitary). However, the positive correlation between serum GH and the F/E ratio before treatment rather suggests the stimulation of the reductase activity. Influence of ACTH drive, enhanced 21-hydroxylation with 11[beta]-hydroxylase inhibition by GH axis should also be considered. Further study will be necessary.[br][br]Nothing to Disclose: TM, TS, S-IK, JT, TA, SK, MH, MM, KK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1345 66 591 SAT-502 PO03-02 Saturday 530 2012


531 ENDO12L_SAT-503 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Novel [italic]KAL1[/italic] Sequence Variants Associated with Septo-Optic Dysplasia (SOD) in Three Female Patients Mark James McCabe, Youli Hu, Louise Cheryl Gregory, Ajay Thankamony, Ieuan Hughes, Sharron Townshend, Pierre-Marc Bouloux, Mehul T Dattani UCL-Institute of Child Health, London, UK; Royal Free Hospital and University College Medical School, University College London, London, UK; University of Cambridge, Addenbrookes Hospital, Cambridge, UK; Princess Margaret Hospital for Children, Subiaco, Australia [bold]Background and aims: [/bold][italic]KAL1[/italic] is essential for GnRH neuronal migration and olfactory bulb development with mutations accounting for 5% of patients with Kallmann syndrome (KS), characterized by combined hypogonadotrophic hypogonadism and anosmia. [italic]KAL1 [/italic]is located in the X-chromosome pseudoautosomal region which may account for the recent report of female KS patients exhibiting [italic]KAL1 [/italic]variations.[sup]1[/sup][br]Increasing evidence of overlapping genotypes between KS and hypopituitarism including septo-optic dysplasia (SOD) and craniofacial defects is consistent with the known overlap in phenotypes. Therefore, we aimed to screen 421 patients with the latter (M:F 1.1:1) for mutations in [italic]KAL1[/italic].[br][bold]Methods: [/bold]We used direct sequencing analysis to screen the coding region of [italic]KAL1[/italic] and optimized [italic]in vitro[/italic] assays to analyze the functional consequences of any variants identified. These included qualitative investigations of mutational effects on protein secretion by immunocytochemistry (ICC) and western blot using GFP-labelled KAL1 in Cos7 cells, in addition to a quantitative luciferase-reporter assay in L6-myoblasts overexpressing FGFR1 [italic]in vitro[/italic].[br][bold]Results: [/bold]Three female patients with SOD tested positive for sequence variants in [italic]KAL1[/italic], absent in 480 controls, at highly conserved residues; p.K185N (n=1) and p.P291T [novel; n=2 (sisters)]. All three children had optic nerve hypoplasia and GH deficiency, with the p.K185N variant also being associated with TSH deficiency and an ectopic posterior pituitary. The p.P291T variant is located between the first two fibronectin domains; we observed a qualitative decrease in secretion of the mutant protein as shown by its retention in Cos7 cells and a 40% decrease in transcriptional activity (p[lt]0.001). Secretion of p.K185N was unaffected but the variant was associated with a 21% decrease in transcriptional activity (p[lt]0.01). This variant is located between the WAP and first fibronectin domains of KAL1 and is predicted to affect the binding (studies ongoing) of the protein to FGFR1 and heparan sulfate. Both variants were inherited from the unaffected mothers. The presence of the variant may reflect variable penetrance or digenicity/oligogenicity in the affected individuals, none of whom exhibit variations in any of the known KS genes.[br][bold]Conclusion:[/bold] We implicate [italic]KAL1 [/italic]in females with hypopituitarism/SOD for the first time to our knowledge, reflecting an overlap between KS and SOD that has also been observed with [italic]FGF8[/italic], [italic]FGFR1[/italic] and [italic]PROKR2[/italic] variants.[br][br](1) Shaw et al., J Clin Endocrinol Metab 2011; 96:E566.[br][br]Nothing to Disclose: MJM, YH, LCG, AT, IH, ST, P-MB, MTD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1533 67 592 SAT-503 PO20-02 Saturday 531 2012


532 ENDO12L_SAT-504 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Neural Pathways by Which Leptin Modulates GnRH Neurons in the Sheep: Direct and Indirect Activation of AgRP, POMC and Kisspeptin (KNDy) Neurons Maria Cernea, Lique M Coolen, Rebecca Phillips, Michael N Lehman University of Michigan, Ann Arbor, MI; University of Michigan, Ann Arbor, MI Leptin is an important metabolic signal of energy stores and an indicator to the reproductive axis whether sufficient stores are available to permit reproduction. Leptin modulates pulsatile gonadotropin secretion from the anterior pituitary via the regulation of hypothalamic gonadotropin releasing hormone (GnRH) neurons[sup]1[/sup]. The arcuate (ARC) KNDy neurons (co-expressing the neuropeptides [underline]k[/underline]isspeptin, [underline]n[/underline]eurokinin B/[underline]dy[/underline]norphin) are important afferents involved in the control of pulsatile GnRH secretion[sup]2[/sup] and may comprise one of the pathways by which leptin influences reproduction. Neuronal activation by leptin can be studied by examining phosphorylation of STAT3 (pSTAT3), a direct consequence of the binding of leptin to its receptor. In addition, induction of the immediate early gene product, cFos can be used as an indicator of general activation. We examined leptin-induced pSTAT and cFos in sheep brain to delineate the neural pathways by which leptin might regulate GnRH secretion in this species. Adult, ovary-intact ewes received injections of either leptin (1mg/kg, i.v.) or vehicle, and two hours later were euthanized and their brains perfused for immunocytochemistry. Alternate series of sections were processed for dual immunoperoxidase detection of pSTAT or cFos with either kisspeptin, agouti-related peptide (AgRP), proopiomelanocortin (POMC), or GnRH. Leptin-treated sheep showed pSTAT expression in a majority of AgRP (84%) and POMC (30%) neurons, but not in kisspeptin or GnRH neurons; control, vehicle-injected animals exhibited no pSTAT in any of these neurons. By contrast, leptin induced cFos expression in a majority of ARC kisspeptin (70%) and POMC (92%) neurons, while control animals showed 20-30% basal levels of co-expression. AgRP neurons showed high levels expression of cFos in both leptin and control animals (70-90%). GnRH neurons were not examined for cFos expression. In summary, leptin appears to directly activate AgRP and POMC, but not kisspeptin and GnRH, neurons by activation of leptin receptors and STAT3. Moreover, ARC kisspeptin neurons are also activated by leptin but indirectly, presumably by afferents from other cells that in turn are leptin-responsive. These results suggest that leptin may modulate GnRH secretion in sheep by a pathway that starts with AgRP and POMC neurons, but may also include projections to KNDy cells and their inputs to GnRH neurons.[br][br]1. Quennell, J.H., Mulligan, A.C., Tups, A., Liu, X., Phipps, S.J., Kemp, C.J., Herbison, A.E., Grattan, D.R., and Anderson, G.M., 2009. Leptin indirectly regulates gonadotropin-releasing hormone neuronal function. Endocrinology; 150: 2805[ndash]2812. 2. Lehman, M.N., Coolen, L.M., and Goodman, R.L., 2010. Minireview: kisspeptin/neurokinin B/dynorphin (KNDy) cells of the arcuate nucleus: a central node in the control of gonadotropin-releasing hormone secretion. Endocrinolog; 151(8): 3479-89.[br][br]Sources of Research Support: P01 HD044232 (Project II) to M.N.L.[br][br]Nothing to Disclose: MC, LMC, RP, MNL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2155 67 593 SAT-504 PO20-02 Saturday 532 2012


533 ENDO12L_SAT-505 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Colocalization of Met-Enkephalin, but Not Galanin or Tyrosine Hydroxylase, within Kisspeptin Neurons in the Sheep Christina M Merkley, Lique M Coolen, Robert L Goodman, Michael N Lehman University of Michigan, Ann Arbor, MI; University of Michigan, Ann Arbor, MI; West Virginia University, Morgantown, WV Kisspeptin neurons are key regulators of reproduction, playing a role in both surge and pulsatile secretion of GnRH (1). The colocalization of neurokinin B and dynorphin is unique to arcuate nucleus (ARC) kisspeptin neurons (the KNDy neurons) and is not seen in kisspeptin cells in the preoptic region (POA) (2). To gain a better understanding of the physiological role of each population in the sheep, we examined possible colocalization of other neuropeptides/transmitter enzymes recently shown to be coexpressed in rodent kisspeptin neurons (3): met-enkephalin (met-ENK), galanin (GAL), and tyrosine hydroxylase (TH; marker of dopaminergic neurons). Brain sections from OVX+E ewes perfused during the breeding season (n=4) were processed for dual immunofluorescent detection of kisspeptin with either met-ENK, GAL or TH. Sections from the ARC (rostral, middle, and caudal divisions) and the POA were examined by confocal microscopy, and the percentage of kisspeptin cells co-expressing each neuropeptide/transmitter enzyme was calculated. Only a very small percentage ([lt]3%) of kisspeptin cells in either the POA or ARC colocalized GAL despite the presence of numerous single-labeled GAL-positive cell bodies in both regions. In addition, we found no instances of dual labeled kisspeptin/TH neurons within the POA or ARC, although single-labeled TH-positive neurons were found in both regions (A14 dopamine cells in the POA; A12 cells in the ARC). By contrast, 20% and 8-12% of POA and ARC kisspeptin neurons, respectively, colocalized met-ENK; single-labeled met-ENK neurons were also seen in the same regions, and their percentage of colocalization with kisspeptin was similar (18% and 15-25% for POA and ARC met-ENK neurons, respectively). In summary, we report that a small but consistent subset of ARC KNDy and POA kisspeptin neurons co-express met-ENK, and not galanin and TH. Since endogenous opioid peptides play an important role in steroid feedback control of GnRH, co-release of met-ENK from a subset of kisspeptin neurons may serve to modulate the influence of kisspeptin and other co-localized neuropeptides (NKB, dynorphin) on GnRH secretion. It remains to be explored whether expression of met-ENK, like that of kisspeptin, is regulated by gonadal steroid hormones in neurons of the sheep POA and ARC.[br][br]1. Lehman, Merkley et al., Brain Research 2010; 1364:90-102. 2. Goodman et al., Endocrinology 2007; 148(12):5752-5760. 3. Porteous et al., J Comp Neurol 2011; 519(17):3456-69.[br][br]Sources of Research Support: NIH R01HD33916 to M.N.L. and R.L.G.[br][br]Nothing to Disclose: CMM, LMC, RLG, MNL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2156 67 594 SAT-505 PO20-02 Saturday 533 2012


534 ENDO12L_SAT-506 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) A Unique Population of Prepro-Thyrotropin-Releasing Hormone[ndash]Expressing Neurons in the Lateral Hypothalamus That Are Activated by Leptin and Altered by Prenatal Glucocorticoid Exposure David Louis Carbone, Robert James Handa University of Arizona College of Medicine-Phoenix, Phoenix, AZ The neuropeptide thyrotropin-releasing hormone (TRH) plays an important role in controlling energy balance by regulating the hypothalamic-pituitary-thyroid axis, as well as through direct effects on the CNS. In order to better understand the effects of TRH on CNS control of energy balance, we used immunocytochemistry (ICC) to map the distribution of neurons expressing the TRH precursor peptide, prepro TRH (ppTRH) in the paraventricular nucleus and surrounding regions of the male Sprague Dawley rat hypothalamus. This study identified a unique population of ppTRH-immunoreactive (ir) neurons and fibers in the juxtaparaventricular region of the lateral hypothalamus (LHAjp). We also observed a decrease in ppTRH-ir neurons and fibers in the LHAjp in adult male rats which had been exposed during fetal development to excess glucocorticoid (GC). Our GC exposure paradigm consisted of subcutaneous injections of the synthetic GC dexamethasone (DEX; 0.4mg/kg BW) to pregnant dams between gestational days 18-21, and resulted in increased feeding activity in the adult offspring. Based on these observations, we tested the hypothesis that the ppTRH-ir neurons within the LHAjp are a component of neuronal circuitry involved in the regulation of feeding activity by administering leptin (2.5 [mu]g/kg BW) to adult male Sprague Dawley rats, which were then killed two hours later. Using ICC, we observed that leptin administration resulted in the co-expression of c-Fos by ppTRH-ir neurons in the LHAjp, indicating that these cells are targets for the satiety signal leptin. Because existing reports describe the presence of neuronal connections between the LHAjp and the arcuate nucleus of the hypothalamus (ArcN), which plays a crucial role in regulating feeding activity, we measured the expression of TRH receptors (Trhr1, Trhr2) in the ArcN using RT-qPCR. This revealed the expression of mRNAs for both receptors. Taken together, these data suggest a role for the LHAjp in regulating feeding activity perhaps through cross-talk with the ArcN. Although the biological role for the LHAjp remains poorly understood, our data suggest that this region of the brain may influence energy homeostasis through neuroendocrine modulation of specific nuclei throughout the CNS.[br][br]Sources of Research Support: NIH NS039951 and MH082679.[br][br]Nothing to Disclose: DLC, RJH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1770 67 595 SAT-506 PO20-02 Saturday 534 2012


535 ENDO12L_SAT-507 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Kp-16 and Kp-13 Are the Most Abundant Kisspeptin Isoforms Present in Sheep Hypothalamus Alain Caraty, Jerome Leprince, Didier Lomet, Benjamin Lefranc, Adele Bourmaud, Marie-Emilie Sebert, Massimiliano Beltramo, Hubert Vaudry INRA, Nouzilly, France; University of Rouen, Mont-Saint-Aignan, France Various forms of Kisspeptin (Kp) have been purified from the human placenta: KP-54, KP-14, KP-13 and KP-10. They derive from the proteolytic processing of a 145-amino acid (aa) precursor protein encoded by the KISS1 gene. However, whether similar cleavage of the precursor occurs in the hypothalamus remains to be determined. To this aim, hypothalamus from ewes were collected either during the breeding season (n =110, A) or the anestrous season (n =110, B). Hypothalami were pooled, homogenized in boiling acetic acid and extracted on C18 Sep-Pack cartridges. Lipids were removed by dichloromethane extraction and extracts were dried. Dried extracts were dissolved in 0.1% TFA/water, injected on a C18 reversed-phase HPLC column (Vydac 218TP54, 0.46 x 25 cm), eluted with a 20-30% acetonitrile gradient and fractions were collected every 20 sec. Kp immunoreactive (IR) material was measured in these fractions using a Kp RIA with an antibody directed against the common C-terminal part of the molecules. Five major IR peaks, with similar retention time (RT) and similar amplitude, were observed for extracts A and B. In 4 out of 5 peaks, RTs matched those of the synthetic ovine Kp fragments: oKp-10, oKp-13, oKp-14 and oKp-16. To further characterize the samples, fractions corresponding to each peak from the two extracts were pooled and submitted to mass spectrometry analysis (Maldi, Trap and Orbitrap). Identity of the 4 short ovine isoforms, previously determined according to their RTs, was definitively confirmed by the mass. The 5th unidentified peak had a molecular weight (5620 dalton) identical to that of the 53-aa peptide corresponding to the predicted sequence of the sheep Kp-53. The proportion of each peak was estimated according to the cross-reactivity with the antibody and the molecular weight. In contrast to what was reported in human placenta, the longest isoform (54 aa in human and 53 aa in ewe) was not the major Kp isoform present in the sheep hypothalamus (15%); oKp-16, oKp-14, oKp-13 and oKp10 being respectively 42, 8, 33 and 2% of all Kp molecules. These data identify for the first time a 53-aa peptide as the longest Kp molecule in the sheep hypothalamus. They also indicate that the Kp precursor is mainly processed into low molecular weight peptides mainly oKp-16 and oKp-13. Further experiments are now required to determine if these peptides have distinct physiological role in controlling brain function.[br][br]Sources of Research Support: ANR FrenchKiss.[br][br]Nothing to Disclose: AC, JL, DL, BL, AB, M-ES, MB, HV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 493 67 596 SAT-507 PO20-02 Saturday 535 2012


536 ENDO12L_SAT-508 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Different Effects of Electrical and Manual Acupuncture Stimulation on Estrous Cyclicity and Neuroendocrine Function in Rats with DHT-Induced Polycystic Ovary Syndrome Yi Feng, Julia Johansson, Ruijin Shao, Louise Manneras Holm, Hakan Billig, Elisabet Stener-Victorin Institute of Neuroscience and Physiology, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden; Institutes of Brain Science, Shanghai, China; First Affiliated Hospital, Heilongjiang University of Chinese Medicine, Harbin, China Both low-frequency electro-acupuncture (EA) and manual acupuncture improve menstrual frequency and decrease circulating androgens in women with polycystic ovary syndrome (PCOS). We sought to determine whether low-frequency EA is more effective than manual stimulation in regulating disturbed estrous cyclicity in rats with PCOS induced by 5[alpha]-dihydrotestosterone (DHT). To identify the central mechanisms of the effects of stimulation, we assessed hypothalamic mRNA expression of molecules that regulate reproductive and neuroendocrine function. From age 70 days, rats received 2-Hz EA or manual stimulation of the needles five times/week for 4[ndash]5 weeks; untreated rats served as controls. Specific hypothalamic nuclei were obtained by laser microdissection, and mRNA expression was measured with TaqMan low-density arrays. Untreated rats were acyclic. During the last 2 weeks of treatment, seven of eight (88%) rats in the EA group had epithelial keratinocytes, demonstrating estrous cycle change (p = 0.034 vs. controls). In the manual group, five of nine (56%) rats had estrous cycle changes (ns vs. controls). mRNA expression of the opioid receptors Oprk1 and Oprm1 in the hypothalamic arcuate nucleus was lower in the EA group than in untreated controls. mRNA expression of the steroid hormone receptors Esr2, Pgr, and Kiss1r was lower in the manual group than in the controls. In rats with DHT-induced PCOS, low-frequency EA restored disturbed estrous cyclicity but did not differ from manual stimulation group. Thus EA cannot be considered superior to manual stimulation. The effects of low-frequency EA may be mediated by central opioid receptors, while manual stimulation may involve regulation of steroid hormone/peptide receptors.[br][br]Sources of Research Support: This study was supported by grants from the Swedish Medical Research Council (Project No. 2008-72VP-15445-01A); Novo Nordisk Foundation; Wilhelm and Martina Lundgrens[apos]s Science Fund; Hjalmar Svensson Foundation; Adlerbert Research Foundation; Swedish federal government under the LUA/ALF agreement ALFFGBG-10984 and 136481 (E. S-V), Chinese Special Fund for Postdoc (No. 200801170) and National Natural Science Foundation of China) (No. 81001544/H2718) (Y. F.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.[br][br]Nothing to Disclose: YF, JJ, RS, LMH, HB, ES-V 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1368 67 597 SAT-508 PO20-02 Saturday 536 2012


537 ENDO12L_SAT-509 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) The Direct Effects of Palmitate and Omega-3 Fatty Acids on the Molecular Circadian Clock in Immortalized, Clonal Hypothalamic Neurons James Greco, Leigh Wellhauser, Denise D Belsham University of Toronto, Toronto, Canada; Toronto General Hospital Research Institute, University Health Network, Toronto, Canada With obesity, excessive nutrient levels can lead to metabolic disturbances and the progression of pathologies, such as diabetes and heart disease. The obese state has also been shown to have a reciprocal relationship with circadian rhythms. Recent studies have demonstrated that atypical circadian rhythms result in obese phenotypes and similarly, metabolic dysregulation negatively shifts the circadian clock. To date, the molecular mechanisms underlying these phenomena are not well understood and are difficult to study in vivo. In order to address this issue, our laboratory has generated an array of immortalized, clonal, neuronal, hypothalamic cell lines through retroviral transfection. Using real time RT-PCR, we have identified that the mHypoE 37 cell line expresses a number of neuropeptides linked to obesity such as neuropeptide Y (NPY), agouti-related peptide (AgRP), urocortin2, ghrelin, as well as important receptors, GPR210, insulin receptor, leptin long-form receptor, hypocretin receptor 1 and 2. It also expresses key circadian molecular components, Bmal1, Per2, and Rev-erb[alpha], all of which oscillate with an approximate 24 hour period. Previously, we have demonstrated that the saturated fatty acid, palmitate (200 [mu]M) attenuates insulin signalling, generates ER stress, induces apoptosis, and increases the orexigenic neuropeptide, NPY. Further, palmitate disrupts cellular circadian gene expression. Conversely, unsaturated omega-3 fatty acids have been shown to have protective effects against the development of insulin resistance. Preliminary studies indicate that administration of 12 [mu]M of omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), can significantly suppress transcript levels of the orexigenic neuropeptide AgRP. Provided the opposing effects induced by saturated versus unsaturated fatty acids, we hypothosize that palmitate will hinder the circadian molecular clock in the mHypoE 37 cell model, whereas co-treatment with omega-3 fatty acids will have restorative effects. Furthermore, due to the variance in neuronal sensing of saturated and unsaturated fatty acids, we are delineating the differential signalling cascades by which each type of fatty acid alters circadian transcript levels. Our novel hypothalamic cell models serve as optimal models to study the effects of nutrient excess on the cellular circadian clock, thereby further defining the complex interplay between circadian rhythms and metabolic status.[br][br]Sources of Research Support: CIHR, NSERC, CFI, and the CRC Program.[br][br]Nothing to Disclose: JG, LW, DDB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2105 67 598 SAT-509 PO20-02 Saturday 537 2012


538 ENDO12L_SAT-510 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Estradiol Inhibits AMPK and LKB1 in Cultured Hypothalamic Cells Jessica E Healy, Robert J Handa University of Arizona College of Medicine-Phoenix, Phoenix, AZ Obesity is of growing concern in Western society, mainly due to its associated pathologies, which include diabetes, heart disease, and atherosclerosis. During menopause in women, a decrease in circulating estradiol may lead to hyperphagia and obesity, as estradiol is anorexigenic in rodents and humans. At least two neuropeptides involved in the regulation of food intake (NPY and POMC) appear to be estrogen regulated, and are also regulated by the cellular energy sensing enzyme AMP-activated protein kinase (AMPK). AMPK is phosphorylated (pAMPK) by the enzyme liver kinase B1 (LKB1), and activated when a decrease in the AMP:ATP ratio signals low endogenous energy.[br]In these studies, we examined the possibility that estradiol exercises its anorexigenic effect through inhibition of AMPK activity resulting in decreases in food intake, as ovariectomized female rats (lacking estradiol) become obese and have increased expression of hypothalamic pAMPK. Treatment of the N38 hypothalamic cell line with estradiol (10nM) resulted in a decrease in the proportion of active to inactive AMPK, and decreases in the relative abundance and mRNA of NPY, in effect signaling an environment of high available energy. This response occurred within 10 minutes and persisted for up to 48 hours. Consequently, we hypothesized that estradiol decreases expression of LKB1, thereby decreasing phosphorylation and activation of AMPK. Treatment of N38 cells with estradiol (10nM) caused a decrease in LKB1 within 10 minutes, consistent with its effects on AMPK. These data point to a potential mechanism by which estradiol can impact energy balance: by preventing the phosphorylation and activation of AMPK. A better understanding of the cellular pathways underlying food intake may assist in the development of possible pharmacological interventions for the pathologies associated with the condition of obesity.[br][br]Sources of Research Support: NIH Grant NS039951 awarded to RJH.[br][br]Nothing to Disclose: JEH, RJH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2171 67 599 SAT-510 PO20-02 Saturday 538 2012


539 ENDO12L_SAT-511 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Developmental Programming Affects Female Pubertal Onset and Estrous Cyclicity Syann Lee, Carol F Elias, Joel K Elmquist University of Texas Southwestern Medical Center, Dallas, TX Studies have suggested that maternal diet during early development, known as developmental programming, can have long term effects on the offspring[apos]s metabolism with increased sensitivity to diet induced obesity. Here we show that female mice, born from mothers fed a high fat diet, have increased body weight and elevated levels of leptin at weaning. Additionally, these mice have advanced vaginal opening and abnormal estrous cycles. F2 generation mice, descended from programmed F1 females, maintain an increased sensitivity to diet induced obesity, but have normal reproductive parameters. These results suggest that the neonatal environment plays an important role in establishing female pubertal onset and estrous cyclicity.[br][br]Nothing to Disclose: SL, CFE, JKE 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2169 67 600 SAT-511 PO20-02 Saturday 539 2012


540 ENDO12L_SAT-512 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Pineal Hypoplasia and Decreased Melatonin in Patients with [italic]PAX6[/italic] Haploinsufficiency Due to WAGR Syndrome or [italic]PAX6[/italic] Mutations Alyson E Hanish, John A Butman, Amanda E Huey, Mark D Lee, Emily Yin, Lindsay A Hunter, Melanie D Hicks, Tanvee Singh, Matthew Tsang, Joan C Han Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD; National Institute of Nursing Research, National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD Background: WAGR syndrome (Wilms tumor, aniridia, genitourinary anomalies, mental retardation) is caused by heterozygous 11p13 deletions involving [italic]WT1[/italic] and [italic]PAX6[/italic]. Isolated aniridia is frequently caused by [italic]PAX6[/italic] haploinsufficiency (+/-) due to point mutations within [italic]PAX6[/italic]. Pineal gland hypoplasia has been previously reported [1] in a subpopulation of [italic]PAX6[/italic]+/- subjects, but melatonin has not been studied in humans with [italic]PAX6[/italic] mutations. The pineal gland is the primary source of melatonin, a hormone involved in circadian regulation. We hypothesized that melatonin and its urinary metabolite, 6-sulfatoxymelatonin (6SM), will be lower and that sleep dysfunction would be higher in WAGR/[italic]PAX6[/italic]+/- subjects as compared to healthy controls.[br]Methods: Groups of 39 WAGR subjects (age 14.0[plusmn]8.6y), 13 [italic]PAX6[/italic]+/- subjects (28.9[plusmn]16.2y), and 20 healthy controls (15.1[plusmn]7.3y) were evaluated during an inpatient admission. Pineal gland appearance was evaluated by brain MRI. Serum melatonin was drawn at 8AM and midnight (dark room from an IV). Urine was collected at 1st AM void and during the evening. Serum melatonin and 6SM were measured by ELISA. The Child Sleep Habits Questionnaire (CSHQ) [2] was administered for subjects [lt]18y. ANCOVAs compared groups (covariates: age, sex, [amp] BMI-Z).[br]Results: The prevalence of pineal gland hypoplasia/non-visualization was higher in WAGR [58% (95%CI: 39-75), p[lt]0.001] and [italic]PAX6[/italic]+/- [45% (95%CI: 17-77), p=0.005] vs. controls [0% (95%CI: 0-20)]. Midnight melatonin was lower in WAGR [adjusted mean[plusmn]SEM: 48[plusmn]14pg/mL, p=0.003] and [italic]PAX6[/italic]+/- [52[plusmn]16pg/mL: p=0.008] vs. controls [113[plusmn]13pg/mL]. Midnight melatonin was [sim]2-fold lower in those with vs. without pineal hypoplasia (p=0.02). The 8AM melatonin values were similar among groups (p=0.65), and [sim]2-fold lower than midnight values (p=0.01). Morning urinary 6SM levels (reflecting nighttime production) were lower in WAGR [18.3[plusmn]9ng/mgCr, p=0.006)] and [italic]PAX6[/italic]+/- [10.5[plusmn]9ng/mgCr, p=0.003] as compared to controls [63.9[plusmn]9ng/mgCr]. Evening 6SM levels were similar among groups (p=0.20), and [sim]3-fold lower vs. AM void (p=0.02). CSHQ score was 48.5[plusmn]1.7 for WAGR/[italic]PAX6[/italic]+/- vs. 42.6[plusmn] 2.7 in controls, but was not statistically different (p=0.08).[br]Conclusions: Pineal hypoplasia occurred in 55% of WAGR/[italic]PAX6[/italic]+/- subjects, and was associated with lower melatonin production. In WAGR/[italic]PAX6[/italic]+/- children, there was a trend toward greater sleep disturbance. Our findings support the view that [italic]PAX6[/italic] plays an important role in pineal development and function.[br][br][1] Mitchell, T. N., Free, S. L., Williamson, K. A., Stevens, J. M., Churchill, A. J., Hanson, I. M., Sisodiya, S. M. (2003). Polymicrogyria and absence of pineal gland due to PAX6 mutation. Ann Neurol, 53(5), 658-663. [2] Owens J.A., Spirito A., [amp] McGuinn M. The Children[apos]s Sleep Habits Questionnaire (CSHQ): psychometric properties of a survey instrument for school-aged children. Sleep 2000;23:1043-51.[br][br]Sources of Research Support: This study was supported by the Intramural Research Program of the National Institute of Child Health and Human Development and National Institute of Nursing Research, NIH.[br][br]Nothing to Disclose: AEH, JAB, AEH, MDL, EY, LAH, MDH, TS, MT, JCH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1801 67 601 SAT-512 PO20-02 Saturday 540 2012


541 ENDO12L_SAT-513 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Obestatin Promotes Adult Rat Hippocampal Progenitor Cell Proliferation and Survival through Activation of PI3K/Akt, ERK1/2 and Wnt/[beta]-Catenin Signaling Riccarda Granata, Eleonora Gargantini, Alessandra Baragli, Fabio Settanni, Marina Taliano, Ezio Ghigo University of Turin, Turin, Italy Obestatin is a recently discovered peptide encoded by the ghrelin gene. Obestatin was initially claimed to bind to the orphan receptor GPR39; however, this finding has been question and at present, obestatin receptor is still unknown. We have previously shown that obestatin exerts antiapoptotic effects in pancreatic [beta]-cells and human pancreatic islets, through activation of survival pathways and interaction with the glucagon-like peptide 1 receptor (GLP-1R). Besides peripheral actions, obestatin displays central effects, such as regulation of sleep and mnemonic functions. Hippocampal neurogenesis, which is essential for mnemonic functions and learning, consists of progenitor cell proliferation and differentiation. Interestingly, both ghrelin and the synthetic peptidyl growth hormone secretagogue hexarelin have been previously shown to stimulate proliferation of adult rat hippocampal progenitor cells (AHPs). Here, we investigated obestatin effects on proliferation and apoptosis of AHPs and the underlying signalling pathways. Cell survival was assessed by MTT assay, cell proliferation by 5-bromo-2-deoxyuridine (BrdU) incorporation, and apoptosis through caspase-3 activity and Bcl-2 expression. Obestatin increased cell proliferation and survival and reduced apoptosis of AHPs that were cultured in growth factor-deprived medium. These effects involved increased activity of phosphatidylinositol 3-kinase (PI3K)/Akt, extracellular signal-related kinase (ERK1/2), Wnt/[beta]-catenin and mammalian target of rapamycin (mTOR) signalling, as demonstrated by Western blot analysis and use of specific inhibitors. Furthermore, the specific antagonist of GLP-1R exendin-(9-39) abolished obestatin-induced proliferation and survival, suggesting GLP-1R-mediated signalling. These results indicate that obestatin promotes survival and proliferation and inhibits apoptosis of AHPs through activation of pathways which play a key role in neuroprotection. In addition, they suggest a possible role of obestatin in neuronal precursor cell protection and candidate this peptide as potential therapeutic molecule in conditions such as hippocampal damage.[br][br]Nothing to Disclose: RG, EG, AB, FS, MT, EG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 248 67 602 SAT-513 PO20-02 Saturday 541 2012


542 ENDO12L_SAT-514 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Knockout of Secretin in Purkinje Cells Changes Mouse Motor and Balance Behaviors Li Zhang, Billy KC Chow The University of Hong Kong, Hong Kong, Hong Kong Backgrounds: Secretin has been well established as a gut peptide hormone. Recently, the role of secretin in central nervous system has drawn lots of research interests. Previous studies demonstrated that secretin can modulate the inhibitory input to Purkinje cells in cerebellar cortex in rats(1). As Purkinje cells are responsible for controlling body motor and balance functions(2), in this study, we hypothesized that secretin in Purkinje cells plays a role in locomotor activities and balance abilities in mice.[br]Methodology: The Loxp-Secretin transgenic mice were mated with Pcp2-Cre transgenic line, in which the Cre-recombinase was expressed only in Purkinje cells. Offspring mice therefore had secretin expression removed in Purkinje cells but intact secretin levels in other brain sites as well as peripheral tissues, creating a Purkinje secretin knockout mouse model. Immunohistochemistry (IHC) staining was used initially to investigate and confirm secretin expression in conditional knockout mice. A full battery of behavior tests, focusing on the body motor function and balance abilities were employed, including open-field, rota-rod, wire-hang and vertical climbing tests. All tests were conducted on age-matched (month 2 to 9) littermates of conditional knockout and wildtype mice. Also knockout and wildtype mice were compared by the occurrence of a set of established neuronal reflex in the early neuronal developments.[br]Results: In Purkinje secretin knockout mice, secretin expression level was minimal in Purkinje cells but intact in other brain sites and peripheral organs as shown in the IHC staining. For the behavior tests, movement speed was intact in Purkinje secretin knockout mice compared to wildtype mice. The balance ability and fine motor control were however impaired in knockout mice, as shown in the rota-rod, wire-hang and vertical climbing test (p[lt]0.05). Such behavior and abnormalities occurred as early as in month 2 and persisted in all age groups checked (month 3 to 9). In addition, the knockout mice also showed late occurrence of neuronal reflex related to body motor functions and balance abilities (p[lt]0.05).[br]Conclusions: Secretin in Purkinje cells has effects in controlling mouse motor behaviors and body balance functions.[br][br]1. Yung WH, Leung PS, Ng SS, Zhang J, Chan SC, Chow BK. Secretin facilitates GABA transmission in the cerebellum. J Neurosci. 2001 Sep 15; 21(18):7063-8. 2. Sacchetti B, Scelfo B, Strata P.The cerebellum: synaptic changes and fear conditioning. Neuroscientist. 2005 Jun; 11(3): 217-27.[br][br]Nothing to Disclose: LZ, BKCC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 103 67 603 SAT-514 PO20-02 Saturday 542 2012


543 ENDO12L_SAT-515 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Accelerated Myelinogenesis in Galanin Transgenic Mice: Effect of Lipid Metabolism Lin Zhang, Shirley Hasdan, Jiming Kong, Maria Vrontakis University of Manitoba, Winnipeg, Canada Myelinogenesis is a scheduled process that depends on both the intrinsic properties of the cell and extracellular signals. Myelin development is mainly a postnatal event and lipids accumulate in the myelin during development. The relative molar proportions of these lipids (cholesterol, phospholipid, and galactolipids) change with age and are an index of myelin development. Lipids constitute an important part of myelin and significantly influence its integrity. Using our galanin transgenic mouse model, we have previously demonstrate that chronic increase of the neuropeptide galanin has pronounced neuroprotective effects with respect to demyelination and remyelination (1). There is also strong evidence that galanin is synthesized in myelin producing glial cells. In this transgenic mouse model we have also shown that the increase of circulating galanin levels induces marked alterations in lipid metabolism and increased serum levels of cholesterol in the absence of an orexigenic effect(2). In the present study we examined the effect of galanin on myelinogenesis in the CNS. Galanin transgenic mice at 10 and 20 postnatal days had accelerated myologenesis as it was demonstrated by the abundance of Basic Myelin Protein (MBP) and Uridine Diphosphate Glycosyltransferase 8 (UGT8), an enzyme catalyzing the transfer of galactose to ceramide, which is a key enzymatic step in the biosynthesis of galactocerebrosides, as well as high activated AMPK pathways, the signaling cascades involved in metabolic homeostasis and lipid metabolism of myelin formation, in their brains. These findings might indicate that high lipid metabolism in the myelinating environment of the galanin transgenic mice influence their myelinating process. Thus, besides a direct effect of increased galanin levels on preventing oligodendrocyte cell death, altered lipid metabolism due to increased galanin levels have an effect on myelinogenesis.[br][br](1)Zhang L et al.,PlosOne accepted. (2)Poritsanos N et al., Int J Obes 2009;33:1381-9.[br][br]Sources of Research Support: Manitoba Institute of Child Health, MS Society of Canada.[br][br]Nothing to Disclose: LZ, SH, JK, MV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 444 67 604 SAT-515 PO20-02 Saturday 543 2012


544 ENDO12L_SAT-516 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Characterization of Vasotocin Hormone and Two Putative Receptors in the Sea Lamprey ([italic]Petromyzon marinus[/italic]) Sally A Mayasich, Benjamin L Clarke University of Minnesota Medical School Duluth, Duluth, MN The sea lamprey (Petromyzon marinus) is an ancient jawless vertebrate that links jawed-vertebrates to non-vertebrates. This key evolutionary link is at the cusp of radical morphological establishment of a spine and jaw, and physiological changes expanding the immune and neuroendocrine systems. Vasopressin/vasotocin homologs and their cognate G-protein-coupled receptors have been characterized in bony fish through humans. However, little is known of lampreys, the most primitive vertebrate utilizing the vasotocin hormone. Basic Local Alignment Search Tool (BLAST) queries of the sea lamprey draft genome (The Genome Institute at Washington University School of Medicine in St. Louis, ftp://genome.wustl.edu/pub/organism/Other_Vertebrates//Petromyzon_marinus/assembly/Petromyzon_marinus-3.0/) and reverse transcription-polymerase chain reaction (RT-PCR) have resulted in amplification of the vasotocin hormone precursor on contig 19024.2 and two putative receptors possessing all seven transmembrane domains on contigs 2383.5 and 3198.5. The vasotocin hormone precursor translated region consists of 471 base pairs (bp). The 2383.5 receptor translated region consists of 1266 bp and contains one 4528-bp intron, and the 3198.5 receptor translated region consists of 1182 bp and contains a 2379-bp and a 5838-bp intron. Other partial receptor sequences that contained some transmembrane domains but not all seven were found in the lamprey genome database. Analysis of the sequence alignments and phylogenetic relationships among vertebrate species has revealed that the sea lamprey[apos]s two receptors are related based on sequence and structure to the V1b- and V1a-type vasopressin receptors as well as oxytocin-type receptors, but reside outside of these clades. No full receptor sequences were found that were homologous to the V2-type receptor which functions in anti-diuresis in higher vertebrates; however, tissue expression distribution of the 2383.5 and 3198.5 receptors is similar to that in higher vertebrates, indicating homology of physiological functions. In addition to canonical functions in the brain, gills and kidney, the expression of vasotocin receptor RNA in the pineal organ points to a role for this hormone in circadian rhythms and in the eye and olfactory sac in sensory processing; vasotocin hormone precursor RNA expression in white blood cells indicates a role in immune function.[br][br]Nothing to Disclose: SAM, BLC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2321 67 605 SAT-516 PO20-02 Saturday 544 2012


545 ENDO12L_SAT-517 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Secretogranin III along with Carboxypeptidase E Facilitates the Sorting of POMC into the Regulated Secretory Pathway of Endocrine Cells Niamh X Cawley, Trushar Rathod, Sigrid S Young, Y Peng Loh Program in Developmental Neuroscience, NICHD, National Institutes of Health, Bethesda, MD Carboxypeptidase E (CPE) and secretogranin III (SgIII) are proteins found in the granules of the regulated secretory pathway (RSP) of neuroendocrine cells. CPE has been shown to be an important molecule involved in sorting POMC and several other prohormones to the RSP, whereas SgIII has been shown to sort chromogranin A (CgA) to the RSP via interaction with cholesterol. In CPE mutant mice where CPE is degraded, regulated secretion of adrenocorticotropin (ACTH) from the anterior pituitary was proposed to be the result of increased levels of SgIII in these mice that facilitated sorting of POMC to the RSP in the absence of CPE. Indeed, in pull-down experiments in vitro, SgIII was able to interact with proopiomelanocortin (POMC), although not as efficiently as CPE, suggesting that SgIII may act as an alternate sorting receptor to that of CPE, to facilitate trafficking of POMC to the RSP similar to its sorting role of CgA. To address the role of SgIII sorting of POMC to the RSP in the mouse anterior pituitary cell line, AtT20, we reduced the expression of SgIII, by stable expression of shRNA directed against mouse SgIII. SgIII was reduced at both the mRNA and protein levels by [sim]80%. In non-radioactive steady state secretion assays, POMC was secreted constitutively at an elevated rate in the SgIII knock-down cells compared to control cells and while ACTH was secreted in a regulated manner, its levels were reduced. In pulse chase secretion experiments, newly synthesized POMC was also secreted constitutively at an elevated rate compared to control cells and ACTH was made and secreted in a regulated manner although the relative levels of the newly synthesized ACTH were reduced. Sucrose density gradient fractionation of SgIII knock-down cells, but not control cells, showed an accumulation of POMC and CgA in a less dense fraction of vesicles that co-fractionated with Golgi markers. Increased CgA and POMC staining in the Golgi by immunocytochemistry corroborated these observations. These results suggest that reduction of SgIII prevents the normal trafficking pattern of POMC and CgA within the cell resulting in their accumulation in a compartment that exists just prior to sorting into the RSP, presumably the trans Golgi Network (TGN), and subsequent increased constitutive secretion. In double knock-down cells of SgIII and CPE, constitutive POMC secretion was further elevated, demonstrating a combined role of CPE and SgIII in the trafficking POMC in these cells.[br][br]Sources of Research Support: The Eunice Kennedy Shriver National Institute of Child Health and Human Development.[br][br]Nothing to Disclose: NXC, TR, SSY, YPL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2368 67 606 SAT-517 PO20-02 Saturday 545 2012


546 ENDO12L_SAT-518 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Saturated Fatty Acids and High-Fat Diet Feeding Increase Type 2 Deiodinase Activity in the Mediobasal Hypothalamus Monika Toth, Gabor Wittmann, Peter Egri, Balazs Gereben, Csaba Fekete, Ronald M Lechan Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary; Tufts Medical Center, Boston, MA; Tufts University School of Medicine, Boston, MA Tanycytes are specialized cells of glial origin that line the floor and ventrolateral walls of the third ventricle in the mediobasal hypothalamus and extend long, cytoplasmic processes into adjacent feeding-related centers of the arcuate nucleus. Recent studies in our laboratories demonstrated that tanycytes are highly responsive to inflammatory mediators and markedly increase type 2 iodothyronine deiodinase (D2) activity, the major enzyme in the brain involved in conversion of T4 to its most biologically active end-product, T3, in response to bacterial endotoxin (LPS) through a NF-[kappa]B mediated mechanism (1-4). In addition, we demonstrated by RNA-Seq analysis that tanycytes express genes for Toll-like receptors (TLR) 2 and 4 and its adaptor, MyD88, in addition to numerous cytokines (5). Since saturated fatty acids are known to signal through TLRs and activate NF-[kappa]B, we hypothesized that saturated fatty acids might also induce D2 enzymatic activity in tanycytes. To test this hypothesis, 660 pmol palmitate was infused through a cannula permanently implanted into the lateral ventricle of mice. A significant increase in D2 enzymatic activity was observed 3h following the infusion compared to artificial CSF-infused controls (Control vs Palmitate (fmol/h/MBH): 1.49[plusmn]0.27 vs 2.42[plusmn]0.27, p[lt]0.05). In addition, mice placed on a high fat diet (HFD) (45% fat) compared to a 10% fat control diet significantly increased D2 enzymatic activity in tissue extracts from the most medial part of the mediobasal hypothalamus after 6 weeks of HFD feeding (Control vs HFD (fmol/h/MBH): 0.70[plusmn]0.05 vs 0.91[plusmn]0.08, P[lt]0.05) when body weight gain was significantly greater than controls, with a trend for increased D2 activity after 4 weeks of the HFD. To determine whether feeding-related neurons in the arcuate nucleus could be influenced by increased tanycyte T3 production, double-labeling histochemistry was performed. TR[alpha]1 was present in the nucleus of practically all arcuate nucleus neurons containing POMC-IR or NPY mRNA. We conclude that saturated fatty acids and HFD feeding significantly increase D2 in tanycytes, and that the resulting focal increase in mediobasal hypothalamic T3 could affect feeding-related neurons in the arcuate nucleus. We propose that the combination of local hypothalamic tissue hyperthyroidism induced by tanycyte D2 activation and the innate immune characteristics of these cells could profoundly impact appetite and energy homeostasis in HFD fed animals.[br][br](1) Fekete et al, Endocrinology 2004; 145:1649. (2) Fekete et al Endocrinology 2005; 146:1357. (3) Sanchez et al, Endocrinology 2008; 149:4329. (4) Sanchez et al Endocrinology 2010; 151:3827. (5) Iyer et al, Abs of 93rd Annual Meeting of The Endocrine Society, 2011.[br][br]Sources of Research Support: OTKA K81845, EU FW7 Health-F2-2010-259772, Lend[uuml]let Award of the Hungarian Academy of Sciences and NIH DK-37021.[br][br]Nothing to Disclose: MT, GW, PE, BG, CF, RML 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2107 67 607 SAT-518 PO20-02 Saturday 546 2012


547 ENDO12L_SAT-519 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Effect of Growth Hormone Replacement Therapy on Cognition after Traumatic Brain Injury: A Follow-Up Study Walter M High, Dennis J Zgaljardic, Jordan Harp, Charles R Gilkison, Randall J Urban, Brent E Masel University of Kentucky College of Medicine, Lexington, KY; The Transitional Learning Center, Galveston, TX; University of Kentucky College of Medicine, Lexington, KY; The University of Texas Medicial Branch, Galveston, TX; The University of Texas Medical Branch, Galveston, TX; University of Kentucky College of Medicine, Lexington, KY; University of Kentucky College of Medicine, Lexington, KY Background: Hypopituitarism and growth hormone deficiency (GHD) is common following moderate to severe traumatic brain injury (TBI). GHD from etiologies other than TBI has long been associated with lower performance on cognitive measures, especially those that assess information processing speed and memory. Replacement of growth hormone (GH) in persons who are GHD has generally resulted in improvement in cognitive performance. We have recently reported the results (High et al. 2010) of a small randomized, double-blind, placebo controlled study of 23 persons with moderate to severe TBI. In that study, following randomization, 12 participants were treated with active rhGH for one year while 11 participants injected themselves with placebo for one year. Neuropsychological testing was conducted at baseline, 6 months, and 12 months. At the end of one year, the group treated with rhGH showed improvements on measures that assess information processing speed, verbal memory, and finger tapping speed compared to the group receiving placebo. In the current study, we report findings from an additional year of active rhGH treatment in a subgroup of these patients with TBI.[br]Methods: A subgroup (n=8) of the original 12 participants receiving active rhGH was treated and followed for an additional year. For this follow-up study, neuropsychological performance at 24 months was compared to performance at 12 months.[br]Results: The group treated with active rhGH for 24 months showed no additional gains in cognitive performance compared to performance at the end of the first year on rhGH. On some measures, the gains moderated somewhat from year 1 to year 2, although noted declines in cognitive performance over this period of time were not statistically significant.[br]Conclusions: Larger trials replacing GH in persons who are GHD following TBI are needed. The results from preliminary studies are encouraging but larger long term studies are needed.[br][br]Sources of Research Support: This work was partially funded by the generous support of the Moody Endowment. This study was conducted on the Clinical Research Center (GCRC) at the University of Texas Medical Branch at Galveston funded by a grant #M01-RR00073 from the National Center for Research Resources, NIH, USPHS from the National Center for Research Resources.[br][br]Nothing to Disclose: WMH, DJZ, JH, CRG, RJU, BEM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2178 67 608 SAT-519 PO20-02 Saturday 547 2012


548 ENDO12L_SAT-520 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Mild and Moderate Traumatic Brain Injuries (mTBI) Disrupt Hypothalamic-Pituitary-Adrenal Axis Feedback and Impair Fear Extinction in Male Mice Sibyl N Swift, Stephen Shannon, David L Carbone, Jonathon Skibo, Robert J Handa, T John Wu Uniformed Services University of the Health Sciences, Bethesda, MD; University of Arizona College of Medicine, Phoenix, AZ Traumatic brain injuries (TBI) are known to cause disturbances in the HPA axis response to stress. However, little is known pertaining to the mechanism through which TBI disrupts HPA axis feedback. We used a novel brain injury model called the high intensity focused ultrasound (HIFU) to deliver a non-impact blast-induced TBI to explore this relationship. Adult male C57Bl/6 mice were subjected to mild (LOW) and moderate (MOD) HIFU exposure (equivalent to approximately 100 kPa and 200 kPa, respectively). Animals were tested 24h post-injury for: HPA axis (CORT) response to a 20 min immobilization stress, corticotrophin-releasing hormone (CRH) and glucocorticoid receptor (GR) mRNA, and fear extinction behavior. The MOD group exhibited elevated basal CORT (68.1+/-4.2 ng/ml vs. 47.5 +/-3.3 ng/ml, respectively), but a blunted response to restraint vs. sham controls (SHAM) (85.3 +/-4.7 ng/ml vs. 110.0+/-7.1 ng/ml, respectively). There were no differences in basal CORT for LOW vs. SHAM, but the CORT response to stress was attenuated (p[lt]0.05) for LOW vs. SHAM at the end of the stressor (99.4+/-5.0 ng/ml vs. 110.0+/-7.1 ng/ml, respectively) and 20 min after the end of the stress (75.8+/-6.6 vs. 98.1+/-4.8 ng/ml, respectively). CORT was not different (p[gt]0.05) at later time-points. CRH mRNA levels for the MOD group were lower in the paraventricular nucleus (PVN), but higher in the amygdala (AMG) compared to SHAM. GR mRNA was lower in the PVN and hippocampus, but greater in the AMG for MOD compared to SHAM. There were no detectable differences between LOW and SHAM for either CRH or GR mRNA levels. Fear conditioning revealed that SHAM and TBI (LOW and MOD) mice increase (p[lt]0.05) freezing during conditioning (65+/-11% vs. 73+/1-6%, respectively). During extinction, levels of freezing remained elevated in TBI mice (68+/-4%) compared to SHAM (38+/-9%) (p[lt]0.05). Our data suggest that exposure to TBI disrupts normal HPA axis by attenuating the feedback. These results suggest a dysregulation of HPA axis feedback following TBI which may lead to a loss of fear extinction.[br][br]Sources of Research Support: DoD DMRDP#D61_I_10_J6_125 and NIH NS039951.[br][br]Nothing to Disclose: SNS, SS, DLC, JS, RJH, TJW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1905 67 609 SAT-520 PO20-02 Saturday 548 2012


549 ENDO12L_SAT-521 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Mineralocorticoid Receptor Is a Neuroprotective Factor and an Epileptogenic Regulator: Involvement of HCN Channel Damien Le Menuet, Mathilde Munier, Giulia Campostrini, Say Viengchareun, Marc Lombes Faculty Medicine Paris SUd, Le Kremlin Bic[ecirc]tre, France The mineralocorticoid receptor (MR) exerts important physiological functions in various organs including the central nervous system (CNS). MR is highly expressed in the hippocampus where it controls memorization (long term potentiation), stress responses and is involved in epileptogenesis in association with the glucocorticoid receptor (GR) (1). Within the CNS, the Hyperpolarization Cyclic Nucleotide-gated (HCN) channels act as cationic channels presenting a spontaneous rhythmic activity. They are involved in long term potentiation, dendritic integration, and epilepsy when mutated, overlapping at least in part the spectrum of MR central actions. HCN channels or pacemaker channels are also pivotal for the control of heart beating rate. We previously demonstrated that embryonic stem (ES) cell derived-cardiomyocytes overexpressing the human MR (P1.hMR) exhibited an accelerated beating frequency. That was associated with an increased HCN1 channel expression, providing the first link between HCN channel and MR signaling (2). Besides, neuronal differentiation of P1.hMR and wild type (WT) ES cells leads to fully differentiated neurons that expressed several neuronal markers ([beta]-tubulin III, MAP2, synaptophysin) (3) as well as neurotransmitter receptors for NMDA, dopamine, GABA and serotonin along with the HCN channels. This strategy already allowed us to identify MR as a pro-survival molecule that confers relative resistance to oxidative stress-induced neuronal apoptosis (4). Neuronal MR overexpression is associated with an increased expression of HCN1, 2 and 4 channels compared to WT neurons. Using another neuronal cell type derived from mouse hippocampus generated by targeted oncogenesis (5), we demonstrate that BZ cells express HCN1, 2 and 4, neuronal markers (MAP2, synaptophysin, NMDA receptor) and a high concentration of MR and GR at both mRNA and protein level, thus constituting a useful cell[ndash]based system to further investigate neuronal mineralocorticoid signaling. The molecular mechanisms by which MR regulates HCN channel expression and their corticosteroid hormone regulation are currently under investigation. This work should provide new insights in the role of MR in epileptogenesis, memorization as well as neuron survival opening new avenues in neurodegenerative diseases, seizure-induced neuron injury and other CNS pathological states.[br][br]1- Kumar et al, Psychoneuroendocrinology 2007. 2- Le Menuet et al, Cardiovasc Res 2010. 3- Munier et al, Endocrinology 2010. 4- Munier et al, Endocrinology 2012. 5- Le Menuet et al, J Biol Chem 2000.[br][br]Sources of Research Support: Grants from Inserm, Univ Paris-Sud and ANR 2012.[br][br]Nothing to Disclose: DLM, MM, GC, SV, ML 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 462 67 610 SAT-521 PO20-02 Saturday 549 2012


550 ENDO12L_SAT-522 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Regulation of Gene Expression of CRH-Related Peptides and Their Receptor and Crosstalk with Inflammatory Cytokines in the Rat Brain Caused by Experimental Heart Strain Takanori Motoki, Miki Itohisa, Ei-Ichi Wake, Takashi Urashima, Ichiro Miyata, Michihiro Yoshimura, Katsuyoshi Tojo, Hiroyuki Ida The Jikei University School of Medicine, Minato-ku, Japan; The Jikei University School of Medicine, Minato-ku, Japan; The Jikei University School of Medicine, Minato-ku, Japan The roles of neuropeptides such as the CRH family on the condition of heart strain remain unknown. Furthermore, to our knowledge, there have been no studies reporting the crosstalk in heart failure between CRH-related peptides and inflammatory cytokines in the central nervous system. In our study, the regulation of gene expression in the brain and heart of CRH-related peptides and inflammatory cytokines was investigated using model rats with experimental left ventricular heart loading. First, we surgically prepared 5 model rats(Ao-banding group) by banding transverse aorta on mechanical ventilation, and 5 sham-operated rats as control group. Four weeks later, animals were sacrificed after assessment of their left ventricular endo-diastolic pressure(EDP) and endo-systolic pressure (ESP) by microcatheterization. Then, mRNA levels of urocortin(Ucn)-2, -3, CRF-2[alpha], Interleukin-6(IL-6) and Suppressor of cytokine signaling-3 (SOCS-3) in the brain and heart were studied by real-time RT-PCR. Additionally, serum concentrations of Ucn-2, -3 and IL-6 of both the Ao-banding and control rats were measured by EIA kits.[br]In the Ao-banding group, serum Ucn-2, Ucn-3 and IL-6 levels were significantly elevated compared with controls. Quantitative RT-PCR demonstrated that Ucn-2 and Ucn-3 mRNA ezpression of the Ao-banding group was upregulated in stiatum, hippocampus, thalamus, hypothalamus. On the other hand, IL-6 mRNA of the Ao-banding group was greatly upregulated in hippocampus, lateral septum, hypothalamus, amygdala, pituitary gland and heart like Ucn-2 and Ucn-3 mRNAs. Conversely, the downregulation of SOCS-3 mRNA expression was observed in the almost same tissues of Ao-banding group as IL-6 mRNA.[br]These results indicate that endogenous Ucn-2 and Ucn-3 through CRF-R2[alpha] are critical molecular mediators of the physiological response to heart failure in the central nervous system as well as in the heart. Taken together, our data suggest that left ventricular heart loading induces the increase of inflammatory cytokines such as IL-6, and furthermore affects the negative feedback system of cytokine signalings in the brain.[br][br]Nothing to Disclose: TM, MI, E-IW, TU, IM, MY, KT, HI 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 405 67 611 SAT-522 PO20-02 Saturday 550 2012


551 ENDO12L_SAT-523 POSTER SESSION: Hypothalamic Development [amp] General Neuroendocrinology (1:30 PM-3:30 PM) Aging and Social Isolation Increase Susceptibility to Anxiety in Mice Lacking 5[alpha]-Reductase Type 1 Emma M Di Rollo, Dawn EW Livingstone, Brian R Walker, Ruth Andrew Queen[apos]s Medical Research Institute, University of Edinburgh, Edinburgh, UK Glucocorticoid excess causes increased anxiety and accelerated cognitive decline. Tissue concentrations of glucocorticoids are influenced by local metabolism. In liver and brain, 5[alpha]-reductase 1 (5[alpha]R1) converts corticosterone to A-ring reduced metabolites, which have a different spectrum of activities. We tested 5[alpha]R1-/- mice (5[alpha]R1-KO) for heightened anxiety and impaired cognition.[br]Female wild-type (WT) and 5[alpha]R1-KO littermates were housed singly (S) or in groups (G). At 6 months (young) and 14-15 months (aged), we tested anxiety (elevated plus maze, EPM, and open field; expressed as % time in anxiogenic zones), learning and memory (Y-maze and Morris water-maze). In aged mice we obtained pituitary brains for qPCR. Data are mean[plusmn]SEM (KO-S vs KO-G vs WT-S vs WT-G), compared by two-way ANOVA,*p[lt]0.05,**p[lt]0.01 vs matched WT, n=17/group.[br]5[alpha]R1-KO, only when singly housed and aged, had higher plasma corticosterone levels than group-housed mice and increased anxiety (eg EPM:5.19[plusmn]2.32**vs11.30[plusmn]1.83vs10.73[plusmn]2.02vs10.83[plusmn]3.13%). 5[alpha]R1-KO exhibited subtle impairment of cognition, achieving the same performance as WT in the water maze, but with greater deterioration with age (p=0.05), independently of housing. Consistent with increased CNS glucocorticoid signalling, in 5[alpha]R1-KO mice hypothalamic glucocorticoid receptor (GR) mRNA was reduced (1.94[plusmn]0.16*vs1.38[plusmn]0.12#vs2.49[plusmn]0.29vs1.68[plusmn]0.12), and pituitary CRHR1 increased (1.17*[plusmn]0,12vs1.35*[plusmn]0.17vs0.91[plusmn]0.19vs0.97[plusmn]0.10), while mineralocorticoid receptor, CRH and pituitary GR were unchanged.[br]Disruption of 5[alpha]R1 and ageing induces anxiety and adversely affects cognition, effects which are exaggerated by environmental stress. Although systemic differences in metabolism or altered conversion of other steroids may play a role, these behavioral changes are consistent with glucocorticoid excess within the brain.[br][br]Nothing to Disclose: EMDR, DEWL, BRW, RA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1906 67 612 SAT-523 PO20-02 Saturday 551 2012


552 ENDO12L_SAT-524 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Suppression of StAR Gene Expression by SMILE in Leydig Cells Jin-Woo Yang, Seung-Yon Lee, Keesook Lee Chonnam National University, Gwangju, Republic of Korea SMILE ([underline]SM[/underline]all heterodimer partner [underline]I[/underline]nteracting [underline]LE[/underline]ucine Zipper Protein) has been initially identified as a protein, which interacts with orphan nuclear receptor SHP (Small Heterodimer Partner). It has been reported to directly interact with some nuclear receptors and regulate their transactivation in a cell-type specific manner. In this study, we investigated a role of SMILE in the expression of steroidogenesis-related genes in Leydig cells. SMILE protein is expressed in whole mouse testis and several mouse Leydig cell lines including mLTC-1. Adenovirus-mediated SMILE overexpression decreased the expression of StAR (Steroidogenic Acute Regulatory protein), one of steroidogenesis-related genes, in Leydig cells. In addition, promoter-reporter assays using StAR-luc revealed that SMILE suppressed the cAMP- and Nur77-mediated StAR promoter activity. Nur77 (NGFI-B[bold]) [/bold]is one of major transcription factors that are involved in the regulation of StAR gene expression and induced by cAMP in Leydig cells. Further reporter analysis using NBRE-luc showed that SMILE inhibited the transactivation of Nur77. Moreover, testosterone level of Ad-SMILE infected mLTC-1 cells were lower than Ad-GFP virus infected cells, indicating that SMILE inhibited testosterone synthesis in Leydig cells. Meanwhile, insulin resistance and metabolic syndrome have been suggested as risk factors for hypogonadism, and SMILE has been reported to be induced by insulin/AKT signaling in hepatocytes. In this study, we found that insulin also increased the SMILE protein level in Leydig cells. Taken together, these results suggest that insulin increases SMILE protein level and SMILE decreases the StAR gene expression by inhibiting Nur77 transcriptional activity, consequently decreasing the steroidogenesis in testicular Leydig cells.[br][br]Nothing to Disclose: J-WY, S-YL, KL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 302 68 613 SAT-524 PO25-01 Saturday 552 2012


553 ENDO12L_SAT-525 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Transforming Growth Factor-[beta]1 Represses Testicular Steroidogenesis Via Inhibition of Nur77 Transactivation Eunsook Park, Keesook Lee Chonnam National University, Gwangju, Republic of Korea Transforming growth factor-[beta]1 (TGF-[beta]1) has been reported to be an autocrine factor that inhibits LH mediated-steroidogenesis in testicular Leydig cells. However, the mechanism by which TGF-[beta]1 controls the steroidogenesis in Leydig cells is not well understood. In this study, we investigate the effect of TGF-[beta]1-activated Smad3 on the Nur77 transactivation in mouse Leydig cells. Nur77 is one of major transcription factors involved in steroidogenesis-related gene regulation in Leydig cells. In MA-10 cells, a mouse Leydig cell line, the overexpression of ALK5 TD, a constitutively active TGF-[beta] type I receptor, repressed the transactivation of Nur77 on the promoter of steroidogensis-related genes such as StAR, CYP17 and 3[beta]HSD. In addition, the repression of Nur77 transcactivation was increased by the co-overexpression of ALK5 TD and Smad3. Knockdown studies using siRNA revealed that Nur77 transactivation was repressed by endogenouse Smad3, but not by Smad2, and that Smad3-mediated repression of cAMP-induced CYP17 gene activation was through working on Nur77. Smad3-mediated repression of Nur77 transactivation occurred through the interference with the binding of Nur77 to DNA. Moreover, TGF-[beta]1 treatment inhibited cAMP-induced StAR, CYP17 and 3[beta]HSD gene expression in mature as well as immature primary Leydig cells. Taken together, these results suggest that TGF-[beta]1-activated Smad3 represses the expression of steroidogenic enzyme genes in mouse Leydig cells via the suppression of Nur77 transactivation. These findings may provide the molecular mechanism for the repression of testicular steroidogenesis by TGF-[beta]1.[br][br]Nothing to Disclose: EP, KL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 250 68 614 SAT-525 PO25-01 Saturday 553 2012


554 ENDO12L_SAT-526 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) ERR[gamma] Suppresses Expression of Steroidogenic Enzyme Genes Via Down-Regulation of Nur77 Jeong-Eun Seo, Eunsook Park, Keesook Lee Chonnam National University, Gwangju, Republic of Korea The orphan nuclear receptor Nur77 plays an important role in testicular steroidogenesis. Nur77, of which expression is induced by LH/cAMP signaling in testicular Leydig cells, activates the promoter of steroidogenic enzyme genes such as P450c17, StAR and 3[beta]HSD. In this study, we examined a role of estrogen-related receptor gamma (ERR[gamma]) in testicular steroidogenesis. ERR[gamma], an orphan nuclear receptor, has been known to be a constitutively active transcriptional activator. The overexpression of ERR[gamma], which is naturally expressed in mouse testis and Leydig cell lines, inhibited the expression of steroidogenic enzyme genes such as StAR and 3[beta]HSD. Promoter-reporter analyses showed that ERR[gamma] suppressed cAMP- or Nur77-induced activation of Nur77 target promoters including the natural promoter of steroidogenic enzyme genes. Further studies revealed that ERR[gamma] overexpression inhibited the cAMP-induced promoter activity of Nur77, decreasing Nur77 expression. Interestingly, ERR[gamma] co-expression also decreased the protein level of exogenously expressed Nur77, suggesting ERR[gamma] effect on Nur77 protein stability. Taken together, these results suggest that ERR[gamma] may suppress the Nur77-induced expression of steroidogenic enzyme genes primarily through the decrease of Nur77 protein level by gene regulation and protein destabilization in testicular Leydig cells.[br][br]Nothing to Disclose: J-ES, EP, KL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 301 68 615 SAT-526 PO25-01 Saturday 554 2012


555 ENDO12L_SAT-527 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) cAMP Signaling Regulates DGK[theta] Gene Expression in H295R Adrenocortical Cells Kai Cai, Marion Sewer University of California, San Diego, San Diego, CA In the human adrenal cortex, the biosynthesis of cortisol and adrenal adrogens is regulated by the peptide hormone adrenocorticotropin (ACTH). ACTH induces the intracellular production of cAMP and activation of the cAMP-dependent protein (PKA) kinase. DGK (diacylglycerol kinase) [theta] is an intracellular lipid kinase that phosphorylates diacylglycerol to form phosphatidic acid (PA). We have previously shown that PA is a ligand for the nuclear receptor steroidogenic factor 1 (SF1) and that cAMP signaling increases nuclear PA production. In this study, we sought to investigate the role of cAMP signaling in regulating on DGK [theta] gene expression. Real time RT-PCR analysis showed mRNA expression of DGK [theta] was increased in response to dibutyryl cAMP (dbcAMP). Western blot analysis revealed that dbcAMP increased DGK [theta] protein expression in the nucleus of H295R human adrenocortical cells. SF1 increased the transcriptional activity of a reporter plasmid containing 1.5kb of the DGK [theta] promoter fused to the luciferase gene. To identify the SF1 binding is required for DGK [theta] promoter activity, we mutated several putative SF-1 binding sites and found that multiple sites were required for SF1-dependent transcription. Consistent with this finding, chromatin immunoprecipitation (ChIP) assay demonstrated that dbcAMP signaling increased the recruitment of SF1 to the DGK[theta] promoter. Taken together, our findings suggest that SF1 activates DGK [theta] transcription in a cAMP-dependent manner in human adrenal cortex cells.[br][br]Nothing to Disclose: KC, MS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2276 68 616 SAT-527 PO25-01 Saturday 555 2012


556 ENDO12L_SAT-528 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) p21-Activated Kinase 6 (PAK6) Is a Novel Regulator of Steroidogenic Factor 1 (SF-1) and Steroidogenic Gene Transcription in H295R Human Adrenocortical Cells Natasha C Lucki, Donghui Li, Marion B Sewer University of California San Diego, La Jolla, CA p21-activated kinase (PAK) 6 is a serine/threonine kinase that belongs to group II of the PAK family. It contains a putative amino-terminal cdc42/Rac interactive binding motif and a carboxyl-terminal kinase domain. PAK6 overexpression has been implicated in prostate cancer cell growth, and targeted disruption of PAK6 revealed important roles for this kinase in locomotion and learning. PAK6 interacts with the androgen receptor and estrogen receptor alpha and interaction of the kinase with these steroid receptors inhibits the ability of the receptors to activate gene transcription. Unlike group I PAKs, which are regulated by an auto-inhibitory domain, the activity of PAK6 is regulated by phosphorylation. In particular, phosphorylation of PAK6 at serine 560 (Ser-560) activates the kinase and this phosphorylated form of PAK6 is highly expressed in multiple cancer cell lines. In the adrenal cortex, glucocorticoid production is primarily under the control of adrenocorticotropin (ACTH), which upon binding to its receptor activates PKA and leads to rapid cholesterol mobilization and the transcriptional activation of steroidogenic genes. Based on the fact that Ser-560 is a putative PKA site and the cAMP signaling pathway plays a key role in regulating steroidogenesis, we sought to define the role of PAK6 in regulating glucocorticoid biosynthesis in the human adrenal cortex. We show that PAK6 is rapidly activated in response to cAMP signaling in the nuclei of H295R cells. Interestingly, PAK6 interacts with steroidogenic factor 1 (SF-1), a nuclear receptor that regulates the transcription of most steroidogenic genes, and stimulates SF-1-dependent CYP17 reporter gene activity. Silencing PAK6 alters cAMP-dependent expression of multiple genes involved in steroid hormone biosynthesis and results in the accumulation of cholesterol in cells. Collectively, these studies identify PAK6 as a novel regulator of SF-1 function, steroidogenic gene expression, and glucocorticoid production.[br][br]Nothing to Disclose: NCL, DL, MBS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2279 68 617 SAT-528 PO25-01 Saturday 556 2012


557 ENDO12L_SAT-529 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) On the Role of HSD3B2 in Human Androgen Biosynthesis Sameer Udhane, Petra Kempna, Gaby Hofer, Primus E Mullis, Christa E Fluck University Children[apos]s Hospital Bern, Bern, Switzerland [bold]Background:[/bold][br]3beta-hydroxysteroid dehydrogenase (HSD3B) plays a crucial role in steroidogenesis. HSD3B2 is predominantly expressed in the adrenal cortex and the gonads while HSD3B1 is specifically found in the placenta and the peripheral tissues. HSD3B2 is differentially expressed during adrenal development and is overexpressed in PCOS ovaries. Human HSD3B2 mutations cause undervirilization in 46,XY and virilization in 46,XX newborns. However, little is known on HSD3B2 regulation. Previous studies identified some transcriptional regulators. Thus, this study aimed to further elucidate the HSD3B2 regulation.[br][bold]Methods/Results:[/bold][br]Studies were performed in cell models. In contrast to adrenal NCI-H295R cells, placental JEG3 and embryonic HEK293 cells do not express HSD3B2. To study transcriptional regulation of HSD3B2, cells were transfected with promoter luciferase reporters together with different transcription factors. We found that only transcription factors GATA4/6, SF1 and Nur77 together activated the HSD3B2 promoter in JEG3 and HEK293 cells. We also investigated whether HSD3B2 is regulated by epigenetic modification. Treatment with methyltransferase inhibitor 5-aza-[apos]2-deoxycytidine and histone deacetylase inhibitor trichostatin A were not stimulating HSD3B2 expression in JEG3 cell indicating that HSD3B2 is not regulated epigenetically. HSD3B2 activity was analyzed in adrenal NCI cells under different growth conditions and upon 8Br-cAMP stimulation. Serum starvation decreased HSD3B2 promoter activity and protein expression. Overall androgen (DHEA) production in NCI cells was increased by starvation. 8Br-cAMP stimulation increased HSD3B2 promoter activity through the Nur77 but not the GATA promoter cis-element. Interestingly, 8Br-cAMP stimulation diminished HSD3B2 activity within 24 hours indicating that HSD3B2 is not only regulated at the transcriptional level.[br][bold]Conclusion:[/bold][br]HSD3B2 is not regulated by epigenetic modification. However, HSD3B2 is strongly regulated at the transcriptional level by GATA4/6, SF1 and Nur77. Serum withdrawal shifts steroidogenesis of NCI cells towards androgen production by repressing HSD3B2 expression. Cyclic AMP stimulation enhances HSD3B2 promoter activity but decreases HSD3B2 activity within 24 hours. Interestingly, a similar cascade of events is described for the adrenal cortex during adrenarche when HSD3B2 expression drops in the zona reticularis and production of DHEA rises at around age 6-8 years.[br][br]Nothing to Disclose: SU, PK, GH, PEM, CEF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 500 68 618 SAT-529 PO25-01 Saturday 557 2012


558 ENDO12L_SAT-530 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Partial CYP11A1 Deficiency Due to R90K Mutation Manifesting as Adrenal Insufficiency Only without Affecting Pubertal Development in a 46,XX Subject Sameer Udhane, Nuria Camats, Bernhard Dick, Amit V Pandey, Monica Fernandez-Cancio, Brigitte Frey, Laura Audi, Primus E Mullis, Christa E Fluck University Children[apos]s Hospital Bern, Bern, Switzerland; University of Bern, Bern, Switzerland; Hospital Universitari Vall d[apos]Hebr[oacute]n, Barcelona, Spain [bold]Introduction:[/bold][br]CYP11A1 is the first and rate-limiting enzyme for all steroid hormone biosynthesis from cholesterol. So far, eleven CYP11A1 mutations have been described in two 46,XX and nine 46,XY subjects. All subjects presented with symptoms of adrenal insufficiency (AI) between birth and 9 years of age while seven 46,XY patients also manifested with moderate to severe undervirilization. Partial enzyme activity is thought to explain later onset of AI and normal sexual development. However, the impact of partial CYP11A1 deficiency on pubertal development remains unknown.[br][bold]Methods/Results:[/bold][br]We describe the impact of a novel CYP11A1 mutation in a 46,XX patient from birth to adulthood. The patient was born at term after a normal pregnancy but manifested at the age of 10 months with an adrenal crisis during gastroenteritis. She was treated with mineralocorticoids and glucocorticoids and developed normally except for overweight problems. At the age of 12 years she started puberty. Menarche was at 14 years with regular menstrual cycles to the actual age of 18.5 years. ACTH stimulation test at age 15 years (72 hours after last fludrocortisone and hydrocortisone dose) revealed grossly elevated plasma ACTH and renin at baseline and unmeasurable baseline adrenal steroids without reactivity to stimulation. At age 18 years, plasma LH, FSH, E2, AMH and inhibin B were all normal, but DHEA/-S were not measurable and androstenedione was low. Ultrasound revealed no adrenal glands, an adult shaped uterus and ovaries with a single cyst on the left. Genetic testing was performed for congenital primary adrenal insufficiency and a novel homozygote R90K CYP11A1 mutation was found. Parents were heterozygote carriers. The mutation was tested for its activity in a cell model using 22R-OH-cholesterol as substrate for the production of pregnenolone. R90K had partial activity. Structure-function studies were performed.[br][bold]Conclusions:[/bold][br]Overall, partially active CYP11A1 mutations seem to affect predominantly adrenal steroid production while gonadal steroidogenesis may remain normal, allowing for a normal pubertal development. Whether normal gonadal function sustains for normal fertility remains to be seen. Thus, partially active CYP11A1 mutations seem to have a similar clinical spectrum as partially active StAR mutations which may allow for normal pubertal development in both sexes but may have compromised fertility with age. Protective measures for the gonad may be discussed.[br][br]Nothing to Disclose: SU, NC, BD, AVP, MF-C, BF, LA, PEM, CEF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1979 68 619 SAT-530 PO25-01 Saturday 558 2012


559 ENDO12L_SAT-531 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Broad Phenotypic Spectrum of 17[alpha]-Hydroxylase Deficiency: Functional Characterization of 4 Novel Mutations in the [italic]CYP17A1[/italic] Gene Jan Idkowiak, Silvia Parajes, Savitha Shenoy, Vivek Dhir, Chankramath S Arun, Ewa M Malunowicz, Norman Taylor, Cedric H Shackleton, Guy T[apos]Sjoen, Tim Cheetham, Wiebke Arlt, Nils Krone University of Birmingham, Birmingham, UK; Leicester Royal Infirmary, Leicester, UK; Royal Victoria Infirmary, Newcastle-upon-Tyne, UK; The Children[apos]s Memorial Health Institute, Warsaw, Poland; King[apos]s College Hospital, London, UK; University of Gent, Gent, Belgium The steroid 17[alpha]-hydroxylase enzyme CYP17A1 exerts two distinct activities that catalyze conversion reactions at key branch points in steroidogenesis. CYP17A1 17[alpha]-hydroxylase activity is the key step in cortisol synthesis whereas CYP17A1 17,20 lyase activity generates sex steroid precursors. Inactivating [italic]CYP17A1[/italic] mutations result in CYP17A1 deficiency (17OHD), a rare form of congenital adrenal hyperplasia that classically presents with combined glucocorticoid and sex steroid deficiency and hypokalaemic hypertension.[br]We have investigated four patients with 17OHD harbouring four novel mutations, which we analyzed [italic]in vitro[/italic] and [italic]in silico[/italic]. Case 1 (46,XY; homozygous p.F53/54del) presented at 12 years with glandular hypospadias, gynaecomastia and cryptorchidism. Endocrine assessment revaled hypergonadotropic hypogonadism, low cortisol at baseline and after cosyntropin stimulation, but notably normal blood pressure. Case 2 (46,XX; homozygous p.Y60IfsK88X) presented with severe adrenal insufficiency three weeks after birth. Case 3 (46,XX; p.G111V/p.P409L) presented at the age of 15 years with lack of pubertal development and a history of hypokalaemic hypertension since the age of 2 years. Case 4 (46,XY; p.R347H/p.A398E) was born with ambiguous genitalia but had normal blood pressure and no evidence of glucocorticoid deficiency at the age of 24 years suggesting isolated 17,20 lyase deficiency. Urinary steroid profiling with gas chromatography/mass spectrometry established the biochemical diagnosis of 17OHD in all cases. Functional [italic]in vitro[/italic] analysis of CYP17A1 activities in transiently transfected Cos1 cells confirmed p.Y60IfsK88X and p.P409L to abolish CYP17A1 function; p.A398E, p.F53/54del and p.R347H resulted in mild to moderate impairment of enzyme activity. Results of [italic]in silico[/italic] analysis of the identified mutations were consistent with the [italic]in vitro[/italic] findings.[br]In summary, we have identified four novel [italic]CYP17A1[/italic] mutations and our functional studies confirmed the pathogenicity of these mutations. The clinical presentations ranged from neonatal presentation with severe adrenal insufficiency to delayed pubertal development and normal adrenal function, illustrating the broad phenotypic spectrum in 17OHD.[br][br]Nothing to Disclose: JI, SP, SS, VD, CSA, EMM, NT, CHS, GT, TC, WA, NK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1600 68 620 SAT-531 PO25-01 Saturday 559 2012


560 ENDO12L_SAT-532 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Unrecognized Activities and Mechanistic Insight for the Human Steroid Hydroxylases CYP17A1 (P450c17) and CYP21A2 (P450c21) Derived from Metabolism of Deuterium-Labeled Steroid Substrates Francis K Yoshimoto, Yishan Zhou, David K Stidd, Hwei-Ming Peng, Jennifer A Yoshimoto, Kamalesh K Sharma, Richard J Auchus University of Texas Southwestern Medical Center, Dallas, TX; University of Michigan, Ann Arbor, MI Cortisol biosynthesis requires the activities of steroid hydroxylases CYP17A1 (P450c17, 17-hydroxylase/17,20-lyase) and CYP21A2 (P450c21, 21-hydroxylase). These microsomal P450 enzymes catalyze hydroxylation of progesterone and related steroids at sites within a 1[Aring] radius. We probed their mechanisms of hydrogen atom abstraction and hydroxylation at alternate sites with selectively deuterium-labeled substrates.[br]We synthesized 17-d[sub]1[/sub]-pregnenolone, 17-d[sub]1[/sub]-progesterone, 16[alpha]-d[sub]1[/sub]-progesterone, 21,21,21-d[sub]3[/sub]-progesterone, and 21,21,21-d[sub]3[/sub]-17-hydroxyprogesterone. We incubated these steroids with yeast microsomes containing human P450-oxidoreductase and wild-type human CYP17A1 or mutation A105L[mdash]with low progesterone 16[alpha]-hydroxylase activity[mdash]and wild-type human CYP21A2 or mutation V359A, hydroxylates progesterone at the 21- and 16[alpha]-positions in a 3:2 ratio. We quantified the products and calculated intramolecular kinetic isotope effects (KIEs). We also performed competition studies with [[sup]3[/sup]H]-labeled steroids to determine competitive intermolecular KIEs.[br]We found low intramolecular KIEs for CYP17A1 and mutation A105L at H-17 for all 17-hydroxylations (1.3-1.6) and larger KIEs at H-16[alpha] (2.5-4.8). Mutation A105L 21-hydroxylates progesterone (5% of products), and wild-type CYP17A1 also catalyzes 2% 21-hydroxylation, which is increased to 5% upon incubation with 16[alpha]-d[sub]1[/sub]-progesterone. CYP17A1 also 16[alpha]-hydroxylates 17-d[sub]1[/sub]-pregnenolone ([ldquo]metabolic switching,[rdquo] 5%) but not pregnenolone. Incubations with CYP21A2 mutation V359A and 16[alpha]-d[sub]1[/sub]- or 21,21,21-d[sub]3[/sub]-progesterone yielded intramolecular KIEs of 3.0-3.3. Wild-type CYP21A2 also forms a trace of 16[alpha]-hydroxyprogesterone, which increased to 4% with 21,21,21-d[sub]3[/sub]-progesterone substrate. Incubations with wild-type CYP21A2 or mutation V359A and 21,21,21-d[sub]3[/sub]-17-hydroxyprogesterone also demonstrated metabolic switching to partially-characterized products. Intermolecular KIEs paralleled the intramolecular KIE values.[br]We conclude that human CYP17A1 has progesterone 21-hydroxylase activity and that human CYP21A2 has progesterone 16[alpha]-hydroxylase activity, both of which are enhanced and revealed with deuterated substrates. The transition state for C-H bond cleavage in the 17-hydroxylase reaction is either significantly non-linear and/or asymmetric, and C-H bond breakage is partially rate-limiting for all reactions. These findings might be exploited to engineer additional activities to these P450s to treat deficiencies of another enzyme.[br][br]Sources of Research Support: R01-GM08659602 from NIGMS.[br][br]Nothing to Disclose: FKY, YZ, DKS, H-MP, JAY, KKS, RJA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2343 68 621 SAT-532 PO25-01 Saturday 560 2012


561 ENDO12L_SAT-533 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Potent Inhibitor of 17[beta]-Hydroxysteroid Dehydrogenase Type 1 Causes Changes in Metabolite Profile in Breast Cancer Cell Lines as Revealed by Targeted Metabolomics Pauline Banachowicz, Anna Halama, Emanuel Bey, Cornelia Prehn, Gabriele Moeller, Rolf W Hartmann, Jerzy Adamski Helmholtz Zentrum Muenchen, Neuherberg, Germany; Saarland University, Saarbruecken, Germany Breast cancer is the second most common cause of death among women in western countries. Overexpression of 17beta-hydroxysteroid dehydrogenase type 1 (17beta-HSD1), an enzyme involved in estradiol biosynthesis, is associated with bad prognosis in cancer disease. Inhibition of 17beta-HSD1 is a promising therapeutic approach and many specific modulators of this enzyme have been developed so far. Recently, a potent fluorine non-steroidal inhibitor of 17beta-HSD1 was described (1). We applied this compound, and the less potent carbenexolone, to two different cancer cell lines (T47D and MCF-7) expressing 17beta-HSD1 and evaluated the impact on the target enzyme as well as off-target processes. Cell proliferation, viability analyses and activity assays were performed after 6 h, 24 h and 48 h of incubation with inhibitors in steroid-free media. We further performed metabolomics analyses of treated cells and conditioned media to evaluate non-target effects of the inhibitors. For characterization of changes in the metabolite profile were measured a metabolite panel consisting of amino acids, acylcarnitines, phospholipids, hexoses and sphingomyelins by high throughput LC-MS/MS targeted metabolomics. Significant changes in metabolite profile of challenged cells were found.[br][br](1) Bey E. et. al, J. Med. Chem. 2009, 52, 6724[ndash]6743.[br][br]Sources of Research Support: Deutsche Forschungsgemeinschaft (DFG) grants to RWH (HA 1315/12-1) and JA (AD 127/10-1).[br][br]Nothing to Disclose: PB, AH, EB, CP, GM, RWH, JA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 55 68 622 SAT-533 PO25-01 Saturday 561 2012


562 ENDO12L_SAT-534 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Characterization of Type 15 17[beta]-Hydroxysteroid Dehydrogenase Melanie Samson, Fernand Labrie, Van Luu-The Laval University, Quebec, Canada; Laval University, Quebec, Canada; Centre de Recherche du CHUQ, Quebec, Canada Seventeen hydroxysteroid dehydrogenases (17[beta]-HSDs) represent a family of enzymes issued from convergent evolution of activity from unrelated ancestor genes. Accordingly, they share very low amino acid sequence identity (approximatedly 20-25%), except those belong to the same gene family. For example, types 3 and 12 17[beta]-HSD have common ancestor gene with long chain fatty acid elongation, type 5 17[beta]-HSD with aldo-keto reductases and types 6, 9 and 15 17[beta]-HSD have common ancestor gene with retinol dehydrogenases. Thus, 17[beta]-HSDs generally possess multiple activities, able to metabolize substrates of ancestor activities as well as those produced more recently. Their physiological role will depend on the local expression and availibility of precursors and receptors. A decade ago[bold][italic], [/italic][/bold]Lin et al. (1) report the sequence and expression of an enzyme named prostate specific dehydrogenase reductase 1 (PSDR1) which is highly stimulated by androgen in LNCaP. Subsequently, Kedishvili et al. (2) overproduced this enzyme in insect SF9 cells using baculovirus expression system and found that the microsomal preparation of infected cells does not metabolize steroids. However, using cultured intact HEK-293 cells stably expressing the enzyme, we have found that PSDR1 possesses 17[beta]-HSD activity catalyzing the transformation of androsterone (ADT) to 5[alpha]-androstane-3[alpha],17[beta]-diol and more importantly 5[alpha]-androstanedione into dihydrotestosterone (DHT). At a much lesser extent PSDR1 is also able to catalyze the transformation of 4-androstenedione into testosterone. Due to its 17[beta]-HSD activity, we, chronologically, named this enzyme type 15 17[beta]-HSD. The discrepancy between our results and those of Kedishvili et al. is unclear. It could be due to differences in post-modification system in insect and mammalian cells, or a difference in the enzymatic assays used, namely microsomes versus cultured cells. [italic]In situ [/italic]hybridization of normal prostate showed PSDR1 expression in both basal and luminal epithelial cells but not in fibromuscular stromal cells, endothelial cells, or infiltrating lymphocytes. By contrast, Psdr1 expression is very low in the mouse prostate (3) (approximately 50-fold lower than in human), thus suggesting very different functions of the enzyme in the human and mouse prostate.[br][br](1)Lin B et al.,Cancer Res 2001;61:1611. (2)Kedishvili et al., J Biol Chem 2002; 277:28909. (3)Moore et al., Gene 2002; 293:149.[br][br]Sources of Research Support: Endorecherche Inc.[br][br]Nothing to Disclose: MS, FL, VL-T 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 763 68 623 SAT-534 PO25-01 Saturday 562 2012


563 ENDO12L_SAT-535 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Analysis of Previous Data from 17[beta]-Hydroxysteroid Dehydrogenase Deficient Patients Suggests That Testosterone Is Not a Major Substrate for 5[alpha]-Reductases and Aromatase Van Luu-The Laval University, Quebec, Canada; Laval University, Quebec, Canada; Centre Hospitalier Universitaire de Quebec (CHUQ), Quebec, Canada The cloning and characterization of multiple androgen and estrogen specific 17[beta]-hydroxysteroid dehydrogenases (17[beta]-HSDs), together with higher affinity of 4-androstenedione (4-dione) for 5[alpha]-reductases and aromatase than testosterone (T), strongly suggest that the steps catalyzed by 5[alpha]-reductase and aromatase in steroidogenic pathways precede the step catalyze by 17[beta]-HSDs (4-dione ---5[alpha]-reductase---[gt] 5[alpha]-androstanedione (5[alpha]-dione) ----17[beta]-HSD---[gt] dihydrotestosterone (DHT); and 4-dione ---- aromatase --[gt] estrone (E1) --- 17[beta]-HSD ---[gt] estradiol (E2)). Such pathways are in contradiction with pathways generally described in the literature, in which circulating T, produced by gonads, is the substrate of aromatase and 5[alpha]-reductases. According to this pathway, the step catalyzed by 17[beta]-HSD precedes the steps catalyzed by aromatase and 5[alpha]-reductases, therefore only one or two 17[beta]-HSDs are necessary instead of 15 characterized. Using labeled-DHEA and inhibitors of 5[alpha]-reductase and aromatase, respectively, in cultured cell lines, we have shown recently that the steps catalyzed by 5[alpha]-reductases and aromatase precede the step catalyzed by 17[beta]-HSDs (1,2). To determine whether such pathways could be effective physiologically, we analyze previous data from 17[beta]-HSD deficient patients published by Saez et al.(3) and Rosler et al. (4). Saez et al. (1970) showed that although the level of T is decreased in these patients, their levels of 5[alpha]-reduced keto-steroids are high. In addition, Rosler et al. (1992) show that the level of DHT is higher in pseudohermaphroditism patients than in unaffected individuals. These data strongly suggested that 5[alpha]-reduced steroids are not derived from T. On the other hand, it has been reported that gynecomastia is generally found in patient having a deficient in 17[beta]-HSD (3). This is most probably due to an excess of E2 produced from the pathway that does not require T as an intermediate.We have quantified expression levels of several 17[beta]-HSDs and 5[alpha]-reductases in different tissues. Our data show low levels of 5[alpha]-reductase but high levels of type 3 and 5 17[beta]-HSDs in the testis and muscle, respectively, suggesting that T is the active androgen in these tissues. On the other hand, high levels of 5[alpha]-reductases have been found in the prostate, skin and brain suggesting that DHT, most probably produced from 4-dione by the pathway that does not require T as an intermediate, is the active androgen in these tissues.[br][br](1) Samson et al. J Invest Dermatol 2010;130:602. (2) Samson et al. J Steroid Biochem Mol Biol 2009;116:154. (3) Saez et al. J Clin Endocrinol Metab 1971;32:604. (4) Rosler et al. J Clin Endocrinol Metab 1992;75:773.[br][br]Sources of Research Support: Canadian Institute of Health Research to VLT.[br][br]Nothing to Disclose: VL-T 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1238 68 624 SAT-535 PO25-01 Saturday 563 2012


564 ENDO12L_SAT-536 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Crystal Structures of Human 17[beta]-Hydroxysteroid Dehydrogenase Type 5 (AKR1C3) in Complex with [italic]N[/italic]-Phenylanthranilic Acid and Indomethacin-Based Selective Inhibitors Mo Chen, Adegoke O Adeniji, Barry M Twenter, Andy J Liedtke, Jeffrey D Winkler, Lawrence J Marnett, David W Christianson, Trevor M Penning Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA; Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA; University of Pennsylvania, Philadelphia, PA; Vanderbilt University, Nashville, TN [italic]AKR1C3[/italic] is among the most up-regulated genes in castrate resistant prostate cancer (CRPC). The enzyme functions downstream in the androgen biosynthetic pathway by converting [Delta][sup]4[/sup]-androstene-3,17-dione to testosterone and 5[alpha]-androstane-3,17-dione to 5[alpha]-dihydrotestosterone. Inhibition of AKR1C3 may selectively prevent androgen biosynthesis in CRPC without perturbing adrenal steroidogenesis, a side effect of abiraterone. An ideal AKR1C3 inhibitor should not inhibit other related AKR1C isoforms involved in androgen metabolism. We have developed two series of analogs based on nonsteroidal anti-inflammatory drugs [italic]N[/italic]-phenylanthranilic acid and indomethacin as AKR1C3 inhibitors. Lead compounds in the two series that show nanomolar potency and over 100-fold selectivity for AKR1C3 have been submitted for structural studies. We have previously reported the crystal structure of the AKR1C3[bull]NADP[sup]+[/sup][bull]3-(4-(trifluoromethyl)phenylamino)benzoic acid complex ([bold]1[/bold]) (PDB# 4DBU), in which the inhibitor exhibits a similar binding pose to that of flufenamic acid (PDB# 1S2C) except its trifluoromethyl-phenyl ring penetrates deeper into the binding subpocket, SP1. Here we present the crystal structures of AKR1C3[bull]NADP[sup]+[/sup] in complex with two novel inhibitors, 3-(4[apos]-(nitronaphthalen-1-amino))benzoic acid ([bold]2[/bold], PDB# 4DBS) and 2[apos]-desmethyl-indomethacin ([bold]3[/bold], PDB# 4DBW) determined at 1.80 and 1.85 [Aring] resolution, respectively. Both inhibitors are anchored to the oxyanion site via their carboxylate groups. The naphthyl ring of [bold]2[/bold] and the [italic]p[/italic]-chlorobenzoyl ring of [bold]3[/bold] both extend into SP1 as observed with the trifluoromethyl-phenyl ring of [bold]1[/bold], however, both [bold]2[/bold] and [bold]3[/bold] penetrate this pocket more deeply. This penetration cannot be accommodated by the other AKR1C isoforms that have smaller subpockets. The common binding conformation shared by all three inhibitors confirmed that SP1 confers inhibitory selectivity for AKR1C3 and should be exploited for structural based rational drug design. [italic]In vitro[/italic] cell-based assays of compound [bold]2[/bold] indicate that in addition to acting as an AKR1C3 inhibitor it also functions as an androgen receptor antagonist, suggesting that the compound can be optimized as a bifunctional agent. The 2[apos]-desmethyl indomethacin complex structure shows that this analog binds differently to indomethacin and that this new binding pose can be used to improve analog synthesis. The biological properties of a series of indomethacin analogs are presented in a separate abstract submitted by Adeniji, AO, et al.[br][br]Sources of Research Support: The agents disclosed in this abstract are protected by US Provisional Patent Applications No. 61/465,091 and No. 61/548,004. Supported by NIH Grants R01-DK40715, R01-CA-90744, Prostate Foundation Challenge Grant P30-ES013508 and UL1RR024134 awarded to TMP; GM-056838 awarded to DWC; F32DK089827 awarded to M.C; and R01-CA889450 to LJM].[br][br]Nothing to Disclose: MC, AOA, BMT, AJL, JDW, LJM, DWC, TMP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 715 68 625 SAT-536 PO25-01 Saturday 564 2012


565 ENDO12L_SAT-537 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Development of Potent and Selective Indomethacin Analogs for the Inhibition of AKR1C3 in Castrate-Resistant Prostate Cancer Adegoke O Adeniji, Andy Liedtke, Michael C Byrns, Yi Jin, Larry J Marnett, Trevor M Penning Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA; Vanderbilt University School of Medicine, Nashville, TN Castrate resistant prostate cancer (CRPC), a fatal metastatic form of prostate cancer often develops in prostate cancer patients that were initially responsive to androgen deprivation therapy. CRPC is characterized by a reactivation of androgen receptor (AR) signaling often driven by elevated expression of enzymes involved in local androgen biosynthesis and consequently increased intratumoral androgen production, despite the existence of castrate levels of circulating androgens. [italic]AKR1C3 [/italic]also known as (Type 5 17[beta]-hydroxysteroid dehydrogenase) is among the most upregulated genes in CRPC and converts androgen precursors 4-androstene-3,17-dione and 5[alpha]-androstandione to the potent androgens, testosterone and 5a-dihydrotestosterone, respectively. The involvement of AKR1C3 in the pre-receptor regulation of AR action and its intratumoral localization makes it an important target in treatment of CRPC. Inhibitors of AKR1C3 should not inhibit the related isoforms, AKR1C1 and AKR1C2 since the latter enzymes are involved in the inactivation of 5[alpha]-dihydrotestosterone within the prostate.[br]Indomethacin used clinically to inhibit cyclooxygenase enzymes inhibits AKR1C3 potently and displays high selectivity over AKR1C1 and AKR1C2. We report the discovery of three classes of Indomethacin-based nanomolar inhibitors of AKR1C3 with over 100 fold selectivity over AKR1C1 and AKR1C2. The lead compounds are also devoid of inhibitory activity on cyclooxygenase enzymes and display robust inhibition of testosterone formation in a LNCaP-AKR1C3 prostate cancer cell line. Indomethacin has been shown to block PSA, ERG expression, and cell proliferation in a VCaP xenograft model of CRPC in an AKR1C3 dependent manner providing [italic]in vivo [/italic]efficacy data (Cai at al., Cancer Res. 71: 6503, 2011). An X-ray crystal structure of a lead compound (2[apos]-des-methyl-indomethacin) bound to the AKR1C3.NADP+ complex is presented as a separate abstract ([bold][italic]Chen et. al, Crystal structures of human 17[beta]-hydroxysteroid dehydrogenase type 5 (AKR1C3) in complex with N-phenylanthranilic acid and indomethacin based selective inhibitors.[/italic][/bold] These inhibitors are predicted to have comparable pharmacokinetic profiles to Indomethacin and have the potential to be therapeutic agents for the treatment of CRPC.[br][br](Cai at al., Cancer Res. 71: 6503, 2011).[br][br]Sources of Research Support: R01-CA97044, P30-ES013508, and a Prostate Cancer Foundation Challenge Award to TMP and by R01-CA889450 to LJM.[br][br]Nothing to Disclose: AOA, AL, MCB, YJ, LJM, TMP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 873 68 626 SAT-537 PO25-01 Saturday 565 2012


566 ENDO12L_SAT-538 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Biologically Active Form of 3-beta Hydroxysteroid Dehydrogenase (3[beta]HSD2) Is a Molten Globule Manoj Prasad, James L Thomas, Randy M Whittal, Himangshu S Bose Mercer University School of Medicine, Savannah, GA; Mercer University School of Medicine, Macon, GA; University of Alberta, Edmonton, Canada; Mercer University School of Medicine, Savannah, GA Inner mitochondrial membrane protein 3-beta hydroxysteroid dehydrogenase2 (3[beta]HSD2) synthesizes progesterone and androstenedione through its dehydrogenase and isomerase activities. This bifunctionality requires 3[beta]HSD2 to undergo a conformational change but its mechanism of action is unknown. Given its proximity to the mitochondrial proton pump, we hypothesized that pH influences 3[beta]HSD2 conformational changes associated with activity. Circular dichroism (CD) showed that between pH 7.4 and 4.5, 3[beta]HSD2 retained its primarily [alpha]-helical character with a decrease in [alpha]-helical content at lower pHs, but [beta]-sheet content remained unchanged. Titrating the pH back to 7.4 restored the original conformation. Pregnenolone to progesterone conversion indicated maxmum 3[beta]HSD2 activity at pH 4.5 than at pH 3.5 and 7.4. Increasing 3[beta]HSD2 concentration from 1[mu]g to 40 [mu]g resulted in a 7-fold increase in progesterone at pH 4.5, but no change at the physiological pH. Incubation with chaotropic Guanidinum Hydrochloride (GnHCl) resulted a three-step unfolding of 3[beta]HSD2 from pH 7.4 to pH 4.5, possibly due to the native state unfolding to the intermediate ion core state. With further decreases in pH, increasing concentrations of GnHCl led to rapid two-step unfolding representing complete loss of structure. Between pH 4 and 5, the two intermediate states was stable. Stopped-flow kinetics showed slower unfolding at around pH 4, where the protein is in a pseudostable state. Based on our data, we conclude that at pH 4 to 5, 3[beta]HSD2 takes on a molten globule conformation that promotes the dual functionality of the 3[beta]HSD2.[br][br]Sources of Research Support: National Institutes of Health HD057876 to HSB.[br][br]Nothing to Disclose: MP, JLT, RMW, HSB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 257 68 627 SAT-538 PO25-01 Saturday 566 2012


567 ENDO12L_SAT-539 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Structural and Functional Implications of the Aromatase Oligomer Jessica Lo, Giovanna Di Nardo, Jennifer Griswold, Wenhua Jiang, Gilardi Gianfrano, Debashis Ghosh State University of New York (SUNY) Upstate Medical University, Syracuse, NY; University of Torino, Torino, Italy; Hauptman- Woodward Medical Research Institute, Buffalo, NY Aromatase, a member of the cytochrome P450 superfamily, is a steroidogenic enzyme responsible for the conversion of androgens to estrogens. Estrogen overproduction in the peripheral tissue is responsible for 75% of breast cancer in postmenopausal women. As a result, aromatase inhibitors are the frontline treatment for estrogen-dependent breast cancer. Our X-ray structural data from human placental aromatase (pArom) revealed an androgen-specific tight active site, unlike any other P450 [1]. Using a N-terminus transmembrane helix-truncated construct of human aromatase, we have now been able to crystallize the recombinant human enzyme (rArom), and confirm that the tertiary and active site structures of rArom are identical to those of pArom. The intermolecular interaction that dominates both crystals and probably drives their nucleation is via a surface loop roughly 39[Aring] away from the active site between helices D and E of one aromatase molecule that penetrates into the heme-proximal cavity of the next, forming a head to tail tandem aromatase array, akin to a polymer chain [2]. This oligomeric association between the negative potential surface of the D-E loop region and the positively charged heme-proximal cavity is rather unique among all known P450 structures. Oligomerization of aromatase on the lipid membrane has been previously reported [3,4]. We postulate that the observed head to tail association is a manifestation of the functionally and physiologically meaningful oligomeric state of aromatase. In order to investigate the effects of this head to tail association on aromatase function, we have constructed loop deletion and point mutants that disrupt the loop to proximal site interaction. In addition, two proximal site mutants that alter the surface electrostatic property have also been generated. All four D-E loop mutants generated thus far show a shift in the Soret Peak from high spin ferric to low spin ferrous, as well as greatly reduced enzyme activity and Soret peak heights. The results clearly demonstrate that the loop to proximal cavity intermolecular contact is required for the enzyme[apos]s catalytic function. Furthermore, the loop, being roughly 11A away from the neighboring heme iron, may even have a significant role in the stabilization of the heme-binding pocket of its neighboring aromatase molecule. Latest results on this continuing investigation will be presented.[br][br]1. Ghosh, D., et al., Structural basis for androgen specificity and oestrogen synthesis in human aromatase. Nature, 2009. 457(7226): p. 219-23. 2. Ghosh, D., et al., Higher order organization of human placental aromatase. Steroids, 2011. 76(8): p. 753-8. 3. Praporski, S., et al., Organization of cytochrome P450 enzymes involved in sex steroid synthesis: PROTEIN-PROTEIN INTERACTIONS IN LIPID MEMBRANES. J Biol Chem, 2009. 284(48): p. 33224-32. 4. Jiang, W., Ghosh, D., Motion and Flexibility in Human Cytochrome P450 Aromatase, PLoS One (in press).[br][br]Sources of Research Support: This research is funded in part by the grant R01GM086893 from the National Institutes of Health.[br][br]Nothing to Disclose: JL, GDN, JG, WJ, GG, DG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2163 68 628 SAT-539 PO25-01 Saturday 567 2012


568 ENDO12L_SAT-540 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Structure-Based Design of Novel Aromatase Inhibitors Debashis Ghosh, Jessica Lo, Daniel Morton, Susan Hubbell, Jingle Xi, Jing An, Chinaza Egbuta, Huw M Davies State University of New York (SUNY) Upstate Medical University, Syracuse, NY; Emory University, Atlanta, GA Human cytochrome P450 aromatase, an integral membrane hemeprotein of the endoplasmic reticulum, catalyzes with high specificity the synthesis of estrogens from androgens. Aromatase inhibitors (AIs) have thus become the frontline treatment for estrogen-dependent breast cancer. The crystal structure of the androstenedione-complex of the full-length aromatase purified from human placenta shows an exquisitely androgen-specific tight active site, unlike any other P450 structure [1]. New insights have been gained into the basis for substrate specificity, reaction mechanism, steroidal passage, vibrational modes, trans-membrane integration and oligomeric state [1,2]. The structural data have been utilized in rational design of novel aromatase-specific inhibitors based on the hypothesis that inhibitors that bind with high affinity at the aromatase active site, optimally utilize the available hydrophobic and polar interaction space and possess good shape complementarity with active site cleft, will be exclusive for human aromatase. We have thus far synthesized 78 new steroidal compounds and evaluated their inhibitory properties in highly purified placental aromatase with letrozole and exemestane, two currently used AIs, as the controls. A large number of these compounds exhibit strong inhibitory effects, and have IC50 values in the nM range, comparable to the AIs. The X-ray structures of aromatase complexes of some of these high potency inhibitors reveal their binding modes. In their enzyme-bound states, the novel side groups protrude into the only available space in the active site cleft, at the opening to the access channel [1], as the design considerations intended. Compounds with longer alkane and alkyne side groups that optimally fill the space have higher potency. The anti-proliferative effects of the most potent compounds are evaluated in a MCF-7 breast cancer cell line that stably expresses aromatase. Results on some of these compounds establish that most potent and exclusive inhibitors have EC50s of the order 1 nM, similar to AIs. Structure activity relationship obtained thus far on these compounds validate the concept that rational aromatase inhibitors designed based on the active site structure, property and binding mechanism would also have enhanced affinity and specificity for the target. Latest data on the continuing research for the development of aromatase-exclusive inhibitors will be presented.[br][br]1. Ghosh D, Griswold J, Erman M, Pangborn W. Structural Basis for Androgen Specificity and Oestrogen Synthesis in Human Aromatase. Nature 457: 219-223 (2009). 2. Jiang W, Ghosh D. Motion and Flexibility in Human Cytochrome P450 Aromatase. PLoS One (in press).[br][br]Sources of Research Support: This research is funded in part by the grant R01GM086893 from the National Institutes of Health.[br][br]Nothing to Disclose: DG, JL, DM, SH, JX, JA, CE, HMD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2168 68 629 SAT-540 PO25-01 Saturday 568 2012


569 ENDO12L_SAT-541 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Identification of Novel Inhibitors of 11beta-Hydroxysteroid Dehydrogenase Type 1 (11BHSD1) by [italic]In Silico[/italic] Screening and [italic]In Vitro[/italic] Confirmation Carlos F Lagos, Andrea Vecchiola, Juan Tichauer, Fidel Allende, Carolina Valdivia, Cristobal A Fuentes, Sandra Solari, Carmen Campino, Rene Baudrand, Mariana Cifuentes, Gareth I Owen, Cristian A Carvajal, Carlos E Fardella Pontificia Universidad Catolica de Chile, Santiago, Chile; Pontificia Universidad Catolica de Chile, Santiago, Chile; Pontificia Universidad Catolica de Chile, Santiago, Chile; Pontificia Universidad Catolica de Chile, Santiago, Chile; Universidad de Chile, Santiago, Chile; Millennium Institute of Immunology and Immunotherapy, Santiago, Chile The incidence of obesity and metabolic syndrome (MetS) continues to rise at an alarming rate. The enzyme 11BHSD1, regenerates cortisol (F) from inactive cortisone in liver and adipose tissue, and has been associated with central obesity, insulin resistance and hypertension. Several 11BHSD1 inhibitors are been developing for the treatment of diabetes and MetS. [bold]Aim.[/bold] To identify novel non-steroidal potential 11BHSD1 inhibitors using virtual screening techniques and in vitro assays in human adipocyte LS14 cells. [bold]Methods.[/bold] Human 11BHSD1-ligand complexes were retrieved from the PDB and structurally aligned. Ligands were clustered using ECFP fingerprints, and ligand-based pharmacophore models were generated with vROCS. The OpenNCI database ([sim]260,000 compounds) was retrieved and filtered by ADME/Tox constraints with FILTER, and the conformers generated by OMEGA. Virtual screening of the conformer database was performed with FRED. Compounds were obtained from the Developmental Therapeutic Program (NCI DTP), and the cytotoxicity assessed using the MTS assay. 11BHSD1 inhibition assays were performed in differentiated LS14 cells with18[beta]-glycyrrhetinic acid control inhibitor. Each compound was assayed in increasing concentrations from 0.3 to 10[micro]M in presence of 150 [micro]M of cortisone (E). Cortisol (F) was quantified by LC-MS/MS. [bold]Results.[/bold] A set of 18 protein-ligand complexes was structurally aligned. The RMSD for the superimposition of all proteins was less than 2[Aring]. Ligand clustering results in 5 main groups, one of them containing only steroidal molecules. For the other 4 groups, enrichment rate of the pharmacophore models was [gt]75%. ADME profiling renders 195,242 drug-like molecules, and a final library of 13,827,438 conformers. The top 1000 hits from docking were filtered with the pharmacophore models, and the best 200 hits binding mode analyzed. The final 39 selected compounds obtained from the NCI DTP did not affect cellular viability. Inhibition assays identified 4 hits compounds displaying enzyme inhibitory activities between 30-70% in the low micromolar range. [bold]Conclusion[/bold]. A combined ligand and structure-based virtual screening approach allow us to identified novel non-steroidal inhibitory compounds of human 11BHSD1with desirable physicochemical properties, which may serve as promising lead compounds for further development.[br][br]Sources of Research Support: FONDEF D08i1087, FONDECYT 1100356 [amp] IMII P09/016-F grants. The authors thank OpenEye Software for academic license of their products (FILTER, OMEGA, vROCS [amp] FRED), and the DTP/NCI for kindly providing the compounds screened in this study. CFL and CAC are CONICYT PhD fellows.[br][br]Nothing to Disclose: CFL, AV, JT, FA, CV, CAF, SS, CC, RB, MC, GIO, CAC, CEF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 226 68 630 SAT-541 PO25-01 Saturday 569 2012


570 ENDO12L_SAT-542 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Profiling Intact Steroid Sulfates in Biological Samples by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS-MS) Christina E Galuska, Michaela F Hartmann, Stefan A Wudy Justus Liebig University, Giessen, Germany For the initialization of a biological response via specific receptors, steroid hormones have to be in an unbound, free form. Circulating sulfated steroids [ndash] so far considered as biologically inactive metabolites intended for elimination - are not able to pass the cell membrane. However, the discovery of membrane bound uptake carriers for sulfated steroids, such as SOAT (sodium dependent organic anion transporter) and its colocalization with intracellular steroid sulfatases and sulfotransferases, points to a biological role of steroid sulfates. To further characterize their biological functions, a highly sensitive and specific analytical method was required. LC-MS-MS is the technique of choice permitting physicochemical analysis of the intact steroid conjugate with highest reliability thus avoiding interferences e.g. cross reactivity associated with immunoassays. Our workflow consisted of various sample purification procedures such as protein precipitation and solid phase extraction with the use of stable isotope labelled analogues as internal standards. After short liquid chromatographic separation the steroids were identified and quantified via specific mass transitions by electrospray triple quadrupole mass spectrometry in the negative ionization mode. We are currently able to simultaneously measure estrone sulfate, estradiol sulfate, 16-OH-dehydroepiandrosterone sulfate, androstenediol sulfate, dehydroepiandrosterone sulfate and pregnenolone sulfate in one profile. The method can be applied to various biological media such as plasma, cell lysates, methanolic extracts and was assessed for linearity, sensitivity, stability, recovery, matrix effects, as well as precision.[br][br]Sources of Research Support: Deutsche Forschungsgemeinschaft Research Group DFG FOR 1369 [ldquo]Sulfated Steroids in Reproduction[rdquo] (subproject 5).[br][br]Nothing to Disclose: CEG, MFH, SAW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1459 68 631 SAT-542 PO25-01 Saturday 570 2012


571 ENDO12L_SAT-543 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Dietary Sodium Modulates the Beta-2 Adrenergic Receptor-Mediated Mechanisms of Aldosterone Secretion Elijah T Trefts, Tham M Yao, Amanda E Garza, Alexander W Krug, Gail K Adler, Gordon H Williams, Luminita H Pojoga Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA [bold]Purpose :[/bold] The Beta-2 Adrenergic receptor ([beta][sub]2[/sub]AR) is a G-protein coupled receptor involved in blood pressure homeostasis. Recently, we found an association between human [beta][sub]2[/sub]AR genetic variants and salt sensitive hypertension, increased aldosterone (ALDO) but decreased plasma renin activity, suggesting excess ALDO production. However, it is unclear whether [beta][sub]2[/sub]AR is present in the adrenal cortex and whether [beta][sub]2[/sub]AR-specific stimulation induces ALDO secretion in a salt-dependent manner. The aim of this study was to address these questions in mechanistic studies performed on acutely isolated rat Zona Glomerulosa (ZG) cells.[br][bold]Methods :[/bold] Adult, male Wistar rats were maintained on a liberal salt (HS, 1.6% Na) or a low salt diet (LS, 0.02% Na). Adrenal cortex tissues were analyzed for [beta][sub]1[/sub]AR and [beta][sub]2[/sub]AR proteins. For ALDO secretion experiments, ZG cells were incubated with [beta][sub]2[/sub]AR-specific agonist (Procaterol, 10[sup]-4[/sup]-10[sup]-12[/sup] M). Samples were incubated in the presence/absence of 10[sup]-5[/sup] M [beta][sub]1[/sub]AR and [beta][sub]2[/sub]AR-specific antagonists (CGP 20712, ICI-118,551). ZG cells were also incubated with the same agonists in the presence/absence of Pertussis (PTX, 100 ng/ml) or Cholera Toxin (CTX, 150 ng/ml) or PLC inhibitor (U73122, 10[sup]-5[/sup] M). ALDO levels were measured in duplicate by radioimmunoassay. Data were reported as change relative to baseline (ng/10[sup]6[/sup] cells).[br][bold]Results :[/bold] Both [beta][sub]1[/sub]AR and [beta][sub]2[/sub]AR proteins were present in the rat adrenal cortex; however, only [beta][sub]2[/sub]AR levels were modulated by dietary salt (Higher on HS vs. LS). Procaterol induced significant (P[lt]0.001) ALDO secretion (LS: 551[plusmn] 126; HS: 47[plusmn] 23 ng/10[sup]6[/sup] cells) from ZG cells, with a calculated EC[sub]50[/sub] of 9.9 nM (HS) and 4.9 nM (LS). Stimulation was completely ablated by [beta][sub]2[/sub]AR-specific, but not by [beta][sub]1[/sub]AR-specific antagonism. G[sub]i[/sub] inhibition by PTX or G[sub]s[/sub] stimulation by CTX significantly increased the [beta][sub]2[/sub]AR-stimulated ALDO secretion on LS (control: 99.5[plusmn]7.86; PTX: 152.3[plusmn]15.33, P[lt]0.01; CTX: 304.8[plusmn]49.75, P[lt]0.001) but not on HS. U73122 inhibited the response partially on LS and completely on HS.[br][bold]Conclusions :[/bold] Our findings are consistent with the presence of a dietary sodium-sensitive [beta][sub]2[/sub]AR in the rat adrenal. Moreover, we identified two sodium-dependent adrenal mechanisms of [beta][sub]2[/sub]AR-stimulated ALDO secretion: a G[sub]i[/sub]/G[sub]s[/sub]/PLC-sensitive pathway on a LS diet, and a G[sub]i[/sub]/G[sub]s[/sub]-insensitive, PLC-sensitive pathway on a HS diet. Thus, these data support a potential role of polymorphic variants of the [beta][sub]2[/sub]AR gene in the adrenal, as mediators of salt sensitive hypertension in humans.[br][br]Nothing to Disclose: ETT, TMY, AEG, AWK, GKA, GHW, LHP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1032 68 632 SAT-543 PO25-01 Saturday 571 2012


572 ENDO12L_SAT-544 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Expression of P450c17 in the Human Fetal Nervous System Marcus D Schonemann, Marcus Muench, Meng-Kian Tee, Walter L Miller, Synthia H Mellon University of California, San Francisco, CA; University of California, San Francisco, CA; University of California, San Francisco, CA P450c17 catalyzes steroid 17[alpha]-hydroxylase and 17,20 lyase activities. P450c17 is expressed in human fetal and postnatal adrenals and gonads, and in the developing mouse nervous system, but little is known about its expression in the human nervous system. We obtained portions of 9, 10 and 11-week gestation human fetal peripheral nervous system and delineated the pattern of expression of P450c17 by immunocytochemistry using the P450c17 antiserum previously used to characterize P450c17 in the mouse brain. P450c17 was readily detected in the dorsal root ganglia (DRG) and spinal cord. Neural structures were identified with antisera to the cytoskeletal protein NCAM; DRGs were identified with antisera to the neuronal transcription factor BRN3A and neurotrophin receptor TrkB. The identification of P450c17 was confirmed using commercial antisera directed against different domains of P450c17 and by using antisera immunodepleted with authentic human P450c17. We also found expression of the cholesterol side-chain cleavage enzyme, P450scc, in the spinal cord and DRGs. Expression of P450scc is limited to the cell bodies of the DRGs; we never detected P450scc in fiber tracts from the ganglia. P450c17 expression was weak at 9 week[apos]s gestation; however, at this time, but not at 11 week[apos]s gestation, its expression was seen in circular structures that resembled lamellipodia, suggesting that P450c17 expression may define axonogenesis in the early embryonic DRG. In addition to expression in spinal cord and DRG, P450c17 was also identified in the trigeminal ganglia, previously shown in the embryonic mouse. Catalysis by P450c17 requires electron donation from P450 oxidoreductase (POR). Dual-label immunohistochemistry detected P450c17 and POR co-localized in DRG bundles, but fibers containing P450c17 lacked POR. These data suggest that neurosteroids synthesized via these two enzymes may act in the developing human nervous system. The expression of P450c17 in structures lacking POR means that P450c17 cannot be steroidogenic in those locations suggesting that P450c17 may have additional functions that do not require POR. These data demonstrate that like other species, P450c17 is expressed in the developing human nervous system. Newly identified functions of DHEA as a ligand for the neurotrophin receptor TrkA, further suggests that local DHEA synthesis may have neurotropic functions in the early human embryo.[br][br]Nothing to Disclose: MDS, MM, M-KT, WLM, SHM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1674 68 633 SAT-544 PO25-01 Saturday 572 2012


573 ENDO12L_SAT-545 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Ligands for Vitamin D Receptor (VDR) Elicit CYP3A Expression in Prostate Cancer Cells and Enhance the Metabolic Inactivation of Androgens Declan Doherty, Orla Maguire, Catherine Pollock, Paul D Thompson University of Ulster, Coleraine, UK; Roswell Park Cancer Institute, Buffalo, NY Prostate cancer (PCa) is the second most common cancer in men worldwide. Androgen deprivation therapy (ADT) which targets testicular androgen production is widely used for the treatment of locally advanced PCa. ADT is initially very effective however it almost always relapses to form castrate resistant prostate cancer (CRPC). In recent years a contributing factor towards the emergence of CRPC is the ability of the tumour to produce its own [apos]intracrine[apos] tumoural androgens from cholesterol and DHEA which promotes prostate cancer progression. Therefore effective treatment of high risk PCa may require the targeting of both circulating and tumoural androgens to halt continuing androgen signalling and pathogenesis.[br]We show here that ligands of the Vitamin D Receptor (VDR) can specifically induce expression of CYP3A genes in PCa cells. CYP3A genes can induce oxidative inactivation of a large number of substrates including androgens such as testosterone. Quantitative RT-PCR show CYP3A4 and CYP3A5 mRNA are significantly induced after treatment of LNCaP and LAPC-4 cells with vitamin D (1,25D3) and the vitamin D analogue seocalcitol (EB1089). In LNCaP cells CYP3A4 mRNA expression was induced 139-fold and 483-fold by 10nM 1,25D3 and EB1089, respectively. VDR regulation of these mRNA[apos]s was found to be time and dose-dependent. Using gene promoter assays and chromatin immunoprecipitation technology, we show that in PCa cells the CYP3A4 and CYP3A5 gene promoters are directly regulated by VDR via association of the receptor to both DR3 and ER6 type elements. VDR induced expression of the CYP3A4 protein resulting in a corresponding increase in CYP3A4 enzyme activity in LNCaP cells. Mass spectrometry analysis showed that treatment of LNCaP cells with VDR ligands resulted in an increased inactivation of testosterone to its 6-[beta]-hydroxy-testosterone metabolite (6-[beta]-OH-T). EB1089 was more potent than vitamin D at inducing CYP3A4 mRNA, protein and enzyme activity. The EC50 for CYP3A4 and CYP3A5 mRNA induction for EB1089 was much lower than that of 1,25D3 indicating its therapeutic potential.[br]This data highlights a potential application of VDR-based therapies as a means to reduce the bio-availability of growth-promoting androgens within the tumour micro-environment and limit the emergence of the castrate-resistant form of prostate cancer that often develops during the course of androgen deprivation-based therapies.[br][br]Sources of Research Support: Funded by the Prostate Cancer Charity, UK.[br][br]Nothing to Disclose: DD, OM, CP, PDT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 473 68 634 SAT-545 PO25-01 Saturday 573 2012


574 ENDO12L_SAT-546 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) HSD11B1 Promoter Binding of the NF-[kappa]B p65 Subunit in Response to TNF-[alpha] Suppresses 11[beta]-HSD1 Expression and Activity in C2C12 Muscle Cells Craig L Doig, Agnieszka E Zielinska, Jamila Bashir, Philip Guest, Paul M Stewart, Gareth G Lavery University of Birmingham, Birmingham, UK A rise in glucocorticoid (GC) levels is an important part of the innate immune response, that when impaired is associated with high mortality in inflammatory states. 11[beta]-hydroxysteroid dehydrogenase (11[beta]-HSD1) activity regenerates cortisol from cortisone (corticosterone from 11-DHC in rodents), acting at a pre-receptor level governing local GC concentrations and enhancing circulating levels. Pro-inflammatory cytokines such as TNF-[alpha] regulates the expression and activity of 11[beta]-HSD1 in some cell types, but the molecular mechanisms by which this occurs is unclear. In skeletal muscle, TNF-[alpha] is highly pleiotropic, it not only inhibits terminal differentiation of myoblasts, but also acts as a pro-mitogenic immune modulator enhancing satellite cell mediated muscle remodelling. C2C12 mouse muscle cells express 11[beta]-HSD1 and we hypothesised that its activity would be responsive to TNF-[alpha] through an NF-[kappa]B dependent mechanism. Differentiated C2C12 myotubes were challenged with TNF[alpha] (10ng/ml) for 24 hours and 11[beta]-HSD1 enzyme expression and activity examined. We showed that enzyme activity was reduced by approximately 30% compared to control (p[lt]0.01), and this was concomitant with a 3-fold reduction in mRNA and a similar reduction in 11[beta]-HSD1 protein when examined by western blot. We then used in-silico screening to identify 2 putative NF-[kappa]B p65 binding sites in the 11[beta]-HSD1 P1 and P2 promoters and used ChIP analysis to identify whether binding occurred under TNF-[alpha] stimulation in the differentiated C2C12 myotubes. We did not detect binding in the P2 promoter; however, a TNF-[alpha] dependent p65 translocation to the nucleus was detected and binding at position -156bp of the P1 promoter within 1 hour and was still detectable after 24h. These data suggest that acute TNF-[alpha] exposure in differentiated skeletal muscle activates the Nf-kB component p65 resulting in transcriptional down-regulation of 11[beta]-HSD1. These data suggest that acute TNF-[alpha] exposure causes a down-regulation of 11[beta]-HSD1 and therefore, the ability to locally augment the autocrine generation of GC. Functionally this response to TNF-[alpha] may serve to prolong the inflammatory response through decreasing local GC formation sufficiently to allow an appropriate response to occur. This mechanism may ultimately be detrimental in situations in which extended TNF-[alpha] exposure and chronic inflammation can lead to adverse outcomes and chronic muscle wasting disorders, which would otherwise respond well to GC treatment.[br][br]Nothing to Disclose: CLD, AEZ, JB, PG, PMS, GGL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2040 68 635 SAT-546 PO25-01 Saturday 574 2012


575 ENDO12L_SAT-547 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Glucocorticoid Regeneration in Skeletal Muscle Is Not Restricted to the Myocyte; Evaluation of the Skeletal Muscle-Specific 11[beta]-Hydroxysteroid Dehydrogenase Type 1 Knockout Mouse Agnieszka E Zielinska, Craig L Doig, Emma McCabe, Khalid M Saqib, Stuart A Morgan, Paul M Stewart, Gareth G Lavery University of Birmingham, Birmingham, UK The reactivation of glucocorticoids (GCs), cortisol to cortisone in humans and 11-DHDC to corticosterone in rodents, by the enzyme 11[beta]-hydroxysteroid dehydrogenase type 1 (11[beta]-HSD1), has been demonstrated in a number of metabolically important tissues such as liver and adipose. In muscle, GC excess promotes proteolysis, insulin resistance, and altered lipid homeostasis. 11[beta]-HSD1 inhibitor-treated and knockout (KO) mice resist diet induced obesity, retain insulin sensitivity and are athero-protected, but the full range of tissues mediating these [apos]cardio-protective[apos] phenotypes is not fully understood. Recently, we and others have demonstrated that 11[beta]-HSD1 expression regulates GC-induced insulin resistance and atrophy pathways in C2C12 muscle cells, mouse and human primary skeletal muscle cultures and may therefore be important in mediating muscle insulin sensitivity and sarcopenia. To further investigate 11[beta]-HSD1 and GC availability in skeletal muscle, we have generated muscle-specific KO mice (MKO) using Cre-LoxP technology, crossing a conditional HSD11B1 allele with Acta1-Cre transgenic mice, targeting 11[beta]-HSD1KO specifically to striated skeletal muscle. Initial characterisation of MKOs showed that recombination of the conditional to a KO allele was skeletal muscle-specific ([gt]90% of myocyte alleles recombined), and that all other tissues, including the heart were unaffected. However, muscle beds including quadriceps, tibialis anterior and soleus from MKOs assessed by real-time PCR for 11[beta]-HSD1 mRNA showed only a non significant, approximately 1.4 fold reduction (n=4) compared to control mice. Similarly, ex vivo muscle explants and skeletal muscle microsome preparations from MKO mice revealed no reduction of 11[beta]-HSD1 activity or expression by western blot compared to control mice. The Acta1-Cre is a well validated and extensively used transgenic and our 11[beta]-HSD1 liver-specific KO has complete loss of hepatic 11[beta]-HSD1 activity, and a similar degree of recombination efficiency as seen in MKO mice. We hypothesise that there are additional, non-myocyte cell types with high 11[beta]-HSD1 activity present within skeletal muscle (e.g. myofibroblasts) that may be important, and further analysis is warranted to ascertain their role in skeletal muscle function and metabolism.[br][br]Nothing to Disclose: AEZ, CLD, EM, KMS, SAM, PMS, GGL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1624 68 636 SAT-547 PO25-01 Saturday 575 2012


576 ENDO12L_SAT-548 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Short Pathway for Cortisol Catabolism and Stress Reduction in Zebrafish Janina Tokarz, Wiliam Norton, Gabriele Moeller, Martin Hrabe de Angelis, Jerzy Adamski Helmholtz Zentrum Muenchen, Neuherberg, Germany; Centre Nationale de la Recherche Scientifique, Gif sur Yvette, France Stress is a physiological reaction of an organism to a stressor, e.g. environmental challenges or predators. The stressor activates the hypothalamus-pituitary-adrenal gland axis, which leads to an increase of cortisol in circulation. Cortisol is the major stress hormone regulating glucose metabolism and suppression of the immune system, thereby mediating appropriate fight-or-flight reactions to the stressor.[br]Cortisol is efficiently inactivated by oxidation to cortisone by 11beta-hydroxysteroid dehydrogenase type 2. We have recently identified an enzyme putatively involved in further catabolic steps of cortisol inactivation in zebrafish (Danio rerio). This enzyme, 20beta-HSD type 2, catalyzes the conversion of cortisone to 20beta-hydroxycortisone (1). Treatment of zebrafish embryos with cortisol resulted in a strong up-regulation of both 11beta-HSD type 2 and 20beta-HSD type 2 transcripts in all developmental stages analyzed. Short-term swirling stress of 5 dpf zebrafish larvae showed a fast up-regulation of 11beta-HSD type 2 and 20beta-HSD type 2 as well as some more genes (e.g. crf and star) involved in the stress axis. Morpholino-induced 20beta-HSD type 2 knock-down fish larvae (morphants) showed no changes in development under normal conditions. However, upon challenging the morphants with cortisol, we observed a characteristic cortisol phenotype described earlier (2). This demonstrates a diminished capability of morphants to catabolize excess cortisol. In reporter gene experiments, we found that 20beta-hydroxycortisone is not a ligand for either the glucocorticoid or the mineralocorticoid receptor. The analysis of glucocorticoids in adult fish-holding water revealed 20beta-hydroxycortisone to be conjugated with glucuronic acid or sulfates, indicating its function as excretion product.[br]Taking these results together, we propose that the subsequent action of 11beta-HSD type 2 and 20beta-HSD type 2 as well as glucuronyl- and sulfotransferases represents a metabolic pathway leading to rapid deprivation of cortisol and its excretion as 20beta-hydroxycortisone. This short catabolic pathway of cortisol inactivation might therefore also be of importance in stress reduction.[br][br](1) Tokarz J, Mindnich R, Norton W, M[ouml]ller G, Hrab[eacute] de Angelis M, Adamksi J (2011): Discovery of a novel enzyme mediating glucocorticoid catabolism in fish: 20beta-Hydroxysteroid dehydrogenase type 2, MCE [Epub ahead of print]. (2) Hillegass JM, Villano CM, Cooper KR, White LA (2008): Glucocorticoids alter craniofacial development and increase expression and activity of matrix metalloproteases in developing zebrafish (Danio rerio). Toxicological Sciences 102, 413-424.[br][br]Nothing to Disclose: JT, WN, GM, MHdA, JA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 54 68 637 SAT-548 PO25-01 Saturday 576 2012


577 ENDO12L_SAT-549 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Anti-Inflammatory Effect of a High Dose of Corticosteroids Is Associated with Some Paradoxical Pro-Inflammatory Effects Paresh Dandona, Husam Ghanim, Sandeep Dhindsa, Kelly Green, Sanaa Abuaysheh, Katherine Frachetti State University of New York at Buffalo, Buffalo, NY We have previously demonstrated that a low dose of hydrocortisone (100 mg) given intravenously suppresses intranuclear NF[kappa]B and AP-1 binding and the expression of pro-inflammatory genes like MMPs. We have now investigated the effect of a high dose of hydrocortisone (300mg=60 mg prednisolone) on NF[kappa]B binding and the expression of TLRs, the mediators of TLR signal transduction, MyD88 and TRIF and HMG-B1. Ten normal subjects were injected intravenously with 300mg of hydrocortisone or saline in 2 separate visits one week apart in a randomized crossover study. Blood samples were obtained at 0,2,4,8 and 24h after the injection. Mononuclear cells (MNC) were prepared by standard techniques and were tested for NF[kappa]B binding and the expression of TLRs, MyD88, TRIF, chemokines and chemokine receptors and HMG-B1. Plasma concentrations of glucose, FFAs, NO metabolites, chemokines and HMG-B1 were also measured. Following the injection of this dose, there was a significant increase in glucose concentration from 92[plusmn]4 to 116[plusmn]6 mg/dl, a marked increase in FFA concentrations from 0.38[plusmn]0.1 to 0.804[plusmn]0.15mM. While NF[kappa]B binding and the mRNA expression of MyD88, TRIF, chemokines and chemokine receptors were suppressed significantly in MNC, the mRNA expression of TLR 2, 5 and 9 and HMG-B1 was increased (by 103[plusmn]24%, 107[plusmn]19%, 56[plusmn]13% above the baseline, respectively) in the MNC as was the concentration of HMGB1 (by 37[plusmn]12%) and MMP-9 (125[plusmn]22%) in plasma. Thus, while this high dose of HC exerts a powerful anti-inflammatory effect as shown above, it also exerts certain paradoxical pro-inflammatory effects. Since both glucose and FFAs have been shown to be pro-inflammatory, it is possible that they contribute to these effects. These paradoxical pro-inflammatory effects may account for the inability of these drugs to show benefit in clinical trials of septicemia and other severe pro-inflammatory states.[br][br]Disclosures: PD: Speaker, Novo Nordisk. Nothing to Disclose: HG, SD, KG, SA, KF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2146 68 638 SAT-549 PO25-01 Saturday 577 2012


578 ENDO12L_SAT-550 POSTER SESSION: Steroid Hormone Biosynthesis [amp] Metabolism (1:30 PM-3:30 PM) Evaluation of the Potential for Inhibition of Steroidogenesis by an Oncology Drug in the Human H295R Adrenocortical Cell Line and Rat Adrenocortical Cells Katie Kubek, Aaron Nelson, Salah-Dine Chibout, Francois Pognan, Nancy Turner, Patrick John Devine Novartis Institutes for Biomedical Research Inc, Cambridge, MA; Novartis Institutes for Biomedical Research Inc, Emeryville, CA Compound ABC123 (a new molecular entity, identity not disclosed), being developed for an oncology indication, induced vacuolation and hypertrophy in the zona fasciculata of adrenal glands in both rats and dogs following repeated daily oral dosing. [italic]In vitro[/italic] and [italic]in vivo[/italic] studies were initiated to investigate the mechanism by which ABC123 impacts adrenal function. Potential ABC123-induced cytotoxicity or alterations in forskolin- and ACTH-stimulated production of progesterone (prog), cortisol/corticosterone (cort), and aldosterone (aldo) were assessed in a human adrenocortical cell line, H295R cells, and in primary rat adrenocortical cells. A known inhibitor of steroidogenesis, etomidate, was used as a positive control. Also, adrenal histology and prog, cort, aldo, testosterone, and ACTH levels were measured in rats following a single dose or 7 days of oral dosing with 7.5 or 75mg/kg/day ABC123. Viability in both H295R cells and primary adrenocortical cultures was not affected by etomidate and was significantly reduced with [ge]33[micro]M ABC123. In H295R cells, etomidate significantly inhibited prog, cort and aldo production at all concentrations tested (0.14-100[micro]M). ABC123 significantly reduced synthesis of all three steroids at [ge]0.4[micro]M. With forskolin-stimulated rat adrenocortical cells, etomidate caused inhibition of aldo, cort and prog production at [ge]4[micro]M, [ge]0.4[micro]M and [ge]11[micro]M, respectively, whereas inhibition occurred at [ge]0.1[micro]M for aldo and [ge]4[micro]M for cort and prog in ACTH-stimulated primary cells. In contrast, ABC123 caused significant reductions in aldo, cort and prog at or above 4[micro]M in stimulated primary cells. [italic]In vivo[/italic], ABC123 did not induce any significant differences in the tested steroids among treatment groups, but ACTH was elevated and vacuolation of the adrenal cortex was observed with 75mg/kg for 7 days. ACTH increase without a concomitant increase in cort has been reported with compounds that inhibit adrenal steroidogenesis. Both the reduced steroid production [italic]in vitro[/italic] and increased ACTH [italic]in vivo[/italic] may suggest such a mechanism by which ABC123 alters adrenal histology. Inhibition of steroid biosynthesis by ABC123 and etomidate was demonstrated [italic]in vitro[/italic], but further investigations with precursor steroids in cultures would be needed to examine what step(s) in steroidogenesis is/are affected by ABC123.[br][br]Disclosures: KK: Employee, Novartis Pharmaceuticals. AN: Employee, Novartis Pharmaceuticals. S-DC: Employee, Novartis Pharmaceuticals. FP: Employee, Novartis Pharmaceuticals. NT: Employee, Novartis Pharmaceuticals. PJD: Employee, Novartis Pharmaceuticals. 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1278 68 639 SAT-550 PO25-01 Saturday 578 2012


579 ENDO12L_SAT-559 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Effects of Soy as an Endocrine-Disrupting Chemical on Granulosa Cell Tumor Progression Nadera Mansouri-Attia, Rebecca James, Stephanie A Pangas Baylor College of Medicine, Houston, TX The potential effect of endocrine disrupting chemicals (EDCs) such as phytoestrogens is becoming increasingly evident on both human and animal health. The effects on the reproductive tract and the developing embryo are of a particular concern, as are the role of EDCs in the initiation and progression of diseases including cancer. Isoflavones are a category of phytoestrogens found in soy with both estrogenic and anti-estrogenic effects whose role in tumors initiation, progression or suppression has yielded conflicting results. However, data from rodent studies suggest that neonatal exposure to genistein (the predominant isoflavone in soy) disrupts the normal reproductive function even though its role in ovarian cancer and particularly in granulosa cell tumors (a rare form of human ovarian cancer) is largely unkown. To decipher the molecular mechanisms leading to the initiation and the progression of granulosa cells tumors (GCT) and to determine the potential effect of isoflavones in tumoral spread, we used a female conditional knockout mice model for the BMP signaling SMADs (Smad1/5 dKO) in the ovary which are infertile as adults and develop metastatic GCT. We compared Smad1/5 dKO and control mice maintained on a non-soy or a soy-based diet post weaning for one year. Preliminary results demonstrate that Smad1/5 dKO mice fed a non-soy based diet showed similar fertility defects that were found previously for those fed a soy-based diet. However, only 50% of the mice on a non-soy diet developed GCT during the one-year time frame. Furthermore, switching mice bred and raised on a non-soy vegetarian diet to a soy-based diet rapidly increased tumor incidence 60% by two months, and 100% by three. Interestingly, Smad1/5 dKO tumors showed changes in expression of the estrogen receptor alpha isoform (ER[alpha]) even though ER[beta] is considered the predominant ER isoform in ovarian development. We hypothesize that genetic background may determine the effect of soy on cancer growth, and that the BMP SMADs play an important and currently uncharacterized role in modulating estrogen signaling, possibly through differential regulation of the estrogen receptors and which ultimately affects growth and metastasis of tumors cells.[br][br]Sources of Research Support: NIH Grant CA138628 and a Burroughs Wellcome Career Award in the Biomedical Sciences to S. Pangas.[br][br]Nothing to Disclose: NM-A, RJ, SAP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1957 69 640 SAT-559 PO24-01 Saturday 579 2012


580 ENDO12L_SAT-560 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Neonatal Phytoestrogen Exposure Alters Oviduct Mucosal Immune Response to Pregnancy and Affects Preimplantation Embryo Development in the Mouse Wendy N Jefferson, Elizabeth Padilla-Banks, Amy M Cantor, Carmen J Williams National Institute of Environmental Health Sciences, Research Triangle Park, NC Treatment of neonatal mice with the phytoestrogen genistein (50 mg/kg/day) results in complete female infertility that is caused in part by preimplantation embryo loss in the oviduct between days 2 and 3 of pregnancy. We previously demonstrated that oviducts of genistein-treated mice are [ldquo]posteriorized[rdquo] as compared to control mouse oviducts because they express numerous genes normally restricted to posterior regions of the female reproductive tract (FRT), the cervix and vagina. We report here that neonatal genistein treatment resulted in substantial changes in oviduct expression of genes important for the FRT mucosal immune response, including immunoglobulins, antimicrobials, and chemokines. Some of these altered immune response genes were chronically altered beginning at the time of neonatal genistein treatment, indicating that they were a result of the posteriorization phenotype. Other alterations in oviduct gene expression were observed only in early pregnancy, immediately after the FRT is exposed to inflammatory or antigenic stimuli from ovulation and mating. The oviduct changes affected development of the surviving embryos by increasing the rate of cleavage and decreasing the ratio of trophectoderm to inner cell mass cells at the blastocyst stage. We conclude that both altered immune responses to pregnancy and deficits in oviduct support for preimplantation embryo development in the neonatal genistein model are likely to contribute to the infertility phenotype.[br][br]Nothing to Disclose: WNJ, EP-B, AMC, CJW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1951 69 641 SAT-560 PO24-01 Saturday 580 2012


581 ENDO12L_SAT-561 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Non-Genomic Estrogenic Effects of the Phytoestrogen Metabolite R-Equol and the Pharmaceutical Birth Control Estrogen Ethinyl Estradiol in GH3/B6/F10 Pituitary Tumor Cells Manish Kumar Saraf, Cheryl S Watson University of Texas Medical Branch, Galveston, TX Estradiol (E[sub]2[/sub]), a physiological estrogen, activates various non-genomic signaling pathways via plasma membrane estrogen receptors (mERs). Ethinyl estradiol (EE[sub]2[/sub]) is an orally bio-active estrogen used in formulations of contraceptives. The actions of both endogenous estrogens and therapeutic pharmaceutical estrogens are likely to be influenced by estrogen mimics found in food sources. The phytoestrogen equol is a metabolite of the soy isoflavone daidzein that is influenced by gut flora. We have previously reported that some phytoestrogens and environmental estrogens also act as non-genomic phospho-activators of various mitogen-activated protein kinases (MAPKs) in mER[alpha]-enriched GH3/B6/F10 rat pituitary tumor cells. In the present study we extended these studies to examine R-equol- and EE[sub]2[/sub]-induced ERK, JNK and p38 phosphorylation, quantified at different time points and concentrations, using a plate immunoassay. E[sub]2[/sub], EE[sub]2[/sub],[sub] [/sub]and R-equol all induced pERK activity rapidly, with initial peaks in the 2.5-10 min range, with subsequent oscillations as we have observed for other estrogens previously. EE[sub]2 [/sub]and R-equol were more efficacious than E[sub]2[/sub] at expected blood level concentrations. All three estrogens rapidly induced p38 activity, though the equol-induced activation peak was delayed to 15 min. Similarly all three estrogens rapidly activated JNK, with a delayed peak activation (to 10 min) by R-equol and EE[sub]2[/sub], but a more robust activation by E[sub]2[/sub]. We can conclude that R-equol and EE[sub]2 [/sub]are rapid and potent activators of the MAPKs ERK, JNK and p38, similar to their activation by E[sub]2[/sub], but with slight differences in timing patterns and efficacies. Such actions suggest explanations for the diverse biological actions of these molecules, raising questions about their combined actions, as they are often present together when contraceptives and foods containing these compounds are consumed.[br][br]Nothing to Disclose: MKS, CSW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2036 69 642 SAT-561 PO24-01 Saturday 581 2012


582 ENDO12L_SAT-562 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Isoflavone Exposure during Different Periods of Life Differentially Modulates Estrogenic Response of the Mammary Gland and the Fat Tissue Almut Molzberger, Tina Blei, Clarissa Gerhauser, Sabine Kulling, Leane Lehmann, Dennis Muller, Guenter Vollmer, Patrick Diel German Sports University, Cologne, Germany; Technical University Dresden, Dresden, Germany; University W[uuml]rzburg, W[uuml]rzburg, Germany; Max Rubner Institute for Nutrition, Karlsruhe, Germany; German Cancer Research Center, Heidelberg, Germany The incidence of breast cancer in eastern Asia is approximately 3-times lower than in western countries. There is evidence that the traditional East Asian diet which is rich Isoflavones (ISO) like genistein (GEN), daidzein (DAI), and glycitein (GLY) seems to play an important role and that ISO intake must be during certain windows of development to exert anti-cancerogenic action. In this study we investigate the effects of soy ISO exposure in different periods of life on the estrogen sensitivity of the mammary gland in four defined groups of a rat animal model.[br]Group 1 received lifelong an ISO-free diet (IDD), Group 2 lifelong a ISO-rich diet (ISD). Group3 was exposed during puberty to an ISD from PND 30 up to PND 60; (pISD). In Group 4 adult animals were exposed to a diet enriched with an ISO extract (IRD 400) starting in the age of 80 days. In IDD, ISD and pISD onset of puberty, the menstrual cycle length andproliferative status of the mammary gland at day 50 were determined. The onset of puberty occurred significant earlier in pISD and ISD compared to IDD. Menstrual cycle length was significantly shortened in pISD. In intact animals at day 50 and day 80 the uterus wet weights were not affected by the respective diets. Proliferation in the mammary gland was not affected by IDD and ISD.[br]To analyze the effect of ISO on the estrogen sensitivity of mammary gland, animals of all groups were ovariectomised at the age of 80 days. At the age of 94 days animals were treated for 3 days with estradiol. Interestingly proliferative response of the mammary gland towards E2 was significantly reduced in the pISD and ISD groups. In contrast a reduced response of progesterone receptor (PR) expression towards E2 could be only observed in the ISD group. In the IRD 400 group visceral fat tissue mass was significantly reduced by E2 treatment compared to the respective control group.[br]In summary our results provide evidence that in female rats the exposure to ISO during puberty reduces the proliferative response to E2. In contrast regulation of PR expression by E2 is only affected following lifelong ISO exposure. Our preliminary data from fat tissue suggest that in addition other tissues than the mammary gland differentially responds to estrogens following dietary exposure to ISO. We hypothesize that the estrogen sensitivity of different endpoints relevant to critical periods of development are modulated by ISO. Investigation of the involved molecular mechanism is in progress.[br][br]Sources of Research Support: Deutsche Forschungsgemeinschaft DFG.[br][br]Nothing to Disclose: AM, TB, CG, SK, LL, DM, GV, PD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 747 69 643 SAT-562 PO24-01 Saturday 582 2012


583 ENDO12L_SAT-563 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Resveratrol-Induced Tensin Expression in Prostate Cancer Cells Is Androgen Receptor-Dependent Courtney Pisano, Nisha Patel, Bingchan Zhang, Dianzheng Zhang Philadelphia College of Osteopathic Medicine, Philidelphia, PA; Thomas Jefferson University, Philadelphia, PA The chemopreventive effects of resveratrol (RSV) on cancers, including prostate cancer, have been well documented; however, the mechanisms are still elusive. It has been reported recently that tensin, a matrix-adhesion protein, which is greatly down-regulated in prostate cancer, has been induced by RSV in several cancer cell lines. We are interested in finding out if the regulatory effect of RSV on the expression of tensin is part of the mechanism of RSV[apos]s inhibition of prostate cancer development. We first treated the cell line derived from prostate cancer cell line LNCaP with RSV and demonstrated that tensin mRNA levels were upregulated in a time- and dose-dependent manner. Since LNCaP cells are androgen receptor (AR) positive and previous findings have shown that RSV not only down-regulates AR protein levels but also its transcriptional activity, we were interested to see whether RSV effects on tensin are AR-dependent. When LNCaP cells were treated with R1881, a synthetic androgen, tensin mRNA levels were down-regulated and this effect was counteracted by RSV. These results strongly indicate that RSV up-regulates tensin through the function of AR. To further confirm its AR dependency, we used a previously established AR(+) cells in a series of experiments, with the AR(-) cells serving as a negative control. Similar to that in the LNCaP cells, tensin mRNA levels were only up-regulated in the AR(+) cells, not in the AR(-) cells. Finally, we demonstrated that tensin protein levels are also up-regulated by RSV in LNCaP and AR(+) cells. However, this effect was not shown in the AR(-) cells. In summary, we have demonstrated that RSV up-regulates tensin expression in prostate cancer cells and this effect is, at least in part, AR-dependent. Together with our previous observations, this provides further evidence at both cellular and molecular levels that RSV could serve as a preventative and therapeutic agent for prostate cancer.[br][br]Nothing to Disclose: CP, NP, BZ, DZ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1592 69 644 SAT-563 PO24-01 Saturday 583 2012


584 ENDO12L_SAT-564 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Novel [italic]In Vitro[/italic] Estrogen Activity of Blueberry Extract Chioma Obih, Felicia Gibson, Candace Hopgood, Patience Obih, Thomas Wiese Xavier University of Louisiana, New Orleans, LA This project tests the hypothesis that blueberry extract will induce the same level of estrogen activity in three in vitro bioassays. Blueberry extract (methanolic) was used to treat MCF-7 cells in a 7 day proliferation assay as well the MVLN and T47DkbLuc three day reporter gene assays. The blueberry extract induced an agonist response in the MCF-7 proliferation assay that could be inhibited with the antiestrogen ICI 182,780. At the same time, the blueberry extract induced only minimal agonist activity in the reporter gene assays (MVLV and T47DkbLuc). Of interest is the observation that the blueberry extract was able to block the activity of estradiol in both reporter gene assays, acting as a complete antagonist. This extract was also shown to bind the estrogen receptor alpha in a recombinant receptor FP assay. Thus, we report that the blueberry extract does not induce the same estrogen activity in three different in vitro bio assays. This extract can act as an estrogen agonist or antagonist depending on the cell and assay type. These studies highlight the potential for the blueberry extract to induce various and opposite estrogen activities when used as a dietary supplement.[br][br]Nothing to Disclose: CO, FG, CH, PO, TW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2310 69 645 SAT-564 PO24-01 Saturday 584 2012


585 ENDO12L_SAT-565 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) [italic]In Vivo[/italic] Anti-Androgenic Activity of Naringenin Type Phytoestrogens Oliver Zierau, Tina Blei, Georg Kretzschmar, Frank Josef Moeller, Guenter Vollmer Technische Universit[auml]t Dresden, Dresden, Germany Naringenin-type natural compounds are known for their estrogenic activity, but also showed anti-androgenic properties in two independent (anti-)androgen in vitro screening assays. Therefore we aimed to verify potentially their anti-androgenic activities in vivo by the comparative assessment of the activities of naringenin, 6-(1,1-dimethylallyl)naringenin (6-DMAN) and 8-prenylnaringenin (8-PN). Methodologically we performed an (Anti-)Hershberger assay in orchiectomized animals. Control animals were carrier treated, all other animals were substituted with dihydrotestosterone (0,25 mg/kg KGW/d) alone or in combination with the antiandrogens flutamide or bicalutamide (3 mg/kg KGW/d), the three phytoestrogens (15 mg/kg KGW/d) or estradiol (10 [micro]g/kg KGW/d). After seven day treatment the wet weight of the seminal vesicles and prostate was estimated. As expected, dihydrotestosterone increased accessory sex organ wet weights statistically. Flutamide and or bicalutamide significantly inhibited the dihydrotestosterone induced re-growth of seminal vesicles and ventral prostate. The phytoestrogens and estradiol also inhibited the re-growth of ventral prostate, but not to the same extent as the antiandrogens. In contrast the re-growth of seminal vesicles was only inhibited by 6-DMAN, but not by 8-PN, naringenin or estradiol. To further elucidate the potential antiandrogenic effects on a mechanistic level, we assessed expression patterns of selected genes in the accessory sex organs on the mRNA and protein level. In seminal vesicles and ventral prostate we analysed the mRNA levels of the estrogen receptors [alpha] and [beta], androgen receptor, clusterin and ornithin decarboxylase 1. Protein expression levels were assessed for prostate specific antigen, proliferating cell nuclear antigen and the androgen receptor. The molecular studies to large extend confirmed physiological findings on antiandrogenic properties of the tested naringenins. In summary, for the first time we provide evidence of the anti-androgenic activity of naringenin type phytoestrogens in vivo on the level of accessory sex organ weight, gene and protein expression. In conclusion, we gave further proof to the ability of prenylated naringenins not only to act via the estrogen receptor but also through the androgen receptor.[br][br]Nothing to Disclose: OZ, TB, GK, FJM, GV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1188 69 646 SAT-565 PO24-01 Saturday 585 2012


586 ENDO12L_SAT-566 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) In Utero Exposure of High-Fat Diet and Bisphenol A Reprograms Breast Cancer Risk Yuet-Kin Leung, Vinothini Janakiram, Robin Gear, Scott Belcher, Shuk-Mei Ho University of Cincinnati, Cincinnati, OH; University of Cincinnati, Cincinnati, OH Background: Exposure to environmental endocrine disruptors such as bisphenol A (BPA) has been shown to increase breast cancer risk and alter the window of cancer susceptibility. A number of studies have also suggested that lifestyle, especially adopting high fat diet, could predispose to higher breast cancer (BCa) risk. Though BPA and high-fat diet exposure have been independently investigated, the synergistic action of both factors in breast cancer has not been determined.[br]Objective: The objective of this study is to determine the in utero exposure of different oral doses of BPA in a high-fat diet background on BCa risk in late-life.[br]Approaches and results: The cancer risk was evaluated using 50 day-DMBA induced mammary tumorigenesis Sprague-Dawley rat model. BPA exposure in utero did not yield significant change in the pup[apos]s gender ratio, the food consumption and body weights of dams and PND1-42 day old female offspring compared with the high fat butter only or control diets. There was no significant difference in time of puberty onset in experimental and control daughters as determined by vagina opening. Forty days after single dose of DMBA treatment, the percentage of tumor incidence was increased in low dose BPA-treated animals that is, 62.5% in 2.5 [micro]g/kg BW and 100% in 25 [micro]g/kg BW BPA doses compared with higher doses (50% in 250 [micro]g/kg BW and 50% in 2500 [micro]g/kg BW) and control (55.55% in high fat only group) respectively, although no significant difference in tumor latency and multiplicity was observed.[br]Conclusion: In this study, we found that in utero exposure of BPA at 2.5 [micro]g/kg BW, in the presence of high butter diet background, showed significant effect in reprogramming the breast cancer risk using 50 day-DMBA mammary tumor model. Our data suggest that high fat diet may interact with BPA by advancing the windows of cancer susceptibility to DMBA.[br][br]Sources of Research Support: Acknowledgements: This work was support by National Institutes of Health grants NIEHS- U01ES019480 (YKL, SB, SMH) and P30ES006096 (SMH).[br][br]Nothing to Disclose: Y-KL, VJ, RG, SB, S-MH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2341 69 647 SAT-566 PO24-01 Saturday 586 2012


587 ENDO12L_SAT-567 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Fetal Exposures to Environmental Endocrine Disruptors Cause Long-Term Molecular Reprogramming of the Hypothalamus Deena M Walker, Ross Gillette, Andrea C Gore University of Texas at Austin, Austin, TX; University of Texas at Austin, Austin, TX; University of Texas at Austin, Austin, TX Polychlorinated Biphenyls (PCBs) are industrial contaminants and a class of endocrine disrupting chemicals. Exposure to PCBs during perinatal development results in long-term alterations in numerous reproductive endpoints in rodents such as the timing of puberty and reproductive senescence. However, little is known regarding the process by which these long-term alterations occur. We tested the hypothesis that early life exposure to Aroclor 1221 (A1221), a commercially available mixture of PCBs, results in long-term alterations to reproductive parameters and are associated with changes in gene expression in the hypothalamus. Methods: Pregnant Sprague Dawley rats were injected on embryonic day 16 and 18 with vehicle (DMSO; N=22) or A1221 (1mg/kg; N=23). Dams gave birth, and developmental milestones such as the timing of puberty and estrous cyclicity were monitored in the developing offspring. On postnatal days (P) 15, 30, 45, 90 and [sim]270, 1 male and 1 female from each litter was euthanized. Two brain regions important for reproductive function were punched: anteroventral periventricular nucleus (AVPV) and the arcuate nucleus (ARC) were collected and frozen for assessment of mRNA expression of a panel of 48 neuroendocrine genes. Results: There were developmental effects of prenatal EDCs, manifested as age-dependent alterations of gene expression, with effects most profound on P15 (males) and P45 (females). In females, A1221 exposure also resulted in altered estrous cyclicity in adulthood and in males the timing of puberty was delayed. Specifically, in the AVPV, 3 genes were affected in an age- and sex-specific manner by treatment with A1221: Kiss1, Kiss1r and Gper. In the ARC, Kiss1, Tac2 and Pdyn were all affected by treatment. Conclusions: These data suggest that gestational exposure to A1221 has long-term effects on genes in the AVPV and ARC that are important for GnRH secretion thereby providing a potential mechanisms for the altered phenotypes observed in adulthood. Additionally, the effects in the ARC and AVPV are age-, sex- and developmental stage-specific, suggesting that EDCs may alter development of the hypothalamus rather than just altering individual genes. Finally, these data provide evidence that the Kiss1/Neurokinin B/Dynorphin (KNDy) neurons in the ARC are potential targets of endocrine disruption. We are currently investigating if changes in DNA methylation are associated with these changes in gene expression observed in the hypothalamus.[br][br]Sources of Research Support: NIH F31 AG034813-01A1F31 [ndash] DMW. NIH RC1 ES018139 - ACG.[br][br]Nothing to Disclose: DMW, RG, ACG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 959 69 648 SAT-567 PO24-01 Saturday 587 2012


588 ENDO12L_SAT-568 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Chemical Mixtures of Bisphenol-A and Bisphenol-S Disrupt E2-Induced Non-Genomic Signaling in mER[alpha]-Enriched (GH3/B6/F10) Rat Pituitary Cell Line Rene Vinas, Cheryl S Watson University of Texas Medical Branch, Galveston, TX Bisphenol monomers are polymerized to form plastics used ubiquitously for food and water containers, and other consumer products. Stringent government regulations on the production and use of bisphenol-A (BPA) has led to the development of alternative, more heat stable bisphenol compounds as substitutes; one such alternative is bisphenol-S (BPS). We have previously shown rapid activation of several kinases by very low concentrations (fM-nM) of BPA and related alkylphenol compounds (eg. nonylphenol) in the GH3/B6/F10 rat pituitary cell line, acting via the membrane estrogen receptor (mER[alpha]). Because BPS may soon be introduced into the environment, this study aimed to determine if BPS is estrogenic, and if combining BPS and BPA with a physiological estrogen, estradiol (E2) affects signaling pathways. We measured the effects of these compounds both alone and together with E2, by quantifying E2-induced phosphorylations of extracellular signal-regulated kinases (ERKs) and c-Jun-N-terminal kinases (JNKs) in a high-throughput plate immuno-assay. BPS and BPA as lone compounds were both able to elicit rapid phospho-activation of ERK in a non-monotonic dose (fM-nM) and oscillating time-dependent (2.5-60min) manner. When BPA and BPS were combined with nM E2 the physiological estrogen[apos]s response was attenuated. Individual bisphenol compounds were unable to activate JNK; however, their combination with E2 generated an enhanced, rapid, and non-monotonic dose-response. The effects on these signaling pathways suggested that these bisphenol compounds may influence and perhaps promote cell proliferation. E2 and each bisphenol compound induced cell proliferation, while the bisphenol mixture with E2 suppressed proliferation below that of the vehicle control, suggesting a possible apoptotic response. Extrinsic caspase 8 activity was suppressed by E2, elevated by both bisphenol compounds at some concentrations, and unaffected by mixtures. Intrinsic caspase 9 activity was also inhibited by E2, and by some combinations (E2 + BPS + BPA [plusmn] nonylphenol) of xenoestrogens. Because E2 and both bisphenol compounds can alter apoptosis via these two pathways, they can contribute to changes in cell number. In mixtures expected to be found in contaminated environments, these compounds can have more dramatic disrupting effects.[br][br]Sources of Research Support: The Passport Foundation; NIEHS Grant 1F31ESO21164-01.[br][br]Nothing to Disclose: RV, CSW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2197 69 649 SAT-568 PO24-01 Saturday 588 2012


589 ENDO12L_SAT-569 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Neonatal Exposure to Bisphenol A Alters Steroid Hormones and GNRH-I Content in the Ovary of Adult Rats Nadia S Bourguignon, Maria M Bonaventura, Victoria A Lux-Lantos, Carlos Libertun Instituto de Biolog[iacute]a y Medicina Experimental-CONICET, Buenos Aires, Argentina; Universidad de Buenos Aires, Buenos Aires, Argentina; Universidad de Buenos Aires, Buenos Aires, Argentina Bisphenol A (BPA), is an endocrine disruptor with multiple effects; it behaves as an estrogen agonist and/or antagonist and as an androgen antagonist. In previous works we described the effects of BPA on the hypothalamic-pituitary-gonadal axis in female rats; we found that neonatal exposure to BPA was associated in adulthood with high serum estradiol (E2) and testosterone (T) levels, reduced serum progesterone (P), and altered in vitro hypothalamic GnRH secretion. Animals exposed to BPA also had altered ovarian morphology (1, 2, 3). Following this line, here we study the ovarian content of E2, P4, T and GnRH-I.[br]Female Sprague-Dawley rats were injected subcutaneously, daily from the day of birth to postnatal day (PND) 10, with BPA in castor oil at 50 [mu]g/50[mu]l (BPA50), 500 [mu]g/50[mu]l (BPA500), or castor oil (Control), as described in (1). Rats were sacrificed in the morning of estrus at 110-120 days of age. The ovarian content of E2, P, T and GnRH-I were evaluated by RIA. Results were expressed as means SE. Data were analyzed by one-way analysis of variance and were considered significant when p[lt]0.05.[br]We found higher ovarian E2 content (pg/mg ovary: C: 2.11[plusmn]0.55, BPA50: 12.00[plusmn]4.28 (p[lt]0.005), BPA500: 29.76[plusmn]7.85 (p[lt]0.0005), n= 8-14) and lower P4 (pg/mg ovary: C: 531.28[plusmn]29.57, BPA50: 351.59[plusmn]54.81 (p[lt]0.05), BPA500: 263.44[plusmn]56.84 (p[lt]0.005), n= 7-12). T content was not significantly different among groups (pg/mg ovary: C: 17.54[plusmn]4.57, BPA50: 11.13[plusmn]3.09, BPA500: 8.47[plusmn]1.72, n= 6-13). In BPA500, GnRH-I content was increased (pg/mg ovary C: 0.54[plusmn]0.05, BPA50: 0.69[plusmn]0.07, BPA500: 0.97[plusmn]0.07; BPA500: p[lt]0.0005 vs. C, p[lt]0.05 vs. BPA50, n= 9-11).[br]We conclude that in adult neonatally-BPA treated rats ovarian E2 content, increased, and P4 content, decreased, are altered coinciding with the serum levels previously reported. Ovarian T content did not change, unlike serum levels, suggesting changes in metabolism and/or origin of this steroid. In addition, BPA-treatment increased the ovarian GnRH-I content.[br]These results suggest that neonatal exposure to BPA produces a clear alteration in ovarian GnRH-I and steroidogenesis in adulthood.[br][br]1. Fernandez M et al. Environ Health Perspect 2009; 117:757. 2. Fernandez M et al. Environ Health Perspect 2010; 118:1217. 3. Bourguignon NS et al. The Endocrine Society[apos]s 92nd Annual Meeting. S Diego, CA, 2010. Abstract book. P2-59.[br][br]Sources of Research Support: Agencia Nacional de Promocion Cientifica y Tecnologica. PICT-2007-01050. Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnica. PIP 00363. Argentina. Universidad de Buenos Aires. M 043. Argentina.[br][br]Nothing to Disclose: NSB, MMB, VAL-L, CL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 144 69 650 SAT-569 PO24-01 Saturday 589 2012


590 ENDO12L_SAT-570 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Estrogen-Like Effects of Bisphenol A (BPA) on Estrogen Receptor (ER[alpha]) and Tumor Suppressor Protein p53 in Breast Cancer Cells Maria Yonan, Alessandra Boufford, Dana Ruskin, Lisa Shammas, Amy E Siebert, Amelita L Sanchez, G R Chaudhry, Sumi Dinda Oakland University, Rochester, MI; Oakland University, Rochester, MI; Oakland University, Rochester, MI Bisphenol A (BPA) is a polymerizing agent commonly found in plastics that has been linked to xenoestrogenic activity. We have studied tumor suppressor protein p53 and estrogen receptor (ER[alpha]) as possible molecular targets of BPA. Previous studies have indicated development of mammary and prostate tumors in rodents when prematurely exposed to BPA. In this study, we analyzed the estrogen-like effects of BPA on the expression of ER[alpha] and p53 with hormonal and anti-hormonal treatments in T47D breast cancer cells. Cells were cultured in medium containing 5% charcoal-stripped fetal bovine serum for 6 days in order to deplete any endogenous steroids or effectors. The cells were then treated for 24 h with 500 nM BPA, which was determined to be the optimal value by a concentration study of BPA from 1 nm to 2 [micro]M. Protein was extracted from the cells, quantified, and subjected to SDS-PAGE and Western blot analysis. For functional analysis, alterations in T47D cell proliferation were quantified upon exposure to BPA. Laser scanning confocal microscopy was performed to determine the cytolocalization of p53 and ER[alpha] upon treatment with BPA. Western blot analysis revealed that BPA caused an increase in the cellular protein p53 in a concentration dependent manner. However, cytolocalization of p53 upon treatment with estradiol and BPA remained unaltered. Also, increasing concentrations of BPA caused a 10-fold increase in cell number while BPA with ICI treatment caused a significant decrease in cell number. The levels of estrogen receptor (ER[alpha]) increased gradually in cells grown in charcoal-stripped media for 6-10 days. The presence of BPA had a modest effect of down regulation of ER[alpha] as compared to the control. These effects were sensitive to the presence of ICI. Our studies provide interesting leads to clearly delineate the possible mechanistic relationship among BPA, estrogen receptor, and tumor suppressor proteins in breast cancer cells.[br][br]Nothing to Disclose: MY, AB, DR, LS, AES, ALS, GRC, SD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1426 69 651 SAT-570 PO24-01 Saturday 590 2012


591 ENDO12L_SAT-571 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) BPA Exposure Affects Female Mouse Pubertal Development and G Protein Expression in the Hypothalamic-Pituitary-Gonadal Axis Scott Alan Sands, Tiffany Musick, Jared Leonard, Jill Diane Jacobson Children[apos]s Mercy Hospitals and Clinics, Kansas City, MO; University of Missouri-Kansas City School of Medicine, Kansas City, MO The earlier development of secondary sexual characteristics in girls in recent years suggests that factors other than genetics are involved, such as exposure to endocrine disrupting chemicals (EDC). One such EDC is bisphenol A (BPA), which is found in many common plastic products and whose distribution is ubiquitous. Puberty is controlled by GnRH, LH, and FSH. Receptors for these hormones couple to nucleotide binding proteins Gs[alpha] and Gq[alpha]. We have previously demonstrated that estrogen exposure upregulates expression of stimulatory G proteins. Thus, we hypothesized that exposure to BPA may result in altered timing of pubertal events, altered Gs[alpha] and Gq[alpha] Mrna expression in the mouse hypothalamic pituitary gonadal (HPG) axis, and/or altered sensitivity to GnRH. Prenatal treatment of pregnant mice with BPA (40 [micro]g/day) or vehicle (corn oil) began on gestational day 5, and continued every other day until parturition. Pubertal timing was assessed by monitoring vaginal opening from postnatal day 14 through 35. At day 28, mice underwent GnRH injections (100 [micro]g/ml) or vehicle (saline). RNA was extracted from hypothalamus, pituitary, and ovary and reversed transcribed into Cdna. In vitro studies involved exposure of the hypothalamic cell line GT1-7 and the gonadotrope cell line L[sup][szlig][/sup]T2, to BPA (10[sup]-6[/sup] to 10[sup]-8[/sup]M) for 96 hr. Quantitative PCR was performed using an iCycler. Results from RT-PCR were normalized using the housekeeping gene GAPDH and standardized using pooled control Cdna. Vaginal opening occurred earlier in BPA treated mice compared to vehicle (mean[plusmn] S.D.: 26.31 [plusmn] 3.46 days vs. 31.96 [plusmn] 3.38 days; p [lt] 0.001). Prenatal exposure to BPA led to increased expression of Gs[alpha] Mrna compared to vehicle in the hypothalamus (100 [plusmn] 24.84 vs. 208.35 [plusmn] 18.6; p [lt] 0.05) and ovary (100 [plusmn] 11.86 vs. 145.93 [plusmn] 38.8; p [lt] 0.05). Upon exposure to GnRH, pituitaries from mice prenatally exposed to BPA displayed an increase in Gs[alpha] Mrna compared to mice prenatally exposed to vehicle (508.59 [plusmn] 206.12 vs. 877.63 [plusmn] 136.92; p [lt] 0.05). In vitro BPA exposure (10[sup]-7[/sup]M) led to increased expression of Gs[alpha] Mrna (42.25 [plusmn] 10.7 vs. 62.25 [plusmn] 19.4; p [lt] 0.01) compared to vehicle in GT1-7 cells as well as in L[sup][szlig][/sup]T2 cells (149.7 [plusmn] 59.2 vs. 214.36 [plusmn] 21.0; p [lt] 0.01). These findings demonstrate that exposure to BPA is associated with earlier pubertal events, altered G protein Mrna expression in the hypothalamus and ovary, and altered responsiveness to GnRH.[br][br]Nothing to Disclose: SAS, TM, JL, JDJ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 539 69 652 SAT-571 PO24-01 Saturday 591 2012


592 ENDO12L_SAT-572 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Bisphenol-A Imprinting of GH-Dependent Sexually Dimorphic Liver Proteins and Genes in Female Rats Maria Cecilia Ramirez, Nadia Bourguignon, Maria Marta Bonaventura, Victoria Lux-Lantos, Carlos Libertun, Damasia Becu-Villalobos IBYME-CONICET, Capital Federal, Argentina Bisphenol A (BPA) is a known endocrine disruptor that is prevalent in our environment. As an estrogenic chemical, it has potential adverse effects on animals and humans exposed during embryonic developmental stage. In a previous work we demonstrated organizational effects of neonatal steroids on the GHRH-GH axis, and sexually dimorphic GH-dependent liver enzymes in females. While most studies on the effect of neonatal exposure to BPA have concentrated on the disruption of the reproductive axis, we test the hypothesis that GH imprinting of sexually dimorphic liver enzymes is also perturbed by neonatal BPA in female rats. On postnatal days 1-10, female pups received a daily sc. injection of BPA: 50 [mu]g/50 [mu]L (BPA50); 500 [mu]g/50 [mu]L (BPA500) or vehicle. Males were injected with castor oil. Rats were studied at 4 months of age. Serum IGF-I levels were not modified by neonatal BPA treatment, but pituitary GH concentration was higher in males compared to females (P=0.039) and not different from BPA treated females (P=0.11 and 0.46 males vs. BPA500 and 50, respectively). Liver IGF-I content, a downstream target of GH, was also higher in males compared to females (P=0.039), but not to BPA500 and 50 females (P=0.987 and P=0.259, respectively). We confirmed male specific liver expression of CYP2C11 mRNA levels, with a male/female ratio of 353. BPA treatment did not masculinize its expression in females. CYP2C12 mRNA was expressed predominantly in female rat livers (female/male ratio was 53). Neonatal BPA treatment induced a partial defeminization of this gene in female livers, as BPA50 and 500 females had significant different CYP2C12 mRNA levels from both males and females (P[lt]0.05, BPA50 and 500 vs. females and males). Female predominant genes (HNF-6 and ADH1) were also defeminized; while a male predominant liver protein (MUP) was not masculinized in females by neonatal BPA. We demonstrate that even though body weight, and serum IGF-I levels were not modified by neonatal BPA, pituitary GH content, and liver IGF-I concentration were increased, indicating partial masculinization of the GH axis. These changes produced a long-lasting defeminization of GH-dependent female predominant liver enzymes and transcription factors. Therefore, our findings should alert to the potential consequences of the in utero exposure to the endocrine disruptor as it may compromise liver metabolic enzymes, and modify therapeutic efficacy of drugs or susceptibility to systemic toxicity.[br][br]Nothing to Disclose: MCR, NB, MMB, VL-L, CL, DB-V 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 289 69 653 SAT-572 PO24-01 Saturday 592 2012


593 ENDO12L_SAT-573 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Urinary Phthalate Metabolite Concentrations Increase Odds of Impaired Fasting Glucose and Elevated HOMA-IR among Women in the National Health and Nutrition Examination Survey (NHANES) 2001[ndash]2008 Tamarra James-Todd, Elvira M Isganaitis, Aditi R Saxena Brigham and Women[apos]s Hospital, Boston, MA; Joslin Diabetes Center, Boston, MA Background: Phthalates are widely used chemical plasticizers that are ubiquitous in modern environments, and readily absorbed and metabolized by humans. Several studies suggest an association between phthalates and diabetes. Women have higher urinary concentrations of certain phthalate metabolites than men. We explored the association between urinary phthalate metabolites and fasting blood glucose and insulin resistance in women in a cross-sectional study.[br]Methods: We evaluated data for N=1927 females participating in the National Health and Nutrition Examination Survey (2001-2008). Urinary phthalate metabolites including mono-ethyl phthalate (MEP), mono-n-butyl phthalate (MnBP), mono-isobutyl phthalate (MiBP), mono-benzyl phthalate (MbzP), mono-(3-carboxypropyl) phthalate (MCP), and the sum of di-ethyl hexyl phthalate metabolites (DEHP), were analyzed by the CDC. Fasting blood glucose was measured (dichotomized as [ldquo]normal[rdquo], or less than 100 mg/dl vs. [ldquo]impaired[rdquo], or 100 to 126 mg/dl) and HOMA-IR was calculated based on fasting insulin and glucose (and dichotomized as greater or less than median HOMA-IR). We used multiple logistic regression, calculating odds ratios (OR) and 95% confidence intervals (CI), adjusting for urinary creatinine, sociodemographic and dietary factors. We excluded women with diabetes (glucose [gt]126 mg/dl) for analysis of fasting blood glucose, and self-reported diagnosis of diabetes for analysis of HOMA-IR.[br]Results: After adjusting for potential confounders, females within the highest quartile of MiBP had a 2.56-fold increased odds of impaired fasting blood glucose compared to females in the lowest quartile (95% CI: 1.20-5.43). Females in the third quartile of MnBP had 1.89-fold odds of higher than median HOMA-IR levels compared to females in the lowest quartile of MnBP (95% CI: 1.02-3.50). The highest quartile of MiBP conferred a 2.26-fold increased odds of higher than median HOMA-IR levels (95% CI: 1.08-4.73).[br]Discussion: Urinary concentrations of the phthalate metabolites MiBP and MnBP are associated with increased insulin resistance, as assessed by HOMA-IR, among U.S. females. MiBP was also associated with risk of impaired fasting glucose. These data provide support for the role of phthalates as endocrine disruptors of glucose homeostasis.[br][br]Nothing to Disclose: TJ-T, EMI, ARS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2134 69 654 SAT-573 PO24-01 Saturday 593 2012


594 ENDO12L_SAT-574 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Serum Di-Ethylhexyl Phthalate (DEHP) Levels in Obese Children Shin Hye Kim, Mi Jung Park Sanggye Paik Hospital, Inje University College of Medicine, Seoul, Korea [bold]Background:[/bold] Phthalates are synthetic chemicals produced in extremely large volumes for a wide variety of uses in personal care and consumer products, including building materials, food packaging, medical devices, toys and cosmetics. Though a few studies have shown that concentrations of phthalate metabolites are associated with obesity in adults, studies on association of phthalate concentrations with obesity in children are limited.[br][bold]Objectives:[/bold] We studied whether serum di(2-ethylhexyl) phthalate (DEHP) levels are associated with obesity in Korean children.[br][bold]Methods:[/bold] A total of 204 children (105 obese cased and 99 controls, aged 6 to 13yr) were enrolled. Nutrient intake, physical activity, household income, anthropometric characteristics, body composition analysis were measured. Fasting glucose, insulin, AST, ALT, uric acid and lipid profiles were determined, and serum DEHP levels were analyzed by gas chromatography/mass spectrometry method.[br][bold]Results:[/bold] Geometric mean serum DEHP levels were higher in obese children than in controls (53.8[plusmn]191.5 vs. 107.0[plusmn]341.5 ng/mL, P[lt]0.0001). In partial correlation analysis controlling for the potential covariates, serum DEHP level showed significant positive correlation with body mass index (r=0.194, P=0.015), serum ALT (r=0.159, P=0.047), uric acid (r=0.166, P=0.038) and body fat mass (r=0.170, P=0.029). Serum DEHP level didn[apos]t have significant correlation with HDL cholesterol, triglyceride, fasting blood sugar and fasting insulin. An increased risk of obesity [Odds ratio(95% Confidence Interval)] was observed according to the elevation of serum DEHP quartile in a dose-dependent manner [1.25(0.51-3.01) for quartile 2, 3.63(1.48-8.91) for quartile 3, 5.04(2.00-12.71) for quartile 4] after adjusting for age, gender, physical activity, household income and daily calorie intake.[br][bold]Conclusions:[/bold] Serum DEHP level was higher in obese children compared with controls, and it was associated with increased risk of obesity in a dose-dependent manner. Prospective studies are needed to determine potential causal links between DEHP exposure and childhood obesity.[br][br]Nothing to Disclose: SHK, MJP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 525 69 655 SAT-574 PO24-01 Saturday 594 2012


595 ENDO12L_SAT-575 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) A Cross Fostering Analysis of the Effects of PCB on Thyroid Status and Behavioral Development in Rat Pups David E Mankin, Zhi M Wang, Ellen N Dover, Howard C Cromwell, Lee A Meserve Bowling Green State University, Bowling Green, OH; Bowling Green State Univrsity, Bowling Green, OH Incorporation of PCB in the diet of pregnant rats has been shown to alter the thyroid status of offspring, as well as maternal behavior and pup behavior. In an effort to tease apart prenatal and postnatal PCB influence, in the present study pups either remained with their control (normal diet) or PCB fed (25 mg PCB 47/77 mixture/kg diet) dam, or were cross-fostered to a dam of the opposite dietary treatment. Rat pup behavior was determined on postnatal day (PND) 10 (ultrasonic vocalization, USV), PND 12-14 (conditioned odor preference for the dam, COP), PND 15-17 (grooming syntax), and PND 15-16 (open field behavior). Serum thyroxine concentrations were determined on PND 17. PCB significantly depressed thyroxine concentrations in a predictable fashion, with that of pups remaining with PCB dams depressed the most (35%), and those cross-fostered in either direction an intermediate amount (15%) as compared with pups remaining with control dams. Exposure to PCB increased the number per time of USV (69%) and line crosses in the open field (122%) regardless of cross fostering. Additionally, grooming behavior was disrupted most significantly on PND 16. Pups remaining with the PCB dams displayed the least success completing grooming chains (19%) followed by pups that were born to a PCB dam and then cross-fostered to a control dam (27%). Pups remaining with control dams had the greatest rate of grooming chain completion (42%). During COP testing, controls failed to show a preference for the presence or absence of a lemon scent paired with the dam. In contrast, pups exposed to PCB showed a strong aversion to lemon scent, spending only 29% of their time near the scent, even if it was paired with the dam. Although the results did not robustly separate prenatal and postnatal PCB exposure effects, taken together, the data support the contention that exposure to small amounts of PCB causes altered development of behavioral measures, regardless of either its influence on thyroid status or whether exposure occurs in the perinatal period during gestation only, lactation only, or both.[br][br]Sources of Research Support: Graduate College, Departments of Biological Science and Psychology, BGSU;.[br][br]Nothing to Disclose: DEM, ZMW, END, HCC, LAM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 768 69 656 SAT-575 PO24-01 Saturday 595 2012


596 ENDO12L_SAT-576 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Changes in Concentrations of PCBs and PCDD/Fs in Breast Milk from Hungary during the First Three Months of Lactation Janos Garai, Andrea Colombo, Emilio Benfenati, Helen Hakansson, Marika Berglund, Jozsef Bodis, Eva Vigh Medical School, University of P[eacute]cs, P[eacute]cs, Hungary; Instituto di Richerche Farmacologiche [quot]Mario Negri[quot], Milan, Italy; Karolinska Institutet, Stockholm, Sweden; Medical School, University of P[eacute]cs, P[eacute]cs, Hungary Human exposure to toxic persistent organic pollutants (POPs) such as polychlorodibenzodioxins (PCDDs), polychlorodibenzofurans (PCDFs) and polychlorobiphenyls (PCBs) occurs essentially via food consumption. These contaminants are transferred to breast milk, therefore breastfed infants are at risk of being exposed to considerable amounts of POPs during this sensitive age. In the present study individual breast milk samples were collected at three time points (postpartum day 5, 12 and 84) from 34 mothers who delivered their infants during 2007 in Baranya County, Hungary. Breast milk samples were analyzed for 17 PCDD/Fs, 12 dioxin-like (DL) PCBs and 7 non-dioxin-like (NDL) PCBs using high-resolution gas chromatography/high-resolution mass spectrometry. A declining trend for most congeners was observed during lactation with a main decrease between days 5 and 12. The total toxic equivalent (TEQ) concentrations, derived from PCDD/Fs and DL-PCBs, were 2.95 [plusmn] 1.59 pg-TEQ/g fat, 2.63 [plusmn] 1.52 pg-TEQ/g fat and 2.40 [plusmn] 1.45 pg-TEQ/g fat at the three time points, respectively. The corresponding NDL-PCB concentrations were 32 [plusmn] 30, 27 [plusmn] 20 and 27 [plusmn] 23 ng/g fat, respectively. The results highlight the importance of timing of breast milk sampling for exposure assessment. Levels of pollutants in Hungarian breast milk samples were in the low limit of concentration range for Europe. This is the first study which presents data on POPs in individual Hungarian breast milk samples.[br][br]Sources of Research Support: The work was performed within the EU FP6 funded CASCADE Network of Excellence (Contract No. FOOD-CT-2004-506319); The authors are grateful to all mothers who kindly accepted to participate in this study.[br][br]Nothing to Disclose: JG, AC, EB, HH, MB, JB, EV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1504 69 657 SAT-576 PO24-01 Saturday 596 2012


597 ENDO12L_SAT-577 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Persistent Organic Pollutant (POPs) Levels in Human Visceral and Subcutaneous Adipose Tissue in an Obese Portuguese Population [mdash] Biological Implications Diogo Pestana, Carla Sa, Virginia Fernandes, Gil Faria, Diana Teixeira, Ana Faria, Manuela Meireles, Sonia Norberto, Ana Cunha, Edgar Moreira, Bruno Santos, Sonia Martins, Ana Cristina Santos, Rosario Monteiro, Valentina Domingues, Cristina Delerue-Matos, Conceicao Calhau Faculty of Medicine, University of Porto, Porto, Portugal; Faculty of Sciences, University of Porto, Porto, Portugal; Instituto Polit[eacute]cnico do Porto, Porto, Portugal; S Jo[atilde]o Hospital, Faculty of Medicine, University of Porto, Porto, Portugal; Faculty of Medicine and Institute of Public Health, University of Porto, Porto, Portugal There are extended evidences associating persistent organic pollutants (POPs) and obesity etiology. Due to their lipophilicity, persistence and bioaccumulative nature, adipose tissue (AT) is a common site of POPs accumulation. In this regard, we aimed to evaluate POPs presence in human AT in an obese population subjected to bariatric surgery, and assess its putative association with metabolic parameters.[br]Adipose tissue samples (visceral, vAT and subcutaneous, scAT; n=88) from an obese Portuguese population (body mass index, BMI [gt] 40) were collected during surgery at the Hospital S. Jo[atilde]o (protocol approved by the Hospital Ethics Committee). The levels of 13 POPs residues were determined by GC-ECD.[br]All human samples, visceral and subcutaneous AT, evaluated had detectable levels of POPs. The median concentration of total POPs was higher in older women (46-62 years), and visceral AT had a higher concentration of POPs than subcutaneous AT. Regarding the size of adipocytes, the mean area was significantly higher in samples with higher concentration of POPs. This positive correlation was observed in visceral and subcutaneous AT. Comparing with comorbilities, the subjects with higher concentrations of POPs had more prevalence of type 2 diabetes (not significant), dyslipidemia and hypertension.[br]These preliminary results confirm that POPs are pervasive in this obese population, their abundance increasing with age. Their putative association with metabolism and outcome after surgery will be investigated.[br][br]Sources of Research Support: This work was supported by FCT (POCI, FEDER, Programa Comunit[aacute]rio de Apoio, PTDC/QUI/65501/2006; SFRH/BD/46640/2008, SFRH/BD/47200/2008, SFRH/BD/64691/2009, and SFRH/BPD/40110/2007) and [ldquo]Projectos de Investiga[ccedil][atilde]o na Pr[eacute]-gradua[ccedil][atilde]o 2010, Universidade do Porto-150.[br][br]Nothing to Disclose: DP, CS, VF, GF, DT, AF, MM, SN, AC, EM, BS, SM, ACS, RM, VD , CD-M, CC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2008 69 658 SAT-577 PO24-01 Saturday 597 2012


598 ENDO12L_SAT-578 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Evaluation of the Effects of Estrogenic Ligands on Transcriptional Activation Mediated by Zebrafish Estrogen Receptors Using Reporter Cell Lines Caroline Pinto, Marina Grimaldi, Abdel Boulahtouf, Maria Bondesson, Jan-Ake Gustafsson, Patrick Balaguer University of Houston, Houston, TX; INSERM U896, Montpellier, France Zebrafish ([italic]Danio rerio[/italic]) has been used as an alternative [italic]in vivo [/italic]model for the study of the effects of endocrine disrupting chemicals, including xenoestrogens, that interfere with normal developmental and physiological processes. Even though only two estrogen receptors (ER) subtypes have been characterized in mammals (ER[alpha] and ER[beta]), the presence of three ER forms in zebrafish and most teleosts have been reported: ER[alpha] (esr1), ER[beta]1 (esr2b) and ER[beta]2 (esr2a). However, little is known about the activity and transcriptional profiles of known mammalian estrogenic ligands in the zebrafish ERs (zfERs). The purpose of this study was to evaluate the effects of known endogenous, environmental and pharmaceutical (anti)estrogenic compounds on the transcriptional activation of the zfERs using HELN-zfER[alpha], HELN-zfER[beta]1 and HELN-zfER[beta]2 cell lines, which stably express an ERE-driven luciferase reporter and the full-length ER[alpha], [beta]1 and [beta]2, respectively. We report that the bisphenols BPA, BPAF and BPC have a higher zfER[alpha] agonist potency relative to the zfER[beta] subtypes. The endogenous estrogens estrone, estriol and 17[alpha]-E2 show stronger potency in transactivating zfER[alpha] and zfER[beta]1 compared to the zfER[beta]2 isoform, whereas 17[beta]-E2 displays a slight zfER[beta]1 selectivity relative to the other zfER subtypes. Among all the compounds tested, the human ER[alpha] selective compound 16[alpha]-LE2 demonstrated the best selectivity towards its zfER counterpart, with approximately 365 and 100-fold higher potencies to transactivate zfER[alpha] compared to zfER[beta]1 and zfER[beta]2, respectively. PPT, another human ER[alpha] selective compound, was surprisingly devoid of zfER[alpha] agonistic activity and instead displayed antagonistic activity. Moreover, the human ER[beta] selective pharmaceuticals 8[beta]VE2 and DPN also showed preferential selectivity for zfER[alpha] rather than for the zfER[beta] isoforms. Our results suggest that the transcriptional activity of a given ligand can differ between the human ERs and their zebrafish counterparts, and thus, the transcriptional selectivity of the compounds should be tested prior to their [italic]in vivo [/italic]use. Since zebrafish is used as a model for the study of the effects of xenoestrogens, determining the transcriptional profiles of estrogenic compounds in the zfERs is crucial to apply the zebrafish model to ER-related studies and their extrapolation to the mammalian system.[br][br]Nothing to Disclose: CP, MG, AB, MB, J-AG, PB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1605 69 659 SAT-578 PO24-01 Saturday 598 2012


599 ENDO12L_SAT-579 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Zebrafish Embryos as a Model for Studying Estrogen Disruption Ruixin Hao, Maria Bondesson, Patrick Balaguer, Jan-Ake Gustafsson University of Houston, Houston, TX; Institut de Recherche en Canc[eacute]rologie de Montpellier, Montpellier, France Estrogenic endocrine disruptor exposure during critical developmental windows in early life may cause health concerns such as abnormal sexual differentiation, infertility, metabolic disorders, obesity and certain types of cancer later in life. As the estrogen signaling pathways are conserved from fish to mammals, zebrafish have become an emerging model for studying estrogen disruption. Features like a high number of offspring, ex-utero and fast embryonic development, and small size facilitating high throughput screening, have promoted their use in toxicology. We investigated estrogen signaling during early zebrafish embryonic development by performing whole genome expression analysis. Zebrafish embryos were exposed to estrogen from 3 hours post fertilization (3 hpf) to 4 days post fertilization (4 dpf). Embryos were harvested daily and RNA was extracted for microarray analysis. Known estrogen target genes, such as vitellogenins and cyp19a1b, were strongly up-regulated by estrogen treatment at 4 dpf. Several new up-regulated and down-regulated genes were identified and validated using real-time PCR. Furthermore, the new target genes were tested as potential biomarkers of estrogen exposure to analyze whether endocrine disruptors such as bisphenol A, C and AF affect estrogen signaling during early development. The results show that the estrogen disruptors activated or inhibited estrogen signaling in a target gene specific manner. The new biomarkers will be used to screen a battery of endocrine disruptors for interference with estrogen signaling during early development.[br][br]Sources of Research Support: This work was supported by grants from the Emerging Technology Grant from the Office of the Governor of Texas to the Center of Nuclear Receptors and Cell Signaling, and from EPA (R834289).[br][br]Nothing to Disclose: RH, MB, PB, J-AG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1124 69 660 SAT-579 PO24-01 Saturday 599 2012


600 ENDO12L_SAT-580 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Use an Avian Cell Culture Model To Analyze Effects of Endocrine Disruptors on the Testicular Tissue Pascal Froment, Edith Guibert, Berenice Prieur, Jean-Pierre Brillard INRA - PRC - Station de Recherche Avicole, Nouzilly, France; Fertil[apos]avi, Rouziers De Touraine, France In recent decades, many tests in vivo or in vitro models mainly based on mammalian (rat-mouse) and fish were used to assess the risks raised by contact or ingestion of molecules with an pharmaceutical, agricultural or natural origin. But, none or few in vitro tests using bird models have been explored despite their advantages: the embryonic gonads of birds have a high plasticity of development, and sperm production is nearly two times faster than in rodents. Thus, birds present one gonad in female and two gonads in male. In the presence of estrogenic substances or antagonist of estrogen production, during the first days of the embryonic life, the sex gonadic phenotype could change.[br]Hence, as part of a national project (post-Grenelle Environment Forum), we have established the in vitro culture of germ cells and somatic cells from chicken testis and we have evaluated the sensitivity against two molecules known to be endocrine disruptors: phtalates (mono-(2-ethylhexyl) phthalate, MEHP) and ethinyl estradiol (EE2) in comparison to previous studies using rodent and human system.[br]After 96 h of culture in presence of 10[mu]M MEHP, cultures present a structural alteration, a reduction in cell proliferation and in germ cells population. These results are in accordance with previous studies using rat, mice or humans culture of testicular cells and with similar sensitivity levels or even better sensitivity for some [apos]end-points[apos] (biological parameters). Moreover, we observed an increase in proliferative germ cells and a decrease in apoptosis or oxydative stress starting at the lower EE2 dosis (0.03 ng/ml, output value of wastewater) to 300 ng/ml.[br]These cells preparations were also able to be frozen and thawed for a latter use, because VASA-positive germ cells were detected after 96h of culture. In conclusion, the establishment of this testicular cells culture could be considered as an in vitro alternative method, complementary to mammals model, limiting the number of animals used and with an elevated sensitivity at least for these two endocrine disruptors.[br][br]Sources of Research Support: This work was supported by Institut National de la Recherche Agronomique, INRA, and by the French Ministry of Environment in a national program [quot]Grenelle de l[apos]Environnement programme LOLF 189 funding (Specific Tools and Methods for Reprotoxicity program).[br][br]Nothing to Disclose: PF, EG, BP, J-PB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1706 69 661 SAT-580 PO24-01 Saturday 600 2012


601 ENDO12L_SAT-581 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Prevalence of Phenotypes and Etiologies of Hypospadias Evaluated at the Center of Gender Medicine at Texas Children[apos]s Hospital Min-Jye Chen, Lefkothea Karaviti, Lynda Jacks, O[apos]Brian Smith, Jennifer Dietrich, Marni Axelrad, David Roth, Reid Sutton, Sheila Gunn Texas Children[apos]s Hospital, Houston, TX; Baylor College of Medicine, Houston, TX; Texas Children[apos]s Hospital, Houston, TX; Texas Children[apos]s Hospital, Houston, TX; Texas Children[apos]s Hospital, Houston, TX; Baylor College of Medicine, Houston, TX; Texas Children[apos]s Hospital, Houston, TX Background: Hypospadias is one of the most frequent congenital malformations in males, affecting approximately 20-40 in 10,000 live births. A review of previous studies indicates a multifactorial basis for the condition, including hormonal, genetic, and environmental causes.[br]Objective: The aim of this study was to characterize hypospadias in patients seen in the multidisciplinary Gender Medicine clinic at Texas Children[apos]s Hospital. We hypothesize that the exact etiology of hypospadias will remain unknown in most patients.[br]Methods: A retrospective chart review was performed on 38 patients with hypospadias seen in the Gender Medicine clinic between January 2006 and October 2011. Degree of hypospadias was classified as glandular (Grade 1), coronal (Grade 2), penile (Grade 3), penoscrotal (Grade 4), scrotal (Grade 5), or perineal (Grade 6). Etiology was classified as either 1) a genetic association 2) a hormonal defect 3) a prenatal exposure 4) Developmental field defect or 5) a yet unknown etiology.[br]Results: Of these 38 patients, hypospadias severity was noted as follows: No patients had Grade 1; 1 (3%) had Grade 2 (95% CI 0-12%); 7 (18%) had Grade 3 (8-33%); 21 (55%) had Grade 4 (39-70%); 6 (16%) had Grade 5 (6-30%); and 3 (8%) had Grade 6(2-20%). Of all the patients, 39% (25-55%) had a known genetic association, 8% (2-20%) were determined to have a developmental field defect, and 53% (37-68%) had hypospadias of yet unknown etiology. No patients had identified hormonal or environmental causes; however, one patient in the field defect group (VACTERL) and with penile hypospadias (Grade 3) had a maternal occupational exposure to laboratory chemicals. When patients were plotted on a map based on zip codes, a cluster of 11 patients (29%) was noted within an approximately 4 mile radius in Southwest Houston (95% CI [16-45%]).[br]Conclusion: The majority of patients were found to have more severe hypospadias than in the general population as per the studies by Paulozzi and Nassar, likely due to more severe case referrals. Known genetic etiology in our patients was associated with greater incidence of penile and penoscrotal hypospadias, when compared to the other groups.[br]Fifty-three percent of patients had an unknown cause for their hypospadias. In these cases, environmental factors may be implicated. Future studies will be undertaken to evaluate this and also the significance of the clustering of hypospadias patients within one specific area of Houston.[br][br]1. Toppari J, Virtanen HE, Main KM, Skakkebaek NE. Cryptorchidism and hypospadias as a sign of testicular dysgenesis syndrome (TDS): environmental connection. Birth Defects Res A Clin Mol Teratol. Oct 2010;88(10):910-9. 2. Boisen KA, Chellakooty M, Schmidt IM, Kai CM, Damgaard IN, Suomi AM, Toppari J, Skakkebaek NE, and Main KM. Hypospadias in a cohort of 1072 danish newborn boys: prevalence and relationship to placental weight, anthropometrical measurements at birth, and reproductive hormone levels at three months of age. J Clin Endocrinology Metab. July 2005;90(7):4041-4046. 3. Paulozzi LJ, Erickson JD, Jackson RJ. Hypospadias trends in two US surveillance systems. Pediatrics.1997;Nov100(5):831-4. 4. Nassar N, Bower C, Barker A. Increasing prevalence of hypospadias in Western Australia, 1980-2000. Arch Dis Child. 2007;Jul 92(7):580-4. Epub 2007 Apr 3.[br][br]Nothing to Disclose: M-JC, LK, LJ, OS, JD, MA, DR, RS, SG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2242 69 662 SAT-581 PO24-01 Saturday 601 2012


602 ENDO12L_SAT-582 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Oxidative Stress in Rat Testis Tissues Exposed to Plant Growth Regulator (4-Chlorophenoxy Acetic Acid) Ediz Yesilkaya, Cigdem Ozer, Aysun Bideci, Peyami Cinaz Gulhane Military Medical Academy, Ankara, Turkey; Gazi University, Ankara, Turkey; Gazi University, Ankara, Turkey [italic]Introduction:[/italic] Plant Growth Regulators are generally used in greenhouses to obtain maximum yield; 4-Chlorophenoxy acetic acid being the most widely used one. Plant Growth Regulators remnants may lead to adverse effects at inappropriate or high doses. Hepatocellular necrosis and/or increased apoptosis in genital organs have been reported in rats exposed to this substance. [italic]Objectives:[/italic] We aimed to investigate the possible role of oxidative mechanisms in rat testis exposed to 4-chlorophenoxy acetic acid. [italic]Methods: [/italic]The study was implemented on 20 day-old Wistar albino rats. Forty rats were randomized into five groups (a control group, a saline group and three 4-Chlorophenoxy acetic acid groups that received 25-50-100 mg/kg/day until 50 days of age respectively). [italic]Results:[/italic] There was no statistical significant difference between saline and control group in terms of in terms of oxidative stress markers (MDA, GSH and NOx levels) whereas there was a significant difference between 4-Chlorophenoxy acetic acid received group and control group in terms of MDA and NOx levels. [italic]Conclusions:[/italic] 4-Chlorophenoxy acetic acid may have an impact on rat testis at tissue level by oxidative stress and may eventually lead to infertility.[br][br]Nothing to Disclose: EY, CO, AB, PC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 111 69 663 SAT-582 PO24-01 Saturday 602 2012


603 ENDO12L_SAT-583 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Disruption of Androgen Receptor Function by Brominated Compounds Joubert Banjop Kharlyngdoh, Solomon Asnake, Ajay Pradhan, Huthayfa Mujahed, Per-Erik Olsson Orebro University, Orebro, Sweden With the development of new biological and chemical analyses there is an increased possibility to identify adverse effects of complex and low-level exposures on the health and well being of both humans and wildlife. We are using a combination of molecular modeling and experimental analysis to screen for brominated compounds that interact with androgen receptor (AR) signaling. We have earlier identified 1,2-dibromo-4-(ethyl)-cyclohexane (TBECH) as a group of potent agonists to the human AR (hAR). TBECH exist as 4 isomers with corresponding enantiomers. The potency of TBECH isomers to activate hAR is [delta][gt][gamma][gt][alpha][gt][beta]. However, hAR with the T877A mutation, commonly found in prostate cancer, results in reduced discrimination between the isomers and higher overall activation of hAR. We show that TBECH and its isomers also induce AR activation in chicken and zebrafish, although the response is much weaker in these two species than in humans. Further to the identification of TBECH as an androgen agonist we have now identified several additional brominated compounds as androgen antagonists in mammals, birds and fish. These compounds include Ambroxol, a pharmaceutical used in treatment of Cystic Fibrosis patients and the brominated flame retardant, 4,5,6,7-tetrabromo-1,1,3-trimethyl-3-(2,3,4,5 tetrabromophenyl)-lindan (OBIND). Using qPCR arrays we have also obtained results that indicate complex and cell specific responses of some androgen antagonists, including regulation of AR transcripts. Using a combination of molecular modeling, receptor activation reporter systems and gene activation assays we are presently screening for other brominated flame-retardants that have possible interactions with AR from different species.[br][br]Nothing to Disclose: JBK, SA, AP, HM, P-EO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1614 69 664 SAT-583 PO24-01 Saturday 603 2012


604 ENDO12L_SAT-584 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Triclocarban (TCC) Exposure in Pregnancy and during the Female Neonate Period Compromises Lactation and Reproductive Development Rebekah C Kennedy, Jiyoung Bae, Kellie Fecteau, Ling Zhao, Nancy A Gee, Kurt Benirschke, Bill L Lasley, Jiangang Chen University of Tennessee, Knoxville, TN; University of Tennessee, Knoxville, TN; University of Tennessee, Knoxville, TN; University of California, Davis, CA; University of California, San Diego, CA Triclocarban (TCC), an antimicrobial compound in personal care products, has been shown to disrupt endocrine function in vitro and in vivo in adult male rats by influencing sex hormone dependent transcriptional activity and organ growth. However, whether or not TCC exposure in utero alters the trajectory of subsequent life stages is not known. This study investigates the relationship between TCC exposure during pregnancy and neonatal development using an in vivo rat model. Pregnant Sprague Dawley (SD) rats were provided ad lib rat chow supplemented with 0.5% w/w TCC from gestational day 4 until sacrifice at weaning/postnatal day (PND) 21. Sham controls received unsupplemented rat chow only. Upon weaning, treatment was continued in the surviving female neonates until at least one diestrus cycle was observed. Neonate number after delivery and pup mortality throughout the study were recorded. Systemic TCC levels of both dams and neonates were determined and sex organs were weighed and fixed. The anal-genital distance (AGD) of neonates was recorded until vaginal opening (VO). We found that none of the neonates born to the TCC exposed dams survived beyond PND 8 due to apparent inadequate milk production by their dams. Histological evaluation of breast tissue from TCC treated dams revealed massive and destructive inflammation, which stopped milk production and indicate a compromised immune system. These observations are consistent with in vitro data using RAW 264.7 macrophage cells, which demonstrate that TCC modulates the lipopolysaccharide (LPS) induced NF-[kappa]B activity, the immediate early activator of genes involved in the inflammatory and immune response. Since no neonates of TCC treated dams survived, half of the female neonates from the control dams were switched to 0.5% w/w TCC supplemented diet on PND 21with food intake and body weight recorded daily. TCC treatment did not affect body weight gain or food intake of neonates. The mean liver weight of the TCC treated group was 28% higher per gram body weight when compared to controls. Moreover, a significant delay in the onset of puberty, as measured by VO (39.5[plusmn]1.0 vs. 43 days), was observed in treated neonates. These results demonstrate that in utero and neonatal TCC exposure can adversely affect reproductive development and provide rationale for further investigations of the potential adverse effects from TCC exposure on the integrity of the immune system.[br][br]Sources of Research Support: This study was partially supported by NIH 5R21 ES0117475-02 and 1R21 ES016802.[br][br]Nothing to Disclose: RCK, JB, KF, LZ, NAG, KB, BLL, JC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 210 69 665 SAT-584 PO24-01 Saturday 604 2012


605 ENDO12L_SAT-585 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) The Endocrine-Disrupting Chemical Tolylfluanid Disrupts Adipocyte Metabolism Via Glucocorticoid Receptor Activation Brian A Neel, Robert M Sargis, Matthew J Brady University of Chicago, Chicago, IL; University of Chicago, Chicago, IL Emerging data implicate environmental endocrine disrupting chemicals (EDCs) as contributors to the burgeoning global obesity and diabetes epidemics. These environmental toxins disrupt normal hormonal signaling in a manner that promotes excess adiposity and disturbs normal metabolism. The central role of adipose tissue in systemic energy metabolism and the bioaccumulation of lipophilic EDCs in the adipocyte lipid droplet make adipose tissue an important model to study the metabolic effects of EDCs. In an initial screen of putative metabolic disruptors, the fungicide tolylfluanid (TF) was found to robustly inhibit insulin signaling and glucose uptake in the 3T3-L1 adipocytes. Further analyses showed that TF inhibits insulin-stimulated Akt phosphorylation in primary murine and human adipocytes. In an interrogation of the insulin signaling pathway, TF inhibited PI3Kinase association with insulin receptor substrate-1 (IRS-1) through a downregulation of IRS-1 transcript and protein levels at concentrations from 10 nM to 1 [mu]M after 24 hours in culture. This attenuation of insulin signaling led to an inhibition of leptin secretion as well as a broader shift in the adipokine secretory profile. The next aim was to determine the molecular mechanism of this alteration in insulin signaling. Previous work has shown that many EDCs bind to and activate various nuclear receptors. Because the glucocorticoid receptor (GR) is known to modulate adipocyte physiology, the effect of TF on GR activation was examined. Using the glucocorticoid-induced leucine zipper (GILZ) gene as a readout of GR activation, TF exhibited glucocorticoid-like activity, with IRS-1 expression correlating with the degree of GR activity. Interestingly, TF mimicked glucocorticoid action by acutely increasing and chronically decreasing insulin responsiveness through changes in IRS-1 levels. At equal levels of GR activity, TF increased insulin-stimulated lipogenesis to a similar extent as the active murine glucocorticoid corticosterone. Cellular fractionation assays also demonstrated that TF promotes the activating phosphorylation of GR, driving it from the cytoplasm into the nucleus. Collectively, these results demonstrate the ability of TF to inhibit insulin signaling through a specific downregulation of IRS-1, likely through GR activation. This novel molecular mechanism of EDC action supports a potential role for these chemicals in the pathogenesis of obesity and diabetes.[br][br]Sources of Research Support: The National Institutes of Health [T32-HL007237 supporting B.A.N.; K08-ES019176 and F32-ES017397 to R.M.S.] and by the Diabetes Research and Training Center [P60-DK020595].[br][br]Nothing to Disclose: BAN, RMS, MJB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1360 69 666 SAT-585 PO24-01 Saturday 605 2012


606 ENDO12L_SAT-586 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Exposure to Inorganic Arsenite Alters Glucose Metabolism of Pregnant Rats Maria M Bonaventura, Nadia S Bourguignon, Carlos Libertun, Victoria A Lux-Lantos Instituto de Biologia y Medicina Experimental-CONICET, Buenos Aires, Argentina; Universidad de Buenos Aires, Buenos Aires, Argentina; Universidad de Buenos Aires, Buenos Aires, Argentina Inorganic arsenic is a worldwide distributed natural contaminant. The most common source of contamination is from ground water. Inorganic arsenic is highly toxic and a proved carcinogenic in humans. Recently it has also been described as an endocrine disruptor impacting the metabolic and reproductive axes, although the mechanisms have not been elucidated.[br]Female Sprague-Dawley rats were treated with sodium arsenite in drinking water (5 or 50 ppm in distilled water or distilled water as control) from gestation day 1 (GD1) (confirmed by presence of vaginal sperm plug) to sacrifice (two months post-partum). Standard chow was given ad libitum. Body weight (BW) was recorded during pregnancy and on GD15-17, a glucose tolerance test (GTT) was performed. Additionally, dams were evaluated by a GTT one month after weaning. For GTTs, intraperitoneal glucose (2 g/kg BW) was injected to overnight-fasted rats and blood glucose levels were evaluated at different time points. On postnatal day 1 (PND1), pups were counted, sexed and weighted. Results are expressed as means [plusmn] S.E.M. Differences between means were analyzed by one-way ANOVA or two[ndash]way ANOVA with repeated measures design. P[lt]0.05 was considered statistically significant.[br]No differences were found on BW during pregnancy. The GTT was significantly altered in pregnant animals treated both with 5 or 50 ppm of sodium arsenite (A5 and A50, respectively), being the glycemia at 30 minutes post glucose overload significantly higher in treated animals compared to controls (mg/dl: C: 171 [plusmn] 10, A5: 281 [plusmn] 21, A50: 290 [plusmn] 22; A50, A5 vs. C, p[lt]0.001, n= 6-8). However, the GTT performed one month after weaning showed no differences between groups. Dams from all groups delivered their offspring at 23 days and no differences were observed on litter size and litter sex. BW of pups on PND 1 was significantly diminished in the A50 pups from both sexes, compared with controls; no differences were observed in A5 treated animals [BW (g): Female pups: C: 7.7 [plusmn] 0.1, A5: 7.9 [plusmn] 0.1, A50: 7.3 [plusmn] 0.1; A50 vs. C, A5, p[lt]0.05, n=39 for each group; Male pups: C: 8.1 [plusmn] 0.1, A5: 8.1 [plusmn] 0.1, A50: 7.6 [plusmn] 0.1; A50 vs. C, A5 p[lt]0.005. n= 31-40].[br]We conclude that exposure to sodium arsenite during pregnancy produces a clear alteration in glucose homeostasis on dams, probably inducing the alterations on body weight observed in their pups. These results suggest that inorganic arsenic could alter the metabolism of both dams and their offspring.[br][br]Sources of Research Support: Agencia Nacional de Promocion Cientifica y Tecnologica. PICT-2007-01050. Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. PIP 00363. Argentina. Universidad de Buenos Aires. M043. Argentina.[br][br]Nothing to Disclose: MMB, NSB, CL, VAL-L 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 214 69 667 SAT-586 PO24-01 Saturday 606 2012


607 ENDO12L_SAT-587 POSTER SESSION: Phytoestrogens [amp] Endocrine Disrupting Chemicals (1:30 PM-3:30 PM) Neonatal Exposure to Perfluorooctane Sulfonate (PFOS) and Perfluorooctanoic Acid (PFOA) Causes Early Puberty Onset and Elevated Postpubertal Hormone Levels in Female Rats Guizhen Du, Jialei Hu, Ling Song, Xinru Wang, Di Wu Institute of Toxicology, Nanjing Medical University, Nanjing, China; School of Public Health, Nanjing Medical University, Nanjing, China Perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) have been reported to have endocrine disruptive properties. To determine the effects of PFOS and PFOA on female reproductive maturation, we recorded the puberty onset and postpubertal estrous cycles in female Sprague-Dawley rats after neonatal exposure to PFOS and PFOA. Newborns were cross-fostered and given PFOS or PFOA by subcutaneous injection at three concentrations (0.1mg/kg, 1mg/kg and 10 mg/kg) for 5 days (postnatal day P1-5) (9 female rats per group). Anogenital distance (AGD) and body weight on birthday, P15, P25 and P35 were measured. Ages of eye opening and vagina opening were recorded. Postpubertal estrous cyclicity was examined based on vaginal cytology. Animals were perfused at proestrus following two complete cycles or on the 15th day after vagina opening. A blood sample was collected from the anesthetized rat by cardiac puncture just prior to the perfusion. Ovarian mass was measured and serum estrodial, luteinizing hormone (LH) were detected. The results showed that compared to the control, vaginal opening were significantly advanced in 1 mg/kg PFOS (p[lt]0.01), 10 mg/kg PFOS (p[lt]0.05) and 10 mg/kg PFOA (p[lt]0.01) groups. Serum LH level was significantly higher in the 10 mg/kg PFOA group compared to the control (P[lt]0.05). Treatments of 0.1mg/kg, 1mg/kg PFOS and 0.1 mg/kg, 1mg/kg PFOA significantly elevated estradiol level (all p[lt]0.01). Interestingly, high-dose treatment 10 mg/kg PFOS and 10 mg/kg PFOA groups didn[apos]t show any difference in serum estradiol concentration compared to the control. Vaginal cytology showed all stages of estrous cyclicity in each group and no difference was found. There was no significant difference in ovary weight and eye-opening age. AGD values were normalized to the cubed root of body weight for analysis.AGD and body weight at birth didn[apos]t show any difference among groups. However, the 0.1 mg/kg PFOA group were found to have bigger AGD on P25 and P35 (p[lt]0.001, p[lt]0.01). Groups of 0.1 mg/kg and 10mg/kg PFOS had bigger AGD on P35 compared to the control (p[lt]0.05, P[lt]0.001). Rats in 10mg/kg PFOA and 10mg/kg PFOS groups were significantly heavier on P15 (both p[lt]0.01), P25 (p[lt]0.05, p[lt]0.01) and P35 (both p[lt]0.05). 0.1mg/kg PFOS treatment significantly elevated body weight only on P15 (p[lt]0.05). These results indicate that both PFOS and PFOA have the capacity to disturb the normal female reproductive maturation and physiology in a non-dose-response manner.[br][br]Sources of Research Support: National Natural Science Foundation of China 81102147.[br][br]Nothing to Disclose: GD, JH, LS, XW, DW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 765 69 668 SAT-587 PO24-01 Saturday 607 2012


608 ENDO12L_SAT-588 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) The Comparison Study of Urinary Iodine Excretion between Prepubertal Children with and without Simple Goiter Sun Hee Lee, Min Jae Kang, Hye Rim Chung, Jae Hyun Kim, Choong Ho Shin, Sei Won Yang Busan Paik Hospital, Busan, Korea; Seoul National University, Seoul, Korea; Seoul National Univertisy Bundang Hospital, Seongnam-si, Korea; Ilsan Paik Hospital, Goyang-si, Korea [bold]Purpose[/bold]: Simple goiter is defined as a thyroid enlargement without inflammation or neoplasm in an individual, whose thyroid function is euthyroid. Iodine excess or iodine deficiency is regarded as the most important environmental factor of the simple goiter. However, there have been a few reports on the research of iodine excess, whereas there have been many reports on the researches about the iodine deficiency in children with goiter. Moreover, there has been no data about the iodine intake status of Korean children. This study aimed to assess the iodine intake status of prepubertal Korean children by measurement of urinary iodine (UI) excretion and evaluate whether iodine status affects on the development of simple goiter.[br][bold]Patients and Methods[/bold]: This study included prepubertal children who visited endocrinology clinic of Seoul National University Children[apos]s Hospital between June 2009 and September 2010. The participants were divided into simple goiter group and health control group. Thyroid function tests were done to all subjects and thyroid antibody tests were examined to simple goiter group. Only children, who showed euthyroid state and negative results to thyroid antibody tests, were included. UI excretion was checked using the morning spot urine. Familial thyroid disease history and second hand smoking history measured.[br][bold]Results[/bold]: Total 69 prepubertal children were included in the research (28 simple goiter group and 41 health control group). The ratio of male and female was 1:2.1 in simple goiter group. There were no significant differences in age, free thyroxine level, thyroid stimulating hormone, prevalence of familial thyroid disease history and second hand smoking history between two groups. The UI level did not reveal any significant difference between two groups (simple goiter group 1202.70[plusmn]1493.38 [mu]g/L vs. health control group 933.88[plusmn]1182.42 [mu]g/L, P=0.191). The UI excretion of all subjects was 1042.97[plusmn]1313.63 [mu]g/L. The prevalence of iodine excess (UI[ge]300 [mu]g/L) was 65.2%.[br][bold]Conclusion[/bold]: This study suggests that iodine excess did not influence on simple goiter directly in Korean prepubertal children. However, this study revealed that iodine intake levels in Korean prepubertal children were very high and the majority of children in the study were iodine excess state. Further longitudinal studies are needed to investigate the role of iodine excess on the change of thyroid function or the development of thyroid disease.[br][br]Nothing to Disclose: SHL, MJK, HRC, JHK, CHS, SWY 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 864 70 669 SAT-588 PO30-01 Saturday 608 2012


609 ENDO12L_SAT-589 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) A New PAX8 Mutation Causing Congenital Hypothyroidism in Three Generations of a Family Is Associated with Abnormalities in the Urogenital Tract Ana Carvalho, Pia Hermanns, Joao Anselmo, Luisa Rodrigues, Isabel Sousa, Luisa Mota-Vieira, Carlos Pereira-Duarte, Joachim Pohlenz Hospital Divino Espirito Santo, Ponta Delgada, Portugal; Hospital Divino Espirito Santo, Ponta Delgada, Portugal; Hospital Divino Espirito Santo, Ponta Delgada, Portugal; Children Hospital, Johannes-Gutenberd University, Mainz, Germany Although thyroid dysgenesis is the most common cause of congenital hypothyroidism (CH), its molecular basis remains elusive. Indeed, in only a minority of the cases (2-3%) it was possible to identify an underlying genetic defect. We studied a family with six affected members spanning three generations. The index case, (a 57-year-old white Azorean man) was diagnosed to have CH at 7-month of age when he presented severe impairment of suckling, constipation and poor development. Treatment with levothyroxine corrected the symptoms and produced catch up growth. His progeny of 2 males and 3 females are affected by CH and all but one diagnosed at neonatal screening. Ultrasound demonstrated normally located thyroid glands of reduced volumes. Four of the six affected family members, including the index case, present urogenital malformations as incomplete horseshoe kidney, undescended testicles and hydrocele). No other somatic abnormalities were found. Screening candidate genes known to cause thyroid dysgenesis revealed a new heterozygous mutation (c.74C[gt]G) in [italic]PAX8 gene[/italic] in all affected individuals. It leads to a substitution of proline with arginine at codon 25 (P25R). Fluorescence microscopy showed that P25R is normally located in the nucleus. In transient transfection studies this mutation causes reduced transcriptional activation ability when using a [italic]luciferase[/italic] reporter construct under the control of the [italic]thyroglobulin[/italic] promoter. This diminished transactivation ability is due to a loss of DNA binding capability as shown in electropgoresis mobility shift assay. We conclude that this novel [italic]PAX8[/italic] mutation is responsible for a severe form of dominantly inherited CH. The mutation in this family is associated with abnormalities of the urogenital tract.[br][br]Nothing to Disclose: AC, PH, JA, LR, IS, LM-V, CP-D, JP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 354 70 670 SAT-589 PO30-01 Saturday 609 2012


610 ENDO12L_SAT-590 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Increasing Incidence and Severity of Thyroid Damage 15 Years after [sup]131[/sup]I-Metaiodobenzylguanidine Treatment in Children with Neuroblastoma Hanneke Margo van Santen, Sarah Clement, Berthe LF van Eck, Lieve A Tytgat, Paul ASP van Trotsenburg Emma Children[apos]s Hospital, Academic Medical Center, Amsterdam, Netherlands; Academic Medical Center, Amsterdam, Netherlands; Emma Children[apos]s Hospital, Academic Medical Center, Amsterdam, Netherlands [bold]Introduction: [/bold]Thyroid dysfunction after treatment with [sup]131[/sup]I-Metaiodobenzylguanidine (MIBG) in children with neuroblastoma (NBL) has been reported, despite the use of thyroid protection. The incidence of thyroid dysfunction increases over time. For this reason we performed a long-term follow-up study.[br][bold]Aim of the study: [/bold]To determine the long term effects of [sup]131[/sup]I-MIBG treatment on the function and anatomy of the thyroid gland.[br][bold]Methods[/bold]: Fourty-three NBL patients treated with [sup]131[/sup]I-MIBG were evaluated. Patients received thyroid protection with potassium-iodide (100 mg KI per day).Thyroid dysfunction was defined as having a thyrotropin elevation (TE) [ge] 4.5 mU/L at the last moment of evaluation. In all survivors (n=16), ultrasound investigation of the thyroid gland was performed.[br][bold]Results: [/bold]Of the 43 patients, 26 patients developed TE (60.5%), after a mean time of 26 months, of which 16 patients (37.2%) developed permanent hypothyroidism. The average number of [sup]131[/sup]I-MIBG treatments was 3.0 per patient (range 1-9). On 95 (20.0%) of 476 MIBG-images uptake of radio-iodine was visible in the thyroid gland. No significant difference in radionuclide uptake in the thyroid gland was found between patients with TE and patients without TE. At the last moment of follow-up, 16 patients survived with an average follow-up time of 15.1 years (range 10.7-19.8 years). Of these 16, 50% had TE. In 8 of these 16 survivors thyroid nodules were found by ultrasound. One patient was diagnosed with papillary thyroid carcinoma 13 years after treatment with [sup]131[/sup]I-MIBG.[br][bold]Conclusion: [/bold]The occurrence of hypothyroidism and thyroid nodules after treatment with [sup]131[/sup]I-MIBG is high, and increases over time. Even thyroid carcinoma may occur. Protection of the thyroid gland with KI is insufficient. Considering that none of the patients showed clinical signs of thyroid dysfunction or palpable thyroid nodules, continuous screening for thyroid dysfunction and nodules in these survivors is necessary.[br][br]Nothing to Disclose: HMvS, SC, BLFvE, LAT, PASPvT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 165 70 671 SAT-590 PO30-01 Saturday 610 2012


611 ENDO12L_SAT-591 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Central Adrenal Insufficiency Could Not Be Confirmed by Measurement of Serum Basal DHEAS Levels in Pubertal Children Elisa Vaiani, Eduardo Chaler, Mercedes Maceiras, Juan Manuel Lazzati, Gisella Viterbo, Mariana Aziz, Silvia Gil, Alicia Belgorosky Hospital de Pediatria Garrahan, Buenos Aires, Argentina Central adrenal insufficiency (CAI) diagnosis remains challenging, particularly in partial deficiencies. Even though it has been proposed that measurements of baseline serum dehydroepiandrosterone sulfate (DHEAS) levels, mainly in adults, might be valuable in patients suspected of having CAI, many patients require dynamic biochemical studies, such as low-dose 1-[mu]g cosyntropin test, to assess adrenal function (LDT). In order to evaluate the usefulness of serum basal (B) DHEAS levels to confirm the diagnosis of CAI in adolescents, serum B SDHEA concentration as a function of serum cortisol (F) response to LDT was analyzed. Fifty six (26 females and 30 males) adolescent patients (mean[plusmn]SD chronological age: 14.2[plusmn]2.6 years) were studied. Patient diagnoses were the following: chronic corticoid treatment (CCT) (therapy had been withdrawn at least 6 month before, n=11), Multiple Pituitary Insufficiency (MPI), n=29), and others (n=16). All patients underwent LDT. A normal response was set at a serum F level [gt]496 nmol/L. Serum B SDHEA levels were matched for age and gender. According to our reference values, levels higher/equal to the 2.5[sup]th[/sup] percentile (per) of serum B DHEAS were considered normal. The salivary F (sF) level, as a marker of bio-available F, measured in the LDT was also used to confirm a normal response. Cut off sF level was established at 19.6 nmol/L. [bold]Results[/bold]: Normal LDT F response was found in 30 patients (Gr1) and below the cut off limit in 26 (Gr2) (Mean[plusmn]SD serum F levels: 584[plusmn]600 and 349[plusmn]100 nmol/L, respectively). In 8 patients of Gr1 (26%), serum B DHEAS levels were below, and in 14 patients of Gr2 (53.8%) above the 2.5[sup]th[/sup] per. Gr2 was divided in 2 subGrs according to the LDT F response (serum F levels below and equal/higher than 379 nmol/L, Gr2a (n=14) and Gr2b (n=12), respect.). Serum B SDHEA was normal in 6 patients (42.8 %) of Gr2a and in 8 patients (66.6%) of Gr2b. CAI was also confirmed by sF response to LDT in all patients with normal serum B DHEAS. [bold]Conclusion[/bold]: B DHEAS levels in pubertal children do not seem to be useful either to diagnose sufficiency or insufficiency central adrenal function. It could be speculated that as B DHEAS levels change dramatically during puberty in a relative short period of time, previous CCT or MPI might modified differently the dynamic changes of adrenal cortex zonation in these patients. Finally, dynamic testing is also necessary for assessing hypothalamic-pituitary-adrenal axis at puberty.[br][br]Sources of Research Support: FONCYT,Argentina.[br][br]Nothing to Disclose: EV, EC, MM, JML, GV, MA, SG, AB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 193 70 672 SAT-591 PO30-01 Saturday 611 2012


612 ENDO12L_SAT-592 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Growth and Clinical Features in Congenital Adrenal Insufficiency and Usefulness of ACTH Stimulation Test Jeesuk Yu, Ji Won Koh, Gu Hwan Kim, Han Wook Yoo Dankook University Hospital, Cheonan, Republic of Korea; Asan Medical Center, Seoul, Republic of Korea; Asan Medical Center, Seoul, Republic of Korea Objective: Congenital adrenal insufficiency caused by specific genetic mutation has various clinical manifestations including ambiguous genitalia, electrolyte imbance, hyperpigmentation and even life-threatening hyponatremic hypovolemic shock. Early suspicion and definite diagnosis is important. Neonatal screening test enables it, leading to better outcome, but sometimes ACTH stimulation test can be used as a diagnostic tool of adrenal insufficiency. The long term outcome still includes many morbid conditions such as obesity, short stature, precocious or delayed puberty and hyperandrogenism. This study is designed to evaluate the clinical manifestations including growth patterns and to find the usefulness of ACTH stimulation test.[br]Methods: Medical records including clinical and laboratory data were reviewed retrospectively. Medication dosage, genetic results and clinical features were compared according to specific disease types. Growth patterns were plotted based on chronologic age (CA) and bone age (BA). We defined [apos]BA advancement[apos] (BAA) when BA was advanced more than 6-12 months than CA.[br]Results: Sixteen patients with confirmed genotyping were enrolled in this study. Mutations of the gene 21-hydroxylase (CYP21A2, n=11), StAR (n=3), and DAX1 (n=2) were found. Six patients with gene CYP21A2 mutation were diagnosed by neonatal screening test. Growth evaluation showed BAA from 13 months of CA and it was markedly increased from 61 months and reached its peak at 73-84 months of CA. The hydrocortisone dosage was 21.13[plusmn]9.37 mg/m2 in BAA group compared with 21.42[plusmn]5.94mg/m2 in not BAA group (p=0.3). The average 17-OHP level was higher in BAA group, but not statistically significant. In ACTH stimulation test, stimulated 17-OHP levels were increased in all of the CYP21A2 mutation patients but not in the DAX1 gene mutation group. Obesity was found in 3 among 11 with CYP21A2 mutation. Mental retardation was found in 2 (100%) and precocious puberty in 1 (50%) among 2 patients with DAX1 mutation.[br]Conclusions: This study showed the importance of the revealing symptoms and neonatal screening test as well as gene analysis for early specific diagnosis and management. ACTH stimulation test would play an important role to support the diagnosis. Congenital adrenal insufficiency still has many morbid conditions such as advanced BA, obesity, and pubertal problems, so it will be required to adjust hydrocortisone dosage carefully and monitor growth and puberty.[br][br]Nothing to Disclose: JY, JWK, GHK, HWY 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 654 70 673 SAT-592 PO30-01 Saturday 612 2012


613 ENDO12L_SAT-593 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Cortisol Response to Synacthen Stimulation Is Attenuated Following Abusive Head Trauma Natasha Heather, Jose Derriak, Christine Brennan, Paul Hofman, Craig Jefferies, Wayne Cutfield University of Auckland, Auckland, New Zealand; Starship Children[apos]s Hospital, Auckland, New Zealand Background: Early life environmental stressors can lead to long-term alterations in the hypothalamic-pituitary-adrenal (HPA) axis. Abusive head trauma (the [apos]shaken baby syndrome[apos]) is a well recognised consequence of violence during infancy. However, the impact of abusive head trauma on the HPA axis is unknown. We sought to compare synacthen-stimulated cortisol responses in children who had suffered inflicted or accidental traumatic brain injury (TBI).[br]Methods: Children with a history of early childhood TBI were recruited from the Starship Children[apos]s Hospital databases (Auckland, New Zealand, 1992-2010). All were admitted to Starship Hospital aged less than 5 years with structural TBI (defined as a skull fracture, intra-cranial haemorrhage or cerebral lesion). Morning concentrations of serum cortisol, free thyroxine, free triiodothyronine, and thyroid stimulating hormone were measured. A low dose (1 mcg IV) synacthen test was performed, and serum cortisol measured at +30 and +60 minutes. In order to examine possible hormonal differences between inflicted (TBI[sub]I[/sub]) and accidental (TBI[sub]A[/sub]) groups, data were analysed using linear mixed models controlling for the appropriate confounding factors.[br]Results: We assessed 64 children with TBI[sub]I[/sub] and 134 with TBI[sub]A[/sub]. There were no differences in thyroid hormones between TBI[sub]I[/sub] and TBI[sub]A[/sub] subjects. Baseline cortisol concentration was also not different between groups, but TBI[sub]I[/sub] children displayed a 14% reduction in peak stimulated cortisol in comparison to the TBI[sub]A[/sub] group (p[lt]0.001), as well as reduced cortisol responses at +30 (p[lt]0.01) and + 60 minutes (p[lt]0.001). Importantly, cortisol concentrations were not associated with the severity of anatomical TBI (graded according to the Abbreviated Injury Scale). 52% of the TBI[sub]I[/sub] group were in foster care at the time of assessment and 40% of the TBI[sub]I[/sub] group reported interparental violence during pregnancy, so that the odds ratio of TBI[sub]I[/sub] children having a mother who suffered domestic violence during pregnancy was 6.2 times that of the TBI[sub]A [/sub]group (p[lt]0.001). However, we observed no cortisol difference associated with either violence during pregnancy or foster placement.[br]Conclusion: Synacthen-stimulated cortisol response is attenuated following inflicted traumatic brain injury in early childhood. We speculate that this is a consequence of chronic exposure to environmental stressors, as opposed to pituitary injury or early life programming.[br][br]Nothing to Disclose: NH, JD, CB, PH, CJ, WC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 599 70 674 SAT-593 PO30-01 Saturday 613 2012


614 ENDO12L_SAT-594 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) High Carrier Frequency of 21-Hydroxylase Deficiency Determined by Genotyping in Hellenic Population Antonia Dastamani, Maria Dracopoulou-Vabouli, Christina Merakou, Amalia Sertedaki, Ioannis Hatzis, George Antonakis, Georgia Dastamani, Elpidophoros Douratsos, George Lialios, Stavros Sifakis, Ilias Katsikis, Theodoros Stefos, Ioannis Triantafillidis, George P Chrousos, Catherine Dacou-Voutetakis Athens University, School of Medicine, Athens, Greece [bold]Backround:[/bold] The CYP21A2 genotyping is currently the most accurate method to define carrier frequency of CYP21A2 gene defects. The aim of the study was to define the by genotyping the carrier frequency of classical (C) and nonclassical (NC) 21-hydroxylase deficiency in a representative sample of the Hellenic population.[br][bold]Materials and Methods[/bold]: 497 neonates of Hellenic origin born in maternity hospitals located in various parts of Greece were studied. Cord blood DNA samples were used for genotyping of the CYP21A2 gene by Allele Specific PCR, searching for 14 molecular defects and direct sequencing whenever indicated. The 14 mutations account for 97% of the CYP21A2 defects previously found in CAH patients of our population. In order to detect possible gene duplications, Multiplex ligation-dependent probe amplification (MLPA) assay was employed in samples in which the severe classical p.Q318X mutation was identified.[br][bold]Results[/bold]: Out of 994 independent alleles studied, 53 mutant alleles were detected; 41 subjects were heterozygotes for either C (4) or NC (37) mutation, while 12 alleles carrying the p.Q318X mutation were associated with a duplicated CYP21A2 gene and consequently were neutral. The total heterozygote frequency detected was 8.25%. Heterozygote frequency for classical mutations was 0.8%, for non-classical mutations 7.44% and for CYP21A2 gene duplication with p.Q318X mutation 2.41%.[br][bold]Conclusions: [/bold]The observed heterozygote frequency in the Hellenic population, based on genotyping, is relatively high. The high frequency of CYP21A2 gene duplications associated with the p.Q318X mutation can be a source of error in diagnosing carrier status. The data strongly suggest that a precise identification of carriers, a prerequisite for neonatal screening policies and genetic counseling, should apply on molecular techniques and specific methodology for identifying gene duplication whenever the p.Q318X mutation is present.[br][br]Nothing to Disclose: AD, MD-V, CM, AS, IH, GA, GD, ED, GL, SS, IK, TS, IT, GPC, CD-V 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 264 70 675 SAT-594 PO30-01 Saturday 614 2012


615 ENDO12L_SAT-595 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Chemotype (Urinary Steroid Concentration)[ndash]Phenotype[ndash]Genotype Correlations in 53 Japanese Patients with 21-Hydroxylase Deficiency Yuhei Koyama, Keiko Homma, Masayuki Miwa, Kazushige Ikeda, Mitsuru Murata, Tomonobu Hasegawa Keio University School of Medicine, Tokyo, Japan; Mitsubishi Chemical Medience Co, Tokyo, Japan; Keio University Hospital, Tokyo, Japan; Keio University School of Medicine, Tokyo, Japan [bold][Introduction][/bold] The measurement of urinary steroid by gas chromatography/mass spectrometry (GC/MS) is useful for biochemical diagnosis of 21-hydroxylase deficiency (21OHD). Any data have not been reported regarding urinary steroid concentration (chemotype)-phenotype and chemotype-[italic]CYP21A2[/italic] genotype correlations in 21OHD. The aim of this study was to investigate chemotype, expressed as urinary steroid by GC/MS, -phenotype and chemotype-genotype correlations in 21OHD.[br][bold][Subjects and Methods][/bold] Fifty three patients with 21OHD (0-45 days of age) and 1890 control subjects (0-45 days of age) participated in this study. All subjects were Japanese. Chemotype: We measured urinary steroid metabolites of 17OHP, cortisol, aldosterone, and 21-deoxycortisol such as pregnanetriol (PT), tetrahydrocortisone (THE), tetrahydroaldosterone (THAldo), and pregnanetriolone (Ptl), using GC/MS (mg/g creatinine) and calculated SD value in each patient. Phenotype: Phenotype was classified as previously described such as salt wasting (SW, N=37), simple virilizing (SV, N=9), and non-classical (NC, N=7). Genotype: Genotype was determined by the sum of residual 21-hydroxylase activities (%) according to Finkielstain et al.[sup]1)[/sup] such as Null (0%, N=18), Severe ([gt]0-[le]1%, N=11), Moderate ([gt]1-[le]2%, N=12), Mild ([gt]2-[le]20-50%, N=9), and Unknown (N=3). We compared chemotype among three phenotypes (SW vs. SV vs. NC) and four genotypes (Null vs. Severe vs. Moderate vs. Mild). We also compared chemotype between classic and non-classic phenotype (SW+SV vs. NC), and sum of residual activities [le]2% and [gt]2% genotype (Null+Severe+Moderate vs. Mild).[br][bold][Results][/bold] Chemotype-phenotype: THAldo in SW was significantly lower than SV and NC. THAldo and Ptl showed significant difference between classic and non-classic phenotype. THE [lt]-2SD, THAldo [lt]-1SD, PT [gt]+6SD, and Ptl [gt]+16SD were shown in classic phenotype. THAldo [lt]-1SD were only in SW. Chemotype-genotype: THAldo in Null and Severe was significantly lower than Moderate and Mild. THAldo and Ptl showed significant difference between sum of residual activities [le]2% and [gt]2%. THE [lt]-2SD, THAldo [lt]-1SD, and Ptl [gt]+13SD were only shown in sum of residual activities [le]2% genotype.[br][bold][Conclusion] [/bold]We demonstrated chemotype expressed as urinary steroid concentration - phenotype and chemotype and[italic] CYP21A2[/italic] genotype correlations in 21OHD.[br][br](1)Finkielstain et al., J Clin Endocrinol Metab 2011; 96:E161.[br][br]Disclosures: YK: Employee, Mitsubishi Chemical Medience Co. Nothing to Disclose: KH, MM, KI, MM, TH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 530 70 676 SAT-595 PO30-01 Saturday 615 2012


616 ENDO12L_SAT-596 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Gender-Related Differences in Glucocorticoid Therapy and Growth Outcomes among Pubertal Children with 21-Hydroxylase Deficient Congenital Adrenal Hyperplasia (CAH) Jessica R Deslauriers, Anne M Lenz, Allen W Root, Frank I Diamond, Barry B Bercu Morsani College of Medicine at the University of South Florida, Tampa, FL; All Children[apos]s Hospital, St Petersburg, FL [bold]Context:[/bold] 21-Hydroxylase deficient CAH causes glucocorticoid and mineralocorticoid deficiency, hyperandrogenism, and short stature. Management of the pubertal patient with CAH is particularly challenging, as teenagers experience a critical growth spurt, the gonads produce androgens along with adrenal glands and noncompliance typically increases.[br][bold]Objective:[/bold] The purpose of this study was to determine if pubertal males and females with simple virilizing CAH (SVCAH) required different glucocorticoid dosages at progressive Tanner stages. The relationship between hydrocortisone dose and height during puberty and adult stature relative to targeted height was also assessed.[br][bold]Design and Setting:[/bold] This was a retrospective chart review study at an academic pediatric endocrinology unit.[br][bold]Patients:[/bold] Between January, 1981, and May, 2011, 867 patients were evaluated for precocious puberty. Twenty females and seventeen males with SVCAH were identified and followed through all stages of pubertal development.[br][bold]Main Outcome:[/bold] Males received a higher hydrocortisone dosage than females throughout all stages of pubertal development.[br][bold]Results:[/bold] The mean dose of hydrocortisone administered to males with SVCAH during puberty was 16.4[plusmn]4.8 mg/m[sup]2[/sup]/day of hydrocortisone; females received an average dose of 13.7[plusmn]4.6 mg/m[sup]2[/sup]/day. No significant gender-specific difference in glucocorticoid dosage was found at Tanner stage 2. However, the glucocorticoid dosage in males was significantly higher than in females at Tanner stages 3 through 5 (p [lsaquo] 0.05). Higher doses were associated with a shorter achieved than anticipated adult height. The mean adult height of patients with SVCAH was 9.6 cm shorter than their target height. Males had a less than expected anticipated adult stature than females, as males were on average 12.9 cm less and females 6.9 cm shorter than anticipated.[br][bold]Conclusions:[/bold] At Tanner stages 3-5, males with SVCAH required higher glucocorticoid dosages than did females. It is important to determine an optimal glucocorticoid dose for adolescents. A prospective study, carefully measuring compliance and ensuring follow-up, would be beneficial to determine the ideal hydrocortisone dose for adolescents with CAH.[br][br]Sources of Research Support: This project was funded by the Medicine and Gender Scholarly Concentration at the Morsani College of Medicine at the University of South Florida.[br][br]Nothing to Disclose: JRD, AML, AWR, FID, BBB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 977 70 677 SAT-596 PO30-01 Saturday 616 2012


617 ENDO12L_SAT-597 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Surgical Management of Congenital Adrenal Hyperplasia in Girls in France: A Population-Based Study Celine Castaings-Carlioz, Thomas Blanc, Veronique Tardy, Stephen Lortat-Jacob, Benedicte Coulm, Pierre Mouriquand, Joel Coste, Yves Morel, Alaa El Ghoneimi, Jean-Claude Carel H[ocirc]pital Robert Debr[eacute], Paris, France; H[ocirc]pital Robert Debr[eacute], Paris, France; Groupement Hospitalier Lyon Est, Lyon, France; H[ocirc]pital Necker Enfants Malades, Paris, France; Groupe Hospitalier Cochin-Saint Vincent de Paul, Paris, France; Groupement Hospitalier Lyon Est, Lyon, France; Groupe Hospitalier Cochin-Saint Vincent de Paul, Paris, France; Groupement Hospitalier Lyon Est, Lyon, France; H[ocirc]pital Robert Debr[eacute], Paris, France; H[ocirc]pital Robert Debr[eacute], Paris, France [bold]Background[/bold]: Despite the major long-term influence of feminizing surgery in girls with CAH and several consensus conferences, little is known of when and which type of surgery is performed at a population level.[br][bold]Patients and methods[/bold]: We analyzed the first surgical procedure performed in a population-based cohort of all 166 girls with a classical form of CAH due to 21-hydroxylase deficiency, born in France from 1996 to 2003.[br][bold]Results[/bold]: Patients who had surgery were classified according to Prader, after revision of the medical and surgical records by trained surgeons, as I (n=3), II (n=19), III (n=43), IV (n=49), V (n=6) and not available (n=22). Five of them had received a prenatal dexamethasone treatment. 142/166 (85,5%) had genital surgery at a mean age of 289,5 [plusmn] 427 days. Data concerning surgery were missing for 6 girls and the data are presented on 136 patients Pre-operative genitography was done in 99/136 girls (73%), and was informative to evaluate the level of uretro-vaginal confluence in 79. Endoscopy was performed in 64 girls, prior to surgery (n=16), or during feminizing genitoplasty (n=48). Among the 34 girls who had both genitography and endoscopy, the result was consistent in 30 (88%). Clitoridoplasty and vaginoplasty were performed during the same procedure in 118 patients (87%) whereas 4 patients had a two-stage procedure. Neurovascular sparing dismembered clitoridoplasty was performed in the majority of cases (n=76/121). Fortunoff flap with posterior colpotomy was the most frequent technique for vaginoplasty (n=69/123, 56%). The 142 patients were operated at 29 institutions throughout the country: 10 institutions operated a single patient while 5 institutions operated more than 8 patients during the 8-year period.[br][bold]Conclusion[/bold]: Our population-based study of the surgical management of CAH in France demonstrates a wide heterogeneity of surgical techniques performed by a large number of institutions throughout the country despite the rarity of the condition ([lt]20 cases/year). Similar studies should be undertaken in other countries to allow the establishment of evidence-based guidelines for the surgical management of CAH.[br][br]Nothing to Disclose: CC-C, TB, VT, SL-J, BC, PM, JC, YM, AEG, J-CC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2309 70 678 SAT-597 PO30-01 Saturday 617 2012


618 ENDO12L_SAT-598 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Obesity and Familial Predisposition Are the Main Determinant Factors of Adverse Metabolic Profile in Young Patients with Congenital Adrenal Hyperplasia Ricardo PP Moreira, Sandra MF Villares, Larissa G Gomes, Berenice Bilharinho Mendonca, Tania ASS Bachega Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil The introduction of glucocorticoid (GC) therapy allows normal life span for CAH patients; however, it is suggested an increased prevalence of obesity and metabolic syndrome (MetS), predisposing to adverse cardiovascular risk profile. There are limited data regarding the prevalence of MetS co-morbidities and their determinant risk factors in young CAH patients. [bold]Objective: [/bold]To assess the obesity and MetS prevalences in young CAH patients and to correlate with GC doses, treatment duration and family history. The impact of GC replacement on the prevalence of MetS components in CAH was analyzed through the comparison of metabolic profile between obese CAH patients and obese-matched controls. [bold]Patients and Methods:[/bold] 33 CAH young patients (21 SW, 12 SV) at a mean age of 11.9 [plusmn] 3.6 yrs receiving cortisone acetate (Mean 15.5 [plusmn] 3.1 mg/m[sup2]/day), salt wasters also receiving fludrocortisone (Mean 50 mcg/day). Treatment duration varied from 3.5-17.2 yrs and aimed to obtain normal androgen levels according to age/sex. Obesity was defined by BMI [gt]95[sup]th[/sup] for age/sex and MetS by NCEP/ATPIII-modified criteria. Familial history of obesity, dyslipidemia, hypertension, diabetes and cardiovascular disease was assessed. Data of 10 obese-matched controls were also evaluated. [bold]Results[/bold]: MetS and obesity were observed in 15% and 30% of CAH patients, respectively, which were higher than the reference population ([italic]P[/italic][lt]0.05). Low HDL-c levels were identified in 42% of CAH patients and increased blood pressure, waist circumference and triglycerides levels in 18%, 12% and 15% of patients, respectively. High fasting glucose level was identified in 1 patient. In CAH patients, BMI was positively correlated with higher systolic blood pressure, HOMA-IR and total cholesterol levels ([italic]P[/italic][lt]0.05), independently of GC dose, treatment duration, clinical form and sex. Positive family history of MetS components were higher in obese CAH as compared to non-obese CAH patients ([italic]P[/italic][lt]0.05). There was no difference in the lipid profile, blood pressure, HOMA-IR between obese CAH patients and obese-matched controls. [bold]Conclusions:[/bold] CAH young patients present higher prevalence of obesity and MetS and interestingly were not correlated with GC doses and duration of therapy. This study suggests that obesity and familial predisposition are the main determinant factors for and adverse metabolic profile in CAH.[br][br]Nothing to Disclose: RPPM, SMFV, LGG, BBM, TASSB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1206 70 679 SAT-598 PO30-01 Saturday 618 2012


619 ENDO12L_SAT-599 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Increased Abdominal Adiposity and Cardiovascular Disease (CVD) Risk in Adolescents with Congenital Adrenal Hyperplasia (CAH) Mimi Susan Kim, Anna Ryabets-Leinhard, Vicente Gilsanz, Mitchell Eugene Geffner Children[apos]s Hospital Los Angeles, Los Angeles, CA; Children[apos]s Hospital Los Angeles, Los Angeles, CA [bold]Aims:[/bold] In adolescents with classic CAH due to 21-hydroxylase deficiency, to (1) characterize CVD risk factors, including metabolic syndrome components, abdominal adiposity, and intimal-media thickness (IMT), and (2) describe the relationship among hyperandrogenism, adiposity, and insulin resistance.[br][bold]Methods:[/bold] Using a sex-, age-, ethnicity-, BMI-, and waist circumference-matched case-control design, we compared 18 adolescent cases with 18 controls. CT imaging was used to quantify visceral fat (VF), subcutaneous fat (SCF), and intrahepatic fat (IHF); DXA and anthropometry for body composition; Doppler carotid IMT to assess subclinical atherosclerosis; and fasting analytes (lipids, insulin, glucose, inflammatory markers, androgens, and SHBG). HOMA was calculated and blood pressure was evaluated.[br][bold]Results:[/bold] Compared to controls, adolescents with CAH (12-24 y, 61% female, 60% overweight/obese) had significantly increased VF (30.7 [plusmn]23.0 vs 18.7 [plusmn]19.1 cm[sup]2[/sup]; P [lt]0.01), and increased SCF compared to controls (228.7 [plusmn]163.9 cm[sup]2 [/sup]vs. 190.4 [plusmn]132.6 cm[sup]2[/sup]; P =0.058) Among cases, there was no gender difference in IMT or IHF. SHBG correlated negatively with abdominal adiposity on CT (r =-0.51; P [lt]0.05) and trunk fat on DXA (r =-0.59; P [lt]0.02), while androstenedione correlated positively with IMT (left-sided; r =0.50; P [lt]0.05). VF (r =0.52; P [lt]0.05) and SCF (r =0.57; 0.59; P [lt]0.05) correlated positively with insulin and HOMA, respectively. Both correlated positively with leptin (VF: r =0.66, P [lt]0.01; SCF: r =0.76, P [lt]0.001) and negatively with vitamin D (VF: r =-0.47, P =0.05; SCF: r=-0.57, P =0.02). Upper body fat assessed by DXA showed a positive association with HOMA (arms, trunk; r =0.51, 0.55; P [lt]0.05) and with leptin (arms, trunk, legs; r =0.85, 0.85; 0.87; P [lt]0.001).[br][bold]Discussion:[/bold] Compared to matched controls, adolescents with CAH show increased VF which correlated positively with fasting insulin, HOMA, and leptin. In addition, their upper body fat shows positive correlations with HOMA and leptin. These relationships may support a role for disordered regulation of leptin and insulin in CAH, and may predispose these patients to future CVD risk. Our additional findings that (1) lower levels of SHBG correlated with greater abdominal adiposity, (2) androstenedione was positively associated with IMT, and (3) there was a lack of sex difference in VF and SCF, IMT, and IHF in the CAH group suggest a pathological role for androgen excess or a lack of protective effect from estrogen.[br][br]Sources of Research Support: Saban Research Institute of Children[apos]s Hospital Los Angeles Clinical Research and Career Development Award and CTSI award (09-00261); Southern California CTSI at University of Southern California, Keck School of Medicine KL2 Mentoring Research Career Development Award (KL2RR031991).[br][br]Nothing to Disclose: MSK, AR-L, VG, MEG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1729 70 680 SAT-599 PO30-01 Saturday 619 2012


620 ENDO12L_SAT-600 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Arterial Hypertension in Children: Alterations in Mineralocorticoid and Glucocorticoid Axis and Their Impact on Pro-Inflammatory, Endothelial Damage, and Oxidative Stress Parameters Carmen Campino, Alejandro Martinez-Aguayo, Marlene Aglony, Hernan Garcia, Rodrigo Bancalari, Carolina Avalos, Lilian Bolte, Carolina Loureiro, Cristian A Carvajal, Lorena Garcia, Sergio Lavanderos, Carlos E Fardella Facultad de Medicina, Pontificia Universidad Cat[oacute]lica, Santiago, Chile; Facultad de Medicina, Pontificia Universidad Cat[oacute]lica, Santiago, Chile; Facultad de Ciencias Quimicas y Farmaceticas, Universidad de Chile, Santiago, Chile; Millennium Institute of Immunology and Immunotherapy, Santiago, Chile Arterial hypertension affects 4% of children but its pathogenesis has been poorly studied, as well as its impact at the level of cardiovascular damage and upon parameters which determine this injury. [bold]Objetive[/bold]: To evaluate in hypertensive children, the prevalence of alterations in mineralocorticoid and glucocorticoid axis and their impact on pro-inflammatory, endothelial damage and oxidative stress parameters. [bold]Patients and Methods: [/bold]We selected 306 children (5-16 year old) who were divided in three groups. Group 1: Hypertensives (n=111); Group 2: normotensives with at least one hypertensive parents (n=101); Group 3: normotensives children with both parents normotensives (n= 95). Fasting blood samples were drawn for measuring hormones (aldosterone, plasma renin activity (PRA), cortisol (F), cortisone (E)); inflammation variables (hsRCP, adiponectin, IL-6, IL-8, TNF-[alpha]); endothelial damage (PAI-I, MMP9 and MMP2 activities) and oxidative stress (malondialdehyde). Familial hyperaldosteronism type 1 (FH-1) was diagnosed when aldosterone/PRA ratio [gt]10 was associated with the presence of chimeric CYP11B1/CYP11B2 gene detected by XL-PCR. The 11[beta]-HSD2 activity was estimated by F/E ratio and it was considered altered when this ratio exceed the mean plus 2 SD respect to the group 3. The results were expressed as median [Q1-Q3] values. Comparison between the groups was done by Kruskal-Wallis and Dunn[apos]s Multiple Comparison Test. [bold]Results: [/bold]HF-1 was detected in 4/115 children (3.4%) in group 1 and none in the other 2 groups. The F/E ratio was elevated ([gt]4.3) in Group 1= 18/115 (15.6%); Group 2= 5/101 (4.9%) and Group 3= 5/95 (5.3%). The comparison between groups 1, 2 and 3 showed significant differences in levels of F (ug/dl): 9.9[6.7-14.4]*, 8.5[6.2-11.1], 8.4[6.3-10.4]; F/E ratio: 2.9[2.2-3.9]*, 2.8[2.2-3.3], 2.6[2.0-3.2]; hsCRP (mg/L): 1.2[0.4-2.3]*[sup],[/sup]**, 0.5[0.2-1.6], 0.5[0.2-1.3]; PAI-I (ng/ml): 22.2[13.4-31.7]*, 18.8[9.8-27.3], 14.9[8.9-23.3] and MMP-9 (number of changes respect to an internal control): 2.2[1.3-3.0]*, 1.8[1.2-2.5], 1.6[1.2-2.3].*p[lt]0.05 group 1 vs group 3, **p[lt]0.05 group 1 vs group 2. The others variables analyzed did not show significant differences between the groups. [bold]Conclusions: [/bold]In hypertensive children, HF-1 and 11[beta]-HSD2 deficit activity were detected. In addition, we found an increase in inflammation subclinical and endothelial damage variables. These results highlight the importance of routine blood pressure measurement in children population.[br][br]Sources of Research Support: This work was supported by Fondecyt 1100356, FONDEF D08I1087 and Nucleo Millenium on Immunology and Immunotherapy P07/088-F Chilean grants.[br][br]Nothing to Disclose: CC, AM-A, MA, HG, RB, CA, LB, CL, CAC, LG, SL, CEF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 285 70 681 SAT-600 PO30-01 Saturday 620 2012


621 ENDO12L_SAT-601 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Metformin Effect on Newborn Head Size, Maternal and Infant Weight Gain: A Follow-Up Study of an RCT on PCOS Sven Magnus Carlsen, Marit P Martinussen, Eszter Vanky Unit for Applied Clinical Research, Institute for Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway; St Olav[apos]s Hospital, Trondheim University Hospital, Trondheim, Norway; Norwegian University of Science and Technology, Trondheim, Norway BACKGROUND: The impact of metformin medication in pregnancy on maternal and offspring weight gain in PCOS is essentially unexplored. We aimed to investigate the impact of metformin on new-born anthropometry, and weight increase in mothers with PCOS and their infants.[bold][br]METHODS: [/bold]This is a follow-up study of a randomized controlled trial (The PregMet study), conducted in eleven secondary care centers. Women with PCOS were randomized to metformin (2000 mg daily) or placebo from first trimester to delivery. Questionnaires were sent to 256 participants one year postpartum; 199 responded. Maternal weight increase in pregnancy and one year postpartum and offspring anthropometry at birth and weight one year postpartum were registered. [bold][br]RESULTS:[/bold] Women randomized to metformin during pregnancy, gained more weight from the first trimester of pregnancy to one year postpartum, compared to those in the placebo group. Metformin- exposed girls had increased head circumference at birth compared to placebo-exposed ones in multivariate regression analyses (P =0.001). In boys, there was no difference between the groups. At one year of age, intrauterine metformin exposure was associated with increased body weight (10.2 [plusmn] 1.2 kg vs. 9.7 [plusmn]1.1 kg; p = 0.003) in the infants.[bold][br]CONCLUSIONS:[/bold] PCOS women randomized to metformin during pregnancy, had higher weight one year postpartum compared to those in the placebo group. Metformin exposure in utero seems to increase new-born head size in girls but not in boys at birth. At one year of age, metformin was associated with increased weight in both girls and boys.[br][br]Nothing to Disclose: SMC, MPM, EV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 394 70 682 SAT-601 PO30-01 Saturday 621 2012


622 ENDO12L_SAT-602 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Children Conceived with Ovarian Stimulation Have Shorter Stature Tim Savage, Mark Green, Harriet Miles, Fran Mouat, John Peek, Paul L Hofman, Wayne S Cutfield University of Auckland, Auckland, New Zealand; Fertility Associates Auckland, Auckland, New Zealand; The Liggins Institute, University of Auckland, Auckland, New Zealand [bold]Background[/bold]: Children conceived with the help of ovarian stimulation alone (OS[sub]A[/sub]), (not in vitro fertilization (IVF)) account for approximately 5% of all births in the developed world[sup]1[/sup]. Ovarian stimulation is part of the process of IVF, and studies have shown that children conceived with IVF are taller than naturally conceived children [sup]2[/sup]. We aimed to determine whether children conceived with ovarian stimulation alone would differ phenotypically and biochemically from naturally conceived children of fertile(time to conception [lt]12 months) and subfertile(time to conception [gt]12 months)parents[br][bold]Methods[/bold]: Healthy pre-pubertal children aged 3[ndash]10 years, born at term, after singleton pregnancies were recruited in Auckland (New Zealand), and were allocated into three groups: i) conception following OS[sub]A[/sub], and naturally conceived children of ii) subfertile and iii) fertile parents. Anthropometric, body composition by DEXA scan (total body fat %), endocrine (IGF-1, IGF-2, IGF-BP3), and fasting metabolic (cholesterol, LDL, HDL, triglycerides, glucose [amp] insulin) parameters were recorded. Children[apos]s heights and BMI were expressed as standard deviation score (SDS) and corrected for genetic potential (i.e. parental height or BMI)[br][bold]Results[/bold]: 352 children were studied: 84 OS[sub]A [/sub]subjects, 54 subfertile and 214 fertile controls. Children of subfertile and fertile parents did not differ in measured outcomes. Overall, OS[sub]A [/sub]children were shorter than subfertile (SDS score -0.08 [plusmn]0.09 [italic]vs[/italic] 0.32 [plusmn]0.07; [italic]P[/italic]=0.001) and fertile (SDS score -0.08 [plusmn]0.09 [italic]vs[/italic] 0.45 [plusmn]0.10; [italic]P[/italic]=0.004) control children when corrected for genetic height potential.[br]OS[sub]A[/sub] boys were shorter than both subfertile (SDS score -0.18 [plusmn]0.14 [italic]vs[/italic] 0.42 [plusmn]0.16; [italic]P[/italic]=0.03) and fertile (SDS score -0.18[plusmn]0.14 [italic]vs[/italic] 0.35 [plusmn] 0.08; [italic]P[/italic]=0.01) boys, and there was a trend towards OS[sub]A[/sub] girls being shorter than subfertile ([italic]P[/italic]=0.06) but not fertile girls ([italic]P[/italic]=0.17). OS[sub]A [/sub]children also had[sub] [/sub]lower corrected BMISDS than subfertile (SDS score -0.90 [plusmn]0.15 vs -0.37 [plusmn]0.17; [italic]P[/italic]=0.05) and fertile (-0.90 [plusmn]0.15 [italic]vs[/italic] -0.34[plusmn]0.10; [italic]P[/italic]=0.008) controls. Among endocrine and metabolic parameters measured, the only difference between groups was that fasting glucose was lower in OS[sub]A [/sub]children than in fertile controls (4.62 [plusmn]0.07[italic] vs [/italic]4.81 [plusmn]0.04; [italic]P[/italic]=0.006)[br][bold]Conclusions[/bold]: Children conceived by ovarian stimulation were shorter (2-3cms) than naturally conceived children of fertile and subfertile parents. We speculate that this height difference may be due to peri-conceptual imprinting by ovarian stimulation.[br][br]1. Schieve LA, Devine O, Boyle CA, Petrini JR, Warner L 2009 Estimation of the Contribution of Non[ndash]Assisted Reproductive Technology Ovulation Stimulation Fertility Treatments to US Singleton and Multiple Births. American journal of epidemiology 170:1396-407. 2.Savage T, Peek J, Hofman PL, Cutfield WS. Childhood outcomes of assisted reproductive technology. Hum Reprod. 2011 Sep; 26(9):2392-400.[br][br]Sources of Research Support: A Research Grant from the National Research Centre for Growth and Development (New Zealand).[br][br]Nothing to Disclose: TS, MG, HM, FM, JP, PLH, WSC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 515 70 683 SAT-602 PO30-01 Saturday 622 2012


623 ENDO12L_SAT-603 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Metabolic Outcomes in Children Born to Mothers with Severe Hyperemesis Gravidarum Ahila Ayyavoo, Paul Hofman, Jose Derraik, Sarah Mathai, Peter Stone, Frank Bloomfield, Wayne Cutfield University of Auckland, Auckland, New Zealand; National Research Centre for Growth and Development, Auckland, New Zealand; Auckland District Health Board, Auckland, New Zealand [bold]Introduction:[/bold][br]Hyperemesis gravidarum leads to alterations in maternal (and possibly fetal) nutrition throughout pregnancy. There are no data on long-term metabolic health outcomes of these offspring. We hypothesize that hyperemesis gravidarum leads to fetal nutritional compromise or physiological stress, which may programme later metabolism and cardiovascular status.[br][bold]Methods:[/bold][br]Two groups of healthy pre-pubertal children born at term, aged 4[ndash]11 years were studied: offspring of mothers who suffered hyperemesis gravidarum (HG group; n=37) and controls (n=55). Recruited HG children were born to mothers admitted to hospital with metabolic disturbance during pregnancy. Following an overnight fast, we performed a frequently sampled intravenous glucose tolerance test with insulin. Insulin sensitivity (Si) was calculated using Bergman[apos]s minimal model. Biochemical markers of metabolic syndrome were examined. Auxology and DEXA-derived body composition data were obtained. Data were analysed separately for boys and girls using linear mixed models, controlling for appropriate confounders. Data are expressed as mean[plusmn]SEM.[br][bold]Results:[/bold][br]HG boys (n=17) were taller (136.6[plusmn]1.7 vs 132.4[plusmn]1.5 cm; p=0.014), had lower Si (11.2[plusmn]0.8 vs 14.9[plusmn]1.1 x10[sup]-4[/sup] min[sup][minus]1[/sup][middot](mU/L); p=0.014), and higher fasting insulin (5.3[plusmn]0.5 vs 4.8[plusmn]0.5 mIU/L; p=0.047) in comparison to control boys (n=35). HG girls (n=20) compared to controls (n=18), also had higher fasting insulin levels (7.6[plusmn]0.9 vs 5.3[plusmn]0.7 mIU/l; p=0.037) and lower IGFBP1 (11.9 [plusmn]1.5 vs 17.4[plusmn]2.14 ng/ml; p=0.028). Conversely, height and Si were similar in HG and control girls, although baseline cortisol was higher in HG girls (262.4[plusmn]16.5 vs 200.6[plusmn]20.0 nmol/l; p=0.067). DEXA-derived body composition was similar in HG and control groups.[br][bold]Conclusion:[/bold][br]There seems to be sex-dependent metabolic and auxological differences in the offspring of mothers who suffered severe hyperemesis gravidarum. HG boys were less insulin sensitive and taller than controls. Long-term follow up of this cohort will be essential in determining later risk of metabolic disease.[br][br]Sources of Research Support: National Research Centre for Growth and Development, New Zealand; APEG Research Grants 2010 [amp] 2011.[br][br]Nothing to Disclose: AA, PH, JD, SM, PS, FB, WC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1732 70 684 SAT-603 PO30-01 Saturday 623 2012


624 ENDO12L_SAT-604 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Parental Origins Do Not Affect Adiposity, Food Preferences or Food Knowledge in Middle School Students Ranita E Kuryan, David Frankel, Barbie Cervoni, Audrey Koltun, Barbara Lowell, Lisa Altshuler, Michael Rosenbaum, Steven Shelov, Dennis E Carey, Phyllis W Speiser Cohen Children[apos]s Medical Center of NY/Hofstra University School of Medicine, New Hyde Park, NY; Cornell University, Ithaca, NY; Maimonides Medical Center, Brooklyn, NY; Columbia University Medical Center, New York, NY [bold][underline]Aim: [/underline][/bold][br]To compare measures of adiposity, food preference and food knowledge in middle school students with respect to parental origins and length of US residency.[br][bold][underline]Methodology:[/underline][/bold][br]The ROAD project is a 6 year, 5 school multicenter study, designed to assess clinical, behavioral, and biochemical risk factors for adiposity and adiposity related co-morbidities in a multiethnic urban middle school setting. Subjects were divided into groups based on the number of parents born in the US (0 or at least 1). Separate analyses for maternal and paternal country of origin (US, Asian, or other) were conducted. Subjects (N= 77, 62%F, mean age 12.0 [plusmn] 0.9) and their parents completed questionnaires regarding parental acculturation (country of origin, length of US residency) and family and personal medical histories. Student nutrition knowledge and dietary behaviors were assessed using revised Hearts N[apos]Parks[sup]1[/sup] subscales. Students also underwent anthropometric assessments (BMI and waist circumference). Z-scores were used to normalize data for age and gender. ANOVA and Pearson[apos]s correlation were used to compare adiposity measures, food knowledge and food preference scores between parental groups and length of residency, respectively.[br][bold][underline]Results: [/underline][/bold][br]There were no significant between-group differences in any of the variables. Specifically, there was no significant effect of number or gender of parents born outside of the US, or length of US residency, on measures of adiposity, food preference, or food knowledge.[br][bold][underline]Conclusion:[/underline][/bold][br]In adults there is a direct correlation between number of years lived in the US and Westernization of the diet [sup]2[/sup]. In toddlers there is a direct correlation of caregivers[apos] acculturation scores with consumption of sweets. In the same study, children cared for by Latinos consumed significantly less vegetables [sup]3[/sup]. In contrast, we found no significant effects of ethnicity or acculturation on anthropometry, nutrition knowledge, or dietary habits in middle school students. This may suggest a more rapid Westernization of dietary habits in early adolescence than has been noted previously. Further studies with larger populations are indicated.[br][br]1.http://www.nhlbi.nih.gov/health/prof/heart/obesity/hrt_n_pk/2002_report.pdf, accessed 12/8/11. 2. Sofianou A, Fung TT, Tucker K. Differences in diet pattern adherence by nativity and duration of the US residency in the Mexican- American population. J Am Diet Assoc. 2011; 111:1563-69.e2. 3. Erinosho TO, Berrigan D, Thompson FE, Moser RP, Nebelin LC, Yaroch AL. Dietary Intakes of Preschool-Aged Children in Relation to Caregivers[apos] Race/Ethnicity, Acculturation, and Demographic Characteristics: Results from the 2007 California Health Interview Survey [published online ahead of print December 10 2011]. Matern Child Health J. 2011. http://www.springerlink.com/content/55q04p5712222087. Accessed January 4 2012.[br][br]Sources of Research Support: Academy for Medical Development and Collaboration (AMDeC) by the Starr Foundation, support from NIH grant numbers UL1RR024156, UL1RR0023568 and CTSA grant # UL1RR024156 to Columbia University and NIH/NCRR GCRC grant #M01 RR018535 to NSLIJHS.[br][br]Nothing to Disclose: REK, DF, BC, AK, BL, LA, MR, SS, DEC, PWS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 184 70 685 SAT-604 PO30-01 Saturday 624 2012


625 ENDO12L_SAT-605 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Leptin, IGF-I/II during the First Weeks of Life Determine Body Composition at 2 Yrs in Infants Born with Very Low Birth Weight (VLBW) Maria Isabel Hernandez, Katherine Rossel, Veronica Pena, Gabriel Cavada, Alejandra Avila, German Iniguez, Varonica Mericq University of Chile, Santiago, Chile; Hospital Clinico San Borja Arriaran, Santiago, Chile; University of Chile, Santiago, Chile Rapid early growth is associated with adverse metabolic risks in young adults especially in small for gestational (SGA) age, independent if they were born at term or preterm. Aim: To determine whether there are differences in body composition (BC) between preterm born VLBW either appropriate for gestational age (AGA) or SGA and whether these differences are related to a certain period of first yr growth or metabolic variables during this period. 14 VLBWPT, (8 AGA, 6 SGA) followed as part of the premature infants follow-up program. BC by DEXA at 2 years corrected age, IGF-I/IGF-II and leptin weekly for 8 weeks and thereafter at 1, 3, 6 and 12 months. VLBWSGA children were lighter, but length and BC were not different. Total % fat mass correlated with Leptin and IGF-II concentrations obtained at 2 weeks (r2= 0.79, p=0.01 and r2=0.6, p= 0.05, respectively) and lean mass correlated inversely with IGF-II concentrations obtained at 2 weeks (r2= - 0.7, p=0.01) and Leptin at 8 weeks (r2= - 0.9, p=0.04). Bone mineral content correlated with IGF-I (r2=0.5, p=0.04,) and Leptin concentrations (r2=0.6, p=0.01) obtained at 6 weeks. Bone mineral density correlated inversely with IGF-II concentrations obtained at 1 and 4 weeks (r2 -0.7, r2 -0.5 p[lt]0.05, respectively) and directly with leptin concentration at 3 weeks (r2 0.58, p=0.02). Lighter VLBW infants have lower lean mass (p[lt]0.01). We did not find associations of BC with the different periods of first yr growth. In conclusion: body composition in VLBW SGA and AGA subjects was similar. Leptin, IGF-I, IGFII during the first 8 weeks of life determine BC at 2 yrs. The follow up of the complete sample for a longer period will show whether additional differences develop later.[br][br]Sources of Research Support: FONDECYT Grant 1090028.[br][br]Nothing to Disclose: MIH, KR, VP, GC, AA, GI, VM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 989 70 686 SAT-605 PO30-01 Saturday 625 2012


626 ENDO12L_SAT-606 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Serum Antioxidant Status Is Associated with Metabolic Syndrome among U.S. Adolescents: Findings from Recent National Surveys May A Beydoun, Jose A Canas, Hind A Beydoun, Xiaoli Chen, Monal R Shorff, Alan B Zonderman National Institutes of Health/Intramural Research Program, Baltimore, MD; Nemours Children[apos]s Clinic, Jacksonville, FL; Eastern Virginia Medical School, Norfolk, VA; Johns Hopkins University, Baltimore, MD; Michigan Public Health Institute, Okemos, MI BACKGROUND: Specific micronutrients were shown to have anti-inflammatory and antioxidant effects, including carotenoids, retinol, vitamin C and vitamin E, by reducing oxidative stress, a process that accompanies the pathogenesis of many chronic diseases. It is still largely unknown whether they are associated with the occurrence of metabolic syndrome (MetS) in the adolescent US population.[br]METHODS: MetS was defined by the International Diabetes Federation (IDF) criteria along with other outcomes namely elevated Homeostatic Model Assessment Insulin Resistance (HOMA-IR), C-reactive protein (CRP) and hyperuricemia. Retinol, retinyl esters, carotenoids [[italic][alpha][/italic]-carotene, [italic][beta][/italic]-carotene ([italic]cis+trans[/italic]), [italic][beta][/italic]-cryptoxanthin, lutein+zeaxanthin, total lycopene], vitamin E, and vitamin C were included as antioxidants. We tested associations between serum antioxidant status and MetS outcomes among adolescents aged 12-19y using cross-sectional data from NHANES 2001-2006 (n = 782 to 4,285).[br]RESULTS: IDF MetS prevalence was estimated at 7% among adolescent boys and 3% among girls. In adjusted models, adolescents with MetS had consistently lower serum carotenoid concentrations compared to their counterparts without MetS. Total carotenoids were also inversely related to HOMA-IR and CRP. Vitamin C was consistently inversely related to serum uric acid level and MetS binary outcome. Retinol+retinyl esters exhibited an inverse relationship with CRP and a positive relationship with uric acid and HOMA-IR as well as MetS binary outcome. Unlike other antioxidants, vitamin E had no consistent association with MetS and its components, particularly after controlling for serum cholesterol and triglycerides.[br]CONCLUSION: Among US adolescents, serum antioxidant such as carotenoids and vitamin C concentrations were inversely associated with MetS status, as well as HOMA-IR and CRP. Vitamin E had no consistent association with MetS and retinol had a positive relationship with HOMA-IR as well as MetS. These associations need further study.[br][br]Sources of Research Support: This study was entirely supported by the National Institute on Aging, Intramural Research Program (NIA/NIH/IRP).[br][br]Nothing to Disclose: MAB, JAC, HAB, XC, MRS, ABZ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 322 70 687 SAT-606 PO30-01 Saturday 626 2012


627 ENDO12L_SAT-607 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Prevalence of Obesity in an Ethnically Diverse Pediatric HIV Population Roy J Kim, Jennifer A Nwankwo, Janeen Graper, Theresa Barton University of Texas Southwestern Medical Center, Dallas, TX HIV infection disproportionately affects children and adolescents in minority groups. Minority children are also more likely to be obese and suffer from obesity related comorbidities. In spite of these factors, there remains a paucity of data with respect to the burden of obesity in the pediatric HIV population. We hypothesized that HIV infected children from minority groups would have a greater prevalence of obesity compared to non-minority HIV infected children, and similar to published national norms. Furthermore we hypothesized that HIV infected children from minority groups would have lower height and weight measurements compared to non-minority HIV infected children. We conducted a chart review of a racially diverse cohort of HIV infected children seen at an urban medical center and examined the prevalence of obesity and dyslipidemia. Age, height, weight, gender, markers of virologic control, and medications were recorded. Statistical analysis consisted of univariate and bivariate descriptive statistics stratifying by race and ethnic group. Obesity was defined as a body mass index (BMI) z-score [gt]=2.[br]The records of 180 patients (mean age 16, range 3-20y) were reviewed. The overall obesity prevalence was 7% compared to 18% from published population based norms. Within our cohort, obesity prevalence was 9.5% among non-Hispanic black individuals and 3% among Hispanic individuals however the differences were not statistically significant (Fisher exact =0.3). As continuous variables, height z-score, weight z-score, and BMI z-score did not differ significantly between race/ethnic categories. Underweight, defined as a weight z-score [lt] -2 was associated with Hispanic ethnicity (p=0.03). This relationship was maintained after controlling for markers of virologic control. For the entire group, height z-score correlated directly with CD4 nadir (Pearson correlation 0.17; p=0.03) as did the weight z-score (correlation 0.17, p=0.04). However, BMI z-score was not related to the CD4 nadir. Secular changes in the treatment of HIV over time may play a role in the association between virologic control and growth since age at the time of the data collection was associated with a lower CD4 nadir (p[lt]0.01), higher viral load (p=0.02), and lower height z-score (p=0.04).[br]In summary, obesity was less common among HIV infected pediatric cohort compared to non-infected population norms. BMI z-score or obesity status wasnot associated with race or ethnicity.[br][br]Sources of Research Support: RK is supported by NIH grant 7K23DK080644-02.[br][br]Nothing to Disclose: RJK, JAN, JG, TB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2361 70 688 SAT-607 PO30-01 Saturday 627 2012


628 ENDO12L_SAT-608 POSTER SESSION: Thyroid, Adrenal, Fetal Origins of Disease, Obesity (1:30 PM-3:30 PM) Comparison of the Prevalence of Cardiometabolic Risk Factors among Hispanic Adolescents with and without Polycystic Ovarian Syndrome Carley A Gomez-Meade, Gabriela Lopez-Mitnik, Sarah E Messiah, Aimee Mankodi, Adriana Carrillo University of Miami Miller School of Medicine, Miami, FL; University of Miami Miller School of Medicine, Miami, FL Polycystic ovary syndrome (PCOS) is a complex condition with substantial cardiometabolic consequences. Adult Hispanics with PCOS have higher cardiometabolic risk due to obesity, diabetes and hypertension compared to non-Hispanic whites (NHW)(1). Studies among NHW adolescents with PCOS have reported no increased cardiometabolic risk compared to non-PCOS, BMI-matched controls(2,3). Other studies have reported that rates of insulin resistance (IR) were similar among obese Hispanic adolescents with and without PCOS(4). The aim was to compare the prevalence of additional cardiometabolic risk factors (lipids, waist circumference [WC], blood pressure) in Hispanic adolescent with PCOS to obese Hispanic controls.[br]Utilizing a prospective cohort study design, 52 Hispanic adolescents with PCOS and 40 obese Hispanic controls (mean age 15.7yrs v 14.2yrs) recruited from the University of Miami pediatric endocrinology clinic in Miami, Florida. Body mass index (BMI), WC, systolic (SBP) and diastolic (DBP) blood pressure, low density lipoprotein (LDL), high density lipoprotein (HDL) and total (TC) cholesterol, triglycerides (TG), fasting insulin, and fasting glucose (FG) were obtained. HOMA-IR was calculated to assess IR (HOMA-IR=fasting glucose(mg/dL)xfasting insulin(mU/L)/405). Chi square analysis was used to assess differences in mean prevalence of cardiometabolic risk factors between the groups.[br]PCOS adolescents had significantly (1) lower BMI (35.1kg/m[sup]2[/sup] v 36.2kg/m[sup]2[/sup], p=0.05); (2) higher mean HDL levels (63.3mg/dL v 41.0mg/dL, p=0.02); but (3) higher DBP (76mmHg, 72mmHg, p=0.01) versus controls. While no significant differences were found between groups, mean elevated WC ([ge] 90[sup]th[/sup] %ile for age and sex) (68%, 85%, respectively), insulin levels (23.9[micro]U/mL, 24.9[micro]U/mL, respectively) and HOMA ([ge]3.1) (5.2 [53%], 5.1 [69%], respectively) were elevated in both PCOS and controls. Mean FG (80.5mg/dL, 83.3mg/dL, respectively), TC (163.9mg/dL, 155.6mg/dL, respectively) and SBP (122mmHg, 118mmHg, respectively) values were similar for PCOS and controls. PCOS adolescents had lower TG values compared to controls (97.6mg/dL v 113.7mg/dL). No FG were elevated ([gt]110mg/dL) for either group.[br]Our results show, in general PCOS and non-PCOS obese Hispanic girls share similar cardiometabolic risk. DBP was the only risk factor found to be significantly increased in the PCOS group. However, both groups showed elevated levels of WC, insulin and HOMA, all significant risk factors for type 2 diabetes.[br][br](1)Lo JC, Feigenbaum SL, Yang J, et al. Epidemiology and adverse cardiovascular risk profile of diagnosed polycystic ovary synrdome. J Clin Endocrinol Metab 2006;91(4):1357-63. (2)Glueck CJ, Morrison JA, Friedman LA, et al. Obesity, free testosterone, and cardiovascular risk factors in adolescents with polycystic ovary syndrome and regularly cycling adolescents. Metabolism 2006;55(4):508-14. (3)Rossi B, Sukalich S, Droz J, et al. Prevalence of metabolic syndrome and related characteristics in obese adolescent swith and without polycystic ovary syndrome. J Clin Endocrinol Metab 2008;93(12):4780-6. (4)Nur MM, Newman IM, Siqueira LM. Glucose metabolism in overweight Hispanic adolescents with and without polycystic ovary syndrome. Pediatrics 2009;124(3):e496-502.[br][br]Nothing to Disclose: CAG-M, GL-M, SEM, AM, AC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 979 70 689 SAT-608 PO30-01 Saturday 628 2012


629 ENDO12L_SAT-609 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Human Pancreas Agenesis and Cardiac Malformation Syndrome Due to a GATA6 Gene Defect Jean-Louis Blouin, Jeremy Bevillard, Anne Vannier, Corinne Gehrig, Michel Guipponi, Federico Santoni, Daniel Metzger, Valerie Schwitzgebel University of Geneva, Geneva, Switzerland; University of British Columbia, Vancouver, Canada; University of Geneva, Geneva, Switzerland Background: Pancreas agenesis is a rare monogenic disorder leading to neonatal diabetes and exocrine pancreatic insufficiency. Previously, mutations in two transcription factors, PDX1 and PTF1A, have been identified to cause human pancreas agenesis. These two proteins play a crucial role in early pancreas development in humans and mice. Over 40% of the human cases with pancreas agenesis remain however unsolved.[br]Clinical case: We diagnosed neonatal diabetes on day two of life in a boy with intrauterine growth retardation (birth weight 1766g at 38 weeks of gestation ([lt]10th percentile) and severe hyperglycemia, blood glucose level was 45mmol/l. Despite insulin replacement therapy, he showed failure to thrive resolving after exocrine pancreatic enzyme substitution. Abdominal ultrasound and CT-Scan did not identify any pancreatic tissue. Additionally the index case presented with a ventricular septal defect (VSD) and a transient neonatal jaundice.[br]Methods: We sequenced known (PDX1, PTF1A) and candidate genes important for pancreas and cardiac development (GATA4, ISL1), but did not identify any disease-causing mutations. Whole exome sequencing has recently become an excellent tool for the analysis of unresolved Mendelian diseases. We used in-liquid exome selection process (Sureselect XT HumanAllExon 50Mb) that targets the consensus coding sequence (CCDS) i.e. all known exons, as well as small non-coding RNAs from the human genome in our patient and his unaffected parents. With this approach and after different filtering steps of the variants, we identified a heterozygous de novo missense mutation in the GATA6 gene p.Arg456His which lies in the zinc finger domain of the GATA6 protein.[br]Conclusion: Based on animal models and a recent report by Lango Allen et al. (1), heterozygous mutations in GATA6 are thought to lead to pancreas agenesis combined with cardiac outflow tract malformations. Here we contribute by describing a second case of pancreas agenesis and cardiac malformation syndrome due to a GATA6 mutation. In contrast to the published case, our patient showed no developmental delay. GATA6 is normally expressed in the human brain and could thus interfere with brain development. We suggest that this apparent genotype-phenotype mismatch may be explained by a gene dosage effect or by the presence of another constitutive or somatic mutation in different target tissues.[br][br](1) Lango Allen, H. et al., GATA6 haploinsufficiency causes pancreas agenesis in humans. Nature Genetics. 2011;44;1; 20-22.[br][br]Nothing to Disclose: J-LB, JB, AV, CG, MG, FS, DM, VS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1709 71 690 SAT-609 PO54-01 Saturday 629 2012


630 ENDO12L_SAT-610 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) A Novel Homozygous Mutation in [italic]RFX6[/italic] in an Infant with Neonatal Diabetes, Gallbladder Agenesis, Duodenal Atresia and Intestinal Malrotation Jennifer P Concepcion, Mark W Daniels, Christina S Reh, Honggang Ye, Siri Atma W Greeley Children[apos]s Hospital Orange County, Orange, CA; University of Chicago, Chicago, IL [bold]Background:[/bold] Monogenic neonatal diabetes occurs due to mutations in over 20 genes. [italic]RFX6[/italic] encodes a transcription factor expressed in early gut endoderm and endocrine progenitors but is later restricted to mature pancreatic islets where it is highly expressed and driven by the master endocrine regulator [italic]NEUROG3[/italic]. Recently, bi-allelic [italic]RFX6[/italic] mutations were described as a rare cause of neonatal diabetes in seven cases with pancreatic and biliary hypoplasia and duodenal or jejunal atresia, termed the Mitchell-Riley syndrome.[br][bold]Clinical Case[/bold]: A male infant diagnosed prenatally with IUGR and duodenal atresia was born at 34 weeks with a low birth weight of 1375 gm to Vietnamese parents with no known consanguinity or family history of diabetes. Within 24 hrs after birth he developed severe hyperglycemia (446 mg/dL) and exhibited ready sensitivity to an insulin drip. In addition to duodenal atresia, laparotomy on day of life 5 confirmed gallbladder agenesis, annular pancreas, and intestinal malrotation with perforation. Low fecal elastase [lt]100 mcg/g stool (normal [gt]200 mcg/g) suggested the possibility of pancreatic exocrine insufficiency. His hospital course was complicated by short bowel syndrome, feeding intolerance, cholestatic jaundice and subsequent liver failure. At age 4 months he was listed for liver transplant, but died due to sepsis.[br]Genomic DNA was isolated from saliva after IRB approved informed consent was obtained. An [italic]RFX6[/italic] missense variant detected by whole exome sequencing was confirmed by bidirectional sequencing the [italic]RFX6[/italic] gene. Analysis using [italic]Mutation Surveyor[/italic] software revealed the novel homozygous K260T mutation in [italic]RFX6[/italic], with both parents being heterozygous carriers. The positively charged lysine is highly conserved at this position across multiple species. Change to the non-charged threonine was predicted to be damaging by the PolyPhen-2 HumDiv prediction model, which tends to be more informative for rare variants. The mutation is downstream of the DNA-binding domain that characterizes all RFX proteins, but may interfere with the transcriptional activity of the B domain.[br][bold]Conclusions: [/bold]Monogenic neonatal diabetes is heterogeneous but genetic testing based on consideration of phenotypic features can reveal a cause in the majority of cases, even when consanguinity is not known in rare recessive cases. Future study of human mutations will provide insight into the function of transcription factors such as [italic]RFX6[/italic] that are essential for endocrine development.[br][br]Sources of Research Support: Acknowledgements: US National Institutes of Health Diabetes Research and Training Center P60. DK020595, the Lewis-Sebring Family Foundation, and the Kovler Family Foundation. We thank the family for participation in this study.[br][br]Nothing to Disclose: JPC, MWD, CSR, HY, SAWG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1976 71 691 SAT-610 PO54-01 Saturday 630 2012


631 ENDO12L_SAT-611 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) A Case Report of Neonatal Diabetes Mellitus in Schinzel-Giedion Syndrome Adetokunbo Omoruyi, Suzanne Kingery, Michael B Foster, Kupper Wintergerst University of Louisville, Louisville, KY Background: Schinzel-Giedion Syndrome is a rare genetic syndrome first described in 1978. It is characterized mainly by mid face retraction, hypertrichosis, multiple skeletal anomalies, cardiac and renal malformations. No metabolic or biochemical abnormalities have yet been reported in these children. We present the first case of a patient with Schinzel-Giedion Syndrome developing neonatal diabetes mellitus.[br]Clinical Case: An African American neonate diagnosed clinically with Schinzel-Giedion Syndrome soon after birth. She was the product of a 37 week gestational pregnancy. She was small for gestational age with a birth weight of 1890g and length of 44.5cm. She fulfilled the clinical criteria for the diagnosis of Schinzel-Gideon syndrome as evidenced by dysmorphism, mid face hypoplasia, hydronephrosis, ambiguous genitalia, patent ductus arteriosus, patent foramen ovale, hypertrichosis, dysplastic nails and skeletal anomalies. Her karyotype was 46,XX. In the first month of life she had episodes of hypoglycemia which resolved with then subsequent development of hyperglycemia by 3 months of life. At the time of diagnosis, blood glucose levels while hospitalized were above 200 mg/dL. Basal insulin alone was initiated with subsequent noted increase in insulin requirements. Abdominal ultrasonography did not reveal structural anomaly of the pancreas. Genetic testing detected no known pathogenic sequence variants for genes KCNJ11, GCK and insulin promoting factor 1(IPF-1) all known to be associated with neonatal diabetes. However, a homozygous sequence variant of unknown significance for neonatal diabetes mellitus was reported in our patients IPF-1 sequence (Pro243dup). This patient did not have resolution of diabetes mellitus. At her last follow up appointment, she was 4 years 8 months old on a once daily dose of Detemir insulin. Her daily insulin requirement was about 0.3units/kg/day with a glycosylated hemoglobin level of 7.3%.[br]Conclusion: Although the biologic significance of the reported sequence variant found in this patients IPF-1 gene for neonatal diabetes mellitus remains unknown, this is the first case report of permanent neonatal diabetes mellitus in a neonate with Schinzel-Giedion Syndrome.[br][br]Nothing to Disclose: AO, SK, MBF, KW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1730 71 692 SAT-611 PO54-01 Saturday 631 2012


632 ENDO12L_SAT-612 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Role of Long-Acting Octreotide in Treating Hypoglycemia in 2 Cases with Beckwith-Wiedemann Syndrome Hiba Al-Zubeidi, Michael E Gottschalk, Ron S Newfield Rady Children[apos]s Hospital San Diego, San Diego, CA; University of California San Diego, San Diego, CA Background:[br]Hyperinsulinism associated with Beckwith-Wiedemann syndrome (BWS) can occur in [tilde]50% of cases, causing hypoglycemia of variable severity. Parenteral use of Octreotide may be indicated if unresponsive to diazoxide. There is limited data on use of depot octreotiode in BWS[br]Methods:[br]Chart review of 2 cases[br]Results:[br]Case 1: A female infant born LGA at 37 weeks gestation to a 31 year old mother. Pregnancy was uncomplicated except for fetal macrosomia. At birth had clinical features consistent with BWS: macroglossia, hemihypertrophy, small omphalocele and high [alpha] fetoprotein. On DOL1 developed hypoglycemia, requiring continuous glucose infusions. On frequent feeds, had severe hypoglycemia with relative hyperinsulinism: nadir glucose 10 mg/dl, insulin 2 uIU/mL, [beta]-hydroxybutarate 1.2 mg/dl, and noncontributory cortisol and growth hormone. She was started on diazoxide for persistent hypoglycemia on DOL10 and required very high doses, up to 22 mg/kg/d, but had persistent hypoglycemia, and developed pulmonary congestion. She was then switched to sc Octreotide TID, starting at 15 mcg/kg/d, titrated up to 40 mcg/kg/d. She was converted a week later to LAR depot-octreotide at 9 mg monthly and remains euglycemic at age 1 year.[br]Case 2:A female infant born LGA at 26 4/7 weeks, to a 32 year old mother. Neonatal course complicated by tracheostomy secondary to macroglossia, hypotension requiring hydrocortisone, congenital hypothyroidism and elevated [alpha] fetoprotein. Genetic testing showed variant BWS due to unbalanced translocation with partial trisomy 11 P and partial monsomy of chromosome 8. She developed hypoglycemia at 3 mo of life. She had relative hyperinsulinemia: glucose 10 mg/dl, insulin level 1uIU/ml, and [beta]-hydroxybutyrate 0.6 mg/dl, cortisol 2.3 mcg/dl. Remained hypoglycemic on hydrocortisone up to 20 mg/m2/d. She was partially responsive to diazoxide, so switched to sc octreotide, kept euglycemic on 19 mcg/kg/d given QID. Octreotide discontinued at 18 months old, and remained euglycemic on frequent feeds. At age 4 diazoxide she was restarted due to hypoglycemia while on frequent feeds, with good response. Due to extensive hypertrichosis from diazoxide she was switched to LAR octreotide three mo later, initially at 10 mg monthly. Dose raised to 10 mg Q 21 days with good response.[br]Conclusion:[br]Successful treatment with LAR octreotide can be achieved in infants with BWS who are either resistant or cannot tolerate diazoxide.[br][br]Nothing to Disclose: HA-Z, MEG, RSN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 313 71 693 SAT-612 PO54-01 Saturday 632 2012


633 ENDO12L_SAT-613 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Congenital Hyperinsulinemia with Grade 4 Intraventricular Hemorrhage (IVH): A Case Report Jahanara Begum-Hasan Carilion Clinic Children[apos]s Hospital, Roanoke, VA The signs and symptoms of neonatal hypoglycemia are well known; however, massive intraventricular hemorrhage associated with hypoglycemia has rarely been reported in the literature.[br]Here we report an 8-week-old baby girl, born at 36.5 weeks of gestational age to a 27-year-old Turkish mother. Her birth history was uncomplicated other than a precipitous delivery. She was noted to have low glucose (20 mg/dl) after birth, along with tachypnea. Her glucometer readings remained in the 20s to 30s in the first 20 hours of life despite peripheral IV glucose infusion (D10w initially then D12.5w) with the glucose infusion rate (GIR) at 6-10 mg/kg/min. Her glucose level improved initially for a short period but dropped again into the 20s and 40s. A central line (PICC) was then established with D25w. Her follow up glucometer readings were noted in 30s to 50s most of the time on days 3-7 with a brief period of normoglycemia despite an increased GIR of 16- 20 mg/kg/min.[br]The infant developed seizures on day 3 of life. A subsequent CT head showed extensive intraventricular and intraparenchymal hemorrhages which were reported as commonly seen only in earlier gestational age infants. The CT angiogram did not show any vascular abnormality.[br]The endocrine service consulted on day 3. With limited data available, the infant was diagnosed with Congenital Hyperinsulinemia (CH) based on the detectable serum insulin at the time of documented low glucose (26 mg/dl) requiring a very high GIR. The infant was started on Diazoxide and was able to be weaned off IVF as her glucose levels were normalized. Her other problems in the hospital included feeding difficulties which required Gastrostomy tube prior to discharge around 1.5 months later.[br]A genetic analysis was negative for common mutations for CH and her pancreas appeared normal on abdominal ultrasound. Follow up Cranial USs showed slow improvement of IVH with development of periventricular cystic encephalomalacia. A clinical follow up showed improved efficiency of oral feeding with stable glucose levels. The infant[apos]s development was also noted as being on target with stable head circumference.[br]In the absence of any other clear etiology, the massive brain hemorrhage in this case appears to be secondary to severe prolonged hypoglycemia. The management of hypoglycemia in a newborn may need to be readdressed to find a better approach to avoid prolonged hypoglycemia with increased consequences of morbidity and mortality.[br][br]Nothing to Disclose: JB-H 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2330 71 694 SAT-613 PO54-01 Saturday 633 2012


634 ENDO12L_SAT-614 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Factitious Hypoglycemia: A Manifestation of Youngster Bullying Mohammed NMN Ahmed, Abdulelah NMN Al Mutairdi, Talal NMN Al Otaibi King Faisal Specialist Hospital [amp] Research Center, Riyadh, Saudi Arabia Introduction:[br][ldquo]The greatest source of (youngster) stress is the one some inflicts on others: bullying.[rdquo] (Ruttimann). Bullying can result in social maladjustment w/grave outcome. We report 2 young ladies ages 15 (Pt.A) [amp] 19 yrs (pt.B) who took recourse to covert ingestion of glyburide (Gly) resulting in near fatal effects in one. A correct Dx was arrived upon referral to us. Both were provided w/psychological support.[br]Aim: To define factors predictive/diagnostic of factitious hypoglycemia.[br]Case Summaries:[br]Pt A:had a 2-yr hx of severe neuroglycopenic symptoms, intermittent loss of consciousness (LOC), seizures, misdiagnosed as epilepsy, [amp] intubated at outside hospital. She lost a year of schooling. An extensive evaluation done outside included: blood glucose (BG)2 mmol/l, [ldquo]hi INS[rdquo], but [ndash]ve yield for INS/receptor abys, total body CT/MRI, EEG. She continued to receive dextrose 10-25% infusions thru in-dwelling central venous catheter upon exhaustion of all peripheral access that resulted in catheter infection [amp] systemic sepsis w/admission to ICU outside for a month [amp] was then referred to us. Our labs: within 2-hrs. fast, BG 2 mmol/l, INS 137 pmol/l;(RR: 17.8-173/for nl BG),CP(0.98 nmol/l;RR: 0.38-0.65),PRO (32 pmol/l;RR:3-20), w/INS/CP ratio:0.14,PRO/CP: 0.03, +ve serum GLY.T he parents were [ldquo]shocked[rdquo] w/the revelation. W/discrete persuasion pt. admitted surreptitious GLY use due to bullying, had used 36 labs (180 mg) GLY on day of her outside ICU admission. A [ldquo]friend[rdquo] at school had established a network providing GLY to pt. [amp] several other girls. School authorities were contacted for corrective measures.[br]Pt. B: university student had neuroglycopenic symptoms, LOC [amp] recurrent hypoglycemia at her local hospital where she was hospitalized for 21 days w/IV glucose support. She was readmitted to same local hospital w/recurrence. Our evaluation: Within 6 hrs fast: BG 2.1, serum INS 403, CP 3.31, PRO 30, INS/CP ratio 0.12, PRO/CP 0.015, serum +v for GLY. pt. admitted to social conflict [amp] rejection by a life-long girl friend, leading to depression that pushed her to consume her father[apos]s prescribed supply of glyburide.[br]Conclusion: Factors predictive of FH include young age [amp] social maladaptation. FH is probably more common than realized. Biochemical/hormonal findings may be indistinguishable form insulinoma. The Dx can be established by documentation of hyperinsuilemia, hi Prol, hi CP, molar ratio of INS [amp] PRO to CP [lt]1 w/concurrent hypoglycemia [amp] circulating GLY.[br][br]Nothing to Disclose: MNMNA, ANMNAM, TNMNAO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 328 71 695 SAT-614 PO54-01 Saturday 634 2012


635 ENDO12L_SAT-615 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Opposing Phenotypes (Maturity Onset Diabetes of the Young and Congenital Hyperinsulinism) in a Novel Glucokinase Promoter Mutation (-84C[gt]G) Max Feldt Children[apos]s Mercy Hospital, Kansas City, MO Background: Glucokinase (GCK) is a key regulator enzyme in the pancreatic beta-cell. Mutations in GCK deregulate glucose homeostasis resulting in either mature onset diabetes of the young (MODY2) or neonatal hyperinsulinism. Mutations in the GCK promoter region (-71G[gt]C) have been found to alter GCK gene expression.(1) However, promoter mutation (-84C[gt]G) has not been described in the clinical presentation of MODY2 nor in hyperinsulinism.[br]Case report: Patient 1, a 10 year old male, originally presented with fatigue, weight loss, glucosuria, and hyperglycemia [gt] 300 mg/dl. He had no history of glucocorticoids or other medications taken at time of diagnosis. Family history was significant for multiple maternal family members with diabetes mellitus diagnosed at young ages with no diabetic complications noted. ICA-512/IA-2, insulin, and GAD-65 antibodies were negative. Glargine, initiated at 0.1 U/kg with adjustments from 0.1-0.2 U/kg, improved glycemic control. HBA1c ranged from 5.2-6.1% during therapy. Commercial MODY gene analysis revealed a c.1a-84C[gt]G promoter region mutation of GCK. All other genetic results were negative. Glargine was discontinued. Twelve months later Patient 1[apos]s HBA1c was 5.6%.[br]Patient 2 is unrelated and is currently 3 years of age. She initially presented at 5 months of age with a hypoglycemic seizure. Blood glucose at the time of seizure was 37 mg/dl with undetectable beta-hydroxybutyrate. Extensive endocrine and metabolic evaluations were unremarkable. Presumed diagnosis was hyperinsulinism. She was initially started on cornstarch but persistent hypoglycemia necessitated initiation of diazoxide 5 mg/kg. Genetic screening for ABCC8 and KCNJ11 was negative. MODY gene analysis found the same c.1a-84C[gt]G GCK promoter mutation as Patient 1. Over the past few years, her dose of diazoxide was reduced to 2.9 mg/kg with good control of hypoglycemia.[br]Conclusion: A novel GCK promoter mutation has been identified in two patients with opposing phenotypes of MODY2 and hyperinsulinism. This emphasizes the need to include promoter regions as part of our arsenal of diagnostic genetic screening. Functional studies on these patients may help to determine if this promoter region mutation alters GCK gene expression or is a benign polymorphism.[br][br](1) Gasperikova D et al., Diabetes 2009; 58:1929-35.[br][br]Nothing to Disclose: MF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1346 71 696 SAT-615 PO54-01 Saturday 635 2012


636 ENDO12L_SAT-616 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Hyperinsulinism Due to an Activating Glucokinase Mutation Presenting as Severe Hypoglycemia in Late Adolescence Marielisa Rincon, Melissa Carlucci, Whitney Brown TC Thompson Children[apos]s Hospital, Chattanooga, TN; Palmetto Health Children[apos]s Hospital, Columbia, SC [bold]Background: [/bold]Congenital hyperinsulinism (CHI) is characterized by inappropriate oversecretion of insulin, typically presenting as hypoglycemia during the newborn period and infancy. Gain-of-function mutations of the glucokinase (GCK) gene represent a rare cause of CHI. We present a [italic]Val452Leu[/italic] mutation of the GCK gene in a patient with initial diagnosis of CHI in late adolescence.[br][bold]Clinical Case: [/bold]A 17 year old female was brought to the ED after a first episode of syncope. She was found to have blood glucose (BG) of 20 mg/dL by the emergency response services. She received several intravenous (IV) boluses of D50 and upon arrival to the ED her glucose was up to 60 mg/dL. A critical sample was not obtained prior to this intervention. She remained in her local hospital for 4 days on dextrose containing IV fluids, but was unable to maintain BG over 60 mg/dL. At this time she was transferred to our institution for further evaluation. She had a normal physical exam. Her past medical history was said to be unremarkable, but upon further investigation it was found that she had low BG as a neonate, but this resolved prior to her discharge from the nursery. She had also a history of seizures during her early school years, that resolved by age 12. BG were never documented in relationship to her seizures, however she stated that she learnt how to avoid a seizure by eating when she was somewhat lightheaded. Her family history was negative, with no known hypoglycemia in parents or siblings. Upon arrival, she was started on an observed fast. At 2 hours into the fast her BG dropped to 47 mg/dL. At the time, her insulin level was 15.6 uU/mL, c-peptide was 3.10 ng/mL, urine ketones were negative and she had normal ammonia and bicarbonate levels. After a glucagon challenge her glucose increased by 100 mg/dL. Counter regulatory hormone responses were normal and a sulfonylurea screen was negative. She was started on Diazoxide and octreotide, titrating to effect and eventually was able to be taken off IV fluids. Because of the above presentation, genetic testing was sought, demonstrating a [italic]Val452Leu [/italic]mutation of the GCK gene. Ultimately she was discharged home on Diazoxide, since pancreatectomy has not been particularly successful in similar cases. Her compliance with the medication has remained an issue.[br][bold]Conclusions: [/bold]GCK mutations must be considered in the differential diagnosis in any individual presenting with apparent new onset hyperinsulinemic hypoglycemia regardless of age.[br][br]Nothing to Disclose: MR, MC, WB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 166 71 697 SAT-616 PO54-01 Saturday 636 2012


637 ENDO12L_SAT-617 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Diabetic Ketoacidosis with Alkalemia Presenting with a Picture of Superior Mesenteric Artery Syndrome Heba Ismail, Marielisa Rincon Seattle Children[apos]s Hospital, Seattle, WA; TC Thompson Children[apos]s Hospital, Chattanooga, TN [bold]Background: [/bold]Diabetic ketoacidosis (DKA) with alkalemia is a rarely recognized metabolic alteration and presentation of diabetes. Vomiting, diuretic use, alkali ingestion and hypercortisolism have been implicated as causes. We describe a case of superior mesenteric artery syndrome (SMA) as the underlying cause of DKA with alkalemia.[br][bold]Results: [/bold]A 13 year old white male presented to the ED with the chief complaint of severe epigastric pain associated with non-bloody and non-bilious vomiting. Both symptoms developed about 36 hours prior to the presentation. His review of symptoms was significant for decreased appetite, documented weight loss of about 10 pounds over a 2 months period and polyuria and polydipsia for several weeks. Physical exam demonstrated dehydration, but otherwise was unremarkable. His initial laboratory data were remarkable for serum glucose of 689 mg/dl with a serum CO2 of 34 mmol/L (normal 21-33 mmol/L) and serum chloride of 88 mmol/L (normal 98-110 mmol/L). His body habitus, the history of polyuria and polydipsia, and the fasting serum glucose of [ge] 200 mg/dl met the diagnostic criteria for diabetes mellitus. He was initially managed with IV fluids and IV insulin and eventually transitioned to a subcutaneous insulin regimen, once clinically and metabolically stable.[br]It was initially thought that an acute gastroenteritis unmasked a latent type 1 diabetes mellitus. However, since the metabolic alkalosis was out of proportion to the short history of vomiting, an upper GI series was ordered. The images showed evidence of functional obstruction with dilatation in the descending portion of the duodenum together with sluggish peristalsis across the midline in the supine position. In the oblique position, contrast flowed across the midline easily. These findings were consistent with SMA syndrome. It was thought that this radiologic diagnosis of SMA syndrome was secondary to his recent weight loss triggered by diabetes, and that weight gain with adequately controlled diabetes would likely replace diminished abdominal fat and relieve the obstruction. The patient did well following discharge with no recurrence of symptoms.[br][bold]Conclusions: [/bold]We describe here a unique and interesting case of new onset diabetes presenting with metabolic alkalosis secondary to SMA syndrome. To our knowledge, this is the first case report of its kind.[br][br]Nothing to Disclose: HI, MR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 92 71 698 SAT-617 PO54-01 Saturday 637 2012


638 ENDO12L_SAT-618 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Treatment of Diabetes Mellitus in Rabson-Mendenhall Syndrome with U-500 Regular Insulin Administered by Continuous Subcutaneous Insulin Pump and Injections of Mecasermin Joel Steelman, Paul Thornton, John Dallas, Jill Radack, Michael Wilcutts Cook Children[apos]s Medical Center, Fort Worth, TX Background: Rabson-Mendenhall syndrome (RMS) is caused by a mutation in the insulin receptor gene. Severe insulin resistance, post-prandial hyperglycemia, and risk for fasting hypoglycemia due to impaired insulin clearance are seen with RMS. Children with RMS eventually develop diabetes mellitus and require very high insulin doses to achieve metabolic control.[br]U-500 regular insulin given either by multiple daily injections (MDI) or insulin pump (CSII) is an option for treatment of diabetes mellitus associated with extreme insulin resistance such as seen in RMS. The slower onset of action and longer duration are challenges for use of U-500 in RMS.[br]Therapies in combination with insulin such as metformin, thiazolidinediones, recombinant IGF1 (Mecasermin), and recombinant leptin have been tried in RMS.[br]Clinical Case: We report the use of regular U-500 insulin by CSII combined with twice daily Mecasermin injections in an 8 year old with RMS.[br]Diabetes mellitus without autoimmunity was diagnosed at 17 months old. Therapy was changed at age 6 from a split mix regimen to MDI using high doses of Glargine and Aspart due to deteriorating metabolic control. Poor metabolic control persisted on MDI.[br]The patient was hospitalized for severe diabetic ketoacidosis (DKA). Admission hemoglobin A1c was 12.7%. An insulin drip rate of 1.4 units/kg/hour for 36 hours was needed to correct DKA. An attempt to resume MDI was unsuccessful requiring resumption of insulin drip for 24 hours.[br]CSII with U-500 insulin was started. A lower basal rate was used overnight and meal boluses were given 30 minutes prior to eating. Final hospital CSII total basal rate was 38.9 units/kg/day with 67.7% of the total basal rate delivered over a 14 hour [quot]prandial period[quot]. Blood sugar means were 245 mg/dl for daytime and 110 mg/dl for fasting.[br]No improvement was seen using Pioglitazone and Metformin with CSII, and both drugs were stopped. Twice daily Mecasermin was started and improvement in metabolic control was seen. 67.7% of current dose of 253 mcg/kg/day is given in the morning. 95.7% of the current total daily CSII basal rate is given in a 12 hour [quot]prandial period[quot]. Current hemoglobin A1c is 11%.[br]Conclusions: U-500 insulin given by CSII is effective in our RMS patient. The addition of Mecasermin lowered insulin requirement and improved metabolic control. CSII provides benefit in RMS diabetes by allowing widely differing basal rates in prandial vs fasting periods thus minimizing glycemic excursion.[br][br]Disclosures: MW: Speaker Bureau Member, Ipsen; Scientific Board Member, Ipsen. Nothing to Disclose: JS, PT, JD, JR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 818 71 699 SAT-618 PO54-01 Saturday 638 2012


639 ENDO12L_SAT-619 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) C-Peptide[ndash]Positive, Autoantibody-Negative Diabetes Associated with a Confirmed 8p23 Deletion [mdash] Further Evidence for the 8p Region as a Locus for Autoantibody-Negative Diabetes Heba Ismail, Charles E Shaw-Smith, Craig E Taplin Seattle Children[apos]s Hospital, Seattle, WA; Royal Devon and Exeter NHS Foundation Trust, Exeter, UK Background: Previous reports have identified a susceptibility locus for both type 2 diabetes and monogenic diabetes (MODY) on chromosome 8p23. Candidate genes in this region include BLK and PPP1R3B.[br]Clinical Case: A 13 year old Hispanic girl presented with a 6 month history of polyuria, weight loss and fatigue. She had no family history of diabetes. Hyperglycemia was confirmed with a BGL of 360mg/dl and HbA1c was 12.6%. Urine showed glucosuria but no ketonuria, and she was not in DKA. Autoantibodies were negative for insulin, IA-2 and GAD, while C-peptide at presentation was 2.6 ng/mL (0.8-3.1). Subsequent ZnT8 autoantibodies were also negative. She was commenced on insulin.[br]From thereon her glycemic control has been excellent with HbA1c between 5.6% - 6.6% on approximately 0.2 units/kg of insulin glargine daily and a small amount of prandial rapid-acting insulin analogue. Her C-peptide measured 26 months after diagnosis (32 months from onset of polyuria) was 2.9 ng/mL, consistent with significant persistent endogenous insulin production. She is not obese and has no clinical evidence of insulin resistance. She has no evidence of diabetes complications to date.[br]At age 5, prior to the diagnosis of diabetes, she underwent chromosomal analysis due to global developmental delays that identified a chromosome 8p deletion. We recently performed a repeat DNA analysis by genome-wide SNP array which confirmed a deletion in the 8p23 region of approximately 11 Mb.[br]Conclusions: This case demonstrates a case of autoantibody negative diabetes presenting in childhood associated with persistent endogenous insulin production in a Hispanic child with a known deletion of chromosome 8p23. Previous studies have linked the 8p23 region to defects in insulin responses to glucose, particularly in Mexican American adults. The 8p region may include genes involved in insulin secretion and/or synthesis, and this case provides further evidence for this. To our knowledge, no cases of antibody negative diabetes in childhood have been reported associated with a known deletion of chromosome 8p23.[br][br]Borowiec M, et al: Mutations at the BLK locus linked to maturity onset diabetes of the young and [beta]-cell dysfunction. PNAS. 106; 34: 12260-14465, 2009. Cai G, et al: Genome-Wide Scans Reveal Quantitative Trait Loci on 8p and 13q Related to Insulin Action and Glucose Metabolism The San Antonio Family Heart Study. Diabetes; 53: 1369-1374, 2004.[br][br]Nothing to Disclose: HI, CES-S, CET 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 593 71 700 SAT-619 PO54-01 Saturday 639 2012


640 ENDO12L_SAT-620 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Type 1 Diabetes Mellitus in a Patient with Homozygous Sickle Cell Anemia Zohreh Shoar, Geoffrey Rezvani St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA Background: In children with sickle cell disease (SCD) reports of type 1 diabetes mellitus (T1DM) are extremely rare. Several studies fail to show the co-existence of the two conditions. In the few reported cases the diagnosis has been made based on the clinical presentation. Here we report a case of a patient with sickle cell anemia who presented with hyperglycemia and islet cell antibodies, a marker of T1DM.[br]Clinical Case: A 13 year old male with SCD was seen in endocrine clinic for hyperglycemia. Elevated finger stick blood glucose (BG) concentrations were detected while he had multiple random and fasting checks at the time of blood transfusions. He had history of elevated ferritin levels, splenectomy at age 5 years, and multiple transfusions since age 7. Physical examination showed no signs of hyperinsulinemia. Initial laboratory work up showed elevated Glutamic Acid Decarboxylase-65 antibodies of [gt]30 U/mL, IA-2 antibody of 1.1 U/mL, negative anti-insulin antibodies of [lt]0.4 U/mL, normal insulin of 3 uIU/mL, elevated BG of 129 mg/dL, ferritin of 2950 ng/mL, and HbA1C of 6.8%. He had normal kidney function and liver enzymes. Due to the short lifespan of red blood cells in hemoglobinopathies and the effect of storing of donor blood in dextrose before transfusions, HbA1C may not be reliable measure of glycemic control in patients such as ours. Therefore, we measured fructosamine to assess the average BG concentration during the past weeks; it was 327 uMOL/L (normal 190 to 270 uMOL/L). Since his BG levels were consistently elevated, treatment with subcutaneous insulin was initiated.[br]Conclusion: To our knowledge, this is the first patient reported with homozygous SCD, clinical picture consistent T1DM and anti-islet cell antibodies. In patients with iron overload, hemochromatosis can cause hyperglycemia, so we cannot rule out the possibility that this also contributed to our patient[apos]s hyperglycemia. Concurrent T1DM and SCD in children have been previously in 2 separate cases reported. However, none of those cases been proven to have positive pancreatic islet cell cytoplasmic antibodies. The lack of reported cases of T1DM in patients with homozygous SCD suggests that SCD may in some way be protective against the development of T1DM, but this report demonstrates that the two conditions can occur simultaneously.[br][br]Nothing to Disclose: ZS, GR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1149 71 701 SAT-620 PO54-01 Saturday 640 2012


641 ENDO12L_SAT-621 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Mutation in the [italic]CYP11A1[/italic] Gene Resulting in Partial Deficiency of P450 Side-Chain Cleavage Enzyme in a Patient with 46,XY Disorder of Sex Development (DSD) and Late-Onset Adrenal Insufficiency (AI) Charmian A Quigley, Saroj Nimkarn, Taninee Sahakitrungruang Indiana University School of Medicine, Indianapolis, IN; Weill Cornell Medical College, New York, NY; Chulalongkorn University, Bangkok, Thailand Formerly considered incompatible with fetal survival, P450scc (cholesterol side-chain cleavage enzyme) deficiency (D) is a very rare form of congenital adrenal hyperplasia that disrupts the 1st step of steroidogenesis, resulting in deficiencies of all adrenal and gonadal steroids. Early reported cases had 46,XY DSD and early-onset AI[sup]1[/sup][sup],2[/sup]. However, recently a handful of partial P450sccD cases due to mutations of [italic]CYP11A1 [/italic]have been reported in patients with late-onset (often life-threatening) AI, with or without DSD[sup]3,4[/sup].[br][bold]Objective[/bold]: to determine the cause of 46,XY DSD and late-onset AI in a 2-y-old child.[br][bold]Patient and Methods[/bold]: A phenotypically female Venezuelan child presented at birth with discordant prenatal karyotype (46,XY) and postnatal genital appearance. The baby had palpable left inguinal gonad, minimal clitoromegaly, and no hyperpigmentation. At age 1 month (m), she had normal gonadotropins, but low testosterone (30.3 ng/dL) and high DHEAS (131 [mu]g/dL; reference range [RR]: 0.5-19.4). Left orchidectomy at 3m revealed an immature but otherwise normal testis. At 1.3y she had normal serum anti-mullerian hormone and inhibin B for age. At age 1.5y, she presented to a local ER with acute gastroenteritis, lethargy, mild hyponatremia (given IV fluids); serum cortisol was 19.1 [mu]g/dL. Subsequent vomiting illnesses also resulted in ER treatment. At age 2.3y, she presented a different ER with vomiting, fever, mild hypotension and tachycardia: Na+ 122 mMol/L, CO2 16 mMol/L, glucose 49 mg/dL. Critical sample prior to IV hydrocortisone showed ACTH 250 pg/mL, cortisol 12 [mu]g/dL (low in context of physiological stress), plasma renin activity 11.15 ng/mL/hr (RR: 0.25-5.82), and low (but measurable) steroids of the other pathways. She was started on hydrocortisone and fludrocortisone with a provisional diagnosis of high-level steroidogenic defect.[br][bold]Results[/bold]: [italic]StAR[/italic] and [italic]HSD3B2[/italic] gene sequences were normal. [italic]CYP11A1[/italic] had homozygous c.1076C[gt]T substitution in exon 6, changing A359[gt]V; parents were heterozygous carriers. Prior [italic]in vitro[/italic] studies showed [sim]11-36% of native enzyme activity[sup]4,[/sup][sup]5[/sup], consistent with the clinical diagnosis of partial P450sccD.[br][bold]Conclusions[/bold]: This child[apos]s case demonstrates a broader clinical spectrum of P450sccD than initially thought; partial loss-of-function [italic]CYP11A1 [/italic]mutation should be considered as a differential diagnosis of 46,XY DSD, particularly in context of any clinical features of AI, such as hyponatremia/hypotension during illness, which may be life-threatening.[br][br]1. Tajima T et al. J Clin Endocrinol Metab 2001;86:3820-5. 2. Kim CJ et al. J Clin Endocrinol Metab 2008;93:696-702. 3. Sahakitrungruang T et al. J Clin Endocrinol Metab 2011;96:792-8. 4. Parajes S et al. J Clin Endocrinol Metab 2011;96:E1798-806. 5. al Kandari H et al. J Clin Endocrinol Metab 2006;91:2821-6.[br][br]Nothing to Disclose: CAQ, SN, TS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 260 71 702 SAT-621 PO54-01 Saturday 641 2012


642 ENDO12L_SAT-622 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Two Novel [italic]HSD3B2[/italic] Missense Mutations with Different Enzymatic Activity for [Delta]5 Steroids: Possible Cause for Elevated Levels of 17-Hydroxyprogesterone (17-OHP) in 3[beta]-Hydroxysteroid Dehydrogenase (3[beta]-HSD) Deficiency Patients Kei Takasawa, Kenichi Kashimada, Makoto Ono, Yohei Matsubara, Atsushi Hijikata, Noriyuki Katsumata, Masanori Takagi, Osamu Ohara, Tomohiro Morio, Shuki Mizutani Tokyo Medical and Dental University, Tokyo, Japan; RIKEN Research Center for Allergy and Immunology, Yokohama, Japan; National Research Institute for Child Health and Development, Tokyo, Japan [bold]Background: [/bold]Classical 3[beta]-hydroxysteroid dehydrogenase (3[beta]-HSD) deficiency is caused by loss of function mutations in the [italic]HSD3B2[/italic] gene encoding Type II isozyme of 3[beta]-HSD. The enzyme has a key role for steroid biosynthesis by converting the [Delta]5-steroids to the [Delta]4-steroids in adrenal glands and gonads, and the patients with 3[beta]-HSD deficiency show impaired synthesis of glucocorticoids and mineral corticoids in adrenal glands with [Delta]5-steroids excess. In spite of impaired [Delta]4-steroids synthesis in adrenal glands, it has been reported that in some 3[beta]-HSD deficiency patients, the serum level of 17-hydroxyprogesterone (17-OHP), one of the [Delta]4-steroids, is paradoxically elevated, but the precise mechanisms are not identified.[br][bold]Clinical case:[/bold] The patient (46, XX) was discovered by the newborn screening for 21-hydroxylase deficiency (21-OHD) with elevated serum level of 17-OHP (203 nmol/l). In spite of 17-OHP excess, the laboratory examinations showed remarkably increased [Delta]5-steroids [17-OH pregnenolone: 910 nmol/l, dehydroepiandrosterone (DHEA): 263 nmol/l], and other [Delta]4-steroids were not elevated, suggesting the patient has 3[beta]-HSD deficiency rather than 21-OHD.[br][bold]Mutational analysis: [/bold]Consistent with the endocrinological findings, we identified compound heterozygote for two novel missense mutations (p.Y190C and p.S218P) in [italic]HSD3B2[/italic], and each mutation were documented in both parents. Both amino acids were conserved among mammals, and, interestingly, located between two substrate binding sites, suggesting both mutations might affect the binding affinities for the substrate, [Delta]5-steroids. Consistently, a computational analysis of three-dimensional model structure of 3[beta]-HSD showed that both mutations were located in the predicted substrate-binding pocket. Thus, we hypothesized that the enzymatic activities of the two mutant proteins were diverse according to the substrates, [Delta]5-steroids. Indeed, the enzymatic activity of mutant protein for 17-OH pregnenolone were maintained significantly higher than that for other [Delta]5-steroids by [italic]in vitro[/italic] enzymatic activity assay, and we suggest it might cause the elevated serum levels of the metabolite from 17-OH pregnenolone, 17-OHP.[br][bold]Conclusion[/bold]: We identified two novel miss-sense mutations of the [italic]HSD3B2 [/italic]gene. In both mutants, the enzymatic activity for 17-OH pregnenolone were maintained significantly higher than that for other [Delta]5-steroids [italic]in vitro[/italic], suggesting the possible cause for elevated levels of 17-OHP in 3[beta]-HSD deficiency patients.[br][br]Nothing to Disclose: KT, KK, MO, YM, AH, NK, MT, OO, TM, SM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 408 71 703 SAT-622 PO54-01 Saturday 642 2012


643 ENDO12L_SAT-623 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Unusual Presentation of Congenital Adrenal Hyperplasia Due to 21-Hydroxylase Deficiency in a Newborn with Ambiguous Genitalia and Abdominal Mass Shruti Fadia, Jean-Pierre de Chadarevian, Francesco De Luca, Elizabeth Suarez St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA; St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA Background: 21-hydroxylase deficiency is the most common form of Congenital Adrenal Hyperplasia (CAH). Female infants with classic CAH due to 21-hydroxylase deficiency typically present with ambiguous genitalia. Bilateral diffuse enlargement of the adrenal glands may be detectable on imaging studies in patients with CAH. Here, we report the case of a newborn female with CAH presenting with a palpable mass detected on abdominal exam.[br]Clinical Case: B.T., born at 31 3/7 weeks, was the product of a twin gestation who presented with an abdominal mass and ambiguous genitalia at birth. Physical exam findings revealed a left lower quadrant abdominal mass, clitoromegaly, and nonpalpable gonads. Initial abdominal ultrasound demonstrated the presence of a uterus and a large multi-loculated cystic mass within the abdomen of unclear etiology. CT scan of the abdomen confirmed the presence of the mass, but the adrenal glands were not visualized. Further evaluation showed a normal female 46 XX karyotype, and high levels of testosterone (1630 ng/dL), 17-hydroxyprogesterone (45,900 ng/dL), and plasma renin activity (12,302 ng/dL/h) suggestive of CAH due to 21-hydroxylase deficiency. She was started on hydrocortisone and fludrocortisone therapy. On the fifth day of life, the patient underwent surgical excision of the left-sided retroperitoneal mass, which measured 11 cm x 10 cm x 3.5 cm. Pathology of the mass revealed the presence of giant, multicystic, hyperplastic, adrenocortical cells; there were no medullary cells or malignant cells identified. Abdominal ultrasound obtained one month post-operatively showed a grossly normal right adrenal gland, but no definite left adrenal gland. Abdominal MRI, which was obtained at age 2 years and 9 months, also failed to identify the left adrenal gland.[br]Conclusion: This is an unusual presentation of CAH in the newborn period. The significant left adrenal hyperplasia and absence of medullary cells may point towards a developmental abnormality of the adrenal gland in our patient.[br][br]Nothing to Disclose: SF, J-PdC, FDL, ES 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1554 71 704 SAT-623 PO54-01 Saturday 643 2012


644 ENDO12L_SAT-624 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Infant with WT1 Mutation, Ambiguous Genitalia and Congenital Diaphragmatic Hernia Michelle Y Rivera-Vega, Francis X Schneck, Suneeta Madan-Khetarpal, Miguel Reyes-Mugica, Selma Feldman Witchel Children[apos]s Hospital of Pittsburgh, Pittsburgh, PA [underline]Background:[/underline] Mutations in the WT1 gene are associated with Denys-Drash, Frasier, WAGR, and Meacham Syndromes. Denys-Drash Syndrome (DDS) is characterized by GU anomalies, progressive glomerulopathy, and Wilms tumor. Congenital diagragmatic hernia (CDH) have been reported in DDS with R366H mutation (1). FS is associated with progressive glomerulopathy, 46,XY karyotype, and WT1 intron 9 splice donor site mutations (2). We describe a full term 46,XX infant with ambiguous genitalia, CDH, and WT1 intron 9 splice acceptor site mutation.[br][underline]Clinical Case:[/underline] Infant, born to a 23 year old G2P1 female at 39+ weeks GA, had a prenatal diagnosis of left CDH. Amniocentesis had shown normal 46,XX karyotype, normal oligoarray, and negative prenatal FISH for trisomy 13, 18, or 21. She was intubated immediately after birth and transferred to the NICU. Exam showed an enlarged phallus, single perineal opening, ventral phallic grove, and fused non-rugated labioscrotal folds. Gonads were not palpated. Laboratory evaluation revealed: 46,XX karyotype, testosterone 12 ng/dl, ACTH stimulation test excluded diagnosis of congenital adrenal hyperplasia, and renal ultrasound showed bilateral renal cysts. Abdominal and pelvic MRI showed bicornuate uterus. Genotype analysis of WT1 showed a G to A transition at intron 9 (IVS9-1) which is predicted to generate a splicing mutation and a C to T transition at nucleotide 963 which is predicted to generate synonymous variant at codon 321. At 12 months of age, she had stable renal function, hypertension, minimal proteinuria, and no change in renal cysts.[br][underline]Conclusion:[/underline] Denys-Drash, Frasier, WAGR, and Meacham Syndromes are malformation syndromes associated with WT1 mutations. WT1 plays an important role in development of the urogenital system. The phenotype of WT1 knockout mice includes fetal lethality, aberrant urogenital development, and CDH (3). The mutation identified in our patient alters the intron 9 splice acceptor site. In a 46, XY patient with focal segmental glomerulosclerosis and bilateral cryptorchidism, this mutation activated an upstream splice acceptor site to generate a premature stop codon in exon 10 (4). Patients with 46,XX karyotype and WT1 mutations are uncommon and primarily manifest nephrotic syndrome and renal failure (5). How this specific mutation influences our patient[apos]s risk for Wilms tumor, gonadoblastoma, and renal disease is unknown. Our patient expands the phenotypic spectrum associated with WT1 mutations (6,7).[br][br]1. Antonius T et al. Amer J Med Genet, Part A,2008;146A:496. 2. Barbaux S et al., Nat Genetics 1997;17:467. 3. Kreidberg JA et al., Cell 1993;74:679. 4. Kanemoto K et al., Pediatr Nephrol 2007;22:454. 5. Fujita S et al., Clin Nephrol 2010:73:487. 6. Gwin K et al., Ped Develop Path 2008;11:122. 7. Chernin G et al. Clin J Am Soc Nephrol 2010;5:1655.[br][br]Nothing to Disclose: MYR-V, FXS, SM-K, MR-M, SFW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 695 71 705 SAT-624 PO54-01 Saturday 644 2012


645 ENDO12L_SAT-625 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Partial Androgen Insensitivity Syndrome (PAIS) with Subsequent Precocious Puberty in a Toddler Grace C Dougan, Dorothy I Shulman, Dorothy I Shulman University of South Florida, St Petersburg, FL BP was first examined at 6.5 mos for followup of PAIS previously diagnosed. At birth, hypospadias was present with urethral opening at the base of a 2.5 cm phallus. Scrotum was small but fused. Testes were palpable in the inguinal region. On day 2, FSH 1.3 mIU/mL, LH 11.3 mIU/mL, testosterone (T) 448 ng/dL, dihydrotestosterone (DHT) 111 ng/dL and karyotype was 46 XY. At 3 mos, estradiol (E) 26 pg/mL, FSH 3.1 mIU/mL, LH 16.9 mIU/mL, T 1300 ng/dL and DHT 405 ng/dL. Genetic testing of the androgen receptor revealed 2 mutations: a hemizygous c.1743 G-to-T change in exon 2, and a hemizygous c.1788 C-to-T change in exon 3 which was predicted to be deleterious by silico analysis. BP received T 25 mg injections at 3.5 and 4.5 mos. At 5 mos, penile length was 3 cm. He received an additional dose of T 50 mg at 5.5 mos at which time pubic hair was noted. Initial evaluation revealed length and weight at the NCHS 67th and 53rd percentiles, respectively. Testes were 0.5 cm in the inguinal canals. The scrotum was hypoplastic but hyperpigmented. Hypospadias and a chordee were noted; phallus was 3.3 cm x 1.7 cm. There was Tanner 3 pubic hair and no palpable breast tissue.[br]At 22 mos, L and W were at the NCHS [gt]99th percentile. Body hair was increased on his thighs and upper lip. L testes was 2 cm in inguinal canal and R testes was 2 cm x 3 cm, enlarged. Phallus was 4.5 cm x 2.5 cm after 2 repairs of hypospadias. Pubic hair remained Tanner stage 3. Breast tissue was absent. LH was 6.54 mIU/mL, FSH 0.32 mIU/mL, androstenedione (A) 34 ng/dL, 17-OHP 51 ng/dL, IGF-1 143 ng/mL and T 572 ng/dL. Bone age (BA) was 5 years. Pituitary MRI was nl. Testicular US confirmed bilateral enlargement with dimensions: R 2.6 cm x 1.5 cm x 1.4 cm and L 2.7 cm x 1.3 cm x 1.3 cm. Following leuprolide (20 mcg/kg) LH rose from 5 to 47 mIU/mL at 60 min. FSH rose from 3.1 to 5.4 mIU/mL; T 758 ng/dL, DHAS 59 ng/dL, inhibin B 286 pg/mL, SHBG 90 nmol/L, AMH 116 ng/mL, A 74 ng/dL, E 6.4 pg/mL. A dose of depot leuprolide (DL) was trialed given the BA advancement and elevated gonadotropins.[br]Here we report a pt with undervirilized genitalia and severe hypospadias. Genetic testing and laboratory evaluation were consistent with PAIS. The elevated T at 22 mos along with growth acceleration, BA advancement and persistence of pubic hair are unusual and unexplained. If there is little biochemical response to DL, further investigation for a concomitant gonadotropin independent process in the testes is planned.[br][br]Nothing to Disclose: GCD, DIS, DIS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2121 71 706 SAT-625 PO54-01 Saturday 645 2012


646 ENDO12L_SAT-626 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Management of 17[beta]HSD-3 Deficiency: Should Girls Remain Girls? Janet Chuang, Amy Vallerie, Lesley Breech, Crawford Peggy, Shumyle Alam, Pramod P Reddy, Howard Saal, Meilan M Rutter Cincinnati Children[apos]s Hospital Medical Center, Cincinnati, OH; Cincinnati Children[apos]s Hospital Medical Center, Cincinnati, OH; Cincinnati Children[apos]s Hospital Medical Center, Cincinnati, OH; Cincinnati Children[apos]s Hospital Medical Center, Cincinnati, OH; New York Medical College, Valhalla, NY; UC Physicians/University of Cincinnati, Cincinnati, OH Background: 17[beta]-Hydroxysteroid dehydrogenase type-3 (17[beta]HSD-3) deficiency is a rare cause of 46,XY disorder of sex development (DSD). The enzyme converts androstenedione (A) to testosterone (T), necessary for masculinization of male genitalia in utero. 17[beta]HSD-3 deficiency frequently presents late, following virilization at puberty, with resultant female-to-male gender reassignment in about 50%. The decision for sex of rearing is difficult, especially if diagnosed in early childhood. Consensus guidelines are equivocal or support male gender assignment. Long-term outcome data to guide decisions are lacking, and report mostly patients diagnosed after puberty. However, in the few cases of early diagnosis and orchiectomy, female gender retention appears more likely. The following 2 cases present consideration of female gender of rearing in conjunction with early gonadectomy in 17[beta]HSD-3 deficiency.[br]Case 1 presented at age 1y with female genitalia and bilateral inguinal herniae. Testes were found at surgery. Karyotype was 46,XY. She was initially diagnosed with complete androgen insensitivity syndrome (CAIS); however, androgen receptor mutation analysis was normal. HCG stimulation yielded a low T/A ratio (0.6, normal [gt]0.8). Genetic testing confirmed compound heterozygosity for 2 known mutations of [italic]HSD17B3[/italic]. Her parents chose to continue raising her as female. She underwent bilateral orchiectomy at age 2y.[br]Case 2 was born with female genitalia, virilized at age 13y, but did not seek evaluation until age 22y. Karyotype was 46,XY and she had bilateral inguinal testes. She had low T/A of 0.33. [italic]HSD17B3[/italic] analysis showed compound heterozygosity for 2 known mutations. She chose to maintain female gender, underwent bilateral orchiectomy, and started estrogen supplementation.[br]Clinical lessons and conclusions: Case 1 illustrates the difficulties faced with diagnosis and gender (re)assignment in 17[beta]HSD-3 deficiency at a young age. Both cases demonstrate that female gender may be preferred by the patient or family. When diagnosed early, orchiectomy may be appropriate to prevent virilization at puberty and establish female gender. These cases also highlight the importance of accurate diagnosis, given 17[beta]HSD-3 deficiency may have a female phenotype similar to CAIS, as this greatly influences gender reassignment decisions and counseling. Given the complexities involved, a multi-disciplinary DSD team is vital for establishing appropriate diagnosis, management and follow-up.[br][br]Nothing to Disclose: JC, AV, LB, CP, SA, PPR, HS, MMR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1737 71 707 SAT-626 PO54-01 Saturday 646 2012


647 ENDO12L_SAT-627 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Iatrogenic Cushing Syndrome in an Infant Secondary to Use of a Potent Topical Steroid for Diaper Rash Naziya Tahseen, Sripriya Raman Children[apos]s Mercy Hospitals and Clinics, UMKC School of Medicine, Kansas City, MO [bold]Background: [/bold]Exogenous steroid use is the most common cause of Cushing[apos]s syndrome. Topical steroids, although generally considered safer than systemic steroids, can cause Cushing[apos]s syndrome in vulnerable patients.[br][bold]Case:[/bold] We present a 3 month old previously healthy male who was referred for evaluation of cushingoid features. At six weeks of age he developed a diaper rash, following two days of watery diarrhea. Although his diarrhea resolved, the rash did not improve despite using over the counter creams including 1% hydrocortisone. Upon consultation with an adult dermatologist, he was prescribed Clobetasol Propionate 0.05% cream (class-I, around 600 times more potent than hydrocortisone). Parents were instructed to apply this to diaper area with occlusive dressings at each diaper change. Despite liberal use of Clobetasol cream (4 tubes, 15 gm each) for 6 weeks, the rash worsened. During this period he also developed increased appetite, rapid weight gain, disturbed sleep and frequent spitting episodes. He was seen by a pediatric dermatologist who discontinued Clobetasol cream and referred him to the pediatric endocrinology urgent clinic for further management. Physical exam was significant for moon facies, flushed cheeks, severe diaper dermatitis with skin excoriation, and no linear growth over past 4 weeks. He was diagnosed with iatrogenic Cushing[apos]s syndrome and presumed HPA axis suppression after acute withdrawal of chronic exogenous steroid. We treated with oral hydrocortisone (30mg/m2) that was gradually weaned over 6 weeks. After nephrology consultation, we used chlorothiazide for persistent hypertension. At follow-up visit cushingoid features and diaper rash were significantly improved. Blood pressure returned to normal range after 2 weeks. To evaluate recovery of his HPA axis, an ACTH stimulation test will be done 6 weeks after discontinuation of hydrocortisone.[br][bold]Conclusions:[/bold] Clobetasol cream is contraindicated in infants and children due to serious systemic adverse effects such as Cushing[apos]s syndrome and concomitant HPA axis suppression. However, Clobetasol continues to be used inappropriately, and in some countries it is easily available for purchase without a prescription. Physicians should be aware of the safe and limited use of topical steroids; they should also educate patients and parents of the potential side effects.[br][br]Nothing to Disclose: NT, SR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 675 71 708 SAT-627 PO54-01 Saturday 647 2012


648 ENDO12L_SAT-628 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) A Pediatric Cushing Patient with Ectopic ACTH Syndrome and Concomitant Pituitary Microadenomas Alyse S Goldberg, Robert Stein, Neil Merrit, Richard Inculet, Stan Van Uum Schulich School of Medicine and Dentistry, University of Western Ontario, London, Canada; Schulich School of Medicine and Dentistry, University of Western Ontario, London, Canada; Schulich School of Medicine and Dentistry, University of Western Ontario, London, Canada; Schulich School of Medicine and Dentistry, University of Western Ontario, London, Canada Introduction: Ectopic ACTH syndrome (EAS) is a rare but important cause of pediatric Cushing[apos]s syndrome (CS). Although diagnostic evaluation of CS is similar to that in adults, the variation in epidemiology (pituitary ACTH producing adenoma being very common and EAS being rare) may sway investigations leading to inappropriate diagnostic interventions.[br]Case report: We present a 15-year-old girl presenting with classic signs and symptoms CS. Diagnostic investigations confirmed CS by urinary free cortisol 9855 nmol/day (N[lt]346) and an elevated midnight salivary cortisol (127 nmol/L; N[lt]5nmol/L). Severe hypokalemia (2.7 mmol/L) and a random blood sugar of 23.6 nmol/L illustrated the severity. Elevated serum ACTH concentrations (42.4 pmol/L at 8 am; N[lt]12.47) confirmed an ACTH dependent form of CS and prompted investigations to locate the ACTH source. A MRI of the sella identified two pituitary adenomas (4 and 2mm). Petrosal venous sampling aimed at lateralizing ACTH production, identified an ectopic source. Subsequently, an octreotide scan demonstrated increased uptake in the left lower lobe of the lung and in a para-esophageal lymph node, suspicious for carcinoid tumor. Therapeutic management of this stage 3A thoracic carcinoid tumor consisted of radical surgical excision of the primary tumor and mediastinal node dissection. Close perioperative monitoring of blood pressure, potassium, glycaemia and cortisol status was crucial. Hypertension and hyperglycemia resolved during hospitalization, while weaning off supplemental glucocorticoids took 6 months. Success of surgical resection was assessed by biochemical resolution and negative Indium 111 Octreotide scan post-operatively[br]Conclusion: Although rare, EAS is an important cause of pediatric CS and thus must be considered in diagnostic workup when clinical suspicion is present. Even in the pediatric population, a pituitary lesion [lt]6 mm on MRI should not be considered sufficient confirmation of Cushing[apos]s disease, and appropriate diagnostic work up should be performed. Perioperative management of patient with EAS is challenging since potassium, glycaemia and cortisol status are tenuous once the ACTH source is removed.[br][br]Nothing to Disclose: ASG, RS, NM, RI, SVU 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 85 71 709 SAT-628 PO54-01 Saturday 648 2012


649 ENDO12L_SAT-629 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Pseudohypoaldosteronism: A Rare Cause of Salt Wasting in the Newborn Angela Santiago-Lennon, Gunjeet Kala Ahluwalia University of Kansas Medical Center, Kansas City, KS Introduction: Pseudohypoaldosteronism (PHA) Type I is a rare disease characterized by mineralocorticoid unresponsiveness. The main treatment involves large amounts of sodium and fluid replacement, which help reverse accompanying electrolyte abnormalities. Genetic testing is available to locate the defect in the mineralocorticoid receptor or in the epithelial sodium channel (ENaC), which differ in clinical course. We present a case of an infant with multisystemic PHA Type I, with 2 novel mutations in the [alpha] subunit of the ENaC.[br]Case report: A 1 month old Caucasian male, born at 36 weeks to non consanguineous parents, presented at 1 wk of life with skin mottling, emesis and lethargy. Lab work up showed hyponatremia (Na 123mEq/L), hyperkalemia (K 9meq/L), and metabolic acidosis (HCO[sub]3[/sub] 13meq/L). Urine electrolytes showed high Na and low K excretion. 12L EKG done was normal. The infant was managed with fluid resuscitation, sodium replacement (NS and oral NaCl), kayexylate, insulin drip and dextrose, NaHCO3, hydrocortisone and fludrocortisone. The diagnosis of pseudohypoaldosteronism was eventually made after initial labs showed elevated renin activity at 93.96 ng/ml/hr (range 2-37), and aldosterone 587.6 ng/dl (normal [lt]217). Normal cortisol (12.7 [micro]g/dl[italic]) [/italic]and 17-OH progesterone (20 ng/dl) ruled out defects in steroidogenesis. To rule out secondary causes of PHA, renal ultrasound was done, which revealed slightly smaller kidneys. VCUG was negative. The infant was soon noted to have tachypnea and mild rales, and required oxygen. Chest xray showed mild pulmonary congestion. He was started on hydrocholorothiazide and albuterol, to which he responded well. Systemic type of PHA was then suspected, and a sweat test obtained came back elevated (118mmol/L). CF gene testing was negative. The infant was discharged at 2 mos. of age on an oral regimen of NaCl, Kayexalate and Na citrate (total Na 5.4 g/day). Genetic testing subsequently showed 2 novel mutations in the SCNN1A gene ([alpha]-ENaC), c.416G[gt]A (p.Arg139Lys) exon 2 and c.1360+1G[gt]T(exon 8 splice donor site). Patient is currently 7 mos. of age and continues to do well, despite a brief readmission for hyperkalemia and a mild episode of URI which was treated with albuterol at home.[br]Discussion: Multisystemic PHA Type I results from autosomal recessive mutations in the [alpha], [beta], or [gamma] subunits of the ENaC. As a result of the widespread location of this channel, organs like the lungs, sweat glands and colon may also be involved.[br][br]Nothing to Disclose: AS-L, GKA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 267 71 710 SAT-629 PO54-01 Saturday 649 2012


650 ENDO12L_SAT-630 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Isolated Aldosterone Deficiency in a Non-Consanguineous Family of Southern Indian Origin Christopher John Dunne, Shruti Fadia, Rita Ann Kubicky, Francesco De Luca, Elizabeth A Suarez St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA [bold]Background:[/bold] Isolated Aldosterone Deficiency is a rare cause of failure to thrive that has been described in Iranian Jewish, Pakistani, and Caucasian populations. We describe a case of hyper-reninemic hypo-aldosteronism in a patient who was the product of non-consanguineous parents of Southern Indian ethnicity.[br][bold]Clinical case:[/bold] A 28-day old male was admitted at St. Christopher[apos]s Hospital for Children for failure to thrive. He was born full-term, with normal birth weight and length (3.26 kg and 53.3 cm respectively). At his newborn checkup, he weighed less than his birth weight despite seemingly adequate caloric intake; thus, he was admitted for further workup.[br]On admission, his weight was 3.1kg (3[sup]rd[/sup] percentile), length 55cm (25[sup]th[/sup] percentile), and his head circumference 35cm (5[sup]th [/sup]percentile). On exam, he had no dysmorphic features or midline defects and normal male genitalia with testes descended bilaterally. Laboratory studies revealed significantly low sodium of 115 mmol/L (nl. 134-144), high potassium of 6.8 mmol/L (nl. 3.4-6.0), normal anion gap of 9, BUN of 27 mg/dL (nl. 4-12) and creatinine of 0.42 mg/dL (nl. 0.3-1.0). The patient received an initial IV sodium correction, and over the next 48 hours he continued to require IV normal saline solution in order to maintain normal sodium levels.[br]On hospital day 3, he was found to have undetectable plasma aldosterone and high plasma renin activity of 65.7 ng/mL/hr (nl. 2.0-37.0). He was started on sodium chloride (1 gram po/day) and fludrocortisone (0.1 mg bid) supplementation. Over the next few days his sodium and potassium normalized (136 mmol/L and 5.1 mmol/L), and he was weaned off IV NSS supplementation.[br]A Cosyntropin stimulation test showed high stimulated levels of deoxycorticosterone of 347 ng/dL (nl. 40-158), elevated corticosterone of 11,704 ng/dL (nl. 2225-4974), and normal 18- hydroxycorticosterone of 359 ng/dL (130-465) consistent with the diagnosis of Aldosterone Synthase Deficiency. Stimulated levels of cortisol, 17-OH progesterone, and 17-OH pregnenolone were normal excluding other adrenal causes of salt-wasting.[br][bold]Conclusion:[/bold] To our knowledge, this is the first case of isolated Aldosterone deficiency described in a patient of Southern Indian ethnicity. Aldosterone deficiency remains a rare, yet possible, cause of hyponatremic failure to thrive, even in non-consanguineous families. Genetic studies to define the etiology of this patient[apos]s Aldosterone deficiency are warranted.[br][br]Nothing to Disclose: CJD, SF, RAK, FDL, EAS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1928 71 711 SAT-630 PO54-01 Saturday 650 2012


651 ENDO12L_SAT-631 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Growth Hormone Treatment for Short Stature in a Patient with Pseudohypoaldosteronism Type II Emir Tas, Claudia C Boucher-Berry University of Illinois Medical Center at Chicago, Chicago, IL; University of Illinois Medical Center at Chicago, Chicago, IL Background: Pseudohypoaldosteronism type II (PHAII) is a unique autosomal dominant inherited cause of familial hypertension associated with hyperkalemia. Normal glomerular filtration rate with suppressed renin activity and relatively low aldosterone concentrations are common features of the disease. Short stature, although not a component of the syndrome, has been described in a few sporadic case reports. There are no reported cases of children with PHAII being treated with growth hormone.[br]Case Report: MF is an 8 year old Israeli female who was diagnosed with PHAII shortly after birth. Due to a history of known PHAII on the maternal side of the family, MF underwent genetic testing shortly after birth. She was found to have a missense mutation (Gln565Glu) detected on the Serine/threonine-protein kinase WNK4 gene on the long arm of chromosome 17. This mutation results in a change in the predicted amino acid sequence from neutral and polar glutamine (Gln) to the negatively charged glutamic acid (Glu).[br]Her growth for the first three years of life remained below the third percentile with a growth velocity of 4 cm/year. At three years of age, work-up for short stature was negative for Celiac Disease and Cystic Fibrosis. Growth hormone stimulation testing performed using Clonidine showed a peak growth hormone level of 21 ng/ml. Her growth velocity was continually monitored until 6 years of age, when she presented with hypertension and hyperkalemia. Although she was started on Hydrochlorothiazide, with good control of the hypertension and hyperkalemia, her growth velocity continued to be low. She was started on growth hormone therapy at a dose of 0.035 mg/kg/week and in the following year, the growth velocity has increased to 9 cm/year. She has not required any increase in the Hydrochlorothiazide dose and is not having any adverse effects from the growth hormone.[br]Discussion: An important and rare feature of PHAII mentioned mainly in sporadic case reports is short stature. We have described an 8 year old prepubertal girl with short stature diagnosed with PHAII at birth. Patients in previous case reports were managed with dietary salt restriction and thiazide diuretics and were reported to have catch-up growth. Growth hormone therapy has never been tried in this patient population to our knowledge. This report extends the repertoire of conditions that respond to growth hormone and suggests that GH may be safely used in treatment of short stature in patients with PHAII.[br][br]Nothing to Disclose: ET, CCB-B 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1300 71 712 SAT-631 PO54-01 Saturday 651 2012


652 ENDO12L_SAT-632 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Pseudohypoaldosteronism Type II and Growth Hormone Deficiency in a School Age Child Presenting with Hyperkalemia and Short Stature Kelly A Hicks, Ioanna Athanassaki, Sarah Swartz Texas Children[apos]s Hospital and Baylor College of Medicine, Houston, TX; Texas Children[apos]s Hospital and Baylor College of Medicine, Houston, TX Background: Children with pseudohypoaldosteronism type II (familial hyperkalemic hypertension or Gordon[apos]s syndrome) exhibit hyperkalemia, metabolic acidosis, and short stature. Pre-hypertension and normotension have been noted in the pediatric age group.[br]Clinical Case: An 11 year old female presented to a pediatric emergency room for evaluation of hyperkalemia associated with palpitations, dizziness, and fatigue. During a prior well child exam notable for poor growth, she was found to be hyperkalemic with a serum potassium of 7.6 mmol/L (3.8-5.1 mmol/L), sodium 138 mmol/L (135-146 mmol/L), and a slight metabolic acidosis with chloride 114 mmol/L (98-110 mmol/L) and bicarbonate 16 mmol/L (21-33 mmol/L). Repeated electrolytes upon arrival to the emergency room confirmed potassium 6.6 mmol/L but no evidence of metabolic acidosis (bicarbonate 21 mmol/L). Electrocardiography displayed peaked T waves. She was normotensive for age and height with blood pressures 110/59 and 100/62. Renin was suppressed at 0.3 ng/ml/hr (0.5- 3.3 ng/mL/hr) and serum aldosterone was normal at 16 ng/dl ([lt] OR = 21 ng/dl) with a corresponding serum sodium of 135 mmol/L. Additional studies indicated a low trans-tubular potassium gradient (TTKG) at 1.75 and an elevated aldosterone to renin ratio (ARR) at 53. A 1 mcg ACTH stimulation test was normal with a peak cortisol level of 27.1 [micro]g/dl at 60 minutes. Pseudohypoaldosteronism type II was diagnosed based on the constellation of poor growth, hyperkalemia, low TTKG, and elevated ARR. No other affected relatives were identified. Treatment with a thiazide was initiated at 20 mg/kg/day with adjustments based on subsequent serum electrolytes. Height and weight were both below the 5% for age, and bone age was delayed with skeletal age estimated at 8 years by the standards of Greulich and Pyle. Growth velocity was unchanged in the six months following diagnosis despite the patient[apos]s peri-pubertal age (Tanner II). A growth hormone stimulation test to clonidine and glucagon was performed with maximal response to clonidine 5.9 ng/ml at 60 minutes and maximal response to glucagon 0.7 ng/ml at 180 minutes. Growth hormone therapy was initiated at 0.3 mg/kg/week.[br]Conclusion: A thiazide diuretic successfully managed hyperkalemia in a patient with pseudohypoaldosteronism type II. Growth hormone deficiency has been reported in association with autosomal recessive pseudohypoaldosteronism type I but not in association with pseudohypoaldosteronism type II.[br][br]Nothing to Disclose: KAH, IA, SS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 200 71 713 SAT-632 PO54-01 Saturday 652 2012


653 ENDO12L_SAT-633 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Congenital Adrenal Hypoplasia Initially Presenting with Aldosterone Deficiency in a Child Conceived with a Donated Egg: A Case Report and Ethical Dilemma Heba Ismail, Marielisa Rincon Seattle Children[apos]s Hospital, Seattle, WA; TC Thompson Children[apos]s Hospital, Chattanooga, TN [bold]Background: [/bold]X-linked [italic]Congenital Adrenal Hypoplasia[/italic] is a rare adrenal cortical disorder caused by NROB1/DAX-1 gene mutation. It classically presents with simultaneous glucocorticoid and mineralocorticoid deficiency. This is the first case report of such condition in a child conceived with a donated egg.[br][bold]Clinical Case: [/bold]A 34 weeks male newborn, fraternal twin, conceived by invitro fertilization of donated eggs, presented shortly after birth with feeding intolerance and electrolyte abnormalities (hyponatremia and hyperkalemia). Newborn screening results demonstrated a normal 17-hydroxyprogesterone. Further testing revealed a low serum aldosterone, high plasma renin activity, along with normal serum cortisol levels. He was diagnosed with 18 hydroxylase deficiency based on low 18 hydroxycorticosterone levels and was therefore treated with mineralocorticoid replacement. He had an excellent response to therapy and remained well controlled for 18 months.[br]At 18 months of age, the patient presented to the ED with dehydration secondary to herpetic gingivostomatitis. At the time of presentation, he was also found to have hypoglycemia, hyponatremia, hyperkalemia, acidosis and a low serum cortisol level (2.7 ug/dl). A low dose ACTH stimulation test revealed no post-stimulation cortisol changes. His ACTH level at the time was extremely elevated, with low levels of all adrenal cortex products. The patient was started on glucocorticoids successfully (hydrocortisone). Because of his course and biochemical profile, genetic testing for NROB1 mutation was pursued, confirming the diagnosis of [italic]Congenital Adrenal Hypoplasia.[/italic][br]His asymptomatic fraternal twin was also tested for NROB1 mutation and results were negative. The fertility center was contacted and it was found in their records that the donor mother had two sisters but no brothers. She had donated eggs to other families with a mixture of male and female children, but none were known to have this disorder in particular. The egg donor was informed by the fertility center, but moved out of state and to our knowledge did not pursue genetic testing.[br][bold]Conclusions: [/bold]We report a case of [italic]Congenital Adrenal Hypoplasia[/italic] presenting in the context of extraordinary ethical considerations. This case raises a question that is unique to this modern era of assisted reproduction: should we screen for rare but yet potentially life threatening genetic conditions in both egg and sperm donors prior to pursuing donation?[br][br]Nothing to Disclose: HI, MR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 89 71 714 SAT-633 PO54-01 Saturday 653 2012


654 ENDO12L_SAT-634 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Germ Cell Tumors and 46,XY DSD Kanthi Bangalore-Krishna, Melissa Buryk, Selma F Witchel Children[apos]s Hospital of Pittsburgh of UPMC/University of Pittsburgh, Pittsburgh, PA [bold][underline]BACKGROUND[/underline][/bold]: Patients with partial 46,XY DSD are often identified during infancy due to atypical development of external genitalia; these patients have a high risk for neoplastic changes of the dysgenetic gonad(s). Patients with 46, XY complete gonadal dysgenesis (CGD) or Swyer syndrome may have normal female external genitalia, intra-abdominal streak gonads and presence of Mullerian structures. These patients may not present until adolescence when they are referred for the evaluation of delayed puberty and primary amenorrhea. We report 2 patients who presented with germ cell tumors; subsequent karyotype analysis showed 46,XY.[br][bold][underline]CASE 1[/underline][/bold]: A 12 year old Caucasian female initially presented to the emergency room with abdominal pain. Abdominal ultrasound and subsequent laparoscopy revealed a right ovarian dysgerminoma, absent left gonadal tissue, and a uterus. Following surgery, a karyotype showed 46,XY (positive for SRY); she was then referred to endocrinology for evaluation. She was tall with Tanner II breasts, Tanner III pubic hair, and normal female external genitalia. Laboratory evaluation revealed FSH 133.5 mIU/ml, LH 52.2 mIU/ml, and ultrasensitive estradiol [lt]5 pg/ml consistent with gonadal failure.[br][bold][underline]CASE 2:[/underline][/bold] A 15 year old Caucasian female was referred to endocrinology for the evaluation of delayed puberty and primary amenorrhea. At 11 years of age, she presented with abdominal pain due to a left ovarian germ cell tumor (germinoma and immature teratoma) which was treated with chemotherapy and resection. On exam, she was tall with Tanner III breasts, Tanner II pubic hair and normal female external genitalia. Laboratory evaluation showed FSH 70.4 mIU/ml, LH 27.8 mIU/ml, and ultrasensitive estradiol [lt]2 pg/ml consistent with gonadal failure. Karyotype was 46XY (positive for SRY). Subsequent laparoscopy showed a right diminutive fallopian tube, streak gonad, a small uterus and cervix. Due to a high risk for malignant degeneration, the gonad was removed.[br][bold][underline]DISCUSSION:[/underline][/bold] Both patients had normal female external genitalia. Although, both presented with gonadal tumors, patient 1 was diagnosed soon after surgery whereas, for patient 2, the diagnosis was delayed for several years. Since approximately 30% of patients with 46,XY CGD develop gonadal tumors, karyotype analyses should be obtained among these individuals because of the risk for malignant degeneration (1). If the risk for malignancy is high, gonadectomy needs to be considered at the time of diagnosis (2,3).[br][br](1)Rocha VB, et al., Fertil Steril 2011;96:1431. (2)Doherty LF et al., Fertil Steril 2011;96:1415. (3)Zielinska D, et al., J Pediatr Surg 2007;42:1721.[br][br]Nothing to Disclose: KB-K, MB, SFW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 385 71 715 SAT-634 PO54-01 Saturday 654 2012


655 ENDO12L_SAT-635 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Autoimmune Hepatitis as the initial Presentation of Autoimmune Polyglandular Syndrome Type 1 in a Female Adolescent Lina Huerta-Saenz, Kavitha Dileepan, M Max Feldt, Alexander Karmazin The Children[apos]s Mercy Hospital and Clinics, Kansas City, MO [bold]Background:[/bold] Autoimmune polyglandular syndrome (APS) type 1 is an uncommon autosomal recessive disease characterized by mucocutaneous candidiasis, autoimmune hypoparathyroidism and adrenal insufficiency. Less commonly associated conditions are type 1 diabetes mellitus, vitiligo, alopecia, fat malabsorption, Ig-A deficiency, autoimmune hepatitis, red cell aplasia and progressive myopathy. APS type 1 occurs from mutations in the autoimmune regulator (AIRE) gene. Typically, initial presentation occurs during infancy with mucocutaneous candidiasis with other conditions developing later, usually within 20 years [sub](1).[/sub][br][bold]Clinical case:[/bold] A previously healthy 13-year old Caucasian female with negative family history was admitted with a 2 day history of progressive jaundice and abdominal pain. Autoimmune hepatitis type 2 with positive anti-LKM (anti-liver-kidney microsome) antibodies was diagnosed. She began treatment with glucocorticoids and immunosuppressive therapy. During her 35 day-hospitalization, she developed acute liver failure, severe hyperglycemia requiring insulin therapy and symptomatic hypocalcemia (numbness of the extremities). She was also found to have oroesophageal candidiasis. Endocrine evaluation revealed positive GAD and ICA-512/IA-2 antibodies (37.5 and 2.1 unit/mL respectively). An inappropriately normal PTH level was detected (28 pg/mL) at a serum calcium level of 7.5mg/dL. Hypoparathyroidism was diagnosed and her serum calcium rapidly improved on calcitriol. Auto-antibodies (thyroid, ovarian, adrenal, celiac) and AIRE gene mutation analyses were collected and were negative. During her hospital admission, hepatic function improved, glycemia remained under good control with insulin therapy, and eucalcemia was maintained with calcitriol. Interestingly, her AIRE gene analysis was ultimately negative. APS is a clinical diagnosis and genetic testing is not necessary to confirm diagnosis.[br][bold]Conclusion:[/bold] Autoimmune hepatitis is an uncommon initial presentation of APS type 1, and the detection of two additional autoimmune endocrinopathies within 35 days of presentation has not been previously reported. In this clinical case, clinical suspicion and screening for APS type 1 facilitated prompt diagnosis and successful clinical management. Further research into other genetic markers may help with the understanding of different clinical manifestations of APS.[br][br](1) Owen, C., Cheetham, T. Diagnosis and management of polyendocrinopathy syndromes. Endocrinol Metab Clin N Am 2009; 38; 419-436.[br][br]Nothing to Disclose: LH-S, KD, MMF, AK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1035 71 716 SAT-635 PO54-01 Saturday 655 2012


656 ENDO12L_SAT-636 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Delayed Diagnosis of Severe Hypertriglyceridemia in Childhood Jessica S Lilley, MacRae F Linton, Sergio Fazio Vanderbilt Children[apos]s Hospital, Nashville, TN; Vanderbilt University School of Medicine, Nashville, TN Background. Lipoprotein lipase deficiency causes a severe hypertriglyceridemia that typically presents in childhood with abdominal pain, eruptive xanthomas, lipemia retinalis, or neurologic symptoms such as irritability or learning difficulties. While xanthomas are not always present, recognizing them on physical exam can lead to timely diagnosis of devastating diseases.[br]Clinical case. A 7 year-old girl gave subtle clues throughout her life to an underlying disease prior to acute decompensation with pancreatitis and shock that led to diagnosis of severe hypertriglyceridemia. There was no family history of dyslipidemia, and she was taking no medications. She first presented to her pediatrician with yellow, waxy plaques on her face and extremities as an infant and was diagnosed with neonatal acne. The skin lesions persisted and worsened with time. She had progressive attention deficit which became evident once she entered school. She suffered from frequent epigastric abdominal pain for many years and consulted her pediatrician and eventually a pediatric gastroenterologist. She was treated for constipation with no relief. No labs were drawn at any of these visits.[br]In 2011, she presented to our hospital[apos]s ED in hypovolemic shock and was admitted to the pediatric intensive care unit. Initial lab samples were grossly lipemic, so a lipid panel was sent. After more than 24 hours of fasting, her triglycerides were 2549 mg/dL. Continued fasting with parenteral nutrition brought the triglycerides down to 150 mg/dL after 72 hours. Following discharge, she has maintained acceptable triglyceride levels with strict dietary modifications. Her mother notes that her facial xanthelasma recedes when her triglycerides are within the goal range.[br]Conclusion. Though pediatricians rarely encounter complications of dyslipidemias, the most severe cases often present in childhood. Training to identify skin manifestations of hyperlipidemia could have prevented a life-threatening admission for pancreatitis in this case.[br][br]Sources of Research Support: Dr. Lilley is funded by NIH grant 2T32DK007061-37.[br][br]Disclosures: MFL: Consultant, Pfizer, Inc., Merck & Co., Genzyme Corporation, Kowa. SF: Consultant, Pfizer, Inc., Kowa, Roche Diagnostics, Takeda, Merck & Co. Nothing to Disclose: JSL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1693 71 717 SAT-636 PO54-01 Saturday 656 2012


657 ENDO12L_SAT-637 POSTER SESSION: Pediatric Endocrine Case Reports: Diabetes, Hypoglycemia, Adrenal, Disorders of Sexual Development (1:30 PM-3:30 PM) Incidental Abdominal Mass in an Adolescent Girl: Multiple Endocrine Neoplasia Type 1 Naziya Tahseen, Julia R Broussard, Figen Ugrasbul Children[apos]s Mercy Hospitals and Clinics, UMKC School of Medicine, Kansas City, MO Background: Multiple Endocrine Neoplasia type 1 (MEN1) has a prevalence of 1-10/100,000. It has an autosomal dominant or sporadic inheritance. Hyperparathyroidism is the most common endocrinopathy affecting nearly 100% of patients; it is also the 1st presentation in 90% of cases. Enteropancreatic cell tumors (60-80%) and pituitary tumors (15-50%) are other important lesions in MEN1.[br]A 16 year old girl was evaluated for weight gain, irregular periods and occasional tachycardia for the duration of one year. Past medical history: Hypertension, headaches and chronic abdominal pain. Family history: Maternal grandaunt had pheochromocytoma. Several family members had parathyroidectomies. Physical examination: BMI was 37.5 with acanthosis nigricans of neck and axilla. Oral glucose tolerance test showed insulin resistance. Work-up for Cushing[apos]s disease and polycystic ovarian syndrome was negative. At follow up she had urinary tract infections and right lower abdominal pain. Abdominal ultrasound (US): left sided nephrolithiasis and an incidental mass in left upper quadrant. Magnetic Resonance Imaging (MRI) abdomen revealed a 9.1 x 8.9 x 9.0 cm heterogeneous mass arising from the body and tail of pancreas, representing a primary pancreatic neoplasm. Pheochromocytoma work up done for hypertension, tachycardia and abdominal mass was negative. Subsequently, she had surgical resection of the mass. Histology showed well differentiated insulinoma. This prompted a work up for MEN1. Labs: Calcium (Ca): 12.0 mg/dl (8.6 - 10.5), Ca Ionized: 1.42 mmol/L(1.13 - 1.37), PTH Intact: 98 pg/ml(7 [ndash] 75), Phosphorus: 3.5 mg/dl(2.3 - 4.8), Urine Ca/Cr ratio: 0.19([lt]0.2), Prolactin: 51.7 ng/ml(0.0 - 17.0), Serum Gastrin: 265 pg/ml([lt]100). Sestamibi scan of parathyroid was normal. Thyroid US: Hypoechoic nodule in left lobe of thyroid, suggestive of parathyroid adenoma. MRI Brain: Heterogeneously enlarged pituitary, consistent with pituitary adenoma. She had 4 gland parathyroidectomy and histology showed parathyroid hyperplasia. Cytogenetic analysis confirmed heterozygous deletion of the MEN1 gene.[br][underline]Conclusion[/underline]: Insulinoma arises in 10% of MEN1 patients, and usually presents in the third decade of life, as single or multiple macroadenomas 1-4 cm in diameter; 40% patients can be asymptomatic. Interestingly, our patient presented with a large asymptomatic insulinoma in her second decade of life. MEN1 can have variable presentations and diagnosis of sporadic cases requires high clinical suspicion.[br][br]Nothing to Disclose: NT, JRB, FU 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 779 71 718 SAT-637 PO54-01 Saturday 657 2012


658 ENDO12L_SAT-641 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Dominant-Negative Effect of PRR Insertion in the Intracellular Domain of KISS1R Associated with Isolated Hypogonadotropic Hypogonadism Lucie Chevrier, Eva Hernandez, Anne-Marie Guedj, Alexandre de Brevern, Nicolas de Roux Universit[eacute] Paris Diderot Paris 7, Paris, France; H[ocirc]pital Car[eacute]meau, N[icirc]mes, France; Universit[eacute] Paris Diderot Paris 7, Paris, France; H[ocirc]pital Robert Debr[eacute], Paris, France KISS1R and its ligand Kisspeptin were identified as a major regulator of the gonadotropic axis. Mutations of the KISS1R gene result in isolated hypogonadotropic hypogonadism (IHH).[br]We identified a novel heterozygous variant of KISS1R corresponding to the addition of one proline and two arginines in the intracellular domain of the receptor in a young boy suffering from IHH (p.A342_P343insPRR). This insertion is located in a proline/arginine-rich region leading to four consecutive PRR motifs instead of three. Functional analyses of this mutated receptor (PRR-KISS1R) in transiently transfected HeLa cell line indicated that the PRR insertion did not prevent intracellular signalization (PLC and MAPK pathways) but altered the intracellular trafficking of KISS1R by reducing cell surface expression without modifying total expression. Co-transfection of wild type (WT) and mutated receptors in HeLa cells demonstrated that PRR-KISS1R exerted a dominant-negative effect on the expression of WT-KISS1R. In-silico molecular dynamic study indicated that PRR insertion strongly altered the flexibility of proline and arginine-rich region and gave rise to spatial constraints, leading to a more rigid structure compared to WT-KISS1R. An alanine scan of this region combined with a sequence alignment analysis revealed that the highly conserved Arg344 is crucial for KISS1R normal expression in HeLa cells.[br]This study provides the first description of a KISS1R heterozygous mutation leading to IHH by a dominant-negative effect on the WT receptor. The proline/arginine-rich region located in the intracellular domain of KISS1R appears to play a key role in synthesis, intracellular traffic and activation of KISS1R. A more precise structure-function analysis of this region and the identification of partner proteins able to regulate KISS1R traffic or activation could lead to a better understanding of kisspeptins-KISS1R system activation at the onset of puberty.[br][br]Nothing to Disclose: LC, EH, A-MG, AdB, NdR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1438 72 719 SAT-641 PO17-01 Saturday 658 2012


659 ENDO12L_SAT-642 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Dynamic KISS1R Trafficking Modulates Kisspeptin Signaling Activity Le Min, Kathleen Soltis, Ana Claudia Reis, Shuyun Xu, Wendy Kuohong, Rona S Carroll, Ursula Kaiser Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA; University of S[atilde]o Paulo, S[atilde]o Paulo, Brazil Kisspeptin receptor (KISS1R) signaling plays a critical role in the central regulation of reproduction. We investigated the role of kisspeptin-stimulated KISS1R internalization, recycling, and degradation in the modulation of KISS1R signaling for wild-type (WT) KISS1R and for two mutant receptors identified in patients with central precocious puberty or hypogonadotropic hypogonadism. In HEK cells stably expressing WT or R386P KISS1R, a gain-of-function mutant, stimulation with kisspeptin-10 (kp10) resulted in a biphasic increase in intracellular calcium, with a rapid acute increase followed by a more sustained second phase. The R386P KISS1R demonstrated a more robust calcium response in both the first and second phases compared to WT KISS1R. In contrast, L148S KISS1R, a loss-of-function mutant, failed to respond. The second phase calcium response for both WT and R386P KISS1R was abolished by removal of kp10 from culture medium after stimulation. Notably, the second phase calcium response was also inhibited by dynasore, a dynamin inhibitor, suggesting that KISS1R internalization contributes to the second phase calcium response. To determine the effects of ligand on KISS1R trafficking, we measured kp10-stimulated effects on cell surface KISS1R using an ELISA. The results showed a decrease in cell surface receptors for WT and R386P KISS1R within 20 minutes after kp10 stimulation, which returned to baseline levels 20 minutes after removal of kp10. No changes were observed for L148S KISS1R. Finally, we investigated the fate of the internalized kisspeptin-KISS1R complex. Most internalized kisspeptin was released extracellularly in degraded form within one hour, suggesting rapid processing of the internalized kisspeptin-KISS1R complex. Using a biotinylation assay, we demonstrated that KISS1R degradation was much slower than the degradation of internalized kisspeptin, suggesting dissociated processing of the internalized kisspeptin-KISS1R complex that may contribute to the sustained exogenous ligand-dependent KISS1R calcium signaling. Taken together, our results suggest that the second phase calcium response to kp10 is dependent on continued presence of ligand and is the result of dynamic KISS1R trafficking. The diminished signaling and trafficking by L148S KISS1R further supports the coupling of KISS1R trafficking and signaling. The more robust and sustained calcium signaling response to kp10 by R386P KISS1R may contribute to the precocious puberty phenotype.[br][br]Sources of Research Support: NICHD.[br][br]Nothing to Disclose: LM, KS, ACR, SX, WK, RSC, UK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1621 72 720 SAT-642 PO17-01 Saturday 659 2012


660 ENDO12L_SAT-643 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) GnRH Pulse Frequency-Dependent Regulation of ICER, a Modulator of FSH[beta] Transcription, Is Attenuated by MEKI/II Blockade Iain R Thompson, Nick A Ciccone, Shuyun Xu, Qiongjie Zhou, Sofiya Zaytseva, Rona S Carroll, Ursula B Kaiser Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA The pulsatile release of the hypothalamic decapeptide GnRH activates signal transduction cascades in the pituitary gonadotrope to control the production and secretion of FSH and LH. The gonadotropins have distinct FSH[beta] and LH[beta] subunits, which are preferentially stimulated at low and high GnRH pulse frequencies respectively. We recently demonstrated that the transcriptional inhibitor, inducible cAMP early repressor (ICER), is stimulated at high GnRH pulse frequencies and attenuates FSH[beta] transcription to contribute to GnRH pulse frequency-dependent regulation of FSH. We hypothesized that distinct GnRH pulse frequency-dependent signalling pathways modulate the activity of ICER and thereby control FSH[beta] gene expression. Using the L[beta]T2 gonadotrope-derived cell line, studies with pharmacologic inhibitors suggested a primary role for MEKI/II signalling in mediating GnRH stimulation of ICER. GnRH stimulation of L[beta]T2 cells in static culture resulted in a time-dependent increase in ICER mRNA at 12 and 24 h, as measured by qRT-PCR. Pharmacological blockade of MEKI/II with two selective inhibitors, U0126 and PD325901, significantly attenuated GnRH-stimulated ICER induction in a dose-dependent fashion, whereas PKC (GF109203X), CamKII (KN-93) and PKA (H89) inhibitors had minimal effects. Similarly, MEKI/II inhibition attenuated GnRH induction of nuclear ICER protein as determined by Western blot analysis, confirming that the reduction in ICER mRNA is paralleled by a decrease in ICER protein levels. In perifusion studies, as shown previously, ICER expression was increased by pulsatile GnRH, with significantly greater increases at high (every 30 min) compared to low (every 120 min) pulse frequencies, conversely to effects on FSH[beta] expression. Importantly, MEKI/II inhibition with PD325901 in perifusion completely inhibited ICER induction at a high GnRH pulse frequency (every 30 min), whereas stimulation of ICER expression at low GnRH pulse frequency (every 120 min) was unaffected. In conclusion, blockade of MEKI/II signal transduction reduces GnRH induction of ICER in static culture. In perifusion, induction of ICER by pulsatile GnRH is selectively attenuated at high pulse frequencies, with no effect at low frequencies of pulsatile GnRH. These findings suggest that MEK signalling pathways play a key role in decoding pulsatile GnRH inputs in the gonadotrope, contributing to the induction of ICER by high frequency pulsatile GnRH to attenuate FSH[beta] induction.[br][br]Nothing to Disclose: IRT, NAC, SX, QZ, SZ, RSC, UBK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1611 72 721 SAT-643 PO17-01 Saturday 660 2012


661 ENDO12L_SAT-644 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) N-Terminal Truncation of the qGHRH(1-40) Targeting Domain of the Targeted Secretion Inhibitor (TSI) SXN101959 Ablates Its Ability To Activate the GHRH Receptor, Internalize into Cells and Cleave Vesicle-Associated Membrane Protein 3 (VAMP3) Elaine A Harper, Mark S Elliott, Julien Browne, Verity A Cadd Syntaxin Ltd, Abingdon, UK SXN101959 is a TSI, based on Botulinum neurotoxin serotype D (BoNT/D), which is designed to inhibit the secretion of growth hormone (GH) from pituitary somatotroph cells. SXN101959 contains the peptide ligand qGHRH(1-40), which targets the protein to the growth hormone releasing hormone receptor (GHRH-R), a class B G protein coupled receptor (GPCR). The mechanism of action of SXN101959 is hypothesised to be dependent on binding to and activation of the GHRH-R, agonist-driven receptor internalisation and cleavage of the VAMP family of SNARE proteins. We have previously shown that SXN101959 can activate the GHRH-R, internalise into cells expressing the receptor and cleave VAMP proteins. However, the exact requirement for receptor activation in this process has not been proven and therefore forms the basis of this study. There is literature precedence showing the requirement of the N-terminus of class B GPCR agonists for receptor activation but this has not been shown for the GHRH-R. Therefore, initial studies were performed to establish the effect of sequential N-terminal truncation of qGHRH(1-40) on receptor activation. This was assessed by measurement of intracellular cAMP accumulation in CHO-K1 and GH3 cells expressing recombinant human and rat GHRH-R, respectively. The importance of receptor activation for SXN101959 internalisation was established through the use of TSI with N-terminally truncated qGHRH(1-40) ligand. Internalised TSI was determined by high-content screening and SNARE cleavage by western blotting for VAMP3.[br]SXN101959 and qGHRH(1-40) were both full agonists at rat and human GHRH-R, evoking concentration-dependent increases in cAMP accumulation. Sequential N-terminal truncation of free peptide or TSI targeting domain resulted in an increasing loss of efficacy at the GHRH-R. Sequential truncation of the TSI targeting domain also significantly reduced the ability of the TSI to internalise. TSI with truncated qGHRH(1-40) targeting domains were no better at cleaving VAMP3 than a non targeted un-liganded light-chain of BoNT/D while the intact targeted TSI produced a potent and dose-dependent cleavage of VAMP3.[br]Activation of the GHRH receptor by SXN101959 is critical to it[apos]s mechanism of action because it is required to trigger receptor mediated internalisation into endosomes of target cells.[br][br]Nothing to Disclose: EAH, MSE, JB, VAC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1637 72 722 SAT-644 PO17-01 Saturday 661 2012


662 ENDO12L_SAT-645 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Effects of Gonadotropin-Releasing Hormone (GnRH) Metabolite GnRH-(1-5) on Phosphorylation of Epidermal Growth Factor Receptor in Ishikawa Cells Madelaine J Cho-Clark, Darwin O Larco, T John Wu Uniformed Services University of the Health Sciences, Bethesda, MD The decapeptide, GnRH, is known as a central regulator of mammalian reproduction by regulating the secretion of luteinizing hormone and follicle-stimulating hormone at the pituitary. In addition to its pituitary function, it also has an extra-pituitary functions that include behavioral effects as well as regulatory roles in peripheral tissues. For example, GnRH analogues are known to induce apoptosis in various types of tumors. Furthermore, there is emerging evidence regarding the significance of GnRH degradation mediated by the zinc metalloendopeptidase EC3.4.24.15 (EP24.15) to generate the metabolite GnRH-(1-5). Our laboratory has previously shown that in endometrial cells GnRH-(1-5) stimulates proliferation while the treatment with full-length GnRH inhibits this effect. To determine the mechanism mediating the proliferative effect of GnRH-(1-5), we investigated changes in the phosphorylation of the epidermal growth factor receptor (EGFR) since hyperactivation of this pathway is responsible for cell growth and proliferation of certain cancers. Our initial studies with Ishikawa cells, an endometrial cancer cell line, show that 100nM GnRH-(1-5) treatment increases the levels of phospho-EGFR by 3-fold (p[lt]0.05) at three different tyrosine sites (Y992, Y1045, Y1068). Furthermore, preliminary screenings using a high-throughput [beta]-Arrestin recruitment assay (DiscoverX, Fremont, CA) indicated GnRH-(1-5) to have an affinity for orphan receptor, G-protein coupled receptor 101 (GPR101). To link these findings, we down-regulated GPR101 expression (p[lt]0.05) and found that GnRH-(1-5)-induced phosphorylation of the EGFR at all three tyrosine sites was inhibited. Interestingly, GnRH-(1-5) increased EGF secretion by 10-fold (p[lt]0.05). In summary, our results suggest that GnRH-(1-5) stimulates EGF secretion and phosphorylates EGFR via GPR101 in Ishikawa cells.[br][br]Sources of Research Support: National Science Foundation Grant IOS-0544150 and IOS-1052288, and Department of Defense Grant RO95FN and RO85EU.[br][br]Nothing to Disclose: MJC-C, DOL, TJW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1933 72 723 SAT-645 PO17-01 Saturday 662 2012


663 ENDO12L_SAT-646 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Oxytocin Stimulated NFAT Transcriptional Activation in Human Myometrial Cells Jason NA Pont, Craig A McArdle, Andres Lopez Bernal Bristol University, Bristol, UK Oxytocin (OXT) is a peptide hormone which stimulates smooth muscle contraction of the myometrium during parturition. It binds to the OXT receptor on myometrial cells to initiate an intracellular signalling pathway culminating in intracellular calcium [Ca[sup]2+[/sup]][sub]i [/sub]accumulation and thus contraction (2). In some systems[sub] [/sub][Ca[sup]2+[/sup]][sub]i [/sub]elevation stimulates nuclear translocation of the transcription factor, nuclear factor of activated T cells (NFAT) (1), which is transcriptionally active in arterial smooth muscle (3). Moreover an inhibitor of the NFAT signalling pathway delayed the onset of parturition in mice by several hours (4). Therefore we have investigated the role of NFAT in the mechanism of action of OXT in human myometrial cells.[br]Here we have transduced myometrial cells with recombinant adenovirus expressing a NFATC1-EFP reporter and used a semi automated imaging system, as previously described (1), to monitor effects of OXT on reporter localisation in live cells. OXT induced nuclear translocation of NFATC1-EFP in a dose responsive and reversible manner; an effect which was inhibited by the OXT antagonists L368899 and FK506 and calcineurin inhibitors cyclosporine A and FK506. Pulsatile stimulation with OXT (5 mins) was mirrored by a cumulative nuclear translocation of NFATC1-EFP in a pulse frequency and amplitude dependent manner. The amplitude of NFATC1-EFP nuclear translocation with pulsatile OXT stimulation (every 30 mins), over 3 hr, was comparable to that with sustained stimulation, thus is a more efficient signalling mechanism within the cell; as [Ca[sup]2+[/sup]][sub]i [/sub]levels are only elevated for 5 min intervals rather than constantly throughout the time course. Endogenous NFAT was transcriptionally active in myometrial cells; as OXT induced activity of a NFAT-response element-luciferase reporter construct and stimulated calcineurin-NFAT dependent expression of [italic]RGS2, RCAN1 [/italic]and [italic]COX2 [/italic]mRNA. Moreover OXT dependent transcriptional activity was translation dependent, as cycloheximide abolished [italic]RGS2 [/italic]transcriptional activity. Indeed, cycloheximide actually increased OXT effects on [italic]RCAN1 [/italic]and [italic]COX2 [/italic]expression, demonstrating the existence of a novel negative feedback loop that suppresses OXT effects on gene expression and is dependent upon neosynthesis of undefined protein(s).[br]In conclusion this study identifies a novel signalling mechanism in the myometrium, whereby the calcineurin-NFAT signalling pathway mediates OXT dependent transcriptional activity.[br][br]1. Armstrong, S.P., et al., Pulsatile and sustained gonadotropin-releasing hormone (GnRH) receptor signaling: does the Ca2+/NFAT signaling pathway decode GnRH pulse frequency? J Biol Chem, 2009. 284(51): p. 35746-57. 2. Lopez Bernal, A., Mechanisms of labour--biochemical aspects. Bjog, 2003. 110 Suppl 20: p. 39-45. 3. Nilsson, L.M., et al., Novel blocker of NFAT activation inhibits IL-6 production in human myometrial arteries and reduces vascular smooth muscle cell proliferation. Am J Physiol Cell Physiol, 2007. 292(3): p. C1167-78. 4.Tabata, C., et al., Calcineurin/NFAT pathway: a novel regulator of parturition. Am J Reprod Immunol, 2009. 62(1): p. 44-50.[br][br]Sources of Research Support: BBSRC.[br][br]Nothing to Disclose: JNAP, CAM, ALB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 453 72 724 SAT-646 PO17-01 Saturday 663 2012


664 ENDO12L_SAT-647 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Luteinizing Hormone Receptor Signaling in MA-10 Mouse Leydig Cells Malena Rone, Vassilios Papadopoulos The Research Institute of the McGill University Health Centre, Montr[eacute]al, Canada The Leydig cells of the testis are responsible for the production of testosterone. Steroid production is initiated by the binding of lutenizing hormone (LH) to the G-Protein Coupled Receptor (GPCR) LH Receptor (LHR) located at the plasma membrane. The binding of LH to LHR results in the activation of the G-protein signaling cascade, the production of cAMP and resulting in the production of testosterone. After the binding of LH to LHR, the receptor is internalized to the endocytic pathway, a process that is proposed to terminate the ability of the LHR to produce cAMP and thus testosterone. This hypothesis was recently questioned as it was reported that the GPCR thyroid-stimulating hormone receptor continues to signal upon internalization(1). Therefore to determine if the LHR present in mouse MA-10 Leydig cell line functions similarly, we silenced the Vacuolar Protein Sorting34 (VPS34) a class III phosphoinositide 3-kinase which is proposed to function in the endosomal pathway. The decreased protein expression resulted in increased steroid production upon stimulation by human Chorionic Gonadotropin (hCG); a similar increase was not observed when the cells were stimulated with cAMP, a process which bypasses LHR signaling. To further determine if the LHR was signaling in the endosomal pathway, we stimulated control and VPS34 knockdown MA-10 cells with hCG for 15 minutes before acid washing (pH 3.0), removing any bound but not internalized hCG present on the plasma membrane. We then measured steroid production after two hours, in which the VSP34 siRNA-treated cells had significantly produced more steroids over both control and scrambled siRNA-treated cells. These results confirmed that the signaling of the LHR continues in the endosomal pathway and not only at the plasma membrane. To determine what mechanism of the endosomal trafficking pathway was altered we examined the role of Rab5 and Rab7, proteins known to function in the early to late endosomal transition. Using transient transfections of constructs of wild-type, dominant negative and constitutively active Rabs 5 [amp] 7 we did not observe alteration in steroid production, suggesting that VPS34 functions in another mechanism in endosomal trafficking. These initial results demonstrate that the LHR does continue to signal upon internalization from the plasma membrane, suggesting a mechanism of localized intracellular signaling controlling androgen formation in Leydig cells.[br][br](1) Calebiro D et al., PLoS Biol 2009; 7:e1000172.[br][br]Sources of Research Support: A grant from the Canadian Institutes of Health Research (MOP102647) and a Canada Research Chair in Biochemical Pharmacology to V.P. M.R. was supported by postdoctoral fellowships from the RI-MUHC. The Research Institute of MUHC was supported by a Center grant from Le Fonds de la recherche du Qu[eacute]bec-sant[eacute].[br][br]Nothing to Disclose: MR, VP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1560 72 725 SAT-647 PO17-01 Saturday 664 2012


665 ENDO12L_SAT-648 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Activation of AMPK Inhibits Luteinizing Hormone-Induced mTORC1 Signaling in Theca-Interstitial Cells Murugesan Palaniappan, KMJ Menon University of Michigan Medical School, Ann Arbor, MI Theca-interstitial (T-I) cells play a crucial role in ovarian physiology by synthesizing androgens, a vital precursor for estrogen synthesis by the granulosa cells. In the human, hyperactivity of T-I cell function is associated with conditions that lead to anovulatory hyperandrogenism, characteristic of polycystic ovarian syndrome (PCOS). Our previous studies have shown that LH/hCG activates mammalian target of rapamycin complex 1 (mTORC1) signaling leading to T-I cell proliferation. Activation of 5[apos]-adenosine monophosphate-activated protein kinase (AMPK) has been shown to inhibit cell proliferation. Furthermore, LH is known to reduce AMPK activation. Since mTORC1 and AMPK pathways exert opposite effects on cell proliferation, the present studies were undertaken to determine how these two pathways are integrated in response to LH stimulation. T-I cells were isolated from 25 day-old rat ovaries by collagenase digestion and cultured in the presence of hCG (50 ng/ml) for different time intervals. Whole cell lysates were analyzed for phosphorylations of AMPK and downstream targets of mTORC1, S6K1 and CREB by Western blot analysis. The results showed that treatment with hCG reduced AMPK phosphorylation in a time-dependent manner while producing a stimulatory effect on phosphorylation of downstream targets of mTORC1, S6K1 and CREB. In order to determine the effect of AMPK activation on hCG-induced S6K1 and CREB phosphorylation, T-I cells were pretreated with AICAR, a pharmacological activator of AMPK, for 1h followed by stimulation with hCG for 15 min. The cell lysates were analyzed for phosphorylation of S6K1 and CREB. The results revealed that while, as expected, hCG treatment elicited an increase in the phosphorylation of S6K1 and CREB, AICAR treatment prior to hCG stimulation completely blocked these responses suggesting that activation of AMPK inhibits hCG-induced mTORC1 signaling. To examine the mechanism of AMPK activation on T-I cell proliferation, T-I cells were pretreated with AICAR for 1 h followed by hCG-stimulation for additional 24 h, and total cell lysates were examined for cell proliferation markers by immunoblot analysis. The results showed that hCG treatment increased the expression of PCNA and cyclin D3, whereas AMPK activation abolished this hCG- induced PCNA and cyclin D3 protein expression. Taken together, these results show that activation of AMPK inhibits T-I cell proliferation by blocking mTORC1 signaling in T-I cells.[br][br]Sources of Research Support: NIH Grant HD-38424.[br][br]Nothing to Disclose: MP, KMJM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1962 72 726 SAT-648 PO17-01 Saturday 665 2012


666 ENDO12L_SAT-649 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Dimerization of the Luteinizing Hormone/Chorionic Gonadotrophin Receptor (LHCGR); Impact on Ligand Specificity and Intracellular Signaling Responses Kim C Jonas, Adolfo Rivero-Muller, Yen-Yin Chou, Tae H Ji, Aylin C Hanyaloglu, Ilpo T Huhtaniemi Imperial College, London, UK; University of Turku, Turku, Finland; University of Kentucky, Lexington, KY G protein-coupled receptors (GPCRs) are one of the largest families of the mammalian genome mediating the effects of 40% of prescription drugs. Accepted dogma states that GPCRs exist as monomers, however it is becoming increasingly clear that GPCRs can form higher order dimer/oligomer complexes that can diversify receptor function. The LHCGR, a Class A GPCR, primarily couples to the G[sub][alpha]s[/sub] pathway to mediate its reproductive functions. We have recently reported the first [italic]in vivo[/italic] evidence for the physiological importance of Class A GPCR homodimerisation; transgenic co-expression of binding deficient LHCGR (LHCGR[sup]-LH[/sup]) and signalling deficient LHCGR (LHCGR[sup]-cAMP[/sup]) could reverse the hypogonadism and infertility of male LHCGR null mice. Utilising the LHCGR[sup]-LH[/sup] and LHCGR[sup]-cAMP[/sup] as tools for studying the role of cis (same receptor binding hormone and propagating signal) and trans (one receptor partner binding ligand and one receptor partner propagating signal) activation through receptor dimerisation, we aimed to investigate the [italic]in vitro[/italic] downstream signalling pathways stimulated by these distinct modes of receptor activation. Furthermore, if cis and trans activation of LHCGR could unveil the potential for signal bias between the endogenous ligands of LHCGR, LH and hCG. In cell lines stably expressing either WT LHCGR or LHCGR[sup]-LH[/sup]/LHCGR[sup]-cAMP[/sup], treatment with either hCG or LH at 0.001nM to 100nM showed comparable maximum G[sub][alpha]s[/sub]- cAMP responses, indicating that trans activation was sufficient to mediate the G[sub][alpha]s[/sub] response to both ligands. Equivalent hCG-dependent IP[sub]1[/sub] and Ca[sup]2+[/sup] responses were observed in both WT and trans activational modes of signalling (P[gt]0.05). Interestingly, LH-dependent Ca[sup]2+[/sup] responses were severely diminished in the trans activational model (WT LHCGR 127.0[plusmn]9.6 vs LHCGR[sup]-LH[/sup]/LHCGR[sup]-cAMP[/sup] 72.6[plusmn]9.7, arbitary units, n=4, P[lt]0.05). Similar findings were observed with IP[sub]1[/sub] responses, revealing a requirement for cis activation for full LH-dependent[sub] [/sub]IP[sub]1[/sub] and Ca[sup]2+[/sup] responses. Conductance of a phospho-kinase array showed differences in signalling pathways activated in WT LHCGR and LHCGR[sup]-LH[/sup]/LHCGR[sup]-cAMP[/sup] cell lines, though only a few distinctions between ligands existed. Novel signalling targets of LHCGR were also identified. These results indicate that cis and trans modes of GPCR activation can result in diversification of ligand-activated signalling pathways and may provide insight into how ligand and receptor subtypes can facilitate multiple physiological functions.[br][br]Sources of Research Support: The Wellcome Trust, BBSRC and The Academy of Finland.[br][br]Nothing to Disclose: KCJ, AR-M, Y-YC, THJ, ACH, ITH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1913 72 727 SAT-649 PO17-01 Saturday 666 2012


667 ENDO12L_SAT-650 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) A Novel Antibody against the Transmembrane Domain of Glycoprotein Hormone Receptors Inhibits Hormone Actions and Suppresses Gain-of-Function Mutation without Affecting Hormone Binding Ritankar Majumdar, Reema S Railkar, Rajan R Dighe IISc, Bangalore, India The exoloops of glycoprotein hormone receptors transduce the signal generated by ligand-Ectodomain interactions to the transmembrane helices, either through direct hormonal contact and/or by modulating inter-domain interactions between the hinge region and the Transmembrane Domain (TMD). In this study, the ligand induced conformational alterations in the hinge regions and the inter-helical loops of LHR/FSHR/TSHR were mapped using exoloop specific polyclonal antibody generated against mini-Trans membrane domain (mini TMD) protein. This protein was designed to mimic the native exoloop conformations and created by joining the TSHR exoloops which were constrained by helical tethers and library derived linkers. The antibody against mini-TMD specifically recognized all three glycoprotein hormone receptors and inhibited basal and hormone stimulated cAMP production without affecting hormone binding. Interestingly, binding of the antibody to all three receptors was abolished with prior incubation of the receptors with the respective hormones suggesting that the exoloops are buried in the hormone-receptor complexes. The antibody also suppressed the high basal activities of gain-of-function mutations in the hinge regions, exoloops and TMDs such as those involved precocious puberty and thyroid toxic adenomas. This provides a novel therapeutic strategy to treat such conditions. Using the antibody and deletion or chimeric receptor mutants, we demonstrate that the hinge region and exoloops of LHR interact differently than those of FSHR and TSHR. Computational analysis suggests that miniTMD specific antibodies act by conformationally locking the transmembrane helices by restraining the exoloops and juxtamembrane regions. Using glycoprotein hormone receptors as model, we describe a novel computational approach of generating TMDs that can be used to produce interesting antibodies with therapeutic potential, as well as, explain the role of exoloops during receptor activation.[br][br]Sources of Research Support: Grants from the Department of Biotechnology, CSIR, and DST, Government of India, New Delhi, India.[br][br]Nothing to Disclose: RM, RSR, RRD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 482 72 728 SAT-650 PO17-01 Saturday 667 2012


668 ENDO12L_SAT-651 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Sustained G Protein Signaling Arising from a PTH Receptor[ndash]Arrestin[ndash]G-Protein Complex: A Non-Canonical Mode of GPCR Signaling Vanessa L Wehbi, Hilary P Stevenson, Timothy N Feinstein, Guillermo A Calero, Guillermo G Romero, Jean-Pierre Vilardaga University of Pittsburgh School of Medicine, Pittsburgh, PA; University of Pittsburgh School of Medicine, Pittsburgh, PA Influential studies on desensitization of activated [beta]2-adrenergic receptor and rhodopsin [ndash]two prototypical cell surface G protein-coupled receptors (GPCRs)[ndash] by arrestins led to what is currently thought to be a universal desensitization model for GPCR (1-5). Arrestins are known to terminate G protein-dependent signaling by preventing receptor and G protein coupling, and promoting receptor internalization via clathrin-coated pits (6,7). However, the parathyroid hormone type 1 receptor (PTHR), a medically essential GPCR that is critically important for bone and mineral ion metabolism, does not follow this conventional desensitization paradigm.[br][beta]-arrestins prolong Gs-mediated cAMP stimulation triggered by PTH, a process that correlates with the persistence of arrestin-PTHR complexes on endosomes, and which is thought to be associated with the prolonged physiological calcemic and phosphate responses of some PTH analogs (8-11). This presents an inescapable paradox for the arrestin-mediated receptor-G protein decoupling model: it is not possible that a receptor be both silenced by [beta]-arrestins and yet continue to generate cAMP after arrestin binding and arrestin-mediated internalization. Here we used biochemical and biophysical approaches to demonstrate how arrestins support sustained PTHR signaling. We found that PTHR formed a ternary complex with arrestin and the G[beta][gamma] dimer in response to PTH stimulation, which in turn prolonged receptor signaling by mechanisms that permitted multiple rounds of G-protein coupling and activation. This work provides a new model of GPCR signaling prolonging the hormone effect on the receptor.[br][br](1) Wilden, U. et al. Proc Natl Acad Sci 1986. (2) Lohse, M. J. et al. Science 1990. (3) Lefkowitz, R. J., J Biol Chem 1998. (4) Gurevich, V. V.et al., (ed J. Giraldo, and Pin, J.-P.) Ch. 17, 335-355 (Royal Society of. Chemistry, 2011). (5) Lohse, M. J. et al. J Biol Chem 1992. (6) Krupnick, J. G. et al., J Biol Chem 1997. (7) Ferguson, S. S. et al. Science 1996. (8) Ferrandon, S. et al., Nat Chem Biol 2011. (9) Feinstein, T. N. et al. Nat Chem Biol 2011. (10) Castro, M. et al. Endocrinology 2002. (11) Okazaki, M. et al. Proc Natl Acad Sci U 2008.[br][br]Sources of Research Support: This work was supported by the National Institutes of Health (NIH). award R01DK087688 (to J.-P.V.).[br][br]Nothing to Disclose: VLW, HPS, TNF, GAC, GGR, J-PV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1926 72 729 SAT-651 PO17-01 Saturday 668 2012


669 ENDO12L_SAT-652 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Identification of Coupling Specificity between Somatostatin Receptor 5 (SST5) and G Proteins by a Bioluminescence Resonance Energy Transfer (BRET) Technique: The Role of GoA Protein Erika Peverelli, Giovanna Mantovani, Eleonora Vitali, Andrea G Lania, Paolo Beck-Peccoz, Anna Spada Fondazione IRCCS Ca[apos] Granda Ospedale Maggiore Policlinico, University of Milan, Milan, Italy; Istituto Clinico Humanitas IRCCS, University of Milan, Milan, Italy In the present study we employed a novel bioluminescence resonance energy transfer (BRET) biosensor to study the coupling specificity of somatostatin receptor 5 (SST5) and its naturally occurring mutant R240W in living cells. Previous data from our laboratory demonstrated in a rat pituitary cell model that SST5 carrying the R240W mutation as well as other mutations in the third intracellular loop maintained the ability to inhibit intracellular cAMP levels similarly to the wild-type but failed to mediate the inhibition of intracellular calcium levels, GH release and cell proliferation, suggesting that different regions of SST5 are required for the activation of different signaling pathways. The BRET biosensor may monitor the activation of G proteins in response to SST5 activation by the specific agonist BIM23206. The energy transfer occurs within two subunits of the heterotrimeric G protein complex, the energy donor (G alpha subunit fused to Renilla Luciferase) and the acceptor (G gamma2 subunit fused to GFP10). G protein activation induces a structural rearrangement that increases the distance between the donor and the acceptor with a consequent decrease in the energy transfer. To detect specific G protein activation in living cells, we expressed in HEK293 cells wild-type or mutant human SST5 together with different G alpha protein subunits (i1, i2, i3, oA, oB or q). We demonstrated that wild-type SST5 activated Gi (1, 2 and 3) and Go (A and B), whereas R240W SST5 maintained the ability to activate all Gi and GoB, but not GoA. As expected, neither the wild-type nor mutant receptors activated Gq. To investigate the role of GoA protein in SST5-mediated signal transduction in a pituitary cell model, we cotransfected cultured cells from GH-secreting adenomas with SST5 and a pertussis toxin (PTX)-resistant GoA protein. In PTX-treated cells, GoA restored the ability of BIM23206 to inhibit ERK1/2 phosphorylation and GH secretion. In conclusion, our data first demonstrated the coupling specificity of SST5 in living cells and revealed a crucial role for GoA protein in SST5 signaling in GH-secreting adenoma cells.[br][br]Sources of Research Support: Grant from AIRC (Associazione Italiana per la Ricerca sul Cancro) to G.M.[br][br]Nothing to Disclose: EP, GM, EV, AGL, PB-P, AS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1496 72 730 SAT-652 PO17-01 Saturday 669 2012


670 ENDO12L_SAT-653 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Overexpression of Human Somatostatin Receptor-3 Mediates Diverse Antiproliferative Signaling in MCF-7 and MDA-MB-231 Breast Cancer Cells Sajad A War, Ujendra Kumar The University of British Columbia, Vancouver, Canada Somatostatin (SST) is a pleiotropic inhibitory peptide and regulates cell proliferation through five different receptor subtypes coupled to G[sub]i[/sub] proteins namely somatostatin receptors (SSTR1-5). The inhibition of cell proliferation by SSTR subtypes engages multiple converging mechanisms including the activation of cytostatic and pro-apoptotic pathways. SSTR3 is structurally different from other SSTRs in possessing the longest C-tail containing 100 amino acids. This difference might account for its unique ability in activating the apoptotic cascades. In spite of significant antiproliferative properties [italic]in vitro[/italic], SST analogs exhibit, at best, modest antitumor activity in several cancers. This discrepancy has been largely attributed to the diminished SSTR expression. The objective of the present study was to identify the putative antiproliferative mechanisms of overexpressed SSTR3 in estrogen receptor-alpha (ER[alpha]) +ve MCF-7 (R3-MCF-7) and ER[alpha] -ve MDA-MB-231 (R3-MB-231) breast cancer cells. Our immunocytochemistry and Western blot analysis data showed an up-regulation of SSTR3 at the cell surface in response to SSTR3-selective agonist in R3-MCF-7, whereas, receptor internalization was observed in R3-MB-231 suggesting a possible role for ER[alpha] in SSTR3 trafficking. MTT assay in R3-MCF-7 and R3-MB-231, demonstrated severely blunted response on cell proliferation by epidermal growth factor, in addition to the enhanced antiproliferative spectrum of SSTR3-specific agonist, when compared to MCF-7 and MDA-MB-231, respectively. Most importantly, the antiproliferation in R3-MCF-7 correlated with increased PARP-1 activation and TUNEL staining, suggesting apoptosis even under basal conditions, whereas the induction of cyclin-dependent kinase inhibitor p27[sup]Kip1 [/sup]predicted a cytostatic effect. On the other hand, R3-MB-231 mediated antiproliferation was largely associated with cell cycle arrest as evident by increased expression of p27[sup]Kip1[/sup]. Conversely, no activation of PARP-1 and TUNEL-positive reaction in R3-MB-231 might provide an indirect evidence for the potential role of ER[alpha] as a preferred partner in SSTR3 mediated apoptosis in R3-MCF-7. Taken together, these findings suggest that R3-MCF-7 cells propagate the apoptotic and cytostatic functions of SSTR3, whereas in R3-MB-231 cells, it is predominantly cytostatic. In conclusion, these findings provide new insights in understanding the molecular interactions involving SSTRs, specifically SSTR3 in tumor biology.[br][br]Sources of Research Support: CIHR, CBCF-BC/Yukon, and MSFHR.[br][br]Nothing to Disclose: SAW, UK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 174 72 731 SAT-653 PO17-01 Saturday 670 2012


671 ENDO12L_SAT-654 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Somatostatin Receptor-2 Mediated Regulation of Hypertrophy and Adrenergic Receptor Functions in H9c2 Cells Rishi K Somvanshi, Shenglong Zou, Xiaofan Qiu, Ujendra Kumar The University of British Columbia, Vancouver, Canada Somatostatin receptors (SSTRs) and adrenergic receptors (ARs) are the members of G-Protein coupled receptor family and via coupling to distinct G-proteins demonstrate great diversity in physiological response of the cells. The role of [beta]-ARs in hypertrophy and cardiac complications is well understood. We recently demonstrated that SST plays critical role in the regression of hypertrophy in H9c2 cells. However, which SSTR subtype mediates this effect of SST is largely elusive. In the present study, we explored the effect of SSTR2 subtype on signaling pathways associated with hypertrophy following [beta][sub]1[/sub]AR and [beta][sub]2[/sub]AR activation in H9c2 cells. First, we determined the colocalization and interaction of SSTR2 with [beta]-ARs in H9c2 cells using immunocytochemistry and co-immunoprecipitation. We also investigated the effects of SSTR2 on the hypertrophic responses and associated signaling pathways including cAMP/PKA, PKC, ERK1/2 and NFAT in H9c2 cells in presence or absence of [beta]-ARs activation. Our results demonstrate that SSTR2 and [beta][sub]1[/sub]AR or [beta][sub]2[/sub]AR are well expressed in H9c2 cells and exhibit colocalization. Isoproterenol mediated increase in cell size and protein content (an index of hypertrophy) was significantly abolished in presence of SSTR2 specific agonist L-779,976. Forskolin stimulated cAMP was inhibited in presence of SSTR2 agonist in concentration dependent manner. Isoproterenol mediated PKA phosphorylation was enhanced in the presence of SSTR2 specific agonist. Conversely, no discernable changes in PKA phosphorylation were observed with formoterol with or without SSTR2 agonist. SSTR2 agonist L-779,976 in combination with isoproterenol or formoterol increases phospho-ERK1/2 and phospho-NFAT levels whereas inhibits PKC[alpha] and PKC[beta] expression in concentration dependent manner. These data demonstrate that [beta][sub]1[/sub]AR and [beta][sub]2[/sub]AR stimulated signaling is modulated upon SSTR2 activation. Results presented here provide direct evidence for the role of SSTR2 in modulation of signaling pathways attributed to cardiac hypertrophy using H9c2 cells as a model.[br][br]Sources of Research Support: Canadian Institute of Health Research Grants (MOP 10268 and MOP 74465), grant from Canadian Breast Cancer Foundation BC/Yukon and grant from NSERC, Canada to UK. UK is a Senior Scholar of Michael Smith Foundation for Health Research.[br][br]Nothing to Disclose: RKS, SZ, XQ, UK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 101 72 732 SAT-654 PO17-01 Saturday 671 2012


672 ENDO12L_SAT-655 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Agonist-Induced Internalization and Desensitization of the Apelin Receptor George R Pope, Craig A McArdle, Steve J Lolait, Anne-Marie O[apos]Carroll University of Bristol, Bristol, UK The apelin receptor (APJ) is a G protein-coupled receptor (GPCR) that binds the endogenous ligand apelin, and is linked via G[alpha]i or G[alpha]q to a network of intracellular pathways that include activation of extracellular-regulated kinases (ERK) [1-3]. APJ has roles in many physiological systems, with effects on angiogenesis, drinking behaviour and blood pressure [4-6]. GPCRs can be induced to undergo internalization in the presence of agonists and, upon binding of apelin-13, APJ has been shown to internalize via a clathrin-coated vesicle dependent mechanism with recruitment of [beta]-arrestin 1[amp]2, and rapid receptor recycling to the cell surface [7]. Little is known however regarding the regulation of APJ function through ligand-induced internalization and responsiveness.[br]In this study we explore apelin-induced APJ internalization and down-regulation using automated imaging to monitor expression of hemagglutinin-tagged mouse APJ, and dominant negative mutants (DNMs) of proteins involved in clathrin-mediated endocytosis. We also explore the mechanisms underlying apelin-induced APJ desensitization, measured using an ERK phosphorylation assay.[br]We show that incubation with apelin-13 down-regulates cell surface and whole cell levels of APJ, and induces receptor internalization, followed by rapid recycling of the receptor. Using DNMs we show internalization of APJ to be dependent upon dynamin, GRK2 and EPS15, indicating that each of these proteins plays a key role in this process. No dependence upon [beta]-arrestin 1 is seen. Additionally, while we show apelin stimulation of the ERK pathway at 1-h following initial apelin incubation, this is followed by a period (up to 24-h) of desensitization. EGF, adrenaline and nicotinic acid which activate ERK via Ras/Raf, G[alpha]q and G[alpha]i pathways respectively, are capable of stimulating phosphorylation of ERK at all time points, indicating that the lack of ERK activation is not due to changes in the ability of ERK to be phosphorylated. Therefore, the initial exposure to apelin leads to a time-dependent reduction in ERK activation specifically to a second exposure to apelin. This reduction in ERK activity is significantly restored after transfection with DNMs.[br]In conclusion, we show that agonist-induced APJ internalization and subsequent down-regulation may have a role to play in desensitization of apelin-mediated ERK activation.[br][br][1] Tatemoto K et al., Biochemical and Biophysical Reaearch Communications 1998; 251:471. [2] Szokodi I et al., Circulation Research 2002; 91:434. [3] Masri B et al., Biochemical and Biophysical Reaearch Communications 2002; 290:539. [4] Kasai A et al., Biochemical and Biophysical Reaearch Communications 2004; 325:395. [5] Roberts EM et al., The Journal of Endocrinology 2009; 202:453. [6] Ishida J et al., The Journal of Biological Chemistry 2004; 279:26274. [7] Evans NA et al., Journal of Neurochemistry 2001; 77:476.[br][br]Sources of Research Support: The Biotechnology and Biological Sciences Research Council (BBSRC) PhD Studentship SG1523.[br][br]Nothing to Disclose: GRP, CAM, SJL, A-MO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 720 72 733 SAT-655 PO17-01 Saturday 672 2012


673 ENDO12L_SAT-656 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Protein Kinase A Type-II Activity Is Altered in the Mouse Model of Fibrous Dysplasia Eva Szarek, Maria Nesterova, Jean B Regard, Yingzi Yang, Constantine A Stratakis Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, MD; Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, MD Fibrous dysplasia (FD), a skeletal dysplasia affecting bone marrow stromal cells, results from a mosaic gain-of-function mutation in GNAS, causing constitutive activation of G[alpha]s, a member of the G[alpha] protein family through which G-protein coupled receptors (GPCRs) signal. FD is characterized by extensive marrow fibrosis associated with increased proliferation, reduced osteoblastic cell differentiation and poorly mineralized bone. It remains unclear what molecular mechanisms involved in G[alpha]s activation in FD. It is known that GPCRs signal through G[alpha]s-coupled subunits whereby they transduce signals to adenylyl cyclase (AC), converting ATP to cAMP, in turn activating PKA, a holoenzyme consisting of a homodimer of regulatory subunits (R1[alpha],R1[beta],RII[alpha], [amp] RII[beta]) and two inactive catalytic subunits (C[alpha],C[beta],C[gamma] and Prkx), each catalytic subunit bound to one regulatory subunit of the dimer. It is important to note: R1[alpha] and R1[beta] form the PKA type I isozyme(PKA-I), whereas RII[alpha] and RII[beta] form PKA type II(PKA-II); these isozymes have different cellular localizations, functions, and affinity to cAMP. Activation of PKA occurs through parathyroid hormone(PTH) or PTH-related protein and AC-dependent cAMP generation. cAMP signaling is essential for normal bone development. To examine PKA signaling through G[alpha]s in FD we utilized a FD-like mouse model whereby G[alpha]s was conditionally removed from the developing limb mesenchyme with the [italic]Prx1-Cre[/italic] line. Because these mice died postnatally we examined mice initially at embryonic day € 10 and E18. We first examined PKA and phosphodiesterase (PDE) activities. Loss of G[alpha]s in the developing limb mesenchyme did not result in significant decrease in cAMP-stimulated kinase activity. We next measured PDE activity to determine whether cAMP levels were the result of PDE activity. Total PDE activity was significantly increased (p=0.005) in the developing limb mesenchyme of G[alpha]s mutants. At E18 we could not see a significant difference in cAMP-stimulated kinase activity, likely due to the surrounding tissue on the limb. By Western blot analysis, we determined that RII[alpha] and RII[beta] (PKA-II) were expressed at low levels. These results are in line with that seen in human mesenchymal cells with decreased GNAS expression whereby there is an associated PKA-II activity decrease and a more mature-osteoblast-like phenotype[1]. Our data suggest that activated G[alpha]s proteins are important during skeletal development and disease by signaling through PKA-II isozymes.[br][br]1. Lietman, S.A., et al., [italic]Reduction in Gsalpha induces osteogenic differentiation in human mesenchymal stem cells[/italic]. Clinical orthopaedics and related research, 2005(434): p. 231-8.[br][br]Nothing to Disclose: ES, MN, JBR, YY, CAS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2164 72 734 SAT-656 PO17-01 Saturday 673 2012


674 ENDO12L_SAT-657 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) GLP-1-Mediated CREB Activation in ZDF Rat Brain and in Human Neural Stem Cell-Derived Neurons Kalpana Velmurugan, Subbiah Pugazhenthi University of Colorado Denver, Aurora, CO; Denver VA Medical Center, Denver, CO Insulin signaling plays a significant role in cerebral glucose metabolism and synaptic function in the brain. Cognitive dysfunction and decrease in brain size have been observed in patients with diabetes. Pathogenic pathways in the diabetic brain resemble early events in Alzheimer[apos]s disease (AD). Epidemiological studies have indeed identified diabetes as a risk factor for AD. Cognitive aging among diabetic patients will be a major health problem because of increasing longevity and intellectually challenging lifestyles. In this context, glucagon-like peptide-1 (GLP-1) has generated therapeutic interest because of its neuroprotective and antiinflammatory actions. GLP-1, released from the gut in response to food, increases insulin secretion by beta cells and decreases beta cell apoptosis. Although studies have reported GLP-1 actions in brain, limited information is available on its molecular mechanism of action, especially at the level of transcription factors. Cyclic AMP response element binding protein (CREB) is a nuclear transcription factor needed for cognition and memory formation. We have characterized the mechanism of CREB downregulation by oxidative stress and cytokines in beta cells of diabetic pancreas and in neurons of Alzheimer[apos]s brain.[br]In the current study, we examined GLP-1 action in the brain of ZDF rats, a model for type 2 diabetes and in differentiated human neural stem cells. We observed in ZDF rat hippocampal samples (i) decreased activation of Akt and CREB (ii) decreased expression of BDNF, a target of CREB and a neurotrophic growth factor and (iii) markers of inflammation. All these defects were significantly normalized by treatment with alogliptin, a DPP-4 inhibitor that sustains endogenous GLP-1 levels. Exendin-4, a GLP-1 analog, activated CREB, induced BDNF promoter and decreased expression of inflammatory genes in cultured human brain neurons. Exendin-4 protected these neurons from apoptosis induced by neurotoxins including Abeta, 4-hydroxynonenal and cytokines. The antiapoptotic effect of exendin-4 was significantly reduced following transduction with adenoviral dominant negative mutant for of CREB. GLP-1 can cross the blood brain barrier, thereby has a distinct advantage over several neurotrophic growth factors. Thus the dual actions of GLP-1 in beta cells of pancreas and in neurons of brain could be an effective therapeutic strategy to prevent cognitive decline and progressive neurodegeneration in aging population with diabetes.[br][br]Sources of Research Support: Takeda investigator-initiated Research Study.[br][br]Nothing to Disclose: KV, SP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2058 72 735 SAT-657 PO17-01 Saturday 674 2012


675 ENDO12L_SAT-658 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Phosphoinositide 3-OH Kinase Beta Mediates G-Protein Coupled Receptor-Dependent Protein Kinase D Activation Christine M Lynch, Ronald W Matheny United States Army Research Institute of Environmental Medicine, Natick, MA Phosphoinositide 3-OH kinase beta (p110 beta) is a lipid kinase associated with G-protein coupled receptor (GPCR) signal transduction in multiple tissues; however, its role as a signaling molecule has yet to be fully defined, particularly in skeletal muscle. To address this, we utilized an RNAi approach in C2C12 myoblasts and investigated whether p110 beta mediated GPCR-stimulated protein kinase D (PKD) activation. PKD was chosen as a readout of GPCR signaling due to previous observations demonstrating its activation in response to serum in a pertussis toxin-sensitive manner. We found that serum promoted PKD phosphorylation at Ser [sup]744/748[/sup] and Ser [sup]916[/sup] as well as PKD substrates possessing the PKD consensus phosphorylation motif; however, this effect was attenuated in cells lacking p110 beta. Conversely, loss of p110 beta did not affect stimulation of PKD phosphorylation by the phorbol ester TPA (12-O-Tetradecanoylphorbol-13-Acetate), suggesting that p110 beta lies upstream of protein kinase C (PKC) in the pathway for PKD activation. Serum contains multiple active components, and one such component is lysophosphatidic acid (LPA). Because LPA has been previously shown to activate PKD in non-muscle cells, we next tested: 1) whether LPA activates PKD in C2C12 myoblasts; and 2) whether p110 beta mediates LPA-induced PKD phosphorylation. We found that LPA stimulated PKD phosphorylation at Ser [sup]744/748[/sup] and Ser [sup]916[/sup]; and that this phosphorylation was attenuated in cells deficient in p110 beta. Taken together, these results suggest that serum- and LPA-stimulated PKD phosphorylation is mediated by p110 beta in myoblasts; moreover, these findings reveal a novel role for p110 beta upstream of PKC/PKD signaling.[br][br]Sources of Research Support: This work was supported by an appointment to the postgraduate research participation program at the United States Research Institute of Environmental Medicine administered by the Oak Ridge Institute for Science and Education through an interagency agreement between the United States Department of Energy and Unites States Army Medical Research and Materiel Command (C.M.L).[br][br]Nothing to Disclose: CML, RWM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1944 72 736 SAT-658 PO17-01 Saturday 675 2012


676 ENDO12L_SAT-659 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Functions of Transmembrane Domain 3 of Human Melanocortin-4 Receptor Xiu-Lei Mo, Rui Yang, Ya-Xiong Tao Auburn University, Auburn, AL The melanocortin-4 receptor (MC4R) is a G protein-coupled receptor that plays an essential role in regulating energy homeostasis. Defects in MC4R are the most common monogenic form of obesity, with more than 150 distinct MC4R mutations identified in human. A few studies suggested that transmembrane domain 3 (TM3) of MC4R is important for ligand binding and activation. Herein, we systematically analyzed functions of 26 residues in TM3 (excluding S136 and DRYFTI motif) using alanine-scanning mutagenesis. HEK293T cells were transiently transfected with wild type (WT) or mutant MC4Rs, and stimulated with the superpotent ligand NDP-MSH. Cell surface expression, ligand binding and signaling properties of these mutants were assessed. We showed that all 26 mutants had normal cell surface expression. Two mutants, D126A and L140A, were significantly defective in both ligand binding and cAMP signaling. V128A and I143A had normal ligand binding but impaired cAMP production. L140A had increased basal cAMP level (8-fold higher than that of WT). To further characterize the function of L140, we generated seventeen L140 mutants. We showed that fifteen L140 mutants had significantly decreased cell surface expression, with L140R and L140V expressed normally. Mutations of L140 to C, D, E, K, N, P, Q, S, T and V, cause constitutive activation in cAMP pathway. Four mutants (L140G, L140I, L140M and L140R) were signaling impaired in response to NDP-MSH stimulation. We further characterized the activation of ERK1/2 in constitutively active mutants (CAM) and signal defective mutants (SDM). We showed that five CAMs (L140K, L140N, L140P, L140T, and L140V) constitutively activated pERK1/2, and two SDMs (L140I and L140M) were impaired in pERK1/2 activation, similar to cAMP signaling. However, four CAMs (L140C, L140E, L140Q and L140S) had similar level of basal pERK1/2 as that of WT, and two SDMs (L140G and L140R) had similar level of ERK1/2 phosphorylation as that of WT upon NDP-MSH stimulation, different from their cAMP signaling properties, suggesting biased signaling in these mutant receptors. In summary, our studies demonstrate that residues D126, V128, L140 and I143 in TM3 are essential for MC4R ligand binding and/or signaling. Moreover, L140 is critical for locking MC4R in inactive conformation and several mutants at this locus display biased signaling in cAMP and ERK1/2 signaling pathways.[br][br]Sources of Research Support: NIH Grant R15DK077213 and Animal Health and Disease Research from Auburn University College of Veterinary Medicine awarded to YXT.[br][br]Nothing to Disclose: X-LM, RY, Y-XT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 314 72 737 SAT-659 PO17-01 Saturday 676 2012


677 ENDO12L_SAT-660 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Pharmacological Study of the Transmembrane Domain 6 of Human Melanocortin-4 Receptor Hui Huang, Ya-Xiong Tao Auburn University, Auburn, AL The melanocortin-4 receptor (MC4R) is a G protein-coupled receptor (GPCR) primarily expressed in the central nervous system. It is a critical regulator of energy homeostasis, regulating both food intake and energy expenditure. Mutations of MC4R gene have been identified as the most common cause of monogenic obesity. Therefore the MC4R has emerged as a premier target for obesity treatment. To gain a better understanding of the structure-function relationship of the MC4R, we sought to determine the function of each residue in transmembrane domain 6 (TM6) of MC4R using alanine-scanning mutagenesis. We generated thirty-one mutants and used the two endogenous ligands [alpha]-melanocyte stimulating hormone (MSH) and [beta]-MSH for ligand binding and signaling studies. We found that three mutants (H264A, L265A and Y268A) were severely defective in ligand binding, and four mutants (L250A, C257A, P260A and F261A) had decreased maximal binding. Seven mutants (K242A, L247A, I251A, W258A, P260A, F261A and F262A) displayed significant signaling impairments to [alpha]-MSH stimulation, and four additional mutants (T246A, L250A, G252A and F254A) were defective in cAMP production in response to [beta]-MSH stimulation. H264A had no detectable binding and showed no response to either [alpha]- or [beta]-MSH stimulation. The cell surface expression levels of H264A and L265A were not different from that of the WT MC4R while the cell surface expression of P260A was decreased. Nine mutants (K242A, G243A, T246A, L247A, I251A, G252A, W258A, P260A and H264A) had decreased basal activities. Seven mutants (M241A, A244G, L250A, A259G, I266A, F267A and I269A) were constitutively active and their basal activities could be inhibited by two MC4R inverse agonists, Ipsen 5i and ML00253764, with maximal inhibition ranging from 40% to 88%. Five of these mutants (M241A, L250A, I266A, F267A, I269A) were also constitutively active in the MAPK pathway with enhanced basal ERK phosphorylation. A244G and A259G did not have increased basal ERK phosphorylation. In summary, we identified residues that are important for cell surface expression, ligand binding, signaling, and constraining the wild type receptor in inactive conformation. We also reported constitutive activation of MAPK pathway. These data provided detailed information regarding the structure-function relationship of the TM6 of MC4R in receptor function.[br][br]Sources of Research Support: NIH Grant R15DK077213 and Animal Health and Disease Research from Auburn University College of Veterinary Medicine Awarded to YXT.[br][br]Nothing to Disclose: HH, Y-XT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 308 72 738 SAT-660 PO17-01 Saturday 677 2012


678 ENDO12L_SAT-661 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Mapping Structural Determinants within CRH-R1 3rd Intracellular Loop Important for G-Protein[ndash]Dependent and Independent ERK1/2 and p38MAPK Activation Anu Punn, Jing Chen, Hendrik Lehnert, Michael Levine, Dimitris Grammatopoulos University of Warwick, Coventry, UK; University of Luebeck, Luebeck, Germany; University of Pennsylvania School of Medicine, Philadelphia, PA Corticotropin releasing hormone (CRH) and urocortins (Ucn) activate two types of GPCRs, R1 and R2. The CRH-R1a signaling to intracellular effectors involves both G-protein dependent and independent pathways (1). To gain insights on the structural determinants of CRH-R1a signaling specificity we investigated the role of the juxtamembrane regions of CRH-R1a 3rd intracellular loop (IC3) by mutating specific residues to alanine (2). Basic residues were identified as important residues for regulating and limiting activation of adenylyl cyclase, through interactions with pertussis toxin-sensitive G-proteins. Moreover, basic residues within both juxtamembrane regions as well as aliphatic amino acids such as I293 and L294 within the N-terminus of IC3, were identified as important for ERK1/2 phosphorylation through activation of Gi- as well as Gq proteins. We investigated the effects of these individual mutations on signaling cross-talk and G-protein independent mechanisms involved in CRH-R1[alpha]- mediated ERK1/2 and p38 MAPK activation. Results showed that substitution of I293 and L294 but not R292 significantly impaired the ability of Ucn1 to activate ERK1/2 and p38 MAPK (50-70% reduction of maximal response). For mutant receptors such as R292A and K311A that retained Gq-coupling activity, residual MAPK responses to Ucn1 stimulation were sensitive to the activity of the cAMP-PKA-AKAP pathway. Treatment of cells with specific adenylyl cyclase and PKA inhibitors or Ht-31 to interrupt PKA binding to AKAP, increased Ucn1-induced ERK1/2 responses by 2-3 fold. Moreover, we analysed mutant receptor beta-arrestin recruitment and internalization characteristics; no differences were observed between wild type and individual mutant CRH-R1a in beta-arrestin plasma membrane translocation or receptor endocytosis suggesting that the diminished MAPK signalling was not due to impaired internalization characteristics. Interestingly, tandem mutations of the aminoacid cassette K311-K314 at the C-terminal segment altered agonist-induced receptor-beta-arrestin interaction and rate of receptor endocytosis. Indeed, the CRH-R1a(C-4A) receptor retained significantly more cAMP responsiveness after Ucn1 pretreatment, suggesting a higher degree of resistance to beta-arrestin- homologous desensitization. These data define a critical role for key amino acids within the hydrophobic N- and C-terminal microdomains of IC3 in the control of CRH-R1 signaling activity and beta-arrestin interactions.[br][br]1. Hillhouse EW, Grammatopoulos DK. Endocr Rev. 2006; 27:260-86. 2. Punn, et al, J Bio Chem, 2012 in press.[br][br]Sources of Research Support: Wellcome Trust.[br][br]Nothing to Disclose: AP, JC, HL, ML, DG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1651 72 739 SAT-661 PO17-01 Saturday 678 2012


679 ENDO12L_SAT-662 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Brain-Specific Angiogenesis Inhibitor 2 (BAI2) Intracellular Splice Variants Do Not Exhibit Specific Functional Properties Alexandra Kiess, Rainer Strotmann, Stine Holland, Torsten Schoneberg University of Leipzig, Leipzig, Germany Background: Brain-specific angiogenesis inhibitor 2 (BAI2) is a member of the adhesion-G protein-coupled receptors (GPCRs) superfamily with a dominant expression in brain. The putative ligands and functions of BAI2 are still unclear. The intracellular domain of BAI2 shows splice variants with unknown function.[br]Objective: In this study we analyzed the functional relevance of splice variants in the third intracellular loop (ICL3) and in the C terminus of BAI2.[br]Material and Methods: Ortholog sequences of ICL3 and the C terminus of BAI2 from different species were used to study their evolutionary conservation. The splice variants were tested for changes in receptor protein[apos]s membrane topology and for possible intra- and intermolecular protein interactions using different peptide array and ELISA techniques.[br]Results: The splice variants in the ICL3 were found to be highly conserved between species. The resulting protein variants did not change the topology of BAI2 in the plasma membrane and were not relevant for intramolecular interactions. The C-terminal human splice variant was also found in other primates and dogs, but not in mouse and rat. The human C-terminal splice variant showed high affinity to calmodulin and was not calcium-dependent.[br]Conclusion: Splice variants ICL3 and C terminus are a stable evolutionary feature of BAI2 implicating specific intracellular functions and interactions. The splice variants do not influence the receptor[apos]s membrane topology but may be involved in splice variant-specific recruiting of cytosolic proteins such as calmodulin.[br][br]Nothing to Disclose: AK, RS, SH, TS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 261 72 740 SAT-662 PO17-01 Saturday 679 2012


680 ENDO12L_SAT-663 POSTER SESSION: G Protein-Coupled Receptor Biology (1:30 PM-3:30 PM) Plasma Membrane Abundance of Wild-Type Human Calcium-Sensing Receptor Homodimers Are the Key Regulators of Calcium-Sensing in FHH Patients Michael P Grant, Ann Stepanchick, Gerda E Breitwieser Geisinger Clinic, Danville, PA The primary regulator of parathyroid hormone is the extracellular calcium-sensing receptor (CaSR), which rapidly responds to small changes in circulating calcium. Loss-of-function mutations in CaSR cause familial hypocalciuric hypercalcemia (FHH) or neonatal severe hyperparathyroidism (NSHPT), when one or both alleles are affected, respectively. Gain-of-function mutations cause autosomal dominant hypocalcemia. The majority of CaSR mutations are loss-of-function, having a rightward shift in their [Ca[sup]2+[/sup]][sub]o [/sub]dose-response relationship. CaSR functions as a dimer. Heterodimerization between [italic]wt[/italic] and mutant CaSR occurs and may contribute to the benign phenotype in FHH. We recently demonstrated that CaSR signaling is regulated by a novel mechanism which relies on agonist-mediated CaSR plasma membrane insertion coupled to constitutive internalization to control signaling amplitude ([bold]A[/bold]gonist-[bold]D[/bold]riven [bold]I[/bold]nsertional [bold]S[/bold]ignaling, ADIS). In the present study, we used similar approaches (TIRF microscopy, calcium imaging and co-immunoprecipitations) to determine the mechanism(s) contributing to the FHH phenotype. Expression of loss-of-function mutants (R227Q, E297K, C395R, R795W, P798T and A804D) alone showed the expected severe phenotype, with absence of signaling and agonist-evoked trafficking to the plasma membrane. Coexpression of mutants with [italic]wt[/italic]CaSR (1:1 ratio of cDNAs), however, resulted in significant rescue of both net signaling and specific trafficking of mutant CaSRs to the plasma membrane. We then tested the hypothesis that rescue of mutant trafficking was a result of signaling by [italic]wt[/italic]CaSR. Expression of varying ratios of cDNAs for wt and loss-of-function mutants (1:4 or 4:1 ratios), caused rightward or leftward shifts in the EC[sub]50[/sub] for mutant CaSR trafficking as a function of [italic]wt[/italic]CaSR signaling. Results suggest that plasma membrane abundance of [italic]wt[/italic]CaSR homodimers are the key regulators of calcium-sensing in FHH patients.[br][br]Nothing to Disclose: MPG, AS, GEB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1736 72 741 SAT-663 PO17-01 Saturday 680 2012


681 ENDO12L_SAT-664 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) The Efficacy of Adenovirus Delivery Systems To Investigate the Role of CREB and ICER in GnRH Regulation of LH Beta and FSH Beta Transcription in Primary Rat Pituitary Cells Daniel J Haisenleder, Kevin W Aylor, John C Marshall University of Virginia, Charlottesville, VA The intracellular pathways that mediate GnRH action on the gonadotrope remain to be fully characterized. cAMP regulates LH and FSH beta promoter activity, and GnRH increases cAMP and stimulates positive mediators of the cAMP/PKA pathway (i.e. CREB, CREM). Recently we showed that PACAP acts as an intermediary between GnRH actions and the cAMP pathway in gonadotropes, particularly on LH beta. GnRH also stimulates ICER, a competitive inhibitor of CREB activity, suggesting a potential role in the differential regulation of LH beta vs FSH beta transcription by pulsatile GnRH. To investigate these questions further, Sh-ICER and CREB dominant/negative (DN) constructs were cloned, packaged into adenovirus vectors, amplified and purified.[br]Pituitary cells from adult female rats were pretreated with Sh-ICER, CREB DN or control vector (viral dose = 10 MOI, multiplicity of infection) for 24h, then 48h later given 1nM GnRH (3 pretreatment pulses over 6 hours, followed by a final 10 min GnRH stimulus). LH beta, FSH beta and ICER primary transcripts (PTs) and ICER mRNA were measured by real time PCR. In control vector-treated cells, GnRH increased LH beta PT (2.1 fold), FSH beta PT (2 fold), ICER PT (1.5 fold) and ICER mRNA (1.7 fold) (p[lt]0.05 vs vehicle controls). As expected, Sh-ICER pretreatment suppressed basal and blocked GnRH-induced increases in ICER mRNA (p[lt]0.05), but had no effect on the ICER PT response to GnRH (1.6 fold increase; p[lt]0.05). Sh-ICER also reduced the stimulatory effect of GnRH on LH beta PT by 60% (p[lt]0.05). In contrast, Sh-ICER suppressed basal FSH beta PT, which enhanced the magnitude of the response to GnRH by 50% (p[lt]0.05). CREB DN pretreatment had no effect on basal LH beta PT, but completely suppressed the response to GnRH. In contrast, CREB DN reduced basal and GnRH-stimulated increases in FSH beta PT by 30%, but the magnitude of the response to GnRH was sustained.[br]In summary, suppressing expression of the endogenous inhibitor of the cAMP pathway, ICER, differentially regulates gonadotropin subunit genes by reducing the magnitude of LH beta and enhancing FSH beta responses to GnRH. In contrast, as previously seen following PACAP receptor blockade, inhibiting CREB activation selectively suppresses the stimulatory action of GnRH on LH beta transcription. Thus, adenovirus-delivered inhibitors of CREB and ICER are effect tools in studying mediators of the cAMP/PKA pathway within primary rat pituitary cells.[br][br]Nothing to Disclose: DJH, KWA, JCM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1107 73 742 SAT-664 PO19-02 Saturday 681 2012


682 ENDO12L_SAT-665 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Expression of the Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) Type 1 Receptor (PAC1R) Potentiates the Effects of GnRH on Gonadotropin Subunit Gene Expression Haruhiko Kanasaki, Aki Oride, Mijiddorj Tselmeg, Sukhbaatar Unurjargal, Kohji Miyazaki Shimane University School of Medicine, Izumo, Japan We examined the effect of the pituitary adenylate cyclase-activating polypeptide (PACAP) type 1 receptor (PAC1R) on gonadotropin-releasing hormone (GnRH)-induced gonadotropin subunit promoter activities using the L[beta]T2 gonadotroph cell line. In mock transfected cells, GnRH-increased LH[beta] and FSH[beta] promoters up to 2.74[plusmn]0.15 -fold and 1.6[plusmn]0.05 -fold respectively. When cells were transfected with PAC1R, both LH[beta] and FSH[beta] promoter activities were further increased up to 6.1[plusmn]0.87 -fold and 2.22[plusmn]0.43 -fold following GnRH stimulation. ERK phosphorylation, serum response element (SRE) promoters, and cAMP response element (CRE) promoters stimulated by GnRH were also potentiated in the presence of increasing amounts of PAC1R. The EC50 values for LH[beta] and FSH[beta] gene transcription by GnRH were significantly decreased by overexpression of PAC1R. PACAP 6-38, a PACAP receptor antagonist, failed to reduce the effect of GnRH on gonadotropin promoter activities in PAC1R overexpressing cells, suggesting that the potentiation of the effects of GnRH by PAC1R expression was not related to an autocrine mechanism of PACAP produced in the gonadotrophs. Our current results show that the action of GnRH in the regulation of gonadotropin subunit expression is enhanced by the presence of PAC1Rs.[br][br]Nothing to Disclose: HK, AO, MT, SU, KM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 862 73 743 SAT-665 PO19-02 Saturday 682 2012


683 ENDO12L_SAT-666 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Possible Involvement of PACAP and PACAP Type 1 Receptor in GnRH-Induced FSH [beta]-Subunit Gene Expression Aki Oride, Haruhiko Kanasaki, Indri Purwana, Tselmeg Mijiddorj, Sukhbaatar Unurjarga, Kohji Miyazaki Shimane University School of Medicine, Izumo, Japan Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) and its receptor, PACAP type 1 receptor (PAC1-R) play an important role in the induction of pituitary gonadotropins. In this present study, we examined whether the PAC1 -R was involved in the action of gonadotropin-releasing hormone (GnRH) on gonadotropin FSH[beta] subunit expression. In a static culture, GnRH stimulation significantly increased PAC1-R expression as well as PACAP gene expression in the gonadotroph cell line, L[beta]T2. Stimulation with low frequency GnRH pulses, which preferentially increase FSH[beta], increased the expression of both the PAC1-R and the PACAP genes to a greater extent than did high frequency pulses.[br]In the determination of transcriptional activity, the GnRH antagonist, cetrotide inhibited GnRH-induced FSH[beta] promoter activity completely, but PACAP6-38, a PACAP antagonist, had no effect on GnRH-induced FSH[beta] promoter activity. As expected, PACAP-induced FSH[beta] promoter activity was significantly prevented by PACAP6-38, but was not affected by cetrotide. PACAP6-38, however, significantly prevented GnRH-increased FSH[beta] mRNA expression. These observations suggest that GnRH-induced FSH[beta] gene expression is stimulated partially through PAC1-R by gonadotrophs producing PACAP or PAC1-R.[br][br]Nothing to Disclose: AO, HK, IP, TM, SU, KM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1562 73 744 SAT-666 PO19-02 Saturday 683 2012


684 ENDO12L_SAT-667 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Glucose Modulates Gonadotropin-Releasing Hormone (GnRH) Signaling in the Pituitary Gonadotrope Charukeshi Chandrasekera, Anita Mallya, Francina Gonzalez De Los Santos, Colleen Buggs-Saxton Wayne State University, Detroit, MI It is well known that women with poorly controlled diabetes have a spectrum of reproductive disorders such as delayed menarche, menstrual irregularity, and infertility. Although hyperglycemia alters hypothalamic and ovarian function, effects of hyperglycemia on the pituitary gonadotrope are presently unknown. Thus, we investigated whether hyperglycemia alters pituitary gonadotrope function.[br]Experiments were conducted in the mouse gonadotrope cell line (LT2). Cells were acclimated to media containing physiologic glucose concentration (100 mg/dL) and displayed no difference in viability compared to cells grown in standard media (450 mg/dL glucose). Hyperglycemia was induced by exposing LT2 cells to media containing elevated glucose concentration (600 mg/dL) for 24 hours followed by treatment with gonadotropin-releasing hormone (GnRH, 10 nM) for 2 hours. RNA was isolated and real-time PCR analysis was performed with Taqman gene expression assays for leutinizing hormone subunit (LH) and follicle stimulating hormone subunit (FSH). All data were normalized to an endogenous control gene and fold effects determined by comparison of GnRH-treated cells to control cells (glucose at 100 mg/dL with no GnRH) using the Ct method.[br]In LT2 cells grown in media with physiologic glucose (100 mg/dL), GnRH stimulated LH mRNA 1.65-fold and FSH mRNA 1.53-fold. When these cells were exposed to hyperglycemia (600 mg/dL glucose), the induction of FSH mRNA by GnRH was completely inhibited while LH mRNA induction by GnRH was not altered. Hyperglycemia also impaired PMA and forskolin-induced FSH activation, but did not alter activin-stimulated FSH mRNA. Metformin or insulin treatment did not reverse the inhibitory effect of hyperglycemia on GnRH-induced FSH mRNA. However, normalizing glucose concentrations back to physiologic concentration (100 mg/dL) completely reversed the inhibition of GnRH-stimulated FSH gene expression.[br]Taken together, our data indicate for the first time that hyperglycemia directly alters gonadotrope function by differentially modulating effects of GnRH on LH and FSH gene expression. It is well known that changes in serum levels of LH and FSH are associated with menstrual irregularity and infertility in diabetes. Thus, our observations suggest that hyperglycemia may cause dysregulation of the hypothalamic-pituitary-ovarian axis by modulating GnRH signaling pathways in the pituitary gonadotrope.[br][br]Sources of Research Support: Wayne State University School of Medicine. Children[apos]s Hospital of Michigan.[br][br]Nothing to Disclose: CC, AM, FGDLS, CB-S 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1609 73 745 SAT-667 PO19-02 Saturday 684 2012


685 ENDO12L_SAT-668 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Expression of GnRH, GnRH Receptor, Insulin Receptor and Androgen Receptor in a Rat Model of Polycystic Ovary Syndrome Gustavo A Maciel, Rodrigo R Marcondes, Katia C Carvalho, Gisele Giannocco, Vinicius C Amaral, Edmund C Baracat Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Universidade Federal de S[atilde]o Paulo [ndash] Escola Paulista de Medicina, S[atilde]o Paulo, Brazil CONTEXT: Polycystic ovary syndrome is a common endocrine condition, which is inked to hyperandrogenism and insulin resistance. In animals, androgen excess exposure in early life induces to abnormalities in female endocrinology in adulthood, such as permanent estrous. It is thought to be due a disruption of gonadotropin-releasing hormone (GnRH) secretion by the hypothalamus, but the exact mechanisms of it are uncertain. OBJECTIVE: To determine whether neonatal exposure to testosterone and estrogen alters the expression of GnRH, GnRH Receptor, Androgen Receptor and Insulin Receptor genes in the hypothalamus of female rats. METHODS: Fifteen female rats aged 0-3 days were sorted in three experimental groups according with subcutaneous administration of the following compounds: testosterone propionate (1,25 mg) (testosterone group, TG; n=5), estradiol benzoate (0,5 mg) (estradiol group, EG; n=5) and vehicle (0,1 mL) (control group, CG; n=5). With 90 days of age, the animals were sacrificed and the anterior portion of the hypothalamus was removed. Total RNA was extracted and cDNA synthesis was performed to evaluate the expression of genes GnRH, GnRHR, Insulin Receptor and Androgen Receptor using quantitative Real Time PCR. Statistical analysis was done using ANOVA and Tukey[apos]s multiple comparison tests. RESULTS: TG animals have a significant increase in AR expression (Relative expression [RQ] mean[plusmn]SD=1,176[plusmn]0,125) than EG (RQ=1,081[plusmn]0,033) and CG (RQ=1,000[plusmn]0,000) (p[lt]0,01). There was no change in relative expression [ndash] compared to the control group [ndash] of GnRH, GnRHR and IR genes (p=0.365, 0.052 and 0.112, respectively). Interestingly, expression of GnRHR gene was upregulated in Testosterone group (TG) and downregulated in Estradiol group (EG). There was a significant difference between TG and EG groups (p=0.018). All animals in the treated group (TG and EG) presented permanent estrous. CONCLUSION: Neonatal exposure to Testosterone or Estradiol alters the gene expression of Androgen Receptor but not of GnRH or Insulin receptor genes. Testosterone increases the expression of GnRH Receptor gene. Compensatory mechanisms and expression of other genes that regulate GnRH function might explain these findings.[br][br](1)Franks S. Medicine. 2009;37(9):441-44. (2)Dumesic DA et al. Reprod Fertil Develop. 2005;17:349-60. (3)Abbott DH et al. Reprod Biol Endocrinol. 2006;4:17. (4)Krsmanovic LZ et al. Trends Endocrinol Metab. 2009;20:402-8.[br][br]Sources of Research Support: Fundacao de Ampara a Pesquisa do Estado de Sao Paulo - FAPESP - 2010/17417-3.[br][br]Nothing to Disclose: GAM, RRM, KCC, GG, VCA, ECB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1243 73 746 SAT-668 PO19-02 Saturday 685 2012


686 ENDO12L_SAT-669 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Identification of a Bimodal Pattern of Preovulatory LH Surges in Female Mice Housed under a Bifurcated Circadian Paradigm Azim R Khan, Joshua Kim, Michael R Gorman, Alexander S Kauffman University of California, San Diego, La Jolla, CA; University of California, San Diego, La Jolla, CA; University of California, San Diego, La Jolla, CA Ovulation is triggered by a surge in pituitary luteinizing hormone (LH) secretion, which is itself induced by a surge in gonadotropin-releasing hormone (GnRH) release from the brain. In rodents, the timing of the GnRH/LH surge is controlled by a complex interaction between ovarian hormones and the circadian clock and occurs exclusively in the early evening, concurrent with the onset of locomotor activity. However, the mechanisms by which hormones and circadian rhythms interact to generate and time the GnRH/LH surge remain ill-defined. Recently, an experimental light cycle paradigm was developed for rodent models to explore alterations in circadian rhythmicity and entrainment. In lieu of a typical 24-h day alternating between subjective day and night in the light and dark phases, respectively, this paradigm exposes rodents to a [quot]bifurcated[quot] (split) 24-h lighting schedule comprised of two separate 5-h subjective nights (light:dark:light:dark; LDLD). Typically, rodents on such a LDLD schedule are active during only one of the two dark phases. However, when exposed to very dim illumination during the dark phases, mice and hamsters consistently show bimodal circadian behavior (active during both dark phases). While this bifurcated locomotor rhythm has been repeatedly confirmed, the circadian control of reproduction has not been examined in this context. Here, we studied timing of the circadian-based LH surge in bifurcated female mice. We hypothesized that if the two locomotor activity bouts in bifurcated mice reflect the output of two circadian oscillators or an accelerated single oscillator, then an LH surge may occur at the onset of both dark phases. To test this, we assessed LH surges in ovariectomized, estradiol-treated mice displaying stable bifurcated locomotor rhythms. We found that at least [sim] 63% of the bifurcated females displayed robust LH surges at the onset of one of the two subjective nights, while no bifurcated females (0%) had LH surges during the subjective day, suggesting that bifurcated mice may exhibit more than one LH surge per 24 h. We are currently determining if GnRH neuron activation occurs unilaterally in these bifurcated surging mice, as in other [quot]split[quot] paradigms. Our findings are the first demonstration of bifurcated circadian rhythms in female mice, extend our knowledge of the circadian flexibility induced by dim nighttime illumination, and offer further insight into the circadian mechanisms controlling reproduction.[br][br]Sources of Research Support: NSF IOS-1025893; NIH U54 HD012303 awarded to ASK.[br][br]Nothing to Disclose: ARK, JK, MRG, ASK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1062 73 747 SAT-669 PO19-02 Saturday 686 2012


687 ENDO12L_SAT-670 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Epigenetic Regulation during Gonadotrope Maturation Huimin Xie, Anitia K Iyer, Pamela L Mellon University of California, San Diego, La Jolla, CA Gene regulation is controlled, not only by a highly orchestrated cascade of transcription factors, but also by epigenetic mechanisms that include the modulation of chromatin structure and histone modifications. Gene activation and repression are specifically regulated through changes in chromatin structure imparted by chromatin remodeling, DNA acetylation, methylation, and other histone modifications. The developmentally staged gonadotrope cell lines, precursor [alpha]T1-1, immature [alpha]T3-1, and mature L[beta]T2, are useful cell models to address the epigenetic regulation status of gonadotrope-specific genes ([alpha]GSU, GnRHR, FSH[beta], and LH[beta]) during gonadotrope maturation. Three classes of chromatin structure have been described: active, poised, and repressed. We show, using DNaseI sensitivity assays, that [alpha]GSU stays in an active state in all three cell lines, while GnRHR is in an intermediate, poised state in [alpha]T1-1 cells, and becomes active in [alpha]T3-1 and L[beta]T2 cells. FSH[beta] and LH[beta] are active only in mature L[beta]T2 cells, consistent with specific expression patterns in these cell lines. We also performed ChIP assays to analyze histone modifications of all four genes during gonadotrope maturation assessing markers of active chromatin, Histone H3 acetylation (H3-Ac), H3K4 trimethylation (H3K4-Me3), RNA Polymerase II (RNAPII), and Phospho-RNAPII (p-RNAPII) binding, as well as the inactive chromatin marker, H3K9-Me2. The early gonadotrope-specific gene, [alpha]GSU, acquires open chromatin structure during gonadotrope maturation and GnRHR acquires, not only open chromatin structure, but also several histone modifications including H3-Ac, H3K4-Me3, and p-RNAPII. LH[beta] gene acquires a relatively open chromatin status in L[beta]T2, bearing H3-Ac and H3K4-Me3, though no significant epigenetic changes of FSH[beta] gene chromatin were detected as of yet.[br][br]Sources of Research Support: NIH R01 HD020377, U54 HD012303, R01 DK044838, T32 DK007044, T32 HD007203, F32 HD070579, F32 HD058427, T32 DK007494.[br][br]Nothing to Disclose: HX, AKI, PLM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 971 73 748 SAT-670 PO19-02 Saturday 687 2012


688 ENDO12L_SAT-671 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Epigenetic Control of GnRH Neuron Development: Role of Histone Deacetylase 9 (HDAC9) Smita Salian-Mehta, Mei Xu, Emily Busta, Todd Horn, Timothy McKinsey, Margaret E Wierman University of Colorado Denver, Aurora, CO; Veterans Affairs Medical Center Denver, Denver, CO; University of Colorado Denver, Aurora, CO Disruption of Gonadotropin releasing hormone (GnRH) neuron function results in abnormal sexual maturation and infertility in mice and humans. However, epigenetic modulation of GnRH neuron development is largely unexplored. DNA microarrays revealed that the undifferentiated NLT GnRH neuronal cells expressed uniformly lower histone deacetylases (HDACs) and DNA methyl transferases (DNMTs) transcripts as compared to differentiated GT1-7 cells. Although initially described as global transcriptional repressors, HDAC subfamily members have other cell specific roles. HDAC activity was assessed using acetylated class specific HDAC substrates that once deacetylated by endogenous HDACs are susceptible to cleavage by trypsin and release a fluorophore. Class I HDAC (HDACs1/2) and Class IIb HDAC (HDAC 6) activity in GT1-7 cells was increased 1.7 fold and 5.7 fold respectively as compared to NLT. The Class IIa HDAC9 that interacts with MEF2, a transcription factor earlier implicated in GnRH gene expression and neuronal survival, was increased (6.7 fold) in GT1-7 compared to NLT GnRH neurons. Differential HDAC9 expression was confirmed at the mRNA level by RT-PCR (2.9 fold) and protein level by immunoblot (10 fold). Although overall Class IIa HDACs (4/5/7/9) activity levels were similar, immunoprecipitation of neuronal lysates with HDAC9 antibody confirmed increased HDAC9 activity in GT1-7 cells as compared to NLT (1.5 fold). Over-expression or silencing of HDAC9 in GT1-7 cells had no effect on endogenous GnRH gene expression Gn cells as assessed by RT-PCR. However, HDAC inhibitors, TSA (blocks Class I/II) and DPAH (blocks Class IIa) increased cleaved caspase 3 as an index of apoptosis (2.7 fold and 1.5 fold, respectively) suggesting a role of Class IIa HDACs in survival of GnRH neurons. Over-expression of HDAC9 mRNA (9 fold) and protein (23 fold) in NLT GnRH neuronal cells (low endogenous HDAC9) decreased caspase-3 cleavage compared to control (0.8 vs.2.3 fold). Alternatively, silencing of HDAC9 in GT1-7 cells (90%) selectively increased caspase-3 cleavage (1.8 vs. 2.4 fold compared to control) confirming the pro-survival role of HDAC9. In migration assays, over-expression of HDAC9 blocked NLT neuron migration (43%), suggesting that HDAC9 also acts as a stop signal for GnRH neuron migration. Together these data support the epigenetic effects of HDAC9 to promote neuron survival and inhibit GnRH neuron movement across neuronal development.[br][br]Sources of Research Support: HD32119 to MEW.[br][br]Nothing to Disclose: SS-M, MX, EB, TH, TM, MEW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2076 73 749 SAT-671 PO19-02 Saturday 688 2012


689 ENDO12L_SAT-672 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Sex Steroid and Leptin Receptors in Gonadotropin Inhibitory Hormone Neurons Iain J Clarke, Alda Pereira, Qun Li, Jeremy T Smith Monash University, Clayton, Australia GnIH-3 is a peptide produced in neurons in the dorsomedial nucleus/paraventricular nucleus in the sheep, as in other mammals (1). GnIH-3 is encoded by the GnIH gene that also encodes GnIH-1. Most work in mammals has been done with GnIH-3, demonstrating inhibitory effects on the gonadotropin releasing hormone (GnRH) [ndash] gonadotropin axis. In sheep, the neurons project to the median eminence and the GnIH is secreted into the hypophysial portal blood (2), to act on the pituitary gonadotropes. We have shown that GnIH gene expression is reduced in the follicular phase of the estrous cycle, but infusion of GnIH-3 blocks the estrogen-induced preovulatory-like LH surge (3). This suggests that lowering of GnIH tone is permissive of estrogen action to induce the preovulatory LH surge. This raises the question as to whether the GnIH neurons express sex steroid receptors, allowing direct feedback effect. In addition to having a negative effect on the reproductive axis, GnIH also stimulates food intake (3). Accordingly, a further question is whether these neurons respond to feedback hormones, such as leptin, that regulate brain systems maintaining metabolic homeostasis. The aim of this study was to determine the level of expression of estrogen, progesterone and leptin receptors in ovine GnIH neurons. Brains of ewes (n=4) were perfusion fixed with paraformaldehyde and processed for either in situ hybridisation or immunohistochemistry. Co-localisation of estrogen receptor (ER)-[alpha] and ER-[beta], progesterone receptor (PR) to GnIH-3 neurons was examined by double-labelling immunofluorescence histochemistry. Leptin receptor (ObRb) co-localisation was determined by in situ hybridisation ([sup]35[/sup]S-labelling for receptor and Digoxigenin for GnIH). The level of expression of ER-[alpha] and PR was low (20% and 27% respectively), whereas 59% of GnIH neurons expressed ER-[beta] and half (54%) expressed ObRb. These data suggest that regulation of the GnIH neurons through the main sex steroid receptors involved in feedback effects on the GnRH/gonadotropin axis (ER-[alpha] and PR) is minimal, whereas major regulation of the neurons via ER-[beta] seems likely. Indirect regulation through other sex-steroid responsive is possible. The level of ObRb expression suggests that at least half of the GnIH neurons are responsive to metabolic state.[br][br]1. Clarke et al (2008) Endocrinology 149: 5811. 2. Smith et al. (2011) Proc Endoc Soc Abstract P1-375. 3. Clarke et al (2012) Neuroendocrinology, EPub 25 Jan PMID: 22286004.[br][br]Nothing to Disclose: IJC, AP, QL, JTS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 850 73 750 SAT-672 PO19-02 Saturday 689 2012


690 ENDO12L_SAT-673 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Influence of Sex Steroids on the Expression of RFRP-3 (GnIH) and Co-Expression of Its Receptor, Gpr147, in Neuropeptide Populations in Adult Mice Matthew C Poling, Alexander S Kauffman University of California San Diego, La Jolla, CA; University of California San Diego, La Jolla, CA; University of California San Diego, La Jolla, CA RFamide-related peptide 3 (RFRP-3; mammalian ortholog to Gonadotropin-inhibitory hormone [GnIH]) is a recently-identified neuropeptide that can inhibit LH release and GnRH neuronal activity. In addition to its reproductive function, RFRP-3 can influence food intake, body temperature, and stress responses. It has been proposed that RFRP-3 may participate in sex steroid-mediated feedback control of GnRH neurons, but data published thus far in rodents is equivocal. Using in-situ hybridization, we assessed whether the expression of [italic]Rfrp[/italic] (the gene encoding RFRP-3) changes in adult male and female mice that were gonadectomized (GDX) and treated with estradiol (GDX+E), testosterone (GDX+T), dihydrotestosterone (GDX+DHT), or nothing. We found that E and T significantly repressed [italic]Rfrp[/italic] expression. In contrast, DHT had no effect on [italic]Rfrp[/italic] expression. Next, we performed double label in-situ hybridization to determine if estrogen receptor [alpha] ([italic]ER[alpha][/italic]) or androgen receptor ([italic]AR[/italic]) is expressed in [italic]Rfrp[/italic] neurons in male and female mice. [italic]ER[alpha][/italic] was co-expressed in [sim]25% of [italic]Rfrp[/italic] neurons while practically none of the [italic]Rfrp[/italic] neurons express [italic]AR[/italic]. These findings suggest that sex steroids may directly repress [italic]Rfrp[/italic] expression in a subset of RFRP-3 neurons via an ER (but not AR) pathway. Moreover, the inhibition of [italic]Rfrp[/italic] by sex steroids supports a role of RFRP-3 in positive, and not negative, feedback control of GnRH. To assess whether RFRP-3 might regulate the reproductive axis by acting directly on GnRH neurons, we used double label in-situ hybridization to quantify the expression of RFPR-3[apos]s receptor, Gpr147, in GnRH neurons. [italic]Gpr147[/italic] was expressed in a small subset of GnRH neurons ([sim]15%) in both male and female mice, and this co-expression was unaffected by the sex steroid milieu. Thus, RFRP-3 might act directly on some GnRH neurons, though some of RFRP-3 effects on GnRH could also be mediated indirectly. To test this possibility, we examined the co-expression of [italic]Gpr147[/italic] in kisspeptin ([italic]Kiss1[/italic]) neurons in the anteroventral periventricular nuclear continuum (AVPV/PeN) of female mice. Only a small proportion of [italic]Kiss1[/italic] neurons co-expressed [italic]Gpr147[/italic] ([sim]15%) in diestrous females, and the degree of co-expression was lower ([lt]9%) in GDX+E treated mice. Current experiments are examining the co-expression of [italic]Gpr147[/italic] in other neuropeptide populations in the hypothalamus in order to better understand RFRP-3[apos]s role in reproductive endocrinology and non-reproductive processes.[br][br]Sources of Research Support: National Science Foundation grant IOS-1025893; NIH grant R01 HD065856; Diabetes and Endocrinology Research Center (DERC) Pilot and Feasibility grant (P30 DK063491); Eunice Kennedy Shriver NICHD/NIH (SCCPIR) grant U54-HD012303.[br][br]Nothing to Disclose: MCP, ASK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 274 73 751 SAT-673 PO19-02 Saturday 690 2012


691 ENDO12L_SAT-674 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Central Mechanisms for the Direct Inhibitory Effects of Gonadotropin-Inhibitory Hormone (GnIH) on Gonadotropin-Releasing Hormone (GnRH) Using Novel Hypothalamic Cell Models Nicole Gojska, Denise D Belsham University of Toronto, Toronto, Canada; Toronto General Hospital Research Institute, University Health Network, Toronto, Canada Reproduction is coordinated by the actions of specific neuropeptides and peripheral hormones, all of which converge on the gonadotropin-releasing hormone (GnRH) neurons, which reside at the pinnacle of the hypothalamic-pituitary-gonadal (HPG) axis. Recently, a novel hypothalamic neuropeptide, gonadotropin-inhibitory hormone (GnIH), has emerged as a potent inhibitory modulator of neuroendocrine function. In mammals, GnIH (also referred to as RFRP-3) appears to be localized in the dorsomedial hypothalamus and displays similar inhibitory functions as the avian form, though its distinct role in the HPG axis is not well established. To date, there is a paucity of studies focusing on the regulation of hypothalamic GnIH, as well as its potential direct regulation of GnRH neurons. We have generated immortalized, clonal, rodent cell lines derived from both embryonic and adult hypothalamic primary culture. Using semi-quantitative RT-PCR, we have classified a subset of cell lines that exhibit strong GnIH expression, as well as receptors for glucocorticoids (GR) and estrogen (ER[beta] and GPR30). We have also verified the presence of mammalian GnIH in two cell lines, rHypoE-19 and rHypoE-23, using an anti-RFRP antibody (PAC 1365; generously provided by Dr. L.J. Kriegsfeld, UC Berkeley). In a newly established cell model of GnRH neurons, the mHypoA-GnRH/GFP, we confirmed the presence of the GnIH receptor, GPR147. These neurons were immortalized and FAC-Sorted from an adult GnRH-GFP mouse to generate a non-clonal cell line representative of the entire GnRH neuronal population. Using real time RT-PCR we analyzed the dose-dependent and direct effects of GnIH on GnRH mRNA expression. We have demonstrated that GnIH treatment (100 nM) directly represses GnRH mRNA expression by approximately 40% at 1 and 4 hours (P[lt]0.05). Current studies using the GPR147 antagonist, RF9, and Western blot analysis are being used to delineate the direct GnIH/GPR147-mediated mechanisms controlling GnRH transcription. Preliminary evidence suggests that RF9 can attenuate the inhibitory actions of GnIH on GnRH expression. Future studies will explore the effects of GnIH on GnRH secretion, through both direct GnIH treatments and co-culture experiments. We anticipate that these novel hypothalamic GnIH cell models can be used to further define the cellular mechanisms by which GnIH input and signaling mediates mammalian reproduction through the modulation of GnRH.[br][br]Sources of Research Support: This research was supported by CIHR, CFI, CRC, and OGS.[br][br]Nothing to Disclose: NG, DDB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1700 73 752 SAT-674 PO19-02 Saturday 691 2012


692 ENDO12L_SAT-675 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) A Novel C-Terminal Heptapeptide Contributes to the Regulated Secretion of LH Initiated at Unique Sub-Domains of the ER Albina Jablonka-Shariff, Irving Boime Washington University School of Medicine, St Louis, MO The coordinated secretion and activity of pituitary lutropin (LH) and follitropin (FSH) are critical for gonadal reproductive function. LH is released through the regulated pathway, i.e., it is released by secretagogues, while FSH is secreted constitutively. Although both unassembled gonadotropin [beta] subunits accumulate in the ER and are poorly secreted, co-expression with the common [alpha] subunit rescued them quantitatively except for LH[beta] subunit. Previously, we identified a novel C-terminal heptapeptide sequence detected in only the LH[beta] subunit (115-Leu-Ser-Gly-Leu-Leu-Phe-Leu-121), as a sorting signal that mediates entry of the LH dimer into the regulated secretory pathway in transfected rat somatotrope-derived GH[sub]3[/sub] cells, where both the constitutive and regulated secretory pathways operate. We previously identified another feature of the heptapeptide, its ability to direct the LH[beta] subunit to a perinuclear sub-domain of the ER, which is distinct from localization of the FSH[beta] subunit. Similar to LH[beta], an FSH[beta] chimera containing the heptapeptide displayed similar perinuclear-staining pattern. Moreover, both the LH[beta] and FSH[beta] chimera were co-localized with the ER luminal chaperone BiP, but not membrane-bound calnexin. In contrast, a mutant LH[beta] devoid of the heptapeptide and the wild-type FSH[beta] and [alpha] subunits displayed dispersed cytoplasmic puncta without detectable perinuclear staining. These data show that LH[beta] and FSH[beta] subunits were detected in distinct regions of the ER, and the LH[beta] heptapeptide was critical for this ER sorting. Here, we tested this hypothesis by examining LH[beta] and FSH[beta] immunolocalization in transfected PC12 cells, which like GH[sub]3[/sub] cells contain a regulated secretion pathway. A similar perinuclear localization of LH[beta] was observed, whereas, FSH[beta] was detected only in the peripheral ER. Because CHO and MDCK cells lack regulated secretory pathways, we also examined the fluorescence staining pattern of the LH[beta] subunit in these cells. In contrast to GH[sub]3[/sub] and PC12 cells, both cell lines expressing LH[beta] showed only dispersed cytoplasmic puncta, indicative of peripheral ER localization. These results support the hypothesis that the perinuclear accumulation of secretory proteins is associated with the regulated secretion. A model is proposed in which the C-terminal heptapeptide sequence mediates heterodimer assembly within a sub-domain of the ER/nuclear envelope as an early partitioning event for the entrance of LH into the regulated secretory pathway.[br][br]Nothing to Disclose: AJ-S, IB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 382 73 753 SAT-675 PO19-02 Saturday 692 2012


693 ENDO12L_SAT-676 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Developmental Expression of GnRH-(1-5) Receptor in a Rat Model Eleane Beadle, D Omar Larco, Adam Vana, Madelaine Clark, T John Wu National Capital Consortium, Bethesda, MD; Uniformed Services University of the Health Sciences, Bethesda, MD Purpose of Study: Gonadotropin releasing hormone (GnRH) is a hormone that plays an essential role in control of reproductive function and behavior. Previous studies have demonstrated that GnRH-(1-5), an active metabolite of GnRH, has a different effect on hormone levels and reflex reproductive behavior when compared to its parent peptide. At present, the biologic receptor for GnRH-(1-5) is unknown, but a candidate receptor, the G-protein coupled receptor 101 (GPR101), has been identified based on a high-throughput affinity binding assay. In the present study, levels of hypothalamic GPR101 mRNA expression were compared between the pre-pubertal and post-pubertal rat.[br]Methods: Sprague-Dawley rats were euthanized prepubertally (P21) and postpubertally (P60). Brains were collected and samples were processed to synthesize cDNA. Quantitative PCR was used to determine the expression of GPR101 mRNA. RNA integrity was assessed by gel electrophoresis and the PCR product specificity was confirmed by sequencing. Primary outcome for GPR101 was normalized to beta-actin. Statistical significance was set at p[lt]0.05.[br]Summary of Results: GPR101 mRNA expression was detected in both adults and pups at levels consistent with other biologically active receptor genes. Expression of GPR101 mRNA was higher in the hypothalamus of adult females when compared to pup females (n=12, p[lt]0.05). Greater hypothalamic expression of GPR101 mRNA in adults compared to pups was also seen in males (n=12, p[lt]0.05). For all ages and sex, the hypothalamic expression of GPR101 mRNA was significantly higher than in the cortex (p[lt]0.05). Unlike the hypothalamus, there was no age or sex difference (p[gt]0.05) of GPR101 mRNA in the cortex.[br]Conclusion: Increased levels of GPR101 mRNA expression in the hypothalamus after puberty support the hypothesized role of GPR101 in reproductive function. Further studies are needed to demonstrate GPR101 as the biologically active receptor for GnRH-(1-5), but the results of the current studies suggest that it may be important in reproduction.[br][br]Sources of Research Support: NSF Grant IOS-1052288 and DoD RO85FN.[br][br]Nothing to Disclose: EB, DOL, AV, MC, TJW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1816 73 754 SAT-676 PO19-02 Saturday 693 2012


694 ENDO12L_SAT-677 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Implication of the Wnt5a in the Differentiation and Maturation of GnRH-Secreting GnV3 Neurons Celine Campagne, Fabrice Marcillac, Sarah Geller, Virginie Mansuy, Francois P Pralong, Vincent E Prevot University Hospital, Lausanne, Switzerland; Inserm U837, Lille, France; INRA UMR 6175 CNRS, Nouzilly, France Hypothalamic GnRH neurons are the main regulators of reproduction. Despite a widespread and long-lasting interest in the study of their ontogeny, the cellular and molecular processes implicated in their differentiation and maturation remain poorly understood. Here we used GnV-3 cells, a model of conditionnally immortalized GnRH-expressing neurons, to gain a better insight into the molecular mechanisms implicated in GnRH neurons development.[br]We first performed a transcriptomic analysis of GnV-3 cells at different stages of differentiation (1). These microarray data allowed us to identify the [italic] wnt/frizzled pathway [/italic] as potentially important for the development of GnV3 cells. These data were then confirmed by qRT PCR and western blot analyses. We thus found that GnV3 cells express Wnt5a, and that its expression is significantly increased in differentiated cells compared to proliferating ones. We next examined the pattern of expression of various receptors implicated in Wnt5a signalling, and found that Frizzled1, Frizzled2, Ryk and ROR2 are expressed by GnV3 cells. The use of phosphospecific antibodies allowed us to demonstrate activation of the [beta]-catenin pathway during GnV-3 cells differentiation.[br]A morphological analysis revealed that upon Wnt5a treatment (300ng/mL, 72h), the percentage of GnV3 cells adopting a bipolar phenotype is significantly increased in the proliferation state. However Wnt5a did not affect [beta]-catenin phosphorylation, suggesting an implication of another pathway.[br]Taken together, these data demonstrate that Wnt5a is promoting the initial stages of maturation of GnRH secreting GnV3 cells. Furthermore, the expression of Wnt5a by these cells is suggestive of an autocrine regulation. Overall, GnV3 cells appear as a valuable model for the [italic] in vitro [/italic] study of the molecular events underlying GnRH neurons development, and further work will be necessary to better investigate the cellular and molecular pathways involved in the effects of Wnt5a.[br][br](1) Mansuy et al., Mol Cell Endocrinol. 2011 Jan 30;332(1-2):97-105.[br][br]Nothing to Disclose: CC, FM, SG, VM, FPP, VEP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1219 73 755 SAT-677 PO19-02 Saturday 694 2012


695 ENDO12L_SAT-678 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) FOXO1 Diminishes Activin Signaling in Pituitary Gonadotrope Cells Rishi V Parikh, Chung Hyun Park, Varykina G Thackray University of California, San Diego, La Jolla, CA The production of FSH and LH from gonadotrope cells in the anterior pituitary is critical for reproduction. Synthesis of these hormones is tightly controlled by a complex network of hormonal signaling pathways including GnRH, activin and steroid hormones. There is also evidence that gonadotropin production is regulated by metabolic cues such as insulin. One group of candidate genes that may be regulated by insulin in gonadotropes is the FOXO subfamily of forkhead transcription factors. We have shown previously that the FOXO1 transcription factor inhibits basal and GnRH induction of Lhb synthesis. In the present study, we investigate whether FOXO1 modulates activin signaling in gonadotropes and explore mechanisms of action. Our studies show that overexpression of FOXO1 represses activin induction of Fshb, Lhb and Gnrhr gene expression in L[beta]T2 cells. In regards to the Fshb promoter, we demonstrate that mutation of the FOXO1 DNA-binding domain (DBD) relieves the suppression, indicating that the effect of FOXO1 requires direct DNA binding or protein-protein interactions via the DBD. In addition, FOXO1 suppression of activin induction maps to the 304/-95 region of the Fshb promoter containing multiple activin responsive regions. To determine whether FOXO1 suppression involves SMAD proteins, we tested whether FOXO1 repressed SMAD induction of the Fshb promoter. Interestingly, SMAD3/4 induction of Fshb was completely suppressed by FOXO1, suggesting that FOXO1 suppression of activin induction is due to interference with SMADs. We also show that the suppressive effects of FOXO1 map to the proximal region of the Gnrhr promoter. Similarly to Fshb, mutation of the FOXO1 DBD relieves the suppression of Gnrhr gene expression by FOXO1. Additional experiments are underway to further define the cis regulatory regions on the Fshb and Gnrhr promoters necessary for FOXO1 suppression and to explore how interactions among FOXO1 and SMADs contribute to the suppression. In summary, we show that FOXO1 diminishes activin induction of Fshb, Lhb and Gnrhr gene expression. These data also suggest that FOXO1 may play an important role in controlling gonadotropin levels in response to metabolic cues.[br][br]Sources of Research Support: NIH Grants R01 HD067448, K01 DK080467, P30 DK063491 (P[amp]F Award), U54 HD012303 and a UCSD Academic Senate Health Sciences Grant.[br][br]Nothing to Disclose: RVP, CHP, VGT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 236 73 756 SAT-678 PO19-02 Saturday 695 2012


696 ENDO12L_SAT-679 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Impaired Fertility and FSH Synthesis in Gonadotrope-Specific Foxl2 Knockout Mice Stella Tran, Xiang Zhou, Christine Lafleur, Sarah Kimmins, Derek Boerboom, Ulrich Boehm, Mathias Treier, Daniel J Bernard McGill University, Montr[eacute]al, Canada; McGill University, Ste-Anne de Bellevue, Canada; University of Montr[eacute]al, Ste-Hyacinthe, Canada; The Center for Molecular Neurobiology Hamburg, Hamburg, Germany; Max-Delbr[uuml]ck Center for Molecular Medicine, Berlin, Germany Impairments in follicle-stimulating hormone (FSH) synthesis or action have profound effects on fertility, particularly in females. Mechanisms controlling FSH synthesis by pituitary gonadotrope cells are incompletely understood. Previously, we discovered a novel and critical role for forkhead protein L2 (FOXL2) in activin-stimulated FSH[beta] subunit (Fshb) transcription in the immortalized murine gonadotrope-like cell line L[beta]T2. FOXL2 was similarly implicated in regulation of GnRH receptor (Gnrhr), gonadotropin [alpha] subunit (Cga), and follistatin (Fst) expression. To examine FOXL2[apos]s role in pituitary in vivo, we generated a gonadotrope-specific Foxl2 knockout model (hereafter cKO) by crossing [apos]floxed[apos] Foxl2 and GnRH receptor IRES Cre (GRIC) mice. cKO mice developed overtly normally but their ovarian and testicular weights were significantly reduced relative to controls in adulthood. Estrous cyclicity and fertility were impaired in cKO females, with the frequency and size of litters reduced by approximately 50%. Ovaries of cKO females expressed wild-type levels of FOXL2 and responded normally to exogenous gonadotropins, ruling out an ovarian defect. In contrast, the number of ovulated oocytes observed after natural mating was reduced in cKO relative to controls females. cKO males exhibited abnormalities in testicular morphology, had significantly reduced sperm counts, and sired slightly smaller litters than controls. Fshb, Fst, and Foxl2 mRNA levels were significantly reduced in pituitaries of adult cKO mice of both sexes relative to controls whereas Gnrhr, Lhb, and Cga mRNAs were unaffected. Serum FSH was dramatically reduced in cKO males and a similar trend was observed in randomly cycling females. To determine whether the observed FSH deficiency might derive from impaired activin signaling, we transduced primary pituitary cultures from floxed Foxl2 mice with Cre recombinase-expressing adenoviruses. Recombination of the Foxl2 alleles attenuated both basal and activin A-induced Fshb mRNA levels and FSH secretion without affecting luteinizing hormone release. Collectively, these data provide the first definitive demonstration of a gonadotrope-restricted transcription factor required for the selective regulation of FSH synthesis and secretion in vivo. In the absence of FOXL2, activin-stimulated Fshb expression and FSH secretion are reduced, leading to impairments in follicle development, spermatogenesis, and fertility.[br][br]Sources of Research Support: This research was supported by operating funds from the Canadian Institutes of Health Research (MOP 89991) to DJB and a research grant (BO1743/2) by the Deutsche Forschungsgemeinschaft (DFG) to UB. DJB and ST hold a Chercheur-boursier (Senior) and a doctoral fellowship, respectively, from the Fonds de la recherche en sant[eacute] du Qu[eacute]bec.[br][br]Nothing to Disclose: ST, XZ, CL, SK, DB, UB, MT, DJB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2127 73 757 SAT-679 PO19-02 Saturday 696 2012


697 ENDO12L_SAT-680 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Differential Expression of NELF Isoforms 2 and 3 in the Nucleus and Cytoplasm of GnRH Neuronal Cells Samuel D Quaynor, Lynn P Chorich, Richard S Cameron, Lawrence C Layman Georgia Health Sciences University, Augusta, GA Nasal embryonic luteinizing hormone releasing factor (NELF) was first isolated from migratory gonadotropin releasing hormone (GnRH) neurons in mice. Subsequently, the human ortholog was cloned and mutations were found in patients with pubertal disorders idiopathic hypogonadotropic hypogonadism (IHH) and Kallmann Syndrome (KS). We previously showed that NELF protein was more highly expressed in migratory GN11 and NLT cells compared with postmigratory GT1-7 immortalized GnRH neuronal cells. NELF was largely confined to the nucleus in NLT cells, and mutation of a putative nuclear localization signal of NELF rendered its expression diffusely throughout the cell. In the mouse, rat, and human, most NELF protein was intranuclear, although some protein expression was observed outside of the nucleus. Findings from western blot analysis indicated that at least several bands were present, which were probably due to multiple isoforms. We hypothesized that the different NELF isoforms will contribute to differential cellular expression patterns and account for additional bands on western blot. We have characterized the different NELF isoforms in immortalized mouse GnRH neurons by RT-PCR and sought to clone full length isoforms. RNA from NLT cells was used for RT-PCR to amplify the full-length isoforms, which was confirmed by DNA sequencing. The full-length cDNA was then cloned into pcDNA3.1 and transfected by electroporation into NLT cells which were then processed for western blot analysis using a c-myc antibody. Isoform 2, the most frequent isoform encountered in NLT cells was shown by immunofluorescence and western blot analysis to be localized to the nucleus and 64 kD. In contrast, isoform 3, which lacks the nuclear localization signal, is expressed throughout the cell and is 61 kD. These findings indicate that these two isoforms are translated into protein and explain the discordant expression patterns seen in GnRH neurons and the different sized bands by western blot analysis. These full-length isoforms now permit the identification of isoform-specific binding proteins, which is anticipated to shed light into the pathways in which NELF operates in the nucleus and cytoplasm.[br][br]Sources of Research Support: LL supported by HD33004 from the NICHD.[br][br]Nothing to Disclose: SDQ, LPC, RSC, LCL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1151 73 758 SAT-680 PO19-02 Saturday 697 2012


698 ENDO12L_SAT-681 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Proteasome Activity Is Required for Cyclic GnRH-Stimulated Exchange of Repressor and Activator Proteins from the LH[beta] Promoter Debalina Bagchi, Josefa Andrade, Margaret A Shupnik University of Virginia School of Medicine, Charlottesville, VA; University of Virginia School of Medicine, Charlottesville, VA Hypothalamic GnRH pulses with defined frequencies play a crucial role in modulating gonadotropin hormone secretion, and transcription of the subunit genes, a Luteinizing Hormone (LH[beta]) and Follicle Stimulating Hormone (FSH[beta]) throughout the reproductive cycle. LH[beta] is the most dramatically regulated by GnRH. Incorrect ratios of LH/FSH can lead to infertility syndromes like Polycystic ovarian syndrome. We examined GnRH-regulated transcription by chromatin immunoprecipiation (ChIP) assays to assess binding of transcription factors, co-activators and co-repressors to the LH[beta] promoter. Previously, we reported that with GnRH, associations of the orphan nuclear receptor SF1 and early growth response1 (Egr-1) protein were stimulated, coincident, and cyclical with a period of 30 min. Activated pRNA polymerase II also bound with a similar pattern. This cycling and recruitment of SF1,Egr-1 and pRNA POLII was prevented by the proteasome inhibitor MG132, suggesting an essential role of proteasomal degradation in recruitment of proteins to LH[beta] and hence its transcription. We hypothesized that removal of suppressor proteins by the proteasome may be required for stimulated rhythmic associations of SF1 and Egr-1. Co-repressor proteins could be associated with SF1, or suppressors could be directly bound with DNA. Our studies show that GnRH stimulates association of coactivators SRC-1 and GCN5 with the LH[beta] promoter, and MG132 completely prevents the GnRH-stimulated association. The co-repressor SMRT also associates with the promoter, but MG132 treatment maintains SMRT association. One candidate for a DNA binding repressor protein is WT1 (Wilms Tumor1), which binds to similar DNA motifs as Egr-1. We found WT1 is expressed in L[beta]T2 cells and binds the LH[beta] promoter in ChIP assays, suggesting it can modulate LH[beta] gene transcription. Cellular WT1 levels increase with MG132 treatment, and WT1 binding to the LH[beta] was also increased, correlating with decreased Egr-1 binding. Overall, our data suggest that proteasome activity is required to remove repressor proteins associated with transcription factors or directly binding on the LH[beta] promoter, and that this process is crucial for subsequent recruitment of coactivators and transcription factors, as well as GnRH-stimulated transcription of LH[beta].[br][br]Sources of Research Support: This work was supported by the Eunice Kennedy Shriver National Institute of Child Health andHuman Development/National Institutes of Health throughthrough cooperative agreement (U54 HD28934 project to MAS) as part of the Specialized Cooperative Centers Program in Reproduction and Infertility Research.[br][br]Nothing to Disclose: DB, JA, MAS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 282 73 759 SAT-681 PO19-02 Saturday 698 2012


699 ENDO12L_SAT-682 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) GnRH Signaling Via Reactive Oxygen Species Taeshin Kim, Mark A Lawson University of California, San Diego, La Jolla, CA Reactive oxygen species (ROS) are produced as a byproduct of the normal metabolism of oxygen and stimulate the dual specificity protein phosphatase 1 (DUSP1) and sulfiredoxin (SRX) to regulate cell signaling and homeostasis. Expression of the [italic]Dusp1[/italic] gene is induced by oxidative or heat stress and growth factors. We have shown that gonadotropin-releasing hormone (GnRH) also increases DUSP1 and that GnRH activation of [italic]Lhb[/italic] gene expression is suppressed by DUSP1. Sulforedoxin plays an important role in antioxidant metabolism by re-activating peroxiredoxins, a family of peroxidases, when these enzymes are inhibited by excess oxidation. In this study, we investigated ROS production and function during the signaling response to GnRH stimulation in LbetaT2 pituitary gonadotropes. GnRH induced ROS generation in LbT2 cells was observed by cell imaging after exposure to 2[prime]-7[prime] dichlorofluorescein diacetate. ROS generation was reduced by addition of N-acetyl-L-cysteine (NAC) as an antioxidant. Activation of ERK by GnRH was reduced by the presence of NAC, indicating ROS involvement in ERK activation by GnRH. Moreover GnRH-induced expression of [italic]Dusp1[/italic] mRNA was significantly reduced by NAC treatment. Furthermore the expression of [italic]Srx[/italic] mRNA was also increased by GnRH treatment in LbT2 cells, and was significantly reduced by addition of NAC. Control H[sub]2[/sub]O[sub]2[/sub] treatment significantly stimulated the expression of [italic]Dusp1[/italic] mRNA in LbT2 cells, which had an additive effect in addition to GnRH treatment. This observation suggests that excess oxidative stress may inhibit gonadotropin expression via increased DUSP1. These findings establish that GnRH causes ROS production in LbT2 gonadotropes, which regulates downstream signaling via ERK activation, [italic]Dusp1[/italic] and [italic]Srx[/italic] expression. This demonstrates the importance of ROS as physiological signaling molecules in the regulation of gonadotropins.[br][br]Sources of Research Support: NIH/NICHD Grant R01-HD043758.[br][br]Nothing to Disclose: TK, MAL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2030 73 760 SAT-682 PO19-02 Saturday 699 2012


700 ENDO12L_SAT-683 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) FOXO1 Inhibition of Follicle-Stimulating Hormone Beta Gene Transcription Danalea Skarra, David J Arriola, Varykina G Thackray University of California, San Diego, La Jolla, CA The gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), are necessary for mammalian fertility and are produced by gonadotrope cells located within the anterior pituitary. Stimulation of gonadotropes with insulin or growth factors modulates gonadotropin production, although the mechanisms are unknown. One candidate that may transduce insulin signaling into functional changes in gonadotropin synthesis is the Forkhead box-O (FOXO) family of transcription factors, which function as mediators of diverse cellular pathways. In response to insulin, FOXOs are negatively regulated by Akt phosphorylation, which results in their export from the nucleus to the cytoplasm, thereby limiting their nuclear activity. Our laboratory has shown that FOXO1 is expressed in murine primary pituitary gonadotropes and in LbT2 cells, an immortalized gonadotrope cell line. We have demonstrated that gonadotrope FOXO1 is phosphorylated by Akt on serine 256 in response to insulin, leading to FOXO1 cytosolic accumulation. Since we have previously shown that FOXO1 inhibits basal and GnRH-induced[italic] Lhb[/italic] transcription, we investigated the effect of FOXO1 on the synthesis of the beta subunit of FSH ([italic]Fshb[/italic]) in the current study. [italic]Fshb[/italic] synthesis is the rate-limiting step in FSH production and is closely coupled to FSH secretion, as it is secreted via the constitutive secretory pathway. We have found that FOXO1 represses basal and GnRH-induced transcription of [italic]Fshb[/italic] using a luciferase reporter assay in LbT2 cells. The FOXO1 DNA binding domain (DBD) is necessary for suppression of [italic]Fshb[/italic] transcription, as DBD deletion or mutation relieves transcription suppression. We have mapped FOXO1 suppression to the proximal region of the [italic]Fshb[/italic] promoter. Currently, we are investigating which elements within the proximal promoter and transcription factors mediate FOXO1 suppressive effects. In summary, our data indicate that FOXO1 activity is important in the regulation of [italic]Fshb[/italic] synthesis, and thereby FOXO1 may function as a sensor to couple metabolic signals to gonadotropin synthesis.[br][br]Sources of Research Support: NIH Grants T32 HD720329 to DS; R01 HD067448, K01 DK080467, P30 DK063491 (P [amp] F Award), and U54 HD012303 awarded to VGT.[br][br]Nothing to Disclose: DS, DJA, VGT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 420 73 761 SAT-683 PO19-02 Saturday 700 2012


701 ENDO12L_SAT-684 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) A Potentially Critical Role for the CHD7 Gene in the Ontogeny of GnRH Neurons as Evidenced by Its Role in the Human Disease Model of Isolated GnRH Deficiency (IGD) Nada Al Tassan, Eleni Asimacopoulos, Cassandra Buck, Jennifer Kaina, Lacey Plummer, Ravikumar Balasubramanian, Jin Ho Choi, Richard Quinton, Konstantina Stankovic, Dorota Monies, Fowzan Alkuraya, Brian Meyer, William F Crowley Massachusetts General Hospital, Boston, MA; Massachusetts Eye and Ear Infirmary, Boston, MA; King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia; Royal Victoria Infirmary, Newcastle-upon-Tyne, UK Background: CHARGE is a congenital disorder in which mutations across the 37 exons of CHD7 cause severe developmental defects in several end-organs and the GnRH neurons. Recently, several patients were discovered with IGD who also harbor mutations in CHD7. Also, we reported that oligogenicity occurs in [sim]30% of patients with IGD accounting for much of the varied expressivity and incomplete penetrance observed in this condition.[br]Objectives: 1) To define the incidence, phenotypes (where available), [amp] genotypes of CHD7 mutations in 588 IGD patients; 2) to compare these IGD-associated CHD7 mutations with those in CHARGE patients.[br]Results:[br]Incidence: Of unselected IGD patients, 6% harbor rare CHD7 sequence variants not found in the 1000 Genome Project and scored as pathogenic in 1/4 genetic prediction programs.[br]Phenotypes: Only half of the IGD patients with CHD7 mutations exhibited defects in olfaction. Isolated CHARGE features occur sporadically in these IGD/CHD7+ cases, including hearing defects [5], developmental delay [3], short stature [2], and 1 each with choanal atresia, CV abnormalities, facial asymmetry, and cleft lip/palate. No coloboma was seen.[br]Genotypes: Similar to CHARGE, IGD-associated mutations in CHD7 were equally dispersed across its 37 exons without a mutational hot spot. However, in contrast to CHARGE, most IGD-associated CHD7 mutations (36/37) were milder missense mutations. Second mutations were documented in 13 other genes known to cause IGD, i.e. an incidence of 11/37 [apos]second hits[apos] ([sim]30%): the FGF signaling pathway (5/11), GNRHR (3/11) and KAL1 (2/11) were commonest.[br]Conclusions: 1) The genetic spectrum of CHD7 mutations in IGD is similar in distribution but milder in nature than those causing CHARGE; 2) The range of phenotypic variability associated with these milder mutations in IGD is narrower and less severe than in CHARGE; 3) Oligogenicity is a prominent feature of the genetic architecture of IGD associated CHD7 mutations; 4) The occurrence of KS and nIHH suggests that CHD7 functions in both the neurodevelopmental and neuroendocrine pathways of GnRH neurons; 5) The selective sensitivity of the GnRH neurons to milder CHD7 mutations suggests an especially critical role for CHD7 in their ontogeny.[br][br]Nothing to Disclose: NAT, EA, CB, JK, LP, RB, JHC, RQ, KS, DM, FA, BM, WFC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1999 73 762 SAT-684 PO19-02 Saturday 701 2012


702 ENDO12L_SAT-685 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) A Novel [italic]FGFR1[/italic] Gene Mutation Causing Kallmann Syndrome Dina Belachew, Mark A Sperling Children[apos]s Hospital of Pittsburgh of UPMC, Pittsburgh, PA [bold]Background:[/bold][br]Kallmann syndrome (KS) is characterized by a combination of hypogonadotrophic hypogonadism and one or more nongonadal congenital abnormality, including anosmia, color blindness, midline facial abnormalities, urogenital tract abnormalities, synkinesis and neurosensory hearing loss. Failure of proper migration of olfactory nerves leads to agenesis of the olfactory bulbs and lack of penetration of GnRH neurons into the hypothalamus. The two major forms of KS are X-linked recessive KAL1, and autosomal dominant KAL2, due to mutations of [italic]FGFR1[/italic] on chromosome 8. Many mutations in [italic]KAL1[/italic] have been reported, but there are fewer reported [italic]FGFR1[/italic] mutations with variety of phenotypes.[br][bold]Clinical Case:[/bold][br]A 13-year-10-month old male presented with anosmia and pubertal delay. His past medical history included bilateral cryptorchidism and bilateral cleft lip and palate status, each surgically corrected. Clinical examination showed Tanner III pubic hair but Tanner I testicular and phallic development. Family history was negative for anosmia or pubertal delay in other known family members. Testosterone, FSH and LH levels were prepubertal and very low.[br]Genotyping for KS in the proband revealed a previously unreported heterozygous deletion of c.286_288delTCC in the FGFR1 gene. This in-frame deletion, not previously reported as a mutation or polymorphism, results in the removal of a highly conserved serine residue at codon 96.[br][bold]Conclusions:[/bold][br]To date, no in-frame deletions have been reported as disease-causing mutations in FGFR1 gene. Our patient[apos]s presentation is clinically consistent with Kallmann syndrome therefore leading us to consider that this is a novel mutation. To define this mutation, the family is undergoing further genetic tests, with planned functional in vitro studies to follow.[br][br]Nothing to Disclose: DB, MAS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2078 73 763 SAT-685 PO19-02 Saturday 702 2012


703 ENDO12L_SAT-686 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Identification of a New Gene Involved in Idiopathic Hypogonadotropic Hypogonadism and Kallmann Syndrome by Whole Exome Sequencing Hyung-Goo Kim, Mingyan Fang, Bin Wu, Lawrence C Layman Georgia Health Sciences University, Augusta, GA; Beijing Genomics Institute, Shenzhen, China Whole exome sequencing (WES) has been proven to be a productive strategy to identify new genes involved in human developmental disorders. However, in many diseases this methodology has only limited application since pooling of data is not possible due to genetic heterogeneity. Kallmann syndrome (KS) is characterized by gonadotropin-releasing hormone (GnRH) deficiency with hypogonadotropic hypogonadism and delayed puberty and impaired sense of smell. At least 17 genes have been found to cause IHH/KS, but these only account for about one-third of all cases and digenic causes have only been identified in a minority of families. Despite the significant advances in human molecular genetics of the past two decades, the genetic etiology for most IHH and KS patients remains unknown. To evaluate the efficiency of this WES strategy with a Mendelian disorder of unknown cause, we sought to identify a gene for IHH/KS in a family with multiple affected members in which no mutation has been identified. In this autosomal dominant family, DNA from two patients with normosmic IHH and two with KS were subjected to WES. Exon capture and sequencing of all coding regions of the human genome to a mean coverage of 30X was performed using 90 pair-ends sequencing with the Agilent SureSelect Human All Exon Kit 38M. Emphasis was placed on non-synonymous variants, splice acceptor and donor site variants, and short insertion or deletions of coding region as likely pathogenic mutations. Filtering against dbSNP129, the 1000 Genomes Project, Hapmap8, and Yanhuang databases to exclude common benign sequence variants reduced the pool of candidate genes to 287 under our dominant model in this family, identifying multiple unknown missense and nonsense variants in 39 genes as well as unknown indels in 248 genes. We hypothesized that all affected members will share the same genetic variant of a novel causative gene which will not be shared with unaffected members. Sanger sequencing is currently underway to confirm these sequence changes, and is likely to result in the identification of a causative gene underlying monogenic IHH/KS in this family.[br][br]Sources of Research Support: LCL funded by HD33004 by NICHD.[br][br]Nothing to Disclose: H-GK, MF, BW, LCL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2331 73 764 SAT-686 PO19-02 Saturday 703 2012


704 ENDO12L_SAT-687 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Mutation Analysis of EDG2 (LPAR1) and GPR26 in the Patients with Precocious Puberty Hiromi Nishimoto, Hyung-Goo Kim, Ana C Latronico, Lawrence C Layman Georgia Health Sciences University, Augusta, GA; University of S[atilde]o Paulo, S[atilde]o Paulo, Brazil Timing of pubertal maturation should be commensurate with intellectual development in order for individuals to enter into appropriate social relationships with their peers. In addition, significant pathology may be present in children who initiated sexual development too early. Therefore, the identification of genetic causes for precocious puberty is important. In the present study, we sought to identify disease genes involved in precocious puberty using reported balanced translocations from the individuals affected with precocious puberty. De novo chromosome translocations have been most widely used for the mapping and cloning of disease genes, in which translocation breakpoints provided important information about the gene location. We considered two genes as likely candidate genes for precocious puberty based on their disruption or dysregulation at breakpoints of balanced translocations. The EDG2 (LPAR1) gene localized to 9q31.3 has been reported near the breakpoint in two independent balanced translocations. In a female with macromastia and a 46, XX, t(1;9)(q41;q31.3), EDG2 is located 15 Kb proximal from the 9q31.3 breakpoint. Breast overdevelopment manifested as macromastia could be considered as an exaggerated pubertal phenotype. In another balanced translocation patient with precocious puberty and a t(1;9)(p32.3;q31.3), this gene was disrupted at the 9q31.3 breakpoint. Another candidate GPR26 is contained within a 197 kb BAC (RP11-391M7) spanning the translocation breakpoint at 10q26.13 in a patient with a revised t(6;10)(q21;q26.13), inv(6)(p24.2;q21), who has a precocious puberty as a partial phenotype. Therefore, the coding exons and their immediately flanking intronic regions were subjected to PCR-based DNA sequencing in a cohort of 71 precocious puberty patients. However, no pathogenic mutations were identified in either of these two genes. Our findings indicate that these two genes are unlikely to make a substantial contribution to the pathogenesis of precocious puberty, at least in our subset of patients screened. Thus further investigation of genes disrupted at other breakpoint regions of these three balanced translocation patients is warranted to identify a new causative gene for precocious puberty.[br][br]Sources of Research Support: LCL funded by HD33004 by NICHD.[br][br]Nothing to Disclose: HN, H-GK, ACL, LCL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2262 73 765 SAT-687 PO19-02 Saturday 704 2012


705 ENDO12L_SAT-688 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Cetrorelix, a Gonadotropin-Releasing Hormone, Reduces Disease Activity and Tumor Necrosis Factor-Alpha in Rheumatoid Arthritis: A Proof-of-Concept, Double-Blind, Parallel Group Randomized Controlled Trial (AGRA Trial) Anita Kass, Hans Christian Gulseth, Ivana Hollan, Peter Torjesen, Oystein Forre Betanien Hospital, Skien, Norway; Lillehammer Hospital, Lillehammer, Norway; University of Oslo, Oslo, Norway; Oslo University Hospital, Oslo, Norway [bold]Background[/bold][bold]: [/bold]Effects of gonadotropin-releasing hormone (GnRH) in immune system regulation have been described previously, but never been investigated in humans (1). We aimed to examine whether cetrorelix, a GnRH antagonist, which rapidly decreases luteinising hormone (LH) and follicle-stimulating hormone (FSH), can give a beneficial clinical response in rheumatoid arthritis (RA) patients.[br][bold]Methods[/bold][bold]:[/bold] 99 patients with active RA on stable disease modifying drugs, attending a Norwegian rheumatology outpatients department were randomized 1:1 to receive daily s.c. injections of cetrorelix, (5mg days 1-3, 3mg days 4-5 to achieve a rapid substantial reduction in LH [amp] FSH) or placebo (PBO). Block randomization was carried out by a central interactive computerised system, stratified for sex. The primary endpoint was the change in disease activity score (DAS28) score by day 5 (when LH [amp] FSH levels are lowest) using ANCOVA. Secondary endpoints included the change in TNF-[alpha], ACR (American College of Rheumatology) responses, and proportion of patients in remission. Analysis was by intention to treat. Patients were followed up on day 10 and 15. This study is registered with ClinicalTrials.gov, number NCT00667758.[br][bold]Findings[/bold][bold]: [/bold]Although not reaching statistical significance, the change in DAS28 by day 5 was greater in the cetrorelix group vs. PBO (-0.82 vs. -0.57 [-0.25, 95%CI:-0.57, 0.04] p=0.091). In the cetrorelix group vs. PBO, LH [amp] FSH were reduced (p[lt]0.0001), and TNF-[alpha] was reduced (-25% vs. 13% [diff -37%, 95%CI: -64%, -10%] p=0.008). The proportion of ACR20 responders by day 5 was higher in the cetrorelix group compared to PBO (40% vs. 18%, p=0.015). A higher proportion of patients achieved DAS28 remission (13% vs. 0% p=0.009) and swollen joint count=0 (21% vs. 6% p=0.028) in the cetrorelix group. Occurrence of AEs between groups was similar.[br][bold]Interpretation[/bold][bold]:[/bold] This novel protocol was designed as an exploratory proof-of-concept investigation of the first signals of safety and efficacy of cetrorelix in RA. The study did show that antagonising GnRH improves signs and symptoms of RA and lowers TNF-[alpha] safely in the short term. This study also suggests important interactions between GnRH and the immune system. Thus these outcomes can inform the design of a definitive larger study of longer duration on the efficacy and safety of GnRH antagonists in RA.[br][br](1) Chen A et al. The neuropeptides GnRH-II and GnRH-I are produced by human T cells and trigger laminin receptor gene expression, adhesion, chemotaxis and homing to specific organs. Nat Med 2002; 8: 1421.[br][br]Sources of Research Support: Norwegian Health and Rehabilitation Organisation.[br][br]Nothing to Disclose: AK, HCG, IH, PT, OF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2278 73 766 SAT-688 PO19-02 Saturday 705 2012


706 ENDO12L_SAT-689 POSTER SESSION: Regulation of GnRH, GnRH Signalling [amp] Gonadotropins (1:30 PM-3:30 PM) Effects of Growth Hormone Administration on LH Secretion and Serum Concentrations of Gonadal Steroids in Healthy Older Men and Women Shannon D Sullivan, Ranganath Muniyappa, Johannes D Veldhuis, S Mitchell Harman, Marc R Blackman Georgetown University and Washington Hospital Center, Washington, DC; National Institutes of Health, Bethesda, MD; Mayo Clinical Research Center, Rochester, NY; National Institute on Aging, National Institutes of Health, Baltimore, MD; Kronos Longevity Research Institute, Phoenix, AZ; Phoenix VA Health Care System, Phoenix, AZ; Johns Hopkins University School of Medicine, Baltimore, MD; Veterans Affairs Medical Center, Washington, DC [bold]Background: [/bold]Interactions between the somatotropic and hypothalamic-pituitary-gonadal (HPG) axes are well known. Pituitary gonadotrophs express IGF-I receptors; however, the effects of GH and IGF-I on LH secretion remain uncertain. In young individuals, GH contributes to regulation of both puberty and fertility via HPG axis stimulation. Aging is associated with decreased production of GH and sex steroids (SS), and increased sex hormone binding globulin (SHBG). In this study, we examined the effects of 6 months of GH administration on LH secretory dynamics and SS in healthy older men and women.[br][bold]Objective[/bold]: To determine effects of GH supplementation on LH secretion, SS, and SHBG in healthy older men and women.[br][bold]Methods: [/bold]AM concentrations of LH, IGF-I, total (TT) and free testosterone (fT), estradiol (E2), and SHBG, and nocturnal LH secretory dynamics (2000h-0800h, q20 min sampling) were measured before and after 26 weeks of administration of GH (20 ug/kg sc 3x/week, n=11M, 12W) or placebo (n=12M, 12W) in healthy, older (65-88 yr) individuals with low-normal to mildly decreased IGF-I levels. SHBG and LH were measured by immunoradiometric assay; TT, E2, and IGF-I by RIA; and fT by equilibrium dialysis. LH secretory parameters (frequency, burst mass, pulsatile production rate, and integrated LH secretion) were analyzed by multiparameter deconvolution and orderliness of secretion was assessed by use of the approximate entropy (ApEn) algorithm.[br][bold]Results[/bold]: At baseline, indices of LH secretion (frequency, mass per burst, pulsatile production rate) were inversely (P [lt]0.05) related to IGF-I, but not to mean nocturnal serum GH concentrations. As expected, GH administration significantly increased serum IGF-I. In contrast, GH administration exerted no significant effects on LH secretory dynamics, including ApEn, or concentrations of SS (TT, fT, or E2) or SHBG in older women or men.[br][bold]Conclusions[/bold]: Thus, GH appears to exert little or no significant effect on SS, SHBG, or dynamics of LH secretion in healthy older individuals despite normalization of low-normal baseline IGF-1 levels, in contrast to its stimulatory effect on the HPG axis in younger persons. These data suggest a dissociation in the relationship between somatotropic and HPG axis function with aging that may contribute to the initiation and/or progression of menopause and andropause.[br][br]Nothing to Disclose: SDS, RM, JDV, SMH, MRB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 202 73 767 SAT-689 PO19-02 Saturday 706 2012


707 ENDO12L_SAT-690 POSTER SESSION: GH Deficiency: Diagnosis, Prognosis [amp] Therapy (1:30 PM-3:30 PM) Growth Hormone Deficiency in Veterans with Traumatic Brain Injury: An Atlanta VAMC Pilot Study Adriana G Ioachimescu, Benjamin M Hampstead, Elisabeth A Burgess, Anna B Moore, Lawrence S Phillips Emory School of Medicine, Atlanta, GA; Emory School of Medicine, Atlanta, GA Traumatic brain injury (TBI) has been recognized as a cause of growth hormone (GH) deficiency in civilians, but there have been no studies in veterans returned from combat - where TBI results largely from exposure to shock waves from explosions.[br]We studied 20 men with mild TBI incurred during combat. Endocrine evaluation included glucagon GH stimulation (deficiency if peak [lt] 3 ng/mL), IGF-1, free T4, TSH and cortisol measurement. Subjects with a.m. serum cortisol [lt] 10 [micro]g/dL underwent ACTH stimulation tests. Neuropsychological evaluation included assessment of cognitive and emotional status as well as quality of life. Due to the pilot nature of the study, we used Cohen[apos]s d test to examine differences between the GH-sufficient and deficient groups; medium (d[gt]0.5) and large (d[ge]0.8) effect sizes were considered.[br]The mean age was 33.7[plusmn]7.8 yr. The response to glucagon was subnormal in 5 subjects (25%), and IGF-1 was low in one of these subjects. All subjects had normal thyroid and normal cortisol status. Sixteen subjects provided sufficient effort to deem the neuropsychological results valid (12 GH-sufficient, 4 GH-deficient). The 2 groups performed comparably on measures of simple and complex attention, learning, and memory. However, the GH-deficient group demonstrated more intrusive errors during memory testing (d=1.48), and performed relatively worse on measures of inhibitory control (d=0.92 [ndash] 1.2). Depression severity was worse in the GH-deficient group (d=0.80), but no differences were suggested in the levels of fatigue or post-traumatic stress disorder. Finally, GH-deficient patients reported a lower quality of life relative to the GH-sufficient group based on AGHDA questionnaire scores (d=0.64). In summary, our pilot study indicates a prevalence of 25% of GH deficiency in veterans with mild TBI from exposure during combat. GH-deficient subjects exhibited more severe depression and lower quality of life. The neuropsychological findings raise the possibility that GH deficiency is also associated with self-monitoring and inhibitory control deficits. Further studies are needed to confirm these findings, to elucidate the nature of the executive dysfunction and emotional perturbations, and to determine whether these problems and rehabilitation can be improved with GH replacement.[br][br]Sources of Research Support: Novo Nordisk (Investigator Initiated Study).[br][br]Nothing to Disclose: AGI, BMH, EAB, ABM, LSP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 538 74 768 SAT-690 PO21-01 Saturday 707 2012


708 ENDO12L_SAT-691 POSTER SESSION: GH Deficiency: Diagnosis, Prognosis [amp] Therapy (1:30 PM-3:30 PM) Anterior Pituitary Dysfunction in Veterans with Blast Exposure [mdash] A Pilot Study Amy D Anderson, Jeffrey Taylor, Shane Mcnamee, Robert A Adler University of Virginia Health System, Charlottesville, VA; Virginia Commonwealth University Health System, Richmond, VA; McGuire Veterans Affairs Medical Center, Richmond, VA; McGuire Veterans Affairs Medical Center, Richmond, VA Traumatic brain injury (TBI) has been reported to cause pituitary dysfunction in up to 30% of patients. In recent wars, in addition to penetrating wounds and other typical head injuries, blast exposure has become a common cause of TBI. This pilot study was done to determine if blast exposure was associated with disorders of pituitary function. A convenience sample of 26 veterans with self-reported blast injury was studied for evidence of anterior pituitary dysfunction. None were taking medications known to affect pituitary function, including opiate analgesics. All denied using anabolic agents. The subjects underwent a short ACTH stimulation test and 4 hour growth hormone response to glucagon test on 2 separate days. Fasting basal hormone levels were also obtained. Overall severity of the injury was assessed by a blinded physiatrist. All subjects were male, ages 22 to 44, median 32. All were ambulatory. Two had total testosterone levels [lt] 200 ng/dL, 4 between 200 and 300, and 7 between 300 and 400, all below expected levels for age (1). One man had elevated serum testosterone and sex hormone binding globulin. LH and FSH were not elevated, and prolactin was normal except for a mild elevation in one subject. In all subjects the cortisol response to 250 [mu]g of intramuscular ACTH was normal as was the free thyroxine. IGF-I levels were normal in all but one subject, but the growth hormone response to intramuscular glucagon was [lt] 5 [mu]g/L in 15 ([lt] 3 [mu]g/L in 9). The one subject with a low IGF-I also had a peak growth hormone level of 0.2 [mu]g/L. Of those men with blast sustained TBI, all but 2 were classified as [ldquo]mild[rdquo] TBI. In addition, the great majority of the subjects also carried the diagnosis of post-traumatic stress disorder (PTSD). From this pilot study, we conclude that low serum testosterone levels without gonadotropin elevation suggesting secondary hypogonadism are common in men with blast exposure, but the contributions of the injury and PTSD will require further study. The deficient growth hormone response to glucagon despite normal IGF-I also requires further study and testing with other secretagogues.[br][br]Bhasin S, et al, J Clin Endocrinol Metabl 2011; 96:2430.[br][br]Sources of Research Support: Genentech.[br][br]Nothing to Disclose: ADA, JT, SM, RAA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 233 74 769 SAT-691 PO21-01 Saturday 708 2012


709 ENDO12L_SAT-692 POSTER SESSION: GH Deficiency: Diagnosis, Prognosis [amp] Therapy (1:30 PM-3:30 PM) Reevaluation of Stimulated Growth Hormone (GH) Release in Adult Thalassemic Patients Previously Classified as Having Normal GH Secretion, Partial GH Deficiency (GHD) or Severe GHD Agnese Cattaneo, Leila Danesi, Massimo Scacchi, Raffaella Radin, Giovanna Sciortino, Emanuela D[apos]Angelo, Nadia Mirra, Laura Zanaboni, Elena Cassinerio, Maria Domenica Cappellini, Francesco Cavagnini, Luca Persani University of Milan, Milan, Italy; Ospedale San Luca IRCCS, Istituto Auxologico Italiano, Milan, Italy; Ospedale San Luca IRCCS, Istituto Auxologico Italiano, Milan, Italy; Fondazione Policlinico Mangiagalli Regina Elena, Milan, Italy; University of Milan, Milan, Italy; Istituto Auxologico Italiano, Milan, Italy In previous studies we demonstrated, by the single test GHRH + arginine, severe and partial GH deficiency (GHD) in 22-25 and 17-19%, respectively, of adult thalassemic patients (1,2). However, the need to perform two provocative tests for the correct diagnosis of GHD in thalassemic adults has been very recently suggested (3,4). In non thalassemic adults undergone pituitary surgery, a progression to severe GHD (sGHD) has been reported in up to 40% of patients with partial GHD (pGHD) (5). Theoretically, this progression might be favoured in thalassemia by the increasing iron overload. Based on these considerations, we elected to reassess the GH secretory reserve in thalassemic adults at two years since the original testing. [bold]Patients and methods[/bold]. The study group (20 men and 17 women, aged 19-48 years) included 17 thalassemic adults previously displaying normal GH responses to GHRH + arginine, 10 patients previously diagnosed with pGHD and 10 previously diagnosed as having sGHD. Two years after the first assessment, the GH response to GHRH (1 mcg/kg b.w.) + arginine (0.5 g/kg b.w.) was reevaluated in each patient. [bold]Results[/bold]. Upon retesting 17.6% of the patients originally classified as GH sufficient were diagnosed with pGHD and another 17.6% with sGHD. Partial GHD progressed to sGHD in 40% of the cases and improved to normal GH secretion in another 40%. Finally, sGHD was confirmed only in 20% of the patients with this original diagnosis: surprisingly, stimulated GH release was improved to pGHD in 50% of them and even normalized in another 30%. [bold]Conclusion[/bold]. When using the single GHRH + arginine test, the progression from pGHD to sGHD was observed in a proportion of thalassemic adults similar to that reported in non thalassemic hypopituitary patients. However, the high percentage of subjects in whom sGHD was not confirmed upon retesting indicates that a single provocative test may not be sufficient to establish a correct diagnosis of GHD in these peculiar patients.[br][br](1)Scacchi et al., Clin Endocrinol 2007; 67:790. (2)Scacchi et al., Clin Endocrinol 2008; 69:202. (3)Pincelli et al., Pediatr Endocrinol Rev 2011; 8(Suppl 2):284. (4)De Sanctis et al., Pediatr Endocrinol Rev 2011; 8(Suppl 2):290. (5)Colao et al., J Clin Endocrinol Metab 2006; 91:2191.[br][br]Nothing to Disclose: AC, LD, MS, RR, GS, ED, NM, LZ, EC, MDC, FC, LP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 242 74 770 SAT-692 PO21-01 Saturday 709 2012


710 ENDO12L_SAT-693 POSTER SESSION: GH Deficiency: Diagnosis, Prognosis [amp] Therapy (1:30 PM-3:30 PM) Continuous Positive Airway Pressure (CPAP) Improves Hyposomatotropism in Men with Obstructive Sleep Apnea (OSA): A Randomized Sham-Controlled 12 Week Study Camilla M Hoyos, Daniel M Keenan, Johannes D Veldhuis, Peter Y Liu Woolcock Institute of Medical Reserach, Sydney, Australia; University of Sydney, Sydney, Australia; University of Virginia, Charlottesville, VA; Mayo Clinic College of Medicine, Rochester, MN; Royal Prince Alfred Hospital, Sydney, Australia Background: OSA is a common condition that is characterised by intermittent hypoxia, which disrupts sleep and is associated with reduced growth hormone (GH) secretion. Hyposomatotropism contributes to metabolic ill-health in men with OSA, but the only randomised sham-controlled trial reported no change morning IGF-1 after 4 weeks of CPAP.[br]Methods: 65 CPAP na[iuml]ve men with moderate to severe OSA (age=49[plusmn]12y, apnea hypopnea index (AHI)=40[plusmn]18 events/h, BMI=31[plusmn]5 kg/m2, IGF-1 =20.4[plusmn]6.3 nmol/L) were randomised in a 12-week double blind sham-controlled parallel group study, to receive either active (n=34) or sham (n=31) CPAP. OSA was measured by polysomnography at week 0 and 12 and fasting morning glucose, insulin, C-peptide and IGF-1 blood levels at 0, 6 and 12 weeks in all men. GH was also measured in a subgroup of 18 men (active n=11, sham n=7) every 10-minutes from 10PM until 6AM (49 samples) at week 12 while sleep architecture (960 epochs) was simultaneously assessed. GH secretion was determined by mathematical deconvolution. Data are mean [plusmn]SD.[br]Results: AHI, BMI and IGF-1 (all p[gt]0.16) were not different at baseline between groups. As expected 12 weeks of active, compared to sham, CPAP reversed OSA by 33 events/hr (p[lt]0.0001) and increased slow wave sleep by 7% (p=0.05). Mixed model analyses showed that active, compared with sham, CPAP increased IGF-1 by 2.94 nmol/L (p=0.006) at 12 weeks, but not at 6 weeks (0.83 nmol/L, p=0.4). Changes in fasting glucose, insulin, C-peptide/glucose and HOMA insulin sensitivity were not different between groups at 6 or 12 weeks (all p[gt]0.15). In the subgroup of 18 men that underwent overnight blood sampling at 12 weeks, total (35.0[plusmn]28.0 versus 8.5[plusmn]7.3 ng/ml/8h, p=0.001) and pulsatile (32.3[plusmn]27.7 vs 6.8[plusmn]6.8 ng/ml/8h, p=0.002) GH secretion, mean GH concentration (1.2[plusmn]1.0 vs 0.3[plusmn]0.3 ng/ml, p=0.002) and mass of GH secreted per pulse (8.7[plusmn]9.0 vs 2.2[plusmn]2.3 ng/ml, p=0.01) were all significantly higher in active compared with sham CPAP users, respectively. The pulse frequency (p=0.11), interpulse regularity (p=0.6) and GH regularity (p=0.78) were not different between groups.[br]Conclusions: Twelve weeks of CPAP increases total and pulsatile GH secretion, secretory burst mass and pulse frequency. Six weeks of CPAP may be insufficient, whereas 3 months is sufficient, to increase IGF-1 but not alter insulin action. Persistence with CPAP is required to improve abnormalities in GH secretion and consequent metabolic effects associated with OSA.[br][br]Sources of Research Support: The National Health and Medical Research Council of Australia (NHMRC) through a project grant (512498), a Centre for Clinical Research Excellence in Interdisciplinary Sleep Health (571421) and fellowships to CH and PYL (512057 and 1025248, respectively). Sham machines were provided by Phillips Respironics.[br][br]Nothing to Disclose: CMH, DMK, JDV, PYL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1283 74 771 SAT-693 PO21-01 Saturday 710 2012


711 ENDO12L_SAT-694 POSTER SESSION: GH Deficiency: Diagnosis, Prognosis [amp] Therapy (1:30 PM-3:30 PM) Predictors of Change in Bone Mineral Density (BMD) in Response to Growth Hormone (GH) Replacement for 4 Years in GH-Deficient (GHD) Adults [mdash] A KIMS (Pfizer International Metabolic Database) Analysis Nicholas A Tritos, Amir H Hamrahian, Donna King, Susan L Greenspan, David M Cook, Peter J Jonsson, Maria Koltowska-Haggstrom, Beverly MK Biller Massachusetts General Hospital and Harvard Medical School, Boston, MA; Cleveland Clinic, Cleveland, OH; Pfizer, Inc, New York, NY; University of Pittsburgh, Pittsburgh, PA; Oregon Health and Science University, Portland, OR; Pfizer Endocrine Care, Sollentuna, Sweden GH replacement may increase BMD in GHD adults. To investigate factors predictive of BMD response, the KIMS database was searched using these criteria: stringently defined GHD of adult onset, true GH na[iuml]ve status at study entry, and BMD measured in the posterior-anterior lumbar spine (LS) and/or femoral neck (FN) on the same densitometer for each subject, both at study entry and after 4 years of GH replacement.[br]The search identified 258 subjects, including 130 women and 128 men. Study population characteristics were [median (10[sup]th[/sup] percentile, 90[sup]th[/sup] percentile)]: age at study entry: 48.9 years (33.8, 66.5); number of additional pituitary hormone deficits (% subjects): isolated GHD (7.8%); 1 (17.0%); 2 (13.2%); 3 (46.1%); 4 (15.9%); gonadotropin deficiency and corresponding sex steroid replacement status (% subjects): deficient but unreplaced (17.4%); deficient and replaced (64.8%); endogenously sufficient (17.8%); serum insulin-like growth factor-I standard deviation scores (at baseline): -1.6 (-4.1, -0.1); GH dose at 4 years: 0.4 mg/day (0.2, 0.8); baseline LS Z score: -0.5 (-2.0, 1.5); baseline FN Z score: -0.3 (-1.5, 1.1).[br]After 4 years of GH replacement, there was a 6.7% increase in LS BMD in men (P=0.0001) and a 3.3% increase in LS BMD in women (P=0.0003) over baseline [3.1% increase (P=0.0117) in women [lt] 50 yr and 3.5% increase (P=0.0074) in women [gt] 50 yr]. On univariate analysis, the increase in LS BMD was significantly greater in men than women (P=0.0004). There was no change in FN BMD in men or women over baseline [with positive % FN BMD trends in women [lt] 50 yr; the difference in % FN BMD change between women [lt] 50 yr and [gt] 50 yr being significant (P=0.0283)].[br]On GH replacement, there was a 3.0% increase in LS BMD in sex steroid deficient/unreplaced (P=0.004), a 5.9% increase in LS BMD in deficient/replaced (P=0.0001) and a 2.0% increase in LS BMD in sufficient (P=0.0445) subjects. On multivariate analysis, sex steroid replacement (r=0.29, P=0.0051) and older age at pituitary disease onset (r=0.24, P=0.0145) were independent predictors of percent increase in LS BMD.[br]In conclusion, GH replacement for 4 years led to a significant increase in LS BMD in adult GHD. Sex steroid replacement and older age at pituitary disease onset were independently associated with a greater increase in LS BMD after 4 years of GH replacement. Sex steroid replacement should be considered in appropriate hypopituitary patients and may improve skeletal benefits of GH replacement.[br][br]Disclosures: NAT: Consultant, Pfizer, Inc. AHH: Consultant, Pfizer, Inc.; Speaker, Pfizer, Inc.; Principal Investigator, Eli Lilly & Company, Novo Nordisk. DK: Employee, Pfizer, Inc. DMC: Speaker, Pfizer, Inc., Novo Nordisk. PJJ: Employee, Pfizer, Inc. MK-H: Employee, Pfizer, Inc. BMKB: Consultant, Pfizer, Inc., Novo Nordisk; Principal Investigator, Pfizer, Inc., Novo Nordisk, Serono. Nothing to Disclose: SLG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 776 74 772 SAT-694 PO21-01 Saturday 711 2012


712 ENDO12L_SAT-695 POSTER SESSION: GH Deficiency: Diagnosis, Prognosis [amp] Therapy (1:30 PM-3:30 PM) Arrest of Atherosclerosis Progression after Interruption of GH Replacement in Adults with Congenital Isolated Growth Hormone Deficiency Vanessa P Araujo, Manuel H Aguiar-Oliveira, Joselina ML Oliveira, Carla RP Oliveira, Isabella MPA Britto, Roberto Ximenes, Jose AS Barreto-Filho, Rafael A Meneguz-Moreno, Rossana MC Pereira, Eugenia HO Valenca, Luiz A Oliveira-Neto, Taisa AR Vicente, Amanda Blackford, Roberto Salvatori Federal University of Sergipe, Aracaju, Brazil; Johns Hopkins University, Baltimore, MD; Johns Hopkins University, Baltimore, MD [bold]Background: [/bold]GH replacement therapy (GHRT) in adult-onset GH deficiency (AOGHD) reduces cholesterol level and carotid intima media thickness (IMT) and increases myocardial mass, with improvement of both systolic and diastolic function. These observations have reinforced the use of GHRT in AOGHD. Conversely, we have previously reported that in 20 adults with lifetime congenital, severe, and previously untreated isolated GH deficiency (IGHD) due to a homozygous mutation in GHRH receptor gene, a 6-month treatment with bi-monthly depot GH (despite positive effect on cholesterol and body composition) increased carotid IMT and caused the development of atherosclerotic plaques. In addition, GHRT increased left ventricular mass index (LVMI), posterior wall and septal thickness, and ejection fraction (1). Such effects persisted 12 months after treatment discontinuation (12 months wash out, 12mo).[br][bold]Methods:[/bold] We have now studied the cardiovascular status of 19 of these 20 subjects (one had died for pneumonia) 60 months after completion of therapy (60 months washout 60mo).[br][bold]Results:[/bold] Serum cholesterol did not change during extended wash out, while IMT reduced significantly from 12mo to 60mo, returning to baseline (pre-GH therapy) value. The number of individuals with plaques was similar at 12mo and 60mo, remaining significantly higher than pre-therapy. LVMI, relative posterior wall thickness, and septum thickness did not change between 12mo and 60mo, but absolute posterior wall increased from 12mo to 60mo. Systolic function, evaluated by ejection fraction and shortening fraction, was reduced at 60mo in comparison to 12mo, returning to baseline levels. The E/A wave ratio (expression of diastolic function) decreased at 60mo compared to both 12mo and baseline.[br][bold]Conclusion: [/bold]In adults with lifetime and previously untreated congenital severe IGHD the progression of atherosclerosis caused by 6 months of GHRT stopped after 12 months from discontinuation of therapy. Similarly, systolic function benefits (still present at 12mo) disappeared 4 years later. Conversely, some structural heart changes were permanent.[br][br]1) Oliveira JL et al. J Clin endocrinol Metab 2007 92:4664.[br][br]Sources of Research Support: NIDDK, Genenetech Foundation for Clinical research.[br][br]Disclosures: RS: Advisory Group Member, Novo Nordisk. Nothing to Disclose: VPA, MHA-O, JMLO, CRPO, IMPAB, RX, JASB-F, RAM-M, RMCP, EHOV, LAO-N, TARV, AB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 192 74 773 SAT-695 PO21-01 Saturday 712 2012


713 ENDO12L_SAT-696 POSTER SESSION: GH Deficiency: Diagnosis, Prognosis [amp] Therapy (1:30 PM-3:30 PM) Pregnancies and Live Births in a Large Cohort of Patients on Growth Hormone Replacement Therapy (GHRT): Results from KIMS (Pfizer International Metabolic Database) Greisa Vila, Ann-Charlotte Akerblad, Beverly MK Biller, Maria Koltowska-Haggstrom, Frida Lundgren, Michaela Riedl, Anton Luger Medical University of Vienna, Vienna, Austria; Pfizer Health AB, Sollentuna, Sweden; Massachusetts General Hospital and Harvard Medical School, Boston, MA [underline]Introduction:[/underline] GHRT during conception and pregnancy in women with growth hormone deficiency (GHD) is an off-label treatment. Pregnancy outcomes in these patients have only been evaluated in single center studies with small sample sizes, with conflicting results. To date, there are few data and no guidelines regarding whether to continue or withdraw GHRT during conception and pregnancy. Moreover, there are no data on the effect of paternal GH exposure on the evolution and outcome of a partner[apos]s pregnancy.[br][underline]Methods/design:[/underline] We searched for pregnancy reports in KIMS, a pharmacoepidemiological study of adults with GHD. This database includes information from 4,651 women aged 15 to 50 years (followed for 19,348 patient-years; median 3.10 years) and 7,438 men aged above 16 years (followed for 36,074 patient-years; median 3.89 years).[br][underline]Results:[/underline] A total of 151 pregnancies were reported from 131 patients, of which 116 were in women (age 22 to 41 years) and 15 were in partners of male patients (men[apos]s age 26 to 68 years). Ovulation induction or assisted reproduction treatments were used in 65% of women; (10% of women and one man had discontinued GHRT before conception). GHRT use in pregnant women was as follows: continuation in 24% of the cases, withdrawal in 56% and dose reduction in 2%. Within the cohort that stopped GH during pregnancy, 46% gave birth to healthy babies, 12% reported fetal loss, and outcome is not reported in 42% of the cases. Within the cohort that continued GH during pregnancy, 41% gave birth to healthy babies, 34% reported fetal loss, and outcome is not reported in 24% of the cases. The category [ldquo]fetal loss[rdquo] includes both miscarriages and elective abortions. At the time of abstract submission, birth of 82 healthy children has been confirmed, 20 of them from twin pregnancies and 3 from a triplet pregnancy. Of note, 14 healthy children were born to women who continued GH during conception and pregnancy and 6 with paternal exposure to GH.[br][underline]Conclusions:[/underline] We provide the first demographic data on pregnancy rates in a large cohort of patients receiving GHRT. As might be anticipated given the underlying hypothalamic-pituitary disorders, two-thirds of women were treated to achieve fertility. It appears that in the clinical practice setting, nearly all patients taking GH replacement continue treatment during the time when they seek fertility, and one-fourth continue it during pregnancy. The collection of missing data on pregnancy outcome is ongoing.[br][br]Disclosures: A-CA: Employee, Pfizer, Inc. BMKB: Consultant, Pfizer, Inc., Novo Nordisk; Principal Investigator, Pfizer, Inc., Novo Nordisk, Serono. MK-H: Employee, Pfizer, Inc. FL: Employee, Pfizer, Inc. AL: Consultant, Pfizer, Inc. Nothing to Disclose: GV, MR 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1209 74 774 SAT-696 PO21-01 Saturday 713 2012


714 ENDO12L_SAT-697 POSTER SESSION: GH Deficiency: Diagnosis, Prognosis [amp] Therapy (1:30 PM-3:30 PM) Baseline Clinical Characteristics and Response to 1 Year of GH Replacement in Adults with GH Deficiency (GHD) Due to Childhood-Onset (CO) Craniopharyngioma, CO Idiopathic/Congenital Hypopituitarism, and CO Extrasellar Tumors: Data from KIMS (Pfizer International Metabolic Database) KCJ Yuen, DM Cook, M Koltowska-Haggstrom, JL Fox, PJ Jonsson, ME Geffner, R Abs Oregon Health [amp] Science University, Portland, OR; Pfizer Endocrine Care, Sollentuna, Sweden; Pfizer, Inc, New York, NY; Children[apos]s Hospital Los Angeles, Los Angeles, CA; Antwerp Center for Endocrinology, Antwerp, Belgium [italic]Context:[/italic] Prior studies have shown that CO craniopharyngioma (COCP) patients have poorer clinical outcomes compared to many other causes of hypopituitarism (1). Hypothalamic damage from the tumor itself [amp]/or its treatment (surgery [amp]/or radiotherapy) are likely explanations for the increased mortality. Few studies have examined the characteristics of COCP with GHD [amp] their response to 1-yr of GH replacement. We analyzed data from KIMS, [amp] compared baseline characteristics of adults with GHD due to COCP to those with CO idiopathic/congenital hypopituitarism (COH) [amp] CO extrasellar tumors (COE), [amp] describe their responses to 1-yr of GH replacement.[br][italic]Methods:[/italic] We studied 260 COCP patients (152M, median age 24.4 yr), 498 COH patients (298M, median age 25.8 yr) [amp] 272 COE patients (149M, median age 22.2 yr). All COCP [amp] COE, but not COH, patients underwent surgery [amp]/or radiotherapy.[br][italic]Results:[/italic] At baseline, median duration of GHD differed between the 3 groups (COCP 6.9 vs COH 13.9 vs COE 4.5 yr; P[lt]0.01). Compared to COH [amp] COE patients, COCP patients had higher BMI, waist/hip ratio (WHR), triglycerides (TG), lean body mass (LBM) [amp] fat mass (FM) (all P[lt]0.01), higher prevalence of pituitary deficiencies, visual field defects (all P[lt]0.0001) [amp] diabetes mellitus (P[lt]0.05), [amp] comparable total [amp] LDL cholesterol. Compared to COH patients, COCP patients had lower HDL, higher bone mineral content (BMC) (all P[lt]0.01) [amp] comparable IGF-I SDS. Compared to COE patients, COCP patients had lower IGF-I SDS (P[lt]0.001), [amp] comparable HDL [amp] BMC. After 1-yr of GH replacement, similar changes were evident in BMI, WHR, LBM, FM, fasting glucose, hemoglobin A1c (A1c), total [amp] LDL cholesterol, [amp] BMC in all 3 groups. However, compared with COE patients, GH replacement in COCP patients decreased TG (-0.10 vs 0.10 mmol/L; P[lt]0.05) [amp] LDL (-0.32 vs -0.09 mmol/L; P[lt]0.05) more.[br][italic]Conclusion:[/italic] Compared to COH [amp] COE patients, COCP patients were more obese, had higher prevalence of pituitary deficiencies, visual field defects [amp] dysglycemia, [amp] higher TG that may explain the higher cardiovascular (CV) risk [amp] mortality in these patients. After 1-yr of GH replacement, COCP patients responded with similar changes to COH [amp] COE patients in BMI, WHR, body composition, fasting glucose, A1c, total cholesterol [amp] BMC. Thus, despite having more CV risk factors, the response of COCP patients to the beneficial effects of short-term GH replacement is neither influenced by the underlying baseline characteristics nor by duration of GHD.[br][br]1. Sherlock M, et al. Mortality in patients with pituitary disease. Endocr Rev. 2010; 31:301-342.[br][br]Sources of Research Support: KCJY: Speaker, Pfizer, Inc., Novo Nordisk; Principal Investigator, Pfizer, Inc., Novo Nordisk; Medical Advisory Board, Pfizer, Inc., Novo Nordisk. DMC: Medical Advisory Board, Novo Nordisk; Speaker, Pfizer, Inc., Eli Lilly and Company; Principal Investigator, Pfizer, Inc. MK-H, JLF, PJJ: Employees, Pfizer, Inc. MEG: Medical Advisory Board, Pfizer, Inc.; Consultant: KIGS Strategic Advisory Board; Principal Investigator, Pfizer, Inc. RA: Consultant: KIMS Strategic Advisory Board; Member: KIMS International Board.[br][br]Disclosures: MK-H: Employee, Pfizer, Inc. JLF: Employee, Pfizer, Inc. PJJ: Employee, Pfizer, Inc. Nothing to Disclose: KCJY, DMC, MEG, RA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 658 74 775 SAT-697 PO21-01 Saturday 714 2012


715 ENDO12L_SAT-698 POSTER SESSION: GH Deficiency: Diagnosis, Prognosis [amp] Therapy (1:30 PM-3:30 PM) Quality of Life, Patient Personal Situations, Patient Reported Outcomes and Healthcare Utilization at Baseline and after 1 Year of GH Replacement in Adults with GH Deficiency (GHD) Due to Childhood-Onset (CO) Craniopharyngioma, CO Idiopathic/Congenital Hypopituitarism, and CO Extrasellar Tumors: Data from KIMS (Pfizer International Metabolic Database) KCJ Yuen, DM Cook, M Koltowska-Haggstrom, JL Fox, PJ Jonsson, ME Geffner, R Abs Oregon Health [amp] Science University, Portland, OR; Pfizer Endocrine Care, Sollentuna, Sweden; Pfizer, Inc, New York, NY; Children[apos]s Hospital Los Angeles, Los Angeles, CA; Antwerp Center for Endocrinology, Antwerp, Belgium [italic]Context:[/italic] In CO craniopharyngioma (COCP) patients, hypothalamic damage associated with obesity is often implicated as the cause of poor quality of life (QoL) (1), but few studies have compared the QoL of COCP patients to other causes of CO hypopituitarism. We analyzed data from KIMS, [amp] compared baseline QoL, healthcare utilization, patient personal situations [amp] reported outcomes of adults with GHD due to COCP to those with CO idiopathic/congenital hypopituitarism (COH) [amp] CO extrasellar tumors (COE), [amp] describe responses to 1-yr of GH replacement.[br][italic]Methods:[/italic] We studied 260 COCP patients (152M, median age 24.4 yr), 498 COH patients (298M, median age 25.8 yr) [amp] 272 COE patients (149M, median age 22.2 yr). Healthcare utilization, patient personal situations, reported outcomes, [amp] QoL were assessed by the KIMS Patient Life Situation Forms [amp] QoL-AGHDA questionnaires, respectively. All COCP [amp] COE, but not COH, patients underwent surgery [amp]/or radiotherapy.[br][italic]Results:[/italic] At baseline, there were no differences in marital status, education, employment [amp] number of children among the 3 groups. Most patients did not require assistance with daily living (COCP 74.7%, COH 87.6%, COE 73.3%), whereas in those who did, there were more COCP than COH patients (25.3% vs 12.4%; P[lt]0.001). Compared to COH patients, COCP patients had poorer QoL, higher number of hospital admission days (both P[lt]0.01) [amp] a trend of more doctor visits (P=0.06). Compared to COE patients, COCP had comparable QoL, [amp] number of hospital admission days [amp] doctor visits. After 1-yr of GH replacement, COCP patients reported improvement in QoL but not as much as COE patients (QoL-AGHDA scores -2.0 vs -4.0; P[lt]0.05), while comparable improvements were reported in COH patients. All 3 groups did not report any change in the level [amp] degree of satisfaction of daily activities after 1-yr of GH replacement.[br][italic]Conclusion:[/italic] Our study demonstrates that most patients in the 3 groups lived independently with comparable personal situations, but COCP patients reported poorer QoL [amp] seeked medical attention more frequently than COH, but not COE, patients. In addition, short-term GH replacement improved QoL in all 3 groups but the degree of improvement is less so in COCP compared to COE patients. Thus, poorer QoL in COCP patients may be due to the tumor itself [amp]/or hypothalamic damage from previous surgery [amp]/or radiotherapy, while the lack of subjective satisfaction in daily activities may be due to too short a duration of GH replacement.[br][br]1. Kendall-Taylor P, et al. The clinical, metabolic and endocrine features and the quality of life in adults with childhood-onset craniopharyngioma compared with adult-onset craniopharyngioma. Eur J Endocrinol. 2005; 152:557-567.[br][br]Sources of Research Support: KCJY: Speaker, Pfizer, Inc., Novo Nordisk; Principal Investigator, Pfizer, Inc., Novo Nordisk; Medical Advisory Board, Pfizer, Inc., Novo Nordisk. DMC: Medical Advisory Board, Novo Nordisk; Speaker, Pfizer, Inc., Eli Lilly and Company; Principal Investigator, Pfizer, Inc. MK-H, JLF, PJJ: Employees, Pfizer, Inc. MEG: Medical Advisory Board, Pfizer, Inc.; Consultant: KIGS Strategic Advisory Board; Principal Investigator, Pfizer, Inc. RA: Consultant: KIMS Strategic Advisory Board; Member: KIMS International Board.[br][br]Disclosures: MK-H: Employee, Pfizer, Inc. JLF: Employee, Pfizer, Inc. PJJ: Employee, Pfizer, Inc. Nothing to Disclose: KCJY, DMC, MEG, RA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 659 74 776 SAT-698 PO21-01 Saturday 715 2012


716 ENDO12L_SAT-699 POSTER SESSION: GH Deficiency: Diagnosis, Prognosis [amp] Therapy (1:30 PM-3:30 PM) Mortality Rates in Patients with Childhood and Adult-Onset GH Deficiency Enrolled in the Global Hypopituitary Control and Complications Study (HypoCCS) Heike Jung, Alan G Zimmermann, Daojun Mo, Leslie L Robison, Eva-Marie Erfurth, Steven WJ Lamberts, Shlomo Melmed, Andrea F Attanasio Eli Lilly and Company, Bad Homburg, Germany; St Jude Children[apos]s Research Hospital, Memphis, TN; Lund University, Lund, Sweden; Cedars-Sinai Medical Center, Los Angeles, CA; Erasmus University, Rotterdam, Netherlands; Cascina del Rosone, Agliano Terme, Italy Preliminary data suggested an increased mortality in a French cohort after childhood somatropin treatment compared to the French reference population[sup](1)[/sup]. This cohort included patients with idiopathic-isolated GH-deficiency (GHD), and a similar cohort is also under follow-up for adult GH-replacement in HypoCCS.[br]We therefore assessed all-cause mortality rates (n/1000 person-years (PY), [95% CI], standardized to the age/gender structure of HypoCCS or the reference populations) in 7946 GH-replaced adult HypoCCS patients with available post-baseline information. Overall standardized (general population) mortality rate was 5.8 [4.3, 7.2]/1000 PY in the US HypoCCS cohort compared to 7.6/1000 PY in the US general population[sup](2)[/sup], and 5.2 [4.4, 6.1]/1000 PY in Europe, the rate in comparable populations ranging from 5.1/1000 PY in Italy to 6.4/1000 PY in Belgium[sup]([/sup][sup]3)[/sup].[br]Standardized mortality rates in HypoCCS were not different between childhood onset (CO) and adult onset (AO) GHD-types overall (CO: 3.2 [0.2, 6.2]/1000 PY, N=1528; AO: 5.4 [4.5, 6.3]/1000 PY, N=6418). No difference was seen in rates between patients with CO and AO non-idiopathic GHD (N=6679), the rates being 5.4 [0.2, 10.6]/1000 PY and 5.3 [4.3, 6.2]/1000 PY, respectively. However, in patients with CO and AO idiopathic GHD (N=1267), rates were lower in CO (0/1000 PY) compared to AO (7.0 [3.9, 10.1]/1000 PY). In a subgroup of patients with isolated-idiopathic GHD (N=530) the rates were not different between patients with CO (0/1000 PY) and AO GHD (3.7 [0, 7.8]/1000 PY).[br]Of a total of 174 deaths, 14 occurred in CO patients and only in the non-idiopathic GHD group (acute illness/suicide (7), second/recurrent neoplasm (2), cerebral hemorrhage/carotid artery stenosis (2) or unknown reason (3)).[br]Although the Carel et al study[sup](1)[/sup] raises the important question of increased adult mortality after somatropin treatment in childhood, the present analysis does not confirm an increased mortality in GH-replaced adult patients, either with CO or AO GHD, compared to reference populations. However, there are limitations in our study including the selected population of patients and relatively short follow-up time.[br][br](1) Carel J-C et al., J Clin Endocrinol Metab 2012; Jan 11. [Epub ahead of print as doi:10.1210/jc.2011-1995.]. (2) Xu J et al., National Center for Health Statistics 2010; May 20, 58(19). (3) World Health Organisation Mortality Statistics. http://data.euro.who.int/dmdb/Help/inds.htm. Accessed: 22 March 2011.[br][br]Sources of Research Support: Funded by Eli Lilly and Company.[br][br]Disclosures: HJ: Employee, Eli Lilly & Company. AGZ: Employee, Eli Lilly & Company. DM: Employee, Eli Lilly & Company. LLR: Scientific Board Member, Eli Lilly & Company; Speaker, Eli Lilly & Company. E-ME: Scientific Board Member, Eli Lilly & Company; Speaker, Eli Lilly & Company. SWJL: Scientific Board Member, Eli Lilly & Company; Speaker, Eli Lilly & Company. SM: Scientific Board Member, Eli Lilly & Company; Speaker, Eli Lilly & Company. AFA: Scientific Board Member, Eli Lilly & Company; Speaker, Eli Lilly & Company; Consultant, Eli Lilly & Company. 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 551 74 777 SAT-699 PO21-01 Saturday 716 2012


717 ENDO12L_SAT-700 POSTER SESSION: GH Deficiency: Diagnosis, Prognosis [amp] Therapy (1:30 PM-3:30 PM) Sustained-Release Recombinant Human GH Improved Quality of Life in Adults with GH Deficiency Youngsook Kim, Jae Won Hong, Young Duk Song, Eun Jig Lee Yonsei University College of Medicine, Seoul, Korea; NHIC Ilsan Hospital, Ilsan, Gyeonggi-do, Korea [bold]Background:[/bold] Administration of recombinant human GH (rhGH) in adults with GH deficiency has been known to improve their metabolic impairment and Quality of life in previous studies. However, they suffer inconvenience from daily injection of rhGH.[br][bold]Objectives:[/bold] To evaluate the effects, safety, and compliance of weekly administered sustained-release recombinant human GH (SR-rhGH) supplement in patients with GH deficiency.[br][bold]Design:[/bold] This is a 12-week prospective, single-arm, open-label trial.[br][bold]Intervention/Participants:[/bold] Men and women aged [ge] 20 years with documented acquired GH deficiency (caused by pituitary tumor, trauma, other pituitary disease) are eligible for this study. All subjects were given 2 mg (6 IU) of SR-rhGH, administered subcutaneously, weekly for 12 weeks. Efficacy and side effect were assessed at baseline and within 30 days after 12[sup]th[/sup] injection. Comparison between baseline values and those at end time was performed.[br][bold]Measurements:[/bold] We evaluated AGHDA score for quality of life and measured serum IGF-1 level.[br][bold]Results: [/bold]The mean baseline IGF-1 level of 108.67 [plusmn] 73.03 ng/ml increased to 129.01 [plusmn] 68.37 ng/ml (P=0.0111) at week 12 and the mean baseline (AGHDA score) decreased 9.80 [plusmn] 6.51 to 7.55 [plusmn] 5.76 (P[lt]0.0001) at week 12. Adverse events related medication included pain, edema, rash, and heating sense at the administration site and these disorders diminished during investigation.[br][bold]Conclusions: [/bold]Weekly administered SH-rhGH for 12 weeks increased IGH-1 level effectively and improved quality of life in patients with GH deficiency without severe adverse events.[br][br]Nothing to Disclose: YK, JWH, YDS, EJL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 347 74 778 SAT-700 PO21-01 Saturday 717 2012


718 ENDO12L_SAT-701 POSTER SESSION: GH Deficiency: Diagnosis, Prognosis [amp] Therapy (1:30 PM-3:30 PM) Comparison of Intuitiveness, Ease of Use and Preference in Three Disposable Growth Hormone Injection Devices Anne-Marie Kappelgaard, Marianne Qvist, Franziska Winter, Britta Muller Novo Nordisk A/S, S[oslash]borg, Denmark; Novo Nordisk A/S, S[oslash]borg, Denmark; ikfe CRO GmbH, Mainz, Germany [bold]AIM: [/bold]Growth hormone (GH) is used to treat short stature in children with a variety of conditions including growth hormone deficiency (GHD), Turner syndrome (TS) and born small for gestational age (SGA). The need for daily injections can compromise adherence. Thus it is important that GH injection devices are convenient, practical and user-friendly. We compared the intuitiveness and ease of use of Norditropin[reg] FlexPro[reg] (FP, Novo Nordisk A/S, Bagsvaerd, Denmark) with two disposable devices, Nutropin AQ[reg] NuSpin[sup]TM[/sup] pen (NS, Genentech, USA) [Study INT1] and Genotropin[reg] GoQuick[reg] pen (GQ, Pfizer Inc, New York, USA) [Study INT2].[br][bold]METHODS: [/bold]In two randomised, non-interventional crossover studies, children with GHD, TS or SGA (aged 10[ndash]17 years, treated with GH for [ge]6 months) were randomly assigned to intuitiveness (INT1; INT2) (n=32; n=32) or instruction (n=32; n=32) groups. All patients participated in a usability test involving dose setting and injection into an Eppendorf tube. Time to inject and application errors were recorded. Intuitiveness groups had brief verbal instruction on device use. Instructed groups were instructed according to the user guide. Patient preference for the devices, intuitiveness and ease of use were assessed by questionnaire.[br][bold]RESULTS: [/bold]Mean [SD] injection times were shortest with FP in both intuitiveness (INT1 [FP vs. NS]/INT2 [FP vs. GQ]) (37.1 [10.6] vs. 81.5 [68.4], [italic]p[/italic][lt]0.001/39.8 [17.0] vs. 65.6 [42.9], [italic]p[/italic][lt]0.01) and instruction groups (32.2 [8.8] vs. 40.4 [14.5] [italic]p[/italic][lt]0.001/40.7 [19.7] vs. 48.1 [25.8], [italic]p[/italic][lt]0.05). Fewer errors were made by patients using FP, except in INT1 where errors (n) were similar for FP and NS (2 vs. 1) in the instruction group. In the device preference questionnaire (13 items), FP was the preferred device in the majority of questions in both intuitiveness (INT1 and INT2) (11/13 and 10/13) and instructed groups (11/13 and 10/13). In both studies, FP was chosen by most patients as their overall preference in the intuitiveness (INT1; INT2) (FP 19 vs. NS 7; FP 19 vs. GQ 4) and instructed groups (FP 23 vs. NS 4; FP 18 vs. GQ 5). More patients in the intuitiveness groups were confident in using FP without instruction (INT1: 97%; INT2: 97%) compared with NS (44%) or GQ (31%).[br][bold]CONCLUSIONS: [/bold]Both instructed and uninstructed patients preferred Norditropin[reg] FlexPro[reg] to comparator devices. Norditropin[reg] FlexPro[reg] required less time for injection, was associated with fewer application errors, and was rated as more intuitive to use.[br][br]Sources of Research Support: Novo Nordisk.[br][br]Disclosures: A-MK: Employee, Novo Nordisk; Stockholder, Novo Nordisk. MQ: Employee, Novo Nordisk. Nothing to Disclose: FW, BM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 732 74 779 SAT-701 PO21-01 Saturday 718 2012


719 ENDO12L_SAT-702 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) A Rare Case of Hereditary Syndrome of Inappropriate Anti-Diuretic Hormone (SIADH) Ranganatha Krishna Rao, Narendra Reddy, Narasimha Murthy, Martin O Weickert, Harpal Randeva, Sailesh Sankar University Hospital, Coventry, UK Introduction:[br]Hereditary SIADH or Nephrogenic syndrome of inappropriate anti-diuresis (NSIAD) is an X-linked disorder due to gain-of-function mutation of vasopressin receptor AVPR2, resulting in hyponatraemia and oliguria (1). We present a case of this rare hereditary condition affecting water and electrolyte balance.[br]Case report:[br]A 57-year old male with learning disabilities was referred by the renal physicians for persistent hyponatraemia evaluation. He had multiple admissions related to hyponatraemia and seizures. Given his learning disability, it was difficult to control his fluid intake. He did not have polyuria. He was clinically euvolaemic and systemic examination was unremarkable.[br]He had previously treated pulmonary tuberculosis and deep vein thrombosis. He was not on any medication.[br]Results:[br]Laboratory evaluation revealed: Plasma sodium 125 mmol/L (135-145), plasma osmolarity 260 mmol/kg (275-295), urine osmolarity 554 mmol/kg, urine sodium 89 mmol/L([lt]40), plasma urea 5 mmol/L (2.5-7.8), plasma creatinine 91 [micro]mol (70- 120). TSH 1.74 mU/L (0.5-6), random cortisol 832 nmol/L ([gt]550). Anti-diuretic hormone level was 0.9 pmol/L (unsuppressed) with corresponding plasma sodium 123 mmol/L, plasma urea 5.1 mmol/L, plasma osmolarity 252 mmol/kg. Brain magnetic resonance imaging was normal. Plasma sodium did not normalise (ranged from 116 to 132) despite fluid restriction (1litre/24 hours) and demeclocycline (600 mg/day). A diagnosis of hereditary SIADH was suspected, given the chronic nature and relatively asymptomatic status. Furthermore, his carer reported that his grand niece (aged 18 months) was admitted with hyponatremia. Further tests revealed hemizygous pathogenic missense mutation in exon 2: c.409 C[gt]T (p.Arg137Cys) in the AVPR2 gene by DNA sequence analysis, consistent with hereditary SIADH.[br]Progress[br]Usually patients avoid excess fluid by inherent nature of the condition as this makes them feel unwell. Although he has been advised on fluid restriction, given his learning difficulties, this has proved to be difficult. 5 of his family members were subsequently diagnosed to have the same mutation (sister, nephew, 2 nieces and grandniece).[br]Conclusion[br][bull] Although rare, nephrogenic SIADH should be considered in hyponatraemia patients especially when a family history of hyponatremia is present.[br][bull] Patients and relatives should be educated about recognition of signs and symptoms of hyponatraemia, careful fluid balance techniques and the need for family screening.[br][br]1. Decaux G, Vandergheynst F, Bouko Y, Parma J, Vassart G and Vilain C: Nephrogenic syndrome of inappropriate antidiuresis in adults: high phenotypic variability in men and women from a large pedigree. Journal of the American Society of Nephrology: JASN 18: 606-612, 2007.[br][br]Nothing to Disclose: RKR, NR, NM, MOW, HR, SS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2213 75 780 SAT-702 PO55-02 Saturday 719 2012


720 ENDO12L_SAT-703 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) The Unusual Development of Drug-Induced SIADH Immediately after Transsphenoidal Pituitary Surgery (TSS) Christina M Trillis, Baha Arafah University Hospitals Case Medical Center, Cleveland, OH Background:[br]Fluid and electrolyte disturbances are a well known complication following TSS with diabetes insipidus (DI) being the most common manifestation in the first 24 hours. SIADH has been reported less commonly with a peak incidence between days 6 to 9 after surgery. The cause of SIADH in the latter setting is likely from ADH leakage from damaged neurohypophyseal axons. Other causes of SIADH are not commonly reported during this time period. We herein report a patient who developed Valproic Acid induced SIADH on the third day following TSS.[br]Clinical Course:[br]A 66 year old woman with a hemorrhagic Rathke[apos]s cleft cyst (Rathke[apos]s cleft cyst apoplexy) presented with central hypogonadism, hypothyroidism, and growth hormone deficiency, and was admitted for TSS. Before, and for the first 48 hours after the surgery, she had documented normal posterior pituitary function. On the second post-op day she developed a severe migraine which she has had previously, and which could not be controlled with large doses of narcotics. CT of the head was negative. One dose of Valproic Acid (1000 mg IV) was administered 80 hours post-op to control the migraine attack. Five hours later, she developed severe confusion and agitation. At that time, she had normal vital signs and was euvolemic. However lab studies showed a serum Na of 116 mmol/L, a K of 3.1 mmol/L, a creatinine of 0.76 mg/dL, a free T4 of 0.83 ng/dL, and a serum glucose of 124 mg/dL. Simultaneous serum and urine osmolality were 242 and 526 mOsm/kg H2O, respectively. Simultaneous measurement of plasma ADH revealed a level of 3.8 pg/mL, which is clearly inappropriately elevated considering the degree of serum hypoosmolality. A random serum cortisol was 12.7 ug/dL with a serum prolactin level of 6.8 ug/L. These data indicated intact anterior pituitary hormone secretion. The patient was treated in the ICU with 3% NS and fluid restriction. The sodium normalized over a 48 hour span. Subsequently she developed a hematoma at the surgical site that created stalk mass effect causing hypopituitarism with elevated serum prolactin level. She was given hydrocortisone for a week, following which it was withdrawn. She has continued to do well a year after her surgery with documented normal posterior pituitary function, as well as normal pituitary-thyroidal and adrenal functions.[br]Conclusion: If symptoms of SIADH occur outside of the usual timeline following TSS, other etiologies must also be appropriately investigated.[br][br]Cusick JF, et al. In appropriate secretion of antidiuretic hormone after transsphenoidal surgery for pituitary tumors. N Engl J Med. 1984 Jul 5; 311(1) 36-8. Beers E, et al. Syndrome of inappropriate antidiuretic hormone secretion (SIADH) or hyponatremia associated with valproic acid: four case reports from the Netherlands and a case/non-case analysis of vigibase. Drug Saf. 2010 Jan 1; 33(1): 47-55. Chaiban, J et al. Rathke cleft cyst apoplexy: a newly characterized distinct clinical entity. J Neurosurg. 2011 Feb;114(2):318-24.[br][br]Nothing to Disclose: CMT, BA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 694 75 781 SAT-703 PO55-02 Saturday 720 2012


721 ENDO12L_SAT-704 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) A Case Report of Severe Hypotonic Hyponatremia Secondary to Concomitant Intake of Desmopressin Replacement Therapy and Non-Steroidal Anti-Inflammatory Drugs Emanuele Ferrante, Elisa Verrua, Giovanna Mantovani, Andrea Noto, Elisa Sala, Elena Malchiodi, Gaetano Iapichino, Paolo Beck-Peccoz, Anna Spada Endocrinology and Diabetology Unit, Fondazione IRCCS C[agrave] Granda Ospedale Maggiore Policlinico, Milan, Italy; Anesthesia and Intensive Care Unit, Milan, Italy; Intensive Care and Dermatological Sciences, Ospedale San Paolo, Milan, Italy Background: Desmopressin (DDAVP) is an effective treatment of both polyuric conditions and bleeding disorders. Most of the data on safety profile of desmopressin (DDAVP) derives from studies on titration of drug treatment, whereas few reports describe severe side effects secondary to drug-drug interaction.[br]Clinical case: We report the case of a 50-yr-old Caucasian man, affected with congenital central diabetes insipidus (DI), admitted to the Emergency Unit for ingravescent nausea, emesis and weakness since twelve hours. A recent endocrinological evaluation reported a normal anterior pituitary function. Shortly after admission, he developed an episode of generalized tonic-clonic seizure, resulting in coma. The post-critic physical examination did not showed clinical signs of volume expansion or depletion. Moreover, brain CT scan was normal. Based on his medical history and clinical findings, water intoxication secondary to non steroidal anti-inflammatory drug (NSAID) intake (ketoprofen, 200 mg/day for the last 3 days for cervical pain) concomitant with DDAVP replacement therapy (Minirin[reg] 60 mcg 4 tablets a day) was hypothesized as cause of the severe euvolemic hypotonic hyponatremia observed at entry (natremia 113 mEq/l, n 135-145 mEq/l; plasma osmolality 238 mOsm/Kg, n 275-295 mOsm/Kg) and the acute fall in sodium levels was considered responsible for seizures and coma. After emergency procedures, the aquaretic tolvaptan (Samsca[reg] 7.5 mg) was administered and hydratation was maintained according to water excretion. At the same time, desmopressin substitution was withdrawn. The patient completely recovered in 72 hours and was discharged 10 days after with reinstatement of desmopressin replacement therapy.[br]Conclusions: Although several cases of hyponatremia in patients on DDAVP for different indications have been reported, no study has so far highlighted the potentially life-threatening side effects associated with NSAIDs intake during DDAVP replacement therapy for central diabetes insipidus. In order to improve DDAVP safety, risks and benefits of co-treatment should be carefully considered and therapeutic alternatives to NSAIDs should be recommended to patients with central diabetes insipidus.[br][br]Nothing to Disclose: EF, EV, GM, AN, ES, EM, GI, PB-P, AS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1439 75 782 SAT-704 PO55-02 Saturday 721 2012


722 ENDO12L_SAT-705 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Diabetes Insipidus in Pregnancy Complicated by Pre-Eclampsia and Sickle Cell Trait Rajeev Sharma, Mary Toussaint-Milord, Agnieszka Gliwa, MaryAnn Banerji State University of New York (SUNY) Downstate Medical Center, Brooklyn, NY; State University of New York (SUNY) Downstate Medical Center, Brooklyn, NY Introduction: Diabetes insipidus (DI) in pregnancy is a rare disorder complicating about 1 in 30000 pregnancies. Excessive vasopressinase activity (a placental enzyme which degrades arginine vasopressin) is thought to be the underlying pathology. We report a rare case of DI in pregnancy associated with pre-eclampsia and sickle cell trait.[br]Clinical Case: A 19-year-old black female at 34 weeks gestation was admitted with symptoms of excessive thirst, urination, and headache for 6-8 weeks. She was not on any medications. BP was 160/98 mmHg; otherwise physical examination was normal. 24-hour urine output was 11.6 L with increased urinary proteins (928 mg/24 hrs). Urine specific gravity was 1.002 (2 months ago it was 1.011). Serum sodium was 139 mEq/L; other electrolytes were normal. Liver enzymes were mildly elevated (AST 72, ALT 29) Serum osmolality was elevated for pregnancy (286 mOsm/kg) with low urine osmolality (103 mOsm/kg). Blood arginine vasopressin levels were low normal (0.8, n= 0-4.7 pg/mL). Anterior pituitary hormonal work up and blood glucose levels were normal. She was given intravenous normal saline, MgSO4, and 4 mcg of dDAVP subcutaneously. Urine output decreased with increase in osmolality to 431 mOsm/kg. The patient underwent an uncomplicated C-section and 24 hours later the urine osmolality decreased again with increasing urine output. Interestingly, urine osmolality never reached more than 500-600 mOsm/kg prompting us to seek another underlying pathology. On further questioning, patient admitted having sickle cell trait, which was confirmed with hemoglobin electrophoresis. She also reported nocturia (1-2 times) since childhood. A literature search revealed that sickle cell trait could mimic nephrogenic DI. Patient was started on oral dDAVP 0.1 mg twice a day and discharged with outpatient follow up for further management including formal water deprivation test and possible imaging of pituitary.[br]Conclusion: Diabetes insipidus in pregnancy with pre-eclampsia is a rare but well-known entity. However it may be complicated by other underlying conditions. Sickle cell trait/disease can affect the renal medulla causing impaired concentration ability and decreased urinary osmolality and polyuria. Interestingly, subclinical DI may be unmasked in pregnancy as reported in literature and in this case. (Iwasaki et al., NEJM 1991; 324: 522) Therefore, it is important to consider other differentials when evaluating a case of polyuria in pregnancy.[br][br]Iwasaki et al., NEJM 1991;324:522.[br][br]Nothing to Disclose: RS, MT-M, AG, MB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1355 75 783 SAT-705 PO55-02 Saturday 722 2012


723 ENDO12L_SAT-706 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Early-Onset of Diabetes Insipidus Following Brachytherapy for Recurrent Pituitary Adenoma Hien T Tran, Roe Erin, Robert Timmerman, Hatanpaa Kimmo, Ildiko Lingvay University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX [underline]Case[/underline]:[br]A 26 year-old Hispanic man with history of a pituitary macroadenoma presented with headache and visual field defects to our hospital seven years after undergoing two transcranial and one transphenoidal resection of a 3.2x1.1x3.4cm adenoma. Pathology from the final resection at the other facility stained positive for ACTH. He was then lost to follow-up for 3 years. At our hospital, he was found to have bulky tumor regrowth encroaching on the optic chiasm. Given the tumor size, character, and multiple recurrences, a multi-disciplinary group did not believe that another surgery was the best option. Given the proximity to the optic chiasm, rather than external radiotherapy, interstitial radiation with [sup]125[/sup]I was performed to help reduce mass effect.[br][underline]Results:[/underline][br]Two [sup]125[/sup]I seeds were stereotactically placed into the pituitary tumor via a transnasal/transphenoidal approach. Prior to seed implantation, the patient had signs and symptoms of hyperglycemia and weight gain, but no other evidence of hypercortisolism. At the time of seed implantation, his pituitary function was intact aside from having hypogonadotropic hypogonadism. Six weeks after seed implantation, a CT head confirmed a dramatic reduction of the suprasellar and pre-pontine portions of the adenoma resulting in improvement in vision. At that time, he was found to have central hypothyroidism. The patient[apos]s hyperglycemia resolved two months after [sup]125[/sup]I implantation with concomitant sixty pounds weight loss. Six months after seed implantation, the patient presented with polyuria and polydipsia not related to glycemic control. At that time, he was found to have new-onset central diabetes insipidus by DDAVP provocation. He was also found to have new onset of adrenal insufficiency.[br][underline]Conclusions: [/underline][br]Use of high activity [sup]125[/sup]I interstitial implants for a large and invasive pituitary adenoma in our patient resulted in a dramatic tumor response with improvement in vision. This degree of tumor response would be unusually after conventional external radiotherapy. However, he also experienced decline/loss of both anterior and posterior pituitary function in a timeframe considerably earlier than expected after external beam irradiation. This treatment warrants further investigation.[br][br]Nothing to Disclose: HTT, RE, RT, HK, IL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2044 75 784 SAT-706 PO55-02 Saturday 723 2012


724 ENDO12L_SAT-707 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Endocrine Involvement in Langerhans Cell Histiocytosis Joana Couto, Raquel G Martins, Ana P Santos, Isabel Torres Portuguese Institute of Oncology FG of Porto, Porto, Portugal Background: Langerhans cell histiocytosis (LCH) is a rare granulomatous disease of unknown etiology, that may affect a variety of organs and often involves the hypothalamo-pituitary axis (HPA). Central Diabetes insipidus (CDI) is the most frequent endocrine abnormality, developing in up to 25% of cases. Patients (pts) with multisystem disease and craniofacial involvement at the time of diagnosis have an increased risk of developing CDI. Anterior pituitary dysfunction is found more rarely and is usually associated with DI.[br]Clinical cases: We retrospectively reviewed data from five pts with histopathological diagnosis of LCH followed at the Endocrinology Department of our Institution. Three female and 2 male pts were identified. The mean age at diagnosis was 4 years (range: 3-6). They were followed for a median of 15.5 years (range:1.5-29). Four patients had skull and/or maxillofacial bone involvement; one pt had lymph node involvement. All of them presented evidence suggestive of hypothalamo-pituitary involvement, namely CDI. In three patients, CDI was the presenting manifestation of LCH. In two pts CDI was verified, one and four years after LCH diagnosis. One patient presented with CDI one year before LCH diagnosis was established. None of the five pts developed, so far, anterior pituitary hormonal deficiencies. Radiological examination of the HPA was abnormal in one pt. All 5 pts were treated with chemotherapy and 3 patients received external beam radiotherapy.[br]Conclusion: These observations confirm that CDI is the main endocrine manifestation of LCH, being often the presenting feature of the disease. So, the clinician should be alert for symptoms and signs suggesting endocrine involvement in all LCH pts. In pts presenting with apparent idiopathic CDI, LCH ought to be considered and possible extracranial locations should be evaluated.[br][br]Nothing to Disclose: JC, RGM, APS, IT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 592 75 785 SAT-707 PO55-02 Saturday 724 2012


725 ENDO12L_SAT-708 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Hypothalamic-Pituitary Involvement and Bone Lesions in a Patient with Erdheim-Chester Disease: The Effect of 12 Months of Zoledronic Acid Treatment Fatih Tanriverdi, Zuleyha Karaca, Ahmet Selcuklu, Ozlem Canoz, Gulsah Elbuken, Halil Donmez, Ummuhan Abdulrezzak, Kursad Unluhizarci, Fahrettin Kelestimur Erciyes University Medical School, Kayseri, Turkey; Erciyes University Medical School, Kayseri, Turkey; Erciyes University Medical School, Kayseri, Turkey; Erciyes University Medical School, Kayseri, Turkey; Erciyes University Medical School, Kayseri, Turkey [bold]Background:[/bold] Erdheim-Chester disease (ECD) is a very rare xanthogranulomatous non-Langerhans cell histiocytosis of unknown origin, caused by infiltration of foamy lipid-laden macrophages. It is characterized by bilateral symmetrical medullary sclerosis at the diametaphyseal portion of long bones of the lower extremity. Extraskeletal involvement mainly includes heart, lung, and kidneys. In the literature only 5 cases with biopsy proven ECD were defined with the involvement of hypothalamic-pituitary area. Our patient was the first case initially presented with diabetes insipidus (DI) then progress to panhypopituitarism and treated with 12 months of zoledronic acid.[br][bold]Clinical case: [/bold]A 57 year old man was admitted with the complains of polyuria and polydipsia in 1997 and diagnosed as DI. On his initial MRI scan pituitary stalk was thickened and posterior signal intensity was lost. Based on the clinical data he was diagnosed as lymphocytic hypophysitis and desmopressin was given. Throughout 3 years follow-up he developed panhypopituitarism and was commenced hormonal replacement. Despite hormone replacement his fatigue, headache and generalized bone pain were worsened. When MRI images in 2001 and 2009 were compared a nodular lesion was formed on the thickened stalk and this lesion progressively reached to suprasellar area. He was operated on transsphenoidally in 2009 and based on the histological and immunohistochemical findings he was given a diagnosis of non-langerhans cell histiocytosis. After this pathological finding we re-evaluated the patient with bone X-rays and bone scintigraphy. On x-ray typical bilateral sclerotic lesions were present on both metaphysis of tibia and femur. Bone scintigraphy revealed multiple bone involvement including long bones of the lower extremities, skull, mandibular and maxillary bones. After biochemical and radiological evaluation no other organ involvement was defined. Initially serum pyridinoline level was slightly higher than the normal reference range. We started zoledronic acid treatment and continued for 12 months. During follow-up the patient was clinically stable and he had no more bone pain. Although the lesions on bone scintigraphy did not disappear the intensities of the uptake were slightly decreased and the lesions did not progress.[br][bold]Conclusion:[/bold] In the present case zoledronic acid has been found as effective, at least stabilized the lesions, in the treatment of the ECD without any important side effect.[br][br]Nothing to Disclose: FT, ZK, AS, OC, GE, HD, UA, KU, FK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 189 75 786 SAT-708 PO55-02 Saturday 725 2012


726 ENDO12L_SAT-709 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Lymphocytic Pan-Hypophysitis as a First Presentation of Sj[ouml]gren Syndrome [mdash] Our Experience with a Case Cherng Jye Seow, Rinkoo Dalan Tan Tock Seng Hospital, Singapore, Singapore [bold]Background and Introduction[/bold]: Sjogrens syndrome (SS) has been associated with autoimmune hypophysitis in only 0.8% of all cases (1). We present a case who presented with cranial diabetes insipidus (CDI) from lymphocytic panhypophysitis (LPH) secondary to SS.[br][bold]Case Presentation[/bold]: A 61-year-old gentleman presented with progressive generalised weakness, unsteadiness, loss of weight over 6 months and recent polyuria (5.3 litres urine/day). Biochemistry: Serum sodium 162 mmol/l, serum osmolality 313 mosm/L, Urine Osmolality [lt]130mmol/L suggestive of diabetes insipidus. A test dose of DDAVP resulted in urine osmolality concentrating 4-fold, establishing the diagnosis of CDI. He also had low IGF-1 levels and hypogonadotrophic hypogonadism. MRI brain: multiple posterior fossa enhancing lesions and a thickened and enhanced pituitary stalk. Further investigations revealed a positive ANA titre (320units/ml) and anti-Ro titre 197 units/ml (RI: [lt]20). Retrospectively, he did have long standing sicca symptoms of dry eyes and mouth. Schirmer[apos]s test was positive with 0mm tear flow and a salivary gland biopsy showed lymphocytic sialoadenitis thus suggestive of SS. He was started on DDAVP, high dose prednisolone and immunosupressants (cyclophosphamide/hydroxychloroquine). A subsequent MRI in 3 months showed interval resolution of the stalk thickening suggestive of a regression.[br][bold]Discussion[/bold]: In LPH, the autoimmune process either simultaneously targets both the pituitary lobes, or the process is initially confined to one lobe which spreads to the other. Initially, the pituitary is inflamed, infiltrated by lymphocytes and at this stage patients may have subclinical endocrine disease. If the inflammation resolves, either spontaneously or with the aid of glucocorticoids, and the pituitary parenchyma is not destroyed, remission occurs. If inflammation progresses, the pituitary is replaced by fibrotic tissue and becomes atrophic and loses its function (1). Although the corticotrophes and thyrotrophes are more commonly involved in LPH, GH and gonadotrophes may be involved early as well like in our case.[br][bold]Conclusion[/bold]: LPH as a presenting symptom for SS is extremely rare (1,2), and a high index of suspicion is necessary to make the diagnosis. Early treatment results in increased chances of remission.[br][br]1) Caturegli et al. Autoimmune hypophysitis. Endocrine reviews 2005; 26(5):599-614. 2) JY Li et al. Hypertrophic cranial pachymeningitis and lymphocytic hypophysitis in Sjogren[apos]s syndrome. Neurology 1999. Jan 15;(52):420-3.[br][br]Sources of Research Support: [bold]Nothing to Declare[/bold]: SCJ, DR[br][br]Nothing to Disclose: CJS, RD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 96 75 787 SAT-709 PO55-02 Saturday 726 2012


727 ENDO12L_SAT-710 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Ipilimumab-Associated Hypophysitis [mdash] Time Course of MRI and Hormonal Changes Vijay G Eranki, Georges Elhomsy, Alan Silverberg, Stewart Albert Saint Louis University School of Medicine, St Louis, MO Introduction[br]Ipilimumab (Yervoy), a monoclonal antibody against cytotoxic T-lymphocyte antigen-4, is approved for chemotherapy of metastatic melanoma. This antibody augments T-cell activation, proliferation and anti-tumor immunity. Adverse effects include immune-mediated endocrinopathies. We report the natural history of Ipilimumab associated hypophysitis.[br]Clinical Case[br]A 76 year old man with melanoma diagnosed three years previously had undergone surgical resection, radiation and chemotherapy. With tumor recurrence in the lungs, Ipilimumab was instituted at 3 mg/kg every three weeks for four doses. Four weeks after completing the regimen, he presented with severe fatigue, headache and blurry vision. Blood pressure was 127/79 without orthostatic changes. There was no hyper-pigmentation. Visual fields and ophthalmological funduscopic examinations were normal.[br]Laboratory tests: TSH [lt]0.015 [mu]IU/mL (0.35-5.5), free T4 [lt]0.4 ng/dL (0.7-1.9), ACTH 2.5 pg/mL (7.2-63), and diminished response to cosyntropin stimulation test (CST) [baseline serum cortisol (Ct0) [lt]0.2 [mu]g/dL (4.3-28) and 1-hour(Ct1) 6.8 [mu]g/dL]. MRI showed a hyper-enhancing pituitary gland (volume = 900 mm3) which was distinctly different from the pre-therapy MRI, which was non-enhancing (volume = 354 mm3). The patient was diagnosed with hypophysitis and treated with prednisone (5mg/day) and levothyroxine. Five months later, there was resolution of the hyper-intense pituitary uptake (pituitary volume = 115 mm3). Repeat endocrine evaluation was: TSH = 0.19 [mu]IU/mL, ACTH [lt]1.1pg/mL and CST with (Ct0) = 0.2[mu]g/dL, (Ct1) = 3.0 [mu]g/dL.[br]Discussion/Conclusion[br]The patient presented with symptoms which may have been ascribed to tumor cachexia. Endocrine and neuro imaging evaluation confirmed hypophysitis. Five months after discontinuation of the drug, there was a reversal of the hypophysitis on MRI but not in recovery of ACTH. Although, it is suggested that the hypophysitis be treated with high dose steroids, the patient received a physiological replacement dose in an attempt not to reverse the immunological anticancer effects. Ipilimumab induced hypophysitis should be evaluated with appropriate hormonal and MR imaging in a patient with [apos]tumor cachexia[apos].[br][br]Min L, Vaidya A, Becker C.Ipilimumab therapy for advanced melanoma is associated with secondary adrenal insufficiency: a case series. Endocrine Practice 2011.[br][br]Nothing to Disclose: VGE, GE, AS, SA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 907 75 788 SAT-710 PO55-02 Saturday 727 2012


728 ENDO12L_SAT-711 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Central Adrenal Insufficiency and Hypothyroidism after Ipilimumab Treatment Sunna Kwun, Ivana Lukacova-Zib, Geetha Gopalakrishnan Alpert Medical School of Brown University, East Providence, RI [bold]Introduction:[/bold] Ipilimumab is a targeted antibody that activates a T cell mediated immune response against tumor cells or other antigens. It is used in the treatment of various cancers including metastatic melanoma. Immune reactions to self antigens result in a wide array of adverse events including enterocolitis, hepatitis and endocrinopathies. Although Ipilimumab induced endocrinopathies are rare, the consequences can be severe and typically occur 10 weeks after treatment initiation. We report a case of Ipilimumab induced autoimmune hypophysitis presenting 4 weeks after initial therapy.[br][bold]Clinical Case[/bold]: A 48 year old female with metastatic melanoma presents with severe headache approximately 4 weeks after initiating Ipilimumab. MRI identified an enlarged pituitary gland measuring 1.2 cm in the craniocaudal dimension. Prior imaging was notable for a normal pituitary gland measuring 5.4 mm. Pituitary enhancement was suspicious for lymphocytic hypophysitis or metastatic disease. The patient was continued on Ipilimumab and placed on high dose steroids to treat the autoimmune process. Evaluation of pituitary function was notable for a TSH 0.3 (0.35-5.50 uIU/ml), FT4 0.73(0.8-1.8ng/d.), Cortisol AM 0.7 (6.2 to 19.4ug/dl), Prolactin 1(3-29ng/ml) and IGF-1 85 (118-298ng/ml) suggesting hypopituitarism. The patient was treated with thyroid hormone replacement and the steroid dose was tapered to maintenance levels upon resolution of headache symptoms. Subsequent imaging showed a stable, non-FDG avid pituitary lesion on PET-CT.[br][bold]Conclusion:[/bold] The autoimmune effects of Ipilimumab may lead to hypophysitis and other endocrinopathies. Since adverse events can occur early in the treatment course, we recommend clinical evaluations to focus on the pituitary, adrenal and thyroid axis after each treatment cycle and with symptoms. Laboratory and brain imaging can be considered based on clinical assessments.[br][br]1.Blansfield, Joseph et al. Cytotoxic T-Lymphocyte-Associated Antigen-4 Blockage Can Induce Autoimmune Hypophysitis in Patients with Metastatic Melanoma and Renal Cancer. J Immunother 2005; 28:593-598. 2.Dillard, Troy et al. Anti-CTLA-4 antibody therapy associated autoimmune hypophysitis: serious immune related adverse events across a spectrum of cancer subtypes. Pituitary 2010; 13:29-38. 3. Yang, James et al. Ipilimumab (anti-CTLA4 Antibody) Causes Regression of Metastatic Renal cell Cancer Associated with Enteritis and Hypophysitis. J Immunother 2007;30:825-830.[br][br]Nothing to Disclose: SK, IL-Z, GG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2384 75 789 SAT-711 PO55-02 Saturday 728 2012


729 ENDO12L_SAT-712 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Interferon-Induced Hypopituitarism Presenting as Hypothyroid Myopathy Johnson Thomas, Jignesh Shah, Monika Datt, Kenneth H Hupart NuHealth, Nassau University Medical Center, East Meadow, NY; St John[apos]s University, Queens, NY; Albert Einstein College of Medicine, Bronx, NY Interferon (IFN), particularly when used to treat Hepatitis C, has been associated with the development of autoimmune endocrinopathies. IFN induced thyroid failure is widely recognized but pituitary dysfunction secondary to IFN is rare. We report a patient who presented with selective hypopituitarism 4 months after completing a course of IFN with symptoms that did not initially lead to a timely diagnosis.[br]A 60 yo female was admitted with body aches, muscle cramps, and weakness for 3 mos. She also experienced a change in voice, dysphagia, intermittent hand numbness and facial edema during this time. Four months ago, she completed a 6 month course of IFN [alpha][ndash]2a for Hepatitis C. Past history: Type 2 DM treated with basal insulin and a sulfonylurea. PEx revealed BP-165/76, HR-73, T-97.7[ordm]F. Facial puffiness and lingual swelling were present. Thyroid gland was low, no Pemberton[apos]s sign. Kussmaul[apos]s sign was absent. Heart, breath and bowel sounds were normal. Neurological exam revealed proximal muscle weakness and delayed DTR relaxation. Initial lab tests: Na=126 mmol/L, Glucose=217 mg/dL, CPK=6761 U/L, and TSH=2.26 mIU/L. Central hypothyroidism was considered leading to further testing: fT4=0.11 ng/dL, T4=0.79 ug/dL, Prolactin-undetectable, FSH=2.6 mIU/ml, LH=0.6 mIU/ml and IgF-1[lt]25 ng/ml. AM serum cortisol=11.7 ug/dL. Pituitary MRI was limited by movement artifact, but no mass was detected nor was pathologic signal enhancement observed. These findings lead to the diagnosis of anterior hypopituitarism with preservation of the adrenal axis; replacement therapy was initiated. The temporal relationship of this hypopituitarism to IFN therapy coupled with reports of IFN induced hypophysitis lead us to conclude that this was the likely underlying etiology of our patient[apos]s endocrine deficiencies.[br]Fatigue due to IFN is well known and often prompts an evaluation for primary hypothyroidism. Our patient was hospitalized with profound hypothyroidism, yet TSH was normal and the etiology of her symptoms was not identified until they became disabling. Few cases of hypophysisitis due to IFN have been reported; our patient lacked MRI features typically associated with autoimmune hypophysitis. The presence of central hypothyroidism [amp] deficiencies of GH, LH, FSH and prolactin points to major pituitary dysfunction that remained undiagnosed for months. With the frequent use of IFN, it is important to highlight the potential occurrence of this important adverse effect.[br][br]Nothing to Disclose: JT, JS, MD, KHH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2126 75 790 SAT-712 PO55-02 Saturday 729 2012


730 ENDO12L_SAT-713 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) A Case of Mixed Central and Nephrogenic Diabetes Insipidus Madiha Khan, Jasmine Boparai The University of Texas Medical Branch at Galveston, Galveston, TX [bold]Introduction:[/bold][br]Hydrocephalus is a rare cause of central diabetes insipidus. Lithium can lead to irreversible nephrogenic diabetes insipidus. A complicated case of diabetes insipidus with both central and nephrogenic components attributable to both of these causes is described below.[br][bold]Clinical Case:[/bold][br]A 62-year-old man was admitted to the hospital for altered mental status and was found to have hypernatremia. His past medical history included bipolar disorder, schizophrenia, hydrocephalus, stage 3 CKD and HTN. The patient was a resident of an assisted-living facility due to psychiatric illness. The patient had been on lithium for bipolar disorder in the past, which was stopped seven months prior to admission secondary to hypernatremia. Despite not taking Lithium for more than seven months, the patient complained of excessive thirst, polyuria and polydipsia at presentation. Physical exam revealed dehydration. Laboratory work up revealed a serum sodium of 163 mmol/L (normal, 135-145), plasma osmolality of 334 mosm/kg (normal, 278-305) and urine osmolality of 133 mosm/kg (normal, 50-1100). A CT scan of the head performed seven months prior to admission was reviewed and showed persistent communicating hydrocephalus. The patient was diagnosed with diabetes insipidus and intravenous hydration was started. Two hours after administration of Desmopressin 5 mcg subcutaneously, urine osmolality increased from 111 to 219 mosm/kg and urine output decreased from 220 cc/hr to 115 cc/hr. The patient was diagnosed with central diabetes insipidus secondary to almost 100% response to desmopressin. No significant abnormality of any other pituitary hormones was identified. The patient was started on Desmopressin treatment. Two months post-discharge, laboratory work-up showed a serum sodium of 140 mmol/L (normal, 134-146), plasma osmolality of 301 mosm/kg (n, 275-295) and urine osmolality of 289 mosm/kg(n, 300-800).The patient was able to concentrate urine but still not appropriately for the given plasma osmolality. The patient was diagnosed with mixed central and nephrogenic diabetes insipidus secondary to communicating hydrocephalus and history of lithium use.[br][bold]Conclusion:[/bold][br]Diabetes insipidus can result from mixed central and nephrogenic components in the presence of certain risk factors. A rare association of central diabetes insipidus with hydrocephalus should be considered.[br][br]Nothing to Disclose: MK, JB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1065 75 791 SAT-713 PO55-02 Saturday 730 2012


731 ENDO12L_SAT-714 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Diffuse Large B-Cell Lymphoma Presenting with Diabetes Insipidus: A Case Report Arshpreet Kaur, Asha Thomas Sinai Hospital of Baltimore, The Johns Hopkins-Sinai Hospital Internal Medicine Residency Program, Baltimore, MD; Sinai Hospital of Baltimore, Baltimore, MD [bold]Background:[/bold] The clinical presentation of B cell lymphoma as diabetes insipidus is rare. Pituitary involvement can occur either as primary CNS lymphoma or with metastatic disease (1). Involvement of the pituitary gland with lymphoma of paranasal sinuses has been rarely described (2).[br][bold]Clinical Case:[/bold] A 53 year old woman with a history of HIV presented with a six month history of polydipsia, polyuria, and intermittent headache. The patient reported a high volume urine output on an hourly basis. The physical examination was not revealing. Laboratory work revealed sodium of 145 mg/dl, low urine osmolality of 125 mOsm/kg, urine sodium of 25 mEq/L and no other pituitary hormone deficiency. MRI showed a 6mm lesion in the posterior pituitary and ill-defined decreased T1 signal in clivus and sphenoid sinus. She was treated medically with desmopressin for her urinary symptoms with some symptomatic improvement. The patient was referred for neurologic and neurosurgical evaluation. However, patient[apos]s headaches continued to worsen. In the interim, she was admitted for acute visual loss. Repeat imaging revealed a soft tissue mass involving the paranasal sinuses with intracranial extension to the sella and hypothalamus. Biopsy confirmed the diagnosis of diffuse large B cell lymphoma. Further imaging revealed cervical and inguinal adenopathy. Patient was treated urgently with multi-drug chemotherapy with dose adjusted EPOCH-R (etoposide, adriamycin, vincristine, cyclophosphamide, prednisone and rituximab) which resulted in neurological improvement. Intraventricular methotrexate was also added to the regimen. After 12 cycles, patient did well with regression of the tumor and marked improvement of her urinary symptoms with decreasing desmopressin requirement.[br][bold]Conclusion:[/bold] This is a rare case describing hypothalamic-pituitary involvement in diffuse large B cell lymphoma, due to paranasal sinus involvement with CNS extension, presenting with central diabetes insipidus (3,4). Early detection with a high index of suspicion, particularly in patients with HIV, and treatment can lead to better outcomes.[br][br](1) Kenchaiah M, Hyer SL. Diffuse large B-cell non Hodgkin[apos]s lymphoma in a 65-year-old woman presenting with hypopituitarism and recovering after chemotherapy: a case report. Journal of Medical Case Reports 2011; 5:498. (2) Tamer G, Kartal I and Aral F. Pituitary infiltration by non-Hodgkin[apos]s lymphoma: a case report. Journal of Medical Case Reports. 2009; 3:9293. (3) Megan OC, Payne S, Evanson J, Lister TA, Grossman AB. Lymphoma metastasizing to the pituitary: an unusual presentation of a treatable disease. Pituitary. 2005;8(2):139-46. Review. (4) Breidert M, Schimmelpfennig C, Kittner T, Helwig A, Ehninger G. Diabetes insipidus in a patient with a highly malignant B-cell lymphoma and stomatitis. Exp Clin Endocrinol Diabetes. 2000;108(1):54-8.[br][br]Nothing to Disclose: AK, AT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 707 75 792 SAT-714 PO55-02 Saturday 731 2012


732 ENDO12L_SAT-715 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Pituitary Pseudomacroadenoma in Severe Hypothyroidism: A Cautionary Tale Sonya Addison, David W Gardner University of Missouri, Columbia, MO [bold][underline]Background[/underline][/bold]: Pituitary hyperplasia commensurate with elevated serum TSH is known to occur in severe hypothyroidism, most notably in pediatric patients with severe primary hypothyroidism. The current case report attests to the same phenomenon in adults, and is offered as a reminder of the importance of comprehensive endocrine assessment prior to pituitary surgery.[br][bold][underline]Clinical case[/underline][/bold]: A 41-year-old women with a 15 year history of primary hypothyroidism presented with symptoms of severe fatigue, dry skin, constipation, heat intolerance, palpitations, headaches, and visual changes. Serum TSH was [gt] 150 mIU/L (normal [nl] 0.35-4.00 mIU/L), and MRI of the pituitary showed an enlarged, heterogeneously-enhancing pituitary, measuring 1.0 x 1.5 x 1.0 cm. The enlarged gland abutted but did not displace the optic chiasm. Neurosurgical intervention was contemplated but joint consultation with endocrinology resulted in resumption of levothyroxine maintenance therapy (the patient acknowledged chronic hypocompliance with her medication regimen). Five weeks later, TSH was 8.36 mIU/mL, and all symptoms were improved. Repeat MRI demonstrated shrinkage of the pituitary to 9 x 8 mm, and it no longer abutted the optic chiasm.[br][bold][underline]Discussion[/underline][/bold]: Pituitary hyperplasia secondary to severe hypothyroidism can resemble and actually be indistinguishable from a pituitary macroadenoma, and has at times resulted in unnecessary neurosurgical intervention. Thyrotroph hyperplasia in adults can also induce hyperprolactinemia, which can be confused with a prolactinoma. In either instance, thyroid hormone replacement therapy can lead to rapid resolution of thyrotroph hyperplasia and pituitary mass effects, as well as hyperprolactinemia. Levothyroxine offers far superior cost-effectiveness and lower risk than either transsphenoidal surgery or dopaminergic drug therapy.[br][bold][underline]Conclusion[/underline][/bold]: Evaluation of the pituitary-thyroid axis prior to surgical treatment is an important, but potentially overlooked, step in the workup of apparent pituitary macroadenoma. This simple step can prevent the risk and cost of a neurosurgical procedure, as well as possible long-term post-op morbidity, such as diabetes insipidus and hypopituitarism.[br][br]Nothing to Disclose: SA, DWG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 985 75 793 SAT-715 PO55-02 Saturday 732 2012


733 ENDO12L_SAT-716 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Acute Psychosis in a Patient with Prolonged Secondary Adrenal Insufficiency Following Initiation of Corticosteroids Eva S Liu, Carolyn Becker Brigham and Women[apos]s Hospital, Boston, MA [bold]Background:[/bold] Case reports have shown patients with Addison[apos]s disease can develop acute psychosis after initiation of corticosteroid therapy. The transient symptoms can arise even on physiologic replacement doses.[br][bold]Clinical Case:[/bold] A 48 year old male with history of asthma presented for transphenoidal resection of a 6.0x4.1x5.1 cm pituitary non-secreting macroadenoma. He had a ten year history of gradual loss of vision in the right eye, six month history of fatigue, and one month history of low libido. According to the MRI, the mass significantly compressed the optic chiasm and surrounded the internal carotid arteries. His laboratory analysis showed central hypogonadism (FSH 1.0 mIU/mL (1.5-12.4), LH 1.6 mU/mL (1.7-8.6), testosterone 13.0 ng/dL (249-836)), central hypothyroidism (TSH 4.54 mIU/L (0.5-5.7), Free T4 0.5 ng/dL (0.9-1.7)),central adrenal insufficiency (ACTH 15 pg/mL (10-60), random cortisol 1.8 ug/dL (2.3-11.9)), and growth hormone deficiency (growth hormone 0.01 ng/mL (0.01-0.97), somatomedin C 27 ng/mL (91-246)). He was started on levothyroxine 100 mcg daily and dexamethasone 2 mg bid, however the patient did not take the prescribed medications. Following the transphenoidal resection of the mass, he was started on hydrocortisone 50 mg IV bid. On post-operative day 3, one hour after infusion of the hydrocortisone, the patient developed symptoms of acute psychosis. He was disoriented, combative, chanting religious incantations, and swinging at the staff. On post-operative day 4, steroids were decreased to hydrocortisone 20 mg in the morning and 10 mg in the evening. He remained psychotic and had episodes of alternating waking and losing consciousness with stable vital signs. A repeat CT scan of the head did not show any change from the initial post-operative scan. He refused the steroids following post-operative day 5. Subsequently, his mental status cleared on post-operative day 7 and his repeat morning cortisol was 12.5 ug/dL and 11.6 ug/dL off steroids on post-operative days 8 and 9, respectively. He was discharged on levothyroxine and no steroids.[br][bold]Conclusion:[/bold] Patients with prolonged adrenal insufficiency, both primary and secondary, are at risk for acute psychosis following initiation of corticosteroids.[br][br]Nothing to Disclose: ESL, CB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 878 75 794 SAT-716 PO55-02 Saturday 733 2012


734 ENDO12L_SAT-717 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Nonfunctioning Pituitary Macroadenoma: 14-Year Follow-Up of a Neglected Case Vafa Tabatabaie, Rita J Louard Montefiore Medical Center, Bronx, NY Background[br]Long term monitoring of nonfunctioning benign pituitary macroadenomas is emphasized in literature, as they can cause hypopituitarism and visual field defects even if they are asymptomatic at presentation. We present the case of a macroadenoma that caused significant morbidity 14 years after initial detection.[br]Clinical Case[br]Patient is a 43-year-old woman who presented to emergency department complaining of worsening headache, nausea and vision loss. She also reported increasing fatigue and 80 lbs weight gain over the course of the past year. Menstrual history was unavailable due to bilateral oophorectomy secondary to torsion 5 years ago. MRI of the brain revealed a 3.5 x 2.5 x 2.5 cm sellar mass with displacement of optic chiasm and optic nerves as well as abutment of cavernous sinus and anterior cerebral arteries. Laboratory evaluation revealed panhypopituitarism (FT4 0.65 ng/dl; TSH 0.81 uU/ml; LH 2.9 mIU/ml; FSH 25.3 mIU/ml; IGF-1 81 ng/ml; cortisol 3.7 ug/dl) and a normal prolactin (29.5 ng/ml). Visual field exam revealed superior bitemporal loss. Patient subsequently reported that 14 years ago during work up for recurrent headaches she was told she has a pituitary tumor in another facility. Review of records obtained from the other hospital revealed a 12mm non-enhancing pituitary lesion and normal hormonal values. However, she failed to return for follow up imaging or hormonal evaluation as recommended until the current presentation. She underwent an uneventful trans-sphenoidal excision of the adenoma. Pathology confirmed a pituitary adenoma with positive staining for FSH and LH, with a minor component of prolactin and GH. One month after surgery, patient reported improved energy and moderate weight loss on hormonal replacement; Her visual fields and headaches have improved.[br]Clinical lessons[br]Despite the lack of evidence from randomized studies, most experts agree that in the absence of visual field defect, expectant management with regular follow up is an acceptable therapeutic approach for nonfunctioning pituitary macroadenomas. The present case underscores the importance of long-term regular follow up, with hormonal and imaging studies, if an expectant management is chosen. Examples of macroadenomas followed beyond 10 years is rare in literature. This case illustrates the potential for significant growth causing visual morbidity, as well as hypopituitarism, as long as 14 years after initial diagnosis.[br][br](1) M. Merc[egrave] Fern[aacute]ndez-Balsells, Mohammad Hassan Murad, Amelia Barwise, Juan F. Gallegos-Orozco, Anu Paul, Melanie A. Lane, Julianna F. Lampropulos, In[eacute]s Natividad, Lilisbeth Perestelo-P[eacute]rez, Paula G. Ponce de Le[oacute]n-Lovat[oacute]n, Patricia J. Erwin, Jantey.[br][br]Nothing to Disclose: VT, RJL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 528 75 795 SAT-717 PO55-02 Saturday 734 2012


735 ENDO12L_SAT-718 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Pineal Gland Tumor and Panhypopituitarism in an Adult Male: A Case Report and Review of Endocrine Manifestations Queenie Guinto Ngalob, Wilfredo Liangco, Gabriel Jasul University of the Philippines-Philippine General Hospital, Manila, Philippines; University of the Philippines-Philippine General Hospital, Manila, Philippines Background: Tumors of the pineal gland are rare and occur in children or young males in the first two decades of life. The most common histologic type is germ cell tumor. It usually comes to medical attention due to mass effect. Endocrine manifestations are also prominent in the clinical presentation and may precede neurologic symptoms.[br]Case: We report a 30 year old male with a pineal gland tumor who presented with six months history of headache, generalized weakness, anorexia, weight loss, polyuria and increased thirst. History revealed that he had thinned facial hair, loss of axillary hair, poor libido and lack of morning erections even earlier. Few days onset of doubling of vision, vomiting and disorientation prompted medical consult. Pertinent in the physical examination was limitation of vertical movements of both eyes. Imaging showed a pineal gland tumor with obstructive hydrocephalus and an unremarkable sellar region. Serum AFP and [beta]HCG were both elevated. A presumptive diagnosis of nongerminomatous germ cell tumor was made. Hormonal panel demonstrated central adrenal insufficiency, central hypothyroidism, hypogonadotropic hypogonadism and mild hyperprolactinemia. Water deprivation test revealed diabetes insipidus. Treatment was multimodal. Insertion of a ventriculoperitoneal shunt was done to relieve the hydrocephalus. Hormone replacement with prednisone, levothyroxine and desmopressin were given. Cranial irradiation was done which resulted in more than 50% regression of the tumor size. There was also resolution of limitation of eye movements. Three months later, he developed bilateral leg numbness and weakness which rapidly progressed to paralysis within a few days. Spinal metastasis was demonstrated on magnetic resonance imaging. Spinal irradiation was done which yielded some recovery in sensation but he remained paraplegic. He also underwent chemotherapy using cisplatin, bleomycin and etoposide but was put on hold due to infection after the first cycle.[br]Conclusion: The characteristic clinical triad of diabetes insipidus, anterior pituitary failure and visual disturbances is a harbinger of a tumor in the pineal region. Pineal germ cell tumors may present with pituitary dysfunction even in the absence of pituitary involvement on imaging and is considered disseminated disease. Thorough endocrine evaluation and hormone replacement is essential in its management.[br][br]Nothing to Disclose: QGN, WL, GJ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1083 75 796 SAT-718 PO55-02 Saturday 735 2012


736 ENDO12L_SAT-719 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) A 28-Year-Old Master[apos]s Student with Untreated Congenital Panhypopituitarism Mariana Costa Silva, Marcelo Fernando Ronsoni, Cristina Schreiber Oliveira, Maria Heloisa Busi Silva Canalli, Marisa Helena Cesar Coral, Alexandre Hohl University Hospital - Federal University of Santa Catarina, Florian[oacute]polis, Brazil [bold]Background[/bold]: The incidence of Congenital Hypopituitarism (CH) is estimated to be between 1:3000-4000 births. Clinical presentation is variable, depending on the type and severity of deficiencies and on the age at diagnosis. If untreated, main symptoms include short stature, cognitive alterations, delayed puberty. An adequate treatment for hormone deficiencies and a strict follow-up are necessary.[br][bold]Case presentation:[/bold] Male, 28 y.o., black, master[acute]s student in engineering, coming from Mozambique, 135cm tall, weight 41kg, no signs of virilization, Turner stage P1G1 (penis 3cm long, testis 3cm[sup]3[/sup]). The man had always been short for his age but denied learning difficulties, head injury, radiotherapy, any other health problem and any family member with similar characteristics. He reported libido and erections, but never had sex and that was a complaint. He was taking Sinvastatin 20mg daily since 3 months earlier. Bone age was compatible with 15 years. Laboratory tests: TSH 2.5[mu]IU/mL (0.4-4.0), FT4 0.64ng/dL (0.89-1.76), LH 0.16mIU/mL (0.8-7.6), FSH 0.65mIU/mL (0.7-11.1), prolactin 25.2ng/mL (2.5-17), GH[lt]0.005ng/mL, IGF1[lt]25ng/dL (117-329), total testosterone[lt]20ng/dL, SHBG 34.4nmol/L (13-71), 8:00AM cortisol 11.3[micro]g/dL (5-25), fasting blood glucose 66mg/dL, hemoglobin 13.5g/dL, total cholesterol 298mg/dL, HDL 41mg/dL, LDL 197mg/dL, triglycerides 167mg/dL. In the insulin tolerance test, 30min. after administration of 2IU of insulin, glucose was 28mg/dL, cortisol was 10.6 [micro]g/dL and GH was 0.07ng/mL. MR imaging revealed discrete arachnoidocele intrasellar, small anterior pituitary, neurohypophysis was unidentified, marked narrowing of the pituitary stalk. After the endocrinology evaluation he had started hormone replacement therapy: Prednisone 2.5mg daily, levothyroxine 25mcg daily, somatotropin 6UI a day and topic dihidrotestosterone on the penis area once a day. After 9 months of treatment, the patient shows an increase of 5cm height and a penile growth of 4cm, his weight was 43.5kg (BMI 22.2), total cholesterol 214, HDL 59, LDL 132, triglycerides 71. [bold]Conclusion: [/bold]This case shows the natural history of an adult with an untreated CH and the short-term outcome of hormonal replacement therapy. It[acute]s interesting that he didn[acute]t have cognitive deficits despite severe hypothyroidism, leading us to think that it was a new-onset hypothyroidism. His 9 months response in the growth curve, penis size and in lipids profile is encouraging in terms of psychosocial and health benefits.[br][br]Nothing to Disclose: MCS, MFR, CSO, MHBSC, MHCC, AH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 714 75 797 SAT-719 PO55-02 Saturday 736 2012


737 ENDO12L_SAT-720 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Untreated Congenital Combined Pituitary Hormone Deficiency in a 69-Year-Old Man Payal Satish Patel, Steven G Waguespack Baylor College of Medicine, Houston, TX; University of Texas MD Anderson Cancer Center, Houston, TX Introduction: Congenital combined pituitary hormone deficiency (CPHD) can stem from perinatal trauma, developmental CNS defects, or mutations in pituitary transcription factors which can be associated with anatomical defects such as a thin or interrupted pituitary stalk, ectopic or absent posterior pituitary, and/or hypoplasia of the anterior pituitary. A diagnosis of congenital CPHD in adulthood is exceedingly rare.[br]Clinical Case: A 69-year-old man with stage III melanoma was referred for evaluation after MRI brain revealed a prominent pituitary gland and lack of visualization of the pituitary stalk. His parents initially became concerned when, at the age of 7, he became shorter than his younger brother. He failed to enter puberty and never married or fathered children. He had seen different specialists in Mexico as a child and transiently took thyroid hormone replacement. Once moving to the U.S., he did not seek additional treatment. He denied any history of head trauma or any unusual circumstances regarding his birth. He had sustained a low impact hip fracture at the age of 40. Review of systems was pertinent for cold intolerance, fatigue, and occasional constipation. Exam was pertinent for a man of short stature (154cm) with hypogonadal, triangular facies. He had a high-pitched voice and lacked body hair. There was no gynecomastia and his trunk and abdomen were youthful in appearance. Penis was not fully developed and testes were retractile and estimated to be 4cc in volume.[br]Laboratory results were as follows: TSH 1.57 uIU/mL (0.27-4.20), Free T4 0.54 ng/dL (0.93-1.70), LH [lt]0.7 mIU/mL (1.7-8.6), FSH 0.1 mIU/mL (1.5-12.4), Free testosterone [lt]0.2 ng/dL (40-168), total testosterone [lt]7 ng/dL (240-950), prolactin 453 ng/mL (4-15.2), 6am cortisol 18.8 mcg/dL, 6am ACTH 7 pg/mL ([lt]46), IGF-1 32 ng/mL (67-195), sodium 140 mEQ/L. DXA scan revealed osteoporosis with a T-score of -4.8 at the left hip and -3.2 at the lumbar spine.[br]Conclusion: Congenital CPHD is usually diagnosed in infancy or childhood due to typical signs and symptoms such as neonatal hypoglycemia, prolonged jaundice, and/or micropenis as well as growth retardation during childhood. To date, there are less than 10 case reports of untreated congenital CPHD in adults, most of whom were [lt]50 years of age at time of diagnosis. This case demonstrates the natural history and potential complications from untreated CPHD associated with an interrupted pituitary stalk.[br][br]Nothing to Disclose: PSP, SGW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1130 75 798 SAT-720 PO55-02 Saturday 737 2012


738 ENDO12L_SAT-721 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Panhypopituitarism Due to Septo-Optic-Dysplasia Imad Brema, Eoin Fitzgerald, Donal O[apos]Shea St Vincent[apos]s University Hospital, University College Dublin, Dublin, Ireland Background: Septo-optic dysplasia (SOD) is a rare congenital condition characterised by the triad of optic nerve hypoplasia (ONH), midline brain abnormalities and panhypopituitarism. Phenotypically, SOD is a quite variable condition leading to delayed diagnosis and adverse outcomes.[br]We report a rare case of SOD in a 16 years old Irish Caucasian female who was referred by her family physician for evaluation of short stature. She was born at 34 weeks gestation following spontaneous pre-term labour. Shortly after birth, the patient was diagnosed with bilateral optic nerve hypoplasia and had two corrective surgeries for nystagmus and congenital squint in California, USA, before her 1[sup]st[/sup] birthday. The patient had primary amenorrhea and visual impairment and was always smaller than her peers and required additional assistance in school. Initial work up by her family physician confirmed central hypothyroidism with free T4 7.9 pmol/L (N: 12-22) and TSH 3.9 mIU/L (N: 0.25-4.0). The rest of her anterior pituitary hormones confirmed panhypopituitarism as follows: 9 am cortisol 65.4 nmol/L (N: 200-400); IGF-: 34 ug/L (N: 261-752); estradiol: 99 pmol/L (N: 130-500); LH: 4.6 U/L (N: 2-12); FSH: 8 U/L(N: 2-12). Prolactin was normal at 496 mIU/L (N: 105-548). Physical examination findings were as follows: height: 145 cm ([lt] 3[sup]rd[/sup] centile), weight: 45.0 kg (Just above the 3[sup]rd[/sup] centile), absent secondary sexual characteristics, and bilateral nystagmus. MRI Brain and pituitary showed agenesis of the septum pellucidum, thinning of the corpus callosum and a relatively small pituitary gland, confirming the diagnosis of SOD. X ray of the left wrist showed a slightly delayed bone age with complete closure of all phalangeal epiphyses and almost complete fusion of ulnar and radial epiphyses.[br]The patient was started on hydrocortisone, thyroid hormone, and GH replacements, however, her height only increased by 0.5 cm over 6 months despite achieving appropriate IGF-1 levels.[br]Conclusion: This case illustrates the challenge of diagnosing SOD at an earlier age, partly due to a wide phenotypic variation as well as late development of pituitary hormone deficiencies. However, the presence of ONH at birth should always prompt clinicians to suspect the diagnosis and do further investigations, such as brain imaging and regular pituitary hormone measurements. Late diagnosis of SOD exposes patients to complications including those of undiagnosed hypopituitarism and short stature.[br][br]Nothing to Disclose: IB, EF, DO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 709 75 799 SAT-721 PO55-02 Saturday 738 2012


739 ENDO12L_SAT-722 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Pituitary Abscess Presenting as Hypopituitarism in an Adolescent: Case Report and Review of the Literature Nancy Gagne, Julie Doyon, Cybele Bergeron, Chantal Lemire, Yves Patenaude Universit[eacute] de Sherbrooke, Sherbrooke, Canada; Universit[eacute] de Sherbrooke, Sherbrooke, Canada; Universit[eacute] de Sherbrooke, Sherbrooke, Canada Pituitary abscesses are rare causes of pituitary masses in general but especially rare in children with only 18 cases described to our knowledge in the literature to date. Few patients present with symptoms of infection, with most patients presenting with endocrine dysfunction and/or visual disturbance and headache. Treatment is transphenoidal removal of the abscess with or without antibiotics. Hypothalamic and pituitary evaluation pre and post surgery is warranted.[br]We describe a case of a 17 year old adolescent girl who presented with symptoms of polydipsia, polyuria, oligomenorrhea and weight gain suggesting a hypothalamic or pituitary lesion. Preoperative evaluation confirmed the presence of diabetes insipidus, central hypothyroidism, partial hypogonadotropic hypogonadism as well as a possible growth hormone deficiency with no evidence of hypothalamic-pituitary-adrenal axis disturbance. The preoperative diagnosis based on history, laboratory investigation and magnetic resonance imaging was that of a craniopharyngioma or a germinoma. After transphenoidal resection, pathology revealed that the resected mass was a well-organized pituitary abscess with no specific infectious agent found on staining. Cultures had not been taken at the time of surgery because of the appearance of the mass. After multiple investigations, the patient was not found to have any predisposing conditions for a pituitary abscess and she was not treated with antimicrobials. The only noteworthy finding on history was that she had been treated for a dental abscess one year prior to presentation but at no time had she presented with symptoms of an infectious intracranial process. Evolution post-surgery revealed the central hypothyroidism to be transient, but the diabetes insipidus and the hypogonadotrophic hypogonadism persist 2 years post surgery. The hypothalamic-pituitary adrenal axis remained normal throughout follow-up. IGF-1 levels, which were low pre-operatively increased to normal levels post-operatively suggesting somatotrope recovery of function.[br]We therefore conclude that a pituitary abscess should be part of the differential diagnosis of pituitary masses in children even when no infectious symptoms are present and that tissue should be set aside for cultures during surgery when possible. Although most patients do well with removal of the abscess alone many patients remain with endocrine deficits which require follow-up and treatment.[br][br]Nothing to Disclose: NG, JD, CB, CL, YP 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1851 75 800 SAT-722 PO55-02 Saturday 739 2012


740 ENDO12L_SAT-723 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Recurrent Pericardial Effusion and Shock Due to Hypothyroidism and Adrenal Crisis: A Case of Unrecognized Sheehan Syndrome Jongoh Kim, Ramona Dadu, Ramaswami Nalini Baylor College of Medicine, Houston, TX [bold]Background:[/bold] Sheehan[apos]s syndrome (infarction of the pituitary gland after post-partum hemorrhage) is usually recognized from inability to lactate and failure to resume menses after delivery. When the hypopituitarism is mild there may be a delay in recognition for several years. We present a case where it was recognized after recurrent adrenal crisis.[br][bold]Case:[/bold] A 41 year old Hispanic woman with long history of hypothyroidism, non-compliant on levothyroxine replacement therapy, presented with pleuritic chest pain and shortness of breath. She had history of recurrent pericardial and pleural effusion in the past few years with hypotension and shock, without tamponade. Six months prior to presentation, she underwent pericardial window for increasing pericardial effusion. Pathology and microbiology evaluation showed no evidence of bacterial or fungal infection, tuberculosis, HIV, malignancy, or rheumatologic disorders. At the time of presentation to our institution, her BP was 78/47 mmHg requiring vasopressor support. Review of the history revealed that she had been amenorrheic for 20 years since the delivery of her daughter. Delivery was complicated by heavy bleeding requiring transfusion and she was unable to lactate. She had also lost her body, axillary, and pubic hair gradually. Laboratory evaluation revealed adrenal insufficiency based on inadequate response of cortisol to 250 mcg Cosyntropin (synthetic ACTH): basal cortisol level was 3.7 ug/dL and peak response was 6.2 ug/dL (range: [gt]18 ug/dL). Serum free T4 and TSH levels were low: 0.19 ng/dL(range 0.89-1.76 ng/dL) and 0.42 uIU/mL (range:0.35 -5.50 uIU/mL) respectively. Serum gonadotropins, estradiol, prolactin, and IGF-1 levels were all low. MRI brain did not show any abnormalities in the pituitary fossa. She received treatment with high dose steroids with improvement in blood pressure. She was diagnosed with panhypopituitarism secondary to Sheehan[apos]s syndrome. She was discharged in a stable condition with replacement doses of hydrocortisone and levothyroxine. She has remained stable without further hospitalization.[br][bold]Clinical Lesson:[/bold] Our patient had poorly controlled hypothyroidism which was the cause of recurrent pericardial effusion and presented with hypotension due to adrenal crisis. Recognition of panhypopituitarism due to Sheehan[apos]s syndrome in women with hormonal deficiencies by a detailed history is important to avoid significant morbidity and mortality.[br][br]Nothing to Disclose: JK, RD, RN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1669 75 801 SAT-723 PO55-02 Saturday 740 2012


741 ENDO12L_SAT-724 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Pituitary Apoplexy Secondary to Dengue Hemorrhagic Fever: An Under-Recognized Complication? Seng Kiong Tan, Cherng Jye Seow, Rinkoo Dalan Tan Tock Seng Hospital, Singapore, Singapore [bold][underline]Introduction[/underline][/bold]: Pituitary apoplexy is a rare life threatening emergency. We report a case in a gentleman, who presented with pituitary apoplexy precipitated by thrombocytopenia from dengue fever.[br][bold][underline]Case Presentation[/underline][/bold]: A 53 years man presented with fever, headache and lethargy for 5 days. He had left complete ptosis, right sixth nerve palsy and right temporal hemianopia. MRI pituitary revealed a 3.7 x 2.4 x 3.1 cm hemorrhagic pituitary macroadenoma with bilateral cavernous sinus extension and optic chiasm compression. Significant laboratory findings: Platelet 16000/uL, dengue IgM and IgG positive, Prolactin [gt]10000 mIU/L (RI: 80.0-390.0 mIU/L), Cortisol (0min) 127 nmol/L, fT4 12.24pmol/L (RI:10.00-20.00 pmol/L), TSH 0.246 (RI:0.400-4.00 mIU/L), Total testosterone 0.651 nmol/L (RI: 9.900-27.800 nmol/L). After initial intravenous dexamethasone and platelet transfusion, he underwent a trans-sphenoidal evacuation. Although there was a transient improvement initially, his right eye vision deteriorated again. A repeat MRI showed a recurrent hemorrhagic pituitary mass. A re-exploration surgery was performed to evacuate the hematoma. Histology confirmed a necrotic prolactin secreting pituitary adenoma. He was continued on cabergoline and hydrocortisone. A repeat MRI 3 months later showed residual tumor in the left sella with bilateral cavernous sinus extensions. Although prolactin concentrations normalized, panhypopituitarism and right eye visual defects persisted.[br][bold][underline]Discussion[/underline][/bold]: Pituitary apoplexy has been reported in acute thrombocytopenic states that can occur in ITP (1), after chemotherapy in acute myeloid leukemia (2) or administration of thrombolytic agents, (3)or anticoagulant drugs (4). However a literature review revealed only one case of dengue fever complicated by hemorrhage in a pre-existing pituitary adenoma with platelet count of 45000/ul (5). Although, our case and the case reported by Kumar et al, did not have any other systemic hemorrhage, they developed pituitary apoplexy with significant sequelae. Hence, thrombocytopenia from any cause is liable to precipitate pituitary apoplexy.[br][bold][underline]Conclusion[/underline][/bold]: Although pituitary apoplexy from thrombocytopenia due to dengue fever is uncommon, a high index of suspicion is required when patients with thrombocytopenia develop headache and visual field changes.[br][br](1) Maiza J C, Bennet A, Thorn-Kany M., et al. Pituitary apoplexy and idiopathic thrombocytopenic purpura : a new case and review of the literature. Pituitary 2004; 7(3): 189-92. (2) Silberstein L, Johnston C, Bhagat A, Tibi L et al. Pituitary apoplexy during induction chemotherapy for acute myeloid leukemia. Br J Haematol 2008; 143(2):151. (3) Hyer S, Soo S, Taylor W, Nassey S. Spontaneous hemorrhage into pituitary tumor after streptokinase therapy. Postgrad Med J 1993;69:244. (4) Oo M, Krishna A, Bonavita G, Rutecki G. Heparin therapy for myocardial infarction; an unusual trigger for pituitary apoplexy. Am J med Sci 1997;314:351-3. (5) Kumar V, Kataria R, Mehta VS. Dengue hemorrhagic fever: a rare case of pituitary tumor hemorrhage and reversible vision loss. Indian J Opthalmol 2011;59:311-2.[br][br]Nothing to Disclose: SKT, CJS, RD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 139 75 802 SAT-724 PO55-02 Saturday 741 2012


742 ENDO12L_SAT-725 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Apoplexy of a Microprolactinoma during Pregnancy Nathalie Couture, Nahla Aris-Jilwan, Omar Serri Centre Hospitalier de l[apos]Universit[eacute] de Montr[eacute]al, Montr[eacute]al, Canada [bold]Case report:[/bold] A 37-year-old woman with a history of a microprolactinoma presented at the 16th week of her first pregnancy, with a sudden onset of severe headache, nausea, vomiting and blurred vision. She had been receiving first bromocriptine for 8 months and then switched to cabergoline for over 6 months until she became pregnant. Magnetic resonance imaging (MRI) revealed a sellar heterogeneous mass with suprasellar extension (11x11x18 mm) and contact with the optic chiasm compatible with adenoma apoplexy. A previous MRI performed 4 months prior to conception showed a 7x7 mm microadenoma. Upon referral to our center, the patient[apos]s pituitary functions and visual fields were assessed and found to be normal. Cabergoline treatment was reinitiated at 0.5 mg per week with subsequent resolution of the headache and nausea. An MRI performed five weeks later showed a marked regression of the sellar mass to 3x7x15 mm. At subsequent follow-up visits 20th and 24th weeks of pregnancy, an obstetrical ultrasound showed normal fetus development and the patient remained asymptomatic. Cabergoline was discontinued at 34th week of pregnancy, four weeks before the expected delivery date. At the 38th week of pregnancy the patient was admitted in the obstetrics ward with onset of labor. Because of mild preeclampsia and failed induction, she delivered after a cesarean section an alive and healthy female baby with a weight of 2.2 Kg. Normal lactation was gradually established after birth. A repeat MRI was performed 1 week after delivery and showed no progression of the mass or new apoplexy. The patient is doing well 2 months postpartum. She is breastfeeding intermittently and her prolactin level is stable at 24 [mu]g/L without treatment.[br][bold]Discussion:[/bold] During pregnancy, estrogen secretion promotes the growth of the normal pituitary gland. Microprolactinomas rarely show a symptomatic increase in size during pregnancy. A review by Molitch showed that clinically significant growth may occur in 2.7% of cases. Apoplexy of microadenomas has not been previously described.[br][bold]Conclusion:[/bold] This case illustrates an unusual behaviour of a microprolactinoma with a rapid progression and apoplexy during pregnancy. Reinitiation of cabergoline treatment was needed to prevent a deleterious increase of adenoma or normal pituitary size. The successful pregnancy allowed for the delivery of a heathy child.[br][br]Molitch ME. Prolactinoma in pregnancy. Best Pract Res Clin Endocrinol Metab. 2011 Dec;25(6):885-96.[br][br]Nothing to Disclose: NC, NA-J, OS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 432 75 803 SAT-725 PO55-02 Saturday 742 2012


743 ENDO12L_SAT-726 POSTER SESSION: Vasopressin Deficiency [amp] Excess [amp] Hypopituitarism Case Reports (1:30 PM-3:30 PM) Pituitary Apoplexy with Cabergoline Therapy: Case Report and Literature Review Edwin Chng, Rinkoo Dalan Tan Tock Seng Hospital, Singapore, Singapore [bold]Introduction: [/bold]Pituitary apoplexy is a rare medical emergency which results from hemorrhage or infarction in the pituitary gland. Although pituitary apoplexy is known to occur after treatment with bromocriptine, very few cases have been reported after initiation of cabergoline. We report a case of a gentleman with macroprolactinoma who developed pituitary apoplexy 6 weeks after initiation of cabergoline.[br][bold]Case presentation[/bold]: A 20 years old gentleman presented to the neurologist with a history of chronic headache for 5 years. A MRI of the pituitary revealed a large sellar cystic macroadenoma with suprasellar extension. Serum prolactin was raised at 60394 mIU/L (RI: 77-274) while the rest of his hormonal profile was normal suggestive of a macroprolactinoma. He was started on cabergoline 0.25 mg twice a week which was uptitrated to 0.5 mg twice a week after 4 weeks. Six weeks after initiation with cabergoline he presented to the emergency department with severe headache, associated with nausea and vomiting. An urgent MRI confirmed pituitary apoplexy. Since he did not have any neurological deficits he was treated conservatively with complete recovery. Cabergoline was continued as he still had high prolactin levels but he did not develop any other deficiencies.[br][bold]Discussion: [/bold]Biller et al reported the first case in 1995 (1). Six more cases have been reported after that (2-5). A review of all these 7 cases shows that apoplexy has occurred as early as 1 week and as late as 12 months after initiation with cabergoline and has occurred even with the minimum dose of 0.5 mg/week. Although the exact mechanism by which these drugs precipitate apoplexy is not known, it is known that dopamine agonists cause apoptosis of lactotrophe cells with a decrease in metabolic demands and also inhibits angiogenesis of the tumor. An imbalance between the two factors with marked inhibition of angiogenesis can lead to infarction of tumor cells with resultant hemorrhagic necrosis.[br][bold]Conclusion:[/bold] Pituitary apoplexy is a rare complication that can develop in a pituitary macroprolactinoma as early as 1 week after initiation of cabergoline at even the minimum doses.[br][br](1) Biller BMK, Molitch ME, Vance ML, et al. Treatment of prolactin-secreting macroadenomas with the once-weekly dopamine agonist cabergoline. J Clin Endocrinol Metab 1996; 81: 2338-2343. (2) Vella A, Young Jr WF. Pituitary apoplexy. The Endocrinologist 2001;11:282-288. (3) Knoepfelmacher M, Gomes MC, Melo ME, Mendonca BB. Pituitary apoplexy during therapy with cabergoline in an adolescent male with prolactin-secreting macroadenoma. Pituitary. 2004; 7:83-87. (4) Lima GAB, Machado EOM, Silva CMS, Filho PN, Gadelha MR. Pituitary apoplexy during treatment of cystic macroprolactinomas with cabergoline. Pituitary. 2008;11:287-292. (5) Stojanovic ND, Gunganah K, Vidyarti M, Chawda S, Khatami Z, Pollock J. Management of a woman with giant invasive prolactinoma and pituitary apoplexy- a multidisciplinary effort. Endo 2011 Poster.[br][br]Nothing to Disclose: EC, RD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 97 75 804 SAT-726 PO55-02 Saturday 743 2012


1233 ENDO12L_SUN-413 POSTER SESSION: Thyrotoxicosis, TSH Administration [amp] Thyroid Cell Biology (1:30 PM-3:30 PM) Effects of 0.1 mg Recombinant TSH Radioiodine Therapy in Thyroidectomized Patients with Recurrent Multinodular Goiter Nikolaos G Angelopoulos, John Iakovou, Anastasios Boniakos Nuclear Medicine, Thessaloniki, Greece [bold]Background:[/bold] Several treatment options exist after thyroid malignancy has been ruled out in patients with multinodular goiter (MG). Surgery efficiently reduces the goiter size but carries a risk of both surgical and anesthetic complications while in recent years, levothyroxine suppressive therapy has been abounded. I131 therapy is the only nonsurgical alternative; however, the effectiveness diminishes with increasing goiter size and depends on iodine sufficiency in some areas. Recombinant human (rh) TSH approximately doubles the thyroid 131I uptake in patients with nodular goiter. Several studies have proved efficacy of rhTSH stimulated 131I therapy on goiter reduction. [bold]Objective:[/bold] The objective of the study was to assess the efficacy and safety of 0.1 mg rhTSH as an adjuvant to a fix dose of 131I therapy (11 mci) in patients with recurrence of large multinodular goiter several years after the initial thyrodectomy. [bold]Patients and Intervention:[/bold] 14 (13 females), age 59.1 (35-78) received 11mciu of I131, 24h after the administration of 0.1 mg rhTSH. [bold]Main Outcome Measures:[/bold] The primary end point was a change in thyroid volume (by ultrasound measurements) as well as in the diameter of the predominant nodule during a follow up period of 8 months. Secondary end points were the alterations in thyroid function and potential adverse effects. [bold]Results:[/bold] Significant decrease in the volume of thyroid remnant was observed at the end of the follow up period (log10 values), mean (95%CI) 1.17(1.03-1.3) after 8 months, compared to 1.45 (1.32-1.59) before. There was a positive correlation between the increments of TSH increase (TSH 24h after the stimulation -TSH before stimulation) with the percentile of thyroid volume decrease. (r=0.77, p[lt]0.01) [bold]Conclusion:[/bold] Further investigation is needed since this approach could be attractive in terms of minimizing the potential risks of reoperation in these patients.[br][br]Nothing to Disclose: NGA, JI, AB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1212 191 1440 SUN-413 PO39-02 Sunday 1237 2012


1234 ENDO12L_SUN-414 POSTER SESSION: Thyrotoxicosis, TSH Administration [amp] Thyroid Cell Biology (1:30 PM-3:30 PM) Inhibited [sup]131[/sup]I Uptake but Normal Release of Thyroid Hormones by Thyroid Gland in Response to TSH Administration in Subclinical Hypothyroidism Udaya Manohar Kabadi Unversity of Iowa, Iowa City, IA; VA Medical Center, Des Moines, IA [bold]Background:[/bold] Subclinical hypothyroidism (SH) is a syndrome characterized by normal thyroid hormone with supernormal TSH levels. In most subjects, a known etiologic factor is identified and therapy with levothyroxine normalizes serum TSH while maintaining normal thyroid hormone levels. However, the exact pathophysiology of these thyroid hormone alterations is not defined.[br][bold]Objective:A[/bold] major step in synthesis i.e. iodine uptake and the release of thyroid hormones in response to administration of TSH were assessed in subjects with SH.[br][bold]Subjects and Methods:[/bold] 10 men and 5 women with ages 42-76 years with SH secondary to Hashimoto[apos]s thyroiditis and 10 euthyroid men (N)with ages 39-70 years participated in the study. 24 hr [sup]131 [/sup]I thyroid uptake and serum TSH, T[sub]4[/sub] and T[sub]3[/sub] were determined before and after SC administration of recombinant human TSH, 0.9 mg for two consecutive days. Comparisons were conducted between subjects with SH and N.[br][bold]Results:[/bold] In subjects with SH, 24 hour [sup]131[/sup]I thyroid uptakes were within normal range [10-30%] as defined by local nuclear medicine laboratory. However, the mean value (13[plusmn]2)was significantly lower [p[lt]0.05] as compared to N(25[plusmn]4). 24 hr uptake values rose following TSH administration in both groups. However, the rise was significant [p[lt]0.01] only in N. Furthermore, both the mean absolute uptake value and the mean % rise following TSH administration from preTSH uptake value were significantly lower in subjects with SH (16[plusmn]3 and 22[plusmn]3) in comparison to N(51[plusmn]9 and 80[plusmn]10).Serum T[sub]3[/sub] and T[sub]4[/sub] concentrations in subjects with SH were within normal range(80-180 ng/dl; 4.5-11.0 ug/dl) as established by the clinical laboratory. Moreover, the mean levels (136[plusmn]11 and 8.3[plusmn]0.7) were also not significantly different in comparison to N(141[plusmn]12 and 8.4[plusmn]0.6). Basal Serum TSH concentrations were supernormal and significantly higher[p[lt]0.05] in subjects with SH(8[plusmn]2)in comparison to N(2[plusmn]1) and rose markedly in both groups following TSH administration with no significant difference among the elevations between groups. Serum T[sub]4[/sub] and T[sub]3[/sub] rose significantly from PreTSH levels in both groups (p[lt]0.05). However, the rises were not significantly different between groups (T[sub]4 [/sub]ug/dl,10.2[plusmn]1.1in SH Vs10.4[plusmn]1.2in N; T[sub]3 [/sub]n g/dl, 166[plusmn]17in SH vs.170[plusmn]16 in N)[br][bold]Conclusion: [/bold]In subjects with SH secondary to Hashimoto[apos]s thyroiditis, 24 hour [sup]131 [/sup]I Thyroid uptake is inhibited prior to as well as following SC TSH administration in comparison to N with maintainance of normal hormone release.[br][br]Sources of Research Support: Department of Veterans Affairs.[br][br]Nothing to Disclose: UMK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 254 191 1441 SUN-414 PO39-02 Sunday 1238 2012


1235 ENDO12L_SUN-415 POSTER SESSION: Thyrotoxicosis, TSH Administration [amp] Thyroid Cell Biology (1:30 PM-3:30 PM) Efficacy of Administering 0.45 mg of Recombinant Human TSH for Two Consecutive Days on Stimulating TSH and Thyroglobulin in Patients Treated for Differentiated Thyroid Carcinoma Lia Monte, Elaine Oliveira Dias, Eduardo Kiyoshi Tomimori, Meyer Knobel, Rosalinda Asato Camargo S[atilde]o Paulo University School of Medicine, S[atilde]o Paulo, Brazil Background: Recombinant human thyrotropin (rhTSH) stimulation is frequently used to asses the disease status of patients treated for differentiated thyroid carcinoma (DTC) with total thyroidectomy and ablative doses of 131I. It is also used when basal (unstimulated) thyroglobulin (b-TG) is below the assay sensitivity limit. Objective: The aim of this study was to evaluate the efficacy of administering 0.45 mg of recombinant human TSH (rhTSH) for two consecutive days in stimulating TSH and evaluate the 72-hour rhTSH-stimulated Tg (rhTSH-Tg) in patients submitted to total thyrodectomy for DTC. Methods: fifty-four adult patients (46 female and 8 male) were studied after rhTSH administration at a dose of 0.45 mg i.m. for 2 consecutive days. Serum basal thyroglobulin (Tg), TSH and Tg antibody (TgAb) measurement were performed before rhTSH administration. Stimulated TSH was measured 24 hs after the second dose of rhTSH and rhTSH-Tg 72 hs after the second dose. Patients were divided into 2 groups depending on serum Tg concentrations on suppressed TSH (THST): 48 patients had basal Tg concentrations of [lt] 1ng/mL (group A) and 6 patients had basal Tg values of [gt] 1ng/ml (group B). Results: TSH values increased [gt]30mU/mL in all patients after rhTSH, 85% had TSH [gt]60mU/mL and mean basal TSH values increased from 0.33 mU/mL to 84.97mU/mL (range 35.8 [ndash] 546.1mU/mL). In group A, Tg values remained at [lt]1ng/mL in 33 patients and increased from [lt]1ng/mL to 3.55ng/mL (median) in 15 patients after rhTSH administration. In group B, Tg values increased in all 6 patients from 8.46ng/mL(median) to 26.03ng/mL (median). No major adverse effects were reported after rhTSH administration. Conclusion: We concluded that the two doses of 0.45 mg of rhTSH are able to stimulate TSH and Tg.[br][br]Nothing to Disclose: LM, EOD, EKT, MK, RAC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2353 191 1442 SUN-415 PO39-02 Sunday 1239 2012


1236 ENDO12L_SUN-416 POSTER SESSION: Thyrotoxicosis, TSH Administration [amp] Thyroid Cell Biology (1:30 PM-3:30 PM) Mitochondrial OxPhos Defect in Thyroid Epithelial Cells Results in Mislocalization of NIS and Functional Failure Kyong Hye Joung, Jung Uee Lee, Min Hee Lee, Seong Eun Lee, Min Jeong Choi, Min Jeong Ryu, Soung Jung Kim, Young Suk Jo, Minho Shong Chungnam National University Hospital, Daejeon, Korea; Daejeon St Mary[apos]s Hospital, The Catholic University of Korea, Daejeon, Korea [bold]Background;[/bold] Mitochondria could regulate endocrine cell function with cell-type specific manner, such as insulin secretion in beta cells and aldosterone biosynthesis in glomerulosa cells. However, thyroid-follicular cell specific mitochondrial function has not been investigated. In this study, we have developed the new animal model of thyroid-specific mitochondrial dysfunction using standard gene targeting technology to understand the role of mitochondria in thyroid follicular cells.[br][bold]Results;[/bold] We analyzed the phenotypes of the mice with mitochondrial dysfunction generated by the defect of mtDNA-coded OxPhos complex. Homozygote thyroid-specific mitochondrial dysfunction (ThyCKO) mice retarded growth and died prematurely in PN21 to 35 days with severe thyroid dysfunction. Histological analyses of homozygote ThyCKO mice showed distortion of thyroid follicles and flattened follicular cells. Heterozygote ThyCKO mice showed apparently normal thyroid endocrine function. However, histological examination in heterozygote ThyCKO mice exhibited swollen follicular cells, which were proven to be related to endoplasmic reticulum (ER) swelling by electro-microscopic study. Interestingly, immunohistochemical study presented the failure of NIS to membrane targeting. Furthermore, short-term or prolonged TSH injections to heterozygote ThyCKO mice could exacerbate failure of NIS to membrane targeting with profound ER swelling, resulting in decreased thyroid hormone synthesis derived by TSH.[br][bold]Conclusion;[/bold] Our observational data suggested that sub-phenotypical mitochondrial OxPhos defect could cause structural and functional changes of thyroid follicular cells, which was able to determine follicular cell responsiveness to TSH.[br][br]Sources of Research Support: This work was supported in part by the second phase of the Brain Korea 21 program of the Ministry of Education and by the Korea Healthcare technology R[amp]D Project, MHW(A100588). YSJ [amp] MS were supported by NRF/MEST (2010-0020527).[br][br]Nothing to Disclose: KHJ, JUL, MHL, SEL, MJC, MJR, SJK, YSJ, MS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 68 191 1443 SUN-416 PO39-02 Sunday 1240 2012


1237 ENDO12L_SUN-417 POSTER SESSION: Thyrotoxicosis, TSH Administration [amp] Thyroid Cell Biology (1:30 PM-3:30 PM) Accumulation of Intracellular Lipids and Induction of Lipogenic Responses in Thyroid Gland of Obese Animals and Humans Min Hee Lee, Jung Uee Lee, Min Jeong Choi, Min Jeong Ryu, Soung Joong Kim, Yong Kyung Kim, Koon Soon Kim, Minho Shong, Young Suk Jo Chungnam National University Hospital, Daejeon, Korea; Daejeon St Mary[apos]s Hospital, The Catholic University of Korea, Daejeon, Korea [bold]Background; [/bold]Ectopic fat deposition in non-adipose tissue is a common health problems e.g., fatty liver in obesity syndrome. Although it has long been studied that ectopic fat accumulation and lipogenic activation is a common cause of endocrine failure such as islet lipotoxicity, the effect of obesity on the thyroid follicular cells has not been investigated.[br][bold]Methods;[/bold] We performed the phenotype analyses and observed the histopathological adiposity features using the thyroid glands from [italic]ob/ob[/italic] mice, [italic]db/db[/italic] mice and diet-induced obesity (DIO) mice.[br][bold]Results;[/bold] The histopathological analysis on male and female C57BL6 control mice fed on normal chow revealed characteristic brown and white adipose cells in the interfollicular area. Interestingly, genetically obese models, [italic]ob/ob [/italic]and [italic]db/db[/italic] mice showed striking increase of white adipose cells in the interfollicular area. Furthermore, Oil-red O and Bodipy staining clearly showed intracellular lipid droplet in the follicular cells of [italic]db/db[/italic] mice. The thyroid glands from the DIO mice that were fed the high fat diet for 6 weeks showed no histological changes. However, real-time RT-PCR showed that the expression of lipogenic genes such as SREBP1 and FAS is markedly increased in the thyroid glands of DIO mice, which was accompanied the endoplasmic reticulum (ER) swelling and increased UPR response demonstrated by electro-microscopic study and immunohistochemistry, respectively. Interestingly, the blood levels of thyroid hormone were lower in the DIO mice compared to the control mice fed normal chow although the DIO mice showed increased induction of thyroid specific genes such as TPO, TSHR and Tg.[br][bold]Conclusions; [/bold]Our observation suggested that the thyroid gland is one of target organs for ectopic fat deposition in obesity, which can induce the functional changes of thyroid follicular cells.[br][br]Sources of Research Support: This work was supported in part by the second phase of the Brain Korea 21 program of the Ministry of Education and by the Korea Healthcare technology R[amp]D Project, Ministry for Health, Welfare and Family Affairs, Republic of Korea (A100588). MHL, SEL and YSJ were supported by NRF/MEST (No. 2010-0005462).[br][br]Nothing to Disclose: MHL, JUL, MJC, MJR, SJK, YKK, KSK, MS, YSJ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 69 191 1444 SUN-417 PO39-02 Sunday 1241 2012


1238 ENDO12L_SUN-418 POSTER SESSION: Thyrotoxicosis, TSH Administration [amp] Thyroid Cell Biology (1:30 PM-3:30 PM) Influence of Thyroid Dysfunction on Serum Concentrations of Angiopoietin-Like Protein 6 and Fibroblast Growth Factor-21 Jung Ah Lim, Hoon Sung Choi, Hwa Young Ahn, Yenna Lee, Sung Hee Choi, Young Joo Park, Ka Hee Yi, Do Joon Park, Hak Chul Jang, Seong Yeon Kim Seoul National University College of Medicine, Seoul, Republic of Korea; Seoul National University Bundang Hospital, Seongnam, Republic of Korea; Boramae Municipal Hospital, Seoul, Republic of Korea Angiopoietin-like protein 6 (ANGPTL6) and fibroblast-growth factor 21 (FGF21) are novel metabolic regulators. Both of them modulate energy expenditure as well as glucose and lipid metabolism. Thyroid hormone can induce metabolic changes that are similar to those induced by ANGPTL6 and FGF21. We investigated whether circulating ANGPTL6 and FGF21 concentration are associated with thyroid hormone level in human. We measured the serum concentrations of ANGPTL6 and FGF21 and metabolic parameters in five groups of persons who exhibited overt hyperthyroidism (n=30), subclinical hyperthyroidism (n=30), euthyroidism (n=30), subclinical hypothyroidism (n=30) and overt hypothyroidism (n=30). Serum ANGPTL6 and FGF21 concentrations were significantly higher in patients with overt hypothyroidism than other persons. Women had higher serum levels of ANGPTL6 than men. ANGPTL6 concentration correlated positively with TSH, total cholesterol, AST and ALT in all subjects, and negatively with free T4. Multiple stepwise linear regression revealed that sex and free T4 were independent predictors of serum ANGPTL6 concentration. Serum FGF21 concentration correlated positively with triglyceride, and inversely with free T4. In cell culture studies, HepG2 cells treated with T3 in time dependent manner showed no significant difference in ANGPTL6 and FGF21 expression. Our results suggest that serum ANGPTL6 and FGF21 concentration are increased in patients with overt hypothyroidism. Free T4 level can be an independent predictor of serum ANGPTL6 level. However, thyroid hormone may have an indirect effect on the expression of ANGPTL6 and FGF21. Whether increased serum ANGPTL6 and FGF21 concentration in overt hypothyroidism are compensatory mechanism to increase an energy metabolism remains to be determined.[br][br]Nothing to Disclose: JAL, HSC, HYA, YL, SHC, YJP, KHY, DJP, HCJ, SYK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 793 191 1445 SUN-418 PO39-02 Sunday 1242 2012


1239 ENDO12L_SUN-419 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Effective Treatment of Hypothyroidism with Powdered Medicine and Its Involvement of Efflux Transporter BCRP in a Patient with Multidrug Resistance Tomohiko Yoshida, Mayumi Shoji, Seiichiro Higuchi, Ikki Sakuma, Hidekazu Nagano, Tomoko Takiguchi, Emi Imada, Naoko Hashimoto, Sawako Suzuki, Keiko Suyama, Hiromitsu Nakasa, Ichiro Tatsuno, Kotaro Yokote, Tomoaki Tanaka Chiba University Hospital, Chiba, Japan; Chiba University Hospital, Chiba, Japan; Toho University Sakura Medical Center, Sakura, Japan [bold]Introduction[/bold][br]Multidrug resistance (MDR) is characterized by a cross-resistance among structurally and mechanistically diverse agents, and is a serious complication during cancer chemotherapy. However, little is known about the association between MDR and endocrine diseases. Here, we report a case of MDR with severe primary hypothyroidism. Notably, whereas multiple oral tablets had little or no effect, the same ingredients were highly effective when they were intravenously administrated. Since it was previously reported that some patients responded to pulverized formulation of levothyroxine but not to tablet (1), we examined the effect of different types of drug formulations and determined whether the efflux transporter BCRP/ABCG2, which play a major role in MDR in cancers, is implicated in its pathophysiology.[br][bold]Case Reports[/bold][br]An 80-year-old woman with Hashimoto[apos]s thyroiditis, idiopathic thrombocytopenic purpura (ITP) and hypertension, exhibited resistance to the tablet formulations of levothyroxine, prednisolone, and antihypertensives. Poor oral bioavailability led us to treat by intravenous administration. Biweekly IV treatment of levothyroxine (300 [mu]g) was sufficient to maintain normal TSH and free T4 levels. Interestingly, consistent with the previous report, pulverized formulation of levothyroxine at a dose of 300 [mu]g per day was effective to maintain TSH and free T4 levels within the normal range, whereas despite using tablet formulation doses up to 600 [mu]g per day, patient[apos]s TSH levels remained very elevated (Average TSH level 67.8 [mu]IU/ml). Moreover, ITP and hypertension were also improved with powdered prednisone and antihypertensives. Since ATP-binding cassette (ABC) transporters are known to contribute MDR, we examined their gene expression profiles and functional analysis using peripheral blood mononuclear cells (PBMCs) and duodenal biopsy samples. Flow cytometric analysis of efflux transporter function revealed that drug excretion was accelerated in the patient[apos]s PBMCs and it was blocked by fumitremorgin C, a selective inhibitor of BCRP, in a dose dependent manner. Furthermore, among efflux transporters, [italic]BCRP[/italic] mRNA expressions were significantly increased in the patient[apos]s PBMCs and intestine.[br][bold]Conclusion[/bold][br]Our findings provide clinical and mechanistic insights into the poor oral bioavailability of MDR with Hashimoto[apos]s thyroiditis, and suggest a possible involvement of accelerated transporter function due to elevated expression of BCRP in the intestine.[br][br](1) Yamamoto T. Thyroid 2003; 13(12): 1177 -1181.[br][br]Sources of Research Support: Grant-in-Aid for Scientific Research (C) by Japan Society for the Promotion of Science 22590999.[br][br]Nothing to Disclose: TY, MS, SH, IS, HN, TT, EI, NH, SS, KS, HN, IT, KY, TT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1564 192 1446 SUN-419 PO56-02 Sunday 1243 2012


1240 ENDO12L_SUN-420 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Elevated TSH and Free Thyroxine: Important Differential Diagnoses Joshua D Maier, Sherry Ryan, Surender Arora LSUHSC-Shreveport, Shreveport, LA; Overton Brooks VA Medical Center, Shreveport, LA [bold]Background:[/bold] Many cases of incongruent TSH and thyroxine levels result from assay errors, interfering antibodies, or nonthyroidal illness[sup][1][/sup]. However, two rare conditions that cause both elevated TSH and free thyroxine, resistance to thyroid hormone (RTH) and TSH-secreting pituitary adenoma, are important to recognize due to differences in treatment [sup][2-7][/sup]. [bold]Clinical Cases: [/bold](a) A 52 year old male complained of chronic anxiety, tachycardia, sweating, and had been told of abnormal thyroid function tests several times previously. He appeared euthyroid with pulse 70 bpm, normal thyroid exam, and few other findings. His TSH was 2.87 [mu]IU/L (0.34-5.6), free T4 1.92 ng/dL (0.61-1.12), and free T3 3.89 pg/mL (2.5-3.9), with similar findings on serial determinations. T4 by dialysis was 2.9 ng/dL (0.8-1.7) and alpha subunit 0.4 ng/mL (0.09-0.76), molar ratio 1.32. Pituitary MRI was normal. He subsequently had several emergency department visits for palpitations. I-123 scan was homogeneous and symmetric. Percent uptakes were 15.2% at 4 hours (hrs) and 40.3% at 24 hrs. After 25 [mu]g of triiodothyronine TID for 10 days, suppression labs were TSH 0.06 [mu]IU/mL, free T4 1.35 ng/dL, free T3 10.6 pg/mL. I-123 suppression scan uptakes were 3.5% at 4 hrs and 7% at 24 hrs. This scenario is consistent with RTH. (b) A 42 year old male transferred the care of his hyperthyroidism to us. Three years earlier he was evaluated for headache. MRI revealed a 3.6 x 2.7cm sellar mass. He had craniotomy and biopsy, which disclosed a pituitary tumor. He was given external beam radiation. Four months later he noted heat intolerance and tachycardia. He had a goiter. TSH was 5.12 [mu]IU/mL (0.34-4.82), free T4 4.5 ng/dL (0.8-1.9). Thyroid scan was symmetrically homogenous with uptakes of 13.4% at 4 hrs and 65% at 24 hrs. He was placed on methimazole by his primary physician. At our evaluation, his TSH was 4.86 [mu]IU/mL (0.34-5.60)), free T4 1.99 ng/dL (0.61-1.12) and free T3 5.10 pg/mL (2.5-3.9), alpha subunit 11 ng/mL (0.05-0.53), molar ratio 22.63. MRI disclosed a 2.4 x 2.3cm sellar tumor. Octreoscan was positive at the sella. Re-analysis of the original biopsy was done, showing weak TSH staining. He began long-acting lanreotide and had repeat transsphenoidal surgery. A large residual remains on MRI. [bold]Conclusion:[/bold] The differential for persistently elevated TSH and free T4 is narrow. The alpha subunit level may help distinguish RTH from TSH-producing pituitary adenoma.[sup][2,3,4,6][/sup][br][br]1. Braverman LE and Utiger RD. Werner [amp] Ingbar[apos]s The Thyroid: A Fundamental [amp] Clinical Text. Lippincott Williams [amp] Wilkins, 2005. 2. Refetoff S and Dumitrescu AM. Syndromes of reduced sensitivity to thyroid hormone: genetic defects in hormone receptors, cell transporters and deiodination. Best Pract Res Clin Endocrinol Metab. 2007 Jun; 21(2):277-305. 3. Beck-Peccoz P [amp] Chatterjee VKK. The variable clinical phenotype in thyroid hormone resistance syndrome. Thyroid 1994; 4: 225-232. 4. Beck-Peccoz P, Brucker-Davis F, Persani L, Smallridge RC, Weintraub BD. Thyrotropin-Secreting Pituitary Tumors. Endocr Rev. 1996 Dec; 17(6): 610-638. 5. Beck-Peccoz P, and Persani L. Medical Management of Thyrotropin-Secreting Pituitary Adenomas. Pituitary 2002; 5(2):83-88. 6. Socin HV, Chanson P, Delemer B, Tabarin A, Rohmer V, Mockel J, Stevenaert A, Beckers A. The changing spectrum of TSH-secreting pituitary adenomas: diagnosis and management in 43 patients. Eur J Endocrinol. 2003 Apr;148(4):433-42. 7. Kienitz T, Quinkler M, Strasburger CJ, Ventz M. Long-term management in five cases of TSH-secreting pituitary adenomas: a single center study and review of the literature. Eur J Endocrinol. 2007 Jul;157(1):39-46.[br][br]Nothing to Disclose: JDM, SR, SA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2067 192 1447 SUN-420 PO56-02 Sunday 1244 2012


1241 ENDO12L_SUN-421 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Myxedema Madness: A Case of Misdiagnosed Psychosis Vanessa Arguello, Farah Hasan, Tahira Yasmeen University of Illinois-Advocate Christ Medical Center, Oak Lawn, IL Background: Myxedema madness refers to the common association of myxedema and serious psychiatric complaints. Myxedema madness can be the first sign or symptom observed in the severely hypothyroid patient and if not aggressively treated can lead to high mortality rates, 20% or higher, from myxedema coma.[br]Clinical Case: A 43-year-old woman with hypothyroidism, non-compliant with medications, who presented with a 5-day history of visual and auditory hallucinations. She described seeing deceased family members and hearing their voices or noises from appliances. Patient was initially misdiagnosed with narcotic psychosis from recent opiate use. The initial examination demonstrated advanced myxedema with puffy face, bilateral non-pitting edema, dry doughy skin, delayed deep tendon reflexes, hypertension, and hypoxia. Her mental status gradually worsened, resulting in deep coma necessitating mechanical ventilatory assistance. A clinical diagnosis of myxedema coma with myxedema psychosis was established and preemptive treatment was initiated with intravenous (IV) hydrocortisone then IV levothyroxine. Her hormone profile confirmed clinical hypothyroidism (elevated TSH and low FT4 and FT3): TSH 83.2 mcU/mL (reference range [RR] 0.350-5.0), FT4 0.70 ng/dL (RR 0.8-1.5), and FT3 1.3 pg/mL (RR 2.2-4.0). The reduction in FT4 and FT3 were less prominent at the time of measurement given patient had received 1-week treatment of levothyroxine 100 mcg/day. A normal am cortisol 30.7 mcg/dL (RR 6-23) confirmed intact pituitary-adrenal axis. After one day of treatment, her mental status dramatically improved and no longer required ventilatory assistance. Her psychosis resolved upon awakening from myxedema coma. Her myxedematous mental state and appearance continued to improve and the patient was discharged on oral levothryoxine 200 mcg/day. The precipitating factor for the myxedema psychosis was attributed to her recent opiate use. A one-month phone follow-up with patient confirmed complete resolution of auditory and visual hallucinations. In addition, she reported improvement in mental alertness, memory, bilateral feet edema, and a 14-lb weight loss.[br]Conclusion: Myxedema madness is rare and often overlooked by health care providers as a cause of organic psychosis. This case highlights psychosis as part of the myriad of clinical presentation for myxedema and stresses the importance of prompt identification to avoid other more serious myxedema sequelae.[br][br](1) Asher R. Myxoedematous madness. Br Med J. 1949 Sep 10;2(4627):555[ndash]562. (2) Heinrich TW, Grahm G. Hypothyroidism presenting as psychosis: myxoedema madness revisited. Prim Care Companion J Clin Psychiatry 2003; 5: 260[ndash]6.[br][br]Nothing to Disclose: VA, FH, TY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1168 192 1448 SUN-421 PO56-02 Sunday 1245 2012


1242 ENDO12L_SUN-422 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Graves Ophthalmopathy and Pretibial Myxedema with Primary Hypothyroidism Payal Satish Patel, Naifa Busaidy, Camilo Jimenez Baylor College of Medicine, Houston, TX; University of Texas MD Anderson Cancer Center, Houston, TX Introduction: Ophthalmopathy and pretibial myxedema are typically associated with primary hyperthyroidism. It is rare to see both of these entities in a patient with primary hypothyroidism.[br]Clinical Case: A 74-year-old woman with multiple myeloma and recently diagnosed Graves[apos] ophthalmopathy and pretibial myxedema was referred to endocrine clinic for thyroid evaluation. She began having eyelid erythema and edema and increased tearing in 2/2011. She evaluated by Ophthalmology and was diagnosed with bilateral proptosis (NOSPECS class IV) with Hertel measurements of 24mm bilaterally (upper limit of normal is 20mm). She denied any blurring, diplopia, or loss of vision and had full range of motion of extraocular muscles. A CT scan of the orbits showed enlargement of the superior, inferior, and medial rectus muscles bilaterally and excessive fat within the orbits, compatible with Graves[apos] ophthalmopathy.[br]She also had swelling and erythema circumscribing the distal one-third of the right lower leg that appeared in 1/2011. After topical hydrocortisone ointment failed to improve the rash, she was referred to dermatology. Biopsy of the erythematous area was performed at an outside facility with pathology consistent with myxedema.[br]Furthermore, patient reported a history of primary hypothyroidism for nine years. She was taking levothyroxine 100 mcg/day and denied any history of hyperthyroidism preceding the diagnosis of hypothyroidism. Laboratory tests revealed the following: TSH 0.90 uIU/mL (0.27-4.20), Free T4 1.57 ng/dL (0.93-1.70), TSI Index 6.7 ([lt]1.3), and TPO antibody 948 IU/mL ([lt]35).[br]Given normal thyroid function tests, she was continued on levothyroxine 100mcg/day. She is being considered for a course of steroids for the ophthalmopathy.[br]Conclusion: Although Graves[apos] ophthalmopathy has been reported in association with euthyroidism and primary hypothyroidism, literature review revealed only four previous cases of patients with both Graves[apos] ophthalmopathy and pretibial myxedema in the setting of primary hypothyroidism. Management of ophthalmopathy is the same regardless of the underlying primary thyroid disorder. Selenium can be considered for mild cases of ophthalmopathy, steroids for moderate eye disease, and orbital decompression for severe, sight-threatening eye disease.[br][br]Nothing to Disclose: PSP, NB, CJ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1131 192 1449 SUN-422 PO56-02 Sunday 1246 2012


1243 ENDO12L_SUN-423 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) A Rare Case of Otherwise Asymptomatic Hypothyroidism Presenting with Hypertensive Emergency and Cardiac Tamponade Shilpa A Chaudhari, Sunjit S Jaspal, Ashvin Butala, Alan S Sacerdote, Gul Bahtiyar Woodhull Medical Center, Brooklyn, NY; New York University School of Medicine, New York, NY; State University of New York (SUNY) Downstate Medical Center, Brooklyn, NY; St George[apos]s University School of Medicine, WI, Grenada Objective: To present a rare case of otherwise asymptomatic hypothyroidism presenting with hypertensive emergency and pericardial tamponade.[br]Patient, Methods and Case Report:[br]A 39 yo female presented to the Emergency Department (ED) complaining of chest pain, myalgia, decreased baseline tolerance, and swelling of the lower extremities. In the ED, her blood pressure (BP) was 202/117 mm Hg; apical heart rate was 75 bpm. On auscultation, distant, muffled heart sounds were heard, without murmur or gallop; no other features of hypothyroidism were noted. On palpation, a normally sized thyroid gland without palpable nodules was noted. ECG showed diffuse T wave flattening in the anterolateral leads. Shortly after admission, she developed increasing respiratory distress and required intubation. Her BP dropped to 80 mm Hg after intubation. Bedside echocardiogram was consistent with cardiac tamponade and diastolic collapse of right atrium and ventricle. Surgery was consulted and immediate pericardiocentesis was done. Analysis of pericardial fluid and other laboratory data ruled out all common causes except for hypothyroidism as a cause of cardiac tamponade. Serum TSH by chemiluminescence was 75.32 mIU/L (0.40-5.50), total serum cholesterol was 280 mg/dl, LDL calculated was 197 mg/dl.[br]Discussion:[br]Only a very few cases have been reported with hypertension as part of the presentation of cardiac tamponade in myxedema patients. This is a unique presentation in a patient who was generally asymptomatic with regard to hypothyroid patient. In hypothyroid patients, normal BP is maintained due to slow progression of pericardial distension allowing compensatory mechanisms to be activated. In our patient the BP remained elevated after pericardiocentesis. Possible mechanisms for hypertension could be increased serum catecholamine levels, enhanced nonadrenergic reactivity of vascular smooth muscle, and an increase in binding the alpha-1 adrenergic receptor as described by Brown et al (1).[br]Conclusions:[br]- In patients presenting with hypertensive emergencies and cardiac tamponade, it is crucial to check the thyroid function tests though the patient may not have any other signs or symptoms of hypothyroidism.[br]- Proper diagnosis and timely thyroid hormone replacement will improve the prognosis in such patients.[br][br]Nothing to Disclose: SAC, SSJ, AB, ASS, GB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 37 192 1450 SUN-423 PO56-02 Sunday 1247 2012


1244 ENDO12L_SUN-424 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) A Rare Complication of Hypothyroidism Ramya Embar Srinivasan, Susie Myer Estes University of Tennessee College of Medicine Chattanooga, Chattanooga, TN Objective:[br]To report a case of ascites and pericardial effusion as a rare complication of primary hypothyroidism.[br]Case History: The patient is a 43-year-old white female with a history of hypothyroidism off her thyroid replacement for a year presented with irregular periods, weight gain, hoarse voice, hair loss and abdominal distention with dyspnea. Physical exam significant for thin hair, dry skin, decreased reflexes, non-pitting edema in her extremities, distant heart sounds, dullness on abdominal percussion. Thyroid stimulating Hormone(TSH) level was 55 and free T4 was less than 0.4ng/dl. Four liters of straw colored fluid removed by paracentesis revealed exudative effusion with total protein 6gm/dl, total serum protein 7.1gm/dl, serum albumin 3.5gm/dl, ascitic fluid albumin 3.4gm/dl, serum ascites albumin gradient (SAAG)0.1gm/dl, fluid wbc 795cells/mm[sup]3[/sup], lymphocytes 58%. Further evaluation to exclude other causes of high protein ascites was negative including malignant cells, bacterial cultures, Acid fast bacilli stain and culture, urine protein for nephrotic range and antinuclear antibody. No evidence of heart failure on echocardiogram except for large pericardial effusion without tamponade. Ultrasound of the liver was negative as well as CT scan of the abdomen and pelvis except for large amount of ascites.[br]Discussion:[br]Pericardial effusion is seen in 3-6% of cases with hypothyroidism[sup]1[/sup]. Ascites caused by hypothyroidism occurs in [lt]4% of cases, so its diagnosis is often delayed and patients frequently receive unnecessary procedures such as liver biopsies and exploratory laparotomies[sup]5[/sup]. Review of literature reveals that analyses of ascitic fluid from patients in this condition usually shows high protein ([gt]2.5 g/dL), low white blood cell counts, with a high proportion of lymphocytes[sup]2,5[/sup]. Case reports of both high[sup]3[/sup] as well as low SAAG[sup]4[/sup] are documented. Our patient had a low SAAG, high protein and higher proportion of lymphocytes. Her TSH level was 55, literature review showed no correlation between TSH level and ascites. A consistent feature is the response to thyroid hormone replacement therapy, which has always led to resolution of the ascites. We opted for watchful waiting with thyroid replacement once we excluded the commonest causes of high protein ascites without making the patient undergo extensive invasive workup.[br]Conclusion: Myxedema ascites is rare but easy to treat; it should be in the differential diagnosis, especially if the ascites fluid has a high protein content.[br][br]1.Pericardial effusion in primary hypothyroidism. Kabadi UM et all, Am Heart Journal 1990 Dec 120(6 Pt 1):1393-5. PMID 2248183. 2.Ji JS, Chae HS, Cho YS, Kim HK, Kim SS, Kim CW, Lee CD, Lee BI, Choi H, Lee KM, Lee HK, Choi KY. J Korean Med Sci. 2006 Aug;21(4):761-4. Review. PMID: 16891828. 3.De Castro,F; Bonacini M;Walden J;Schubert,T.Myxedema Ascites: Report of Two cases and review of literature J Clinical Gastro 1991;13:411-4. 4.Otero Bedoyo J,Landeira G,Corino M Tamashiro A,Fassio E. Ascites due to hypothyroidism in a patient with alcoholic cirrhosis.Acta Gastroenterol Latinoam.2001;31:77-81. 5.Chiprut RO,Knudsen KB,Liebermann TR,Dyck WP Myxedema ascites Am J Dig Dis. 1976;21:807-808.[br][br]Nothing to Disclose: RES, SME 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 569 192 1451 SUN-424 PO56-02 Sunday 1248 2012


1245 ENDO12L_SUN-425 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Severe Hypothyroidism Presenting with Ventricular Syncope and New-Onset Gynecomastia with Levothyroxine Treatment Yew-Xin Teh, Ali Rizvi University of South Carolina School of Medicine, Columbia, SC [bold]Objective:[/bold] To describe ventricular torsades as the presenting manifestation of hypothyroidism, and new-onset gynecomastia after treatment with levothyroxine.[br][bold]Background:[/bold] Severe hypothyroidism may present with EKG abnormalities including bradycardia, low voltage, and heart block. Rarely, patients may manifest ventricular ectopy. Gynecomastia has been described in thyrotoxicosis but is unusual after treatment of hypothyroidism.[br][bold]Case Report:[/bold] A 34-yr-old previously healthy man with palpitations and syncope felt light-headed during an exercise stress test. EKG revealed prolonged QT interval, runs of ventricular tachycardia (VT), and torsades de pointes. Cardiac catheterization showed non-ischemic cardiomyopathy and EF of 35%. The patient complained of tiredness, bloating, and hair loss. Tests showed a TSH [gt] 400 uIIU/ml, free T4 0.11 ng/dl (0.8-1.8), and free T3 0.2 ng/dl (2.4-4.2). Levothyroxine (LT4) 25 mcg PO daily was started and the dose gradually titrated up on a weekly basis. Four months later he was clinically euthyroid. Tests showed TSH 0.23 uIIU/ml, free T4 1.44 ng/dl, normal EKG, and no ectopy. The patient complained of new bilateral breast enlargement and tenderness. Exam confirmed gynecomastia without any discrete mass or nipple discharge, and no testicular or scrotal masses. Hormonal work-up showed total testosterone (T) 380 ng/dl (241-827), free T 69.2pg/ml (74-242), estrogen 48 pg/ml (11-41), SHBG 39 nmol/l (13-71), LH 3.4 mIU/ml (1.5-9.3), prolactin 6.2 ng/ml (2-18) and undetectable HCG. The patient was reassured that the breast enlargement was benign and most likely secondary to hormonal alterations related to thyroid treatment.[br][bold]Discussion/Conclusion:[/bold] The case illustrates two unique aspects related to hypothyroidism and its treatment. A prolonged QT interval in myxedema can rarely predispose to life-threatening VT and torsades, manifesting with palpitations and syncope (1,2). This uncommon but potentially lethal cardiac complication of hypothyroidism is nevertheless completely reversible with treatment (3). The patient also developed new-onset gynecomastia with evidence of LT4 over-replacement, mild TSH suppression, reduced free T, and elevated estrogen. Although described classically in florid hyperthyroidism with resultant increase in SHBG levels and relative estrogen-testosterone ratio (4,5), the same underlying hormonal changes may account for transient breast tissue proliferation with over-correction of profound hypothyroidism (6).[br][br]1. Lim CH et al., Aust N Z J Med 1976;6:68[ndash]70. 2. Lehmann NH et al., Am J Cardiol 1997;79:963-5. 3. Schenck JB et al., Am J Med Sci 2006;331(3):154-156. 4. Tan YK et al., J R Coll Surg Edinb 2001;46(3):176-7. 5. Chan WB et al., Postgrad Med J 1999;75(882):229-31. 6. Ho HK et al., Ann Acad Med Singapore 1998;27(4):594-6.[br][br]Nothing to Disclose: Y-XT, AR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 21 192 1452 SUN-425 PO56-02 Sunday 1249 2012


1246 ENDO12L_SUN-426 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) An Uncommon Presentation of Hypothyroidism Vijay G Eranki, Alan Silverberg Saint Louis University School of Medicine, St Louis, MO Introduction[br]Hyperprolactinemia can occur in patients with primary hypothyroidism, but it is uncommon to see a patient with severe hypothyroidism present with only symptoms of galactorrhea and a pituitary mass.[br]Clinical case[br]A 35 year old woman presented to the clinic with complaints of galactorrhea and pituitary mass. She had two uncomplicated pregnancies and deliveries, the family and social history were non contributory. The patient nursed her infant for three years and was amenorrheic. Her periods did not resume even after she stopped nursing. A year after cessation of nursing, she noted nipple discharge. She had a pituitary MRI which showed a sellar mass with supra sellar extension 14x15x12 mm in size. The patient was evaluated by neurosurgery, and lab showed prolactin of 49.8 ng/mL (3.2-29.1). She was then referred to endocrinology.[br]In the endocrine clinic, the history was significant for energy loss, bilateral headaches and weight gain associated with constipation. The physical exam was normal including an assessment of the visual fields. However, the thyroid was noted to be diffusely enlarged and thick. Other labs done revealed TSH [gt]150 uIU/mL (0.35-5.5) and free T4 of 0.38 ng/dL (0.61-1.76) and she was started on levothyroxine 50 mcg daily.[br]On treatment with the levothyroxine the energy level improved, headaches, nipple discharge and the amenorrhea resolved. The follow up labs showed TSH 12.1 uIU/mL (0.35-5.5), total T4 5.1 mcg/dL (4.5-12.5), free T4 0.9 ng/dL (0.7-1.9), free T3 1.7 pg/mL (1.5-3.5), prolactin 21.2 ng/mL (3.2-29.1), IGF I 137 ng/mL (63-330) and anti TPO was [gt]1000 IU/mL (0-3.9). The levothyroxine dose was titrated up to achieve a TSH of 0.816 uIU/mL (0.27-4.67). A follow up MRI three months later showed resolution of the sellar mass and no significant supra sellar lesion was noted.[br]Discussion/Conclusion[br]The mechanism of the prolactin elevation in this setting is not completely clear. TSH should be measured in all patients presenting with galactorrhea and a suspected prolactinoma to rule out hypothyroidism, it is important not to presume it to be due to secondary hypothyroidism from tumor involvement of the pituitary.[br][br]Grubb MR, Chakeres D, Malarkey WB: Patients with primary hypothyroidism presenting as prolactinomas. Am J Med. 1987 Oct;83(4):765-9.[br][br]Nothing to Disclose: VGE, AS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 909 192 1453 SUN-426 PO56-02 Sunday 1250 2012


1247 ENDO12L_SUN-427 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Autoimmune Hypothyroidism Converted to Hyperthyroidism: Is It a Common Phenomenon? Saira Furqan, Najmul Islam Aga Khan University and Hospital, Karachi, Pakistan [bold]Introduction:[/bold][br]Graves[apos] disease and Hashimoto[apos]s thyroiditis are the two autoimmune spectrum of thyroid disease. Cases of conversion from hyperthyroidism to hypothyroidism have been reported but conversion from hypothyroidism to hyperthyroidism is very rare although reported. We report a case of hypothyroidism that converted to a hyperthyroid state needing treatment.[br][bold]Case report:[/bold][br]A 36 yrs old female presented with a 3 months history of Easy fatigability, cold intolerance, polymenorrhagia, constipation and weight gain in the beginning of 2005. On examination she had bradycardia and dry skin. The thyroid gland was palpable, mostly diffuse but some nodular feeling at upper pole of left lobe. Clinical suspicion of primary hypothyroidism was made and it was than confirmed by TSH of greater than 50 with FT4 value of less than 0.30 and positive thyroid antibody titre. Thyroxine was started at a dose of 100 mcg/day. Gradually the requirement of thyroxine decreased and from end of 2005 onwards she maintained her TSH within normal range on 50 mcg/day of thyroxine. In the beginning of 2008 the dose was further reduced to 25 mcg/day but again towards the end of 2009 thyroxine dose was increased to 50 mcg/day because of slightly increased TSH of 8.86. Slightly more than a year later in the beginning of 2011 she presented with weight loss of 3 kg with a feeling of anxiety and associated tremors of hands. TSH at this time was less than 0.005 with a FT4 of 2.4 confirming the state of thyrotoxicosis. Thyroxine was stopped and patient was observed intermittently over a period of 6 months. She remained clinically and biochemically hyperthyroid with a repeat TSH of [lt]0.005 and an FT4 of 2.66. Thyroid scintigraphy with technetium 99 showed done at this stage showed an increased homogenous tracer uptake. Finally she was started on Carbimazole in mid of 2011and remains on it till to date.[br][bold]Conclusion: [/bold][br]This case demonstrate that high index of suspicion should be there if a patient with primary hypothyroidism develop persistent symptoms of hyperthyroidism. Otherwise it can be missed easily considering it as an over replacement with thyroid hormone.[br][br]Nothing to Disclose: SF, NI 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1433 192 1454 SUN-427 PO56-02 Sunday 1251 2012


1248 ENDO12L_SUN-428 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Amiodarone-Induced Hyperthyroidism Occurring 10 Months after Amiodarone Discontinuation Uma Muthyala, Alan Silverberg Saint Louis University, Saint Louis, MO Amiodarone is a commonly used anti-arrhythmic drug which can cause thyroid dysfunction. Typically amiodarone-induced thyrotoxicosis (AIT) occurs within months of amiodarone initiation. We describe a case of AIT occurring 10 months after discontinuing the medication.[br]71 year old female with history of atrial fibrillation presented with acute-onset of palpitations and dyspnea. She was treated with amiodarone for several years in the past and was discontinued 10 months ago as she was in sinus rhythm. She denied getting any kind of imaging with contrast and was also noted to be allergic to iodinated contrast. Physical examination revealed an irregularly irregular heart rate of 90 bpm.Thyroid examination did not reveal goiter and there were no other signs of hyperthyroidism other than tachycardia. Thyroid function tests revealed hyperthyroidism with TSH [lt] 0.015 mIU/l, FT4 3.5 ng/dl and FT3 6.1 pg/ml. A thyroid uptake and scan revealed low uniform uptake of 2.5 % at 24 hours. There was no abnormal uptake in the pelvic region. Two weeks later, FT4 and FT3 had further increased. IL6, ESR were normal and urinary iodine was elevated at 1306 (Normal 28 - 544 mcg/l).Treatment was initiated with methimazole.[br]Iodine-induced hyperthyroidism, also known as Jod-Basedow disorder typically occurs in association with iodine-containing medications, high iodine containing food and CT-contrast. It typically occurs early on in association with iodine load. Our patient is unusual as she had normal thyroid functions while on amiodarone treatment and presented 10 months after discontinuation of amiodarone in the absence of other recognizable forms of iodine ingestion. It is important to recognize late onset AIT, months after discontinuation as amiodarone has longer half life. 24 hour urinary iodine may aid in the diagnosis if the suspicion is high.[br][br]Nothing to Disclose: UM, AS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 70 192 1455 SUN-428 PO56-02 Sunday 1252 2012


1249 ENDO12L_SUN-429 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) A Case of Lithium Treatment Preceding Onset of Graves Disease: Effects on Presentation and Radionuclide Studies Jason Marley, Richard Comi Dartmouth-Hitchcock Medical Center, Lebanon, NH Lithium was first used in the 1800s for mania and remains the gold standard treatment of bipolar disorder today. The interaction of lithium and the thyroid has been known for hundreds of years. It is known that lithium increases intrathyroidal iodine and inhibits release of T4 and T3 and lithium therapy is commonly associated with hypothyroidism, less frequently associated with thyrotoxicosis, and rarely associated with Graves[apos] disease. A recent retrospective study of patients treated with lithium revealed an incidence of Graves[apos] 2-3 times greater than the general population. We present a patient who was diagnosed with Graves[apos] shortly after a significant decrease in her lithium doses, and whose thyroid scan and uptake suggest an effect of lithium that may have played into her presentation.[br]A 60 year-old female with bipolar disorder treated with lithium presented with worsening tremor, weakness, and unintentional 40 lb weight loss. Due to worsening tremor, her daily lithium dose had been decreased from 1200 mg to 300 mg several weeks prior to presentation. Physical exam revealed a thin middle aged female with proptosis and a coarse resting tremor. Labs revealed a lithium level of 0.96 mmol/L (therapeutic range: 0.6-1.2), TSH of [lt]0.01 mcIU/mL, T4 of 12.9 mcg/dL, T3 of 264 ng/dL, thyroid peroxidase antibody of 2990 IU/mL ([lt]34), and thyroid stimulating immunoglobin of 6.2 TSI ([lt]1.3). Thyroid uptake scan showed homeogeneous uptake: 9% at 4h, 11% at 24h. The patient was treated with PTU and there was subsequent improvement in free T4 and T3, as well as marked improvement in symptoms. She was discharged on methimazole, propanolol and lithium.[br]Lithium is known to inhibit thyroid hormone release and interfere with the egress of iodine from the thyroid gland, as well as having an association with thyroid autoimmunity. We speculate in this case that lithium therapy ameliorated symptoms and signs of hyperthyroidism and the rapid reduction in lithium therapy resulted in the emergence of clinical hyperthyroidism. The relatively low iodine uptake may have been the result of prior lithium therapy which traps iodine in the gland, reducing the entry of radioactive iodine into the intra-thyroidal iodine pool.[br][br]Nothing to Disclose: JM, RC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1644 192 1456 SUN-429 PO56-02 Sunday 1253 2012


1758 ENDO12L_MON-173 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Unexpected Hypoglycemia and Hyperglycemia Due to Insulin Antibody Developed in a Type 2 Diabetic Treated with Insulin Elaine YN Cheung, Grace YW Kam, Grace PS Hui, CS Hung, Flora CW Ho, MW Tsang United Christian Hospital, Hong Kong, Hong Kong Introduction: Occasional type 2 diabetic (T2DM) patients on insulin develop insulin antibody, leading to unexpected hypoglycemia and hyperglycemia.[br]Case history: A 72-year-old gentleman, with 10 years history of T2DM, was treated with oral hypoglycemic agents (OHA) until 2011. His T2DM is complicated by retinopathy, neuropathy and nephropathy with creatinine level up to 150umol/l. He developed OHA failure and was put on protaphane (PN) injection twice daily since April 2011 with no OHA. His HBA1c level came down from [gt]17% to 7.4% with pre meal hemoglucostix (h[apos]stix) around 4.5-13 mmol/l. However since September 2011, he developed fasting hypoglycemia. Because of this, the dosage of his nocte PN was reduced and finally stopped. He was kept on PN 18units on morning. He was advised to take regular bedtime snack and frequent small meals composing of carbohydrate with low glycemic index to reduce postprandial hyperglycemia and thus stimulus to insulin secretion. His fasting hypoglycemia persisted with h[apos]stix down to less than 3mmol/l with severe postprandial hyperglycemia up to 20mmol/l. PN was stopped and he was put on [alpha]-glucosidase inhibitor thrice daily. Insulin level checked during fasting hyperglycemia showed to be [gt]1000 uIU/ml. Insulin antibody level came back to be above 50U/ml. He was started on Prednisolone 30mg daily. There is no further episode of hypoglycemia but we are still up titrating his daytime insulin to attain good glycemia control.[br]DISCUSSION[br]With the usage of purified human insulin, the prevalence of insulin antibodies in insulin-treated patients have decreased to [lt] 50%. Such antibodies usually do not affect overall metabolic control, insulin dosage or leads to hypoglycemia. However in occasional patients like ours, development of antibodies can lead to change in insulin pharmacokinetics, resulting in high postprandial blood glucose levels and delayed hypoglycemia. This is due to the insulin-antibody interaction leading to lower initial rate of rise, delayed peak and longer time for return of plasma free insulin level to baseline. The antibody level checked during hypoglycemia was sky high, suggesting of dissociation of a large amount of insulin bound to antibodies at that juncture.[br]CONCLUSION[br]We report here a patient who developed insulin antibodies 5 months after start of PN leading to high postprandial blood glucose levels and fasting hypoglycemia. Fasting hypoglycemia was successfully treated with steroid.[br][br]Nothing to Disclose: EYNC, GYWK, GPSH, CSH, FCWH, MWT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2371 299 2094 MON-173 PO47-01 Monday 1762 2012


1759 ENDO12L_MON-174 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Is There an Increased Risk of Arthralgias in Specific Populations Using DPP-IV Inhibitors? Tira Chaicha-Brom, Tahira Yasmeen University of Illinois of Chicago, Chicago, IL; Advocate Christ Medical Center, Oak Lawn, IL This is a case of a 48yo Indian physician with a long standing history of type II diabetes mellitus, well controlled on metformin, who developed severe arthralgias with the use of DPP-IV (dipeptidyl peptidase-4) inhibitors. His Hgba1c was typically under control, less than 6.5%. However, his fasting blood sugars were becoming consistently elevated, so he chose to add Januvia (sitaglipitin) to his regimen. He did well on the medication for two weeks until he developed pain and erythema of his metacarpal joints. The arthralgias worsened and he developed morning stiffness; however, a rheumatological work-up was unrevealing for rheumatoid arthritis or other connective tissue disease. His ESR (erythrocyte sedimentation rate) was also low. The arthralgias became debilitating to the point that the patient contemplated disability due to the severe pain in his hands that limited his work. He even had difficulty walking up stairs and driving. However, his Hgba1c improved to 6.1%. Six weeks after starting Januvia, he developed a rash. He discontinued the medication and within 48 hours the rash resolved. By six weeks he had a marked improvement in the arthralgias. He then tried other DPP-IV inhibitors which again precipitated arthralgias.[br]There are two reported cases in the Japanese population that have demonstrated a causal association between the use of DPP-IV inhibitors and the development of remitting seronegative symmetrical synovitis (1). Although the cause-result relationship was not definitive, the authors argue that the temporal relationship of when the medication was initiated is highly suggestive. Our patient differs in that he is younger and with the trial of several other DPP-IV inhibitors medications it precipitated the same debilitating symptoms. Furthermore, his ESR was low. One proposed mechanism is that perhaps Asians or South Asians may have different levels of expressivity of the DPP-IV enzyme. Rat models have shown to have a decrease in CRP and an increase in TNF-alpha with the use of sitagliptin (2). Musculoskeletal pain has been noted with the use of DPP-IV (6). With the ever growing Asian population and the increased prevalence of diabetes within the Asian population, it is paramount to have medical therapy that will be well tolerated to prevent the complication of diabetes.[br][br]1) Yamauchi K, Sato Y, Yamashita K. RS3PE in Association with dipeptidyl peptidase-4 inhibitor: report of two cases. Diabetes Care 2012; 35:e7. 2) Ferreira L, et al. Effects of Sitagliptin Treatment on Dysmetabolism, Inflammation, and Oxidative Stress in an Animal Model of Type 2 Diabetes (ZDF Rat). Mediators of Inflammation 2010 (2010). 3) www.tradjenta.com. 4) www.januvia.com. 5) www.onglyza.com. 6) Richar, K. et al. Tolerability of Dipeptidyl Peptidase-4 Inhibitors: A Review. Clinical Therapeutics 2011 (33):11.[br][br]Nothing to Disclose: TC-B, TY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1794 299 2095 MON-174 PO47-01 Monday 1763 2012


1760 ENDO12L_MON-175 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Combination Therapy with Sitagliptin, Pioglitazone and Lansoprazole To Preserve B Cell in New Onset Type 1 Diabetes: Case Control Series Saad Sakkal Metabolic Care Center, Aleppo, Syrian Arab Republic OBJECTIVE: to determine if oral agents with B cell preservation effect,used to prevent DM, could decrease insulin dose or improve glucose control compared with usual care (Insulin) among new type1 diabetes patients[br]RESEARCH DESIGN AND METHODS: A randomized controlled,proof of concept, trial of new onset Type 1 diabetic patients with Clinical polyuria, weight loss, blood glucose [gt]400 mg/dl, hemoglobin A1C (HbA1c) [gt]10.0%, Insulin level less than 8,C peptide less than 2ng/dl.[br]Intervention patients received Sitagliptin 50 mg, lansoprazole 30 mg, and Pioglitazone 30 mg daily(n = 4),in addition to small doses of prandial insulin(4-6 units)as needed, versus usual care with insulin only given as three divided mixed doses 10-14 (n = 10) over a 1-year period.[br]The primary outcome is the percentage of patients achieving simultaneous control of glycaemia (FBS, 2HPP, HbA1c [lt]8.0%), clinical symptoms and signs (polyuria, weight regain) at 1 year. Secondary outcomes include improvements within each individual component of the composite primary outcome, daily Insulin dose, weight and hypoglycemia. Differences between groups were analyzed using t tests, Pearson [chi]2 tests.[br]RESULTS: patients age (10-18),(1M/3F.Vs.control3M/7F). all seen within 12 weeks of Symptoms onset.[br]At baseline: FBS was 511 in the interventional group and 461 in the insulin only group, HgA1C was 12.5% vs. 11.4, Insulin serum level was 7.3 Vs. 6,C peptide was 0.61 and 1.0,Insulin dose 32 and 29 was needed at onset, all were non-significant.AGAB were 5.6 and 11.7.[br]At six month a greater number of individuals assigned to oral B preservation Rx achieved the primary study outcome of having all three outcome measures under control (75% intervention Vs 60% usual care Insulin only group,P [lt] 0.01). In addition, a greater number of individuals assigned to the intervention group achieved the individual treatment goals of HbA1c [lt]8.0% (75% vs. 60%) and FBS/2HPP (75% vs. 50%), decrease Insulin dose (32 vs. 18), hypoglycemia (4 vs. 6)all with P value of[gt]0.01, but no difference noted for Polyuria (nocturnal and daytime:1/3), HgA1C (7.7 vs. 7.9) or weight (2 kg vs. 1.5). It is still too early to tell about longer term results.[br]CONCLUSIONS: In patients with new onset Type 1diabetes, oral beta cell preserving therapy with Sitagliptin,Pioglitazone and Lansoprazole can effectively improve control of Glycaemia and clinical status at six month with decreased insulin dose, and hypoglycemia. Long term data are needed.[br][br]Nothing to Disclose: SS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 245 299 2096 MON-175 PO47-01 Monday 1764 2012


1761 ENDO12L_MON-176 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Reversal of Abnormal Bladder Cancer FISH Assay Following Discontinuation of Thiazolidinedione and Continuation of Metformin Treatment Alan Scott Sacerdote, Gul Bahtiyar, Reema Batra Woodhull Medical [amp] Mental Health Center, Brooklyn, NY; State University of New York (SUNY) Downstate Medical Center, Brooklyn, NY; New York University School of Medicine, New York, NY; St George[apos]s University School of Medicine, St George[apos]s, Grenada Both thiazolidinediones, rosiglitazone and pioglitazone have been associated with a small, but significantly increased risk of bladder carcinoma. Recently, metformin use has been associated with a decreased risk of a variety of cancers.[br]Our patient, a 61 year old male, with Type 2 diabetes, metabolic syndrome, coronary artery disease, cerebrovascular disease, and peripheral arterial disease had been taking a combination of 4 mg rosiglitazone/1000 mg metformin twice daily for [ge] 5 years. On 01/22/2010 this combination was replaced with pioglitazone 45 mg daily plus metformin 1000 mg twice daily to improve his lipid profile. In 02/2010, the patient, who was taking enteric coated aspirin 81 mg and clopidogrel 75 mg daily following stent placement, presented with painless gross hematuria. Rectal examination and PSA by chemiluminescence were normal. Urine culture and cystoscopy were negative. Pelvic CT with contrast revealed possible mild thickening of the anterior bladder wall, possibly artifactual due to poor bladder distention. The Vysis UroVysion Bladder Cancer assay was performed on the patient[apos]s voided urine on 02-18-2010 by fluorescence in situ hybridization (FISH), which revealed 26 cells that were tetraploid or nearly tetraploid for chromosomes 2,7,17, and the 9p21 locus (normal [lt]10). This finding is consistent with either bladder carcinoma or [ldquo]umbrella[rdquo] cells-cells in the G2 phase of the cell cycle. A second FISH assay revealed similar findings. Pioglitazone was stopped on 03/03/2010. Follow-up cystoscopies remained negative. Repeat FISH assay performed on 09/08/2010 was normal.[br]We conclude that: 1) pre-cancerous/early cancerous changes in bladder epithelial cells associated with thiazolidinedione (TZD) use, in the absence of microscopic tumor, may be reversible when the TZD is stopped; 2) metformin may play an as yet undefined role in delaying formation of macroscopic bladder tumors and/or in reversing abnormal bladder cytology via the activation of AMP kinase and inhibition mTOR signaling.[br][br]Nothing to Disclose: ASS, GB, RB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 9 299 2097 MON-176 PO47-01 Monday 1765 2012


1762 ENDO12L_MON-177 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Recognizing Nonketotic Hyperglycemia Associated with Neurologic Symptoms Brandy Ma, Sanjay Navin Mediwala Baylor College of Medicine, Houston, TX; Michael E DeBakey VA Medical Center, Houston, TX Introduction: The association of nonketotic hyperglycemia (NKH) and seizure activity has been described but not adequately recognized in clinical practice.[br]Objectives: 1) Present a case of NKH associated with subclinical seizures and visual phenomena 2) discuss proposed mechanisms 3) review prior case reports[br]Clinical Case: A 61 yo male presented with a severe retro-orbital and right temporal headache with deteriorating visual acuity and visual hallucinations. The patient had a history of severe aortic stenosis and aortic insufficiency with valve replacement and aneurysm repair one-year prior, complicated by a posterior cerebral artery stroke. On admission, the patient had a blood glucose level of 605 mg/d, serum osmolality of 309mOsm/kg (n 280-301), and HbA1c of 14.2% (n 4.2-5.8). CT and MRI showed evidence of the prior right occipital infarct with encephalomalacia but no acute intracranial abnormalities or signs of occlusion or thrombosis. Temporal arteritis was suspected and prednisone treatment was initiated but later terminated when biopsy results excluded vasculitis. A subsequent EEG captured a focal subclinical seizure arising from and restricted to the right posterior quadrant. Therapy was initiated with valproic acid. Glucose levels were difficult to control and insulin therapy was titrated up to 65 units detemir and 35 units aspart with meals to maintain glucose levels between 120-150 mg/dL. The patient was discharged four days after the initiation of valproic acid treatment with adequate control of blood glucose levels and full resolution of visual symptoms. A 5-month follow-up in neurology, ophthalmology, and endocrine clinics showed good control of his diabetes (HbA1c 5.9%) and no recurrence of visual phenomena, residual visual field defects, or episodes of NKH.[br]Clinical Lessons: NKH presenting with seizure activity has been recognized in various case reports; however, the pathology remains unclear. Proposed mechanisms include decreased GABA production leading to lowering of the seizure threshold (1) and possible disruption of the blood-brain barrier (2). Preexisting or acute focal lesions may also predispose hyperglycemic patients to seizure activity.[br]Conclusion: Focal seizures are a known manifestation of NKH, and with most neurological symptoms readily controlled with the regulation of glucose levels, recognition of this association would expedite treatment and prevent the diagnostic dilemmas often associated with such cases.[br][br](1) Lavin PJ. Neurology, 2005 Aug; 65(4):616-9. (2) Kim DK et al. The Neurologist, 2011 May; 17(3): 164-6.[br][br]Nothing to Disclose: BM, SNM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 311 299 2098 MON-177 PO47-01 Monday 1766 2012


1763 ENDO12L_MON-178 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Poorly Controlled Diabetes: Consider Pheochromocytoma? Priyanka C Iyer, Artit Silpasuvan JHU/Sinai Hospital Internal Medicine Residency Program, Baltimore, MD Pheochromocytoma is often evaluated as a cause of secondary hypertension. However, pheochromocytoma is rarely on the differential diagnosis for poorly controlled diabetes. Catecholamines can cause hyperglycemia through stimulation of adrenoreceptors, through modulation of insulin and glucagon release and by stimulating hepatic glucose output and inhibiting skeletal muscle glucose uptake. We present the case of an insulin resistant patient who no longer required insulin days after resection of pheochromocytoma.[br]A 61 year old Caucasian man with a past medical history of poorly controlled hypertension, diabetes mellitus, and coronary artery disease, presented to the ER with episodic dyspnea, chest pain and diaphoresis. He was incidentally found to have an 11 cm adrenal mass during a CT scan done for chest pain evaluation. Biochemical workup revealed markedly elevated plasma free normetanephrines (5857pg/ml, n[lt]145), Metanephrines(8059pg/ml, n[lt]62pg/ml), urinary Metanephrines at 13988ug/L and elevated 24 hour normetanephrines (9608 ug/24 hours n: 110-1050). While admitted, his BP was 224/112 during symptomatic episodes. While being given over 160 total units of insulin daily, his fingersticks in the hospital ranged from 212 to 318 mg/dL. His Hba1c as an outpatient was 9.8% on this dose of insulin. Following 2 weeks pretreatment with phenoxybenzamine and beta-blockers, the patient underwent surgical right adrenalectomy. His post-op sugars ranged from 76-185 on only sliding scale insulin, when he would receive only 3 units daily. His BP was maintained without need for pressors. He was discharged off of all blood pressure medications as well as off all insulin. His 24 hour urine metanephrines collected 9 days post-operation were in the normal range. Pathology confirmed the adrenal mass to be a pheochromocytoma. On follow up one month later, he was noted to have a BP of 112/64 on only 5 mg of lisinopril. His fingerstick log showed all post meal sugars [lt]140 on only 850 mg of metformin once daily.[br]Conclusion: Biochemical testing for pheochromocytoma may be considered in patients with unexplained insulin resistance. Insulin dosages should be decreased rapidly post-operatively as insulin requirements drop dramatically.[br][br]Nothing to Disclose: PCI, AS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1867 299 2099 MON-178 PO47-01 Monday 1767 2012


1764 ENDO12L_MON-179 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Newly Developing Diabetes Mellitus after Roux-en-Y Gastric Bypass (RYGB) Lucie Favre, Luc Portmann, Fulgencio Gomez, Vittorio Giusti Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland [italic]Introduction[/italic]: Gastric Bypass (RYGB) has been shown to be the most efficient treatment for obese patients with type 2 diabetes mellitus (T2DM). RYGB improves glucose homeostasis through a variety of mechanisms including caloric restriction, stimulation of the entero-insular axis and diversion of nutrients into the distal gut. Thus, paradoxical onset or relapse of diabetes mellitus (DM) after RYGB should raise the suspicion of a different, new condition. We describe here two such cases.[br][italic]Case 1[/italic]: A 52 year-old obese (BMI 45.5 kg/m[sup]2[/sup]), hypertensive woman with normal glucose tolerance underwent RYGB. After an initial loss of 33 kg, her body weight stabilized. Three years later, she gained 6 kg over a short period, hypertension became resistant and diabetes was diagnosed (FBG 131 mg/dl, HbA1c 6.6%). On examination she had facial plethora, hirsutism, skin bruises and high blood pressure. Investigations showed elevated ACTH and cortisol, a large pituitary mass extending into the sphenoidal sinus, and a significant central-to periphery ACTH gradient on BIPSS. Transsphenoidal surgical removal of a pituitary adenoma, Ki67 [lt]1%, strongly expressing ACTH and GH, resulted in rapid resolution of Cushing[apos]s disease, secondary DM, and resistant hypertension.[br][italic]Case 2[/italic]: A 28 year-old obese (BMI 51 kg/m[sup]2[/sup]), hypertensive woman with dyslipidemia and T2DM, on metformin for 6 months, underwent a RYGB. She lost 50 Kg in 18 months (BMI 32 kg/m[sup]2[/sup]) and antihypertensive drugs and metformin were discontinued. At 3 yrs DM recurred (HbA1c 7.2%) and metformin was restarted. At 5 yrs she presented with a 10 kg weight loss due to decompensated DM. Strongly positive anti-GAD antibodies (1[apos]390[apos]800 IU/ml, N [lt] 10) led to the diagnosis of autoimmune DM (LADA type). Glucose control rapidly improved with insulin treatment.[br][italic]Discussion[/italic]: Although the incidence of T2DM has been described in 1% and 7% of patients respectively 2 and 10 years after bariatric surgery (especially gastric banding), no relevant data exist concerning RYGB and this has not been our experience. Such differences may depend on patients[apos] selection or rely on the surgical setting. A literature survey has shown no evidence linking bariatric surgery to the onset of pituitary tumors or autoimmune DM, and the association observed in these 2 cases was coincidental. Therefore, newly developing DM after RYGB should not be considered systematically as recurrence of T2DM but must lead to search for an alternative etiology of DM.[br][br]Nothing to Disclose: LF, LP, FG, VG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 934 299 2100 MON-179 PO47-01 Monday 1768 2012


1765 ENDO12L_MON-180 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) The Hyperinsulinism/Hyperammonemia Syndrome Complicated by Gestational Diabetes Mellitus: Treatment Challenges Ramona Dadu, Sonali Thosani, Ramaswami Nalini Baylor College of Medicine, Houston, TX [bold]Background: [/bold]Hyperinsulinism/hyperammonemia (HI/HA) syndrome is caused by gain of function mutations in the mitochondrial enzyme glutamate dehydrogenase (GDH) encoded by GLUD1 gene, which oxidizes glutamate to alpha-ketoglutarate and is a potential regulator of insulin secretion in pancreatic beta cells. Patients with HI/HA syndrome have both fasting and protein-sensitive hypoglycemia combined with asymptomatic elevations of plasma ammonia and generalized seizures. There are no reports of gestational DM in the setting of HI/HA syndrome so far.[br][bold]Case presentation:[/bold] A 25 year old woman G1P0 with history of HI/HA syndrome diagnosed at age 1 due to intractable seizures presented to us at 26 weeks gestation. Prior to pregnancy, she was doing well on diazoxide and cornstarch without recurrent hypoglycemia. During pregnancy, she stopped diazoxide but continued to take cornstarch. She had an episode of hypoglycemic seizure after a prolonged fast which resolved with glucagon administration. Family history was positive for type 2 DM in her maternal aunt, but negative for HI/HA syndrome. Physical examination was significant for an obese woman (BMI-38.3kg/m[sup]2[/sup]) with acanthosis nigricans on her neck. She had 25 pounds weight gain and developed hyperglycemia (fasting blood glucose of 112-175 mg/dL and 2-hour postprandial blood glucose of 107-187 mg/dL). Oral glucose tolerance test (100 gm) was consistent with gestational DM at 26 weeks (serum glucose: fasting- 114 mg/dL, 1 hour -197 mg/dL, 2 hour -176 mg/dL, and 3 hour -138 mg/dL). Hemoglobin A1c was 5.8% at 26 weeks and 6.9% at 36 weeks. Serum ammonia level was elevated during pregnancy (43-142 micromol/L, reference range: 11-32 micromol/L). Treatment included life style modifications and reduction in cornstarch amount with each meal. She was also advised to avoid protein-rich meals. These nutritional interventions resulted in improvement of glycemic control without significant hypoglycemia. She underwent cesarean section at 39 weeks gestation due to failure to progress and non-reassuring fetal heart tones. The baby weighed 8 lbs and 13 oz with apgar score of 9 and is doing well.[br][bold]Conclusion: [/bold]There is no data to guide treatment of gestational DM in patients with HI/HA syndrome. Once they develop insulinopenia/insulin resistance, they probably respond like all patients with type 2 DM. Treatment options include life style modifications and addition of insulin while monitoring carefully for hypoglycemia.[br][br]Nothing to Disclose: RD, ST, RN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1135 299 2101 MON-180 PO47-01 Monday 1769 2012


1766 ENDO12L_MON-181 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Autoimmune Dysglycemia: The Pendulum Swings Hope Torregosa, Sathya Krishnasamy, Stephen J Winters University of Louisville, Louisville, KY Insulin autoimmune syndrome and Type B insulin resistance are rare but notable causes of hypoglycemia. Each is characterized by the presence of hypoglycemia with elevated insulin levels, but can be differentiated by antibodies to either insulin or the insulin receptor. Antibodies either bind and release insulin, or serve as agonists or antagonists to insulin receptor signaling. We present a case in which both extreme hypoglycemia and hyperglycemia occurred.[br]Recurrent symptomatic hypoglycemia was noted in a 30-year old African American woman who was hospitalized for left hip replacement for avascular necrosis. She had systemic lupus erythematosus, lupus nephritis requiring hemodialysis after two failed kidney transplants, antiphospholipid antibody syndrome and heparin-induced thrombocytopenia. The patient was on maintenance prednisone dose of 10 mg and 5 mg on alternate days. Two months prior, her immunosuppressive agents sirolimus and mycophenolate mofetil were tapered and discontinued. When the fasting glucose was 40 mg/dL, the insulin level was [gt]1500 mIU/mL, with similar results a few days later. Insulin antibody level was elevated at 0.8 U/mL (normal [lt]0.4 U/mL), as were C-peptide at 36.6 ng/mL (0.8-3.20 ng/mL) and pro-insulin at 651 pmol/L ([le] 18.8 pmol/L). Computed tomography scan did not identify any pancreatic tumors. She was diagnosed with autoimmune hypoglycemia. She was discharged but shortly rehospitalized for sepsis and a flare of lupus arthritis requiring intravenous methylprednisolone. Her glucose level rose to [gt]900 mg/dL. Insulin level was [gt]1500 mIU/mL. C-peptide and proinsulin levels were elevated at 35.80 ng/mL and 427 pmol/L respectively, while the insulin antibody level was within the normal range. She was treated initially with insulin infusion and then U500 concentrated regular insulin at a total daily dose of 450 units, which was required to maintain euglycemia at a prednisone dose of 15 mg daily. After discharge, she slowly required less insulin which was ultimately discontinued.[br]Although autoimmunity to insulin or the insulin receptor is rare, recognition of this disease in predisposed individuals with hypoglycemia is necessary so that prompt diagnosis is made and appropriate management is applied. This case provided us with the full spectrum of hypoglycemia to hyperglycemia that can characterize either condition.[br][br]Nothing to Disclose: HT, SK, SJW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1866 299 2102 MON-181 PO47-01 Monday 1770 2012


1767 ENDO12L_MON-182 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Steroid Cell Tumor of the Ovary Presenting with Virilization and Uncontrolled Diabetes in a Caucasian Female Vanessa Escobar Barboza, Annis Marney University of Vermont, Burlington, VT Introduction[br]Ovarian sex cord stromal tumors are rare(5-8% of all primary ovarian neoplasms, with subtype steroid cell tumors representing only 0.1-0.5%. Hyperandrogenism is a common clinical manifestation that leads to an early diagnosis. [br]There are conflicting data about whether testosterone has a sensitizing effect on glucose metabolism in certain ethnic groups(African Americans. [br]We present a case of a Caucasian woman with an ovarian testosterone-producing tumor presenting with virilization and uncontrolled diabetes mellitus(DM).[br]Case Presentation[br]A 70 yo female presented with a 2 year history of increased facial hair, male pattern baldness, fatigue, and 15 pound weight loss over 3 months. Her past medical history is significant for Type 2 DM which had acutely worsened over the preceding 4 months with a hemoglobin A1c (HbA1c) 12.0% up from 7% the year before, coronary artery disease, hypertension, and hyperlipidemia. Her physical exam revealed obesity, thin hair on the top of her head, and facial hair whiskers. Abdominal exam revealed right lower quadrant tenderness to palpation. She had no clitoromegaly. Her free testosterone was 22.1 mg/dl (0.1-1.5), total testosterone 1198 mg/dl (14-76), sex hormone binding globulin 54.9 mg/dl (18-114), FSH 9.5 mg/dl, Estradiol 42 mg/dl, DHEA sulfate 85 mg/dl (0-157), cortisol 8 mg/dl, ACTH 13 mg/dl, 17 OH progesterone 258 mg/dl. A transvaginal US reported a mass within the endometrial cavity and an enlarged left ovary. An Abdominal CT reported an enlarged left ovary with a 2.9 cm solid mass, and normal adrenal glands. She underwent bilateral salpingoopherectomy a few weeks later. Her surgical pathology revealed a steroid ovarian cell tumor. A few weeks after, her free testosterone levels decreased to 25 mg/dl (14-76). In 2 months later, her hemoglobin A1c (HbA1c) decreased to 6.2%(from 12%). The only therapeutic modification during this time was the discontinuation of pioglitazone, and and the initiation of sitagliptin. At 6 months after surgery, her HbA1c was 7.3%, and she had gained 15 pounds since her surgery.[br]Conclusions[br]This case demonstrates the improvement in diabetes control after resolution of hyperandrogenism in a Caucasian female. The relationship between testosterone levels and insulin resistance/sensitivity is not well understood. As opposed to previous reports, this case shows that resolution of hyperandrogenism could potentially increase insulin sensitivity and improve glycemic control.[br][br]Nothing to Disclose: VEB, AM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2160 299 2103 MON-182 PO47-01 Monday 1771 2012


1768 ENDO12L_MON-183 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Recurrent, Concomitant Attacks of Gastroparesis and Sixth Nerve Palsy in a Woman with Type 1 Diabetes Sudhir Shah, Gul Bahtiyar, Harry Winters, Dmitriy Grinshpun, Alan Sacerdote, Peter Tse Woodhull Medical Center, Brooklyn, NY; State University of New York (SUNY) Downstate Medical Center, Brooklyn, NY; New York University School of Medicine, New York, NY; St George[apos]s University School of Medicine, St George[apos]s, Grenada Although neuropathy affects one-fifth of diabetics, recurrent episodic concomitant attacks of gastroparesis and sixth nerve palsy are rare.[br]Our patient, a 57 year old woman, had three distinct instances when she had simultaneous presentations of gastroparesis presenting with early satiety, frequent eructation, acid reflux, nausea and vomiting, as well as diplopia due to right sixth cranial nerve palsy. She had a history of type 1 Diabetes Mellitus (DM) for 45 years and been on insulin since diagnosis but her glucose control initially had been suboptimal. The diagnosis of gastroparesis was confirmed on the first presentation with single isotope nuclear gastric emptying time with administration of 99Tc sulfur colloid with a scrambled egg test meal for solids. T1/2 for solids was prolonged at [gt]150 min (normal:90-120 min). The initial impression of mononeuritis as the cause of her 6th nerve palsy was confirmed by the neuro-opthalmologist. Her symptoms resolved with the control of blood glucose, use of glucocorticoids and topical clonidine which has been proved useful in the treatment of diabetic gastroparesis and enteropathy.[br]Our patient is unique in that she has had 3 distinct simultaneous occurences of diabetic gastroparesis and mononeuritis with 6th nerve palsy during the course of her illness. Good control of blood glucose and use of steroids and topical clonidine is useful in the management of such attacks.[br][br]Nothing to Disclose: SS, GB, HW, DG, AS, PT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 16 299 2104 MON-183 PO47-01 Monday 1772 2012


1769 ENDO12L_MON-184 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Postural Orthostatic Tachycardia Syndrome Treatment with Liraglutide Aashish Samat, Robert Zimmerman Cleveland Clinic, Cleveland, OH [bold]Introduction:[/bold] Several medications from Beta blockers to octreotide have been used for the treatment for Postural orthostatic tachycardia syndrome (POTS). We describe a patient with resistant POTS who had resolution of symptoms after treatment with Liraglutide.[br][bold]Case:[/bold] A 19 year old female with panhypopituitarism and Diabetes Insipidus post surgery for a craniopharyngioma at age 12 on appropriate replacement developed symptoms of dizziness, on a daily basis, on upright posture which improved in a seated position a year post operatively. These spells occurred up to 8 times per day lasting 2-15 minutes. On exam, BP 120/78 HR 98 supine and BP 124/88 HR 120 standing. ENT, cardiac exam,coordination, tandem gait and Romberg[apos]s tests were normal. On echocardiogram her ejection fraction (EF) was 60 %. Tilt table test results: BP 120/72 HR 101 supine. On 70 degree tilt she developed dizziness with BP of 118/66 and HR 123. EKG was normal. On radionuclide hemodynamic testing :stroke volume 67 mls, HR 98 with a cardiac index of 3.8 L/m/min2 when supine. On standing cardiopulmonary to peripheral blood volume increased by 3 %, suggesting adequate veno constriction. HR increased to 119 and cardiac index decreased to 2.8 l/min/min 2. Her total blood volume was normal. She was diagnosed with POTS and was treated with metoprolol 50 mg bid without symptomatic improvement. Seven years later she presented with weight gain without evidence of diabetes and was started on Liraglutide 1.2 mcg/day for weight loss. On her following visit her symptoms of dizziness had abated and no postural tachycardia on exam. She lost 4 lbs (2.2% of total body weight) and had no symptoms of nausea and vomiting.[br][bold]Discussion[/bold][bold]:[/bold] There are GLP [ndash] 1 receptors present in the gastrointestinal tract, heart and vascular smooth muscle [sup]1[/sup].This suggests that GLP-1 agonist may have effects on hemodynamic parameters. In post myocardial infarction patients who received GLP-1 agonist infusion had improved EF [sup]2[/sup]. In patients who were undergoing CABG (Diabetic and non [ndash]diabetic) GLP-1 infusion reduced the use of inotropic and vasoactive infusion [sup]3-4[/sup]. The reduction in our patient[apos]s pulse could be secondary to improvement in her inotropic function along with the minimal weight loss. The cardiac effects of GLP-1 agonist are complex and need further long term studies to see if the effect of the change in pulse is related to weight loss or an independent phenomenon.[br][br]1 Ban K, et al. Circulation 2008; 117:2340-2350. 2 Nikolaidis LA, et al. Circulation 2004; 109:962-965. 3. Sokos GG, et al. Am J Cardiol 2007; 100:824-829. 4. Mussig K, et al. Am J Cardiol 2008; 102: 646-647.[br][br]Disclosures: RZ: Speaker, Novo Nordisk. Nothing to Disclose: AS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1353 299 2105 MON-184 PO47-01 Monday 1773 2012


1770 ENDO12L_MON-185 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Prolonged Deterioration of Glycemic Control and Marked Insulin Resistance after Only Two Doses of Betamethasone during Pregnancy Swapna A Dharashivkar, Osman M Shariff, Sri Prakash L Mokshagundam University of Louisville, Louisville, KY; University of Louisville, Louisville, KY Background: Betamethasone is widely used in pregnancy to enhance fetal lung maturity. We report an unusual case of persistent high glucose and marked IR after only 2 doses of betamethasone in the third trimester. Previous reports have only documented transient increase in glucose and insulin resistance.[br]Case: A 23 year old Hispanic lady with Class B diabetes mellitus was admitted with 32 weeks singleton gestation for oligohydramnios. Patient had been managed with oral glyburide 5 mg twice daily with optimal glucose control. In the third trimester, patient[apos]s HbA1c was 7.2% and she was switched to NPH insulin 28U twice daily and glulisine 16U with breakfast and 12U with lunch with blood glucose at goal. At 35 weeks she received 2 doses of betamethasone IM in anticipation of preterm delivery. Following this, patient had marked deterioration in glycemic control which persisted through pregnancy. Insulin was titrated and she was continued through pregnancy on U-500 R 130U three times a day and NPH insulin 35U at bedtime with excellent control. Patient had spontaneous vaginal delivery at 37 weeks and 2 days. Post-delivery, she did not require any insulin and went home on glyburide 5mg twice daily.[br]Major physiologic and metabolic adaptations during pregnancy maintain normal glucose homeostasis in mother. During the third trimester, IR results from several factors including changes in nutrient intake, altered adipokine/cytokine secretion (high leptin and TNFa, low adiponectin), high free fatty acids, and increased cortisol secretion. Glucose homeostasis is maintained by an increase in insulin secretion mediated by placental lactogens/somatogens. Studies in islets have shown that placental lactogens/somatogens and glucocorticoids have opposing effects on islet gene expression. Prolactin decreased Fork-head box protein o1, peroxisome proliferator activator receptor (PPAR)-gamma and carnitine-palmitoyl-transferase 1 (CPT-1) mRNAs, whereas dexamethasone increased FoxO1, PPAR-gamma coactivator-1a, and uncoupling protein 2 mRNAs. They differentially regulate beta-cell Glut-2 expression, fatty acid oxidation and glucose stimulated insulin secretion. We postulate that in our patient the administration of 2 doses of betamethasone compromised the delicate balance between these opposing effects which reverted only after pregnancy. The possibility of persistent and severe insulin resistance even with a short course of steroids and the role of U-500 R need to be kept in mind.[br][br]Nothing to Disclose: SAD, OMS, SLM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1122 299 2106 MON-185 PO47-01 Monday 1774 2012


1771 ENDO12L_MON-186 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Severe Hyperinsulinemic Hypoglycemia in an End-Stage Renal Patient on Peritoneal Dialysis and Normalization of Glycemia with Diazoxide Treatment: A Case Report and Literature Review Diep Dinh Nguyen, John T O[apos]Brian, Romesh Khardori Eastern Virginia Medical School, Norfolk, VA [bold][italic]Objective: [/italic][/bold]To report a case of severe hyperinsulinemic hypoglycemia in an ESRD patient on peritoneal dialysis with improvement of hypoglycemia following introduction of Diazoxide.[br][bold][italic]Clinical Case: [/italic][/bold][br]60 year-old male had bipolar disease and subsequent lithium toxicity, which resulted in ESRD, requiring hemodialysis starting in 2000, followed by peritoneal dialysis for the past 18 months. He initially presented to the ER for hypoglycemia with blood glucose in the 30[apos]s and lethargy. Peritonitis was ruled out, and the patient was discharged, only to be admitted a few days later with unconscious hypoglycemia as verified by the EMS. His recurrent episodes of symptomatic hypoglycemia always occurred after the completion of peritoneal dialysis with 2.5% dextrose dialysate (no icodextrin dialysate). Despite constant administration of D10W, patient[apos]s blood sugar continued to fall, and he required frequent administration of D50. His baseline adrenal function, thyroid function, growth hormone, IgF-1, and serum ammonia, were normal. At a time when the patient[apos]s blood sugar was 29mg/dL, a simultaneous C peptide level was 14ng/mL, total insulin was 21.4mcU/mL, free insulin was 18.5mcU/mL, and pro-insulin was 30.5pmol/L. Similar results were obtained during other hypoglycemic episodes. CT of abdomen/pelvis, Octreotide scan, and endoscopic ultrasound all were done without tumor localization. Given the severity of hypoglycemia, and biochemical markers that strongly suggested endogenous hyperinsulinemia, we started the patient on Diazoxide. Within 48 hours, the patient no longer required D50 or D10 IVF. His mental status returned to baseline with the normalization of his blood sugars.[br][bold][italic]Discussion: [/italic][/bold]The diagnosis of insulinoma is extremely difficult in the presence of concomitant renal failure. There are only 3 cases in the English literature where a tumor has been identified in this setting, one of whom was a patient on peritoneal dialysis whose symptoms were unmasked by dialysis suspension prior to surgery. A single case report was found of hyperinsulinemic hypoglycemia in ESRD in whom a tumor could not be located but was successfully treated with Diazoxide. That case occurred in a patient on hemodialysis, and our case extends this approach to patient on peritoneal dialysis.[br][bold][italic]Conclusion: [/italic][/bold]This is the first case report of the use of Diazoxide as treatment for hyperinsulinemia and recurrent hypoglycemia in an ESRD patient on peritoneal dialysis.[br][br](1) Service FJ. Nat Clin Pract Endocrinol Metab. 2006 Aug; 2(8):467-70. (2) Shaer AJ. Nephron. 2001 Nov; 89(3):337-9. (3) Basu A et al., J Clin Endocrinol Metab. 2002 Nov; 87(11):4889-91. (4) Matas AJ et al., Lancet. 1975 Apr 19;1(7912):883-6. (5) Jarrett RA et al., South Med J. 2009 Feb; 102(2):214-5. (6) Wang AY. Semin Nephrol. 2011 Mar; 31(2):159-71.[br][br]Nothing to Disclose: DDN, JTO, RK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 722 299 2107 MON-186 PO47-01 Monday 1775 2012


1772 ENDO12L_MON-187 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) To Eat or Not To Eat? Approach to Severe Liver Failure in a Patient with Anorexia Nervosa Markus Angelo Agito, Rommel Geronimo, Daniela Ciltea Akron General Medical Center, Akron, OH Introduction: Several studies have described various physical complications of anorexia nervosa (AN) but reports of severe liver damage resulting from AN are uncommon but have been reported. Severely malnourished patients presenting with liver failure have been associated with poor prognosis (1).[br]Case Report: A 29 year old female with a known history of AN was admitted after being found unconscious and severely hypoglycemic (glucose [lt]30mg/dL) at home. She was hypotensive, bradycardic and cool to touch. Mental status improved within minutes after administration of D5W. The patient appeared severely emaciated and dehydrated.[br]Blood glucose was normal (90mg/dL) upon arrival to the emergency department. Markedly elevated liver aminotransferase levels (AST 3260U/L, ALT 1825U/L, ALP 284U/L) were also noted. She had a normal hepatic panel from her previous admission. The liver was of uniform echogenicity with no focal hepatic lesions noted. Drug induced, viral, and autoimmune hepatitis were ruled out. The patient was initiated on D10W. A diet of 700kcal/day was also started and was slowly increased daily to prevent refeeding syndrome. Phosphorus was closely monitored and replaced as needed. Aminotransaminases improved gradually. When the supplemental diet was started on the 10[sup]th[/sup] hospital day, marked elevation of ALT was noted again while AST had only minimal elevation. ALT seemed to improve after stopping oral ingestion for a few days, then oral intake was gradually increased as tolerated until aminotransferases returned to normal.[br]Discussion: The markedly elevated aminotransaminase observed in this patient was most likely due to hypoperfusion of the liver and severe malnutrition. Severe calorie deficiency with continuous restriction of calorie intake deprived the patient[apos]s body of essential substrates required for the liver to perform its task in metabolism. The body resorts to protein and fat catabolism from organs other than the liver, such as the brain, heart, gut, and muscles resulting in atrophy. Depletion of hepatic glycogen, defective gluconeogenesis and failure of glucagon secretion subsequently results in hypoglycemia (2).[br]Conclusion: Proper nutritional support and respective timing should be carefully considered in severely malnourished individuals with markedly elevated aminotransaminases. Abnormal body composition can only be corrected safely when the cellular machinery has been adequately repaired.[br][br](1) Ozawa Y, Shimizu T, Shishiba Y. Elevation of serum aminotransferase as a sign of multiorgan-disorders in severely emaciated anorexia nervosa. Intern Med 1998;37:32[ndash]9. (2) Mehler PS, Krantz M. Anorexia nervosa medical issues. J Womens Health (Larchmt) 2003; 12:331.[br][br]Nothing to Disclose: MAA, RG, DC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2116 299 2108 MON-187 PO47-01 Monday 1776 2012


1773 ENDO12L_MON-188 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Case Study of Two Phenylketonuria Sisters: Too Late To Be Diagnosed Xiaomei Liu, Bingyin Shi, Hui Guo, Mahesh Dahal First Affiliated Hospital of Xi[apos]an Jiaotong University School of Medicine, Xi[apos]an, China The significant mental retardation was first noticed when both sisters were in their three years of age. However, no biochemical disorders were found through a routine biochemical tests, including liver and kidney functions, lipoprotein, urine and blood cell count analysis. Progressively, both sisters were behaving in a strange way, with change in personality and altered sleep cycle. During last seven years, their poor condition kept on and failed to respond to the medical advice from numerous doctors. The younger sister had take the Tab. Risperidone for one year due to mental disorder. After then both sisters were diagnosed as hereditary leukodystrophy by the magnetic resonance imaging and neurophic agent was tried but in vain. Then, they were referred to our hospital seeking further expertise. After systematic examinations including FeCl3 test of urine, MILS Tandem Mass Spectrometry, MILS international test, characteristic pattern of urinary pterins and sleeping electroencephalogram, it was confirmed that the two sisters were suffered from phenylketonuria. The symptoms were alleviated after dietary restriction of phenylalanine and symptomatic treatment. The lesson learned from these two cases is that phenylketonuria must cross the brain when patient presents with mental retardation in their youth, and some screening tests need to be done promptly.[br][br]Nothing to Disclose: XL, BS, HG, MD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 345 299 2109 MON-188 PO47-01 Monday 1777 2012


1774 ENDO12L_MON-189 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) An Adult Case of Fructose-1,6-Bisphosphatase (FBPase) Deficiency with Novel Compound Heterozygous Mutations in FBP1 Gene Ikki Sakuma, Hidekazu Nagano, Seiichiro Higuchi, Tomoko Takiguchi, Sawako Suzuki, Tomohiko Yoshida, Koutaro Yokote, Tomoaki Tanaka Chiba University Hospital, Chiba, Japan Fructose-1,6-bisphosphatase (FBPase, EC3.1.3.11) deficiency is an autosomal recessive disorder associated with hypoglycemia and metabolic acidosis caused by a mutation of FBP gene (FBP1). FBPase is a key regulatory enzyme in gluconeogenesis, which catalyzes the hydrolysis of fructose 1,6-bisphosphate to fructose 6-phosphate and inorganic phosphate. Therefore, fasting and febrile infections are known to trigger life-threatening episodes of hypoglycemia and lactic acidosis during newborn infants and infancy, while these episoidic spells are rarely seen in adulthood. Here we describe a 22-year-old female of FBPase deficiency who presented severe hypoglycemic attack with lactic acidosis in emergency room and then was admitted to our hospital. She had similar episodes of severe hypoglycemia and acidosis triggered by febrile illness until early childhood. Blood examination including BGA on admission displayed the very low level of blood glucose (0.67 mM) and severe lactic acidosis (pH 6.845, base excess -28.2 mmol/L, HCO3- 2.5 mmol/L and lactate 13.1 mmol/L). She was treated with intravenous administration of glucose and sodium bicarbonate as well as continuous hemodiafiltration and then recovered very well within a couple of days. To clarify and make a diagnosis, we examined the profile of blood acylcarnitine and urinary organic acids. While there was no specific pattern in acylcarnitine profile implicating in a normal beta-oxidation pathway, we found significant elevation of lactate, pyruvate, 3-hydroxybutyrate and glycerol, indicating FBPase deficiency. Consistently, oral fructose tolerance test demonstrated rapid decrease of blood glucose with increase of lactate concentrations. Further, genetic analyses revealed that the patient carried compound heterozygote missense mutations of FBP1 with G164D and F194S, a novel and previously reported site, respectively. Thus, we report an adult case of FBPase deficiency with novel compound heterozygous mutations.[br][br]Nothing to Disclose: IS, HN, SH, TT, SS, TY, KY, TT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 481 299 2110 MON-189 PO47-01 Monday 1778 2012


1775 ENDO12L_MON-190 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Low-Dose (15 mg/day) Pioglitazone Improves Glycemic Control and Insulin Secretion/Insulin Sensitivity Index with Minimal Weight Gain in Type 2 Diabetes Zandra Perez-Cadena, Giuseppe Daniele, Alberto Chavez, Lauren Cortez, Amalia Gastaldelli, Ralph DeFronzo, Franco Folli, Devjit Tripathy University of Texas Health Science Center at San Antonio, San Antonio, TX; Audie L Murphy Veterans Hospital, San Antonio, TX Pioglitazone (PIO) is a potent insulin-sensitizer but its use is limited by weight gain and fluid retention at higher doses (30-45mg/day). The aim of the study was to evaluate whether low dose PIO (15mg/day) could minimize the weight gain while retaining the metabolic effects. We examined the effects of low-dose PIO (15mg/day) on metabolic parameters and body composition in 20 T2DM subjects (age 56 [plusmn]1 yr, BMI 33.5 [plusmn]1.3 kg/m[sup]2[/sup], FPG =145 [plusmn]8.3mg/dl, HbA[sub]1c[/sub] 7.5%) randomized to PIO (n=11, M/F 7/4, 15mg/day) or PLAC (n=9, M/F 7/2) who participated in an OGTT with insulin and C-peptide measured every 30 min, euglycemic-hyperinsulinemic clamp (80mU/m[sup]2[/sup]-min) with measurement of body fat (BF) by DXA at 0 and 24 wks. Insulin secretion was measured by deconvolution analysis from C-peptide (insulin secretory rate) during OGTT. At baseline, subjects in both groups had similar age, BMI, % BF, HbA[sub]1c[/sub], and fasting and 2-h plasma glucose. PIO led to a greater reduction in fasting PG (140 [plusmn]11 to 118 [plusmn]9.5 vs 150 [plusmn]13 to 140 [plusmn]12 mg/dL, p=0.004), 2-h PG (265 [plusmn]16 to 243 [plusmn]15 vs 285 [plusmn]12 to 293 [plusmn]10 mg/dL, p=0.02), HbA[sub]1c[/sub] (7.0 [plusmn] 0.2 to 6.5 [plusmn] 0.2 vs 8.0 [plusmn] 0.5 to 7.7 [plusmn] 0.5%, p=0.02), and triglyceride levels (190 [plusmn]16 to 135 [plusmn]17 vs 207 [plusmn]49 to 180 [plusmn]44 mg/dL, (p=0.005). PIO improved whole-body insulin sensitivity (IS) (3.2 [plusmn] 0.5 to 4.1 [plusmn] 0.5 vs 3.4 [plusmn]0.5 to 4.5 [plusmn]0.5 mg/kg/min, p=0.005). Improved IS was primarily due to improvement in non-oxidative glucose metabolism (1.6 [plusmn]0.5 to 3.7 [plusmn]0.5 mg/kg/min, p=0.05). The disposition index (insulin secretion/insulin sensitivity index) (ISR x M-value) improved with PIO (13.8 [plusmn]4.2 to17.0 [plusmn]3.9, p=0.05) but not with PLAC. Body weight and % BF increased by 1.8 [plusmn]0.5kg and 2%, respectively (p[lt]0.05), vs PLAC in patients treated with PIO, which was considerably lower than that reported with higher doses of PIO. Despite similar insulin levels, a greater suppression of plasma FFA concentrations was noted during insulin clamp following therapy with PIO (0.066 [plusmn]0.008 vs 0.082 [plusmn]0.007mM, p=0.04) suggesting greater improvement in adipocyte insulin resistance. In conclusion, low-dose PIO therapy was well tolerated and improved glycemic control, lipid profile and whole-body and adipose insulin resistance with minimal weight gain. Low dose PIO may be considered for long[ndash]term treatment for prevention of T2DM or as an add-on therapy for T2DM.[br][br]Sources of Research Support: Takeda Pharmaceutical Company; Bartter Research Unit at Audie L. Murphy Veterans Hospital.[br][br]Disclosures: RD: Research Funding, Takeda. FF: Research Funding, Takeda. Nothing to Disclose: ZP-C, GD, AC, LC, AG, DT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2095 300 2111 MON-190 PO11-01 Monday 1779 2012


3038 ENDO12L_S43-3 SYMPOSIUM SESSION: TRANSLATIONAL - Adipose Tissue Inflammation [amp] Remodeling (3:45 PM - 5:15 PM) Adipose Tissue Fibrosis in Human Karine Clement Cordelier Research Center, Paris, France Extracellular matrix remodeling is essential for tissue growth and expansion. However overproduction of collagens and other extracellular matrix components causes a common condition known as fibrosis which alters tissue structure and functions. Recent observations have shown that obesity is associated with a pathological accumulation of extracellular matrix proteins leading to fibrosis in the adipose tissue. Our team found major changes in the expression of genes encoding extracellular matrix components in human obese adipose tissue and in response to weight loss (1). Combined histopathological evaluation further revealed increased amounts of fibrosis in obese vs. lean adipose tissue. This was found both in subcutaneous and visceral adipose depots (2). More specifically, obesity associates with marked fibrotic deposition around adipocytes. Pericellular fibrosis associates negatively with adipocyte size (3). Increased fibrosis is also linked with lower fat mass loss after gastric surgery. However the local and systemic consequences of adipose tissue fibrosis are not fully understood. In several human studies, high collagen VI expression was also associated with increased macrophage accumulation in adipose tissue. Adipose tissue fibrosis might result, at least in part, from chronic low-grade inflammation that characterizes adipose tissue in obesity while the kinetic of events need to be precisely defined. On a fundamental basis, adipose tissue fibrosis could provoke a mechanical stress which deeply alters adipocyte biology. Fibrosis now represents a relatively new determinant of the biological alteration observed in adipose tissue of obese subjects Thus, the identification of cellular and molecular actors relaying the development of fibrosis could provide new targets to improve adipose tissue function in obesity. This lecture will provide new insights in this rapidly evolving field which is the pathological alteration of adipose tissue in obesity with a specific focus on adipose tissue fibrosis in human studies.[br][br](1) Henegar C, et al. Adipose tissue transcriptomic signature highlights the pathological relevance of extracellular matrix in human obesity. Genome Biol 2008 9:R14. (2) Divoux A, et al Fibrosis in human adipose tissue: composition, distribution, and link with lipid metabolism and fat mass loss. Diabetes 2010 59:2817-25. (3) Tam CS et al. Adipose tissue remodeling in children: the link between collagen deposition and age-related adipocyte growth. JCEM, In press.[br][br]Sources of Research Support: European Grant 7th Framework ADAPT, National Agency of Research ADIPOFIB.[br][br]Nothing to Disclose: KC 2012-06-25T16:45:00 Room 362 2012-06-25T00:00:00 1899-12-30T16:45:00 39 335 2634 S43-3 T04-S04 Monday 329 2012


3039 ENDO12L_S44-1 SYMPOSIUM SESSION: TRANSLATIONAL - Bone Diseases in Cancer Patients (3:45 PM - 5:15 PM) How Is Bone Compromised in Cancer Patients? Jean-Jacques Body CHU Brugmann, Brussels, Belgium Patients with endocrine-related cancers are notably at increased risk for developing osteoporosis as a complication from their adjuvant treatment, especially aromatase inhibitors (AIs) in early breast cancer (EBC) and androgen deprivation therapy (ADT) in early prostate cancer (EPC). Women with BC, even in the absence of skeletal metastases, are known to have a higher incidence of fractures than women of the same age without BC. In a prospective analysis of postmenopausal women from the Women[apos]s Health Initiative Observational Study (WHI-OS), fracture rates for BC survivors were increased by 68.6 fractures per 10,000 person-years compared with women without BC. The increased risk of fracture was significant for women with a BC diagnosis regardless of age (HR [sim]1.3; P [lt] .001), and was not limited to asymptomatic vertebral fractures. Chemotherapy is associated with increased fracture risk in premenopausal women with EBC because it can induce early menopause and may have direct, toxic effects on bone cells. The increased risk of fracture in EBC patients has become even more evident following the increased use of AIs as adjuvant therapy. AI-induced bone loss (AIIBL) occurs at a rate at least 2-fold higher than bone loss seen in healthy, age-matched postmenopausal women and they have a more than 30% higher risk of fractures. Recently considered fracture risk factors in EBC are AI therapy, T-score [lt][ndash]1.5, age [gt]65 yrs, family history of hip fracture, history of fragility fracture after age 50, oral corticosteroid use [gt]6mo, smoking, and BMI [lt]20 kg/m2. The WHO-FRAX algorithm does not address AIIBL and may underestimate EBC fracture risk. Bisphosphonates and denosumab can preserve BMD during AI therapy for EBC. Poor compliance may reduce oral bisphosphonate benefits. Overall, bisphosphonate evidence is strongest for zoledronic acid (ZOL; 4mg q6mo). Potential anticancer activity of ZOL might provide benefits beyond preserving bone mass. Patients initiating AIs with T-score [lt][ndash]2.0 or [ge]2 risk factors should receive antiresorptive therapy. In prostate cancer, bone loss that occurs with ADT is also more rapid and severe than that associated with normal age-related bone loss. ADT increases fracture risk and the hazard ratios appear to be comparable to the ones reported for AIs in breast cancer. Bisphosphonates, notably ZOL, can prevent bone loss under ADT. Denosumab (60 mg sc twice a year) prevents ADT-induced bone loss but also decreases incidental vertebral fractures.[br][br]Nothing to Disclose: J-JB 2012-06-25T15:45:00 Room 320 2012-06-25T00:00:00 1899-12-30T15:45:00 2424 336 2635 S44-1 T03-S03 Monday 331 2012


3040 ENDO12L_S44-2 SYMPOSIUM SESSION: TRANSLATIONAL - Bone Diseases in Cancer Patients (3:45 PM - 5:15 PM) Mechanisms of Action of Bone Preserving Drugs in Cancer Mone Zaidi Mt Sinai School of Medicine, New York, NY Disclosure Incomplete: MZ 2012-06-25T16:15:00 Room 320 2012-06-25T00:00:00 1899-12-30T16:15:00 6035 336 2636 S44-2 T03-S03 Monday 332 2012


3041 ENDO12L_S44-3 SYMPOSIUM SESSION: TRANSLATIONAL - Bone Diseases in Cancer Patients (3:45 PM - 5:15 PM) Optimizing Bone Health in Cancer Patients Robert E Coleman CR-UK/YCR Sheffield Cancer Research Centre, Sheffield, UK [bold]Preserving bone health:[/bold] There are increasing numbers of long-term survivors from cancer, and many of these individuals are at increased risk of osteoporosis, largely due to the endocrine changes induced by treatment characterised by a reduction in the level of bio-available oestradiol. The use of bone-targeted treatments in early cancer has become increasingly important to prevent these adverse effects of cancer treatments on bone health. These include chemotherapy induced ovarian failure across a range of curable malignancies, ovarian suppression and aromatase inhibitors in breast cancer patients, and long-term androgen deprivation therapy (ADT) in men with prostate cancer. Bisphosphonate strategies, similar to those used to treat post-menopausal osteoporosis, are the intervention of choice for patients with low bone mineral density or rapid bone loss, along with adequate calcium and vitamin D intake and a healthy lifestyle. The results of several intervention studies, with either bisphosphonates or denosumab in early breast cancer and following ADT in prostate cancer, indicate that bone mineral density is maintained and bone turnover rates normalized.[br][bold]Preventing metastasis:[/bold] Interactions between cancer cells in the bone marrow microenvironment and normal bone cells are of fundamental importance in cancer metastasis. Factors secreted by tumour cells stimulate stromal cells and osteoblasts to release receptor activator of NF-kB ligand (RANKL), which binds to RANK on the surface of precursor and mature osteoclasts, leading to stimulation of osteoclastic bone resorption. This in turn results in release of bone derived growth factors that may provide a more favourable environment for cancer cells to survive and proliferate. Bone targeted treatments may modify the course of the disease and disrupt the metastatic process via inhibitory effects on this [ldquo]vicious cycle[rdquo]. In the adjuvant setting, there is increasing clinical trial support for this therapeutic strategy, especially in the context of low levels of circulating reproductive hormones. Improvements in disease free and overall survival have been reported recently in several clinical trials of adjuvant bisphosphonates in breast cancer and with denosumab in men with castrate resistant prostate cancer.[br][bold]Conclusions:[/bold] Bone targeted treatment is an important component of cancer care to reduce cancer treatment induced bone loss, and may modify the course of the underlying malignancy is some clinical settings.[br][br]Disclosures: REC: Speaker, Amgen, Novartis Pharmaceuticals. 2012-06-25T16:45:00 Room 320 2012-06-25T00:00:00 1899-12-30T16:45:00 2416 336 2637 S44-3 T03-S03 Monday 333 2012


3042 ENDO12L_S45-1 SYMPOSIUM SESSION: CLINICAL - Controversial Issues in Cushing[apos]s Syndrome [quot]To Screen or Not To Screen? That Is the Question (3:45 PM - 5:15 PM) When Should We Screen? Antoine Tabarin Universite de Bordeaux II, Pessac, France Cushing[apos]s syndrome (CS) includes numerous clinical features reflecting the widespread distribution of glucocorticoid receptors in multiple target organs. Although CS is easily recognizable clinically when overt, one obstacle for its diagnosis is its broad spectrum of presentation. Symptoms like obesity, diabetes and hypertension are highly prevalent in CS but are also extremely common in the general population. Conversely, specific symptoms due to protein-wasting have a strong diagnostic power but may be subtle or lack in a number of patients. This spectrum of clinical presentation may lead to underdiagnosis and the true prevalence of CS may be higher than expected. A systematic search for occult CS has been performed in screening studies involving patients harboring common clinical symptoms belonging to CS but in whom no specific clinical feature was found. Seven prospective studies conducted in diabetic patients have revealed a prevalence of occult CS ranging from 0.0 to 9.4% with a pooled estimate around 2%. One prospective study evaluated patients with osteoporosis identified occult CS in 4.8 to 10.8% of patients. Screening studies in obese patients have provided controversial results. Retrospective studies, involving limited number of patients found occult CS in 3.5% to 8.7% of cases while a large scale cross-sectional prospective US study did not identify a single case of CS. Two methodologically imperfect studies conducted on large cohorts of hypertensive patients identified 0.5% to 2% of CS. Thus, systematic screening studies performed mainly in obese type 2 diabetic patients have revealed an unexpected prevalence of occult CS that is clearly greater than that quoted in population-based studies. These findings may provide a rationale for a systematic screening for CS in this patient population. However, a screening strategy is only justified if it is supported by evidence of its efficacy and if benefits will outweigh drawbacks. The burden of the targeted condition (frequency and severity) should justify action. Natural history should include a latent or preclinical stage to allow early diagnosis and intervention. The accuracy of diagnostic tests must be high. Detection of the disease is only useful if treatment improves clinical outcomes. Finally, the screening program must be acceptable by professionals and the health-care system, in terms of work load and costs. These points will be discussed according to data of the literature.[br][br]Nothing to Disclose: AT 2012-06-25T15:45:00 Theater A 2012-06-25T00:00:00 1899-12-30T15:45:00 2536 337 2638 S45-1 T01-S04 Monday 335 2012


3012 ENDO12L_S42-3 SYMPOSIUM SESSION: CLINICAL - State of the Art Management of Nodular Thyroid Disease (9:30 AM - 11:00 AM) The Natural History of the Benign Nodule Sebastiano Filetti University of Rome La Sapienza, Rome, Italy The past 2 decades have witnessed a steady rise in the frequency of nodular thyroid disease, which is an emerging public health concern. This trend stems largely from improved detection of tiny nodules thanks to enhanced diagnostic activity in clinical practice. The discovery of any thyroid lesion is naturally stressful for the patient, but most of the nodules being discovered are clinically insignificant. Very few require surgical treatment (for suspected or proven malignancy or relief of neck discomfort). But once these conditions have been excluded, physicians are faced with a dismaying variety of nonsurgical management options, including simple surveillance. The lack of consensus regarding optimal management of these nodules reflects our incomplete knowledge of their natural history. Treatment advocates argue that untreated nodules will grow, but in a retrospective study of 140 untreated patients followed 15 years, only 14% of the benign nodules displayed any growth at all (1). In another study, 139 cytologically benign nodules were followed without treatment for 3 years (mean), and over half displayed sonographically-documented volume increases of [underline][gt][/underline]30% (2). Similarly, increases of [underline][gt][/underline]15% were reported in 39% of the benign nodules followed for 20 months (mean) (3). Collectively, these data point to the existence of a subgroup of benign nodules that can be expected to grow over time, in general very slowly. The challenge is to find markers that can predict this type of behavior. The factors influencing thyroid nodule growth are currently unknown, and their identification will require large prospective studies. In Italy, a multicenter observational study of this type has been underway since 2006. Its aim is to characterize the natural history of 1569 untreated, cytologically benign thyroid nodules (993 patients). In median follow-up of 4.5 years (range: 4-5), [lt] 15% of these nodules have increased in size, and higher growth rates appear to be associated with multinodular goiter, maximum nodule diameters [gt] 1 cm, and diagnosis before age 45. These findings could prove useful for identifying thyroid nodules that need closer surveillance.[br][br](1) Kuma K et al., World J Surg 1992; 16:583. (2) Quadbeck B et al., Exp Clin Endocrinol Diabetes 2002; 110:348. (3) Alexander EK et al. Ann Intern Med 2003; 138:315.[br][br]Nothing to Disclose: SF 2012-06-25T10:30:00 Theater A 2012-06-25T00:00:00 1899-12-30T10:30:00 2501 266 1868 S42-3 T02-S03 Monday 298 2012


3014 ENDO12L_CP2 CLINICAL PRACTICE GUIDELINES: CLINICAL - Management of Hyperglycemia in Hospitalized Patients in Non-Critical Care Settings (11:15 AM - 12:00 PM) Moderator Susan Braithwaite TBD, Evanston, IL Session supported by: Medtronic Diabetes, Merck [amp] Co., Inc. and sanofi-aventis U.S.[br][br]Disclosure Incomplete: Author to be determined 2012-06-25T11:15:00 Grand Ballroom AB 2012-06-25T00:00:00 1899-12-30T11:15:00 6268 278 1935 CP2 CPG2 Monday 300 2012


3015 ENDO12L_CDW5-1 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Negotiations: Basic [amp] Clinical Research (11:15 AM - 12:15 PM) Negotiations: Basic [amp] Clinical Research Deborah Jean Good Virginia Tech, Blacksburg, VA Many job seekers accept the first offer they receive, and later regret not asking for specific things, such as increased pay, technician assistance, needed equipment and even moving costs, all of which could have helped them to better succeed in their new position. Deborah J. Good, an Associate Professor at Virginia Tech, has served on numerous job search committees, and has negotiated several different jobs and job offers[ndash]some better than others! This workshop will use the speaker[apos]s personal experience to examine some of the dos and don[apos]ts of job offer negotiations. Job seekers at the assistant and associate professor level, as well as graduate students looking towards their first postdoctoral fellowship, are welcome to attend and bring their questions to this interactive workshop.[br][br]Session supported by: Novo Nordisk Inc. [br][br]Nothing to Disclose: DJG 2012-06-25T11:15:00 Trainee Career Center 2012-06-25T00:00:00 1899-12-30T11:15:00 2392 279 1936 CDW5-1 CDW3 Monday 301 2012


3016 ENDO12L_CDW5-2 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Negotiations: Basic [amp] Clinical Research (11:15 AM - 12:15 PM) Negotiations: Basic [amp] Clinical Research Fredric B Kraemer Stanford University School of Medicine, Stanford, CA Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: FBK 2012-06-25T11:15:00 Trainee Career Center 2012-06-25T00:00:00 1899-12-30T11:15:00 6251 279 1937 CDW5-2 CDW3 Monday 302 2012


3017 ENDO12L_MC3-1 MASTER CLINICIAN: CLINICAL - Difficult Pituitary Cases (12:15 PM - 1:15 PM) Difficult Pituitary Cases Mary Lee Vance University of Virginia Medical Center, Charlottesville, VA Disclosure Incomplete: MLV 2012-06-25T12:15:00 Grand Ballroom AB 2012-06-25T00:00:00 1899-12-30T12:15:00 6113 280 1938 MC3-1 MC3 Monday 304 2012


3018 ENDO12L_MC3-2 MASTER CLINICIAN: CLINICAL - Difficult Pituitary Cases (12:15 PM - 1:15 PM) Difficult Pituitary Cases Peter James Trainer The Christie NHS Foundation Trust, Manchester, UK Disclosures: PJT: Clinical Researcher, Ipsen, Versartis. 2012-06-25T12:15:00 Grand Ballroom AB 2012-06-25T00:00:00 1899-12-30T12:15:00 6114 280 1939 MC3-2 MC3 Monday 305 2012


3019 ENDO12L_CDW6 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Negotiations: Clinical Practice (12:30 PM - 1:00 PM) Negotiations: Clinical Practice Jason Adam Wexler , Washington, DC Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: JAW 2012-06-25T12:30:00 Trainee Career Center 2012-06-25T00:00:00 1899-12-30T12:30:00 6221 281 1940 CDW6 CDW4 Monday 306 2012


3020 ENDO12L_Y2 ENDOCRINE YEAR IN BASIC/CLINICAL SCIENCE: BASIC - The Year in Sex Differences (1:00 PM - 1:45 PM) The Year in Sex Differences Deborah J Clegg University of Texas Southwestern Medical Center, Dallas, TX Nothing to Disclose: DJC 2012-06-25T13:00:00 Room 332 2012-06-25T00:00:00 1899-12-30T13:00:00 6116 290 1950 Y2 YI2 Monday 307 2012


3021 ENDO12L_CMF9-1 CASE MANAGEMENT FORUM: CLINICAL - Management of Apparent Secondary Hypogonadism Due to Obesity, Opiates [amp] Anabolic Steroids (2:45 PM - 3:30 PM) Management of Apparent Secondary Hypogonadism Due to Obesity, Opiates [amp] Anabolic Steroids Bradley David Anawalt University of Washington, Seattle, WA Disclosure Incomplete: BDA 2012-06-25T14:45:00 Theater A 2012-06-25T00:00:00 1899-12-30T14:45:00 6169 323 2616 CMF9-1 CMF-MR2 Monday 308 2012


3022 ENDO12L_CMF9-2 CASE MANAGEMENT FORUM: CLINICAL - Management of Apparent Secondary Hypogonadism Due to Obesity, Opiates [amp] Anabolic Steroids (2:45 PM - 3:30 PM) Management of Apparent Secondary Hypogonadism due to Obesity, Opiates [amp] Anabolic Steroids Richard Arthur Bebb University of British Columbia, Vancouver, Canada Nothing to Disclose: RAB 2012-06-25T14:45:00 Theater A 2012-06-25T00:00:00 1899-12-30T14:45:00 6182 323 2617 CMF9-2 CMF-MR2 Monday 309 2012


3240 ENDO12L1_7007 PRE-CONFERENCE EVENT: Introductory Hands-On Thyroid Ultrasound Workshop (1:30 PM - 6:30 PM) Introductory Hands-On Thyroid Ultrasound Workshop Registration and fee required 2012-06-22T13:30:00 George R. Brown Convention Center 2012-06-22T00:00:00 1899-12-30T13:30:00 7007 7 7 9905 Friday 0 2012


3241 ENDO12L1_7008 PRE-CONFERENCE EVENT: Trainee Reception (5:30 PM - 7:00 PM) Trainee Reception Open[br][br]Supported by Novo Nordisk, Inc. 2012-06-22T17:30:00 Hilton Ballroom of the Americas 2012-06-22T00:00:00 1899-12-30T17:30:00 7008 8 8 9906 Friday 0 2012


3242 ENDO12L1_7009 PRE-CONFERENCE EVENT: How To Secure Promotion and Tenior Reception (6:30 PM - 8:00 PM) How To Secure Promotion and Tenior Reception 2012-06-22T18:30:00 George R. Brown Convention Center 2012-06-22T00:00:00 1899-12-30T18:30:00 7009 10 10 9907 Friday 0 2012


3243 ENDO12L1_7010 PRE-CONFERENCE EVENT: Corporate Liaison Board Forum [ndash] Diabetes [amp] Cardiovascular Risk: Is Personalized Medicine the Answer? (6:30 PM - 9:30 PM) Corporate Liaison Board Forum [ndash] Diabetes [amp] Cardiovascular Risk: Is Personalized Medicine the Answer? The Endocrine Society[apos]s Corporate Liaison Board 2012-06-22T18:30:00 Hilton Lanier Ballroom 2012-06-22T00:00:00 1899-12-30T18:30:00 7010 9 9 9908 Friday 0 2012


3244 ENDO12L1_7011 IDEAL[reg] Live Learning Center (June 23-25 10:30 AM - 4:00 PM) IDEAL[reg] Live Learning Center Supported by Abbott Laboratories and Endo Pharmaceuticals Inc. 2012-06-23T10:30:00 George R. Brown Convention Center 2012-06-23T00:00:00 1899-12-30T10:30:00 7011 28 49 9909 Saturday 0 2012


3149 ENDO12L_M8A MEET-THE-PROFESSOR: CLINICAL - Dietary Issues in Ethnic Groups (5:30 PM - 6:15 PM) Dietary Issues in Ethnic Groups Abhimanyu Garg University Texas Southwest Medical Center, Dallas, TX Nothing to Disclose: AG 2012-06-23T17:30:00 Room 362 2012-06-23T00:00:00 1899-12-30T17:30:00 6181 107 909 M8 D9A Saturday 70 2012


3150 ENDO12L_M9A MEET-THE-PROFESSOR: CLINICAL - Difficult Thyroid Function Tests (5:30 PM - 6:15 PM) Difficult Thyroid Function Tests Douglas S Ross Massachusetts General Hospital, Waban, MA Disclosures: DSR: Speaker, Genzyme Corporation; Researcher, Genzyme Corporation. 2012-06-23T17:30:00 Room 332 2012-06-23T00:00:00 1899-12-30T17:30:00 6161 108 910 M9 TH4A Saturday 71 2012


3151 ENDO12L_M10A MEET-THE-PROFESSOR: CLINICAL - Hyponatremia (5:30 PM - 6:15 PM) Hyponatremia Joseph G Verbalis Georgetown University, Washington, DC Disclosures: JGV: Consultant, Astellas, Cardiokine, Otsuka, Sanofi-Aventis. 2012-06-23T17:30:00 Room 361 ABDE 2012-06-23T00:00:00 1899-12-30T17:30:00 6166 109 911 M10 PIT3A Saturday 72 2012


3152 ENDO12L_M11A MEET-THE-PROFESSOR: CLINICAL - Insulin Pumps (5:30 PM - 6:15 PM) Insulin Pumps Howard Wolpert Joslin Diabetes Center, Boston, MA Session supported by: Lilly USA, LLC and Medtronic Diabetes[br][br]Disclosure Incomplete: HW 2012-06-23T17:30:00 Room 342 ABDE 2012-06-23T00:00:00 1899-12-30T17:30:00 6249 110 912 M11 D4A Saturday 73 2012


3153 ENDO12L_M13A MEET-THE-PROFESSOR: CLINICAL - Non-Cancer Thyroid [apos]Hot Topics[apos] in Children [amp] Adolescents (5:30 PM - 6:15 PM) Non-Cancer Thyroid [apos]Hot Topics[apos] in Children [amp] Adolescents Stephen H LaFranchi Oregon Health Sciences University, Portland, OR Nothing to Disclose: SHL 2012-06-23T17:30:00 Room 310 2012-06-23T00:00:00 1899-12-30T17:30:00 6167 111 913 M13 PED5A Saturday 74 2012


3154 ENDO12L_M14A MEET-THE-PROFESSOR: CLINICAL - Non-PTH Hypercalcemia (5:30 PM - 6:15 PM) Non-PTH Hypercalcemia Gregory A Clines University of Alabama Birmingham, Birmingham, AL Nothing to Disclose: GAC 2012-06-23T17:30:00 Room 320 2012-06-23T00:00:00 1899-12-30T17:30:00 6141 112 914 M14 B2A Saturday 75 2012


3155 ENDO12L_M15A MEET-THE-PROFESSOR: CLINICAL - Premature Ovarian Failure ( Insufficiency) (5:30 PM - 6:15 PM) Premature Ovarian Insufficiency Nanette F Santoro University of Colorado School of Medicine, Aurora, CO Nothing to Disclose: NFS 2012-06-23T17:30:00 Room 352 DEF 2012-06-23T00:00:00 1899-12-30T17:30:00 6151 113 915 M15 FR5A Saturday 76 2012


3156 ENDO12L_CMF4-1A CASE MANAGEMENT FORUM: CLINICAL - Endocrine Disease [amp] Bone: Diagnostic [amp] Management Issues (1:00 PM - 1:45 PM) Endocrine Disease [amp] Bone: Diagnostic [amp] Management Issues Lynn Kohlmeier Spokane Osteoporosis, Spokane, WA Nothing to Disclose: LK 2012-06-24T13:00:00 Room 310 2012-06-24T00:00:00 1899-12-30T13:00:00 6143 161 1038 CMF4-1 CMF-B1A Sunday 119 2012


3157 ENDO12L_CMF4-2A CASE MANAGEMENT FORUM: CLINICAL - Endocrine Disease [amp] Bone: Diagnostic [amp] Management Issues (1:00 PM - 1:45 PM) Endocrine Disease [amp] Bone: Diagnostic [amp] Management Issues Jan M Bruder University of Texas HSC at San Antonio, San Antonio, TX Nothing to Disclose: JMB 2012-06-24T13:00:00 Room 310 2012-06-24T00:00:00 1899-12-30T13:00:00 6193 161 1039 CMF4-2 CMF-B1A Sunday 120 2012


3158 ENDO12L_CMF5-1A CASE MANAGEMENT FORUM: CLINICAL - Primary Hyperaldosteronism (1:00 PM - 1:45 PM) Primary Hyperaldosteronism Robert M Carey University of Virginia Health System, Charlottesville, VA Nothing to Disclose: RMC 2012-06-24T13:00:00 Room 362 2012-06-24T00:00:00 1899-12-30T13:00:00 6173 162 1040 CMF5-1 CMF-A2A Sunday 121 2012


2838 ENDO12L_S12-1 SYMPOSIUM SESSION: TRANSLATIONAL - BAT: Development [amp] Therapeutic Potential (3:45 PM - 5:15 PM) Control of Thermogenic BAT Function through Its Innervation Timothy J Bartness Georgia State University, Atlanta, GA Brown adipose tissue (BAT) is innervated by the sympathetic nervous system (SNS), the activation of which is the principal stimulator of BAT thermogenesis. We defined the central origins of the SNS outflow to BAT using the transneuronal retrograde viral tracer, pseudorabies virus (PRV) injected into interscapular BAT (IBAT). We combined PRV SNS outflow labeling with in situ hybridization for melanocortin receptor-4 receptor (MC4-R) mRNA demonstrating high colocalization ([sim]60%) across many neuroaxis sites, including the hypothalamic paraventricular nucleus (PVH) and the subZona incerta (subZI). MC3/4-R agonism by melanotan II (MTII) nanoinjection into the PVH, increased interscapular (IBAT) temperature (TIBAT) in freely moving Siberian hamsters bearing telemetric thermistors implanted under the IBAT., Nanoinjections of MTII or the specific MC4-R agonist (cyclo [[beta]-Ala-His-D-Phe-Arg-Trp-Glu]-NH2) into the subZI, an unappreciated node in the CNS SNS outflow to BAT, also significantly increased TIBAT. By contrast, subZI nanoinjection of the MC4-R antagonist, HS024 significantly decreased TIBAT alone and if pre-injected before the MC4-R agonist, blocked the agonist-induced increased TIBAT. BAT also has sensory innervation, as revealed by the transneuronal anterograde viral tract tracer, the H129 strain of herpes simplex 1 virus, with infected neurons appearing across the neuroaxis, especially the brainstem and forebrain, and occurring in many of the same sites participating in the SNS outflow to IBAT. Indeed, injection of both H129 to label central sensory circuits and PRV to label SNS outflow into IBAT resulted in doubly-infected neurons (SNS-sensory loops) in several brain areas including the PVH and raphe pallidus suggesting neural feedback loop regulation of the SNS drive to BAT. Local IBAT capsaicin injection to ablate small-unmyelinated sensory nerves impaired normal TIBAT indicating BAT sensory nerves help control its thermogenic function. Finally, crosstalk between BAT and white adipose tissue (WAT) was shown preliminarily by injecting H129 into WAT to label CNS sensory inputs to brain and PRV into IBAT to label brain SNS outputs suggesting lipid stores may affect BAT SNS drive. Shared CNS SNS outflow neurons to WAT and BAT also were seen preliminarily by injecting two PRV strains with unique reporters suggesting neurology underlying conditions when SNS drive increases to both tissues. Thus, complex neural control of BAT thermogenesis exists.[br][br]Sources of Research Support: NIH R01 DK078358.[br][br]Nothing to Disclose: TJB 2012-06-23T15:45:00 Theater C 2012-06-23T00:00:00 1899-12-30T15:45:00 2447 95 878 S12-1 T04-S03 Saturday 90 2012


2839 ENDO12L_S12-2 SYMPOSIUM SESSION: TRANSLATIONAL - BAT: Development [amp] Therapeutic Potential (3:45 PM - 5:15 PM) BAT as a Therapeutic Target Sven Enerbach G[ouml]teborg University, G[ouml]teborg, Sweden Disclosure Incomplete: SE 2012-06-23T16:15:00 Theater C 2012-06-23T00:00:00 1899-12-30T16:15:00 6046 95 879 S12-2 T04-S03 Saturday 91 2012


2840 ENDO12L_S12-3 SYMPOSIUM SESSION: TRANSLATIONAL - BAT: Development [amp] Therapeutic Potential (3:45 PM - 5:15 PM) Transcriptional Control of Brown Adipocyte Development and Function Patrick Seale University of Pennsylvania, Philadelphia, PA Brown fat cells are specialized to dissipate energy and can, therefore, counteract obesity. Fate mapping studies revealed that brown adipose and skeletal muscle cells originate from common or similar progenitors in the somitic mesoderm. In vitro and fate mapping studies suggest that muscle and brown fat cells diverge from a common progenitor in the somitic mesoderm early in development. The large zinc finger transcription factor, Prdm16 is a key driver of brown fat cell fate. Recent studies related to the role of Prdm16 and its closely related paralog, Evi1 in adipocyte development will be discussed.[br][br]Sources of Research Support: Diabetes Endocrinology Research Center (DK19525) and NIDDK/NIH (DK081605, OD007288).[br][br]Nothing to Disclose: PS 2012-06-23T16:45:00 Theater C 2012-06-23T00:00:00 1899-12-30T16:45:00 2430 95 880 S12-3 T04-S03 Saturday 92 2012


2841 ENDO12L_S13-1 SYMPOSIUM SESSION: BASIC - Cancer Stem Cells: Where Do We Start? (3:45 PM - 5:15 PM) Notch-Androgen Receptor Axis in Prostate Cancer Stem Cells Susan Kasper University of Cincinnati, Cincinnati, OH Disclosure Incomplete: SK 2012-06-23T15:45:00 Room 361 2012-06-23T00:00:00 1899-12-30T15:45:00 6022 96 881 S13-1 T02-S05 Saturday 94 2012


2842 ENDO12L_S13-2 SYMPOSIUM SESSION: BASIC - Cancer Stem Cells: Where Do We Start? (3:45 PM - 5:15 PM) Identification of Quiescent, Stem-Like Cells in the Distal Female Reproductive Tract Leendert J Blok Erasmus University, Rotterdam, Netherlands [bold]Background. [/bold]In fertile women, the endometrium undergoes regular cycles of tissue build-up and regression. It is likely that uterine stem cells are involved in this remarkable turnover. The main goal of the reported investigations was to identify (quiescent) endometrial stem cells in mouse by means of a pulse-chase approach to selectively earmark, prospectively isolate, and characterize label-retaining cells (LRCs).[br][bold]Methodology / Principal Findings. [/bold]To this aim, transgenic mice expressing the fusion protein histone 2B - GFP (H2B-GFP) in a Tet-inducible fashion were employed. After 7 days of doxycycline treatment (pulse), the doxycycline water was replaced by normal water (chase). Over time, dividing cells progressively loose the GFP signal after each mitosis, whereas quiescent (meaning infrequently dividing) cells will retain H2B-GFP expression. We evaluated H2B-GFP retaining cells at different chase time points and identified long-term ([underline][gt][/underline]12 weeks) LRCs in the distal oviduct. These LT-LRCs do not express estrogen receptor-alpha and a low level of progesterone receptors A and B (ER[sup]-[/sup] and PR[sup]-[/sup]), which is reminiscent of stem cells in other hormone-regulated organs such as the mammary gland. Furthermore, it was observed that FACSorted LT-LRCs are able to form spheroids capable of self-renewal and differentiation. Interestingly, under serum stimulation these spheroid cells can form glandular structures [italic]in vitro[/italic] which express markers of mature Mullerian epithelial cells.[br][bold]Conclusions / Significance. [/bold]In summary, it is shown that quiescent cells located in the distal oviduct have stem-like properties and can differentiate into distinct cell lineages specific of endometrium, and proximal and distal oviduct. Future lineage-tracing studies will be needed to elucidate the role played by these cells in homeostasis (for example during the menstrual cycle), tissue injury (after pregnancy, or perhaps induction of endometriosis) and cancer of the female reproductive tract (endometrial and ovarian cancer).[br][br]Nothing to Disclose: LJB 2012-06-23T16:15:00 Room 361 2012-06-23T00:00:00 1899-12-30T16:15:00 2401 96 882 S13-2 T02-S05 Saturday 95 2012


2843 ENDO12L_S13-3 SYMPOSIUM SESSION: BASIC - Cancer Stem Cells: Where Do We Start? (3:45 PM - 5:15 PM) EMT Programs [amp] Therapeutic Resistance of Breast Cancer Stem Cells Jeffrey Rosen Baylor College of Medicine, Houston, TX The claudin-low subtype is a recently identified rare molecular subtype of human breast cancer(1). Like basal-like tumors, these tumors are generally triple (ER, PR, HER2) negative and important therapeutically as there are currently no targeted agents directed at them. Claudin-low tumors express low levels of tight and adherens junction genes including claudin 3 and E-cadherin, and high levels of markers associated with epithelial-mesenchymal transition (EMT) including the EMT inducers Snail, Twist, Zeb1, and Zeb2. These tumors are also enriched in recently published signatures derived from human tumor-initiating cells (TIC) and a sorted population enriched for human mammary stem cells. miRNAs are also differentially expressed in claudin-low tumors including low expression of the miR-200 family - previously reported to regulate EMT and stemness(2). Currently, it is unknown whether the presence of the EMT phenotype is due to intrinsic genetic and epigenetic differences within a population of cells or a response to extrinsic factors emanating from the tumor microenvironment. However, the resulting intratumoral heterogeneity represents one of the major factors involved in tumor relapse and recurrence. Reactivation of the embryonic program known as epithelial-mesenchymal transition (EMT) during cancer progression promotes tumor invasion and metastasis. Induction of EMT in epithelial cells results in acquisition of molecular and functional traits of cancer stem cells (CSC), such as the ability to self-renew. Studies comparing paired human breast cancer core biopsies before and after chemotherapy demonstrated that CSCs were intrinsically chemoresistant(3). Moreover, the gene signature from these residual tumors identified the EMT/CSC enriched claudin-low and metaplastic subtypes(4). An independently derived core EMT interactome gene expression signature was also associated with these tumor subtypes(5). Furthermore, post-treatment residual tumors contained a higher fraction of claudin-low cells, also enriched with EMT/CSC properties, consistent with their therapeutic resistance(4).[br][br]1.Prat, A., et al (2010) Breast Cancer Res. 12:R68 PMID: 20813035. 2. Herschkowitz, J.I., et al. (2011) Proc. Natl. Acad Sci, USA, June 1. [Epub ahead of print]. PMID: 21633010. 3. Xiaoxian Li, Michael T. Lewis, Jian Huang, Carolina Gutierrez, C. Kent Osborne, Meng-Fen Wu, Susan G. Hilsenbeck, Gary C. Chamness, Helen Wong, Jeffrey Rosen, and Jenny C. Chang. (2008) J. Natl. Cancer Inst., 100:672-679. PMID: 18445819. 4. Creighton, C., et al. (2009) Proc. Natl. Acad. Sci,, USA, 106:13820-5. PMID: 19666588. 5. Taube, J.H., et al.(2010) Proc. Natl. Acad. Sci. USA, 107:15449-54, PMID: 20713713.[br]The author would like to acknowledge the following co-authors: Jeffrey M. Rosen[sup]1[/sup]; Jason I. Herschkowitz[sup]1[/sup]; Aleix Prat[sup]2[/sup]; Jana Knezevic[sup]1[/sup]; Sendurai A. Mani[sup]3[/sup]; Charles M. Perou[sup]2[/sup].[sup]1[/sup]Baylor College of Medicine, Houston, TX, [sup]2[/sup]University of North Carolina, Chapel Hill, NC, [sup]3[/sup]University of Texas MD Anderson Cancer Center, Houston, TX.[br][br]Sources of Research Support: This work is supported by grant RO1CA148761 from the National Cancer Institute.[br][br]Nothing to Disclose: JR 2012-06-23T16:45:00 Room 361 2012-06-23T00:00:00 1899-12-30T16:45:00 2405 96 883 S13-3 T02-S05 Saturday 96 2012


2844 ENDO12L_S14-1 SYMPOSIUM SESSION: TRANSLATIONAL - Development [amp] Function of Hypothalamic Circuitry (3:45 PM - 5:15 PM) Sex-Dependent Programming of Stress Pathways: Are We Wired before Birth? Tracy L Bale University of Pennsylvania, Philadelphia, PA Sex-biased neurodevelopmental disorders, including depression, anxiety, autism and schizophrenia, have been associated with maternal stress or perturbations experienced during pregnancy. The mechanisms and transgenerational programming of offspring outcomes through which stress contributes to disease development are not well understood, though likely involve a complex interaction between the maternal environment and effects on the placenta. We have identified a sensitive period of early gestation where maternal stress has sex-dependent epigenetic programming effects on offspring stress pathway neurodevelopment. Male offspring show increased stress sensitivity as adults in behavioral and physiological measures including tests assessing cognitive performance and stress coping strategies. These males also show physiological features of dysmasculinization including reduced testosterone levels, smaller testes, and a shorter anogenital distance suggesting a disruption in normal perinatal masculinization. Mechanistically, we have identified dramatic changes in the neonatal brain microRNA environment in response to early prenatal stress. Inhibition of testosterone aromatization by formestane in male neonates renders their miRNA expression pattern indistinguishable from females. Upstream of these effects, we have examined how early pregnancy stress impacts the developing brain via direct changes in placental programming. As the placenta is a sex-specific tissue, we identified a limited set of genes that consistently differ between the sexes across pregnancy, all of which are located on the X or Y chromosomes. Utilizing a proteomics approach, we found a candidate placental glycosylation enzyme and its biochemical target proteins that are significantly affected by maternal stress in a sex-dependent manner. Chromatin immunoprecipitation analyses support a direct mechanism whereby stress experience decreases histone association with this gene, decreasing its expression. As protein glycosylation typically competes with phosphorylation events, such broad changes are likely to yield important outcomes in placenta function and nutrient support of the developing fetus following stress. These results may provide critical insight into the mechanisms contributing to sex biased disease vulnerability to stress during pregnancy, and may provide novel biomarkers predictive of at-risk pregnancies.[br][br]Sources of Research Support: NIH Grants MH087597 and MH091258 awarded to TLB.[br][br]Nothing to Disclose: TLB 2012-06-23T15:45:00 Room 320 2012-06-23T00:00:00 1899-12-30T15:45:00 2527 97 884 S14-1 T06-S07 Saturday 98 2012


2845 ENDO12L_S14-2 SYMPOSIUM SESSION: TRANSLATIONAL - Development [amp] Function of Hypothalamic Circuitry (3:45 PM - 5:15 PM) Developmental Programming of Feeding Behavior in Non-Human Primates Elinor L Sullivan Oregon Health and Science University, Beaverton, OR Alterations in maternal nutrition during development have long term effects on offspring behavior and energy balance regulation. The goal of this study was to examine the consequences of maternal obesity and high-fat diet (HFD) consumption on the behavioral determinants of energy balance (feeding behavior, food preference and physical activity) and anxiety like and social behavior in juvenile nonhuman primates. Offspring from female Japanese macaques consuming either a low fat (13%) control diet or a HFD (35%) were examined. Preference for diets of differing fat and sugar content were examined. Physical activity was measured by an accelerometer attached to a collar. The Human Intruder test and novel object tests were used to assess stress and anxiety responses to a social threat or novel item. These tests were adapted from tests used in children and have been shown to reliably assess individual differences in primate stress response and anxiety. Social behavior was assessed in response to introduction to a novel peer and in normal social housing. In situ hybridization was used to assess the serotonin system in the dorsal raphe and cerebrospinal fluid. Female offspring from HFD consuming mothers exhibited increased anxiety in response to novel objects and male HFD offspring display increased aggression in response to a novel peer. In their normal environment, offspring from HFD mothers displayed an increase in behaviors associated with anxiety, aggression and fear. The serotonin system of HFD offspring was suppressed in the dorsal raphe and in the cerebrospinal fluid. HFD offspring exhibited increased preference for fat and sugar. They also exhibited decreased latency to eat novel diets and increased time eating high fat foods. Diet dependent changes in physical activity level were observed such that HFD offspring failed to display a compensatory increase in activity in response to HFD consumption. This study indicates that maternal HFD consumption causes suppression of the central serotonin system resulting in heightened anxiety and increased aggression in juvenile nonhuman primates. As a majority of pregnant women are overweight and consume a HFD, this study has important implications for the metal health status of future generations. Also, maternal over nutrition leads to long-term changes in energy balance regulation and food preference such that HFD offspring are at increased risk for weight gain and obesity.[br][br]Sources of Research Support: This work was supported by grants from the US National Institutes of Health (RO1DK079194, DK60685-S2, DK7919481, DK079194-S1, R00163, and 5UL1 RR24140-05).[br][br]Nothing to Disclose: ELS 2012-06-23T16:15:00 Room 320 2012-06-23T00:00:00 1899-12-30T16:15:00 2539 97 885 S14-2 T06-S07 Saturday 99 2012


2846 ENDO12L_S14-3 SYMPOSIUM SESSION: TRANSLATIONAL - Development [amp] Function of Hypothalamic Circuitry (3:45 PM - 5:15 PM) Optogenetic [amp] Pharmacogenetic Deconstruction of Hypothalamic Feeding Circuits Scott M Sternson Janelia Farm Research Campus, Ashburn, VA We are investigating neural control of the motivation to eat. We have developed cell type specific optogenetic and pharmacogenetic tools and we are using them to examine the causal relationship of electrical activity in specific neuronal cell types to hypothalamic neural circuit function and feeding behavior.[br][br]Nothing to Disclose: SMS 2012-06-23T16:45:00 Room 320 2012-06-23T00:00:00 1899-12-30T16:45:00 2426 97 886 S14-3 T06-S07 Saturday 100 2012


2847 ENDO12L_S15-1 SYMPOSIUM SESSION: TRANSLATIONAL - Hormonal Regulation of Inflammation [amp] Vascular Biology (3:45 PM - 5:15 PM) Insulin Signaling in CV Function James R Sowers University of Missouri - Columbia, Columbia, MO Two-thirds of American adults are overweight or obese, 75 million have hypertension and another 25 million have diabetes. The prevalence of overweight and obesity is even higher in our Veteran population. Decreased insulin sensitivity in cardiovascular tissue as well as in traditional targets of insulin metabolic signaling, such as skeletal muscle, is an underlying abnormality in obesity, hypertension, and type 2 diabetes. Data from our laboratory and others suggest that angiotensin II and aldosterone, in addition to excess nutrients, activate the evolutionarily conserved mTOR/S6K1 signaling pathway which, in turn, inhibits the metabolic signaling of insulin in cardiovascular and skeletal muscle tissue. In the cardiovascular system, insulin resistance in concert with excess nutrients and heightened tissue renin-angiotensin-aldosterone system (RAAS) activity may manifest as attenuated vasodilation, impaired myocardial glucose uptake and diastolic relaxation. Recent research examining the mechanisms by which heightened tissue RAAS and excess nutrients induce insulin resistance suggests that impaired insulin metabolic actions may be linked to excessive serine phosphorylation of insulin receptor substrate 1 (IRS-1). This critical insulin signaling/docking protein is normally tyrosine phosphorylation by the activated insulin receptor and serves to coordinate downstream signaling events by binding and activating effector proteins, such as PI3-K, leading to activation of Akt. In this context, phosphorylation of specific IRS-1 serine moieties can lead to impaired insulin metabolic signaling. Although several serine kinases have been shown to serine phosphorylate IRS-1, I will focus my review on the [bold]mTOR/S6K1 signaling pathway, which acts as a convergence point by which intake of excess nutrients and hormones, such as angiotensin II and aldosterone, converge.[/bold][br][br]Nothing to Disclose: JRS 2012-06-23T15:45:00 Room 351 2012-06-23T00:00:00 1899-12-30T15:45:00 2524 98 887 S15-1 T09-S06 Saturday 102 2012


2848 ENDO12L_S15-2 SYMPOSIUM SESSION: TRANSLATIONAL - Hormonal Regulation of Inflammation [amp] Vascular Biology (3:45 PM - 5:15 PM) Cerebrovascular Protection: An Estrogenic Role for Dihydrotestosterone during Inflammation and Ischemic Injury Rayna J Gonzales University of Arizona, Phoenix, AZ Dihydrotestosterone (DHT) may provide protective effects in the cerebrovasculature during pathophysiological conditions (such as ischemic stroke). During cerebral ischemia activation of NF[kappa]B or HIF-1[alpha] results in increased inflammation and oxidative stress due to increased transcription of pro-inflammatory mediators such as cytokines, chemokines, adhesion molecules and pro-inflammatory enzymes, such as cyclooxygenase-2 (COX-2). Our past studies have shown that DHT, independent of androgen receptor (AR) stimulation, decreases expression of COX-2 and HIF-1[alpha] during cytokine or hypoxia plus glucose deprivation exposure in human vascular smooth muscle (VSM) cells (2) and human brain microvascular endothelial cells. Additionally, we also demonstrate that chronic administration of DHT (400 pg/ml plasma) in vivo attenuates infarct size following focal ischemia in gonadectomized male rats (unpublished observation). Classically, DHT is regarded as a pure AR agonist; however, it can be endogenously metabolized to 5[alpha]-androstane-3[beta], 17[beta]-diol (3[beta]-diol), a compound recently shown to be a selective estrogen receptor (ER[beta]) agonist (1). In our current studies, we hypothesized that DHT[apos]s anti-inflammatory properties following interleukin-1[beta] (IL-1[beta]) stimulation are mediated through ER[beta]. Using primary human brain VSM cells (HBVSMC) we verified the presence of steroid receptors (AR, ER[alpha], and ER[beta]) and the enzymes (3[beta]-HSD and 17[beta]-HSD) necessary to synthesize 3[beta]-diol using PCR. Next, we demonstrated that DHT (10 nM) attenuated IL-1[beta] induced COX-2 and HIF-1[alpha] protein expression and that the AR antagonist (bicalutamide) did not alter this response. Conversely, both the non-selective ER antagonist ICI 182,780 and the selective ER[beta] antagonist PHTPP inhibited the effect of DHT, suggesting that DHT actions are ER[beta]-mediated. DHT also attenuated IL-1[beta]-induced ROS generation. In both HBVSMC and in rat mesenteric arteries, 3[beta]-diol, similar to DHT, reduced cytokine-induced COX-2 levels. In conclusion, our data indicate that the androgen, DHT, working through ER[beta] may provide protection against vascular oxidative stress under pathophysiological conditions, in part by decreasing pro-inflammatory mediators and subsequent ROS production; and thereby perhaps preserving cerebrovascular function.[br][br](1) Weihua Z et al., Proc Natl Acad Sci U S A. 2002; 99:13589-94. (2) Zuloaga K and Gonzales R, American Journal of Physiology Heart & Circulatory Physiology, 2011; 301(5):H1882-90. Additional Contributing Authors: Kristen Zuloaga, PhD and Robert Handa, PhD.[br][br]Sources of Research Support: American Heart Association Scientist Development Grant awarded to RJG; University of Arizona Sarver Heart Center Doris Griswold Award for Novel Research in the Area of Cardiovascular Disease and Medicine awarded to KLZ and the Madson Family Gift Fund awarded to RJG.[br][br]Nothing to Disclose: RJG 2012-06-23T16:15:00 Room 351 2012-06-23T00:00:00 1899-12-30T16:15:00 2518 98 888 S15-2 T09-S06 Saturday 103 2012


2849 ENDO12L_S15-3 SYMPOSIUM SESSION: TRANSLATIONAL - Hormonal Regulation of Inflammation [amp] Vascular Biology (3:45 PM - 5:15 PM) Nuclear Receptors in Vascular Inflammation [amp] Atherosclerosis Willa Ann Hsueh The Methodist Hospital, Houston, TX Nuclear receptors lie at the crossroads of inflammation and metabolism, and many have been implicated as therapeutic targets for obesity and its inflammation-driven complications. Thiazolidinediones (TZDs), insulin sensitizing PPARgamma ligands, are anti-inflammatory and inhibit progression of vascular lesions in middle-aged, low-density lipoprotein cholesterol receptor deficient (Ldlr-/-) mice fed 3 months of Western diet; a model of obesity, metabolic syndrome and markedly accelerated atherosclerosis. TZDs also have profound effects to promote atherosclerosis regression in this model, possibly through stimulation of the reverse cholesterol transporters ABCA1 and ABCG1 and through inhibition of inflammation. However, TZDs have side effects that severely restrict their use in diabetes. Recently, a new class of PPARgamma ligands has been developed that inhibit cdk5 phosphorylation of PPARgamma and promote insulin sensitization but have attenuated effects on adipogenesis, suggesting they may have a better side effect profile. However, these agents have little anti-inflammatory capability. With our collaborators, Drs. Ivan Pitta (Instituto Nacional de Ciencia e Tecnologia para Inovacao Farmaceutica) and Francisco Neves (Universidade de Brasilia), we have characterized a novel class of PPARgamma ligands, GQs, which are insulin sensitizing, inhibit cdk5 phosphorylation, have attenuated effects on adipogenesis, but possess strong anti-inflammatory activity. These agents particularly target interleukin-1alpha and -1beta, which are implicated in the pathogenesis of obesity-associated insulin resistance, type 2 diabetes, and atherosclerosis. GQs inhibit accelerated atherosclerosis in Ldlr-/- mice and decrease inflammatory responses in adipocytes and adipose immune cells. GQs may be useful therapy for diabetes, obesity and cardiovascular disease.[br][br]Sources of Research Support: NIH/NIDDK Grant 1R24DK087723-01 awarded to WAH.[br][br]Disclosure Incomplete: WAH 2012-06-23T16:45:00 Room 351 2012-06-23T00:00:00 1899-12-30T16:45:00 2544 98 889 S15-3 T09-S06 Saturday 104 2012


2850 ENDO12L_S16-1 SYMPOSIUM SESSION: BASIC - Nuclear Receptors in Development [amp] Physiology (3:45 PM - 5:15 PM) Functional Role of RIP140 in the Development of Metabolic Diseases Li-Na Wei University of Minnesota Medical School, Minneapolis, MN Receptor-interacting protein 140 (RIP140) is a co-regulator for various transcription factors and nuclear receptors, and is expressed mainly in metabolic tissues. Extensive post-translational modifications occur on RIP140, which affects its sub-cellular distribution and functionality (1). Its biological activity in whole animals is most profound for the progression of T2DM, as established in examining gene knockout mice. Our recent studies have established two types of biological activities for RIP140, one in the nucleus and another in the cytoplasm. Previously we have established its principal action, which is to negatively regulate hormone target gene expression through recruiting various chromatin remodeling components and enzymes (2, 3). More recent years we have begun to uncover its novel activity in the cytoplasm. This presentation will discuss these recent findings, which are most relevant to the progression of metabolic diseases, including its regulatory activity for GLUT4 vesicle trafficking (4), adiponectin vesicle secretion (5), ET1 secretion (6) and lipolysis (7) in adipocytes, and its contribution to inflammation in macrophages thereby modulating endotoxin tolerance status in animals (8). Finally, I will also discuss the effect of diets on the level and types of protein modifications of RIP140 (9), which ultimately contribute to its biological activities, and the metabolic status of whole animals.[br][br]1. Mostaqul Huq MD, Gupta P and Wei L-N (2008) Post-translation Modification of Nuclear Co-repressor RIP140: potential targets of therapeutics. Curr. Med. Chem. 15: 386-92. 2. Park SW, Huang, WH, Persaud, SD and L-N Wei (2009) RIP140 in thyroid hormone-repression and chromatin remodeling of Crabp1 gene during adipocyte differentiation. Nucl Acid Res. 37: 7085-94. 3. Shawna D. Persaud, Wei-Hong Huang, Sung Wook Park, and Li-Na Wei (2011) Gene repressive activity of RIP140 through direct interaction with CDK8. Mol. Endocrinol. 25: 1689-98. 4. Ping-Chih Ho, Yi-Wei Lin, Yao-Chen Tsui, Pawan Gupta, and Li-Na Wei (2009) A negative regulatory pathway of GLUT4 trafficking in adipocyte: new function of RIP140 in the cytoplasm via AS160. Cell Metabolism 10: 516-523. 5. Ping-Chih Ho and Li-Na Wei (2012) Negative regulation of adiponectin secretion by receptor interacting protein 140 (RIP140). Cell. Signal. 24:71-76. 6. Ping-Chih Ho, Yao-Chen Tsui, Yi-Wei Lin, Shawna D. Persaud, Li-Na Wei (2012) Endothelin 1 promotes cytoplasmic accumulation of RIP140 through a ETA-PLC[beta]-PKC[epsilon] pathway. Mol. Cell Endocrinol. 351:176-83. 7. Ho PC, Chuang YS, Hung CH, Wei LN. (2011) Cytoplasmic receptor-interacting protein 140 (RIP140) interacts with perilipin to regulate lipolysis. Cell. Signal. 23:1396-403. 8. P-C Ho, Y-C Tsui, X Feng, DR Greaves and L-N Wei (2012) NF-kB-mediated RIP140 degradation regulates inflammatory response and contributes to endotoxin tolerance. Nature Immunology, in press. 9. Ho, P-C, Chang, K-C, Chuang, Y-S and Wei, L-N (2011) Cholesterol regulation of receptor interacting protein 140 via microRNA-33 in inflammatory cytokine production. FASEB J. 25: 1758-66.[br][br]Sources of Research Support: This work is supported by NIH grants DK60521, DK54733, and K02DA13926, as well as The Distinguished McKnight University Professorship of the University of Minnesota.[br][br]Nothing to Disclose: L-NW 2012-06-23T15:45:00 Room 342 ABDE 2012-06-23T00:00:00 1899-12-30T15:45:00 2391 99 890 S16-1 T08-S02 Saturday 106 2012


2851 ENDO12L_S16-2 SYMPOSIUM SESSION: BASIC - Nuclear Receptors in Development [amp] Physiology (3:45 PM - 5:15 PM) Therapeutic Targeting of NR4A Orphan Nuclear Receptors for Treatment of Acute Myeloid Leukemias Orla M Conneely Baylor College of Medicine, Houston, TX Acute myeloid leukemias (AML) represent a heterogeneous group of blood malignancies with poor prognosis. The orphan nuclear receptors (ONRs), NR4A1 and NR4A3, are potent tumor suppressors of AML. They are silenced in all human AML patients regardless of cytogenetic background. Combined NR4A1/3 deletion in mice leads to extremely rapid early postnatal AML development due to disruption of hematopoietic stem cell (HSC) homeostasis and the emergence of a transformed radioresistant leukemia initiating cells (LICs). To address the therapeutic potential of NR4A reactivation in AML, we have addressed the cellular and molecular consequences of Nr4A1/3 rescue in human AML cells. We show that acute rescue of NR4A1 or NR4A3 in human AML cells inhibits their proliferation and reprograms a subset of gene signatures that distinguish all primary human LICs from normal HSCs regardless of cytogenetics. Through integration of NR4A regulated gene signatures with chemical genomics we have identified novel chemical inducers of NR4As that reduce AML leukemogenicity, reprogram a subset of common LIC gene signatures, and surprisingly, drive lymphoid trans-lineage priming of human AML blasts.[br]Together, our results identify NR4A1/3 as novel therapeutic targets in AML and provide a general strategy for discovering chemical modulators of ONR activity.[br][br]Nothing to Disclose: OMC 2012-06-23T16:15:00 Room 342 ABDE 2012-06-23T00:00:00 1899-12-30T16:15:00 2508 99 891 S16-2 T08-S02 Saturday 107 2012


2852 ENDO12L_S16-3 SYMPOSIUM SESSION: BASIC - Nuclear Receptors in Development [amp] Physiology (3:45 PM - 5:15 PM) COUP-TFII Controls Prostate Cancer Progression [amp] Metastasis Ming-Jer Tsai Baylor College of Medicine, Houston, TX Hyperactivation of the PI3-kinase/AKT pathway as a result of PTEN mutation is a driving force for prostate cancer. However, the precise mechanism underlying how indolent tumors with PTEN alternations acquire metastatic potential remains largely undefined. Here we show that COUP-TFII, a member of the nuclear receptor superfamily, serves as an important regulator to override the TGF-b-dependent checkpoint for PTEN null indolent tumors. Over-expression of COUP-TFII in the mouse prostate epithelium cooperates with PTEN deletion to exacerbate prostate tumor progression and develop an aggressive metastasis-prone tumor. Mechanistically, COUP-TFII is shown to inhibit TGF-b signaling through hindering Smad4 transcription activity. The biological significance of COUP-TFII is substantiated by patient sample analysis, in which COUP-TFII expression is tightly correlated with disease progression and tumor recurrence, while it is inversely correlated with TGF-b signaling. These findings reveal that the destruction of the TGF-b dependent-barrier by COUP-TFII is important for the progression of PTEN mutant prostate cancer to a life-threatening disease and raises a potential new drug target for intervention of metastatic prostate cancer.[br][br]Sources of Research Support: This work is supported by NIH grants, HD17379 and DK45641.[br][br]Nothing to Disclose: M-JT 2012-06-23T16:45:00 Room 342 ABDE 2012-06-23T00:00:00 1899-12-30T16:45:00 2448 99 892 S16-3 T08-S02 Saturday 108 2012


2853 ENDO12L_S17-1 SYMPOSIUM SESSION: CLINICAL - Pediatric Implications of Reproductive Endocrine Technologies (3:45 PM - 5:15 PM) Pediatric Outcomes of IVF Therapy Paul Leslie Hofman University of Auckland, Auckland, New Zealand Since the birth of the first in vitro fertilization (IVF) baby in 1978, this technique has grown in popularity, accounting for 1[ndash]3% of births in Western countries with more than 2 million babies born worldwide. IVF is a relatively new technology and largely first generational. Follow-up studies are limited and most relate to pregnancy and neonatal outcome with relatively few looking at long term follow up in later childhood. IVF children have an increased risk of major malformation detected in infancy and there is a reported increased risk of imprinting disorders including the overgrowth disorder Beckwith-Wiedemann syndrome and the neurodevelopmental disorder, Angelman syndrome. Cross sectional studies in mid childhood have demonstrated increased stature with a corresponding increase in growth factors and beneficial alteration in lipid profiles. Interestingly there is evidence of sexual dimorphism with girls born using IVF being relatively taller than boys. This has raised the possibility that there may be subtle alterations in gene expression. The cause for these changes has generated many hypotheses. These include a bias due to selection of larger embryos, ovarian hyperstimulation, the nutrient media used during blastocyst culture and endocrine disrupters such as bisphenol A from the use of plastic containers during culture. We have investigated several of these hypotheses in a number of related studies. These include investigating ovarian hyperstimulation using children born to mothers using clomiphene, examining outcomes in children born from frozen embryos pregnancies and investigating epigenetic alterations in children born following IVF. This talk will summarize this data on longer term follow up in humans demonstrating that there appear to be subtle but clear differences in the phenotype of children born following assisted reproductive technologies.[br][br]Nothing to Disclose: PLH 2012-06-23T15:45:00 Room 310 2012-06-23T00:00:00 1899-12-30T15:45:00 2404 100 893 S17-1 T07-S04 Saturday 110 2012


744 ENDO12L_SAT-727 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) The FGFR4 Transmembrane Polymorphism Signals through Distinct Stat3 Modifications To Regulate Corticotroph Hormone Feedback and Cell Growth Tae Tateno, Toru Tateno, Lei Zheng, Maw Maw Hlaing, Katsuhiko Yoshimoto, Shozo Yamada, Sylvia Asa, Shereen Ezzat University of Toronto, Toronto, Canada; University of Toronto, Toronto, Canada; University of Tokushima Graduate School, Tokyo, Japan; Toranomon Hospital, Tokyo, Japan Background: Fibroblast growth factor receptor 4 (FGFR4) is a member of a family of four transmembrane receptors with ligand-induced tyrosine kinase activity. FGFRs have been implicated in pituitary tumorigenesis. We recently demonstrated that a single nucleotide polymorphism (SNP) substituting an arginine (R) for glycine (G) in the FGFR4 transmembrane domain alters cell signaling, resulting in increased cell proliferation and growth hormone production in mammosomatotroph cells (1). Here, we examined the differential properties of the two FGFR4 isotypes on cell signaling, hormone production and cellular growth in corticotroph cells.[br]Methods and Results: Mouse AtT20 corticotroph tumor cells were stably transfected with constructs encoding V5-tagged full length FGFR4 Gly388 cDNA or FGFR4 Arg388 cDNA. The levels of FGFR4, POMC expression, and phosphorylation of STAT3 (pS-STAT3 and pY-STAT3), STAT1 and STAT5 were examined before and after dexamethasone treatment. FGFR4-G388 cells had higher activated pY-STAT3, which resulted in increased Pro-opiomelanocortin (POMC) expression and resistance to dexamethasone. This effect was recapitulated by introducing a constitutively active pY-STAT3 mutant. In contrast, FGFR4-R388 corticotrophs supported pS-STAT3 signaling to stimulate colony formation without impact on dexamethasone-mediated POMC inhibition. Clinical data from 36 patients (G/G:14, G/R:17, R/R:5) with Cushing[apos]s disease revealed that those homozygous for the R/R allele were more likely to harbor a pituitary macroadenoma. In contrast, patients harboring the G/G allele showed microadenomas or no lesions on MR imaging.[br]Conclusion: These data support a link between the FGFR4 SNP, corticotroph hormone production, and resistance to dexamethasone feedback.[br][br]PLoS Genet. 2011;7(12):e1002400. Epub 2011 Dec 8.[br][br]Nothing to Disclose: TT, TT, LZ, MMH, KY, SY, SA, SE 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 375 76 805 SAT-727 PO22-01 Saturday 744 2012


745 ENDO12L_SAT-728 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Testicular Orphan Nuclear Receptor 4 (TR4) [mdash] Novel Regulator of Corticotroph Function and Growth Li Du, Marvin Bergsneider, Leili Mirsadraei, William H Yong, Anthony P Heaney David Geffen School of Medicine, University of California, Los Angeles, CA; David Geffen School of Medicine, University of California, Los Angeles, CA; David Geffen School of Medicine, University of California, Los Angeles, CA Cushing[apos]s disease (CD) is a life-threatening neuroendocrine disorder, due to excess corticotroph tumor ACTH and adrenal steroid secretion and lacks safe and effective medical therapy. We have observed abundant immunocytochemical nuclear expression of the orphan nuclear receptor, testicular receptor 4 (TR4) in human pituitary tumors in comparison to low cytoplasmic expression in normal pituitary tissues. TR4 over-expression in murine corticotroph tumor AtT20 cells resulted in a dose-dependant increase in POMC promoter luciferase activity, POMC mRNA and protein expression. In additional studies, siRNA directed knockdown of TR4 expression in AtT20 and primary cultures of human corticotroph tumors suppressed POMC transcription and expression. Chromatin immunoprecipitaion (ChIP) assay demonstrated that TR4 binds the POMC promoter at a consensus direct repeat-1 (DR1, AGGTCA) locus. Co-immunoprecipitation studies demonstrate that TR4 binds the glucocorticoid receptor (GR) to regulate glucocorticoid/GR- mediated negative feedback on POMC transcription. Overexpression of TR4 increased whereas TR4 knockdown suppressed murine and human corticotroph proliferation [italic]in vitro [/italic]and murine corticotroph tumor growth[italic] in vivo[/italic].[br]These findings demonstrate that TR4 is a novel potent regulator of corticotroph tumor POMC transcription and cell growth and demonstrate that TR4 interacts with the glucocorticoid receptor (GR). Further characterization of TR4-mediated POMC regulation may provide important insights into the mechanisms involved in regulation of corticotroph ACTH secretion and glucocorticoid/GR- mediated feedback on corticotroph function and growth.[br][br]Sources of Research Support: Jonnson Cancer Center.[br][br]Nothing to Disclose: LD, MB, LM, WHY, APH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2028 76 806 SAT-728 PO22-01 Saturday 745 2012


746 ENDO12L_SAT-729 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) PRKCD Is a Target of miR-26a in ACTH Pituitary Adenoma Erica Gentilin, Federico Tagliati, Teresa Gagliano, Daniela Mole, Ettore degli Uberti, Maria Chiara Zatelli University of Ferrara, Section of Endocrinology, Ferrara, Italy microRNAs (miRNAs) have several physiological functions, but have been also implicated in human neoplastic initiation and progression. We previously demonstrated that 30 miRNAs are differentially expressed in normal human pituitary as compared to pituitary adenomas. However, the most of miRNAs target genes remain unknown, hindering the understanding of the miRNAs contribute to pituitary tumorigenesis.[br]The aims of this study were to: (1) validate a murine ACTH-secreting pituitary adenoma cell line as a possible model to study pituitary miRNA deregulation; (2) validate and investigate the role of potential targets of differentially expressed miRNAs. We analysed the murine AtT-20/D16v-F2 cell line, deriving from a murine ACTH-secreting pituitary adenoma, and normal mouse pituitary for the expression pattern of 11 miRNA, which expression was found to be different in human pituitary adenomas vs. normal pituitary. Our results showed a partial agreement (50%) between expression trend of these miRNAs in human and mouse. In particular, we found that miR-26a has overlapping expression patterns in humans and mice, being up-regulated in adenomas vs. normal pituitary. Our results confirm that the 3[apos] untranslated region of PRKCD, a miR-26a putative target gene, is a functional target of this miRNA and provide evidence, by Real Time PCR, that this target is translationally suppressed. PRKCD, a member of the PKC subfamily, is dynamically involved in cell apoptosis in a specific stimulus manner. We observed that miR-26a inhibition led to a decrease in cell viability without increasing caspase 3/7 activity. These results indicate that miR-26a is overexpressed in human ACTH pituitary adenoma and can control cell viability in AtT-20/D16v-F2 cell line, by reducing the PRKCD expression, playing an important role in pituitary adenoma development. Our study provides new insights into potential contribution of these RNAs to pituitary neoplastic transformation and suggests that miR-26a might be a possible target for therapeutic strategies.[br][br]Nothing to Disclose: EG, FT, TG, DM, EdU, MCZ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 581 76 807 SAT-729 PO22-01 Saturday 746 2012


747 ENDO12L_SAT-730 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Different Entitities of Corticotroph Adenomas on the Basis of Prohormone Convertase Expression and the Relationship with Dopamine D2 Receptor Expression Rob van der Pas, Steven Lamberts, Federico Gatto, Alberto Pereira, Max Kros, Sjoerd van Duinen, Richard Feelders, Leo Hofland Erasmus Medical Center, Rotterdam, Netherlands; Leiden University Medical Center, Leiden, Netherlands; Erasmus Medical Center, Rotterdam, Netherlands; Leiden University Medical Center, Leiden, Netherlands [bold]Introduction[/bold][br]Pro-opiomelanocortin (POMC) is the precursor molecule of ACTH. In the pituitary gland, POMC is cleaved by prohormone convertases (PC) 1 and 2. PC1 is believed to be expressed in both the anterior and intermediate pituitary lobe and cleaves POMC into ACTH and [beta]-lipotrophin (LPH). PC2 further processes these POMC products to [alpha]-MSH, CLIP, [gamma]-LPH and [beta][ndash]endorphin. PC2 expression is restricted to the intermediate lobe, which is believed to regress soon after birth and, therefore, to be absent in adults.[br]Cushing[apos]s disease (CD) is caused by excess production of ACTH by a pituitary corticotroph tumor. It is generally accepted that these tumors originate from corticotroph cells in the anterior pituitary. However, observations that tumors localized in the region between the anterior and posterior pituitary lobe are harder to remove by transsphenoidal surgery and are more often responsive to dopamine 2 receptor (D2R) agonists led to the hypothesis that CD can not only originate from corticotroph adenomas of the anterior lobe, but also from an adenoma or hyperplasia of ACTH-producing cells of the intermediate zone.[br][bold]Methods[/bold][br]Using RT-PCR and immunohistochemistry, mRNA and protein expression levels of PC1, PC2 and D2R were examined in pituitary tissue of 45 patients with CD that underwent surgery. These data were correlated with radiological findings and histology.[br][bold]Results[/bold][br]Either at mRNA or protein level there was no significant overall correlation between PC1 or PC2 and D2R expression. PC1 or PC2 expression was not correlated with UFC excretion either.[br]However, when the PC2 immunoreactivity score (IRS) was divided in tertiles, it was found that, compared to the lowest tertiles, the group of adenomas with the highest PC2 protein expression had a lower prevalence of macroadenomas on MRI (4/17 vs. 16/27; p[lt]0.05) and a higher prevalence of adenomatous hyperplasia at histological examination (8/16 vs. 2/26; p[lt]0.01). The group with the highest PC2 tertile tended to have higher D2R IRS (6.53 [plusmn] 3.17 vs. 4.81 [plusmn] 2.60; p=0.06).[br][bold]Conclusion[/bold][br]Compared to ACTH-secreting pituitary tumors with low PC2 immunostaining, tumors with high PC2 staining are more often classified as adenomatous hyperplasia of ACTH producing cells rather than as adenoma. This suggests that CD can be caused by an adenoma or hyperplasia of ACTH-producing cells of the intermediate zone of the pituitary gland. Moreover, D2R protein expression appears to be higher in adenomas with high PC2 protein expression.[br][br]Nothing to Disclose: RvdP, SL, FG, AP, MK, SvD, RF, LH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 912 76 808 SAT-730 PO22-01 Saturday 747 2012


748 ENDO12L_SAT-731 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) The Role of Mediators of Cell Invasiveness, Motility and Migration in the Pathogenesis of Silent Corticotroph Adenomas Ozgur Mete, Gelareh Zadeh, Fred Gentili, Shereen Ezzat, Sylvia L Asa University Health Network, Toronto, Canada; University Health Network, Toronto, Canada; University Health Network, Toronto, Canada [bold]Background:[/bold] Silent corticotroph adenomas (SCA) represent a distinct histopathological variant of clinically nonfunctioning pituitary adenoma. It has been speculated that they are more aggressive than other non-functioning adenomas (NFA). We investigated the expression of some markers related to cell invasiveness, cell motility and migration, and compared the data with the more common NFA that usually have features of gonadotroph differentiation.[br][bold]Methods:[/bold] We studied 12 SCA and compared them with a larger series of usual NFA. Semiquantitative immunohistochemistry was performed to localize [beta]1-integrin, osteopontin, and MMP-1 as cytoplasmic, membranous, or mixed cytoplasmic-membranous. The results are recorded as positive when [gt]5% of tumors cells are positive and the extent of positivity for each antibody is assigned to one of four following scores: 5-25% (score 1), 26-50% (score 2), 51-75% (score 3) and 76-100% (score 4). The mean score is used for comparison.[br][bold]Results:[/bold] [bold][beta]1-integrin[/bold]: All cases were positive for [beta]1-integrin. No significant difference was present between SCA and usual NFA. Extent of [beta]1-integrin expression was significantly different within subgroups of SCA (p[lt]0.05); mean extent of staining was higher in type II SCAs (3) than type I SCA (2). [bold]MMP-1: [/bold]MMP-1 immunoreactivity was observed in 83% of SCA and 93% of NFA. Mean extent of staining is higher in NFA (3), followed by type II (2.5) and type I SCA (1.5). Interestingly, similar correlation is observed with the FGFR4 expression levels and MMP-1 score of tumor groups (NFA: 93%, type II SCA: 71%, type I SCA: 25%). [bold]Osteopontin:[/bold] While osteopontin immunoreactivity was observed in all SCA, only 60% of NFA were positive. Extent of osteopontin expression was significantly different between SCA and NFA (p[lt]0.05); mean extent of staining was higher in SCA (type II: 3.6, type I: 3.5) than NFA (2.5).[br][bold]Conclusions[/bold]: Type II SCA is biologically more aggressive than type I SCA. While osteopontin seems to play a major role in the invasiveness of SCAs, a high MMP-1 score is characteristic of NFAs. Moreover, high FGFR4 levels of NFAs seem to be responsible for the induction of MMP-1 expression in NFAs.[br][br]Nothing to Disclose: OM, GZ, FG, SE, SLA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1020 76 809 SAT-731 PO22-01 Saturday 748 2012


749 ENDO12L_SAT-732 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Putative Role of [italic]miR-145[/italic] Underexpression in [italic]gsp+ [/italic]Somatotropinomas Ana Rosa Quidute, Fernanda Borchers Coeli, Paula Lamparelli Elias, Margaret Castro School of Medicine of Rebeir[atilde]o Preto - University of S[atilde]o Paulo, Rebeir[atilde]o Preto, Brazil [bold]Introduction:[/bold]Sporadic somatotropinomas are highly associated with [italic]GNAS[/italic] gene mutation (5-40%) whereas [italic]AIP gene [/italic]mutation is only found in 3%. Recently, miRNAs have been involved in pituitary tumorigenesis. [bold]Objectives:[/bold] To identify the [italic]GNAS[/italic] and [italic]AIP[/italic] mutations; to evaluate the expression of[italic] GNAS[/italic] gene and a panel of miRNAs in mutated [italic](gsp[bold]+[/bold])[/italic] and non-mutated [italic](gsp[bold]-[/bold])[/italic] somatotropinomas; to evaluate the association of gene/miRNA expression with clinical features. [bold]Materials and methods:[/bold][italic]GNAS[/italic] gene and miRNAs expression were evaluated by qPCR [italic](2[sup]-[/sup][/italic][italic][sup][Delta][Delta]Ct[/sup]) [/italic]in[italic] 26 [/italic]sporadic somatotropinomas and 7 normal pituitaries (NP[italic])[/italic]. Tumor size was evaluated by CT/RMN using [italic]Hardy[/italic] score. Baseline IGF-I was expressed in percentage of upper limit of normal range (%ULNR) age- and sex-adjusted. [bold]Results:[/bold] We found [italic]9[/italic] out of 26 [italic](35%) [/italic]gsp+ (5 [italic]p.R201C,[/italic] 2 [italic]p.R201H and 2 p.Q227L) [/italic]and [italic]AIP[/italic] mutation (p.R304X) in one ([italic]4%) [/italic]gsp-tumor. The mean age of patients at diagnosis was similar: [italic]gsp+[/italic] (39.0[plusmn]11) and [italic]gsp-[/italic] (43.5[plusmn]9) groups as well the mean pre-operative GH (62.4[plusmn]128 [italic]vs [/italic]39.9[plusmn]48.31;[micro]g/l; IFMA) and IGF-1 (435[plusmn]230 [italic]vs[/italic] 557[plusmn]238; [italic]%ULNR[/italic]), respectively[italic].[/italic] The Hardy Score distribution in [italic]gsp+[/italic] and [italic]gsp-[/italic]tumors were similar. There was no differential [italic]GNAS[/italic] expression between somatotropinomas and NP as well as between gsp+ and gsp- tumors. There was no differential expression in[italic] gsp+ and [/italic]gsp- tumors regarding miR-15a, miR-23a, miR-23b, miR-24-2, miR-150, miR-141, miR-16, miR-143, miR-21, let-7a. However[italic],[/italic] [italic]miR-145 [/italic]was underexpressed in gsp+ tumors (p=0.03). Tumors classified as Hardy I/II showed higher [italic]GNAS[/italic] expression than III/IV (p=0.008); however there was no difference in miR-145 expression in both groups. Among the gsp+ group, 7 patients were followed up; all these patients (100%) had controlled disease (2 by transsphenoid surgery, 4 on somatostatin agonist and 1 on somatostatin and dopaminergic agonists). However, only 9 out of 17 (53%)[italic] gsp-[/italic]tumors had controlled disease even with combined therapy. In conclusion, the prevalence of [italic]GNAS[/italic] and [italic]AIP[/italic] mutations in sporadic somatotropinomas was similar to worldwide. Our study reinforces previous data suggesting that patients with [italic]gsp+[/italic] tumors have better response to somatostatin agonists. The [italic]miR-145[/italic] underexpression in [italic]gsp+ [/italic]somatotropinomas could be a consequence of the gsp oncogene deregulation or would suggest a putative role of this miRNA in controlling directly or indirectly the expression of transcription factors that mediate cAMP pathway.[br][br]Sources of Research Support: FAPESP 2007/58365-3.[br][br]Nothing to Disclose: ARQ, FBC, PLE, MC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2080 76 810 SAT-732 PO22-01 Saturday 749 2012


750 ENDO12L_SAT-733 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Clinical and Genomic Profiling of Growth Hormone Tumor Subtypes To Identify Mechanisms of Disease Pathogenesis, Aggressivness, and Response to Therapy Katja Kiseljak-Vassiliades, Shibana Shafi, Mei Xu, Tzu L Phang, Bette K Kleinschmidt-DeMasters, Kevin O Lillehei, Janice M Kerr, Margaret E Wierman University of Colorado Denver, Aurora, CO; Veterans Affairs Medical Center Denver, Denver, CO; University of Colorado Denver, Aurora, CO; University of Colorado Denver, Aurora, CO; University of Colorado Denver, Aurora, CO; University of Colorado Denver, Aurora, CO Growth hormone (GH) tumors are associated with metabolic, respiratory, cardio and cerebrovascular complications leading to shortened life span with persistent or recurrent disease. Histologically, GH tumors are characterized into two sub-types: densely-granulated (DG) and sparsely-granulated (SG) based on immunohistochemical (IHC) features and cytokeratin distribution. Literature suggests that SG tumors are more common in the young ([lt]50 yrs), with larger tumors and lower GH/IGF-1 levels than DG tumors. SG tumors are thought to be less responsive to somatostatin analogs than DG tumors, but the underlying mechanisms are poorly defined. In a cohort of patients where molecular profiling was performed, SG patients (N=10) were not significantly younger than DG (N=11), 43[plusmn]5 and 51[plusmn]3yrs, respectively (p=0.20). Tumor size, basal IGF-1 and GH also did not differ between SG and DG patients. However, 3 months after transsphenoidal resection, IGF-1 levels were lower in DG compared to SG tumors, 266[plusmn]76 and 693[plusmn]131ng/mL, respectively (p=0.017). All SG (N=10) patients, but only 9% (N=1/11) of DG patients required medical therapy following the surgery. While somastostatin analogues resulted in 45.02% reduction of IGF-1 levels, normalization was not achieved in any of SG patients at 1yr follow up. To identify new differential mechanisms involved in tumorigenesis and progression of GH tumor subtypes, Affymetrix exon expression microarray was performed. Biostatistical analysis with an ANOVA model was used to examine differentially expressed transcripts with false discovery rate of 5%. 672 deregulated genes with 2-fold change were identified. 47.7% of differentially expressed genes were upregulated in SG compared to DG tumors. Significantly (2-fold) deregulated genes were further explored using the Ingenuity Pathway Analysis software identifying numerous pathways that are significantly different (Fisher Exact Test, p[lt]0.05) between SG vs DG tumors, including cell-to-cell interaction (p=1.9x10[sup]-5[/sup]) and cellular assembly and organization (p=8.9x10[sup]-6[/sup]) pathways. These pathways will be further explored as potential disease predictors and underling molecular mechanisms in GH tumorigenesis. In summary, pathologic GH tumor subtyping is useful to predict more aggressive GH tumors. Future studies are needed to identify additional predictors of GH tumor behavior and response to medical therapy, and uncovering pathways in GH tumorigenesis to develop novel therapeutic targets.[br][br]Sources of Research Support: This work was funded by NIH K08-DK069511 to JMK and VA Merit Review to MEW, UCD Cancer Center and Genomics Core.[br][br]Nothing to Disclose: KK-V, SS, MX, TLP, BKK-D, KOL, JMK, MEW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1983 76 811 SAT-733 PO22-01 Saturday 750 2012


751 ENDO12L_SAT-734 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) GH Receptor and IGFBP3 Polymorphisms in Acromegaly: Correlation with Clinical and Laboratory Features and Response to Therapy Raquel S Jallad, Ericka B Trarbach, T Chile, Felipe HG Duarte, Alexander AL Jorge, Marcello D Bronstein School of Medicine of the University of S[atilde]o Paulo, S[atilde]o Paulo, Brazil; School of Medicine of the University of S[atilde]o Paulo, S[atilde]o Paulo, Brazil; School of Medicine of the University of S[atilde]o Paulo, S[atilde]o Paulo, Brazil Background: Polymorphisms d3GHR and -202 A/C IGFBP3 have been associated with clinical presentation, biochemical measurements and response to therapies in acromegaly. Objective: To evaluate the presence of these polymorphisms in acromegalics and their influence on clinical and laboratorial characteristics of patients, at diagnosis and after treatment. Patients and Methods: A total of 123 patients (76 men, 47 women, age range 26[ndash]88 years) with acromegaly were evaluated at diagnosis(basal) and after treatment. GHR-exon 3 polymorphisms were assessed by multiplex PCR. -202 A/C IGFBP3 polymorphisms were genotyped by RFLP. Both polymorphisms presented genotype distribution consistent with the Hardy-Weinberg equilibrium. The relationship among log basal GH, IGF-1 Z, age and of genotypes GHR-exon3 and -202 A/C IGFBP3 was evaluated by multiple linear regression. A p value of [lt]0.05 was considered statistically significant. Results: The distribution of patients in the three different GHR-exon 3 genotypes was 45% for fl/fl, 46 % for fl/d3 and 10 % for d3/d3. The frequencies of -202 A/C IGFBP3 genotypes were 31% for C/C, 51% for A/C and 17% for A/A. In the evaluation after treatment, comparing with A *genotype, patients with C/C genotype presented higher levels of basal GH(P[lt]0.001), basal IGF-I(P=0.014) and IGF-1 Z (P=0.005). There were no significant influence of genotypes GHR-exon3 and -202 A/C IGFBP3 on clinical (age, sex and body mass index) and laboratorial characteristics(GH, IGF-I, IGF-1 Z and IGFBP3 values) of patients at diagnosis and after treatment. The only variable that affected basal GH was age (p = 0.022) and the only variable that influenced IGF-1 Z was log basal GH (p= 0.044). Polymorphisms d3GHR and -202 A/C IGFBP3 had no influence on the levels of GH and IGF-1 Z, at the diagnosis and after treatment. The results were expressed as mean [plusmn] SD and range. IGF-I was expressed as SDS. Discussion and Conclusion: Unlike reports from the literature (1-4), the present data did not shown any significant influence of genotypes GHR-exon3 and -202 A/C IGFBP3 on clinical and laboratorial features in acromegalics patients at diagnosis and after treatment.[br][br]1-Kamenicky P, Dos Santos C, Espinosa C, Salenave S, Galland F, Le Bouc Y, Maison P, Bougn[egrave]res P, Chanson P 2009 D3 growth hormone receptor polymorphism is not associated with IGF-1 levels in untreated acromegaly. Eur J Endocrinol 161:231[ndash]235. 2-Schmid C, Krayenbuehl PA, Bernays RL, Zwimpfer C, Maly FE, Wiesli P 2007 Growth hormone (GH) receptor isoform in acromegaly: lower concentrations of GH but not insulin-like growth factor-1 in patients with a genomic deletion of exon 3 in the GH receptor gene. Clin Chem 53:1484[ndash]1488. 3-Bianchi A, Giustina A, Cimino V, Pola R, Angelini F, Pontecorvi A, De Marinis L 2009 Influence of growth hormone receptor d3 and full-length isoforms on biochemical treatment outcomes in acromegaly. J Clin Endocrinol Metab 94:2015[ndash]2022. 4- Mercado M, Gonz[aacute]lez B, Sandoval C, Esquenazi Y, Mier F, Vargas G, de los Monteros AL, Sosa E 2008 Clinical and biochemical impact of the d3 growth hormone receptor genotype in acromegaly. J Clin Endocrinol Metab 93:3411[ndash]3415.[br][br]Sources of Research Support: Financial support from FAPESP.[br][br]Nothing to Disclose: RSJ, EBT, TC, FHGD, AALJ, MDB 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1934 76 812 SAT-734 PO22-01 Saturday 751 2012


752 ENDO12L_SAT-735 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Serial Analysis of Gene Expression in Pituitary Tumors: Wnt Canonical and Non-Canonical Pathways Are Not Involved in Pituitary Tumorigenesis Leandro Machado Colli, Beatriz MC Paixao, Fernando C Amaral, Deison S Lima, Wilson Silva, Ayrton C Moreira, Sonir R Antonini, Ricardo ZN Vencio, Margaret Castro School of Medicine of Rebeir[atilde]o Preto - University of S[atilde]o Paulo, Rebeir[atilde]o Preto, Brazil; School of Medicine of Rebeir[atilde]o Preto - University of S[atilde]o Paulo, Rebeir[atilde]o Preto, Brazil; School of Medicine of Rebeir[atilde]o Preto - University of S[atilde]o Paulo, Rebeir[atilde]o Preto, Brazil; Faculty of Philosophy, Sciences and Letter of Ribeir[atilde]o Preto, Rebeir[atilde]o Preto, Brazil Introduction: Serial analysis of gene expression (SAGE) technique has been largely used to measure global gene expression. Wnt pathway genes are involved in general tumorigenesis; the role of the canonical Wnt pathway in pituitary tumorigenesis is still controversial. Two Wnt non-canonical pathways have never been studied in pituitary tumors. The aim of this study was to evaluate the differential gene expression profile by SAGE and validate the expression of canonical and non-canonical Wnt pathway genes in different subtypes of pituitary tumors. Materials and methods: Four SAGE cDNA libraries were constructed using a pool of mRNA obtained from 5 GH-, 2 ACTH-secreting, and 4 non secreting pituitary tumors (NS), and 3 normal pituitaries from patients who had accidental death, using I-SAGE kit (Invitrogen). We used a Bayesian model (SAGEci) for SAGE differential gene expression ratio to carry out statistical analyses. In addition, we evaluated in 21 GH-, 20 ACTH-secreting and 22 NS pituitary tumors, and in 6 normal pituitaries the expression of 31 Wnt pathway genes: Wnt Canonical pathway (WNT11, WNT4, WNT5A, DKK3, sFRP1, CTNNB1, APC, AXIN1, GSK3[beta], AKT1, CDH1, TCF7, MAPK8, and NFAT5), WNT/Calcium pathway (PLCB1, CAMK2A, PRKCA, and CHP), Planar cell polarity pathway (PRICKLE, VANGL1, DVL-1, DVL-2, DVL-3, PTK7, DAAM1, and RHOA), and targets genes (MYB, MYC, WISP2, SPRY1, and TP53), and endogenous controls (GUSB, TBP, and PGK1). Gene expression was calculated by QPCR software. Results: We identified 46,688 genes in all four SAGE libraries. Comparing pituitary tumor SAGE libraries to normal pituitary library, we found 446 genes downregulated and 14 upregulated. ACTH SAGE library had 1 gene exclusively upregulated and 4 downregulated compared to other SAGE libraries. GH library had 2 genes exclusively upregulated and 12 downregulated whereas NS library had 2 genes exclusively upregulated and 13 downregulated compared to other libraries. There was no differential gene expression in Wnt canonical and non canonical pathways between pituitary tumors and normal pituitaries, but DKK3 in ACTH-secreting (-2.5x; p=0.01) and NS (-5.0x; p=0.02), and TCF7 in NS (-6.6x; p=0.02). Conclusions: SAGE identified several genes differently expressed, which can be further studied in order evaluate their involvement in each specific pituitary tumor lineages. Our data provide evidences that Wnt canonical and non-canonical pathways seem not to be involved in pituitary tumorigenesis.[br][br]Nothing to Disclose: LMC, BMCP, FCA, DSL, WS, ACM, SRA, RZNV, MC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1661 76 813 SAT-735 PO22-01 Saturday 752 2012


753 ENDO12L_SAT-736 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Sox 2 Expression in Human Pituitary Adenomas: An Immunohistochemical Study Marius Raica, Anca Cimpean, Cristina Capatina, Mihail Coculescu Victor Babes University of Medicine and Pharmacy, Timisoara, Romania; Carol Davila University of Medicine and Pharmacy, Bucharest, Romania Sox2 is a marker of embryonic stem cell pluripotency and plays a role in normal pituitary development but also in the progression of tumors. However, Sox2 expression in pituitary adenomas and its possible correlation with clinicopathologic characteristics have not been investigated so far.[br][bold]Aim.[/bold] To evaluate by immunohistochemistry the expression of SOX2 protein in pituitary adenomas.[br][bold]Subjects and methods.[/bold] Pituitary adenoma samples prelevated during neurosurgery from 34 patients diagnosed and treated at the C.I. Parhon National Institute of Endocrinology in Romania were analyzed by immunohistochemistry for SOX2 expression in the primary tumor samples by the avidin-biotin-HRPA method using monoclonal primary anti-SOX2 antibodies. From the 34 tumors 13 were GH-secreting, 10 were prolactinomas and 10 clinically non-functioning adenomas.[br][bold]Results.[/bold] SOX2 positive expression was detected in 16 patients (47.05 % of cases): 8 cases with acromegaly, 6 prolactinomas and 2 non-functioning adenomas and did not show an association with tumor size or extension at diagnosis.[br]GH-secreting tumors were immunopositive for SOX 2 in 57.14% of cases, prolactinomas in 60% and non-functioning pituitary adenomas in only 20% of cases (p=0.044 in non-functioning compared to functioning adenomas). The 8 GH-secreting tumors expressing SOX 2 expressed only GH in 4 cases, GH and PRL in 3 cases and GH, PRL and FSH in another case. The 6 prolactinomas immunopositive for SOX 2 protein expressed only prolactin (4 cases) or associating synthesis of GH (without clinical expression) in 1 case and LH (in another). The 2 non-functioning adenomas expressing SOX2 were null-cell adenomas.[br]The 18 SOX2 negative tumors were 6 GH secreting (expressing only GH in 4 cases, GH-PRL in one case and GH-PRL-TSH in another), 4 prolactinomas (with monohormonal secretion) and 8 non-functioning adenomas (7 null cell tumors and one with silent GH expression).[br][bold]Conclusion.[/bold] SOX2 positive expression is frequent in pituitary adenomas, especially in secreting tumors.[br][br]Nothing to Disclose: MR, AC, CC, MC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1930 76 814 SAT-736 PO22-01 Saturday 753 2012


754 ENDO12L_SAT-737 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Evaluation of the Expression of Vesicular Protein (SNARE) Isoforms in Human Pituitary Adenomas Giampaolo Trivellin, Edwin A Garcia, Michael Powell, Joan Grieve, Ian Sabin, Ghassan Alusi, Louis Pobereskin, Babak Shariati, Federico Roncaroli, Nigel Mendoza, Ashley B Grossman, Elaine A Harper, Marta Korbonits Queen Mary University of London, London, UK; University College London Hospitals, London, UK; Barts and The Royal London Hospital, London, UK; Derriford Hospital, Plymouth, UK; Imperial College London, London, UK; University of Oxford, Oxford, UK; Syntaxin Ltd, Abingdon, UK Background: Botulinum neurotoxin (BoNT) inhibits muscle function by interfering with neurotransmitter release from secretory vesicles. The mechanism underlying this effect involves cleavage of SNARE proteins which are required for vesicle docking at the plasma membrane. The ability of BoNT serotypes to cleave SNARE proteins including VAMP1, VAMP2, VAMP3, syntaxin1 and SNAP-25, and inhibit secretion is being exploited for therapeutic purposes by Syntaxin Ltd with their [apos]targeted secretion inhibitor[apos] (TSI) technology. A specific TSI, SXN101959, is believed to have therapeutic utility in treating acromegaly because it specifically targets pituitary somatotrophs and cleaves VAMP proteins to bring about an inhibition of GH secretion. The expression of SNARE isoforms has not been previously documented in detail in human pituitary.[br]Aims: To characterise the expression of different SNARE isoforms in human pituitary adenomas and normal pituitaries.[br]Methods: Specimens from 66 patients undergoing transphenoidal surgery as a therapeutic option for pituitary adenomas were studied: nineteen acromegalics intervened for the first time, two acromegalics intervened for a second time, five with Cushing[apos]s disease, 39 with non-functioning pituitary adenomas (NFPAs,) and one with a pituitary prolactinoma. The specimens were dispersed and cells profiled for vesicle SNAREs (VAMP1, VAMP2 and VAMP3) and membrane-bound SNAREs (syntaxin1, SNAP-23, and SNAP-25) by immunofluorescence and/or Western blotting. In addition, mRNA expression levels of SNAREs were measured in 14 somatotrophinomas and 15 NFPAs by RT-qPCR. Six normal pituitaries from autopsy were included as controls.[br]Results: VAMP2 and VAMP3 were found in normal pituitary and pituitary adenomas and in some cases VAMP1 was also shown to be present at the mRNA level. 72% of the 66 adenomas were found to be positive for VAMP2 and/or VAMP3 by immunofluorescence and/or Western blotting. Syntaxin1 was detected in all pituitary samples at both the mRNA and protein level while SNAP-23 was dominant in pituitary adenomas especially in somatotrophinomas where co-localisation with GH was observed. SNAP-25 was rarely documented in adenomas but was shown to be present in the normal pituitary.[br]Conclusions: The present data suggest that the VAMP2[amp]3, SNAP-23 and syntaxin1 are the predominant SNARE-proteins in somatotroph adenomas. These data provide the basis for the therapeutic use of Syntaxin Ltd[apos]s SXN101959 in the treatment of acromegaly.[br][br]Nothing to Disclose: GT, EAG, MP, JG, IS, GA, LP, BS, FR, NM, ABG, EAH, MK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2325 76 815 SAT-737 PO22-01 Saturday 754 2012


755 ENDO12L_SAT-738 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) The Role of Estrogen Pathway on Aryl Hydrocarbon Receptor Interacting Protein(Aip) [ndash] Related Pituitary Tumorigenesis Suk Kyoung Kim, Young-Suk Choi, Cheol Ryong Ku, Eun Jig Lee Yonsei University College of Medicine, Seoul, Korea; Yonsei University College of Medicine, Seoul, Korea; Yonsei University College of Medicine, Seoul, Korea Germline mutations in the aryl hydrocarbon receptor interacting protein (Aip) gene are frequently accompanied with familial pituitary adenoma. Aip serves as a tumor suppressor gene and one of the main target molecules is aryl hydrocarbon receptor (AhR). Although some studies reported the crosstalk between aryl hydrocarbon receptor (AhR) and estrogen receptor (ER), there is no report evaluating the relationship between estrogen pathways and[italic] Aip[/italic] in pituitary tumoriogenesis.[br]For in vitro study, we utilized the GH3 cell line which is one of the rat pituitary tumor cell lines. Loss of function in [italic]Aip[/italic] was achieved with siRNA for [italic]Aip[/italic]. MTT assay and [italic]Reverse Transcriptase[/italic]-PCR were performed. For in vivo study, we generated somatotroph-specific [italic]Aip[/italic] knock out (sAIPKO) mice using Cre-loxp strategy under C57/BL6 background.[br]The size of pituitary gland from mice was different depending on the distinction of sex and the existence of [italic]Aip[/italic]. In sAIPKO mice, pituitary glands of female were significantly larger than those of male one. Furthermore, female sAIPKO mice had significantly larger pituitary gland than female control C57/BL6 mice. In MTT assay, [italic]Aip[/italic]-deficient GH3 cells treated with estrogen showed increased proliferation. To investigate the relationship between altered estrogen signaling and increased proliferation, we performed RT-PCR on [italic]Aip[/italic] deficient GH3 cells with or without treatment of estrogen. Loss of [italic]Aip[/italic] in GH3 cells increased the expression of estrogen receptor-related genes, including estrogen receptor-[alpha] and estrogen receptor-[beta].[br]Our data suggest that estrogen influence the proliferation of somatotroph depending on the expression level of [italic]Aip[/italic]. Further investigation with sAIPKO mice might be helpful understanding the mechanisms evolving the relationships between estrogen and [italic]Aip[/italic].[br][br](1) Elina Hellovaara et al., AJP 2009; 175: 2501. (2) Anniina Raitila et al., AJP 2010; 177: 1969.[br][br]Nothing to Disclose: SKK, Y-SC, CRK, EJL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1471 76 816 SAT-738 PO22-01 Saturday 755 2012


756 ENDO12L_SAT-739 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Aromatase Expression in Human Pituitary Adenomas Paulo Henrique Gonzales, Maria Beatriz Kohek, Adriana Vial Roehe, Nelson Pires Ferreira, Julia FS Pereira-Lima, Carolina GS Leaes, Miriam da Costa Oliveira Universidade Federal de Ci[ecirc]ncias da Sa[uacute]de de Porto Alegre (UFCSPA), Porto Alegre, Brazil; ISCMPA/Universidade Federal de Ci[ecirc]ncias da Sa[uacute]de de Porto Alegre (UFCSPA), Porto Alegre, Brazil Introduction: Several studies point to a relevant role of estrogen in pituitary tumorigenesis. The nuclear estrogen receptors [alpha] and [beta], are present in all normal adeno-pituitary cells, with studies showing its expression in pituitary adenomas. Estrogen stimulates the expression of the pituitary tumor-transforming gene (PTTG), which exhibits marked expression in malignant cell lines and pituitary tumors. The aromatase, enzyme responsible for converting androgens into estrogens, has a crucial role in the estrogen biosynthesis in various normal, hyperplastic and neoplastic tissues. In humans, there is only study evaluating the expression of aromatase in normal pituitary tissue, revealing its presence in both sexes, while the presence of aromatase in human pituitary adenomas has not yet been investigated. Objectives: Evaluate the expression of aromatase in human pituitary adenomas, through immunohistochemistry (IH), and quantitative real time polymerase chain reaction (qRT-PCR). Material and Methods: We assessed clinical data and surgical material of 65 patients undergoing hypophysectomy with anatomopathological diagnosis of pituitary adenoma. The material removed was subjected to the IH and qRT-PCR techniques for the assessment of the expression of aromatase. Results: The sample consisted of 35 women (53.9%) and 30 men (46.2%). Of the 65 patients, 56 (86.2%) had macroadenomas and 9 (13.9%) microadenomas. Immunohistochemical data showed 23 patients (35.4%) with non-functioning pituitary adenomas, 19 (29.2%) with somatotrophic adenomas, 12 (18.5%) with lactotrophic adenomas and 11 (16.9%) with corticotrophic adenomas. The IH evaluation, showed loss of expression of aromatase in all samples, except one. Quantification by qRT-PCR showed loss of gene expression in 36/43 cases. The concurrent assessment of the material by the two techniques showed loss of gene expression and protein content of the enzyme in 30/37 cases (concordance rate of 81.1%). Conclusion: In human pituitary adenomas occurs loss of expression of aromatase, regardless of the sex of the patients, hormonal subtype and tumor size. The findings suggest that the effects of estrogen in pituitary tumorigenesis are not related to the increase of their concentration in these neoplasms by local production of the enzyme.[br][br]Nothing to Disclose: PHG, MBK, AVR, NPF, JFSP-L, CGSL, MdCO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 434 76 817 SAT-739 PO22-01 Saturday 756 2012


757 ENDO12L_SAT-740 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) SXN101959 Selectively Inhibits GH Pulsatility in Cynomolgus Monkeys Alberto Martinez, Robert M Gill, Kenneth Gould, Aimee Cossins, Helen Ludlow, Ferdinand Roelfsema, Caroline Whately-Smith, Richard Jones Syntaxin Ltd, Abingdon, UK; Covance Laboratories, Inc, Greenfield, IN; LUMC, Leiden, Netherlands; Whately-Smith Ltd, King[apos]s Langley, UK [underline]Introduction:[/underline] SXN101959 is being developed as a treatment for acromegaly with a novel mechanism of action. In acromegalics, pituitary somatotroph adenomas secrete GH autonomously resulting in high levels of circulating GH. SXN101959 is a targeted secretion inhibitor (TSI) which targets the pituitary somatotrophs of rats, primates and man. Following receptor activation and internalization the endopeptidase is released into the cytoplasm and cleaves the target SNARE protein (VAMP) which inhibits the release of GH. SXN101959 and the predecessor SXN101742, have been shown to inhibit the pulsatile release of GH in rats. This study aimed to assess the magnitude and duration of GH pulsatility inhibition after a single IV bolus administration of SXN101959 in cynomolgus monkeys.[br][underline]Methods[/underline]: 16 na[iuml]ve cynomolgus monkeys were instrumented with femoral artery access. Animals were randomised into three groups, vehicle (n=6), 0.1mg/kg SXN101959 (n=5) and 0.3mg/kg SXN101959 (n=5). All blood sampling was conducted using an automated blood sampling system (Dilab). GH and PRL sampling occurred at 15 min intervals between 7:00pm and 3:45am (next morning) on days -1, 4, 14 and 28. Cortisol, Insulin, C-peptide, TRH, FSH and IGF-1 were also sampled on days throughout the study. Pharmacokinetic measurements of SXN101959 via ELISA were made at 0, 1, 2, 4, 8 and 24hrs after dosing.[br][underline]Results[/underline]: GH pulsatility was ablated in animals treated with a single bolus IV injection of 0.1mg/kg and 0.3mg/kg SXN101959 on day 4 after treatment. Animals treated with 0.1mg/kg SXN101959 showed recovery of pulsatility by day 14 while 0.3 mg/kg dosed animals remained inhibited for a longer duration through day 14 with residual activity 28 days after treatment. Cortisol, TSH, FSH, insulin, glucose and IGF-1 remained unchanged. GH AUC levels were unchanged (SXN101959 vs vehicle).[br][underline]Conclusions[/underline]: GH pulsatility inhibition was achieved by day 4 post dosing in all treated animals. The duration of GH secretion inhibition was longer with the increasing doses of SXN101959. Despite the successful inhibition of GH pulsatility no change was observed in AUC of GH which may explain the unchanged IGF-1 levels. This data suggests that IGF-1 in normal primates is driven by GH basal levels. In acromegalics the role of pulsatility remains controversial. The compound appears to be specific to the somatotroph since none of the other pituitary hormone levels were perturbed.[br][br]Disclosures: AM: Employee, SYNTAXIN. RJ: Employee, SYNTAXIN. Nothing to Disclose: RMG, KG, AC, HL, FR, CW-S 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 439 76 818 SAT-740 PO22-01 Saturday 757 2012


758 ENDO12L_SAT-741 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) PK Model Approach to Understanding the Efficacy of SXN101959 in Rats Alberto Martinez, Anne Wilson, Elaine Anne Harper, Richard Jones, Jan Freijer Syntaxin, Abingdon, UK; CHDR, Leiden, Netherlands [underline]Introduction:[/underline] SXN101959 is being developed for the treatment of acromegaly. Pituitary adenoma somatotrophs are the source of the abnormally high GH levels. SXN101959 is a novel targeted secretion inhibitor (TSI) designed to specifically target the somatotroph. Once SXN101959 is internalized into an endosome the endopeptidase is delivered into the cytoplasm which cleaves VAMP, the target SNARE protein, thereby inhibiting the release of GH. Previous reports have focused on bolus administration of SXN101959 in rats. The goal of the study was to use a population PK model derived from those studies to predict the SXN101959 PK profile in rat plasma after infusion and explore the effect on IGF-1 plasma levels.[br][underline]Methods[/underline]: PK model generated a number of treatment options combining bolus and infusion administration. Six groups of animals (8 SD male rats/group) were treated: group 1-vehicle, group 2-0.7mg/kg IV bolus, group 3-0.7mg/kg IV infusion (4 hrs), group 4-0.3mg/kg IV bolus + 0.4mg/kg IV infusion (4hrs), group 5-0.3mg/kg IV infusion (4hrs) and group 6-0.13mg/kg IV bolus+0.17mg/kg infusion (4 hrs). IGF-1 was sampled on days 1, 3, 5, 9, 12, 15, 18, 21 and IGF-1 levels determined (alpha-LISA, Perkin Elmer). Plasma levels of SXN101959 were assessed by ELISA 0, 1, 2, 4, 8, 12, 24, 48, 72 and 96 hours after dosing.[br][underline]Results[/underline]: PK profiles generated closely resembled the model predictions used for the experimental design. IGF-1 mean plasma levels co-related to SXN10959 total exposure (AUC[sub]0-[/sub][yen]) regardless of the route. Increasing the exposure to SXN101959 led to an increase in the magnitude of the inhibition of IGF-1 plasma levels as well as the duration of the inhibition. Groups receiving 0.7mg/kg had a maximal inhibition of 48% IGF-1 compared to control animals while those given 0.3mg/kg reduced plasma IGF-1 by 41%. 0.3mg/kg treated groups IGF-1 plasma levels returned to baseline by day 21 while 0.7mg/kg groups IGF-1 plasma levels remained inhibited by 22%.[br][underline]Conclusions[/underline]: Inhibition of IGF-1 plasma levels by SXN101959 was related to exposure levels regardless of the route of administration. The data indicates an exposure effect on both the magnitude and duration of the inhibition after a single administration of SXN101959. The study suggests that using infusion approaches can improve the overall inhibition of IGF-1 levels while reducing high peak concentrations of SXN101959.[br][br]Disclosures: AM: Employee, SYNTAXIN. RJ: Employee, SYNTAXIN. Nothing to Disclose: AW, EAH, JF 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 342 76 819 SAT-741 PO22-01 Saturday 758 2012


759 ENDO12L_SAT-742 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Sleep Apnea in Acromegaly: Pathogenetic Factors and Long-Term Follow-Up Cinzia Castellani, Giuseppe Francia, Marcello Ferrari, Elena Viva, Maria Vittoria Davi Internal Medicine D, Verona, Italy; ENT, Verona, Italy [bold]Background: [/bold][br]Sleep apnea syndrome (SAS) is a common disease in acromegaly and it can persist during remission in up to 58% of the patients. Data regarding long term outcome of SAS in acromegalic patients are lacking. Moreover it is still unknown which component, either craniofacial deformation or soft tissue hypertrophy of the palate and upper airways, may play the major role in the pathogenesis of this complication.[br][bold]Aim[/bold][br]To assess the presence of SAS in a series of acromegalic patients including active and controlled patients and to perform follow up in patients with SAS after biochemical control or during long-term remission. Moreover to evaluate site, degree and possible cause of upper airways obstruction by MRI and fiberoptic nasopharyngoscopy with the Muller maneuver (FNMM).[br][bold]Patients and method[/bold][br]Polysomnography was performed in 58 acromegalic patients: 33 active and 25 controlled and was repeated in 25 patients with SAS of whom 16 after achieving biochemical control and 9 after long term remission (mean 6,6 years SD [plusmn] 3,2). In 29 patients morphological study of the upper airways by MRI and FNMM was carried out.[br][bold]Results[/bold][br]The prevalence of SAS was 64% in active and 52% in controlled patients. Among 16 active patients 8 (50%) showed SAS improvement and 2 (12,5%) recovered after biochemical control, whereas in 9 out of 13 (69,2%) controlled patients SAS persisted. Uvula alone or with tongue base was the main site of obstruction assessed by FNMM in 90% of patients. Uvula diameters correlated with the severity of airways narrowing at FNMM and tongue measure with the severity of the AHI.[br][bold]Conclusions[/bold][br]SAS can improve after biochemical control of acromegaly, but can persist even after long term follow up despite recovery from acromegaly. Hypertrophy of soft palate and tongue is relevant factor responsible for occurrence and severity of SAS.[br][br]Nothing to Disclose: CC, GF, MF, EV, MVD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1574 76 820 SAT-742 PO22-01 Saturday 759 2012


760 ENDO12L_SAT-743 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Activity Control of Acromegaly without Pharmacological Treatment during Pregnancy: A Prospective Study Monike L Dias, Nina R Musolino, Monica R Gadelha, Cesar L Boguszewsky, Jose G Vieira, Julio Abucham Federal University of S[atilde]o Paulo, S[atilde]o Paulo, Brazil; University of S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; Federal University of Parana, Curitiba, Brazil [bold]Introduction: [/bold]Acromegalic gestations are mostly uneventful, but the absence of prospective series may be leading to underestion of real prevalence of clinical signs and symptoms. Growth hormone (GH) during pregnancy has to be measured by assays free from placental peptides[apos] interference, but assays crossreactivities are rare descriptions on retrospective cases yet published. We show the first prospective series of pregnancies in acromegaly with clinical, biochemical and radiological assessment and GH measurements by a GH assay resistant to pregnancy-related interferences.[br][bold]Methods: [/bold]Inclusion criteria were: (1) previous diagnosis of acromegaly and a pituitary adenoma; (2) active disease; (3) GH and IGF-I measurements within six months before, during pregnancy and after delivery using the same assays. Exclusion criterion was previous radiotherapy. No drugs were given after pregnancies were diagnosed. GH was measured by a modified in-house IFMA assay [sup]1[/sup], validated in pregnancy by dilution tests, and IGF-I by Immulite 2000[reg]. Sixty two normal pregnant women (17-39 years) were used as controls for GH and IGF-I levels.[br][bold]Results: [/bold]Nine pregnancies in six acromegalic patients (24-37 y) were consecutive and prospectively followed by endocrinologists between 2006 and 2012. All had previous transesphenoidal surgery. No patient developed hypertension or anterior pituitary deficiencies, either before or during pregnancy. Two patients developed mild gestational diabetes.No patient had visual complains or tumor growth during pregnancy. All deliveries occurred at term and all newborns were healthy and had normal height and weight. IGF-I levels in acromegalic patients were relatively controlled before pregnancy (range: 237-583 ng/ml, median 354; or 0,75 [ndash] 2,05 ULN), showed no significant change among trimesters ([italic]P= [/italic]0,93, range: 156 [ndash] 556 ng/ml, median 376 for first trimester, range: 127- 649 ng/ml, median 312 for 2[sup]nd[/sup] trimester, and range: 120-536 ng/ml, median 284 for 3[sup]rd[/sup] trimester) and no statistical difference from controls at 3[sup]rd[/sup] trimester. All patients had a return of IGF-I levels to pre-treatment levels, suggesting a transient control in pregnancy. GH levels remained stable throughout pregnancy in 3 of 4 pregnancies assessed.[br][bold]Conclusions: [/bold]Drug withdrawal in acromegaly during pregnancy may be safe in previously controlled patients. Tumor size, GH and IGF-I levels remain stable throughout gestation.[br][br]1.Vieira, JGH; Lombardi, MT; Nishida, S. Monoclonal antibody-based immunoenzymometric assay for human growth hormone. Brazilian J Med Biol Res 1990; 23:293-296.[br][br]Disclosures: MRG: Principal Investigator, Novartis Pharmaceuticals. Nothing to Disclose: MLD, NRM, CLB, JGV, JA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2378 76 821 SAT-743 PO22-01 Saturday 760 2012


761 ENDO12L_SAT-744 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Long-Term Remission of Acromegaly after Adjunctive Pharmacological Treatment with Octreotide and/or Cabergoline Walid Chacra, Alessandra Casagrande, Debora Nazato, Julio Abucham Escola Paulista de Medicina, Universidade Federal de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Introduction: Both somatostatin analogs and dopaminergic agonists, alone or in combination, have long been used to control pituitary tumor growth and serum levels of growth hormone and IGF-1 in acromegalic patients, most often after unsuccessful pituitary surgery. Although these drugs are able to control 30-60% of patients, disease remission after drug withdrawal is seldom attained (1,2,3).[br]Patients and Methods: In a retrospective survey of 39 acromegalic patients treated by surgery followed by octreotide and/or cabergoline and no radiotherapy, we identified 5 patients (all female) that remained controlled after drug withdrawal for at least 4 months (normal IGF-I for age and sex). All these patients had random serum GH [lt]2.0 [micro]g/l (range: 0.1 - 1.8 [micro]g/l, mean; 0.68 [micro]g/l). Previous pharmacological treatment had been stepwise and guided by serum IGF-I (starting with octreotide LAR 20 mg and followed sequentially, as necessary, by one or more steps: octreotide 30 mg I.M./month, addition of cabergoline 3,5 mg P.O./week, interruption of octreotide, reintroduction of octreotide). At diagnosis, 4 were macroadenomas. Residual pituitary masses were visible in only two patients at post-surgical MRI, but in none when drugs were withdrawn after 24-84 months of treatment (mean: 5 years). After drug withdrawal, patients were followed clinically and biochemically with serum GH (IRMA or ICLA) and IGF-I measurements (Immulite[reg] 2000, Siemens) at approximately 3-month intervals.[br]Results: Two patients treated for 24 and 72 months with octreotide alone and octreotide plus cabergoline remained clinically and biochemically controlled after 51 and 24 months of complete drug withdrawal, respectively. Three patients, treated for 60-84 months with octreotide plus cabergoline followed by cabergoline alone (2 patients) and octreotide alone, relapsed after 6, 10 and 24 months of drug withdrawal.[br]Conclusion: Acromegalic patients with hormonal control and no visible pituitary mass under pharmacological treatment for at least 24 months frequently sustain long-term disease remission after drug withdrawal.[br][br](1) Ramirez C, Vargas G, Gonzalez B,et al.,Discontinuation of octreotide LAR after long term, successful treatment of patients with acromegaly: is it worth trying? European Journal of Endocrinology 2012 166 21[ndash]26. (2)Verhelst JA, Abrams PJ, Abs R. Remission of acromegaly following long-turn therapy with cabergoline: report of two cases. Pituitary 2008 11 103-107. (3)Ronchi CL, Rizzo E, Lania AG, et al.,Preliminary data on biochemical remission of acromegaly after somatostatin analogs withdrawal. European Journal of Endocrinology 2008 158 19-25.[br][br]Nothing to Disclose: WC, AC, DN, JA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2210 76 822 SAT-744 PO22-01 Saturday 761 2012


762 ENDO12L_SAT-745 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Evaluation of Colonoscopic Procedure and Frequency of Colon Pathologies in Acromegaly Patients Erkan Caglar, Esra Hatipoglu, Mutlu Niyazoglu, Pinar Kadioglu Cerrahpasa Medical School, University of Istanbul, Istanbul, Turkey; Cerrahpasa Medical School, University of Istanbul, Istanbul, Turkey Objective: Since an increase in the frequency of colon pathologies (e.g. polyps) was reported in acromegaly patients, monitorization of patients with colonoscopy have become an important follow-up tool. Therefore, we investigated both the practicability and quality of colonoscopic examination in acromegaly patients and the frequency of colon pathologies in our center.[br]Materials and Methods: In our study, colonoscopies (and endoscopic procedure parameters) of 49 acromegaly patients (25 newly diagnosed, 24 with biochemical cure) and 73 age- and gender-matched nonacromegalic patients with irritable bowel syndrome were evaluated prospectively.[br]Results: Gender distribution (male/female) in acromegaly group was 24/25 and in control group (CG) was 27/46 (p=0.690). Mean age of the acromegaly patients and CG were 44.73[plusmn] 11.64 years and 43.90[plusmn] 10.73 years, respectively (p= 0.188). Cecum arrival rate of the acromegaly group and the CG were 87.7% (43/49) and 98.6% (72/73) respectively (p=0.011). Median cecum insertion time in acromegaly patients and in the CG was 330 seconds [IQR 240-520] and 230 seconds [IQR 190 -320], respectively (p=0.005). Bowel preparation was suboptimal (stool residue in lumen or liquid stool) in 14 (28.5%) of the acromegaly patients and in two (2.8%) of the patients in the CG (p[lt]0.001). Diverticulosis was detected in none of the patients with acromegaly (0%) and 3 (4.1%) of the CG patients (p=0.168). Colon polyps were observed in 14 (28.5%) of the acromegaly patients and only 5 (6.8%) of the CG cases (p=0.001).[br]Conclusion: Higher suboptimal bowel preparation rates and longer cecum insertion times in acromegaly group denote the difficulties of colonoscopic examination in cases with acromegaly. With all this, prolonged and more detailed preparation before colonoscopy may be necessary in this group. In the current study frequency of colonic diverticula was not higher in acromegaly patients, contrary to what was previously reported (1).[br][br]1. Wassenaar MJ, Cazemier M, Biermasz NR, Pereira AM, Roelfsema F, Smit JW, Hommes DW, Felt-BersmaRJ, RomijnJA.Acromegaly is associated with an increased prevalence of colonic diverticula: acase-control study. J Clin Endocrinol Metab.2010 May;95(5):2073-9.[br][br]Nothing to Disclose: EC, EH, MN, PK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 1073 76 823 SAT-745 PO22-01 Saturday 762 2012


763 ENDO12L_SAT-746 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Stomach and Duodenum Pathology at Patients with Acromegaly Alexander V Dreval, Sergey G Tereschenko, Anastasia Titaeva, Larisa E Gurevich, Nina A Korsakova, Irena A Ilovayskaya Moscow Regional Research [amp] Clinical Institute Named by MF Vladimirsky, Moscow, Russian Federation; Moscow Regional Research [amp] Clinical Institute Named by MF Vladimirsky, Moscow, Russian Federation; Moscow Regional Research [amp] Clinical Institute Named by MF Vladimirsky, Moscow, Russian Federation Stomach and duodenum conditions were estimated at 92 patients with active acromegaly (male 28, female 64, age 26-78 y.o., the majority of the patients 45-59 y.o.). Esophagogastroduodenoscopy was performed, including histological study in case of abnormality. Median GH levels were 22.7 [12.5; 51] mMe/l, median IGF-1 levels - 640 [507; 800] ng/ml. Esophagitis was diagnosed at 22 (23.9%) patients, incompetence of cardia - 33 (35.8%) patients. Gastroduodenitis was found in all patients (100%), additionally acute (n=4) of chronic (n=16) stomach erosions were revealed at 20 (21.7%) patients. Stomach polyps were found at 26 (27.3%) patients, polyps localized predominantly at distal parts of the stomach (mostly antrum), size up to 5 mm - 14 cases, 6-10 mm - 11 cases, [gt]10 mm - 3 cases. According to histological data, hyperplasia of surface epithelia was found in 16 (61.2%) cases, foveolar hyperplasia - in 4 (15.4%), and hyperplastic polyps - in 6 (23.4%) cases. Duodenal polyps were found at 2 other patients.[br]The majority of patients with acromegaly (78.5%) were contaminated with Helicobacter pylori: the 1st degree of contamination - 16.4%, the 2nd -16.4%, and the 3rd - 37.7% of patients. It was higher rate of contamination than in patients with gastroenterological diseases but without acromegaly (according to our experience - 48%). At 28 patients with stomach and duodenal polyps Helicobacter pylori was absent in 5 cases, the 1st degree of contamination was observed in 3, the 2nd - in 5, and the 3rd - in 15 cases. At 16 patients, who had chronic erosions of the stomach, the 1st degree of contamination was found at 3, the 2nd - at 1, and the 3rd - at 12 patients.[br]Thus, Helicobacter pylori contamination, stomach/duodenal polyps and different kinds of erosions are common among acromegalic patients. Not only colonic but also stomach and duodenal pathology are frequent complication of acromegaly. So, all patients with acromegaly need periodical complex observation of gastroenterology system.[br][br]Disclosures: AVD: Speaker, Lilly USA, LLC, Novo Nordisk, Novartis Pharmaceuticals, Ipsen. IAI: Speaker, Abbott Laboratories, Ipsen, Novartis Pharmaceuticals. Nothing to Disclose: SGT, AT, LEG, NAK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 505 76 824 SAT-746 PO22-01 Saturday 763 2012


764 ENDO12L_SAT-747 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Glucose Metabolism and Pituitary Pathology as Prognostic Factors in Acromegaly Sonia Cheng, Paula Araujo, Sylvia Asa, Shereen Ezzat University of Toronto, Toronto, Canada; University of Toronto, Toronto, Canada Introduction. Acromegaly is frequently associated with impaired glucose tolerance and/or diabetes. To evaluate the relationship between glucose metabolism and acromegaly disease prognosis we evaluated 127 consecutive patients.[br]Methods. We collected data on 127 acromegalic patients, including clinical features, pituitary tumor size, invasiveness on MRI, pituitary pathology, serum IGF-1, glucose at baseline and 120 minutes following an oral glucose tolerance test with corresponding growth hormone levels. IGF-1 at the time of the last visit was used to determine normalisation based on age and gender. Diabetes (DM) or impaired glucose tolerance (IGT) were defined according to 2011 ADA criteria.[br]Results. The study cohort included 63 women and 64 men with a mean age of 53.8 (SD 14.7). Of these 87 (68.5%) presented with a macroadenoma, 18 (14.2%) with a microadenoma, and 22 (17.3%) without definite radiological lesions. Radiographic invasiveness was present in 49 cases (38.6%), absent in 35 (27.6%), and could not be determined in 43 (33%). Mean tumor diameter was 1.86 cm (0.2-4.6). Pituitary histopathologic findings revealed pure somatotroph adenomas (SA) in 72 cases (56.7%), bihormonal lesions consisting of either mixed lactotroph adenoma (LA) with SA or mammosomatotroph adenoma (MSA) in 15 (11.8%), acidophil stem cell adenoma (ASA) in 2 (1.6%), and other diagnoses in 6 (4.7%). Trans-sphenoidal surgery (TSS) was performed in 117 (92.1%) cases, 95 (74.8%) were treated with adjunctive octreotide (OCT), 30 (23.6%) with pegvisomant (PEG), and 23 (18.1%) with dopamine agonists (DA); the latter two agents were used in combination with OCT. An equal proportion of patients with pure SA adenomas achieved IGF-1 normalization as those with mixed tumors (pure SA (76.7%) vs. 69.2%). Of note, however, DM or IGT were significantly more frequent (68.8% vs. 20%; p=0.009) in patients with pure SA adenomas and those with invasive tumors 66.7% vs. 41.7; p=0.06). Acromegalic patients with IGT/DM were also more likely (59.4% vs. 28.6%; p=0.054) to have persistently elevated IGF-1 levels independent of baseline GH, PRL, or IGF-1 levels.[br]Conclusions: IGT/DM at the time of diagnosis is more likely to be associated with pure SA adenomas and invasive tumors. Such patients are more likely to require adjunctive medical therapy to normalize IGF-1 levels.[br][br]Nothing to Disclose: SC, PA, SA, SE 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 321 76 825 SAT-747 PO22-01 Saturday 764 2012


765 ENDO12L_SAT-748 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Gender Differences in Thyroid Abnormalities at Patients with Acromegaly Tatiana P Shestakova, Alexander V Dreval, Olga A Nechaeva, Ekaterina A Zakharevich, Irena A Ilovayskaya Moscow Regional Research [amp] Clinical Institute Named by MF Vladimirsky, Moscow, Russian Federation There are gender differences in thyroid disorders at men and women. Thyroid abnormalities are frequent at patients with acromegaly. In purpose to elucidate gender differences in thyroid disorders at patients with acromegaly we examined 78 acromegalic patients: 12 male (M) [amp] 66 female (F), age 50.5[plusmn]17 [amp] 55.4[plusmn]12.6 y.o., duration of the acromegaly 4 [1.8; 6.3] [amp] 4.5[3.0; 6.0] y., GH levels 28.4 [12.2; 34.2] [amp] 9.1 [3.7; 25.3] mMe/l, IGF-1 724.5 [544.5; 964.3] [amp] 477 [246.3; 646] ng/ml, respectively. All patients underwent thyroid hormone testing, thyroid ultrasonografy, aspiration biopsy of thyroid nodules. Thyroid volume was 20 [13;25.6] ml M and 14 [9.75; 27.7] ml F. Normal thyroid structure was found in 4/12 (33.3%) M and 6/66 (9.1%) F (p=0.04). Diffuse goiter was revealed in 2/12 M and 11/66 F (both 16.7%). One solid nodule was observed in 3/12 (25%) M and 15/66 (22.7%) F. Multinodular goiter was found in 1/12 (8.3%) M and 30/66 (45.5%) F (p=0.02). Thyroid cancer (TC) was diagnosed in 2/12 (16.7%) M and 3/66 (4.5%) F (papillary cancer in all cases); men with TC had single thyroid nodule, and women with TC had multinodular goiter.[br]Hypothyroidism was observed in 3/12 (25%) M and 18/66 (27.3%) F: postsurgical hypothyroidism in 2 (16.7%) M (both after surgical treatment of TC) and 5 (7.6%) F (3 cases after surgical treatment of TC and 2 cases after surgical treatment of huge multinodular goiter); autoimmune thyroiditis in 1 (8.3%) M and 5 (7.6%) F; secondary hypothyroidism in 8 (12.1%) F only. There were also 2 cases of subclinical hyperthyroidism due to multinodular toxic goiter in women.[br]Thus, any thyroid structure abnormalities significantly more often can be found at women with acromegaly than in men. Multinodular goiter was predominantly observed in women whereas single thyroid nodule (often with papillary cancer) [ndash] in men. Hypothyroidism was found equally often at men and women with acromegaly, however, the main causes were different: postsurgical thyroid hypofunction in men and secondary hypothyroidism [ndash] in women.[br][br]Disclosures: AVD: Speaker, Lilly USA, LLC, Novo Nordisk, Novartis Pharmaceuticals, Ipsen. IAI: Speaker, Abbott Laboratories, Ipsen, Novartis Pharmaceuticals. Nothing to Disclose: TPS, OAN, EAZ 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 674 76 826 SAT-748 PO22-01 Saturday 765 2012


766 ENDO12L_SAT-749 POSTER SESSION: Pituitary Biology [amp] Acromegaly Co-Morbidities (1:30 PM-3:30 PM) Relationship between Nodular Thyroid Disease and Metabolic Paramethers in Patients with Acromegaly Tuncay Delibasi, Mustafa Sahin, Bekir Ucan, Nujen Colak Bozkurt, Erman Cakal, Mustafa Ozbek, Askin Gungunes, Basak Karbek, Evrim Cakir Ozkaya, Ilknur Ozturk Unsal, Muyesser Sayki Arslan, Esra Tutal Akkaymak, Taner Demirci, Melia Karakose Diskapi Yildirim Beyazit Training and Research Hospital, Ankara, Turkey; Ankara University School of Medicine, Ankara, Turkey The high prevalence of nodular goiter in patients with acromegaly is well known. Insulin-like growth factor-1 (IGF-1) has been claimed to be one of the etiologic factors. The aim of the study was to evaluate the incidence of thyroid lesions in our acromegalic patients and to analyse possible factors influencing thyroid nodule occurrence. Sixty patients with acromegaly, 32 females and 28 males, with a mean age of 52.7 [plusmn] 10. years without known thyroid disease in an iodine sufficient area were included. Age and sex matched 100 control subjects also included in the study. The frequency of nodular thyroid disease and metabolic parameters which may play a role in the formation of thyroid nodules were investigated. Waist and hip circumference, weight and height, fasting blood glucose, insulin, HbA1c, thyroid function tests, thyroid autoantibodies, lipid profile, vitamin B12, folic acid, CRP, vitamin D3, IGF-1, IGFBP-3, growth hormone, other anterior pituitary hormone levels were measured in all patients. Magnetic Resonance Imaging (MRI) and thyroid ultrasonography (US) were performed in all patients. Thyroid nodule volume, thyroid volume, HOMA-IR, HOMA-% B and HOMA% S were calculated.[br]Thirty-five (58.3%) patients with acromegaly had thyroid nodules according to (% 25) in control group (p[lt]0,0001). Also there were significant differences in BMI, thyroid volume, fasting glucose and TSH levels between patients and controls. After regression analysis, thyroid volume was associated with insulin, waist circumference, HOMA-IR, LDL-cholesterol and the size of the pituitary adenoma (p[lt]0.05) or diabetes occurence. During the logistic regression analysis, the presence of nodules was strongly associated with LH (p[lt]0.02) and HDL-cholesterol levels (p[lt]0.05). Nodule volume were significantly associated with luteinizing hormone (LH) level (p[lt]0.05), ACTH (beta = -0.51, p[lt] 0.01), plasma cortisol (beta= 0.965, p [lt]0.05), free T4, vitamin B12, folate, MCV, MPV, the size of adenoma in the regression analysis (p [lt]0.05) (Adjusted R square value = 0.882).[br]The prevalence of nodules in acromegalic patients (58%) were found to be higher than usual prevalence. Insulin resistance, vitamin D deficiency, growth hormone and IGF-1 levels have no effect on nodule prevalence in these patients. Hormones other than IGF-1 and insulin resistance might play an important role of thyroid volume, nodule volume and nodule formation mechanism in patients with acromegaly.[br][br]Nothing to Disclose: TD, MS, BU, NCB, EC, MO, AG, BK, ECO, IOU, MSA, ETA, TD, MK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 2113 76 827 SAT-749 PO22-01 Saturday 766 2012


767 ENDO12L_SAT-LB1 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Novel Primordial Dwarfism Syndrome Associated with Variants in the Centrosomal Protein Ninein Andrew Dauber, Stephen H LaFranchi, Zoltan Maliga, Julian C Lui, Jennifer E Moon, Cailin McDeed, Katrin Henke, Jonathan Zonana, Garrett A Kingman, Tune H Pers, Jeffrey Baron, Ron G Rosenfeld, Joel N Hirschhorn, Matthew P Harris, Vivian Hwa Children[apos]s Hospital Boston, Boston, MA; Broad Institute, Cambridge, MA; Oregon Health [amp] Science University, Portland, OR; Max Planck Institute for Cell Biology and Genetics, Dresden, Germany; Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD; Children[apos]s Hospital Boston, Boston, MA; Children[apos]s Hospital Boston, Boston, MA; Harvard University, Cambridge, MA Background: Microcephalic primordial dwarfism (MPD) is a rare, severe form of proportionate human growth failure in which growth restriction is evident in utero and continues into postnatal life. Single causative gene defects have been identified in a number of patients with MPD(1), and all involve genes fundamental to cellular processes such as genomic replication, DNA damage response, and centrosome functions.[br]Clinical Cases: We describe two sisters with severe IUGR who had progressive growth failure with final adult heights of 115 and 107 cm (-7.4 and -8.6 SDS) accompanied by profound microcephaly (head circumferences -7 SDS). Parental heights were normal. The probands presented with significant developmental delay and a seizure disorder. Evaluation of the growth hormone/IGF-1 axis was normal. Pubertal progression was absent and both sisters required estrogen supplementation. Physical features were not consistent with known syndromes of MPD.[br]Results: Whole exome sequencing of the two probands was undertaken and 34,606 shared variants were identified. All variants with minor allele frequency in databases greater than 1% were removed leaving 616 rare autosomal SNPs, from which 6 candidate genes were subsequently identified based on the presence of homozygous or [gt]1 heterozygous rare nonsynonymous variants. One of these genes was NIN, which encodes Ninein, a centrosomal protein involved in asymmetric mitotic spindle formation and appropriate development of radial glia progenitors (2,3). Our subjects were compound heterozygotes for two very rare missense variants in NIN (p.Q1222R/p.N1709S). Since dysfunctional centrosomes have been associated with MPD NIN was the most probable causal gene for MPD in our patients.[br]To explore the functional effects of NIN, we created a zebrafish model in which expression of ninein was reduced by morpholino methodology. Ninein knock-down led to specific defects in the specification and morphogenesis of the anterior neuroectoderm during early development of the larvae, resulting in patterning defects at the midbrain-hindbrain boundary. Additionally, reduction of Ninein resulted in deformity of the developing cranium with a small, squared skull highly reminiscent of the human phenotype.[br]Conclusion: We have identified a novel clinical subtype of MPD in two sisters who have rare variants in NIN. We show that reduction of ninein function during zebrafish development leads to specific deficiencies of brain and skull development.[br][br](1) Klingseisen A, Jackson AP 2011 Mechanisms and pathways of growth failure in primordial dwarfism. Genes Dev 25:2011-2024[br](2) Knoblich JA 2010 Asymmetric cell division: recent developments and their implications for tumour biology. Nat Rev Mol Cell Biol 11:849-860[br](3) Wang X, Tsai JW, Imai JH, Lian WN, Vallee RB, Shi SH 2009 Asymmetric centrosome inheritance maintains neural progenitors in the neocortex. Nature 461:947-955.[br][br]Sources of Research Support: NIH Grant 5K12HD052896 (AD); March of Dimes 6-FY09-507 (JNH), The Intramural Research Program of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health (JCL, JB).[br][br]Nothing to Disclose: AD, SHL, ZM, JCL, JEM, CM, KH, JZ, GAK, THP, JB, RGR, JNH, MPH, VH 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3034 77 828 SAT-LB1 PO01_LB Saturday 767 2012


768 ENDO12L_SAT-LB2 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Ultrastructural Remodeling of Pancreatic Islet Beta Cells in db/db Mice Melvin Hayden, Nivedita Nagam, James R Sowers University of Missouri, Columbia, MO; University of Missouri, Columbia, MO; University of Missouri, Columbia, MO; University of Missouri, Columbia, MO; Harry S Truman Memorial Veterans[apos] Hospital, Columbia, MO [bold]Introduction:[/bold] Obesity and type 2 diabetes mellitus (T2DM) are considered pandemic in adolescent and aging adult populations. We hypothesized that15 week old adolescent, obese, diabetic db/db mice models would demonstrate multiple beta ([beta])-cell ultrastructural remodeling as compared to their lean litter mate controls (C).[br][bold]Methods:[/bold] Thinly sliced tail specimens of the pancreas were placed immediately in standard transmission electron microscopy fixative; placed in resin and 85 nm thin sections were stained and viewed with a 1400 Joel electron microscope.[br][bold]Results:[/bold] Marked [beta]-cell hyperplasia and depletion of insulin secretory granules (ISG) were readily observed with increased surface area of endoplasmic reticulum perhaps reflecting ER stress in the db/db. Additionally, there was a loss of electron lucent immature insulin secretory bodies seen in concert with systemic insulin resistance and hyperinsulinemia. Noteable redistribution of [beta]-cell mitochondria with translocation to the perinuclear region of the [beta]-cells was observed. Importantly, cytoplasmic ectopic fat deposition consisting of excessive lipid droplets was noted in 15 week db/db mice as compared to the younger 12 week db/db mice and C mice.[br][bold]Discussion:[/bold] The marked remodeling in the db/db models may not only occur in adolescent, but also aged adult humans. These compensatory remodeling changes are associated with insulin resistance and hyperinsulinemia. Curiously, the15 week db/db models did not demonstrate any remodeling changes associated with apoptosis or autophagy.[br][bold]Conclusions:[/bold] Future studies utilizing life style changes and [beta]-cell sparing therapeutics in this model may prove to be beneficial these lipid deposition and [beta]-cell remodeling.[br][br]Sources of Research Support: National Institutes of Health R01 HL73101-08 and R01 HL107910-01; Veterans Affairs Merit System 0018 awarded to JRS.[br][br]Nothing to Disclose: MH, NN, JRS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3036 77 829 SAT-LB2 PO01_LB Saturday 768 2012


769 ENDO12L_SAT-LB3 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Effect of Peripheral Serotonin on Brown Adipose Tissue of Mice Fed High-Fat Diet Ryo Saito, Hitoshi Watanabe, Tatsuya Nakano, Keisuke Sumiyoshi, Yuya Nagasawa, Shunsuke Terada, Natsumi Okada, Kouichi Watanabe, Hisashi Aso Tohoku University, Sendai, Japan Brown adipose tissue (BAT) is known to be a thermogenesis organ because it has numbers of mitochondria including uncoupling protein 1 (UCP1) and burning off depot fat. Serotonin (5-hydroxytryptamine; 5HT) is a neurotransmitter in central nervous system and an intestinal hormone in peripheral tissue. Since serotonin is incapable of crossing the brain blood barrier, serotonin is thought to have independent function in central nervous system and periphery. On the function of central serotonin, a potentiation of 5HT within the intermediolateral cell column contributes to increases sympathetic nerve activity and thermogenesis in BAT, following the activation of 5HT receptors in the spinal cord. However, the functions of peripheral 5HT in BAT remain poorly understood. In this study, we investigated the role of peripheral 5HT in the activity of BAT.[br]Mice (C57BL/6, male) are fed chow diet or high-fat diet (HFD) from 5 weeks to 14 weeks of age. Mice in each diet were intraperitoneally injected with 5HT (1mg/mouse) or PBS twice a week. BAT was obtained from fasted mice at 14 weeks old. On the morphological characteristics of BAT, HE staining sections show that 5HT prevents HFD mice from the decrease of cell density in BAT. The transmission electron microscopy analysis revealed that 5HT increased the density of mitochondrial cristae in BAT of HFD mice. In addition, we investigated the number of mitochondria in BAT using the mitochondrial DNA / nuclear DNA quantification. The DNA copy numbers of mitochondrial component proteins in HFD mice were significantly increased by injection of 5HT. Moreover, 5HT markedly elevated the expression of UCP1 mRNA in BAT of HFD mice. The histological immunostaining revealed that the largest amount of UCP1 was also detected in BAT of HFD mice injected with 5HT. These data indicated that 5HT activated mitochondrial biosynthesis in BAT of HFD mice. Finally, BAT mitochondria were isolated mice in each diet and examined the mitochondrial capacity to oxidize fatty acid using a Clark type oxygen electrode. 5HT dramatically increased the mitochondrial oxidative capacity depended on proton leak of UCP1 in HFD mice. These findings show that the peripheral 5HT contributes to activate the mitochondrial biosynthesis and respiratory capacity of BAT in HFD mice.[br][br]Nothing to Disclose: RS, HW, TN, KS, YN, ST, NO, KW, HA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3056 77 830 SAT-LB3 PO01_LB Saturday 769 2012


770 ENDO12L_SAT-LB4 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Mdm2 and CREB-Binding Protein Regulates Estrogen Receptor [beta] Degradation in a PI3-K/Akt Dependent Manner Melanie Sanchez, Nathalie Picard, Karine Sauve, Andre Tremblay Ste-Justine Hospital, Montreal, Canada; University of Montreal, Montreal, Canada; University of Montreal, Montreal, Canada Estrogen receptors ER[alpha] and ER[beta] are regulated by estrogen and by growth factors signaling. Activation of ErbB2/ErbB3 tyrosine kinase receptors with growth factor heregulin leads to ER[beta] degradation, a mechanism that requires the coactivator cAMP response element-binding (CREB)-binding protein (CBP). CBP promotes ER[beta] polyubiquitination through enhancement of the PI3-K/Akt pathway by heregulin. Activated Akt triggered the recruitment of E3 ubiquitin ligase Mdm2 to ER[beta], which was further stabilized by CBP. Mutation of CBP Thr-1872 or Mdm2 Ser-186/188 Akt sites resulted in a dissociation of the ER[beta]-CBP-Mdm2 complex and reduced ER[beta] turnover. Treatment of breast cancer cells with heregulin led to a reduced occupancy of ER[beta] to target gene promoter and enhanced proliferation of breast cancer cells. However, knockdown of Mdm2 restored endogenous ER[beta] levels resulting in reduction of breast cancer cell growth. These studies identify a tripartite Akt-regulated phosphorylation mechanism that functions to hamper normal ER[beta] activity and turnover through the concerted actions of CBP and Mdm2 in response to growth factor signaling pathways in breast cancer cells.[br][br]Nothing to Disclose: MS, NP, KS, AT 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3061 77 831 SAT-LB4 PO01_LB Saturday 770 2012


771 ENDO12L_SAT-LB5 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Adipose-Specific Deletion of Protein-Tyrosine Phosphatase 1B Alters Energy Balance in Mice Ahmed Bettaieb, Fawaz George Haj, Kosuke Matsuo, Izumi Matsuo University of California-Davis, Davis, CA; Yokohama City University, Yokohama, Japan Objective: Protein-tyrosine phosphatase 1B is an important physiological regulator of adiposity and glucose homeostasis. Mice with whole-body PTP1B deletion exhibit increased insulin sensitivity and are protected from high fat diet-induced obesity. However, the main sites and mechanisms of PTP1B action have not been fully elucidated. The objective of this study is to determine the metabolic role of adipose PTP1B. Research Design and Methods: To address the potential role of adipose PTP1B in regulating adiposity and insulin sensitivity, we generated mice with adipose-specific PTP1B deletion using the novel Adiponectin-Cre transgenic mice. The effects of efficient and adipose-specific PTP1B deletion on adiposity and glucose homeostasis were assessed. Results: We report that adipose-specific PTP1B deletion reduces body weight and adiposity in male and female mice on high fat diet. This is a result, at least in part, of increased energy expenditure in these mice. In addition, adipose PTP1B deletion improves systemic insulin sensitivity and glucose homeostasis. These findings correlate with enhanced tyrosyl phosphorylation of the insulin receptor and its downstream signaling pathway in KO mice. Conclusions: These studies reveal that Adiponectin-Cre mice are a useful tool to achieve efficient and adipose-specific deletion. In addition, our findings indicate that targeting PTP1B in adipose tissue may be a useful approach for treatment and prevention of obesity and insulin resistance.[br][br]Sources of Research Support: Junior Faculty Award from the American Diabetes Association.[br][br]Nothing to Disclose: AB, FGH, KM, IM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3084 77 832 SAT-LB5 PO01_LB Saturday 771 2012


772 ENDO12L_SAT-LB6 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) CAPER Is an Essential Coordinator for Transcriptional Regulation of Cellular Bioenergetics Yun Kyoung Kang, Olga R Ilkeyava, Subhamoy Dasgupta, Sophia Y Tsai, Ming-Jer Tsai, Christopher B Newgard, Bert W O[apos]Malley Baylor College of Medicine, Houston, TX; Duke University Medical School, Durham, NC Maintenance of energy homeostasis is the crucial prerequisite to sustain life. Here we present roles for CAPER as an essential coactivator to orchestrate multitudes of metabolic processes for ATP homeostasis. Depriving cells of CAPER induces characteristic starvation responses [ndash]decreased cell number, autophagic vacuolization and increased phospho-AMPK and eventually depletion of ATP. Depleting CAPER results in lower mitochondrial respiratory capacity and fewer transcripts of mitochondrial genes without much change in total numbers of mitochondria. Metabolomic profiling and metabolite flux assays demonstrated that the knock down of CAPER reduced the anaplerotic influx to the TCA cycle, including glycolysis, amino acid catabolism and fatty acid oxidation, as a result of decreased expressions of the corresponding enzymes. We found that CAPER works with multiple transcription factors including c-Myc and ERR alpha to regulate its target genes. Taken together, our studies present a regulatory mechanism by which cells maintain ATP homeostasis through CAPER by achieving simultaneous control of multiple transcriptional nodes in energy metabolism.[br][br]Sources of Research Support: HD 08818.[br][br]Nothing to Disclose: YKK, ORI, SD, SYT, M-JT, CBN, BWO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3005 77 833 SAT-LB6 PO01_LB Saturday 772 2012


773 ENDO12L_SAT-LB7 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) In Vitro Study of the Role of Histone Modifications in Steroid Receptor-Mediated Transcription Qin Feng, Charles E Foulds, Suzanna L Bailey, Ross A Hamilton, Anna Malovannaya, David M Lonard, Jun Qin, Bert W O[apos]Malley Baylor College of Medicine, Houston, TX As fundamental structural units of chromatin, core histones are subject to multiple post-translational modifications such as acetylation, phosphorylation, methylation, ubiquitination, ADP-ribosylation, and recently identified, crotonylation. The precise function of each histone modification remains largely unknown. We hypothesize that differentially modified histones have profound effects on the formation of steroid receptor and coregulator complex on the target gene promoter and the subsequent transcription activation.[br]Based on an adenoviral E4 gene reporter DNA which harbors several copies of steroid receptor DNA binding elements (SREs), we have established a transcription system using in vitro assembled nucleosomal E4 DNA as a template. To assemble the nucleosomes, in addition to the traditionally used core histone octamers that are purified from HeLa cells and contain poorly defined post-translational modifications, we also use recombinant histones with precisely defined modification. We believe, using this system as a tool, we might be able to dissect the functional interactions between histone modifications, receptor/coregulator assembly, and transcription activation. For instance, we have demonstrated that tri-methylation of histone H3 at K4 is indeed an essential step to ensure the optimal transcription activation. The functions of other histone modifications are being tested in our system.[br]By unbiased mass spectrum analysis, the transcriptional coregulator complexes bound to ER[alpha] on nucleosomal E4 DNA with H3K4me3 modification were identified. Compared with the coregulator complex associated with nucleosomes containing H3K9me3 modification, we found H3K4me3 modification dramatically enhances ER and coregulator recruitment in nucleosome-based IVT.[br]Our result indicates that the nucleosome-based IVT system is a novel and excellent tool to dissect the role of histone modifications in steroid receptor-mediated transcription.[br][br]Sources of Research Support: Partially by NIH K01DK084209 awarded to Qin Feng.[br][br]Nothing to Disclose: QF, CEF, SLB, RAH, AM, DML, JQ, BWO 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3013 77 834 SAT-LB7 PO01_LB Saturday 773 2012


774 ENDO12L_SAT-LB8 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) The Role of ATG12 in Hypothalamic Function Ritu Malhotra, James P Warne, Eduardo Salas, Allison W Xu, Jayanta Debnath UCSF, San Francisco, CA; UCSF, San Francisco, CA Autophagy is a fundamental lysosomal degradation process that requires multiple proteins called ATGs. Although the prevailing view has been that ATGs are exclusively devoted to autophagy, there is increasing recognition that individual ATGs also have unique functions.[br]To test this hypothesis in vivo, we created complete and conditional Atg12 deletion mice. Similar to Atg5 deficiency, complete Atg12 null mice die on postnatal day 1, corroborating an essential role for ATG12-ATG5 for autophagy during the neonatal period. Notably, Atg12 (but not Atg5) transcripts are highly enriched in the hypothalamus, which play critical role in mammalian food intake and energy balance. This finding motivated us to delete Atg12 in hypothalamic POMC neurons. Although recent studies have shown deletion of another autophagy regulator, Atg7, in POMC neurons leads to increased body weight due to hyperphagia, we have found that Atg12 deletion, has no effect on body weight or adiposity in these mice. Furthermore, Atg12F/F-Pomc Cre mice have similar feeding and energy expenditure as wild type controls. Remarkably, males lacking Atg12 in POMC neurons exhibit improved glucose tolerance, although they exhibit no difference in their insulin sensitivity.[br]Based on these findings, we dissected how the loss ATG12 in POMC neurons regulated diet-induced obesity. Mice fed a high-fat diet (HFD) display increased induction of autophagosomes in POMC neurons. Furthermore, Atg12F/F-Pomc Cre mice on HFD show a dramatic increase in body weight, which is accompanied with impaired glucose tolerance. These mice show increased food intake and reduced ambulatory activity, leading to reduced energy expenditure. The mRNA expression of POMC is significantly reduced in Atg12F/F-Pomc Cre mice on HFD. We also created POMC Atg5 deficient mice to parse if this phenotype is due to a general defect in autophagy versus a unique function mediated by ATG12. Similar to Atg12F/F-Pomc Cre mice, Atg5F/F-Pomc Cre mice do not show any differences in body weight or in adiposity. Overall, these data do point to a general role for autophagy in the modulation of POMC neuronal function in response to high fat diet; nonetheless, in the absence of this stressor, the loss of individual ATGs each elicits unique effects on basal hypothalamic function. These studies also illustrate importance of genetically interrogating multiple ATGs prior to ascribing a general role for autophagy in mediating physiological functions in vivo.[br][br]Nothing to Disclose: RM, JPW, ES, AWX, JD 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3006 77 835 SAT-LB8 PO01_LB Saturday 774 2012


775 ENDO12L_SAT-LB9 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) A Noncompetitive Small Molecule Antagonist of Progesterone Receptor That Inhibits Nuclear Localization and Gene Expression Irene O Aninye, Steven K Nordeen, David J Shapiro University of Illinois at Urbana-Champaign, Urbana, IL; University of Colorado Anschutz Medical Campus, Aurora, CO; University of Illinois at Urbana-Champaign, Urbana, IL Small molecule inhibitors have the potential to be powerful probes of steroid receptor action. However, only a few noncompetitive antagonists of progesterone receptor (PR) have been described. Screening for PR inhibitors led to the identification of a novel noncompetitive small molecule inhibitor of PR-mediated gene expression that acts, at least in part, by inhibiting nuclear localization of liganded PR. PR plays a key role in reproduction and is important in cancers of the reproductive tract. Current PR antagonists usually act by competing with progestins for binding in the receptor[apos]s ligand-binding pocket and often exhibit cross-reactivity with other members of the steroid receptor family. Using a cell-based screen, we identified alkylthio-substituted theophyllines that possess a novel structural motif and selectively inhibit PR-mediated gene expression. These are noncompetitive inhibitors acting outside the ligand-binding pocket of the receptor. The lead compound, PR51, did not induce degradation of PR, nor did it inhibit binding of PR to a consensus PRE in an [italic]in vitro[/italic] fluorescence anisotropy assay. However, chromatin immunoprecipitation in T47D cells showed that treatment with PR51 greatly reduced recruitment of PR to the enhancer region of [italic]sgk1[/italic]. When nuclear and cytoplasmic extracts were isolated and probed for PR content, PR51 promoted cytoplasmic retention of progestin-PR and inhibited nuclear localization of ligand-bound receptor. PR51 and its relatives are a novel class of PR antagonists that disrupt the nuclear localization of liganded PR. Thus, PR51 represents a powerful new probe for dissecting PR action.[br][br]Sources of Research Support: NIH Grant RO1DK0719019; NIH Ruth L. Kirschstein NRSA F31DK0848545.[br][br]Nothing to Disclose: IOA, SKN, DJS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3014 77 836 SAT-LB9 PO01_LB Saturday 775 2012


776 ENDO12L_SAT-LB10 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Melanocortin 4 Receptors Contribute to Stress-Induced Activation of the HPA Axis Karen K Ryan, Joram D Mul, Denovan P Begg, Christoffer Clemmensen, Kristen N Halcomb, Randy J Seeley, James P Herman, Yvonne M Ulrich-Lai University of Cincinnati, Cincinnati, OH; University of Cincinnati, Cincinnati, OH The melanocortin 4 receptor (MC4R) is widely expressed in stress-regulatory brain regions, including the paraventricular nucleus of the hypothalamus, and has been implicated in HPA regulation. The present work determined whether MC4R signaling is necessary for the HPA axis response to stress using a recently characterized rat model with a loss-of-function mutation in [italic]Mc4r[/italic]. This mutation prevents receptor membrane binding and signaling. Rats were bred on site, and adult males were placed into 3 groups (n=12/group) based on genotype (wild type (WT); heterozygous mutant (HET); and homozygous mutant (HOM)). Basal (pre-stress) plasma ACTH and corticosterone were measured in the AM and PM, and the HPA axis response to restraint was assessed in the AM. Rats were perfused at 2 hours after restraint to assess the effects of loss of MC4R on stress-induced c-fos immunolabeling in stress-regulatory brain regions. Relative to WT, HOM rats had increased body weight, whereas HET rats showed an intermediate phenotype. Basal (non-stress) AM and PM plasma corticosterone and ACTH showed a normal diurnal rhythm that was not altered in either HET or HOM rats. In addition, adrenal and thymus weights were unaffected by genotype. However, the plasma ACTH and corticosterone responses to restraint were reduced in HOM rats, and this attenuation was less pronounced in HET rats. In summary, loss of MC4R function results in diminished HPA axis stress responsivity, without affecting basal HPA axis tone. These results support the hypothesis that endogenous MC4R signaling contributes to the HPA axis response to stress. Given that MC4R signaling plays a critical role in energy balance regulation, the present work suggests that it may also serve as an important communication link between brain metabolic and stress systems.[br][br]Sources of Research Support: NIH grants R01 MH049698 (JPH), K01 DK078906 (YMU), R03 DK089018 (YMU), and R01 DK091425 (YMU).[br][br]Nothing to Disclose: KKR, JDM, DPB, CC, KNH, RJS, JPH, YMU-L 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3024 77 837 SAT-LB10 PO01_LB Saturday 776 2012


777 ENDO12L_SAT-LB11 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Membrane-Initiated Effects of Progestogens Alone and in Combination with Estradiol on the Proliferation of Human Breast Cancer Cells Hans Neubauer, Xiangyan Ruan, Helen Schneck, Harald Seeger, Michael A Cahill, Tanja Fehm, Yayun Liang, Benford Mafuvadze, Hyder M Salman, Alfred O Mueck Eberhard Karls University, Tuebingen, Germany; Beijing Ob/Gyn Hospital, Capital Medical University, Beijing, China; Charles Sturt University, Wagga Wagga, Australia; University of Missouri, Columbia, MO Objectives: Progestogens may play a crucial role in the development of breast cancer under contraception and hormone therapy. Progesterone receptor membrane component-1 (PGRMC1) may be important in tumorigenesis and thus may increase breast cancer risk. We investigated PGRMC1 expression in human breast tissues and the influence of progestogens alone and in combination with estradiol on breast cancer cells overexpressing PGRMC1 and tumor growth in a mouse transplantation model.[br]Methods: PGMRC1 expression was measured immunhistochemically in 69 breast cancer tissues. Progesterone (P), chlormadinone acetate (CMA), desogestrel, drospirenone, dydrogesterone, levonorgestrel, medroxyprogesterone acetate, nomegestrol (NOM) and norethisterone (NET) were tested alone or sequentially and continuously combined with two estradiol concentrations in MCF-7 cells transfected with PGRMC1 (WT-12 cells). Cell proliferation was measured by MTT assay. In a transplantation model six weeks old nude mice were inoculated with E2 pellets (0.72mg/60 day release) 24 hours before tumor cell injections into both flanks. After eight days animals were inoculated with a NET pellet (10 mg/60 day release) or control pellets, and tumor volumes were recorded twice a week. Empty vector transfected cells served as controls in all proliferation experiments.[br]Results: PGRMC1 expression is significantly higher in cancerous tissues than in benign tissues. In vitro the greatest stimulating effect on WT-12 cells was found for NET. No proliferative increase was observed for CMA, NOM and P. Estradiol/progestogen combinations elicited an estradiol-concentration dependent effect. At high estradiol levels the estradiol-induced proliferative effect dominates. At low estradiol concentrations the effect of the synthetic progestins is prevailing. The proliferative gain was lower with sequential combination than with continuous combination. In vivo NET sequentially combined with E2 also significantly increased tumor growth of WT-12 cells.[br]Conclusion: Some synthetic progestins trigger a proliferative response of PGRMC1 overexpressing MCF-7 cancer cells. The effect of progestogens on breast cancer tumorigenesis may clearly depend on the specific pharmacology of the various synthetic progestins. In combination with estradiol the use of specific progestogens, the kind of regimen as well as the use of low estradiol dosages might be of importance when applying hormone therapy for contraception and in the postmenopause.[br][br]Nothing to Disclose: HN, XR, HS, HS, MAC, TF, YL, BM, HMS, AOM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3027 77 838 SAT-LB11 PO01_LB Saturday 777 2012


778 ENDO12L_SAT-LB12 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Differential Regulation of Beta-Cell Insulin Secretion by Ghrelin and Somatostatin Is Dependent upon Energy Balance and Equilibrium between GHSR1a Homodimers and GHSR1a:SST5 Heteromers Seongjoon Park, Hong Jiang, Roy G Smith The Scripps Research Institute, Florida, Jupiter, FL; Nagasaki University (current institution), Japan City, Japan; University of Texas Health Science Center, Houston, TX The gut-derived hormone ghrelin regulates growth hormone release, appetite and glucose homeostasis. Ghrelin activation of its receptor (GHSR1a) normally results in coupling to Ga[sub]q11[/sub] producing an increase in [Ca[sup]2+[/sup]][sub]I[/sub] which should enhance glucose-stimulated insulin secretion (GSIS) by pancreatic beta-cells; instead, ghrelin inhibits GSIS by a mechanism involving coupling of GHSR1a to Ga[sub]i2[/sub] (Dezaki et al. 2007). Besides ghrelin, somatostatin (SST) inhibits GSIS. How are ghrelin and SST actions independently regulated and under what physiological conditions? The mechanism must accommodate non-canonical ghrelin receptor (GHS-R1a) G-protein coupling and dependence on energy balance. We quantitated expression of GHSR1a, SST5, SST2, ghrelin and GOAT in rat islets and developed a system based on a new subclone of the INS-1 beta-cell line (INS-1SJ) to test our hypothesis that ghrelin inhibition of GSIS via Ga[sub]i[/sub] signaling is dependent upon interactions between equimolar concentrations of GHSR1a and SST5. The INS-1SJ cells express SST2, ghrelin and GOAT and we defined conditions of electrophoration to recapitulate relative and absolute expression levels of GHSR1a and SST5 in islets. Low glucose (3 mM) increases ghrelin and treating these cells with exogenous ghrelin does not inhibit GSIS. Treating these cells with ghrelin-siRNA enhances GSIS and cAMP accumulation which is reversed by exogenous ghrelin treatment, thereby supporting a role for inhibition of GSIS by endogenous ghrelin. Intriguingly, although ghrelin inhibits GSIS, the SST5 agonist BIM23052 is ineffective in the presence of GHSR1a. Tr-FRET and BRET analyses show that GHSR1a non-canonical G-protein coupling via Ga[sub]i[/sub] is dependent on formation of GHSR1a:SST5 heteromers. In contrast to SST5, SST2 does not form heteromers with GHSR1a and is not required for ghrelin inhibition of GSIS. BRET analysis is consistent with stabilization of GHSR1a:SST5 heteromers and Ga[sub]i/o[/sub] coupling by ghrelin, whereas SST appears to destabilize heteromer formation.[br][italic]In vivo [/italic]when glucose is low, [ghrelin] is high and [SST] is low. When glucose is high the reverse is true. Our collective data are consistent with an equilibrium model of receptor heteromerization dependent upon the ratio of [ghrelin]/[SST] that is controlled by glucose concentrations. This mechanism allows differential regulation of pancreatic b-cell function by ghrelin and SST according to energy balance.[br][br]Sources of Research Support: The support of R01AG19230 and R01AG29740 to Roy G. Smith is gratefully acknowledged.[br][br]Nothing to Disclose: SP, HJ, RGS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3031 77 839 SAT-LB12 PO01_LB Saturday 778 2012


779 ENDO12L_SAT-LB13 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Gene Engineered T-Cells for the Immunotherapy of Differentiated Thyroid Cancer Brandon J Baird, Brock J Lanier, Ken-ichi Hanada, Qiong J Wang, Zhiya Yu, Youn Kim, Nicholas P Restifo, James C Yang National Cancer Institute, National Institutes of Health, Bethesda, MD; Duke University, School of Medicine, Durham, NC The incidence of thyroid cancer is rapidly increasing in the United States. Though mortality for differentiated thyroid cancer is low with a 10-year survival rate [gt]90%, it increases dramatically with metastatic disease for which there are few therapeutic options. The transfer of cultured tumor-reactive T cells or adoptive cellular therapy (ACT) has demonstrated high rates of objective tumor regression in patients with refractory melanoma, lymphoma and synovial cell sarcoma. Some patients with melanoma have achieved complete tumor regressions, which are ongoing 5-8 years after cell transfer. Therapeutic T-cells for transfer can either be obtained from tumor-infiltrating lymphocytes in melanoma or genetically engineered from peripheral blood lymphocytes (PBL). A major obstacle in the treatment of other cancers using ACT has been the paucity of good target antigens. Thyroglobulin (TG) is an ideal target because of its high level of expression limited to differentiated thyroid cancer and normal thyroid, a dispensable tissue. Using a mouse transgenic for HLA-A*0201 (an allele with 50% prevalence in the U.S.) and Yeti, a reporter system that drives the expression of yellow fluorescent protein (YFP) by the interferon-gamma (IFN-g) promoter, we generated HLA-A*0201-restricted murine T cells specific for the aa 470-478 epitope of TG by vaccinating with an adenovirus that encodes human TG. Previous clinical and laboratory studies have demonstrated that murine T-cell receptors (TCR) engage human CD3 and function when introduced into human lymphocytes. TCRs from these TG-reactive murine T-cells were cloned into a retroviral vector, which was able to transduce human peripheral blood lymphocytes (PBL) with 80-90% efficiency. These transduced PBL showed high levels of human IFN-g release in response to primary thyroid tissue, a thyroid tumor line, and cells transfected with TG in the context of HLA-A*0201, while showing no reactivity to non-cognate antigens. A clinical protocol transferring autologous TCR-engineered PBL into HLA-A*0201+ patients with metastatic differentiated thyroid cancer is in development.[br][br]Sources of Research Support: This research year was made possible through the Clinical Research Training Program, a public-private partnership supported jointly by the NIH and Pfizer Inc (via a grant to the Foundation for NIH from Pfizer Inc).[br][br]BJB: Recipient Award, Pfizer, Inc. Nothing to Disclose: , BJL, K-IH, QJW, ZY, YK, NPR, JCY 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3033 77 840 SAT-LB13 PO01_LB Saturday 779 2012


780 ENDO12L_SAT-LB14 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Transcriptome of Isolated Endometrial Cell Populations in Women with PCOS Reveals Increased Immune Cell Trafficking, Cell Growth and Predisposition to Endometrial Cancer Terhi T Piltonen, Trimble LB Spitzer, Fatima Barragan, Joseph Chen, Juan C Irwin, Heather Huddleston, Linda C Giudice University of California, San Francisco, CA Polycystic ovary syndrome (PCOS) is generally associated with anovulatory sub-fertility and increased risks of miscarriage and endometrial cancer. Progesterone resistance and a proliferative phenotype in endometrium likely contribute to these risks, with obesity, common in PCOS (50-70%), aggravating symptoms and endometrial dysfunction. To address our hypothesis that an altered metabolic/ endocrine/immune environment may result in a [ldquo]diseased niche[rdquo] for endometrial stem cells and their differentiated progeny, we determined the transcriptome and biological processes/pathways of isolated cell populations, including endometrial mesenchymal stem cells (eMSC) (progenitor of the stromal fibroblast (eSF)), in endometrium of overweight/obese PCOS women compared to overweight/obese controls.[br][bold]Materials and Methods:[/bold] Proliferative phase endometrial tissue samples were obtained from PCOS (n=5, NIH criteria, age 31.2[plusmn]5.5 yrs, BMI 34.23[plusmn]6.1) and control women (n=4, age 35.75[plusmn]5.0 BMI 38.93[plusmn]10.8). Microarray (Affymetrix Human Gene 1.0 ST array), bioinformatic and Ingenuity Pathway (IPA) analyses were performed on FACS-sorted eSF (CD146-/PDGFRB+), epithelium (EP, EpCam+/CD45-), endothelium (EN, CD146+/PDGFRB-) and eMSC (CD146+/PDGFRB+).[br][bold]Results:[/bold] Samples, analyzed by principal component and hierarchical clustering analyses, clustered by cell type and disease or control. In eSF[sub]pcos[/sub] vs. eSF[sub]ctrl[/sub] 427 genes were differentially expressed, with up-regulation of genes related to immune cell trafficking ([italic]CCl2, ICAM1, TNFAIP3[/italic]). Of 993 genes differentially expressed in EP[sub]pcos[/sub] vs. EP[sub]ctrl[/sub] was up-regulation of inflammatory genes ([italic]ORM1/2, IL6, CCL2[/italic]); the most down-regulated genes were [italic]SFRP4[/italic] and [italic]SPARC[/italic] (decreased in endometrial adenocarcinoma). Subtle differences were observed in the endothelial populations; whereas, among the 324 differentially expressed genes between eMSC[sub]pcos[/sub] and eMSC[sub]ctrl[/sub] the most up-regulated genes were [italic]LCN2[/italic] (a growth factor), [italic]IL8[/italic] and [italic]CXCL2[/italic]. According to IPA the top biological functions included cancer, inflammation, immune response, cell growth and proliferation.[br][bold]Conclusions:[/bold] Global gene expression analysis of isolated endometrial cell populations in overweight/obese women with PCOS vs. overweight/obese controls showed increased inflammation and cell growth. Cancer-related changes were observed especially in epithelium. That the transcriptome of eMSC is altered in PCOS supports the hypothesis of a [ldquo]diseased niche[rdquo] with potential effects on lineage differentiated cells.[br][br]Sources of Research Support: Grant support: Sigrid Juselius Foundation, Finnish Academy, Orion-Farmos Research Foundation, Maud Kuistila Foundation (TTP) and NIH U54HD 055764-05 Specialized Cooperative Centers Program in Reproduction and Infertility Research (LCG).[br][br]Nothing to Disclose: TTP, TLBS, FB, JC, JCI, HH, LCG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3037 77 841 SAT-LB14 PO01_LB Saturday 780 2012


781 ENDO12L_SAT-LB15 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Protein Arginine Methyltransferase 6 Dependent Gene Expression and Splicing: Association with Breast Cancer Outcomes Dennis H Dowhan, Matthew J Harrison, Natalie A Eriksson, Peter Bailey, Michael A Pearen, Peter J Fuller, John W Funder, Evan R Simpson, Peter J Leedman, Wayne D Tilley, Melissa A Brown, Christine L Clarke, George EO Muscat The University of Queensland, Brisbane, Queensland, Australia; The University of Queensland, Brisbane, Queensland, Australia; Prince Henry[apos]s Institute for Medical Research, Clayton, Victoria, Australia; Western Australian Institute for Medical Research and University of Western Australia Centre for Medical Research, Perth, Western Australia, Australia; The University of Adelaide, Hanson Institute, Adelaide, South Australia, Australia; University of Sydney, Western Clinical School, Westmead, New South Wales, Australia Protein arginine methyltransferases (PRMTs) catalyze post-translational modifications that regulate transcription and RNA processing. We have recently demonstrated that PRMT6 effects steroid hormone mediated transcription and inclusion:skipping ratio of alternatively spliced exons in vascular endothelial growth factor and the tumour suppressor spleen tyrosine kinase genes. In this study we examined the role PRMT6 has in regulating gene expression in breast cancer cells and the corresponding relationship between a PRMT6 gene signature and breast cancer patient clinical outcomes. To identify novel gene targets regulated by PRMT6 in breast cancer cells, we used a combination of siRNA, exon-specific microarray profiling in MCF-7 breast cancer cells in vitro. Verification of PRMT6 gene expression targets was examined in breast cancer cell lines, human normal breast tissue and primary human breast tumours by quantitative real-time PCR. We then evaluated the PRMT6 transcriptionally regulated gene signature in the context of clinical outcome associations in three publically available independent breast cancer datasets. This approach, which allows the examination of genome-wide changes in individual exon usage and total transcript levels, demonstrated PRMT6 knockdown significantly affected: (i) the transcription of 159 genes, and (ii) alternative splicing of 449 genes. Importantly, the levels of PRMT6 itself were significantly decreased in breast cancer, relative to normal breast tissue. The PRMT6 dependent transcriptional and alternative splicing targets identified in vitro, were validated in human breast tumours. Notably, we found that reduced PRMT6 expression and the corresponding gene expression signature, correlated with decreased probability of relapse free or metastasis free survival in estrogen receptor (ER)+ breast cancer. PRMT6 can determine transcriptional gene regulation and alternative splicing decisions, making it a master-modulator of gene expression. A significant number of genes regulated by PRMT6 are over-represented by genes involved in important aspects of carcinogenesis and a PRMT6 dependent gene signature is significantly associated with clinical outcome in ER+ breast cancer. These results suggest that dysregulation of PRMT6 dependent transcription and alternative splicing may be involved in breast cancer pathophysiology and the molecular consequences identifying a unique and informative biomarker profile.[br][br]Sources of Research Support: This work was supported by a National Health and Medical Research Council of Australia (NHMRC) project grant (Grant#511153) to DHD. MJH was the recipient of an ANZ Trustees PhD Medical Research scholarship. In addition, this research was supported by a grant from the National Breast Cancer Foundation (NBCF) Australia.[br][br]Nothing to Disclose: DHD, MJH, NAE, PB, MAP, PJF, JWF, ERS, PJL, WDT, MAB, CLC, GEOM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3049 77 842 SAT-LB15 PO01_LB Saturday 781 2012


782 ENDO12L_SAT-LB16 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Genetic Variation in Susceptibility to Hypothyroidism Induced Hearing Impairment Thomas J Jones, Qing Fang, Tzy-Wen Gong, Amanda H Mortensen, Michelle T Fleming, David Dolan, Margaret Lomax, Kenneth R Johnson, Mirna Mustapha, Sally A Camper University of Michigan Medical School, Ann Arbor, MI; University of Michigan Medical School, Ann Arbor, MI; Stanford University, Stanford, CA; Jackson Laboratory, Bar Harbor, ME Hypothyroidism is a common health problem, and if present in pregnant mothers or newborns it may cause hearing impairment and cognitive disability in the child. Individuals vary in their susceptibility to these associated problems. To understand why patients differ in response to hypothyroidism, we used congenitally hypothyroid mouse mutants because they offer the advantages of tissue accessibility and unique genetic backgrounds. We discovered that the genetic background of the fetus confers susceptibility to or protection from hearing impairment (1, 2). For example, mutations in [italic]Pou1f1[/italic] cause severe secondary hypothyroidism and dwarfism regardless of the genetic background, but the mutants are profoundly deaf on the susceptible DW/J and C57BL/6J strains and have only mild hearing impairment on the protective C3H/HeJ strain (3). Hypothyroidism has pleiotropic effects on middle ear and cochlear development in profoundly deaf mutants. Hypothyroid mutants have developmental delay that affects organ of Corti morphology, sensory cell innervation, and expression of genes important for normal potassium flux (1,2,4). Here we report genetic mapping of a locus in both CAST/Ei and 129P2/OlaHsd strains that confers protection against hearing impairment in DW/J-[italic]Pou1f1[/italic][sup]dw/dw[/sup] mutants. 377 F2 mutant animals from F1 intercrosses were assessed for hearing ability by auditory brain stem response and genotyped for 1500 SNP markers. We identified a single locus on chromosome 2 that we named modifier of [italic]dw[/italic] hearing, [italic]Mdwh[/italic]. The 20 Mb [italic]Mdwh[/italic] critical region contains over 500 genes, and DNA sequence comparisons reveal few variants predicted to alter gene function. To narrow the field of candidate genes we compared the cochlear gene expression profiles of hypothyroid DW/J-[italic]Pou1f1[/italic] mutants and wild type mice. Over 500 thyroid hormone regulated genes were identified, and ten mapped to the [italic]Mdwh[/italic] region. We are comparing expression levels of these 10 genes in susceptible and protective strains to determine which is responsible for the protective effect. Our catalog of genes regulated by thyroid hormone in the cochlea is a rich resource for understanding the mechanism of hypothyroidism induced hearing impairment, and identification of the protective [italic]Mdwh[/italic] gene could be a foundation for developing new therapies for human patients that are not responsive to thyroid hormone replacement.[br][br](1) Karolyi IJ et al., Mamm Genome 18:596-608, 2007.[br](2) Fang Q et al., J Assoc Res Otolaryngol 13:173-184, 2012.[br](3) Fang Q et al., Genetics 189:665-673, 2011.[br](4) Mustapha M et al., J Neurosci 29:1212-1223, 2009.[br][br]Sources of Research Support: Endocrine Society Summer Fellowship (TJJ); National Institutes of Health (R01DC009590 MM) and P30 DC05188 (DFD); March of Dimes #6-FY08-262 (SAC).[br][br]Nothing to Disclose: TJJ, QF, T-WG, AHM, MTF, DD, ML, KRJ, MM, SAC 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3074 77 843 SAT-LB16 PO01_LB Saturday 782 2012


783 ENDO12L_SAT-LB17 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Requirement for Stromal Estrogen Receptor Alpha in Cervical Neoplasia Sang-Hyuk Chung, Myeong-Kyun Shin, Kenneth S Korach, Paul F Lambert University of Wisconsin, Madison, WI; University of Houston, Houston, TX; National Institute of Environmental Health Sciences, Research Triangle Park, NC The major etiological factor for cervical cancer is the high-risk human papillomavirus (HPV), which encodes E6 and E7 oncogenes. However, HPV is not sufficient and estrogen has been proposed as an etiological cofactor for the disease. Its requirement has been demonstrated in mouse models for HPV-associated cervical cancer (e.g., K14E7 transgenic mice). Although germline knockout of estrogen receptor alpha (ER[alpha]) renders mice resistant to cervical cancer, the cell type-specific requirement for ER[alpha] is not known. We demonstrate that temporal deletion of stromal ER[alpha] induced complete regression of cervical dysplasia in K14E7 mice. Our results strongly support the hypothesis that stromal ER[alpha] is necessary for HPV-induced cervical carcinogenesis. Our results implicate paracrine mechanisms involving ER[alpha] signaling contribute to development of estrogen-dependent cervical cancers. This is the first evidence to support the importance of stromal ER[alpha] in estrogen-dependent neoplastic disease.[br][br]Nothing to Disclose: S-HC, M-KS, KSK, PFL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3079 77 844 SAT-LB17 PO01_LB Saturday 783 2012


784 ENDO12L_SAT-LB18 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Angiotensin-II Activation of Na[sup]+[/sup]/H[sup]+[/sup] Exchanger-3 and mTORC1 in the Proximal Tubule Is Suppressed by Dipeptidyl Peptidase-4 Inhibitor Safwan Hyder, Chirag Mandavia, Irina Mugerfeld, Javad Habibi, Adam Whaley-Connell, James R Sowers, Ravi Nistala University of Missouri, Columbia, MO; University of Missouri, Columbia, MO; University of Missouri, Columbia, MO; University of Missouri, Columbia, MO; Harry S Truman Memorial Veterans[apos] Hospital, Columbia, MO [bold]Background and rationale[/bold]: Sodium homeostasis and blood pressure regulation is modulated by the effector peptide for renin angiotensin system (RAS), angiotensin-II (Ang-II), via activation of Na[sup]+[/sup]/H[sup]+[/sup] exchanger-3 (NHE3) in the kidney proximal tubule (PT). Excessive sodium retention and activation of RAS in obesity and diabetes is thought to contribute to hypertension. Recently, the role of nutrient sensor mTOR/S6K1 (mTORC1) has been implicated in hyperfiltration seen in early diabetes. Similarly, obesity is characterized by increased mTORC1 signaling and NHE3 activity. In addition, the incretin Glucagon-Like Peptide-1 (GLP-1) or Dipeptidyl Peptidase-4 (DPPIV) inhibition has been shown to decrease NHE3 activity and ameliorate hypertension. However, it is unknown how Akt-mTORC1 interacts with DPPIV enzyme and if nephron hypertrophy is related to NHE3 activity. Therefore, we hypothesized that DPPIV inhibition may suppress Ang II-mediated activation of mTORC1.[br][bold]Methods[/bold]: Opossum PT cells stably expressing rat Ang-II Type-1 receptor (AT1R) were starved in DMEM/F12 plus 0.1% FBS overnight and stimulated with Ang II (100nM) and pre-treated for 60min, with either MK-0626 (5[micro]M) or olmesartan (1[micro]M) or AG1478 (1[mu]M). Western blots, immunohistochemistry and enzyme inhibitor assays were used.[br][bold]Results[/bold]: Ang II and EGF stimulation of PTCs for 10min, activated mTORC1 and ERK1/2 as demonstrated by increased P-S473-Akt, P-S2448-mTOR, P-T389-S6K1 and P-T202/204-ERK respectively. Olmesartan completely blocks Ang II activation of mTORC1 and ERK while MK-0626 inhibition was partial. AG1478 inhibition of EGF effects was complete while MK-0626 had no effect. In addition, MK-0626 abrogated Ang II-mediated activation of DPPIV. Lastly, DPPIV bound to GLP1R and PI3-K/Akt pathway components possibly via PH domains.[br][bold]Conclusion[/bold]: These results suggest that NHE3 regulation (sodium and pH balance) and renal hypertrophy by Ang II may be via activation of DPPIV in the brush border of PTCs.[br][br]Sources of Research Support: Merck Pharmaceuticals RN, JH; NIH RO1 HL07301 and HL0107910 JRS; NIH R03 AG040638 AWC.[br][br]Nothing to Disclose: SH, CM, IM, JH, AW-C, JRS, RN 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3082 77 845 SAT-LB18 PO01_LB Saturday 784 2012


785 ENDO12L_SAT-LB19 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Mineralocorticoid Receptor Antagonism Markedly Improves Coronary Vascular and Cardiac Function Independent of Blood Pressure in the Zucker Obese Rat Shawn Bender, Vincent G DeMarco, Mona Garro, M Harold Laughlin, James R Sowers University of MIssouri, Columbia, MO; University of Missouri, Columbia, MO; University of Missouri, Columbia, MO; University of Missouri, Columbia, MO; University of Missouri, Columbia, MO; Harry S Truman Memorial Veterans[apos] Hospital, Columbia, MO Enhanced renin-angiotensin-aldosterone system (RAAS) activation contributes to vascular and cardiac dysfunction in states of obesity and insulin resistance (IR). Recent work suggests that this may involve activation of local tissue mineralocorticoid receptors (MR) and is independent of changes in blood pressure. Therefore, we examined the hypothesis that obesity and IR-associated coronary vascular and cardiac dysfunction would be attenuated by systemic MR antagonism with a low, blood pressure-independent dose of spironolactone. We utilized the Zucker obese (ZO) rat which manifests IR, vascular and cardiac dysfunction. Thirty wk old ZO rats were treated with spironolactone (1mg/kg/day, sc pellet) or placebo for 21 days. Compared to Zucker lean (ZL) rats, ZO rats displayed obesity, hyperglycemia and hyperinsulinemia that were unchanged in spironolactone-treated ZO rats (ZOSp). Blood pressure, determined by tail cuff, was not different across all groups. Surprisingly, circulating aldosterone concentrations tended to be reduced in ZO rats ([sim]40%) compared to ZL and were largely unchanged in ZOSp. Vasodilation of isolated, pressurized coronary arterioles to acetylcholine and insulin was reduced in ZO versus ZL and was restored in ZOSp. Paradoxically, coronary arterioles from ZOSp rats demonstrated reduced distensibility compared to ZL and ZO arterioles. Echocardiography revealed deteriorating cardiac function in ZO compared to ZL rats indicated by decreased myocardial performance index (MPI). This was associated with left ventricular hypertrophy and elevated cardiac reactive oxygen species (ROS) in ZO compared to ZL (p=0.06). Treatment with spironolactone improved cardiac function (reduced MPI) and reduced cardiac ROS while LV hypertrophy was principally unchanged. These results demonstrate pronounced coronary vascular and cardiac benefits of MR antagonism in the insulin resistant ZO rat despite reduced circulating aldosterone levels. Importantly, the benefits of MR blockade were independent of changes in blood pressure and highlight the significance of coronary vascular and cardiac MR in the pathogenesis of IR-associated cardiovascular disease.[br][br]Sources of Research Support: R01 HL 073101 and R01 HL0107910 JRS.[br][br]Nothing to Disclose: SB, VGD, MG, MHL, JRS 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3086 77 846 SAT-LB19 PO01_LB Saturday 785 2012


786 ENDO12L_SAT-LB20 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Local L-Type Ca2+ Channel Signaling in [alpha]T3-1 Cells Nathan L Chaplin, Christianne Magee, Dilyara Murtazina, Colin M Clay, Gregory C Amberg Colorado State University, Fort Collins, CO BACKGROUND: A dramatic release of luteinizing hormone (LH) from pituitary gonadotrope cells is necessary for ovulation. Binding of the hypothalamic decapeptide gonadotropin releasing hormone (GnRH) to its heterotrimeric Gq-protein coupled receptor on the gonadtrope cell surface initiates multiple signaling cascades, ultimately resulting in the release of LH and induction of ovulation. We hypothesized that the signaling molecule diacylglycerol, produced by phospholipase C, could activate protein kinase C and stimulation of L-type Ca2+ channels. To test this hypothesis we used a combination of TIRF microscopy and electrophysiology to image subplasmalemmal Ca2+ influx in the gonadtrope cell line [alpha]T3-1. Using this approach we visualized discrete sites of Ca2+ influx ([ldquo]Ca2+ sparklets[rdquo]) which produced microdomains of elevated Ca2+. METHODS: Ca2+ influx through L-type channels was imaged with the Ca2+ indicator fluo-5F (200 [mu]M). The external solution contained 2mM of Ca2+; K+ and Na+ currents were eliminated by replacing those ions with Cs+ and NMDG+, respectively. Experiments were performed at room temperature with cells voltage clamped at -70 mV. After establishing basal activity, GnRH (10 nM) was applied. For all experiments, cells were held at for 1 min prior to data acquisition and images were acquired at 50 Hz. Analysis was performed with custom software that included Ca2+ sparklet site density (sites/[mu]m2) and Ca2+ sparklet site activity (nPs; n is the number of quantal levels and Ps is the probability that the site is active). RESULTS: Following exposure of [alpha]T3-1 cells (n=3) to GnRH, sites of localized Ca2+ influx were identified, whereas a change in global Ca2+ were not evident. The L-type calcium channel blocker, nicardipine (10 [mu]M) abolished Ca2+ influx in response to GnRH (n=3). Conversely, the L-type Ca2+ channel agonist FPL64176 (500 nM) produced Ca2+ influx events indistinguishable from those induced by GnRH (n=3). CONCLUSIONS: These data indicate that in [alpha]T3-1 cells, GnRH activates L-type Ca2+ channels resulting in microdomains of elevated Ca2+. Interestingly, the Ca2+ signals in [alpha]T3-1 cells appear to be identical to those observed in arterial smooth muscle in response to the peptide hormone angiotensin II. Finally, the existence of Ca2+ microdomains in [alpha]T3-1 cells may explain the divergent signaling cascades produced by local vs. global Ca2+ events following GnRH receptor activation.[br][br]Sources of Research Support: NIH Grant 5R01HD065943 awarded to CMC.[br][br]Nothing to Disclose: NLC, CM, DM, CMC, GCA 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3015 77 847 SAT-LB20 PO01_LB Saturday 786 2012


787 ENDO12L_SAT-LB21 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Transcriptional Regulation of Human 5-Aminolevulinate Synthase 1 (ALAS1) Gene in Steroidogenic Cells Tetsuya Mizutani, Yunfeng Ju, Yoshitaka Imamichi, Takashi Yazawa, Takehiro Matsumura, Shinya Kawabe, Masafumi Kanno, Akihiro Umezawa, Kaoru Miyamoto University of Fukui, Eiheiji, Fukui, Japan; University of Fukui, Eiheiji, Fukui, Japan; National Research Institute for Child Health and Development, Setagaya, Tokyo, Japan Nuclear receptor 5A (NR5A) family proteins, steroidogenic factor-1 (SF-1) and liver receptor homolog-1 (LRH-1), play pivotal roles in regulation of steroidogenic enzymes. We have reported that bone marrow-derived mesenchymal stem cells (MSCs) can be differentiated into steroidogenic cells by the expression of SF-1/LRH-1. We used the differentiated MSCs to identify novel target genes for SF-1 by genome wide analyses, a promoter tiling array (ChIP-on-chip) and DNA microarray. We identified a novel candidate gene, ALAS1, that is an initiating and rate-limiting enzyme for mammalian heme biosynthetic pathway. Heme is essential for the catalytic activities of P450 enzymes including steroid metabolic enzymes, and therefore ALAS1 may be involved in steroid metabolism.[br]Chromatin immunoprecipitation and reporter assays revealed that SF-1/LRH-1 up-regulated ALAS1 gene transcription in steroidogenic cells, but not in hepatic cells, via binding to a 3.5 kb upstream region of ALAS1. ALAS1 gene expression was observed to be influenced by SF-1/LRH-1 in steroidogenic cells with overexpression of SF-1/LRH-1 and knockdown of SF-1 leading to up-regulated and down-regulated ALAS1 expression levels, respectively. Peroxisome proliferator-activated receptor-[gamma] coactivator-1[alpha] (PGC-1[alpha]), a co-activator of nuclear receptors, also strongly co-activated expression of SF-1/LRH-1-target genes. Reporter analysis revealed that PGC-1[alpha] strongly augmented ALAS1 gene transcription caused by SF-1 binding to the 3.5 kb upstream region. Finally knockdown of ALAS1 resulted in reduced progesterone production by steroidogenic cells. These results indicate that ALAS1 is a novel NR5A-target gene and participates in steroid hormone production.[br][br]Nothing to Disclose: TM, YJ, YI, TY, TM, SK, MK, AU, KM 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3022 77 848 SAT-LB21 PO01_LB Saturday 787 2012


788 ENDO12L_SAT-LB22 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Effect of Ghrelin on Chronic Liver Injury in Male Rats. Possible Role of Nitric Oxide Nashwa Negm Eldin Kabil, Hanan Ahmed Sedek, Nadia Aziz Yassin, Maha Mohamed Gamal German University in Cairo, Cairo, Egypt; Cairo University, Cairo, Egypt [bold]Background and Aims:[/bold] Recent studies have revealed that ghrelin may be an antioxidant and anti inflammatory agent in many organs in the body, however its role in chronic liver injury remains unclear. The role of NO in chronic liver injury is controversial as evidence suggests that NO is either a primary mediator of liver cell injury or posses a protective effect against injurious stimuli. Recent evidence demonstrated that the therapeutic potential for ghrelin was through eNOS activation and increase in NO production in metabolic syndrome. However, its role on NO production in the liver has not been previously investigated. The aim of this present study was designed to investigate the possible role of ghrelin in treatment of chronic liver injury. In addition, the possible mechanisms of action of Ghrelin in chronic liver injury and whether this action is mediated through nitric oxide. [bold]Methods:[/bold] Forty male rats were used in this study and were divided into the following four groups: Group 1: Control; Group 2: Chronic Liver Injury (CLI) (Thioacetamide 200mg/kg i.p for 6 weeks); Group 3: CLI [amp] Ghrelin (as Group 2 plus Ghrelin 10 ng/kg/day for 6 weeks); and Group 4: CLI + Ghrelin + L-NAME (as Group 3 plus L-NAME 20mg/kg/day orally for 6 weeks). Liver enzymes such as ALT (alanine aminotransferase) AST (aspartate aminotransferase), ALP (Alkaline Phosphatase) and Tumor necrosis factor alpha (TNF -[alpha]), were measured to assess liver function. In addition liver tissue collagen content, malondialdehyde (MDA), as well as gene expression of BAX, BCL2, and eNOS were assessed in order to determine the underlying mechanism of ghrelin action in the liver. [bold]Results:[/bold] Results showed that Ghrelin decreased serum liver enzymes, as well as TNF-[alpha] levels. In addition, Ghrelin reduced liver collagen, MDA, and pro apoptotic BAX gene expression, as well as increased antiapoptotic BCL2, and eNOS gene expression. These effects on TNF-[alpha], collagen, MDA, BAX, and eNOS were partially reversed in the TAA + Ghrelin + L-NAME group, suggesting that Ghrelin[apos]s effect on these parameters could be partially through modulation of NO levels. [bold]Conclusion:[/bold] Ghrelin has a partially protective effect against chronic liver injury which could be partially mediated by increasing NO levels.[br][br]Nothing to Disclose: NNEK, HAS, NAY, MMG 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3044 77 849 SAT-LB22 PO01_LB Saturday 788 2012


789 ENDO12L_SAT-LB23 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Atherogenic Dyslipidemia in Obese Adolescents and a Rodent Model of Polycystic Ovary Syndrome Ye Wang, Diane Abdoulaye, Geoff D Ball, Mary M Jetha, Donna F Vine University of Alberta, Edmonton, AB, Canada; University of Alberta, Edmonton, AB, Canada Introduction: Polycystic ovary syndrome (PCOS) is a metabolic-endocrine disorder associated with hyperandrogenemia, obesity and insulin resistance, predisposing individuals to increased risk of cardiovascular disease and Type 2 Diabetes. Dyslipidemia is a predominate feature of PCOS, however there remains limited data on the fasting and postprandial lipoprotein profile, and the relationship between hyperandrogenemia and atherogenic dyslipidemia.[br]Aim: The aim of this study was to determine 1) the fasting and postprandial metabolism of atherogenic lipids and lipoproteins, and 2) the correlation between these atherogenic biomarkers and hyperandrogenemia in obese adolescent girls with and without PCOS, and in an established rodent model of PCOS.[br]Methods and Results: Obese girls aged 14-18 years with or without PCOS were recruited (BMI [ge]95th percentile) and plasma lipid and lipoprotein (including apolipoproteins (apo) B100, a marker of hepatic lipoproteins (VLDL/LDL) and apoB48, a marker of intestinal chylomicrons) were measured during the fasted and postprandial state following a high fat meal. PCOS subjects had elevated fasting plasma TG, total and LDL-cholesterol, and lower HDL-cholesterol compared to obese girls without PCOS. Fasting plasma apoB48 and apoB100 concentrations were significantly elevated above pediatric reference values in both groups. The PCOS group had 40% higher fasting apoB100 and an exacerbated postprandial TG response to the high fat meal. The free androgen index was highly correlated with fasting (r=0.78, p[lt]0.0001) and postprandial TG response (r=0.5, p[lt]0.02).[br]In the JCR:LA-cp rodent model of PCOS fasting plasma TG, total cholesterol, apoB100 and apoB48 concentrations were elevated compared to control animals. Postprandial TG and apoB48 were elevated in response to a high fat meal in PCOS animals compared to controls, and serum total testosterone concentrations were correlated with fasting and postprandial TG (r=0.67, p[lt]0.01) and apoB48 (r=0.86, p[lt]0.001) response.[br]Conclusion: Collectively, this clinical and animal model data demonstrates exacerbated fasting and postprandial lipemia in PCOS, and this is associated with intestinal and hepatic apoB lipoproteins, and is positively correlated with hyperandrogenemia. These studies provide a rationale to assess the early fasting and postprandial dyslipidemic profile in adolescent PCOS and to further explore mechanisms of apoB associated dyslipidemia in the PCOS condition.[br][br]Nothing to Disclose: YW, DA, GDB, MMJ, DFV 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3059 77 850 SAT-LB23 PO01_LB Saturday 789 2012


790 ENDO12L_SAT-LB24 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) RF9, a Potent Antagonist of RFRP3 Receptor Signaling, Disinhibits Secretion of LH in the Seasonally Anovulatory Mare Jennifer Frances Thorson, Ligia Dias Prezotto, Alain Caraty, Marcel Amstalden, Gary Lynn Williams Texas A[amp]M University, Beeville, TX; Texas A[amp]M University, College Station, TX; Institut National de la Recherche Agronomique, Nouzilly, Centre, France Gonadotropin-inhibiting hormone (GnIH) was named for its ability to inhibit secretion of LH in birds and appears to contribute to reproductive seasonality in avian species. The mammalian homologue, RF-amide related peptide 3 (RFRP3), has also been implicated in regulation of LH secretion in rodents, sheep, and cattle, but results have been inconsistent. Although GnIH/RFRP3 appear to function through the G protein-coupled receptor 147 (GPR147; alias NPFFR1) expressed within both the hypothalamus and pituitary, functional variability associated with RFRP3 effects has resulted in uncertainty regarding its mechanism of action in mammals. In fact, in the mare, we have failed to demonstrate any effects of RFRP3 [italic]in vivo[/italic] or in adenohypophyseal cell culture. However, recently, a selective antagonist for the NPFF1 receptor has been shown to induce a potent release of LH in rats, mice, and sheep when administered centrally or peripherally, thus providing an additional approach for studying the RFRP3 system. The objective of the current experiment was to test the hypothesis that peripheral administration of RF9 can induce secretion of LH in the anovulatory mare nearing the time of vernal transition. On experimental day 0, 10 non-cyclic mares (n = 5/group) received three hourly intravenous injections of saline or RF9 (0.2 mg/kg BW) and jugular blood samples were collected at 10-min intervals for 5 h beginning 1 h before treatment. Mares were then treated continuously with GnRH (100 [micro]g/h) for 72 h via subcutaneous osmotic pumps, an approach uniquely effective in the equine for stimulating adenohypophyseal synthesis of LH. Beginning 12 h after pump removal (Day 4), mares were sampled at 10-min intervals for 6 h, with saline or RF9 (0.4 mg/kg BW) administered intravenously at 1 and 2 h. A GnRH challenge (250 [micro]g, iv) at hour 4 was conducted to test adenohypophyseal responsiveness. RF9 induced a robust and sustained increase ([italic]P[/italic] [lt] .0001) in circulating concentrations of LH relative to controls on Day 0. On Day 4, 3 of 5 ([italic]P[/italic] = 0.08) mares responded to RF9 treatment with an average 150% increase in circulating LH above pretreatment concentrations. Adenohypophyseal responsiveness to GnRH was not affected by RF9 treatment. Data are the first to illustrate functional modulation of the RFRP3 system in the horse and provide evidence that RF9 may act at the GnRH neuronal level to stimulate or disinhibit secretion of GnRH.[br][br]Sources of Research Support: Texas H-9137; The Link Equine Research Fund.[br][br]Nothing to Disclose: JFT, LDP, AC, MA, GLW 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3069 77 851 SAT-LB24 PO01_LB Saturday 790 2012


791 ENDO12L_SAT-LB25 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Obesity-Like Adipose Stroma Promotes Angiogenesis in a Humanized Mouse Model of Breast Cancer Lisa M Arendt, Jessica McCready, Patricia J Keller, Charlotte Kuperwasser Tufts University, Boston, MA Obesity has been identified as the most important risk factor associated with breast cancer in postmenopausal women. When compared with women of normal body mass index, obese women are more likely to be diagnosed with larger, higher grade tumors, an increased incidence of lymph node metastases, and elevated risk of distant recurrence. Given that obesity rates in the United States are rising, understanding the basis for breast cancer associated with obesity is of significant clinical relevance. Breast stromal tissue is a reservoir of subcutaneous fat, a fact that has been largely ignored in the study of breast cancer and may have a profound effect on breast cancer development and progression. Obese adipose tissue is associated with chronic inflammation and recruitment of proinflammatory macrophages, which is thought to occur in part due to increased expression of monocyte chemoattractant protein-1 (MCP-1) produced by activated adipocytes. However, the role of adipose tissue macrophages in obesity and cancer is largely unknown. We hypothesize that during obesity, adipose production of MCP-1 in cooperation with IL-1b, attracts and activates macrophages to the breast stroma, creating an environment in which premalignant lesions rapidly progress to aggressive tumors through enhanced angiogenesis and inflammatory signaling. In order to study the mechanisms of obesity in human breast cancer progression, we created a model derived from human breast reduction mammoplasty tissues in which obesity-like preadipocytes were created by overexpressing MCP-1 in primary breast preadipocytes. In the Human-In-Mouse model, obesity-like preadipocytes were found to promote higher grade breast cancers with a shorter latency and increase macrophage recruitment in preneoplastic lesions. In the absence of breast cancer cells, obesity-like preadipocytes significantly enhanced neoangiogenesis compared to control preadipocytes, and the angiogenic response was abrogated in the absence of macrophages. In vitro, macrophages stimulated with conditioned media from obesity-like preadipocytes significantly induced stromal-derived factor (SDF)-1a secretion which has been shown to enhance angiogenesis. SDF-1a induction was significantly reduced by inhibition of MCP-1 and IL-1b. These results suggest that reduction of localized inflammation within adipose tissue of the breasts may reduce angiogenesis providing high risk, obese women with increased therapeutic options.[br][br]Sources of Research Support: Raymond and Beverly Sackler Foundation, Breast Cancer[br]Research Foundation, Breast Cancer Alliance, NIH/NCI CA125554, CA092644, (C.K.), and NCRR K01-RR021858 (L.A.). C.K. is a Raymond and Beverly Sackler Foundation Scholar.[br][br]Nothing to Disclose: LMA, JM, PJK, CK 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3007 77 852 SAT-LB25 PO01_LB Saturday 791 2012


792 ENDO12L_SAT-LB26 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) A Forward Genetics Approach To Identify and Characterize Novel Gene Defects Associated with Genitourinary Birth Defects Shuo Han, Mounia Tannour-Louet, Jean-Francois Louet, Karina Romero, Josephine Addai, Dolores J Lamb Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX Congenital genitourinary (GU) abnormalities such as hypospadias, cryptorchidism and ambiguous genitalia are among the most common human birth defects. Care of the affected children is complicated by surgical, psychological, social and sexual concerns, which also presents challenges for their parents and physicians. Unfortunately the etiology of these human reproductive disorders remains poorly understood. Submicroscopic chromosomal anomalies underlie many genomic syndromes causing mental retardation, developmental delays and heart defects. We hypothesized that GU abnormalities result from similar chromosomal aberrations that cannot be detected by a routine karyotype. A clinically validated comparative genomic hybridization microarray revealed the presence of copy number variants (CNVs) in children born with GU abnormalities that evaded detection by high-resolution karyotype. Confirmed [italic]de novo[/italic] duplication and deletion events significantly associated with GU defects when compared to 8951 non-GU patient controls ([italic]P[/italic]=6.08x10[sup]-12[/sup]). Among these variants, a gain on Xq28 present in 2 unrelated patients encompassed a single gene; vesicle-associated membrane protein 7 ([italic]VAMP7[/italic]). Fluorescent in-situ hybridization confirmed the Xq28 gain and was a [italic]de novo[/italic] event. In the literature, several patients with GU anomalies displayed large karyotypically defined duplications in PAR2 of the X chromosome encompassing numerous genes, including VAMP7. To unravel the precise function of VAMP7 in the development of the genital tract, transgenic mice overexpressing VAMP7 were generated. The mice displayed unilateral or bilateral cryptorchidism with focal defects in spermatogenesis, reduced sperm motility and subfertility. Elevated levels of VAMP7 significantly reduced androgen receptor activity by trapping the receptor in the endosomal compartment, despite the presence of ligand. Increased levels of VAMP7 also markedly enhanced estrogen receptor [alpha] transcriptional activity. While [italic]in vivo[/italic] binding of androgen receptor decreased in VAMP7 mouse testis, expression of estrogen-responsive genes including ATF3, a variant implicated in human hypospadias, was upregulated in the testis and penile tissues of mutant mice. The identification of the chromosomal aberrations causing GU birth defects, such as VAMP7 duplication, will improve patient diagnosis and genetic counseling. Our long-term goal is to understand the molecular mechanisms underlying the pathogenesis of human genitourinary birth defects.[br][br]Sources of Research Support: Supported in part by grants 1R01DK078121 and K12 DK0083014 (KURe) from NIDDK.[br][br]Nothing to Disclose: SH, MT-L, J-FL, KR, JA, DJL 2012-06-23T13:30:00 Expo 2012-06-23T00:00:00 1899-12-30T13:30:00 3010 77 853 SAT-LB26 PO01_LB Saturday 792 2012


793 ENDO12L_OR10-1 ORAL SESSION: Glucose Homeostasis: Integrative [amp] Endocrine Metabolic Control (11:15 AM-12:45 PM) Brain-Derived Neurotrophic Factor Attenuates Diabetic Hyperglycemia Via a Central Effect To Suppress Hepatic Glucose Production in Rats with Uncontrolled Diabetes Miles E Matsen, Brent E Wisse, Joshua P Thaler, Gerald J Taborsky, Jr, Michael W Schwartz, Gregory J Morton University of Washington, Seattle, WA; Veterans Affairs Puget Sound Health Care System, Seattle, WA Rationale: In a rat model of uncontrolled insulin-deficient diabetes (uDM), central leptin administration fully normalizes hyperglycemia via a mechanism involving both reduced hepatic glucose production (HGP) and increased glucose uptake. As a first step to determine whether this leptin effect involves increased signaling by brain derived neurotrophic factor (BDNF), a downstream target of leptin action in the brain, we investigated whether central administration of BDNF mimics leptin[apos]s anti-diabetic effects. Methods: Following cannulation of the lateral ventricle, adult male Wistar rats received systemic injection of either vehicle or streptozotocin (STZ; 40mg/kg/sc x2) to induce uDM. Four days later, rats received consecutive daily intracerebroventricular (icv) injections of BDNF (5ug/d) or vehicle (sterile water) for 7 days to create 3 groups: 1) veh-veh, 2) STZ-veh and 3) STZ-BDNF. To control for the effect of BDNF to reduce food intake, an additional group of STZ-veh animals were pair-fed to the intake of the STZ-BDNF group. Results: Daily icv administration of BDNF lowered blood glucose levels (STZ-BDNF: 261[plusmn]24 mg/dl vs. STZ-veh: 397[plusmn]10 mg/dl; p[lt]0.05) and prevented diabetic hyperphagia (STZ-BDNF: 21.7[plusmn]1.6g vs. STZ-veh: 41.8[plusmn]1.9 g vs. veh-veh: 27.5[plusmn]0.8g; p[lt]0.05), but the decrease in food intake could not account for the blood glucose reduction (STZ-veh-PF: 342[plusmn]12 mg/dl, p[lt]0.05 vs. STZ-BDNF). Instead, BDNF lowered blood glucose levels by potently suppressing hepatic glucose production (HGP) (mean HGP: 17.5[plusmn]6.4 mg/kg/min for STZ-BDNF vs. 35.6[plusmn]6.0 mg/kg/min for STZ-veh-PF; P[lt]0.05) without any effect on glucose uptake in peripheral tissues. While uDM characteristically increases hepatic expression of gluconeogenic genes (G6Pase and Pepck) and plasma glucagon levels, both were normalized by BDNF treatment (p[lt]0.05 for both). Taken together, these data suggest that in uDM 1) icv BDNF mimics the effect of leptin to lower blood glucose levels via reduced HGP, possibly through suppression of glucagon secretion and 2) leptin[apos]s effect to increase peripheral glucose utilization likely involves a mechanism distinct from BDNF signaling.[br][br]Sources of Research Support: NIH Grants: DK-089056(GJM) and DK-083042(MWS).[br][br]Nothing to Disclose: MEM, BEW, JPT, GJT, MWS, GJM 2012-06-24T11:15:00 320 2012-06-24T00:00:00 1899-12-30T11:15:00 415 146 967 OR10-1 OR14-01 Sunday 793 2012


794 ENDO12L_OR10-2 ORAL SESSION: Glucose Homeostasis: Integrative [amp] Endocrine Metabolic Control (11:15 AM-12:45 PM) Hepatic Rictor Gene Deletion Protects Against Development of Diet-Induced Obesity and Uncouples Dyslipidemia from Hyperglycemia Jennifer M Rojas, Richard L Printz, Kevin D Niswender Vanderbilt University School of Medicine, Nashville, TN; Vanderbilt University School of Medicine, Nashville, TN; VA Tennessee Valley Healthcare System, Nashville, TN Insulin is a key metabolic regulator of liver glucose and lipid metabolism, and selective hepatic insulin resistance has been implicated in the pathogenesis of hyperglycemia and dyslipidemia associated with obesity, diabetes and metabolic syndrome. AKT is an insulin signaling intermediary that regulates many of the metabolic actions of insulin in liver. AKT is activated via phosphorylation at Serine 473 (pS473) by the Rictor containing complex, mTORC2, in addition to PDK1 phosphorylation at Threonine 308 (pT308). We hypothesized that hepatic Rictor gene deletion using CRE-Lox P (Albumin-Cre) technology in mice would lead to partial AKT activation (pT308 but not pS473), creating an animal model of selective hepatic insulin resistance. Indeed, when this strategy was implemented, a gene dose dependent reduction in Rictor mRNA expression was observed; and in chow-fed null mice compared to controls, a 90% reduction in pS473-AKT was achieved, while maintaining pT308-AKT levels (83%) in liver. Although at 12 weeks of age, these chow-fed Rictor null mice had no significant alteration in body composition, glucose tolerance (as measured by IPGTT in 4-h fasted mice) or lipids. We next challenged the mice by feeding them for 10 weeks a high fat diet (HFD; 45% lard based diet) to determine whether the stress of diet-induced obesity (DIO) would exaggerate the phenotype. Surprisingly, despite Rictor null mice (KO) having similar food intake and lean mass as control mice (CNT) on HFD, we found that hepatic Rictor gene deletion prevented the development of DIO, as evidenced by 57% less fat mass gain (3.0[plusmn]0.8g for KO vs. 6.8[plusmn]0.6g for CNT, [italic]P[/italic][lt]0.01), 34% lower body weight gain (6.3[plusmn]0.9g for KO vs. 9.6[plusmn]0.6g for CNT,[italic] P[/italic][lt]0.01), and lower weights of epididymal fat pad (1.1[plusmn]0.2g for KO vs. 2.2[plusmn]0.1g for CNT,[italic] P[/italic][lt]0.001) and liver (0.7[plusmn]0.04g for KO vs. 1.2[plusmn]0.06g for CNT,[italic] P[/italic][lt]0.0001). Null mice, however, became severely glucose intolerant after 7 weeks of HFD feeding (area under the curve 35,921[plusmn]2073 for KO vs. 23,802[plusmn]1291 for CNT, [italic]P[/italic][lt]0.001). Strikingly, null mice on HFD fail to develop hepatic steatosis and dyslipidemia, as evidenced by 55% less liver triglyceride (TG; 14[plusmn]2.2[micro]g/mg for KO vs. 32[plusmn]2.6[micro]g/mg for CNT, [italic]P[/italic][lt]0.001) and 23% lower plasma TG (36[plusmn]3.4mg/dl for KO vs. 47[plusmn]2.5mg/dl for CNT, [italic]P[/italic][lt]0.05). In conclusion, targeted Rictor gene deletion in liver uncouples dyslipidemia from hyperglycemia in DIO mice protecting against the development of hepatic steatosis and hyperlipidemia.[br][br]Sources of Research Support: Resources of the Tennessee Valley Healthcare System, VA Merit Review, 1R01 DK085712-01 and 1F31DK089906-01.[br][br]Nothing to Disclose: JMR, RLP, KDN 2012-06-24T11:30:00 320 2012-06-24T00:00:00 1899-12-30T11:30:00 719 146 968 OR10-2 OR14-01 Sunday 794 2012


795 ENDO12L_OR10-3 ORAL SESSION: Glucose Homeostasis: Integrative [amp] Endocrine Metabolic Control (11:15 AM-12:45 PM) Metabolic Actions of FGF21 in Mice with Liver-Specific Insulin Resistance Brice Emanuelli, Sara Vienberg, Graham Smyth, Christine Cheng, Mervyn D Michael, Alexei Kharitonenkov, C Ronald Kahn Joslin Diabetes Center, Boston, MA; Lilly Corporate Center, Indianapolis, IN Obesity-associated metabolic complications including type 2 diabetes represent growing health problems worldwide. FGF21 is a novel metabolic regulator of glucose and lipid homeostasis, as well as body weight which could serve as new therapy for these disorders. Several lines of evidence suggest that FGF21 regulates hyperglycemia, at least in part, via the sensitization of insulin action, particularly in liver. However, exactly which tissues and to what extent insulin sensitivity contribute to FGF21 efficacy, remain to be determined.[br]To assess the role of liver insulin signaling in FGF21 metabolic actions we have used mice with liver selective inactivation of the insulin receptor (LIRKO mice). Male control and LIRKO animals were fed either chow or high fat diets for 7 weeks and treated with 1 [mu]g/g/day FGF21 or saline for the 2 last weeks of the diet using osmotic pumps. FGF21 increased energy expenditure equally as measured in metabolic cages in both control and LIRKO mice, without any change in activity or feeding behavior. This was accompanied by a reduction in body weight by FGF21 in both groups of mice, but no obvious change in inflammatory markers in fat.[br]Remarkably, despite absence of insulin signaling in the liver and the inability of FGF21 to reduce gluconeogenesis in LIRKO mice, hyperglycemia in LIRKO mice was totally normalized upon FGF21 treatment. Using 14C-2-deoxyglucose uptake in vivo to determine glucose fate, the reduced glucose levels were due, at least in part, to increased glucose uptake in brown fat. The marked hyperinsulinemia of LIRKO animals was also normalized by FGF21, and insulin tolerance was improved. On the other hand, the ability of FGF21 to lower plasma triglycerides, cholesterol, or FFAs, which was observed in control mice, was completely abolished in LIRKO mice, indicating that the lipid lowering effect of FGF21 requires liver insulin signaling. Consistent with this, the ability of FGF21 to control the expression of key genes involved in the regulation of glucose, lipid and cholesterol metabolism was altered in livers from LIRKO mice.[br]Together, our data show that FGF21 is able to exert its glucose lowering effect and correct hyperglycemia in mice which lack insulin action in liver. This occurs despite absence of effect on hepatic gluconeogenesis primarily through increased energy metabolism in brown fat. FGF21 action to control lipid metabolism, however, is dependent on insulin action in liver.[br][br]Disclosures: CC: Employee, Eli Lilly & Company. MDM: Investigator, Eli Lilly & Company. AK: Investigator, Eli Lilly & Company. Nothing to Disclose: BE, SV, GS, CRK 2012-06-24T11:45:00 320 2012-06-24T00:00:00 1899-12-30T11:45:00 1793 146 969 OR10-3 OR14-01 Sunday 795 2012


796 ENDO12L_OR10-4 ORAL SESSION: Glucose Homeostasis: Integrative [amp] Endocrine Metabolic Control (11:15 AM-12:45 PM) Structural Basis of Idealized Selective PPAR[gamma] Modulation Senapathy Rajagopalan, Angelica A Amato, Bruno Melo Carvalho, Ana Carolina M Figueira, Stephen D Ayers, Melina Mottin, Rodrigo L Silveira, Paulo CT Souza, Rosa HV Mourao, Mario JA Saad, Carl W Carruthers, Marie Togashi, Luiz A Simeoni, Dulcineia SP Abdalla, Munir S Skaf, Igor Polikparpov, Maria CA Lima, Suely L Galdino, Richard G Brennan, John D Baxter, Ivan R Pitta, Paul Webb, Francisco AR Neves, Kevin J Phillips The Methodist Hospital Research Institute, Houston, TX; Universidade de Bras[iacute]lia, Bras[iacute]lia, Brazil; Universidade Estadual de Campinas, S[atilde]o Paulo, Brazil; Brazilian Association for Synchroton Light Technology, S[atilde]o Paulo, Brazil; Universidade de Campinas, S[atilde]o Paulo, Brazil; Instituto Nacional de Ci[ecirc]ncia e Tecnologia para Inova[ccedil][atilde]o Farmac[ecirc]utica, Pernambuco, Brazil; Departamento de F[iacute]sica e Universidade de S[atilde]o Paulo, S[atilde]o Carlos, Brazil; Duke University School of Medicine, Durham, NC The recent discovery that PPAR[gamma] targeted antidiabetic drugs function by inhibiting Cdk5-mediated phosphorylation of the receptor has provided a new viewpoint to evaluate and perhaps develop improved insulin-sensitizing agents.[br]Herein we report the development of novel thiazolidinediones that retain similar anti-diabetic efficacy as rosiglitazone in mice, yet do not elicit weight gain or edema, common side effects associated with full PPAR[gamma] activation. Further characterization of one these compounds show GQ-16 to be an effective inhibitor of Cdk5-mediated phosphorylation of PPAR[gamma]. The structure of GQ-16 bound to PPAR[gamma] demonstrates that the compound utilizes a binding mode distinct from other reported PPAR[gamma] ligands, although it does share structural features with other partial agonists such as MRL-24 and PA-082 that have similarly been reported to dissociate insulin sensitization from weight gain. Hydrogendeuterium exchange studies reveal that GQ-16 strongly stabilizes the [beta]-sheet region of the receptor, presumably explaining the compounds efficacy in inhibiting Cdk5-mediated phosphorylation of S273. Molecular dynamics simulations suggest the partial agonist activity of GQ-16 indicating that the compound prevents helix 12 from docking efficiently in the active conformation. Our results suggest that the emerging model, whereby [apos]ideal[apos] PPAR[gamma]-based therapeutics stabilize the [beta]-sheet/S273 region and inhibit Cdk5-mediated phosphorylation while minimally invoking adipogenesis and classical agonism, is indeed a valid framework to develop improved PPAR[gamma] modulators that retain antidiabetic actions while minimizing untoward effects.[br][br]Nothing to Disclose: SR, AAA, BMC, ACMF, SDA, MM, RLS, PCTS, RHVM, MJAS, CWC, MT, LAS, DSPA, MSS , IP, MCAL, SLG, RGB, JDB, IRP, PW, FARN, KJP 2012-06-24T12:00:00 320 2012-06-24T00:00:00 1899-12-30T12:00:00 1838 146 970 OR10-4 OR14-01 Sunday 796 2012


797 ENDO12L_OR10-5 ORAL SESSION: Glucose Homeostasis: Integrative [amp] Endocrine Metabolic Control (11:15 AM-12:45 PM) Crosstalk between Liver and Muscle Causes Systemic Insulin Resistance in Mice with Liver-Specific Ablation of PPAR[gamma] Anisha A Gupte, Laurie J Minze, Zheng Yin, Jessica Martinez, Alan Collins, Christopher J Lyon, Willa A Hsueh The Methodist Hospital Research Institute, Houston, TX Fatty liver is associated with insulin resistance and diabetes. Although the role of peroxisome proliferator-activated receptors-[gamma] (PPAR[gamma]) in adipose tissue has been extensively studied, very little is known about the role of liver-PPAR[gamma] and its effects on liver metabolism. We thus generated mice with liver-specific PPAR[gamma] knockout (L-PPAR[gamma]KO), and found that L-PPAR[gamma]KO mice fed obesogenic high fat diet (HFD) for 3 months had elevated fasting insulin and systemic insulin resistance compared to wild type (WT) littermates and were more insulin resistant when assessed by insulin tolerance test. However, there were no differences in plasma triglycerides, phospholipids or free fatty acid levels. Insulin-stimulated Akt phosphorylation was increased in liver but reduced in skeletal muscle of HFD-fed L-PPAR[gamma]KO mice vs. WT mice. L-PPAR[gamma]KO mice had 50% the liver fat of WT mice. Mitochondrial function analyses revealed significant suppression of b-oxidation in the livers of L-PPAR[gamma]KO mice, but there was reduced expression of lipogenic genes (e.g. fatty acid synthase) corresponding with reduced hepatic fat accumulation, suggesting that liver-PPAR[gamma] directly regulates hepatic lipid synthesis and metabolism. However, the PPAR[gamma]-agonist rosiglitazone restored [beta]-oxidation in L-PPAR[gamma]KO mice on HFD to that seen in WT mice. Microarray analyses of L-PPAR[gamma]KO and WT livers revealed a marked suppression of several secreted factors, including the secreted complement factor D (adipsin), verified by quantitative real time PCR analyses. These data suggest: 1) liver PPAR[gamma] directly regulates hepatic lipid synthesis and mitochondrial [beta]-oxidation, 2) liver PPAR[gamma] may exert significant effects upon skeletal muscle insulin sensitivity, even in the absence of liver fat, via non-lipid secreted factors, and 3) reduced secretion of adipsin may contribute to skeletal muscle insulin resistance. We conclude that liver PPAR[gamma] may have important therapeutic implications for liver complications found in obesity and diabetes.[br][br]Sources of Research Support: American Heart Association. McDonald Foundation.[br][br]Nothing to Disclose: AAG, LJM, ZY, JM, AC, CJL, WAH 2012-06-24T12:15:00 320 2012-06-24T00:00:00 1899-12-30T12:15:00 2082 146 971 OR10-5 OR14-01 Sunday 797 2012


798 ENDO12L_OR10-6 ORAL SESSION: Glucose Homeostasis: Integrative [amp] Endocrine Metabolic Control (11:15 AM-12:45 PM) Bone as a Site of Insulin Resistance in Type 2 Diabetes Jianwen Wei, Gerard Karsenty Columbia University, New York, NY Type 2 diabetes is characterized by insulin resistance in various insulin target cells, the classical ones being the hepatocytes, myoblasts and adipocytes. The recent realization that the osteoblast is an insulin target cell involved in the control of whole-body glucose homeostasis suggests the possibility that impaired insulin signaling in osteoblasts contribute to the development of type II diabetes. To test whether it is the case, we first showed that insulin resistance occurred in osteoblasts in wild type mice rendered diabetic through a high-fat-diet (HFD). Molecular evidence of insulin resistance includes reduced phosphorylation of AKT at Th308 and Ser347 in bones, a decrease of bone resorption due to an increase of Opg expression and a decrease of the active form of osteocalcin. Next, to test If insulin resistance in osteoblasts does contribute to the systematic insulin resistance, we used loss-of (InsRosb+/- mice) and gain-of function ([alpha]1(I)-InsR transgenic mice) models of insulin signaling in osteoblasts. Glucose tolerance tests and insulin tolerance tests showed that InsRosb+/- mice were more severely glucose intolerant and insulin resistant than control mice fed the same HFD, while [alpha]1(I)-InsR mice were partially protected from insulin resistance. Accordingly, ex vivo cell culture experiments and in vivo hyperinsulinemic euglycemic clamp experiments showed that osteoblasts uptake glucose in an insulin-regulated manner and that decreased glucose uptake in osteoblasts is observed in mice fed a HFD. Lastly, gene expression analysis identified Glut1 as the main glucose transporter in osteoblasts controlling both basal and insulin-dependent glucose uptake. Our investigations of a mouse model lacking Glut1 only in osteoblasts will be presented at the meeting. Taken together, this study provided evidence that insulin resistance takes place in osteoblasts in diet-induced type 2 diabetes model.[br][br]Sources of Research Support: NIH Grant 5R01DK078042-04 to GK.[br][br]Nothing to Disclose: JW, GK 2012-06-24T12:30:00 320 2012-06-24T00:00:00 1899-12-30T12:30:00 1139 146 972 OR10-6 OR14-01 Sunday 798 2012


799 ENDO12L_OR11-1 ORAL SESSION: Novel Mechanisms for Testicular Function [amp] Male Reproductive Tract Development (11:15 AM-12:45 PM) E-Type Cyclins: Novel Key Regulators of Homologous Pairing, Synapsis and Chromosome Stability during Mammalian Spermatogenesis Laetitia Martinerie, Marcia Manterola, Sunil Panigrahi, Ana Vasileva, Melissa Weisbach, Yan Geng, Peter Sicinski, Debra Wolgemuth Columbia University Medical Center, New York, NY; Dana-Farber Cancer Institute, Boston, MA; Columbia University Medical Center, New York, NY; Columbia University Medical Center, New York, NY; Columbia University Medical Center, New York, NY Despite improvment in medical knowledge, infertility remains a concern for one in seven couples. Male factor infertility accounts for one third of infertile couples and most causes are poorly understood, including unexplained sperm deficiencies which are responsible for 30 % of male infertility.[br]E-type cyclins (E1 and E2) are key regulators of the mitotic cell cycle. Interestingly, while E1 or E2 deletion does not have any consequence in terms of development and life span, E2 knockout male mice have a reduced fertility. To better understand the role of E-type cyclins during spermatogenesis, we first characterized their expression in the testis at the mRNA and protein levels. We observed a surprising discordance between E1 mRNA (present in spermatogonia and meiotic cells), and protein expression (limited to spermatogonia only). Cyclin E2 mRNA and protein expression was more narrowly restricted, being detected solely in spermatocytes from late zygotene to early diplotene. The sequential deletion of E1 and E2 alleles resulted in increased infertility, with E1[sup]+[/sup]/[sup]+[/sup]E2[sup]-[/sup]/[sup]- [/sup]male mice exhibiting subfertility to complete infertility in E1[sup]+[/sup]/[sup]-[/sup]E2[sup]-[/sup]/[sup]-[/sup] male mice. Interestingly, in the E1[sup]+[/sup]/[sup]+[/sup]E2[sup]-[/sup]/[sup]-[/sup] male mice, E1 protein expression was now detected in meiotic spermatocytes, perhaps compensating in part for the loss of E2 function and abrogating the severity of the phenotype. A testis-specific complete loss of cyclin E1 and E2 function was achieved by a combination of matings using the knockout alleles and a conditional knockout allele of cyclin E1 combined with Stra8-Cre mice. These male mice were completely infertile and exhibited an increased apoptosis of pachytene spermatocytes. This apoptosis was secondary to an abnormal loading of synaptonemal complex components. In addition, there were heterologous chromosome associations and numerous unrepaired double strand breaks (DSBs) during late stages of prophase as revealed by abnormal foci of gH2AX and SUMO-1. Moreover, several foci were observed in the telomeres, concomitant with a disruption in telomeric DNA organization, suggesting that E type cyclins are fundamental for the telomere stability.[br]These results uncover a new role for E-type cyclins, highlighting them as key regulators of spermatogenesis, being crucial for meiotic prophase 1 spermatocyte progression and stability. These observations open an exciting new area of research in heretofore uncharacterized human male infertility.[br][br]Nothing to Disclose: LM, MM, SP, AV, MW, YG, PS, DW 2012-06-24T11:15:00 362 2012-06-24T00:00:00 1899-12-30T11:15:00 2206 147 973 OR11-1 OR35-01 Sunday 799 2012


800 ENDO12L_OR11-2 ORAL SESSION: Novel Mechanisms for Testicular Function [amp] Male Reproductive Tract Development (11:15 AM-12:45 PM) Regulation of Spermatogenesis by the Shp2 Phosphatase Pawan Puri, William H Walker University of Pittsburgh, Pittsburgh, PA Spermatogenesis and male fertility requires maintenance of the spermatogonial stem cell (SSC) pool and integrity of the blood-testis barrier (BTB), a cell-junctional complex made between Sertoli cells in the testis. Protein tyrosine phosphatase 11 (PTPN11/SHP2) is a non-receptor PTP that regulates pathways essential for the renewal and survival of hematopoietic stem cells as well as maintaining the integrity of epithelial-cell-junctions. The gain and loss of function SHP2 mutations result in the genetic diseases Noonan syndrome and LEOPARD syndrome, respectively that have numerous associated pathologies including male infertility. In other cell types, SHP2 transduces signals from glial cell derived neurotrophic factor (GDNF) and hepatocyte growth factor (HGF), two growth factors that are essential for the spermatogonial stem cell renewal and maintaining BTB integrity. We found that in testis, SHP2 is expressed in Sertoli cells at the BTB as well as in SSCs and differentiated spermatogonia. We found that SHP2 regulates the activity of kinases and the localization of junctional proteins in Sertoli cells that are required to maintain the BTB. In studies of Sertoli cells cultured from 20 day-old rats, we found that SHP2 is activated by HGF and epidermal growth factor and SHP2 activates the MAP kinase pathway via Src kinase. SHP2 also directly interacts with and inactivates FAK kinase. Constitutive activation of SHP2 causes actin depolymerization in Sertoli cells. Also, Sertoli cell proteins required for maintaining the BTB are mis-localized away from the plasma membrane after constitutive activation of SHP2. Finally, introduction of constitutively active SHP2 into Sertoli cells results in the loss of tight junction integrity that is required to maintain the BTB. We discovered that inducible ablation of SHP2 in adult mice causes disruption of spermatogenesis due to inability of undifferentiated spermatogonia/SSCs to replenish the seminiferous tubules with germ cells. Therefore, we hypothesize that SHP2 is required in both Sertoli cells and developing germ cells to maintain spermatogenesis and male fertility. Studies are underway to identify the specific functions and mechanism of action of SHP2 in Sertoli cells and germ cells using cell specific conditional knock out mice. Our studies show that SHP2 function is essential for spermatogenesis and may define the pathogenesis of male infertility in Noonan and LEOPARD syndrome patients.[br][br]Sources of Research Support: This work is supported by fellowships from the Lalor Foundation and Magee Womens Research Institute to PP.[br][br]Nothing to Disclose: PP, WHW 2012-06-24T11:30:00 362 2012-06-24T00:00:00 1899-12-30T11:30:00 1655 147 974 OR11-2 OR35-01 Sunday 800 2012


801 ENDO12L_OR11-3 ORAL SESSION: Novel Mechanisms for Testicular Function [amp] Male Reproductive Tract Development (11:15 AM-12:45 PM) GnRH Antagonist Treatment Promotes Spermatogenesis from Endogenous and Autologously Transplanted Spermatogonia in Cynomolgus Monkeys Gunapala Shetty, Shan H Shao, Rajesh K Uthamanthil, Wei Zhou, Connie C Weng, Brian P Hermann, Kyle E Orwig, Marvin L Meistrich University of Texas MD Anderson Cancer Center, Houston, TX; University of Pittsburgh School of Medicine, Magee-Womens Research Institute, Pittsburgh, PA Cancer treatment regimens can completely kill the stem spermatogonia in children and young patients in which case cryopreservation of spermatogonia before therapy and subsequent autologous transplantation may be the only option for restoring fertility. To optimize conditions for this strategy, we irradiated the testes of 12 cynomolgus monkeys with 7 Gy and 2 months later transplanted cryopreserved testicular cells, marked by lentivirus GFP transduction, autologously back to one of the testes. As we demonstrated earlier that testosterone (T) suppression enhanced colonization and differentiation of transplanted spermatogonia in rodents, half of the irradiated monkeys were treated with GnRH antagonist (GnRH-ant) starting immediately after irradiation until the time of transplantation. Several parameters indicated that GnRH-ant Rx stimulated endogenous spermatogenic recovery: At 11 months after irradiation, the testicular volume of untransplanted testes in the GnRH-ant-treated monkeys, (42% of pre-Rx value) was higher than that of the radiation-only monkeys (33% of pre-Rx value). At 6.5 months after irradiation the untransplanted testes in the GnRH-ant-treated monkeys showed differentiated germ cells in 11% of tubule sections compared to 2.5% in the irradiated-only monkeys; at 11 months after irradiation these numbers increased to 13% and 7% respectively.[br]GnRH-ant Rx also enhanced spermatogenic recovery from the transplanted cells: The transplanted testes in the GnRH-ant-treated monkeys weighed 20% more than the contralateral testes; no such weight differentials were observed in irradiated-only transplanted testes. Five out of six GnRH-ant-treated, transplanted monkeys consistently showed sperm in the semen ([lt] 9 x 10[sup]6[/sup]/ejaculate) after transplantation, compared to 2/6 monkeys in irradiated-only group ([lt] 0.2 x 10[sup]6[/sup]/ejaculate). GnRH-ant-treated irradiated monkeys showed differentiated germ cells in 17% of tubules in the transplanted testes compared to only 13% in the contralateral untransplanted testis. Post-transplantation sperm from 4 monkeys from the GnRH-ant-treated group showed lentivirus/GFP DNA confirming recovery from the transplanted cells, but sperm from only 1 irradiated-only transplanted monkey was positive for lentivirus/GFP. These studies indicate that T suppression enhances spermatogenic recovery from both endogenous and transplanted stem spermatogonia in primates and can be a useful tool in the restoration of fertility after cancer treatment in men.[br][br]Sources of Research Support: NIH Grant R21-HD061301 awarded to GS.[br][br]Nothing to Disclose: GS, SHS, RKU, WZ, CCW, BPH, KEO, MLM 2012-06-24T11:45:00 362 2012-06-24T00:00:00 1899-12-30T11:45:00 1066 147 975 OR11-3 OR35-01 Sunday 801 2012


802 ENDO12L_OR11-4 ORAL SESSION: Novel Mechanisms for Testicular Function [amp] Male Reproductive Tract Development (11:15 AM-12:45 PM) Role of E2F1 in Testicular Descent and Fertility Carolina J Jorgez, Venkata H Vangapandu, Aysegul Sahin, Jill A Rosenfeld, Dolores J Lamb Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; PerkinElmer, Inc, Spokane, WA E2F1 is a potent transcription factor that activates the cell cycle by regulating expression of genes required for DNA synthesis and cell proliferation. Mice lacking [italic]e2f1[/italic] are viable and initially fertile, yet experience testicular atrophy with age, exocrine gland dysplasia and a spectrum of tumors. Testicular atrophy can be seen as early as 12 weeks due to spermatogonial cell loss. On the other hand, mice overexpressing [italic]e2f1[/italic] are infertile with testicular atrophy from p53-independent apoptosis occurring within individual tubules that results in germ cell but not Sertoli cell depletion. Our group discovered that infertile men with microduplication or microdeletion, as well as missense mutations in [italic]E2F1,[/italic] display abnormal spermatogenesis that correlates with the type of genomic change. Since humans lacking or with an extra copy of [italic]E2F1 [/italic]are infertile, we more closely define the phenotype of [italic]e2f1[/italic] deficient mouse. The mice are subfertile with a decrease in the number of litters per month, although the number of pups per litter was not different. By 6 weeks of age there is a significant decrease in testis weight, sperm count and motility. However, the most surprising finding is that these mice have unilateral inguinal cryptorchidism. Sixty patients with cryptorchidism were analyzed for the presence of copy number variations (CNVs) in [italic]E2F1[/italic] by qPCR using Taqman CNV assays. Three cryptorchid patients displayed CNVs in [italic]E2F1[/italic] whereas none of 58 normal men did. A search of the Signature Genomic Laboratories database of 26,706 individuals tested by array comparative genomic hybridization revealed two young males with microduplication in 20q that includes [italic]E2F1[/italic], with one of these having an undescended right testis (no GU information on the other boy was available). Analysis of the DECIPHER database yielded 2 boys with 20q microduplications encompassing E2F1, one of whom had cryptorchidism (5404 subjects are deposited in this database, no GU information on the other boy was available). Taken together, our data suggest a role of [italic]E2F1[/italic] not only in spermatogenesis but also in testicular descent.[br][br]Sources of Research Support: This study was supported in part by NIH grants K12 DK0083014 (KURe) and R01DK078121 from NIDDK and P01HD36289 from NICHD. This study makes use of data generated by the DECIPHER Consortium. A full list of centers who contributed to the generation of the data is available from http://decipher.sanger.ac.uk and funding for the project was provided by the Wellcome Trust.[br][br]Nothing to Disclose: CJJ, VHV, AS, JAR, DJL 2012-06-24T12:00:00 362 2012-06-24T00:00:00 1899-12-30T12:00:00 2039 147 976 OR11-4 OR35-01 Sunday 802 2012


803 ENDO12L_OR11-5 ORAL SESSION: Novel Mechanisms for Testicular Function [amp] Male Reproductive Tract Development (11:15 AM-12:45 PM) Deletion of a Novel lncRNA Results in Severe Hypospadias Paul E Gradie, Paul Overbeek, Richard R Behringer, Andrew J Pask University of Connecticut, Storrs, CT; Baylor College of Medicine, Houston, TX; University of Texas MD Anderson Cancer Center, Houston, TX Hypospadias (the abnormal placement of the urethral opening on the penis) is one of the most common birth defects in the USA, affecting approximately 1 in every 140 live male births. Despite its high incidence, relatively little is known about the causes of this common disease. To date, only a few genes have been isolated that predispose the penis to hypospadias. Here we report on the identification of a novel long non-coding RNA molecule that is required for urethral closure. Deletion of this gene results in severe penoscrotal hypospadias phenotype with undescended testes and an unfused scrotum. While the urethral epithelium appears to form normally and is present along the entire ventral surface of the phallus, the urethra fails to close. A defect in urethral closure is apparent as early as E15.5, and mutants can be visually distinguished on the day of birth. In addition, females deleted for this gene also displayed urogenital abnormalities and reduced anogenital distance. We have determined that the novel long non-coding RNA molecule (LncA) is located approximately 230kb downstream of the EphrinB2 gene. LncA is expressed throughout the urethral closure window and alongside EphrinB2. The proximity of LncA to EphrinB2 and their overlapping phenotypes suggests LncA may mediate EphrinB2 function. Furthermore LncA is surrounded by multiple functional estrogen- and androgen-receptor binding sites suggesting it may be a direct target of endocrine disruption in the etiology of hypospadias.[br][br]Nothing to Disclose: PEG, PO, RRB, AJP 2012-06-24T12:15:00 362 2012-06-24T00:00:00 1899-12-30T12:15:00 141 147 977 OR11-5 OR35-01 Sunday 803 2012


804 ENDO12L_OR11-6 ORAL SESSION: Novel Mechanisms for Testicular Function [amp] Male Reproductive Tract Development (11:15 AM-12:45 PM) CAR Knockout Mice Have Decreased Systemic Androgen Levels Leading to Hypertrophy of the Testes, and Atrophy of the Prostate and Seminal Vesicles Juan Pablo Hernandez, Sayee Anakk, David D Moore Baylor College of Medicine, Houston, TX The constitutive androstane receptor[apos]s (CAR) regulation of detoxification pathways have been extensively studied, and recently CAR[apos]s role in intermediary metabolism have also been established as well. Detoxification of endogenous compounds, for example bilirubin, bile acids, and thyroid hormone, due to CAR activation have been established; however, activation of CAR by estradiol and inactivation by androstanol suggests CAR may be important in sex hormone homeostasis as well. We hypothesized that sex hormone levels are altered in CAR knockout (CARKO) mice. Serum testosterone and androstenedione levels were down 2.65 and 2.26 fold respectively in CARKO mice, while estradiol and 17-hydroxyprogesterone levels were the same as wildtype mice. CARKO mice have several physiological phenotypes accompanying the loss of androgens, including a significant increase in testes weight (38%), and atrophy of the prostate and seminal vesicles. Histological examination of CARKO testes by H[amp]E-PAS staining show decreased leydig cell islands, and cellular debris in seminiferous tubules suggestive of impaired spermatogenesis, due to loss of androgen signaling. QPCR analysis of transcript levels of detoxification pathways in the liver and testes suggest that a decrease in androgen levels is not due to an increase in detoxification. Cyp3a, Cyp2b, Cyp2c subfamily members, Sult2a1, and Mrp transporters were not induced in CARKO mice. Steroidogenic enzymes were lower in the liver and more importantly in the testes of CARKO mice. Cyp17a1, Cyp19a1, and Srd5a1 were both more than 2 fold lower in the liver, while Cyp17a1 and Srd5a1 were lower in testes of CARKO mice. Interestingly Srd5a1, which converts testosterone to the more potent androgen dihydrotestosterone (DHT), was almost completely undetectable in CARKO mice. We were able to detect CAR expression in the testes of wildtype mice, in the sertoli cell line (TM4), and we are currently determining CAR expression in leydig cells (TM3) which are responsible for testosterone production in the testes. Hypertrophy of the testes and atrophy of prostate/seminal vesicles was measured in CARKO mice as young as 4 weeks old, and in heterozygous mice containing only one allele of CAR. Our data suggests that CAR is important in androgen homeostasis and we would like to determine if environmental estrogens or anti-androgens could have an effect on reproduction and fecundity involving inhibition of CAR.[br][br]Sources of Research Support: RO1DK046546.[br][br]Nothing to Disclose: JPH, SA, DDM 2012-06-24T12:30:00 362 2012-06-24T00:00:00 1899-12-30T12:30:00 2265 147 978 OR11-6 OR35-01 Sunday 804 2012


805 ENDO12L_OR12-1 ORAL SESSION: Novel Regulators of Kisspeptin [amp] GnRH Signaling (11:15 AM-12:45 PM) Anteroventral Periventricular Cocaine- and Amphetamine-Regulated Transcript Neurons as Potential Mediators of Negative Energy Balance-Induced GnRH Inhibition Cadence True, Saurabh Verma, Melissa Kirigiti, Kevin Grove, Susan Smith Oregon Health [amp] Science University, Beaverton, OR Inhibition of gonadotropin-releasing hormone (GnRH) during conditions of negative energy balance is highly conserved in female mammals; however, the hypothalamic circuitry underlying this inhibition is still not fully understood. Cocaine- and amphetamine-regulated transcript (CART) is a potential candidate linking metabolism and reproduction given its known role in body weight regulation and evidence suggesting CART cells in the anteroventral periventricular region (AVPV) send projections to GnRH cell bodies. Given that the effects of CART upon GnRH release remain largely unknown the current study aimed to investigate the potential role of CART in GnRH regulation, specifically during conditions of negative energy balance.[br]Electrophysiological recordings demonstrated that a subpopulation of GnRH neurons is strongly depolarized by CART. This depolarization persisted in the presence of the sodium channel blocker tetrodotoxin, suggesting at least part of CART[apos]s depolarizing effect may be through a direct post-synaptic mechanism. To determine whether a change in stimulatory CART might contribute to GnRH inhibition during negative energy balance, protein levels in the AVPV were investigated in two models of negative energy balance: lactation and caloric restriction. GnRH inhibition arises during lactation due to the high metabolic output associated with milk production. Immunohistochemistry demonstrated a dramatic increase in AVPV CART protein during lactation, whereas preliminary data indicates corresponding AVPV CART mRNA may be decreased during lactation, suggesting a potential inhibition of protein release and accumulation within cell bodies. Previous work has reported a similar inhibitory regulation of AVPV kisspeptin (Kiss1) during lactation, increased protein and decreased mRNA, but immunohistochemistry demonstrated AVPV CART and Kiss1 cells are separate populations. CART fibers were observed to make close appositions to an estimated 55% of AVPV Kiss1 cells indicating a potential additional indirect Kiss1-mediated mechanism for CART regulation of GnRH. Unlike lactation, caloric restriction did not lead to a change in AVPV CART protein levels. The current findings suggest CART may play a direct and indirect role in stimulating GnRH release and that inhibition of AVPV CART cells may contribute to lactation-specific mechanisms of GnRH inhibition. Future work will investigate how AVPV Kiss1 and CART cells may act in concert to regulate GnRH release.[br][br]Sources of Research Support: NIH Grants HD14643, RR00163, HD18185.[br][br]Nothing to Disclose: CT, SV, MK, KG, SS 2012-06-24T11:15:00 361ABDE 2012-06-24T00:00:00 1899-12-30T11:15:00 836 148 979 OR12-1 OR19-01 Sunday 805 2012


806 ENDO12L_OR12-2 ORAL SESSION: Novel Regulators of Kisspeptin [amp] GnRH Signaling (11:15 AM-12:45 PM) Disruption of Glucocorticoid Receptor Signaling in Kisspeptin Neurons Accelerates the Recovery of Reproductive Function in the Post-Traumatic Stress Period Oulu Wang, Anne Lanjuin, Caroline Ho, Juliana Basko, Catherine Dulac, Joseph Majzoub Children[apos]s Hospital Boston, Boston, MA; Harvard Medical School, Boston, MA; Harvard University, Howard Hughes Medical Institute, Cambridge, MA Stressors can generate adaptive stress responses that prepare for the return to homeostasis, but also maladaptive responses, including the inhibition of reproductive function even after cessation of the stressor. How acute stress causes these maladaptive, post-traumatic effects is not well understood. Kisspeptin (KISS1) is required for the activation of the hypothalamic-pituitary-gonadal (HPG) reproductive axis in humans and mice (1-3). We hypothesized that acute stress in adult mice transiently inhibits kisspeptin neurons and the downstream HPG axis, and that restoration of kisspeptin signaling is necessary to reactivate the axis, similar to its role during puberty. This inhibition of kisspeptin expression could be caused by the concomitant stress-induced elevation of glucocorticoids, with the subsequent fall in glucocorticoids allowing the restoration of kisspeptin expression required for the recovery of the reproductive axis after stress.[br]To test this hypothesis, we first examined the response of hypothalamic [italic]Kiss1[/italic] mRNA expression to different stressors in male mice. After restraint, plasma corticosterone was increased, testosterone was decreased, and [italic]Kiss1[/italic] expression was decreased. After food deprivation, corticosterone was increased, testosterone was decreased, and [italic]Kiss1[/italic] expression was decreased. After cold exposure, corticosterone was unchanged, and [italic]Kiss1[/italic] expression was unchanged, even though testosterone was decreased. All changes were significant. The consistent relationship between a rise in corticosterone and fall in [italic]Kiss1[/italic] expression suggested that elevated corticosterone caused stress-induced inhibition of [italic]Kiss1[/italic] expression. To test this directly, we injected mice with a stress dose of corticosterone and observed that testosterone and [italic]Kiss1[/italic] expression were decreased. To evaluate the role of glucocorticoid signaling in kisspeptin neurons, we generated mice lacking glucocorticoid receptors specifically in kisspeptin-containing neurons. In these animals, [italic]Kiss1[/italic] expression was no longer inhibited during restraint stress, even though corticosterone was increased. Both testosterone and copulatory behaviors showed significantly accelerated recovery in the post-traumatic stress period. Blockade of glucocorticoid receptor signaling in kisspeptin neurons during stress accelerates the recovery of reproductive function during the post-traumatic stress period, and this finding may have therapeutic implications in humans with post-traumatic stress disorders.[br][br](1) Seminara et al., 2003. (2) De Roux et al., 2003. (3) Lapatto et al., 2007.[br][br]Sources of Research Support: DoD NDSEG; NSF GRFP; NIH.[br][br]Nothing to Disclose: OW, AL, CH, JB, CD, JM 2012-06-24T11:30:00 361ABDE 2012-06-24T00:00:00 1899-12-30T11:30:00 859 148 980 OR12-2 OR19-01 Sunday 806 2012


807 ENDO12L_OR12-3 ORAL SESSION: Novel Regulators of Kisspeptin [amp] GnRH Signaling (11:15 AM-12:45 PM) A Role for the Histone Demethylase LSD1 in Controlling the Timing of Pubertal Onset John C Gill, Cecilia Kwong, Erin Clark, Rona S Carroll, Yujiang G Shi, Ursula B Kaiser Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA Recent studies indicate that dynamic modification of histone lysine methylation patterns is involved in the activation, de-repression or silencing of gene networks across development. Our goal has been to identify critical factors associated with the upstream regulation of puberty and the initiation of the transcriptional changes in genes required for increasing GnRH neuronal activity within the hypothalamus. We have identified that lysine-specific demethylase 1 (LSD1), an epigenetic chromatin-remodeling enzyme that can regulate gene activity, influences the central regulation of pubertal timing in mice. LSD1 is present within the juvenile hypothalamus and we have tested the contributions of LSD1 to the neuroendocrine events that mark the initiation of pubertal development [italic]in vivo[/italic] and [italic]in vitro[/italic]. Heterozygous LSD1-deficient (LSD1[sup]+/-[/sup]) female mice have a significant advance in the age of vaginal opening (VO), a marker of pubertal onset, by 3 days compared to wild-type (WT) littermates (LSD1[sup]+/-[/sup], 27.5 days; WT 30.5 days; N=6-9; P[lt]0.05), with no difference between the weights of each genotype. The age of first estrus, another marker of puberty, was also significantly advanced (LSD1[sup]+/-[/sup], 34 [plusmn] 1.6 days; WT, 38 [plusmn] 0.7 days; N=6; P[lt]0.05). Monitoring estrous cyclicity after first estrus revealed that the LSD1[sup]+/-[/sup] mice displayed longer periods in estrus than WT counterparts (LSD1[sup]+/-[/sup], 4.3 [plusmn] 0.2 days/week, WT, 3.1 [plusmn] 0.2 days/week; N=3; P[lt]0.05), indicating a specific disruption of the central regulation of reproduction. At the peripubertal age P27, serum FSH was higher in LSD1[sup]+/-[/sup] females compared to WT (FSH, LSD1[sup]+/-[/sup] 15042 [plusmn] 5097 pg/ml, WT 5209 [plusmn] 1178 pg/ml; P[lt]0.05; LH, LSD1[sup]+/-[/sup] 139 [plusmn] 65 pg/ml, WT 45 [plusmn] 20 pg/ml; P=0.20). Central regulators of puberty in the arcuate nucleus were measured by qRT-PCR in prepubertal (P20) LSD1[sup]+/-[/sup] females and demonstrated increased [italic]Tac2[/italic] expression (2.0 [plusmn] 0.3-fold, P[lt]0.05) and a trend towards increased [italic]Kiss1[/italic] expression (3.0 [plusmn] 0.9-fold, P=0.13) relative to WT levels. Further, LSD1 was associated with the [italic]Kiss1[/italic] promoter by hypothalamic tissue ChIP analysis. In a Kiss1-expressing human breast cancer cell line (MDA-231), knockdown of LSD1 expression by lentiviral shRNA transduction increased [italic]Kiss1[/italic] expression 10-fold, suggesting that LSD1 acts to repress this pubertal regulator. These findings establish LSD1 as a key gating mechanism of pubertal activation and establish a new field of investigation involving the epigenetics of reproductive development.[br][br]Nothing to Disclose: JCG, CK, EC, RSC, YGS, UBK 2012-06-24T11:45:00 361ABDE 2012-06-24T00:00:00 1899-12-30T11:45:00 507 148 981 OR12-3 OR19-01 Sunday 807 2012


808 ENDO12L_OR12-4 ORAL SESSION: Novel Regulators of Kisspeptin [amp] GnRH Signaling (11:15 AM-12:45 PM) SMAD2/3-Mediated Signaling in Gonadotropes Is Dispensable for Fertility and FSH Synthesis in Mice Jerome Fortin, Daniel J Bernard McGill University, Montr[eacute]al, Canada Mammals depend on the dimeric follicle-stimulating hormone (FSH), produced by pituitary gonadotropes, for proper reproductive function. The TGF[beta] superfamily members, activins, are critical regulators of FSH synthesis. In its canonical form, activin signaling is mediated downstream of heteromeric type I/type II receptor complexes via the effector proteins, SMADs 2 and 3, which regulate gene transcription. Converging and extensive evidence from experiments in cells lines indicates that activin-stimulated FSH [beta] subunit (FSH[beta]/[italic]Fshb[/italic]) transcription is SMAD2/3-dependent. These observations suggest that SMAD2/3-mediated signaling is required to maintain proper FSH synthesis and reproductive axis activity. Here, we challenge this widely-held view, showing that mice lacking SMAD2/3 in gonadotropes have preserved fertility and FSH production. We generated mice with gonadotrope-specific deletion of SMAD2/3 (hereafter [italic]Smad2/3[/italic]KO) by crossing Gnrhr-IRES-Cre (GRIC) mice with mice carrying [italic]Smad2[/italic] and [italic]Smad3[/italic] conditional alleles. Female [italic]Smad2/3[/italic]KO mice displayed mild disruptions in estrous cyclicity, characterized by prolonged periods of estrus and diminished cycle frequency. However, their ovarian and uterine weights were normal and their fertility was intact, both in terms of litter size and frequency. [italic]Smad2/3[/italic]KO males had slightly reduced testes weight but had normal seminal vesicle weight and were fertile. Unexpectedly, pituitary [italic]Fshb[/italic] expression was unchanged in both male and female [italic]Smad2/3[/italic]KO mice compared to controls. Furthermore, serum FSH levels were normal in [italic]Smad2/3[/italic]KO males, whereas mean diestrus serum FSH was elevated by about 1.7-fold in female [italic]Smad2/3[/italic]KO mice. To verify whether SMAD2/3 are required for activin-regulated expression of [italic]Fshb[/italic] in primary gonadotropes, we prepared cultures of pituitary cells from mice carrying [italic]Smad2/3[/italic] conditional alleles and recombined the genes [italic]ex vivo[/italic] by infecting cells with a Cre-expressing adenovirus. Surprisingly, despite near-complete ablation of SMAD2/3 ([gt]95%), basal [italic]Fshb[/italic] expression was only slightly reduced, and exogenous activin could still stimulate [italic]Fshb[/italic] expression. Together, our data show that, contrary to a long-standing assumption, FSH synthesis and fertility do not require SMAD2/3-mediated signaling in gonadotropes [italic]in vivo[/italic].[br]The authors thank Drs Ulrich Boehm, Jonathan Graff, Martin Matzuk and Michael Weinstein for their generous gift of the mouse lines used in this study.[br][br]Sources of Research Support: This work was supported by CIHR grant MOP-89991 to DJB.[br][br]Nothing to Disclose: JF, DJB 2012-06-24T12:00:00 361ABDE 2012-06-24T00:00:00 1899-12-30T12:00:00 1033 148 982 OR12-4 OR19-01 Sunday 808 2012


809 ENDO12L_OR12-5 ORAL SESSION: Novel Regulators of Kisspeptin [amp] GnRH Signaling (11:15 AM-12:45 PM) Disruption of Endogenous Clocks in GnRH Neurons Impairs Reproduction [italic]In Vivo[/italic] Kristen P Tolson, Karen J Tonsfeldt, Cheri P Goodall, Patrick E Chappell Oregon State University, Corvallis, OR; University of California, San Diego, La Jolla, CA; Oregon Health [amp] Sciences University, Portland, OR Reproduction in mammals depends on episodic, pulsatile secretion of gonadotropin-releasing hormone (GnRH) from the hypothalamus, and ovulation in females requires robust surges of this hormone, which are precisely timed such that ova release corresponds with optimal sexual behavior. While GnRH pulses and surges are well characterized, it is unclear precisely what cellular and molecular mechanisms underlie these different modes of peptide release timing. Previous [italic]in vitro[/italic] work in the immortalized GnRH-secreting GT1-7 cell line revealed expression of endogenous circadian clocks, exhibiting oscillations of [ldquo]core[rdquo] clock transcription factors such as Bmal1 acting in intracellular feedback loops. Roles of the molecular clock in reproduction are supported by recent studies: [italic]Clock/Clock[/italic] mutant females display longer, irregular estrous cycles and diminished preovulatory LH surges, and [italic]Bmal1[/italic] knockout mice are infertile. While these reproductive deficits may be due in part to clock disruption in the SCN and other peripheral oscillators, endogenous circadian cycling in GnRH neurons may also be required for normal reproductive function [italic]in vivo[/italic].[br]In order to determine the reproductive role of endogenous circadian clocks in GnRH neurons, we first examined Bmal1 expression in GnRH neurons. We found that a subpopulation of GnRH neurons expresses BMAL1, and the level of colocalization is sexually dimorphic and affected by cycle stage. We then used GnRH-Cre transgenic mice to conditionally delete [italic]Bmal1[/italic] from GnRH neurons [italic]in vivo[/italic]. We evaluated reproductive parameters and found that GnRH-Cre/[italic]Bmal1[/italic][sup]flox/flox[/sup] mice exhibit subfertility, but normal fecundity. Conditional homozygous deletion of [italic]Bmal1[/italic] from GnRH neurons did not significantly affect estrous cyclicity in adult females, but did cause abnormal LH secretion after ovariectomy (OVX) and 17[beta]-estradiol (E2) priming. Our results suggest that endogenous clocks play an important role in GnRH neurons, regulating the timing of the GnRH/LH surge and influencing reproductive function.[br][br]Sources of Research Support: NIH Grant HD065331 awarded to PEC.[br][br]Nothing to Disclose: KPT, KJT, CPG, PEC 2012-06-24T12:15:00 361ABDE 2012-06-24T00:00:00 1899-12-30T12:15:00 529 148 983 OR12-5 OR19-01 Sunday 809 2012


810 ENDO12L_OR12-6 ORAL SESSION: Novel Regulators of Kisspeptin [amp] GnRH Signaling (11:15 AM-12:45 PM) Kisspeptin Cell-Specific Deletion of ER[alpha] Abrogates Both Stimulatory and Inhibitory Effects of Estradiol-17[beta] (E[sub]2[/sub]) on Kisspeptin Gene Expression in the Female Hypothalamus Sharon L Dubois, Maricedes Acosta-Martinez, Christian Mayer, Mary DeJoseph, Andrew Wolfe, Sally Radovick, Ulrich Boehm, Janice H Urban, Jon E Levine University of Wisconsin, Madison, WI; Stony Brook University Medical Center, Stony Brook, NY; Center for Molecular Neurobiology, Hamburg, Germany; Rosalind Franklin University of Medicine and Science, North Chicago, IL; Johns Hopkins University School of Medicine, Baltimore, MD; University of Wisconsin, Madison, WI Female kisspeptin cell-specific estrogen receptor [alpha] knockout (KERKO) mice exhibit advanced onset of, and incomplete progression through, puberty and remain largely anovulatory in adulthood (1). We hypothesize that this complex pubertal phenotype arises from the loss of ER[alpha]-mediated inhibition of kisspeptin expression in the arcuate nucleus (ARC), leading to early activation of GnRH neurosecretion, and the lack of ER[alpha]-mediated stimulation of kisspeptin expression in the anteroventral periventricular (AVPV) neurons, preventing the pubertal up-regulation of GnRH release. To determine whether ER[alpha] in AVPV and ARC kisspeptin neurons mediate stimulation and inhibition of kisspeptin gene expression, respectively, we examined the effects of E[sub]2[/sub] treatments on kisspeptin mRNA expression in the presence and absence of ER[alpha] in kisspeptin neurons. Adult wild-type (WT) and KERKO female mice were ovariectomized and treated with vehicle (veh) or E[sub]2[/sub]-filled s.c. capsule implants for 1 wk. Kisspeptin mRNA levels in the AVPV and ARC were analyzed by [italic]in situ[/italic] hybridization, emulsion autoradiography, and quantitative image analysis. Kisspeptin mRNA expression in the AVPV was low in veh-treated WT mice and increased by 4-fold in E[sub]2[/sub]-treated WT mice. Kisspeptin mRNA was undetectable in the AVPV of veh-treated KERKO mice and increased in E[sub]2[/sub]-treated animals to the low levels observed in veh-treated WT mice. In contrast to the basal kisspeptin mRNA levels seen in the AVPV, veh-treated WT and KERKO mice expressed kisspeptin mRNA at similar levels in the ARC; however, although E[sub]2[/sub] treatment significantly reduced the amount of kisspeptin mRNA in WT mice, it was without effect in the ARC of KERKO mice. Thus, ER[alpha] in kisspeptin neurons mediates a major part of the stimulatory effects of E[sub]2[/sub] on kisspeptin expression in the AVPV and all of E[sub]2[/sub][apos]s inhibitory effects on kisspeptin gene expression in the ARC. These findings are consistent with the hypothesis that the premature onset and incomplete progression of puberty in KERKO mice results from the abrogation of ER[alpha]-mediated positive and negative effects on kisspeptin expression in AVPV and ARC neurons, respectively. Since diminished kisspeptin expression in the AVPV is a feature of the KERKO phenotype, these data also reinforce the idea that the activation of ER[alpha] in these neurons is an obligatory step in the development of kisspeptin neurons, and in the sequence of neuroendocrine events that comprise pubertal maturation.[br][br](1) Mayer C et al., Proc Natl Acad Sci 2010; 107(52):22693-22698.[br][br]Sources of Research Support: Deutsche Forschungsgemeinschaft Grant DFG BO1743/2; NIH grants K99 HD55446, P01 HD21921, T32 HD07068, P50 HD44405.[br][br]Nothing to Disclose: SLD, MA-M, CM, MD, AW, SR, UB, JHU, JEL 2012-06-24T12:30:00 361ABDE 2012-06-24T00:00:00 1899-12-30T12:30:00 1010 148 984 OR12-6 OR19-01 Sunday 810 2012


811 ENDO12L_OR13-1 ORAL SESSION: Osteoblasts [amp] Bone Biology (11:15 AM-12:45 PM) Postnatal Ablation of Gs[alpha] in Osteoblasts Results in Osteoporosis and Impaired Bone Marrow B-Cell Development Piia Aarnisalo, Gregory Nachtrab, Cristina Panaroni, Partha Sinha, Rhiannon Chubb, Min Chen, Lee S Weinstein, Ernestina Schipani, Henry M Kronenberg, Joy Y Wu Massachusetts General Hospital, Boston, MA; University of Helsinki, Helsinki, Finland; NIDDK, Bethesda, MD; Indiana University, Indianapolis, IN Parathyroid hormone is an important regulator of osteoblast function and is in clinical use for the treatment of osteoporosis in humans. The parathyroid hormone receptor (PPR) is a G protein-coupled receptor that signals via multiple G proteins including Gs[alpha]. In humans with a mutation in PPR leading to constitutive receptor activation and augmented intracellular cAMP, and in transgenic mice expressing such a mutant receptor in osteoblasts (caPPR mice), trabecular bone is dramatically increased. In contrast, we have reported that ablation of Gs[alpha] in the osteoblast lineage during embryonic developments using osterix-driven cre recombinase (Gs[alpha]OsxKO mice) results in fractures at birth. To investigate the contribution of Gs[alpha] to the increase in trabecular bone resulting from activation of the PPR, we crossed Gs[alpha]OsxKO mice to transgenic caPPR mice. However, expression of the caPPR transgene in Gs[alpha]OsxKO mice failed to prevent the fractures, demonstrating that Gs[alpha] is a major downstream mediator of the actions of caPPR in bone. We then questioned whether Gs[alpha] signaling is essential not only in the development of bone, but also in adult bone homeostasis. Since Gs[alpha]OsxKO mice die shortly after birth, they could not be used to investigate the function of Gs[alpha] signaling in the adult skeleton. However, administration of doxycycline during embryogenesis delayed the ablation of Gs[alpha] until the postnatal period, and the resulting mutant mice survived well into adulthood. Postnatal deletion of Gs[alpha] in the osteoblast lineage resulted in osteoporosis with reduced trabecular and cortical bone, as well as increased expression in osteocytes of sclerostin, an inhibitor of canonical Wnt signaling. Fetal ablation of osteoblastic Gs[alpha] demonstrated that Gs[alpha] signaling in osteoblasts is also required for normal bone marrow development of B cell precursors. In postnatal Gs[alpha]OsxKO mice, a similar impairment of B lymphocyte lineage differentiation is found at the pre-pro-B cell to pro-B cell transition. Thus Gs[alpha] signaling in osteoblasts is crucial not only during embryonic development, but also in the postnatal skeleton for normal bone mass and marrow B lymphocyte differentiation. Postnatal conditional ablation of Gs[alpha] in osteoblasts is a valuable model for studying the mechanisms and treatment of osteoporosis.[br][br]Nothing to Disclose: PA, GN, CP, PS, RC, MC, LSW, ES, HMK, JYW 2012-06-24T11:15:00 342ABDE 2012-06-24T00:00:00 1899-12-30T11:15:00 378 149 985 OR13-1 OR04-01 Sunday 811 2012


812 ENDO12L_OR13-2 ORAL SESSION: Osteoblasts [amp] Bone Biology (11:15 AM-12:45 PM) Dissecting the Role of Osteoblast-Derived Nitric Oxide in Bone Remodeling Monica Grover, Sandesh Chakravarthy Sreenath Nagamani, Ayelet Erez, Brendan Lee Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX [bold]Background:[/bold] Bone accrual during adolescence is the main contributor to peak bone mass and is a key determinant of bone health throughout life. Bone remodeling is a balance between bone formation and bone resorption as directed by osteoblast and osteoclast activity, respectively. Nitric oxide (NO) is a potent regulator of bone remodeling via mediating effects of cytokines, estrogen and mechanical strain. NO is synthesized from the conversion of L-arginine to L-citrulline by the enzyme Nitric Oxide Synthase (NOS). Citrulline can be reconverted to arginine by the enzymes argininosuccinate synthase (ASS) and argininosuccinate lyase (ASL). [bold]ASL is the sole mammalian enzyme responsible for endogenous L-arginine production and routing exogenous arginine to NOS for NO synthesis. Deletion of ASL abolishes arginine dependent NO production in the cell [/bold](Erez et al [italic]Nature Medicine[/italic] 2011). Multiple studies have attempted to determine the role(s) of NO in bone remodeling. [italic]However, as of yet, no study has been able to dissect its role in a cell specific manner. [/italic][br][bold]Hypothesis: [/bold]NO produced by osteoblasts stimulates osteoblast proliferation and increases bone mass in homeostasis and under hormonal stress.[br][bold]Research Design and Methods:[/bold] Using Cre-Lox technology, we created an osteoblast specific knockout of [italic]Asl in the mouse model. [/italic]Bilateral ovariectomy was performed at 12 weeks of age to induce estrogen deficiency. Osteoblast activity and bone remodeling was evaluated at 12, 20 and 24 weeks using bone turnover markers, [micro]CT and histomorphometry. [italic]Asl[/italic] cKO female mice were compared to their sham-operated and [italic]Asl[/italic] [italic][sup]Flox/Flox[/sup][/italic] littermates at each time point.[br][bold]Preliminary Results: [/bold]At 12 weeks of age, [italic]Asl[/italic] cKO female mice have significantly lower trabecular bone density (BV/TV), trabecular number (Tb.N.) and higher trabecular separation (Tb.Sp.) in lumbar spine when compared to [italic]Asl[sup]Flox/Flox [/sup][/italic]littermate female controls (p[lt]0.05).[br][bold]Discussion: [/bold]Our preliminary results show that [italic]Asl[/italic] cKO female mice have lower bone mass in lumbar spine at baseline suggesting an inherent role of NO in normal bone mineralization. Evaluation of bone remodeling in the [italic]Asl[/italic] cKO female mice under an estrogen deficient state is ongoing. The translational value of the [italic]Asl[/italic] cKO model is potential development of pharmacologic and genetic manipulation of ASL as an effective regulator of NO metabolism in different NO dysfunctional states, ulimately leading to improved bone mineralization.[br][br]Nothing to Disclose: MG, SCSN, AE, BL 2012-06-24T11:30:00 342ABDE 2012-06-24T00:00:00 1899-12-30T11:30:00 869 149 986 OR13-2 OR04-01 Sunday 812 2012


813 ENDO12L_OR13-3 ORAL SESSION: Osteoblasts [amp] Bone Biology (11:15 AM-12:45 PM) Nitric Oxide Works through Insulin-Like Growth Factor I, in Prevention of Bone Loss with Nitroglycerin Sunil J Wimalawansa UMDNJ- Robert Wood Johnson Medical School, New Brunswick, NJ Introduction: Diagnostic and therapeutic advances have made over the past decade in treatment of osteoporosis. Albeit new therapies are effective, these are expensive and have adverse effects. Therefore, cost-effective therapeutic options are needed. We previously reported that beneficial effects of estrogen on bone is at least in part mediated via nitric oxide (NO)/cGMP pathway, and via IGF-1 (1-3). At appropriate doses, nitroglycerin (NG) as a nitric oxide (NO) donors uncouple osteoblasts and osteoclasts, and have positive effect on bone (3,4).[br]Methods: A 3-year randomized, doubled-blind, controlled clinical trial was conducted to assess the efficacy NG in preventing bone loss in postmenopausal women. Study, Nitroglycerin as an Option, Value in Early Bone Loss (NOVEL) was funded by NIAMS. Women were randomized to receive either nitroglycerin ointment or placebo ointment. All women received calcium and vitamin D supplements. There were no statistically significant differences in the BMD in the treatment vs. calcium and vitamin D arms (5). However, taking compliance into consideration ([sim]75%), average dose actually used by the study participants was sub-optimal; [sim]50% of that was intend to use in the study.[br]Results: Women who had positive BMD response following NG therapy had a significant increase of serum IGF-1 levels, but not in placebo-treated subjects even those who gained BMD. NG-treated subjects with increased BMD had increase IGF-1 levels, 201 [plusmn] 25.6 vs. non-responders, 40.2 [plusmn] 16.9 ng/mL (p[lt]0.001), and the BMD changes was positively correlated with the change of serum IGF-1 levels from the baseline (r = 0.5; p[lt]0.01). Whereas, those who were in the placebo group with increased BMD had no change in serum IGF-1 levels (-2.6 [plusmn] 24.6 vs. 10.8 [plusmn] 13.5 ng/mL, NS; responders vs. non-responders).[br]Conclusions: We previously demonstrated that NO synthase inhibitors such as L-NAME block estrogen effects on bone. Current data suggest that NG, in addition to be a key final common pathways for positive effect of estrogen in bone, may also involved in enhancing the local production of IGF-1, and enhancing bone formation that is observed with nitric oxide therapy (3,6). Nitroglycerin, is one of the key final common pathways for positive effect of estrogen in bone, is also be involved in enhancing local production of IGF-1, thereby increasing bone formation following treatment with the appropriate doses of NO donors.[br][br](1) Wimalawansa SJ et al., Bone, 1997; 21: 275-280. (2) Wimalawansa SJ. JBMR, 2000; 15: 2240-2244. (3) Wimalawansa SJ. Expert Opinion in Pharmacotherapy, 2008; 9: 3025-3044. (4) Jamal et al., JAMA, 2011, 305: 800-807. (5) Wimalawansa SJ et al, JCEM, 2009; 94: 3356-3364. (6) 29. Wimalawansa, S.J. Annals of New York Academy of Sciences, 2010; 1192: 391[ndash]403.[br][br]Sources of Research Support: NIH-NIAMS RO1 grant awarded to SJW.[br][br]Nothing to Disclose: SJW 2012-06-24T11:45:00 342ABDE 2012-06-24T00:00:00 1899-12-30T11:45:00 2386 149 987 OR13-3 OR04-01 Sunday 813 2012


814 ENDO12L_OR13-4 ORAL SESSION: Osteoblasts [amp] Bone Biology (11:15 AM-12:45 PM) High Level of FGF23-S129F Mutant Is Secreted into the Circulation of Hyperphosphatemic Familial Tumoral Calcinosis Patients and Is Degraded by a Serum Metalloproteinase Said M Shawar, Ahmad R Ramadan, Bassam R Ali, Manal A Ali, Anne John Arabian Gulf University, Manama, Bahrain; United Arab Emirates University, Al-Ain, United Arab Emirates FGF23 is essential in the homeostasis of phosphate and vitamin D. FGF23 loss of function mutation(s) cause hyperphosphatemic familial tumoral calcinosis (HFTC). Earlier reports suggested that the amino terminus fragment of FGF23-S129F mutant is retained intracellularly while the carboxyl terminal fragment is secreted into circulation. Herein, we demonstrate that the whole molecule of FGF23-S129F is secreted into the circulation of HFTC patients and it is degraded by a serum metalloproteinase. Five patients clinically diagnosed with HFTC and confirmed by DNA sequencing harboring the c.386 C[gt]T; p.S129F mutation were studied. Using FGF23-specific polyclonal antibodies in ELISA assays, we found that intact FGF23 (iFGF23) was 2-3 folds higher in the patients[apos] plasma compared to healthy controls. However, we could not detect the mutant hormone in the sera of any patient using the same assay. In contrast, the level of the C-terminal fragment was 5-6 folds higher in both plasma and sera of HFTC patients compared to normal controls. Additionally, using an ELISA assay that employs two monoclonal antibodies that recognize a discontinuous epitope in FGF23, we failed to detect the iFGF23 in serum or plasma of the patients. However, cocktail proteinase inhibitors added to the serum vacutainer before blood collection preserved the intact molecule. iFGF23 was very low in plasma collected in heparin tubes, while plasma collected in sodium citrate tubes retained [sim]40% of the concentration found in EDTA tubes. Serine-specific proteinase inhibitors or a cocktail inhibitors lacking EDTA failed to rescue the intact molecule suggesting that a metalloproteinase is responsible for proteolysis in plasma undergoing clotting to form serum. Western blot analysis using FGF23-specific polyclonal antibodies showed that stably transfected HEK293 with S129F or S71G secreted high levels of iFGF23, similar to wild type. In addition; confocal microscopy imaging of transiently expressing HeLa and HEK293 cells showed weak staining of the Golgi complex with some vesicular staining resembling the ER exit sites. Presumably, this intracellular staining represent the in transit species of the newly synthesized hormone. Our results suggest that circulatory FGF23 mutant either unable to form a triad of FGF23-Klotho-FGFR1c or binds with low affinity and incapable of initiating intracellular signaling. Also, our findings have important effect on the potential use of the hormone in therapies in HFTC.[br][br]Nothing to Disclose: SMS, ARR, BRA, MAA, AJ 2012-06-24T12:00:00 342ABDE 2012-06-24T00:00:00 1899-12-30T12:00:00 543 149 988 OR13-4 OR04-01 Sunday 814 2012


815 ENDO12L_OR13-5 ORAL SESSION: Osteoblasts [amp] Bone Biology (11:15 AM-12:45 PM) Pathogenetic Chromosomal Rearrangements in a Large Series of Patients with Pseudohypoparathyroidism Type I Giovanna Mantovani, Francesca Marta Elli, Luisa de Sanctis, Paolo Beck-Peccoz, Anna Spada Fondazione IRCCS Ca[apos] Granda Ospedale Maggiore Policlinico, University of Milan, Milan, Italy; University of Turin, Regina Margherita Children[apos]s Hospital, Turin, Italy Pseudohypoparathyroidism (PHP) type I includes two major subtypes, Ia and Ib. About 70% of Ia patients, characterized by Albright hereditary osteodystrophy and multihormone resistance (PTH/TSH/GHRH/gonadotropins), carry mutations in [italic]GNAS[/italic] exons encoding Gs[alpha]. About 60% of Ib patients, with hormone resistance limited to PTH and TSH, have methylation defects within [italic]GNAS[/italic] locus caused by the disruption of long-range imprinting control elements. In particular, the most consistent defect is the loss of imprinting at the exon A/B differentially methylated region (DMR)[italic]. [/italic]The familial form of the disease (AD-PHP-Ib) is typically associated with an isolated loss of imprinting at exon A/B due to microdeletions disrupting the upstream STX16 gene. Conversely, most sporadic PHP-Ib cases have [italic]GNAS[/italic] imprinting abnormalities that involve multiple DMRs, with no identified underlying genetic lesion. Recently, methylation defects were detected in pts with Ia phenotype, suggesting a molecular overlap between the two forms. Despite advances in the determination of molecular mechanisms underlying PHP, 30-40% of patients lack a molecular diagnosis. Moreover, it is unclear whether apparently sporadic imprinting defects are rather secondary to genetic defects. In order to simultaneously investigate for [italic]GNAS[/italic] and [italic]STX16[/italic] deletions/duplications and for[italic] GNAS[/italic] imprinting status (A/B-AS-NESP-XL DMRs), we performed Methylation Specific Multiplex Ligation-dependent Probe Amplification (MS-MLPA) in 96 patients (Ia=48/Ib=48), all negative for Gs[alpha] mutations. In 15 patients we detected rearrangements at [italic]GNAS[/italic] and/or [italic]STX16[/italic] genes. In particular: 1) of 8 PHP-Ia patients: 2 patients with apparent methylation defects carry a deletion of the entire [italic]GNAS[/italic] locus, 1 had A/B loss of methylation (LoM) caused by [italic]STX16[/italic] deletions, 5 had deletions encompassing Gs[alpha] exon 1; 2) of 7 PHP-Ib patients: 5 had A/B LoM associated with [italic]STX16[/italic] deletions and 2 showed both extensive methylation defects and deletions within the AS region. In conclusion: a) MLPA proved to be a reliable method to detect genetic abnormalities associated with PHP-I, both known and novel to the literature; b) PHP-Ia and Ib may be caused by [italic]GNAS[/italic] submicroscopic structural mutations; c) All PHP-I patients negative for Gs[alpha] gene mutations should be considered for further molecular investigations to optimize genetic counselling. Ongoing studies are aimed to characterize the newly detected deletions and to investigate the function of the involved regions.[br][br]Sources of Research Support: Grant from the Italian Ministry of Health to G.M.[br][br]Nothing to Disclose: GM, FME, LdS, PB-P, AS 2012-06-24T12:15:00 342ABDE 2012-06-24T00:00:00 1899-12-30T12:15:00 1463 149 989 OR13-5 OR04-01 Sunday 815 2012


816 ENDO12L_OR13-6 ORAL SESSION: Osteoblasts [amp] Bone Biology (11:15 AM-12:45 PM) BA058, a Novel hPTHrP Analog, Reverses Bone Loss and Improves Bone Strength in Ovariectomized Rats Gary Hattersley, Julie Downall, Maysoun Shomali, Kyla Gallacher, C Richard Lyttle Radius Health, Cambridge, MA BA058 is a novel synthetic analog of PTHrP (1-34) being developed as a bone anabolic therapy for the treatment of osteoporosis. Daily SC BA058 has demonstrated safety and efficacy in Phase I and II clinical trials and is currently enrolling in a Phase III fracture prevention study. The objective of this study was to determine the effect of BA058 on bone quality in a rat model. Two doses (5 ug/kg and 20 ug/kg) of BA058 were administered by once daily subcutaneous injection for 42 days into ovariectomized, osteopenic rats. Bone anabolic effects were examined at the femur and lumbar spine by measuring BMD using DEXA, by quantitative analysis of bone microstructure with microcomputed tomography and mechanical strength testing, compared to placebo and intact control rats. At the end of the bone depletion period femur BMD was decreased by 9.8% in OVX compared with intact controls. BA058 significantly (p[lt]0.05) increased mean femur BMD over baseline measurements for both the 5 ug/kg (+21%) and the 20 ug/kg (+27%) treatment groups. Similarly, at the lumbar spine, BMD was also significantly increased with 5 ug/kg and 20 ug/kg (+28% and +40%, respectively). In contrast, the mean BMD change from baseline in placebo treated rats was +3.3% for the femur and -1.7% for the lumbar spine. BMD values after BA058 treatment with both doses exceeded those of sham intact controls. Increased BMD was accompanied by restoration of trabecular microstructure, where BA058 significantly increased bone volume fraction (BV/TV), trabecular thickness (Tb.Th) and trabecular number (Tb.N), and conversely significantly decreased trabecular separation (Tb.Sp) in both the metaphyseal region of the distal femur and L4 region of the lumbar spine. BA058 also had effects on cortical bone, with significantly increased femoral cortical BMD and cortical thickness (Ct.Th) observed. Mechanical testing showed that BA058 treatment resulted in a significant increase in mechanical properties including increased maximum load and stiffness for both doses compared to the placebo-treated group. At the lumbar vertebral body, increases were observed in maximum load, energy, ultimate strength and toughness when compared to placebo treatment and increases in maximum load, energy, ultimate strength and toughness compared to the intact sham group. Together these results demonstrate a marked bone anabolic effect of BA058 in osteopenic rats, characterized by BMD increase, improved bone microstructure and bone strength.[br][br]Disclosures: GH: Employee, Radius Health. JD: Employee, Radius Health. MS: Employee, Radius Health. KG: Employee, Radius Health. CRL: Employee, Radius Health. 2012-06-24T12:30:00 342ABDE 2012-06-24T00:00:00 1899-12-30T12:30:00 143 149 990 OR13-6 OR04-01 Sunday 816 2012


817 ENDO12L_OR14-1 ORAL SESSION: Animal [amp] Human Studies into Sex Determination (11:15 AM-12:45 PM) FGFR2IIIc Is Required for Testis Determination but Is Dispensable for Subsequent Testis Cord Differentiation and Spermatogenesis Stefan Bagheri-Fam, Meiyun Yong, Anja Dietrich, Terje Svingen, Peter Koopman, Veraragavan Eswarakumar, Vincent R Harley Prince Henry[apos]s Institute of Medical Research, Melbourne, Australia; Monash University, Melbourne, Australia; The University of Queensland, Brisbane, Australia; Yale University School of Medicine, New Haven, CT Loss of Fibroblast Growth Factor Receptor 2 (FGFR2) in mice leads to XY gonadal sex reversal, indicating that FGFR2 acts as the receptor for FGF9 during testis development. FGFR2 exists in two isoforms, FGFR2IIIb and FGFR2IIIc which differ in their FGF ligand-binding specificity. FGFR2IIIb is a low affinity receptor for FGF9 and is not normally expressed in Sertoli cells, and XY Fgfr2IIIb knockout mice do not show abnormalities in testis development. FGFR2IIIc is a high affinity receptor for FGF9 and is expressed in Sertoli cells. However, Fgfr2IIIc knockout male mice were reportedly fertile suggestive of normal testis development. It was speculated that over-expression of the FGFR2IIIb isoform could compensate for the loss of FGFR2IIIc. Alternatively, Fgfr2IIIc knockout male mice might show testicular defects at fetal stages that resolve postnatally. To investigate these possibilities, we examined gonadal development in XY Fgfr2IIIc knockout mice at embryonic days (E) 13.5 and 15.5. Instead of normal testes, XY Fgfr2IIIcKO mice developed ovotestes or ovaries, phenocopying the gonadal defects of Fgf9KO and Fgfr2KO mice (on a mixed genetic background). The ovarian regions of XY Fgfr2IIIcKO gonads lacked expression of the Sertoli cell marker AMH and expressed the female somatic cell marker FOXL2. XY sex reversal extended to germ cells which expressed SCP3, a marker of meiotic prophase. Subsequent analysis of adult XY Fgfr2IIIc knockout mice revealed that around 20% were phenotypic females. These data suggest that the XY Fgfr2IIIc knockout embryos with ovotestes develop into the reported fertile males, while the XY Fgfr2IIIc knockout embryos with ovaries develop into infertile females.[br]Genes such as Wt1 and Sox9 that are critical for gonadal development or sex determination often have subsequent roles in testicular development and function such as in the maintenance of Sertoli cell identity or in nursing germ cells through spermatogenesis. To examine the role of FGFR2 in Sertoli cells after the sex determination phase, we conditionally inactivated Fgfr2 (both isoforms) in Sertoli cells at E13.5 using the AMH-Cre mouse line. These mice did not show obvious defects in testis cord differentiation and spermatogenesis. Moreover, expression of selected male and female markers was unchanged when compared to control mice. In summary, FGFR2IIIc is required for testis determination, but is dispensable for subsequent testis cord differentiation.[br][br]Nothing to Disclose: SB-F, MY, AD, TS, PK, VE, VRH 2012-06-24T11:15:00 352DEF 2012-06-24T00:00:00 1899-12-30T11:15:00 1389 150 991 OR14-1 OR37-01 Sunday 817 2012


818 ENDO12L_OR14-2 ORAL SESSION: Animal [amp] Human Studies into Sex Determination (11:15 AM-12:45 PM) Granulosa Cell Transdifferentiation Resulting from Altered Expression of FOXL2 and SOX9 in the Ovaries of Ex3[alpha][beta]ERKO Double Knockout Mice April K Binder, Katherine A Burns, Karina F Rodriguez, Kenneth S Korach National Institute of Environmental Health Sciences, Research Triangle Park, NC Normal female reproductive function requires the expression of estrogen receptor [alpha] and [beta] (ER[alpha] and ER[beta]) in specific cells of the ovary. Previous characterization of the [alpha][beta]ERKO double mutant ovary demonstrated that loss of both ERs led to the appearance of seminiferous tubule-like structures with Sertoli-like cells that express Sex determining region Y-box 9 ([italic]Sox9)[/italic]. This original [alpha][beta]ERKO was made on a C57BL/6/129S6 background, and the phenotype was lost when backcrossed to a complete C57BL/6 background. New ER [alpha] and [beta] knockout mouse lines were created by global disruption of exon 3 floxed ER[alpha] and ER[beta] and then bred to create Ex3[alpha][beta]ERKO animals. The Ex3[alpha][beta]ERKO animals have a mixed C57BL/6/129S6 background similar to the previous [alpha][beta]ERKOs. Ovaries from the Ex3[alpha][beta]ERKO animals aged 3.5-12 months exhibit disorganized cellular histology, are hemorrhagic and show increased macrophage infiltration and collagen accumulation in the stroma. Around 6-8 months of age, the ovaries have dismorphogenic regions that appear to be transdifferentiated and seminiferous tubule-like structures are observed to varying degrees in animals aged 6-12 months. The expression of [italic]Sox9 [/italic]and fibroblast growth factor 9 ([italic]Fgf9[/italic]), critical factors for differentiation of the embryonic testis are increased in the Ex3[alpha][beta]ERKO ovaries. Conversely, the expression of R-spondin 1 ([italic]Rspo1)[/italic], a gene thought to drive ovarian development, is also significantly increased in the ovaries of Ex3[alpha][beta]ERKO animals compared to control littermates. The ovaries of these animals are receiving signals reported to have sexually dimorphic expression during development of the gonads, suggesting ER signaling is necessary for maintenance of granulosa cell differentiation. Furthermore, the ovaries have regions with a mixed population of cells expressing granulosa cell specific FOXL2, or testis specific SOX9. The Ex3[alpha][beta]ERKO animals provide a unique model to explore the loss of granulosa cell differentiation and postnatal sex reversal of the ovary, and may provide insight into the genetic differences reported in the programming of genes important for maintenance of proper sexual differentiation.[br][br]Nothing to Disclose: AKB, KAB, KFR, KSK 2012-06-24T11:30:00 352DEF 2012-06-24T00:00:00 1899-12-30T11:30:00 1336 150 992 OR14-2 OR37-01 Sunday 818 2012


819 ENDO12L_OR14-3 ORAL SESSION: Animal [amp] Human Studies into Sex Determination (11:15 AM-12:45 PM) Expression Profiles of Lin28/let-7 System in the Rat Hypothalamus along Postnatal Maturation and after Experimental Manipulations of Puberty Susana Sangiao-Alvarellos, Maria Manfredi-Lozano, Victor M Navarro, Francisco Ruiz-Pino, Miguel Sanchez-Garrido, F Cordido, Leonor Pinilla, G A Dissen, Sergio R Ojeda, Manuel Tena-Sempere University of C[oacute]rdoba, C[oacute]rdoba, Spain; Brigham and Women[apos]s Hospital and Harvard Medical School, Boston, MA; Oregon National Primate Research Center, Beaverton, OR; School of Health Science, University of A Coru[ntilde]a and Instituto de Investigaci[oacute]n Biom[eacute]dica de A Coru[ntilde]a (INIBIC), A Coru[ntilde]a, Spain Lin28a and Lin28b are related RNA-binding proteins that inhibit the maturation of miRNAs of the Let-7 family. Lin28 participates in the control of cellular stemness and early embryonic development. Considerable interest has arisen recently concerning other physiological roles of the Lin28/Let-7 axis. In 2009, GWA studies disclosed the link between variations in/around the LIN28B locus and the age of menarche. In addition, Lin28a over-expression delayed puberty onset in mice. These findings suggest the potential involvement of the Lin28/Let-7 system in puberty onset; yet, this possibility has not been experimentally explored. We report here the expression profiles of Lin28 mRNAs, as well as Let-7a and Let-7b miRNAs, in rat hypothalamus during postnatal maturation and in selected models of altered puberty. In male and female rats, hypothalamic Lin28b mRNA displayed very high expression during the neonatal period, markedly decreased during the infantile-to-juvenile transition and reached minimal levels at puberty; this profile was absent in the cortex, thus suggesting tissue-specificity. Hypothalamic Lin28a mRNA showed similar dynamics, but dropped abruptly, with virtually negligible levels from infantile period onwards. Conversely, let-7a and/or let-7b miRNA levels displayed opposite profiles, with minimal neonatal levels and progressive increases along postnatal maturation. Similar trends of Lin28 and/or let-7 expression were detected in the anterior (including POA) and medial-basal hypothalamic areas. Perturbation of brain sex differentiation by neonatal treatment with estrogen or androgen, altered the expression ratios of Lin28/let-7 at the time of puberty, with enhanced Lin28b mRNA and decreased let-7b miRNA levels. Marginal but significant changes in Lin28b and Let-7 levels were also detected in other models of delayed puberty, linked to postnatal underfeeding, early photoperiod manipulation or chronic sub-nutrition. Altogether, our data document dramatic changes in the expression of the Lin28/let-7 axis in the rat hypothalamus during early postnatal maturation preceding puberty, with a marked decline in Lin28a/Lin28b mRNA levels coupled to significant increases in let-7a/let-7b miRNAs. Different (early) manipulations that disturb puberty were associated to perturbation of the above profiles, suggesting the potential involvement of developmental changes in Lin28/let-7 expression at the hypothalamus in the central mechanisms leading to puberty onset.[br][br]Sources of Research Support: BFU2011-25021 (Ministerio de Economia y Competitividad), NSF grant IOS1121691 (SRO).[br][br]Nothing to Disclose: SS-A, MM-L, VMN, FR-P, MS-G, FC, LP, GAD, SRO, MT-S 2012-06-24T11:45:00 352DEF 2012-06-24T00:00:00 1899-12-30T11:45:00 2275 150 993 OR14-3 OR37-01 Sunday 819 2012


820 ENDO12L_OR14-4 ORAL SESSION: Animal [amp] Human Studies into Sex Determination (11:15 AM-12:45 PM) Targeted Genetic Analysis in 46,XY Disorders of Sex Development Based on Phenotype Ranna El-Khairi, Trevor Ian Bunch, Ieuan Arwel Hughes University of Cambridge, Addenbrooke[apos]s Hospital, Cambridge, UK 46,XY Disorders of Sex Development (DSD) encompass a range of causes with variable phenotypes. The majority can be divided into disorders of testis development, androgen biosynthesis or androgen action. It is necessary to target genetic studies to enhance the chance of diagnosis in 46,XY DSD. We analysed a large DSD database to identify clinical and biochemical markers to guide genetic studies, focusing on 5 conditions: 5 alpha-reductase type 2 deficiency (5ARD); 17 beta-hydroxysteroid dehydrogenase type 3 (17BHSD) deficiency; complete and partial androgen insensitivity syndrome (CAIS and PAIS); and gonadal dysgenesis caused by SRY and SF1 mutations. A total of 348 46,XY patients with confirmed mutations in [italic]SRD5A2[/italic], [italic]HSD17B3[/italic], [italic]AR[/italic], [italic]SF1[/italic] and [italic]SRY[/italic] genes were included in the study. Phenotype at presentation, External Masculinization Score (EMS), birth weight, family history, LH, FSH, testosterone (T), dihydrotestosterone (DHT), [Delta]4-androstenedione ([Delta]4), T/DHT, T/[Delta]4 ratios, urinary steroid profiles and pelvic imaging were evaluated. We identified 36 patients (24 families) with [italic]SRD5A2[/italic] mutations (10.3%), 26 patients (22 families) with [italic]HSD17B3[/italic] mutations (7.4%), 271 patients with [italic]AR[/italic] mutations (77.8%) (CAIS, n=197, PAIS, n=65) and 15 patients with gonadal dysgenesis ([italic]SRY[/italic] mutations, n=9; [italic]SF1[/italic] mutations n=6). Normal birth weight for gestational age was positively associated with presence of a mutation. Birth weight SDS was between -2 and +2 in the majority of patients in all 5 conditions. EMS was lowest in patients with CAIS, [italic]SRY[/italic] mutations and highest in patients with PAIS. Almost 100% of patients with CAIS, [italic]17BHSD[/italic] deficiency and [italic]SRY [/italic]and [italic]SF1[/italic] mutations were raised female. Urinary steroid analysis was specific for [italic]SRD5A2[/italic] mutations, T/[Delta]4 ratio [lt]0.7 specific for [italic]HSD17B3[/italic] mutations but a T/DHT ratio [gt]10 was less specific for [italic]SRD5A2[/italic] mutations. Mullerian structures were only found in gonadal dysgenesis with [italic]SRY[/italic] mutations. In our series, all patients with [italic]SF1[/italic] mutations presented in the neonatal period with ambiguous genitalia (n=6, mean EMS 3.3 +/-2.3). None had adrenal insufficiency. Establishing the cause in 46,XY DSD remains a challenge. However, judicious use of clinical and biochemical markers can guide genetic testing with improved diagnosis. This has significant impact on sex assignment and long-term physical, psychological and social outcomes.[br][br]Nothing to Disclose: RE-K, TIB, IAH 2012-06-24T12:00:00 352DEF 2012-06-24T00:00:00 1899-12-30T12:00:00 879 150 994 OR14-4 OR37-01 Sunday 820 2012


821 ENDO12L_OR14-5 ORAL SESSION: Animal [amp] Human Studies into Sex Determination (11:15 AM-12:45 PM) A Novel R-SPONDIN1 ([italic]RSPO1[/italic]) Homozygous Mutation: Variable Phenotype in a Large Consanguineous Brazilian Family with Syndromic [italic]SRY[/italic]-Negative 46,XX Testicular Disorder of Sexual Development (DSD) and Palmoplantar Hyperkeratosis (PPK) Rosana Barbosa Silva, Mirian Yumie Nishi, Sorahia Domenice, Luciane Carneiro Carvalho, Berenice Bilharinho Mendonca, Elaine Maria Frade Costa HC-FMUSP, S[atilde]o Paulo, Brazil [bold]Background[/bold] [italic]RSPO1[/italic] is involved in the [italic]WNT4[/italic] regulation and the [beta]-catenin activation, both important to sexual determination and ovarian differentiation. In addition, it participates in the keratinocytes growth and developmental regulation. To date, two [italic]RSPO1[/italic] mutations have been described in patients with [italic]SRY[/italic]-negative 46,XX Testicular DSD associated with PPK and squamous cell carcinoma (1). [bold]Objective[/bold] To analyze [italic]RSPO1[/italic] in a large consanguineous Brazilian family with four PPK-affected [italic]SRY[/italic]-negative 46,XX Testicular males with ambiguous genitalia. [bold]Patients and Methods[/bold] The index case, a 26 yrs-old man with ambiguous genitalia and PPK, was referred to us for endocrine evaluation. Pelvic ultrasound displayed epididymis, seminal vesicules, prostate and absent M[uuml]llerian structures. Gonadal biopsy showed atrophic testicular parenchyma with imature seminiferous tubules, normal Sertoli cells and no spermatogenesis. The family history was remarkable for several PPK-affected individuals that were born of consanguineous mating. [italic]RSPO1[/italic] coding regions were sequenced and [italic]WNT4[/italic] copy number variation analyzed by MLPA. [bold]Results[/bold] We identified the new homozygous nucleotide change NM_001038633.2:c.1093G[gt]A (exon 6) which led to amino acid substitution (p.Cys102Tyr). The C102Y change was absent in 150 normal controls and segregates as a recessive trait in sixty-one family members. All PPK-affected individuals were homozygous for the mutant gene: four [italic]SRY[/italic]-negative 46,XX males (the propositous, 2 uncles and 1 cousin), four 46,XY males (1 brother, 2 uncles and 1 cousin) and unexpectedly a 46,XX fertile female (cousin). The 46,XY and the 46,XX male affected cousins have bilateral corneal opacities and iris malformations and one 46,XX male uncle had a skin tumor. The [italic]WNT4[/italic] copy number variation analysis was normal. [bold]Discussion[/bold] The C102Y change is located in the second highly conserved furin-like domain, which is essential for Wnt signaling pathway amplification and very probably affects protein function. To explain the absence of sex reversal in the 46,XX affected woman we hypothesize that an associated duplication of [italic]WNT4[/italic] balanced de [italic]RSPO1[/italic] mutation. However the [italic]WNT4[/italic] dosage was normal. [bold]Conclusion[/bold] We described a novel [italic]RSPO1[/italic] mutation (c.1093G[gt]A) associated with 46,XX Testicular DSD and PPK in a highly informative consanguineous family. The absence of sex reversal in one 46,XX member remains undetermined. The PPK-affected woman array-CGH analysis probably would elucidate the distinct sex phenotype.[br][br](1) Parma P et al., Nat Genet 2006; 38:1304.[br][br]Nothing to Disclose: RBS, MYN, SD, LCC, BBM, EMFC 2012-06-24T12:15:00 352DEF 2012-06-24T00:00:00 1899-12-30T12:15:00 784 150 995 OR14-5 OR37-01 Sunday 821 2012


822 ENDO12L_OR14-6 ORAL SESSION: Animal [amp] Human Studies into Sex Determination (11:15 AM-12:45 PM) Clinical Management of Youth with Gender Dysphoria at BC Children[apos]s Hospital: Over 10 Years[apos] Experience Karine Khatchadourian, Daniel L Metzger University of British Columbia, Vancouver, Canada Background/Aim: To describe patient characteristics at presentation, treatment and response to treatment in youth with gender dysphoria.[br]Methods: A retrospective chart review of 81 youth with a diagnosis of gender dysphoria seen from 1998[ndash]2011.[br]Results: Of the 81 patients, 42/81 (52%) identified as female-to-male (FTM), 37/81 (46%) as male-to-female (MTF), and 2/81 (2.5%) natal males were undecided. Median age of presentation for FTM youth was 17.0 years (range 11.4[ndash]19.8 years), and median age of presentation for MTF youth was 16.6 years (range 12.3[ndash]22.5 years). GnRHa treatment was prescribed in 27/81 (33%) patients, and of these, 44.4% received their first dose on their first visit. One FTM patient developed sterile abscesses with Lupron Depot[reg]; he was switched to Decapeptyl[reg] CR and tolerated this well. Another FTM patient stopped Lupron Depot[reg] 2 months after initiating treatment due to estrogen-withdrawal symptoms. Cross-sex hormones were prescribed in 61/81 patients (37/42 FTM vs. 24/37 MTF, p[lt]0.02 by [chi]2). Median age of initiation of testosterone injections in FTM patients was 17.8 years (range 13.7[ndash]23.7 years). Median age of initiation of estrogen therapy in MTF patients was 17.9 (range 13.3[ndash]22.3 years). Three patients stopped cross-sex hormones temporarily for reasons not related to transitioning, but due to psychiatric co-morbidities (2 FTM) or testosterone effects causing androgenic alopecia (1 FTM). No severe complications were noted in patients treated with testosterone or estrogen.[br]Conclusion: Treatment with GnRHa and/or cross-sex hormones in collaboration with transgender-competent mental health care professionals is an appropriate intervention in carefully selected youth with gender dysphoria. Long-term follow-up studies are needed to determine the safety of these treatments in this age group.[br][br]Nothing to Disclose: KK, DLM 2012-06-24T12:30:00 352DEF 2012-06-24T00:00:00 1899-12-30T12:30:00 1166 150 996 OR14-6 OR37-01 Sunday 822 2012


823 ENDO12L_OR15-1 ORAL SESSION: Novel Front Door [amp] Back Door Pathways in Steroid Synthesis [amp] Metabolism (11:15 AM-12:45 PM) 5[alpha]-Reductase and 17,20-Lyase Regulate the Activity of the Alternative [apos]Backdoor[apos] Pathway in Patients with 21-Hydroxylase Deficiency Clemens Kamrath, Michaela F Hartmann, Thomas Remer, Stefan A Wudy Justus-Liebig University, Giessen, Germany; University of Bonn, Dortmund, Germany Background: 17-hydroxyprogesterone (17-OHP) can be metabolized to dihydrotestosterone (DHT) via an alternative [apos]backdoor[apos] route that bypasses the conventional intermediates androstenedione and testosterone. In this [apos]backdoor[apos] pathway, 17-OHP is converted to 5[alpha]-pregnane-3[alpha],17[alpha]-diol-20-one (5[alpha]-pdiol) by sequential 5[alpha]-reductase and 3[alpha]-hydroxysteroid dehydrogenase activities. 5[alpha]-pdiol is an excellent substrate for the 17,20-lyase activity of CYP17A1 to produce androsterone.[br]Objective: The aim of this study was to analyze our hypothesis, that the activities of the 5[alpha]-reductase and the 17,20 lyase regulate the alternative backdoor pathway activity in patients with 21-hydroxylase deficiency (21-OHD).[br]Methods: We compared urinary steroid hormone profiles determined by gas chromatography-mass spectrometry of 142 untreated 21-OHD patients (age range: 1 day to 25.4 years; 51 males) with 138 control subjects. We assessed pathway and enzyme activities using product to substrate ratios.[br]Results: The relative activities of the alternative backdoor pathway and 5[alpha]-reductase correlated significantly (rs = 0.67; p[lt]0.0001). Neonates with 21-OHD demonstrated a moderate activity of the 5[alpha]-reductase leading to moderate 5[alpha]-pdiol generation. Due to a very high activity of the 17,20 lyase, 5[alpha]-pdiol is converted with high activity to androsterone. During infancy, the activity of the 5[alpha]-reductase is very high, leading to a high activity of the alternative backdoor pathway until the generation of 5[alpha]-pdiol. Only a moderate androsterone generation is the result of low 17,20 lyase activity in this age group. Children [gt] 1 year of age show a low 5[alpha]-reductase and a moderate 17,20 lyase activity of the alternative backdoor pathway, leading to a low androsterone generation in the backdoor pathway.[br]Discussion: The activity of the backdoor pathway is regulated by the 5[alpha]-reductase and 17,20 lyase activities. The 5[alpha]-reductase is a gatekeeper to the alternative backdoor pathway and regulates its activity until the generation of 5[alpha]-pdiol. Thereafter, the generation of androsterone is regulated by the 17,20 lyase activity. Neonates with 21-OHD demonstrated a high 17,20 lyase activity of the alternative backdoor pathway. During childhood, the activity of the 17,20 lyase of the classic pathway increased more than that of the alternative backdoor pathway, probably influenced by cytochrome b5, leading to an decreasing role of the alternative backdoor pathway for androgenic steroid production.[br][br]Nothing to Disclose: CK, MFH, TR, SAW 2012-06-24T11:15:00 360 2012-06-24T00:00:00 1899-12-30T11:15:00 1410 151 997 OR15-1 OR25-01 Sunday 823 2012


824 ENDO12L_OR15-2 ORAL SESSION: Novel Front Door [amp] Back Door Pathways in Steroid Synthesis [amp] Metabolism (11:15 AM-12:45 PM) [italic]In Vivo[/italic] Evidence for Novel Pathway of Vitamin D3 Metabolism Initiated by P450scc Andrzej T Slominski, Tae-Kang Kim, Haleem Shehabi, Igor Semak, Edith Tang, Minh Nguyen, Heather Benson, Elena Korik, Zorica Janjetovic, Jianjun Chen, Charles Yates, Arnold Postlethwaite, We Li, Robert C Tuckey University of Tennessee Heath Science Center, Memphis, TN; Curtin University of Perth, Perth, Australia; Belarusian State University, Minsk, Belarus; University of Western Australia, Crawley, Australia; University of Tennessee Heath Science Center, Memphis, TN; University of Tennessee Heath Science Center, Memphis, TN Following discovery of in vitro P450scc hydroxylation of vitamin D3 generating 20(OH)D3, 20,23(OH)[sub]2[/sub]D3 and 17,20,23(OH)[sub]3[/sub]D3 as the main products (1-3), which can potentially serve as substrates for CYP27B1, we tested whether this pathway operated in vivo through investigation of a metabolism of vitamin D3 in human placenta, adrenal glands and human and pig epidermal keratinocytes. High performance liquid chromatography (HPLC) and LC-mass spectrometry (LC/MS) were used to identify P50scc derived products with retention times (RT) corresponding to the synthetic standards. In human placentas we identified 20(OH)D3, 22(OH)D3, 25(OH)D3, 20,23(OH)[sub]2[/sub]D3, 1,25(OH)2D3, 17,20,23(OH)[sub]3[/sub]D3, 1,20(OH)2D3 and 1,20,23(OH)[sub]3[/sub]D3. The relative rate of transformation, defined as % of the substrate conversion, was 6.57% for 20(OH)D3 and [lt]1% for other compounds. The selectivity for CYP11A1-dependent metabolism of vitamin D3 and preferential accumulation of 20(OH)D3 products was further confirmed by assays on isolated mitochondria from placentas, and the inhibitory effects of 22R-hydroxycholesterol and DL-aminoglutethimide on the production of 20(OH)D3. Involvement of CYP27B1 in metabolism of P450scc generated products was documented by 20(OH)D3 transformation to 1,20(OH)D3 by placental mitochondria. However, 1,20(OH)D3 can also be produced by action of P450scc on 1(OH)D3. Experiments on rat and bovine adrenal glands showed similar pattern with predominant production of 20(OH)D3 (higher than 25(OH)D3) and detection of 20,23(OH)2D3. In keratinocytes levels of detected 20(OH)D3 and 22(OH)D3 were similar, being slightly higher than of 25(OH)D3. Product with a mass and RT corresponding to 20(OH)D3 was also detected in human serum. Thus, we document for the first time a P450scc-dependent in vivo metabolism of vitamin D3. This metabolism is cell type and organ specific as demonstrated by different ratios of the detected products in placenta, adrenal gland and skin keratinocytes, and is modified by CYP27B1. The potential significance of the pathway is marked by our previous demonstration that these compounds are biologically active, while having low calcemic effects (4, 5).[br][br](1) Guryev et al; Proc Natl Acad Sci U S A 2003; 100: 14754. (2) Slominski et al; FEBS J 2005; 272: 4080. (3) Tuckey et al; FEBS J 2008; 275: 2585. (4) Janjetovic et al; J Cell Physiol 2010; 223: 36. (5) Slominski et al; PLoS One 2010; 5: e9907.[br][br]Sources of Research Support: This work was supported by NIH [Grant R01AR052190] to AS, by the University of Western Australia and by the College of Pharmacy at the University of Tennessee Health Science Center.[br][br]Nothing to Disclose: ATS, T-KK, HS, IS, ET, MN, HB, EK, ZJ, JC, CY, AP, WL, RCT 2012-06-24T11:30:00 360 2012-06-24T00:00:00 1899-12-30T11:30:00 1984 151 998 OR15-2 OR25-01 Sunday 824 2012


825 ENDO12L_OR15-3 ORAL SESSION: Novel Front Door [amp] Back Door Pathways in Steroid Synthesis [amp] Metabolism (11:15 AM-12:45 PM) Transient Kinetic Analysis of the Human Steroid 5[beta]-Reductase (AKR1D1) and the Functional Basis of Bile Acid Deficiency Caused by the P133R Mutation in AKR1D1 Yi Jin, Mo Chen, Trevor M Penning Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA Human steroid 5[beta]-reductase (AKR1D1) catalyzes the stereospecific reduction of the double bonds in [Delta][sup]4[/sup]-3-ketosteroids, a unique reaction that introduces a 90[ordm] bend at the A/B ring fusion to yield a 5[beta]-steroid. AKR1D1 is the only enzyme capable of 5[beta]-reduction in humans and is implicated in the metabolism of all classes of steroid hormones except estrogens. In addition, AKR1D1 also plays a critical role in bile-acid biosynthesis to generate the 5[beta]- configuration in primary bile acids, a structural feature essential for the emulsification of fats and fat soluble vitamins. Genetic defects in [italic]AKR1D1[/italic] (e.g., P133R) have been shown to cause bile acid deficiency, a fatal condition for neonates without bile-acid supplementation. Previously, we found that recombinant AKR1D1 displayed a 20-fold variation in [italic]k[/italic][sub]cat[/sub] values for C18-C27 [Delta][sup]4[/sup]-3-ketosteroids. We investigated the kinetic basis of the large variation in [italic]k[/italic][sub]cat[/sub] values and of the effect of the disease-causing P133R mutation on enzyme function. In stopped-flow multiple turnover experiments, partial or full burst phases were observed at saturating substrate concentrations for fast substrates ([italic]k[/italic][sub]cat[/sub] [gt] 8 min[sup]-1[/sup]), e.g., aldosterone, cortisone, and testosterone, indicating that the chemical events are faster ([italic]k[/italic][sub]burst[/sub] [gt] 30 min[sup]-1[/sup]) than the product release steps which are rate limiting for these substrates. In contrast, burst-phase kinetics were absent in multiple turnover experiments using slow substrates ([italic]k[/italic][sub]cat[/sub] [lt] 2 min[sup]-1[/sup]), e.g., 4-cholsten-7[alpha]-ol-3-one (a bile-acid precursor) and cholestenone, suggesting that the overall rate of reduction of these substrates are limited by the chemical step and not by product release steps. Thus the rate determining steps of AKR1D1 catalysis are steroid substrate dependent. The transient kinetic behavior of the P133R mutant was then compared to wild type AKR1D1. With the representative fast substrate cortisone ([italic]k[/italic][sub]cat[/sub] of 0.7 min[sup]-1[/sup] with P133R), burst-phase kinetics were no long observed, and data revealed a 2-orders of magnitude reduction in the rate of the chemical step for this substrate. With the representative slow substrate 4-cholesten-7[alpha]-ol-3-one ([italic]k[/italic][sub]cat[/sub] of [sim]0.2 min[sup]-1[/sup] with P133R), the burst phase was absent as with the wild type enzyme and a [sim]10-fold reduction in the reaction rate was observed. Therefore, the compromised ability of AKR1D1 to carry out the chemical reaction is the molecular defect causing bile-acid deficiency in patients with P133R mutation.[br][br]Sources of Research Support: (Supported by NIH grants R01-DK47015 and P30-ES013508 awarded to T.M.P.).[br][br]Nothing to Disclose: YJ, MC, TMP 2012-06-24T11:45:00 360 2012-06-24T00:00:00 1899-12-30T11:45:00 457 151 999 OR15-3 OR25-01 Sunday 825 2012


826 ENDO12L_OR15-4 ORAL SESSION: Novel Front Door [amp] Back Door Pathways in Steroid Synthesis [amp] Metabolism (11:15 AM-12:45 PM) Dysregulation of the Local Skin Glucocorticoid Pathway in Psoriasis May Provide Novel Therapeutic Opportunities Rosalind F Hannen, Michael Wright, Anthony E Michael, Abha Gulati, Catherine Harwood, Jacky M Burrin, Michael P Philpott Barts and The London, Queen Mary[apos]s School of Medicine and Dentistry, London, UK; Addenbrookes Hospital, Cambridge, UK; St George[apos]s, University of London, London, UK; Royal London Hospital, Whitechapel, UK; Barts and The London, Queen Mary[apos]s School of Medicine and Dentistry, London, UK Psoriasis is a common hyperproliferative inflammatory skin condition affecting up to 3% of people worldwide. The cause of psoriasis is unknown although it appears to be triggered by environmental and genetic (particularly immune) factors. Glucocorticoids (GC) induce powerful immune-modulatory effects and are therapeutically used to manage psoriasis since GC reduce inflammation, decrease keratinocyte skin cell proliferation and promote keratinocyte differentiation (1). We previously demonstrated [italic] de novo [/italic] cortisol synthesis in normal primary human keratinocytes (NHK) (2). Interestingly, knockout of the GC receptor (GR) in mouse skin induces hyperproliferation of keratinocytes in the absence of inflammatory stimulus (3). Thus the disruption of the GC pathway could be an important pathogenic factor in psoriasis and may provide a novel target for therapeutic intervention.[br]Here we show that the GC pathway is downregulated in human psoriatic skin. Expression of steroidogenic acute regulatory protein was decreased in both lesional and uninvolved psoriatic skin. Thin layer chromatography showed reduced pregnenolone to cortisol metabolism in primary psoriatic keratinocytes (PPK) over 24h (38.8[plusmn]6.1% NHK vs. 8.7[plusmn]0.4% uninvolved and 8.6[plusmn]0.2% lesion PPK, P[le]0.01). In addition, steroid sulphate levels in PPK increased (10.5[plusmn]1.7% NHK vs. 19.5[plusmn]1.9% uninvolved and 22.8[plusmn]3.0% lesion PPK, P[le]0.05), suggesting greater steroid clearance from the system. Cortisol release from whole skin mounts was undetectable by ELISA in lesion and uninvolved psoriatic skin but was measured at 0.40[plusmn]0.14ng/ml in normal skin over 24h. Finally, total GR expression was not detected in 18 and decreased in 2 of the 20 psoriasis lesion biopsies analysed. In contrast, both total GR and phosphorylated (active) GR were strongly detected by immunofluorescence histochemisty in normal skin and patient matched uninvolved psoriatic skin.[br]We can now conclude that the local GC pathway from synthesis to GR expression is attenuated in psoriatic skin. Decreased GC synthesis in lesion and uninvolved skin would provide an environment favorable for promotion of inflammation that is well defined in psoriasis. Loss of GR expression in psoriatic lesions may directly induce keratinocyte hyperproliferation. Therefore dysregulation of localised keratinocyte GC pathway may be implicated in the pathogenesis of psoriasis and represent a novel therapeutic target.[br][br](1) Stojadinovic et al., JBC 2007; 282(6):4021. (2) Hannen et al., BBRC 2011; 404:62. (3) Bayo et al., Endocrinology 2008; 149(3):1377.[br][br]Sources of Research Support: Giuliani S.p.A.[br][br]Nothing to Disclose: RFH, MW, AEM, AG, CH, JMB, MPP 2012-06-24T12:00:00 360 2012-06-24T00:00:00 1899-12-30T12:00:00 730 151 1000 OR15-4 OR25-01 Sunday 826 2012


827 ENDO12L_OR15-5 ORAL SESSION: Novel Front Door [amp] Back Door Pathways in Steroid Synthesis [amp] Metabolism (11:15 AM-12:45 PM) Induction of Human CYP19A1 Gene by Weight Gain in the Adipose Tissue Evidenced by hARO-Luc Reporter Mice Emrah Yatkin, Niina Saarinen, Lauri Polari, Anu Salminen, Leena Strauss, Matti Poutanen, Sari Makela University of Turku, Turku, Finland; University of Turku, Turku, Finland Obesity is associated with elevated estrogen concentrations in men and postmenopausal women, thus, contributing to the development of hormone dependent diseases, most notably to postmenopausal breast cancer. Low-grade inflammation and altered adipokine and cytokine milieu, present in the obese subjects, are suggested to upregulate aromatase expression in the adipose tissue, resulting in elevated local estrogen concentration in, e.g., breast stroma. So far, experimental studies addressing the regulation of human aromatase gene expression in the adipose tissue in vivo have been hampered by the lack of appropriate rodent models, due to significant differences in the regulatory regions of the human and rodent aromatase (CYP19A1) genes. In human aromatase gene, 10 tissue-specific promoters have been reported, and at least 3 of these have been detected in adipose tissue, while mouse aromatase gene is less complex and mainly expressed in gonads and brain. To study the tissue-specific regulation of human aromatase gene in vivo, we have generated a transgenic reporter mouse model in FVB/N background (hARO-Luc mouse) with a construct composing of a 100-kbp-long 5[apos]-region of the human CYP19A1 gene attached to a luciferase reporter gene. Weight gain and increased body adiposity in young adult hARO-Luc mice, induced by high fat diet, increased luciferase activity in the mammary fat pad (including gland parenchyma) in ovariectomized and intact females, as well as in the inguinal subcutaneous fat in males. Similar trend was observed in gonadal fat, while no response was seen in other fat depots (mesenteric, retroperitoneal). Furthermore, obese hARO-Luc mice presented altered serum adipokine and cytokine profiles as well as increased number of crown like structures in the adipose tissue, similar to obese humans and indicating low-grade inflammatory state. We, furthermore, isolated bone marrow derived mesenchymal stromal cells from the hARO-Luc mice, and showed that dexamethasone and PGE2, known to drive CYP19A1 expression in human adipose tissue by promoters I.4 and I.3/PII, stimulated luciferase activity in the reporter mice ex vivo. In conclusion, we have demonstrated that hARO-Luc mouse model is a powerful tool to study the tissue-specific regulation of the human aromatase gene in vivo, and our data suggest that weight gain, associated with low-grade inflammation, stimulates human CYP19A1 expression in mammary and subcutaneous adipose tissue.[br][br]Sources of Research Support: Academy of Finland; the Finnish Funding Agency for Technology and Innovation; European Commission.[br][br]Nothing to Disclose: EY, NS, LP, AS, LS, MP, SM 2012-06-24T12:15:00 360 2012-06-24T00:00:00 1899-12-30T12:15:00 1050 151 1001 OR15-5 OR25-01 Sunday 827 2012


828 ENDO12L_OR15-6 ORAL SESSION: Novel Front Door [amp] Back Door Pathways in Steroid Synthesis [amp] Metabolism (11:15 AM-12:45 PM) The Role of Sonic Hedgehog in Adrenal Steroidogenesis Marta M Swierczynska, Valeria Lamounier-Zepter, Stefan R Bornstein, Suzanne Eaton Universit[auml]tsklinikum Carl Gustav Carus an der Technischen Universit[auml]t Dresden, Dresden, Germany; Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany Hedgehog (Hh) proteins are secreted signaling molecules that control many aspects of human embryogenesis. Their expression continues postnatally, as they are involved in many physiological processes. Disruptions in the Hh signaling pathway not only contribute to multiple congenital diseases but also seem to be associated with the development of many cancer types. In rodents, Sonic hedgehog (Shh) is essential for proper adrenal gland organogenesis, and its expression continues in the adult adrenal cortex. However, the function of Shh during adulthood is not well understood.[br]Due to double lipid modifications, Hh proteins have high affinity towards cellular membranes, yet, are secreted and transported over many cell diameters. In Drosophila melanogaster Hh proteins associate with lipoprotein particles, which is crucial for inducing long-range target gene expression. However, lipoproteins repress the Hh signaling pathway in a ligand-independent manner. Here, we show similar functions for lipoproteins in human Shh signaling. We further demonstrate that a single cell type can secrete Shh in two distinct forms, as sterol modified, lipoprotein-associated Shh or as non-sterol modified Shh monomers. We show, that association of Shh with lipoproteins alleviates their repressive effect on the Hh signaling pathway. Two distinct forms of secreted Shh exert complimentary activities in in vitro signaling assay, suggesting that they affect the pathway in different ways.[br]As lipoproteins are known to be important modulators of adrenal metabolism, we investigated whether Shh may influence steroidogenesis in humans, either directly or by affecting lipoprotein utilization. We confirmed the expression of Shh and all major Hh signaling pathway components in human adult adrenal glands and the adrenal carcinoma cell line NCI-H295R. We further observed that both lipoprotein-associated and lipoprotein-free forms of Shh differentially increase the secretion of steroid hormones by NCI-H295R cells. Moreover, Shh enhances the stimulatory effect of lipoproteins on adrenal steroidogenesis. Interestingly, the stimulatory effect is restricted to aldosterone and cortisol secretion; no effect of Shh on DHEA secretion is observed.[br]Taken together, our data suggest that additional versatility in Hh signaling can by provided by regulating its mode of release. Furthermore, Shh may be involved in the pathophysiology of adrenal gland during adult life by influencing steroid hormone production.[br][br]Sources of Research Support: DFG, KFO 252, Microenvironment of the Adrenal in Health and Disease.[br][br]Nothing to Disclose: MMS, VL-Z, SRB, SE 2012-06-24T12:30:00 360 2012-06-24T00:00:00 1899-12-30T12:30:00 805 151 1002 OR15-6 OR25-01 Sunday 828 2012


829 ENDO12L_OR16-2 ORAL SESSION: Regulation of Adrenal Cortical Function (11:15 AM-12:45 PM) Involvement of Transducer of Regulated CREB Activity (TORC) in ACTH-Induced Regulation of Steroidogenic Acute Regulatory Protein (StAR) Transcription in Adrenocortical Cell Line Y1-A70 Lorna IF Smith, Mark Olah, Ying Liu, Greti Aguilera National Institute of Child Health and Human Development, Bethesda, MD; University of Bristol, Bristol, UK The stimulatory effect of adrenocorticotropic hormone (ACTH) on glucocorticoid secretion from the adrenal zona fasciculata involves transcriptional activation of Steroidogenic Acute Regulatory (StAR) protein via cyclic AMP/protein kinase A/cyclic AMP responsive element (CRE) binding protein (CREB). CREB-dependent transcription for a number of genes requires the CREB co-activator, Transducer of Regulated CREB activity (TORC). We have recently demonstrated that nuclear translocation of TORC2 precedes transcriptional activation of StAR following ACTH injection in rats, suggesting an early involvement in transcriptional activation. To determine whether TORC is required for initiation of StAR transcription, we used the ACTH-responsive mouse adrenocortical cell line; Y1-A70, to examine the time-relationship between nuclear translocation of the TORC isoforms, TORC 1, 2 and 3 and StAR transcription (measured as changes in StAR hnRNA levels) following stimulation with ACTH, and the effect of knocking down TORC expression on ACTH-stimulated StAR transcription. Western blot analysis revealed about a 15-fold increase in nuclear dephosphorylated TORC2 by 3 min incubation with 10[sup]-8[/sup]M ACTH, reaching about 20-fold by 7 min, remaining in a plateau between 15 and 30 min and starting to decrease by 60 min to reach near basal levels by 120 min. TORC1 and TORC3 were also rapidly translocated to the nucleus at 3 min, though the fold changes were smaller than those for TORC2. In contrast, StAR hnRNA levels were unchanged by 3 min, increased progressively between 7 and 45 min, declined to about 60% of basal levels by 60 min and remained at these levels up to 120 min incubation. Knockdown of TORC2 or TORC3 or their combination using siRNA oligonucleotides, attenuated ACTH-induced increase in StAR hnRNA. The pattern of TORC translocation to the nucleus preceding the increases in StAR transcription following ACTH exposure, similar to those seen [italic]in vivo[/italic], is consistent with the possibility that all three TORC isoforms are involved in StAR transcription. The inhibitory effect of TORC2 and TORC3 knockdown on ACTH-stimulated StAR hnRNA supports the involvement of these isoforms on the transcriptional regulation of StAR. In addition, the data shows that Y1-A70 cells provide a good model for studying the stimulation of endogenous steroidogenic protein transcription by ACTH.[br][br]Sources of Research Support: Intramural Research Program, NICHD.[br][br]Nothing to Disclose: LIFS, MO, YL, GA 2012-06-24T11:30:00 310 2012-06-24T00:00:00 1899-12-30T11:30:00 1011 152 1003 OR16-2 OR03-01 Sunday 830 2012


830 ENDO12L_OR16-3 ORAL SESSION: Regulation of Adrenal Cortical Function (11:15 AM-12:45 PM) Influence of Unilateral Adrenalectomy on Set Point of Hypothalamic-Pituitary-Adrenal Axis in Long-Term Survivors of Childhood Nephroblastoma and Neuroblastoma M van Waas, R Pieters, M M van Noesel, A J van der Lely, F H de Jong, M M van den Heuvel-Eibrink, S JCMM Neggers Erasmus University Medical Centre Rotterdam, Rotterdam, Netherlands; Sophia Children[apos]s Hospital and Erasmus University Medical Centre Rotterdam, Rotterdam, Netherlands INTRODUCTION.[br]Adrenal insufficiency, or relative insufficiency, might partly explain increased mortality rates in nephroblastoma and neuroblastoma survivors after unilateral adrenalectomy. Aim of this study was to assess adrenal function and its metabolic effects after unilateral adrenalectomy.[br]PATIENTS AND METHODS[br]Cross-sectional study with a socio-demographically similar control group, conducted between October 2009-March 2011 at the Erasmus University Medical Centre, the Netherlands. Sixty-seven adult long-term survivors of nephroblastoma, 36 survivors of neuroblastoma and 49 control subjects were included in this study. Adrenal function was assessed by a 1mcg short Synacthen-test. Cortisol, adrenocorticotrophic hormone (ACTH), low (LDL-C) and high-density lipoprotein-cholesterol (HDL-C), triglycerides, apolipoprotein-B, glucose and insulin levels were assessed in blood samples taken at baseline. In addition, cortisol levels were assessed after 30 (t=30) and 60 minutes. Homeostatic Model Assessment (HOMA) was calculated.[br]RESULTS.[br]Adrenal insufficiency was not present in survivors. Interestingly, baseline serum cortisol levels were higher in survivors after unilateral adrenalectomy (mean 503 nmol/l) (N=46) than in survivors with both adrenals intact (mean 393 nmol/l, P=0.002) (N=52), and than in controls (mean 399 nmol/l, P=0.013) (N=49). After correcting for age, sex, and use of oral estrogens, unilateral adrenalectomy was independently associated with elevated baseline cortisol and ACTH levels. Baseline cortisol levels were positively associated with triglycerides (P[lt]0.001), LDL-C (P=0.004), apolipoprotein-B (P[lt]0.001) and HOMA (P=0.008).[br]CONCLUSIONS.[br]No adrenal insufficiency was observed in survivors of nephroblastoma and neuroblastoma. Survivors treated with unilateral adrenalectomy had relatively high basal cortisol and ACTH levels, indicating a higher central setpoint of the hypothalamic-pituitary-adrenal axis. This higher setpoint was associated with lipid concentrations and insulin resistance.[br][br]Sources of Research Support: KIKA.[br][br]Nothing to Disclose: MvW, RP, MMvN, AJvdL, FHdJ, MMvdH-E, SJCMMN 2012-06-24T11:45:00 310 2012-06-24T00:00:00 1899-12-30T11:45:00 57 152 1004 OR16-3 OR03-01 Sunday 831 2012


831 ENDO12L_OR16-4 ORAL SESSION: Regulation of Adrenal Cortical Function (11:15 AM-12:45 PM) Reduced Cortisol Metabolism as a Driver of ACTH Suppression during Critical Illness Eva Boonen, Hilke Vervenne, Philippe Meersseman, Leen Mortier, Yoo-Mee Vanwijngaerden, Isabel Spriet, Brian R Walker, Lies Langouche, Ilse Vanhorebeek, Greet Van den Berghe KU Leuven, Leuven, Belgium; KU Leuven, Leuven, Belgium; Jessa Hospital, Hasselt, Belgium; KU Leuven, Leuven, Belgium; University of Edinburgh, Edinburgh, UK Background: Critical illness is hallmarked by elevated circulating cortisol, which is traditionally exclusively attributed to stress-induced increased cortisol production. However, given paradoxically low ACTH, as previously reported, we hypothesized that reduced cortisol metabolism plays a key role in maintaining high cortisol during critical illness which, by negative feedback, suppresses ACTH and cortisol production.[br]Methods: In a first study (N=59), the time course of ACTH and cortisol during the first week in ICU was documented. In a second study (N=28), plasma half-life of exogenous cortisol was determined in critically ill patients. In a third study (N=51), urinary cortisol metabolites were quantified to estimate the activity of cortisol-metabolizing enzymes. In a fourth study (N=64), the major cortisol-metabolizing enzymes were quantified in liver and adipose tissue in relation to circulating cortisol and bile acids. Finally, an ongoing stable-isotope cortisol tracer study will further characterize altered cortisol metabolism and production.[br]Each study included a comparable heterogeneous ICU population and a healthy control group, matched for age, gender and BMI.[br]Results: Total circulating cortisol was consistently higher and ACTH lower in patients than controls (p[lt]0.001), confirming the ACTH-cortisol dissociation during critical illness. Cortisol half-life was 5-fold longer (p[lt]0.001) and calculated cortisol production similar in patients compared with controls. Based on urinary metabolites, cortisol production was not increased whereas activity of 5[alpha]- and 5[beta]-reductase and 11[beta]-hydroxysteroid dehydrogenase type 2 was 3- to 6-fold lower in patients than controls (p[lt]0.0001). In liver, gene and protein expression of 5[alpha]- and 5[beta]-reductase were between 4- and 20-fold reduced (p[lt]0.0001) and correlated inversely with circulating cortisol. Moreover, enzyme expression correlated inversely with circulating bile acid levels, which were elevated 14-fold in patients and have been shown to reduce expression and activity of cortisol metabolizing enzymes.[br]Conclusions: During critical illness, markedly reduced cortisol breakdown, possibly driven by bile acid-induced suppression of cortisol-degrading enzymes, maintains cortisol levels while inducing negative feedback suppression of ACTH, which may evoke adrenal atrophy. This paradigm shift may explain why patients are predisposed to secondary adrenal insufficiency in the prolonged phase of illness.[br][br]Nothing to Disclose: EB, HV, PM, LM, Y-MV, IS, BRW, LL, IV, GVdB 2012-06-24T12:00:00 310 2012-06-24T00:00:00 1899-12-30T12:00:00 949 152 1005 OR16-4 OR03-01 Sunday 832 2012


832 ENDO12L_OR16-5 ORAL SESSION: Regulation of Adrenal Cortical Function (11:15 AM-12:45 PM) Cortisol Awakening Response (CAR) and Cortisol Circadian Rhythm in Cushing Syndrome Silvia L Ruiz, Paula CL Elias, Margaret de Castro, Ayrton C Moreira School of Medicine or Ribeir[atilde]o Preto - University of S[atilde]o Paulo, Ribeir[atilde]o Preto, Brazil In healthy subjects, morning awakening is associated with a subtle burst of cortisol secretion: cortisol awakening response (CAR), a physiological response to awakening (1). CAR is a distinct component of the cortisol daily variation, with characteristics probably unrelated to those of cortisol circadian rhythm (CR) (2). The absence of CR in Cushing[apos]s Syndrome (CS) has been previously studied. However, CAR pattern in CS is still unknown. The aim of this study was to compare CAR and CR in patients with active CS, in CS patients after remission (CSR), patients with pseudo-Cushing (PS) and healthy subjects (HS).[br]We evaluated 14 HS, 5 patients with PS, 7 with CSR and 6 with CS, age and gender paired. Salivary free cortisol (SF) samples were collected (Salivette) on two consecutive days. CR was determined at 0800, 1100, 1700, 2000 and 2300h on the first day. CAR was obtained upon awakening on the next morning at 15, 30, 45 and 60 min post-awakening. SF was determined by RIA with assay sensitivity of 60 ng/dl. Kruskal-Wallis test was performed for multiple comparisons. The Wilcoxon-Mann-Whitney test and Spearman correlations were used as appropriate. Significance was assumed when p[lt] 0.05.[br]SF levels (ng/dL; mean [plusmn] SD) at 2300h during CR measurements were 216 [plusmn] 136, 190[plusmn]70, 450[plusmn]322, 1400[plusmn]540 in HS, PS, CSR and CS, respectively. Moreover, SF CAR levels increased significantly within the first 30 minutes after awakening (p[lt]0,05) in all groups, but PS group (p=0.06). At 60 minutes, SF levels had decreased progressively in all groups, but attained baseline levels only in CS group. The CAR absolute peak values were 2582[plusmn]690; 2433[plusmn]612; 2269[plusmn]743; 2798[plusmn]1166 (NS) and the relative increase in cortisol levels were 143%, 273%, 201% and 46% in HS, PS, CSR and CS, respectively (p=0.008). There was a nonsignificant correlation between CR levels at 0800h or 2300h and CAR cortisol basal or peak levels in HS, PS and CSR. However, CS group presented a positive correlation between 0800h CR level and basal and peak CAR levels (r=0.94; p=0.01).[br]Our results demonstrated the presence of CAR in HS, PS, CSR groups. CR and CAR seems to be unrelated components of the daily variation of HPA axis, in these groups. In CS patients CAR was attenuated. CAR could be blunted or masked by morning hypercortisolism in CS.[br][br]1.Wilhelm I, Psychoneuroendocrinology, 32, 358, 2007. 2.Clow A, Stress, 7, 29, 2004.[br][br]Sources of Research Support: FAPESP Grants # 2010/03039-7; 2007/58365-3 and CNPq.[br][br]Nothing to Disclose: SLR, PCLE, MdC, ACM 2012-06-24T12:15:00 310 2012-06-24T00:00:00 1899-12-30T12:15:00 334 152 1006 OR16-5 OR03-01 Sunday 833 2012


833 ENDO12L_OR16-6 ORAL SESSION: Regulation of Adrenal Cortical Function (11:15 AM-12:45 PM) Mifepristone Improves Global Clinical Status in Patients with Cushing Syndrome: Results from the SEISMIC Study Laurence Katznelson, Glenn D Braunstein, David Feldman, D Lynn Loriaux, David E Schteingart, Coleman Gross, Charissa Hogeboom Stanford University, Stanford, CA; Cedars-Sinai Medical Center, Los Angeles, CA; Oregon Health Science University, Portland, OR; University of Michigan, Ann Arbor, MI; Corcept Therapeutics, Menlo Park, CA; ICON Clinical Research, North Wales, PA [italic][underline]Rationale:[/underline][/italic] There are limited medical options for the treatment of Cushing[apos]s syndrome (CS). We present additional results on clinical response from the SEISMIC study involving use of the glucocorticoid antagonist mifepristone in CS.[br][italic][underline]Methods:[/underline] [/italic]SEISMIC was a 24 wk multicenter, open label trial of mifepristone (300-1200 mg/d) in CS patients with type 2 DM/impaired glucose tolerance (C-DM) or hypertension (C-HT). 46 patients (14M, 32F) are included in analyses of efficacy (modified intent to treat [mITT]). The key secondary endpoint was global clinical response (GCR); a 3 member data review board (DRB) considered 8 clinical data categories and graded study visits in blinded fashion as +1 (improvement), 0 (no change), -1 (worsening) vs. baseline. Positive response was defined as a median score of +1 at any postbaseline visit. A cumulative logistic threshold model was used to estimate intraclass correlation coefficient (ICC) of DRB raters.[br][italic][underline]Results:[/underline][/italic] The proportion of positive GCRs in mITT increased during the study: wk 6, 43%; wk 10, 50%; wk 16, 78%; wk 24, 83% (p[lt]0.001). GCR was 94% at any postbaseline visit in the completer group (N=33). There was 76.6% agreement between members of the DRB, with an ICC of 0.652 (moderate agreement, p[lt]0.0001). Overall, the rate of positive GCRs was similar between genders (men 79%; women 91%, p=NS). Men had a high proportion of positive GCRs by wk 6 which then plateaued (wk 6, 64%; wk 10, 57%; wk 16, 71%; wk 24, 64%); in women, there was a gradual increase in proportion of positive GCR (wk 6, 31%; wk 10, 34%; wk 16, 56%; wk 24, 75%) (repeated measures, p=0.003; 1st visit with positive GCR by gender, p=0.02); no such trend was apparent for age. By wk 24, investigator graded Cushingoid appearance was much improved in 52%, somewhat improved in 33%, and unchanged in 15% (p[lt]0.0001). Quality of life (SF-36) improved by 24 wk in the following scales (mean[plusmn]SD): General Health, 4.4[plusmn]8.3 (p=0.004); Physical Function, 7.1[plusmn]9.4 (p[lt]0.0001); Role Physical, 3.3[plusmn]10.4 (p=0.05); Social Functioning, 7.7[plusmn]11.6 (p=0.0003); Vitality, 6.3[plusmn]11.1 (p=0.002); Mental Health, 4.1[plusmn]10.5 (p=0.03); Role Emotional, 4.9[plusmn]12.4 (p=0.03).[br][underline][italic]Conclusion[/italic]:[/underline] Treatment of CS with mifepristone results in early and progressive clinical improvement in most patients, including physical appearance and enhancement of quality of life. The time to achieve GCR in men was shorter than in women, a finding that should be considered when monitoring clinical response.[br][br]Sources of Research Support: Corcept Therapeutics.[br][br]Disclosures: LK: Investigator, Novartis Pharmaceuticals, Corcept Therapeutics; Research Funding, Pfizer, Inc., Ipsen. GDB: Consultant, Corcept Therapeutics. DF: Consultant, Corcept Therapeutics. DLL: Consultant, Corcept Therapeutics. DES: Investigator, Corcept Therapeutics. CG: Employee, Corcept Therapeutics; Employee, Corcept Therapeutics. Nothing to Disclose: CH 2012-06-24T12:30:00 310 2012-06-24T00:00:00 1899-12-30T12:30:00 696 152 1007 OR16-6 OR03-01 Sunday 834 2012


834 ENDO12L_OR17-1 ORAL SESSION: Emerging Strategies in Diabetes (11:15 AM-12:45 PM) Long-Term Follow-Up of Patients with Type 1 Diabetes on Liraglutide and the Effect of Liraglutide as Additional Treatment in Obese Patients with Type 1 Diabetes Nitesh D Kuhadiya, Ritu Malik, Natalie Bellini, Jane Lyons Patterson, Andrea Traina, Antoine Makdissi, Paresh Dandona State University of New York (SUNY), Buffalo, NY; Rochester General Hospital, Rochester, NY; St John Fisher College, Rochester, NY We have recently shown that the addition of Liraglutide to insulin in the treatment of well controlled, non-obese patients with type 1 diabetes leads to a significant further rapid reduction in glycemia, glycemic excursions, HbA1c, insulin requirements and body weight within days. We now present data of 8 patients with type 1 diabetes (mean age: 43[plusmn]8 yrs; mean duration of diabetes: 29[plusmn]11 yrs) on Liraglutide treatment (mean dose: 1.72[plusmn]0.67mg) for 1.24[plusmn] 0.21 yrs. Percent time spent in hyperglycemia calculated by using 3 glycemic thresholds ([gt]150,200 and 250 mg/dl) fell by 45.35%(p = 0.01), 45.19%(p = 0.03) [amp] 61.92%(p =0.001) respectively. There was a reduction in the bolus, basal and total daily doses of insulin by 26%(p = 0.009), 19%(p = 0.01) and 20%(p = 0.008) respectively. The body weight fell from 85.3[plusmn] 12.6 to 78.7[plusmn]15.9 Kg (p = 0.01) and the BMI from 30.3 [plusmn] 3.9 to 27.8 [plusmn]4.9Kg/m2 (p = 0.01). Fasting C-peptide concentrations were non-detectable at the outset and at the end of the study. We have now also investigated the effect of the addition of 1.8 mg of Liraglutide in 15 obese patients with type 1 diabetes (10 females, 5 males, 14 Caucasian, 1 African American; mean age: 47 [plusmn]13.81 yrs; mean duration of diabetes: 20.06 [plusmn] 10.18 yrs) who were not well controlled (mean HbA1c: 7.8[plusmn]0.82). Over a period of 6 months, HbA1c fell to 7.39[plusmn] 0.77%(p = 0.05); the bolus dose of insulin decreased significantly from 35.92 [plusmn] 19.69 to 29.52 [plusmn] 17.10 units (p = 0.03) while the basal dose did not change; the body weight fell from 100.63 [plusmn] 18.31 Kg to 95.96[plusmn]19.22 kg (p = 0.008) and the BMI from 34.06[plusmn]7.36 to 32.33[plusmn]6.85 Kg/m2 (p = 0.01). The systolic BP decreased from 137.53[plusmn] 18.86 to 121.86[plusmn]13.53(p = 0.003). We conclude that Liraglutide treatment in patients with type 1 diabetes has a rapid, sustained and durable effect on glycemia, body weight, insulin dose and systolic blood pressure. In addition, poorly controlled, obese patients with type 1 diabetes also benefit from this treatment. Large, prospectively randomized studies are required to establish the use of Liraglutide in type 1 diabetes. Mechanistic studies to elucidate the mode of action of Liraglutide in these patients are also required since they do not have any beta-cell reserve.[br][br]Sources of Research Support: This research was not funded.[br][br]Disclosures: PD: Speaker, Novo Nordisk; Principal Investigator, Novo Nordisk; Advisory Group Member, Novo Nordisk. Nothing to Disclose: NDK, RM, NB, JLP, AT, AM 2012-06-24T11:15:00 Theater C 2012-06-24T00:00:00 1899-12-30T11:15:00 645 153 1008 OR17-1 OR11-01 Sunday 835 2012


835 ENDO12L_OR17-2 ORAL SESSION: Emerging Strategies in Diabetes (11:15 AM-12:45 PM) Subjects Achieving Good Glycemic Control (HbA[sub]1c[/sub] [lt]7.0%) Experience a Lower Rate of Confirmed and Nocturnal Confirmed Hypoglycemia with Insulin Degludec Than with Insulin Glargine: A Meta-Analysis of Phase 3a Trials Daniel Einhorn, Yehuda Handelsman, Bruce W Bode, Lars Endahl, Henriette Mersebach, Allen B King Scripps Whittier Diabetes Institute, La Jolla, CA; Metabolic Institute of America, Tarzana, CA; Atlanta Diabetes Associates, Atlanta, GA; Novo Nordisk A/S, Soeborg, Denmark; Novo Nordisk A/S, Soeborg, Denmark; Diabetes Care Center, Salinas, CA [bold]Background[/bold][br]Insulin degludec (IDeg) is a new basal insulin with an ultra-long, stable action profile with low hour-to-hour and day-to-day variability, which is predicted to contribute to a lower risk for hypoglycemic episodes, especially as patients approach titration targets. The objective of this meta-analysis was to compare hypoglycemia rates with IDeg vs. insulin glargine (IGlar) in subjects achieving good glycemic control (HbA[sub]1c[/sub] [lt]7% at end of trial).[br][bold]Methods[/bold][br]A pre-planned prospective meta-analysis evaluating patient data from the IDeg 3a program using a negative binomial regression model was performed in a pooled population with type 1 (T1DM) or type 2 (T2DM) diabetes to compare hypoglycemia rates with IDeg vs. IGlar. Confirmed hypoglycemia was defined as self-reported PG [lt]56 mg/dL or a severe episode requiring assistance; nocturnal confirmed hypoglycemia had onset 00:01-05:59, inclusive. The analysis included data from all open-label, randomized, treat-to-target, confirmatory 26- or 52-wk trials with IDeg (n=2899) vs. IGlar (n=1431) dosed once daily in T1DM (2 trials) and T2DM (5 trials). Data obtained during the maintenance period (defined as stable glycemia and insulin dose [obtained from 16 wks onwards]) were also analyzed. This report presents a post hoc analysis comparing confirmed and nocturnal confirmed hypoglycemia rates in subjects achieving HbA[sub]1c[/sub] [lt]7% at end of trial (IDeg, n=1347; IGlar, n=697).[br][bold]Results[/bold][br]Across trials, 47.2% of all subjects (40.0%, T1DM; 49.3%, T2DM) achieved HbA[sub]1c[/sub] [lt]7% at end of trial. Subjects with HbA[sub]1c[/sub] [lt]7% had a 14% lower rate of overall confirmed hypoglycemia (rate ratio [RR] IDeg/IGlar: 0.86 [95% CI: 0.76; 0.98], p[lt]0.05) and a 37% lower rate of nocturnal hypoglycemia with IDeg compared to IGlar (RR IDeg/IGlar: 0.63 [95% CI: 0.52; 0.77], p[lt]0.01). Data from the maintenance period showed an even greater reduction in hypoglycemia risk for IDeg vs. IGlar (21% lower rate of overall episodes (RR: 0.79 [95% CI: 0.68; 0.92], p[lt]0.05) and a 43% lower rate of nocturnal episodes (RR: 0.57 [95% CI: 0.45; 0.72], p[lt]0.01).[br][bold]Conclusion[/bold][br]Subjects achieving HbA[sub]1c[/sub] [lt]7% experienced statistically significantly lower rates of confirmed and nocturnal confirmed hypoglycemia with IDeg vs. IGlar. The pattern of reduced hypoglycemia with IDeg compared to IGlar is observed across the IDeg clinical program; this report emphasizes the consistency of this finding in patients achieving good glycemic control.[br][br]Disclosures: DE: Advisory Group Member, Novo Nordisk. YH: Consultant, Novo Nordisk; Research Funding, Novo Nordisk. BWB: Research Funding, Novo Nordisk; Consultant, Novo Nordisk; Speaker Bureau Member, Novo Nordisk. LE: Employee, Novo Nordisk. HM: Employee, Novo Nordisk. ABK: Consultant, Novo Nordisk; Research Funding, Novo Nordisk; Speaker Bureau Member, Novo Nordisk. 2012-06-24T11:30:00 Theater C 2012-06-24T00:00:00 1899-12-30T11:30:00 557 153 1009 OR17-2 OR11-01 Sunday 836 2012


836 ENDO12L_OR17-3 ORAL SESSION: Emerging Strategies in Diabetes (11:15 AM-12:45 PM) Safety and Efficacy of Linagliptin in Combination with Basal Insulin: A Pre-Specified, Pooled Analysis in a Vulnerable Population of Elderly Patients (Age [ge] 70 Years) with Type 2 Diabetes Maximilian von Eynatten, Dietmar Neubacher, Sanjay Patel, Hans-Juergen Woerle Boehringer Ingelheim, Ingelheim, Germany; Boehringer Ingelheim, Biberach, Germany; Boehringer Ingelheim, Bracknell, UK Elderly patients with type 2 diabetes (T2D) are commonly characterized by longer disease duration and diminished [beta]-cell capacity, which often requires combination therapy with basal insulin. Increasing the insulin dose is one option to improve glycemic control, but this often presents a higher risk of hypoglycemia, which increases overall safety concerns in this vulnerable population. Elderly patients may therefore benefit from an additional oral antidiabetic drug (OAD) that can be administered safely in combination with their background basal insulin. Linagliptin is a recently approved oral DPP-4 inhibitor that has shown clinically meaningful efficacy and an overall safety profile similar to placebo in commonly used OAD regimens. It also has a once-daily, single-dose strength posology without the need for dose adjustment in patients with hepatic and/or renal impairment, which allows for convenient dosing and may support patient adherence. Two phase 3 studies evaluating linagliptin vs. placebo as add-on therapy to basal insulin (NCT00954447) and as T2D management in elderly patients (NCT01084005) were eligible for a pre-specified pooled analysis exploring its safety and efficacy in combination with basal insulin in elderly patients (age [ge] 70 years) with T2D. The primary efficacy endpoint was change from baseline to week 24 in HbA1c. A total of 247 patients inadequately controlled on insulin glargine, insulin detemir, or NPH insulin received either linagliptin 5 mg once daily (n=126) or placebo (n=121). Mean insulin doses were 35.0 IU and 36.6 IU, respectively. Baseline age and HbA1c were 74.3 years and 8.2% in both groups. Overall, 95.1% of all individuals had a diabetes duration [gt] 5 years. At week 24, the placebo-adjusted mean change in HbA1c with linagliptin was [minus]0.77% (95% CI: [minus]0.95, [minus]0.59; p[lt]0.0001). Hypoglycemia occurred in 28.6% and 37.2% of linagliptin and placebo patients, respectively. The frequency of premature discontinuation was lower with linagliptin than with placebo (7.1% vs. 14.0%). The most frequent reasons were occurrence of an adverse event (linagliptin: 4.0% vs. placebo: 3.3%) and refusal to continue trial medication (linagliptin: 1.6% vs. placebo: 5.0%). In conclusion, in this vulnerable elderly population, linagliptin in combination with basal insulin was well tolerated and achieved clinically meaningful improvements in glycemic control without excessive risk of hypoglycemia.[br][br]Sources of Research Support: Boehringer Ingelheim.[br][br]Disclosures: MvE: Employee, Boehringer Ingelheim. DN: Employee, Boehringer Ingelheim. SP: Employee, Boehringer Ingelheim. H-JW: Employee, Boehringer Ingelheim. 2012-06-24T11:45:00 Theater C 2012-06-24T00:00:00 1899-12-30T11:45:00 804 153 1010 OR17-3 OR11-01 Sunday 837 2012


837 ENDO12L_OR17-4 ORAL SESSION: Emerging Strategies in Diabetes (11:15 AM-12:45 PM) Increased Risk of Overall Mortality in Patients with Type 2 Diabetes Receiving Glipizide, Glyburide, and Glimepiride vs. Metformin. A Retrospective Analysis Kevin M Pantalone, Michael W Kattan, Changhong Yu, Brian J Wells, Susana Arrigain, Anil Jain, Ashish Atreja, Robert S Zimmerman Summa Western Reserve Hospital Physicians, Inc, Hudson, OH; Cleveland Clinic, Cleveland, OH; Cleveland Clinic, Cleveland, OH; Cleveland Clinic, Cleveland, OH; Cleveland Clinic, Cleveland, OH [bold]Background[/bold]: It remains uncertain if differences in mortality risk exist among the sulfonylureas, especially in patients with documented coronary artery disease (CAD). Conflicting reports have surfaced concerning whether an increased overall mortality (or cardiovascular mortality) risk accompanies the various sulfonylureas (1-6). A difference in mortality risk may exist, as the individual sulfonylureas vary significantly from one another in terms of hypoglycemic risk, sulfonylurea receptor selectivity, and their effects on myocardial ischemic preconditioning (7, 8).[br][bold]Purpose:[/bold] To assess the risk of overall mortality in patients with type 2 diabetes treated with glipizide, glyburide, or glimepiride vs. metformin.[br][bold]Methods:[/bold] A retrospective cohort study was conducted using an academic health center enterprise-wide electronic health record (EHR) system to identify 23,915 patients with type 2 diabetes who initiated monotherapy with metformin (N=12,774), glipizide (N=4,325), glyburide (N=4,279), or glimepiride (N=2,537), [ge] 18 years of age, with and without a history of CAD. The patients were followed for mortality by documentation in the EHR and Social Security Death Index. Multivariable Cox models with propensity analysis were used to compare the anti-diabetic monotherapies.[br][bold]Results:[/bold] Baseline characteristics for the entire cohort and subgroup of patients with CAD include mean age (years) +/- SD of 61.8 +/- 14.3 and 67.9 +/- 11.0 years, 50.4% and 69% male, and 77.9% and 84% Caucasian, respectively. The median follow-up was 2.2 years. There were a total of 2,546 deaths in 58,513 person years of follow-up in the entire cohort, and 419 deaths in 5,980 person years of follow-up in the subgroup of patients with documented CAD. An increase in overall mortality risk was observed in the entire cohort with glipizide (HR 1.64; 95% CI 1.39-1.94), glyburide (HR 1.59; 95% CI 1.35-1.88), and glimepiride (HR 1.68; 95% CI 1.37-2.06) vs. metformin; however, in those patients with documented CAD, a statistically significant increase in overall mortality risk was only found with glipizide (HR 1.41; 95% CI 1.07-1.87) and glyburide (HR 1.38; 95% CI 1.04-1.83) vs. metformin.[br][bold]Conclusions: [/bold]Glipizide, glyburide, and glimepiride are associated with an increased risk of overall mortality vs. metformin. Our results suggest that if a sulfonylurea is required to obtain glycemic control, glimepiride may be the preferred sulfonylurea in those with underlying CAD.[br][br]1. Khalangot M et al.,Diabetes Res Clin Pract 2009 Dec;86(3):247-53. 2. Pantalone KM et al.,Diabetes Care 2010 Jun;33(6):1224-1229. 3. Schramm TK et al., Eur Heart J. 2011 Aug;32(15):1900-8. 4. Zeller M et al., J Clin Endocrinol Metab. 2010 Nov;95(11):4993-5002. 5. Horsdal HT et al., Diabetes Metab Res Rev 2009 Sep;25(6):515-522. 6. Andersson C et al., Diabetes Res Clin Pract 2011 Oct;94(1):119-125. 7. Gangji AS et al., Diabetes Care 2007 Feb;30(2):389-394. 8. Krentz AJ., International Congress Series 2003;1253:261-277.[br][br]Sources of Research Support: This research was supported through a grant from Astra Zeneca.[br][br]Disclosures: RSZ: Speaker Bureau Member, Novo Nordisk, Santarus, Inc.; Consultant, Bristol-Myers Squibb, Astra Zeneca; Research Funding, Merck & Co. Nothing to Disclose: KMP, MWK, CY, BJW, SA, AJ, AA 2012-06-24T12:00:00 Theater C 2012-06-24T00:00:00 1899-12-30T12:00:00 691 153 1011 OR17-4 OR11-01 Sunday 838 2012


838 ENDO12L_OR17-5 ORAL SESSION: Emerging Strategies in Diabetes (11:15 AM-12:45 PM) Addition of Exenatide or Sitagliptin to Insulin in Patients of New-Onset Type 1 Diabetes KVS Hari Kumar, P Prusty Command Hospital Central Command, Lucknow, India Objective [ndash] Incretin based therapies are known to have pleotropic benefits in diabetes mellitus including preservation of [beta]-cell mass in animal studies suggesting a definite role in type 1 diabetes (1). To study the effect of addition of incretin modulators to the insulin requirement in new onset type 1 diabetes mellitus (T1DM).[br]Methods [ndash] Fifteen newly detected T1DM adult patients participated in this open label, randomized study for a period of one year. The patients were divided into 3 groups: Group1 (Insulin alone), Group 2 (Insulin and Exenatide), Group 3 (Insulin and Sitagliptin). The primary outcome was change in insulin requirement from onset of T1DM and secondary outcomes were preservation of c peptide secretion and risk of hypoglycemia at the end of one year observation (NCT01235819).[br]Results [ndash] The study participants consist of 12 males and 3 females with a mean age of 26.8 [plusmn] 3.5 yr. The decrease in insulin requirement was 16.7 [plusmn] 12.5, 39.8 [plusmn] 17.2, 21.2 [plusmn] 9.6 units in groups 1, 2 and 3 respectively at the end of one year (P=0.0431). The mean stimulated c peptide secretion was 0.34 [plusmn] 0.12, 0.45 [plusmn] 0.34, 0.44 [plusmn] 0.5 ng/mL at the end of the study period in 3 groups respectively (P=0.8656). Group 2 had the maximum percentage preservation in c peptide when compared with other groups. Two patients in group 2 and one patient in group 3 did not require insulin during the last 9 months. The incidence of severe hypoglycemia was same in all the groups and none of the study participants had ketoacidosis during follow up.[br]Conclusions [ndash] Our preliminary data showed that the addition of exenatide at onset in patients of T1DM decreases the insulin requirement. Use of incretin modulators had no effect on endogenous insulin production and risk of hypoglycemia.[br][br](1) Bosi E. J Clin Endocrinol Metab 2010;95:2607-9.[br][br]Nothing to Disclose: KVSHK, PP 2012-06-24T12:15:00 Theater C 2012-06-24T00:00:00 1899-12-30T12:15:00 573 153 1012 OR17-5 OR11-01 Sunday 839 2012


839 ENDO12L_OR17-6 ORAL SESSION: Emerging Strategies in Diabetes (11:15 AM-12:45 PM) Metformin and Sulfonylureas in Relation to Cancer Risk in Diabetic Patients: A Systematic Review and Meta-Analysis Bindiya Thakkar, Konstantinos N Aronis, Maria T Vamvini, Christos S Mantzoros Boston VA Healthcare System, Harvard Medical School, Boston, MA; Boston Medical Center, Boston University School of Medicine, Boston, MA; Beth Israel Deaconess Medical Center, Boston, MA Aim: Accumulating evidence suggests that patients with type 2 diabetes mellitus and hyperinsulinemia are at increased risk for developing cancers. It remains to be fully elucidated whether use of Metformin, an insulin sensitizer, and/or Sulfonylureas, insulin secretagogues, affect cancer incidence.[br]Methods: We performed a systematic review and meta-analysis on this topic. A comprehensive literature search using PubMed was performed to identify randomized control trials (RCTs), cohorts and case-control studies published until January 2012 that assess the effects of metformin and/or sulfonylureas on cancer incidence in diabetic patients. Seventeen studies on Metformin and fifteen studies on Sulfonylurea were included in the meta-analysis. Fixed and Random effects meta-analysis models were utilized, following the Mantel-Haenszel and the DerSimonian Laird methods accordingly. The effect size was summarized as risk ratio (RR) for RCTs and cohorts and as odds ratio (OR) for the cross-sectional studies. Heterogeneity of the outcomes was evaluated with the I-squared statistic (I2). Analysis for the existence of potential publication bias was performed with Egger[apos]s linear regression for funnel plot asymmetry and correction of this bias was performed with the trim-and-fill method of Duval [amp] Tweedie. Statistical analysis was performed in R v2.13.[br]Results: On the basis of data from a total of 92,570 subjects followed for 54.92[plusmn]19.70 weeks in both cohort studies and RCTs and 31,136 subjects in case control studies, we found that use of Metformin is associated with a clinically and statistically significant reduction in cancer risk in both cohort studies and RCTs (RR=0.62 [95%CI= 0.59-0.66], I2=97.8%, P[lt]0.001) and case controls studies (OR=0.78 [95%CI= 0.72-0.85], I2=83.9%, P[lt]0.001). Data from a total of 135,615 subjects followed for 50.82[plusmn]14.24 weeks in both cohort studies and RCTs shows that use of Sulfonylureas is associated with an increase in the cancer risk in these studies (RR=1.48 [95%CI= 1.40-1.56], I2=95.6%, P[lt]0.001 whereas data from case control studies with 17,475 subjects failed to demonstrate the same effect (OR=1.06 [95%CI= 0.92-1.22], I2=62.3%, P = 0.39).[br]Conclusions: Use of Metformin reduces, while use of Sulfonylureas appears to increase the risk for cancer in diabetics. These data need to be confirmed in large scale RCTs before translating these results into clinical practice.[br][br]Nothing to Disclose: BT, KNA, MTV, CSM 2012-06-24T12:30:00 Theater C 2012-06-24T00:00:00 1899-12-30T12:30:00 1031 153 1013 OR17-6 OR11-01 Sunday 840 2012


840 ENDO12L_OR18-1 ORAL SESSION: Adiposity [amp] Its Sequelae (11:15 AM-12:45 PM) Adipose Tissue Dysfunction in Humans with the Metabolic Syndrome Demidmaa Tuvdendorj, Elizabeth Murphy, Manisha Chandalia, Nicola Abate Univesity of Texas Medical Branch, Galveston, TX; University of San Francisco, San Francisco, CA [bold]Introduction:[/bold] Obesity is associated with metabolic abnormalities, i.e. insulin resistance/metabolic syndrome (IR/MS). However, not all obese people develop metabolic complications. The mechanisms of different metabolic effect of weight excess are focus of intense investigation. In this study we evaluated the role of differences in adipose tissue function between normoglycemic volunteers with and without the metabolic syndrome. We measured synthesis rate of triglyceride (TG) in abdominal subcutaneous adipose tissue using deuterium labeling approach and the gene expression of lipoprotein lipase (LPL) and hormone sensitive lipase (HSL) in two groups of subjects [ndash] with metabolic syndrome (MS) and without MS (NMS). [bold]Research Design and Methods:[/bold] The subjects (NMS (n=7) [italic]vs[/italic]. MS (n = 6); Age: 31[plusmn]8 [italic]vs. [/italic]52[plusmn]26 years, [italic]p[/italic] [gt] 0.05; BMI: 29[plusmn]6 [italic]vs.[/italic] 34[plusmn]3, p [gt] 0.05, Mean[plusmn]SD) ingested deuterium water for 12 weeks; they then underwent abdominal subcutaneous adipose tissue biopsy. Adipose tissue triglyceride was extracted, and incorporation of deuterium label into the glycerol was measure using gas-chromatography-mass-spectrometer. The fractional synthesis rate (FSR) of adipose tissue TG was calculated using MIDA approach. Gene expressions of LPL and HSL were evaluated using RT-PCR. [bold]Results:[/bold] The TG-FSR normalized for body fat content was significantly lower in the MS group compared to the NMS ([italic]p [/italic]= 0.007). LPL and HSL gene expression was also significantly suppressed in the MS group compared to the NMS group (LPL, [italic]p[/italic] = 0.001, HSL, [italic]p[/italic] = 0.001). Body fat content was unchanged during the study period. Conclusion: Our results show that in humans with MS, both the synthesis and breakdown of adipose tissue TG are suppressed, suggesting that adipose tissue of obese normoglycemic subjects with MS is more [ldquo]quiescent[rdquo] than in normoglycemic subjects without the MS, independently of obesity. Further studies will confirm the role of adipose tissue metabolic quiescence as a cause of ectopic fat deposition, systemic insulin resistance and clustering of metabolic abnormalities observed in the MS.[br][br]Nothing to Disclose: DT, EM, MC, NA 2012-06-24T11:15:00 332 2012-06-24T00:00:00 1899-12-30T11:15:00 2349 154 1014 OR18-1 OR28-02 Sunday 841 2012


841 ENDO12L_OR18-2 ORAL SESSION: Adiposity [amp] Its Sequelae (11:15 AM-12:45 PM) Adipose Cell Size and Distribution: Relationship to Insulin Resistance Tracey McLaughlin, Cindy Lamendola, Teresa C Liu, Arthur Sherman, Samuel W Cushman Stanford University, Stanford, CA; National Institutes of Health, Bethesda, MD Enlargement of adipose cells has been linked to increased BMI and hyperinsulinemia. It has been proposed that impaired recruitment of new adipose cells as body fat mass expands leads to hypertrophy of existing cells and insulin resistance(IR) via FFA redistribution. The degree to which fat expansion beyond normal body weights requires adipose cell recruitment vs enlargement is not clear. Also unclear is whether adipose cell size is independently associated with insulin resistance since older studies relied on [ldquo]mean[rdquo] adipose cell size and newer techniques reveal both large/small adipose cells within a given tissue sample. We tested the hypothesis that impaired adipogenesis is associated with insulin resistance by quantifying adipose cell size distribution in 148 healthy overweight subjects in whom insulin-mediated glucose uptake was measured.[br][bold]Methods[/bold]: Periumbilical subcutaneous adipose tissue biopsy was performed on healthy volunteers with BMI 25-38 kg/m2. Adipose cell size distribution was obtained via Beckman Coulter Multisizer: primary endpoints were mean diameter of large cells and relative number of small cells. Insulin sensitivity was quantified by steady-state plasma glucose(SSPG), in which higher steady-state glucose values during standardized insulin infusion indicate resistance to insulin-mediated glucose uptake. Cell size and metabolic parameters were compared by multiple linear regression and subgroup comparison for IR/IS and body fat quintile.[br][bold]Results[/bold]: Mean age, BMI, and sex were 52[plusmn]9yrs, 31[plusmn]3 kg/m2, 61% female. Both large and small adipose cells were present in nearly equal proportions. Diameter of large adipose cells was independently associated with %body fat (r=0.26, p=0.014), female sex (r=0.21,p=0.036), and SSPG (r=0.20,p=0.012). Percent small cells was associated only with SSPG (r=0.26, p=0.003). In the highest vs lowest body fat quintile, despite a 50% difference in body fat, cell size increased by only 7% whereas cell number increased by 74%. IR individuals demonstrated a decreased proportion and number of large adipose cells but greater large cell diameter compared with body-fat matched IS individuals.[br][bold]Conclusion[/bold]: Recruitment of adipose cells is required for expansion of body fat mass beyond BMI of 25kg/m2. Insulin resistance is associated with accumulation of small adipose cells and enlargement of large adipose cells. These data support the notion that impaired adipogenesis may underlie insulin resistance in association with obesity.[br][br]1. McLaughlin T, Sherman A, Tsao P, et al. Enhanced proportion of small adipose cells in insulin-resistant vs insulin-sensitive obese individuals implicates impaired adipogenesis. Diabetologia 2007, 50:1707-15. 2. Pei, D, Jones, CNO, Bhargava, R., Chen, Y-DI, Reaven, GM. Evaluation of octreotide to assess insulin-mediated glucose disposal by the insulin suppression test. Diabetologia 1994, 37:843-5.[br][br]Sources of Research Support: National Institutes of Health/National Institute of Digestive Diseases and Diabetes, R01 DK080436, R01DK071309.[br][br]Nothing to Disclose: TM, CL, TCL, AS, SWC 2012-06-24T11:30:00 332 2012-06-24T00:00:00 1899-12-30T11:30:00 2268 154 1015 OR18-2 OR28-02 Sunday 842 2012


842 ENDO12L_OR18-3 ORAL SESSION: Adiposity [amp] Its Sequelae (11:15 AM-12:45 PM) Experimental Hyperleptinemia Acutely Increases Vasoconstrictory Sympathetic Nerve Activity in Humans Felix Machleidt, Paul Simon, Alexander Krapalis, Alexander Iwen, Manfred Hallschmid, Hendrik Lehnert, Friedhelm Sayk University Hospital Luebeck, Luebeck, Germany; University of Luebeck, Luebeck, Germany Background: Obesity and arterial hypertension are tightly connected. Hypertensive obese individuals show a significant elevation of vasoconstrictory sympathetic nerve activity (SNA). Leptin seems to play a key role in mediating these effects: 1.It is synthesized in direct proportion to body fat mass and 2. activates central sympathetic outflow via hypothalamic melanocortin receptors (MC4R) in animal models. Evidence for a causal link between hyperleptinemia, sympathetic activation and obesity-related hypertension in humans has not yet been established. The aim of this study was to characterize the effects of acute hyperleptinemia on SNA in healthy male humans. [br]Methods: In a randomized, double blind cross-over design 12 healthy male subjects with a BMI range from 20-25 kg/m2 received an intravenous bolus of either 100 [micro]g/kg body weight of recombinant human Leptin (Globe Laboratories, Canada) or placebo. Muscle sympathetic nerve activity (MSNA, [bursts per minute]) as a measure of efferent vasoconstrictory SNA was continuously recorded. 10 minute periods were analysed before (t1) and at time-points 20 min (t2), 60 min (t3) and 140 min (t4) after administration of leptin or placebo. Blood pressure and heart rate were recorded simultaneously.[br]Results: Baseline values of MSNA, blood pressure and heart rate were comparable in both groups (MSNA: t1: 24.3 [plusmn] 1.6 vs. 22.7 [plusmn] 1.7, n.s.). After application of intravenous leptin, MSNA showed a rapid and significant activation (t2: 31.0 [plusmn] 1.9 vs. 24.9 [plusmn] 1.8, p=0.05). This effect was sustained until the end of the experiment (t3: p=0.002; t4: p=0.004). There were no significant changes in blood pressure and heart rate in both groups.[br]Conclusions: Acute experimental hyperleptinemia has significant central excitatory effects on vasoconstrictory sympathetic outflow as measured by MSNA in healthy humans. These results clearly suggest that leptin functions as an important mediator linking obesity to elevated SNA and potentially to obesity related hypertension.[br][br]Nothing to Disclose: FM, PS, AK, AI, MH, HL, FS 2012-06-24T11:45:00 332 2012-06-24T00:00:00 1899-12-30T11:45:00 1837 154 1016 OR18-3 OR28-02 Sunday 843 2012


843 ENDO12L_OR18-4 ORAL SESSION: Adiposity [amp] Its Sequelae (11:15 AM-12:45 PM) Effect of a 12-Month Lifestyle Behavioral Modification Intervention on Fat Compartmentation in Overweight and Obese Prepubertal Children Rachana Dahiya, Gary J Cowin, Zhengyi Yang, May Almestehi, Gary M Leong Mater Children[apos]s Hospital, South Brisbane, Australia; The University of Queensland, St Lucia, Australia; The University of Queensland, St Lucia, Australia; The University of Queensland, St Lucia, Australia [bold]Background[br][/bold]Accumulation of fat in sites other than subcutaneous adipose tissue (SAT) has been observed in both adults and children who are obese and is associated with obesity-related comorbidities. Visceral adipose tissue (VAT) and intrahepatic lipids (IHL) are linked with increased cardiovascular disease and type 2 diabetes in adults. We therefore sought to determine whether a 12 month lifestyle behavioral modification program would decrease ectopic fat in a cohort of overweight and obese prepubertal children.[br][bold]Methods[br][/bold]20 overweight and obese children (8.7[plusmn]1.4 yrs, Male:Female, 10:10; BMI [gt]85th percentile for age and gender) and 17 lean age and gender-matched controls (8.9[plusmn]1.3 yrs, M:F, 9:8) underwent magnetic resonance imaging (MRI) and proton spectroscopy ([sup]1[/sup]H-NMR) from a group that was recruited for the KOALA Child Obesity program (1). Obese and overweight children were randomized to either Arm A: Standard care (M:F, 3:7) or Arm B: Intervention arm (M:F, 7:3) which consisted of a Scouts family camp program, extensive dietician input and Lifestyle Triple P positive parenting program over 20 weeks (2). At baseline and 12 months, height, weight, and waist circumference (WC) were measured along with whole body MRI and [sup]1[/sup]H-NMR. A semi-automated method was applied to quantify VAT and SAT at L4-5 in each participant. All spectra for IHL were fitted and analysed by using jMRUI-version 3 software.[br][bold]Results[br][/bold]Overweight and obese children had significantly higher VAT (49[plusmn]31 vs 12[plusmn]5 cm[sup]2[/sup]) and SAT (279[plusmn]68 vs 73[plusmn]45 cm[sup]2[/sup]) than the non-overweight children (p [lt]0.001). Levels of IHL were also significantly higher in overweight and obese subjects compared with lean controls (4.38[plusmn]4.1 vs 1.12[plusmn]0.7 % water peak; p[lt]0.05). A significant drop in BMI SDS (2.14[plusmn]0.4 vs 1.88[plusmn]0.4), WC SDS (2.89[plusmn]0.5 vs 2.63[plusmn]0.6)and VAT (48[plusmn]21 vs 41[plusmn]20 cm[sup]2[/sup]) was observed in the intervention group, Arm B (all p[lt]0.05), whereas subjects in Arm A had increases in SAT (279[plusmn]61 vs 352[plusmn]95 cm[sup]2[/sup]; p[lt]0.05), WC SDS (2.65[plusmn]0.5 vs 2.92[plusmn]0.3; p[lt]0.05) but no change in BMI SDS or VAT. In male subjects in Arm A, IHL significantly increased (4.1[plusmn]2.7 vs 5.4[plusmn]2.7 % water peak; p[lt]0.001) but no difference was seen in Arm B.[br][bold]Conclusion[br][/bold]Treatment of childhood obesity with an integrated family-based multi-disciplinary approach such as the KOALA Child Obesity program has demonstrated reductions in BMI SDS and VAT and prevented an increase in IHL over 12 months and thus may prevent the onset of obesity-related comorbidities.[br][br](1)Smibert, A. et al.,Eur Phys Ed Review 2010, 16 (2): 155-170. (2)West, F. et al., Behav Res Ther. 2010 Dec;48(12):1170-9.[br][br]Sources of Research Support: Golden Casket Medical Foundation; The Mazda Foundation; Merrill. Lynch Award; Golden Casket Mater Children[apos]s Hospital Funds.[br][br]Nothing to Disclose: RD, GJC, ZY, MA, GML 2012-06-24T12:00:00 332 2012-06-24T00:00:00 1899-12-30T12:00:00 1812 154 1017 OR18-4 OR28-02 Sunday 844 2012


844 ENDO12L_OR18-5 ORAL SESSION: Adiposity [amp] Its Sequelae (11:15 AM-12:45 PM) Increasing Adiposity Adversely Affects Bone Mineral Density in Elderly Obese Men Lina E Aguirre, Vibhati Kulkarny, Sheryl Pascual, Clifford R Qualls, Dennis T Villareal, Reina C Armamento-Villareal New Mexico VA Healthcare System, Albuquerque, NM; Biomedical Institute of New Mexico, Albuquerque, NM; Veterans Health Administration Affairs, Albuquerque, NM; University of New Mexico School of Medicine, Albuquerque, NM [bold]Background[/bold]: Obesity and osteoporosis are major health problems worldwide and are associated with multiple comorbidities. Traditionally, males tend to have higher bone mineral density (BMD) and lower fat mass than females and are thought to be more protected against osteoporosis. Also, it has been proposed that obesity is protective against osteoporotic fractures, yet data from epidemiological have given conflicting results.[br][bold]Purpose:[/bold] To investigate the associations between fat and bone mineral density in frail, elderly adults.[br][bold]Methods: [/bold]Baseline data from 133 frail elderly adults (58 males and 75 females) participating in a lifestyle intervention (weight loss and exercise) were analyzed. Body composition and bone mineral density were obtained using Dual-energy X-ray absorptiometry (DXA). Adipokines and inflammatory markers were measured using ELISA. HOMA-IR scores were calculated for all participants from fasting glucose and insulin levels. For all analysis, subjects were divided according to gender and according to tertiles of percent body fat (1[sup]st[/sup] tertile=lowest % fat, 3rd tertile=highest % fat).[br][bold]Results:[/bold] There were no significant differences in the baseline characteristics between genders except that females had higher average percent fat mass than males (44.80% vs. 39.82%, p=0.000, respectively). Percent fat mass was significantly negatively correlated with BMD at all body sites in men but not in women. Men in the highest tertile of % fat mass had significantly lower BMD in all the skeletal sites compared to those in the lowest tertile. This relationship was not found in women. BMD in men also correlated with adiponectin, leptin, IL-6, CRP, HOMA-IR, and osteocalcin. Multiple regression analysis showed HOMA to be a significant independent predictor of BMD in all the femoral skeletal sites and whole body BMD.[br][bold]Conclusions: [/bold]In frail, elderly adult men, increasing fat mass was negatively correlated with bone mineral density. Our data suggest that in this population obesity was not protective against osteoporosis and that males, in particular, were more vulnerable to the adverse metabolic impact of obesity on skeletal health.[br][br]Nothing to Disclose: LEA, VK, SP, CRQ, DTV, RCA-V 2012-06-24T12:15:00 332 2012-06-24T00:00:00 1899-12-30T12:15:00 1808 154 1018 OR18-5 OR28-02 Sunday 845 2012


845 ENDO12L_OR18-6 ORAL SESSION: Adiposity [amp] Its Sequelae (11:15 AM-12:45 PM) Genome-Wide Association Study of Sarcopenic-Obesity, Identifies Novel Candidate Genes. The Framingham Study Yi-Hsiang Hsu, Elizabeth Newton, Robert McLean, Douglas Kiel Hebrew SeniorLife, Boston, MA; Harvard Medical School, Boston, MA; Harvard School of Public Health, Boston, MA Sarcopenic-obesity (SO) is a condition characterized by both excess body fat and decreased muscle mass and strength. it is a common condition affecting [gt] 12% of U.S. adults aged 60 years and older. With the prolonging lifespan and an epidemic of overweight, the SO has become a major public health burden. Although obesity is a known risk factor for several adverse health outcomes, there is emerging evidence that SO has an even greater risk of cardiovascular diseases, metabolic disorders, mobility impairment and disability compared to older adults with obesity alone. Identifying genetic risk factors for SO may help to understand the pathophysiology and identify new molecular targets for treatments. We have undertaken a GWAS in 3763 Framingham participants (1578 men and 2185 women aged [gt] 50 years) using [sim] 2.6 million SNPs under an additive model adjusted for age, height, study cohorts and physical activities. To identify unique genetic determinants that associated with SO only, but not associated with obesity or sarcopenia alone, we applied a mixed-effect multinomial-logit model, which takes into account the relation of obesity, sarcopenia and SO. The SO was defined by two most common definitions: (1) muscle mass (appendicular lean mass normalized for height) and (2) muscle strength (grip strength). By muscle mass-based definition, we classified 11%, 48% and 10% of participants with SO, overweight alone and sarcopenia alone, respectively. By muscle strength-based definition, we classified 10%, 47% and 7% of participants with SO, overweight alone and sarcopenia alone, respectively. We found several SNPs associated with SO only and achieved genome-wide significance, i.e. SNPs on Chr4q22-q25 located in MANBA gene; on Chr6p12.3 near PGK2; and on Chr2q34 near FN1 gene. Several loci achieved genomic-wide suggestive level, such as SNPs in PTPRD, CDK14 and IMMP2L genes. Of note, few SNPs associated with SO were also reported to be associated with diabetes, restless leg syndrome or atherosclerosis. SNPs significantly associated with obesity were not found to be associated with SO in our analyses. The top significant SNPs (association tests p-value [lt] 5x10-6) are being further evaluated in independent studies. In conclusion, our results reveal novel candidate genes that only associated with sarcopenic-obesity, but not obesity or sarcopenia alone, which suggest potential genetic regulation of the interaction between muscle and fat energy metabolism.[br][br]Nothing to Disclose: Y-HH, EN, RM, DK 2012-06-24T12:30:00 332 2012-06-24T00:00:00 1899-12-30T12:30:00 2357 154 1019 OR18-6 OR28-02 Sunday 846 2012


846 ENDO12L_OR19-1 ORAL SESSION: Late Breaker Oral Session (11:15 AM-12:45 PM) New Metastatic Murine Prostate Cancer Cell Lines with the Genetic Characteristics of Human Cancer Xiaoming Ju, Adam Ertel, Zuoren Yu, Paolo M Fortina, Richard G Pestell Thomas Jefferson University, Philadelphia, PA; Thomas Jefferson University, Philadelphia, PA; Kimmel Cancer Center, Philadelphia, PA Defining oncogene-specific molecular pathways required for the induction and maintenance of prostate cancer is essential for the rational design of new therapies. Identifying molecular predictors of prostate cancer patient survival is important for patient therapeutic substratification. We generated murine prostate cancer cell lines via selective transduction with a single oncogene (c-Myc, Ha-Ras, NeuT, and v-Src). These cell lines demonstrate contact-independent growth characteristics in soft agar, and have the substantial growth advantage over primary prostate epithelial cells. Each of the prostate lines grew tumor in immune competent mice after subcutaneous injection, and develop lung metastasis in c-Myc, Ha-Ras, and v-Src cell line groups. RNA samples were prepared from the oncogene transformed PEC cell lines, Microarray analysis identified a total of 2635 out of 22115 genes that were significantly altered in expression (at least two-fold change) in oncogene over-expressing cell lines when compared with prostate epithelial cell control samples. The distinct oncogene expression signature was compared to gene signatures obtained from other published databases to identify similarities to human prostate cancer phenotypes. Comparisons were performed against gene signatures representative of differential expression in advanced state vs. early stage prostate cancer, high grade vs low grade prostate cancer, recurrent vs. nonrecurrent, the results indicate a significant degree of similarity between the prostate oncogene expression signature and high-grade disease and between the oncogene expression signature and the advanced stage disease phenotype. Prostate cancer Interrogation of two distinct cohorts of (297) patient samples using the oncogene signature demonstrated an ability to distinguish tumor from normal prostate with a predictive value for prostate cancer of 98 [ndash] 99%. The signature provided independent substratification of reduced recurrence free survival by Kaplan-Meier analysis. The generation of new oncogene-specific prostate cancer cell lines that recapitulate human prostate cancer gene expression, that metastasize in immune-competent mice, are a valuable new resource for testing targeted therapy. The molecular signatures identified herein provide further value over current markers of prediction and outcome.[br][br]Sources of Research Support: NIH Grant R01CA70896, R01CA75503, R01CA107382, R01CA86072 awarded to R.G.P.[br][br]Nothing to Disclose: XJ, AE, ZY, PMF, RGP 2012-06-24T11:15:00 351 2012-06-24T00:00:00 1899-12-30T11:15:00 3030 155 1020 OR19-1 OR_LB Sunday 847 2012


847 ENDO12L_OR19-2 ORAL SESSION: Late Breaker Oral Session (11:15 AM-12:45 PM) Modulation of Fibroblast Growth Factor 21 (FGF21) Diurnal Rhythm in Humans by Mild Cold Exposure: Relationship between FGF21 Levels, Lipolysis and Cold-Induced Thermogenesis Paul Lee, Robert J Brychta, Sheila Smith, Joyce Linderman, Kong Chen, Francesco Celi National Institutes of Health, Bethesda, MD Fibroblast growth factor 21 (FGF21) is a newly-discovered hormone with broad metabolic actions protecting rodents from obesity. It exhibits a diurnal rhythm in response to environmental and nutrient cues. Cold stimulates FGF21 secretion from brown adipose tissue leading to browning of white fat and enhanced adaptive thermogenesis in animals (1). In humans, the effects of cold exposure on circulating FGF21 levels, its diurnal rhythm, and its thermogenic actions have not been investigated.[br]We have evaluated the effects of mild cold exposure on FGF21 diurnal rhythm and its relationship with energy metabolism, substrate utilization and hormonal axes in healthy volunteers. Six volunteers (25[plusmn]1 yrs old, 24[plusmn]1 kg/m[sup]2[/sup], F=2) participating in a randomized, single-blind, cross-over study (2), were exposed to either ambient (24[deg]C) or mild cold (19[deg]C) temperature for 12 hours in a whole room indirect calorimeter and underwent continuous energy expenditure (EE) measurement, hormonal profiling and adipose tissue microdialysis for glycerol quantification.[br]At 24[deg]C, plasma FGF21 exhibited a diurnal rhythm, peaking at 0800 (115[plusmn]13 pg/ml); progressively dropped to a nadir at 1700 (58[plusmn]16 pg/ml, p[lt]0.05) before rising at 1900 (94[plusmn]21 pg/ml, p[lt]0.05). No changes in FGF21 levels were observed after a standardized meal at 1400.[br]Cold exposure at 19[deg]C augmented FGF21 levels and lessened the observed diurnal reduction of FGF21 at 24[deg]C from 0800-1700 by 37-68% (p[lt]0.05). Cold-induced increase in EE tended to be greater in subjects with higher plasma FGF21 concentrations (R2=0.41, p=0.08). Area under the curve (AUC) of plasma FGF21 correlated positively with AUCs of plasma non-esterified free fatty acid (R2=0.68, p[lt]0.05) and adipose microdialysate glycerol (R2=0.76, p[lt]0.05), independent of age, gender and fat mass. FGF21 levels were positively associated with urinary norepinephrine (R2=0.76, p[lt]0.05) but negatively associated with free cortisol excretion (R2=0.55, p[lt]0.05).[br]In summary, mild cold exposure augmented circulating FGF21 levels and modulated its diurnal rhythm. Upon cold exposure, FGF21 levels predicted greater fat utilization, cold-induced thermogenesis and sympathetic drive, while lessening glucocorticoid activation. From this human study, we observed that a small reduction in environmental temperature was able to modulate FGF21 diurnal rhythm with possible beneficial effects on whole body energy metabolism, mimicking pharmacological benefits of FGF21 in obese rodent studies.[br][br](1) Fisher FM et al. FGF21 regulates PGC-1[alpha] and browning of white adipose tissues in adaptive thermogenesis. Genes Dev. 2012;26:271-81[br](2) Celi FS et al. Minimal changes in environmental temperature result in a significant increase in energy expenditure and changes in the hormonal homeostasis in healthy adults. Eur J Endocrinol. 2010; 163 :863-72.[br][br]Nothing to Disclose: PL, RJB, SS, JL, KC, FC 2012-06-24T11:30:00 351 2012-06-24T00:00:00 1899-12-30T11:30:00 3040 155 1021 OR19-2 OR_LB Sunday 848 2012


848 ENDO12L_OR19-3 ORAL SESSION: Late Breaker Oral Session (11:15 AM-12:45 PM) Clinical Characteristics Predicting Beta Cell Recovery after Diabetic Ketoacidosis: Ketosis Prone Type 2 Diabetes [ndash] A Ten Year Retrospective Review Jane Weinreb, Anna Milanesi, Gelsey Goodstein VA Greater Los Angeles Medical Center, Los Angeles, CA Ketosis prone type 2 diabetes(KPD) has become an increasingly recognized, although incompletely understood entity in the clinical setting over the past few decades. Despite presenting with true anion gap acidosis, these patients show clinical, immunologic, and genetic characteristics classic of type 2 diabetes at follow up. A significant fraction of these patients are able to discontinue insulin therapy after their first episode of DKA. The purpose of our study was to determine which clinical and biochemical characteristics predict beta cell recovery. We performed a retrospective review of patients presenting to the VA Greater LA Medical Center for treatment of DKA between 1999 and 2011. All patients were [gt]18 years old with no prior diagnosis of diabetes. Patients met criteria for DKA upon presentation with plasma glucose [gt]250 mg/dL, ketonemia/moderate ketonuria, and either an anion gap of [gt]13, blood pH [lt]7.3, or bicarbonate level of [lt]20mEq/L. The patients were then divided into those who were able to stop insulin, and those who remained insulin dependent. The groups were compared based on 24 clinical and biochemical parameters. Calculations were made using chi square and Fisher-exact test, along with ANOVA to assess differences in continuous variables. 62 patients met inclusion criteria (3 lost to follow up). A total of 33 patients stopped insulin therapy during the two year follow up period (55%). Our analysis demonstrated no difference between the two populations with respect to individual characteristics. However, patients with 3 or more risk factors (hypertension, dyslipidemia, family history, BMI[gt]25) were more likely to recover, P=0.015. Most recovery patients were autoantibody negative with adequate beta cell function (8/16 patients in whom levels were checked), but this was not predictive of recovery. C-peptide levels at presentation were not predictive of insulin discontinuation, but higher C-peptide levels at [gt]4 weeks correlated with recovery, P=0.017. A1c levels were also significantly lower at presentation in patients who were able to stop insulin, P=0.01. This analysis of 62 ketosis prone diabetic patients revealed that no single clinical characteristic could reliably identify individuals who would become insulin independent long term. Rather, the presence of 3 or more risk factors, robust C-peptide levels at least one month after the initial DKA episode, and lower initial A1c, are all predictive of beta cell recovery in this population.[br][br]Nothing to Disclose: JW, AM, GG 2012-06-24T11:45:00 351 2012-06-24T00:00:00 1899-12-30T11:45:00 3042 155 1022 OR19-3 OR_LB Sunday 849 2012


849 ENDO12L_OR19-4 ORAL SESSION: Late Breaker Oral Session (11:15 AM-12:45 PM) Bisphenol A (BPA), Acute and Chronic Exposure, Alters MiRNA Profiles in Thyroid Cells Elzbieta Iwona Zakrzewska, Julie Dragon, Jeffrey Hayden White, Frances Eileen Carr University of Vermont, Burlington, VT; University of Vermont, Burlington, VT The xenoestrogen BPA, used in the production of polycarbonates, has been shown to disrupt steroid hormone-mediated actions and is linked to the development of prostate and breast cancer. While BPA has been shown to disrupt thyroid hormone-mediated development and associated cellular processes, the effect of BPA exposure on benign and malignant thyroid cells has not yet been studied. Given that miRNA profiles may differ between normal and thyroid cancer tissues corresponding to differential gene expression and that BPA has been shown to alter miRNAs and gene expression in other tissues, we investigated the effect of BPA on miRNA profiles in benign and malignant thyroid cells. Further, we determined the impact of removal of BPA after long-term continuous exposure. Thyroid cells, ThJ (benign) and SW1736 (anaplastic) were maintained in phenol-red free media control (C) or with BPA (10[sup]-6[/sup] M) as follows: (BPA-C) chronic exposure to BPA (3 weeks, 7 passages), (BPA-CR) chronic exposure then subsequent removal of BPA and (BPA-A) acute exposure (1 week, 1 passage). At each passage, cells were collected and total RNA was extracted. RNA was hybridized to miRNA 2.0 chips (Affymetrix, Inc). The reported intensity signals were quantile-normalized, summarized and log2-transformed. Our data indicate that both ThJ and SW1736 cells respond to BPA exposure on the miRNA level. Moreover, BPA alters miRNA expression patterns differently in ThJ and SW1736 cells and the effects vary with acute and chronic BPA exposure. While several of the miRNA changes were reversible as reflected by the log fold change values between BPA-CR and C groups, other miRNAs remained altered even though BPA was no longer present. In benign cells, miR-1274a, miR-1274b, miR-664_s, miR-202 and miR-589 were among those that were up-regulated and miR-196a, miR-200b and c were down-regulated (BPA-C). However, after removal of BPA, miR-589, miR-196a, miR-200b and c did not return to pretreatment levels. In anaplastic cells, miR-155, miR-125b and miR-100 were up-regulated and miR-21* and miR-181c were down-regulated in BPA presence. MiR-100 and miR-181c did not return to pretreatment levels in BPA-CR. These results indicate that acute and chronic exposure to BPA can differentially alter specific miRNA expression in benign and malignant cells. That these changes were not reversed may indicate long-term changes in gene expression that may correspond to permanent changes in cell function and thyroid cancer.[br][br]Sources of Research Support: This publication was funded by College of Medicine, University of Vermont Internal Grant Funding.[br]This publication was made possible by Vermont Genetics Network through Grant Number 8P20GM103449 from the INBRE Program of the National Institute of General Medical Sciences (NIGMS) and the National Center for Research Resources (NCRR), components of the National Institutes of Health (NIH). Its contents are solely the responsibility of the authors and do not necessarily represent the official views of NIGMS or NIH.[br][br]Nothing to Disclose: EIZ, JD, JHW, FEC 2012-06-24T12:00:00 351 2012-06-24T00:00:00 1899-12-30T12:00:00 3052 155 1023 OR19-4 OR_LB Sunday 850 2012


850 ENDO12L_OR19-5 ORAL SESSION: Late Breaker Oral Session (11:15 AM-12:45 PM) Loss-of-Function Mutations in IGSF1 Cause a Novel, X-Linked Syndrome of Central Hypothyroidism and Testicular Enlargement Beata Bak, Yu Sun, Nadia Schoenmakers, Paul AS van Trotsenburg, Wilma Oostdijk, Peter Voshol, Emma Cambridge, Jacqueline K White, Paul le Tissier, Neda M Gharavy, Juan-Pedro Martinez-Barbera, Wilhelmina Stokvis-Brantsma, Thomas Vulsma, Marlies J Kempers, Luca Persani, Irene Campi, Marco Bonomi, Paolo Beck-Peccoz, Hongdong Zhu, Timothy ME Davis, Jayanti J Rangasami, Claudia AL Ruivenkamp, Jeroen FJ Laros, Marjolein Kriek, Sarina G Kant, Cathy AJ Bosch, Nienke R Biermasz, Natasha M Appelman-Dijkstra, Alberto M Pereira, Johan den Dunnen, Martijn H Breuning, Raoul C Hennekam, Krishna Chatterjee, Mehul T Dattani, Daniel J Bernard, Jan M Wit McGill University, Montreal, Canada; Leiden University Medical Center, Leiden, Netherlands; University of Cambridge, Cambridge, UK; Emma Children[apos]s Hospital, Academic Medical Center, Amsterdam, Netherlands; Leiden University Medical Center, Leiden, Netherlands; Wellcome Trust Sanger Institute, Hinxton, UK; National Institute for Medical Research, Mill Hill, UK; UCL Institute of Child Health London, London, UK; Universit[agrave] degli Studi di Milano, Milan, Italy; Universit[agrave] degli Studi di Milano, Milan, Italy; The University of Western Australia, Crawley, Australia; West Middlesex University Hospital, Ilesworth, UK; Leiden University Medical Center, Leiden, Netherlands; UCL Institute of Child Health and Great Ormond Street Hospital for Children, London, UK Congenital hypothyroidism of central origin (CH-C) typically derives from pituitary thyroid-stimulating hormone (TSH) deficiency, either isolated or in conjunction with other hormone deficits. Though considered a rare disorder, the causes of CH-C are poorly characterized. As a result, the condition might be more pervasive than currently appreciated, particularly in milder cases. Undetected CH-C represents a significant clinical problem as untreated hypothyroidism is associated with developmental delay in children and adverse cardiometabolic sequelae in adults. At present, mutations in the thyrotropin-releasing hormone (TRH) receptor (TRHR) or TSH [beta] subunit (TSHB) are the only known genetic causes of isolated TSH deficiency. However, in our clinics, we identified several males with isolated TSH deficiency, testicular enlargement, and variable prolactin deficiency but normal TRHR and TSHB genes. We therefore hypothesized that mutations in an X-linked gene represent a previously unappreciated cause of CH-C. Using whole exome or candidate gene sequencing, we identified nine distinct mutations in the X-linked immunoglobulin superfamily, member 1 (IGSF1) gene in males from 10 families. IGSF1 encodes a pituitary-enriched plasma membrane glycoprotein. Disease-associated mutations blocked trafficking of the IGSF1 protein from the endoplasmic reticulum to the plasma membrane, consistent with the suspected loss of protein function. To definitively link IGSF1 defects to hypothyroidism, we analyzed hypothalamic-pituitary-thyroid function in Igsf1-deficient (Igsf1[Delta]ex1) mice. Adult Igsf1[Delta]ex1 males exhibited markedly decreased pituitary TSH content and circulating thyroxine concentrations, as well as increased body weight and fat mass, recapitulating several features of the human disorder. Trhr, but not Tshb, mRNA levels were significantly reduced in Igsf1[Delta]ex1 pituitaries, suggesting that loss of IGSF1 function might cause CH by impairing TRH signaling. Indeed, patients with IGSF1 mutations secrete TSH with reduced biological activity. Given the role of TRH in prolactin secretion, a reduction in TRHR expression might also underlie prolactin deficiency. The mechanisms of macroorchidism are still under investigation. Collectively, our observations delineate a novel X-linked syndrome of CH-C, testicular enlargement, and variable prolactin deficiency, and reveal a previously unsuspected role for IGSF1 in hypothalamic-pituitary control of thyroid and testicular function.[br][br]Sources of Research Support: NSERC Doctoral Research Award to BB. China Scholarship Council to YS. NSERC Discovery Grant 341801-07; FRSQ Chercheur boursier senior award to DJB. Wellcome Trust grant 095564; National Institutes of Health Research Cambridge Biomedical Research Centre to NS and KC. Wellcome Trust grant WT077157/Z/05/Z to EC and JKW. Wellcome Trust grants 084361, 078432, and 086545 to JPMB. UK MRC U117570590 to PLT. National Health and Medical Research Council of Australia Practitioner Fellowship to TMED. Dutch Growth Research Foundation to DGDB. Young Investigator grant of the Italian Ministry of Health GR-2008-1137632 to MB. Great Ormond Street Children[apos]s Hospital Charity to MTD.[br][br]Nothing to Disclose: BB, YS, NS, PASvT, WO, PV, EC, JKW, PlT, NMG, J-PM-B, WS-B, TV, MJK, LP , IC, MB, PB-P, HZ, TMED, JJR, CALR, JFJL, MK, SGK, CAJB, NRB, NMA-D, AMP, JdD, MHB, RCH, KC, MTD, DJB, JMW 2012-06-24T12:15:00 351 2012-06-24T00:00:00 1899-12-30T12:15:00 3067 155 1024 OR19-5 OR_LB Sunday 851 2012


851 ENDO12L_OR19-6 ORAL SESSION: Late Breaker Oral Session (11:15 AM-12:45 PM) Closed-Loop Insulin Delivery in Children [lt]7 Years of Age [ndash] Identification of Metabolic Parameters Needed To Effect Control Andrew Dauber, Liat Corcia, Sara Einis, Jason Safer, Garry Steil Children[apos]s Hospital Boston, Boston, MA Closed-loop insulin therapy (CLT) shows promise for improving glycemic control in older teenagers and adults; however no CLT studies have been conducted in very young children ([lt]7 years of age). Younger children present a unique challenge for CLT as they have more unpredictable eating and activity patterns than older children, and there is no reason to assume that metabolic parameters describing the rates of glucose appearance following a meal, or the insulin-action time course following subcutaneous insulin delivery, are identical to those observed in the older population. Identification of these parameters is widely believed to be essential for optimizing the algorithms used to calculate closed-loop insulin delivery. We performed the first randomized controlled study (NCT 01421225) comparing CLT with standard pump therapy (SPT) and used the data to identify these metabolic parameters. Children (N=10) were admitted to an inpatient research unit for 48 hours and given identical meals on each day. CLT insulin rates were manually adjusted using sensor data (Abbott Navigator, every 20 minutes form 10 PM to 8 AM and every minute from 8:00 AM to noon). Glycemic control was compared with SPT during the same period (CLT vs. SPT randomized by day). Control was achieved with a physiologic insulin delivery algorithm emulating the B-cell. Subjects ate breakfast at 8 AM with a snack at 10 AM. Glucose levels 4 hours post-breakfasts (pre-lunch) were significantly lower with CLT than SPT (189[plusmn]18 vs. 274[plusmn]23 mg/dL; p=0.0083) but neither therapy achieved the desired daytime target (both different from 120 mg/dL, p[lt]0.05) and the peak post-prandial glucose levels were not different (367[plusmn] 23 vs. 353[plusmn]24 mg/dL, p=0.72 CLT vs SPT). Meal responses were well described by a low order virtual patient model (r[sup]2[/sup]=0.94) characterizing glucose appearance and disposal (one-compartment minimal model). We conclude that CLT can improve postprandial control compared with SPT and that parameters characterizing the meal response can be identified from CLT data, leaving open the possibility to optimize closed-loop control for this age group.[br][br]Sources of Research Support: Abbott Diabetes Care, Animas Corporation, and Hemocue Inc. provided research supplies for this study. This work was conducted with support from Harvard Catalyst | The Harvard Clinical and Translational Science Center (NIH Award #UL1 RR 025758 and financial contributions from Harvard University and its affiliated academic health care centers). The content is solely the responsibility of the authors and does not necessarily represent the official views of Harvard Catalyst, Harvard University and its affiliated academic health care centers, the National Center for Research Resources, or the National Institutes of Health.[br][br]Nothing to Disclose: AD, LC, SE, JS, GS 2012-06-24T12:30:00 351 2012-06-24T00:00:00 1899-12-30T12:30:00 3078 155 1025 OR19-6 OR_LB Sunday 852 2012


852 ENDO12L_SUN-1 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Granulosa-Cell Specific Deletion of Notch2 Results in Aberrant Ovarian Follicle Formation Dallas A Vanorny, Signe M Kilen, Kelly E Mayo Northwestern University, Evanston, IL; Northwestern University, Evanston, IL During ovarian development, primordial follicles form through a process known as germ cell nest breakdown, where germ cells in syncytia (nests) are encapsulated by somatic pre-granulosa cells. Using a transgenic mouse line containing a Notch-responsive GFP reporter (TNR line provided by N. Gaiano, JHU, Baltimore), we visualized Notch activity in the somatic pre-granulosa cells that surround germ cells at the time of nest breakdown, and demonstrated that this signal was suppressed by gamma-secretase inhibitors that block Notch signaling. These somatic cells are known to express the receptor Notch2, and to more fully explore the involvement of Notch signaling in nest breakdown, we used the Cre/loxP system to disrupt the Notch2 gene specifically within granulosa cells (floxed mice provided by U. Strobl, LMU, Munich; Amhr2[sup]Cre[/sup] transgenic mice provided by R. Behringer, M.D. Anderson, Houston). Notch2 knockout mice (Amhr2[sup]Cre/+[/sup],Notch2[sup]fl/-[/sup] or N2KO) have a clear decrease in ovarian Notch2 protein levels as detected by immunohistochemistry compared to controls (Amhr2[sup]Cre/+[/sup],Notch2[sup]+/-[/sup] or N2het and wild-type (WT)). However, the level of Notch2 varies considerably among N2KO mice (reduction of 40-100% compared to WT by Western blot). Examination of N2KO ovaries revealed the presence of numerous multi-oocytic follicles (MOFs) containing more than one oocyte, some with as many as 13 oocytes. N2KO ovaries also contain follicles with enlarged or degenerating oocytes and exhibit an increase in granulosa cell death as determined by TUNEL staining. Despite these abnormalities, female N2KO mice are fertile, with a mean litter size similar to WT. However, several of the N2KO females examined exhibited premature reproductive senescence. These findings indicate an important role for Notch2 in the mouse ovary, and together with our earlier studies examining germ cell specific disruption of the Notch ligand Jagged1, they support a critical role for the Notch pathway in germ cell nest breakdown and primordial follicle formation. Current studies are aimed at understanding how loss of Notch2 impacts granulosa cell function and follicle development.[br][br]Sources of Research Support: NIH P01 HD021921 and NIH T32 GM12453.[br][br]Nothing to Disclose: DAV, SMK, KEM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1919 173 1059 SUN-1 PO33-01 Sunday 853 2012


853 ENDO12L_SUN-2 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Coordinated Response of Ovarian Follicles to Endocrine and Paracrine Signals Via Controlled Heparan Sulfate Proteoglycan Synthesis Laura N Watson, Rebecca L Robker, Kylie R Dunning, David G Mottershead, Robert B Gilchrist, Darryl L Russell University of Adelaide, Adelaide, Australia In the ovarian follicle mural granulosa and cumulus cells perform key specific roles in endocrine communication and oocyte maturation. Oocyte secreted growth factors promote cumulus and repress mural granulosa cell specific genes to establish these two cell lineages and regulate cumulus cell function. The mechanism establishing this precise morphogenic gradient of oocyte signalling within the follicle is unknown. The present study investigated a role for heparan sulphate proteoglycans as co-receptors mediating oocyte derived growth factor signalling. Heparin treatment of in vitro maturation cultures of cumulus oocyte complexes (COC) prevented cumulus expansion, blocked the induction of cumulus specific matrix genes [italic]Has2[/italic] and [italic]Tnfaip6,[/italic] while conversely strongly upregulating mural granulosa specific genes [italic]Lhcgr[/italic] and [italic]Cyp11a1[/italic]. Heparin also blocked phosphorylation of the GDF9 signalling mediator Smad2. However, when heparin treated cultures were co-treated with exogenous GDF9, all of the effects of heparin were completely reversed. Furthermore, recombinant pro-GDF9 strongly bound heparin sepharose. These observations indicate that heparin disrupts interaction of endogenous GDF9 with a heparin sulphate proteoglycan co-receptor required for GDF9 signalling. The expression of all cell surface HSPG from the syndecan and glypican families as well as betaglycan and CD44 are under dynamic endocrine control. In preovulatory follicles expression of all the glypicans were more abundant in mural granulosa than cumulus cells while syndecans and betaglycan abundance was similar. An ovulatory dose of hCG caused downregulation of betaglycan in cumulus cells and this regulation required GDF9 activity, conversely betaglycan was significantly increased in luteinising mural granulosa cells. hCG caused very strong induction of Syndecan 1 and 4 in mural granulosa as well as cumulus cells. Glypican 1 was selectively induced in cumulus cells and this expression was dependent on GDF9 action. Together the data suggest that HSPG play an essential role in GDF9 signalling and, based on cell- and temporal-specific regulation, implicates glypican 1 as a key GDF9 co-receptor involved in patterning of oocyte signalling and cumulus cell function in the periovulatory follicle.[br][br]Sources of Research Support: National Health and Medical Research Counil Australia. Australian Research Council.[br][br]Nothing to Disclose: LNW, RLR, KRD, DGM, RBG, DLR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1503 173 1060 SUN-2 PO33-01 Sunday 854 2012


854 ENDO12L_SUN-3 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) An Androgen-Induced microRNA Regulates Androgen Actions in Follicular Development and Function Aritro Sen, Stephen R Hammes University of Rochester Medical Center, Rochester, NY Androgens have traditionally been considered detrimental to women[apos]s health and associated with deregulated ovarian function and infertility. Interestingly, through the generation of a granulosa cell (GC)-specific androgen receptor (AR) knockout (ARKO) mouse model, we have recently pioneered a new concept that critical androgen actions in GCs are absolutely essential for normal ovarian function and female fertility. Our studies indicate that ARs expressed in GCs control pre-antral follicle growth and development to antral follicles, while preventing follicular atresia. However, to date, the underlying mechanism of this AR action has remained elusive. We have now uncovered a novel regulatory pathway in GCs whereby ARs regulate the expression of a micro-RNA (miR) that may contribute to androgen-induced follicular survival. The 5[apos] UTR region of this miR contains androgen responsive elements, suggesting its expression might be dependent on AR-mediated transcription. Interestingly, expression of this miR is also regulated by Erk1/2. It is well established that androgen functions are mediated by both [ldquo]genomic[rdquo] and [ldquo]membrane-initiated[rdquo] actions of ARs. Furthermore, we have found that androgens can promote MAPK signaling in a mechanism conserved from Xenopus to mouse and humans. Accordingly, we find that expression of this miR in GCs requires a synergistic action between the nuclear and extranuclear actions of ARs. Intriguingly, down-regulation of this miR by siRNA in primary mouse GC cultures leads to increased expression of pro-apoptotic proteins. Furthermore, we found that, in GC-specific ARKO mice, expression of the miR is significantly lower, while expression of pro-apoptotic protein levels (and caspase 3 activity) are considerably higher compared to wild type animals. Together, our results suggest that genomic and membrane-initiated androgen signaling induces expression of this miR, which then leads to the suppression of pro-apoptotic protein expression in GCs, decreased follicular atresia, and follicular survival. Thus, androgen-induced miR may account at least in part for androgen actions in follicular development and function. Ongoing studies are aimed towards determining the role of this miR in mediating androgen actions during both normal and pathologic follicular development in vivo.[br][br]Nothing to Disclose: AS, SRH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 437 173 1061 SUN-3 PO33-01 Sunday 855 2012


855 ENDO12L_SUN-4 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Enhanced Survival of Mouse Ovarian Follicles Cultured in Low Oxygen Tension Yogeshwar Makanji, Jennifer E Pahnke, Teresa K Woodruff Feinberg School of Medicine, Northwestern University, Chicago, IL Folliculogenesis is a complex process controlled by various factors. Unraveling this process requires a better comprehension of the physiology of follicle growth initiation, the interplay of important growth factors, the basic metabolic needs of early growing follicles and the physical environment of the follicle. A proposed activation signal for early follicles is oxygen. Studying oxygen needs in a multi-cellular organ such as the follicle, which changes somatic cell numbers at a rapid rate, will increase our knowledge about ovarian follicle maturation. Recently, a three-dimensional alginate system for follicle maturation has been developed in our laboratory and represents a novel method of replicating the in vivo environment of the growing follicle (1). In this study, late primary follicles were cultured in low oxygen tension to determine the effect of O[sub]2[/sub] on growth and survival of mouse ovarian follicles.[br]Ovarian follicles (85- 130 [mu]m) were isolated from female CD1 mice and encapsulated in 0.25% alginate. The follicles enclosed within an alginate bead were cultured in 200 [mu]l of growth media ([alpha]MEM, 3 mg/ml bSA, 1 mg/ml bovine fetuin, 10 mIU/ml FSH, 5 [mu]g/ml insulin, 5 [mu]g/ml transferrin and 5 ng/ml selenite pH 7.4) at 2.5, 5 or 20% O[sub]2[/sub], 5% CO[sub]2[/sub], 37[deg]C for 8 days. At the end of culture the size of the follicle was measured and analyzed using Image J software. The culture media was assayed for mouse VEGF and estradiol in ELISAs.[br]Follicles that had a starting size of 100 [mu]m had a 80% survival rate when cultured in 2.5 or 5% O2 compared to 20% survival when cultured in 20% O[sub]2[/sub]. In addition, growth was improved at lower O[sub]2[/sub] tension, follicles cultured in 2.5% O[sub]2[/sub] with a starting size of 108 [plusmn] 10 [mu]m grew to 180 [plusmn] 33 [mu]m (67% growth) in 8 days, a similar growth rate was observed for follicles grown at 5% O[sub]2[/sub]. In comparison, follicles cultured in 20% O[sub]2[/sub] with a starting size of 106 [plusmn] 6 [mu]m grew to 135 [plusmn] 13 [mu]m (28% growth). Interestingly, the survival of follicles was directly correlated to VEGF levels, follicles cultured in 2.5% O[sub]2[/sub] had 128 pg/ml, 5% O[sub]2[/sub] had 59 pg/ml and 20% O[sub]2[/sub] had undetectable levels. In addition, estradiol levels were correlated to follicle size/stage.[br]In conclusion, low oxygen tension (2.5 and 5% O[sub]2[/sub]) improves survival of late primary mouse follicles and these follicles secrete more VEGF compared to ones cultured in high O[sub]2[/sub] tension (20% O[sub]2[/sub]).[br][br]1. Xu, M. et al., Tissue Engineering 2006; 12:2739.[br][br]Sources of Research Support: This work was supported by NIH funding to the U54 Center for Reproductive Research, Northwestern University (HD04187) and a NHMRC Overseas Biomedical Training Fellowship (GNT1016460) to Y.M.[br][br]Nothing to Disclose: YM, JEP, TKW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1916 173 1062 SUN-4 PO33-01 Sunday 856 2012


856 ENDO12L_SUN-5 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Encapsulated Ovarian Endocrine Cells Produce Sustained Levels of Steroid Hormones [italic]In Vivo[/italic] Sittadjody Sivanandane, Justin M Saul, Sunyoung Joo, Rajesh Pareta, Juan Arenas, Nathaniel T Marshall, James J Yoo, Anthony Atala, Emmanuel C Opara Wake Forest University School of Medicine, Winston-Salem, NC; Miami University, Oxford, OH Reduced sex steroid production from the ovaries due to surgical resection, ablative therapy, or menopause leads to various physiological consequences in women (1). Although hormone replacement therapy is able to compensate for the loss of hormone production, delivery through pharmacological means results in consistently high serum hormone concentrations and clinical complications including increased incidence of heart disease and cancer (2). Cell-based hormone delivery offers the potential to overcome these complications because it provides a system that is capable of responding to physiological stimuli such as serum hormone levels. To date, however, such a cell-based approach has not been reported. Therefore, the purpose of our study was to design a tissue-engineered construct of the endocrine ovary using encapsulation techniques and to test the function of the construct [italic]in vivo[/italic]. Ovaries were excised from 21 day old Fisher 344 rats, and the endocrine cells (granulosa and theca cells). Granulosa and theca cells were then encapsulated in alginate hydrogel microcapsules to provide three dimensional support. The granulosa cells were positioned in a central alginate core, and theca cells were placed in an outer alginate layer of multi-layer microcapsules, a structure similar to the arrangement of these cells in the native follicles. The two alginate layers were separated by a semi-permeable membrane made of poly-L-Ornithine. The ovarian tissue constructs were implanted in omentum pouches created in each of 5 ovariectomized (ovx) Fisher 344 rats. A Control group of 5 ovx rats were implanted with blank microcapsules in their omentum pouches. Blood samples were collected periodically from all animals for 4 weeks, and plasma levels of 17 b-estradiol (E[sub]2[/sub]) progesterone (P[sub]4[/sub]), follicle-stimulating hormone (FSH) and Luteinizing hormone (LH) were measured by ELISA. The data were evaluated by one-way ANOVA using SPSS. While the plasma levels of E[sub]2[/sub] and P[sub]4[/sub] in ovx rats implanted with the tissue construct were significantly higher, FSH and LH levels were significantly lower than those measured in the control group throughout the follow up. The encapsulation techniques, demonstrate for the first time that the endocrine unit of the ovary could be recapitulated [italic]ex vivo a[/italic]nd the tissue-engineered construct produced sustained levels of E[sub]2[/sub] and P[sub]4[/sub] [italic]in vivo, [/italic]which regulated the levels of gonadotropins through negative feedback mechanism.[br][br]1. Stevenson JC. A woman[apos]s journey through the reproductive, transitional and postmenopausal periods of life: Impact on cardiovascular and musculo-skeletal risk and the role of estrogen replacement. Maturitas (2011); 70: 197-205. 2. Beral V, Banks E, Reeves G, Appleby P. Use of HRT and the subsequent risk of cancer. J Epidemiol. Biostat. (1999); 4(3):191-215.[br][br]Sources of Research Support: This study was supported by funding from Jack and Pamela Egan.[br][br]Nothing to Disclose: SS, JMS, SJ, RP, JA, NTM, JJY, AA, ECO 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2125 173 1063 SUN-5 PO33-01 Sunday 857 2012


857 ENDO12L_SUN-7 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Identification of a Novel Ovarian Specific Isoform Promoter of Liver Receptor Homolog-1 in Ovarian Granulosa Cells Shinya Kawabe, Takashi Yazawa, Masafumi Kanno, Yoko Usami, Tetsuya Mizutani, Yoshitaka Imamichi, Yunfeng Ju, Takahiro Matsumura, Kaoru Miyamoto University of Fukui, Fukui, Japan; University of Fukui, Fukui, Japan; University of Fukui, Fukui, Japan Liver receptor homolog-1 (LRH-1) belongs to NR5A nuclear receptor (NR) family. LRH-1 plays important roles not only in endodermal tissues but in mesodermal tissues, especially in the ovary. In the ovary, LRH-1 is expressed in granulosa cells and known to be involved in regulation of genes related to steroidogenesis and ovulation. An LRH-1 promoter working in the liver was not functional in ovarian granulosa cells. To reveal transcriptional regulatory mechanism of LRH-1 in granulosa cells, we determined the transcription start site (TSS) of LRH-1 in the ovary using KGN cells, a human granulosa cell tumor cell line. 5[prime]-RACE PCR revealed that the TSS of LRH-1 located at 41 bp upstream of the reported exon 2 in KGN cells. The novel isoform of LRH-1 was expressed in the human ovary but not in the liver. For luciferase reporter assays, we constructed reporter plasmids containing upstream region of the new exon 1. Deletion and mutational analysis revealed that two sites, a steroidogenic factor-1 (SF-1) binding site and a GC box, were required for the reporter activity in rat granulosa cells as well as in KGN cells. Electrophoretic mobility shift assay showed that SF-1 and specificity protein (Sp)-1/3 bind to the respective regions. Co-transfection of SF-1 increased promoter activity of LRH-1 in KGN cells. Interestingly, the SF-1 dependent reporter activity was further enhanced when peroxisome proliferator-activated receptor-[gamma] coactivator-1[alpha] (PGC-1[alpha]) is co-transfected. In [italic]Drosophila[/italic] SL2 cells lacking endogenous Sp1 and Sp3, Sp1 was more effective than Sp3 for the promoter activity, and co-transfection of SF-1 synergistically increased the activity. Infection with adenoviruses expressing SF-1 or PGC-1[alpha] induced expression of LRH-1 in KGN cells. These results indicate that the expression of LRH-1 is regulated in a tissue-specific manner, and that the novel promoter region is coordinately controlled by SF-1, PGC-1[alpha] and Sp1/3 in ovarian granulosa cells.[br][br]Nothing to Disclose: SK, TY, MK, YU, TM, YI, YJ, TM, KM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 874 173 1064 SUN-7 PO33-01 Sunday 859 2012


858 ENDO12L_SUN-8 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Roles of Cyp26b1 and Retinoic Acid in the Proliferation and Apoptosis of Normal Ovarian Granulosa Cells and Ovarian Cancer Cells Jingjing Kipp, Michael Demczuk, Ann Golebiowski, Kelly Mayo, Brendan Butkut DePaul University, Chicago, IL; Northwestern University, Evanston, IL Cyp26b1, a member of the cytochrome P450 family, encodes an enzyme that degrades retinoic acid (RA). We recently demonstrated expression of Cyp26b1 in postnatal ovarian granulosa cells and discovered that Cyp26b1 gene expression is strongly inhibited by activin. We found that activin, RA and the Cyp26 inhibitor R115866 increased granulosa cell numbers. In addition to granulosa cells, we also localized Cyp26b1 mRNA in ovarian surface epithelial cells. This study was designed to examine expression of Cyp26b1 in a variety of ovarian cancer cell lines and further investigate the consequences of Cyp26b1 overexpression, silencing, or treatment with a Cyp26 inhibitor or RA on the proliferation and apoptosis of normal granulosa cells and ovarian cancer cells. The granulosa tumor cells KGN and the epithelial ovarian cancer cells SKOV-3, OVCAR-3 and Caov-4 all showed high levels of Cyp26b1 expression. Cyp26b1 mRNA levels in KGN cells were elevated as compared to normal human granulosa-lutein cells (collected at IVF), suggesting an association between overexpression of this gene and the ovarian cancer phenotype. Consistent with the suppressive effect of activin on Cyp26b1 expression, activin [beta]A subunit mRNA levels in the three different granulosa cell types are inversely correlated with Cyp26b1 mRNA levels, and activin [beta]A mRNA levels were low in the epithelial ovarian cancer cell lines. Treatment with R115866 suppressed cell proliferation in KGN, SKOV-3 and Caov-4 cells, indicating that increased Cyp26b1 expression is associated with ovarian cancer cell proliferation and decreased Cyp26b1 levels can lead to suppression of growth. This observation was further confirmed by increased cell apoptosis in SKOV-3 cells transfected with siRNAs targeting Cyp26b1. Activin A also suppressed proliferation of these three ovarian cancer cell lines, consistent with a recent report and with our finding that activin suppresses Cyp26b1 expression. In primary cultured normal mouse granulosa cells, overexpression of Cyp26b1 suppressed granulosa cell proliferation and induced cell apoptosis, while R115866 and RA stimulated cell proliferation and suppressed apoptosis. Overall, this study provides evidence that Cyp26b1 inhibits, while RA promotes, normal granulosa cell growth through regulation of proliferation and apoptosis, and that Cyp26b1 has an opposite effect on ovarian cancer cell proliferation and apoptosis.[br][br]Sources of Research Support: DePaul University Research Council Grant (JK).[br][br]Nothing to Disclose: JK, MD, AG, KM, BB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2124 173 1065 SUN-8 PO33-01 Sunday 860 2012


859 ENDO12L_SUN-9 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Resveratrol Promotes Expression of SIRT1 and StAR in Rat Ovarian Granulosa Cells; Possible Implication of SIRT1 in the Ovary Osamu Hiraike, Yoshihiro Morita, Tetsu Yano, Akira Shirane, Mana Hirano, Haruko Hiraike, Osamu Yoshino, Yuichiro Miyamoto, Katsutoshi Oda, Kei Kawana, Shunsuke Nakagawa, Kazuyoshi Tsutsui, Shiro Kozuma, Yuji Taketani The University of Tokyo, Tokyo, Japan; Teikyo University, Tokyo, Japan; Waseda University, Tokyo, Japan Background; Resveratrol is a natural polyphenolic compound known for its beneficial effects on energy homeostasis, and it also has multiple properties, including anti-oxidant, anti-inflammatory, and anti-tumor activities. Recently, silent information regulator genes (Sirtuins) have been identified as targets of resveratrol. Sirtuin 1 (SIRT1), originally found as an NAD+-dependent histone deacetylase, is a principal modulator of pathways downstream of calorie restriction, and the activation of SIRT1 ameliorates glucose homeostasis and insulin sensitivity. To date, the presence and physiological role of SIRT1 in the ovary are not known. Here we found that SIRT1 was localized in granulosa cells of the human ovary.[br]Methods; The physiological roles of resveratrol and SIRT1 in the ovary were analyzed. Immunohistochemistry was performed to localize the SIRT1 expression. SIRT1 protein expression of cultured cells and luteinized human granulosa cells was investigated by Western blot. Rat granulosa cells were obtained from diethylstilbestrol treated rats. The cells were treated with increasing doses of resveratrol, and subsequently harvested to determine mRNA levels and protein levels. Cell viability was tested by MTS assay. Cellular apoptosis was analyzed by caspase 3/7 activity test and Hoechst 33342 staining.[br]Results; SIRT1 protein was expressed in the human ovarian tissues and human luteinized granulosa cells. We demonstrated that resveratrol exhibited a potent concentration-dependent inhibition of rat granulosa cells viability. However, resveratrol-induced inhibition of rat granulosa cells viability is independent of apoptosis signal. Resveratrol increased mRNA levels of SIRT1, LH receptor, StAR, and P450 aromatase, while mRNA levels of FSH receptor remained unchanged. Western blot analysis was consistent with the results of quantitative real-time RT-PCR assay. In addition, progesterone secretion was induced by the treatment of resveratrol.[br]Conclusions; These results suggest a novel mechanism that resveratrol could enhance progesterone secretion and expression of luteinization-related genes in the ovary, and thus provide important implications to understand the mechanism of luteal phase deficiency.[br][br]Nothing to Disclose: OH, YM, TY, AS, MH, HH, OY, YM, KO, KK, SN, KT, SK, YT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 137 173 1066 SUN-9 PO33-01 Sunday 861 2012


860 ENDO12L_SUN-10 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Synergistic Effect of RAGE and AR Pathways in PCOS Deyarina Gonzalez, Armando Rojas, Amy K White, Jacqueline Romero, Julia Davies, Rafael Gracia, Claudia Velez, Lisa Joels, John O White, Robert Steven Conlan Swansea University, Swansea, UK; Catholic University of Maule, Talca, Chile; Royal Devon and Exeter Hospital, Devon, UK The endometrium of Polycystic ovary syndrome (PCOS) women is characterised by elevated expression and enhanced transcriptional activity of androgen receptor (AR) and altered expression of implantation markers such as mucin 1 (MUC1). In PCOS patients elevated levels of circulating Advanced Glycation end-products (AGEs) correlate with increased testosterone levels suggesting a possible interaction between AGE and hyperandrogenaemia. Objectives: To determine 1) whether the receptor for AGE (RAGE) is expressed in endometrium 2) the association between endometrial expression of RAGE and circulating androgen levels in fertile and infertile PCOS women 3) AGE-RAGE effect on the transcriptional activity of AR. Patients and Methods: 30 fertile and 40 infertile PCOS women were recruited for this study. Blood samples were collected at day 2 of the menstrual cycle and serum androgens (testosterone, SHBG, 17-hydroxy- DHEA-S) measured. An endometrial biopsy was collected at LH+7 from each patient and RAGE expression was determined by immunohistochemistry, RT-PCR and Western blot. AGE/RAGE signalling and AR transcriptional activity on the RAGE and MUC1 promoter was assessed by ChIP. Results: Fertile endometrium expresses low amounts of RAGE protein and transcript. RAGE expression in the glandular (2.5 fold, p=0.016), luminal (2 fold, 0.026) and stromal (2.2.5 fold p=0.045) compartments was stronger in PCOS endometrium compared to fertile. In PCOS patients elevated RAGE protein levels markedly correlated to free androgens levels (p=0.00670). Higher endometrial levels of RAGE transcript were observed in PCOS samples compared to fertiles (p=0.023). Incubation of PCOS and fertile endometrial cells with AGEs, DHT or in combination increased the expression of RAGE and MUC1 whereas casodex inhibited this effect. AR occupancy of the RAGE promoter was higher in PCOS samples and it was mediated by AP1 and SP1 proteins. AGE stimulation increased the occupancy of AR on RAGE promoter. AGE/RAGE effect on MUC1 expression was mediated by enhanced occupancy of p65 on MUC1 promoter. Conclusions: RAGE signalling pathway is active in endometrium and is involved in the expression of AR and NFkB target genes. The AR pathway regulates RAGE endometrial expression. A synergistic effect between AGE/RAGE and AR pathway occurs in endometrium. Enhanced AGE/RAGE signalling in PCOS endometrium may alter endometrial function and contribute towards PCOS infertility.[br][br]Nothing to Disclose: DG, AR, AKW, JR, JD, RG, CV, LJ, JOW, RSC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 683 173 1067 SUN-10 PO33-01 Sunday 862 2012


861 ENDO12L_SUN-11 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) MUC4 Regulation by Cytokines and Steroid Hormones in Human Uterine Epithelial Cell Lines Patricia Jensen Chapela, Daniel Carson Rice University, Houston, TX Transmembrane mucins are very large glycoproteins that lubricate and protect the surface of epithelial cells from pathogens and environmental challenges. Their abundance and physical size sterically hinders access to many cell surface receptors. In the context of embryo implantation, these mucins represent a barrier that must be removed to permit embryo attachment at the apical surface of uterine epithelial cells. MUC4 is one transmembrane mucin produced by uterine epithelia. We have used quantitative real time PCR (qRT-PCR) to profile MUC4 expression in a variety of human uterine epithelial cell lines, including many of tumor origin. Previous studies from our lab demonstrated that progesterone and the proinflammatory cytokines, interferon-[gamma] (IFN[gamma]) and tumor necrosis factor [alpha] (TNF[alpha]) stimulate expression of another transmembrane mucin, MUC1, in various contexts (1,2). We have tested the hypothesis that these same agents also stimulate MUC4 expression in two uterine epithelial cell lines, IHEEC (derived from normal endometrium) and HEC50 (derived from a uterine adenocarcinoma) using qRT-PCR. Neither progesterone nor estrogen altered the levels of MUC4 mRNA in either cell line, even though IHEECs were shown to be responsive to both hormones in transient transfection assays using a reporter driven by consensus steroid hormone response elements. In contrast, interferon-[gamma] (IFN[gamma]) and tumor necrosis factor-[alpha] (TNF[alpha]) alone increased MUC4 mRNA levels up to 6-9-fold in the IHEEC cell line in a dose-dependent fashion. In combination, these same cytokines displayed a large synergistic stimulation of [gt]60-fold. In HEC50 cells, IFN[gamma] increased MUC4 mRNA levels 3-fold. No response was observed after treatment with TNF[alpha]; however, a synergistic 9-fold increase in mRNA was observed when treated with TNF[alpha] in combination with IFN[gamma]. The effects of these cytokines on MUC4 protein expression and promoter activity are currently being examined. Collectively, these observations indicate that MUC4 is a sensitive target of proinflammatory cytokine action in both normal and tumor-derived uterine epithelia.[br][br](1) Brayman et al., 2006. Mol. Endocrinol. 20:2278. (2) Dharmaraj et al. 2010. Mol. Endocrinol. 24: 2253.[br][br]Sources of Research Support: NIH grant HD29963 to DDC.[br][br]Nothing to Disclose: PJC, DC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2255 173 1068 SUN-11 PO33-01 Sunday 863 2012


862 ENDO12L_SUN-12 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) MUC16 Stimulation by Proinflammatory Cytokines in the Immortalized Human Endometrial Epithelial Cell Line, IHEEC Micaela Morgado, Neeraja Dharmaraj, Daniel Carson Rice University, Houston, TX; MD Anderson, Houston, TX MUC16 is a member of the family of high molecular weight, heavily glycosylated transmembrane mucin glycoproteins and is primarily found at the apical surface of normal simple epithelia. In general, mucins lubricate and protect epithelial cell surfaces from pathogens and environmental challenges. Transmembrane mucins also influence normal uterine function by acting as barriers that must be removed to allow embryo attachment. MUC16, also known as CA 125, also binds mesothelin and is likely to contribute to abnormal cell colonization of mesothelial surfaces in cancer and endometriosis. Previous studies have shown that MUC1 is highly regulated by the proinflammatory cytokines, tumor necrosis factor [alpha] (TNF[alpha]) and interferon [gamma] (IFN[gamma]), as well as by progesterone in uterine epithelia and other cellular contexts. All three factors are detected at high levels in uterine tissues at the same time that MUC1 is highly expressed. We hypothesized that these same factors also stimulate MUC16 expression in the complex uterine environment. Herein, we report that MUC16 mRNA expression is stimulated 2-3-fold by TNF[alpha] (2.5ng/ml) and 3-4-fold IFN[gamma] (20 IU/ml) in IHEEC cells, a telomerase immortalized human endometrial epithelial cell line derived from normal endometrium. Interestingly, combined treatment with both cytokines resulted in a large (20-60 fold), synergistic stimulation of MUC16 mRNA levels. In contrast, progesterone (200 nM) had a modest response (2 fold) on MUC16 mRNA levels. Currently, we are examining cytokine control of MUC16 promoter activity and antagonism of cytokine stimulation by thiazolidinediones, known to inhibit MUC1 expression in a variety of contexts.[br][br]Sources of Research Support: NIH grant HD29963 awarded to DDC.[br][br]Nothing to Disclose: MM, ND, DC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1835 173 1069 SUN-12 PO33-01 Sunday 864 2012


863 ENDO12L_SUN-13 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Regulation of SIRT1 Determines Initial Step of Endometrial Receptivity by Controlling E-Cadherin Expression Akira Shirane, Osamu Hiraike, Mana Hirano, Michihiro Tanikawa, Yoshihiro Morita, Takayuki Seiki, Haruko Hiraike, Yuichiro Miyamoto, Kenbun Sone, Hajime Oishi, Katsutoshi Oda, Kei Kawana, Shunsuke Nakagawa, Tetsu Yano, Shiro Kozuma, Yuji Taketani The University of Tokyo, Tokyo, Japan; Teikyo University School of Medicine, Tokyo, Japan Background[br]Sirtuin 1 (SIRT1), originally found as a class III histone deacetylase, is a principal modulator of pathways downstream of calorie restriction, and the activation of SIRT1 ameliorates glucose homeostasis and insulin sensitivity. To date, physiological roles of SIRT1 in human uterine tissue remain elusive.[br]Methods[br]We examined whether the expression of SIRT1 regulates E-cadherin expression by Western blot and luciferase assay. Chemical compounds that positively regulates SIRT1 (resveratrol and AICAR) and negatively regulates SIRT1 (sirtinol and nicotinamide) were investigated to affect the expression of E-cadherin.[br]Result[br]Exogenous expression of SIRT1 significantly enhanced E-cadherin expression, while small interfering RNA-mediated depletion of endogenous SIRT1 resulted in a significant reduction of E-cadherin expression. Resveratrol elevated E-cadherin expression in a dose dependent manner, while Sirtinol and nicotinamide exhibited a dose dependent reduction of E-cadherin expression. We also showed that both expression of SIRT1 and activation of SIRT1 promote E-cadherin-driven reporter gene constructs.[br]Conclusion[br]The initial step of human reproduction depends on the capacity of an embryo to attach and implant into the endometrial wall, and these results revealed the novel mechanism that activation and increased expression of SIRT1 play an important role in uterine receptivity.[br][br]Nothing to Disclose: AS, OH, MH, MT, YM, TS, HH, YM, KS, HO, KO, KK, SN, TY, SK , YT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 355 173 1070 SUN-13 PO33-01 Sunday 865 2012


864 ENDO12L_SUN-14 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Nanomechanical Properties of the Endometrial Epithelial Surface Directly Correlated with Characteristic Mucin Expression Patterns Lewis Webb Francis, Sean Geraint Griffiths, John Owen White, Alexandre Berquand, Robert Steven Conlan Swansea University, Swansea, UK; Bruker Nano Services, Manheim, Germany Atomic force microscopy offers a unique contact microscopy method to functionally analyse the role of the signature molecular patterns of the endometrial epithelial cell surface. Coupled with detailed patterning of the protein mosaic network at the cell surface, AFM has the ability to fully delineate the roles of the functional endometrium leading to targeted therapeutics for receptivity and fertility. The window of implantation, between days 21 and 25 of the menstrual cycle, has been heavily characterised by molecular biology and the apical layer presented to the blastocyst is hypothesised to be functionally adhesive and heterogenous. Objectives: To determine (1) the Mucin protein expression in an in vitro model of receptive endometrium and correlate with expression patterns of fertile endometrium (2) to characterise the nanomechanical and adhesion properties of the cells at each stage of the in vitro cycle (3) to directly correlate MUC1 expression with associated functional biophysical characteristics of the endometrial surface. Methods: An in vitro model for endometrial receptivity was simulated using physiologically relevant treatments of endometrial epithelial cell lines Hec-1-A and Hec-1-B with estradiol, progesterone or combinations of both, or with siRNA targeting MUC1. MUC1 protein expression and localisation in the monolayer was assessed using the IN Cell protein analyser (GE healthcare) and correlated with nanomechanical functional characterisation and single molecule force spectroscopy. Results: Hec-1-A and Hec-1-B exhibit differing levels of basal MUC1 expression at the RNA and protein level. Altered effects following estradiol and progesterone treatment were also observed, in line with the presence or absence of the requisite hormone receptors ER and PR respectively. AFM was uniquely able to delineate the functional consequence of altered MUC1 expression patterns and enabled the hypothesis that MUC1 is indeed an anti adhesive protein during the window of implantation to be demonstrated. MUC1 siRNA significantly lowered the protein expression level of MUC1 and altered its monolayer distribution pattern. AFM nano-indentation and single molecule force characterisation of cellular properties, through morphological and nanomechanical observation, has for the first time allowed the direct correlation of molecular expression patterns and functional adhesion properties in endometrial epithelial monolayers.[br][br]Nothing to Disclose: LWF, SGG, JOW, AB, RSC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1566 173 1071 SUN-14 PO33-01 Sunday 866 2012


865 ENDO12L_SUN-15 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Annexin A5 of Maternal Blood Is Prerequisite for Preventing Fetal Loss during Intact Pregnancy Mitsumori Kawaminami, Hiroshi Ueki, Yasuhiro Nishimura, Duangjai Rieanrakwong, Tomohiro Yonezawa, Shiro Kurusu, Yoshihisa Hasegawa, Bent Brachvogel, Ernst Poschl Kitasato University, Towada, Japan; Kitasato University, Towada, Japan; University of Cologne, Cologne, Germany; University of East Anglia, Norwich, UK Antiphospholipid syndrome is characterised by the presence of autoantibodies against phospholipids and/or phospholipid-binding proteins. It is associated with an increased risk of thrombosis and pregnancy loss. Annexin A5, a proposed placental anticoagulant protein, is a candidate antigen in antiphospholipid syndrome. However, it has not been known whether endogenous annexin A5 prevents fetal loss during normal pregnancy. We found, in the present study, that annexin A5-deficient (Anxa5-KO) mice demonstrate a significant reduction in litter size due to sporadic growth retardation of embryo and fetal loss occurring in Anxa5-KO mothers. There was no difference in estrous cyclicity and ovulation between Anxa5-KO and control C57BL/6J. Significant fetal loss occurred between pregnancy day 12 and 18. No further reduction of fetus was observed after day 18. Smaller fetuses were found in the uterus of Anxa5-KO mice on days 12, 15 and 18 of pregnancy, indicating that some embryos were restricted in growth and resorbed during late gestation. Large fibrin-like clots were always observed both in the labyrinth and junctional zone placenta of smaller fetuses. In cross-breeding experiments of Anxa5-KO and C57BL/6J, only litters from Anxa5-KO females showed a reduction in number. Annexin A5 was demonstrated in maternal wild-type mice to cover all surfaces of the central arterial canal in the spongiotrophoblast region and syncytiotrophoblasts of the labyrinth zone that maternal blood bathed, whereas this annexin A5 was absent when mother was Anxa5-KO. Blood isolated from Anxa5-KO mice coagulated significantly faster than that of C57BL/6J. Finally, subcutaneous administration of the anticoagulant heparin (100 IU/day on pregnancy days 12, 14 and 16) to Anxa5-KO mothers significantly reduced pregnancy loss. Hence, thrombosis in the placenta and fetal loss are caused by the absence of maternal annexin A5. These results provide direct evidence that maternal annexin A5 works as an anti-coagulant and anti-thrombotic during pregnancy, and minimises the risk of blood coagulation in the placental circulation and fetal loss. Placental annexin A5 is prerequisite for intact pregnancy.[br][br]Nothing to Disclose: MK, HU, YN, DR, TY, SK, YH, BB, EP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 810 173 1072 SUN-15 PO33-01 Sunday 867 2012


866 ENDO12L_SUN-16 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Endoplasmic Reticulum Stress-Unfolded Protein Response Regulates Preterm Birth Chandrashekara Nagaraj Kyathanahalli, Jeyasuria Pancharatnam, Jennifer C Condon University of Pittsburgh, Pittsburgh, PA; Magee Women[apos]s Research Institute, Pittsburgh, PA Parturition is a stimulus-signal synchronized event regulated largely by hormones and their receptors. Increased expression of oxytocin receptors and gap junction proteins with a parallel decrease in progesterone (P4) levels and a withdrawal of progesterone receptor action, primes the quiescent pregnant uterus for contraction. Thus, sustained uterine quiescence is essential to prevent preterm birth. Recently we demonstrated circulating P4 levels maintain uterine quiescence and controls labor onset by modulation of uterine caspase3 (CASP3)(1,2). We propose that the pregnant uterus upholds an integrated endoplasmic reticulum stress response (ERSR) to activate CASP3. At term an associated unfolded protein response (UPR) limits CASP3 function. The balance between ERSR and UPR maintains uterine quiescence till term. In this study we aim to manipulate these responses and alter uterine CASP3 tocolytic potential. Mice at gestational day (GD) 11 and 15 were administered a single intra-peritoneal dose of tunicamycin (TM, 0,0.04, 0.2,1 mg/kg bw) a known activator of ERSR. The timing of labor, expression of uterine ERSR- UPR and contractile associated proteins were examined at 8 and 24h post TM administration. Force transduction analysis was performed on mouse uterine strips exposed to TM in vitro. Serum was collected for P4 measurements. Induction of the ERSR by TM (0.2mg/kg bw) resulted in the onset of pre-term birth at both GD11 and 15. GD11 mice were more susceptible to TM-induced stress with the onset of pre-term birth occurring as early as 24 h, while at GD15 pre-term birth occurred only after 48h. The uterine ERSR-UPR expression profile was studied following TM administration (0, 0.04, 0.2, 1 mg/kg bw) in GD11 and GD15 mice at 8 and 24h. We confirmed that TM induced ERSR in the pregnant uterus within 8h. Surprisingly these events resulted in a massive compensatory UPR culminating in decreased CASP3 activation after 24h. The decline in ERSR and the increased UPR was associated with elevated Cx43 expression and the onset of preterm labor at GD17, 48h post TM administration. The potential of TM to induce uterine myometrial contraction was further confirmed by force transduction analysis where TM (5[mu]g/mL) increased the contractility of uterine strips from GD15 mice. Taken together, this study provides evidence that modification of the uterine ERSR and its associated UPR play a critical role in regulating uterine quiescence and the timing of labor.[br][br](1)Jeyasuria et al., Biol Reprod 2009;80:928-934. (2)Jeyasuria et al., Biol Reprod 2011;85:417-424.[br][br]Sources of Research Support: NIH 5R01HD065011-02.[br][br]Nothing to Disclose: CNK, JP, JCC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1026 173 1073 SUN-16 PO33-01 Sunday 868 2012


867 ENDO12L_SUN-17 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Progesterone Regulates the Endoplasmic Reticulum Stress Response in the Pregnant Uterus Arvind Suresh, Jeyasuria Pancharatnam, Jennifer C Condon University of Pittsburgh, Pittsburgh, PA; University of Pittsburgh, Pittsburgh, PA; Magee Womens Research Institute, Pittsburgh, PA We have previously reported that progesterone (P4) via the progesterone receptor (PR) maintains uterine quiescence during pregnancy by directly regulating uterine caspase-3. Elevated active uterine caspase-3 levels found during pregnancy functions in a reversible and non-apoptotic manner to maintain quiescence by disrupting myometrial contractile architecture. As term approaches a dramatic reduction in caspase-3 allows the restoration of uterine contractility (1,2,3).[br]The endoplasmic reticulum plays an important role in maintaining cellular homeostasis by regulating protein folding and synthesis. Prolonged perturbation of endoplasmic reticulum function triggers an Endoplasmic Reticulum Stress Response (ERSR) leading to the activation of the caspase cascade. However, the ERSR is balanced by an adaptive unfolded protein response (UPR) that can restore ER homeostasis and limit caspase activation. Herein we report that P4 and PR action in the pregnant uterus have the capacity to regulate caspase-3 activation indirectly through modification of the uterine ERSR and UPR. We speculate that P4 via the PR upregulates the ERSR and inhibits the UPR thereby activating caspase-3 to maintain uterine quiescence. The withdrawal of PR action at term abrogates the ERSR to resolve ER stress mediated caspase-3 activation allowing for the reestablishment of uterine contractility.[br]Studies were performed on uterine tissues isolated from pregnant mice, a) at E10-19. b) Following treatment with vehicle or P4 from E12-19 and c) following treatment with the PR antagonist RU486 from 0-10h at 2hr intervals.[br]Utilizing CHOP and BiP as markers of the ERSR and UPR respectively, maximal levels of CHOP activation were found to correlate with maximal uterine capsase-3 levels at mid gestation. Whereas maximal BiP levels were found to correlate with the decrease in caspase-3 activation, indicating the resolution of ER stress through the UPR as term approaches. Treatment with P4 downregulated BiP, elevated CHOP and consequently increased uterine caspase-3 activation which caused a delay in the onset of labor. Conversely, treatment with RU486 resulted in an upregulation of BiP and a rapid downregulation of caspase-3 levels and the induction of preterm birth.[br]These results suggest that progesterone through its receptor mediated action regulates uterine caspase-3 activation by modulating the balance between the ERSR and the UPR to maintain appropriate gestational timing.[br][br](1)Jeyasuria P et al., Biol Reprod 2009; 80(5):928-34. (2)Jeyasuria P et al., Biol Reprod 2011; 85 (2) :417-24. (3)Stephenson-Famy A et al., Endocrinology 2012; 26(2):320-30.[br][br]Sources of Research Support: March of Dimes 21-FY2008-555. NIH 5R01HD065011-02.[br][br]Nothing to Disclose: AS, JP, JCC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 917 173 1074 SUN-17 PO33-01 Sunday 869 2012


868 ENDO12L_SUN-18 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Role of Transforming Growth Factor [beta] Type 1 Receptor in Mouse Uterus Qinglei Li, Chunjin Li, Yang Gao Texas A[amp]M University, College Station, TX Transforming growth factor [beta] (TGF[beta]) family members play an essential role in female reproduction. It has been known that TGF[beta] ligands and receptors are expressed in the uterus. However, the functional requirement of TGF[beta] signaling in the uterus is not well defined. In a recent study, we conditionally deleted TGF[beta] type 1 receptor (TGFBR1) in the female reproductive tract using anti-M[uuml]llerian hormone receptor type 2 (Amhr2)-Cre to probe its reproductive function (1). The [italic]Tgfbr1[/italic] conditional knockout (cKO) mice are infertile and develop striking smooth muscle defects in the female reproductive tract (1). To further explore the role of TGF[beta] signaling in the uterus, uteri from [italic]Tgfbr1[/italic] cKO mice at different stages were examined by immunostaining using antibodies against calponin 1, cytokeratin 8, and vimentin to specify the smooth muscle, epithelium, and stroma, respectively. In contrast to the early defects in smooth muscle formation, endometrial abnormalities were more evident in older [italic]Tgfbr1[/italic] cKO females and were exacerbated with age. In contrast to controls, [italic]Tgfbr1[/italic] cKO uteri comprised cystic (single layers of flattened epithelium) and irregular glands. These glands generally consisted of abnormal epithelial cells, some of which lost nuclear polarity and contained pleomorphic nuclei. Occasional endometrial polyps could be found in [italic]Tgfbr1[/italic] cKO mice, with multilayered proliferations of epithelial cells containing pleomorphic nuclei. Moreover, loss of intervening stroma among glands was detected in [italic]Tgfbr1[/italic] cKO mice. The aforementioned phenotype resembles endometrial hyperplasia, which is a pathological condition characterized by excessive endometrial cell proliferation and a premalignant lesion/risk factor of endometrial carcinoma. Therefore, our results suggest TGF[beta] signaling plays an important role in normal uterine development and function. Our [italic]Tgfbr1[/italic] cKO mouse may thus represent a valuable model to exploit the pathogenesis of endometrial hyperplasia.[br][br](1) Li Q et al.,PLoS Genetics 2011; 7:e1002320.[br][br]Sources of Research Support: Texas A[amp]M new faculty startup funds.[br][br]Nothing to Disclose: QL, CL, YG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2179 173 1075 SUN-18 PO33-01 Sunday 870 2012


869 ENDO12L_SUN-19 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Knockdown of Integrin [beta]1 Reduced Leiomyomal Cell Spreading, Stress Fiber Formation and Proliferation Hsiu-Mei Chen, I-Hsuan Lin, Ya-Min Cheng, Lih-Yuh C Wing National Cheng Kung University Medical College, Tainan, Taiwan; National Cheng Kung University Medical College, Tainan, Taiwan; National Cheng Kung University Medical College, Tainan, Taiwan Leiomyoma, a uterine smooth muscle tumor characterized with excessive extracellular matrix (ECM), is the most common benign tumor of women to cause hysterectomy. Integrins are the major adhesion molecules on cell surface to interact with ECM. Integrins composed of various [alpha] and [beta] subunits interact with different ECM ligands. After binding with ECM, integrins recruit various intracellular proteins to regulate the organization of actin cytoskeleton, which is associated with cell spreading, cell proliferation and migration. Whether the interaction between integrins and ECM plays any role in tumor growth of leiomyoma remains unclear. This study examined the effect of knocking down integrin [beta]1 on adhesion, spreading and proliferation of leiomyomal cells. Leiomyomal cells were isolated from leiomyoma tissue from the patients who underwent hysterectomy. Leiomyomal cells expressed both integrin [beta]1 and [beta]3 with [beta]1 being the dominant form. To investigate the role of integrin [beta]1, we transfected cells with integrin [beta]1 siRNA or scramble control. Treatment of integrin [beta]1 siRNA for three days specifically suppressed integrin [beta]1 expression without affecting integrin [beta]3 levels in leiomyomal cells. Reducing integrin [beta]1 expression did not affect cell adhesion on fibronectin or collagen. Using phalloidin staining to indicate cell spreading area, integrin [beta]1 knockdown significantly reduced leiomyomal cell spreading area up to 1 hour. Furthermore, the organization of stress fibers and cell shape also changed. Integrin-[beta]1 siRNA treatment decreased the percentage of cells exhibiting organized stress fibers in comparison with control within 1 hour. Interestingly, integrin [beta]1 knockdown had no effect on the phosphorylation levels of focal adhesion kinase, a key player in focal adhesion signaling. In addition, the long-term effect of lacking integrin [beta]1 on cell growth was evaluated for 7 days. The results showed that cell growth rate was reduced in integrin [beta]1 siRNA-treated group compared with control cells. In summary, knockdown of integrin [beta]1 reduced cell spreading, stress fiber formation and cell proliferation. The change in spreading and stress fiber organization may affect cell cycle progression.[br][br]Sources of Research Support: NSC 97-2320-B-006-019-MY3 from National Science Council, Taiwan.[br][br]Nothing to Disclose: H-MC, I-HL, Y-MC, L-YCW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1216 173 1076 SUN-19 PO33-01 Sunday 871 2012


870 ENDO12L_SUN-20 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) 17 Beta Hydroxysteroid Dehydrogenase Type 7, a Key Enzyme in Estradiol Biosynthesis, Is Highly Expressed in Cell Specific Manner in Endometriosis Y Sangeeta Devi, Anirudh Chaudhary, Susan Ferguson, Niraj Joshi, Mark Olson, Aurora Shehu, Geula Gibori, Asgi Fazleabas Michigan State University, Grand Rapids, MI; University of Illinois at Chicago, Chicago, IL Endometriosis, defined by the presence of endometrial glands and stroma in extra-uterine sites is an estrogen-dependent disease. Locally produced estradiol and aberrant expression of enzymes involved in estradiol production can promote the development of endometriosis. The principal androgen produced in female is androstendione and its conversion to estradiol involves both aromatase and HSD17B enzymes. HSD17B-7 has recently been shown to convert the weak estrone to the potent estradiol and to be highly expressed in breast cancer where it can stimulate cell growth. Whereas high expression of aromatase was reported in extrauterine endometrial tissue, that of HSD17B-7 remains totally unknown. Using a non-human primate model, we examined HSD17B-7 expression in normal endometrium, eutopic endometrium with endometriosis and extrauterine endometrial tissue. We used a baboon endometriosis model in which endometriosis was experimentally induced by intraperitoneal inoculation with menstrual endometrium. This intraperitoneal inoculation results in the formation of endometriotic lesions with gross morphological characteristics similar to those seen in women. As expected mRNA levels of aromatase, determined by by real time quantitative PCR and semi-quantitative RT-PCR was markedly increase in the endometriotic lesions as compared to normal endometrium. Interestingly, we found expression of 17bHSD-7 to be two-fold higher in eutotopic endometrium and endometriotic lesions as compared to normal endometrium without endometriosis. Localization of HSD17B-7 protein performed with a specific antibody generated against the native form of HSD17B-7 enzyme revealed that this enzyme is strongly expressed in the grandular epithelial cells. Much lower levels were detected in stromal cells of the ectopic endometrial lesions. Since aromatse has been reported to be from stromal origin, the epithelial expression of HSD17B-7 may help increase the endometriotic production of estradiol and stimulate cell growth. Taken together our results demonstrate clearly that HSD17B-7 is expressed in endometriosis and may work co-operatively with aromatase to produce estradiol de-novo.[br][br]Sources of Research Support: College of Human Medicine, MSU-YSD; HD 40093 [ndash] AF.[br][br]Nothing to Disclose: YSD, AC, SF, NJ, MO, AS, GG, AF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2315 173 1077 SUN-20 PO33-01 Sunday 872 2012


871 ENDO12L_SUN-21 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Ovarian Endometriosis Present with High Intra Tissue Estrone and Estradiol Concentration That Are Determined by Both Intracrine and Paracrine Manner Kaisa Huhtinen, Mia Stahle, Reena Desai, Anu Salminen, Antti Perheentupa, David J Handelsman, Matti Poutanen Institute of Biomedicine, University of Turku, Turku, Finland; Turku University Hospital, Turku, Finland; University of Sydney, Sydney, Australia Aberrant estrogen synthesis and metabolism has been suggested to increase local estradiol (E2) concentration in endometriosis and, thus, to promote the growth of the lesions. However, tissue estrogen concentrations within the endometrium and different types of endometriosis lesions have not been described. In the present study we evaluated intra tissue E2 and estrone (E1) concentrations in the endometrium and different types of endometriosis lesions, and correlated them with the expression of estrogen metabolizing enzymes. The data revealed that estrogen concentrations in ovarian endometriosis were remarkably higher than those measured in the endometrium and other types of endometriosis lesions. Accordingly, the the mRNA expression of the classical steroidogenic enzymes was also prominently higher in ovarian endometriotic cysts as compared to healthy endometrium or other endometriosis lesion types. Immunohistochemical staining revealed that the steroidogenic enzymes are present both in the ovarian endometriotic lesions and in the adjacent ovarian granulosa or theca cells. The data, thus, indicate that the different type of endometriotic lesions differ in their capacity to locally produce E2, and especially in the ovarian endometriosis the E2 concentration is strongly affected by local de novo synthesis. However, the ovarian lesions are also affected by sex steroids produced by paracrine manner in the ovarian tissue.[br][br]Nothing to Disclose: KH, MS, RD, AS, AP, DJH, MP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1088 173 1078 SUN-21 PO33-01 Sunday 873 2012


872 ENDO12L_SUN-22 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Telomerase Expression in Endometrium and Endometriotic Peritoneal Lesions of Infertile Women with Endometriosis Fernanda Abani Mafra, Denise Maria Christofolini, Fabia Lima Vilarino, Gustavo Mendonca Andre, Bianca Bianco, Caio Parente Barbosa Faculdade de Medicina do ABC, Santo Andre, Brazil [bold]Background: [/bold]Endometriosis has characteristics common to neoplastic cells, such as clonal proliferation, vascularization, metastasis and tissue invasion. Neoplastic cells lose the limited number of divisions due to expression of telomerase. Activation of telomerase leads to cell immortalization and studies suggested that the expression of telomerase in the functional layer of the endometrium varies according to the cyclical secretion of ovarian hormones. Many authors have reported a high telomerase activity during the proliferative phase in normal samples, while during the luteal phase, the activity is weak or absent. Based on this information, the present study aimed to evaluate the expression of telomerase in the endometrium and peritoneal endometriotic lesions from women with endometriosis and in healthy women.[br][bold]Methods:[/bold] Eleven patients with endometriosis and 44 patients without endometriosis as controls were studied. Samples of endometrium and endometriotic peritoneal lesions (only the peritoneum) were harvested in the late luteal phase of the cycle (21 [plusmn] 2 days) via laparoscopy or laparotomy using the cannula Pipelle [reg]. RNA extraction was performed with Qiazol Lysis Reagent according to the manufacturer[apos]s instructions (Qiagen). The cDNA synthesis was performed from total RNA using Super Script III kit Two-steps, according to the manufacturer[apos]s instructions (Invitrogen). The expression of [italic]hTERT [/italic]and [italic]GAPDH[/italic] genes was measured by qRT-PCR based on [italic]TaqMan[/italic] methodology (AppliedBiosystems [reg], Foster City, CA, USA). The t Student test was used to compare the values between the groups. The level of significance was p[gt]0.05.[br][bold]Results: [/bold]The [italic]hTERT[/italic] mRNA expression in the endometrium was detected in 10 (90.9%) of 11 women with endometriosis and 25 (56.8%) of 44 control women. Mean expression of [italic]hTERT[/italic] mRNA group of women with endometriosis was significantly higher when compared to the control group (10.91 [plusmn] 2.53 vs. 0.97 [plusmn] 2.52, p=0.012). When the expression of [italic]hTERT[/italic] mRNA was compared in relation to disease stage, the group of moderate/severe endometriosis showed increased expression in relation to the group of minimal/mild endometriosis (23.08 [plusmn] 2.34 vs. 0.97 [plusmn] 1.26, p=0.012).[br][bold]Conclusions:[/bold] The results of this study demonstrated an association between the expression of telomerase ([italic]hTERT[/italic] mRNA) and the genesis and/or progression of endometriosis.[br][br]Sources of Research Support: Grants from FAPESP #2010/01431-7; [bold]Acknowledgments: [/bold]The authors wish to thank FAPESP for Master in Science scholarship granted to Fernanda Abani Mafra # 2009/11298-5.[br][br]Nothing to Disclose: FAM, DMC, FLV, GMA, BB, CPB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 169 173 1079 SUN-22 PO33-01 Sunday 874 2012


873 ENDO12L_SUN-23 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) [italic]TYK2[/italic] Polymorphism Is Associated with Protection Against Endometriosis and/or Infertility Carla Peluso, Fernanda Abani Mafra, Cecilia S Goldman, Denise Maria Christofolini, Caio Parente Barbosa, Bianca Bianco Faculdade de Medicina do ABC, Santo Andre, Brazil [bold]Background[/bold]: Abnormalities in immunologic system has been suggested to be involved in the development of endometriosis. Tyrosine kinase 2 ([italic]TYK2[/italic]) has crucial importance in the etiology of autoimmune and inflammatory diseases and, recently, a number of case[ndash]control studies investigated the association of rs34536443 and rs2304256 polymorphisms with autoimmune conflicting results. Here, we aimed to evaluate [italic]TYK2[/italic] polymorphisms (rs34536443 and rs2304256) in a group of infertile women with and without endometriosis.[br][bold]Methods: [/bold]A case-control study was performed comprising 159 infertile women with endometriosis, 92 women with idiopathic infertility and 217 fertile women without history of autoimmune diseases as controls. [italic]TYK2[/italic] polymorphisms (rs34536443/C-G/Pro-Ala at exon 23 and rs2304256/A-C/Val-Phe at intron 18) were identified by [italic]TaqMan[/italic] PCR and genotype distribution, allele frequency and haplotype analysis were assessed.[br][bold]Results: [/bold]Single-marker analysis revealed that [italic]TYK2[/italic] rs34536443 was significantly associated with protection against endometriosis (p=0.001), regardless of the stage of the disease. A similar result was found to idiopathic infertility (p=0.036). When we compared infertile groups with and without endometriosis there was no statistically significant difference related to the studied polymorphism frequency (p=0.671), suggesting that the rs34536443 polymorphism is related to protection against endometriosis and/or infertility (frequency of polymorphic allele is 2.2% of endometriosis, 3.3% of idiopathic infertile and 8.1% of control group). Considering the rs2304256 polymorphism, no difference was found between endometriosis-related infertility and controls (p=0.310), even when we studied the patients with minimal/mild and moderate/severe endometriosis separately, nor to idiopathic infertility and controls (p=0.055). All groups were in Hardy-Weinberg equilibrium to both polymorphisms studied. Haplotype analysis identified a haplotype [apos][apos]CA[apos][apos] associated with protection against endometriosis and/or infertility (p=0.003 and p=0.03, respectively; frequency of 4% in endometriosis group, 8% of idiopathic infertile group and 38% of controls).[br][bold]Conclusion: [/bold]This is the first study to report an association between [italic]TYK2[/italic] polymorphisms and endometriosis and/or infertility. These findings require replication in other populations, but suggest that [italic]TYK2[/italic] rs34536443 polymorphism is associated with protection against infertility and/or endometriosis in Brazilian women studied.[br][br]Barbosa CP, et al. Frequency of endometriotic lesions in peritoneum samples from asymptomatic fertile women and correlation with CA125 values. Sao Paulo Med J 2009;127:342-5. Barrier BF. Immunology of endometriosis. Clin Obstet Gynecol 2010;53(2):397-402. Tao JH, et al. Meta-analysis of TYK2 gene polymorphisms association with susceptibility to autoimmune and inflammatory diseases. Mol Biol Rep 2011;38(7):4663-72.[br][br]Sources of Research Support: The authors wish to thank FAPESP for granting Carla Peluso (No.2011/15045-4), Fernanda Mafra (No.2009/11298-5) and Cec[iacute]lia Goldman (No.2012/00566-1) a student scholarship. This work was supported by grants No.2011/08681-1 from FAPESP.[br][br]Nothing to Disclose: CP, FAM, CSG, DMC, CPB, BB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 941 173 1080 SUN-23 PO33-01 Sunday 875 2012


874 ENDO12L_SUN-24 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) Baseline Characteristics of Endometriosis Patients Who Enrolled in 3 Clinical Trials in the U.S. Bruce Carr, Bruce Akright, Ping Jiang, Chris O[apos]Brien, Josh Burke, Roland Jimenez, John Mershon, Elizabeth Garner University of Texas Southwestern Medical Center, Dallas, TX; Northeastern Obstetrics/Gynecology Associates, Schertz, TX; Neurocrine Biosciences, San Deigo, CA; Abbott, Abbott Park, IL [bold]Introduction: [/bold]Endometriosis is a prevalent condition that primarily affects women of reproductive age. In order to further characterize patients with endometriosis, data from three recent clinical trials were pooled to examine the patients[apos] baseline characteristics.[br][bold]Methods: [/bold]Data from 3 clinical trials conducted in the US for the GnRH antagonist, elagolix were combined. This included 544 symptomatic women aged 18-49 years, who had laparoscopically confirmed endometriosis. All patients had to have a Composite Pelvic Signs and Symptoms Score (CPSSS) of [ge]6 (based on a Biberoglu and Behrman scale), and a BMI within18 to 36 kg/m[sup]2[/sup]. Baseline characteristics were summarized and the correlation between CPSSS and quality of life scores on the Endometriosis Health Profile-5 (EHP-5) at screening were assessed using Pearson correlation coefficients.[br][bold]Results: [/bold]The mean (SD) age of the patients was 31.8 [plusmn] 6.7 years, mean BMI was 26.7 [plusmn] 5.0 kg/m[sup]2[/sup], and 81% of the patients were Caucasian. Overall, 22.4%, 26.7%, 27.2%, and 9.7% had Stage I, II, III, and IV disease respectively; 14.0% had unknown disease status. The mean (SD) baseline total CPSSS score was 9.1 [plusmn] 2.0; the scores for the individual components were 2.4 [plusmn] 0.5 for dysmenorrhea, 1.8 [plusmn] 0.9 for dyspareunia, 2.1 [plusmn] 0.6 for nonmenstrual pelvic pain, 1.8 [plusmn] 0.6 for pelvic tenderness, and 1.3 [plusmn] 0.8 for pelvic induration. The total CPSSS score and EHP-5 domains of pain, control and powerlessness, social support, and self image were correlated at screening (Pearson coefficients were 0.20, 0.29, 0.20, and 0.19, respectively, p[lt]0.001 for all). The emotional well-being domain of the EHP-5 was not correlated with the total CPSSS (0.07, p=0.115).[br][bold]Conclusions: [/bold]In this combined analysis, there was an even distribution of endometriosis stages across all symptomatic women. We observed a correlation between total CPSSS score and 4 of the 5 core domains of the EHP-5, indicating a relationship between endometriosis-related pelvic pain and quality of life. A limitation of these data is that they are derived from the population of women with endometriosis who enroll in clinical trials, and therefore may not be generalisable to all endometriosis patients.[br][br]Sources of Research Support: Financial support for the three studies was provided by Neurocrine Biosciences. Elagolix is being developed by Abbott and Neurocrine Biosciences.[br][br]Disclosures: PJ: Employee, Abbott Laboratories. CO: Employee, Neurocrine Biosciences. JB: Employee, Neurocrine Biosciences. RJ: Employee, Neurocrine Biosciences; Employee, Neurocrine Biosciences. JM: Employee, Abbott Laboratories. EG: Employee, Abbott Laboratories. Nothing to Disclose: BC, BA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1582 173 1081 SUN-24 PO33-01 Sunday 876 2012


875 ENDO12L_SUN-25 POSTER SESSION: Female Reproductive Tract: Ovary, Uterus, Placenta [amp] Pregnancy (1:30 PM-3:30 PM) The Effect of Elagolix, a Novel, Orally Active GnRH Antagonist, on Bone Mineral Density (BMD) in Women with Endometriosis Bruce Ettinger, W Paul Dmowski, Chris O[apos]Brien, Martin King, Zhenzhen Xu, Josh Burke, Roland Jimenez, Elizabeth Garner, Kristof Chwalisz University of California at San Francisco, San Francisco, CA; Institute for the Study and Treatment of Endometriosis, Oak Brook, IL; Neurocrine Biosciences, San Diego, CA; Abbott, Abbott Park, IL [bold]Introduction: [/bold]GnRH agonists, including leuprolide acetate (LA), are effective treatments for endometriosis. However, long-term use without add-back therapy is limited due to hypoestrogenic effects including decreases in bone mineral density (BMD). Elagolix is a novel, oral GnRH antagonist that dose dependently suppresses estradiol (E2). We evaluated the relationship between E2 and BMD changes during treatment with LA and elagolix.[br][bold]Methods: [/bold]In this multicenter, double-blind study, Eastern European patients with endometriosis were randomized equally to receive elagolix 150 mg q.d. (n=43), elagolix 250 mg q.d. (n=44), placebo (n=43), or monthly depot LA 3.75 mg S.C. (n=44) for 12 weeks. Lumbar spine BMD was assessed by dual energy X-ray absorptiometry (DXA) at baseline and week 12. E2 levels at baseline and at the end of weeks 4, 8, and 12 were determined by LC/MS/MS; the geometric mean of monthly on-treatment plasma E2 was used in analyses. Elagolix treatment groups were combined, as results were similar by elagolix dose.[br][bold]Results: [/bold]Baseline demographics were similar among groups. Treatment with LA resulted in significantly lower on-treatment E2 than treatment with elagolix (median 5 pg/mL vs. 32 pg/mL, respectively, p[lt]0.001) and a tendency for more BMD change over 12 weeks (mean [ndash]1.6% vs. [ndash]0.9%, p=0.09).[br]In a multivariable regression model that assessed the effects of treatment, E2, baseline BMD, age, and BMI, only on-treatment E2 levels were significantly associated with spinal BMD changes (p=0.019). After adjusting for on-treatment E2 levels, there was no significant association between treatment and spinal BMD changes.[br]Overall, on-treatment E2 levels were as follows: [lt]20 pg/mL in 86% of LA-treated and 24% of elagolix-treated patients; 20-50 pg/mL in 12% of LA-treated and 49% of elagolix-treated patients; and [gt]50 pg/mL in 2% of LA-treated and 27% of elagolix-treated patients. Combining all patients, mean spinal BMD changes differed by E2 category; [ndash]1.7%, [ndash]1.2%, and +0.2%, for patients with E2 [lt]20, 20-50, and [gt]50 pg/mL, respectively.[br][bold]Conclusions: [/bold]Although the treatment duration was limited to 12 weeks, smaller decreases in spinal BMD were still observed in elagolix-treated patients compared with LA-treated patients. Overall, BMD loss was strongly and inversely correlated with E2 level, and the difference between treatment groups appeared to be due to greater suppression of E2 levels by LA.[br][br]Sources of Research Support: Financial support for the study was provided by Neurocrine Biosciences. Elagolix is being developed by Abbott and Neurocrine Biosciences.[br][br]Disclosures: BE: Consultant, Abbott Laboratories. CO: Employee, Neurocrine Biosciences. MK: Employee, Abbott Laboratories. ZX: Employee, Abbott Laboratories. JB: Employee, Neurocrine Biosciences; Employee, Neurocrine Biosciences. RJ: Employee, Neurocrine Biosciences. EG: Employee, Abbott Laboratories. KC: Employee, Abbott Laboratories. Nothing to Disclose: WPD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1626 173 1082 SUN-25 PO33-01 Sunday 877 2012


1058 ENDO12L_SUN-209 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Treatment with Sitagliptin Phosphate in Patients with Symptomatic Reactive Hypoglycemia. A Randomized, Placebo-Controlled, Double-Blinded, Clinical Trial Paloma Almeda-Valdes, Daniel Cuevas-Ramos, Griselda Brito-Cordova, Francisco J Gomez-Perez Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Mexico City, Mexico [bold]Introduction: [/bold]Sitagliptin increases glucagon-like peptide 1 circulating levels, which improves the early secretion phase (ESP) of insulin. We hypothesized that sitaglitpin could reduce symptoms of postprandial symptomatic reactive hypoglycemia (SRH). [bold]Methods: [/bold]After a two-week induction period participants were randomized to receive sitagliptin 100 mg/day (n=13) or identical placebo (n=15) for the following two weeks. SRH was defined as the presence of symptoms associated to a 2-4 hr postprandial capillary glucose [lt]65 mg/dl that disappeared with carbohydrate ingestion. Insulin and glucose during a meal tolerance test (MTT) containing 100gr of carbohydrates were compared before and after sitagliptin treatment. The areas under the curve (AUC) were calculated. The MTT was evaluated considering the early (0-30 min) and late (60-300 min) phases. Intensity of symptoms was evaluated with a visual analog scale. [bold]Results: [/bold]We studied 28 individuals (24, 85.7% women). The mean[plusmn]SD age was 34.3[plusmn]10.6 years, with a BMI of 24.6[plusmn]5.1 kg/m2. No differences in basal glucose (81.6[plusmn]7.7 vs. 81.2[plusmn]7.2 mg/dl, p=0.90) and median (interquartile range) insulin [6.3 (5.1-8.8) vs. 6.4 (5.3-10.3) [mu]U/ml, p=0.80)] were identified between groups. We did not identified differences in ESP and late secretion phase (LSP) AUC of insulin and glucose during the baseline MTT. The glucose AUC during the ESP showed a significant reduction [10263 (9380-10177) vs. 11117 (10286-11255), p=0.01) in the sitagliptin vs. control group, respectively. This was associated with an increment of the insulin AUC during ESP [2728 (1559-3025) vs. 2106 (1211-2951), p=0.01). In the LSP, a higher glucose AUC [90030 (82725-106965) vs. 84165 (73455-105015), p=0.04) with a similar insulin AUC [32805 (23250-52188) vs. 26092 (19926-40619), p=0.34] was observed in the sitagliptin vs. control group, respectively. The intensity of symptoms was significantly lower in the sitagliptin [2 (2-4)] vs. placebo group [5 (3.2-6.0)], p[lt]0.0001) after treatment. A significant reduction in anxiety, palpitations, tremor, diaphoresis, dizziness, tingling, difficulty concentrating, and weakness was demonstrated (p[lt]0.05). [bold]Conclusions: [/bold]Sitagliptin improved the ESP of insulin with a higher level of glucose in the LSP of the MTT. This biochemical change reduced the intensity of symptoms. These findings introduce a promising and new therapeutic approach for patients with reactive hypoglycemia associated to a delayed insulin secretion during the ESP.[br][br]Nothing to Disclose: PA-V, DC-R, GB-C, FJG-P 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 108 182 1265 SUN-209 PO14-01 Sunday 1061 2012


1059 ENDO12L_SUN-210 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Increased Plasma [beta]-Hydroxybutyrate Levels ([gt] 2700 [micro]mol/L) during the Fast Test in Patients with Endogenous Hyperinsulinemic Hypoglycemia Alexandre Buffet, Delphine Vezzosi, Jean-Christophe Maiza, Solange Grunenwald, Antoine Bennet, Philippe Caron CHU Larrey, Toulouse, France Context: Plasma [beta]-hydroxybutyrate (BOHB) [gt] 2700 [micro]mol/L during the 72-h fast test is thought to be the best criterion for rejecting the diagnosis of endogenous hyperinsulinemic hypoglycaemia (EHH). Objective: Characterize EHH patients with BOHB [gt] 2700 [micro]mol/L during the 72-h fast test (Gr 1) and compared them to EHH patients with BOHB [lt] 2700 [micro]mol/L (Gr 2). Design and patients: Retrospective study in 39 patients with EHH who underwent a fast test in our department. Results: During a fast test, 9 patients (including all four patients with recurrent EHH and previous partial pancreatectomy and one with previous bilateral adrenalectomy) reached BOHB [gt] 2700 [micro]mol/L (5740 +/- 176 [micro]mol/L, 2957-7824) while plasma glucose concentrations was below 3.3 mmol/L (2.26 +/- 0.68, 1.59-3.2 mmol/L). At the end of the fast test, insulin (Gr 1 = 4.75 +/- 7, 1-22 vs Gr 2 = 13.5 +/- 12, 2-58 mIU/L, p=0.004), C-peptide (Gr 1 = 0.96 +/- 1, 0.3-3.3 vs Gr 2 = 2.97 +/- 1.95, 0.8-8.9 ng/mL, p=0.0015) and pro-insulin (Gr 1 = 23.2 +/- 30.3, 2.3-92 vs Gr 2 = 121.1 +/- 120.98, 6.6-400 pmol/L, p=0.0038) levels were significantly lower in Gr 1 than in Gr 2. Plasma glucose concentrations were not different (Gr 1 = 2.64 +/- 0.67, 1.95-3.8 vs Gr 2 = 2.12 +/- 0.39, 1.5-2.8 mmol/L, p=0.0617), but the duration of fast test was longer in Gr 1 (60 +/- 16 hours, 27-72) than in Gr 2 (15 +/- 17 hours, 0-70) (p[lt]0.0001). Conclusion: Increased BOHB [gt] 2700 [micro]mol/L during the fast test can be observed in some patients with EHH, especially those with partial pancreatectomy or possible impaired glycaemic counter-regulation.[br][br]Nothing to Disclose: AB, DV, J-CM, SG, AB, PC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 204 182 1266 SUN-210 PO14-01 Sunday 1062 2012


1060 ENDO12L_SUN-211 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Study of Insulin Resistance and Associated Factors in Type 1 Diabetes Patients Monica M Teixeira, Isabella CS Arantes, Bruna P Teixeira, Danielle M Bicalho, Janice S Reis, Maria GB Castro, Teresa CA Ferrari, Maria FHS Diniz, Rodrigo B Foscolo Universidade Federal de Minas Gerais, Belo Horizonte, Brazil; Santa Casa de Belo Horizonte, Belo Horizonte, Brazil Objective: Insulin resistance (IR) is one of the pathogenic mechanisms of Type 2 diabetes. However, it can occur in patients with Type 1 diabetes and seems to be related to increased risk of cardiovascular diseases. The gold standard method for investigating and quantifying insulin resistance is the hyperinsulinemic euglycemic clamp, an invasive and expensive test. Nevertheless, IR can be estimated using some clinical and laboratory parameters. The aim of this study is to estimate IR in Type 1 Diabetes patients, according to Williams et al. (eGDR) and Dabelea et al.(LogIS)[apos]s formulas using clinical and laboratory parameters.[br]Material and Methods: This cross-sectional study was performed from January to December of 2011 with Type 1 diabetics. Protocol and formulas were described earlier. Spearman[apos]s rank correlation coefficient was used to evaluate the association between formulas and clinical parameters. All tests were two-sided and a significance level of 5% was used. Statistical analyses were conducted using SPSS for Windows version 19.[br]Results: Mean ([plusmn] SD) age, educational years and BMI ([plusmn] SD) were 31.0 [plusmn] 12.0 years, 9.3 [plusmn] 2.5 years and 23.8 [plusmn] 4.2 kg/m2, respectively. 76 (55.0%) participants were women, and the average time of diagnosis was 15.7 [plusmn] 10.5 years. The prevalence of arterial hypertension was 40%, and BMI values were 64% normal, 26% overweight and 10% obese. Among men, 90% had abdominal circumference [lt] 94cm,and 10% between 94 and 102cm. Among women, abdominal circumference values were: 51,7 %[lt]80cm, 27,6 % [gt]80[lt] 88cm, 20,7 % [ge]88cm. 29% patients(%) had A1C [lt] 7; 55% between 7,1 and 9,9, and 16% above 10. All the patients had a normal range of triglycerides, and 28,5% of women and 17,6% of men had low HDL. Median value of eGDR and LogIS were 9.82 (Q1 6.45;Q4 10.54) and 2.114 (Q1 1.813;Q4 2.291), respectively. The correlation between eGDR and LogIS was 0.689 p[lt]0.0001. There was an inverse relationship between eGDR and BMI, age, educational level, duration of diabetes. LogIS was correlated with BMI.[br]Conclusion: The results have shown a strong correlation between William et al. and Dabelea et al.[apos]s formulas, suggesting the possibility of adapting such features to clinical practice.[br][br]Nothing to Disclose: MMT, ICSA, BPT, DMB, JSR, MGBC, TCAF, MFHSD, RBF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1899 182 1267 SUN-211 PO14-01 Sunday 1063 2012


1061 ENDO12L_SUN-212 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) A First Test Effect in Intravenous Glucose Tolerance Tests Seen in Relatives of Subjects with Type 1 Diabetes Heba Ismail, Kama White, Jeffrey Krischer, Peter Chase, David Cuthbertson, Jerry P Palmer, DPT1 Study Group Seattle Children[apos]s Hospital, Seattle, WA; University of Washington, Seattle, WA; College of Medicine, University of South Florida, Tampa, FL; University of Colorado, Aurora, CO; University of South Florida, Tampa, FL The intravenous glucose tolerance test (IVGTT) is a common test of [beta]-cell function where a glucose load is administered and peripheral blood insulin and/or C-peptide responses are monitored. [bold]Objective: [/bold]Since the first IVGTT may be more stressful and stress may alter [beta]-cell secretion or hepatic insulin extraction, we asked whether there was a first test effect for either insulin or C-peptide in the IVGTT. [bold]Research Design and Methods:[/bold] Insulin and C-peptide responses were compared from two sequential IVGTTs performed during staging for the Diabetes Prevention Trial-Type 1 (DPT-1) in 368 people at high risk for type 1 diabetes. 1+3 minutes insulin data time points were used since the first phase insulin response (and peak insulin concentration) occurs within this time frame. Area under the curve (AUC) calculations estimate the insulin or C-peptide concentrations from 0 through 10 minutes post-glucose challenge. Basal levels for any given test were taken to be the average of the pre-glucose challenge values. 1+3 minute minus basal represented the sum of the 1 minute and 3 minute values minus the baseline value. [bold]Results:[/bold] More than half of all subjects were found to have first test values lower than the second. This was true for all measures of both insulin and C-peptide but was significant only for insulin 1+3 minute minus basal, AUC, and AUC minus basal, (p[lt]0.05) but not for C-peptide measures. However, for subjects (n=99) whose 1+3 minute insulin response was less than the 10[sup]th[/sup] percentile for normal on the first test, there was a significant increase in response on the second test (p[lt]0.05), while in subjects with a first test above the 10[sup]th[/sup] percentile, over 90% confirmed on the second test. The C-peptide: insulin ratio, used to investigate the mechanism of the first test effect, did not change significantly between tests, indicating that the differences seen are more likely due to changes in [beta]-cell secretion and not changes in hepatic insulin uptake. No correlation was found between age or sex and IVGTT response. [bold]Conclusions:[/bold] In summary, we found a statistically significant first test effect during the IVGTT that is most likely attributable to variations in [beta]-cell secretion of insulin rather than hepatic uptake. These differences can cause a change in an individual[apos]s eligibility for entry into a study.[br][br]Sources of Research Support: The National Institute of Diabetes and Digestive and Kidney Diseases; the National Institute of Allergy and Infectious Diseases; the Eunice Kennedy Shriver National Institute of Child Health and Human Development; the National Center for Research Resources; the American Diabetes Association; and the Juvenile Diabetes Research Foundation.[br][br]Nothing to Disclose: HI, KW, JK, PC, DC, JPP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 348 182 1268 SUN-212 PO14-01 Sunday 1064 2012


1062 ENDO12L_SUN-213 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Total Adiponectin, but Not TNF-[alpha], IL-6 or MCP-1, Correlates with the Increasing Glucose Intolerance in Pregnant Chinese-American Women Vanessa Sy, So-Young Kim, Takako Araki, Diana Huang, Anjit Khurana, Doris Tan, Emilia Liao, George Liu, Stephen Wan, Leonid Poretsky, Donna Seto-Young Beth Israel Medical Center, New York, NY; Beth Israel Medical Center, New York, NY; Beth Israel Medical Center, New York, NY [bold]Introduction:[/bold] Gestational diabetes (GDM) complicates 3.2-5% of pregnancies. Existing literature reports elevated levels of serum insulin, C-reactive protein (CRP), Tumor-necrosis factor-[alpha] (TNF-[alpha]), interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1), and decreased levels of high molecular weight adiponectin (HMW-APN) in Caucasians with GDM. Association of decreased total adiponectin (T-APN) with GDM has not been clearly established.[br][bold]Materials [amp] Methods:[/bold] 189 Chinese American women at 24-28 weeks gestation were examined for the following markers: insulin, T-APN [amp] HMW-APN, CRP, TNF-[alpha], IL-6 and MCP-1 at the time of their 50-gm glucose challenge test. Pearson correlation coefficients for glucose (1HGCT), Hemoglobin A1c (A1C), insulin and BMI were calculated against the markers described above.[br][bold]Results:[/bold] 1HGCT significantly correlated with insulin (R=0.41, p[lt]0.001) and both T-APN and HMW-APN (R= [minus]0.28, p[lt]0.001; R= [minus]0.19, p[lt]0.017). A1C significantly correlated only with T-APN (R= [minus]0.3, p[lt]0.001) and CRP (R=0.2, p[lt]0.016). Insulin significantly correlated with both T-APN and HMW-APN (R= [minus]0.23, p[lt]0.004; R= [minus]0.23, p[lt]0.003). BMI significantly correlated with CRP (R=0.27, p[lt]0.001), and both T-APN and HMW-APN (R= [minus]0.21, p[lt]0.01, R= [minus]0.26, p[lt]0.001). No significant correlations were observed between 1HGCT, A1C, insulin or BMI with TNF-[alpha], IL-6 or MCP-1.[br][bold]Conclusion:[/bold] Both T-APN [amp] HMW-APN correlate inversely with 1HGCT, insulin, and BMI in pregnant Chinese American women. In addition, T-APN inversely correlates with A1C in these women. T-APN, precursor of the active form HMW-APN, may be important in the pathogenesis for GDM. In contrast to the Caucasian GDM studies, no significant correlations were observed between the markers of glucose intolerance (1HGCT, A1C, insulin, or BMI) and inflammatory markers (TNF-[alpha], IL-6 or MCP-1). However, a weak correlation with CRP was observed. Thus, the role of inflammatory markers in the development of glucose intolerance in pregnant Chinese women may differ from their role in Caucasians. Relevance of these findings to the development of GDM in Chinese Americans remains to be established.[br][br]Nothing to Disclose: VS, S-YK, TA, DH, AK, DT, EL, GL, SW, LP, DS-Y 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 421 182 1269 SUN-213 PO14-01 Sunday 1065 2012


1063 ENDO12L_SUN-214 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Glycosylated Hemoglobin and 2[sup]nd[/sup] Hour Glucose Level after Oral Glucose Tolerance Test (OGTT) Were Significantly Higher in Individuals in the Pre-Impaired Glucose Tolerance (Pre-IGT) State Versus Those with Normal OGTT Pilar D Torres-Salvador, Chandy Lou P Malong, Mary Jane Y Tanchee-Ngo, Jay S Fonte, Bien J Matawaran, Sjoberg A Kho, Leilani B Mercado-Asis University of Santo Tomas and Faculty of Medicine and Surgery, Manila, Philippines BACKGROUND: In the initial stage of DM type 2, the [beta]-cell compensates for the insulin resistance by increasing insulin secretion to maintain normoglycemia, which we termed pre-IGT state. Our group has demonstrated that in the pre-IGT state the 2[sup]nd[/sup] hour glucose is normal ([lt]140mg/dl) but with increased insulin levels ([gt] 30uIU/ml) after 75 gm OGTT[sup](1,2)[/sup]. Clinical scoring was also developed by us which include the following risk factors: age, male gender, family history of DM and hypertension. The clinical scoring was similar in normal and pre-IGT group[sup](2)[/sup]. However, the HbA1c profile of pre-IGT subjects has not been compared those with normal OGTT. OBJECTIVES: To compare HbA1c of patients with pre-IGT to those with normal OGTT and to correlate the HbA1c with 2[sup]nd[/sup] hr glucose and insulin levels in both groups. RESEARCH METHODS: Records of 125 patients who underwent 75 gm OGTT who were evaluated for type 2 diabetes were reviewed. Demographic profile of patients such as age and BMI were determined and tabulated as mean [plusmn]SD. Only those with normal 2[sup]nd[/sup] hr glucose and insulin level (normal OGTT) and normal 2[sup]nd[/sup] hr glucose with elevated insulin level (pre-IGT) were included. The HbA1c of patients with normal OGTT was compared with the pre-IGT group using t- test. Correlation between the 2[sup]nd[/sup] hr blood glucose level and insulin with the HbA1c was done using the pearson correlation analysis. P value [lt] 0.05 was considered significant. RESULTS: Out of 125 charts reviewed, 65 patients satisfied the inclusion criteria. There were 33 subjects with normal OGTT (51%) and 32 subjects with pre-IGT (49%). The age and BMI were not significantly different between the two groups (34[plusmn]12vs.42[plusmn]14 years, 27[plusmn]6vs.29[plusmn]6 kg/m[sup]2) [/sup]respectively. Although, both in the normal range, the 2[sup]nd[/sup] hour glucose was significantly higher in the pre-IGT group (90vs128 mg/dl, p=0.045) with higher 2[sup]nd[/sup] hr insulin level (17vs.89 uIU/ml, p=0.001). The HbA1c of the pre-IGT group was also significantly higher than those in the normal OGTT group (5.15vs.6.28%, p=0.023). In this group, a positive correlation was noted in the HbA1c and 2[sup]nd[/sup] hr blood glucose level (p= 0.031) but not with the 2[sup]nd[/sup] hr insulin level (p= 0.937). This finding was not observed in the normal OGTT group. CONCLUSION: HbA1c, 2[sup]nd[/sup] hr blood glucose and 2[sup]nd[/sup] hr insulin levels were significantly higher in the pre-IGT group compared with normal OGTT. A positive correlation was demonstrated between the HbA1c and 2[sup]nd[/sup] hr glucose levels among the pre-IGT group.[br][br](1) Matawaran B, Mercado-Asis L. Comparison of Pancreatic Insulin Response to Hyperglycemia Among Filipino Subjects of Various Glycemic Status. Philippine Journal of Internal Medicine, 47: 25-30, Jan-Feb 2009. (2) Nisce I, Matawaran B, Mercado-Asis L. Clinical Risk Scoring is not Useful in Screening Patients with Pre-IGT (Hyperinsulinemic State): The Importance of 2nd-Hour Insulin Measurement of OGTT, Poster in EndoSoc USA 2011.[br][br]Nothing to Disclose: PDT-S, CLPM, MJYT-N, JSF, BJM, SAK, LBM-A 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2002 182 1270 SUN-214 PO14-01 Sunday 1066 2012


1064 ENDO12L_SUN-215 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Clinical and Molecular Characteristics of Patients with Congenital Hyperinsulinism: Two Novel Mutations in the K[sub]ATP[/sub] Channel Genes and Paternal Uniparental Isodisomy Jin Lee, Yoo-Mi Kim, Ji-Hye Seo, Minji Kang, Joo Hyun Kim, Chang Woo Jung, Gu-Hwan Kim, Jihun Kim, Beom Hee Lee, Jin-Ho Choi, Han-Wook Yoo Asan Medical Center, Seoul, Republic of Korea; Asan Medical Center, Seoul, Republic of Korea; Asan Medical Center, Seoul, Republic of Korea [bold]Purpose:[/bold] Congenital hyperinsulinism (CHI) is a rare metabolic disorder that has been categorized into diffuse and focal CHI. The focal form is caused by a heterozygous germline mutation of paternal origin in the [italic]ABCC8[/italic] or [italic]KCNJ11[/italic] genes located in the 11p15.1 region, accompanied by the somatic loss of the maternally inherited 11p15.1 to 11p15.5 region in the lesion, where two differentially imprinting regions (DMR), H19/IGF2 and KCNQ1, exist and play an important role in cellular growth and proliferation. This study was performed to elucidate the underlying molecular mechanisms of the development of pancreatic [beta]-cell hyperplasia in CHI.[br][bold]Patients and methods:[/bold] Nine infants with CHI were included in the study. All exons of the ATP-sensitive potassium channel (K[sub]ATP[/sub] channel) genes, [italic]ABCC8[/italic] and [italic]KCNJ11[/italic], [italic]GCK.[/italic] and [italic]GLUD1,[/italic] were amplified using genomic DNA and directly sequenced, and parental origin of each mutation was determined. The results were compared with the histological findings of surgically treated patients. To reveal the molecular defects in pancreatic tumor tissues, comprehensive investigations were done by multiplex ligation probe amplification (MLPA) and methylation-specific (MS)-MPLA analyses at H19/IGF2 and KCNQ1 at 11p15.5, microsatellite marker analysis on chromosome 11, and real-time PCR analysis for [italic]ABCC8[/italic] and [italic]KCNJ11[/italic].[br][bold]Results:[/bold] Germline mutations were identified in 4/9 patients (44.4%): two in [italic]ABCC8[/italic], 1 in the [italic]KCNJ11[/italic] and 1 in the [italic]GLUD1[/italic] genes. Three patients unresponsive to medical treatment underwent near-total pancreatectomy. Histological examination revealed focal hyperplasia of pancreatic [beta]-cells in two patients, and mixed diffuse and focal lesions in one patient. MLPA analyses at DMR and real-time PCR analyses for [italic]ABCC8[/italic] and [italic]KCNJ11[/italic] demonstrated the presence of two alleles at 11p15.1-15.5 in tumor tissues. On the other hand, MS-MLPA of H19/IGFs and KCNQ1 revealed abnormal methylation patterns; hypermethylation at H19/IGF2 in one patients and hypomethylation at KCNQ1 in the other two patients. Microsatellite analysis of all three patients demonstrated paternal isodisomy for chromosome 11p15.1-5 region in tumor tissues.[br][bold]Conclusions: [/bold]The results of our study indicate that paternal allelic duplication with maternal allelic loss at 11p15 region is one of major molecular mechanisms underlying adenomatous hyperplasia of pancreatic [beta]-cells in patients with CHI, and somatic imprinting alteration at DMRs at 11p15.5 plays an important role in tumor development.[br][br]Nothing to Disclose: JL, Y-MK, J-HS, MK, JHK, CWJ, G-HK, JK, BL, J-HC, H-WY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1150 182 1271 SUN-215 PO14-01 Sunday 1067 2012


1065 ENDO12L_SUN-216 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Adipose Depot[ndash]Specific Regulation of Insulin Signaling by Glucocorticoids Laura L Gathercole, Stuart A Morgan, Iwona J Bujalska, Maryam Nasiri, Paul M Stewart, Jeremy W Tomlinson University of Birmingham, Birmingham, UK Intra-abdominal adiposity is associated with insulin resistance and increased cardiovascular morbidity and mortality. Consequently, there is a need to identify factors involved in adipose tissue distribution. Patients with glucocorticoid (GC) excess, Cushing[apos]s syndrome, develop a classic phenotype characterized by central, but not peripheral, obesity. Differences in gene expression between omental (om) and subcutaneous (sc) adipose tissue have been described, however, the molecular mechanisms that underpin the differences in response to GCs have not been established. Whilst it is accepted that GCs cause global insulin resistance we have previously shown that in differentiated sc pre-adipocytes GCs enhance insulin signalling and action. In this study we compare the regulation of insulin signalling by GCs in paired human om and sc isolated intact adipocytes. The expression and phosphorylation of key components of the insulin signalling cascade (IR, IRS1 and 2, akt 2, PI3K) were measured using real-time PCR and western blotting following cortisol treatment (500nM, 24hr) and an insulin spike (50nM, 15min).[br]Cortisol treatment increased mRNA expression of IRS2 in sc (AU, 0.22[plusmn]0.07 [Ctrl], 0.62[plusmn]0.21, p[lt]0.05) but not om (AU, 0.46[plusmn]0.09 [Ctrl], 0.64[plusmn]0.17, p=ns) adipocytes. Expression of IR, IRS1, PI3K and akt 2 did not change in adipocytes from either depot. Cortisol increased insulin stimulated IR Tyr1158 phosphorylation in sc but not om adipocytes (sc: 1.68[plusmn]0.02fold, om: 0.73[plusmn]0.14fold). Consistent with increased IR activation insulin stimulated akt Ser473 phosphorylation was increased in cortisol treated sc adipocytes (1.58[plusmn]0.2fold p[lt]0.05). Interestingly GC treatment failed to regulate PKB/akt phosphorylation in om adipocytes (0.97[plusmn]0.2fold p=ns).[br]We have demonstrated adipose tissue depot specific regulation of insulin signalling by GCs. GCs enhanced insulin signalling in sc adipocytes, whilst failing to regulate insulin action in om cells. We have defined novel mechanisms, both transcriptional and post-translational, by which GCs differentially impact upon insulin action in sc and om depots. It is plausible that differential lipid flux, dependent upon the sc depot specific interaction between GCs and insulin, may underpin the adverse metabolic phenotype associated with GC excess.[br][br]Nothing to Disclose: LLG, SAM, IJB, MN, PMS, JWT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 813 182 1272 SUN-216 PO14-01 Sunday 1068 2012


1066 ENDO12L_SUN-217 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Anti-Inflammatory Action of Insulin on Adipose Tissue Paresh Dandona, Husam Ghanim, Sandeep Dhindsa, Mehul Vora, Kelly Green, Sanaa Abuaysheh, Ajay Chaudhuri State University of New York at Buffalo, Buffalo, NY Our previous work has shown that insulin exerts a rapid and potent anti-inflammatory effect on peripheral blood mononuclear cells (MNC) and a suppressive effect on the indices of oxidative, nitrosative and inflammatory stress in plasma. We have now hypothesized that insulin may also exert an anti-inflammatory action on adipose tissue. Ten obese non diabetic patients and 8 obese type 2 diabetics (T2DM) were infused with insulin (3.5 U/h) along with of 20% dextrose intravenously for 14h or with 20% dextrose or saline only in 3 separate visits one week apart. Dextrose infusion rate was adjusted to maintain blood glucose concentrations within 10 mg/dl of the baseline and were measured every 15 min. Blood samples and biopsies of abdominal fat were obtained at 0hr and 14hr. Insulin infusion suppressed plasma concentrations of CRP (by 31[plusmn]8% and 22[plusmn]7%) and SAA (by 24[plusmn]9% and 18[plusmn]8%) in obese and T2DM, respectively. There was a significant suppression of the expression of multiple pro-inflammatory genes including JNK-1, p38MAPKinase, PKC[beta]2 and TLR-4 in MNC of obese and T2DM. In addition, the expression of JNK-1, IKK[beta], PKC-[beta]2, TLR4 and CCR-2 fell significantly by27[plusmn]8%, 30[plusmn]9%, 32[plusmn]10%, 23[plusmn]7% and 34[plusmn]12%, respectively, (p[lt]0.05, for each) in adipose tissue of obese subjects and by22[plusmn]8%, 20[plusmn]7%, 28[plusmn]11%, 21[plusmn]7% and 29[plusmn]10%, respectively, (p[lt]0.05, for each) in adipose tissue of T2DM. These changes were not observed when dextrose or saline was infused alone for the same period on control days. These data demonstrate for the first time that insulin exerts an anti-inflammatory effect on adipose tissue in humans, in vivo. These data also have important implications for insulin resistance since many of these genes interfere with insulin signal transduction and their suppression could potentially reduce insulin resistance.[br][br]Disclosures: PD: Speaker, Novo Nordisk. Nothing to Disclose: HG, SD, MV, KG, SA, AC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2119 182 1273 SUN-217 PO14-01 Sunday 1069 2012


1067 ENDO12L_SUN-218 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Insulin Is an Insulin Sensitizer Sandeep Dhindsa, Husam Ghanim, Mehul Vora, Kelly Green, Chang Ling Sia, Ajay Chaudhuri, Paresh Dandona State University of New York at Buffalo, Buffalo, NY Since inflammatory mediators like TNF[alpha], IL-1[beta] and serine kinases interfere with insulin signal transduction and the insulin resistant states of obesity and type 2 diabetes (T2DM) are characterized by chronic inflammation, and since insulin exerts a rapid and potent anti-inflammatory effect, we hypothesized that insulin may suppress these inflammatory mediators and thus potentially be an insulin sensitizer. Seven obese non-diabetic and 13 obese type 2 diabetic patients were infused with 3.5 U/h of insulin for a period of 14h; 20% dextrose was co-infused to maintain blood glucose concentrations at 100mg/dl. Sequential blood samples were obtained for the measurement of the expression of inflammatory mediators in MNC and their concentrations in plasma. Compared to the baseline stabilization period of 3h, the infusion rates of glucose increased by 56% in the obese at 5h (6.7[plusmn]3.8 vs. 4.3[plusmn]2.7 mg/kg/min, p=0.007) and were maintained at a similar level up to the end of the infusion. The infusion rates in type 2 diabetics continued to increase till 8 hours, after which they remained stable till the end of the insulin infusion (3.7[plusmn]2.4 vs. 1.4[plusmn]0.8 mg/kg/min, 164% increase, p[lt]0.001). Associated with this consistent increase in the rates of infusion was the significant suppression of the protein levels of JNK-1(by 22[plusmn]6% and 19[plusmn]7%) and p38MAPKinase (by 26[plusmn]10% and 21[plusmn]12%) and in the mRNA expression of PKC[beta]2 (by 21[plusmn]5% and 17[plusmn]5%) and TLR-4 (by 25[plusmn]8% and 21[plusmn]9%) in obese and T2DM MNC, respectively. There was also the suppression (by 20[plusmn]8% and 28[plusmn]10%) in MNC obese and T2DM, respectively, of the protein levels of PTP-1B, the phosphatase which dephosphorylates the tyrosine residues on the [beta]-subunit of the insulin receptor and thus limits insulin signal transduction. Thus, the low dose infusion of insulin suppresses the expression of several inflammatory mediators which can interfere with insulin signal transduction while concomitantly increasing the required rate of glucose infusion to maintain baseline glucose concentrations. Insulin may thus be an insulin sensitizer.[br][br]Disclosures: PD: Speaker, Novo Nordisk. Nothing to Disclose: SD, HG, MV, KG, CLS, AC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2087 182 1274 SUN-218 PO14-01 Sunday 1070 2012


1068 ENDO12L_SUN-219 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Increased FDG Uptake in Association with Reduced Extremity Fat in HIV Patients Markella V Zanni, Martin Torriani, Kathleen Fitch, Eleni Stavrou, Miriam A Bredella, Ruth Lim, Aaron M Cypess, Steven Grinspoon Massachusetts General Hospital, Boston, MA; Massachusetts General Hospital, Boston, MA; Massachusetts General Hospital, Boston, MA; Joslin Diabetes Center, Beth Israel Deaconess Hospital, Boston, MA Background/Objective: HIV lipodystrophy - characterized by peripheral lipoatrophy, with or without central fat accumulation - confers increased metabolic risk[1]. Affected atrophic subcutaneous adipose tissue, on biopsy, shows a characteristic excess of apoptotic adipocytes [2] and inflammatory cells [3] coupled with deficient and defective adipocyte mitochondria [4, 5]. These morphologic changes in are thought to promote in situ insulin resistance and portend remote secondary pathology in muscle and liver. However, the functional activity of HIV lipodystrophic tissue in relation to metabolic risk has yet to be fully explored in vivo through the use of non-invasive imaging techniques. The objective of this study is to assess the relationship between FDG uptake in various fat depots and metabolic/immune parameters among subjects with HIV lipodystrophy.[br]Design/Methods: 13 lipodystrophic men on anti-retroviral therapy (ART) underwent whole-body 18F-fluorodeoxyglucose positron emission tomography (FDG-PET)/computed tomography (CT) scans and detailed metabolic/immune phenotyping. FDG uptake is reported as standardized uptake value (SUV).[br]Results: Among all subjects, there was a relatively higher SUV in the visceral adipose tissue (VAT) compared with abdominal subcutaneous adipose tissue (SAT) (0.27 [plusmn] 0.04 vs. 0.14 [plusmn] 0.02, p=0.0004). SUV in the abdominal SAT and VAT were inversely associated with percent trunk fat (r = -0.8, p = 0.001 and r = -0.8, p = 0.002), but were not related to HOMA-IR. SUV in the extremity SAT (mean SUV of the arm and leg SAT) was positively associated with HOMA-IR (r = 0.6, p = 0.02) and fasting hyperinsulinemia (r = 0.7, p 0.01), while percent extremity fat was not. Further, SUV in the extremity SAT was significantly associated with CD4 count (r = 0.6, p = 0.05). In multivariate modeling for HOMA-IR, SUV in the extremity SAT remained significant even after controlling for BMI and TNF-[alpha] (systemic inflammation) (R2 for model = 0.71, p = 0.02; SUV in the extremity SAT [beta]-estimate 12.3, p = 0.009).[br]Conclusions: In patients with HIV lipodystrophy, extremity SAT FDG uptake is increased in association with reduced extremity fat and may contribute to insulin resistance. Noninvasive assessments of in situ inflammation using FDG-PET may be useful complements to histological and gene expression analyses of metabolic dysregulation in peripheral fat among HIV+ patients.[br][br]1. Grinspoon S, Carr A. Cardiovascular risk and body-fat abnormalities in HIV-infected adults. N Engl J Med 2005; 352:48-62. 2. Domingo P, Matias-Guiu X, Pujol RM, et al. Subcutaneous adipocyte apoptosis in HIV-1 protease inhibitor-associated lipodystrophy. AIDS 1999; 13:2261-7. 3. Hammond E, McKinnon E, Nolan D. Human immunodeficiency virus treatment-induced adipose tissue pathology and lipoatrophy: prevalence and metabolic consequences. Clin Infect Dis 2010; 51:591-9. 4. Shikuma CM, Hu N, Milne C, et al. Mitochondrial DNA decrease in subcutaneous adipose tissue of HIV-infected individuals with peripheral lipoatrophy. AIDS 2001; 15:1801-9. 5. Sievers M, Walker UA, Sevastianova K, et al. Gene expression and immunohistochemistry in adipose tissue of HIV type 1-infected patients with nucleoside analogue reverse-transcriptase inhibitor-associated lipoatrophy. J Infect Dis 2009; 200:252-62.[br][br]Sources of Research Support: This work was supported by National Institutes of Health Grants 1UL1RR025758, 2P30AI060354 to Dr. Torriani., F32 DK085969 to Dr. Zanni., K23 DK081604 to Dr. Cypess, K24 DK064545 to Dr. Grinspoon, and M01-RR-01066 and 1 UL1 RR025758-01, Harvard Clinical and Translational Science Center, from the National Center for Research Resources.[br][br]Nothing to Disclose: MVZ, MT, KF, ES, MAB, RL, AMC, SG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1790 182 1275 SUN-219 PO14-01 Sunday 1071 2012


1069 ENDO12L_SUN-220 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Pancreatic Hormone and Leptin Responses to Prolonged Exercise during Follicular and Luteal Phases of the Menstrual Cycle V Daniel Castracane, Michelle Francois, Jennifer R Worley, Sharon Rogers, Reid Norman, Robert Kraemer Texas Technical University Health Sciences Center, Odessa, TX Prolonged exercise requires increased utilization of blood glucose and adjustment of glucoregulatory hormones. The midluteal phase of the menstrual cycle is associated with elevated progesterone and estrogen concentrations, and estrogen can reduce hepatic gluconeogenesis which could affect insulin concentrations. Amylin is co-secreted with insulin and controls influx of glucose into the blood to prevent hyperglycemia. [bold]Purpose: [/bold]To determine the effect of menstrual cycle stage on glucose, leptin, and pancreatic hormone responses to prolonged (90 min) treadmill exercise. [bold]Methods:[/bold] Five healthy eumenorrheic women (24.6[plusmn]5.1y; 67.4[plusmn]1kg) were monitored for 2 months to determine menstrual cycle length. A preliminary session was completed to determine exercise workloads. Then, in a fasted condition subjects completed a control trial (no exercise) and two randomized 90-min treadmill exercise trials at 60% VO[sub]2[/sub]max during the early follicular (EFX) and midluteal phase (MLX) of their menstrual cycle. Blood samples were analyzed for insulin, c-peptide, amylin, leptin, and glucagon, at rest (-30 and 0 min), during exercise (18, 36, 54, 72, [amp] 90 min) and after 20 min of recovery. Blood glucose was analyzed before, during exercise (54 and 90 min), and post-exercise (20 min). [bold]Results[/bold]: A significant ([italic]p[/italic] [lt] 0.05) time and time x trial effect occurred for glucose with greater glucose levels after 54 min of exercise in the EFX trial than the same time point in the control trial [89.2[plusmn]4.1 vs. 96.5.48 mg/dL], but no difference between the MLX and control trial. There was a time effect (p [lt] 0.05) for c-peptide and insulin with values for EFX and MLX declining from pre-values [1.23[plusmn]0.18 and 1.36[plusmn]0.61; 4.31[plusmn]2.41 and 5.68[plusmn]4.06] to 90 min exercise [1.06[plusmn]0.23 and 1.0[plusmn]0.19 ng/mL; 2.87[plusmn]1.55 and 3.37[plusmn]2.41 mU/mL, respectively]; there was no time x trial interaction. No time effect or time x trial interaction occurred for amylin and leptin. Amylin and leptin concentrations for EFX and MLX changed from pre-values [20.12[plusmn]14.2 and 15.18[plusmn]8.65; 15.19[plusmn]4.93 and 21.12[plusmn]6.50] to 90 min exercise [15.32[plusmn]10.19 and 14.14[plusmn]7.43 pM; 18.20[plusmn]6.17 and 19.67[plusmn]6.03 ng/mL, respectively]. No time effect or time x trial interaction occurred for glucagon concentrations which were consistent among trials. [bold]Conclusions:[/bold] Data suggest that stage of the menstrual cycle does not affect insulin, c-peptide, amylin, leptin, and glucagon responses to prolonged submaximal exercise, but could affect blood glucose responses.[br][br]Nothing to Disclose: VDC, MF, JRW, SR, RN, RK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1977 182 1276 SUN-220 PO14-01 Sunday 1072 2012


1070 ENDO12L_SUN-221 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Relationship between Melatonin, Insulin and Leptin Irina Sarcisovna Dzherieva, Natalya Ivanovna Volkova, Nadezhda Sergeevna Panfilova Rostov State Medical University, Rostov on Don, Russian Federation Introduction. Melatonin is a hormone that ensures the adaptation of physiological and metabolic processes of man to environmental conditions. It sets the rate of metabolic processes, determines the level of insulin resistance and leptin synthesis. Therefore a violation of melatonin secretion may lead to the development of metabolic disorders.[br]Aim and materials. To study the characteristics of the melatonin, leptin and insulin secretion in patients with metabolic syndrome were studied two groups of men: an experimental (n=25, age 44[plusmn]2 years, with verified metabolic syndrome according to criteria of IDF, 2005) and the control (n=23, age 45.1). Methods. The extent of melatonin secretion determined by its metabolite (6-sulfatoximelatonin, 6-SOMT) in urine collected at 4 am in the dark. To determine insulin, leptin, and 6-SOMT used ELISA kit. The results obtained were analyzed using the program Statistica 8.0. Results. The study revealed that the total melatonin secretion in the experimental group during the night was significantly lower than in control (p[lt]0.05). In both groups was revealed increase in urinary 6-SOMT at 4.00 am. Pearson correlation analysis revealed an inverse correlation between the content of the average power 6-SOMT in the urine at 4.00 am and insulin levels (|r|=0.38), and plasma glucose (|r|=0.37), pronounced negative correlation between peak concentrations of melatonin metabolites in the urine and the leptin level (|r|=0.77). Multiple regression analysis showed the existence of a rigid linear relationships between the 6-SOMT at 4.00 am and insulin (R=0.93), and leptin plasma levels (R=0.95). The calculation of odds ratios showed that in patients with no peak melatonin secretion risk of insulin resistance is 3 times higher than in control OR=3.0 (95% CI=1.3-7.0).[br]Conclusions. Thus: in patients with metabolic syndrome revealed violations of melatonin secretion, shown a lack of adequate physiological raises 6-SOMT at night; between the levels of melatonin, leptin and insulin secretions are the feedback, and a change in the melatonin secretion with respect to the primary impaired of insulin and leptin secretions.[br][br]Nothing to Disclose: ISD, NIV, NSP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2092 182 1277 SUN-221 PO14-01 Sunday 1073 2012


1071 ENDO12L_SUN-222 POSTER SESSION: Insulin [amp] Metabolic Action in Humans (1:30 PM-3:30 PM) Pancreatic Endocrine Function and Prevalence of Diabetes Mellitus after Pancreatic Surgery Tsuguka Shiwa, Kenichiro Uemura, Yoshiaki Murakami, Takeshi Sudo, Tomokazu Awaya, Masayasu Yoneda, Shuhei Nakanishi Hiroshima University, Hiroshima, Japan; Hiroshima University, Hiroshima, Japan [bold]Context: [/bold]The prevalence of diabetes mellitus (DM) and the influence of [beta] cell function on postoperative DM (PODM) after pancreatic surgery are not well investigated.[br][bold]Objective: [/bold]The aim of this study was to compare the prevalence of DM between pancreaticoduodenectomy (PD) and distal pancreatectomy (DP), and evaluate the influence and usability for prediction of [beta] cell function on PODM.[br][bold]Design and Methods:[/bold] Fifty patients who underwent pancreatectomy were enrolled. Assessment of [beta] cell function was performed by fasting C-peptide levels (F-CPR) and stimulated C-peptide levels with glucagon stimulation test (6min-CPR). [Delta]C-peptide ([Delta]CPR) was defined as 6min-CPR minus F-CPR. Before and six months after surgery, DM was diagnosed as fasting plasma glucose for [ge]126 mg/dl ([ge]7 mmol/l) and/or HbA1C of [ge]6.5%. The efficacies of C-peptide levels for prediction of PODM were assessed by Receiver Operating Characteristic (ROC) curve analysis. Subgroup analysis was performed in the patients with PODM to clarify the factors associated with PODM.[br][bold]Results:[/bold] Thirty-three patients underwent PD, and 17 patients underwent DP. Preoperative DM were observed in 10 patients (27%) of PD and 8 patients (47%) of DP, and there was no significant difference between groups ([italic]P[/italic]=0.195). Although there was no significant difference in postoperative F-CPR, 6min-CPR and [Delta]CPR levels between groups (PD [italic]vs.[/italic] DP, [italic]P[/italic] values. F-CPR: 1.75[plusmn]0.84 ng/ml [italic]vs.[/italic] 1.25[plusmn]0.57 ng/ml, [italic]P[/italic]=0.091, 6min-CPR: 2.72[plusmn]1.39 ng/ml [italic]vs.[/italic] 3.52[plusmn]1.92 ng/ml, [italic]P[/italic]=0.141, [Delta]CPR: 1.37[plusmn]1.00 ng/ml [italic]vs.[/italic] 1.77[plusmn]1.18 ng/ml, [italic]P[/italic]=0.249), the prevalence of PODM in DP was significantly higher than that in PD (65% [italic]vs. [/italic]33%, [italic]P[/italic]=0.035). The area under the curve (AUC) of [Delta]CPR was most efficacious indicator for prediction of PODM in two groups, AUCs of preoperative [Delta]CPR in PD and DP were 0.820(0.678-0.963) and 0.803(0.527-1.079), respectively. Lastly, in the patients with PODM, although, the postoperative [Delta]CPR in DP was significantly higher than that in PD (1.37[plusmn]0.87 ng/ml [italic]vs.[/italic] 0.64[plusmn]0.55 ng/ml, [italic]P[/italic]=0.030), postoperative HOMA-IR, BMI and amount of decrease in BMI were not significantly different between groups.[br][bold]Conclusions:[/bold] Preoperative [Delta]CPR with glucagon stimulation test might be proper indicator for the occurrence of PODM. There was significantly different prevalence of PODM by methods of pancreatectomy, thus not only [beta] cell function but also other factors except for insulin resistance and BMI might be associated with occurrence of PODM.[br][br]Nothing to Disclose: TS, KU, YM, TS, TA, MY, SN 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 72 182 1278 SUN-222 PO14-01 Sunday 1074 2012


1072 ENDO12L_SUN-225 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Growth Hormone (GH) Is Positively Associated with Hepatic Lipid Accumulation and Can Directly Activate SREBP1c Expression and Processing Manuel D Gahete, Jose Cordoba-Chacon, Chike Anadumaka, Raul M Luque, Rhonda D Kineman University of Illinois at Chicago, Chicago, IL; University of Cordoba, Cordoba, Spain GH has been reported to have both positive and negative effects on hepatic lipid accumulation. These confounding effects may be due to the fact that the models of GH excess used to date exhibit pathophysiologic changes in liver function including hepatocyte hypertrophy, proliferation and tumor formation, where liver size is almost double that of controls. In order to examine how alterations of GH within the physiological range regulate metabolic function, we have developed a mouse model with elevated endogenous GH/IGF-I levels by somatotrope-specific knockout of insulin and IGF-I signaling (HiGH mice [1]). In addition, we have generated a mouse model with reduced endogenous GH/IGF-I levels due to selective destruction of somatotropes in adults (adult-onset isolated GH deficiency or AOiGHD mice [2]). When HiGH mice are fed a high-fat or a standard chow diet, liver weight and hepatic triglyceride (TG) content is increased and hepatic TG secretion rate is decreased, compared to controls, where the livers of the AOiGHD mice show the opposite phenotype. Hepatic expression of genes that promote de novo lipogenesis (SREBP1c, ACC1, FAS, SCD1), TG synthesis (PPARgamma) and FFA uptake (CD36) are enhanced in HiGH mice and downregulated in AOiGHD mice. However, insulin levels are regulated in parallel with GH, where insulin can directly upregulate lipogenic genes in the liver. Therefore, in order to clarify the direct role that GH plays in regulating hepatic lipid metabolism, we have used primary mouse hepatocyte cultures. Our initial results demonstrate GH increases mRNA and protein levels of a key transcription factor in lipid metabolism (SREBP1c), as well as the expression of other lipogenic genes (SCD1 or CD36). Taken together, our results show that endogenous GH levels are positively associated with hepatic lipid accumulation, which may be due in part to a direct effect of GH on lipogenesis (SREBP1c and SCD1) and fatty acid uptake (CD36).[br][br]1 Gahete MD, Cordoba-Chacon J, Anadumaka CV, Lin Q, Bruning JC, Kahn CR, Luque RM, Kineman RD. 2011. Elevated GH/IGF-I, due to somatotrope-specific loss of both IGF-I and insulin receptors, alters glucose homeostasis and insulin sensitivity in a diet-dependent manner. Endocrinology 152(12):4825-37. 2 Luque RM, Lin Q, Cordoba-Chacon J, Subbaiah PV, Buch T, Waisman A, Vankelecom H, Kineman RD. 2011. Metabolic impact of adult-onset, isolated, growth hormone deficiency (AOiGHD) due to destruction of pituitary somatotropes. PLoS One 19;6(1):e15767.[br][br]Sources of Research Support: Fundacion Caja Madrid (M.D.G.); Fundacion Alfonso Martin Escudero (J.C.C); BFU2010-19300, RYC-2007-00186, CIBERObn (MICINN/FEDER), BIO-139, CTS-5051 (R.L.M); Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development Merit Award, National Institutes of Health Grants R21AG031465 and R01DK088133 (R.D.K.).[br][br]Nothing to Disclose: MDG, JC-C, CA, RML, RDK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 899 183 1279 SUN-225 PO08-01 Sunday 1075 2012


1073 ENDO12L_SUN-226 POSTER SESSION: Lipids (1:30 PM-3:30 PM) GLP-1 Inhibited Atherosclerotic Lesions in Rat Vascular Smooth Muscle Cells Induced by High Glucose and Insulin Xiaoning Zhao, Cuiping Yang, Vincent Wu, Gorge Tang, Ding Ma, Diandian He, Yan Li, Dehong Cai, Hongxiang Hui Southern Medical Univeristy, Guangzhou, China; Southern Medical Univeristy, Guangzhou, China; University of California, Los Angeles, CA Glucagon-like peptide 1 (GLP-1) is a gut incretin hormone that stimulates insulin secretion, and also known to activate anti-apoptotic signaling pathways in heart and pancreatic islets. Hyperglycemia, hyperinsulinemia and hyperlipidemia are often associated with atherosclerosis in type II diabetes. The present study was undertaken to investigate the effect of GLP-1 on cell proliferation, reactive oxygen species (ROS), lipid accumulation and inflammatory cytokines in rat vascular smooth muscle cells (VSMCs) induced by high glucose and high insulin concentration. We measured the effect of GLP-1 on VSMCs proliferation by the MTT colorimetric assay, assessed the effect of GLP-1on concentration of ROS by Flow Cytometry, and explored the effect of GLP-1 on the lipid accumulation of VSMCs in the presence of high glucose and insulin, using Oil Red O staining and quantitative measurement of intracellular cholesterol. The effect of GLP-1 on the gene expression of Sirtuin 1 and MCP-1 was examined by real-time quantifying PCR (qPCR). Furthermore, we examined the effect of GLP-1 on the production of inflammatory cytokines including IL-6, tumor necrosis factor-[alpha] and IL-8, in VSMCs using ELISA. GLP-1 decreasedROS concentrations in VSMCs mediated by high glucose, and reduced intracellular lipid accumulation in VSMCs mediated by high glucose and insulin. In addition, GLP-1 also increased Sirtuin 1 gene expression. We further confirmed that GLP-1 inhibited production of inflammatory cytokines IL-6, tumor necrosis factor-[alpha], IL-8, and monocyte chemoattractant protein-1 by VSMCs. In conclusion, our results suggest GLP-1 displays anti-atherosclerotic effects on lesions in rat vascular smooth muscle cells induced by high glucose and insulin.[br][br]Sources of Research Support: This work was supported by the Major State Basic Research Development Program of China (973 Program, No.2011CB504006), Songshan Lake Sci. [amp] Tech. Industry Park, Ph.D. Programs Foundation of Ministry of Education of China , Guangdong science and Technology Research Fund and Guangdong Department of Education Fund.[br][br]Nothing to Disclose: XZ, CY, VW, GT, DM, DH, YL, DC, HH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 422 183 1280 SUN-226 PO08-01 Sunday 1076 2012


1074 ENDO12L_SUN-227 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Thyroid Hormone-Responsive Protein (Spot14) Is Necessary for Complete De Novo Fatty Acid Synthesis Activation Michael C Rudolph, N Karl Maluf, Elizabeth A Wellberg, Andrew Lewis, Chris A Johnson, Natalie A Serkova, Robert C Murphy, Steven M Anderson University of Colorado Denver Anschutz Medical Campus, Aurora, CO; University of Colorado Denver Anschutz Medical Campus, Aurora, CO; University of Colorado Denver Anschutz Medical Campus, Aurora, CO; University of Colorado Denver Anschutz Medical Campus, Aurora, CO The mouse mammary gland is a powerful organ to study regulation of the de novo fatty acid synthesis pathway, because the transition from pregnancy to lactation represents a physiological switch when the pathway is sharply activated. The de novo fatty acid synthesis pathway contributes prominently to triglyceride fat (TAG) during milk production in lactation. However, the product of this pathway, free fatty acids, has additional biological functions including phospholipid biosynthesis, nuclear hormone signaling, and post-translational protein modification. Importantly, de novo fatty acid synthesis is also deregulated in many cancers and in metabolic disease making it relevant to a variety of human maladies. De novo fatty acid synthesis requires three sequential cytosolic enzymes: ATP citrate lyase (ACLY), Acetyl-CoA carboxylase 1 (ACC1), and Fatty acid synthase (FASN). Together, they convert carbon derived from glycolysis and amino acids into newly synthesized fatty acids. Effector proteins are also present that modify the de novo pathway to the synthetic demands of the cell, and in the mammary gland THRSP is one example. The THRSP genomic knock out mice have a lactation defect characterized by suppression of the de novo fatty acid synthesis pathway with no abnormalities gland morphology. Milk fat was reduced 40% from lactation day 10 null dams, and composition was decreased only in fatty acids known to originate from de novo synthesis. No differences in pathway gene expression or in total enzyme levels account for this phenotype. ACLY and ACC1 are regulated post-translation by phosphorylation, multimerization, and allosteric activation/inhibition by upstream metabolites or downstream products. Although some differences were observed in the phosphorylation status of these enzymes, metabolite analysis revealed levels of Acetyl-CoA and Malonyl-CoA were unchanged in the THRSP null gland. These results suggested THRSP might work downstream of ACLY and ACC1. Native gel electrophoresis showed that THRSP incorporates into multimeric complexes that contain ACLY, ACC1, and FASN. Although the biochemical function of THRSP in the mammary epithelium remains undefined, these results led to the hypothesis that THRSP directly interacts with FASN to modulate enzyme function. To test this hypothesis, we have developed a novel FASN activity assay that directly and precisely quantitates fatty acids using Gas Chromatography Mass Spectrometry.[br][br]Sources of Research Support: MCR supported by Department of Defense Breast Cancer Research predoctoral trainee award BC810596. SMA supported by National Institutes of Health PO1-HD38129. RCM supported by Colorado Clinical and Translational Sciences Institute (CCTSI) 5UL1RR025780. NAS supported by National Institutes of Health P30 CA046934-14 (Cancer Center Grant) and UL1 RR025780 (CTSA). EAW supported by Department of Defense Breast Cancer Research Program BC098051.[br][br]Nothing to Disclose: MCR, NKM, EAW, AL, CAJ, NAS, RCM, SMA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2035 183 1281 SUN-227 PO08-01 Sunday 1077 2012


1075 ENDO12L_SUN-228 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Impaired Mitochondrial Function and Cardiac Morphologic Alterations Induced by High-Fat Diet Aline Sousa Santos, Aluana Carlos Santana, Carlos Alberto Soares Costa, Anibal Sanches Moura, Celly Cristina Alves Nascimento-Saba Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brazil Introduction: Obese individuals have a higher risk of morbidity and mortality attributed to cardiovascular disease. The diet composition is one factor that predicts the cardiac phenotype in response to obesity and the type of fatty acid differentially influences the myocardial structure and function. Studies have showed that mitochondrial dysfunction is considered to play a key role in the pathogenesis of cardiac hypertrophy and failure, also the mitochondrial alterations present in heart failure indicate to defects at specific sites in electron transport chain. Thus, the aim of the study was to evaluate the effect of high fat diet, containing polyunsaturated fatty acid, on cardiac morphology and mitochondrial function. Methods: Mouse C57Bl/6, at 21days, received manipulated diet containing 7% (C: control diet) or 19% (HF: high fat diet) of soybean oil, until 135 days of age. Food intake and body mass were monitored. At the end of the experimental period, body composition was evaluated by DEXA, and the animals were sacrificed. The heart was collected, weighted and processed following the routine techniques for paraffin inclusion. The left ventricular myocardial fibers were used to analyze mitochondrial respiration by high resolution respirometry, Oroboros oxigraph. Plasma glucose (GLU), triglycerides (TG), cholesterol (CHO), HDL, LDL and VLDL were quantified. Results: Food intake did not differ between groups, but, the group HF showed higher body mass gain (13%) and trunk fat (44%). The GLU, CHO and HDL did not differ, however, TG, VLDL and LDL were increased in the group HF (45%, 49% and 24%, respectively). The heart weight was similar between the groups, though, the animals HF demonstrated ability to lower oxidation of carbohydrate (-47%) and fatty acid (-60%). The morphological analysis showed an increase of left ventricular wall thickness (29%) and the myocardial fibers seemed disorganized. Conclusion: The excess of lipids on diet reflected unfavorable alterations on body composition and lipid profile, promoting intraabdominal fat accumulation, hypertriglyceridemia and increase of VLDL and LDL. In the heart, the high fat diet determined deleterious characteristic, in view of cardiac hypertrophy, disorganization of myocardial fibers and impairment mitochondrial function of cardiomyocytes. These metabolic abnormalities may play an important role in the disease progression, leading to dyslipidemia, cardiac dysfunction and lipotoxicity in the heart.[br][br]1- Kenchaiah et al. Obesity and the risk of heart failure. N Engl J Med. 2002 Aug 1;347(5):305-13. 2- Jeckel et al. The role of dietary fatty acids in predicting myocardial structure in fat-fed rats. Lipids Health Dis. 2011 Jun 7;10:92. 3- Benderdour et al. Cardiac mitochondrial NADP+-isocitrate dehydrogenase is inactivated through 4-hydroxynonenal adduct formation: an event that precedes hypertrophy development. J Biol Chem. 2003 Nov 14;278(46):45154-9. 4- Rosca et al. Cardiac mitochondria in heart failure: decrease in respirasomes and oxidative phosphorylation. Cardiovasc Res. 2008 Oct 1;80(1):30-9.[br][br]Sources of Research Support: Faperj and CNPQ.[br][br]Nothing to Disclose: ASS, ACS, CASC, ASM, CCAN-S 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2286 183 1282 SUN-228 PO08-01 Sunday 1078 2012


1327 ENDO12L_SUN-509 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Pheochromocytomas and Paragangliomas: High Prevalence of SDH Mutations in Patients of Portuguese Institute of Oncology of Oporto Raquel G Martins, Joana Couto, Ana P Santos, Paula Soares, Joana Nunes, Jorge Lima, Isabel Torres Portuguese Institute of Oncology, Oporto, Portugal; Medical School, University of Oporto, Oporto, Portugal; IPATIMUP, Oporto, Portugal Introduction: The identification of new mutations associated with pheochromocytomas (PHEOS) and paragangliomas (PGS) has challenged the classical rule of 10% being hereditary. Currently, a mutation is seen in one third of cases; SDH mutations have been described in about 6% of apparently sporadic PHEOS and PGS, but this number may be even greater in cases of malignancy. This study aims to assess the prevalence of SDH mutations in patients with PHEOS/PGS.[br]Methods: Study of patients with PGS and PHEOS followed in the Portuguese Institute of Oncology of Oporto, tested for SDH mutations (n=30). Information on demographic and clinical aspects and family history was obtained. Mutation analysis of SDHB, SDHC and SDHD genes was done by PCR/direct sequencing in peripheral blood DNA. Data were analyzed in PASW.[br]Results: Mean age at diagnosis was 41.1[plusmn]17.2 years, 56.7% were male patients, 23(82.1%) had PGS (four in head and neck, three in the chest, one in the head, neck and chest and 16 in the abdomen), the average size of tumors was 6.6[plusmn]3.2 cm, 26.7%were malignant. The most common clinical manifestations were hypertension, excessive sweating and fatigue (56.5%), followed by pallor (47.8%) and palpitations (43.5%). The average delay of diagnosis was 50.1[plusmn]85.7 months. One patient had family history of PGS, none of PHEOS. However, 21.7% had family history of sudden death, 30.4% of acute myocardial infarction and 47.8% of stroke. Mutations were found in 66.7% of the sample: SDHB in 18 patients, SDHC in one patient and SDHD in one patient. The most common SDHB mutation was a deletion of exon one and promoter (12 patients, 66.7%).[br]Conclusions: Although most tumors were apparently sporadic, there was a high frequency of family history of possible complications of hormonal hypersecretion. The delay in diagnosis of this condition is still very high, which stresses the importance of genetic screening to identify carriers early. The prevalence of SDH mutations found in our population is higher than what is described in the literature. The selection of a higher number of malignant cases (as an oncology hospital) may explain part of this difference; the existence of a founder effect is another aspect to consider.[br][br]Nothing to Disclose: RGM, JC, APS, PS, JN, JL, IT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1295 194 1534 SUN-509 PO01-01 Sunday 1331 2012


1328 ENDO12L_SUN-510 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Genetic Study of Thai Patients with Pheochromocytoma, an Experience from Single Tertiary Center Thiti Snabboon, Natnicha Houngham, Wanna Chusrichun, Sarat Sunthornyothin Chulalongkorn University, Bangkok, Thailand Introduction: Advances in genetic study on pheochromocytoma indicate that the frequency of germline mutations associated with apparently sporadic tumor is higher than previously estimated. Several algorithms of genetic testing have been proposed; however, few data were available in Asians.[br]Objective: To analyze the genetic features of Thai patients with pheochromocytoma.[br]Methods: Forty-two consecutive patients with pheochromocytoma and/or paraganglioma were recruited in this study. Their records were analyzed regarding their age, sex, presenting symptoms and signs, hormonal tests, radiological and pathological findings and associated conditions. Genetic testing for nine susceptibility genes of pheochromocytoma/paraganglioma: RET, VHL, SDHB, SDHC, SDHD, SDHA, TMEM127, MAX and SDHAF2 was performed in all affected patients.[br]Results: Of the 42 patients, 31 had pheochromocytoma (24 unilateral and 7 bilateral lesions), 10 paraganglioma and one combined bilateral pheochromocytoma and paraganglioma. All but two of them were benign. Sequencing analysis successfully identified mutations in only patients with syndromic presentation or bilateral lesions. Syndromic group included three patients with MEN IIa and one patient with vHL syndrome. Four patients with non-syndromic bilateral lesions were found to have a mutation in vHL gene.[br]Conclusion: Due to uncommon genetic determinism in Thai patients with apparently sporadic pheochromocytoma, the genetic testing may be recommended only in those with syndromic group or bilateral adrenal lesions.[br][br]Sources of Research Support: Anandamahidol Research Fund of Division of Endocrine and Metabolism Division, Department of Medicine, Chulalongkorn University. Ratchadapisaksompoch Research Fund, Faculty of Medicine, Chulalongkorn University.[br][br]Nothing to Disclose: TS, NH, WC, SS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 327 194 1535 SUN-510 PO01-01 Sunday 1332 2012


1329 ENDO12L_SUN-511 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Pheochromocytomas and Paragangliomas: Genotype[ndash]Phenotype Correlation Raquel G Martins, Joana Couto, Ana P Santos, Paula Soares, Joana Nunes, Jorge Lima, Isabel Torres Portuguese Institute of Oncology, Oporto, Portugal; Medical School, University of Oporto, Oporto, Portugal; IPATIMUP, Oporto, Portugal Introduction: The still recent discovery of SDH mutations associated with pheochromocytomas (PHEOS) and paragangliomas (PGS) and the rarity of these tumors account for the lack of studies correlating genotype and phenotype of these neoplasms, in order to understand their behavior and adjust patient[apos]s treatment and follow-up according to the their specific risk. This study aims to evaluate possible correlations between PHEOS/PGS genotype and their presentation and behavior.[br]Methods: Study of patients with PGS and PHEOS followed in Portuguese Institute of Oncology of Oporto, tested for SDH mutations (n=30). Eighteen patients with SDHB mutation, one with SDHC, two with SDHD and a control group without mutations (n=9) were included. It was obtained information on urinary amine profile, demographics, clinical, and imaging presentation and disease progression. Mutation analysis of SDHB, SDHC and SDHD genes was done by PCR/direct sequencing in peripheral blood DNA. Data were analyzed in PASW.[br]Results: In the group with SDHB mutation (five different mutations), the average age at diagnosis was 36.2[plusmn]14.1 years, 66.7% were [male]; 88.0% presented PGS, two in head/neck (HD/NCK), three in thorax (TRX), 11 in abdomen (ABD); with an average size of 7.7[plusmn]3.5cm (no multiple tumors); 47.1% were malignant, two patients died. The patient with SDHC mutation, diagnosed at 51 years, showed only one abdominal PG, with 7cm, remaining free of disease. In the two patients ([male]) with SDHD mutation, the average age at diagnosis was 20[plusmn]1.4 years. Both had multiple PGS, the first in HD/NCK and TRX, the second in ABD, with average size 3.1[plusmn]2.1 cm. In the group without mutation, mean age was 49.2[plusmn]19.2 years, 66.6% were [female], 62.5% had PGS (two in HD/NCK, four in ABD). Solitary tumors were found in 100%, with average size 5.2[plusmn]2.7 cm; 33.3% were malignant, one patient died. In patients with SDH mutations, noradrenaline and normetanephrine were the more often increased urinary amines; dopamine was more often increased in the SDHB group. Adrenaline and metanephrine were more often increased in the control group.[br]Conclusions: Male, PGS, noradrenaline and normetanephrine hypersecretion were more frequent among patients with a mutation, multiple tumors were more common in SDHD mutation and malignant tumors (and dopamine hypersecretion) in SDHB, which is consistent with the literature. The inclusion of these results in more numerous series will enhance the knowledge in this rare condition.[br][br]Nothing to Disclose: RGM, JC, APS, PS, JN, JL, IT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1286 194 1536 SUN-511 PO01-01 Sunday 1333 2012


1330 ENDO12L_SUN-512 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Differential Expression of Selected Energy Metabolism Related Genes in Succinate Dehydrogenase B and von Hippel-Lindau Derived Pheochromocytomas and Paragangliomas Stephanie Maike Johanna Fliedner, Nina Kaludercic, Robert Wesley, Hana Hansikova, Zusana Hajkova, Jana Sladkova, Ivana Jochmanova, Hana Turkova, Nikoletta K Ledvai, Nazareno Paolocci, Jan Breza, Jiri Zeman, Hendrik Lehnert, Karel Pacak Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD; University Hospitals of Schleswig-Holstein, L[uuml]beck, Germany; Johns Hopkins Medical Institutions, Baltimore, MD; National Institutes of Health, Bethesda, MD; First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Prague, Czech Republic; School of Medicine, Comenius University, Bratislava, Slovakia (Slovak Republic) Pheochromocytomas and paragangliomas (PHEOs/PGLs) with underlying von Hippel-Lindau (VHL) or succinate dehydrogenase (SDHx) gene mutations have been reported to share a pseudohypoxic expression signature with increased glycolysis and decreased oxidative phosphorylation (OXPHOS). Nevertheless, tumor aggressiveness in both entities is distinct: metastases rarely occur in VHL-, while SDHB-derived PHEOs/PGLs often metastasize. Aim of this study was to identify characteristic differences in the pseudohypoxic features related to energy metabolism of VHL- and SDHB-derived PHEOs/PGLs.[br]Expression levels of selected energy metabolism related transcripts and proteins were evaluated for human SDHB- and VHL- derived PHEOs/PGLs. In addition, OXPHOS complex activity and reactive oxygen species levels were determined.[br]Increased glucose transporter 1 in VHL- and hexokinase 2 in VHL- and SDHB-PHEOs/PGLs compared to normal adrenal medulla supported a glycolytic phenotype, however to a lesser extent in SDHB. Increased lactate dehydrogenase A (LDHA) expression was evident for SDHB PGLs, possibly indicating a preferred conversion of pyruvate into lactate, possibly reducing the amount of starting material for the OXPHOS. In contrast, all OXPHOS subunits analyzed were more highly expressed in the majority of SDHB- compared to VHL-derived PHEOs/PGLs. OXPHOS complex activity was decreased at complex II, while complex III and citrate synthase activity were elevated in SDHB. Although we did not find evidence for elevated cytosolic reactive oxygen species, increased superoxide dismutase 2 expression may indicate aberrant electron transfer in SDHB-derived PHEOs/PGLs.[br]Our data confirm a glycolytic signature of VHL- and to a lesser extent SDHB-derived PHEOs/PGLs. Despite decreased complex II activity all OXPHOS subunits analyzed were more highly expressed in SDHB- than VHL-tissues. In addition, elevated LDHA indicates mitochondrial decoupling in SDHB-derived PHEOs/PGLs. Thus, despite a common pseudohypoxic signature, key energy metabolism related genes are differentially expressed in SDHB- and VHL-derived PHEOs/PGLs.[br]This study provides new insight into pathogenic mechanisms in aggressive human PHEOs/PGLs related to the Warburg effect, which may lead to identifying novel diagnostic and prognostic markers and potential therapeutic targets.[br][br]Sources of Research Support: This study was funded by the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD and partly by the Research Project of the Ministry of Health of the Czech Republic, Grant MZOVFN2005.[br][br]Nothing to Disclose: SMJF, NK, RW, HH, ZH, JS, IJ, HT, NKL, NP, JB, JZ, HL, KP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2352 194 1537 SUN-512 PO01-01 Sunday 1334 2012


1331 ENDO12L_SUN-513 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Metabolic Reprogramming in SDHB-Related Pheochromocytomas/Paragangliomas Nikoletta K Lendvai, Stephanie MJ Fliedner, Hana Turkova, Chun Zhang Yang, Zhengping Zhuang, Karel Pacak National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD Introduction: Pheochromocytomas (PHEOs)/paragangliomas (PGLs) are rare neural crest-derived tumors. As in many other tumors, a high rate of glycolysis is expected in PHEOs/PGLs. Glycolysis serves as the main source of energy, macromolecular biosynthesis, and antioxidant generation. PHEOs/PGLs derived from SDHB germline mutations have a high sensitivity for 18F-FDG PET, an imaging technique believed to reflect increased glycolytic activity.[br]Aim: The purpose of our study was to evaluate the glycolytic pathway in PHEOs/PGLs with SDHB mutations in comparison with those with von Hippel-Lindau (VHL) mutations and therefore identify possible therapeutic targets for their future treatment.[br]Materials and Methods: We studied four groups of PHEOs/PGLs with five samples in each, as follows: SDHB primary, SDHB metastatic, VHL tumors, and normal adrenal medulla (NAM). We evaluated the differential gene expression of glycolytic enzymes with RT-PCR. Additional experiments were performed to investigate the distribution of the tumor and stromal cells.[br]Results: The VHL-related PHEOs/PGLs showed significant upregulation of the glycolytic pathway compared to NAM. Surprisingly, there were no differences between the gene expression profiles of SDHB-related PHEOs/PGLs and NAM, with the exception of hexokinase II, phosphofructokinase L, and enolase 2, which showed a three-fold, two-fold and two-fold increase, respectively.[br]Conclusion: SDHB-related PHEOs/PGLs do not show metabolic reprogramming linked to increased glycolysis (the Warburg effect), suggesting that an alternate pathway for high tumor energy demand exists. Additionally, the high sensitivity of 18F-FDG PET for SDHB-related PHEOs/PGLs may reflect a different mechanism that is independent from glycolysis. Further studies are needed to better understand the metabolic reprogramming in SDHB-related PHEOs/PGLs.[br][br]Nothing to Disclose: NKL, SMJF, HT, CZY, ZZ, KP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2216 194 1538 SUN-513 PO01-01 Sunday 1335 2012


1332 ENDO12L_SUN-514 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Drug-Repositioning High-Throughput Screening for the Identification of New Therapeutic Options for Metastatic PHEO and PGL Alessio Giubellino, Shen Min, Jim Powers, Abdel Elkahloun, Min-Jung Lee, Jane Trepel, Tito Fojo, Arthur Tischler, Marc Ferrer, Karel Pacak Eunice Kennedy Shriver National Institute of Child Health [amp] Human Development, Bethesda, MD; National Human Genome Research Institute, Rockville, MD; Tufts Medical Center, Boston, MA; National Human Genome Research Institute, Bethesda, MD; National Cancer Institute, Bethesda, MD Drug repurposing or repositioning is an important part of drug discovery that has been more frequently used in the last few years for the implementation of new therapeutic options in oncology. We applied this paradigm in a screening of almost 4,000 drugs approved for human use, employing a model of metastatic pheochromocytoma, the most common tumor of the adrenal medulla in adults. Analysis of compound-titration series (dose-response curves) facilitated the selection of 50 molecules with potential bioactivity on pheochromocytoma. These drugs were classified based on molecular/cellular targets and signaling pathways affected, which facilitated the identification of fewer molecules of interest for pheochromocytoma, based on the prediction of relevant molecular pathways from a human microarray. We performed further investigations on selected compounds, including assays of cell proliferation, cell migration and invasion in cultured pheochromocytoma cell lines and in primary cell cultures from tissue procured from patients with pheochromocytoma and paraganglioma visiting our clinic. Our results show the potential for future evaluation of already approaved drugs in the clinical setting, using scientific mini-clinical trials.[br][br]Nothing to Disclose: AG, SM, JP, AE, M-JL, JT, TF, AT, MF, KP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2209 194 1539 SUN-514 PO01-01 Sunday 1336 2012


1333 ENDO12L_SUN-515 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Anti-Tumor Effects of Peptide Analogues Targeting Neuropeptide Hormone Receptors in Rodent Pheochromocytoma Cells Christian G Ziegler, Graeme Eisenhofer, Andrew V Schally, Linda Gebauer, Karoline Gondek, Monika Ehrhart-Bornstein, Stefan R Bornstein University of Dresden, Dresden, Germany; University of Miami Miller School of Medicine, Miami, FL Background: Malignant pheochromocytoma is a rare but potentially lethal chromaffin cell tumor with a particularly poor prognosis and currently no effective treatments. The novel targeted therapeutic approach we are pursuing here is based on our previous microarray and RT-PCR analyses, revealing altered expression of neuropeptide hormone receptors in adrenomedullary tumors and cell lines. Additionally, our work in tumor cell lines of both the adrenal cortex and medulla has shown a significant reduction of cell proliferation and survival as well as an increase in apoptosis and necrosis by employing peptide analogues that specifically bind to their expressed receptors.[br]Material and methods: Here, we could demonstrate mRNA and protein expression of somatostatin receptor subtype 2 (sst2), growth hormone-releasing hormone (GHRH) receptor and luteinizing hormone-releasing hormone (LHRH) receptor on the parental mouse pheochromocytoma cell (MPC) line as well as on the malignant mouse tumor tissue-derived (MTT) cell line. Additionally, employing various agonists and antagonists for respective peptide receptors we could demonstrate significant reductions of cell proliferation and increases in tumor cell apoptosis.[br]Results: The cytotoxic derivatives of somatostatin AN-162 and AN-238 significantly reduced cell numbers of MPC cells after 24-72h and significantly increased caspase 3/7 activation. Furthermore, we could evidence similar anti-tumor effects for GHRH antagonist MIA-602 and LHRH antagonist AN-152 on MPC cells and on MTT cells. Taking advantage of the same cell lines we are now setting up and optimize a mouse model of malignant pheochromocytoma, which will than be treated using peptide analogues, selected from in vitro studies to establish therapeutic efficacy.[br]Conclusion: This study should help to find the most effective peptide analogues with potential for future targeted treatment of neuroendocrine tumors in humans.[br][br]Nothing to Disclose: CGZ, GE, AVS, LG, KG, ME-B, SRB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 151 194 1540 SUN-515 PO01-01 Sunday 1337 2012


1334 ENDO12L_SUN-524 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) A Ligand-Binding Domain Mutation in the MDA-MB-453 Cell Line Model of Molecular Apocrine Breast Cancer Compromises Androgen Receptor Signaling Nicole L Moore, Grant Buchanan, Jonathan M Harris, Luke A Selth, Tina Bianco-Miotto, Stephen N Birrell, Lisa M Butler, Theresa E Hickey, Wayne D Tilley University of Adelaide, Adelaide, Australia; The Queen Elizabeth Hospital, Adelaide, Australia; Queensland University of Technology, Brisbane, Australia; University of Adelaide, Adelaide, Australia Recent studies suggest that molecular apocrine breast cancer is a subtype of estrogen receptor-[alpha] negative disease that is driven by androgen receptor (AR) signaling. As such, the AR is under investigation as a potential therapeutic target for these cancers. Proliferation of the MDA-MB-453 breast cancer cell line, the prototypical [italic]in vitro[/italic] model of molecular apocrine cancer, is stimulated by androgens such as 5[alpha]-dihydrotestosterone (DHT). Conversely, the synthetic progestin medroxyprogesterone acetate (MPA) inhibits proliferation of MDA-MB-453 cells via an AR-mediated mechanism. We report here that the [italic]AR[/italic] gene in MDA-MB-453 cells has a G-T transversion in exon 7, resulting in a glutamine to histidine amino acid substitution at amino acid 865 (Q865H) in the ligand binding domain. The Q865H variant AR exhibited reduced sensitivity to DHT and MPA in MDA-MB-453 cells compared to the wild type AR in transactivation assays with an AR-specific probasin-luciferase reporter but did not respond to non-androgenic ligands or antagonists. Molecular modeling and[italic] in vitro[/italic] assays revealed that the Q865H mutation disrupts ligand-induced AR intramolecular interactions, resulting in reduced stability of the MPA-AR complex compared to DHT-AR. Gene expression profiling indicated that DHT and MPA regulate distinct transcriptional programs in MDA-MB-453 cells and that genes regulated by DHT, but not MPA, are significantly associated with estrogen responsive genes and the Wnt signaling pathway. These results suggest that AR activity is compromised in MDA-MB-453 cells and that genes differentially regulated by DHT and MPA through the Q865H variant AR may contribute to the divergent proliferative effects of these hormones in MDA-MB-453 cells. Our findings provide new insight into AR function in MDA-MB-453 cells but bring into question the suitability of this cell line as a model of AR action. Moreover, these results highlight the need for development of additional cell line models to better understand AR action in molecular apocrine breast cancer.[br][br]Nothing to Disclose: NLM, GB, JMH, LAS, TB-M, SNB, LMB, TEH, WDT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1310 195 1541 SUN-524 PO26-01 Sunday 1338 2012


1335 ENDO12L_SUN-525 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) A Selective Androgen Receptor Modulator (RAD140) Induces Inflammation in Rat Right Heart Ventricle: Evidence for an Androgen-Specific, Species-Specific Mechanism Chris Miller, Maysoun Shomali, C Richard Lyttle, Gary Hattersley Radius Health, Cambridge, MA The selective androgen receptor modulator (SARM) RAD140 has been extensively characterized in preclinical assays in rats where it demonstrates androgen agonist-like effects on muscle and bone with attenuated androgen agonist effects on prostate and seminal vesicles. Unpublished preclinical toxicology studies with RAD140 indicated that it induced focal inflammation of the right ventricle in male and female Sprague-Dawley rats but not in male or female cynomolgous monkeys. This cardiac pathology has been previously reported for several steroidal androgens and appears to be caused by androgen-induced 11beta-hydroxylase inhibition in rat adrenal glands. In rats, inhibition of 11beta -hydroxylase activity leads to an increase in adrenal output of 11-deoxycorticosterone (DOC), a compound known to cause cardiac lesions in rat hearts. This mechanism is supported by our observations of elevated DOC in rats after RAD140 dosing. Additionally, we confirmed the protective effect of adrenalectomy using high dose RAD140 and testosterone propionate and found that adrenalectomized rats experienced less cardiac inflammation compared to the intact control rats. In contrast to the cardiac lesions in rats, there were no findings of cardiac inflammation in cynomolgus monkeys, even with doses of 1000 mg/kg/day and there was no consistent effect on raising plasma DOC levels above the range of detection. To our knowledge, this is the first direct comparison of androgen-induced myocarditis in rats versus primates and this is also the first description of rat cardiotoxicity reported for a nonsteroidal SARM. Due to the proposed mechanism of the rat cardiotoxicity supported by our work, we believe these findings indicate that androgen-induced myocarditis in rats is not readily extrapolated to primates, including humans.[br][br]Disclosures: CM: Employee, Radius Health. MS: Employee, Radius Health. CRL: Employee, Radius Health. GH: Employee, Radius Health. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 142 195 1542 SUN-525 PO26-01 Sunday 1339 2012


1336 ENDO12L_SUN-526 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) Anti-Inflammatory Prostaglandin 15d-PGJ[sub]2[/sub] Inactivates Androgen Receptor Signaling in Prostate Cancer Cells Sanna Kaikkonen, Ville Paakinaho, Anna-Liisa Levonen, Jorma J Palvimo University of Eastern Finland, Kuopio, Finland; University of Eastern Finland, Kuopio, Finland Inflammation that causes oxidative stress through production of reactive oxygen species (ROS) is likely to contribute to prostate cancer formation. ROS combined with defective detoxification processes leads to an unbalance in cellular redox state, causing DNA damage and misfolding of proteins. Oxidative stress also influences post-translational modifications of essential regulator proteins, such as transcription factors (TFs). In particular, accumulation of SUMO conjugates (SUMOylation) in TFs is often increased by oxidative stress. Androgen receptor (AR) functions as a hormone-activated TF that mediates the actions of androgens, playing an important role in the regulation of prostate cancer growth. Synthetic antiandrogens that directly inhibit the AR activity are therefore frequently used for the treatment of advanced prostate cancer. The AR is also a target for SUMOylation that modulates the receptor activity. Here, we show that 15-deoxy-[Delta][sup]12,14[/sup]-prostaglandin J[sub]2[/sub] (15d-PGJ[sub]2[/sub]), an endogenously produced anti-inflammatory prostaglandin, directly targets the AR and acts at low micromolar concentrations as a potent AR inhibitor, rapidly repressing AR target genes, such as [italic]FKBP51[/italic] and [italic]TMPRSS2[/italic] in prostate cancer cells. Moreover, the inhibitory effect of bicalutamide on the AR in VCaP cells is further potentiated by 15d-PGJ[sub]2[/sub], and also the residual agonistic activity of the antiandrogen on the AR was blunted by the prostaglandin. Exposure of AR positive prostate cancer cells to 15d-PGJ[sub]2[/sub] does not lead to a general inhibition of nuclear receptor activity or transcription, as under the same conditions, PPAR[gamma] is activated by the 15d-PGJ[sub]2[/sub]. Interestingly, 15d-PGJ[sub]2[/sub] also acutely induces modification of AR by SUMO-2/-3, but the inhibitory effect of 15d-PGJ[sub]2[/sub] does not rely on the SUMOylated lysine residues of the AR. ChIP assays indicate that the inhibitory effect of 15d-PGJ[sub]2[/sub] on [italic]FKBP51[/italic] and [italic]TMPRSS2[/italic] expression is paralleled with inhibition of the AR binding onto the regulatory regions of these genes. However, the DNA-binding activity of the receptor is not the sole AR function targeted by 15d-PGJ[sub]2[/sub], as the prostaglandin also blunted the androgen-dependent interaction between the AR amino and carboxy termini. In conclusion, our results identify 15d-PGJ[sub]2 [/sub]as a potent inhibitor of androgen signaling, suggesting novel possibilities in restricting the AR activity in prostate cancer cells.[br][br]Nothing to Disclose: SK, VP, A-LL, JJP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 684 195 1543 SUN-526 PO26-01 Sunday 1340 2012


1337 ENDO12L_SUN-527 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) Pharmaceutical Agents To Suppress Androgen Receptor Transcriptional Activity Sanjay N Mediwala, Erica Gonzalez, Huiying Sun, Adam Szafran, Michael A Mancini, Marco Marcelli Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Michael E DeBakey VA Medical Center, Houston, TX Background: The androgen receptor (AR) is involved in prostate cancer (PC) growth, and androgen-ablation therapy is the primary therapy if surgical resection fails or is not an option. Androgen ablation therapy, however, invariably fails, and castration resistant prostate cancer (CRPC) is a leading cause of death of American men. Truncated AR isoforms (T-ARs), which lack a ligand binding domain, but maintain functional activity, may explain the persistence of AR signaling in CRPC despite androgen ablation. These truncated AR isoforms are constitutively active, and thus allow for the growth of prostate cancer cells even in the clinical setting of androgen ablation. Based on this hypothesis, suppression of activity of the truncated androgen receptor AR-V7 could prove to be a critical aspect of future prostate cancer therapies.[br]We have generated a microscopy-based technology to study at the single cell level a number of parameters of the AR, including nuclear translocation, total AR expression, nuclear hyperspeckling, and transcriptional output. This technology (AR-HCA: androgen receptor high content analysis) was used to screen commercially available drug libraries and identified pathways important for the activation of AR-V7. We have selected pathways relevant to AR-V7 according to AR-HCA analysis to identify pan AR inhibitors (i.e. inhibitors of AR-V7, wtAR, and ART7877A) and describe three such drugs: BEZ235, an inhibitor of the PI3K signaling pathway, Obatoclax, an inhibitor of Bcl-2, and CUDC-101, a histone deacetylase/EGFR inhibitor.[br]Methods: The inhibitors were tested with RT qPCR in LAPC4 and LNCaP cell lines and with a dual luciferase assay in the PC3 cell line transfected with AR-V7 or wtAR.[br]Results: We show inhibition of wtAR and AR-V7 in the relevant cell lines with the use of the tested inhibitors.[br]Conclusions: The AR-V7 truncated AR isoform may play an important role in maintaining AR signaling in prostate cancer patients treated with androgen ablation. Agents that inhibit both wtAR and AR-V7 transcriptional activity may have clinical utility in the treatment of patients with castrate resistant prostate cancer.[br][br]Sources of Research Support: South Central VA Healthcare Network Pilot Project Grant (SM), DOD Prostate Cancer Research Program DAMD W81XWH-10-1-0390 (MM), Diana Helis Foundation (MAM [amp] MM).[br][br]Nothing to Disclose: SNM, EG, HS, AS, MAM, MM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2088 195 1544 SUN-527 PO26-01 Sunday 1341 2012


1338 ENDO12L_SUN-528 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) PI3K Inhibitors Lose Efficacy in Blocking AR Genomic Activity in Castrate-Resistant Prostate Cancer Cells Fabio Stossi, Erica Gonzalez, Huiying Sun, Michael A Mancini, Marco Marcelli Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Michael E DeBakey VA Medical Center, Houston, TX Prostate cancer (PCa) is the second most common cancer in men and requires an active androgen-androgen receptor (AR) axis for its growth. PCa is usually treated with androgen depletive treatments (ADT) consisting of LHRH agonists and AR inhibitors, which initially are very effective. However, most patients become ADT-resistant and develop castrate-resistant prostate cancer (CRPC) for which there is still no effective treatment. The pathways involved in progression to CRPC are under heavy scrutiny. Together with agonist binding, the AR exerts its full activity by being activated via kinase pathways (e.g. JAK/STAT, MAPK and PI3K/Akt). Importantly, the PI3K pathway is constitutively active in many CRPCs due to inactivation of the tumor suppressor gene PTEN. In this study we set out to analyze the effect of PI3K inhibitors on the genomic actions of AR. In LNCaP cells, we found that treatment with PI3K inhibitors (i.e. LY294002 and Wortmannin) but not inhibitors of other pathways prevented AR binding to the enhancers of some classic target genes (PSA and TMPRSS2). This was also mirrored by loss of p160 recruitment and, more importantly, mRNA output. In contrast, under conditions of PI3K inhibition no effect on AR recruitment to chromatin and target genes was observed in PC cell lines with a CRPC phenotype (i.e. C4-2 and 22RV1). Moreover, by using single cell immunofluorescence analysis we did not observe any significant change in AR protein level and intracellular localization in any of the cell lines tested. This data highlights the need for better understanding the complex interplay between intracellular signaling cascades that impinge upon AR signaling in different tumor subtypes so that the best possible treatment will be individualized for that particular type of cancer.[br][br]Sources of Research Support: DOD Prostate Cancer Research Program DAMD W81XWH-10-1-0390 (MM), Diana Helis Foundation (MAM [amp] MM).[br][br]Nothing to Disclose: FS, EG, HS, MAM, MM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2009 195 1545 SUN-528 PO26-01 Sunday 1342 2012


1339 ENDO12L_SUN-529 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) Transfected PTEN Inhibits AR-V7 Transcriptional Activity in PC3-AR-V7 Cells by Inhibiting Promoter Occupancy Huiying Sun, Sanjay Mediwala, Fabio Stossi, Michael A Mancini, Marco Marcelli Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Michael E DeBakey VA Medical Center, Houston, TX Reactivation of the androgen receptor (AR) after androgen deprivation therapy (ADT) failure is the single most dramatic event occurring to patients with prostate cancer (PC) and is the leading cause for PC transition to the castration resistant phenotype (CRPC). In the past few years, a highly expressed and constitutively active C-terminal truncated AR splice variant, AR-V7, has been discovered in CRPC patients and in PC cell lines. AR-V7 is expressed in 24% of metastatic lesions acquired from patients with CRPC, and at the protein level it is overexpressed in CRPC compared with tissue obtained from benign prostate or hormone-naive prostate cancer. The mechanism used by AR-V7 to sustain constitutive activity is extremely important to better understand the basic biology of transition to CRPC, and to conceive novel targets to treat prostate cancer. We have previously shown that pharmacologic inhibition of PI3K signaling with LY294002 and Wortmannin decreases AR-V7 transcriptional activity in a stably transfected PC3-AR-V7 cell line and in transiently transfected LNCaP cells. Because the PI3K pathway is constitutively active in many CRPC due to inactivation of the tumor suppressor gene PTEN, we hypothesized that PTEN activity could affect the transcriptional activity of AR-V7 by interfering with its interaction with AR-responsive promoters. In this study, we transfected PTEN (-) PC3-AR-V7 cells with a wt-PTEN plasmid and the AR-responsive promoter ARR2PB driving a luciferase reporter. With restoration of PTEN activity, LY294002 completely loses efficacy. Chromatin immunoprecipitation results indicate lack of AR-V7 occupancy of the ARR2PB promoter with the re-introduction of PTEN into PC3-AR-V7. These experiments identify a possible mechanism of AR-V7 constitutive activation in PTEN (-) CRPC.[br][br]Sources of Research Support: South Central VA Healthcare Network Pilot Project Grant (SM), DOD Prostate Cancer Research Program DAMD W81XWH-10-1-0390 (MM), Diana Helis Foundation (MAM [amp] MM).[br][br]Nothing to Disclose: HS, SM, FS, MAM, MM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2042 195 1546 SUN-529 PO26-01 Sunday 1343 2012


1340 ENDO12L_SUN-530 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) Differential Responsiveness of Androgen Receptor Splice Variants to Regulators of Androgen Receptor Action Ayesha A Shafi, David A Proia, Manjula Nakka, Nancy L Weigel Baylor College of Medicine, Houston, TX; Synta Pharmaceuticals Corporation, Lexington, MA Androgen ablation therapy is the most common treatment for advanced prostate cancer (PCa). Yet, a majority of patients will eventually develop castration-resistant prostate cancer (CRPC) for which there is no effective treatment. CRPC is androgen depletion resistant but androgen receptor (AR) dependent. AR is a nuclear receptor whose transcriptional activity is regulated by hormone binding to the ligand-binding domain in the C-terminus. Constitutively active AR splice variants that lack hormone-binding domains are expressed in many CRPCs. The common and unique properties of these variants versus full length AR are under investigation. Heat shock protein 90 (Hsp90) is an AR chaperone crucial for proper folding, hormone binding and transcriptional activity of AR as well as for activity of a number of kinases critical to the growth of tumors. Thus, Hsp90 is a potential therapeutic target in PCa. We have generated LNCaP cell lines with regulated expression of AR-V7 (also termed AR3), a variant containing exons 1 ,2, and 3 of AR and a small amount of unique sequence (16 aa), as well as a cell line expressing an artificially truncated AR (termed AR-NTD) lacking its hormone binding domain (aa 1-660). At optimal levels of induction, the variants stimulate expression of some AR target genes and increase cell growth. The expression of constitutively active variants indicates that new methods to inhibit AR activity that do not rely on inactivating the hormone-binding domain are needed. Using the LNCaP cell lines as well as transiently transfected HeLa cells we sought to determine if the AR variants also require Hsp90 for their activity using both a first generation Hsp90 inhibitor, geldanamycin, and a second generation Hsp90 inhibitor, ganetespib, currently being evaluated in a broad range of clinical trials including PCa. The instrinsic stability of AR-V7 is similar to the stability of full-length unliganded AR. However, both inhibitors rapidly decrease full-length AR levels, but neither reduces the levels of AR variants. Hsp90 inhibition strongly blocks full-length AR transcriptional activity but has little effect on AR variant activity measured using either an AR responsive reporter or expression of endogenous target genes. Thus, the variants are resistant to Hsp90 inhibition; although Hsp90 inhibitors will continue to inhibit growth promoting kinases and signaling through activated full length AR in CRPC, AR signaling through variants will continue.[br][br]Nothing to Disclose: AAS, DAP, MN, NLW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1362 195 1547 SUN-530 PO26-01 Sunday 1344 2012


1341 ENDO12L_SUN-531 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) Expression Level of the AR-V7 Androgen Receptor Splice Variant Alters Target Gene Specificity William Charles Krause, Zheng Xia, Ayesha Aisha Shafi, Manjula Nakka, Wei Li, Nancy Lynn Weigel Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX Androgen receptor (AR) belongs to the nuclear receptor family of transcription factors. Full-length AR (AR-FL) can be divided into 3 domains: the transactivation domain, the DNA-binding domain, and the ligand-binding domain (LBD). In castration-resistant prostate cancer (CRPC), AR is re-activated despite reduced circulating androgens, suggesting disruption of the coordination between AR domains and activity. Analyses of CRPC tumors and cell culture models led to the identification of alternatively spliced AR variants that lack the LBD. In particular, the AR-V7 variant is translated into protein, constitutively active, and preferentially expressed in CRPC. However, initial studies have yielded conflicting data on its ability to regulate AR target genes such as PSA, to function independently of AR-FL activity, and on what, if any, unique functions the variants possess. To address these issues, we generated an LNCaP cell line with inducible expression of AR-V7. In this line, AR-V7 activity is unaffected by siRNA silencing of AR-FL or by LBD antagonists. At expression levels less than or equal to AR-FL, AR-V7 induces expression of some endogenous AR targets, including PSA and TMPRSS2. Further increasing AR-V7 expression blocks induction of both genes; however, this higher AR-V7 expression is required for maximal repression of PCDH-[gamma]. To identify genes that might be differentially regulated by AR-FL and AR-V7, we examined AR targets shown by others to be coregulated by the FoxA1 transcription factor. FoxA1 interacts with AR near the LBD; thus, it may not interact with AR-V7. In the case of RASSF3 and LRIG1, FoxA1 is required for AR-dependent induction, while FoxA1 inhibits AR-dependent induction of EDN2. We find neither RASSF3 nor LRIG1 is induced by low to moderate AR-V7 expression; however, high levels of AR-V7 induce LRIG1. Conversely, EDN2 expression is not induced by AR-FL but is robustly activated by AR-V7, indicating that AR-FL and AR-V7 exhibit different patterns of gene regulation. To identify the total set of differentially regulated genes, we are using next-generation sequencing. RNA-Seq analysis of the ligand-dependent AR-FL transcriptome in LNCaP cells has identified 3606 differentially regulated genes. Additionally, preliminary analysis of these data suggests that AR-FL also regulates splicing. Similar analyses for AR-V7 are ongoing. These studies show that AR-V7 activity is a function both of its expression level and of the target gene.[br][br]Nothing to Disclose: WCK, ZX, AAS, MN, WL, NLW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1298 195 1548 SUN-531 PO26-01 Sunday 1345 2012


1342 ENDO12L_SUN-532 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) Increased Neointimal Formation after Vascular Injury in Androgen Receptor Knockout Mice Anna Sofia Wilhelmson, Johan Bourghardt-Fagman, Maria E Johansson, Karel De Gendt, Guido Verhoeven, Per Fogelstrand, Asa Tivesten University of Gothenburg, Gothenburg, Sweden; University of Gothenburg, Gothenburg, Sweden; Katholieke Universiteit Leuven, Leuven, Belgium [italic]Aim:[/italic] Testosterone is the most important sex steroid hormone in males, and evidence suggests that testosterone protects men from cardiovascular disease. We recently showed that testosterone has atheroprotective effects acting through both an androgen-receptor dependent as well as an independent pathway, and that male androgen-receptor knockout (ARKO) mice on an apolipoprotein E (ApoE)-deficient background display increased atherosclerosis. The aim of this study was to evaluate neointimal hyperplasia in ARKO mice using a model for vascular remodeling and restenosis.[br][italic]Methods:[/italic] We evaluated neointimal formation 2 weeks after ligation of the right common carotid artery in male ARKO mice and wild-type (WT) littermates on an ApoE-deficient background. The neointima was analysed for smooth muscle, macrophage, and collagen content. Also, the capacity of the endothelial layer to regenerate after a scraping injury was evaluated.[br][italic]Results:[/italic] Two weeks after carotid ligation, ARKO mice had increased total neointimal area by 104% and intimal thickness (intimal area normalized to vessel size) by 56% compared to WT mice ([italic]p[/italic][lt]0.05), while the total medial area and medial thickness did not differ between the groups. There was no difference in smooth muscle cell, macrophage, or collagen content of the neointima. Further, there was no difference in the length of endothelial regrowth in ARKO compared to WT mice.[br][italic]Conclusion:[/italic] Male ARKO mice on ApoE-deficient background display increased neointimal formation as a response to vascular injury and this increase cannot be explained by a dysfunction in the regenerative capacity of the endothelium.[br][br]Nothing to Disclose: ASW, JB-F, MEJ, KDG, GV, PF, AT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 466 195 1549 SUN-532 PO26-01 Sunday 1346 2012


1343 ENDO12L_SUN-533 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) A Functional Transgenic Androgen Receptor (AR) Rescue Model Utilizing a 240 Kb BAC Containing the Complete Mouse AR Gene Evan J Easton, Pawan Puri, John Shupe, William H Walker, Pancharatnam Jeyasuria University of Pittsburgh, Pittsburgh, PA To establish a model where a wildtype androgen receptor is replaced by mutant forms of the receptor, we have for the first time created a mouse incorporating a BAC (bacterial artificial chromosome) transgene that expresses AR in tissues that normally express AR. Testosterone targets the androgen receptor (AR) to produce the factors that are required, for male secondary sexual characteristic development, prostate function, germ cell development and survival, muscle growth and other AR related functions. The androgen receptor is linked to the X-chromosome, which makes genetic manipulation of this gene in the male difficult, as there is only a single copy of the gene. To produce this AR trangene, we used recombineering techniques to combine the relevant regions of two BACs to make a larger BAC that contained the entire AR gene plus 40 Kb each of 5[apos] and 3[apos] regulatory sequence. We then recombinereed an internal ribosome entry site (IRES) and EGFP cassette into the 3[apos] UTR of the AR transgene. The final product is a bicistronic transgene that expresses both AR and EGFP using AR regulatory sequences. Pronuclear injection of the BAC resulted in 9 founder transgenic mice of which 5 have transmitted the AR-IRES-EGFP transgene to germline. A number of the founder strains express GFP in AR expressing tissue as determined by fluorescence microscopy and western blotting. To complete the goals of our study, we have crossed mice expressing the AR-IRES-EGFP transgene with AR-floxed mice in a background of the AMH-CRE transgenic mice. Pups produced from this cross will lose AR wildtype function only in Sertoli cells. We will then determine whether the transgene is able to restore functional AR activity in the Sertoli cells of these animals. These initial studies will determine the feasibility of expressing pathway-specific AR mutants to identify the processes and factors regulated by the classical or non-classical action of testosterone in vivo. Our long-term objectives will fill a long-standing gap in the understanding of the molecular and cellular mechanisms by which testosterone acts in Sertoli cells to maintain spermatogenesis and fertility. This mouse model can also be used as the basis for understanding the genomic regulation of the AR gene.[br][br]Nothing to Disclose: EJE, PP, JS, WHW, PJ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 609 195 1550 SUN-533 PO26-01 Sunday 1347 2012


1344 ENDO12L_SUN-534 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) Molecular Changes at the Neuromuscular Junction after Nerve Injury in Response to Androgen Suggests Enhanced Neuroplasticity at the Post-Synapse in ICR Mice John Bryant, Preston E Garraghty, James T Alston, Daniel Rudmann, Gary Krishnan, Henry U Bryant, Benjamin C Yaden, Heather A Bullock, Wang X Yan, Gifondorwa David, Shetler Pamela Indiana University, Bloomington, IN; Lilly Research Laboratories, Indianapolis, IN; Lilly Research Laboratories, Indianapolis, IN The human AR is a member of the Nuclear receptor superfamily (NR3C4) located on the X chromosome and is a member of the Class I steroid receptor. The current research outlines the molecular events that occur at the post synapse, as a consequence of a unilateral focal nerve crush injury at the sciatic nerve of adult ICR mice. This focal injury results in a dramatic reduction in muscle mass and function as evidenced by the decrease in wet weight for the muscle bundle located in the gastrocnemius and at the vastus lateralis. These changes are fully reversible over time and the model represents a novel approach to study both the muscle atrophy and the repair and recovery as a consequence of focal nerve injury.[br]In this study, we utilized a TRITC coupled a-Bungarotoxin to label the acetyl choline receptors near the Neuromuscular junction (NMJ) and transcriptional changes in the nearby muscle were queried to correlate with decreased muscle mass and function subsequent to crush injury. We utilized testosterone propionate (TP) or a non-steroidal SARM to aid in recovery. The gene transcripts studied included MuSK, Rapsyn, Agrin, Neuregulin, and Dok7, mRNA all of which are components of the post synapse region of NMJ. Rapsyn expression was increased (3- 5 fold) as a result of the crush between 2 and 7 days post crush and did not seem to be modulated by SARM or TP treatment. Agrin changes (increase in 2 fold) were similar to Rapsyn and the SARM treated animals showed a sustained increase up to day 21 post crush, In contrast Dok 7 changes (increase in 2-3 fold) were evident as early as day 2 post crush and these changes were maintained only in the SARM treated animals up to day 14. The most dramatic effect was seen with MuSK mRNA, which was increased 2 fold in day 1 post crush but was further increased by both TP and SARM (4 fold) at day 1. There was a striking increase through the day 7 and 14 day period of up to 8 and 10 fold respectively. Most notably, unlike the vehicle and TP treated arms, the 10 fold increase was evident on day 21 of the SARM treated animals. We further identified the androgen regulatory region between nucleotides -1009 to -991 of the human MuSK promoter. The temporal changes and the striking fold changes in individual muscle such as the soleus, TA or gastrocnemius, in these structural components of the NMJ may point to an enhanced recovery for the androgen treated animals in this focal nerve crush animal model.[br][br]Sources of Research Support: Lilly Research Laboratories.[br][br]Nothing to Disclose: JB, PEG, JTA, DR, GK, HUB, BCY, HAB, WXY, GD, SP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2120 195 1551 SUN-534 PO26-01 Sunday 1348 2012


2232 ENDO12L_MON-700 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Cross-Sectional Studies Are Prone to Selection Bias; Example from the Danish National Study on Posttraumatic Hypopituitarism Marianne Christina Klose, Kirstine Stochholm, Marianne Andersen, Peter Laurberg, Jurgita Janukonyte, Ulla Feldt-Rasmussen Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark; Aarhus University Hospital, Aarhus, Denmark; Odense University Hospital, Odense, Denmark; Aalborg University Hospital, Aalborg, Denmark Background: Pituitary screening has been suggested in traumatic brain injury (TBI).[br]Aim: We aimed to describe the TBI population in question, and to assess the [agrave] priori selection bias.[br]Design: National cross-sectional study.[br]Patients: Patients hospitalized with a head trauma diagnosis (S06.0 [ndash] 06.9) at a Danish hospital in 2008 were identified from the Danish Board of Health diagnostic code registry. Only patients hospitalized [ge] 24h and aged 18-65 years were considered for inclusion. 2014 patients were identified and their hospital records scrutinized. Based on patient file information 1158 (58%) patients were excluded because: 1. data suggested mild TBI only, with no indication of lack of consciousness, amnesia, or cerebral imaging abnormalities (33%), 2. alcohol or drug abuse (34%), 3. death (7%) or 4. other causes (28%). The remaining 851 patients were invited to participate in the study which includes full pituitary screening; 512 (60%) agreed, 79 (9%) declined, and 265 patients (31%) did not respond to two letters.[br]Measurements: Age, gender, diagnosis, lengths and departments of hospitalization. Results: Participating patients included 337 (66%) men and 175 women, median age 44 (range 18 - 65), with a total length of hospitalization of 4 days (1-217 days). The majority had been hospitalized at a neurosurgical (40%), neurological (15%) or orthopedic (36%) department, and was registered with commotio cerebri (40%), vault or facial fractures (14%), base of skull fracture (11%), diffuse or focal brain injury (12%), traumatic subdural hemorrhage (12%), or other diagnoses of intracranial bleedings or cerebral oedema (11%). Predisposing factor for study participation included female gender (34% vs. 27%; p=0.02), older age (median 40 vs. 44 year; p=0.01), longer hospital stay (median 4,1 vs. 3.3 days; p=0.01), hospitalization at a neurosurgical ward (40% vs. 27%; p=0.003), and a more severe diagnosis, as indicated by a lower percentage of patients registered as [apos]commotio cerebri[apos] (40% vs. 50%; p=0.004).[br]Conclusion[bold]:[/bold] This study illustrates that although planned as an unselected study, systematic error may occur, here exemplified as a selection bias, caused by unequal interest to participate. This may result in an observed prevalence of posttraumatic hypopituitarism that is different from an otherwise eligible background TBI population. Such bias should be taken into consideration in all cross sectional cohort studies.[br][br]Sources of Research Support: Present and previous work within this subject was supported by unrestricted grants from: Copenhagen University, The Danish Agency for Science, Technology and Innovation, The Reseach Council of the Capital Region of Denmark, The Lundbeck Foundation, Novo Nordisk, The A.P. Moller Foundation for the Advancement of Medical Science, Arvid Nielssons Foundation, Chistenson-Cesons Foundation, Axel-Muusfeldts Foundation and Else and Mogens Wedell-Wedellsborgs Foundation.[br][br]Nothing to Disclose: MCK, KS, MA, PL, JJ, UF-R 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 738 321 2568 MON-700 PO21-02 Monday 2237 2012


2233 ENDO12L_MON-701 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Heterogeneous Post-Natal Presentation of Isolated Central Hypothyroidism (ICH) Marco Bonomi, Domenico Vladimiro Libri, Assunta Dello Iacovo, Alessandro Salvatoni, Paolo Duminico, Mario Maggi, Fabio Buzi, Giorgio Radetti, Luigi Nespoli, Annamaria De Bellis, Paolo Beck-Peccoz, Luca Persani Istituto Auxologico Italiano IRCCS and Fondazione Ca[apos] Granda Ospedale Maggiore, Milan, Italy; 2nd University of Naples, Napoli, Italy; University of Insubria, Varese, Italy; Univesrity of Florence, Firenze, Italy; [quot]A Poma[quot] Hospital, Mantova, Italy; Regional Hospital of Bolzano, Bolzano, Italy ICH may be the consequence of mutations of either TSH[beta] or TRH receptor (TRHR) genes. Recognition of genetic defect underlying ICH is often difficult due to the low number of cases reported and the lack of clear-cut diagnostic criteria. Here, we report the cases of 4 men (M) and 8 female (F), 10 sporadic and 2 familial cases, with ICH associated with low/normal TSH levels and low freeT4 levels. 1M and 1F, negative at neonatal TSH screening, showed signs of severe hypothyroidism respectively at 44 days and 4 months, while the other patients were diagnosed during childhood or adulthood (age range: 2-59 years). All, but one with a partial empty sella, had a normal pituitary MRI, and negative history of traumatic or ischemic brain injuries. Thyroid ultrasound did not uncover any sign of thyroid autoimmunity and thyroid autoantibodies were negative in all cases. Among the seven cases with blunted TSH responses to TRH stimulation but normal PRL increases a homozygous intronic mutation (IVS2+5 G[gt]A) known to alter the splicing of TSH[beta] gene was detected in one neonate with severe hypothyroidism. No alterations in TRHR gene were detected in two ICH males with combined TSH/PRL defects to TRH stimulation. Investigations of TRH gene were negative in 3 cases showing typical hypothalamic TSH responses to TRH. Since isolated gonadotropin or GH defects have been recently described in patients with anti-pituitary antibodies (APA) we decide to explore this possibility also in this ICH series by an immunofluorescence method. Interestingly, circulating APA were positive in the sera of 2 familial (mother and daughter) and 1 sporadic ICH women (at 1:32, 1:64 and 1:8 dilutions, respectively), but not in control sera. These APA positive immunofluorescence signals were localized in thyrotropes, as demonstrated by the co-localization of the anti-TSH[beta] antibody staining. These cases exhibited acquired ICH (TSH range: 0.05-0.6 mIU/L; FT4 range: 4.7-7.5 pMol/L) associated with blunted TSH and normal PRL responses to TRH stimulation test. In conclusion, idiopathic isolated CH can have a heterogeneous pathogenesis involving various genetic defects but, surprisingly, also autoimmunity. Autoimmune origin should be suspected when ICH appears to be acquired beyond infancy and the genetic origin has been excluded. This finding may open novel therapeutic perspectives for ICH as immunosuppressive treatment may be effective in restoring a normal TSH secretion.[br][br]Sources of Research Support: This work was partially supported by Ricerca Corrente Funds from Istituto Auxologico Italiano IRCCS.[br][br]Nothing to Disclose: MB, DVL, ADI, AS, PD, MM, FB, GR, LN, ADB, PB-P, LP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 897 321 2569 MON-701 PO21-02 Monday 2238 2012


2234 ENDO12L_MON-702 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Endocrine Sequelae, Sleep and Psychosocial Impairment in Adult Patients after Multimodal Treatment for Brain Tumors or Leukemia Ilonka Kreitschmann-Andermahr, Sonja Siegel, Andrea Thissen, Esther Rosenkranz, Marc Dieter Piroth, Rolf Mertens, Georg Brabant Erlangen University, Erlangen, Germany; RWTH Aachen University, Aachen, Germany; RWTH Aachen University, Aachen, Germany; RWTH Aachen University, Aachen, Germany; RWTH Aachen Unversity, Aachen, Germany; University of Luebeck, Luebeck, Germany [bold]Objective: [/bold]It is well known that multimodal brain tumor treatment may lead to endocrine, other medical and psychosocial sequelae. Yet, many countries lack tailored long-term surveillance programs for such patients so that potentially evolving health problems still may remain unrecognized. The present study was performed to investigate endocrine and psychosocial impairment in a single centre university setting in Germany in patients at least three years after treatment for brain tumours or leukaemia.[br][bold]Patients and methods: [/bold]38 (18 m, 20 f) patients were investigated at least three years after completion of brain tumor or leukemia treatment which had to include radiotherapy as one treatment modality. 26 had also been operated upon and 19 had received additional chemotherapy. 16 patients were childhood cancer survivors, 22 were diagnosed and treated as adults. In all patients basal 8:00 a.m. hormone levels (cortisol, ACTH, GH, IGF-1, fT3, fT4, TSH, LH, FSH, testosterone or estrogen) were assessed. In case of abnormal basal hormone readings or clinical suspicion of hormone abnormalities, functional endocrine assessment was additionally performed. Additionally, melatonin levels in 12 h overnight urine samples were measured in 34/38 patients. An extensive self-rating battery pertaining to quality of life, sleep disturbances and depression was also completed by all patients.[br][bold]Results: [/bold]In one patient previously undetected deficiency of three pituitary hormone axes was noted, 8 patients had abnormally low IGF-1 levels, 3 further secondary and 3 primary hypogonadism. Primary hypothyroidism, likely to be associated with brain tumor treatment (radiotherapy of the neuroaxis in medulloblastoma patients) was noted in 2 patients. Patients presented with a wide range of psychosocial impairment with women and overweight patients being more impaired than others. Higher testosterone levels were related to better psychological quality of life as indicated by the psychological sum score of the SF-36 (r=.494, p[lt].05). Patients irradiated for midline brain tumors had a significantly lower overall melatonin secretion than patients irradiated for non-midline tumors (p =.008). Urine melatonin was, however, not linked to poor sleep in self-rating questionnaires.[br][bold]Conclusion: [/bold]The results of the present study underscore the need for organized medical and psychosocial assessment and treatment of late sequelae even years after multimodal treatment for brain tumors or leukemia.[br][br]Nothing to Disclose: IK-A, SS, AT, ER, MDP, RM, GB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1249 321 2570 MON-702 PO21-02 Monday 2239 2012


2235 ENDO12L_MON-703 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Aneurysmal Subarachnoid Hemorrhage Is a Rare Cause of Acute Glucocorticoid Deficiency and Long-Term Hypopituitarism Mark J Hannon, Lucy Ann Behan, Michelle M O[apos]Brien, Karen Duggan, Mark Sherlock, Diarmuid M Smith, Amar Agha, Christopher J Thompson Beaumont Hospital/RCSI Medical School, Dublin, Ireland Subarachnoid haemorrhage (SAH) is a well reported cause of hypopituitarism but the precise incidence of hypopituitarism in this patient cohort is controversial (1-3). We aimed to prospectively determine the incidence of acute and long term hypopituitarism in SAH.[br]We prospectively recruited 100 patients (61% female, median age 53 (range 16 - 82)) with non-traumatic aneurysmal SAH. Each patient had plasma sodium, urea, osmolality, glucose, and 0900h plasma cortisol (PC) measured on days 1, 2, 3, 4, 6, 8, 10 and 12 following SAH. Results were compared with 15 patients admitted to ITU following vascular surgery. A PC [lt] 300 nmol/L in a patient in ITU was regarded clinically as inappropriately low. Survivors attended for insulin tolerance testing at [ge] 6 months following SAH. Those in whom insulin tolerance testing was contraindicated underwent glucagon stimulation testing. If patients refused either of these tests, a short synacthen test was offered.[br]14% of SAH patients had at least one PC [lt] 300nmol/L; in 4/14 (28.6%) hyponatraemia due to acute cortisol deficiency ensued, which responded to hydrocortisone treatment. In contrast, all controls had PC [gt] 500 nmol/L on day 1, and [gt] 300 nmol on days 2[ndash]12. 11% of SAH patients developed acute cranial diabetes insipidus (CDI); mortality in this group was 100%.[br]39/89 (43.8%) of SAH survivors attended for dynamic pituitary testing. The median time to testing was 15 months (range 7-30 months). 24/39 (61.5%) underwent insulin tolerance testing. 4/39 (10.3%) underwent glucagon stimulation testing and 11/39 (28.2%) underwent short synacthen testing. 2/39 (5.1%) were ACTH deficient, one of whom previously had low PC. 7/39 (17.9%) were GH deficient; one GH deficient patient previously had low PC. No patients were gonadotropin, TSH, or prolactin deficient. None had long term CDI.[br]In the largest prospective study of its kind, acute glucocorticoid deficiency occurs in 14% of SAH patients, and causes hyponatraemia in 28.6% of these. Long term hypopituitarism is uncommon following SAH and predominantly manifests as GH deficiency.[br][br]1. Aimaretti G, Ambrosio MR, Di Somma C, Gasperi M, Cannavo S, Scaroni C, et al. Residual pituitary function after brain injury-induced hypopituitarism: a prospective 12-month study. J Clin Endocrinol Metab. 2005 Nov;90(11):6085-92. 2. Klose M, Brennum J, Poulsgaard L, Kosteljanetz M, Wagner A, Feldt-Rasmussen U. Hypopituitarism is uncommon after aneurysmal subarachnoid haemorrhage. Clin Endocrinol (Oxf). 2010 Jul;73(1):95-101. 3. Parenti G, Cecchi PC, Ragghianti B, Schwarz A, Ammannati F, Mennonna P, et al. Evaluation of the Anterior Pituitary Function in the Acute Phase after Spontaneous Subarachnoid Hemorrhage. J Endocrinol Invest. 2010 Aug 31.[br][br]Nothing to Disclose: MJH, LAB, MMO, KD, MS, DMS, AA, CJT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2211 321 2571 MON-703 PO21-02 Monday 2240 2012


2236 ENDO12L_MON-704 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Do Patients with Mild Traumatic Brain Injury Have an Increased Risk for Hypopituitarism? Saadia Alvi, Sara Ahmadi, Charles R Gilkison, Randall J Urban, Brent Masel University of Texas Medical Branch, Galveston, TX; Transitional Learning Center, Galveston, TX; University of Texas Medical Branch, Galveston, TX Background:[br]An association between hypopituitarism in patients following moderate to severe Traumatic Brain Injury(TBI) has been shown. There is evidence suggesting that patients with mild TBI (mTBI) are prone to develop hypopituitarism. However, since it may not very cost effective to screen all these patients, there has been some controversy in the recommendations from the existing studies.[br]Methods:[br]Subjects were recruited from three metropolitan emergency rooms for an ongoing large consortium study of mTBI. Inclusion criteria included: closed head injury with GCS 13-15 recorded in the emergency room, loss of consciousness [lt] 30 minutes, PTA [lt] 24 hours, and absence of focal lesions on CT performed within 24 hours post-injury. Subjects underwent baseline assessments consisting of measurement of post-concussive, PTSD, and depressive symptoms, and cognition. At 6 months post-injury, pituitary function was measured by: IGF1, free and total testosterone in males, 17 beta estradiols in females, prolactin, TSH, fT4, and morning cortisols.[br]Results:[br]The study assessed 39 subjects with mild TBI available with an age of 31+/- 10 years old; ethnic composition of 33.3% Caucasians, 30.7% Latino, 30.7%African Americans, and 5.1% Asian. Blood was drawn for assessment of pituitary function on 33 of the 39 subjects at 6 months post mTBI. Of the 33 subjects, 23 were males(69.7%) and 10 were females(30.3%). All of the patients were classified as having mTBI according to a Glascow coma scale result between 14-15 and the vast majority of injuries resulted from either a motor vehicle accident, fall, or an assault. The lab results were obtained 188.5 +/- 32 days after the injury. Out of the 28 subjects who had IGF-1 levels, 53.5% (15/28) had low levels with the majority being male, 35.7% (10/28) and 17.85%(5/28) being females. In addition, when evaluating other hormone deficiencies, a low testosterone level was noted in 15% of subjects when referencing the free value and 23% when referencing the total value. All thyroid and cortisol levels were in the normal range.[br]Conclusion:[br]Assessment of pituitary function in a cohort of mostly young subjects approximately 6 months after mTBI found possible abnormalities in the reproductive and growth hormone axis. The major abnormality noted was low IGF-1 levels in both men and women. These subjects will now be assessed with a glucagon stimulations test to determine whether they have growth hormone deficiency.[br][br]Sources of Research Support: This project was part of the Mission Connect Mild TBI Translational Research Consortium, funded by the Congressionally Directed Medical Research Programs and the Department of Defense.[br][br]Nothing to Disclose: SA, SA, CRG, RJU, BM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1864 321 2572 MON-704 PO21-02 Monday 2241 2012


2237 ENDO12L_MON-705 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Comparison of Body Weight, Physical Activity and Quality of Life of Patients Following Traumatic Brain Injury or Subarachnoidal Haemorrhage with Hypopituitarism before and after Substitution of the Impaired Axis and with Normal Pituitary Function Alexandra Mueller-Oeffner, Thorsten Siegmund, Bodo Gutt, Michael Hufnagl, Petra-Maria Schumm-Draeger Academic Teaching Hospital Munich Bogenhausen, Munich, Germany; Academic Teaching Hospital Munich Schwabing, Munich, Germany; Academic Teaching Hospital Munich Bogenhausen, Munich, Germany [bold]Objective: [/bold]The prevalence of hypopituitarism following traumatic brain injury (TBI) or subarachnoidal haemorrhage (SAH) is reported between 15-55%. We analysed body weight (BW), physical activity (PA) and quality of life (QoL) of patients following TBI or SAH in relation to development and treatment of hypopituitarism.[br][bold]Method: [/bold]We diagnosed hypopituitarism in 10 patients (1 female, 9 men) of our centre (mean age 49y) who experienced a moderate or severe TBI (n=6) or SAH (n=4) in the previous 6 months by pathological basal values and dynamic testing (GHRH+ARG, ACTH, TRH or GnRH- tests) and startet a substitution therapy. The gonadotropic axis was substituted in 5, corticotropic in 3, thyreotropic in 3 and somatotropic in 2 patients for at least 4 months. Before starting and during substitution of the axis PA was assessed by pedometer and QoL was evaluated with the Nottingham Health Profile (NHP) questionnaire. We compared the results to 10 sex and aged matched patients of our centre following TBI (n=6) or SAH (n=4) without hypopituitarism.[br][bold]Results: [/bold]In comparison to patients with normal pituitary function those with hypopituitarism presented with a higher mean BMI of 4 kg/m[sup2] (p=0,043), had gained on average 5 kg BW (p=0,005) after the event and showed an average step count of less than 1750 steps (p=0,044). The QoL of patients with hypopituitarism is significantly impaired in the NHP subscale [ldquo]social isolation[rdquo] (p=0,036) and [ldquo]sleep[rdquo] (p=0,028). These subscales were significantly enhanced by substitution of the insufficient axis (p=0,044;p=0,041). The subscale [ldquo]physical activity[rdquo] and [ldquo]sleep[rdquo] was significantly increased by GH substitution in the GH deficient (p=0.0001) and the subscale [ldquo]social isolation[rdquo] by testosterone or estrogen substitution in (p=0,02) patients with secondary hypogonadism.[br][bold]Conclusion: [/bold]Patients with pituitary dysfunction gained more body weight, presented with a lower physical activity and an impairment of the quality of life since the TBI or SAH. Subsitution of the insufficient axis enhanced the quality of life.[br][br]Nothing to Disclose: AM-O, TS, BG, MH, P-MS-D 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 188 321 2573 MON-705 PO21-02 Monday 2242 2012


2238 ENDO12L_MON-706 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Pituitary Dysfunction in ANCA-Associated Vasculitides Ekta Singh, Rodrigo Cartin-Ceba, Ulrich Specks, Dana Erickson Mayo Clinic, Rochester, MN; Mayo Clinic, Rochester, MN [bold]Background: [/bold]The antineutrophil cytoplasmic antibody (ANCA)-associated vasculitides primarily involve the kidneys, lungs, and the peripheral nervous system. Pituitary gland involvement has been rarely reported. The literature on pituitary involvement in ANCA-associated vasculitides is rather sparse.[br][bold]Methods: [/bold]We reviewed the medical records of all patients with pituitary involvement in ANCA-associated vasculitides seen at Mayo Clinic, Rochester, between Jan. 1996 and Dec. 2011. We describe the clinical features, pituitary imaging findings, hormonal evaluation, and treatment response of these patients.[br][bold]Results: [/bold]Out of the 637 patients with ANCA-associated vasculitides seen between Jan.1996 and Dec.2011, pituitary involvement was noted in 8 (1.3%), without gender preponderance. The median age of diagnosis of pituitary involvement was 48 yrs (28-70). Pituitary involvement was diagnosed incidentally during imaging to evaluate headache or sinusitis in 5 patients (63%), and during evaluation of symptoms perceived to be due to pituitary dysfunction in 3 patients (37%). Other organ systems involved included ear nose and throat (100%), lungs (63%), kidney (50%), skin (25%), joints (25%), and heart (12%).[br]6 patients (75%) had diabetes insipidus (DI) during the disease course, but this was often not a presenting complaint. In terms of anterior pituitary dysfunction, 7 patients had hypogonadism, 4 had secondary hypothyroidism, 2 had hypocortisolism, 1 had elevated prolactin, and 1 had normal function. Head MRI revealed a sellar mass with peripheral enhancement or diffuse pituitary enlargement in all patients. 3 patients also had thickening of the pituitary stalk. Visual field defects were noted in 2 patients.[br]Patients were treated with standard systemic therapy. Mean follow-up duration (available for 7 patients) was 80 months (9-128). All but one patient had decrease in size of the pituitary enlargement within 1 year of treatment. One patient had progressive pituitary enlargement, and biopsy was recommended, but not pursued, and patient was lost to follow-up. DI resolved in 4 out of 6 patients. Of the 7 patients with anterior pituitary dysfunction, resolution was seen in 3, improvement in 2, and persistent dysfunction in 2.[br][bold]Conclusion[/bold]: Pituitary involvement in ANCA-associated vasculitides is rare. Majority of these patients have pituitary dysfunction, which responds variably to treatment despite improvement in pituitary imaging findings with standard systemic therapy.[br][br]Disclosures: US: Clinical Researcher, Genentech, Inc. Nothing to Disclose: ES, RC-C, DE 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1400 321 2574 MON-706 PO21-02 Monday 2243 2012


2239 ENDO12L_MON-707 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Hypothalamic Pituitary Axis Dysfunction in Patients with Meningitis and Meningoencephalitis Alok Sachan, B Vengamma, C V Harinarayan, Bindu Menon, V Suresh, M Neelima Raj Sri Venkateswara Institute of Medical Sciences, Tirupati, India; Sri Venkateswara Institute of Medical Sciences, Tirupati, India; Narayan Medical College, Nellore, India Introduction: Effect of conditions, involving central nervous system, on posterior pituitary functions is well known, but the effect of local inflammation and infection, i.e. meningitis and encephalitits, on functions of the anterior pituitary is not well documented.[br]Aims and objectives: To study the functions of anterior pituitary gland in patients suffering with meningitis and meningoencephalitis.[br]Material and Methods: Consenting patients hospitalized in the neurology services were recruited in the study. Confirmation of diagnosis was done by CSF examination and imaging, as and when required. Thyroid function tests (triiodothyronine, thyroxine and thyrotropin), sex steroids (estradiol and testosterone) and gonadotropins (LH and FSH) were collected in fasting state. Insulin induced hypoglycemia test was conducted and samples were collected for serum cortisol and growth hormone.[br]Results: Thirty three male and seventeen female patients, ranging in age from 13 - 58 years were recruited. Sixteen patients suffered from tubercular meningitis, 7 from pyogenic meningitis, 12 from viral meningitits and 15 from meningoencephalitits. Three cases had elevated TSH with low T3 while 4 had low T3, T4 and low/inappropriately normal TSH along with deficiency of other hormones. Basal cortisol, less than 14.5 microgram/dl, was seen in 29 subjects while 30 had low stimulated cortisol ([lt]18 microgram/dl). Twenty individuals had growth hormone [lt] 3.0 ng/ml on stimulation. None of the female patients had biochemical evidence of hypogonadotropic hypogonadism while 18 male subjects had low testosterone with low or inappropriately normal gonadotropins.[br]Conclusions: Abnormal thyroid profile, cortisol response and gonadal steroid status was seen in a large number of cases. Followup is required to see if it is a transient phenomenon or a long lasting one.[br][br]Sources of Research Support: Indian Council of Medical Research,. No.5/4-5/15/Neuro/2006-NCD-1(2006-04480).[br][br]Nothing to Disclose: AS, BV, CVH, BM, VS, MNR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1619 321 2575 MON-707 PO21-02 Monday 2244 2012


2240 ENDO12L_MON-708 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Opioid-Induced ACTH Deficiency or Anterior Panyhypopituitarism Sheharyar Qureshi, Ken Darzy West Middlesex University Hospital, London, UK; East and North Hertfordshire NHS Trust, London, UK Opioids are prescribed for all age groups with acute or chronic pain. The high prevalence (21% to 86%) of opioid induced hypogonadotrophic hypogonadism is under recognised and under treated. To our knowledge opioid-induced ACTH deficiency or anterior panyhypopituitarism have not been reported before. We present 2 cases of opioid-induced hypopituitarism.[br][bold]Case 1[/bold][br]A 57-year old man, with a past medical history of whiplash injury more then 12 years ago with chronic cervical pain treated with regular oral opioids. He had received Oxycontin and OxyNorm for more then 10 years. Further assessment revealed secondary hypogonadism (LH was 1 IU/l (normal range 1.5-9.3 IU/l), FSH 1.0 IU/l (2-20 IU/l), testosterone 4.1 nmol/l (8.4-18.7 nmol/l). An ITT also confirmed the presence of severe ACTH and GH deficiency. He has since been receiving hydrocortisone and testosterone replacement therapies with no evidence of recovery of the hypopituitarism more than 3 years after the diagnosis was made.[br][bold]Case 2[/bold][br]A 54-year old man, followed up in our clinic, with generalized aches and pain, tiredness, depression erectile dysfunction. He was given a diagnosis of chronic fatigue syndrome.He has been receiving regular morphine sulphate 130 mg twice per day and Oramorph up to 20 mg three times per day for chronic knee pain. His testosterone level dropped from low normal many years ago to frankly subnormal levels with inappropriately low gonadotrophins (LH 1 IU/l (normal range 2-10 IU/l), FSH 8 IU/l (1.5-33 IU/l), testosterone 3.4 nmol/l (8.4-18.7 nmol/l) and prolactin 486 pmol/l (123-1271 pmol/l). His testicular volume had reduced tremendously. He also had low free T4 and normal TSH (secondary hypothyroidism). The ITT revealed ACTH deficiency and severe GH deficiency. The pituitary MRI was normal as the rest of his biochemistry and haematology. He was treated with hydrocortisone, thyroxine and testosterone replacement therapies.[br][bold]Teaching Points[/bold][br]Patients on chronic opiate therapy should routinely be screened for possible functional hypopituitarism, which may compound the clinical picture and symptomatology.[br]For those who cannot avoid opioids, hormone replacement therapy should be offered as appropriate to relieve symptoms and prevent the long term consequences like osteoporosis.[br]Opioid-induced hypopituitarism is assumed to be functional in nature. However, we are not aware of any studies addressing the reversibility of this complication following withdrawal of opioid therapy.[br][br](1)Gilson AM et al. A reassessment of trends in the medical use and abuse of opioid analgesics and implications for diversion control: 1997-2002. J Pain Symptom Manage 2004;28:176-88. (2)Delitala G et al. The participation of hypothalamic dopamine in morphine-induced prolactin release in man. Clin Endocrinol (Oxf) 1983;19:437-44. (3)Fraser LA et al. Oral opioids for chronic noncancer pain: higher prevalence of hypogonadism in men than in women. Exp Clin Endocrinol Diabetes 2009;117:38-43. (4)Daniell HW. Opioid endocrinopathy in women consuming prescribed sustained-action opioids for control of nonmalignant pain. J Pain 2008;9:28-36.[br][br]Sources of Research Support: Possible source of funding from Lilly UK.[br][br]Nothing to Disclose: SQ, KD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 618 321 2576 MON-708 PO21-02 Monday 2245 2012


2241 ENDO12L_MON-709 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Pituitary Evaluation in Patients with Low Prostate-Specific Antigen (PSA) Mohsen Zena, Andjela Drincic, Robert Anderson, Richard Baltaro, Paul Turner VA Medical Center, Omaha, NE; Creighton University Medical Center, Omaha, NE [bold]Background[/bold]: PSA is a glycoprotein synthesized by the prostate. While the reference range for serum PSA is 0-4 ng/ml, a value [lt] 0.1 ng/dl is unusually low for any man above the age of 40. The development and growth of the prostate (and thus a PSA value) depend primarily on testosterone but, GH and IGF-1 also contribute.[br][bold]Hypothesis[/bold]: A screening PSA value below 0.1 ng/ml in otherwise asymptomatic men is associated with hypogonadism and GH deficiency.[br][bold]Results[/bold]: Hormonal evaluation was done in 49 men (ages 48-84 years) identified with a screening PSA of 0.1 ng/ml. Tests included total and free testosterone, Prolactin, LH, FSH, IGF-1, GH, TSH, FT4, AM Cortisol and ACTH. MRI of the pituitary was offered for selected patients with hypogonadotropic hypogonadism (HH) and additional hormone abnormalities. Testosterone was measured in 44 patients using the Beckman Coulter DXI Access Testosterone immunoenzymatic assay with a normal range of 170-780 ng/dl. Five patients had testosterone measured by a radioimmunoassay with a normal range of 300-890 ng/dl. We defined hypogonadism as a total testosterone value below the reference range for the assay. We found that 20 patients had low total testosterone (40.8%). Of these patients with hypogonadism, 5 had hypergonadotropic hypogonadism and 15 had hypogonadotropic hypogonadism. In 11/15 of these patients, their isolated HH was likely caused by medical comorbidities such as diabetes, obesity, narcotics and sleep apnea. Thus, they did not have a pituitary MRI. In the remaining 4 men with HH, pituitary MRI was available in one with a macroadenoma, and new profound adrenal insufficiency was diagnosed and treated. In the other 3 cases, pituitary MRI studies were done with the findings of one pituitary macroadenoma (1x1.3 cm), one pituitary microadenoma (6 mm) and one normal pituitary. In our cohort of 49 men, there were 6 patients with low IGF-1 (12.2 %): 2 patients had liver disease; 2 had pituitary macroadenomas; one had hyperprolactinemia with a normal MRI; one had normal hormone levels and no MRI was done.[br][bold]Conclusion[/bold]: Low PSA of 0.1 ng/ml or less in otherwise asymptomatic men is associated with hypogonadism in as many as 41% of patients. These men should undergo full hormonal evaluation according to the established clinical protocols to exclude other associated pituitary dysfunction.[br][br]Nothing to Disclose: MZ, AD, RA, RB, PT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1012 321 2577 MON-709 PO21-02 Monday 2246 2012


2242 ENDO12L_MON-710 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Clinical Presentation and Bone Density in Sheehan Syndrome in North India Subhash Yadav, Ramesh Gomez, Eesh Bhatia Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, India [bold]Introduction:[/bold][br]Sheehan[apos]s syndrome results from pituitary insufficiency due to excessive blood loss during or after delivery. Insufficient antenatal care, high frequency of home delivery, and co-morbid conditions like anemia make Sheehan[apos]s syndrome a leading cause of pan hypopituitarism in India. Delay in diagnosis and treatment, poor nutrition and low body mass may predispose Indian patients to severe and early osteoporosis.[br][bold]Aims and Objectives:[/bold][br]To study the demographic, clinical and hormonal profile of patients with Sheehan[apos]s syndrome and to determine the prevalence of osteoporosis.[br][bold]Study design: [/bold]Cross sectional study of 35 patients with Sheehan[apos]s syndrome presenting to the Endocrine clinic at a teaching hospital between 2005 and 2011.[br][bold]Results: [/bold]Patients had mean age at diagnosis of 40.2[plusmn]10 years (range 24-53 years) and interval between disease onset and diagnosis 9.4 [plusmn] 6.9 years (range 2 weeks-29 years). Seventy one percent came from an urban area; 79% had delivery at primary health care or community centre hospital. Two- thirds of patients came from low or low middle income groups. The most common presentation were secondary amenorrhea, lactation failure and weight loss (94%, 97% and 97% respectively). Adrenal crisis was the presentation in 50%, 17% had acute confessional state. Forty eight percent of the patients had moderate to severe anemia. Abnormal lipid profile was frequent (elevated triglycerides 65%, elevated LDL cholesterol 57% and low HDL cholesterol 35%). All patients had central hypothyroidism and hypogonadism, and 97% had hypocortisolism. On MRI 70% had empty sella and 20% had partial empty sella. Osteoporosis was present in 26% (22% at lumbar spine and 16% at hip). Osteopenia was detected in 74% (74% at lumbar spine and hip). Osteoporosis present in 29% of the patients below 40 years. Age (r=-0.39, p=0.04) and serum alkaline phosphatase (r=-0.5, p=0.05) showed significant correlation with bone mineral density. No patients had any radiological evidence of fractures.[br][bold]Conclusion: [/bold]Post-partum pituitary necrosis had a delayed and severe presentation in this series. Osteoporosis at both spine and hip were common in Sheehan[apos]s syndrome. Increased awareness, early diagnosis and replacement may improve outcomes.[br][br](1) Tessnow AH et al., Am J Med Sci. 2010;340:402. (2) Gokalp D et al., Gynecol Endocrinol. 2009;344:349.[br][br]Sources of Research Support: Intramural Research grant by SGPGI (PGI/DIR/RC/28/2012).[br][br]Nothing to Disclose: SY, RG, EB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1085 321 2578 MON-710 PO21-02 Monday 2247 2012


2243 ENDO12L_MON-711 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Increased Prevalence of Cardiovascular Risk Factors and Metabolic Syndrome in Hypopituitary Patients Yoon Ji Kim, Won Sang Yoo, Ah Reum Khang, Jung Hee Kim, Sang Wan Kim, Seong Yeon Kim Seoul National University College of Medicine, Seoul, Korea Previous reports suggested that the incidence of cardiovascular disease and the prevalence of dyslipidemia are higher in hypopituitary patients. The aim of present study was to estimate the prevalence of cardiovascular risk factors and metabolic syndrome in hypopituitary patients and to investigate the differences in prevalence of cardiovascular risk factors and metabolic syndrome between hypothalamic and pituitary patients.[br]Eight hundred fifty five medical records of hypopituitarism (pituitary/hypothalamic lesion) from 2000 to 2010 were evaluated and all of them were followed up in Seoul national university hospital. Five hundred forty patients are included, who were on hormone replacement treatment, above 20 years old and had 1 year or above of follow up duration. The exclusion criteria were hypopituitarism due to congenital or inflammatory disease, patients who had only chemotherapy and cancer metastasis.[br]Mean age at time of diagnosis of hypopituitarism was 36.0 [plusmn] 18.4 years. Of the 540 patients, 298 were male (55.2%). They were followed up for 9.9 [plusmn] 6.8 years and on a replacement therapy for average of 2.5 hormones. The prevalence of diabetes mellitus (fasting blood glucose [ge] 126 mg/dL, 10 vs 15.2%, P [lt] 0.05), overweight and obesity (body mass index [ge] 25 kg/m2, 33.1 vs 53.9%, P [lt] 0.05), hypercholesterolemia (total cholesterol [ge] 240 mg/dL, 11.2 vs 30%, P [lt] 0.05), hypertriglyceridemia (triglyceride [ge] 200 mg/dL, 17.4 vs 34.8%, P [lt] 0.05), high LDL cholesterol (LDL [ge] 130 mg/dL, 24.9 vs 32.5%, P [lt]0.05) and metabolic syndrome (31.3 vs 42.3%, P [lt] 0.05) was higher in hypopituitary patients than the Korean National Health and Nutrition Examination Surveys (2008 KNHANES) population. The prevalence of high LDL cholesterol in hypothalamic lesion group (208/540, 38.3%) was higher than pituitary lesion group (332/540, 61.7%) adjusting for age, sex, treatment modalities (operation/radiotherapy/operation and radiotherapy) and follow-up duration. The prevalence of metabolic syndrome was higher in hypothalamic lesion group than pituitary lesion group in the female patients older than fifty years.[br]The prevalence of cardiovascular risk factors and metabolic syndrome was higher in hypopituitary patients than general Korean population. The results of present study suggest that patients with hypothalamic lesions have higher prevalence of cardiovascular risk factors and metabolic syndrome compared to patients with pituitary lesion.[br][br]Nothing to Disclose: YJK, WSY, ARK, JHK, SWK, SYK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 743 321 2579 MON-711 PO21-02 Monday 2248 2012


2244 ENDO12L_MON-712 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Diabetes Insipidus Could Be the Initial Sign of Primary Hypophysitis Se Won Kim, Sun-Mi Park, Hye Jeong Kim, Yoon Young Cho, Ji Cheol Bae, Jae Hoon Chung, Sun Wook Kim, Jae Hyeon Kim, Kwang-Won Kim Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea [bold]Introduction[/bold][br]The natural history of primary hypophysitis is incompletely understood and best treatment remains controversial.[br][bold]Methods[/bold][br]We performed a retrospective study of 19 patients (mean age of 47 years, fifteen women and four men) who were diagnosed with primary hypophysitis in Samsung Medical Center in Seoul, Korea, from January 2001 to October 2011.[br][bold]Results[/bold][br]Two patients had recent pregnancy and underlying autoimmunity. Most common clinical feature was polyuria and polydipsia (78%) and most common endocrinological finding was also diabetes insipidus (DI, 73%). The initial presumptive diagnosis was pituitary adenoma or lymphoma in eight patients (42%) and inflammatory hypophysitis in eleven (57%). The first eight patients underwent transsphenoidal surgery but only four (21%) proceeded to total tumor removal. Four patients (21%) received steroid therapy of whom one patient relapsed with pressure symptoms and later underwent surgery. Half of the patients under steroid treatment suffered from side effects such as facial swelling and body weight gain. Only one patient (25%) did not respond satisfactorily to steroid therapy and later underwent surgery. Altogether, the histological diagnoses were confirmed in nine patients, seven lymphocytic hypophysitis (77%), each one of granulomatous and xanthomatous hypophysitis (11%, respectively). All patients who underwent surgery required long-term hormone replacement, three (75%) relapsed and needed additional radiotherapy or steroid therapy. Eleven (57%) patients who had no pressure or visual symptoms received only conservative management with hormone replacement (i.e. vasopressin for DI). All these eleven patients have been regularly followed up without recurrence.[br][bold]Conclusion[/bold][br]Diabetes insipidus could be the initial sign of primary hypophysitis and controlled satisfactorily without aggressive treatment.[br][br]Nothing to Disclose: SWK, S-MP, HJK, YYC, JCB, JHC, SWK, JHK, K-WK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1041 321 2580 MON-712 PO21-02 Monday 2249 2012


2245 ENDO12L_MON-713 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Hypernatremia Is More Common Than Hyponatremia Following Acute Traumatic Brain Injury and Is Associated with Higher Mortality Mark J Hannon, Rachel K Crowley, Lucy Ann Behan, Eoin P O[apos]Sullivan, Eoin De Faoite, Michelle M O[apos]Brien, Karen Duggan, Danny Rawluk, Rory O[apos]Dwyer, Amar Agha, Christopher J Thompson Beaumont Hospital/RCSI Medical School, Dublin, Ireland; Beaumont Hospital/RCSI Medical School, Dublin, Ireland; Beaumont Hospital/RCSI Medical School, Dublin, Ireland Published data have shown that hypernatraemia, which is usually due to acute cranial diabetes insipidus (DI), is common following acute traumatic brain injury (TBI) (1). Hyponatraemia may occur following any neurosurgical insult and may be due to the Syndrome of Inappropriate AntiDiuresis (SIAD), acute glucocorticoid deficiency or cerebral salt wasting syndrome (CSWS) (2). We performed a prospective study to determine the precise incidence and causes of hypernatraemia and hyponatraemia in acute TBI and the relationship of these electrolyte imbalances to patient mortality.[br]100 patients (84 men, median age 33, range 18 to 75), with acute TBI (mean GCS +/- SD = 8.59 +/- 4.2) were prospectively recruited. Each patient had their electrolytes measured and fluid balance recorded daily for 10 consecutive days following TBI. If any electrolyte imbalance was detected, urinary sodium and osmolality were also measured. The presence of hypotonic ([lt] 300 mOsm/kg) polyuria ([gt] 2 ml/kg/hr) with plasma sodium [gt] 145mmol/L was regarded as diagnostic of DI (3). SIAD was defined according to standard diagnostic criteria (4). Plasma cortisol was measured at 0900h on days 1, 3, 5, 7 and 10 following TBI; a plasma cortisol [lt] 300 nmol/L in a patient in ITU was regarded clinically as inappropriately low and suggestive of acute glucocorticoid deficiency.[br]55/100 developed hypernatraemia (plasma sodium [gt] 145 mmol/L) due to diabetes insipidus, 15 patients developed hyponatraemia (plasma sodium [lt] 135 mmol/L), and 4 developed hyponatraemia followed by hypernatraemia. Mortality was significantly higher in those with DI when compared with those with hyponatraemia (p=0.027) and those with normal plasma sodium (p=0.02), although there were no differences in length of hospital stay or severity of TBI. The median time of onset of hypernatraemia was 6 days (range 1-9); in those with hyponatraemia it was 3 days (range 1-9). 44/55 patients with hypernatraemia and 13/15 patients with hyponatraemia had acute cortisol deficiency. Those patients with hyponatraemia and hypocortisolaemia had normalisation of their plasma sodium following glucocorticoid treatment; the other 2 had SIAD which responded to fluid restriction. There were no cases of CSWS.[br]Hypernatraemia in acute TBI is more common than previously reported and is associated with a higher mortality than hyponatraemia, and is almost exclusively due to acute cranial DI. Hyponatraemia in TBI is due to SIAD and acute glucocorticoid deficiency.[br][br]1. Agha A, Sherlock M, Phillips J, Tormey W, Thompson CJ. The natural history of post-traumatic neurohypophysial dysfunction. Eur J Endocrinol. 2005 Mar;152(3):371-7. 2. Sherlock M, O[apos]Sullivan E, Agha A, Behan LA, Owens D, Finucane F, et al. Incidence and pathophysiology of severe hyponatraemia in neurosurgical patients. Postgrad Med J. 2009 Apr;85(1002):171-5. 3. Seckl J, Dunger D. Postoperative diabetes insipidus. BMJ. 1989 Jan 7;298(6665):2-3. 4. Verbalis JG. Hyponatraemia. Balliere[apos]s Clinical Endocrinology and Metabolism. 1989 1989;3:499-530.[br][br]Nothing to Disclose: MJH, RKC, LAB, EPO, EDF, MMO, KD, DR, RO, AA, CJT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1966 321 2581 MON-713 PO21-02 Monday 2250 2012


2246 ENDO12L_MON-714 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Hyponatremia Assessment and Outcomes in Acute Medically Ill Patients Edward Jude, Anna Hughes, Omer Taha, Tony Tetlow Tameside Hospital NHS Foundation Trust, Ashton under Lyne, UK Background: Hyponatremia is the most common electrolyte abnormality, encountered in up to 30% of inpatients. Inappropriate management can have serious implications for patients; including demyelinating disease, coma, and death.[br]Methods: Patients (n=122) admitted to the medical admission unit of a district general hospital with a serum sodium (Na) [le]130 were selected for the study. All details including patient demographics, blood biochemistry, date of admission and date of death were taken from the case notes and hospital computerised system. Details on assessment of hyponatremia including thyroid, adrenal and renal function were also recorded.[br]Results: Mean age was 70.4+18.1 years; 48 males. Mean serum Na on admission was 125.8+4.1 mmol/l. Of the 122 patients, 38 (31.1%) died in hospital. Patients who died were older (66.7[plusmn]18.9 vs 72.3[plusmn]15.3; p=0.054) Admission serum Na and plasma glucose in survivors vs died was 127.2[plusmn]3.9 vs 124.2[plusmn]4.7 mmol/l (p[lt]0.001); and 7.0[plusmn]3.0 vs 5.4[plusmn]2.2 mmol/l (p=0.02) respectively. Patients with admission serum Na [le] 125 in the survivors vs died was 32.1 vs 66.7%. Patients investigated for hyponatremia were as follows: serum cortisol (n= 6), plasma osmolality (n=9), urine osmolality (n= 9), short synacthen test (n=0), urine sodium (n=3), thyroid function tests (n=19). Conclusions: Patients admitted with acute medical conditions with severe hyponatremia have a high mortality and those with lower serum sodium have greater risk of death. Patients were also inadequately worked up for assessment of cause of the hyponatremia and further education of medical specialists is urgently required to improve management and outcome. Also lower admission plasma glucose was associated with higher mortality.[br][br]Nothing to Disclose: EJ, AH, OT, TT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2077 321 2582 MON-714 PO21-02 Monday 2251 2012


2247 ENDO12L_MON-715 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Tolvaptan in Clinical Practice: A Single Center Experience from United Kingdom Rajesh Rajendran, Partha Kar Queen Alexandra Hospital, Portsmouth, UK [bold]Introduction:[/bold]Hyponatremia(serum sodium concentration[lt]135mEq/L) is the most common electrolyte disorder in hospitalized patients,affecting up to 30% of inpatients(1).A number of treatment modalities have been reviewed with no consensus on the most optimal approach(2,3).Two multicenter randomized controlled trials showed that tolvaptan is effective and safe in the treatment of euvolemic hyponatremia(4).We report the first consecutive retrospective clinical case series from a single center in United Kingdom.[br][bold]Aim:[/bold]We analyzed the safety and efficacy of tolvaptan in the treatment of hyponatremia in hospitalized inpatients.[br][bold]Methods:[/bold]All subjects treated with tolvaptan for hyponatremia were included in this study,except one subject whose case notes could not be traced.15 episodes of treatment with tolvaptan in 14 subjects were analyzed.[br][bold]Results:[/bold]There were 8 women and 6 men(mean age 72[plusmn]16,36 to 90years and mean BMI 24.9[plusmn]8.67,13.9 to 46.4kg/m[sup]2[/sup]).Confusion was present on admission in all 15 episodes,with falls and mood changes reported in 11/15 episodes.13 subjects were diagnosed with euvolemic hyponatremia based on fluid status,serum and urine osmolalities and urinary sodium excretion.One subject had hypotonic hyponatremia.The duration of hyponatremia was unknown in 8,chronic([gt]48 hours) in 4 and acute([le]48 hours) in 3 episodes.During 12 episodes,subjects were fluid restricted to [le]1 litre(mean duration 5.91[plusmn]3.11,2 to 12 days),prior to commencement of tolvaptan.The median duration of tolvaptan therapy was 3 days(1 to 21 days).Sodium level of 130mEq/L was targeted during therapy and fluid restriction was discontinued.There was a significant change in sodium levels between commencement(mean sodium 120.07[plusmn]4.59,108 to 126mEq/L) and end(mean sodium 131.87[plusmn]3.58,125 to 139mEq/L,p[lt]0.0001) of tolvaptan therapy.Maximum rate of change of sodium was observed in the first 24 hours of therapy(mean 6.73[plusmn]2.84,1 to 11mEq/L) with no subject exceeding 12mEq/L in 24 hours and 18mEq/L in 48 hours at any point whilst on tolvaptan,above which osmotic demyelination syndrome can occur(5).There were small insignificant rises in serum urea(p=0.4843) and creatinine(p=0.8410),between start and end of tolvaptan therapy.One subject complained of thirst during therapy and no subject developed osmotic demyelination syndrome.[br][bold]Conclusions:[/bold]Tolvaptan,a selective oral vasopressin V2-receptor antagonist appears to be safe and efficacious in the management of hospitalized inpatients with euvolemic hyponatremia.[br][br](1) Upadhyay A et al., Incidence and prevalence of hyponatremia. Am J Med 2006;119(suppl 1):S30-S35. (2) Ellison DH et al., The syndrome of inappropriate antidiuresis. N Eng J Med 2007;356:2064-72. (3) Verbalis JG et al., Hyponatremia Treatment Guidelines 2007: Expert Panel Recommendations. Am J Med 2007;120(11A):S1-S21. (4) Schrier RW et al., Tolvaptan, a selective oral vasopressin V2-receptor antagonist for hyponatremia. N Eng J Med 2006;355:2099-2112. (5) Sterns et al., Neurologic sequelae after treatment of severe hyponatremia: a multicenter perspective. J Am Soc Nephrol. 1994;4:1552-1530.[br][br]Nothing to Disclose: RR, PK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 262 321 2583 MON-715 PO21-02 Monday 2252 2012


2248 ENDO12L_MON-716 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Epidemiology and Presentation of Pituitary Tumors in Iceland 1955[ndash]2010 Tomas Thor Agustsson, Tinna Baldvinsdottir, Paul Carroll, Barbara McGowan, Rafn Benediktsson Lanspitali University Hospital, Reykjavik, Iceland; Guy[apos]s and St Thomas[apos] NHS Foundation Trust, London, UK Background[br]Autopsy and imaging studies suggest that pituitary tumours are amongst the most common human neoplasm. Few population-based studies have assessed the epidemiology and presentations of clinically significant pituitary tumours.[br]Iceland has a small well defined population. We have kept a database of all pituitary tumours diagnosed in Iceland from 1955 until 2010.[br]The aim of this study is to describe the epidemiology of pituitary tumours in Iceland[br]Methods[br]This is a retrospective observational study The cohort was divided into three equal sized groups by date of diagnosis. We also mapped the home of each patient on a map of Iceland. To assess their presentation we examined sex ratios, type and size of tumour, presence and nature of symptoms and related that to peroid of diagnosis.[br]Results[br]We identified a total of 392 individuals, 165 men (42%) and 227 women (58%). 320 are alive, giving a current prevalence of exactly 0.1%. Median age at diagnosis was 44.4. 43% had a non-functioning adenoma, 40.1% had a prolactinoma, 11.2% had acromegaly, and 5.6% had Cushing[apos]s disease.[br]Acromegaly and Cushing[apos]s disease presented almost exclusively with symptoms associated with hormonal hypersecretion. Non-functioning adenomas were more likely to be incidental, associated with localized symptoms, and present at an older age.[br]Macroadenomas presented at an older age than microadenomas, were more often non-functioning, and associated with localized symptoms rather than hormonal dysfunction.[br]Women presented at a younger age than men and were less likely to present with a macroadenoma or localized symptoms.[br]Comparing the three time periods there was no significant change in the incidence of macroadenomas, but non-functioning tumours and incidentally discovered tumours were more common later. Patients in the first group were younger at diagnosis[br]Geographical mapping showed aggregation towards more densely populated areas possibly with better access to imaging.[br]Conclusion[br]The epidemiology and characteristics of pituitary tumours in Iceland is similar to other studies. The incidence of pituitary tumours is rising. This may be related to greater access to imaging and earlier diagnosis. However, in this study, more recently diagnosed patients did not present younger or have smaller tumours. Clinically significant pituitary tumours are apparently becoming more prevalent which does emphasize the importance of raising awareness of this condition and doing further research in this area.[br][br]Nothing to Disclose: TTA, TB, PC, BM, RB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2304 322 2584 MON-716 PO22-03 Monday 2253 2012


2249 ENDO12L_MON-717 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) [quot]Epiacro[quot]: The Mexican Acromegaly Registry Paola Jervis, Cesar Trujillo, Lesly Portocarrero, Benito Dominguez, Maricela Vidrio, Rosa Isela Luna-Ceballos, Alfredo Reza, Francisco Velazquez, Maria Lourdes Maya-Luna, Graciela Gomez, Karina Acevedo, Avril Garcia, Moises Mercado Centro Medico Nacional, S XXI, Instituto Mexicano del Seguro Social, Mexico City, Mexico; Sociedad Mexicana de Nutricion y Endocrinologia, Mexico City, Mexico; Novartis, Mexico City, Mexico Background: Acromegaly is a disease of low prevalence, with an incompletely defined epidemiology.[br]Objective: To inform the first set of results of the National Acromegaly Registry, or [ldquo]Epiacro[rdquo] program.[br]Design and methodology: The data base included clinical, biochemical, imaging, therapeutic and outcome information. Fifteen centres spread through out the Country participated in the study, registering patients diagnosed after 1990.[br]Results: To date, 1328 patients have been registered, of whom 60% are women. Mean age is 41.6 [plusmn] 12 years. Global prevalence was 13 cases per million, whereas the regional prevalence ranged from 7 to 49 cases per million. The most commonly reported symptoms were acral enlargement, headache, arthralgia, fatigue, hyperhydrosis and snoring. In almost half of the patients, an abnormality in glucose metabolism was found, one third had frank diabetes and another third had hypertension. In over 80% of cases a basal GH level was available, whereas in close to 60% the biochemical diagosis was based on either a post-glucose GH and/or an IGF-1 level. In 70%, imaging studies revealed a non-invasive macroadenoma. Pituitary surgery was the most frequently used treatment, and over 40% required pharmacological treatment, usually with somatostatin analogs.[br]Conclusion: In this first report of [ldquo]Epiacro[rdquo] we found a slightly lower disease prevalence than that reported in other parts of the World, a fact that highlights an important degree of underdiagnosis.[br][br]Sources of Research Support: Novartis Oncology, Mexico.[br][br]Disclosures: CT: Employee, Novartis Pharmaceuticals. Nothing to Disclose: PJ, LP, BD, MV, RIL-C, AR, FV, MLM-L, GG, KA, AG, MM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 79 322 2585 MON-717 PO22-03 Monday 2254 2012


876 ENDO12L_SUN-26 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Male First-Degree Relatives of Women with Polycystic Ovary Syndrome Have Defects in Insulin Action and Energy Expenditure Sudha K Yalamanchi, Leanne Redman, Andrea Dunaif Feinberg School of Medicine, Northwestern University, Chicago, IL; Pennington Biomedical Research Center, Baton Rouge, LA Polycystic ovary syndrome is a highly heritable complex genetic disease. Male as well as female first-degree relatives (FDRs) have reproductive and metabolic phenotypes. In male FDRs, the metabolic phenotypes include insulin resistance and evidence for [beta]-cell dysfunction. We performed this study to further assess insulin action and secretion as well as substrate oxidation by indirect calorimetry.[br]Fourteen non-Hispanic White male FDRs and 19 control (CON) of comparable age, BMI and ethnicity had graded glucose infusion and 120 min 80 mU/m2/min ([sim]200 [mu]U/mL steady-state insulin) euglycemic clamp studies. Body composition was assessed by DEXA and visceral adipose tissue (VAT) by MRI. Indirect calorimetry was performed under basal conditions and during the last 45 minutes (steady-state period) of the clamp.[br]Fat mass, fat free mass (FFM) and VAT did not differ in between groups. Insulin-mediated glucose disposal (IMGD) was significantly lower in FDRs (7.18 [plusmn] 3.17 FDR v 9.38 [plusmn] 2.49 CON mg/kg FFM/min, P= 0.018). Insulin secretion rates were similar at all doses during graded glucose infusion. The disposition index calculated as the product of IMGD and maximal insulin secretion rate did not differ between groups. Baseline and insulin-stimulated respiratory quotients (RQ) did not differ but the change (insulin-mediated minus baseline) in RQ was lower in FDRs (0.05 [plusmn] 0.04 FDR v 0.09 [plusmn] 0.05 CON, P= 0.027). Basal, insulin-mediated and the change in glucose oxidation did not differ between groups. Non-oxidative glucose disposal was significantly lower in FDRs (6.93 [plusmn] 3.11 FDR v 9.14 [plusmn] 2.50 CON mg/kg FFM/min, P= 0.0167). Basal and insulin-stimulated fat oxidation did not differ but the change in fat oxidation was significantly lower in FDRs (-0.027 [plusmn] 0.018 FDR v -0.041 [plusmn] 0.021 CON, P= 0.048).[br]Male FDRs had decreased IMGD without evidence for defects in insulin secretion. Consistent with a previous report in male FDRs and findings in other insulin resistant groups, only non-oxidative glucose disposal was impaired. FDRs had metabolic inflexibility with a reduced change in RQ, perhaps due to the reduction in IMGD. Suppression of fat oxidation by insulin was also impaired in FDRs. This abnormality has been hypothesized to be an early risk marker for type 2 diabetes, independent of insulin secretion and IMGD. In conclusion, insulin resistance in male FDRs affects primarily non-oxidative glucose metabolism as well as fat oxidation.[br][br]Nothing to Disclose: SKY, LR, AD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1625 174 1083 SUN-26 PO34-01 Sunday 878 2012


877 ENDO12L_SUN-27 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Birth Weight and PCOS in Adult Life. A Register-Based Study on 523,757 Danish Women Born 1973-1991 Hanne Mumm, Mads Kamper-Jorgensen, Anne-Marie Nybo Andersen, Glintborg Dorte, Andersen Marianne Odense University Hospital, Odense, Denmark; University of Public Health, Copenhagen, Denmark [bold]Objective: [/bold]To study the association between birth weight and polycystic ovary syndrome (PCOS) in adult life in Danish women born 1973-1991.[br][bold]Methods: [/bold]All female children born of Danish mothers in Denmark during 1973-1991 were included (n = 523,757) and followed for a total of 4,739,547 person-years at risk. Information on birth weight was extracted from the Danish Medical Birth Register. The cohort was followed-up for hospitalisations with the diagnoses hirsutism or PCOS from age 15 years until the end of 2006 in the Danish National Patient Register (NPR). Information on maternal diabetes diagnoses was also extracted from the NPR. We estimated incidence rate ratios (IRR) from Poisson regression models with 95 % confidence intervals (CI). All analyses were adjusted for attained age and period.[br][bold]Results:[/bold] The risk of PCOS was significantly increased in women with birth weight [gt] 4,500 grams (IRR = 1.57 (95 % CI 1.21-2.03)) compared to women with birth weight 3,000-3,499 grams. The risk of PCOS was independent of size for gestational age. Offspring of mothers diagnosed with diabetes were at increased risk of PCOS. In these women the risk of PCOS increased with decreasing fetal growth.[br][bold]Conclusions:[/bold] Our data suggest that the risk of PCOS was increased in girls with birth weight [gt] 4,500 grams. In offspring of diabetic mothers we found an inverse relation between fetal growth and PCOS.[br][br]Nothing to Disclose: HM, MK-J, A-MNA, GD, AM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1260 174 1084 SUN-27 PO34-01 Sunday 879 2012


878 ENDO12L_SUN-28 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Reproductive Features after Menarche in Daughters of Women with Polycystic Ovary Syndrome Nicolas Crisosto, Barbara Echiburu, Amanda Ladron de Guevara, Jessica Preissler, Manuel Maliqueo, Teresa Sir-Petermann University of Chile, Santiago, Chile [bold]Context[/bold]: Polycystic ovary syndrome (PCOS) is a common familial endocrine-metabolic disorder. It has been proposed that an important proportion of the first-degree female relatives of PCOS patients may be at risk for developing PCOS. In a recent study we observed that 45% of PCOS daughters (PCOSd) are affected in Tanner IV and 60% in Tanner V, taking biochemical hyperandrogenism as the main criterion. However, it is not known how many PCOSd ultimately develop PCOS in the early reproductive period. [bold]Objective[/bold]: To determine how many PCOSd exhibit a PCOS according to the AE-PCOS Society criteria after menarche. [bold]Design[/bold]: Forty eight daughters of PCOS women (PCOSd) and 27 daughters of control women (Cd) were studied 1, 2 and 3 years after menarche. In both groups, a GnRH agonist test (leuprolide acetate, 10 ug/Kg s.c.) with measurement of circulating concentrations of gonadotropins, sex steroids, sex hormone binding globulin (SHBG) and anti- M[uuml]llerian hormone (AMH) was performed. Concomitantly, ovarian volumes were determined by ultrasound[bold]. [/bold][bold]Results[/bold]: Both groups had similar age of menarche(11.7 PCOSd and 11.6 Cd), BMI SDS, and waist to hip ratio. Gynecological age was 2.0 for PCOSd and 1.6 for Cd. Basal and stimulated LH and FSH concentrations were similar between both groups one, two and three years after menarche. Basal testosterone and AMH concentrations were always higher in the PCOSd group compared to the Cd group during the three year study period. On the whole, 69% of PCOSd and 14% of Cd showed hyperandrogenism. 31% of PCOSd and 14% of Cd showed irregular menstrual cycles and a polycystic ovarian morphology. Finally, 33% of PCOSd and 14% of Cd met the criteria for PCOS according to AE-PCOS Society. Interestingly, only 36% of the PCOSd that fulfilled the criteria for PCOS had increased AMH concentrations. [bold]Conclusions[/bold]: Our results indicate that hyperandrogenism is the main feature for the diagnosis of PCOS during the postmenarchal period as previously suggested. Menstrual irregularity, PCO morphology and AMH levels are secondary elements that may aid in the diagnosis of this syndrome.[br][br]Sources of Research Support: FONDECYT Grant 1071007.[br][br]Nothing to Disclose: NC, BE, ALdG, JP, MM, TS-P 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 91 174 1085 SUN-28 PO34-01 Sunday 880 2012


879 ENDO12L_SUN-29 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) A Novel Method for Metabolic Classification of Polycystic Ovarian Syndrome Evgenia A Gourgari, Chris Crutchfield, Matthew Olson, Alfred Yergey, Constantine Stratakis National Institute of Child Health and Human Development, Bethesda, MD; National Institute of Child Health and Human Development, Bethesda, MD; John Hopkins Hospital, Baltimore, MD Polycystic ovarian syndrome (PCOS) is a heterogeneous group of disorders associated with hyperandrogenism. Criteria set by the Androgen Excess and PCOS Society for the diagnosis of PCOS stress hyperandrogenism with either menstrual irregularity and/or polycystic ovaries on ultrasound. The etiology of PCOS remains unknown, despite its links to insulin resistance, hypertension and metabolic syndrome. Corticotropin (ACTH)-independent or ACTH-dependent functional adrenal hyperandrogenism may accompany functional ovarian hyperandrogenism; rarely it accounts solely for hyperandrogenemia in PCOS. Our goal was to develop a new method that could detect with great specificity androgen metabolites of adrenal and ovarian origin in women with PCOS and thus lead to better understanding of the pathophysiology and clinical manifestations of the disease. We have developed a novel method combining data visualization and mass spectrometry (MS) to compare concentrations of urinary steroids and used it in a study comparing patients with polycystic ovarian syndrome (PCOS) and healthy volunteers (HV). This method enables an unbiased quantitative evaluation of steroids that may have not been previously assayed using a traditional ELISA-based approach, or that may have had their measurement confounded by cross-reactivity with a molecule of similar structure. Apilot analysis comparing PCOS (n=5) and HV (n=4) revealed several analytes that were consistently increased in PCOS. We focused on 2 features exhibiting the greatest magnitude changes. The first matched MS and chromatographic characteristics (LC) of dehydroepiandrosterone sulfate (DHEA-S). The second feature matched MS characteristics but not LC for dihydrotestosterone. We speculate that this molecule is [alpha]-dihydrostestosterone-sulfate (DHT-S). Quantitative comparison of these two analytes were made relative to internal standards and normalized by creatinine. Averages as means +/- 1 standard deviation and P values (2-tailed t-test assuming equal variance) yielded: DHEA-S: HV mean= 8.5+/-9.8, PCOS mean= 119.9+/-57.8, fold-change=12.4, p-value=0.015. DHT-S: HV average= 4.1+/-1.5, PCOS Average=18.3+/-4.4, fold-change=4.5, p-value= 0.00047. The cohort of patients is actively and prospectively recruited. We conclude that this method provides sufficient molecular specificity to enable metabolic sub-classification of hyperandrogenism in patients with PCOS and should determine the degree of adrenal steroid contribution.[br][br]*Drs. Gourgari and Crutchfield have contributed equally to this work.[br][br]Nothing to Disclose: EAG, CC, MO, AY, CS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1313 174 1086 SUN-29 PO34-01 Sunday 881 2012


880 ENDO12L_SUN-30 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Younger Age at Menarche Is Associated with a More Hyperandrogenic Phenotype and Adverse Metabolic Profile in Patients with PCOS Yvonne V Louwers, Samantha M Verburg, Ramon HM Dykgraaf, Joop SE Laven Erasmus Medical Center, University Medical Center, Rotterdam, Netherlands [bold]Background [/bold]Polycystic Ovary Syndrome (PCOS) is the most common endocrinopathy in women of reproductive age and its associated co-morbidities include insulin resistance, metabolic syndrome and cardiovascular disease. Identification of risk groups within the broad PCOS phenotype is essential in screening and prevention of these long-term health risks. In the general population young age of menarche, i.e., below 12 years of age, is a known predictor for adverse metabolic profile. The aim of our study was to determine whether age at menarche influences the phenotype and components of the metabolic syndrome in PCOS patients.[br][bold]Material and methods[/bold] The study population consisted of 1879 patients diagnosed with PCOS according to the 2003 Rotterdam consensus criteria. The patients were divided into three groups based on age at menarche: [lt] 12 years, 12-14 years and [gt] 14 years. Regression analyses were performed to assess differences in the phenotype of PCOS and components of the metabolic between the menarchal age groups. Analyses were adjusted for age at screening, BMI, ethnicity and post-menarchal age, i.e., interval between age at menarche and screening.[br][bold]Results[/bold] The majority of the patients (n=1207, 64.2%) was between 12 and 14 years of age when menarche occurred, whereas 297 (15.8%) patients were under the age of 12 years and 375 (19.9%) patients were older than 14 years. PCOS patients with age at menarche under 12 years had a significantly higher prevalence of hyperandrogenism compared to the other menarchal age groups (71% in [lt] 12 yrs; 59% in 12-14 yrs; 48% in [gt] 14 yrs; p-value [lt]0.001). The full-blown phenotype (hyperandrogenism, ovulatory dysfunction and polycystic ovaries) was significantly more often present in this age of menarche group (59% in [lt] 12 yrs; 49% in 12-14 yrs; 40% [gt] 14 yrs; p-value [lt] 0.001). Furthermore, patients who had their menarche before the age of 12 years were more often obese (42% in [lt] 12 yrs; 30% in 12-14 yrs; 22% [gt] 14 yrs; p-value [lt] 0.001) and had more often a waist circumference [gt] 102 cm (22% in [lt] 12 yrs; 16% in 12-14 yrs; 12% [gt] 14 yrs, p-value [lt]0.05).[br][bold]Conclusion [/bold]Early menarche is associated with an more hyperandrogenic phenotype of PCOS and with a higher prevalence of components of the metabolic syndrome.[br][br]Sources of Research Support: J.S.E.L. has received fees and grant support from the following companies: Ferring, Genovum, Merck-Serono, Organon, Schering Plough and Serono.[br][br]Nothing to Disclose: YVL, SMV, RHMD, JSEL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 792 174 1087 SUN-30 PO34-01 Sunday 882 2012


881 ENDO12L_SUN-31 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Developmental Programming: Prenatal and Postnatal Contribution of Androgen and Insulin in the Reprogramming of Pubertal Onset in Prenatal Testosterone-Treated Sheep Almudena Veiga-Lopez, Bachir Abi Salloum, Alexandria Williams, Carol Herkimer, Evan Beckett, Vasantha Padmanabhan University of Michigan, Ann Arbor, MI; University of Michigan, Ann Arbor Prenatal testosterone (T) excess advances neuroendocrine puberty and leads to reproductive defects (oligo-anovulation, functional hyperandrogenism, and multifollicular ovaries) similar to that seen in women with polycystic ovary syndrome. These animals also develop metabolic disruptions such as insulin resistance. The objective of this study was to determine the organizational and activational role of androgen and insulin in programming pubertal advancement. The study tested the hypothesis that antagonizing androgen action or improving insulin sensitivity prenatally would prevent pubertal advancement while postnatal treatment with androgen antagonist or insulin sensitizer would reduce this advancement. The following groups were studied: 1) control, 2) prenatal T (100 mg T propionate twice weekly from days 30-90 of gestation, i.m.), 3) prenatal T plus 15 mg/kg/day androgen antagonist, flutamide s.c. (TF), 4) prenatal T plus 8 mg/day insulin sensitizer, rosiglitazone orally (TR), 5) prenatal T and postnatal androgen antagonist (T+F), 6) prenatal T and postnatal insulin sensitizer (T+R) (n=6-30 animals/group). Postnatal treatment began at [sim]6 weeks of age. Twice weekly blood samples were taken beginning at [sim]12 weeks of age to determine the timing of onset of progestogenic cycles. Data were analyzed by analyses of variance. One T female and one TF female did not enter puberty and were excluded from analyses. Control females entered puberty at 27.5[plusmn]0.34 weeks of age ([sim]October 15[sup]th[/sup]). Prenatal T females entered puberty earlier (P[lt]0.05) (22.5[plusmn]2.5 wks; [sim]September 10[sup]th[/sup]) compared to controls. Pubertal timing of the intervention groups did not differ from that of controls (TR: 26.5[plusmn]1.0; TF: 25.4[plusmn]0.7; T+R: 27.7[plusmn]0.7; T+F: 28.8[plusmn]0.9 weeks of age). These findings indicate that prenatal T not only advances neuroendocrine puberty, as reported in the prenatally ovariectomized model, but also the start of the progestogenic cycles. Rescue of the phenotype by co-treatment with an androgen antagonist or an insulin sensitizer suggests that this advancement is due to androgenic programming and involves insulin as a mediary. The finding that postnatal androgen antagonist and insulin sensitizer treatment leads to the same outcome indicates that this resetting is mediated via increased androgen and insulin receptor signaling.[br][br]Sources of Research Support: NIH P01 HD44232 (VP).[br][br]Nothing to Disclose: AV-L, BAS, AW, CH, EB, VP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1304 174 1088 SUN-31 PO34-01 Sunday 883 2012


882 ENDO12L_SUN-32 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Increased Adult Adrenal Androgen Secretion Is Programmed by Increased Prenatal Testosterone Exposure in an Ovine Model of Polycystic Ovary Syndrome Kirsten Hogg, Fiona Connolly, Michael T Rae, Alan S McNeilly, W Colin Duncan University of Edinburgh, Edinburgh, UK; Napier Edinburgh University, Edinburgh, UK Prenatal exposure of a female fetus to increased concentrations of androgen results in an adult phenotype reminiscent of polycystic ovary syndrome (PCOS). Using an ovine model we have shown that enhanced thecal androgen production is prenatally programmed (1). The expression of enzymes involved in androgen synthesis is increased in adult theca cells and they respond to luteinizing hormone (LH) [italic]in vitro[/italic] with increased androgen secretion (1). We aimed to determine if adrenal hyperandrogenism was also developmentally programmed. Scottish Greyface ewes were injected twice weekly with 100mg testosterone propionate (TP) or vehicle control (C) from day 62 to day 102 of gestation and female offspring were studied as fetuses (day 90: TP=8, C=6), lambs (10 weeks: TP=7, C=9) or adults (11 months: TP=9, C=5). In the fetal adrenal TP increased the expression of [italic]CYP11A[/italic] (P[lt]0.05) and [italic]HSD3B1[/italic] (P[lt]0.01) but had no effect on [italic]STAR[/italic] and [italic]CYP17[/italic]. In adult offspring, fetal exposure to TP did not alter the cortisol response to intravenous synacthen (50[mu]g) but the testosterone response was increased (P[lt]0.05) after 30 mins. Fetal TP exposure increased the expression of [italic]STAR, CYP11A, HSD3B1, CYP17[/italic] (all P[lt]0.05), but not [italic]CYP21[/italic], in the adult adrenal. However in the prepubertal lamb there were no differences in the testosterone response to synacthen or in the expresssion of [italic]STAR, CYP11A, HSD3B1[/italic] or [italic]CYP17[/italic]. This confirms that the adrenal hyperandrogenism is a feature of prenatal androgenization but that it increasingly develops post-natally. This would be consistent with a possible role for hyperinsulinemia, which is more evident in adults than lambs, in this phenotype rather than a persistence of the direct effect on the fetal adrenal.[br][br](1) Hogg K et al., Endocrinology 2012; 153:45-61.[br][br]Sources of Research Support: Medical Research Council Grant G0500717; Scottish Senior Clinical Fellowship awarded to WCD.[br][br]Nothing to Disclose: KH, FC, MTR, ASM, WCD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 721 174 1089 SUN-32 PO34-01 Sunday 884 2012


883 ENDO12L_SUN-33 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Diet-Induced Inflammation Is Independent of Abdominal Adiposity in Normal Weight Women with Polycystic Ovary Syndrome (PCOS) Frank Gonzalez, Chang Ling Sia, Marguerite K Shepard, Neal S Rote, Judi Minium Indiana University School of Medicine, Indianapolis, IN; Case Western Reserve University School of Medicine, Cleveland, OH [bold]Objective: [/bold]It is unclear whether the diet-induced inflammation observed in normal weight women with PCOS is a result of increased abdominal adiposity (AA). We evaluated the status of glucose-stimulated NF[kappa]B activation and I[kappa]B protein content from mononuclear cells (MNC) of normal weight women with PCOS with and without increased AA compared to body composition-matched ovulatory controls; and their relationship with insulin sensitivity and circulating TNF[alpha], CRP and androgens.[br][bold]Materials and[/bold] [bold]Methods:[/bold] Fifteen normal weight women with PCOS (7 normal AA, 8 increase AA) between ages 18-40 years, diagnosed on the basis of oligo- or amenorrhea and hyperandrogenemia, and 16 ovulatory controls (8 normal AA, 8 increase AA) of similar age were studied. Increased AA was defined as the percent ratio of truncal fat to total body fat (% TF/TBF) measured by DEXA that was 2 SD above the mean of normal AA controls. All subjects underwent a 2 hour 75 gm oral glucose tolerance test. NF[kappa]B activation and I[kappa]B protein content were respectively quantified by electrophoretic mobility shift assay and Western blotting in MNC isolated from blood samples obtained at 0 and 2 hours. Insulin sensitivity was derived by IS[sub]OGTT[/sub].[br][bold]Results:[/bold] Compared to women with PCOS with increased AA, those with normal AA exhibited higher testosterone (88[plusmn]7 vs. 60[plusmn]5 ng/dl, p[lt]0.001) and lower TNF[alpha] (0.45[plusmn]0.08 vs. 1.73[plusmn]0.47 pg/ml, p[lt]0.003) and CRP (0.40[plusmn]0.09 vs. 1.73[plusmn]0.26 mg/l, p[lt]0.0006). Compared to body composition-matched controls, both PCOS groups exhibited lower IS[sub]OGTT[/sub] (normal AA: 4.7[plusmn]0.4 vs. 7.9[plusmn]0.9, p[lt]0.005; increased AA: 4.3[plusmn]0.6 vs. 8.3[plusmn]0.9, p[lt]0.002), greater % change in NF[kappa]B activation (normal AA: 18[plusmn]13 vs. -21[plusmn]6, p[lt]0.03; increased AA: 33[plusmn]18 vs. -17[plusmn]6, p[lt]0.005), and decreased % change in I[kappa]B protein content (normal AA: -29[plusmn]7 vs. 34[plusmn]9, p[lt]0.0001; increased AA: -20[plusmn]7 vs. 27[plusmn]10, p[lt]0.002). For the combined groups, % TF/TBF was positively correlated with TNF[alpha] (r=0.70, p[lt]0.0001) and CRP (r=0.52, p[lt]0.004). IS[sub]OGTT [/sub]was negatively correlated with % change in NF[kappa]B activation (r=-0.45, p[lt]0.02), and positively correlated with % change in I[kappa]B protein content (r=0.52, p[lt]0.003). The % change in NF[kappa]B activation was positively correlated with testosterone (r=0.43, p[lt]0.02), and androstenedione (r=0.47, p[lt]0.01).[br][bold]Conclusions:[/bold] In PCOS, the inflammatory response to glucose ingestion is independent of AA and may promote hyperandrogenism and insulin resistance in the disorder. This phenomenon is further perpetuated by increased AA.[br][br]Sources of Research Support: NIH grant HD048535.[br][br]Nothing to Disclose: FG, CLS, MKS, NSR, JM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 104 174 1090 SUN-33 PO34-01 Sunday 885 2012


884 ENDO12L_SUN-34 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Dynamics of the Pituitary Gonadal Axis in PCOS Patients during Different Stages of Reproductive Life Teresa Sir-Petermann, Amanda Ladron de Guevara, Nicolas Crisosto, Barbara Echiburu, Jessica Preissler, Manuel Maliqueo, Jose Galgani, Francisco Perez-Bravo University of Chile, Santiago, Chile; University of Chile, Santiago, Chile [bold]Context:[/bold] We have recently observed that during the 35 to 40 year old window, a period of significant decrease in fertility rates, PCOS women have a pituitary-gonadal function that is clearly different from that observed in healthy women. Nevertheless, the characteristics of the pituitary gonadal axis in younger and older PCOS patients have not been systematically assessed. [bold]Objective:[/bold] The aim of the present study was to evaluate the dynamics of the gonadal axis of PCOS patients during different stages of reproductive life. [bold]Design:[/bold] Sixty four PCOS patients who had been previously diagnosed in our laboratory according to the NIH criteria were classified into three age groups: Group A: 18 to 35 years old (n=40); Group B: 36 to 44 years old (n=15); Group C: 45 to 55 years old (n=9). Anthropometric parameters were recorded and a GnRH analog test with measurement of gonadotropins, sex steroids, SHBG and AMH levels was performed. Ovarian volumes were also determined by ultrasound[bold]. Results[/bold]: Anthropometric parameters were similar between the three groups. LH and FSH levels had a trend to rise with age, reaching significantly higher values in group C. Testosterone levels were significantly higher in group A compared to group B. AMH levels had a clear trend to decrease with age, reaching significant differences only between groups A and C (p=0.0032). Average ovarian volumes also tended to decrease with age, showing significantly lower volumes in group A compared to group B. [bold]Conclusions[/bold]: The pituitary gonadal axis of PCOS patients shows progressive changes that can be noticed along the different age groups, with rising gonadotropins and decreasing androgen and AMH levels that reflect follicle depletion. Androgen and AMH levels from group B are comparable to those observed in healthy women. Thus, this different reproductive profile may confer a window of opportunity for reproduction in these women.[br][br]Sources of Research Support: FONDECYT Grant 1110864; SOCHED (Chilean Society of Endocrinology and Diabetes)Grant 2009- 48 05.[br][br]Nothing to Disclose: TS-P, ALdG, NC, BE, JP, MM, JG, FP-B 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 90 174 1091 SUN-34 PO34-01 Sunday 886 2012


885 ENDO12L_SUN-35 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Developmental Resistance to Progesterone Negative Feedback in Hyperandrogenemic Adolescent Girls: Evidence for the Evolution of GnRH Pulse Patterns through Puberty Jennifer P Beller, Michelle Y Abshire, Christine M Burt Solorzano, Jessicah P Collins, Christopher R McCartney, John C Marshall University of Virginia Health System, Charlottesville, VA Mechanisms underlying normal puberty in girls remain unclear. Prepubertal GnRH secretion is minimal, and early puberty is marked by nocturnal increases in LH (GnRH) pulsatility. Over time, daytime LH frequency/amplitude gradually increases, leading to adult secretory patterns (1). The increase in LH secretion[mdash]in particular pulse frequency[mdash]across puberty is consistent with a progressive decline of GnRH pulse generator sensitivity to progesterone (P) negative feedback (2-4). This occurs in parallel with physiologic increases of free testosterone (T) (5), suggesting that rising T may reduce hypothalamic sensitivity to P feedback during normal puberty. Pubertal girls with hyperandrogenemia (HA) have decreased sensitivity to P negative feedback. In HA girls, elevated T levels may prematurely decrease hypothalamic sensitivity to P, resulting in increased daytime GnRH frequency and rapid 24-hour GnRH pulsatility.[br]We studied LH pulse responses to P in two HA girls across pubertal maturation. LH was measured every 10 minutes from 1900-0700h, before and after 7 days of P and estradiol administration. Hypothalamic P sensitivity was defined as the slope of % reduction of LH frequency (pulses/11 h) as a function of day 7 P level. Reference slope data (mean [plusmn] SD) for normal Tanner 1-2 = 19.3 [plusmn] 2.9; normal Tanner 3-5 = 10.2 [plusmn] 1.8; HA Tanner 3-5 = 4.69 [plusmn] 0.59.[br]The first HA subject was studied at midpuberty (Tanner 3) and had a slope of 12.7 (free T 80.3 pmol/L). One year later, when Tanner 5, she was relatively insensitive to P inhibition (slope 4.7; free T 129 pmol/L). A third assessment 18 months later showed a stable insensitivity to P inhibition (slope = 3.2; free T 84.4 pmol/L). A second subject completed two studies during late puberty (Tanner 5). Her initial slope was 6.9 (free T 27.8 pmol/L), which remained stable 18 months later at 5.3 (free T 27.3 pmol/L).[br]These results suggest that in the setting of HA, hypothalamic insensitivity to P progresses throughout puberty, remaining stable after Tanner stage 5. The data provide insight into the role of androgens in the evolution of LH/GnRH pulse patterns across puberty. The duration of exposure to HA required for development of resistance to P negative feedback remains uncertain. The results imply the presence of a time window before adult hypothalamic steroid feedback set points are established, during which preventive interventions to reduce HA in adolescent girls may be effective in restoring sensitivity.[br][br](1) McCartney CR, Prendergast KA, Blank SK, Helm KD, Chhabra S, Marshall JC 2009 Maturation of luteinizing hormone (gonadotropin-releasing hormone) secretion across puberty: evidence for altered regulation in obese peripubertal girls. J Clin Endocrinol Metab 94:56-66. (2) Blank SK, McCartney CR, Helm KD, Marshall JC 2007 Neuroendocrine effects of androgens in adult polycystic ovary syndrome and female puberty. Seminars in reproductive medicine 25:352-359. (3) Blank SK, McCartney CR, Chhabra S, Helm KD, Eagleson CA, Chang RJ, Marshall JC 2009 Modulation of GnRH pulse generator sensitivity to progesterone inhibition in hyperandrogenic adolescent girls - Implications for regulation of pubertal maturation. J Clin Endocrinol Metab 94:2360-2366. (4) Chhabra S, McCartney CR, Yoo RY, Eagleson CA, Chang RJ, Marshall JC 2005 Progesterone inhibition of the hypothalamic gonadotropin-releasing hormone pulse generator: evidence for varied effects in hyperandrogenemic adolescent girls. J Clin Endocrinol Metab 90:2810-2815. (5) McCartney CR, Blank SK, Prendergast KA, Chhabra S, Eagleson CA, Helm KD, Yoo R, Chang RJ, Foster CM, Caprio S, Marshall JC 2007 Obesity and sex steroid changes across puberty: evidence for marked hyperandrogenemia in pre- and early pubertal obese girls. J Clin Endocrinol Metab 92:430-436.[br][br]Sources of Research Support: NIH: (1) Training grant, T32 DK007646, Training in Neuroendocrinology; (2) SCCPIR grant, U54 HD28934, Clinical and Basic Studies in Polycystic Ovarian Syndrome.[br][br]Nothing to Disclose: JPB, MYA, CMBS, JPC, CRM, JCM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 769 174 1092 SUN-35 PO34-01 Sunday 887 2012


886 ENDO12L_SUN-36 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) High-Dose Vitamin D Fails To Improve Insulin Sensitivity in Women with Polycystic Ovary Syndrome Nazia Raja-Khan, Christy Stetter, Mary Lott, Allen R Kunselman, William C Dodson, Richard S Legro Pennsylvania State University College of Medicine, Hershey, PA Vitamin D deficiency may play an important role in the pathogenesis of Polycystic Ovary Syndrome (PCOS) by promoting insulin resistance and inflammation, leading to serious long-term complications including type 2 diabetes and cardiovascular disease. Our objective was to determine the effects of vitamin D on insulin resistance and inflammation in women with PCOS. We randomized 28 PCOS women in a double-blind fashion to vitamin D[sub]3[/sub] 12,000 International Units (IU) or placebo once daily for 12 weeks. Randomization was stratified for oral contraceptive (OCP) use and there were 5 OCP users in each group. None of the women were on insulin sensitizing drugs such as metformin. Before and after treatment fasting blood work, a 75 gram oral glucose tolerance test (OGTT) with blood draws at 0 and 120 minutes, and questionnaires were administered. Twenty two women completed the study. There were eleven completers per group. Results were analyzed by intention-to-treat and adjusted for OCP use and season. The two groups were similar in age and BMI (mean [plusmn] SD in vitamin D group 28.2 [plusmn] 5.2 years and 37.2 [plusmn] 4.5 kg/m[sup]2[/sup]; placebo 28.7 [plusmn] 5.6 years and 35.1 [plusmn] 9.8 kg/m[sup]2[/sup]). Compared to placebo, vitamin D significantly increased 25-hydroxy vitamin D levels (mean (95% CI) in vitamin D group 20.1 (15.7 to 24.5) ng/ml at baseline and 65.7 (52.3 to 79.2) ng/ml at 12 weeks; placebo 22.5 (18.1 to 26.8) ng/ml at baseline and 23.8 (10.4 to 37.2) ng/ml at 12 weeks; vitamin D vs. placebo, P [lt] 0.001). In the vitamin D group, there were significant reductions in the intact parathyroid hormone level (from 39.7 (26.9 to 52.4) pg/ml at baseline to 15.4 (6.5 to 24.2) pg/ml at 12 weeks, P = 0.004), but this difference did not meet statistical significance in the between groups comparison. We found no significant differences between groups in fasting insulin or glucose, 2-hour insulin or glucose, quantitative insulin sensitivity check index (QUICKI), homeostatic model assessment of insulin resistance (HOMA-IR), insulin sensitivity index (using fasting and 2-hour insulin and glucose), high sensitive C-reactive protein, free or total testosterone, lipid profiles, Positive and Negative Affect Schedule scores, and Beck Depression Inventory-II scores. In conclusion, high-dose vitamin D (12,000 IU per day) significantly increases 25-hydroxy vitamin D levels but fails to improve insulin sensitivity or inflammation in women with PCOS.[br][br]Sources of Research Support: Pennsylvania State College of Medicine Dean[apos]s Feasibility Grant and BTR Group, Inc.[br][br]Nothing to Disclose: NR-K, CS, ML, ARK, WCD, RSL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1179 174 1093 SUN-36 PO34-01 Sunday 888 2012


887 ENDO12L_SUN-37 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Androgen Receptor Antagonists Improve, and Drospirenone-Containing Oral Contraceptives Worsen, Glucose Tolerance in Women with PCOS Rodis Paparodis, Andrea E Dunaif University of Wisconsin School of Medicine and Public Health, Madison, WI; Feinberg School of Medicine, Northwestern University, Chicago, IL Polycystic ovary syndrome (PCOS) is associated with marked defects in insulin action and secretion resulting in prevalence rates of glucose intolerance as high as 40%. Previous studies have suggested that androgen receptor blockade improves insulin sensitivity in PCOS. However, the impact of oral contraceptives (OCP) on insulin sensitivity in PCOS is unclear. This study examined the effects of these agents on glucose homeostasis and body composition in women with PCOS.[br]Obese women with PCOS and normal (NGT) or impaired glucose tolerance (IGT) were enrolled in a single-blind, randomized study of flutamide (Flu) 250 mg twice daily (n=7), ethinyl estradiol (EE) 30[micro]g and drospirenone 3 mg (Yasmin[reg]) OCP (n=10), 21 d on, 7 d off, or placebo (Plc, n=12) for 6 mon. All subjects received a weight-reducing diet resulting in a deficit of 3,500 kCal/week. Oral glucose tolerance (OGTT) and frequently sampled intravenous glucose tolerance (FSIGT) testing were performed at baseline and 6 mon. Body composition was assessed by DEXA and visceral adipose tissue (VAT) by MRI. Baseline-6 mon data were compared by 1-way ANOVA or Kruskal Wallis test. Glucose tolerance was categorized as improved (IGT to NGT), unchanged or worsened (NGT to IGT) and compared by Fisher[apos]s Exact Test.[br]Body weight and fat mass decreased within the groups but only Plc was significant for fat mass (-3.2 kg, P=0.03). These parameters did not differ between groups. VAT did not differ within or between groups. At baseline, the percent NGT and IGT was: Flu 57%, 43%; OCP 30%, 70%; Plc 25%, 75%. At 6 mon, the percent with improved, unchanged or worsened glucose tolerance was: Flu 43%, 57%, 0%; OCP 0%, 60%, 40%; Plc 0%, 83%, 17%, [chi][sup2]=15.24, P=0.0042. FSIGT-measures of insulin sensitivity and secretion did not change within or between groups.[br]The administration of an OCP with relatively low dose EE and an antiandrogenic progestin, drospirenone, worsened glucose tolerance in obese women with PCOS. In contrast, androgen receptor blockade improved it. There were no significant changes in insulin sensitivity or secretion, so the effects on glucose tolerance may have been mediated by alterations in glucose clearance. These findings suggest that caution should be exercised when administering OCPs to obese women with PCOS, particularly women with glucose intolerance. In contrast, the potential benefits of androgen receptor blockade on glucose intolerance in PCOS merit further investigation.[br][br]Sources of Research Support: NIDDK.[br][br]Nothing to Disclose: RP, AED 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1333 174 1094 SUN-37 PO34-01 Sunday 889 2012


888 ENDO12L_SUN-38 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Impaired Positive Feedback Actions of Progesterone in PCOS: A Potential Contributor to Oligo-/Anovulation Michelle Y Abshire, Jennifer P Beller, Jessicah SP Collins, Christopher R McCartney University of Virginia Health System, Charlottesville, VA; University of Virginia Health System, Charlottesville, VA Progesterone (P) is the primary mediator of GnRH pulse frequency slowing during the luteal phase[mdash]a reflection of P-negative feedback (1). P-[italic]positive[/italic] feedback appears to be important for full expression of midcycle LH release. For example, P increases shortly before the LH surge, and P augments LH release in the presence of late follicular estradiol (E2) levels (2). P also acutely increases LH pulse amplitude and mean LH in E2-pretreated women during the late follicular phase (3).[br]Women with polycystic ovary syndrome (PCOS) display reduced GnRH pulse generator sensitivity to P-negative feedback. We hypothesized that PCOS would also be marked by reduced sensitivity to P-positive feedback. To test this, we assessed P-induced changes of LH in E2-pretreated women with PCOS (n=3) and normally cycling controls (n=7).[br]Women underwent two frequent sampling admissions, each lasting from 2000-1400h. E2 (transdermal patch, 0.2 mg/d) was given 3 days before and throughout each admission. Control women were admitted on cycle days 7-11; women with PCOS were at least cycle day 7 at admission (luteal phase excluded by serum P [lt] 1 ng/ml). At 0600 h during admission, women received either oral micronized P (100 mg) or placebo (determined by randomization). At least 26 days thereafter, women underwent a second sampling study identical to the first, except that women initially receiving P received placebo and [italic]vice versa[/italic]. We analyzed mean LH and LH amplitude before and after P/placebo (i.e., 2200-0600 h vs. 0600-1400 h). To account for possible diurnal changes, apparent changes after P were normalized to apparent changes after placebo.[br]Compared to controls, women with PCOS had 3-fold higher mean free T and 45% higher mean BMI. Achieved E2 levels were nearly 2-fold higher in PCOS, but LH amplitude and mean LH from 2200-0600 h were similar between PCOS and controls. Despite higher E2 in PCOS, the fold increase of LH pulse amplitude attributable to P was only 2.32 [plusmn] 1.51 in PCOS, less than half of that in controls (4.83 [plusmn] 1.76). Similarly, the fold increase of mean LH attributable to P was 1.42 [plusmn] 0.28 in PCOS vs. 2.56 [plusmn] 0.43 in controls. These data suggest that acute P administration provokes a robust increase of LH pulse amplitude and mean LH in normal E2-pretreated women, but that this effect is blunted in women with PCOS. This may reflect relative resistance to the positive feedback effects of P, which could represent another factor contributing to oligo-/anovulation in PCOS.[br][br](1) Soules MR et al., J Clin Endocrinol Metab 1984; 58: 378-383. (2) Chang, RJ and Jaffe RB., J Clin Endocrinol Metab 1978; 47: 119-125. (3) McCartney, CR et al., Am J Physiol Endocrinol Metab 2007; 292: E900-906.[br][br]Sources of Research Support: NIH R01 HD058671.[br][br]Nothing to Disclose: MYA, JPB, JSPC, CRM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 207 174 1095 SUN-38 PO34-01 Sunday 890 2012


889 ENDO12L_SUN-39 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Impaired Autonomic Response in Patients with Different Phenotypes of Polycystic Ovary Syndrome Kristhiane Di Domenico Cunha, Denusa Wiltgen, Fabian Nickel, Ruy Silveira Moraes, Poli Mara Spritzer Hospital de Clinicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil; Hospital de Clinicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil; Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil Women with Polycystic Ovary Syndrome (PCOS), a prevalent endocrine disease characterized by ovulatory dysfunction and hyperandrogenism, have a higher risk for development of metabolic, cardiovascular and autonomic abnormalities. Heart rate variability (HRV) is a useful method to evaluate autonomic modulation integrity at rest and the response to sympathetic stimulation. Therefore, the aim of this study was to evaluate autonomic modulation at rest and during sympathetic stimulation in patients with different PCOS phenotypes and healthy controls. In this cross-sectional study, 30 classic PCOS (c-PCOS) [ovary dysfunction and hyperandrogenism], 16 ovulatory PCOS (ov-PCOS) [normal androgens, hirsutism and with polycystic ovaries (PCO)] and a control group (C) of 23 nonhyperandrogenic ovulatory women were studied. Vagal [mean RR interval (MRR), root mean square of successive differences (RMSSD), percentage of differences between RR intervals exceeding 50 ms (pNN50), high frequency component (HF)] and sympathetic (low frequency component (LF) and LF/HF ratio) HRV indices were calculated using a 30 minute Holter recording [20 minutes of supine rest followed by 10 minutes of mental stress (Stroop Color-word test)]. All subjects were similar in age and had similar physical activity status assessed by a pedometer. c-PCOS patients had higher BMI (c-PCOS 31.47 [plusmn] 5.21; ov-PCOS 24.47 [plusmn] 3.62; C 27,18 [plusmn] 5,36; p[lt]0.01) and total testosterone levels (c-PCOS 0.81 [plusmn] 0.25; ov-PCOS 0.62 [plusmn] 0.20; C 0.54 [plusmn] 0.17; p[lt]0.01), compared with ov-PCOS and controls. HRV was similar between groups, during rest. Mental stress reduced MRR, RMSSD, pNN50 and HF, and increased LF and LF/HF ratio in all sample but, PCOS patients, after adjustment for BMI and age, responded with less vagal withdrawal [MRR (cPCOS 669 (637-761); ov-PCOS 652,55 (615-734); C 629 (605-665); p=0.021), PNN50 (cPCOS 4 (2-16); ov-PCOS 3.25 (0.25-10); C 1.82 (0.53-8); p=0.016), and RMSSD (cPCOS 24 (20-40); ov-PCOS 24 (16-31); C 20 (16-29); p=0.016)] and sympathetic predominance than controls presenting with lower LF than C group (cPCOS 0.70 (0.61-0.81); ov-PCOS 0.72 (0.61-0.88); C 0.88 (0.81-1.52); p=0.00), regardless of phenotype. In conclusion, PCOS patients have impaired autonomic response to sympathetic stimulation, especially in the most complete phenotype.[br][br]Sources of Research Support: Brazilian National Institute of Hormones and Women[apos]s Health.[br][br]Nothing to Disclose: KDC, DW, FN, RSM, PMS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 266 174 1096 SUN-39 PO34-01 Sunday 891 2012


890 ENDO12L_SUN-40 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Trp28Arg/Ile35Thr LHB Gene Variants Are Associated with Higher Testosterone Levels in Brazilian Women with Polycystic Ovary Syndrome Mariani Carla Prudente Batista, Eliane de Fatima Duarte, Michele Delarmelina dos Reis Borba, Emilie Zingler, Carla Capanema, Beatriz Taynara Araujo dos Santos, Olivia Laquis, Sylvia Hayashida, Francisco de Assis Rocha Neves, Edmund C Baracat, Gustavo AR Maciel, Tania ASS Bachega, Gustavo B Barra, Adriana Lofrano-Porto University Hospital of Bras[iacute]lia (HUB/UnB), Bras[iacute]lia, Brazil; University of Bras[iacute]lia, Bras[iacute]lia, Brazil; University of Bras[iacute]lia (UnB), Bras[iacute]lia, Brazil; Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Sabin Laboratory and Institute, Bras[iacute]lia, Brazil INTRODUCTION: Polycystic ovary syndrome (PCOS) is a complex metabolic disease, which affects 6-10% of women of reproductive age leading to menstrual abnormalities, infertility and hirsutism. The genetic basis of PCOS is still unknown despite significant family clustering, and the wide variability in clinical presentation makes it difficult to determine the mode of inheritance. Although recent studies have failed to reveal a causal link between LHB gene and development of PCOS, its crucial functional roles in reproductive disorders still makes it a natural candidate. In this study, we analyzed the association of LHB gene polymorphisms with hyperandrogenic phenotype in PCOS. METHODS: 130 Brazilian PCOS women, aged 14 to 42 years, meeting the 2004 Rotterdam criteria were included. Hirsutism was defined by Ferriman-Gallwey Score (FGS) [ge]8. Body mass index, fasting insulin and glucose, LH, FSH and total testosterone (TT) levels were assessed. Subjects were divided according to the presence and severity of hirsutism (absent [FGS[lt]8], mild [FGS 8-15] and severe [FGS[gt]15], as well as to TT levels (normal or high, i.e., above the upper normal limit for the method). The coding region of LHB gene was sequenced in all patients and also in 96 normal women. Chi-square and Fisher[apos]s exact test were used to test the association between LHB variants and PCOS phenotype, deemed statistically significant at the level of p[lt]0.05. RESULTS: Eleven polymorphisms were detected in the LHB coding region. Their frequencies were in Hardy-Weinberg equilibrium and did not differ between patients and controls. The Trp28Arg/Ile35Thr variants are in linkage disequilibrium and were found in 87.1% and 86.4% of the patients and controls alleles, respectively. Frequency of LHB variants did not differ according to metabolic profile, presence of hirsutism and its severity. Interestingly, in women presenting high TT levels, the prevalence of Trp28Arg/Ile35Thr polymorphism was significantly higher than the wild type genotype (p=0.015). CONCLUSIONS: These results are in agreement with in vitro studies showing a higher bioactivity of these LHB variants, despite a shorter half life. Although a causal role for LHB polymorphisms in PCOS has not been determined, our results suggest a modulating effect of LHB variants in the hyperandrogenemia of SOP, which could possibly explain some of the variability in the clinical presentation of the syndrome.[br][br]Sources of Research Support: Funda[ccedil][atilde]o de Apoio [agrave] Pesquisa do Distrito Federal (FAPDF), n[uacute]mero do projeto 323/2011, 193000407/2010.[br][br]Nothing to Disclose: MCPB, EdFD, MDdRB, EZ, CC, BTAdS, OL, SH, FdARN, ECB, GARM, TASSB, GBB, AL-P 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2100 174 1097 SUN-40 PO34-01 Sunday 892 2012


891 ENDO12L_SUN-41 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Prevalence of Polycystic Ovary Syndrome in North Indians [mdash] A Community Survey Preeti Dabadghao, Harmandeep Gill, Pallavi Tiwari Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, India Background: Polycystic Ovary Syndrome (PCOS) is considered one of the most common endocrine disorders in reproductive age group. Due to the logistics of diagnosis and lack of consensus on the diagnostic criteria, there are very few prevalence studies in the community. This study was aimed to assess the prevalence of PCOS in women 18-25 years of age, conducted in college girls from Lucknow, north India.[br]Methods: Sample size for the study was calculated as 1052. Girls from 3 different colleges were approached (n=2150), 1520 (71 %) agreed to participate. They were asked to fill up a questionnaire asking details of menstrual cycle and features of hyperandrogenism. Hirsutism was self-reported. Responses were verified by a trained research assistant. A probable case was defined as a girl with menstrual irregularity (MI) or hirsutism (H) or both. All the probable cases were invited for detailed examination, hormone estimation and ovarian ultrasonography.[br]Results: Of the 1520 girls, 200 (13.1%) were labeled as probable cases; 175 (87.5%) had MI and 25 (12.5%) had both MI and H. The girls tended to over- report hirsutism. Of the 25 who reported to have hrsutism, only 3of 13 examined had clinically significant hirsutism, modified Ferriman Gallway score of [gt]6. Of these 200 cases, 74(37%) girls have had hormonal evaluation while 12 agreed for ultrasonography. Twenty five girls have been confirmed to have PCOS according to either National Institute of Health or Rotterdam criteria. So if all the 200 girls would have had hormone evaluation and/or ultrasonography, 68 girls were likely to be confirmed as PCOS, giving a calculated prevalence of 4.4% in this population.[br]The mean age of the PCOS cases was 19.0[plusmn]1.3 yrs, body mass index was 21.2[plusmn]5.2 Kg/m[sup]2[/sup] and waist hip ratio was 0.79[plusmn] 0.06. Only 16% girls had a body mass index [gt]23 Kg/m[sup]2[/sup], but 40% had waist hip ratio [gt]0.81, again highlighting that despite low BMI, Indians have more abdominal obesity. None of the girls was hypertensive but 20% had prehypertension. There was no difference in any of the clinical or biochemical parameters (except androgen levels) between PCOS girls, probable PCOS and 90 controls (taken from the same population) studied.[br]Conclusion: Calculated prevalence of PCOS in women between the ages of 18-25 years from Lucknow, north India is 4.4%. Majority of these girls were lean but have abdominal obesity.[br][br]Sources of Research Support: Funded by an extramural grant from UP council of Science and Technology, Lucknow, India.[br][br]Nothing to Disclose: PD, HG, PT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1045 174 1098 SUN-41 PO34-01 Sunday 893 2012


892 ENDO12L_SUN-42 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Impact of Dietary Advanced Glycation End Products (AGEs) Modifications on Metabolic and Hormonal Profile in Women with Polycystic Ovary Syndrome Evangelia Tantalaki, Christina Piperi, Sarantis Livadas, Anastasios Kollias, Christos Adamopoulos, Aikaterini Koulouri, Charikleia Christakou, Evanthia Diamanti-Kandarakis Medical School, National and Kapodistrian University of Athens, Athens, Greece; Medical School, National and Kapodistrian University of Athens, Athens, Greece OBJECTIVE: To investigate the impact of dietary intervention of Advanced Glycation End products (AGEs) intake on hormonal and metabolic profile in women with polycystic ovary syndrome (PCOS).[br]METHODS: After baseline evaluation, 23 women with PCOS [mean[plusmn]SD, age: 23.4[plusmn]5.7 years; body mass index (BMI): 26[plusmn]5.7 kg/m2] underwent the following consecutive 2-month dietary regimens: a hypocaloric diet with ad-libitum AGEs content (Hypo), an isocaloric diet with high AGEs (HA) and an isocaloric diet with low AGEs (LA). Metabolic, hormonal and oxidative stress status was assessed as well as AGEs levels were determined in all subjects, after the completion of any dietary intervention.[br]RESULTS: Serum levels of AGEs, testosterone, oxidative stress, insulin and HOMA-IR index were significantly increased on HA compared to Hypo diet and subsequently decreased on LA diet (compared to HA) (p[lt]0.05 for all parameters). BMI remained unaltered throughout the HA and LA periods compared to Hypo period. Dietary AGEs were associated with serum AGEs (r=0.45, p[lt]0.001), insulin (r=0.3, p=0.005), HOMA-IR index (r=0.31, p=0.006) and oxidative stress (r=0.32, p=0.002). Testosterone was correlated with insulin (r=0.36, p=0.001) and HOMA-IR index (r=0.36, p=0.002).[br]CONCLUSIONS: Modifications on dietary AGEs intake are associated with parallel changes in serum AGEs, metabolic, hormonal and oxidative stress biomarkers in women with PCOS. These findings support the advice of a low AGEs dietary content along with lifestyle changes in women with PCOS.[br][br]Nothing to Disclose: ET, CP, SL, AK, CA, AK, CC, ED-K 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1406 174 1099 SUN-42 PO34-01 Sunday 894 2012


893 ENDO12L_SUN-43 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Ethinyl Estradiol Cyproterone Acetate Versus Low-Dose Pioglitazone-Flutamide-Metformin for Adolescent Girls with Androgen Excess: Divergent Effects over 12 Months Lourdes Ibanez, Marta Diaz, Matilde Rodriguez-Chacon, Elsa Maymo-Masip, Cristina Salvador, Abel Lopez-Bermejo, Joan Vendrell, Francis E de Zegher University of Barcelona [amp] CIBERDEM, ISCIII, Esplugues, Spain; Hospital Dr Josep Trueta [amp] Girona Institute for Biomedical Research, Girona, Spain; University Hospital of Tarragona Joan XXIII, Pere Virgili Institute [amp] CIBERDEM, ISCIII, Tarragona, Spain; University of Leuven, Leuven, Belgium; University of Barcelona, Esplugues, Spain [bold]Objective [/bold][br]To compare the effects of a traditional therapy to those of a novel treatment for androgen excess in adolescent girls.[br][bold]Study Design [/bold][br]Randomized, open-label trial over 12 months.[br][bold]Study Participants[/bold][br]Non-obese adolescents (mean age 16 yr, BMI 23 Kg/m[sup]2[/sup]) with hyperinsulinemic androgen excess and without risk of pregnancy.[br][bold]Interventions[/bold][br]EthinylEstradiol-CyproteroneAcetate (EE-CA; Diane 35 Diario) versus a low-dose combination of pioglitazone 7.5 mg/d, flutamide 62.5 mg/d, and metformin 850 mg/d (PioFluMet).[br][bold]Main Outcome Measures[/bold][br]Hirsutism and acne scores; androgen excess; circulating C-reactive protein and high-molecular-weight adiponectin; carotid intima-media thickness; body composition (by absorptiometry); abdominal fat partitioning (by MRI); gene expression in biopsies of subcutaneous adipose tissue at the abdominal level.[br][bold]Results[/bold][br]EE-CA and PioFluMet reduced the clinical and biochemical androgen excess comparably but had divergent effects on C-reactive protein and high-molecular-weight adiponectin; on carotid intima-media thickness; on lean mass; on abdominal and visceral fat; and on the expression of genes such as CD163, TNF-like weak inducer of apoptosis (TWEAK) and ANGPTL4 that relate to macrophage activation, inflammation and lipoprotein metabolism. All these divergences pointed to a healthier condition on low-dose PioFluMet.[br][bold]Conclusion[/bold][br]Over 12 months, the effects of low-dose PioFluMet compared favorably to those of EE-CA in adolescents with androgen excess.[br][br]Sources of Research Support: A grant (#PI09/90444) from the Ministerio de Ciencia e Innovacion, Instituto de Salud Carlos III, Madrid, Spain.[br][br]Nothing to Disclose: LI, MD, MR-C, EM-M, CS, AL-B, JV, FEdZ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 310 174 1100 SUN-43 PO34-01 Sunday 895 2012


894 ENDO12L_SUN-44 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Visceral Adiposity Index as an Indicator of Insulin Resistance in Women with Polycystic Ovary Syndrome Jee-Young Oh, Yeon-Ah Sung, Hye Jin Lee Ewha Womans University School of Medicine, Seoul, Korea Although insulin resistance is a major pathophysiology of polycystic ovary syndrome (PCOS), its assessment is clinically limited. Recently, visceral adiposity index (VAI) is introduced as a surrogate marker of visceral adipose dysfunction. It has been reported to be associated with the rate of peripheral glucose utilization during euglycemic hyperinsulinemic clamp. We aimed to evaluate the VAI could be useful marker for insulin resistance in Korean women with PCOS.[br]We recruited 736 women with PCOS diagnosed by 1990 NIH criteria. Anthropometric measurements and metabolic and hormonal parameters were obtained. Glucose tolerance status was assessed by 75 g OGTT. Insulin sensitivity index was used by metabolic clearance rate (MCR) of glucose during OGTT. VAI was calculated as [(waist/36.58+(1.88xBMI)] x (TG/0.81) x (1.52/HDL).[br]The Mean age and BMI were 23[plusmn]5 (15[sim]39 yrs) and 22.8[plusmn]4.3 (15.5[sim]39.4 kg/m[sup]2[/sup]). VAI was significantly correlated with fasting insulin (r=0.25), 2h-postload insulin (r=0.34), AUC insulin (r=0.25), HOMA-IR (r=0.25), and MCR (r=-0.55), and it also significantly correlated with free testosterone (r=0.38) and free androgen index (r=0.41, all Ps[apos] [lt]0.0001). When we defined the insulin resistance as [lt] 5 percentile of MCR in regular cycling control women, VAI was significantly associated with insulin resistance after adjustment of age, systolic blood pressure, 2h-postload glucose, and total cholesterol (OR 1.4, 95% CI 1.1[sim]1.7).[br]In conclusion, VAI could be helpful to predict the insulin resistance in young Korean women with PCOS.[br][br]Nothing to Disclose: J-YO, Y-AS, HJL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 513 174 1101 SUN-44 PO34-01 Sunday 896 2012


895 ENDO12L_SUN-45 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) The Relationship between Insulin-Like Growth Factor-I and Insulin Resistance in Patients with Hirsutism Evrim Cakir, Oya Topaloglu, Nujen Colak, Basak Karbek, Askin Gungunes, Muyesser Sayki, Ilknur Unsal, Esra Tutal, Bekir Ucan, Tuncay Delibasi Diskapi Training and Research Hospital, Ankara, Turkey Hyperinsulinemia is known frequent finding in patients with hirsutism especially in polycystic ovary syndrome (PCOS), and this factor has cause-consequence relationships with increased cardiovascular disease risk. It is not known the role of insulin like growth factor I (IGF-I) level in hepatic insulin resistance in patients with hirsutism, since the liver is the main site of its production. Therefore, IGF-I levels, fasting glucose, fasting insulin, liver function tests and antropometric measurement were evaluated in patients with idiopathic hirsutism (IH) (n=33), in PCOS (n=25) and in healthy controls (n=25). Insulin resistance was calculated by using the homeostasis model assessment insulin resistance index (HOMA-IR, according to the formula, fasting plasma glucose (mmol/L) x fasting serum insulin (mU/mL)/22.5 (1). Mean age, body mass index (BMI) and waist hip ratio were similar between groups (p[gt]0.05). Mean IGF-I levels and median ALT levels were higher in patients with IH and PCOS than controls but these differences were not statistically significant (p=0.12, p=0.26, respectively). IGF-I levels were found negatively correlated with age (r:-0.471, p[lt]0.05) whereas positively correlated with ferrimann gallwey (FG) hirsutism score (r:0.232, p[lt]0.05). After adjusted for age and body mass index (BMI), IGF-I levels were significantly higher in patients with IH and PCOS patients (p[lt]0.05). ALT level was found positively correlated with BMI (r:0.466, p[lt]0.05), FG (r:0.248, p[lt]0.05), insulin (r:0.469, p[lt]0.05), HOMA-IR (r:0.394, p[lt]0.05) and total testosterone (r:0.280, p[lt]0.05). These data show that IGF-I and ALT levels are increased in subjects with IH and PCOS. This finding suggests that increased hepatic production of IGF-I and ALT might be an early indicator of insulin resistance in hirsutism.[br][br]1.Matthews DR et al. Diabetologia 1985;28:412-419.[br][br]Nothing to Disclose: EC, OT, NC, BK, AG, MS, IU, ET, BU, TD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 616 174 1102 SUN-45 PO34-01 Sunday 897 2012


896 ENDO12L_SUN-46 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Diagnosis, Screening for Comorbidities, and Management of Polycystic Ovarian Syndrome (PCOS) across Three Disciplines within an Academic Children[apos]s Hospital Bethany A Auble, Andrea Gross, Deborah Elder, Jennifer B Hillman Cincinnati Children[apos]s Hospital, University of Cincinnati, Cincinnati, OH; Cincinnati Children[apos]s Hospital, University of Cincinnati, Cincinnati, OH CONTEXT: Polycystic ovarian syndrome (PCOS) is a complex condition associated with metabolic and reproductive health concerns. Determining whether differences exist across disciplines in the management of adolescents with PCOS is necessary to assess the standardization of care.[br]OBJECTIVE: The objective was to evaluate differences across disciplines regarding diagnosis, screening for comorbidities, and management of PCOS among adolescents.[br]METHODS: This retrospective medical record review of patients with PCOS drew from three disciplines that manage PCOS within an academic children[apos]s hospital: pediatric endocrinology (PEndo), pediatric and adolescent gynecology (PGyn), and adolescent medicine (AMed). Patients were identified by a query of billing data between July 1, 2008, and June 30, 2010. Patients identified from PEndo totaled 216, with 168 from PGyn and 97 from AMed. Power analyses estimated 57 charts from each discipline were needed to detect a difference (B=.20, p[lt].05). Charts were randomly selected for review. An abstraction tool was used to collect demographics, symptoms, anthropometrics, laboratory assessment, and treatment recommendations. Chi-square testing was used and Bonferroni correction was applied to account for multiple comparisons.[br]RESULTS: No significant differences were noted across disciplines in the use of established diagnostic criteria for PCOS. Notably, 20% of patients seen by PGyn and AMed and 13% of PEndo patients did not meet Rotterdam nor NIH Consensus criteria for PCOS. There were significant comorbidity differences across disciplines for type 2 diabetes screening (p=.006), obesity (p=.01), and insulin resistance (p=.002). Obesity was screened in 47% PEndo, 40% PGyn, and 22% AMed patients. Significant treatment differences across disciplines were noted for lifestyle changes (p=[lt].0001), oral contraceptives (p=0.015), metformin (p=.0001), anti-androgens (p=.004), and dietician referral (p=.013). PEndo recommended lifestyle changes most often. Oral contraceptives were used more frequently by PGyn (97%) and AMed (95%), than by PEndo (83%). Metformin use was highest in PEndo (90%) compared to PGyn (38%) and AMed (25%).[br]CONCLUSION: Differences exist across disciplines that commonly manage PCOS, likely reflecting training and comfort level with particular aspects of the disorder. A multidisciplinary clinical model would standardize care, impact comorbidity screening, and delineate long-term outcomes for adolescents with PCOS.[br][br]Nothing to Disclose: BAA, AG, DE, JBH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 64 174 1103 SUN-46 PO34-01 Sunday 898 2012


897 ENDO12L_SUN-47 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Relationship between Smoking and Parameters of Metabolic Syndrome in Women with Polycystic Ovary Syndrome and Controls Cindy Ta Pau, Candace Cowperthwaite Keefe, Corrine Kolka Welt Massachusetts General Hospital - Harvard Medical School, Boston, MA BACKGROUND: Smoking has been shown to decrease androgen and estrogen levels in animal models, but the effects of smoking on human reproductive hormones in premenopausal women have been conflicting. Furthermore, the relationship between smoking and metabolic syndrome parameters in women with polycystic ovary syndrome (PCOS) has not been examined.[br]METHODS: Women with PCOS, diagnosed according to the NIH criteria, and healthy controls, aged 18-40 years, were recruited and categorized according to smoking status: PCOS smokers (n=47) and non-smokers (n=72), and control smokers (n=49) and non-smokers (n=14). Physical examination, history, anthropomorphic measurements, and pelvic ultrasound were performed. Serum cotinine, nicotine, sex steroids, gonadotropins, and metabolic parameters were measured. Comparisons were made using two-way ANOVA, and relationships with cotinine and nicotine were examined using Spearman correlations.[br]RESULTS: Smokers had higher cotinine (160[plusmn]117 vs. 2.4[plusmn]1.7 ng/mL; p[lt]0.001) and nicotine levels (7.3[plusmn]6.2 vs. 2.0[plusmn]0.2 ng/mL; p[lt]0.001) than non-smokers. There were no differences in sex steroids between smokers and non-smokers. Women with PCOS had higher waist circumference (99.2[plusmn]1.8 vs. 84.7[plusmn]2.8 cm; p[lt]0.001), fasting glucose (100.0[plusmn]3.0 vs. 87.7[plusmn]4.4 mg/dL; p=0.02) and HOMA-IR (2.7[plusmn]0.3 vs. 1.4[plusmn]0.5; p=0.02), and lower HDL (52.4[plusmn]1.6 vs. 63.1[plusmn]2.2 mg/dL; p[lt]0.001) than controls, but there was no effect of smoking on these parameters. Triglycerides were higher in women with PCOS compared to controls (112[plusmn]6.vs. 70.6[plusmn]8.8 mg/dL; p[lt]0.001), and smokers had higher triglyceride levels than non-smokers (104[plusmn]72 vs. 82.8[plusmn]7.8 mg/dL; p=0.003). Smokers also had higher systolic (114.3[plusmn]1.5 vs. 108.8[plusmn]2.1 mm Hg; p[lt]0.01) and diastolic blood pressures (71.6[plusmn]10.7 vs. 67.8[plusmn]11.5 mm Hg; p[lt]0.05). There was no interaction between PCOS status and smoking with triglyceride levels, systolic blood pressure, or diastolic blood pressure. There was a correlation between cotinine (r=0.3; p=0.002) and nicotine levels (r=0.2; p[lt]0.01) with triglycerides.[br]CONCLUSION: Smokers had higher triglycerides, systolic and diastolic blood pressures. In addition, women with PCOS had higher triglycerides, waist circumference, fasting glucose, and HOMA-IR, and lower HDL compared to controls. The worsening of metabolic parameters does not appear to be related to differences in androgen levels. In conclusion, smoking worsens the already high risk for metabolic syndrome in women with PCOS.[br][br]Sources of Research Support: Flight Attendant Medical Research Institute; NIH/NICHD 1R01HD065029; American Diabetes Association 1-10-CT-57; Harvard Clinical and Translational Science Center 1 UL1 RR025758.[br][br]Nothing to Disclose: CTP, CCK, CKW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1779 174 1104 SUN-47 PO34-01 Sunday 899 2012


898 ENDO12L_SUN-48 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Non-Invasive Prenatal Diagnosis of Congenital Adrenal Hyperplasia Maria Iandolo New, Oksana Lekarev, Tony Yuen, Dennis Lo Mount Sinai School of Medicine, New York, NY; Chinese University of Hong Kong, Hong Kong, China Congenital Adrenal Hyperplasia (CAH) owing to steroid 21 hydroxlase deficiency results in genital ambiguity in females with the classical form of this autosomal recessive monogenic disorder. Genital ambiguity, which is the result of excessive fetal androgen production, is preventable by prenatal low-dose dexamethasone administered to the mother. Prenatal treatment with dexamethasone must begin before the 9[sup]th[/sup] week of gestation to prevent genital ambiguity, yet chorionic villus sampling and amniocentesis to obtain fetal DNA cannot be done until the 10[sup]th[/sup] and 12[sup]th[/sup] week of gestation respectively. While all males both affected and unaffected and unaffected females do not require prenatal treatment, currently utilized invasive methods require that all fetuses at risk for CAH must be treated for several weeks before their sex and/or affection status are known. This results in unnecessary treatment of males and unaffected females for several weeks. The discovery of circulating fetal DNA in maternal plasma has opened up new possibilities for noninvasive prenatal diagnosis for CAH. Hence, prenatal diagnosis of CAH owing to 21-hydroxylase deficiency (21OHD) can be performed at 7 weeks of gestation utilizing free fetal DNA circulating in maternal plasma. We propose to utilize fetal DNA circulating in the maternal plasma to determine (1) the sex of the fetus and thus avoid unnecessary treatment of males, and (2) the affection status of the fetus, making treatment of unaffected females unnecessary.[br]Once the DNA has been harvested from mother[apos]s plasma, prenatal diagnosis of CAH is complicated by the presence of a pseudogene which is 97% of homologous to the active Cyp21A2 gene. Thus special procedures are required to distinguish the active gene from the pseudogene. Recent advances in molecular counting approaches for analyzing circulating fetal DNA, including digital PCR and massively parallel sequencing, would be explored to address this challenging task.[br][br]Sources of Research Support: Grant from Hong Kong.[br][br]Nothing to Disclose: MIN, OL, TY, DL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 983 174 1105 SUN-48 PO34-01 Sunday 900 2012


899 ENDO12L_SUN-49 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Mathematical Modeling of the Role of Androgens in Regulation of the Ovulatory Cycle of Women Angelean O Hendrix, Claude L Hughes, James F Selgrade North Carolina State University, Raleigh, NC; Duke University Medical Center, Durham, NC Based on detailed hormonal data from cycling women and established modes of hormone action, mathematical models were first developed and applied to endocrine regulation of the pituitary-ovarian axis in women by Schlosser and Selgrade in 1999. The resulting model used biochemical properties of Luteinizing Hormone (LH), Follicle Stimulating Hormone (FSH), Estradiol (E2), Progesterone (P4), and Inhibin (INH) for stable, periodic in-silico representation. With successful parameter estimation, the Schlosser/Selgrade models accurately reflected mean serum levels and oscillatory behavior as reported by McLachlan et al. (1990). Subsequent model incarnations of Harris-Clark et al. (2003) and Pasteur and Selgrade (2011) merged the pituitary and ovarian sub-systems and expanded the model to represent both Inhibin A (INH-A) and Inhibin B (INH-B), respectively. Thorough analysis of the models by Selgrade et al. (2009) revealed an additional stable periodic solution in the five-hormone model that resembles hormone levels of patients with Polycystic Ovarian Syndrome (PCOS). Because PCOS is seen as primarily a hyper-androgenic disorder, the inclusion of androgens into the Schlosser/Selgrade model is theoretically necessary to produce accurate simulations of biologically and clinically important pituitary and ovarian hormones in women with PCOS. As Testosterone (T) is the dominant potent female androgen and is significantly increased in PCOS patients, we focused our efforts on modeling pituitary feedback and intra-ovarian follicular growth properties as functions of circulating total T levels as reported by Sinha et al. (1998). Structurally based on in-vitro findings of Yasin et al. (1996) and Weiss et al. (2007), parameters have been identified that simultaneously simulate LH, FSH, E2, P4, INH-A, and INH-B levels of Welt et al. (1999) and total T levels of Sinha et al. The resulting model expands that of Selgrade et al. to a system of twenty ordinary differential equations, with at least one stable periodic solution. Preliminary in-silico experiments designed to simulate hyper-androgenism produce approximations of the seven hormones reflecting observations in PCOS patients with increased accuracy. The new model will allow clinical investigators to study potential hormone interventions to return acyclic patients to regular ovulatory cycles.[br][br]Sources of Research Support: NSF grant DMS-0920927. National Science Foundation Graduate Research Fellowship.[br][br]Nothing to Disclose: AOH, CLH, JFS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 157 174 1106 SUN-49 PO34-01 Sunday 901 2012


900 ENDO12L_SUN-50 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) The Relationship between Insulin Resistance, TSH and Thyroid Volume in Patients with PCOS Evrim Cakir, Oya Topaloglu, Nujen Colak, Basak Karbek, Askin Gungunes, Muyesser Sayki, Ilknur Unsal, Esra Tutal, Bekir Ucan, Mustafa Sahin, Tuncay Delibasi Diskapi Yildirim Beyazit Training and Research Hospital, Ankara, Turkey In recent studies presence of insulin resistance, obesity and metabolic syndrome has found as independent risk factors for enlarged thyroid volume (TV). Insulin resistance and obesity are frequent findings in patients with PCOS. Therefore, the aim of this study was to evaluate the relationship between biochemical, hormonal parameters and thyroid volume in patients with PCOS (n=47) and healthy control (n=30). Thyroid volume was assessed using a high-resolution ultrasound machine (Hitachi, Japon; EUB 7000) with a 6[ndash]14 megahertz high-frequency linear transducer. Insulin resistance was calculated by using the homeostasis model assessment insulin resistance index (HOMA-IR, according to the formula, fasting plasma glucose (mmol/L) x fasting serum insulin (mU/mL)/22.5. The cut off value was taken as 2.7 for HOMA-IR.Mean age (control; 22.8[plusmn]4.4, PCOS; 23.2[plusmn]5.8), BMI (control; 23.6[plusmn]3.35 kg/m2, PCOS; 24.4[plusmn]4.3 kg/m2), thyroid stimulan hormone (TSH) levels (control; 1.98[plusmn]1.04 [micro] IU/ml, PCOS; 2.01[plusmn]1.19 [micro] IU/ml) and TV were similar between groups (p[lt]0.05). The TV, measured by thyroid ultrasonography (US), showed the following values: PCOS group; 10.1[plusmn]3.9 mL; Control group; 10.4[plusmn]4.3 mL. Insulin resistance, calculated with HOMA-IR, and free T4 levels was found to be higher in PCOS patients (p:0.014, p: 0.007, respectively). Thyroid volume was found positively correlated with LH (r:0.316, p:0.007) and anti TPO (r:0.602, p[lt]0.0001) levels. There was no correlation between thyroid volume, fasting insulin and HOMA-IR in correlation analyses and lineer regression analyses. Participants were divided into 2 groups according to HOMA-IR value. Group 1; HOMA-IR[ge] 2.7, Group 2; HOMA-IR[lt] 2.7. Mean TSH level was found to be higher in patients with high HOMA-IR levels (Mean TSH level was 1.84[plusmn]0.99 [micro] IU/ml in group 1 and 2.75[plusmn]1.47 [micro] IU/ml in group 2, p:0.047). Although TSH levels were found to be higher in high HOMA-IR group, mean TV was similar between groups (p[gt]0.05).[br]In early age PCOS patients it was observed that insulin resistance had no effect on TV. In this case, anti TPO and LH levels have dominant effect on TV.[br][br]Nothing to Disclose: EC, OT, NC, BK, AG, MS, IU, ET, BU, MS, TD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1447 174 1107 SUN-50 PO34-01 Sunday 902 2012


901 ENDO12L_SUN-51 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Breast Size Increment during Pregnancy and Breast Feeding in PCOS Mothers: A Follow-Up Study of a Randomized Controlled Trial on Metformin vs. Placebo Eszter Vanky, Jorgen Nordskar, Hermann Leithe, Anna Hjorth-Hansen, Marit P Martinussen, Sven Magnus Carlsen St Olav[apos]s Hospital, Trondheim University Hospital, Trondheim, Norway; Norwegian University of Science and Technology, Trondheim, Norway; University of Debrecen, Debrecen, Hungary; Unit for Applied Clinical Research, Institute for Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway [bold]Background[/bold]: The significance of breast size increment in PCOS women during pregnancy, and the impact of metformin on breast feeding duration are essentially unexplored.[bold][br]Objectives: [/bold]To study breast size increment in pregnancy and the impact of metformin during pregnancy on breast feeding in women with PCOS.[bold][br]Design: [/bold]A follow-up study of a randomized controlled multicentre trial (The PregMet study).[bold][br]Intervention:[/bold] Women with PCOS were randomized to metformin or placebo from first trimester to delivery. Questionnaires were sent to 256 participants one year postpartum, 199 responded. [bold][br]Main Outcome Measures:[/bold] Pre-pregnancy and late pregnancy bra size, breast feeding pattern were registered and androgen levels measured in the mothers. [bold][br]Results:[/bold] No difference in breast size increment and breast feeding were found between the placebo and metformin group. Breast size increment during pregnancy correlated positively to the duration of exclusive breastfeeding, while BMI correlated negatively to the duration of partial breast feeding. Dehydroepiandrosterone-sulfate (DHEAS), testosterone and free testosterone index (FTI) in pregnancy did not correlate to breast size increment or duration of breast feeding. Women with no change in breast size were older, more obese, had higher blood pressure, serum triglycerides and fasting insulin levels, and had shorter duration of breast feeding compared to those with breast size increment. [bold][br]Conclusions:[/bold] PCOS women with no breast size increment in pregnancy seem to be more metabolically disturbed and less able to breast feed. Metformin supplementation during pregnancy did not improve their capability to breast feed.[br][br]Nothing to Disclose: EV, JN, HL, AH-H, MPM, SMC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 396 174 1108 SUN-51 PO34-01 Sunday 903 2012


902 ENDO12L_SUN-52 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Essential Oils of [italic]Lavundula angustifolia[/italic] and Tea Tree Oil Quantitatively and Qualitatively Reduce Hair Distribution in Young Women Giancarlo Balercia, Lara Giovannini, Francesca Paggi, Giacomo Tirabassi, Giorgio Panin, Marco Boscaro Polytechnic University of Marche, Ancona, Italy; Hulka srl, Rovigo, Italy Introduction: Essential oils of Lavundula angustifolia (Lavundula officinalis) and Malaleuca alternifolia (called Tea Tree Oil) are widely used in cosmetics due to their antiseptic, anti-inflammatory and relaxing properties.[br]Recent studies have shown that their use produces a slight estrogenic and anti-androgenic activity and contributes to a [ldquo]disequilibrium[rdquo] in the signaling pathway of sex hormones (1). On the basis of this evidence, we assumed that a daily topical application of these essential oils on young hypertrichotic female could reduce hair distribution of skin. Aims: to verify the consistence (diameter and length) and hair density/cm[sup2] of skin after 3 months of daily application b.i.d (on the left forearm) of oil spray containing essential oils of Lavundula angustifolia and Tea Tree Oil, compared to no application (right forearm). Methods:12 young female volunteers (mean age 31.1[plusmn]4.3 years) with idiopathic hypertrichosis, not depilate for at least 6 months. They applied the oil spray (VEA Epil, Hulka S.r.l., Rovigo, Italy) b.i.d, morning and evening, on the back of the left forearm for 3 months. A self-assessment questionnaire of hair growth was given to the volunteers before and after treatment. The oil spray contained Cyclopentasiloxane, Tocopheryl Acetate, Tocopherol, Oryza Sativa/Oryza Sativa (rice) bran oil, Lavandula Angustifolia Oil, Melaleuca Alternifolia Oil. Results: after 3 months of daily treatment, the self-assessment questionnaire showed a qualitative (reduced consistency) and quantitative (hair density/cm[sup2] of skin) reduction of hair on the treated area in 80% of the studied subjects (10 women out of 12) and the efficacy continued even 3 months after treatment discontinuation. No patient had topical or systemic side effects. Conclusions: the use of essential oils of Lavundula angustifolia and Tea Tree Oil was found to be safe and effective in reducing both quantitatively and qualitatively hair distribution assuring success in its use in the cosmetic field.[br][br]1) DV Henley, N Lipson, K S Korach, C A Bloch, Prepubertal Gynecomastia Linked to Lavender and Tea Tee Oils, N Engl J Med 2007; 356:479-85.[br][br]Disclosures: GP: Consultant, Hulka s.r.l. Nothing to Disclose: GB, LG, FP, GT, MB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 179 174 1109 SUN-52 PO34-01 Sunday 904 2012


903 ENDO12L_SUN-53 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) The Relationship between Androgen Hormones and Mean Platelet Volume in Ovulatory Polycystic Ovary Syndrome Patients without Insulin Resistance Bercem Aycicek Dogan, Ayse Arduc, Mazhar Muslum Tuna, Yasemin Tutuncu, Iffet Dagdelen, Yavuz Yalcin, Dilek Berker, Serdar Guler Ankara Numune Training and Research Hospital, Ankara, Turkey OBJECTIVE[br]The relationship between metabolic with hormonal features of polycystic ovary syndrome (PCOS) and several risk factors of cardiovascular disease (CVD) have demonstrated in patients with PCOS. Mean platelet volume (MPV) is generally accepted as a new risk marker of CVD. Our study was designed to determine association between Androgen hormones (AH) and MPV levels in ovulatuar PCOS patients without insulin resistance (IR).[br]METHODS[br]Sixty one patients with newly diagnosed ovulatuar PCOS (regarding to criteries of Rotterhdam 2003) who were non-obese, without IR and 40 control healty subjects were included in our study. Subjects were excluded who had several chronic diseases and taken any medication which could affect value of MPV. Venous blood samples was taken for total blood count (including MPV), total testosterone (TT), free testosterone (FT), dehydroepiandrosterone-sulfate (DHEAS) and androstenedione (A) levels. Insulin resistance was calculated from blood chemistry measurements of fasting insulin and glucose according to updated homeostasis model assessment (HOMA-IR) method (fasting glucose(mg/dl) x serum insulin([micro]IU/ml)/405) and was considered significant value of 2.7 and higher results.[br]RESULTS[br]There was no difference between the groups regarding to age, BMI and HOMA-IR (p[gt]0.05, for all). TT, FT, A, DHEAS levels were higher in PCOS patients comparing with control group (p[lt]0.01, for all). No difference was observed in MPV values for two groups (p=0.521). There was no correlation between TT, FT, A, DHEAS with MPV(p[gt]0.05, for all).[br]DISCUSSION[br]The relationship between AH and MPV levels were unclear in PCOS patients. Whereas some small studies had show that correlation between DHEAS and MPV in PCOS patients (who were obese and with IR), our study did not. Our study indicated that hiperandrojenemia was not elevate MPV in ovulatuar PCOS patients, like the other prospective studies which were done with the other risk markers of CVD.[br][br]Nothing to Disclose: BAD, AA, MMT, YT, ID, YY, DB, SG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 619 174 1110 SUN-53 PO34-01 Sunday 905 2012


904 ENDO12L_SUN-54 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Effect of an Oral Contraceptive on Basal and Postprandial Ghrelin, PYY, CCK and Satiety in Lean Women with Polycystic Ovary Syndrome Gulcan Arusoglu, Gulden Koksal, Nese Cinar, Serkan Tapan, Duygu Yazgan Aksoy, Bulent Okan Yildiz Hacettepe University, Ankara, Turkey; Hacettepe University, Ankara, Turkey; Gulhane School of Medicine, Ankara, Turkey; Etlik Ihtisas Research and Training Hospital, Ankara, Turkey [bold]BACKGROUND: [/bold]Ghrelin, peptide YY (PYY), and cholecystokinin (CCK) are gut hormones involved in appetite regulation and energy homeostasis, and are biological neuroendocrine signals that potentially affect food intake and adiposity. Ghrelin is an orexigenic peptide that stimulates food intake whereas PYY and CCK are anorexigenic peptides that might antagonize the effects of ghrelin.[br][bold]OBJECTIVE: [/bold]We aimed to determine whether fasting or postprandial ghrelin, PYY, CCK and satiety responses are different between lean patients with PCOS and healthy women. We also aimed to assess potential impact of oral contraceptive use on these appetite regulating hormones and subjective appetite in PCOS.[br][bold]METHODS: [/bold]Eighteen lean PCOS patients and 18 healthy control women matched for age, BMI, body fat %, waist circumference (WC) and waist to hip ratio (WHR) (22.1[plusmn]4.2 years; 22.2[plusmn]3.3 kg/m[sup]2[/sup]; 25.9[plusmn]6.7%; 75.2[plusmn]8.5 cm; and 0.76[plusmn]0.06 for PCOS respectively) underwent for measurements of circulating ghrelin, PYY, CCK, and subjective appetite after a standardized mixed meal (300 kcal: 16% protein, 29.2% fat and 53.8% carbohydrate) at 0, 15, 30, 45, 60, 90, 120, and 180 minutes. The measurements were repeated in PCOS patients after 3 months of treatment with ethinyl estradiol 30 mcg/drospirenone 3 mg (EE/DRSP).[br][bold]RESULTS:[/bold] At baseline, fasting ghrelin, PYY, CCK and area under the curve (AUC) values of PYY, CCK, and Satiety Index (SI) were similar between PCOS patients and controls whereas PCOS patients had significantly lower AUC values for ghrelin compared to controls (p=0.04). Fasting and AUC values of ghrelin, PYY, CCK and SI did not show a significant change after treatment with EE/DRSP for 3 months.[br][bold]CONCLUSIONS:[/bold] Our results suggest that lean patients with PCOS compared to body fat-matched healthy women have similar satiety and concentrations of anorexigenic peptides PYY and CCK both at fasting and postprandial state. Even though fasting ghrelin levels are similar between the two groups, PCOS patients have more ghrelin suppression in response to a meal. Oral contraceptive use for 3 months does not alter satiety or concentrations of fasting and postprandial ghrelin, PYY and CCK.[br][br]Nothing to Disclose: GA, GK, NC, ST, DYA, BOY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 624 174 1111 SUN-54 PO34-01 Sunday 906 2012


905 ENDO12L_SUN-55 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Role of microRNA in PCOS: microRNA-93 Over-Expression Induces IL6 in Adipocytes Saleh Gamal Heneidi, Yen-Hao Chen, Gloria Mabel Gamboa, Ricardo Azziz Georgia Health Sciences University, Augusta, GA; Georgia Health Sciences University, Augusta, GA; Georgia Health Sciences University, Augusta, GA Rationale: Dysfunctional glucose metabolism and/or action of adipokines in adipose tissue contributes significantly to the insulin resistance (IR) and hyperinsulinemia of polycystic ovary syndrome (PCOS).[br]Background: Adipose tissue (AT) is the largest endocrine organ in the body and is metabolically active, secreting adipokines, including IL6, thereby modulating insulin action and controlling systemic insulin sensitivity and metabolic homeostasis. PCOS has varied levels of multiple cytokines, including higher levels of circulating IL6. PCOS AT also has higher numbers of resident macrophages, which may result in altered secretion of macrophage-associated cytokines. The insulin sensitive glucose transporter, GLUT4, expression is significantly lower in PCOS vs. matched controls. In human adipose tissue we observed significant negative associations between both GLUT4 expression and HOMA-IR. MicroRNA (miRNA) are post-transcriptional regulators that bind to complementary sequences on target mRNAs, often resulting in translational repression or target degradation and gene silencing. miRNAs regulate numerous biological mechanisms. In our previous microarray studies, miRNA arrays indicate PCOS adipose to have a differentially expressed miRNA profile, with microRNA-93 (miR-93) being upregulated in adipocytes from women with the disorder.[br]Objectives: To determine the effects of miR-93 on IL6 and glucose metabolism in human adipocytes.[br]Results: Both human and mouse 3T3-L1 adipocytes treated with IL6 exhibit significantly reduced expression of GLUT4. IL6-induced GLUT4 reduction correlates with a decrease in the transcription factors PPAR-[gamma] and SREBF-1, and an increase in CREB-[alpha]. In human adipocyte culture, the over-expression of miR-93 significantly increased intracellular IL6 gene expression at 48 hours. Additionally, over-expression of miR-93 significantly increased IL6 in the medium at 48 hours. In 3T3-L1 cells, over-expression of miR-93 significantly increased intracellular IL6 gene expression at 24 hours.[br]Conclusions: Our findings indicate that induction of miR-93 stimulates IL6 secretion in adipocytes, which in turn may contribute to decreased GLUT4 expression. Thus, miR-93 over-expression may contribute to the decreased glucose metabolism observed in PCOS adipocytes. The affects of miR-93 on adipokine production and secretions is a novel mechanism, and may play a role in IR in PCOS adipocytes.[br][br]Sources of Research Support: NIH grant RO1-DK073632 awarded to R.A.[br][br]Nothing to Disclose: SGH, Y-HC, GMG, RA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2263 174 1112 SUN-55 PO34-01 Sunday 907 2012


906 ENDO12L_SUN-56 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Reduced Expression of SIRT1 and SIRT3 in the Skeletal Muscle of PCOS Women Jeffrey D Covington, Eric Ravussin, Leanne M Redman Pennington Biomedical Research Center, Baton Rouge, LA Sirtuin 1 (SIRT1) and Sirtuin 3 (SIRT3) are deacetylases that protect against cellular stress and are associated with healthy metabolic function. Recently, SIRT1 has been shown to interact with and influence the function of the Androgen Receptor (AR). SIRT3 has been shown to localize to mitochondria in response to cellular stress to preserve mitochondrial function. To our knowledge, the roles of SIRT1 and SIRT3 in polycystic ovary syndrome (PCOS), an endocrine condition characterized by hyperandrogenemia and insulin resistance, has not been studied. We hypothesized that SIRT1 and SIRT3 would be reduced in skeletal muscle, the principle insulin responsive tissue. Skeletal muscle biopsies were obtained from twelve overweight and obese women with PCOS and ten normally menstruating controls, matched for age and BMI. SIRT1 gene expression was 41% lower (p = 0.003) in PCOS compared to controls with no change in AR gene expression (p = 0.92). The gene expression of Hypermethylated in Cancer 1 (HIC1), a transcriptional repressor that is activated in response to cellular stress and interacts with the SIRT1 promoter, was 11 fold higher (p = 0.0005) in PCOS vs. controls. Furthermore, SIRT3 gene expression was 44% lower (p = 0.001) in women with PCOS vs. controls. This study provides the first evidence for a potential role of SIRT1 in the molecular dysregulation of the androgen receptor in women with PCOS. In addition, this data suggests that PCOS women may be more susceptible to mitochondrial and metabolic dysfunction due in part to reduced SIRT3 expression in skeletal muscle.[br][br]Nothing to Disclose: JDC, ER, LMR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 681 174 1113 SUN-56 PO34-01 Sunday 908 2012


907 ENDO12L_SUN-57 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Role of microRNA in PCOS: miR-93 Overexpression Regulates GLUT4 Yen-Hao Chen, Saleh Heneidi, G Mable Gamboa, Ricardo Azziz Georgia Health Sciences University, Augusta, GA; Georgia Health Sciences University, Augusta, GA Rationale: Dysfunctional glucose metabolism in adipose tissue contributes significantly to the insulin resistance (IR) and hyperinsulinemia of polycystic ovary syndrome (PCOS) and type 2 diabetes mellitus (T2DM).[br]Objectives: To explore the role of miRNAs in PCOS and determine whether miRNA has a role in regulating the insulin sensitive glucose transporter (GLUT4) in human adipocytes.[br]Findings: Analysis of various signaling intermediaries of the IRS/PI3K/AKT pathway did not identify notable defects in PCOS adipocytes. However, GLUT4 expression is significantly lower in PCOS vs. matched controls. In human adipose tissue we observed significant negative associations between both GLUT4 expression and HOMA-IR. Additionally, miRNA arrays indicate PCOS adipose to have a differentially expressed miRNA profile, with microRNA-93 (miR-93) being upregulated in adipocytes from women with the disorder. The induced overexpression of miR-93 in cultured human and mouse 3T3-L1 adipocytes resulted in down-regulated GLUT4 gene expression. In addition, in human adipose tissue we observed significant negative associations between miR-93 and GLUT4 expression.[br]Conclusions: These results indicate that miR-93 expression is upregulated in women with PCOS, and that this upregulation is associated with decreased glucose metabolism. These results suggest miR-93 may directly inhibit GLUT4 expression. This appears to be a novel mechanism for IR in PCOS adipocytes.[br][br]Sources of Research Support: NIH Grant RO1-DK073632 awarded to R.A.[br][br]Nothing to Disclose: Y-HC, SH, GMG, RA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1980 174 1114 SUN-57 PO34-01 Sunday 909 2012


908 ENDO12L_SUN-58 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Carrier Frequency of Congenital Adrenal Hyperplasia [italic]CYP21A2[/italic] Variants in Cyprus Vassos Neocleous, Alexia AP Phedonos, Christos Shammas, Meropi Toumba, Charilaos Stylianou, Elena Andreou, Michalis Picolos, Tassos C Kyriakides, Nicos Skordis, Leonidas A Phylactou The Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus; Makarios III Hospital, Nicosia, Cyprus; Paphos General Hospital, Paphos, Cyprus; Makarios III Hospital, Nicosia, Cyprus; Alithias Center, Nicosia, Cyprus; Yale University, New Haven, CT Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency (21-OHD) is the most common autosomal recessive disorder and is mostly attributable to mutations in the [italic]CYP21A2[/italic] gene. The carrier frequency of [italic]CYP21A2[/italic] mutations in the general population has been estimated to be 1:25 to 1:10. In North America and Europe the prevalence of the severe classic form of CAH is estimated between 1:10,000 to 1:15,000 live births while the milder Non Classic CAH occurs in approximately 0.2% of the general population. So far the true carrier frequency for CAH due to 21-OHD has not been determined by comprehensive mutation analysis of the [italic]CYP21A2 [/italic]in a specific European population. The present study screened for mutations in the [italic]CYP21A2[/italic] gene a statistically valid number of 300 clinically asymptomatic subjects (150 males and 150 females) from the general population of Cyprus. The methodology used for the [italic]CYP21A2[/italic] genotyping involved multiplex ligation-dependent probe amplification (MLPA) and direct sequencing of PCR products of the [italic]CYP21A2 [/italic]gene. Genotyping of the 600 unrelated alleles from the 300 Cypriot asymptomatic individuals revealed a carrier frequency of 9.83%. The most frequent mutations among the tested subjects of the present study were the mild p.V281L (4.3%), followed by p.Q318stop (2.5%), p.P453S (1.33%), p.V304M (0.83%), p.P482S (0.67%) and p.M283V (0.17%). In conclusion, the detected 9.83% CAH carrier frequency suggests one of the highest prevalence of [italic]CYP21A2[/italic] carriers reported by a genotyping analysis and the previously described major mutations are found to dominate the mutation spectrum of the Cypriot population. In addition, the rare V304M mutation which to our knowledge was reported only once before in a female patient of Asian origin seems to be quite frequent (0.83%) in the Cypriot asymptomatic population and imply a possible founder effect. Knowing of the prevalence and the nature of the genetic defects in our population will be of immense help in our understanding and awareness of NC CAH in females presenting with hyperandrogenemia.[br][br]Nothing to Disclose: VN, AAPP, CS, MT, CS, EA, MP, TCK, NS, LAP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 223 174 1115 SUN-58 PO34-01 Sunday 910 2012


909 ENDO12L_SUN-59 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Prenatal Testosterone Excess Decreases FSH Levels in Umbilical Cord Blood and Modifies the Expression of Key Reproductive Factors in Testis of Fetal Male Lambs Pedro Pablo Rojas-Garcia, Monica Patricia Recabarren, Teresa Sir-Petermann, Rodolfo Rey, Albert Carrasco, Romina Fornes, Sergio Edmundo Recabarren Universidad de Concepcion, Chillan, Chile; Western Medical School, Universidad de Chile, Santiago, Chile; University of Buenos Aires, Buenos Aires, Argentina The programming effects of prenatally exposure to excess testosterone (T) on postnatal reproductive, metabolic, and behavioral parameters have been studied extensively in females of several species but similar studies in males are limited. We have previously found that prenatal T treatment leads to an increased Sertoli cell number but reduced germ cell number, altered spermatogenesis and increased FSHR expression in adult rams. If such disruptions are established early during fetal development because of the changes of fetal endocrine environment due to T treatment remain to be explored. This study addressed the impact of prenatal T excess on hormone levels and expression of genes modulating testicular function in fetuses of 120 days. Pregnant Suffolk sheep were administered either testosterone propionate 30 mg (from day 30 to 90) followed by 40 mg (from day 90 to 120) i.m. twice weekly of pregnancy (term is [sim]147 days) or vehicle. At 120 days of pregnancy, dams were sedated and subsequently maintained under general anesthesia. The gravid uterus was exposed to obtain blood from the umbilical artery. Fetuses were removed for body measures and were given an intracardiac barbiturate prior to tissue harvest. Hormone levels were measured using radioimmunoassay and the mRNA expression was measured using real time PCR. Body and testicular weight were similar between males fetuses born to dams treated with T propionate (T-males, n=5, 2.30 [plusmn] 0.26 Kg, body weight and 0.317 [plusmn] 0.05 g testicular weight) and control fetuses born to dams treated with vehicle (C-males, n=8, 1.99 [plusmn] 0.06 Kg body weight and 0.336 [plusmn] 0.02 g testicular weight). T-males showed lower FSH levels in umbilical cord arterial blood than C-males (mean plasma levels were 0.2440 [plusmn] 0.083 ng/ml in T-males and 1.033 [plusmn] 0.16 ng/ml in C-males). In T-males, the mean T levels were 0.04 [plusmn] 0.014 ng/ml. T levels were below the detection limit of the assay in 5 of 8 C-males. The mean T concentrations in those with detectable T levels were 0.02 [plusmn] 0.006 ng/ml. Estradiol levels were 122.5 [plusmn] 52.82 in C-males, but in T-males only 3 fetuses had detectable levels in which the mean was 20.45 [plusmn] 7.59 pg/ml. In testicular tissue FSHR, AR, AMH, TGFB3 and TGFBR1 expressions were higher in T-males than C-males. These findings provide evidence that consequences of fetal exposure to excess T are manifested already during fetal development and may impact the testicular function during the adulthood of male rams.[br][br]Sources of Research Support: FONDECYT Grant 1090031.[br][br]Nothing to Disclose: PPR-G, MPR, TS-P, RR, AC, RF, SER 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1337 174 1116 SUN-59 PO34-01 Sunday 911 2012


910 ENDO12L_SUN-60 POSTER SESSION: Hyperandrogenic Disorders (1:30 PM-3:30 PM) Prenatal Exposure to Excess Testosterone Modifies the Feedback to Testosterone in Ovariectomized Adult Female Sheep Monica Patricia Recabarren, Pedro Pablo Rojas-Garcia, Albert Carrasco, Natalia Parra, Teresa Sir-Petermann, Sergio Edmundo Recabarren Universidad de Concepcion, Chillan, Chile; Western Medical School, Universidad de Chile, Santiago, Chile Prenatal exposure to testosterone (T) in female sheep causes abnormalities in the neuroendocrine axis including hypersecretion of LH. These alterations suggest a defeminization of the neuroendocrine axis. The mechanisms that mediate the LH hypersecretion in these females involve increased sensibility to GnRH, increase in the expression of receptors to GnRH in the pituitary and decrease in the expression of estrogen receptors. However, the evaluation of the sensibility of these females to T has not been completely explored, considering that these females may show a defeminizated feedback pattern. Therefore, we evaluated the T feedback to LH and the LH response to a GnRH agonist after an acute treatment with T in prenataly treated OVX ewes. At 40 weeks of age we assessed the LH and FSH episodic secretion by collecting blood samples from the jugular vein every 10 minutes for 6 hours. Thereafter, we determined the LH plasma concentrations in response to a bolus of GnRH agonist by collecting blood samples every 30 min for the first 3 hours and every 3 and 6 to complete 48 hours. Both experiments were performed before and 48 h after an acute testosterone propionate (TP, 40mg/sheep) challenge. We used 6 Suffolk-Down ewes born to dams exposed to 30 mg TP (T-females group, n=6) between 30-90 days of pregnancy, followed by 40 mg between 90-120 days of pregnancy. Females born to a vehicle treated mothers were control (C-females, n=7). Before the T challenge, plasma T concentrations were undetectable in both groups. Parameters of LH pulsatility were similar between both groups. In C-females, the T challenge caused a significant decrease in the number of LH pulses and an increase in mean LH secretion, LH pulse amplitude and nadir. In the T-females, the T challenge did not modify any of the parameters of the episodic LH secretion except for a decrease in the pulse frequency. Between groups, the T challenge elicited a significant decrease in mean LH secretion, in LH pulse amplitude and nadir in T-females in comparison to C-females. FSH pulsatility characteristics were not different within or between groups. LH secretion after the GnRH challenge, reached a peak within the first 3 hours, however, no significant differences between groups were observed. Results suggest that fetal exposure to T induces a defeminization of the negative feedback to T in female sheep.[br][br]Sources of Research Support: Fondecyt Grant 1090031.[br][br]Nothing to Disclose: MPR, PPR-G, AC, NP, TS-P, SER 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1327 174 1117 SUN-60 PO34-01 Sunday 912 2012


911 ENDO12L_SUN-61 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Leydig Cell Hyperplasia Is the Predominant Testicular Phenotype in McCune-Albright Syndrome Alison M Boyce, William H Chong, Thomas A Shawker, Peter A Pinto, W Marsten Linehan, Maria J Merino, Frederick R Singer, Michael T Collins National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; John Wayne Cancer Institute, Santa Monica, CA The male gonadal phenotype in McCune-Albright syndrome (MAS) has not been well characterized. Due to the comparatively low prevalence of precocious puberty (PP) in boys, common thinking is that males with MAS have a significantly lower prevalence of gonadal pathology than females. Methods: Biochemical, radiologic and histologic analyses were performed in males with MAS as part of an ongoing prospective cohort study. Analyses included physical exam, testicular ultrasound (TUS), LH, FSH and testosterone. Surgical procedures were performed when clinically indicated. Results: 121 subjects with MAS were evaluated, including 54 males (44%) (3-59y). 44 (81%) had TUS abnormalities including solid lesions (49%), cystic lesions (30%), microlithiasis (30%), heterogeneity (47%), and focal calcifications (11%). TUS lesions were extensive with features concerning for malignancy, and 11 subjects underwent unilateral total or partial orchiectomy. Pathology in all subjects showed extensive nodular Leydig cell hyperplasia (LCH), while one subject had Sertoli cell hyperplasia in addition. Pathologic exam could not exclude malignant Leydig cell tumor. No evidence of local invasion was found intra-operatively or on histology, and CT imaging showed no metastases. Post-operatively subjects were followed with serial surveillance imaging, and after none developed clinical malignancy management was altered to reflect a lower likelihood of malignant transformation. Subsequent subjects were followed with imaging alone, and with longitudinal observation (mean 4.6 years, range 1-13) no cases of malignancy have developed. Unilateral or bilateral macro-orchidism was present in 44% with no discrete palpable masses. Testicular function appeared to be preserved, with normal testosterone and gonadotropins in subjects without PP or pituitary disease. PP occurred in 83% of females, equivalent to the prevalence of testicular abnormalities in males. Conclusion: We have characterized a novel gonadal phenotype in the largest reported cohort of males with MAS. TUS should be performed in all males evaluated for MAS, and characteristic abnormalities should be considered a diagnostic criterion. Risk of malignant transformation is likely low, and we advocate a conservative approach to management with emphasis on testicular preservation, close observation and serial imaging.[br][br]Sources of Research Support: The Intramural Research Program of the NIH, NIDCR and NICHD.[br][br]Nothing to Disclose: AMB, WHC, TAS, PAP, WML, MJM, FRS, MTC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 888 175 1118 SUN-61 PO36-02 Sunday 913 2012


912 ENDO12L_SUN-62 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Insulin-Like Peptide 3 (INSL-3) Levels in Prader-Willi Syndrome (PWS) Males from Infancy through Adulthood Harry J Hirsch, Varda Gross-Tsur, Najiba Lahlou, Marc Roger, Talia Eldar-Geva Shaare Zedek Medical Center, Jerusalem, Israel; Hopital Cochin, Paris, France; Hopital Saint Vincent de Paul, Paris, France; Shaare Zedek Medical Center, Jerusalem, Israel [bold]Background:[/bold] Cryptorchidism, incomplete pubertal development, and low serum testosterone levels are manifestations of hypogonadism seen in nearly all PWS males. Hypogonadism in PWS results from variable combinations of hypothalamic and primary testicular dysfunction (1). The Leydig cell hormone, INSL-3, facilitates testicular descent in the male fetus. Levels continue to rise in late adolescence but the functional role of INSL-3 in adults is not known. INSL-3 levels in PWS males have not been reported. [bold]Objectives:[/bold] Characterize changes in INSL-3 with age in PWS boys and men and examine the relation of INSL-3 to LH, FSH, testosterone (T), anti-Mullerian hormone (AMH) and inhibin B. [bold]Study design[/bold]: We obtained blood samples from 41 PWS males (24 DEL, 17 UPD), ages 12.9[plusmn]10.1 years (2 months to 36 years). In 12 patients, a second blood sample was obtained 1.8[plusmn]0.4 years (1.4 to 2.7 years) after the initial sample. Hormones were measured by immunoassay (1,2). All but one had cryptorchidism (bilateral in 28). Mean age at orchiopexy was 3.0[plusmn]0.8 years. [bold]Results:[/bold] For age groups [lt] 1 year, 1-12 years, 12-18 years, and [gt]18 years, mean[plusmn]SD levels of INSL-3 were 176[plusmn]134, 101[plusmn]206, 285[plusmn]334, and 604[plusmn]349 pg/ml, respectively. INSL-3 showed positive correlations (Spearman Rho) with LH (p[lt]0.0001) and T (p[lt]0.0001), but not with FSH, AMH, or inhibin B. INSL-3 were similar in boys with unilateral vs bilateral cryptorchidism. Among 18 patients above age 14 years, 5 had low INSL-3 levels (39.2[plusmn]18.7 pg/ml) along with low LH and FSH (0.4[plusmn]0.6 and 1.4[plusmn]1.6 mIU/ml). The other 13 men in these age group had normal INSL-3 (701.1[plusmn]252.8 pg/ml), normal LH (3.6[plusmn]2.5 mIU/ml), and elevated FSH (18.6[plusmn]17.9 mIU/ml). Significant differences in LH (p[lt]0.0008) and FSH (p[lt]0.005) were observed for the low vs normal INSL-3 groups, but no significant differences were seen for testosterone, AMH, or inhibin B. Longitudinal data confirm the decrease in INSL-3 from high values in infancy, to low levels during childhood, followed by an increase to adult levels in later adolescence. [bold]Conclusions: [/bold]INSL-3 levels in most PWS males are in the expected normal range and follow age-related patterns. INSL-3 levels were low in the gonadotropin-deficient adult men, but were normal in most adults; in contrast to serum T which was low in all PWS boys and men. These results suggest that a specific defect in testosterone synthesis, rather than a general impairment of Leydig cell function contributes to the hypogonadism in PWS males.[br][br](1) Hirsch HJ et al., J Clin Endocrinol Metab 2009; 94: 2262. (2) Cabrol S et al., J Clin Endocrinol Metab 2011; 96: E746.[br][br]Sources of Research Support: Foundation for Prader-Willi Research; Pfizer Pharmaceuticals.[br][br]Nothing to Disclose: HJH, VG-T, NL, MR, TE-G 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1889 175 1119 SUN-62 PO36-02 Sunday 914 2012


913 ENDO12L_SUN-63 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Inhibition of Steroidogenesis in Normal Men with Experimentally Induced Hypogonadotropic Hypogonadism Maximally Decreases Intratesticular Testosterone Mara Y Roth, Kat Lin, Stephanie T Page, Jean-Jacques S Nya-Ngatchou, Bradley D Anawalt, Alvin M Matsumoto, Brett T Marck, William J Bremner, John K Amory University of Washington, Seattle, WA; University of Washington, Seattle, WA; University of Washington, Seattle, WA; Veterans Affairs Puget Sound Health Care System, Seattle, WA Background: Intratesticular testosterone (ITT) is essential for spermatogenesis, yet spermatogenesis can persist despite near-complete suppression of gonadotropins and subsequent severe suppression of ITT. It is unknown whether a minimum ITT concentration exists for men, below which spermatogenesis ceases. We experimentally induced low levels of gonadotropins and ITT in normal men with the gonadotropin releasing hormone (GnRH) antagonist, acyline, then assessed the effect on ITT of adding an inhibitor of testosterone synthesis (ketoconazole), 5 [alpha]-reductase inhibitor (dutasteride, to lower DHT) or aromatase inhibitor (anastrazole, to lower estradiol).[br]Methods: Forty healthy men completed all study procedures. All men received the GnRH antagonist, acyline (300 mcg/kg SC once), and 1% testosterone (T) gel 5gm daily for 10 days (to maintain normal serum T). After 48 hours, subjects were randomized to one of five treatment groups: placebo (acyline alone), ketoconazole 400mg, ketoconazole 800mg, dutasteride 2.5mg, or anastrazole 1mg daily for 7 days. ITT and corresponding serum T were measured 48 hours after receiving acyline and at the end of drug-treatment on day 10. T was quantified using liquid chromatography with tandem mass spectrometry.[br]Results: Median ITT (25th, 75th percentile) in all subjects after receiving acyline was reduced to 105 (73, 135) nmol/L. After ten days of treatment, ITT decreased further in both groups receiving ketoconazole, to 13 (9, 25) nmol/L in the ketoconazole 400mg dose-group and to 6 (3, 14) nmol/L in the ketoconazole 800mg dose-group (p[lt]0.001 vs acyline alone). Subjects receiving placebo, dutasteride and anastrazole had no significant change in ITT on treatment. Normal serum T was maintained in all groups.[br]Conclusion: Inhibition of steroidogenesis by ketoconazole in gonadotropin-suppressed men markedly decreased ITT to a greater degree than with gonadotropin suppression alone. Administration of the combination of ketoconazole plus acyline to maximally suppress ITT will allow for future studies designed to examine the impact of extremely low ITT on normal human spermatogenesis.[br][br]Sources of Research Support: Eunice Kennedy Shriver National Institute of Child Health and Human Development U54 HD-42454; Eunice Kennedy Shriver National Institute of Child Health and Human Development grant K12 HD053984 to support MYR.[br][br]Nothing to Disclose: MYR, KL, STP, J-JSN-N, BDA, AMM, BTM, WJB, JKA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 362 175 1120 SUN-63 PO36-02 Sunday 915 2012


914 ENDO12L_SUN-64 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Pulsatile Infusion of Recombinant Human LH (rhLH) in GnRH Antagonist-Treated Men Stimulates Testosterone (T) Secretion More Than Continuous LH Infusion Paul Y Takahashi, Daniel M Keenan, Johannes D Veldhuis Center for Translational Science Activities, Rochester, MN; University of Virginia, Charlottesville, VA [underline]Background[/underline]. In the rat and ram, pulsatile and continuous LH infusions achieve comparable T output (1,2). No similar studies exist in humans. However, rapid testicular desensitization occurs after (continuous) hCG stimulation in men (3).[br][underline]Hypothesis[/underline]. Pulsatile i.v. infusions of rhLH in acutely T-depleted men will restore T secretion more effectively than continuous i.v. delivery of the same total LH dose.[br][underline]Methods[/underline]. Twenty healthy men were each studied 4 times in prospectively randomized double-blind order overnight. A maximally inhibitory dose of ganirelix, a GnRH antagonist, was given at 2200 hr in each subject, followed by:[br]1) i.v. saline infusion for 12 hr (zero rhLH addback)[br]2) continuous i.v. rhLH infusion over 12 hr (112.5 units rhLH total)[br]3) 18.75 units rhLH i.v. bolus every 2 hr over 12 hr (112.5 units rhLH total), and[br]4) 9.4 units rhLH i.v. bolus every 1 hr over 12 hr (112.5 units rhLH total).[br]A standardized pulsatile stimulus was applied at the end of the overnight 12 hr comprising rhLH 37.5 units i.v. over 6 min every 2 hr for 3 pulses. LH and T were measured in 10-min serum samples collected over the last 16 hr of all 80 infusions. LH-T dose-response estimation was applied to the 16-hr data (4).[br][underline]Outcomes[/underline]. Mean ([plusmn] SEM) total T concentrations (ng/dL) over the 12-hr infusion interval after ganirelix administration were 93 [plusmn] 35 (saline), 190 [plusmn] 73 (continuous rhLH addback), 313 [plusmn] 110 (rhLH 2-hr pulses) and 320 [plusmn] 96 (rhLH 1-hr pulses) [P[lt]10[sup]-13[/sup]]. The rank order of T responses was 1-hr = 2-hr rhLH pulses [gt] continuous rhLH [gt] saline infusions. Corresponding LH concentrations (IU/L) were 0.88 [plusmn] 0.31, 1.8 [plusmn] 0.69, 3.7 [plusmn] 1.2 and 4.2 [plusmn] 1.6. During the terminal standardized (3-consecutive) rhLH pulses, T rose to 236 [plusmn] 68 (saline), 274 [plusmn] 65 (continuous), 324 [plusmn] 99 (2-hr pulses) and 343 [plusmn] 75 (1-hr pulses) [P=0.0002], with a rank order of treatment effects: 1-hr = 2-hr [gt] continuous [gt] saline pulses. By nonlinear dose-response estimation, the ED[sub]50[/sub] for infused LH was significantly lower for 2-hr than 1-hr pulses, [italic]viz[/italic]: 2.9 IU/L [italic]vs[/italic] 4.9 IU/L (P=0.034), defining greater apparent potency of 2-hr pulses.[br][underline]Conclusions[/underline]. Continuous rhLH infusion is less stimulatory than rhLH pulses, but more effective than overnight saline under GnRH antagonism, showing for the first time that human Leydig cells depend upon the time-mode of lutropic drive.[br][br]1. Gibson-Berry KL, Chase DJ 1990 Continuous and pulsatile infusions of luteinizing hormone have identical effects on steroidogenic capacity and sensitivity of Leydig cell in rats passively immunized against gonadotropin-releasing hormone. Endocrinol 126(6):3107-3115. 2. Chase DJ, Schanbacher BD, Lunstra DD 1988 Effects of pulsatile and continuous luteinizing hormone (LH) infusions on testosterone responses to LH in rams actively immunized against gonadotropin-releasing hormone. Endocrinol 123(2):816-826. 3. Glass AR, Vigersky RA 1980 Resensitization of testosterone production in men after human chorionic gonadotropin-induced desensitization. J Clin Endocrinol Metab 51:1395-1400. 4. Keenan DM, Iranmanesh A, Veldhuis JD 2011 Analytical construct of reversible desensitization of pituitary-testicular signaling: illustrative application in aging. Am J Physiol Regul Integr Comp Physiol 300(2):R349-350.[br][br]Sources of Research Support: In part via the Center for Translational Science Activities (CTSA) Grant Number 1 UL 1 RR024150 from the National Center for Research Resources (Rockville, MD), and AG019695, DK073148, AG029362, AG031763 and DK050456 (Metabolic Studies Core of the Minnesota Obesity Center) from the National Institutes of Health (Bethesda, MD).[br][br]Nothing to Disclose: PYT, DMK, JDV 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 317 175 1121 SUN-64 PO36-02 Sunday 916 2012


915 ENDO12L_SUN-65 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) A New Combination of Testosterone and Nestorone[reg] Transdermal Gels for Male Hormonal Contraception Niloufar Ilani, Mara Y Roth, John K Amory, Ronald S Swerdloff, Clint Dart, Stephanie T Page, William J Bremner, Regine Sitruk-Ware, Narender Kumar, Diana Blithe, Christine Wang Harbor-UCLA Medical Center and Los Angeles Biomedical Research Institute, Torrance, CA; University of Washington, Seattle, WA; Agile Clinical Development, Durham, NC; Population Council, New York, NY; National Institutes of Health, Bethesda, MD [bold]Context: [/bold]Combinations of testosterone (T) and progestin transdermal gels enhance the suppression of spermatogenesis and prove effective and appealing to a majority of men for contraception. Provider-independent application of this regimen may increase acceptability and compliance among users.[br][bold]Objective: [/bold]To determine the effectiveness of T gel alone or combined with Nestorone (NES), a potent non-androgenic progestin, in gel formulation for suppressing spermatogenesis.[br][bold]Design and Setting:[/bold] Randomized, double-blind, comparator clinical trial conducted at two academic medical centers.[br][bold]Participants: [/bold]99 healthy male volunteers.[br][bold]Interventions: [/bold]Men were randomized to one of three treatment groups applying daily transdermal gels (Group 1: T 10g + NES 0mg/placebo gel; Group 2: T 10g + NES 8mg; Group 3: T gel 10g + NES 12mg).[br][bold]Outcome Variables[/bold]: 1) Percentage of men who reached a sperm concentration of [le]1 million/ml by 20-24 weeks of treatment. 2) The impact of each treatment on gonadotropins, sperm morphology and motility.[br][bold]Results: [/bold]Efficacy data analyses were performed on 56 subjects who adhered to the protocol and completed at least 20 weeks of treatment. The percentage of men with sperm concentration of [le]1 million/ml was significantly higher for T+NES 8 mg (89%, [italic]P[/italic] [lt]0.0001) and T+NES 12 mg (88%, [italic]P[/italic] [lt]0.0002) than T alone group (23%). In addition, significantly more men became azoospermic in the T+NES 8 (78%, [italic]P[/italic] [lt]0.001) and T+NES 12 (69%, [italic]P[/italic] [lt]0.008) groups compared to T+NES 0 (23%). The decrease in sperm motility and normal morphology followed the decrease in sperm concentration. Median serum luteinizing hormone and follicular stimulating hormone concentrations were more suppressed in the groups receiving NES together with T. Median total and free T concentrations remained within the adult male range irrespective of treatment group. All subjects recovered to a sperm concentration of [ge]15 million/ml during the recovery period. Adverse effects were minimal in all groups.[br][bold]Conclusion[/bold]: A combination of daily NES+T gels suppressed sperm concentration to [le]1 million/ml in the majority of men with minimal adverse effects. The results warrant further investigation of this combined male transdermal hormonal contraceptive regimen.[br][br]Sources of Research Support: NICHD Grant HHSN275200403369I, HHSN2751008060044U and the Population Council. The Los Angeles center was supported by: Endocrinology, Metabolism and Nutrition training Grant (T32 DK007571); General Clinical Research Center (MO1 RR00425) and the UCLA Clinical and Translational Science Institute (1UL1-RR033176) at Harbor-UCLA Medical Center and Los Angeles Biomedical Research Institute. The Seattle center was supported by: Eunice Kennedy Shriver NICHD Grant HHSN27520040337; Center for Research in Reproduction and Contraception U54 HD 04245 (NICHD); and Eunice Kennedy Shriver NICHD 5K12HD053984.[br][br]Nothing to Disclose: NI, MYR, JKA, RSS, CD, STP, WJB, RS-W, NK, DB, CW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 882 175 1122 SUN-65 PO36-02 Sunday 917 2012


916 ENDO12L_SUN-66 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Normalizing the Sex Hormone Balance Modulates Pro-Inflammatory Cytokine Secretion in Blood and Adipose Tissue in Men Tomas Ahern, Michelle Corrigan, Gadintshware Gaoatswe, Conor Woods, Cheryl Sweeney, Andrew Hogan, Elizabeth J Ryan, Donal O[apos]Shea, Frances J Hayes St Columcille[apos]s Hospital, Loughlinstown, Ireland; St Vincent[apos]s University Hospital, Elm Park, Ireland; St Vincent[apos]s University Hospital, Elm Park, Ireland; St Vincent[apos]s University Hospital, Elm Park, Ireland Introduction[br]Obese people have elevated serum pro-inflammatory cytokine concentrations, which may contribute to their increased risk of cardiovascular disease and cancer. Obesity in men is associated with low circulating testosterone (T) and high oestradiol (E[sub]2[/sub]) levels. Sex steroids have opposing effects on the secretion of pro-inflammatory cytokines from peripheral blood mononuclear cells (PBMCs), with T decreasing and E[sub]2[/sub] increasing their secretion. We therefore hypothesized that physiological concentrations of both T and E[sub]2[/sub] may favourably modulate pro-inflammatory cytokine secretion in blood and adipose tissue from men.[br]Methods[br]We exposed PBMCs from 20 obese men (BMI[gt]40kg/m[sup]2[/sup]) and adipose tissue stromavascular cells (SVCs) from 7 men undergoing laparoscopic abdominal surgery to varying levels of both T and E[sub]2[/sub] for 24 hours. We determined supernatant concentrations of tumour necrosis factor alpha (TNF[alpha]) and interleukin-6 (IL-6) by enzyme-linked immunosorbent assay. The T and E[sub]2[/sub] concentrations were chosen to reflect the following conditions: physiological (T 25nM, E[sub]2[/sub] 100pM); obese hypogonadal (T 5nM, E[sub]2[/sub] 150pM); and testosterone treated (T 25nM, E[sub]2[/sub] 200pM). Data are expressed as medians and as percentage of secretion by vehicle treated cells. Friedman and Wilcoxon Signed-Rank analyses were used to determine whether differences existed between sex hormone conditions.[br]Results[br]PBMC TNF[alpha] secretion was 95.2% under physiological conditions, 101.9% under obese hypogonadal conditions and 105.1% under T treated conditions (p=0.003). PBMC IL-6 secretion was 88.3% under physiological conditions, 97.4% under obese hypogonadal conditions and 100.3% under T treated conditions (p=0.02). SVC TNF[alpha] secretion was 95.3% under physiological conditions, 91.4% under obese hypogonadal conditions and 103.7% under T treated conditions (p=0.08 comparing all conditions, p=0.02 comparing physiological to T treated conditions).[br]Conclusions[br]Pro-inflammatory cytokine secretion from PBMCs and adipose tissue SVCs is lower in the presence of physiological T and E[sub]2[/sub] concentrations compared to the sex hormone milieu that is characteristic of a T-treated obese hypogonadal male. Inhibition of aromatase activity can normalise T and E[sub]2[/sub] levels in obese hypogonadal men. Aromatase inhibitor treatment is thus likely to have a more favourable effect on pro-inflammatory cytokine secretion than testosterone therapy and may therefore be the preferable therapeutic option for obese hypogonadal men.[br][br]Sources of Research Support: The Irish Heart Foundation and the Irish Health Research Board in the form of unrestricted research grants.[br][br]Nothing to Disclose: TA, MC, GG, CW, CS, AH, EJR, DO, FJH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1509 175 1123 SUN-66 PO36-02 Sunday 918 2012


917 ENDO12L_SUN-67 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Intratesticular 13-[italic]cis[/italic] Retinoic Acid Is Significantly Reduced in Men with Abnormal Spermatogenesis Compared to Men with Normal Spermatogenesis Jean-Jacques S Nya-Ngatchou, Sam Arnold, Thomas J Walsh, Charles Muller, Stephanie T Page, Nina Isoherranen, William J Bremner, John K Amory University of Washington, Seattle, WA; University of Washington, Seattle, WA; University of Washington, Seattle, WA [bold]Background[/bold]: Retinoic acid is necessary for spermatogenesis, but the tissue concentration of the bioactive retinoids, all-[italic]trans[/italic] and 13-[italic]cis[/italic] retinoic acid in men and their relationship to spermatogenesis have not been examined. We hypothesized that intratesticular concentrations of retinoids would be lower in men with impaired spermatogenesis as compared with control men.[br][bold]Subjects and Methods[/bold]: We recruited 30 men requiring scrotal surgery for varicocelectomy, hydrocelectomy or similar procedures in convenience sample to examine the relationship between spermatogenesis and intratesticular and serum retinoids. Twenty-five men completed two pre-operative measurements of serum hormones and retinoids, donated two specimens for semen analysis, and then had a testicular biopsy at the time of their previously planned surgery. Serum and tissue retinoids and hormones were measured by liquid chromatography/tandem mass spectrometry and specific radioimmunoassays, respectively.[br][bold]Results[/bold]: Seven men had abnormal spermatogenesis by at least one WHO criteria and 18 men were normal by all parameters. In men with abnormal spermatogenesis, the median (25[sup]th[/sup], 75[sup]th[/sup] percentile) intratesticular 13-[italic]cis[/italic]-retinoic acid was 0.18 (0.09, 0.25) pmol/gram tissue compared with 0.35 (0.23, 0.39) pmol/gram tissue in men with normal spermatogenesis (p=0.03). There was no significant difference in intratesticular all-[italic]trans[/italic] retinoic acid between these groups. There were no significant correlations between retinoids or hormones and parameters of spermatogenesis excepting a significant inverse correlation between the intratesticular-13 [italic]cis[/italic] retinoic acid and the serum all-[italic]trans[/italic] retinoic acid (r= -0.59, p=0.009)[br][bold]Conclusion[/bold]: Intratesticular 13-[italic]cis[/italic] retinoic acid is significantly lower in men with abnormal spermatogenesis compared to men with normal spermatogenesis. This difference may be a function of impaired biosynthesis or transport, or could be due to increased metabolism. Further investigation of the relationship between 13-[italic]cis[/italic] retinoic acid and spermatogenesis in men with infertility is warranted.[br][br]Sources of Research Support: This work was supported by the Eunice Kennedy Shriver National Institute of Child Health and Human Development, a division of the National Institute of Health through cooperative agreement U54 HD42454 as part of the Cooperative Contraceptive Research Centers Program.[br][br]Disclosures: STP: Principal Investigator, Abbott Laboratories. Nothing to Disclose: J-JSN-N, SA, TJW, CM, NI, WJB, JKA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 898 175 1124 SUN-67 PO36-02 Sunday 919 2012


918 ENDO12L_SUN-68 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Detection of Indirect Androgen Doping Using a GnRH Analog (Leuprolide) David J Handelsman, Amanda Idan, Catrin Goebel, Veena Jayadev, Leo Turner, Ann J Conway University of Sydney, Concord Hospital, Sydney, Australia; Concord Hospital, Sydney, Australia; National Measurement Institute, North Ryde, Sydney, Australia Indirect androgen doping (using drugs to increase endogenous androgen levels) has attracted attention following the prohibition of doping with exogenous androgens, which remain the most potent ergogenic drugs for power sports. While GnRH analog use is suspected, detection methods are not well developed.[br]We aimed to determine if (a) GnRH analog (leuprolide, L) stimulation of serum and urine LH and steroids is reproducible at a 2 week interval, (b) nandrolone decanoate (ND) co-administration influences responses to L, and (c) serum and urine LH and steroids (serum T, DHT, E2, E1, 3[alpha] [amp] 3[beta] diols; urine T, epitestosterone (E), androsterone (A)) levels provide an effective screening test.[br]Healthy men were randomized into a 4 week parallel group, open label clinical study. Leuprolide (1mg) was injected sc daily for 4 days in 1st [amp] 3rd week with hormone-free 2nd [amp] 4th weeks. In the 3rd week, men were randomized to either N decanoate injections or no extra treatment ([gt]99% scheduled injections [amp] samples collected, 18 cycles). Steroids (MS) [amp] LH (immunoassay) were assayed in batch with urine concentrations adjusted to SG 1.020. Mixed model ANOVA analysis used time, treatment (ND vs nil) [amp] genotype (CC, CJ, JJ) as main effects with age, height [amp] weight as covariates. UGT2B17 deletor genotyping was by duplex PCR defining wild-type (C) and mutant (J) alleles. Optimal cutpoint for classification was by Youden index in ROC analysis.[br]Leuprolide stimulated striking increases in serum and urine LH and steroids (serum T, DHT, 3[alpha] diol; urine T, E [amp] A) which were sustained for 4 days and of equal magnitude in both treatment periods (except serum DHT not increased in 2nd treatment). There was no response to L in serum DHEA, urinary T/E ratio or A/E ratio. ND suppressed basal serum T, E2, 3[alpha] diol, and urinary E but did not change the magnitude of the 2nd stimulation of LH and androgens. Homozygous UGT2B17 deletion genotype (JJ) produced extremely low urinary T and T/E ratio but with normal urinary E and all other measured steroids (including serum T). Optimal screening for L administration was provided by urinary log10(LH x T) which, using a cutpoint of 1.96, correctly classified 82% of samples. Improved discrimination may be achieved with an MS-based L assay under development.[br]We conclude that L stimulation of endogenous androgens is reproducible after a week treatment-free interval, is unaffected by co-administration of ND but could mask an ND-induced decrease in T/E ratio.[br][br]Sponsored by World Anti-Doping Agency.[br][br]Nothing to Disclose: DJH, AI, CG, VJ, LT, AJC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 844 175 1125 SUN-68 PO36-02 Sunday 920 2012


919 ENDO12L_SUN-69 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Acute Hypogonadism in Male Patients with Severe Traumatic Brain Injury Alexandre Hohl, Marcelo Fernando Ronsoni, Mariana Costa Silva, Maria Emilia Thais, Marisa Cesar Coral, Roger Walz University Hospital - Federal University of Santa Catarina, Florian[oacute]polis, Brazil; University Hospital - Federal University of Santa Catarina, Florian[oacute]polis, Brazil [bold]Objectives:[/bold] The hypothalamic-pituitary axis may change in traumatic brain injury (TBI), with subsequent hypopituitarism(1,2). Late gonadotrophin deficiency after TBI has been demonstrated frequently: 1-32% (3,4). The present study evaluates the total testosterone and calculated free testosterone in male patients with severe TBI in acute phase and its relationship with mortality.[br][bold]Methods:[/bold] We included 60 consecutive male patients with severe TBI (Glasgow Coma Scale score 8 or lower after acute neurosurgical resuscitation, or deterioration to that level within 48 hours of impact), admitted to the intensive care unit of the Hospital Governador Celso Ramos between April 2006 and April 2008. This is a public reference hospital for TBI covering a population of approximately 1 million, in the metropolitan area of Florianopolis, Brazil. Victims of gunshot injury and patients who evolved to brain death before 24 hours of admission were excluded. Total testosterone and SHBG (for calculation of free testosterone) were determined in blood plasma on the first, second, and third day after TBI (12, 30 and 70 hours after TBI, respectively). The study was approved by the ethics committee and the family responsible for the patient signed a consent form.[br][bold]Results:[/bold] Of 60 male patients evaluated, the average age of patients was 35 years. Outcome for 40 patients survived and 20 died, during hospitalization (33.3%). The causes of TBI were traffic accidents (60%), fall (28%), aggression (6%) and others (6%). The mean total testosterone (normal range [gt] 300 ng/dL) fell significantly from first to last evaluation: 438, 278 and 196 ng/dL, respectively. The same situation was observed with the calculated free testosterone (normal range [gt] 6.5 ng/dL): 12.3, 7.9 and 5.0 ng/dL, respectively. Both analyzed hormones showed a high prevalence of abnormalities ranging from 36.5% of patients with low total testosterone in first sample up to 83.3% of cases with low total and calculated free testosterone in the last sample. However, there was no statistically significant difference between the group of patients who survived and those who died during hospitalization (p[ge] 0.13).[br][bold]Conclusions:[/bold] Testosterone decreased progressively in the first 70 hours after severe TBI in male patients. A probable recovery occurs at a later period, since only a few of these men develop permanent hypogonadism. There was no relationship between levels of total and free testosterone and mortality among the patients studied.[br][br](1)Martins ET, Linhares MN, Sousa DS, Schroeder HK, Meinerz J, Rigo LA, et al. Mortality in severe traumatic brain injury: a multivariated analysis of 748 Brazilian patients from Florianopolis City. J Trauma. 2009;67(1):85-90. (2)Pinto HF, Weber TR, Rohr JF, Linhares RM, Coral MCH, Hohl A. Late evaluation of the somatotroph axis in survivors of severe traumatic brain injury. Arquivos Catarinenses de Medicina (Online). 2010;39(4):68-74. (3)Hohl A, Mazzuco TL, Coral MH, Schwarzbold M, Walz R. Hypogonadism after traumatic brain injury. Arq Bras Endocrinol Metabol. 2009;53(8):908-14. (4)Hohl A, Daltrozo JB, Pereira CG, Weber TR, Pinto HF, Gullo Jda S, et al. Late evaluation of the pituitary-gonadal axis in survivors of severe traumatic brain injury. Arq Bras Endocrinol Metabol. 2009;53(8):1012-9.[br][br]Sources of Research Support: Research supported by National Counsel of Technological and Scientific Development (CNPq) and Foundation for Research and Innovation of the State of Santa Catarina (FAPESC).[br][br]Nothing to Disclose: AH, MFR, MCS, MET, MCC, RW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1093 175 1126 SUN-69 PO36-02 Sunday 921 2012


920 ENDO12L_SUN-70 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Polymorphism in the Splicing Site of the Cryptic Exon of [italic]LHCGR:[/italic] Functional Consequences and Associations with Testosterone Level Jie Qiao, Wei Liu, Bing Han, Jia-Jun Wu, Bing-li Liu, Chun-Ming Pan, Shuang-Xia Zhao, Mind-Dao Chen, Huai-Dong Song Shanghai Ninth People[apos]s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China; Shanghai Institute of Endocrinology, Shanghai Jiao Tong University School of Medicine, Shanghai, China; Shanghai Jiao Tong University School of Medicine, Shanghai, China Luteinizing hormone/choriogonadotropin receptor (LHCGR) belongs to a subfamily of G protein-coupled receptors (GPCRs) that is responsible for transducing extracellular signals into intracellular responses by activating the G protein cascade. Selective splicing is proved to be a feature of this kind of glycoprotein receptor. Some [italic]LHCGR [/italic]splice variants were described in the human and other species, which was demonstrated to be caused by alternative splicing and exon skipping. A cryptic exon was recently identified which is derived from the potential splicing sites in the intron 6 resulting in intron retention and produce a cryptic exon----exon6A.[br]During the splicing functional research of a LCH patient with IVS6-3 C[rarr]A mutation, cryptic exon 6A was incidentally detected, including either 159 bp (short) or 207 bp (long) of the internal exon 6A. Owing to the existence of stop condon in the exon 6A, the translated product was the same truncated protein with only 209 amino acids. [italic]In vitro[/italic] transfection and Co-IP study revealed that the truncated receptor could neither bind the I[bold][sup]125[/sup][/bold]-hCG, nor give rise to dominant negative effect by dimerizing with wild-type receptor. Polymorphic variations were analyzed in 100 normal subjects and three SNPs were identifed. rs68073206 (48802225T/G) polymorphism was found to be located at position +5 in the splice donor site of intron 6A (following the long transcript), with predictable functional consequences. The allele frequency was 65.5 and 34.5% for wild type T allele and variant G allele, respectively. [italic]In vitro[/italic] mutagenesis and transfection study demonstrated the T[rarr]G variation would significantly increase the expression level of long transcript (almost 10 times), while the wild-type receptor transcript without 6A mildly increased (1.5 times). Moreover, in the human granulosa cells, real-time PCR revealed that long transcript expressed at very low even undetectable level in T/T genotype, mildly increased in the T/G genotype, but remarkably increased in the G/G genotype. G allele also elevated the expression level of wild-type receptor transcript. FSH, LH and testosterone level was also investigated in the 100 healthy male volunteers with three different genotypes.[br]Polymorphsm in the splicing site of the cryptic exon of LHCGR may impact on the expression proportion of different splicing variants, with fine adjustment of the hormone secretion.[br][br]1. Ascoli M, et a1. Endocr Rev. 2002.23: 141-174. 2. Minegishi T, et al. Molecular Human Reproduction 1997; 3:101-107. 3. Nakamura K, et al. Molecular Endocrinology 2004; 18: 1461-1470. 4. Kossack N, et al. PLoS Med 2008;5:e88.[br][br]Sources of Research Support: National Natural Science Foundation Program (81070666).[br][br]Nothing to Disclose: JQ, WL, BH, J-JW, B-lL, C-MP, S-XZ, M-DC, H-DS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 933 175 1127 SUN-70 PO36-02 Sunday 922 2012


921 ENDO12L_SUN-71 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Prevalence of Erectile Dysfunction Is Greater in Young Middle-Aged HIV-Infected Men Than in HIV-Uninfected Men Bruno Madeo, Daniele Santi, Stefano Zona, Giovanni Guaraldi, Antonio Granata, Cesare Carani, Vincenzo Rochira Unit [amp] Chair of Endocrinology [amp] Metabolism, University of Modena [amp] Reggio Emilia, Azienda USL of Modena, Modena, Italy; Metabolic Clinic, Infectious Diseases Unit, University of Modena [amp] Reggio Emilia, Modena, Italy Introduction. Erectile dysfunction (ED) is common among elderly men and patients suffering from chronic diseases, the latter probably including also HIV infection. No studies, however, compared the prevalence of ED in HIV-infected and -uninfected individuals using the IIEF-15.[br]Aim. To compare ED prevalence in young-middle aged men with and without HIV infection using the International Index of Erectile Function (IIEF-15) questionnaire.[br]Methods. We conducted a cross-sectional, observational, controlled study on 444 HIV-infected and 71 -uninfected men. The IIEF-15 questionnaire was used to assess ED. A cutoff score of [lt]25 of the erectile domain was used to diagnose ED. Serum testosterone, demographic and anthropometric (weight, height, BMI) characteristics were obtained from all participants. Statistics included the T-test, the Fisher[apos]s test, univariable and multivariable logistic regression and univariate and multivariate Spearman[apos]s correlation analysis.[br]Results. HIV-uninfected group was significantly younger than HIV-infected and presented a higher BMI (p[lt]0.001). The prevalence of mild, moderate, and severe ED was higher in HIV-infected than in HIV-uninfected men of all decades of age. In univariate analysis, HIV infection was associated with ED (OR=34.19, p[lt]0.001). In multivariable logistic regression analysis, HIV infection remained the strongest predictors of ED (OR=42.26, p[lt]0.001) followed by hypogonadism, after adjusting for age and BMI.[br]Conclusions. This study demonstrates a clear association between ED and HIV infection, after adjusting for age and BMI. Other than HIV infection, hypogonadism was associated with ED. In addition, the prevalence of ED was higher in HIV-infected than in HIV-uninfected men, in all decades of age. The early onset of ED in HIV-infected men could be considered a peculiar clinical hallmark of HIV and confirms precocious aging in these patients. ED should be of concern to clinicians when managing HIV-infected men even if the latter are young or middle aged.[br][br]Nothing to Disclose: BM, DS, SZ, GG, AG, CC, VR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 488 175 1128 SUN-71 PO36-02 Sunday 923 2012


922 ENDO12L_SUN-72 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Metabolic and Hormonal Evaluation in Patients with Erectile Dysfunction after Radiotherapy for Prostate Carcinoma Antonio Mancini, Luca Tagliaferri, Sebastiano Raimondo, Giovanna Mantini, Gian Carlo Mattiucci, Stefano Luzi, Alfredo Pontecorvi, Vincenzo Valentini, Numa Cellini Catholic University of the Sacred Heart, Rome, Italy; Catholic University of the Sacred Heart, Rome, Italy Erectile dysfunction (ED) is a frequent problem in patients treated for prostate carcinoma, profoundly affecting quality of life. It can recognize a multifactorial etiology, including ageing per se, metabolic, cardiovascular, endocrine, neurological and iatrogenic factors. Hormones, other than testosterone (T), influence sexual function; we previously demonstrated an increased estradiol concentration in patients affected by venous leakage ED.[br]In order to evaluate the endocrine component, we have evaluated a group of 19 patients, aged 58-76 ys., treated by radiotherapy and antiandrogen pharmacological therapy (bicalutamide), with ED, studying: metabolic parameters (glycemia, total HDL LDL cholesterol, triglycerides, uric acid, albumin), hormones (T, LH, FSH, estradiol, dihydrotestoterone, SHBG, IGF-1, PRL, FT3, FT4, TSH, insulin), evaluation of international index of erectile failure (IIEF); main vascular and neurological factors were excluded on the basis of basal Doppler evaluation and vibration threshold.[br]5 patients, previously treated with LHRH analogues, were still hypotestosteronemic (mean [plusmn] SD 1.03 [plusmn] 0.5 ng/ml), while in the other patients testosterone exhibited normal values (6.3 [plusmn] 3.04 ng/ml). Testosterone/estradiol molar ratio markedly different in the two groups (0.60 [plusmn] 0.23 in hypo-T patients and 1.60 [plusmn] 0.28 in normo-T patients); IIEF were significantly lower in hypo-T than in normo-T patients (4.2 [plusmn] 7.4 vs 10.4 [plusmn] 10.8). Metabolic parameters were markedy worse in hypo-T vs normo-T (HOMA index 6.7 [plusmn] 0.8 vs 2.4 [plusmn] 0.2).[br]These preliminary data suggest that, despite similar radiotherapic schedule treatment, sexual function is strictly related to hormonal milieu and metabolic status. A systematic approach to evaluate ED is mandatory in such patients, since improving some components ED can be positively influenced by personalized therapy.[br][br]Nothing to Disclose: AM, LT, SR, GM, GCM, SL, AP, VV, NC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 701 175 1129 SUN-72 PO36-02 Sunday 924 2012


923 ENDO12L_SUN-73 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Testocreme[reg], a Unique High-Potency Transdermal Hormone Cream Abraham Harvey Kryger Monterey Preventive Medical Clinic, Monterey, CA Testocreme[reg] is a new transdermal system containing 2.5, 5 or 10 percent testosterone allowing a small volume of cream to be applied as supplemental therapy. The main cause for concern when using any testosterone product is the possible interpersonal transfer of testosterone. This reduces the chance of skin-to-skin transference to a loved one. Testocreme can be applied to scrotal skin, where it preferentially converts to dihydrotestosterone due to a patented aromatase inhibitor. The scrotum is considered to have a fivefold higher resorption rate than other skin areas.[br]The primary objective of this three-year trial was to determine the levels of testosterone in hypogonadal men, using two dosages of Testocreme over long term. The secondary objectives of this trial were to evaluate pharmacokinetic and endocrine parameters after daily transdermal administration of Testocreme. This was an open label, randomized, parallel-group trial in symptomatic hypogonadal men with deficient free testosterone. Men (n=458) ranging from 35 to 75 years of age with a BMI of less than 32 kg/m2, were selected as subjects and compared to controls.[br]The main goal of testosterone supplementation is to restore total testosterone levels to physiological levels. In the trials, two different volumes of Testocreme: [frac12] and 1 gram, compared serum testosterone and sex hormone binding levels and the pre-post changes of the areas under the curve of testosterone and free testosterone, obtained at 30, 90 and 120 days, 12 hours after application for steady state levels. With respect to safety, routine laboratory parameters including liver function, hematocrit and hemoglobin were monitored and levels of prostate-specific antigen were measured over three years without significant change.[br]Statistically significant results, determined by Esoterix Labs demonstrated that Testocreme is both efficacious and consistent in the maintenance of stable therapeutic levels. Testocreme has overcome many of the disadvantages of other forms of testosterone replacement; creating a more physiologic testosterone level. Skin reactions are minimal and scrotal application is both convenient and discrete. With significant advantages in efficacy, transference issues, safety and cost suggest Testocreme as a favorable treatment for hypogonadal patients. Testocreme also improves erectile function without the need for erection enhancers and increases penis head sensitivity while optimizing hormonal levels at low volumes.[br][br]1. Araujo AB, O[apos]Donnell A, Brambilla DJ, et al. Prevalence and incidence of androgen deficiency in middle-aged and older men: estimates from the Massachusetts male aging study. J Clin Endocrinol Metab. 2004;89:5920-5926. 2. Araujo AB, Esche GR, Kupelian V, et al. Prevalence of symptomatic androgen deficiency in men. J Clin Endocrinol Metab. 2007;92:4241-4247. 3. Feldman HA, Longcope C, Derby CA, et al. Age trends in the level of serum testosterone and other hormones in middle-aged men: longitudinal results from the Massachusetts male aging study. J Clin Endocrinol Metab. 2002;87:589-598. 4. Bhasin S, Cunningham GR, Hayes FJ, et al. Testosterone therapy in adult men with androgen deficiency syndromes: an endocrine society clinical practice guideline. J Clin Endocrinol Metab. 2006;91:1995-2010. 5. ClinicalTrials.gov. A Study of the Efficacy of Using Testosterone in Older Men. http://www.clinicaltrials.gov identifier NCT00799617. Accessed September 28, 2009. 6. Buvat J, Lemaire A. Endocrine screening in 1,022 men with erectile dysfunction: clinical significance and cost-effective strategy. J Urol. 1997;158:1764-1767. 7. Kelleher S, Conway AJ, Handelsman DJ. Blood testosterone threshold for androgen deficiency symptoms. J Clin Endocrinol Metab. 2004;89:3813-3817. 8. Wang C, Nieschlag E, Swerdloff R, et al. Investigation, treatment and monitoring of late-onset hypogonadism in males: ISA, ISSAM, EAU, EAA and ASA recommendations. Eur J Endocrinol. 2008;159:507-514. 9. [uuml]hnert B, et al. Testosterone substitution with a new transdermal, hydroalcoholic gel applied to scrotal or non-scrotal skin: a multicentre trial. Euro J of Endo 2005;153:317-326. 10. Rolf C, et al. Interpersonal testosterone transfer after topical application of a newly developed testosterone gel preparation. Clin Endocrinol 2002;56:637-41. 11. Mohr B, Guay AT,O[apos]Donnell AmyB, McKinlay JB. Normal, bound and nonbound testosterone levels in normally ageing men: results from the Massachusetts Male Ageing Study. Clinical Endocrinol (2005) 62, 64[ndash]73Blackwell Publishing, Ltd.[br][br]Nothing to Disclose: AHK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1061 175 1130 SUN-73 PO36-02 Sunday 925 2012


924 ENDO12L_SUN-74 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Effects of Baseline Testosterone Levels on Symptom Improvement in Hypogonadal Men Receiving Testosterone Replacement Therapy Xiao Ni, David Muram Eli Lilly and Company, Indianapolis, IN; Eli Lilly and Company, Indianapolis, IN It is possible that symptom improvement may be affected by baseline testosterone (T) level in men receiving testosterone replacement therapy (TRT). We performed a post-hoc analysis to evaluate the effects of baseline T level on changes in sexual function and mood in hypogonadal men receiving 2% testosterone in a topical solution applied daily to the axillae.[br]An open-label trial (1) was conducted in 155 androgen-deficient men started on daily 60 mg T. Dose was adjusted on days 45 and 90 based on average serum T on days 15 and 60, respectively, as necessary to maintain T in the physiologic range (300-1050ng/dL). Sexual desire, sexual activity, positive mood, negative mood, percent full erection and erection maintained were assessed by the Psychosexual Daily Questionnaire (PDQ) (2) for 7 days preceding visits at days 1, 15, 60 and 120. Subjects were divided into four subgroups depending on baseline (pre-day 1) total serum testosterone (TT)[lt]100, 100[lt]200, 200[lt]300, and [ge]300 ng/dL. For each PDQ parameter, change from baseline to days 15, 60 and 120 was analyzed using repeated measures analysis of covariance (ANCOVA) adjusted for TT group, age, body mass index (BMI) and visit, with baseline TT subgroup effect assessed by the type III test of fixed effects.[br]All PDQ domains showed significant (p[lt]0.05) improvement for subjects in all TT subgroups. In general, improvement was maintained or increased during the 120-day treatment. When subgroups were compared, T levels at baseline did not appear to affect the magnitude of improvement; p[gt]0.05 for all PDQ domains. Subjects in the lowest TT group (TT[lt]100ng/dL) seemed to have the greatest numerical improvement in sexual desire, sexual activity, percent full erection, and erection maintained after adjusting for baseline age, BMI and visit. For example, the least squares (LS) means of the TT groups (main effects, averaged across other factors) for sexual desire were 1.70, 1.15, 1.13 and 0.97 for the TT[lt]100, [lt]200, 200[lt]300, and [ge]300ng/dL groups respectively. This trend was not observed in the positive and negative mood domains.[br]The study is limited by lack of a placebo control group and a relatively small number of patients with very low serum T levels at baseline. While adjustments were made for age and BMI, other confounders may have affected patients[apos] response to therapy. Additional research is needed to better understand the effects of TRT on symptom improvement in men with hypogonadism.[br][br](1) Wang C et al., J Clin Endocrinol Metab 1996;81:3578. (2) Lee KK et al., J Androl 2003;24:688.[br][br]Sources of Research Support: Eli Lilly and Company.[br][br]Disclosures: XN: Researcher, Eli Lilly & Company. DM: Clinical Researcher, Eli Lilly & Company. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 562 175 1131 SUN-74 PO36-02 Sunday 926 2012


925 ENDO12L_SUN-75 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Improvement in Sexual Function and Mood as a Function of Testosterone Level in Hypogonadal Men Receiving Testosterone Replacement Therapy Xiao Ni, David Muram Eli Lilly and Company, Indianapolis, IN; Eli Lilly and Company, Indianapolis, IN Based on limited available research (1-3) we undertook this post-hoc analysis to gain insights into symptomatic improvement in hypogonadal men on testosterone replacement therapy (TRT) based on whether they reached the pre-specified threshold of serum testosterone (T) 300 ng/dL.[br]This open-label trial (4) enrolled 155 testosterone-deficient (T[lt]300ng/dL) men given an initial daily dose of T 60 mg in a topical solution applied to the axillae. Dose was adjusted on days 45 and 90 when necessary to maintain T within the physiological range (300-1050ng/dL). Sexual function and mood changes were assessed for 7 days preceding visits on days 1, 15, 60, and 120 by the Psychosexual Daily Questionnaire (PDQ) (5). Subjects were divided into two groups (T[lt]300 and [ge]300 ng/dL) based on their T level during therapy. Analysis of covariance (ANCOVA) with adjustments for baseline PDQ scores, age, and body mass index (BMI) was used to evaluate change in PDQ scores from baseline (pre-day1) to pre-day15 and pre-day 120 (with last-observation-carried-forward for dropouts) both within and between the groups.[br]As early as day 15, numerical improvement was observed in all PDQ scores (sexual desire, sexual activity, percent full erection, erection maintained, and positive and negative mood) for both groups. Within-group improvement was significant (p[lt]0.05) for all parameters except positive mood in group T[lt]300. Improvement in PDQ scores was maintained or increased on day 120. For example, sexual desire changes on day 15 for the T[lt]300 and [ge]300 ng/dL groups were 1.1 (0.2) and 0.8 (0.1) (least square means [standard error]) respectively; on day 120 values were 1.1 (0.3) and 1.5 (0.1) respectively. On day 120, changes in PDQ scores were numerically greater in the T[ge]300 ng/dL group than in the T[lt]300 ng/dL group, although the between-group difference was not significant (p[gt]0.05).[br]These data show that symptoms in hypogonadal men receiving TRT improved by day 15 and continued until day 120. Improvements were noted even in those with T[lt]300 after treatment. The study is limited by lack of a placebo control group and a small number of patients with serum T levels [lt]300ng/dL after treatment and also that the analysis groupings (T groups) are based on post-baseline efficacy results. Additional research is needed to better understand the thresholds at which patients and physicians can expect symptoms of hypogonadism to improve.[br][br](1) Kelleher S et al., J Clin Endocrinol Metab 2004;89:3813. (2) Zitzmann M et al., J Clin Endocrinol Metab 2006;91:4335. (3) Saad F et al., Eur J Endocrinol 2011;165:675. (4) Wang C etal., J Clin Endocrinol Metab 1996;81:3578. (5) Lee KK et al., J Androl 2003;24:688.[br][br]Sources of Research Support: Eli Lilly and Company.[br][br]Disclosures: XN: Researcher, Eli Lilly & Company. DM: Clinical Researcher, Eli Lilly & Company. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 574 175 1132 SUN-75 PO36-02 Sunday 927 2012


926 ENDO12L_SUN-76 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Elevation of Testosterone Concentrations in the Low Normal Range Reduces Fatigue in Older Men Elizabeth Hoang, William Durham, Shanon Casperson, Charles Gilkison, Lichar Dillon, Melinda Sheffield-Moore, Randall Urban University of Texas Medical Branch, Galveston, TX Low concentrations of testosterone are associated with increased perception of fatigue. While testosterone supplementation may increase skeletal muscle mass, little is known about how factors such as mode and duration of administration, pre- and post-treatment testosterone concentrations, and changes in muscle mass will affect perceptual fatigue. In this study, we hypothesized that 1) testosterone treatment in men with low normal serum testosterone would reduce perceptual fatigue even prior to significant accrual of muscle mass, and 2) short-term (1 week) changes in perceptual fatigue responses would not be dependent on mode of administration (gel vs. injection). Nine healthy older men (age: 60-85 yrs, testosterone: [le]500 ng/dL) were studied during a baseline week and a treatment week during which they received either a testosterone injection (single 100 mg dose) or daily testosterone gel application (10 mg/day). The Brief Fatigue Inventory, a short survey assessing fatigue, was completed daily throughout the two-week study and analyzed to determine the subjects[apos] responses to testosterone supplementation. Comparisons were made between baseline and treatment periods as well as between gel and injection treatment groups. In addition, daily activity levels were measured using accelerometers (currently being analyzed). These preliminary results suggest that testosterone treatment reduces fatigue, and that short-term responses in fatigue may be dependent on administration mode. Future directions should include longer-term interventions to determine whether testosterone and fatigue responses become less dependent on administration mode with time, and to determine if testosterone-induced changes in muscle mass and function further reduce fatigue.[br][br]Nothing to Disclose: EH, WD, SC, CG, LD, MS-M, RU 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 416 175 1133 SUN-76 PO36-02 Sunday 928 2012


927 ENDO12L_SUN-77 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) 47,XXY Klinefelter Syndrome Associated with Partial Androgen Insensitivity Phenotype Conferred by a Complete Inactivating Mutation in the Androgen Receptor Gene Luciane Carneiro de Carvalho, Alina Coutinho Rodrigues Feitosa, Andresa De Santi Rodrigues, Mirian Yumie Nishi, Rosana Barbosa Silva, Elaine Maria Frade Costa, Sorahia Domenice, Berenice Bilharinho Mendonca HC-FMUSP, S[atilde]o Paulo, Brazil; Escola de Medicina e Sa[uacute]de P[uacute]blica da Bahia, Bahia, Brazil [bold]Background:[/bold] Klinefelter syndrome (KS) and androgen insensitivity syndrome are among the most common disorders of sex development and rarely both syndromes can affect the same individual (1). [bold]Clinical case[/bold]: A 19 years old healthy woman sought medical attention due to primary amenorrhea and severe pain in both inguinal regions. At physical examination she was a proportional tall overweighed young female (height=173.3 cm (SD +1.85); weight: 89.4 kg (SD +4.2), BMI: 30; Breast development was Tanner V and pubic hair was normal at Tanner IV stage. External genitalia examination disclosed an enlarged clitoris (3.5 x1.5 cm), two perineal opening and bilateral inguinal gonads. Her karyotype was 47,XXY (G banding) and ultrasonography of the inguinal region disclosed a 3.6 mL structure on the right and a 4.9 mL structure on the left, compatible with male gonads. Hormonal investigation revealed elevated basal LH (20 U/L - n: 0.4 to 5.7) and FSH (35 U/L -n: 1.1 to 13.5) levels and testosterone levels at normal male levels: (325 ng/dL- n: 260 to 990). She has undergone bilateral orchiectomy at age 20, and anatomopathological result was consistent with atrophy of seminiferous tubules. Klinefelter syndrome is rarely associated with ambiguous genitalia, mainly with almost female external genitalia. This fact associated with high LH levels and normal male testosterone levels and normal breast development suggested androgen insensitivity syndrome. The analysis of the androgen receptor ([italic]AR[/italic]) gene, using genomic DNA revealed a heterozygous insertion of an adenosine at nucleotide 3070 (AAT[gt] AAAT) in exon 7, resulting in an exchange of asparagine for lysine at position 849 protein, generating a frameshift which leads to a stop codon 30 aminoacids ahead (p.Asn849Lysfs*30). This mutation has been previously described in several patients with CAIS (2). The PAIS phenotype of our patient suggests that normal [italic]AR[/italic] is partially expressed. The heterozygous state of [italic]AR[/italic] mutation points to a maternal (Xm1Xm2Y) or paternal (XmXpY) nondisjunction event in meiosis I, but parents[apos] DNA were not available to confirm this hypothesis. [bold]Conclusion[/bold] We report a female patient with KS and [italic]AR[/italic] mutation that would generate a CAIS phenotype, but resulted in a PAIS phenotype due to partially expression of normal [italic]AR[/italic]. We emphasize the importance of considering this association in KS patients with ambiguous genitalia.[br][br](1) Uehara S, et al., Am J Med Genet 1999; 86:107-11. (2) Ahmed SF, et al., J Clin Endocrinol and Metabol 2000; 85: 658-665.[br][br]Nothing to Disclose: LCdC, ACRF, ADSR, MYN, RBS, EMFC, SD, BBM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1113 175 1134 SUN-77 PO36-02 Sunday 929 2012


928 ENDO12L_SUN-78 POSTER SESSION: Male Reproductive Endocrinology Case Reports (1:30 PM-3:30 PM) Estrogen Deficiency in a Male with a Congenital Aromatase Gene Mutation Suzan Saber, Susan V Bukata, Stephen R Hammes University of Rochester Medical Center, Rochester, NY; University of Rochester Medical Center, Rochester, NY Background[br]Aromatase gene mutations in men have been reported only four times. Therefore, diagnosis can be delayed and appropriate estrogen replacement strategies are not well established.[br]Case[br]A 25-year old male who sustained a fracture of the left fifth proximal phalynx was found to have markedly delayed bone maturation and open growth plates throughout the hand and wrist. He reported forearm and phalynx fractures while playing sports in the past. He was the tallest in his family at 5[apos]11[quot]. He had normal secondary sexual characteristics and normal puberty at age 13, with continued linear growth since then. Exam revealed minimal genu valgum, normal genitalia and testicular volume. He had mild acanthosis under his arms. His bone mineral density revealed a T-score of -3.5 in the left femoral neck and -3.5 in the L-spine. His total testosterone=782 ng/dl (240-950 ng/dl), DHT=523 pg/ml (106-719 pg/ml), and estradiol[lt]5 pg/ml (8-43 pg/ml). Fasting lipids, glucose, insulin, and glucose tolerance test were normal.[br]Aromatase deficiency was suspected. Gene testing revealed a homozygous point mutation at the end of exon V. This G to A mutation leads to aberrant mRNA splicing, resulting in a truncated, inactive aromatase. Interestingly, the patient[apos]s older sister had a history of amenorrhea requiring estrogen replacement. Genetic testing following our patient[apos]s results revealed the same mutation in her. Further discussion revealed that his parents were consanguineous. On literature review, two case reports of a woman and a man describe the same mutation, although their physical findings were slightly more severe than in our patient. The patient was started on 25 mcg estradiol patch once weekly with normalization of his estradiol to 35 pg/ml. Three months after starting the patch, the patient reported hand stiffness and knee pain just distal to the patella. He also developed nipple tenderness but no gynecomastia. His libido remained normal. At eight months his hand film showed minimal interval maturation, but all hand growth plates were still open. His bone density demonstrated increases of 8.2% and 5.9% at the spine and hips, respectively.[br]Conclusion[br]Aromatase deficiency in males results in phenotypes of variable severity. Therefore, this diagnosis should be considered in any man with delayed epiphyseal closure. Estrogen treatment involves striking a balance between the need for skeletal maturation and the maintenance of male sexual characteristics.[br][br]Nothing to Disclose: SS, SVB, SRH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 857 176 1135 SUN-78 PO51-01 Sunday 930 2012


929 ENDO12L_SUN-79 POSTER SESSION: Male Reproductive Endocrinology Case Reports (1:30 PM-3:30 PM) Elevated Serum Testosterone and Sex Hormone Binding Globulin Associated with Sexual Dysfunction: A Familial Disorder? Thanh Duc Hoang, Vinh Q Mai, Patrick W Clyde, KM Mohamed Shakir Walter Reed National Military Medical Center, Bethesda, MD Background: Elevated sex hormone binding globulin (SHBG) levels have been reported to be associated with increased insulin sensitivity, hyperthyroidism, reduced risk of type 2 diabetes mellitus (1,2) and sexual dysfunction in men with chronic hepatitis C infection (3). We report a familial case of hypertestosteronemia and elevated SHBG associated with decreased libido and erectile dysfunction.[br]Clinical case: A 37 y/o male with a 2-year history of gradually decreased libido and erectile dysfunction was found to have repeated elevation of serum testosterone and SHBG levels. He shaves daily and denies any headaches, vision changes, breast enlargement, chest pain, shortness of breath, or weight changes. Past medical history was significant for shingles and seasonal allergy. He was taking a multivitamin daily. He never smoked and drank 2 beers weekly. He has 2 healthy children. He reports a family history of hypertension and diabetes mellitus. Vital signs: BP 122/71, HR 60 bpm, weight 176 lbs, height 72 in, and BMI 24 kg/m[sup]2[/sup]. Physical examination: normal thyroid, no gynecomastia, no galactorrhea, normal phallus, testicles 25 cc bilaterally without palpable mass. Heart, lungs and neurological examination was normal. Lab results: 8 A.M serum total testosterone 1186-1448 ng/dL (nl 250-1100), free testosterone 141-169 pg/mL (nl 35-155), SHBG 86-98 nmol/L (nl 10-50), LH 8.8 mIU/mL (nl 1.5-9.3), FSH 6.0 mIU/mL (nl 1.4-18.1), estradiol 58.8 pg/mL (nl 7.6-42.6), and TSH 1.09 mcIU/mL (nl 0.27-4.20). Fasting glucose, prolactin, HCG, CBG, liver associated enzymes, hepatitis and congenital adrenal hyperplasia panels were normal. Clomiphene challenge test was normal. Abdominal CT scan showed normal liver and adrenal glands. Pituitary MRI, testicular ultrasound, baseline DXA scan, and EKG were all normal.[br]Testing of his 11 y/o daughter and 8 y/o son revealed SHBG values of 158 nmol/L (nl 24-120) and 120 nmol/L (nl 32-158) with total testosterone levels of 13 ng/dL and 5ng/dL (nl [lt]25), respectively. Estradiol and TSH levels are normal for both children.[br]Conclusions: The patient most likely has familial elevated SHBG leading to hypertestosteronemia, given the fact that his daughter also has elevated SHBG and his son with high-normal SHBG. To our knowledge, hypertestosteronemia due to familial elevated SHBG has not been reported previously. Patients with hypertestosteronemia and elevated SHBG need further investigations, including possible genetic studies.[br][br](1) Lakshman KM et al., J Gerontol A Biol Sci Med Sci 2010; 65:503-9. (2) Perry JR et al., Human Molecular Genetics 2010; 19: 535[ndash]544. (3) Rao J et al., J Clin Gastroenterology 2009; 43:94-95.[br][br]Nothing to Disclose: TDH, VQM, PWC, KMMS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 661 176 1136 SUN-79 PO51-01 Sunday 931 2012


930 ENDO12L_SUN-80 POSTER SESSION: Male Reproductive Endocrinology Case Reports (1:30 PM-3:30 PM) Sertoli Cell Hyperfunction with Low FSH in a Pubertal Boy with Bilateral Macroorchidism Romina Paula Grinspon, Paula Scaglia, Debora Braslavsky, Stella Campo, Ignacio Bergada, Horacio Domene, Mirta Gryngarten, Rodolfo Alberto Rey Hospital de Ni[ntilde]os Ricardo Guti[eacute]rrez, Buenos Aires, Argentina; Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina [bold]Background:[/bold] Bilateral macroorchidism has been described with no testicular hyperfunction in Fragile X syndrome, testicular adrenal rest tumors or other infiltrative disorders, and with testicular hyperfunction, due to increased gonadotropin signaling activity, in gonadotropin-secreting adenomas, McCune-Albright syndrome, aromatase deficiency and severe hypothyroidism. Testis size is mainly dependent on Sertoli cell number before puberty and on germ and Sertoli cell number after pubertal onset. FSH is the most conspicuous regulator of Sertoli cell proliferation, by activating FSH-R coupled to Gs[alpha] protein.[br][bold]Clinical case:[/bold] A 10-yr-old boy presented with a remarkable discordance between enlarged testes (20 ml) and penis and pubic hair corresponding only to incipient Tanner stage 3. He had bilateral mild gynecomastia. No signs of Fragile X or McCune-Albright syndrome, infiltrative disorders, CAH, aromatase deficiency or hypothyroidism were present. Inhibin B was abnormally high at 464 pg/ml (Tanner 3 range: 126-257), and basal FSH was low at 0.76 IU/L (1.43-7.44) with no response to GnRH (0.97 IU/L). Discordantly, T was 263 ng/dl (12-368) and basal LH 2.23 IU/L (0.67-4.65) with a normal response to GnRH (15.8 IU/L). E2 was 14 pg/mL (10-35). During 2-yr follow-up, testicular size increased to [gt]25 ml and penis and pubic hair progressed to Tanner stage 5. Gynecomastia regressed. Inhibin B persisted high at 628 pg/ml (118-340) with FSH low at 0.82 (1.14-6.99) and normal T and LH. With suspicion of FSH-R constitutive activation,[italic] FSHR[/italic] gene coding and flanking intronic regions (exons 1 to 10) were sequenced: 3 SNPs in heterozygosis in noncoding regions and 2 SNPs in homozygosis in coding regions were identified, but no allegedly activating mutation was found.[br][bold]Conclusion: [/bold]Macroorchidism, with normal pituitary-gonadal axis function, may be a normal variant. Alternatively, macroorchidism may be a sign of gonadal hyperfunction. Increased testicular activity may involve both tubular and interstitial compartments or be dissociated. A dissociated primary testicular hyperfunction restricted to Sertoli cells is likely the underlying cause of macroorchidism in this patient. An activating mutation of the FSH-R could not be evidenced.[br][br]Sources of Research Support: Partially supported with PIP grant 11220090100615 (CONICET) to SC and RR and PICT grant 2008-0521 (ANPCYT) to RR.[br][br]Nothing to Disclose: RPG, PS, DB, SC, IB, HD, MG, RAR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1528 176 1137 SUN-80 PO51-01 Sunday 932 2012


931 ENDO12L_SUN-81 POSTER SESSION: Male Reproductive Endocrinology Case Reports (1:30 PM-3:30 PM) Klinefelter Syndrome and Testicular Microlithiasis [mdash] Case Report Selma Barbosa Souto, Daniel Carvalho Braga, Davide Mauriio Carvalho Centro Hospitar S[atilde]o Jo[atilde]o, Porto, Portugal; University of Porto, Porto, Portugal Introduction: Klinefelter[apos]s syndrome (KS) is characterized by small firm testes, gynaecomastia and hypogonadism. The classic karyotype is 47,XXY. It is expected that KS is underdiagnosed, with less than 10% of the diagnoses made before puberty. Testicular microlithiasis (TM) can be observed in association with several conditions, such as KS. It provides about 10-fold increased risk of testicular germ cell tumors or intratubular germ cell neoplasia of unclassified type in patients with features of testicular dysgenesis syndrome, such as KS. Case report: A 20 years-old patient was referred to endocrinology department due to bilateral painless bilateral gynaecomastia since 14 years old. No relevant personal or family history. From the physical examination, an obvious bilateral gynaecomastia was detected. He also had small firm testes ([lt]10 mL) with normal penis size and sparse facial hair. Olfaction and pubic hair were normal. No mental retardation, anemia appearance, varicosity, reduced muscle strength or malformation was noted. He was 182 cm and 62.8 kg, with an arm span of 178 cm and a ratio SS/SI (82/100) [lt] 1. His family target height was 169.5 cm. The blood tests revealed: FSH [43.84mUI/mL(0.0[ndash]18.0)] and LH [31.72mUI/mL (0.0[ndash]11.0), total testosterone [2.97ng/mL (2.8-8.0)], oestradiol [15.49 pg/mL(7.6-42.6)] and SHBG [39.60 mmol/L(10.0-50.0)]. Tests size measured by ultrasound was 21[times]18[times]10 mm (right test) and 22[times]17[times]10 mm (left test). The ultrasound also showed bilateral TM. Serum tumor markers, alfa fetoprote[iacute]na and beta-human chorionic gonadotropin were normal. Because of his gonadal dysfunction and skeletal proportions a KS was suspected. Cytogenetic studies of peripheral blood lymphocytes revealed a 47,XXY karyotype, and the diagnosis of KS was confirmed. Mammary plastic surgery to correct gynaecomastia was proposed to the patient who has refused. He started testosterone replacement therapy with increased muscle strength and facial hair. Discussion: Despite being considered a benign genetic disorder, patients with KS are predisposed to breast cancer, hematologic malignancies, and extragonadal midline germ-cell tumors. In the addition to KS, our patient revealed TM, increasing the risk of testicular tumors with a negative impact in fertility.[br][br]Nothing to Disclose: SBS, DCB, DMC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1292 176 1138 SUN-81 PO51-01 Sunday 933 2012


932 ENDO12L_SUN-82 POSTER SESSION: Male Reproductive Endocrinology Case Reports (1:30 PM-3:30 PM) Birth of a Healthy Boy from a Patient with Nonmosaic Klinefelter Syndrome after IVF-ICSI with Preimplantation Genetic Diagnostics George Mskhalaya, Elena Zakharova, Mikhail Potapov, Victoria Zaletova, Evgeniy Efremov, Daria Gusakova Center for Reproductive Medicine MAMA, Moscow, Russian Federation; Center for Reproductive Medicine MAMA, Moscow, Russian Federation; Center for Reproductive Medicine MAMA, Moscow, Russian Federation; Russian Research Center for Urology, Moscow, Russian Federation Klinefelter[apos]s syndrome is a manifestation of one the most common sex chromosome aneuploidies in which an extra X-chromosome (80%), X-chromosome polysomy or mosaic forms are found in the male karyotype. The genetic imbalance is clinically manifested by primary hypogonadism and infertility. Histologically, degeneration of the germinal epithelium and hyalinosis of seminiferous tubules are observed. In a number of cases, patients achieved pregnancy using the IVF technique, however a high risk of sex chromosome aneuploidy in embryos is observed, which may result in birth of a sick child.[br]The spouses [italic]B.[/italic] and [italic]Ch[/italic]., 25 y.o., came to the Center with absence of pregnancy complaints. A cytogenetic analysis showed that the [italic]Patient B[/italic]. had nonmosaic Klinefelter[apos]s syndrome (47, XXY), an ejaculate analysis showed oligoasthenoteratozoospermia. The spouses were recommended to achieve pregnancy by the IVF-ICSI technique with preimplantation genetic diagnostics (PGD).[br]In the IVF-ICSI cycle, three embryos were obtained from the [italic]Patient Ch.[/italic] The embryos underwent a trophectoderm biopsy with a subsequent genetic analysis of numeric abnormalities in sex chromosomes and chromosome 21. The genetic analysis was performed by the FISH method using DNA probes CEP X, CEP Y, LSI 21 (Vysis, USA). The embryos had the following genetic status of the sex chromosomes: XX, XY, X0 monosomy. No aneuploidy of chromosome 21 was found. Male and female embryos were transplanted to the [italic]Patient Ch.[apos][/italic]s uterus, and a singleton pregnancy developed. On June 5, 2011, at gestation weeks 36-37, a live mature boy was born (3,100 g, 49 cm, Apgar score - 8-9). According to the genetic analysis results, the child[apos]s karyotype was 46, XY.[br]The above mentioned clinical case of pregnancy achieved by the IVF-ICSI (with PGD) and subsequent birth of a healthy boy from a patient with nonmosaic Klinefelter[apos]s syndrome is the first case in our country described in the literature.[br][br]Nothing to Disclose: GM, EZ, MP, VZ, EE, DG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 247 176 1139 SUN-82 PO51-01 Sunday 934 2012


933 ENDO12L_SUN-83 POSTER SESSION: Male Reproductive Endocrinology Case Reports (1:30 PM-3:30 PM) A Same Homozygous Mutation (R103X) in Exon 2 of 5-[alpha]-Reductase Type 2 Gene (SRD5A2) Presenting with Different Female External Genital Phenotype Enver Simsek, Cigdem Binay, Serdar Ceylaner Eskisehir Osmangazi University School of Medicine, Eskisehir, Turkey; Intergen Laboratory, Ankara, Turkey Background: Deletions and mutations of 5-alpha-reductase type 2 gene (SRD5A2) have been identified in 46,XY disorders of sexual differentiation.[br]Clinical case: A 18-month-old girl referred to endocrine department for investigation the etiology of 46,XY disorder of sexual differentiation (DSD). Family history revealed that her parents were first degree consanguineous and she had a four-year-old healthy female sibling. Physical examination revealed that the length was 73 cm (Standard Deviation Score [SDS], -2.3), weight 7.5 kg (SDS, -3.7) and head circumference 45 cm (SDS, 2.6). Genital examination revealed external female genitalia phenotype (Fig. 1A), no genital skin pigmentation and labioscrotal fusion, urethral opening was at the base of hypoplastic fallus, normal distal vaginal opening (Fig. 1B), and bilateral palpable mass (0.5 x 0.3 cm) at the distal ends of inguinal channels. Abdomino-pelvic ultrasound examination revealed that there was no mullerian structures and bilateral masses (5 x 4 mm) at the distal-end of inguinal channels. Hormonal analysis revealed that follicle stimulant hormone was 0.59 mIU/ml; luteinizing hormone, 0.19 mIU/ml; total testosterone, [lt]20 ng/ml; morning (800 h) cortisol, 6.08 mcg/dl (normal [gt]5.0 mcg/dl), 17-hydroxy progesterone, 0.12 ng/ml (normal 0.03-0.9 ng/ml), and dehydroepiandrosteron sulfate, [lt] 15 mcg/dl. Basal testosterone (T), dihydrotestosterone (DHT), androstenedione (A), and T/DHT ratio were [lt]20 ng/ml, 13.45 pg/ml (N:30 pg/ml), 0.1 ng/ml (N:0.08-0.5 ng/ml), and 14.8 (N: 11.1 +/- 4), respectively. After three dose injection of hCG, T, DHT, A, and T/DHT ratio were 142 ng/dl, 18.07 pg/ml, 0.4 ng/ml, and 78 (N[lt]20), respectively. Analysis of clinical and hormonal findings was consistent with the diagnosis of 5 alpha-reductase type 2 deficiency. Analysis consisted of PCR amplification and direct sequencing of the entire coding region of the SRD5A2 gene including intron-exon boundaries. This study revealed that the presented case has homozygous R103X mutation in SRD5A2 gene (Fig.2). This is a stop codon mutation and CGA was converted to TGA in codon 103. This is the second reported case of 5-alpha reductase deficiency associated with R103X mutation. The first case presented with cliteromegaly and our case with normal female external genital pheonotype (1).[br]Conclusion: There is no phenotype and genotype relation in 5-alpha reductase deficiency patients.[br][br](1) Hiort O et al.,Int J Androl 2002;25:55.[br][br]Nothing to Disclose: ES, CB, SC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 36 176 1140 SUN-83 PO51-01 Sunday 935 2012


934 ENDO12L_SUN-84 POSTER SESSION: Male Reproductive Endocrinology Case Reports (1:30 PM-3:30 PM) Dysgenetic Male Pseudohermaphrodite: Impact on Gender Identity along with Clinical Features [amp] Management Mohammed NMN Ahmed, Abdulrahman NMN Al Nuaim King Faisal Specialist Hospital [amp] Research Center, Riyadh, Saudi Arabia Male Pseudohermaphrodite (MPH) is defined as an individual whose genital differentiation is not that expected of normal male, despite 46, XY [amp] presence of testes. Differential Dx include: defect in androgen action, defects in testicular activity such as 17-KS reductase deficiency, Leydig cell agenesis/hypoplasia, or testes regression syndrome.[br]Aim: To define diagnostic [amp] management considerations for dysgenetic (MPH) due to defect in testicular activity [amp] its impact on gender identity.[br]Case Summary: A 19-year medical student raised as a female sought medical advise for amenorrhea, [amp] acne. O/E: ambiguous external genitailia, bilateral inguinal gonads, clitoromegaly, blind-ending vagina, Karyotype:46, XY. Hormonal evaluation: Serum testosterone (T)1.79 nml/l(RR: male 9.9-27.8) Dihydrotestosterone (DHT)0.19nmo/l(RR: 0.1-0.8).Following 4 days HCG stimulation; T increased 4.14, [amp] DHT increased 0.4, Androstenedione:3.3 nmol/l;(RR: 1.0-12.2), DHEAS: 5.29 nmol/l(1.8-8.3),17-OH-Progesterone 2.0(RR: 0.3-3.3), LH: 30.7 IU/l(RR: 1.7-8.6), E2: 32 pmol/l(RR: male 28-156 [amp] female 46-774), FSH:48 IU/l (RR: 1.5-12.4),Urology evaluation under GA:3 cm phallus, Cystoscopy findings: long male-like urethra, Vaginoscopy: blind-end 1.5 cm vagina, Laproscopy: wolffian duct derivatives (spermatic cords, atrophic testes, epididymis [amp] Vas deferens) identified, as evidence for fetal testicular testosterone production/action. There was absence of uterus, tubes [amp] ovaries, attesting to AMH generation/action from fetal sertoli cells during gestation.[br]Above stated findings were not in keeping w/5 alpha-reductase deficiency [amp] 17-KS reductase deficiency. Possibilities for this male pseudohermaphrodite included Leydig cell agenesis/hypoplasia, or testes regression syndrome. The Dx of leydig cell agenesis is based on little or no increase in testosterone to HCG/LH, mullerian regression is complete because secretion of AMH by sertoli cells is not impaired. Pt. was seen by 3 mental health experts rendering the opinion that Pt. [amp] family are well-adjusted with established female identity, with female sexual [amp] social adjustment. Pt. did not wish to change sexual identity, [amp] wanted orchidectomy. Bilateral orchidectomy was done, [amp] pt. remains satisfied. Histology: Sertoli-cell only.[br]Conclusion: MPH with ambiguous external genetalia develop female gender identity [amp] should be raised as females, with gonadectomy performed because of high risk of gonadoblastoma in dygenetic undesended testes.[br][br]Nothing to Disclose: MNMNA, ANMNAN 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 448 176 1141 SUN-84 PO51-01 Sunday 936 2012


935 ENDO12L_SUN-85 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) POMC Neurons Are a Distinct Leptin-Responsive Population in the Nucleus of the Solitary Tract Alastair S Garfield, Oliver J Marston, Mark L Evans, Lora K Heisler University of Cambridge, Cambridge, UK; University of Cambridge, Cambridge, UK The central melanocortin system represents a fundamental component of the neurological network regulating energy balance and is defined by the antagonistic actions of two peptidergic neurotransmitters; appetite suppressing pro-opiomelanocortin (POMC) and appetite stimulating agouti-related peptide (AgRP). The arcuate nucleus of the hypothalamus (ARC) is the preponderant source of melanocotinergic afferents within the mammalian brain, however a second smaller neuroanatomical niche of POMC expressing neurons is located within the nucleus of the solitary tract (NTS). Using transgenic reporter lines, we investigated the significance of this population of neurons to the physiological control of energy balance by leptin. Neurochemical characterisation of POMC-EGFP cells revealed them to be a subpopulation distinct from other leptin regulated energy balance related NTS neurons inasmuch as they do not co-express preproglucagon/GLP-1, cholecystokinin, or tyrosine hydroxylase/catecholamine; nor do they co-express brain-derived neurotrophic factor, nesfatin, or cocaine-amphetamine regulated transcript. However, POMC-EGFP and the leptin receptor (LepRb)-dtTOMATO cells of the NTS exhibited approximately 100% co-expression with the transcription factor paired-like homeobox 2b (Phox2b), a population of neurons important in leptin[apos]s effects on energy balance. Concomitant transgenic labeling of POMC and LepRb using a double reporter line determined that 30% of POMC-EGFP NTS cells expressed LepRbs. The responsivity of these neurons to leptin was further substantiated by physiological and electrophysiological interrogation and demonstrated their significant activation upon anorectic concentrations of leptin. Furthermore, loss of leptin tone onto NTS POMC cells in o[italic]b/ob [/italic]mice engenders a significant increase in endogenous [italic]Pomc[/italic] mRNA. Here we characterize a distinct population of LepRb expressing and leptin regulated NTS neurons, those expressing POMC-EGFP, that are of physiological relevance to leptin modulated appetite.[br][br]Nothing to Disclose: ASG, OJM, MLE, LKH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1849 177 1142 SUN-85 PO27-01 Sunday 937 2012


936 ENDO12L_SUN-86 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Hypothyroidism Compromises Leptin Production and Hypothalamic Leptin Signaling in Mice Claudia Groba, Steffen Mayerl, Theo J Visser, Veerle Darras, Andreas Habenicht, Heike Heuer Leibniz Institute for Age Research/Fritz Lipmann Institute, Jena, Germany; Erasmus Medical Center, Rotterdam, Netherlands; Katholieke Universiteit, Leuven, Belgium; Friedrich Schiller University, Jena, Germany The impact of thyroid hormone (TH) on metabolism and energy expenditure is well established but the role of TH in regulating nutritional sensing particularly in the CNS is only poorly defined. Here, we analysed the consequences of hypothyroidism on leptin production as well as leptin sensing in mice. For this purpose, we studied TRH receptor 1 (Trhr1) knockout mice that exhibit congenital hypothyroidism due to a diminished TRH stimulation of pituitary thyrotrophs. We also included in our experiments athyroid Pax8 ko mice as well as wild type mice rendered hypothyroid by MMI treatment.[br]Hypothyroid mice exhibited a 50% reduction in white adipose tissue compared to euthyroid controls. Moreover, the animals showed a decrease in leptin transcript levels as well as strongly reduced leptin serum levels. The impact of TH on leptin expression was further substantiated by in vitro experiments. Murine 3T3-L1 adipocytes responded to T3 with a rise in leptin transcript levels suggesting that TH indeed may regulate leptin expression in a cell-autonomous manner.[br]The transcript levels of leptin receptor were altered under hypothyroid conditions as well. Hypothyroid animals showed an increase in LepR mRNA expression specifically in neurons of the hypothalamic arcuate nucleus, an important target of leptin action. Likewise, the transcript levels of SOCS3, a potent negative regulator of LepR signaling, were strongly decreased in these neurons. In order to eliminate any effects of the different circulating leptin levels in our analysis, we generated hypothyroid and leptin-deficient animals by crossing hypothyroid Trhr1 ko mice with the leptin-deficient ob mice and monitored their response to leptin substitution. In comparison to euthyroid ob animals, hypothyroid Trhr1/ob ko mice showed decreased body weight and appetite loss following three days of leptin treatment. Moreover, phospho-Stat3 immunoreactivity and SOCS3 mRNA levels in the arcuate nucleus were strongly reduced in Trhr1/ob ko mice compared to euthyroid controls. These data indicate alterations in the intracellular processing of the leptin signal under hypothyroid conditions and thereby unravel a novel mode of action by which TH affects energy metabolism.[br][br]Nothing to Disclose: CG, SM, TJV, VD, AH, HH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 435 177 1143 SUN-86 PO27-01 Sunday 938 2012


937 ENDO12L_SUN-87 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Microglial Inactivation during HFD Feeding Promotes Weight Gain Joshua P Thaler, Chun-Xia Yi, Stephan Guyenet, Bang Hwang, Miles Matsen, Hong Nguyen, Gregory J Morton, Brent E Wisse, Denis E Baskin, Matthias Tschop, Michael W Schwartz University of Washington, Seattle, WA; Helmholtz Zentrum M[uuml]nchen, Munich, Germany; Puget Sound Veterans Administration Hospital, Seattle, WA Hypothalamic inflammation occurs during consumption of high fat diets (HFD) and is hypothesized to promote diet-induced obesity (DIO) by interfering with the function of key energy regulating neurons such as those that express the peptide proopiomelanocortin (POMC). We reported that in rats and mice, the increase in hypothalamic inflammatory gene expression occurred within 3 days of HFD feeding during a period of both excessive energy intake and hypothalamic leptin resistance. This inflammatory response in both rats and mice was accompanied by gliosis (an accumulation of enlarged microglia and reactive astrocytes) within the hypothalamus, a histopathological hallmark of acute neuronal injury. Indeed, the injury marker hsp72 was detectable in energy-regulating neurons of the mediobasal hypothalamus within 7 days of HFD exposure, and long-term consumption of the HFD eventually resulted in a 25% reduction in POMC neuron number. These results raised the possibility that the early gliosis response was initiated as a protective countermeasure to ongoing neuronal stress. To test this hypothesis, we infused the microglial inhibitory compound minocycline into the CNS of rats fed HFD or chow for 2 weeks. The animals treated with minocycline that received HFD showed an increase in food intake on days 5-8 and a 10-20% increase in body weight from day 7 onward while the drug had no effect on chow-fed rats. These data suggest that suppressing microglial activation during the early inflammatory phase of HFD feeding altered the protective gliosis response resulting in exaggerated hyperphagia and weight gain. Collectively, these results support an [ldquo]acute neuronal injury[rdquo] model of DIO, in which the defense of elevated body weight results from injury to neurons responsible for energy homeostasis, and raise the possibility that interventions to avert or reverse neuronal damage have important potential in obesity treatment and prevention.[br][br]Sources of Research Support: NIH Career Development Award DK088872 and Diabetes Endocrinology Research Center(DERC) Pilot and Feasibility Award DK017047 to JPT;NIH Grants DK068384, DK083042 and DK052989 to MWS.[br][br]Nothing to Disclose: JPT, C-XY, SG, BH, MM, HN, GJM, BEW, DEB, MT, MWS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2034 177 1144 SUN-87 PO27-01 Sunday 939 2012


938 ENDO12L_SUN-88 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Fasting-Induced Alterations in the [alpha]-MSH- and AGRP-Immunoreactive Innervation of Thyrotropin-Releasing Hormone (TRH)-Synthesizing Neurons in the Hypothalamic Paraventricular Nucleus (PVN) Andrea Kadar, Edith Sanchez, Balazs Gereben, Ronald M Lechan, Csaba Fekete Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary; Tufts Medical Center, Boston, MA; Instituto Nacional de Psiquiatria Ramon de la Fuente Muniz, Mexico City, DF, Mexico The activation of POMC neurons in the arcuate nucleus plays important role in the development of refeeding induced satiety (Singru et al., Endocrinology 2006). These anorexigenic neurons innervate neurons in the ventral parvocellular subdivision of the hypothalamic paraventricular nucleus (PVNv) that relay satiety information to the brainstem, as well as TRH neurons in the medial and periventricular parvocellular subdivisions that regulate thyroid function. Neurons of the PVNv are activated 2h after the onset of refeeding, but hypophysiotropic TRH neurons are activated only 24h after food has been reintroduced, coincident with recovery of circulating levels of thyroid hormone. To determine whether fasting-induced changes in the neuronal input to TRH neurons contribute to melanocortin resistance of these neurons during the early phase of refeeding. For this reason, we performed triple-labeling immunofluorescence on double-transgenic TRH/Cre-Z/EG mice to study whether the number of [alpha]-MSH- and AGRP-immunoreactive axon varicosities contacting hypophysiotropic TRH neurons differ under fed and fasted conditions.[br]A dense network of AGRP- and [alpha]-MSH-immunoreactive axons was found in hypothalamic paraventricular nucleus surrounding TRH neurons in both fed and fasted mice. By confocal microscopic analyses, 30.9[plusmn]0.3 AGRP-containing boutons and 19.3[plusmn]0.4 [alpha]-MSH-containing boutons were found on the surface of the perikarya and first order dendrites of the TRH neurons in the PVN of fed mice. Fasting caused a significant increase in the number of AGRP-IR varicosities on the surface of these cells (44.9[plusmn]0.3, P[lt]0.001), but decreased the number of [alpha]-MSH-IR varicosities (13.6[plusmn]0.7, P[lt]0.001). Therefore, fasting resulted in a more than two-fold increase in the ratio between AGRP and [alpha]-MSH-IR varicosities on the surface of TRH neurons (Fed vs. fasted: 1.6[plusmn]0.02 vs. 3.3 [plusmn] 0.2; P[lt]0.001).[br]Since AGRP is a potent melanocortin receptor antagonist, we hypothesize that the fasting- induced increase in the ratio between the AGRP and [alpha]-MSH-containing input to hypophysiotropic TRH neurons contributes to the delayed recovery of these neurons and the thyroid axis during the early phase of refeeding.[br][br]Singru PS et al., Endocrinology 2007;148(2):638-46.Epub 2006 Oct 26.[br][br]Sources of Research Support: OTKA K81845; EU FW7 Health-F2-2010-259772; Lend[uuml]let Award of the Hungarian Academy of Sciences; NIH DK-37021.[br][br]Nothing to Disclose: AK, ES, BG, RML, CF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1441 177 1145 SUN-88 PO27-01 Sunday 940 2012


939 ENDO12L_SUN-89 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) The Role of Ghrelin in Reward-Based Feeding Veronique St-Onge, Alfonso Abizaid Carleton University, Ottawa, Canada Combined with an increasingly sedentary lifestyle, overconsumption of preferred foods (i.e., reward-based consumption that exceeds metabolic needs) may be partly responsible for the current obesity epidemic. Recent studies suggest that ghrelin, a gut-derived orexigenic hormone, increases the incentive value of such energy-dense foods. This can lead calorically sated animals to behave like hungry ones, thereby causing weight gain. In the current study, the role of ghrelin signaling in the [ldquo]dessert phenomenon[rdquo] was assessed by comparing ghrelin receptor knock out rats (KO) to rats possessing an identical genetic background, with an intact ghrelin receptor gene (wildtype rats [ndash] WT) in a paradigm designed to model this behavior. Specifically, we measured KO and WT rats[apos] tendency to eat a palatable snack (cookie dough) when calorically sated. Ten rats of each strain were exposed to a scheduled meal regimen during which free access to food was restricted to 4 hours per day (from 10 am to 2 pm) until they consumed most of their baseline daily intake during this interval. On the last day, each rat was exposed to 30 grams of cookie dough during the last hour of the meal. While regular chow consumption was similar in both strains throughout the experiment, KO rats consumed significantly less cookie dough per grams of body weight immediately following their last meal relative to WT rats. KO rats also exhibited reduced locomotor activity prior to the meals when compared to their WT counterparts. The results obtained thus far support the idea that ghrelin is involved in both reward-based feeding and food entrainment [ndash] the natural shift in food anticipatory behavior preceding scheduled meals.[br][br]Nothing to Disclose: VS-O, AA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 273 177 1146 SUN-89 PO27-01 Sunday 941 2012


940 ENDO12L_SUN-90 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Amelioration of Binge Eating by Nucleus Accumbens Deep Brain Stimulation in Mice: A Novel Therapeutic Approach in Obesity Casey H Halpern, Jeffrey G Keating, Casey M Siegel, Tracy L Bale University of Pennsylvania, Philadelphia, PA Introduction: Altered dopaminergic transmission in the nucleus accumbens (NAc) has been implicated in binge eating disorder. Binge eating is a very common feature of obesity and contributes to the treatment-refractory nature of this condition. Here, we test the hypothesis that deep brain stimulation (DBS) of the NAc would attenuate binge eating in mice by modulating the dopaminergic system. Moreover, we hypothesized that the effect of DBS would be mediated specifically by the dopamine type 2 receptor (D2R), given its well-known implications in binge eating and obesity.[br]Methods: Mice (all 8-week males; C57Bl/6) were implanted unilaterally in the NAc (confirmed post-mortem) with a bipolar electrode (FHC). Implanted mice and non-surgical controls were provided limited daily exposure (1h) to a high fat (HF) diet (60% fat). A binge was defined as consumption of greater than or equal to 25% of daily caloric intake. Once a stable level of binge eating was measured, DBS was administered on alternate days (DBS-on; 160 Hz, 60 [mu]s, 150 [mu]A) immediately prior to and during exposure to the HF food. Consumption of this diet was examined on both DBS-on and DBS-off days. c-Fos protein immunoreactivity was then measured in the NAc and infralimbic cortex in response to DBS. A subgroup of these mice undergoing NAc DBS were pretreated with an intraperitoneal injection of a D1R-antagonist (SCH-23390; 0.075mg/kg) or a D2R-antagonist (raclopride; 3mg/kg).[br]Results: Binge eating was suppressed on DBS-on days compared to non-surgical controls (p[lt]0.05) and DBS-off days (p[lt]0.01). c-Fos-IR was increased in the NAc ipsilateral to the DBS electrode (p[lt]0.04), and there was a trend towards an increase contralaterally. No increases in c-Fos-IR were seen in the infralimbic cortex. Pretreatment with raclopride attenuated amelioration of binge eating by DBS by 51.4 percent, a difference, which differed from vehicle and SCH-23390 (p[lt]0.02). No difference was found between SCH-23390 and vehicle.[br]Conclusions: NAc DBS significantly attenuated binge eating, and was associated with increased neuronal activation. DBS amelioration of binge eating appeared to be mediated by the D2R. Taken together, these findings further implicate the dopaminergic system in binge eating, and provide evidence of opportunities for clinical translation.[br][br]Sources of Research Support: T32DA022605-05.[br][br]Nothing to Disclose: CHH, JGK, CMS, TLB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1821 177 1147 SUN-90 PO27-01 Sunday 942 2012


941 ENDO12L_SUN-91 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Mineralocorticoid Receptor Overexpressing Mice Are Resistant to High Fat Diet-Induced Obesity: Implication for the Pathophysiology of Metabolic Syndrome Emmanuelle Kuhn, Damien Le Menuet, Vixra Keo, Adeline Muscat, Geri Meduri, Say Viengchareun, Bruno Feve, Marc Lombes Faculty Medicine Paris SUd, Le Kremlin Bic[ecirc]tre, France; Univ Pierre et Marie Curie, Paris, France It has been proposed that, besides its classical renal action, the mineralocorticoid receptor (MR), a pro-adipogenic (1) and anti-thermogenic (2) factor might be involved in the pathogenesis of metabolic syndrome. To evaluate the in vivo metabolic impact of MR, we have characterized the metabolic phenotype of wild type (WT) and of transgenic (Tg) mice overexpressing human MR under the control of its proximal promoter (3) generated in our laboratory. Eight-week-old male mice were subjected to standard chow (SC) or high fat diet (HFD) for 16 weeks. Metabolic investigations (body weight (BW), mass of perirenal, gonadal and subcutaneous fat depots, glucose and insulin tolerance tests) show that Tg mice on SC have a lower weight gain (16.5 vs 19.3 g, p[lt]0.0001) than WT animals and are very sensitive to insulin with an extended hypoglycemia period, suggesting defective counterregulatory defense mechanisms against hypoglycemia. This is likely related to inappropriate responses of the hypothalamic-pituitary-adrenal axis, consistent with a stronger MR-mediated pro-active feedback, currently under investigation. The lower weight gain of Tg mice is associated with an increased small adipocytes population, as revealed by histomorphometric analysis. Tg mice under HFD are protected against diet-induced obesity, and exhibit a significant decrease of adipose depot mass (perirenal fat: 19 vs 34[permil] (mg/g BW) p=0.008; subcutaneous fat: 35 vs 80 [permil], p=0.0003; gonadal fat: 54 vs 76 [permil], p=0.045). Perirenal adipose tissues histology reveals an increase in small and large adipocytes in Tg mice, accompanied by a strong macrophage infiltration as demonstrated by specific immunodetection (11H3), compared to WT mice. We are currently evaluating the precise phenotype of infiltrating macrophages and their type of activation. Gene expression studies performed by quantitative RT-PCR, reveal a significant decrease in PPAR[gamma]2, TNF-[alpha], [beta]3-adrenergic receptor and glucocorticoid receptor mRNA levels in perirenal adipose depots of Tg mice compared to WT mice. Altered expression of these key players in adipocyte biology suggests defects in adipogenesis and lipogenic or lipolytic abnormalities. Our results demonstrate that MR exerts a pivotal metabolic role by controlling adipocyte function and energy homeostasis. Our studies should provide useful information for the pharmacological treatment of metabolic disorders in humans.[br][br](1) Caprio M et al, FASEB J, 21: 2185-2194 (2007). (2) Viengchareun S et al, Am J Physiol Endocrinol Metab, 4: E640-9 (2001). (3) Le Menuet D et al, J Biol Chem, 42 :38911-20 (2001).[br][br]Sources of Research Support: Grants form Inserm, Univ Paris Sud and ANR. EK is recipient of a CODDIM fellowship.[br][br]Nothing to Disclose: EK, DLM, VK, AM, GM, SV, BF, ML 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 480 177 1148 SUN-91 PO27-01 Sunday 943 2012


942 ENDO12L_SUN-92 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Disruption of Cue-Potentiated Feeding in Mice Lacking Ghrelin Receptors Angela Kay Walker, Jeffrey M Zigman University of Texas Southwestern Medical Center, Dallas, TX The peptide hormone ghrelin is known to have several effects on the regulation of eating behaviors. Not only is it a potent orexigenic agent that leads to increased intake of freely available food, but it also shifts food preference towards diets rich in fat, enhances operant responding for food rewards and induces conditioned place preference for food rewards. Here, we postulated that ghrelin also can affect cue-potentiated feeding, which likely plays an important role in human eating behavior. To test this hypothesis, the performance of C57Bl6/J mice as well as of mice genetically deficient in ghrelin receptors was assessed in a novel cue-potentiated feeding protocol adapted for use in mice. Using this behavioral test, it was demonstrated that C57Bl6/J mice indeed show a cue-potentiated intake of grain-based pellets, specifically with the presentation of a positive conditioned stimulus (CS+); such was enhanced by an overnight fast. However, ghrelin receptor-deficient mice lacked a specific potentiation of feeding with the presentation of the positive conditioned stimulus, as they ate the same amount with the presentation of a negative conditioned stimulus. Furthermore, it was found that the expression levels of different immediate early genes ([italic]Homer1a, C-fos,[/italic] and [italic]Arc[/italic]) within the amygdala [ndash] a brain region previously linked to cue-potentiated feeding -- paralleled the behavioral data. Activation of the amygdala was observed with the positive cue but not the negative cue in wild-type mice, while activation was not specific to either cue in the ghrelin-receptor deficient mice. As such, we have validated a new mouse behavioral model with which to study the mechanisms controlling cue-potentiated feeding. Using this model, we have demonstrated a key role for ghrelin receptor expression in establishing a cue-association in this important food behavioral response.[br][br]Nothing to Disclose: AKW, JMZ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 187 177 1149 SUN-92 PO27-01 Sunday 944 2012


943 ENDO12L_SUN-93 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) HDC-tdTomato Reporter Mouse Model: A New Tool for Visualizing ECL Cells Won-Mee Park, Angela K Walker, Ichiro Sakata, Sherri Osborne-Lawrence, Chelsea Migura, Jeffrey M Zigman The University of Texas Southwestern Medical Center, Dallas, TX; The University of Texas Southwestern Medical Center, Dallas, TX; Saitama University, Saitama, Japan Enterochromaffin-like (ECL) cells, a subset of endocrine cells in the mucosal lining of the stomach, are the major source of gastric histamine. While ECL cells have long been known, there remains much to be learned about their function and their physiological regulation. Here, we report the validation of a new transgenic mouse line in which Cre recombinase expression has been targeted to cells expressing histidine decarboxylase (HDC), which catalyzes the rate-limiting step in the synthesis of histamine. This was achieved by crossing the HDC-Cre mouse with Rosa26-tdTomato reporter mice. As expected, the tdTomato signal was co-localized with HDC-immunoreactivity within the gastric mucosa. In particular, nearly 75% of cells with HDC-immunoreactivity contained tdTomato fluorescence, whereas nearly 100% of cells with tdTomato fluorescence contained HDC-immunoreactivity. HDC expression within tdTomato-positive cells was further confirmed by quantitative PCR analysis of the mRNA content of highly purified populations of tdTomato-positive cells generated by fluorescent activated cell sorting (FACS). We suggest that these transgenic mice will enable us to better investigate and visualize ECL cells for further studies in order to acquire deeper insight into the role they play in gastric physiology.[br][br]Sources of Research Support: This work was funded from the NIH (K08DK068069-01A2 to JMZ) and an International Research Alliance with The Novo Nordisk Foundation Center for Basic Metabolic Research (to JMZ).[br][br]Nothing to Disclose: W-MP, AKW, IS, SO-L, CM, JMZ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1589 177 1150 SUN-93 PO27-01 Sunday 945 2012


944 ENDO12L_SUN-94 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Genetic Disruption of Brain Serotonin Content in Mice Produces Leanness with Hyperphagia and Elevated Physical Activity but Minimal Skeletal Changes Robert Brommage, Deon D Smith, Jeff Liu, Wangsheng Yu, Qi M Yang, David R Powell Lexicon Pharmaceuticals, Inc, The Woodlands, TX; Lexicon Pharmaceuticals, Inc, The Woodlands, TX; Lexicon Pharmaceuticals, Inc, The Woodlands, TX Tryptophan hydroxylase (TPH) is the rate-limiting enzyme for serotonin synthesis. Distinct enzymes are involved in peripheral (TPH1) and brain (TPH2) serotonin production.[br]Yadav et al. (1) reported that 4-12 week-old Tph2 knockout (KO) mice are lean and hypophagic, with elevated physical activity and energy expenditure but reduced trabecular bone mass. We examined metabolic and skeletal parameters in Tph2 KO mice, generated by disruption of exons 1 and 2 (2). KO mice have brain serotonin levels below 15% of normal but show only minor changes in standard behavioral tests. Lean body mass and femur length are normal.[br]Multiple cohorts of Tph2 KO mice fed standard or high fat diets were lean between 10 and 42 weeks of age. In one cohort this lean phenotype was lost after 42 weeks of age, and two additional cohorts were not lean when studied at ages [gt] 42 weeks. Tph2 KO mice had increased food consumption at 50 weeks of age (19%, P = 0.001 for cohort 1 and 36%, P = 0.003 for cohort 2). Physical activity (56 and 70 weeks) was elevated during both light and dark phases in both cohorts. Consistent with hyperphagia and high activity, Tph2 KO mice had increased oxygen consumption (daytime [Delta] = 14%, P = 0.06 and nighttime [Delta] = 15%, P = 0.008).[br]Female Tph2 KO mice (39 to 51 weeks of age) responded normally to the anorexic effect of a single 25 mg/kg dose of fenfluramine, believed to activate serotonin 5HT2c receptor pathways. Selection of dietary fat (white chocolate versus purified low-fat diet) was not influenced by Tph2 disruption.[br]Male, but not female, Tph2 KO mice have slightly elevated (19% at 21 weeks, P [lt] 0.001 and 8% at 33 weeks, P = NS) spine trabecular bone mass. Female, but not male, Tph2 KO mice have slightly reduced (9% at 69 weeks, P [lt] 0.05 and 13% at 83 weeks, P [lt] 0.001) midshaft femur cortical bone thickness.[br]Studying older mice, we do not confirm findings of low trabecular bone mass reported previously (1). Young Tph2 KO mice are lean, but this phenotype is lost with age. These older mice have increased physical activity and energy expenditure as previously reported, but are hyperphagic rather than hypophagic (perhaps explaining loss of the lean phenotype). These findings suggest Tph2 KO mice have 1) a minimal bone mass phenotype as adults; and 2) a more complex metabolic phenotype than was reported initially, consistent with dysregulation of a neurotransmitter having roles in multiple CNS pathways.[br][br](1) Yadav VK et al., Cell 2009;138:976. (2) Savelieva KV et al., PLoS ONE 2008; 3:e3301.[br][br]Disclosures: RB: Employee, Lexicon Pharmaceuticals, Inc. DDS: Employee, Lexicon Pharmaceuticals, Inc. JL: Employee, Lexicon Pharmaceuticals, Inc. WY: Employer, Lexicon Pharmaceuticals, Inc. QMY: Employee, Lexicon Pharmaceuticals, Inc. DRP: Employee, Lexicon Pharmaceuticals, Inc. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1721 177 1151 SUN-94 PO27-01 Sunday 946 2012


945 ENDO12L_SUN-95 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Mapping Functional Leptin Receptors and Leptin-Activated Transcriptional Pathways in the Hypothalamus and Pituitary of the Frog [italic]Xenopus laevis[/italic] Melissa Cui, Caroline Hu, Adam Dziuba, Robert John Denver University of Michigan, Ann Arbor, MI Leptin inhibits food intake in nonmammalian species, but it is not known if the intracellular signaling pathways engaged by the leptin receptor (LepR) and the neural pathways involved in feeding controls are evolutionarily conserved. We investigated these questions using juvenile [italic]Xenopus laevis[/italic] frogs. Intracerebroventricular (i.c.v.) injection of recombinant [italic]Xenopus[/italic] leptin (rxLeptin; 20 ng/g BW) inhibited feeding in frogs. In the mouse, leptin actions on food intake depend on the LepR activating signaling by the transcription factor Signal Transducer and Activator of Transcription 3 (STAT3). Using reporter assays in Cos-7 cells we found that rxLeptin caused a dose-dependent activation of frog LepR signaling via STAT3. Mutation of the evolutionarily conserved tyrosine at position 1127 to alanine abrogated STAT3 signaling. We conducted immunohistochemistry on frog brain for phosphorylated STAT3 to determine if leptin activates STAT3 signaling [italic]in vivo[/italic]. pSTAT3 immunoreactivity (ir) was strongly increased in frog brain and pituitary 1 hr after i.c.v. injection of rxLeptin. We used pSTAT3-ir as a proxy for functional LepR to map LepR expressing cells in the frog brain and pituitary. We found robust pSTAT3-ir in the parvocellular and magnocellular divisions of the anterior preoptic area (POA), and the ventral hypothalamus/lateral hypothalamus (VH/LH) that began in the most rostral limit and extended caudally, with highest pSTAT3-ir in the most caudal extent of the VH/LH. We observed strong pSTAT3-ir throughout the anterior pituitary gland. We also found that activation of LepR signaling upregulated STAT3 target genes in frog brain [italic]in vivo. [/italic]Injection of rxLeptin i.c.v. increased Suppressor of Cytokine Signaling 3 and proopiomelanocortin mRNAs in the frog POA/hypothalamus at 2 hr as measured by RTqPCR. Microarray analysis on hypothalamus/pituitary 2 hr after leptin injection identified 141 regulated genes, 60 upregulated and 81 downregulated. Our findings show evolutionary conservation of mechanisms of LepR signaling, and the neural substrates that mediate leptin actions on food intake.[br][br]Sources of Research Support: NSF grant IOS 0641587 to RJD.[br][br]Nothing to Disclose: MC, CH, AD, RJD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2204 177 1152 SUN-95 PO27-01 Sunday 947 2012


946 ENDO12L_SUN-96 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Nesfatin-1 Attenuates the Orexigenic Effects of Ghrelin and Suppresses Endogenous Ghrelin in Fischer 344 Rats Yona R Vandersluis, Haneesha Mohan, Suraj Unniappan York University, Toronto, Canada Nesfatin-1 is a metabolic hormone with insulinotropic and anorectic effects in rats and mice. Several studies have indicated that nesfatin-1 colocalizes ghrelin, a potent stimulant of food intake. We hypothesized that nesfatin-1 would mediate satiety by inhibiting the ghrelin system, namely, ghrelin, ghrelin-o-acyltransferase (GOAT), and the ghrelin receptor. The aim of this project was to identify the ghrelin-nesfatin-1 relationship in male Fischer 344 rats. In this study, we employed a number of techniques including fluorescence immunohistochemistry, quantitative real-time PCR, sub-cutaneous administration of peptides using Alzet osmotic mini-pumps, oral gavage of a mixed meal, and measurement of metabolic parameters using the Comprehensive Laboratory Animal Monitoring System (CLAMS). We report here for the first time that nesfatin-1-like immunoreactivity (ir) colocalizes both ghrelin receptor and GOAT in the stomach cells. In order to determine whether synthetic rat nesfatin-1 affects endogenous ghrelin, we administered 100 micrograms/kg body weight synthetic rat nesfatin-1 to rats, fasted them for 16 hours, provided 2 mL of vanilla flavoured Ensure Plus as oral gavage, and measured ghrelin and GOAT mRNA expression in the stomach. Nesfatin-1 infusion caused a significant reduction in preproghrelin mRNA expression in the stomach at 210 minutes post-gavage. GOAT mRNA expression in the stomach was also significantly reduced in nesfatin-1 treated rats, but this effect was found at an earlier time point (120 minutes post-gavage). We then studied whether nesfatin-1 influences the orexigenic effects of exogenous ghrelin. Sub-cutaneous infusion of synthetic rat ghrelin (400 micrograms/kg body weight) caused [sim]40% increase in both the dark phase cumulative food intake and duration of feeding bouts (measured using the CLAMS). Nesfatin-1 (100 micrograms/kg body weight) infusion inhibited basal food intake during the dark phase. Co-infusion of both nesfatin-1 with ghrelin at the doses mentioned above caused no changes in feeding, indicating a loss of feeding regulatory activities of both peptides. Together, our results provide the first line of data to implicate ghrelin as a potential target that mediates the metabolic effects of nesfatin-1. Further studies are required to address whether ghrelin is critical in eliciting the metabolic effects of nesfatin-1 in rats.[br][br]Sources of Research Support: This work is supported by the Canadian Institutes of Health Research (CIHR) operating grant, and new investigator salary award to SU.[br][br]Nothing to Disclose: YRV, HM, SU 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1659 177 1153 SUN-96 PO27-01 Sunday 948 2012


947 ENDO12L_SUN-97 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Ghrelin Receptor Regulates Meal Patterns during Aging Ligen Lin, Xiaojun Ma, Yuxiang Sun Baylor College of Medicine, Houston, TX Ghrelin is the only circulating orexigenic hormone. It initiates meals, stimulates food intake, and increases adiposity through its receptor, Growth Hormone Secretagogue Receptor (GHS-R). Our previous studies showed that despite the fact that the Ghsr-null mice have the same total daily food intake and physical activity as wild-type (WT) mice, older Ghsr-null mice have reduced body weight and adiposity, and increased thermogenesis (1). Moreover, the null mice exhibited many youthful characteristics, such as higher resting metabolic rate, improved lipid profiles and enhanced insulin sensitivity.[br]Meal patterns have been suggested to affect fasting lipids, postprandial insulin profiles and thermogenesis, but feeding behavior during aging is largely unexplored. In the present study, we monitored food intake and meal patterns of young (4-5 months), middle-aged (10-12 months) and old (18-24 months) WT and Ghsr-null mice, using an indirect calorimetry system. We made the following interesting observations: 1) Comparing different age groups of WT mice, middle-aged and old mice consumed smaller meals when compared with young animals, but daily meal frequency did not change. 2). Comparing Ghsr-null mice with WT mice, although Ghsr-null mice have same total daily food intake as WT mice, the null mice ate larger meals, at a lower meal frequency and with longer meal duration (ate fewer and bigger meals, but did not actually eat more). Hypothalamic neuropeptides are known to regulate food intake and meal size. During aging, orexigenic neuropeptide Y (NPY) and Agouti-related peptide (AgRP) expression was decreased in hypothalamus, but anorexic pro-opiomelanocortin (POMC) expression remained unchanged. Intriguingly, GHS-R ablation increased NPY/AgRP expression during aging, but decreased POMC expression. The hypothalamic gene expression profile exhibited by the Ghsr-null mice is similar to that of calorie-restricted WT animals. These results collectively suggest that age-associated anorexia is primarily associated with reduced meal size. GHS-R regulates meal patterns: GHS-R ablation results in larger average meal size, lower meal frequency, and longer meal duration; this may have beneficial effects on energy metabolism during aging. GHS-R antagonists may represent a new means of combating obesity by regulating meal patterns.[br][br]1. Aging Cell, 10: 996[ndash]1010.[br][br]Sources of Research Support: We gratefully acknowledge the support of NIH/NIA grant 1R03AG029641-01 (YS), USDA/CRIS grant ARS 6250-51000-055 (YS), and the NIH-Diabetes and Endocrinology Research Center (P30DK079638) at Baylor College of Medicine.[br][br]Nothing to Disclose: LL, XM, YS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2254 177 1154 SUN-97 PO27-01 Sunday 949 2012


948 ENDO12L_SUN-98 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Nesfatin-1 Neurons in the Hypothalamic Paraventricular Nucleus Are Regulated by Meal-Evoked Factors Darambazar Gantulga, Yuko Maejima, Masanori Nakata, Toshihiko Yada School of Medicine, Jichi Medical University, Shimotsuke, Japan; National Institute for Physiological Sciences, Okazaki, Japan Nesfatin-1, a recently discovered anorectic peptide processed from NUCB2, is expressed in the neurons in specific brain areas including the paraventricular nucleus of hypothalamus (PVN), a pivotal center of feeding regulation. Although nesfatin-1[apos]s downstream anorectic neural pathway has been well investigated, the regulation of nesfatin-1 expressing neurons in the PVN has little been studied. Since starvation decreases and refeeding stimulates nesfatin-1 expression specifically in the PVN, we examined direct effects of meal-evoked metabolic factors, glucose, insulin and cholecystokinin (CCK), on PVN nesfatin-1 neurons.[br]We prepared brain slice and dissected PVN from C57BL/6 mice, isolated single neurons by enzyme treatment combined with trituration, and measured cytosolic calcium concentration ([Ca[sup]2+[/sup]][sub]i[/sub]) by ratiometric fura-2 fluorescence imaging. The effects of glucose, insulin and CCK on [Ca[sup]2+[/sup]][sub]i[/sub] were measured in isolated single PVN neurons, followed by immunocytochemical staining of these neurons for nesfatin-1.[br]Glucose (10 mM), insulin (10[sup]-13[/sup]M) and CCK (10[sup]-13[/sup]M) induced increases in [Ca[sup]2+[/sup]][sub]i [/sub]in 55 of 311 (16.6%), 32 of 249 (12.9%) and 8 of 39 (20.5%) PVN neurons tested, respectively. Moreover, the post [Ca[sup]2+[/sup]][sub]i[/sub] measurement immunocytochemistry identified 58% of glucose-responsive neurons, 63% of insulin-responsive neurons, and 50% of CCK-responsive neurons as nesfatin-1 neurons, indicating that nesfatin-1 neuron is the major target for glucose, insulin and CCK in the PVN.[br]Our data demonstrate that glucose, insulin and CCK directly interact with and increase [Ca[sup]2+[/sup]][sub]i[/sub] in nesfatin-1 neurons in the PVN, and that the nesfatin-1 neuron serves as the major target for all of them in the PVN. Postprandial activation of the PVN nesfatin-1 neurons could be mediated by these meal-evoked nutritional and endocrine factors, which potentially contributes to generation of satiety.[br][br]Nothing to Disclose: DG, YM, MN, TY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 197 177 1155 SUN-98 PO27-01 Sunday 950 2012


949 ENDO12L_SUN-99 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Omentin-1 Stimulates Food Intake in Rats Luigi Brunetti, Lucia Recinella, Annalisa Chiavaroli, Sheila Leone, Claudio Ferrante, Fabio Manippa, Michele Vacca G d[apos]Annunzio University, Chieti, Italy [bold]Context: [/bold]Omentin is a visceral fat depot-specific secretory protein [1], and omentin-1, its main circulating isoform, is inversely correlated with obesity and insulin resistance [2].[br][bold]Objectives: [/bold]We aimed to investigate the effects of omentin-1 on feeding behavior and the hypothalamic gene expression of the orexigenic peptides agouti-related peptide (AgRP), neuropeptide Y (NPY), orexin-A,and the anorexigenic cocaine- and amphetamine-regulated transcript (CART) peptide, corticotropin releasing hormone (CRH), proopiomelanocortin (POMC), in male Wistar rats fed a standard laboratory diet. We also evaluated the effects of omentin-1 on dopamine (DA), norepinephrine (NE) and serotonin (5-hydroxytryptamine, 5-HT) release from rat hypothalamic neuronal endings (synaptosomes) [italic]in vitro[/italic].[br][bold]Design:[/bold] 10 animals (5 for each group of treatment) were injected intraperitoneally, at 9.00 am, daily for 14 days with either omentin-1 (1.5 [mu]g/rat) or saline. 24 h after each injection, daily food intake and body weight were recorded. Total RNA was extracted from hypothalami and reverse transcribed to evaluate peptide gene expression by real-time reverse transcription polymerase chain reaction. Synaptosomes were obtained as previously described [3]. They were loaded with either [[sup]3[/sup]H]DA, [[sup]3[/sup]H]NE or [[sup]3[/sup]H]5-HT and perfused with graded concentrations (1-100 ng/ml) of omentin-1, both basally and during K[sup]+[/sup](15 mM)-induced depolarization. Food intake, body weight and gene expression data were analyzed by unpaired t test. In the synaptosome perfusion experiments, results were analyzed by one-way analysis of variance (ANOVA), followed by Newman-Keul[apos]s multiple comparison test.[br][bold]Results: [/bold]Compared to vehicle, omentin-1 significantly increased food intake since day 10 of treatment (days 10-13, [italic]P[/italic][lt]0.05; day 14, [italic]P[/italic][lt]0.01) and body weight on day 14 ([italic]P[/italic][lt]0.05). Omentin-1 treatment led to a significant reduction in both CART ([italic]P[/italic][lt]0.05) and CRH ([italic]P[/italic][lt]0.05) gene expression, while POMC, AgRP, NPY and orexin-A gene expression were not modified with respect to vehicle-treated rats. In hypothalamic synaptosomes, omentin-1 (10-100 ng/ml) stimulated basal NE release (ANOVA, [italic]P[/italic][lt]0.0001; post-hoc, [italic]P[/italic][lt]0.001 vs. vehicle), in a dose-dependent manner, leaving unaffected either basal or depolarization-induced DA and 5-HT release.[br][bold]Conclusions: [/bold]The orexigenic effects of omentin-1 could be partially related to decreased CART and CRH gene expression and increased NE release in the hypothalamus.[br][br][1] Yang RZ et al., Am J Physiol Endocrinol Metab 2006; 290:E1253. [2] de Souza Batista CM et al., Diabetes 2007; 56:1655. [3] Brunetti L et al., Eur J Pharmacol 1999; 372:237.[br][br]Sources of Research Support: Italian Ministry of Education grants.[br][br]Nothing to Disclose: LB, LR, AC, SL, CF, FM, MV 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 337 177 1156 SUN-99 PO27-01 Sunday 951 2012


950 ENDO12L_SUN-100 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Effects of Growth Hormone-Releasing Hormone (GHRH) Gene Ablation on Hypothalamic Peptide Gene Expression in Mice Luigi Brunetti, Rugia Shohreh, Chiara Di Nisio, Giustino Orlando, Roberto Salvatori, Michele Vacca G d[apos]Annunzio University, Chieti, Italy; Johns Hopkins University School of Medicine, Baltimore, MD [bold]Context:[/bold] Growth hormone deficiency (GHD) induces growth failure and alterations in body composition in both humans and mice [1]. GHD is associated with decreased lean body mass, increased fat deposition and central obesity in late adulthood [2]. Mice with GH-releasing hormone (GHRH) gene ablation [GHRH knockout (GHRHKO)] show GHD and pituitary hypoplasia [3]. Adult GHRHKO animals appear to be obese compared to mice homozygous for the normal allele (+/+) [2].[br][bold]Objectives:[/bold] Our study aimed to investigate the gene expression of hypothalamic peptides which play a key role in feeding behavior in adult mice homozygous for GHRHKO allele ([minus]/[minus]). Adult mice heterozygous for GHRHKO allele (+/[minus]) were used as control because no significant difference in body weight was observed between +/+ and +/[minus] animals from the 2[sup]nd[/sup] to the 12[sup]th[/sup] week of life [3].[br][bold]Design:[/bold] 6 +/[minus] and 8 [minus]/[minus] mice were sacrificed at 12 weeks of age. Total RNA was extracted from hypothalami and reverse transcribed to evaluate the gene expression of the orexigenic peptides agouti-related peptide (AgRP), neuropeptide Y (NPY) and orexin-A, as well as the anorexigenic cocaine- and amphetamine-regulated transcript (CART), corticotrophin releasing hormone (CRH) and proopiomelanocortin (POMC) by real-time reverse transcription polymerase chain reaction (real-time RT PCR). Gene expression data, deriving from relative quantification, were analyzed by one-sample t test. 1 (calibrator sample, control) was obviously considered the theoretical mean for the comparison. The level of statistical significance was set as [italic]P[/italic][lt]0.05.[br][bold]Results:[/bold] Adult [minus]/[minus] mice show a significant increase in AgRP mRNA levels ([italic]P[/italic]=0.0213) and a significant reduction in both CART ([italic]P[/italic]=0.0164) and CRH ([italic]P[/italic][lt]0.0001) gene expression compared to adult +/[minus] mice. On the other hand, we did not observe any significant difference in NPY, orexin-A and POMC mRNA levels between adult +/[minus] and [minus]/[minus] mice.[br][bold]Conclusions:[/bold] We can therefore hypothesize that the obese phenotype of GHRHKO mice is driven by increased AgRP gene expression and inhibited CART and CRH mRNA levels in the hypothalamus.[br][br][1] Alba M, Fintini D, Salvatori R (2005). Effects of recombinant mouse growth hormone treatment on growth and body composition in GHRH knockout mice. Growth Horm IGF Res 15:275-282. [2] Alba M, Schally AV, Salvatori R (2005). Partial reversibility of growth hormone (GH) deficiency in the GH-releasing hormone (GHRH) knockout mouse by postnatal treatment with a GHRH analog. Endocrinology 146:1506-1513. [3] Alba M, Salvatori R (2004). A mouse with targeted ablation of the growth hormone-releasing hormone gene: a new model of isolated growth hormone deficiency. Endocrinology 145:4134-4143.[br][br]Sources of Research Support: Italian Ministry of Education grants.[br][br]Nothing to Disclose: LB, RS, CDN, GO, RS, MV 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 339 177 1157 SUN-100 PO27-01 Sunday 952 2012


951 ENDO12L_SUN-101 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Studies of Hypothalamic Murine Melanocortin 3 Receptor ([italic]MC3R)[/italic] Expression Reveal Two Major Transcriptional Start Sites (TSSs) and Splicing in the 3[prime] Untranslated Region (UTR) Arunabha Basu, Xin Wen, Sender Lkhagvadorj, Jack A Yanovski National Institute of Child Health and Human Development, Bethesda, MD [bold]Background:[/bold] The melanocortin 3 receptor ([italic]MC3R[/italic]), a single-exon gene expressed in hypothalamic and peripheral tissues, is involved in regulation of energy homeostasis. However, [italic]MC3R[/italic] transcript structure is not well understood. We therefore studied initiation and termination sites for hypothalamic murine [italic]MC3R[/italic] transcripts.[br][bold]Methods:[/bold] RNA Ligase Mediated Rapid Amplification of cDNA Ends (RLM-RACE) was conducted on the 5[apos] end of DNase-treated total murine hypothalamic RNA. A separate RACE was done for the 3[apos] end of the total RNA. The products of each RACE were amplified through nested PCR using primers for a ligated adapter and gene-specific primers for 5[apos]RACE and a polyT-linked primer plus gene-specific primers for 3[apos]RACE. PCR products were cloned into plasmids and sequenced.[br][bold]Results:[/bold] From 92 PCR products sequenced from four independent 5[apos] RACE experiments, we found 24 clones had a TSS [sim]368 bases upstream of the start of gene translation, at the predicted TSS according to GenBank. Another 46 sequences had a TSS [sim]439 bases upstream of the ATG, where another potential TSS, with an initiator sequence and an associated CpG Island, was identified by TFBIND and the CpG Island Searcher. Confirmatory data for both TSSs were found among sequences obtained from RLM-RACE of non-DNAse treated mRNA. From 42 sequences obtained from two independent 3[apos] RACE experiments, 29 clones terminated [sim]1,200 bases after the translational stop codon. A 787 base splice between a consensus splice donor and a splice acceptor site was observed in 27 of these 29 sequences. Another 13 sequences terminated between 500 and 700 bases after the stop codon.[br][bold]Conclusions:[/bold] Mouse hypothalamus expresses multiple [italic]MC3R[/italic] transcripts. Two major TSSs and multiple sites of transcriptional termination were identified. These data suggest the possibility that tissue specificity for, and regulation of, [italic]MC3R[/italic] expression might be achieved through selection of transcripts. Mechanisms involved in the selection of these multiple TSSs and termination sites need to be determined. Ongoing studies will examine variation in expressed [italic]MC3R[/italic] transcripts in other mouse tissues as well as determine the structure of human [italic]MC3R[/italic] transcripts in hypothalamic and peripheral tissues.[br][br]Sources of Research Support: This study was supported by the Intramural Research Program of the National Institute of Child Health and Human Development and National Institute of Nursing Research, NIH.[br][br]Nothing to Disclose: AB, XW, SL, JAY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1938 177 1158 SUN-101 PO27-01 Sunday 953 2012


952 ENDO12L_SUN-102 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Leptin Regulation of cAMP Levels in the Arcuate Nucleus and Protein Kinase B in the Hypothalamus Is Impaired during the Development of Diet-Induced Obesity Abhiram Sahu, Maitrayee Sahu University of Pittsburgh, Pittsburgh, PA Phosphatidyinositol 3-Kinase (PI3K)-phosphodiesterase-3B (PDE3B)-cAMP pathway plays an important role in transducing leptin action in the hypothalamus (1). Obesity is usually associated with hyperleptinimia and leptin resistance. Rodent models of diet-induced obesity (DIO), in which animals become obese and hyperleptinimic with a high fat diet (HFD), appear to provide compelling models for human obesity. Although central leptin resistance is thought to be involved in the development of DIO, the mechanism behind this phenomenon is not fully understood. Since hypothalamic PI3K-PDE3B pathway of leptin signaling is impaired in DIO mice (2,3), we tested if leptin regulation of hypothalamic cAMP pathway was impaired during the development of DIO. We also tested if leptin stimulated phosphorylation of protein-kinase B (p-AKT) in the hypothalamus and whether this regulation was altered in DIO mice. To this end, 4-wk-old male FVB/N mice were fed with either a low-fat diet (LFD, 6% kcal as fat] or a HFD (58% kcal as fat) for a period of 4 wk. At the end of dieting, the animals were fasted overnight, followed by leptin (3.5 mg/kg, ip) or saline injection. Thirty minutes later, animals were killed. Brains were dissected out, frozen on dry ice, sectioned on a cryostat and hypothalamic nuclei (arcuate, VMN, DMN, LH, PVN) were microdissected for cAMP determination by ELISA kit. In some animals, the medial basal hypothalami (MBH) were dissected out to measure p-AKT levels by Western blotting. The results showed increased body wt gain in HFD-fed group as compared to LFD-fed group in association with increased fat pad weights and food intake, suggesting the development of DIO in HFD-fed mice. Results on cAMP levels in the microdissected hypothalamic nuclei showed that leptin selectively decreased cAMP levels in the ARC of LFD-fed mice. However, leptin had no effect on cAMP levels in the ARC or any other nuclei in HFD-fed mice. In addition, whereas leptin significantly increased p-AKT levels in the MBH of LFD-fed mice, it failed to do so in the MBH of HFD-fed mice. In contrast, leptin increased hypothalamic p-STAT3 levels in both LFD- and HFD-fed mice. These results suggest impairment in leptin regulation of cAMP levels in the ARC and AKT-pathway in the hypothalamus during the development of DIO. In conclusion, a defect in leptin regulation of cAMP pathway and AKT signaling in the hypothalamus may be one of the mechanisms of central leptin resistance and DIO.[br][br](1) Zhao et al., Nature Neuroscienc 2002; 5:727. (2) Metlakunta et al., Endocrinology 2008; 149:1121. (3) Sahu and Sahu, Abst. 93rd Ann. Meet. Endo. Soc.2011;# P2-287.[br][br]Sources of Research Support: NIH Grant DK78068 awarded to AS.[br][br]Nothing to Disclose: AS, MS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 641 177 1159 SUN-102 PO27-01 Sunday 954 2012


953 ENDO12L_SUN-103 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Glucose-Sensing Mechanisms Involved in the Downregulation of AgRP Gene Expression Using Immortalized Hypothalamic Glucose-Inhibitory Cell Models Janet J Jang, Jennifer A Chalmers, Shelinna Xu, Mary-Ellen Harper, Denise D Belsham University of Toronto, Toronto, Canada; Toronto General Research Institute, University of Health Network, Toronto, Canada; Faculty of Medicine, University of Ottawa, Ottawa, Canada Glucose-sensing (GS) neurons of the hypothalamus are known to play a key role in the regulation of energy homeostasis. Classified as either glucose excited (GE) or glucose inhibited (GI) neurons, they largely coincide with neurons that express anorexigenic (POMC) or orexigenic (NPY/AgRP) neuropeptides, respectively. While the mechanisms by which GE neurons respond to increased glucose via upregulation of anorexigens are more established, the intracellular signalling mechanisms by which GI neurons decrease expression and secretion of orexigenic neuropeptides are not well understood. We have generated an array of immortalized hypothalamic cell lines that can be used for these studies, the clonal embryonic mHypoE29/1 and the adult mHypoA-NPY/GFP, both expressing NPY/AgRP. Using semi-quantitative RT-PCR, we demonstrated that both cell lines show strong expression of glucose transporters (GLUT1,3,4,5), phosphofructokinases (PFK A,B,C), and hexokinases (HK1,2) required for the uptake and metabolism of glucose. These cell models lack expression of components involved in GE glucose-sensing, such as the ATP-sensitive potassium channels, and glucokinase. To assess the metabolic responses of these neurons to changes in glucose, we used the Seahorse Extracellular Flux (XF) Analyzer (Seahorse Biosciences). Measurements of oxygen consumption rate (OCR) revealed a significant decrease in respiratory capacity when exposed to increasing concentrations of glucose, mHypoE29/1 (p[lt]0.01) and mHypoA-NPY/GFP (p[lt]0.05), suggesting a quiescence in metabolic activity expected of GI neurons. Using real-time RT-PCR, we demonstrated that following overnight glucose restriction, treatment with 0.5, 2.5, 5.0 mM extracellular glucose results in a 30-40% downregulation of AgRP mRNA within 1 h (p[lt]0.05). Only the highest glucose treatment of 5.0 mM was able to further maintain the repression of AgRP mRNA for up to 2 h. We are currently characterizing calcium responsiveness, changes in reactive oxygen species levels (ROS), and investigating the role of the AMP-activated protein kinase pathway in modulating the changes in AgRP mRNA expression and secretion. Our evidence suggests that these neuronal cell models possess the characteristics of GI glucose sensing neurons of the hypothalamus, and may therefore provide insight into the cellular mechanisms underlying glucose sensing in orexigenic neurons.[br][br]Sources of Research Support: This research is supported by CIHR, CFI, CRC, and BBDC.[br][br]Nothing to Disclose: JJJ, JAC, SX, M-EH, DDB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 838 177 1160 SUN-103 PO27-01 Sunday 955 2012


954 ENDO12L_SUN-104 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Acutely Decreased Thermoregulatory Energy Expenditure or Decreased Activity Energy Expenditure Both Acutely Reduce Food Intake in Mice Karl J Kaiyala, Gregory J Morton, Joshua P Thaler, Thomas H Meek, Tracy Tylee, Kayoko Ogimoto, Brent E Wisse University of Washington, Seattle, WA; University of Washington, Seattle, WA Despite the suggestion that reduced energy expenditure (EE) is a key contributor to the obesity pandemic, few studies have tested whether reduced EE is associated with a compensatory reduction in food intake (FI). The homeostatic mechanisms that control food intake (FI) and energy expenditure (EE) remain controversial and are thought to act over days to weeks. We evaluated FI in mice using two models of acutely decreased EE: 1) increasing ambient temperature to thermoneutrality in mice acclimated to standard laboratory temperature or 2) exercise cessation in mice accustomed to wheel running.[br]Increasing ambient temperature (from 21[ordm]C to 28[ordm]C) rapidly decreased EE, demonstrating that thermoregulatory EE contributes to both light cycle (40[plusmn]1%) and dark cycle EE (15[plusmn]3%) at normal ambient temperature (21[ordm]C). Reducing thermoregulatory EE acutely decreased FI primarily during the light cycle (65[plusmn]7%), thus conflicting with the delayed compensation model, but did not alter spontaneous activity. Acute exercise cessation decreased EE only during the dark cycle (14[plusmn]2% at 21[ordm]C; 21[plusmn]4% at 28[ordm]C), while FI was reduced during the dark cycle (0.9[plusmn]0.1g) in mice housed at 28[ordm]C, but during the light cycle (0.3[plusmn]0.1g) in mice housed at 21[ordm]C. Cumulatively, there was a strong correlation between the change in daily EE and the change in daily FI (R2=0.51, p[lt]0.001).[br]We conclude that acutely decreased EE decreases FI suggesting that energy intake is regulated by metabolic signals that respond rapidly and accurately to reduced EE.[br][br]Nothing to Disclose: KJK, GJM, JPT, THM, TT, KO, BEW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 360 177 1161 SUN-104 PO27-01 Sunday 956 2012


955 ENDO12L_SUN-105 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Differential Expression of mRNA, microRNA and Protein during Experimentally Induced Changes in Hypothalamic Energy Homeostasis Hao Jiang, Thero Modise, Robert Bowen, Richard Helm, Roderick V Jensen, Deborah J Good Virginia Tech, Blacksburg, VA; Virginia Tech, Blacksburg, VA; Virginia Tech, Blacksburg, VA; Virginia Tech, Blacksburg, VA An imbalance between energy intake and energy expenditure in humans leads to obesity, a major health problem in the US, with nearly 60% of the US adult population either obese or overweight. As the hypothalamus is a primary site of energy balance control, an understanding the fundamental responses of this brain tissue in energy homeostasis is of clinical, pharmaceutical, and societal importance. To further clarify hypothalamic energy homeostasis control mechanisms, protein, mRNA, and microRNA levels from [italic]ad lib[/italic] fed and food deprived mice were assessed by mass spectrometry-based proteomics and transcriptional profiling. A multi-dimensional chromatographic separation and tandem mass spectrometric analysis identified nearly 200 hypothalamic proteins, with several of these proteins found to be differentially expressed between [italic]ad lib[/italic] fed and food deprived mice. The mRNA and microRNA studies revealed that more than 1000 genes were expressed at different levels between the two groups, and nearly 30 microRNAs appeared to be differentially expressed. Several mRNAs and microRNAs were also selected for quantitative PCR analysis to confirm their regulation by Taqman assay. mRNAs associated with cell fate have consistent differences between the two energy availability states. While no selected microRNAs showed significant changes between two groups to date, and the correlation between changes at the transcriptional and protein levels was low, identified gene targets can be used to characterize novel pathways in energy homeostasis. Overall, our data suggest that differential expression of proteins, mRNAs, and microRNAs in the hypothalamus are related to the balance between energy intake and energy expenditure and that further understanding of the root mechanisms involved in the hypothalamic response to energy imbalance will require assessment at both the transcript and protein levels.[br][br]Nothing to Disclose: HJ, TM, RB, RH, RVJ, DJG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 770 177 1162 SUN-105 PO27-01 Sunday 957 2012


956 ENDO12L_SUN-106 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Proglucagon Gene Expression Is Regulated by Insulin, Leptin and cAMP in Mouse Embryonic and Adult Hypothalamic Neuronal Cells Prasad S Dalvi, Frederick D Erbiceanu, David M Irwin, Denise D Belsham University of Toronto, Toronto, Canada; University of Toronto, Toronto, Canada; University of Toronto, Toronto, Canada; University of Toronto, Toronto, Canada; University of Toronto, Toronto, Canada Proglucagon-derived peptides (PGDPs) play important roles in energy homeostasis and appetite regulation. The proglucagon gene is expressed in islet [alpha]-cells and intestinal L-cells, as well as a limited number of hypothalamic neurons. The two key regulators of food intake and energy balance, insulin and leptin, act through central regulation of neuropeptides. Similar to periphery, insulin and leptin may regulate hypothalamic proglucagon expression and thereby PGDPs. Additionally, the proglucagon gene in the periphery can be regulated by cAMP-dependent pathways; however, cAMP-mediated regulation of hypothalamic proglucagon remains unknown. The lack of knowledge in this area is due to inaccessibility to the hypothalamic proglucagon neurons due to the complex architecture of the hypothalamus. Using mHypoE-39 and mHypoE-20/2 cell lines, derived from embryonic mouse hypothalamus, and mHypoA-2/10 neuronal cells, derived from adult mouse hypothalamus, we studied the mechanisms involved in the direct regulation of proglucagon gene expression by insulin, leptin and cAMP. We determined that insulin, in an Akt-dependent manner, significantly induced proglucagon mRNA expression by 70% in adult mHypoA-2/10 cells, while significantly suppressing it by 45% in embryonic mHypoE-39 cells. We further found that leptin also regulated proglucagon mRNA expression in these cell models via the Jak/Stat pathway. Leptin caused an initial increase by 66% and 43% at 1 h followed by a decrease by 45% and 34% at 12 h time points in mHypoA-2/10 and mHypoE-39 cells, respectively. Further, cAMP activation by forskolin upregulated proglucagon expression by 87% at 4h time point in mHypoE-39 cells, and it increased proglucagon gene expression through the Epac activation in mHypoE-20/2 cells. Transient transfection analysis determined that specific proglucagon 5[apos] flanking gene regions were regulated by cAMP in the embryonic cell lines, whereas an RNA stability assay demonstrated that proglucagon mRNA stability in the adult cell line was increased by leptin and insulin at 1 h and 4 h post-treatment, respectively. These findings suggest that insulin, leptin and cAMP act directly on specific hypothalamic neurons to regulate proglucagon gene expression. The PGDPs are key regulators of feeding behavior, thus a better understanding of the regulation of hypothalamic proglucagon neurons is important to further expand our current knowledge of feeding circuits.[br][br]Sources of Research Support: CIHR, CDA, BBDC, CFI, CRC, and NSERC.[br][br]Nothing to Disclose: PSD, FDE, DMI, DDB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 575 177 1163 SUN-106 PO27-01 Sunday 958 2012


957 ENDO12L_SUN-107 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Nesfatin-1 Modulates the Phosphorylation Levels of cAMP Response Element Binding Protein for Intracellular Signaling in the Neural and Islet Beta Cell Lines Emi Ishida, Koshi Hashimoto, Hiroyuki Shimizu, Tetsurou Satoh, Shuichi Okada, Masanobu Yamada, Masatomo Mori Gunma University Graduate School of Medicine, Maebashi, Japan; Kiryu University, Kiryu, Japan Nesfatin-1 is a novel anorexic peptide, which reduces food intake in rodents by intraventricular and intraperitoneal administration. An antagonist of Melanocortin 3/4 receptor, SHU9119, diminishes the anorexic effect of nesfatin-1 (1). The thirty amino residues in the middle potion of nesfatin-1 (referred to M30) exert a comparable anorexic effect as full-length nesfatin-1 (2). It has been demonstrated that nesfatin-1 evoked the calcium influx into the neurons of rat[apos]s hypothalamus (3) and induced the insulin secretion of islets (4), however, the molecular mechanism of intracellular signaling via nesfatin-1 is yet to be revealed. In the current study, we explored several cell lines responsible to nesfatin-1 and elucidated signal transduction of nesfatin-1. For this purpose, we introduced reporter vectors, which contain various kinds of response elements into several cell lines and performed reporter assays with nesfatin-1 and M30. As a result, we found that nesfatin-1 as well as M30 increased cAMP response element (CRE) reporter activity in mouse neuroblastoma cell lines (NB41A3). SHU9119 aborted the promoter activity and mutant M30, which exerts no anorexic effect [italic]in vivo[/italic] did not induce the CRE reporter activity in NB41A3 cells. Western blotting analysis revealed that nesfatin-1 and M30 increased the phosphorylation level of CRE binding protein (CREB), whereas, unexpectedly, they did not raise the intracellular cAMP levels. We also found that a MAP-kinase inhibitor and an L-type Ca channel blocker abolished the CREB phosphorylation induced by M30. On the other hand, nesfatin-1 decreased phosphorylated CREB in some islet beta cell lines, suggesting that in the beta cells, nesfatin-1 employs a distinct signaling pathway from that of neural cells. Furthermore, we performed radio-receptor assay with [sup]125[/sup]I-nesfatin-1 and demonstrated a saturable binding to the membrane fractions of mouse hypothalamus and NB41A3 cells, with Kd value 0.79nM and 0.17nM, respectively. The data suggested that a nesfatin-1 specific receptor should exist on mouse hypothalamus and cell surface of NB41A3 cells. Taken together, we concluded that nesfatin-1, via its receptor, modulates the phosphorylation levels of CREB for intracellular signaling in the neural and islet beta cell lines.[br][br](1) Oh-I S et al., Nature 2006; 443:709. (2) Shimizu H et al., Endocrinology 2009; 150:662. (3) Brailoiu GC et al., Endocrinology 2007; 148:5088. (4) Gonzalez R et al., J Endocrinol 2011; 208:R9.[br][br]Nothing to Disclose: EI, KH, HS, TS, SO, MY, MM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1072 177 1164 SUN-107 PO27-01 Sunday 959 2012


958 ENDO12L_SUN-108 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) An Early Blockade of Leptin Predisposes Rats to Overweight, Leptin- and Insulin-Resistance Charlotte Benoit, Hassina Ould-Hamouda, Arieh Gertler, Laurence Amar, Mohammed Taouis University of Paris-Sud, UMR 8195, Orsay, France; CNRS, Centre de Neurosciences Paris-Sud UMR8195, Orsay, France; The Hebrew University of Jerusalem, Rehovot, Israel Leptin, the product of the ob gene, regulates energy homeostasis by increasing metabolic rate and inhibiting food intake at the hypothalamic level. Through its long isoform receptor OB-Rb, leptin stimulates anorexigenic and inhibits orexigenic neurons of the arcuate nucleus (Arc) that project to the paraventricular nucleus (PVN) to adjust energy homeostasis. However, early in life, leptin effects are independently regulated of body weight and fat size. In newborn rodents, a pick of leptin appeared during the second week of life which is not related to the regulation of food intake. At this stage, leptin regulates hypothalamic neuropeptides expression. The blockade of leptin action early in life has long term consequences on energy homeostasis. However, the underlying mechanisms are still not clearly established. The present study aimed to analyze the long term effect of an early postnatal leptin blockade, using a pegylated rat leptin antagonist (pRLA), on insulin/leptin sensitivity and metabolic/endocrine parameters at the hypothalamic level. We show that a daily injection of leptin pRLA from d2 to d13 predisposes rats to overweight under chow or high fat diet at the adulthood as compared to control rats. Interestingly, pRLA treatment has promoted, at the adulthood, the onset of leptin- and insulin-resistance in both hypothalamus and liver. Furthermore, pRLA treatment modified the mRNA expression of genes implicated in energy homeostasis control as UCP2-3, AdipoR1-R2 in liver and skeletal muscle, the impairment of the xpression of these genes also known to be linked to insulin resistance and type-2 diabetes. In conclusion, the present paper highlights the important role of leptin early life, leptin blockade at this stage deeply affected metabolic/endocrine parameters and insulin/leptin responsiveness at the adulthood.[br][br]Nothing to Disclose: CB, HO-H, AG, LA, MT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1581 177 1165 SUN-108 PO27-01 Sunday 960 2012


959 ENDO12L_SUN-109 POSTER SESSION: Central Regulation of Body Weight (1:30 PM-3:30 PM) Danshen Extract Regulates Energy Metabolism by Activating the AMPK-SIRT1-PGC-1[alpha] Yoon Hee Cho, Cheol Ryong Ku, Hokyou Lee, Eun Jig Lee Yonsei University College of Medicine, Seoul, Republic of Korea; Northwestern University Feinberg School of Medicine, Chicago, IL Danshen, dried root of Salvia miltiorrhiza, is one kind of traditional Chinese medicine that has many effects on metabolic diseases. However, the mechanisms by which Danshen extract (DE) functions as a metabolic regulator remain largely unknown. Here we demonstrate that DE regulates energy homeostasis in brown adipocytes through activation of AMP-activated protein kinase (AMPK) and Sirt 1, resulting in mitochondrial biogenesis function. Furthermore, oral administration of DE to rodents exerts AMPK-Sirt1-PGC-1[alpha] levels in hypothalamus and contributes to body weight. AMPK phosphorylation and Sirt1 activation levels were increased by DE treatment in brown adipocytes as well as in hypothalamus, resulting activation of its down stream target, peroxisome proliferator-activated receptor-[gamma]coactivator-1[alpha] (PGC-1[alpha]). Activation of AMPK and Sirt1by DE in brown adipocytes enhanced mitochondrial biogenesis by induction of key metabolic regulator, PGC-1 [alpha]. The activation of these key metabolic sensors results in enhancement of uncoupling protein 1(UCP-1), known as thermogenin, accounting for the beneficial metabolic effects of DE. These results indicate that DE regulates energy expenditure through an AMPK-Sirt1-PGC-1 [alpha]-dependent mechanism in adipocytes and hypothalamus.[br][br](1) Shaun F et al., Exp Physiol 93.7 pp 773-797. (2) Huiyun Liang et al., Adv Physiol Educ 30:145-151, 2006.[br][br]Sources of Research Support: This work was supported by a grant (A085136) from the Korea Healthcare Technology R[amp]D Project, Ministry for Health [amp] Welfare Affairs, Republic of Korea.[br][br]Nothing to Disclose: YC, CRK, HL, EJL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 883 177 1166 SUN-109 PO27-01 Sunday 961 2012


960 ENDO12L_SUN-110 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) Rescue Islet Transplantation after Pancreas Graft Loss in Recipients of Simultaneous Pancreas-Kidney Transplantation Roger Lehmann, Philippe A Gerber, Richard A Zuellig, Giatgen A Spinas University Hospital, Zurich, Switzerland Simultaneous transplantation of the pancreas-kidney (SPK) or islet-kidney (SIK) are established treatment options for patients with type 1 diabetes and renal failure. Partial or total pancreas or islet graft failure with maintained renal function is a known complication with subsequent recurrence of wide blood glucose fluctuations and hypoglycemic episodes. In the case of SPK, retransplantation of the pancreas would be an option and islet transplantation (ITx) an alternative with advantages due to the minimal invasive nature and minimal side effects of the procedure, in particular if a minimal C-peptide production is still present.[br]We retrospectively analyzed the outcome of 7 patients who were treated with ITx after partial or complete graft loss of the pancreas after SPK due to different reasons (vascular occlusion, chronic and acute rejection, non-immunogenic pancreatitis).[br]All patients had brittle diabetes (mean diabetes duration before pancreas transplantation 28.9[plusmn]12.1 yrs, age 41.2[plusmn]9.5 yrs). Mean time between SPK and ITx was 10.1[plusmn]9.2 yrs. Islet transplantation was performed once in 5 patients, twice in one patient and 4 times (over a 6 yr period) in one patient, with a mean of 4[apos]203[plusmn]2[apos]271 islet number/kg/transplantation and 5[apos]347[plusmn]1[apos]551 IEQ/kg/transplantation. After SPK mean HbA1c dropped from 8.7[plusmn]1.9 % to 6.8[plusmn]1.0 %, but increased to 7.5[plusmn]0.7 % after graft failure of the pancreas (last value before islet transplantation) with an increasing number of disabling hypoglycemias. After ITx HbA1c was reduced to 6.2[plusmn]1.0 % after a mean follow-up of 41[plusmn]28 months after islet transplantation.[br]The insulin dosage could be reduced from 0.56[plusmn]0.18 IE/kg to 0.19[plusmn]0.13 IE/kg. No severe complications occurred after ITx, and there were no severe (grade II/III) episodes of hypoglycemia observed after ITx.[br]We show here that islet transplantation after total or partial pancreas graft loss in SPK is a safe and efficient method to restore glycemic control in patients with type 1 diabetes mellitus.[br][br]Nothing to Disclose: RL, PAG, RAZ, GAS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 25 178 1167 SUN-110 PO13-02 Sunday 962 2012


961 ENDO12L_SUN-111 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) Selective Uncoupling of Leptin[apos]s Metabolic and Immune Functions Jan Tavernier, Lennart Zabeau, Dirk Elewaut, Sylvie Seeuws, Brian Oldfield, Justin Rubio, Cathy Jensen VIB - UGent, Ghent, Belgium; Ghent University Hospital, Ghent, Belgium; Monash University, Melbourne, Australia; Howard Florey Institute, Melbourne, Australia The adipocyte-derived cytokine leptin acts as a metabolic switch, connecting the body[apos]s metabolism to high energy consuming processes such as reproduction and immune responses. Db/db mice lack a functional leptin receptor (LR) and display a complex syndrome with morbid obesity, infertility and immune dysfunctions. We obtained both genetic and biochemical evidence that leptin[apos]s metabolic and immune functions can be uncoupled at the LR level.[br](i) We characterised a novel morbidly obese mouse strain with an autosomal recessive monogenetic trait. Homozygous mutant FATT mice carry a spontaneous splice mutation leading to loss of the Ig-like domain (IGD) in the extracellular part of the LR. They are hyperphagic and obese but, in contrast with db/db mice, display only minimal lymphoid tissue changes.[br](ii) We generated a panel of neutralizing nanobodies targeting the LR. We identified three classes of neutralizing nanobodies targeting the CRH2, IGD or FNIII subdomains of the LR ectodomain. Only nanobodies directed against CRH2 inhibited leptin binding, showing that LR signalling can be blocked without affecting ligand binding. Treatment of healthy mice with a neutralising nanobody targeting the IGD induced weight gain and hyperinsulinaemia, but completely failed in the treatment of experimentally induced arthritis, encephalomyelitis and hepatitis. This is again in stark contrast with db/db mice that are highly resistant to such experimentally induced autoimmune diseases.[br](iii) A leptin antagonist that fails to interact with IGD paradoxically mimicked the effect of leptin on cell lines and on primary immune cells. LRs lacking IGD fail to activate JAK/STAT signalling but can activate the ERK1/2 pathway.[br]Together, our data provide the first evidence that metabolic and immune activities of leptin can be uncoupled at the receptor level.[br][br]Sources of Research Support: Fund for Scientific Research--Flanders; Ghent University; Interuniversity Attraction Poles (Belgium); National Health and Medical Research Council (Australia).[br][br]Nothing to Disclose: JT, LZ, DE, SS, BO, JR, CJ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1056 178 1168 SUN-111 PO13-02 Sunday 963 2012


962 ENDO12L_SUN-112 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) Leptin Is Required for Activated T-Cell Glucose Metabolism and Function Nancie J MacIver, Donte C Saucillo, Jeffrey C Rathmell Duke University Medical Center, Durham, NC; Duke University Medical Center, Durham, NC Leptin is a protein hormone secreted primarily by adipocytes and best known for its ability to regulate appetite and weight. However, leptin also has an important role in immune function. Leptin-deficient mice and humans have decreases in both total T cell and CD4+ T cell number along with abnormal T cell function, making them more susceptible to intracellular infections and atopic disease. Administration of recombinant leptin protein reverses both the metabolic defects and immune abnormalities.[br]The mechanisms by which leptin regulates T cell number and function are poorly understood. Activated T cells generate energy in large part by upregulating aerobic glycolysis, and work from our laboratory and others has shown that activated T cells require glucose to survive, proliferate, and produce the protein products required for immune cell function. Given that leptin is a known metabolic regulator and is required for normal T cell number and function, we hypothesized that leptin would enhance T cell glucose metabolism.[br]We found that leptin increases cellular proliferation and inflammatory cytokine (IL-2 and IFN-[gamma]) production in activated peripheral T cells. We also demonstrated that leptin is required to maintain normal glucose metabolism in activated peripheral T cells, via upregulation of Glut1 expression. Bone marrow transplant studies confirmed that leptin[apos]s effects on T cell function and metabolism were not mediated by cell extrinsic factors. While regulatory T cells (Treg) were increased in number in leptin deficiency, they were less glycolytic in the absence of leptin. Finally, leptin treatment was able to rescue the defective glucose metabolism observed in activated peripheral T cells from fasted animals.[br]Altogether, our findings suggest that leptin[apos]s role in regulating immune cell function may be mediated by its effects on cellular glucose metabolism. The results of these studies provide new insight into the mechanisms by which leptin regulates T cell number and function, and may yield new approaches to the pathogenesis and treatment of immune dysfunction in nutritional disorders.[br][br]Sources of Research Support: NIH K08 DK087944, Pediatric Endocrine Society.[br][br]Nothing to Disclose: NJM, DCS, JCR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1023 178 1169 SUN-112 PO13-02 Sunday 964 2012


963 ENDO12L_SUN-113 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) Repeat Meal of Paleolithic Diet Composition Causes Postprandial Declines in Gastric Inhibitory Polypeptide and Insulin Independently of Exercise Po-Ju Lin, Katarina Borer University of Michigan, Ann Arbor, MI Insulin sensitivity and glucose tolerance respond to both diet composition and exercise. A better blood glucose control on Paleolithic diet (PD), containing 22[ndash]40% carbohydrate, 19[ndash]35% protein, and 28[ndash]58% fat compared to Western diet (WD) (1) could be mediated by incretin gastric inhibitory polypeptide (GIP). Possible role of GIP in exercise associated changes in insulin response also was suggested by parallel changes in postprandial insulin and GIP after hypocaloric exercise (2). [bold]PURPOSE:[/bold] To determine whether GIP is responsible for lower postprandial insulin response to PD or to exercise when two PD meals follow two periods of exercise. [bold]METHODS:[/bold] Twenty-two healthy postmenopausal women participated in 1 of the 4 conditions: S or X trials with PD (PS or PX, n=4 each) or WD (WS, n=6; WX, n=8) to compare diet or exercise effects on postprandial plasma glucose (G), insulin (I), insulin to glucose ratio (I/G) and GIP concentrations. The PD contained 30% carbohydrate (CHO), 25% protein (PRO), and 45% FAT while WD contained 60% CHO, 15% PRO, and 25% FAT. The meals were provided at 10 and 17h. During X trials, subjects treadmill walked at 45% VO[sub]2max [/sub]between 7-9 h and 14-16 h. Blood was collected hourly for plasma G, I, and GIP measurements. Areas under the curve (AUC) were calculated for G, I, I/G, and GIP during the two 7-h postprandial periods. Data were analyzed with mixed model ANOVA. [bold]RESULTS:[/bold] Each of the two 803-820 kcal meals provided half of the daily energy needs. Energy expenditure during each exercise bout was 408 kcal. Postprandial G, I, I/G, or GIP after meal 1 were unaffected by diet or exercise. However, after the second meal, postprandial I, and GIP, but not G, were significantly lower to PD compared to WD meal (34% lower I AUC ,F=6.85, p=0.02, 28% lower I/G AUC,F=3.72, p=0.07, and 25% lower GIP AUC, F=5.16, p=0.04), and the effect was independent of exercise. Exercise before meal 2 increased G AUC by 9% (F=4.62, p=0.05) independently of diet composition. [bold]CONCLUSION:[/bold] Repeated consumption of PD, but not WD meal, causes parallel reduction of both I and GIP responses and is unaffected by pre-meal exercise. Second bout of exercise, on the other hand, increased postprandial glucose response regardless of diet composition. We report a novel effect of diet composition and exercise during second iteration of the two behaviors.[br][br](1)Cordain L et al., Eur J Clin Nutr 2002; 56S: S42. (2)Kelly KR et al.,Am J Physiol Endocrinol Metab 2009; 296:E1269.[br][br]Sources of Research Support: NIDDK grant R15 DK082800 and Blue Cross Blue Shield student award.[br][br]Nothing to Disclose: P-JL, KB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2050 178 1170 SUN-113 PO13-02 Sunday 965 2012


964 ENDO12L_SUN-114 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) The Effects of Short-Term Energy Imbalance on Appetite and Hunger-Related Hormones Elizabeth A Thomas, Jamie L Bechtell, Elizabeth H Kealey, Daniel H Bessesen, Marc-Andre Cornier University of Colorado School of Medicine, Aurora, CO Short-term imbalances in energy intake are common and are likely to play a role in long term weight changes. This study was designed to look at the effects of short-term over- and underfeeding on appetite ratings and hormones. 58 individuals were recruited as either Obese Prone (OP) or Obese Resistant (OR) based on self-identification, BMI, personal and family weight history. 29 OP (14 M, 15 W) and 29 OR (15 M, 14 W) subjects were studied during 3 conditions, Eucaloric (EU), Overfed (OF), and Underfed (UF), in a randomized crossover design. Each study period included a 3 day run-in diet, 1 day controlled feeding, and then a test day. The controlled feeding diet provided basal energy needs for EU and 40% above and below basal energy needs for OF and UF, respectively. Appetite measures (hunger and satiety) and blood sampling for ghrelin, PYY, insulin and leptin were performed every 30 minutes for 3 hours after a breakfast shake on the 5[sup]th[/sup] day, and the area under the curve (AUC) for each was calculated. Compared to EU, OF resulted in decreased hunger AUC and increased satiety AUC, while UF resulted in increased hunger AUC and decreased satiety AUC (p[lt]0.001 for all conditions). These changes did not differ by group and were accounted for by the fasting difference by condition. Compared to EU, OF resulted in increased leptin AUC, increased insulin AUC, and decreased ghrelin AUC (p[lt]0.001 for all comparisons) but no effect on PYY AUC. In contrast, UF as compared to EU resulted in decreased insulin AUC, increased ghrelin AUC and decreased PYY ([lt]0.001 for all comparisons) but no effect on leptin. Differences in leptin were accounted for by fasting differences. Overall, OP had lower ghrelin and higher insulin and leptin, although the higher leptin was accounted for by fat mass. There were no correlations between hormones and appetite ratings overall or in the OP, but in the OR there was a correlation between hunger and leptin (r=-0.22, p=0.04) and between changes in hunger and ghrelin (r=0.23, p=0.04). In summary, for appetite ratings, fasting values appear to drive the difference in the AUC for OF and UF. In contrast, meal-related hormones do not appear to be significantly affected by one day of energy imbalance, but change as a result of the caloric content of a meal. Thus, it appears that the appetitive response is dependent upon the previous day[apos]s caloric intake, whereas the hormonal response to overfeeding is related to the acute effects of a meal.[br][br]Nothing to Disclose: EAT, JLB, EHK, DHB, M-AC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1409 178 1171 SUN-114 PO13-02 Sunday 966 2012


965 ENDO12L_SUN-115 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) A Novel Technique for Real-Time Monitoring of Hormonal Effect Illustrated with Glucose-Dependent Insulinotropic Polypeptide (GIP) Rajesh A Pareta, Justin M Saul, Timothy E Kute, Mark Furth, Emmanuel C Opara Regenerative Medicine, Winston-Salem, NC; Pathology-Tumor Biology, Winston-Salem, NC To study the role of GIP in the pathogenesis of obesity and type 2 diabetes induced by over-nutrition, we sought to develop a simple, noninvasive bioassay to determine the biological activity of GIP based on cellular responses to interaction with its receptor, GIPR. For this purpose we utilized electrical impedance-based measurements made with the xCELLigence System (Roche Applied Science). This label-free method enables real-time monitoring of cell adhesion, morphological changes, toxic responses, and proliferation. We compared responses to GIP of two receptor-positive cell types, namely, human vascular endothelial cells (HUVEC) and the murine insulinoma line [Beta]-TC6, with two GIPR-negative cell types, fibroblasts (FB) and smooth muscle cells (SMC). Cells were seeded overnight in 96-well plates containing gold electrodes at the bottom to measure impedance, reported as a normalized Cell Index. HUVEC without added GIP showed steadily increasing Cell Index, consistent with normal spreading and proliferation in culture. Addition of GIP markedly decreased the slope of the increase in Cell Index in a dose-responsive manner over approximately 24 hours. GIP also altered the Cell Index profile of [Beta]-TC6 cells, but had no effect on the FB and SMC controls. Live cell imaging showed that the GIPR-positive cells became rounder in response to the hormone, consistent with the observed changes in impedance. Again, as expected, the GIPR-negative cells were unresponsive. We tested the specificity of the GIP effect on positive cells by addition of a monoclonal antibody (MAB0136, Abnova). Even at modest doses (3 [micro]g/ml) the antibody completely blocked the effect of GIP, and restored the initial rate of increase in Cell Index. An isotype-matched control monoclonal antibody against an irrelevant antigen had no effect. We conclude that the biological activities of GIP and neutralizing antibodies can be quantified conveniently in real-time via impedance-based time-dependent cell response profiles using readily available cell types, without constant need for purified pancreatic islets. This technology should be applicable to other hormone systems.[br][br]Nothing to Disclose: RAP, JMS, TEK, MF, ECO 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 833 178 1172 SUN-115 PO13-02 Sunday 967 2012


966 ENDO12L_SUN-116 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) The Effect of Prolonged Aerobic Exercise on Serum Adipokine Levels during a 180-km Ultra Endurance Race Kostas B Markou, Nikolaos D Roupas, Irene Mamali, Spiros Maragkos, Athanasios Tsekouras, Lydia Leonidou, Anastasia K Armeni, George K Markantes, Neoklis A Georgopoulos University of Patras, Patras, Greece; University of Patras, Patras, Greece; University of Patras, Patras, Greece; University of Patras, Patras, Greece The aim of the present study was to evaluate the effect of prolonged strenuous aerobic exercise and significant energy deficit on serum leptin, adiponectin, visfatin and resistin levels in ultra-endurance runners during a 180Km running race. Moreover, an assessment of stress response and glucoregulatory adjustments was attempted evaluating serum cortisol and insulin levels and their association with adipose tissue secretion.[br]The study included 17 highly trained ultra-endurance male athletes (mean age 51.29[plusmn]6.84 years and body mass index (BMI) 23.51[plusmn]1.90), participating in the 5th Olympian Race held in Greece on 28-29 May 2010. The runners covered a distance of 180 Km within an average time of 23 hours. Anthropometric features were assessed and training history was recorded. Serum leptin, adiponectin, visfatin, resistin, cortisol and insulin levels were measured in the morning of the race (baseline values), at the finish line (post-exercise) and approximately 16 hours after the end of the race (recovery period).[br]The estimated energy cost (caloric intake-energy expenditure) during the running race was approximately 5000 Kcal. All hormonal values of the post exercise and recovery status were corrected for plasma volume changes. Body weight and body fat mass were significantly reduced (p[lt]0.001 and p[lt]0.05 respectively) after the running race. Serum leptin levels were reduced post exercise (p[lt]0.001), increased during recovery period (p[lt]0.001), but failed to reach baseline levels (p[lt]0.001). Serum resistin levels were elevated after the race (p[lt]0.001), declined during recovery period (p[lt]0.05), but remained significantly higher compared with pre-exercise levels (p[lt]0.05). Serum cortisol levels were elevated (p[lt]0.001) and insulin levels were reduced post-exercise (p[lt]0.05), but both hormonal levels were fully restored during the recovery period (p[lt]0.001 and p[lt]0.05 respectively). On the other hand, no significant changes were documented for serum adiponectin and visfatin levels, neither during the race nor during recovery. Moreover, stepwise regression analysis underscored the greater impact of insulin levels on leptin variation, compared to body fat mass.[br]In conclusion, prolonged, strenuous aerobic exercise and negative energy balance increase serum resistin and reduce serum leptin levels, with no effect on serum adiponectin and visfatin levels. Moreover, in conditions of acute energy deficit, insulin mediates the regulatory role of leptin in energy homeostasis.[br][br]Nothing to Disclose: KBM, NDR, IM, SM, AT, LL, AKA, GKM, NAG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1565 178 1173 SUN-116 PO13-02 Sunday 968 2012


967 ENDO12L_SUN-117 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) The Influence of Intensive Physical Training on Salivary Adipokine Levels in Elite Rhythmic Gymnasts Kostas B Markou, Nikolaos D Roupas, Irene Mamali, Anastasia K Armeni, George K Markantes, Anastasia Theodoropoulou, Theodoros K Alexandrides, Michel Leglise, Neoklis A Georgopoulos University of Patras, Patras, Greece; University of Patras, Patras, Greece; International Federation of Gymnastics, Lausanne, Switzerland Exercise challenges homeostasis and establishes a new dynamic equilibrium. Elite Rhythmic Gymnasts (RG[apos]s) begin exercise at an early age, undergo physical and psychological stress and adopt negative energy balance to retain a lean physique.[br]The aim of the present study was to evaluate the effect of negative energy balance, acute and chronic physical training on salivary adiponectin, resistin and visfatin levels and their interaction with salivary cortisol and insulin levels in elite RG[apos]s. This study is unique in character, as all variables were assessed on the field of competition.[br]The study included 51 elite RG[apos]s participating in the top level tournament [ldquo]Kalamata 2010 World Cup[rdquo] in Kalamata, Greece on April 2010. 24 healthy sedentary pubertal girls were used as controls. Anthropometric values were assessed; baseline and post exercise salivary cortisol, insulin, adiponectin, resistin and visfatin levels were measured. Salivary adipokine levels were measured using commercially available enzyme immunoassay kits for serum determinations with minor modifications, as we reported in a recent study (1).[br]Comparisons regarding hormonal features between RG[apos]s and controls were adjusted for age, BMI and body fat percentage. At baseline, a significant inverse correlation was documented between salivary insulin and adiponectin levels (r=-0.316, p[lt]0.05), after controlling for the effect of age and BMI. Salivary adiponectin levels were higher (p[lt]0.001) and visfatin lower (p[lt]0.05) in RG[apos]s compared with controls, while no significant changes were observed regarding salivary cortisol, insulin and resistin levels. In elite RG[apos]s short term intensive anaerobic exercise led to increased salivary insulin levels (p[lt]0.001), reduced salivary adiponectin (p[lt]0.001) and visfatin levels (p[lt]0.05) and no changes in salivary resistin levels. Moreover, diurnal variation of salivary cortisol was lost.[br]In conclusion, chronic intensive physical training and negative energy balance up-regulate salivary adiponectin and down-regulate salivary visfatin levels, while acute glucoregulatory response to short term intensive anaerobic exercise down-regulates salivary adiponectin and visfatin levels.[br][br](1) Mamali I et al., Peptides 2011;33:120-4.[br][br]Nothing to Disclose: KBM, NDR, IM, AKA, GKM, AT, TKA, ML, NAG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1507 178 1174 SUN-117 PO13-02 Sunday 969 2012


968 ENDO12L_SUN-118 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) Anti-Inflammatory Effects of ST-26 Extracts on Atopic Dermatitis [italic]In Vitro[/italic] and [italic]In Vivo[/italic] Hye-Sun Lim, Chang-Seob Seo, Jun Kyung Lee, Hyekyung Ha, Hoyoung Lee, Hyeun Kyoo Shin Korea Institute of Oriental Medicine, Daejeon, Korea; Hanpoong Pharm [amp] Foods Co Ltd, Jeonju, Korea The seed of ST-26 (family Zingiberaceae) has long been used in traditional Chinese medicine in the treatment of inflammatory and digestive diseases. However, the atopic dermatitis (AD) of ST-26 extract has not been fully verified using scientific tools. We investigated the anti-inflammatory effect of ST-26 extract on AD [italic]in vitro[/italic] and [italic]in vivo[/italic]. The effects of ST-26 extract on the production of Nitric oxide (NO) and Prostaglandin E[sub]2[/sub] (PGE[sub]2[/sub]) in Lipopolysaccharide (LPS) stimulated RAW 264.7 macrophage and the production of thymus-and-activation-regulated chemokine (TARC/CCL17) in tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) stimulated HaCaT keratinocyte was investigated using ELISA. In the [italic]in vivo[/italic] experiment, the effects of ST-26 extract on mite antigen-treated NC/Nga mice was evaluated by measuring clinical symptoms, swelling of the skin on the back and ears, and plasma level of immunoglobulin E (IgE) and histamine. The production of NO, PGE[sub]2 [/sub]from LPS-stimulated RAW 264.7 macrophage and production of TARC from TNF-[alpha]-and IFN-[gamma]-stimulated HaCaT keratinocyte was suppressed by ST-26 extract treatment in dose-dependent manner. ST-26 extract treatment of NC/Nga mice reduced the histological manifestations of AD-like skin lesions, reduced skin dermatitis scores and decreased the levels of plasma IgE and histamine. These results suggest that ST-26 extract the development of atopic dermatitis-like skin lesions in NC/Nga mice by suppressing the inflammatory response.[br][br]Nothing to Disclose: H-SL, C-SS, JKL, HH, HL, HKS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1379 178 1175 SUN-118 PO13-02 Sunday 970 2012


969 ENDO12L_SUN-119 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) Duration of Fasting and Inhibition by Insulin May Contribute to the [italic]In Vivo[/italic] Reduction of Acyl-Ghrelin Release during a Short-Term Fast in Healthy Young Men Leon S Farhy, Ralf Nass, Jianhua Liu, Suzan Pezzoli, Bruce Gaylinn, Michael O Thorner University of Virginia, Charlottesville, VA Ghrelin is a 28-amino acid peptide released from the gut that regulates GH, appetite and possibly glucose metabolism. Acylation of serine[sup]3[/sup] with octanoate is necessary for its GH releasing activity and orexigenic effects. Acyl-ghrelin circulating levels increase before meals and at midnight, but decrease with long-term fasting (1). Also, in healthy individuals insulin acutely inhibits acyl-ghrelin release (2) and therefore, may contribute to its regulation. However, changes in insulin alone cannot explain the decline of ghrelin with fasting. The aim of this study was to test whether insulin inhibition (II) and the duration of fasting (DF) can explain the changes in acyl-ghrelin during a short-term fast. Seven normal young men, age (mean[plusmn]SD) 21[plusmn]2.9yr; BMI 22[plusmn]2.9kg/m[sup]2[/sup] were studied on the Clinical Research Center (CRC) of the University of Virginia. Volunteers were admitted to the CRC in the afternoon, had a standardized dinner after which food was not served for 37.5h. At 8am next morning, 26-h blood sampling every 10min for insulin and acyl-ghrelin was initiated. To address the aims of the study we used a mathematical model of acyl-ghrelin release and compared the performance of three sub-models approximating: II only (Model 1); DF only (Model 2); or DF+II (Model 3). We found that acyl-ghrelin levels declined progressively during the short-term fast and the within-subject average levels from 8 am (day 1) to 8pm were higher than the levels from 8pm to 8am (day 2) (p=0.014, paired t-test). This finding was in contrast to our previous long-term fasting data where a similar comparison was not significant (p=0.81), thereby supporting the model assumption that decline in acyl-ghrelin secretion indeed occurs during the short-term fast. Model 1 did not explain the acyl-ghrelin dynamics. In one subject, none of the models were able to describe the data (% variance explained [lt]15%). In the remaining 6 subjects, Model 2 explained 76% of the variance in the data, while Model 3 explained 84%; Model 3 provided a better description of the data in 5 out of 6 subjects as confirmed by the Akaike information criterion. These findings suggest a two-fold regulation of acyl-ghrelin secretion during short-term fasting: acyl-ghrelin is suppressed by endogenous insulin and depends negatively on the time elapsed since the last meal. The latter is possibly due to a depletion of octanoate in the lumen of the stomach that provides substrate for the enzyme ghrelin 0-acyl transferase.[br][br](1) Liu J, Prudom CE, Nass R, Pezzoli SS, Oliveri MC, Johnson ML, Veldhuis P, Gordon DA, Howard AD, Witcher DR, Geysen HM, Gaylinn BD, Thorner MO. Novel Ghrelin Assays Provide Evidence for Independent Regulation of Ghrelin Acylation and Secretion in Healthy Young Men. J Clin Endocrinol Metab. 93(5):1980-1987, 2008. (2) Nass R, Liu J, Pezzoli SS, Gaylinn BD, Farhy LS, Thorner MO 2011 Inhibition of Acyl-Ghrelin Release in Healthy Young Adults during Euglycemic Hyperinsulinemic Clamp 93th Annual Meeting of the Endocrine Society, Boston.[br][br]Sources of Research Support: R01DK082805, K23RR018770, M01RR 00847, R01DK076037.[br][br]Nothing to Disclose: LSF, RN, JL, SP, BG, MOT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 669 178 1176 SUN-119 PO13-02 Sunday 971 2012


970 ENDO12L_SUN-120 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) Antigen Specificity in Oral Tolerance: The Role of the Anti-Insulin B Lymphocyte Zachary A-F Kistka, Rachel A Henry, Jonathan M Williams, Chrys Hulbert, James W Thomas Vanderbilt University Medical Center, Nashville, TN; Vanderbilt University Medical Center, Nashville, TN; Vanderbilt University, Nashville, TN Type 1 diabetes (T1D) is an organ-specific autoimmune disease that results from pathologic loss of immune tolerance leading to the destruction of insulin-producing [beta]-cells in the pancreatic islets. B lymphocytes play an integral role in T1D as requisite antigen presenting cells (APCs) for activating autoreactive T cells. Oral tolerance is a promising disease prevention mechanism by which immune non-reactivity is promoted by antigen exposure through gut lymphoid structures. Delivery of one [beta]-cell autoantigen ([beta]-CAA), insulin, to the gut prevents T1D in nonobese diabetic (NOD) mice by stimulating T regulatory (Treg) cell differentiation. In human studies, treatment with oral insulin reveals that individuals with the highest level of insulin autoantibodies (IAA) show the most benefit in disease prevention. The function of B lymphocytes in oral tolerance induction is not known. Although IAAs are present in WT NOD mice, routine detection of anti-insulin B cells by flow cytometry is hindered by their low numbers. To circumvent this impasse, our laboratory developed lines of NOD mice expressing anti-insulin BCR transgenes that have traceable numbers of insulin-binding B cells. Preliminary data validates the detection of anti-insulin B lymphocytes in mesenteric lymph nodes and Peyer[apos]s patches. We hypothesize that exposure of anti-insulin B lymphocytes to insulin delivered in the gut facilitates oral tolerance through alteration of effector lymphocyte subpopulations and prevents T1D.[br]ELISPOT and flow cytometry will be used on NOD mice harboring anti-insulin transgenes to identify how anti-insulin B lymphocyte exposure to oral insulin a) skews Th1 versus Th2 differentiation, b) alters Th17 cell numbers, and c) increases FoxP3+ Treg numbers. Ig transgenic NOD mice which lack anti-insulin B lymphocytes will receive anti-insulin B lymphocytes (oral insulin-treated or untreated) to test the hypothesis that exposure to oral insulin alters trafficking of anti-insulin B cells to the gut associated lymphoid tissue (GALT). Because multiple [beta]-CAAs are attacked in T1D, it is important to determine if oral tolerance toward one autoantigen prevents autoaggression against other [beta]-CAAs. We will use BDC2.5 T cell transfer to determine whether oral insulin-induced tolerance is restricted to insulin, or results in global Treg-mediated suppression of effector lymphocytes targeting other [beta]-CAAs.[br][br]Nothing to Disclose: ZA-FK, RAH, JMW, CH, JWT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2203 178 1177 SUN-120 PO13-02 Sunday 972 2012


971 ENDO12L_SUN-121 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) Hyperglucagonemic Metabolic Syndrome in SHROB Rats: Impact of Treatment with a Peptidase (DPP-IV) Inhibitor or a Sulfonylurea Richard J Koletsky, Paul Ernsberger Case Western Reserve University School of Medicine, Cleveland, OH Inhibitors of dipeptidyl dipeptidase IV (DPP-IV) are effective antidiabetic agents in patients with type 2 diabetes. They act through increasing levels of incretins, gut hormones that act in the pancreas to increase insulin and decrease glucagon release. The SHROB rat is a prediabetic model of metabolic syndrome (1) with two-fold elevations in fasting glucagon levels on a background of 20-fold higher fasting insulin relative to lean SHR littermates (2). We sought to determine whether the DPP-IV inhibitor sitagliptin might be effective in reducing glucagon in the SHROB model relative to the sulfonylurea glyburide. Because insulin may inhibit glucagon release, increasing insulin levels with glyburide might lower glucagon secretion. SHROB were treated for 6 wk with vehicle, sitagliptin (30 mg/kg/d) or glyburide (1 mg/kg/d) and compared to untreated SHR controls. Glucagon was elevated in untreated SHROB compared to untreated lean SHR (445[plusmn]31 vs 249[plusmn]25 ng/mL). Glucagon was unchanged after vehicle treatment (460[plusmn]57 ng/mL) and similarly unchanged after glyburide (476[plusmn]43 ng/mL). Following sitagliptin, glucagon was lowered to 310[plusmn]52 ng/mL. Body weight, food intake and fasting glucose were all unchanged in all treatment groups over time. reduced 33% by sitagliptin while unchanged following glyburide or vehicle. Sitagliptin is effective in lowering excessive glucagon levels in a model of metabolic syndrome, such that glucagon no longer differs from lean controls. Despite increasing insulin levels, glyburide did not alter glucagon levels. Treatments to lower glucagon may have a role in treating metabolic syndrome.[br][br](1) Ernsberger P, Koletsky RJ, Friedman JE. Molecular pathology in the obese spontaneous hypertensive Koletsky rat: a model of syndrome X. Ann N Y Acad Sci 1999; 892:272-288. (2) Velliquette RA, Koletsky RJ, Ernsberger P. Plasma glucagon and free fatty acid responses to a glucose load in the obese spontaneous hypertensive rat (SHROB) model of metabolic syndrome X. Exp Biol Med (Maywood ) 2002; 227(3):164-170.[br][br]Sources of Research Support: Investigator Initiated Grant, Merck. Ophthalmology Education Worldwide, Cleveland, OH.[br][br]Nothing to Disclose: RJK, PE 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1018 178 1178 SUN-121 PO13-02 Sunday 973 2012


972 ENDO12L_SUN-122 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) Graft Function and Glucose Daily Profile Assessed with CGMS after Islet Transplantation for Type 1 Diabetes Mellitus Marie-Christine Vantyghem, Violeta Raverdi, Frederique Defrance, Anne-Sophie Balavoine, Robert Caiazzo, Christian Noel, Julie Kerr-Conte, Francois Pattou Lille University Hospital, Lille, France; Lille University Hospital, Lille, France; Lille University Hospital, Lille, France; INSERM, Lille, France Context: The influence of beta cell replacement on daily glucose profile in Type 1 diabetes mellitus (T1DM) is not firmly established.[br]Objective: To examine the influence of islet transplantation (IT) on the various component of dysglycemia in T1DM patients.[br]Design, setting and patients: Single arm open labeled study. Twenty-three consecutive patients with T1DM, 11 males and 12 females, 14 non uremic and 9 uremic with a previous kidney graft, receiving IT at a referral center from 2003 to 2007. All participants completed a 3-year follow-up.[br]Intervention: IT consisting of 2 or 3 intra-portal allogenic islet infusions over 3 months with the Edmonton immuno-suppression protocol.[br]Outcomes: Glucose daily profile was assessed during 72 hours by continuous glucose measurement (CGM) prior to transplantation, at 6 months and yearly during 3 years after transplantation. Outcomes were mean glucose (MG), glucose standard deviation (GSD), the percentage of time spent in hyperglycemia [gt]180mg/dL (HYPER), and the percentage of time spent in hypoglycemia [lt] 54mg/dL (HYPO). Graft function was estimated with [szlig] score, a previously validated index (range 0[ndash]8) based on insulin or oral treatment requirements, plasma C-peptide, blood glucose, and A1C. Analysis was per intention to treat.[br]Results: At 3 years, 19 patients had a functional graft and 10 remained insulin independent with A1c [le]6.5%. Median A1c in the whole cohort was 6.7% (interquartile range[IQR], 5.9%-7.7%) vs. 8.3% (IQR, 7.3%-9.0%) at baseline (p[lt]0.01). The four CGM outcomes were significantly improved vs. baseline (p[lt]0.01), in a close relation with graft function (p[lt]0.001). Partial function ([szlig] score 3) was sufficient to abrogate HYPO but optimal function ([szlig] score 8) was necessary to normalize MG, SD, and HYPER.[br]Conclusion: Glucose daily profile significantly improved during the 3-year period post-IT and reached normal values when optimal graft function was achieved.[br][br]1. de Kort H, de Koning EJ, Rabelink TJ, Bruijn JA, Bajema IM. Islet transplantation in type 1 diabetes. Bmj.2011;342:d217. 2. Ryan EA, Shandro T, Green K, et al. Assessment of the severity of hypoglycemia and glycemic lability in type 1 diabetic subjects undergoing islet transplantation. Diabetes. 2004;53(4):955-962. 3. Klonoff DC, Buckingham B, Christiansen JS, Montori VM, Tamborlane WV, Vigersky RA, Wolpert H; Endocrine Society.Continuous glucose monitoring: an Endocrine Society Clinical Practice Guideline. J Clin Endocrinol Metab. 2011;96(10):2968-79. 4. Toso C, Shapiro AM, Bowker S, et al. Quality of life after islet transplant: impact of the number of islet infusions and metabolic outcome. Transplantation. 2007;84(5):664-666. 5. Siegelaar SE, Holleman F, Hoekstra JB, DeVries JH. Glucose variability; does it matter? Endocr Rev. 2010;31(2):171-182.[br][br]Sources of Research Support: This study was supported by the French Ministry of Health(PHRC 2001), the European Community(Fond Europe[acute]en de Developpement Regional and FP7), the JDRF, Conseil Regional Nord Pas de Calais (IFR 114, Institut de Medecine Predictive et de Recherche The[acute]rapeutique), and Groupement Inter Hospitalier G4 (Amiens, Caen, Lille,Rouen); clinicalTrial.gov identifiers: NCT00446264; NCT01123187.[br][br]Nothing to Disclose: M-CV, VR, FD, A-SB, RC, CN, JK-C, FP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1305 178 1179 SUN-122 PO13-02 Sunday 974 2012


973 ENDO12L_SUN-123 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) Allogeneic Bone Marrow Sustains Human Islet [beta]-Cell Function by Increased Cell Numbers [italic]In Vitro[/italic] John ZQ Luo, JunPing Wang, Fang Xiong, Philippe Ravassard, Luguang Luo Roger Williams Hospital, Providence, RI; Alpart Medical School, Providence, RI; University Pierre et Marie Curie, Paris, France Human islet [beta]-cell regeneration is challenge for diabetic field. The capability of human islet self replication in adult is unclear. However, it has been reported that human islet [beta]-cells can be self replication to regenerate more [beta]-cells in suitable condition. In order to find whether adult human islet [beta]-cell can be self regeneration, we developed an intracellular anti-proinsulin staining coupled with quantitative cytometry analysis. Such approach provides a powerful tool to quantify [beta] cells from cultured islets.[br]We evaluated the human [beta] cell population in both islet-only and BM/islet cultures and found that the number of [beta] cells in islet-only culture declined significantly from 100% on day 3 to about 40% on day 25, and 20% on day 33, which is consistent with insulin release data 100% on Day 3, 42% on Day 25 and 18% on Day 33. Islets from BM coculture, on the other hand, showed an increase with time in [beta] cell population 28.15 on Day 4, 48.15 on Day 34 and 58.1 on Day 50.[br]To further identify human islet [beta]-cell self replication during islet culture, we used a lentiviral co-transduction method. Islets were transduced with two vectors one expressing cre under the control of the insulin promoter and a loxP reporter vector constitutively expressing GFP only after cre recombination. With such approach pre-existing and newly formed b cells will turn to be green.[br]We used 10[sup]7 [/sup]PFU/per ml of both vectors and found a 75% transduction yield in islets. Insulin positive cells show green, but none insulin cells stain red during culture. We found that [beta] cells number increases which indicate that adult human islet cells can generate new [beta]-cells. However, capability for [beta]-cell regeneration lost shortly after culture 5 days. However, exciting finding is a significant increase of insulin-positive cells in islets after 14 days coculture with BM, suggesting that BM sustains human islet [beta]-cell regeneration capability.[br]The data suggest that human [beta] cell in islets in vitro have the capability to re-growth but that such re-growth is not sustainable without BM support. Further studies will be needed to explore the mechanisms for [beta] cell number increase (self duplication, differentiation or regeneration).[br][br]Sources of Research Support: This publication was made possible, in part, by Research Grant Number 1-2007-180 for Dr. LuGuang Luo by JDRF, RWMC research fund for Dr. Luo, NIH COBRE Grant award Number 2P20RR018757-06 and ICR funded by NIH and JDRF to distribute human islets for this project.[br][br]Nothing to Disclose: JZQL, JW, FX, PR, LL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1552 178 1180 SUN-123 PO13-02 Sunday 975 2012


974 ENDO12L_SUN-124 POSTER SESSION: GI Regulatory Peptides, Transplantation, [amp] Immunology (1:30 PM-3:30 PM) Alpha Cell Hyperplasia and Increased Alpha Cell Proliferation in Response to Glucagon Receptor Loss of Function Does Not Depend on Signals from the Pancreatic Environment Ana Maria Robledo Reyes, Chunhua Dai, E Danielle Dean, Greg Poffenberger, Ian M McGuinness, Patricia M Vuguin, Maureen J Charron, Alvin C Powers Vanderbilt University Medical Center, Nashville, TN; Albert Einstein College of Medicine, Bronx, NY; Vamderbilt University Medical Center, Nashville, TN; VA Tennessee Valley Healthcare System, Nashville, TN Excessive glucagon action in type 2 diabetes contributes to impaired glucose homeostasis so approaches that reduce glucagon signaling are attractive therapeutic targets. Animal models with impaired or absent glucagon signaling, including glucagon receptor knockout mice (Gcgr [sup]-/-[/sup]), have improved blood glucose tolerance, but have hyperglucagonemia and [alpha] cell hyperplasia. To investigate the stimulus for the increased [alpha]-cell proliferation, we transplanted pancreatic islets from wild type (WT) mice beneath the kidney capsule of both WT and Gcgr [sup]-/-[/sup] mice and measured [alpha] cell area ([alpha] cell area/[alpha] + [beta] cell area) and [alpha] cell proliferation (Ki67 staining). We found a 5 fold increase in [alpha] cell area in WT islets 8 weeks after transplantation into Gcgr [sup]-/- [/sup]mice compared to transplantation into WT recipients [63.3% [plusmn] 5.4%, [italic]n[/italic]=3 vs 11.5% [plusmn] 0.5%, [italic]n[/italic]=3; [italic]p[/italic][lt]0.0001]. The increased [alpha] cell area was accompanied by an increase in [alpha] cell proliferation in the WT to Gcgr [sup]-/-[/sup] grafts 8 weeks after transplantation (4.4% [plusmn] 1.2%, [italic]n[/italic]= 4 vs 0.2 % [plusmn] 0.2%, [italic]n[/italic]=5; p[lt]0.006), which was similar to the [alpha] cell proliferation rate in the Gcgr [sup]-/-[/sup] endogenous pancreas (5.1% [plusmn] 1.2, [italic]n=[/italic]4). In separate experiments, we found that [alpha] cell proliferation was increased in WT to Gcgr [sup]-/- [/sup]grafts one week after transplantation. These results suggest that when glucagon signaling is interrupted, the increased [alpha] cell proliferation and hyperplasia are not dependent on the pancreatic environment, but rather on a circulating signal.[br][br]Nothing to Disclose: AMRR, CD, EDD, GP, IMM, PMV, MJC, ACP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1825 178 1181 SUN-124 PO13-02 Sunday 976 2012


975 ENDO12L_SUN-125 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Epicardial Fat Thickness Is Related to Left Ventricle Geometry in Subjects with Coronary Artery Disease Raffaella Poggioli, Brian Kim, Gianluca Iacobellis University of Miami Miller School of Medicine, Miami, FL Background: Epicardial fat is thought to locally modulate the heart. We showed before that epicardial fat thickness significantly and independently relates to left ventricular mass (LVM). Whether epicardial fat can play a role in modulating LV shape is unexplored. Aim: In this study we sought to evaluate whether epicardial fat thickness correlated with different LV geometric patterns. Subjects and Methods: Echocardiographic epicardial fat thickness was measured in a total of 45 male subjects: 15 men with clinically and angiographically established coronary artery disease (CAD) and metabolic syndrome (MS) (CAD+ MS+), 15 men with MS, but no history, clinical signs of CAD and with normal coronary arteries on coronary angiography (CAD- MS+) and 15 men without MS or CAD (CAD- MS-). Results: CAD+MS+ subjects had significantly higher epicardial fat thickness, LV mass (LVM), LVMh2.7 and relative wall thickness (RWT) when compared to CAD- MS+ and CAD-MS- subjects, p[lt]0.001 for all. CAD+MS+ subjects had higher both concentric and eccentric LV hypertrophy (LVH) patterns than CAD-MS+ and CAD-MS-, p[lt]0.01 for both. Epicardial fat thickness was significantly (p[lt]0.05) higher in CAD+ MS+ subjects with concentric LVH (14.5[plusmn]2 mm) and eccentric LVH (13.3[plusmn]2 mm) than in those with LV remodeling (11.5[plusmn]2 mm). Epicardial fat thickness was the best correlate of LVM, (R2=0.36, p[lt]0.05) in CAD+MS+ subjects. Conclusions: Although data are preliminary and obtain only in a male population, our observation suggests a relationship of epicardial fat thickness with LV geometry. Excessive epicardial fat can mechanically affect the heart and contribute to the development of abnormal geometry. Given that different LV geometric patterns account for different cardiovascular risk, these data, if confirmed in a larger population, can be of great interest.[br][br]Nothing to Disclose: RP, BK, GI 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1577 179 1182 SUN-125 PO28-03 Sunday 977 2012


976 ENDO12L_SUN-126 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Dietary Induced Weight Loss in Patients with Poorly Controlled Diabetes Is Associated with Rise in Adiponectin and Improved Glycemic Control Elizabeth Beale, WeiAn Lee, Martha Walker, Kati Konersman, Anne Peters, Viorica Ionut University of Southern California, Los Angeles, CA; University of Southern California, Los Angeles, CA; Cedars Sinai Medical Center, Los Angeles, CA; St Mary[apos]s Medical Center, San Francisco, CA [bold]Background:[/bold] Adiponectin regulates insulin sensitivity and has antiatherosclerotic and anti-inflammatory properties and attempts are being made to develop it as a therapeutic agent for patients with type2 diabetes (T2D) and obesity(1). Weight loss raises adiponectin levels. Insulin therapy can lead to increase in body weight and further insulin resistance, and elusive glycemic control. Low-income Hispanic patients have a particular need for novel, effective therapeutic strategies.[br][bold]Aim:[/bold] To evaluate the effect of weight loss induced through caloric restriction on glycemic control, insulin requirements and adiponectin levels in low-income Hispanic patients withT2D[br][bold]Design:[/bold] Pilot, prospective, randomized, controlled trial.[br][bold]Participants:[/bold] Low-income, obese, Hispanic adults with type 2 diabetes, A1c[gt] 8% on insulin attending a specialist-led community-based staged diabetes management program.[br][bold]Method:[/bold] Participants were randomized to standard care (SC) or lifestyle (LS). LS received free [sim]1200kcal/day meal replacements for 3months(m) (HMR) and offered monthly group education sessions and advised to exercise regularly. Paired 2-tailed t-test was used for intragroup and unpaired for intragroup comparisons.[br][bold]Results:[/bold] 15 participants/group, with baseline (BL) and 3m data on 10 LS and 9 SC participants. There was no significant difference between LS vs SC (mean(sd))at BL in: age(yrs)48(8) vs 49(8); M:F: 2:8 vs 1:8; BMI(kg/m[sup]2[/sup]): 43(14)vs 37(5); duration of T2D (yrs)18(12) vs 14(9);insulin (Units/day):82(72) vs 78(59); A1c (%)9.6(1) vs 9.5%(1); or adiponectin levels:5590(3976)vs 7762(6438).[br]At 3m compared with BL, LS had lost a significant amount of weight, had improved glycemic control, reduced total daily dose of insulin and increased adiponectin levels, none of which were seen with SC.LS vs BL(p)::SC vs BLmean(sd)(p): BMI:43(14)vs41.5(13)(p[lt]0.01):: 37(5):36.6(5)(p=0.8); A1c:9.6(1):8.1(1)(p=0.02):: 9.5(1):8.7(2)(p=0.2); insulin 82(72):50(58)(p[lt]0.01):: 78(59):82(43)(p=0.6); adiponectin:5590(3976):7012(4413)(p[lt]0.01)::7762(6437): 8241(7753)(p=0.5).[br][bold]Conclusion:[/bold] Weight loss achieved through use of MR and LS modification in a group of patients with refractory T2D is associated with significant clinical benefit and a reduction in total daily insulin requirement. Adiponectin rise may mediate this benefit. This study justifies further exploration of non-pharmacologic strategies to raise adiponectin particularly in patients unwilling or unable to use insulin effectively.[br][br](1)Silva FM et al,Nutrition Reviews[reg] 2011 Vol. 69(10):599[ndash]612.[br][br]Sources of Research Support: HMR; Unrestricted Grant in kind awarded to EB.[br][br]Nothing to Disclose: EB, WL, MW, KK, AP, VI 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1818 179 1183 SUN-126 PO28-03 Sunday 978 2012


977 ENDO12L_SUN-127 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Low Glycemic Index, Low Glycemic Load Dessert Consumption Has a Favorable Effect on Weight Loss, Fat Distribution, Metabolic/Hormonal Profile and Compliance in Both Overweight/Obese Children and Adults Undergoing Nutritional Intervention Sarantis Livadas, Antonia Dastamani, Aphrodite Dikaiakos, Aimilia Mantziou, George P Chrousos University of Athens, Athens, Greece Introduction: The importance of nutritional intervention in obese subjects is crucial for cardiovascular risk reduction. However, the majority of patients following dietary intervention programs discontinue their follow-up during the first 3-6 months, mainly due to poor compliance to dietary habit modification.[br]Purpose: To investigate the effect of consumption of foods with low Glycemic Index/Glycemic Load (GI/GL), weight loss, fat distribution, metabolic/hormonal profile and the degree of compliance in overweight/obese children and adults who follow a diet plan.[br]Patients: We studied 28 teenage girls (10-14 years) with BMI [ge] 85th percentile and 37 adults (33 women, 4 men), mean age 42 years with body weight: BMI: 32kg/m2.[br]Methods: All study participants followed a hypocaloric diet (carbohydrates 45%, lipids 35%, protein 20%) and, depending on their preference, they were divided into two groups: Group A (n=14 children, 19 adults), allowing the consumption of low GI/GL desserts 4 times/wk, and Group B (n=14 children, 18 adults) allowing the consumption of one dessert once a week. Subjects[apos] compliance to diet was evaluated every two weeks and anthropometric characteristics, metabolic/hormonal profile (lipids, glucose, insulin, leptin, adiponectin, hsCRP, IL-6), were evaluated at the beginning of intervention and at 45 and 90 days of follow-up. Furthermore, in adults, fat percentage and distribution was assessed at the same time intervals.[br]Results: BMI, glucose values and insulin resistance index (HOMA-IR), significantly improved in both children and adults at the end of dietary intervention (p[lt]0.05). Moreover, in adults there was a significant improvement in lipid profile and percent fat. In children, significant improvements of insulin levels and HOMA-IR were observed in group A compared to group B. Compliance with the low GI/GL diet tended to be higher in group A.[br]Conclusions: A nutritional program offering low GI/GL desserts was effective in achieving weight loss. Furthermore, this type of nutritional approach improved metabolic/hormonal profile and increased compliance in both children and adults. Thus, consumption of low GI/GL desserts is recommended in subjects undergoing dietary intervention as it increases compliance and improves the metabolic profile.[br][br]Nothing to Disclose: SL, AD, AD, AM, GPC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 389 179 1184 SUN-127 PO28-03 Sunday 979 2012


978 ENDO12L_SUN-128 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Psyllium Supplementation Improves Parameters of the Metabolic Syndrome: A Randomized, Participant-Blinded, Placebo-Controlled, Crossover Trial Martin Isaac de Bock, Jose Derraik, David Cameron-Smith, Greg Smith, Clare Wall, Wayne Cutfield University of Auckland, Auckland, New Zealand; University of Auckland, Auckland, New Zealand; University of Auckland, Auckland, New Zealand Objective: To assess the effects of psyllium supplementation on insulin sensitivity and other parameters of the metabolic syndrome in an at risk adolescent population.[br]Design: Participant-blinded, randomized, placebo controlled, crossover trial[br]Subjects and Methods: 47 healthy adolescent males aged 15[ndash]16 years were recruited from secondary schools in lower socio-economic areas with high rates of obesity. Participants received 6 g/day of psyllium or placebo for 6 weeks, with a two-week washout before crossing over. Fasting lipid profiles, ambulatory blood pressure, auxological data, body composition, activity levels, and three-day food records were collected at baseline and after each 6-week intervention. Insulin sensitivity was measured by the Matsuda method using glucose and insulin values from an oral glucose tolerance test.[br]Results: 45 subjects completed the study, and compliance was very high: 87% of participants took [gt]80% of prescribed capsules. At baseline, 44% of subjects were overweight or obese. 28% had decreased insulin sensitivity, but none had impaired glucose tolerance. Fibre supplementation led to a 4% reduction in android to gynoid fat ratio (p=0.019), as well as a 0.12 mmol/l (6%) reduction in LDL cholesterol (p=0.042). Further, when baseline fibre intake was controlled for in the analyses, the overnight systolic dip was 1.2-fold greater with psyllium treatment (p=0.055). No associated adverse events were recorded.[br]Conclusions: Dietary supplementation with 6 g/day of psyllium over 6 weeks improves parameters of the metabolic syndrome in an at risk population of adolescent males.[br][br]Sources of Research Support: Starship Foundation, Australasian Paediatric Endocrine Group, and New Zealand Society for the Study of Diabetes.[br][br]Nothing to Disclose: MIdB, JD, DC-S, GS, CW, WC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 414 179 1185 SUN-128 PO28-03 Sunday 980 2012


979 ENDO12L_SUN-129 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Patients with Primary Pigmented Nodular Adrenocortical Disease (PPNAD) Are Less Obese Than Matched Patients with Cushing Syndrome; Is It Due to Increased PKA Activity? Edra London, Maria Nesterova, Margaret Keil, Maya Lodish, Rania Dagalakis, Stratakis A Constantine National Institute of Child Health and Human Development, National Institutes Health, Bethesda, MD Cushing syndrome (CS) is caused by hypercortisolemia initiated by adrenocortical hyperplasia and/or cortisol-producing adenomas (CPA), or ACTH-secreting tumors. Bilateral adrenocortical hyperplasia (BAH) diseases associated with CS may be caused by mutations in genes coding for proteins involved in cAMP signaling. Mutations of the PRKAR1A gene that codes for PKA regulatory I-[alpha] subunit (RI[alpha]) cause Carney complex (CNC), which is associated with PPNAD. It is not known how PRKAR1A mutations or abnormalities in PKA signaling may impact the obese phenotype associated with CS. While PKA activity has been investigated in adrenocortical tumors, it has not been explored in periadrenal adipose tissue (PAT). We studied the BMI or BMI z-score, and quantified PKA activity and PKA subunit expression in PAT in various groups of patients with CS. PAT was collected during adrenalectomy from 48 patients with CS We also studied 9 patients with aldosterone producing adenoma (APA) that served as non-CS controls for PAT studies. PKA activity assay and quantitative RT-PCR and western blot were performed for all PKA subunits. Height, weight and age data for a total of 216 patients with CS and Cushings disease (CD) were analyzed. Among adult CS patients those with PPNAD who harbored PRKAR1A mutations (mtPPNAD) had lower BMI (p[lt] 0.04) than CS without PRKAR1A mutations and were comparable to APA controls. In pediatric patients, PPNAD patients had significantly lower BMI z-score than those with CD (p= 0.01). Interestingly, in the pediatric cohort, total PKA activity in PAT was higher (p=0.007) and the ratio of basal to total PKA activity was lower (p=0.002) in mtPPNAD compared to CS patients without PRKAR1A mutations. Similar trends were present in adult patients, but they did not achieve statistical significance. PKA regulatory II-[alpha] subunit (RII[alpha]) and catalytic [alpha] subunit (C[alpha]) mRNA expression were significantly higher in mtPPNAD compared to APA PAT (p[lt]0.005 and p[lt]0.002, respectively). PKA C[alpha] mRNA expression was higher in mtPPNAD compared to CS without PRKAR1A mutations (p=0.04). These results suggest that PPNAD and mtPPNAD patients may be less obese due to increased cAMP signaling. Differences between adult and pediatric CS patients suggest that there are also age-related changes in PKA activity in PAT which may determine both visceral adiposity and clinical phenotype in patients with CS.[br][br]Nothing to Disclose: EL, MN, MK, ML, RD, SAC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2021 179 1186 SUN-129 PO28-03 Sunday 981 2012


980 ENDO12L_SUN-130 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) The Endocrine and Metabolic Characteristics of a Large Bardet-Biedl Syndrome Clinic Population Safa Mujahid, Mohammed Shahriar Bobby Huda, Elizabeth Forsythe, Jonathan Hazelhurst, Jeremy Tomlinson, Philip Beales, Paul Carroll, Barbara McGowan Guys[apos] [amp] St Thomas[apos] NHS Foundation Trust, London, UK; University of Birmingham, Birmingham, UK Background: Bardet Biedl syndrome (BBS) is a rare autosomal recessive disorder caused by ciliary dysfunction. It is characterised by obesity, rod cone dystrophy, polydactyly, hypogonadism, renal abnormalities and cognitive impairment. There are currently few data on endocrine and metabolic abnormalities in adult patients, and we examined this in a large BBS population.[br]Methods: One hundred and forty-three patients with BBS were identified from two national BBS clinics. Fasted blood samples were taken in all patients and morning pituitary profiles were measured in 123 patients. Metabolic syndrome was defined using the International Diabetes Federation (2006) criteria. Non-alcoholic fatty liver disease (NAFLD) was suggested by the presence of elevated liver enzymes. Chronic kidney disease (CKD) was staged using estimated glomerular filtration rate (eGFR). Means are represented [plusmn] standard error of the mean (SEM).[br]Results: Seventy-seven (53.8%) were male. Mean age was 31.6 [plusmn] 1.0 yrs (range 13-57 yrs) and BMI was 35.5 [plusmn] 0.7kg/m2; 93(74.4%) were obese (BMI[gt]30 kg/m2). The prevalence of metabolic syndrome was high (50.3%) and 19(13.3%) patients had type 2 diabetes. Polycystic ovarian syndrome was present in 11/66 (16.7%) females and 29(24%) BBS patients had biochemical evidence suggestive of NAFLD.[br]Ninety-five (77.2%) patients were eupituitary. Of the remaining patients 17(13.8%) had an isolated low IGF-1 (thought to be secondary to obesity), 7 had mild hyperprolactinaemia (prolactin [lt]1500mIU/l) and 7 patients had isolated low prolactin. One patient had significant hyperprolactinemia (prolactin= 6391mIU/ml) and a subsequent MRI showed pituitary hypoplasia. Twenty-four (40.7%) males were hypogonadal; this was primary in 4 patients and secondary in 16 patients. Two patients had treated nephrogenic diabetes insipidus. Most patients were euthyroid (74.8%), 8(6.7%) patients had hypothyroidism and 1 patient had hyperthyroidism. Subclinical hypothyroidism was present in 14.3%. Five (4.3%) patients had stage 5 CKD, 4(3.5%) had stage 4 CKD and 14(12.2%) stage 3 CKD. Four patients had functioning renal transplants.[br]Conclusions: This is the first study to investigate endocrinopathies associated with BBS in a large clinic population. Despite previous reports, generalised pituitary hormone dysfunction is not common but hypogonadism, when present, is often central in origin. The majority of patients are obese and the prevalence of metabolic syndrome and type 2 diabetes is high.[br][br]Nothing to Disclose: SM, MSBH, EF, JH, JT, PB, PC, BM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2101 179 1187 SUN-130 PO28-03 Sunday 982 2012


981 ENDO12L_SUN-131 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Pseudohypoparathyroidism Type 1A Patients Have Increased Insulin Resistance and May Be More Prone to Diabetes Than Similarly Obese Controls Mary A Warren, Gail R Hall, Xiongce Zhao, Emily L Germain-Lee, Lee S Weinstein, Monica C Skarulis NIDDK, National Institutes of Health, Bethesda, MD; NIDDK, National Institutes of Health, Bethesda, MD; Kennedy Krieger Institute, Baltimore, MD; Johns Hopkins University School of Medicine, Baltimore, MD; NIDDK, National Institutes of Health, Bethesda, MD [bold]Background: [/bold][br]G[sub]s[/sub][alpha] [sub][/sub]is a ubiquitously expressed G protein that mediates receptor-stimulated cAMP generation. Pseudohypoparathyroidism type 1A (PHP1A) is a monogenic obesity disorder resulting from heterozygous loss-of-function mutations inherited on the maternal G[sub]s[/sub][alpha] allele. Similarly, mice with maternal G[sub]s[/sub][alpha] mutations develop obesity due to reduced energy expenditure as well as insulin-resistant diabetes. To date, no systematic studies have been done to characterize the metabolic phenotype of PHP1A patients.[br][bold]Objective:[/bold][br]To examine the metabolic phenotype of adult PHP1A patients as compared to similarly obese patients without PHP1A.[br][bold]Methods[/bold]:[br]Subjects included PHP1A (n=8) and controls matched for gender, race/ethnicity, age ([plusmn]5 years), and percent body fat (BF%) by air displacement plethysmography ([plusmn]6%). Insulin sensitivity (SI), acute insulin response to glucose (AIRg), insulin-dependent (DI) and insulin-independent glucose disposal (SG) were derived by an intravenous glucose tolerance test. Mixed meal glucose tolerance test measured oral glucose insulin sensitivity (OGIS).[br][bold]Results[/bold]:[br]PHP1A patients included 7 females and 1 male, mean age 40 years (range 27 to 50). BF% did not differ between PHP1A and controls (39 [plusmn] 3 vs 38 [plusmn] 4%, respectively). Half of the PHP1A subjects had diabetes compared to none of the controls. HgbA1c% was significantly higher in PHP1A patients (6.13 [plusmn] 0.39 vs 5.19 [plusmn] 0.11%, p=0.038). PHP1A patients tended to have higher fasting glucose compared to controls (114 [plusmn] 14 vs. 98 [plusmn] 5 mg/dl) and had significantly lower SI (2.90 [plusmn] 0.57 vs 5.96 [plusmn] 1.02 M/mU/l), DI (616 [plusmn] 202 vs 1500 [plusmn] 457) and OGIS (384 [plusmn] 16 vs 571 [plusmn] 13 ml/min). Furthermore, SI (3.03 [plusmn] 0.77 vs 7.02 [plusmn] 1.80) and DI (871 [plusmn] 270 vs 2260 [plusmn] 681) remained significantly lower in the PHP1A patients even when the diabetic patients were removed. AIRg and SG did not differ between groups.[br][bold]Conclusions[/bold]:[br]These results indicate that adults with PHP1a are more insulin resistant and may be at greater risk for diabetes compared to similarly obese individuals. Haploinsufficiency of G[sub]s[/sub][alpha] may impact the ability of [beta]-cells to respond to incretins and may contribute to this observation. Further studies are required to determine if there is also a deficit in insulin secretion.[br][br]Nothing to Disclose: MAW, GRH, XZ, ELG-L, LSW, MCS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1748 179 1188 SUN-131 PO28-03 Sunday 983 2012


982 ENDO12L_SUN-132 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Longitudinal Multicenter Analysis on the Course of Glucose Metabolism in Obese Children Antje Korner, Susanna Wiegand, Andreas Hungele, Sabine Tuschy, Klaus Peter Otto, Dagmar laAllemand-Jander, Kurt Widhalm, Wieland Kiess, Reinhard W Holl University of Leipzig, Leipzig, Germany; Charit[eacute], Berlin, Germany; University of Ulm, Ulm, Germany; SANA Klinikum Lichtenberg, Berlin, Germany; Wilhelmstift, Hamburg, Germany; University St Gallen, St Gallen, Switzerland; University of Wien, Wien, Austria Objective: Although there is evidence of increasing prevalence of impaired glucose metabolism in obese children from smaller single cohorts, data are lacking on the progression of glucose metabolism in this patient group.[br]We aimed to assess the prevalence and the longitudinal course of impaired glucose metabolism assessed by oGTT in a large multi-center pediatric obesity registry. [br]Patients [amp] Methods: We performed an observational multicentre (n=84) longitudinal analysis on the course of glucose metabolism evaluated by oGTT in 11 156 obese children documented in the APV registry. Patients were stratified with impaired fasting glucose (IFG), impaired glucose tolerance (IGT) and type 2 diabetes (T2D) according to ADA criteria.[br]Results: A total of 12.6% of the children presented with abnormal glucose metabolism (5.99% IFG, 5.51% IGT, 1.07% T2D). BMI correlated modestly with 2h blood glucose (r=0.04, P[lt]0.001).[br]In the 1008 patients with follow-up oGTT, metabolic parameters improved and the percentage of abnormal glucose metabolism decreased from 18.7% to 14.2%. Of the children with initial IGT, 70.6% converted to normal glucose tolerance. The improvement in oGTT results was associated with, but not dependent on a reduction of BMI of -0.094[plusmn]0.42 SDS.[br]Conclusions: In summary, we provide evidence for significant improvement of oGTT parameters in obese children treated in specialized treatment centers, even though reduction in BMI was only modest.[br][br]Sources of Research Support: This study was supported by the German Ministry of Education and Research (BMBF) within the scope of the competence network [ldquo]Obesity[rdquo] LARGE consortium (to AK, SW, WK, RH) FKZ 01610824, by the competence net diabetes of the BMBF (FKZ 01GI0859), by the German Research Council (DFG) KO3512/1, by the LIFE (Leipzig Research Center for Civilization Diseases, Universit[auml]t Leipzig), funded by the European Union, by the European Regional Development Fund (ERFD) by means of the Free State of Saxony within the framework of the excellence initiative, and by the Beta-JUDO consortium funded by the European Union within the 7th framework programme.[br][br]Nothing to Disclose: AK, SW, AH, ST, KPO, Dl-J, KW, WK, RWH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2223 179 1189 SUN-132 PO28-03 Sunday 984 2012


983 ENDO12L_SUN-133 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Skeletal Muscle Cutpoint Associated with Metabolic Syndrome Risk in Middle, Older Aged Men and Women Seongsu Lee, JangWon Son, OakKee Hong, Sungrae Kim, Soon Jib Yoo The Catholic University of Korea, The Bucheon St Mary[apos]s Hospital, Bucheon, Republic of Korea Objectives: To evaluate the association between skeletal muscle mass index (SMI= skeletal muscle mass/body mass *100) cutpoint and metabolic syndrome risk among middle, older aged men and women.Methods and procedure: The Fourth Korea National Health and Nutrition Examination Surveys (KNHANESIV) in the Korean population conducted in 2008. 970 Males and 1357 females aged 40 years and older selected in the 16 administratives districts of South Korea IV were analyzed. The participants who were performed dual X-ray absorptiometry classified the metabolic syndrome by Modified National Cholesterol Education Program Adult Treatment Panel III criteria.RESULTS:At a cutoff of 38.5 percent in male aged 40 years and older, the sensitivity and specificity of prediction of metabolic syndrome were 62.3% and 69.2%, respectively,(area under the curve 0.690, p [lt] 0.001). At a cutoff value of 29.2 percent in female aged 40 years and older, the sensitivity and specificity were 58.24% and 72.2%, respectively,(area under the curve 0.696, p [lt] 0.001).[br][br](1) Lee et al., Diabetes Research and Clinical Practice 75;2007;72[ndash]80. (2) KR Fontaine et al.,International Journal of Obesity 2011;1.[br][br]Nothing to Disclose: SL, JS, OH, SK, SJY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2375 179 1190 SUN-133 PO28-03 Sunday 985 2012


984 ENDO12L_SUN-134 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Systemic Neuropeptide Y Levels Are Suppressed in the Metabolically Healthy Obese Carlo Casale, Lei Shen, Sana Malik, Rosaire Gray, Pratik Sufi, Dugal Heath, Nephtali Marina, Vidya Mohamed-Ali University College London, London, UK; Whittington Hospital, London, UK Fifty percent of a morbidly obese population are type 2 diabetic (DM), while of the remaining non-diabetic cohort, 30-40% are pathologically obese (PO), and 10-20% being metabolically healthy obese (MHO). Despite the obesity, these groups vary significantly with respect to levels of insulin resistance and their associated pathology. Elevated neuropeptide Y (NPY) could explain the different phenotypes. Caucasian subjects undergoing surgical weight-loss were studied. MHO individuals characterized as non-diabetic patients with absence of CVD, fasting insulin levels l [lt]6.5 miU/ml, fasting plasma glucose [lt]6.8 mmol/L, systolic/diastolic BP less than 140/85 mmHg. Adipokine levels were determined by ELISA in serum, adipose tissue and media from organ cultures. Adipose tissue morphology was assessed by histology and insulin resistance by HOMA-IR. Vaso-contractile function of isolated adipose tissue arterioles was determined by wire myography and angiogenesis by bioassay. Expression of angiogenic and adipogeneic genes in whole tissue was determined by PCR array. The groups did not differ by BMI. The MHO group had lower insulin levels compared to PO and DM patients (p [lt]0.001) and lower HOMA-IR (p [lt]0.001), triglycerides (p [lt]0.001) and SBP (p = 0.05). Circulating adiponectin was significantly higher in MHO individuals (p = 0.001) and in DM (p = 0.03), perhaps due to hypoglycemic therapy, compared to PO. IL-6 and MCP-1 were not different between groups. However, subcutaneous adipose tissue (SAT) leptin in PO was higher than in the MHO SAT, as a marker of adipocyte hypertrophy, as confirmed by histology in both subcutaneous and omental depots (p = [lt]0.001). Circulating NPY levels: DM 16.1(8.15-27), PO 13.1(6.6-18.6) and MHO [7.4 (3.8-12.4) pg/ml. Similar trends were apparent in adipose tissue NPY protein levels: DM [SC = 228.5(65.2-708), OM = 1255(588-1500)], PO [SC = 165.8(68.8-480.5), OM = 1048(711-1500)] and MHO group [SC = 59.4(10.2-168.1), OM = 987.6(338-1379)] pg/ml/0.5g tissue. While adipogenic genes were not different in the SAT of MHO versus PO, many angiogenic genes were up-regulated in PO SAT. NPY was also induced angiogenesis in an in vitro bioassay, while inducing transient vasoconstriction in adipose tissue micro-vessels.[br]Circulating NPY levels were singular in demonstrating a graded response between the three groups studied, and, may mediate the differences in insulin resistance and endothelial dysfunction that underlie the DM, PO and MHO phenotypes.[br][br]Sources of Research Support: UCL IMPACT Studentship.[br][br]Nothing to Disclose: CC, LS, SM, RG, PS, DH, NM, VM-A 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1907 179 1191 SUN-134 PO28-03 Sunday 986 2012


985 ENDO12L_SUN-135 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Molecular Impact of Bariatric Surgery on Human Adipose Tissue Anoopa A Koshy, Alexandria Bobe, Karla A Temple, Matthew J Brady University of Chicago Medical Center, Chicago, IL As prevalence of obesity and its associated co-morbidities have continued to rise, bariatric surgery has emerged as a front line treatment for severe obesity. In addition to promoting weight loss, the biliopancreatic diversion with duodenal switch (DS) surgery may be more effective in resolving the metabolic co-morbidities in obese patients than the more widely used Roux-en-Y gastric bypass. The goal of the studies was to determine the molecular basis by which bariatric surgery, specifically DS, might affect insulin sensitivity in human adipose tissue.[br]Subjects were recruited from the Center for Surgical Treatment of Obesity at University of Chicago and subcutaneous adipose tissue was obtained by needle biopsy from the same patient two weeks before and after bariatric surgery. Adipocytes were isolated from the fat biopsy tissue by collagenase digestion immediately following the biopsy procedure. Purified adipocytes were incubated at 37[sup][deg] [/sup]C for 10 minutes with increasing amounts of insulin, and the reaction was terminated by addition of Laemmli buffer and boiling. Samples were stored at 80[sup][deg] [/sup]C until both biopsies from each subject had been performed. Insulin sensitivity was assessed pre-and post-surgery by anti-phospho-AKT immunoblotting using the same gel for all samples from each patient. In parallel, mRNA was purified from the samples and analyzed by quantitative RT-PCR for specific proteins in the insulin signaling pathway leading to the phosphorylation of AKT: IR-[beta], IRS-1, the regulatory and catalytic subunits of PI3K and PDK1. Immunoblotting was performed to assess the regulation of these proteins before and after bariatric surgery.[br]Our data showed that DS induced an increase in insulin-stimulated AKT phosphorylation in primary human adipocytes from 3 subjects within two weeks after surgery, before any significant long-term weight loss occurred. No detectable change in the expression of the regulatory or catalytic domains of PI3K was observed with the change in insulin signaling. In contrast, preliminary results indicated there was an increase in IRS-1, as detected by qRT-PCR and confirmed by immunoblotting. These findings support the hypothesis that DS acutely improves insulin signaling at the cellular level of the adipocyte. Successful completion of this study will provide novel insights into the beneficial effects of DS, as well as potentially identify new therapeutic targets to improve insulin sensitivity independent of bariatric surgery.[br][br]Nothing to Disclose: AAK, AB, KAT, MJB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 44 179 1192 SUN-135 PO28-03 Sunday 987 2012


986 ENDO12L_SUN-136 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Obesity: An Effect of Environmental Pollutants? Catherine W McCollum, Anne Riu, Maria Bondesson, Jan-Ake Gustafsson University of Houston, Houston, TX Human health has been shown to be affected by the increasing presence of environmental pollutants and teratogens in forms of pesticides and industrial products. Concurrently, there is a growing prevalence in nationwide adulthood as well as childhood obesity, which leads to medical complications such as heart diseases, diabetes and other metabolic abnormalities. There is a strong likelihood that the obesity epidemic is tied to the increased exposure of these hazardous chemicals, some of which have been reported to be bioaccumulative. Previously reported by Riu et al. (2011) (1), tetrabromobisphenol A (TBBPA) and tetrochlorobisphenol A (TCBPA), both commonly used as flame retardants, activated human and zebrafish peroxisome proliferator-activated receptor gamma (PPAR[gamma]) in cell lines and induced adipogenesis in 3T3-L1 preadipocytes. PPAR[gamma], highly expressed in adipose tissues, promotes adipogenesis and regulates lipid metabolism. To further investigate the link between environmental pollutants and obesity, we have established zebrafish as an obesogen model. Here, we show that exposure to halogenated bisphenol A promotes lipid accumulation in zebrafish embryos. The hypothesis that increased lipid accumulation is caused by exposure to TBBPA or TCBPA is further supported by the upregulation of PPAR[gamma] downstream genes.[br][br](1) Riu et al., Environ Health Perspect 2011 Sep; 119(9):1227-32.[br][br]Nothing to Disclose: CWM, AR, MB, J-AG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 358 179 1193 SUN-136 PO28-03 Sunday 988 2012


987 ENDO12L_SUN-137 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) A 2-Year Diet Intervention Increases Systemic 11[beta]HSD1 and 5[alpha]-Reductase Activity in Obese Postmenopausal Women Andreas Pettersson, Kotryna Simonyte, Mats Ryberg, Ruth Andrew, Brian Walker, Tommy Olsson Medicine, Ume[aring], Sweden; Centre for Cardiovascular Science, Edinburgh, UK Tissue-specific glucocorticoid (GC) exposure may be key to obesity-related complications. 11[beta]-hydroxysteroid dehydrogenase 1 (11[beta]HSD1) converts cortisone into cortisol thus amplifying GC availability. In obesity, 11[beta]HSD1 activity is increased in subcutaneous adipose tissue (SAT) and decreased in liver. Short-term studies suggest that weight loss reduces GC excretion in urine, hepatic 5[alpha]-reductase activity and 11[beta]HSD1 mRNA in SAT. A low-carbohydrate diet may increase hepatic 11[beta]HSD1 activity. We investigated the effects of 2-year diet interventions on GC metabolism in obese postmenopausal women.[br]71 postmenopausal women were randomized in two groups, a modified paleolithic diet (PD; 30E% protein, 30E% fat and 40E% carbohydrates, n=37), or a diet according to the Nordic nutrition recommendations (NNR; 15E% protein, 25E% fat and 60E% carbohydrates, n=34). Food diaries and urinary nitrogen excretion were used to validate diet compliance. GC metabolites were measured using GCMS in 24-hour urine samples. 11[beta]HSD1 mRNA was analysed in SAT biopsies. Effects of diet and time were assessed with generalized estimating equations.[br]After 2 years 28 (PD) and 21 (NNR) participants remained in the study. Both groups lost weight; at 6 months the PD group had lost more weight than the NNR group (PD -9kg, NNR -5kg, diet x time effect p[lt]0.001) but this difference was not statistically significant at 24 months (PD -9kg, NNR -7kg, time effect p[lt]0.001).[br]In both dietary groups, total urinary GC metabolite excretion tended to decrease (25%) over 24 months (time effect, p=0.07), with metabolite ratios indicating increases in both whole body 11[beta]HSD1 (24%) and 5[alpha]-reductase (40%, time effect, p[lt]0.001) activities. SAT 11[szlig]HSD1 mRNA decreased during the first 6 months (20%) in the PD group (diet x time effect, p[lt]0.05), and between 6 and 24 months (17%) in the NNR group (time effect, p[lt]0.05), but otherwise there were no differences between diets. No association between changes in GC metabolism and anthropometry or glucose/insulin levels were found.[br]Long-term weight loss induced by dietary intervention in postmenopausal women reverses 11[beta]HSD1 alterations seen with obesity, but increases 5[alpha]-reductase activity. These effects are independent of differences in macronutrient proportions in the diet and of altered plasma insulin, and not associated with alterations in anthropometry (e.g BMI, fat mass), suggesting they are mediated by other factors (such as intra-adipose inflammation).[br][br]Nothing to Disclose: AP, KS, MR, RA, BW, TO 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 798 179 1194 SUN-137 PO28-03 Sunday 989 2012


988 ENDO12L_SUN-138 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Effect of Omentectomy Associated to Roux-en-Y Gastric Bypass on Whole-Body Insulin Sensitivity and Beta-Cell Function Marcelo Miranda de Oliveira Lima, Jose Carlos Pareja, Sarah Monte Alegre, Sylka Rodovalho Geloneze, Steven E Kahn, Brenno Astiarraga, Elinton Adami Chaim, Bruno Geloneze State University of Campinas (UNICAMP), Campinas, Brazil; Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, Brazil; Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, Brazil; VA Puget Sound Health Care System and University of Washington, Seattle, WA The visceral fat is linked to insulin resistance, the metabolic syndrome, type 2 diabetes and an increased cardiovascular risk, but it is not clear whether it has a causative role. Surgical resection of this fat depot is a research model to address this issue. To describe the additive effect of omentectomy over Roux-en-Y Gastric Bypass (RYGBP) on insulin sensitivity and beta-cell function, 20 premenopausal women (35.29[plusmn]6.74 year-old, BMI, 45.51[plusmn]3.74 kg/m2) with metabolic syndrome were randomized to either RYGBP alone (CT, n=10) or combined with total omentectomy (OM, n=10). Normal glucose tolerance, impaired glucose tolerance and type 2 diabetes (HbA1c[lt]7% without glucose-lowering agents) were found in CT (respectively, n=4/3/3) and OM (n=3/4/3). Insulin sensitivity (IS; euglycemic-hyperinsulinemic clamp), acute insulin response to glucose (AIR; intravenous glucose tolerance test), disposition index (DI = AIR x IS), lipid profile, adipokine profile (leptin, adiponectin, resistin, visfatin, interleukin-6, TNF-[alpha], MCP-1), ultra-sensitive C-reactive protein (CRP), body composition and abdominal fat echography were assessed at baseline and 1, 6-8 and 12-18 months postoperatively. OM had greater weight loss at all time points. IS improved similarly in both groups. OM had lower CRP after 12 months [0.05 (0.05) vs 0.26 (0.23) mg/dL], but there was no difference in adipokines and other metabolic parameters. Among non-diabetic subjects, omentectomy was associated with a preservation of baseline AIR after 12 months [from 3.09 (0.79) at baseline to 2.91 (0.61) after 12 months] as opposed to a deterioration in the control group [from 3.19 (1.52) to 1.51 (0.67) pmol/l x10min x 103]; and a greater DI after 6 [141.15 (40.85) vs 75.88 (35.38)] and 12 months [166.61 (38.07) vs 88.8 (46.88) pmol/l x 10 min x [mu]mol/kg[sub]FFM[/sub]/min x 10[sup]3[/sup])]. Although it did not enhance the effect of RYGBP on insulin sensitivity and adipokines, omentectomy was associated with a preservation of insulin secretion, a greater weight loss and lower CRP. An influence of visceral fat on weight control and beta-cell function through endocrine/inflammatory pathways is hypothesized.[br][br]Sources of Research Support: The present study was funded by Funda[ccedil][atilde]o de Apoio [agrave] Pesquisa do Estado de S[atilde]o Paulo (FAPESP), S[atilde]o Paulo, Brazil (Protocol 05/58627-2) and supported in part by funding from the United States Department of Veterans Affairs.[br][br]Nothing to Disclose: MML, JCP, SMA, SRG, SEK, BA, EAC, BG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1599 179 1195 SUN-138 PO28-03 Sunday 990 2012


989 ENDO12L_SUN-139 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Changes in Gastrointestinal Transit in the Setting of Caloric Restriction with or without Adjustable Gastric Banding Matheni Sathananthan, Jennie Law, Paula Giesler, Jeanette Laugen, Michael Camilleri, Duane Burton, Adrian Vella Mayo Clinic, Rochester, MN; Mayo Clinic, Rochester, MN Prior studies have shown that gastric emptying is slowed after several days of fasting but it is unclear the effects of caloric restriction on gastric emptying over time. The effects of adjustable gastric banding (AGB) on gastric emptying are also uncertain. Given that caloric restriction may be the main mechanism by which AGB mediates its effects it is important to characterize changes in gastrointestinal function due to caloric restriction alone or in combination with AGB. We hypothesized that in people with type 2 diabetes, caloric restriction delays gastric emptying. We studied subjects with type 2 diabetes who underwent AGB (n=5) and matched diabetic controls (n=8) at baseline, 6 [amp] 12 weeks after initiation of caloric restriction similar to that which patients follow after AGB. All subjects consumed a pureed diet ([sim]800Kcal/day) over the study period. During each study day, subjects consumed a mixed meal consisting of one scrambled egg, 15g of ham and 35g of Jell-O labeled with [1-[sup]13[/sup]C]-glucose. To enable measurement of solid phase gastric emptying and orocecal transit, the egg was labeled with 0.1 mCi [sup]111[/sup]InCl[sub]3[/sub] activated charcoal. Gastrointestinal and colonic transit was measured by anterior and posterior gamma camera images obtained immediately after meal ingestion, every 15 minutes for the first 2 hours, then every 30 minutes for the next 2 hours. The gastric emptying lag time (t[sub]lag[/sub]=time to empty 10% of gastric contents) and gastric emptying half-time (t[sub]50[/sub]=time to empty 50% of gastric contents) was calculated. The t[sub]lag[/sub] in the AGB group was 35.5[plusmn]8.3 vs. 43.4[plusmn]15.5 vs. 30.6[plusmn]16.2 minutes (baseline vs. 6 weeks vs. 12 weeks respectively, [italic]p[/italic]=0.85) and in the diet arm was 20.0[plusmn]8.8 vs. 22.38[plusmn]5.7 vs. 30.9[plusmn]10.5 minutes,[italic] p[/italic]=0.65. The t[sub]50[/sub] in the AGB arm was 116.8[plusmn]15.9 vs. 120.6[plusmn]21.9 vs. 101.8[plusmn]27.0 minutes, [italic]p[/italic]=0.82 and in the diet arm was 137.3[plusmn]16.7 vs. 146.0[plusmn]14.7 vs. 154.2[plusmn]12.6 minutes, [italic]p[/italic]=0.75. The baseline t[sub]lag[/sub] and t[sub]50[/sub] did not differ between the AGB and diet groups ([italic]p[/italic]=0.28 and [italic]p[/italic]=0.43 respectively). We conclude that in people with type 2 diabetes, caloric restriction alone or in combination with AGB does not alter gastric emptying.[br][br]Nothing to Disclose: MS, JL, PG, JL, MC, DB, AV 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 816 179 1196 SUN-139 PO28-03 Sunday 991 2012


990 ENDO12L_SUN-140 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Roux-en-Y Gastric Bypass in a Diet-Induced Rodent Model of Obesity and Its Effect on Glucose Homeostasis Raymond G Lau, Collin EM Brathwaite, Louis Ragolia Winthrop University Hospital, Mineola, NY; Winthrop University Hospital, Mineola, NY [bold][italic][underline]Background[/underline][/italic][/bold]: The remission of [italic]type 2[/italic] diabetes mellitus has been observed in upwards of 80% of those who undergo gastric bypass surgery (Buchwald H, [italic]et al.[/italic] 2009). Several mechanisms have been proposed to mediate this metabolic improvement including weight loss, increased insulin sensitivity, and altered gut hormone levels such as glucagon-like peptide-1 (GLP-1). We investigated the effects of the Roux-en-Y gastric bypass (RYGB) on key metabolic organs involved in glucose homeostasis.[br][bold][italic][underline]Methods[/underline][/italic][/bold]: A total of four Sprague Dawley rodents made obese on a high fat diet underwent RYGB, compared to four diet induced obese rodents that underwent sham surgery. Tissue biopsies were obtained at the time of surgery and at 10 weeks post-operatively. Fasting glucose, insulin, GLP-1, and glucose-dependent insulinotropic polypeptide (GIP) were measured just prior to surgery, 4 weeks, and 10 weeks post-surgery. RT-PCR methods were used to determine expression of mRNA levels of glucose-6-phosphatase (G6Pase), and phosphoenolpyruvate carboxylase (PEPCK) in hepatic tissue.[br][bold][italic][underline]RESULTS:[/underline][/italic][/bold] RYGB rodents lost as much as 25% of their pre-operative body weight after surgery. Fasting glucose in RYGB rodents was lower at 108 +/- 22.5 mg/dL as compared to sham surgery rodents 152 +/- 9.3 mg/dL at 10 weeks. Also in RYGB rodents, fasting insulin levels decreased from 2.02 +/- 0.22 ng/mL preoperatively to 0.88 +/- 0.13 mg/dL. HOMA-IR (Homeostasis Model Assessment of Insulin Resistance) also decreased after RYGB from 5.8 to 2.9. Fasting total GLP-1 levels increased from 18.5 +/- 2.74 pM prior to surgery, to 4 weeks 53.8 +/- 5.08 pM, and 10 weeks 81.2 [bold]+/- [/bold]40.4 pM. However, evaluation of gluconeogenic enzymes revealed the mRNA expression of hepatic G6Pase was 1.6-fold elevated after surgery, as compared to prior to surgery. PEPCK mRNA expression also showed no difference before and after surgery.[br][bold][italic][underline]CONCLUSION[/underline][/italic][/bold]: Few have studied the temporal changes in fasting gut hormone levels after gastric bypass in rodent models and their effects on liver, pancreas, and skeletal muscle. Although not observed in humans, elevation in fasting gut hormone levels have been observed after RYGB as compared to sham surgery rodents (Shin AC et al 2011). The potential improvement of glucose metabolism following RYGB is possibly mediated by increasing fasting GLP-1 levels. Hepatogluconeogenesis does not appear to have a direct role, although the liver has at least a partial role as there is improved hepatic insulin sensitivity.[br][br]Buchwald H, Estok R, Fahrbach K, Banel D, Jensen MD, Pories WJ, Bantle JP, Sledge I. Weight and type 2 diabetes after bariatric surgery: systematic review and meta-analysis. Am J Med. 2009 Mar;122(3);248-256. Shin AC, Zheng R, RL Townsend, David L Sigalet, and HR Berthoud. Meal Induced Hormone Responses in a Rat model of Roux-en-Y Gastric bypass surgery. Endocrinology, April 2010, 151(4): 1588-1597.[br][br]Nothing to Disclose: RGL, CEMB, LR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1002 179 1197 SUN-140 PO28-03 Sunday 992 2012


991 ENDO12L_SUN-141 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Effect of High-Fat Diet[ndash]Induced Paternal Obesity on Physical Activity and Food Habits of Offspring Yuriy Slyvka, Yizhu Zhang, Alexis Zontini, Manindra Singh, Josh Ozbolt, Felicia V Nowak Ohio University, Athens, OH Epidemiological studies have shown that parental high fat diet (HFD) and increased body mass index correlate with abnormal metabolic profile and increased risk of obesity and insulin resistance in offspring. Based on this we studied physical activity and food habits in murine offspring that were born from obese male parents.[br]To develop obesity, C57BL/6 male mice were fed with HFD (45% kcal fat) for 12 weeks starting at the age of 4 weeks. After that they were mated with females on low fat diet (LFD, 10% kcal fat). Control group was formed of 8 mouse pairs with both parents on LFD. After weaning, all offspring were maintained on normal lab chow. Physical activity and stamina of offspring were tested using voluntary running wheel and involuntary swimming tests. Food preference (HFD vs. LFD) was determined and food and water consumption were measured.[br]There were no significant differences in voluntary (running wheel test) and involuntary (swim test) physical activity in offspring from male parents on HFD and control groups. Food and water consumption were equal in both groups of offspring. Food preference test results showed male offspring from group where male parents were on HFD consume more HFD than females from the same group or males from lean male parents. Although all groups showed a distinct preference for HFD, female offspring from the control group consumed only HFD. Other offspring groups consumed a small amount of LFD.[br]Conclusion: Offspring from obese and lean male parents displayed similar results in running wheel and swim test. Male offspring from obese male parents consumed more HFD in food preference test.[br][br]Nothing to Disclose: YS, YZ, AZ, MS, JO, FVN 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 986 179 1198 SUN-141 PO28-03 Sunday 993 2012


992 ENDO12L_SUN-142 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Diagnostic Power of Baseline Leptin/Ghrelin Ratio in Predicting Weight Regain after Hypocaloric Diet[ndash]Induced Weight Loss Ana B Crujeiras, Angel Diaz-Lagares, Estibaliz Goyenechea, Itziar Abete, Maria Amil, J Alfredo Martinez, Felipe F Casanueva Instituto de Investigaci[oacute]n Sanitaria, Complejo Hospitalario Universitario de Santiago (CHUS), Santiago de Compostela, Spain; Instituto de Salud Carlos III (ISCIII), Madrid, Spain; Complejo Hospitalario Universitario de Santiago (CHUS), Santiago de Compostela, Spain; University of Navarra, Pamplona, Spain Context: Weight regain after hypocaloric diet-induced weight loss has emerged as one of the most significant obstacles for obesity therapy (1). Therefore, the identification of factors associated with weight regain would have positive implications in the personalization of therapeutic approaches of obesity. Previous results from our research group suggest the existence of two predetermined different populations of patients according to the baseline leptin and ghrelin levels (2). For these reasons, the leptin/ghrelin ratio could be proposed as a marker of future success in the nutritional treatment outcome.[br]Objective: The aim of this study was to evaluate the capacity of the leptin/ghrelin ratio as a non-invasive tool for early identification of those obese patients more prone to regain the lost weight after an energy restriction program and to assess the leptin/ghrelin ratio cut-off that may help to differentiate weight loss regainers and non-regainers between obese patients.[br]Methods[bold]:[/bold] Baseline fasting leptin and ghrelin levels were evaluated and the L/G ratio was calculated in a group of overweight/obese patients (40 regainers and 48 non-regainers) that followed a successful 8-wk hypocaloric diet. The receiver operating characteristic (ROC) curve analysis was employed to evaluate the diagnostic value of the L/G ratio and to establish a cut-off point to differentiate regainers and non-regainers of [ge]10% weight loss 32 weeks after finishing the dietary treatment.[br]Results: Men and women weight regainers showed statistically two-fold higher leptin/ghrelin ratio at baseline (L/G[sub]B[/sub]) than non-regainers. L/G[sub]B[/sub] ratio directly correlated with weight loss regain (r=0.30; p=0.005). Analyses of ROC curves demonstrated that weight-loss regain can be predicted with 70% sensitivity in women and 95% in men, as well as with 68% specificity in women and 38% in men with the use of the optimal identified L/G[sub]B[/sub] ratio cut-off of 26.0 for women and 9.5 for men (women area under the ROC curve (AUC)=0.69; P=0.040 and men AUC=0.68; P=0.030).[br]Conclusions: Measurement of circulating fasting leptin/ghrelin ratio may offer a noninvasive approach that could help health professionals to avoid unsuccessful treatment outcomes and to personalize obesity therapy.[br][br](1) Sumithran P et al., N Engl J Med. 2011;365:1597. (2) Crujeiras AB et al., J Clin Endocrinol Metab. 2010;95:5037.[br][br]Sources of Research Support: Research relating to this abstract was funded by the Linea Especial about Nutrition, Obesity and Health (University of Navarra LE/97); CIBERobn and RETICS/PREDIMED, Instituto de Salud Carlos III (ISCIII) iniciatives. AB Crujeiras and I Abete are funded by the Instituto de Salud Carlos III (ISCIII) through a research-staff contract [ldquo]Sara Borrell[rdquo]. A Diaz-Lagares is funded by the Xunta de Galicia through a research-staff contract [ldquo]Angeles Alvari[ntilde]o[rdquo].[br][br]Nothing to Disclose: ABC, AD-L, EG, IA, MA, JAM, FFC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 300 179 1199 SUN-142 PO28-03 Sunday 994 2012


993 ENDO12L_SUN-143 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Dietary Fat Oxidation in Lean, Obese and Reduced-Obese Male and Female Adults Audrey Bergouignan, Elizabeth H Kealey, Stacy L Schmidt, Matthew R Jackman, Daniel H Bessesen University of Colorado, School of Medicine, Denver, CO Although a defect in fat oxidation due to altered oxidative capacity has been suggested by many to be a central mechanism leading to obesity, recent studies have challenged this concept. This controversy likely relates in part to the subjects studied and methods used to measure fat oxidation. While many previous studies have used indirect calorimetry to assess total fat oxidation, tracer methods may provide more direct information about the nature of the defect if present. We tested the hypothesis that both total and dietary fat oxidation is decreased in obese and moreso in reduced-obese compared to lean individuals. We measured 24hr total (room calorimeter) and dietary fat oxidation (following the ingestion of a test meal containing [sup]14[/sup]C-oleate) in sedentary lean (n=10), obese (n=9) and reduced-obese ([gt]10% of initial body weight; n=7) adult men and women. Reduced-obese were thinner than obese individuals (mean[plusmn]sd; BMI=26.9[plusmn]3.7 vs 33.6[plusmn]2.5 kg/m[sup]2[/sup] and %BF=30.0[plusmn]6.1% vs 38.1[plusmn]7.3%, p[lt]0.05) but fatter than lean individuals (BMI=26.9[plusmn]3.7 vs 21.5[plusmn]1.6 kg/m[sup]2[/sup] and %BF=30.0[plusmn]6.1% vs 22.5[plusmn]6.2%, p[lt]0.05). Although 24hr total fat oxidation was 28% lower in reduced-obese individuals compared to both lean and obese groups (p=0.06 and p=0.04, respectively), the amount of meal fat oxidized over 24hr did not differ between groups (6.0[plusmn]1.3g, 6.7[plusmn]1.8g, 6.1[plusmn]2.4g in lean, obese and reduced-obese respectively), even after adjustment for fat-free mass (FFM) and energy balance. While no correlation was observed between FFM and total fat oxidation, FFM accounted for 35% of the variation in dietary fat oxidation among the groups (p[lt]0.0001), suggesting that total and dietary fat oxidation are regulated differently. In conclusion, we found no difference in fat oxidation between sedentary obese individuals and sedentary lean individuals. These results do not support the hypothesis that muscle oxidative capacity is reduced in obese individuals. However, fat oxidation was lower in reduced-obese individuals which could contribute to weight regain. This defect was not related to a decrease in dietary fat oxidation suggesting a decrease in plasma free fatty acid or intra-myocellular fat oxidation. Further studies are needed to determine the respective roles of adipose tissue and muscle in the altered fat metabolism seen following weight loss.[br][br]Sources of Research Support: NIDDK K24 DK02935.[br][br]Nothing to Disclose: AB, EHK, SLS, MRJ, DHB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2145 179 1200 SUN-143 PO28-03 Sunday 995 2012


994 ENDO12L_SUN-144 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Predictors of Weight Gain after Orthotopic Liver Transplant at 2 Years Jasmine Kouz, Agnes Rakel, Catherine Vincent McGill University, Montr[eacute]al, Canada; University of Montr[eacute]al, Montr[eacute]al, Canada Introduction: Weight gain post transplant is well established in orthotopic liver transplant (OLT). Obesity increases cardiovascular morbidity and mortality in this population, but might also affect the new liver. We aimed to determine whether patient with NASH cirrhosis had a higher weight gain after OLT compared to other type of cirrhosis and to evaluate markers predicting patients at greater risk of weight gain.[br]Methods: We conducted a retrospective study of 126 liver transplant recipients from January 2005 to December 2007 who had a two years follow up at Saint-Luc[apos]s Hospital in Montreal. Patients[apos] weight were evaluated at baseline, 3, 6, 12 and 24 months after transplant. We collected data on age at transplant, sex, ethnic group, smoking status, the presence of hepatoma or ascites before OLT, the type of cirrhosis, the length of hospitalization after OLT, the presence of diabetes before and during follow up, the level of cholesterol and triglycerides before and during follow up, the cumulated dose of corticosteroids during follow up, the type of immunosuppressive medication used at two years and the incidence of different complications after OLT such as reject.[br]Results: We studied 17 patients who had a NASH cirrhosis compared to 109 patients with other types of cirrhosis. Mean age was 52 years, with 57% of men and 86% of Caucasian. There was no significant difference in weight gain between NASH and non NASH patients at one or two years when compared to baseline weight, but compared with the third month post OLT weight, which is free of ascitis, weight gain was significatively more in the NASH patients. Both in the univariate and the multivariate linear regression analysis adjusted for age, sexe, NASH cirrhosis, BMI before OLT, the presence of ascites, the cumulated dose of corticosteroids, weight gain at one and two years was significantly greater in the NASH group(p=0,001 and p=0,049 respectively) and patient who had ascitis(p=0,004 and p=0,002).[br]Conclusion: NASH cirrhosis patients had a greater weight gain two years post transplant, which was also more rapid in the first year. This should emphasize the need of a tight follow up along with nutritionist supervision and exercise program in post transplant patients, particularly NASH patients.[br][br](1) Paul Angulo, Liver Transplantation, 2006; 12:523. (2)Suk Seo et al, Liver Transplantation, 2007; 13:844. (3)Wawrzynowicz-Syczewska M et ak, Ann Transplant. 2009;14:45. (4)Yalamanchili K et al, Liver Transpl. 2010;16:431. (5) Contos MJ et al, Liver Transplant 2001;7:363.[br][br]Nothing to Disclose: JK, AR, CV 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 991 179 1201 SUN-144 PO28-03 Sunday 996 2012


995 ENDO12L_SUN-145 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Central and Peripheral Endocannabinoid Levels in Obese Versus Lean Women Mariana Farage, Rosilane Taveira da Silva, Denise Pires Carvalho, Amelio Fernando Godoy-Matos Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil; Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil; Instituto de Diabetes e Endocrinologia Luiz Capriglione, Rio de Janeiro, Brazil Introduction: Mechanisms involving body weight regulation and appetite control have being extensively studied in the last years. The endocannabinoid system (ECS) seems to play an important hole in obesity and its activation increases appetite and weight gain (1,2). The ECS is expressed both in central nerve system (CNS) and peripheral tissues, mainly visceral adipocytes, muscle and pancreas (3). ECS is hyperactivated in the hypothalamus of obese mice (1). Peripheral overactivation of ECS has been demonstrated in humans (4,5) and circulating 2-AG correlated with body fat, visceral fat and fasting glucose concentration (6). There is no data about ECS activity in human CNS.[br]Aims: To test the hypothesis that ECS is hyperactivated in human CNS.[br]Methods: Thirteen obese and 11 lean women submitted to elective surgery under raquianesthesia were evaluated. Cerebral spinal fluid (CSF) and blood samples were collected for 2-arachidonoyl glycerol(2-AG) and anandamide (AEA) levels analyses.[br]Results: AEA was decreased in CSF of obese women (25,44 [plusmn] 8,33 vs 49,89 [plusmn] 3,38 fmoles/mL p[lt]0,05) while it was increased in plasma(4,418 [plusmn] 0,9792 pmoles/mL vs 1,890 [plusmn] 0,197 pmoles/mL p[lt]0,05). Plasma/CSF ratio was 41.58[plusmn]5.78 (n=10) for lean and 103.0[plusmn]37.36 (N=6) for obese (p=0,0543). There was no correlation between plasma-AEA and CSF-AEA. Analysis of 2-AG was not possible.[br]Discussion: Our data suggest that the activation of ECS in obesity is more peripheral than central. Indeed, it seems to be suppressed in the CNS of obese women. Peripheral activity of ECS may be more relevant for obesity than CNS. Ravinet Trillou et al. have shown that fat mass loss under chronic CB1R blockade was unrelated to energy intake in a mouse model of diet-induced obesity (7). Treatment of mice with AM6545, a neutral CB1R antagonist selective to peripheral tissues, lead to weight reduction without affecting caloric intake (8). This data support the hypothesis that ECS activation and deregulation in peripheral tissues may be key in obesity. Therefore, targeting peripheral ECS, and so avoiding CNS adverse effects, seems to be promising.[br]Acknowledgements:We thank Dr. George Kunos for measuring anandamide levels by LC/MS/MS.[br][br](1) Di Marzo V et al (2001) Leptin-regulated endocannabinoid are involved in maintaining food intake. Nature 410:822-825. (2) Di Marzo V, Matias I (2005) Endocannabinoid control of food intake and energy balance. Nat Neurosci 8(5):585-9. (3) Pacher P, Batkai S, Kunos G (2006) The endocannabinoid system as an emerging target of pharmacotherapy. Pharmacol Rev 58:389-462. (4) Matias I et al (2006) Regulation, Function, and Dysregulation of Endocannabinoids in Models of Adipose and Pancreatic Cells and in Obesity and Hyperglycemia. J Clin Endocrinol Metab 91:3171[ndash]3180. (5) Cote M et al (2007). 2-Arachidonoylglycerol in abdominally obese males. International Journal of Obesity 1[ndash]8. (6) Bluher M et al (2006). Dysregulation of the Peripheral and Adipose Tissue Endocannabinoid System in Human Abdominal Obesity. Diabetes 55: 3053-3060. (7) Ravinet Trillou C, Arnone M, Delgorge C, Gonalons N, Keane P, Maffrand JP, Soubrie P (2003). Anti-obesity effect of SR141716, a CB1 receptor antagonist, in diet-induced obese mice. Am J Physiol Regul Integr Comp Physiol 284: R345[ndash]R353. (8) Kunos G, Tam J (2011) The case for peripheral CB 1 receptor blockade in the treatment of visceral obesity and its cardiometabolic complications. British Journal of Pharmacology 163: 1423[ndash]1431.[br][br]Nothing to Disclose: MF, RTdS, DPC, AFG-M 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1158 179 1202 SUN-145 PO28-03 Sunday 997 2012


996 ENDO12L_SUN-146 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Resveratrol Prevents Hyperleptinemia and Central Leptin Resistance in Adult Rats Programmed by Early Weaning Elaine de Oliveira, Patricia C Lisboa, Juliana G Franco, Isis H Trevenzoli, Natalia L da Silva, Nayara Peixoto-Silva, Egberto G Moura State University of Rio de Janeiro, Rio de Janeiro, Brazil; Federal University of Rio de Janeiro, Rio de Janeiro, Brazil [bold]Background:[/bold] Obesity and its clustering of cardiovascular risk factors is increasing worldwide. Exclusive breastfeeding during six months has been associated with obesity prevention. Resveratrol has been shown to improve cardiovascular disease and much speculation about its use in the treatment of obesity has emerged in the last years. [bold]Objectives: [/bold]In this study, we evaluated the effects of resveratrol (RES) over visceral obesity, hyperleptinemia and insulin and leptin resistance in a developmental plasticity model of obesity in adult early weaned rats. [bold]Methods: [/bold]To induce early weaning, lactating dams were separated into two groups: early weaning (EW) [ndash] dams were wrapped with a bandage to interrupt lactation in the last 3 d of lactation and control (C) [ndash] dams whose pups had free access to milk during the entire lactation period (21 d). At 150 days-old, EW animals were subdivided into: EW+RES (30mg/kg/BW) during 30 days and EW treated with diluent solution both by gavage. Significant differences had p[lt]0.05. [bold]Results:[/bold] EW rats showed higher body weight (9%), hyperphagia (18%), visceral obesity (44%), hyperleptinemia (64%), hyperglycemia (38%), insulin resistance and hypoadiponectinemia (22%) at adulthood that was prevented with resveratrol treatment. Leptin resistance associated with lower JAK2 (27%) and pSTAT3 (40%) and higher NPY (39%) in hypothalamus of EW rats was normalized by resveratrol, and these seem to be independent of SOCS3 normalization. [bold]Conclusions: [/bold]The present results suggest that resveratrol is useful as therapeutic tool in treating obesity as this compound could modulate fat accumulation and appetite control.[br][br]Sources of Research Support: FAPERJ, CNPQ and CAPES.[br][br]Nothing to Disclose: EdO, PCL, JGF, IHT, NLdS, NP-S, EGM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1223 179 1203 SUN-146 PO28-03 Sunday 998 2012


997 ENDO12L_SUN-147 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) IGF2 Gene Polymorphisms That Alter Circulating IGF-II Levels May Determine Weight Loss in Diabetes Ram Prakash Narayanan, Simon G Anderson, Bo Fu, Anthony Payton, Julie E Hudson, Robert L Oliver, Kirk W Siddals, John P New, Anne White, Adrian H Heald, William ER Ollier, John Martin Gibson The University of Manchester, Salford, UK; The University of Manchester, Manchester, UK; The University of Manchester, Manchester, UK; The University of Manchester, Manchester, UK; The University of Manchester, Manchester, UK; The University of Manchester, Manchester, UK Low IGF-II has an established association with future weight gain (1), and the IGF2 gene has been linked with obesity(2). We aim to identify whether variants in IGF2 could predict changes in body mass index (BMI) as well as circulating IGF-II.[br]We studied fifteen haplotype tagging single nucleotide polymorphisms (SNPs) from the IGF2 gene in 487 Caucasian individuals with type 2 diabetes. ht-SNP selection used HapMap, Haploview, Ensembl and Tagger. Quality assurance was done on SVS7 (call rate[gt] 90%, minor allele frequency [ge]10% and HW equilibrium). BMI data was obtained for the period 2002 to 2009 from primary and secondary care data. Serum IGF-II was measured with a local extraction based ELISA on blood samples obtained once in 2002-03. Age and gender adjusted regression analyses of SNPs with IGF-II levels was done using an additive model in STATA 10SE. A mixed-effects regression model was used to study longitudinal trends of SNPs with BMI adjusted for age and gender.[br]The minor allele of the rs3842767 IGF2 SNP predicted a decrease in serum IGF-II (for log IGF-II: [beta] = -0.084, [95% CI -0.15 to -0.013], p=0.02) independent of age and gender. Addition of each minor allele influenced an increase in BMI of 0.11 kg/m2 in an average year compared to the homozygous major allele ([beta] 0.11 [95% CI 0.01 to 0.21] p=0.025).[br]The variant allele of the rs10770063 SNP intergenic to IGF2 and H19 was associated with increased serum IGF-II levels independent of age and gender (for log IGF-II: [beta]=0.049, [95% CI: 0.006 to 0.091], p=0.023). Each minor allele was associated with a decrease in BMI adjusted for age and gender ([beta] -0.07 [95% CI: -0.12 to -0.01] p=0.014).[br]We report novel associations of polymorphisms of IGF2 with concentration of circulating IGF-II and also with longitudinal weight change. This indicates that IGF2 polymorphisms may be important in determining longitudinal weight trends in type 2 diabetes.[br][br](1) Heald AH et al.,Am J Med 2006 Feb; 119(2): 167. (2) Gaunt TR et al.,Hum Mol Genet. 2001 Jul 1;10(14):1491-501.[br][br]Nothing to Disclose: RPN, SGA, BF, AP, JEH, RLO, KWS, JPN, AW, AHH, WERO, JMG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1419 179 1204 SUN-147 PO28-03 Sunday 999 2012


998 ENDO12L_SUN-148 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Sexual Dimorphic Response to Maternal High-Fat Diet Katelyn Elizabeth Finch, Reina Mayor, Lisa Hahner, Deborah Clegg University of Texas Southwestern Medical School, Dallas, TX There is an emerging childhood obesity epidemic with over 43 million children world-wide under the age of five which are now considered to be overweight by the World Health Organization. Increased availability and consumption of high-fat foods is a major component to the global epidemic of increased body weight and body fat stores, contributing to the development of diseases such as obesity, diabetes, cardiovascular disease, and cancers. Additionally, it has recently been appreciated that males and females differ with respect to disease prevalence and metabolism. What is not known is how the maternal diet influences the programming of the fetal metabolism, which is setting the stage for children to become obese. Therefore, our goal has been to determine if there is a sexually dimorphic response to the impact of the maternal diet on fetal programming of metabolic diseases. We analyzed placental gene expression from female and male fetuses taken from C57BL6 mice exposed to a high fat diet. We focused our attention on inflammatory pathways, since it is known that saturated fatty acids activate pathways such as the toll-like receptor 4 (TLR4) pathway. We found in female fetal placentas that the TLR4 pathway was significantly activated when compared to the males. From these data, we hypothesized that there was a sexual dimorphism with respect to the transport and storage of fatty acids (FA); therefore, we evaluated body adiposity in female and male fetuses by X-ray computed tomography. We found increased total body fat in females relative to males, and this was independent of differences in body weight. It is well established that insulin and leptin secretion changes in direct proportion to adiposity; therefore, we analyzed the concentration of insulin and leptin present in the amniotic fluid from both female and male fetuses by ELISA, and again we saw an increased concentration of insulin and leptin only in the females. This suggests that maternal HFD differentially affects males and females. There is increased transport and storage of FA in the females, whereas the males are relatively protected from the FA and have reductions in body adiposity. Both an increase and decrease in adiposity in utero can predispose the offspring to metabolic disease, and since the placenta is a direct mediator of nutrient supplies, future research should focus on understanding the effect of HFD on placental function and fetal programming associated with metabolic diseases.[br][br]Nothing to Disclose: KEF, RM, LH, DC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1964 179 1205 SUN-148 PO28-03 Sunday 1000 2012


999 ENDO12L_SUN-149 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Additive Effects of Nicotine and High-Fat Diet on Hepatic Steatosis in Mice Are Mediated by Abdominal Lipolysis and Involve Oxidative Stress and Hepatocellular Apoptosis Amiya Sinha-Hikim, Indrani Sinha-Hikim, Meher Parveen, Michael Mangubat, Chang-Sung Shin, Alexei Lyzlov, Rasheed Ivey, Samuel French, Theodore Friedman Charles R Drew University of Medicine [amp] Science, Los Angeles, CA; LA-Biomed, Torrance, CA Smoking is a major risk factor for diabetes and cardiovascular disease and may contribute to non-alcoholic fatty liver disease (NAFLD). NAFLD has been identified as an independent risk factor for insulin resistance, atherosclerosis and cardiovascular disease. We hypothesized that nicotine plus a high-fat diet (HFD) would have additive effects on the severity of hepatic steatosis in obese mice. Adult C57BL6 male mice were fed with normal chow diet (NCD) or HFD with 60% of calories derived from fat with twice daily injections of nicotine (0.75 mg/kg BW, ip) or saline for 10 weeks. Light and electron microscopy revealed markedly increased lipid accumulation in hepatocytes from mice on a HFD plus nicotine, compared to mice on a HFD alone. The additive effects of nicotine plus a HFD was associated with greater oxidative stress, increased incidence of hepatocyte apoptosis, inactivation (dephosphorylation) of AMPK, and activation of its downstream target, acetyl- COA-carboxylase (ACC), leading to increased lipogenesis. Treatment with acipimox (an abdominal lipolysis inhibitor) completely blocked nicotine plus HFD-induced hepatic fat accumulation. We conclude that: 1) greater oxidative stress coupled with inactivation of AMPK may be critical for the additive effects of nicotine and HFD on the severity of hepatic steatosis in obese mice and 2) lipolysis of abdominal fat is an important contributor to the accumulation of lipids in the liver. We surmise that nicotine plus HFD is likely to be a very toxic combination in patients.[br][br]Nothing to Disclose: AS-H, IS-H, MP, MM, C-SS, AL, RI, SF, TF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2073 179 1206 SUN-149 PO28-03 Sunday 1001 2012


1000 ENDO12L_SUN-150 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Feeding Behavior in Obese School Children after Low-Fat or Low-Carbohydrate Diet Ibarra Lorena, Pisarchik Lidmila, Juan M Malacara, Ma Eugenia Garay-Sevilla, Elva L Perez-Luque University of Guanajuato, Leon, Mexico [bold]Introduction:[/bold] Feeding behavior has important aspects to understand the development and maintenance of obesity. The Children[apos]s Eating Behavior Questionnaire (CEBQ) measures attitudes toward food restriction in obese children.[br][bold]Objective:[/bold] to evaluate feeding behavior in obese children under two types of diet restriction.[br][bold]Material and Methods:[/bold] We studied prospectively two groups of 60 school children (6 to 12 years of age) under a low carbohydrate (L-CHO) or low fat diet (L-F) during two months. We examined anthropometric and metabolic measurements, and the CEBQ scale, before and after intervention. Differences were analyzed with ANOVA for repeated measurements and the paired t test.[br][bold]Results:[/bold] After two months of dietary restriction BMI decreased 1.2 kg/m[sup]2[/sup] in both groups. Percent body fat, waist/hip ratio and HOMA-IR value diminished (p[lt]0.02, p[lt]0.008 and p[lt].03, respectively) and adiponectin increased (p[lt].006) only for the L-CHO diet. Glucose, cholesterol, and triglycerides did not change with treatment. After one month of treatment children showed decreased indices for satiety (14.2[plusmn]4.3 to 12.6[plusmn]4.4; p[lt] 0.00007), food fussiness (de 12[plusmn]3 to 10.6[plusmn]2.7; p[lt] 0.00004), responsiveness (11.4[plusmn]4.8 to 8.4[plusmn]4.9; p[lt].0000001) and enjoyment of foods (12.0[plusmn]2.6 to 10.5[plusmn]2.7; p[lt]0.0000001), this effect was maintained after two months in both groups. No difference in feeding behavior was found between both groups of intervention or by gender.[br][bold]Conclusions:[/bold] Our results show that under L-F or L-CHO diet, obesigenic behavior increases in school children. These findings may indicate that the set point for energy intake is not modified after two months of caloric restriction, and it is an important factor hindering weight loss.[br][br]Sources of Research Support: In part by Grant FOMIXGTO 10-16-A-036.[br][br]Nothing to Disclose: IL, PL, JMM, MEG-S, ELP-L 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1349 179 1207 SUN-150 PO28-03 Sunday 1002 2012


1001 ENDO12L_SUN-151 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Comparison of Serum Leptin and Adiponectin in Lean, Obese and Gestational Diabetic Pregnancies Jose Meza, Christopher Maguire, Urvi Shah, Blanca Martinez, Glena Davis, V Daniel Castracane Texas Technical Health Science Center, Odessa, TX [bold]Objective:[/bold] Obesity, either prepregnancy or weight gained during gestation are documented risks for the development of gestational diabetes (GDM). Pregnancy has long been described as an insulin resistant state and changes in adipokines may play a role in this condition over the course of gestation. GDM has been associated with increased leptin and decreased adiponectin levels and it has been suggested that these adipokines may have a diagnostic potential for GDM. In this study we have examined maternal serum levels of leptin and adiponectin in lean, obese and gestational diabetic pregnancies.[br][bold]Materials and Methods[/bold]: These results were generated from 3 different but similar protocols which obtained serum either in a longitudinal or cross sectional design in pregnant women that were lean, obese or GDM. Data will be presented for serial data from lean and obese women from five separate windows during gestation and in addition, also combined with cross sectional data for third trimester analysis which includes GDM subjects. Protocols were approved by the TTU IRB and subjects gave their written informed consent. Leptin and adiponectin were determined using specific ELISA assays (Millipore).[br][bold]Results[/bold]: Serial results show an increase in serum leptin with higher levels in the obese group and adiponectin levels decrease during gestation with lower levels in the obese group. In the larger group adding the cross sectional samples at the third trimester, mean leptin levels are low in the lean group (39.3 ng/ml: n=31) and significantly increased in the obese and GDM group (52.7 ng/ml; n=19 and 51.5 ng/ml; n=10, respectively) but not significantly different. Mean adiponectin levels are highest in the lean group (16.2 ug/ml) and significantly decreased in the obese and GDM group (10.03 ug/ml and 10.75 ug/ml, respectively) but not significantly different. [br][bold]Discussion[/bold]: During the course of pregnancy, obesity increases the levels of serum leptin and decreases the serum adiponectin levels. In these preliminary results, obese and GDM women in the third trimester have similar levels of leptin and adiponectin with no suggestion of a greater effect of GDM on these adipokine levels.[br][br]Nothing to Disclose: JM, CM, US, BM, GD, VDC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2010 179 1208 SUN-151 PO28-03 Sunday 1003 2012


1002 ENDO12L_SUN-152 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Leptin Suppression Is Impaired in Type 2 Diabetes Marlene Wewalka, Florencia Halperin, Raquel Manning, Rohit N Kulkarni, Allison B Goldfine Joslin Diabetes Center, Harvard Medical School, Boston, MA; Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA The effect of hyperinsulinemia on leptin production has remained controversial as multiple investigations yield conflicting results. Therefore we tested the effect of acute hyperinsulinemia on leptin concentration in three groups (11 subjects with T2D, 10 with IGT, and 9 healthy controls [Co]) by performing a 4-h isoglycemic-hyperinsulinemic clamp (2 mU insulin/kg/min) compared to a saline (sham) clamp as a time and volume control. Three leptin samples were drawn at 5 min intervals and averaged at baseline and at the end of the clamp procedure (240[sub]min[/sub]).[br]The three groups had different metabolic profiles with regard to baseline leptin (5.7[plusmn]1.6 [Co], 28.7[plusmn]7.8 [IGT], and 15.7[plusmn]5.6 ng/mL [T2D], [italic]P[/italic]=0.04), BMI ([italic]P[/italic]=0.05), fasting glucose ([italic]P[/italic]=0.0002), hemoglobin A1c ([italic]P[/italic][lt]0.0001), fasting insulin ([italic]P[/italic]=0.02), and clamp glucose utilization (M) (10.1[plusmn]0.7 [Co], 5.7[plusmn]0.9 [IGT], and 7.7[plusmn]0.4 [T2D], [italic]P[lt][/italic]0.001). IGT subjects were most obese, had highest fasting insulin and leptin, and were most insulin resistant, whereas T2D had highest fasting glucose, and hemoglobin A1c. During the isoglycemic-hyperinsulinemic clamp leptin increased in all groups with a similar magnitude of leptin increase in all groups (133[plusmn]8%, [italic]P[/italic][lt]0.02 [Co], 125[plusmn]11%, [italic]P[/italic]=0.075 [IGT] and 126[plusmn]6%, [italic]P[/italic]=0.03 [T2D]). During the sham clamp leptin levels decreased in all groups (-29[plusmn]3%, [italic]P[/italic][lt]0.01 [Co], [ndash]22[plusmn]6%, [italic]P[/italic]=0.02 [IGT], and -10[plusmn]4%, [italic]P[/italic][lt]0.01 [T2D]), however the magnitude of reduction in leptin between baseline and 240[sub]min[/sub] differed between the groups ([italic]P[/italic]=0.005) with a significantly blunted reduction in leptin concentration in T2D. The reduction in leptin between baseline and 240[sub]min[/sub] did not correlate with BMI, M, fasting glucose or insulin, or other metabolic or anthropometric characteristics. Leptin concentration increased during isoglycemic-hyperinsulinemic clamp and decreased during prolonged fasting (sham clamp) in all groups. The capacity to suppress leptin during fasting was different in T2D compared to healthy controls and IGT, while the change in leptin production during isoglycemic-hyperinsulinemic stimulation was similar in all groups. As the change in leptin during fasting was not correlated to any metabolic or anthropometric measure our findings suggest that the dynamics in leptin suppression was not caused by insulin resistance or fasting hyperglycemic stress but rather by other factors that are currently unknown.[br][br]Sources of Research Support: American Diabetes Association 06-CD-07 (ABG); National Institute of Health R01 DK070648 (ABG); R01 DK067536 (RNK); Marietta Blau Grant ICM-2010-02797 (MW); Joslin Diabetes and Endocrinology Research Center (DERC) P30-DK36836.[br][br]Nothing to Disclose: MW, FH, RM, RNK, ABG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1740 179 1209 SUN-152 PO28-03 Sunday 1004 2012


1003 ENDO12L_SUN-153 POSTER SESSION: Obesity Topics III (1:30 PM-3:30 PM) Leptin Induces Inflammatory Cytokine by PLC-[gamma][ndash]Dependent Metabolism of Arachidonic Acid Mediated by 5-LO and 12/15-LO in Alveolar Macrophages Semin Lee Hanyang University, Seoul, Republic of Korea Obesity is presumed to be associated with pathogenesis of asthma. Some kinds of asthma involve abnormal metabolism of arachidonic acid, resulting in an increase in 5-lipoxygenase (5-LO) metabolites. We investigated whether leptin, a classic adipocytokine, receptor take a role for increased 5-LO metabolites in mouse alveolar macrophage (Raw 264.7). Leptin treatment increased 5-LOand 12/15-LO expression in mouse alveolar macrophage. Leptin-induced 5-LO and 12/15-LO expression was attenuated by PLC-[gamma] inhibitor (PAO). Thus, we suggest leptin may contribute to development of asthma through increase metabolism of arachidonic acid mediated by 5-LO and 12/15-LO. Our result support obesity might contribute to pathogenesis of asthma.[br][br]Christine Sachs-Olsen et al, J ALLERGY CLIN IMMUNOL[apos] 2010,OCT, 859-867. Hyun-Jae Woo et al,Experimental Lung Research, 2010, 36, 262[ndash]269.[br][br]Nothing to Disclose: SL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2115 179 1210 SUN-153 PO28-03 Sunday 1005 2012


1004 ENDO12L_SUN-154 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) The Effects of Vitamin D and Calcium Supplementation on Cardiovascular Risk in Adults at High Risk for Diabetes Joanna Mitri, Bess Dawson-Hughes, Frank Hu, Anastassios G Pittas Tufts Medical Center, Boston, MA; Jean Mayer US Department of Agriculture Human Nutrition Research Center on Aging, Tufts University, Boston, MA; Harvard School of Public Health, Boston, MA and Channing Laboratory, Department of Medicine, Brigham and Women[apos]s Hospital and Harvard Medical School, Boston, MA Background: Observational studies suggest that vitamin D deficiency may modify risk of cardiovascular disease. However, the effect of vitamin D supplementation on cardiovascular risk factors in persons at risk for diabetes is unknown.[br]Objective: To determine whether vitamin D supplementation, with or without calcium, has an effect on lipid profile, endothelial markers, or blood pressure in adults at high risk for diabetes.[br]Design, setting and patients: Randomized placebo-controlled trial of ninety two adults with pre-diabetes randomized in a 2-by-2 factorial design to either cholecalciferol (2,000 international units once daily) or calcium carbonate (400 mg twice daily) for 16 weeks.[br]Outcomes: Change in total, LDL and HDL cholesterol, triglycerides, triglycerides to HDL ratio; endothelial markers (ICAM, VCAM and E-selectin); systolic and diastolic blood pressure over 16 weeks.[br]Results: Participants had a mean age of 57 years, BMI of 32 kg/m[sup]2[/sup] and hemoglobin A1c of 5.9%. There was no significant vitamin D x calcium interaction on any outcome. After adjustment for age, race, time of study entry, body fat percentage and physical activity, triglyceride concentration was lower in the vitamin D group compared to the no vitamin D group (adjusted mean change[plusmn]SEM -21.3[plusmn]9.1 mg/dl for vitamin D vs. 4.4[plusmn]8.5 for no vitamin D; p=0.021). The triglyceride-HDL ratio was also in the same direction (0.62[plusmn]0.27 mg/dl for vitamin D vs. 0.21[plusmn]0.26 for no vitamin D; p=0.014). There was no significant difference in the change in total, LDL or HDL cholesterol between vitamin D groups. E-selectin change was lower in the vitamin D group compared to the no vitamin D group (-2.9 [plusmn]0.8 ng/ml for vitamin D vs. -0.7[plusmn]0.8 for no vitamin D; p= 0.042) and there was no difference in change in ICAM or VCAM. There was no difference in change in blood pressure in any group. There were no differences in any of the outcomes between the calcium vs. no calcium groups.[br]Conclusion: In adults at risk for diabetes, short-term supplementation with cholecalciferol improved triglycerides, triglyceride-HDL ratio and E-selectin but had no effect on other vascular markers or blood pressure.[br][br]Sources of Research Support: NIH Research grant R01DK76092 (to AGP, funded by the National Institute of Diabetes and Digestive and Kidney Disease, and the NIH Office of Dietary Supplements); UL1 RR025752 (to Tufts Medical Center) from the National Center for Research Resources; the U.S. Department of Agriculture Cooperative Agreement No. 58-1950-4-401 (to BDH) and the Endocrine Fellows Foundation grant (to JM). Calcium carbonate pills and matching placebos were donated by Glaxo-Smith-Kline, Parsippany, NJ.[br][br]Nothing to Disclose: JM, BD-H, FH, AGP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2091 180 1211 SUN-154 PO10-01 Sunday 1006 2012


1005 ENDO12L_SUN-155 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Effects of Mixed Oral Glucose and Fat Load on Metabolic Parameters and Blood Pressure in Adolescents and Young Women with Polycystic Ovary Syndrome Hilda Carrilho Barbosa, Nara Crispim Carvalho, Paula Prazeres Aragao, Francisco Alfredo Bandeira Agamenon Magalh[atilde]es Hospital, University of Pernambuco Medical School, Recife, Brazil Polycystic ovary syndrome (PCOS) is usually associated with insulin resistance even in women who are not overweight. Post-prandial lipemia has been considered a marker of insulin resistance due to competition between chylomicrons and remnants particularly with endogenous (VLDL) triglycerides (TG). Few data are available regarding post-prandial lipemia in adolescents and young women with PCOS. [bold]Objective: [/bold]To assess the postprandial lipemic response after a mixed meal and its relationship to cardiovascular risk factors in adolescents and young adults, whether suffering or not from PCOS, by means of clinical, laboratory and inflammatory markers. [bold]Methodology: [/bold]We studied 40 young women (21 with PCOS and 19 non-PCOS). The mean age, body mass index, systolic blood pressure SBP, diastolic blood pressure DBP, TG, total cholesterol (TC), LDL-cholesterol, HDL-cholesterol and fasting glucose in the PCOS and non-PCOS groups were: 20.71 years (SD [plusmn]3.57) and 21.84 years (SD [plusmn]1.97), 23 Kg/m[sup]2 [/sup](SD [plusmn]5.19) and 19.5 Kg/m[sup]2 [/sup](SD [plusmn]2.47), 105.71 mmHg (SD [plusmn]16.9) and 101.58 mmHg (SD [plusmn]10.68), 69 mmHg (SD [plusmn]10.91) and 68,7 mmHg (SD [plusmn]9.4), 73.57 mg/dL (SD [plusmn]27.3) and 68.9 mg/dL (SD [plusmn]23,81), 154 mg/dL (SD [plusmn]20.51) and 158.9 mg/dL (SD [plusmn]21.7), 89.2 mg/dL (SD [plusmn]16.9) and 87.26 mg/dL (SD [plusmn]19.41), 50.53 mg/dL (SD [plusmn]13.02) and 58 mg/dL (SD [plusmn]12.24), 79.8 mg/dl (SD [plusmn]9.04) and 76.6 mg/dl (SD [plusmn]7.48), respectively. We measured TC and fractions, TG, insulin, glucose, leukocyte count and ultra-sensitive C-reactive protein after a 12-hour fast and 3 h and 5 h after the ingestion of a mixed meal containing 50 g of fat (milk cream: 25 g and butter: 24.6 g) and 50 g of carbohydrate (sugar: 22 g, guava jelly: 20 g and milk cream: 8.3 g). [bold]Results: [/bold]There were significant increases in TG (39% and 57%, p [lt] 0.001); serum insulin (80%, p=0.012 and 88%, p= 0.014) and in the leukocyte count (9%, p= 0.012 and 13%, p [lt] 0.001) in the PCOS and non-PCOS groups, respectively, when the data at three hours were compared. When triglyceridemia was correlated with SBP and DBP in PCOS and non-PCOS patients, a positive correlation was observed between the percentage variation in post-prandial triglyceridemia after three hours in relation to the baseline values with SBP (r=0.6295; p=0.022) and DBP (r=0.5539; p=0.00982) in the PCOS group. [bold]Conclusion: [/bold]We found a positive correlation between post-prandial lipemia responses after an oral fat load and blood pressure in nonobese, normolipemic and normotense young women with the PCOS.[br][br]Nothing to Disclose: HCB, NCC, PPA, FAB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1469 180 1212 SUN-155 PO10-01 Sunday 1007 2012


1006 ENDO12L_SUN-156 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Arterial Stiffness Is Associated with the Estimated GFR, but Not the Stage of Albuminuria, in Patients with Type 2 Diabetes Mellitus: A Cross-Sectional Study Kyung Yoon Chang, Ye Jee Lim, Won Chul Ha, Su-Jin Oh, Moo Il Kang, Hyun-Sihk Son, Tae Seo Sohn The Catholic University of Korea, Seoul, Korea Background: Microalbuminuria is well-established marker of increased CVD risk in patients with diabetes and cardiovascular event increases with the progression of the chronic kidney disease (CKD) stage based on estimated glomerular filtration rate (eGFR). The aim of this study is to assess the relationship between the arterial stiffness and carotid intima-media thickness (CIMT) and the stage of CKD and urinary albumin excretion in patients with type 2 diabetes mellitus (T2DM).[br]Methods: A cross-sectional study was performed in 490 patients with T2DM. The arterial stiffness was assessed by the brachial-ankle pulse wave velocities (baPWV) using a pulse wave velocimeter, CIMT was measured using an ultrasound examination, GFR was estimated by equation of simplified Modification of Diet in Renal Disease Study, and albuminuria was estimated by measurement of the albumin-to-creatinine ratio in a random spot collection.[br]Results: The subjects, whose age was 58 [plusmn] 11years, duration of diabetes 9 [plusmn] 8 years, HbA1c 9.13 [plusmn] 2.46 %, eGFR 77.91 [plusmn] 21.3 ml/min/1.73m[sup]2[/sup], and albuminuria 72 [plusmn] 188.5 mg/g. The baPWV was 1.73 [plusmn] 4.6 m/s, CIMT was 0.69 [plusmn] 0.26 mm. The baPWV correlated negatively with the eGFR (P[lt] 0.05) and positively with the CIMT (P[lt]0.05). The baPWV increased with the stage of CKD (1.65 [plusmn] 0.4 m/s in stage 1, 1.75 [plusmn] 0.49 m/s in stage 2, 1.78 [plusmn] 0.41 m/s in stage 3, and 1.85 [plusmn] 0.51 cm/s in stage 4+5). However, the PWV was not significantly different between the stage of albuminuria (1.72 [plusmn] 0.46 in normoalbuminuria, 1.70 [plusmn] 0.43 in microalbuminuria, 1.94 [plusmn] 0.64 in macroalbuminuria). The CIMT was not significantly different among the various stages of CKD and albuminuria.[br]Conclusions: Our study shows that arterial stiffness increases according to the stage of CKD, but not to the stage of albuminuria. The estimation of GFR in diabetic patients is essential not only in estimating renal function but also in estimating CVD risk.[br][br]Nothing to Disclose: KYC, YL, WCH, S-JO, MIK, H-SS, TSS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1437 180 1213 SUN-156 PO10-01 Sunday 1008 2012


1007 ENDO12L_SUN-157 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Development of a Sex- and Ethnicity-Specific Childhood Continuous Metabolic Syndrome Risk Score Matthew J Gurka, Christa L Ice, Shumei S Sun, Mark D DeBoer University of Virginia, Charlottesville, VA; West Virginia University, Morgantown, WV; Virginia Commonwealth University, Richmond, VA; West Virginia University, Morgantown, WV Background:[br]The metabolic syndrome (MetS) is a cluster of clinical indices that increase risk for Type 2 diabetes (T2DM) and cardiovascular disease (CVD). The diagnosis of MetS is based on cut-off points for these different components, including waist circumference (WC), triglycerides (TG), HDL cholesterol, blood pressure (BP), and fasting glucose. However, the best way to diagnose MetS in children remains unclear, and current attempts result in ethnic discrepancies.[br]Methods:[br]Using 1999-2008 data from the National Health and Nutrition Examination Survey (NHANES), we designed a childhood continuous MetS (CC-MetS) risk score that is sex- and ethnicity-specific. We did so by performing confirmatory factor analysis of a single MetS factor, allowing the individual components (WC, TG, HDL, BP, glucose) to vary by sex and ethnicity when these components were significantly different by sex/ethnic group in their contribution to the MetS factor. The ethnic divisions were non-Hispanic white (NHW), non-Hispanic black (NHB) and Hispanic (Hisp). This provided a MetS score that is unique to each sex/ethnic group in the weighting of each component to the final continuous score. We then used ROC curves to compare this CC-MetS score to traditional ATP III-based MetS score (ATP-MetS), corresponding to a z-score cut off of 0.75. Using this cut-off for each of the race/ethnicities we then compared the ability of this score vs. ATP-MetS to predict elevations in surrogate markers of T2DM and CVD risk: insulin, high sensitivity C-reactive protein (hsCRP), and uric acid.[br]Results:[br]We generated sex- and ethnicity-specific equations that differed in the contribution of HDL and TG to the final CC-MetS score, as determined by the factor analysis. This score was significantly more sensitive than ATP-MetS score at detecting [ge]2 elevation among the surrogate markers: Males, ATP-MetS: NHW 56.3%, NHB 42.2%, Hisp 63.7%; CC-MetS: NHW 86.0%, NHB 75.6%, Hisp 75.6% (p[lt]0.05). Females, ATP-MetS: NHW 25.4%, NHB 23.9%, Hisp 38.3%; CC-MetS 47.1%, NHB 64.7%, Hisp 81.2% (p[lt]0.05).[br]Discussion:[br]These new equations for the CC-MetS score produce a clinically accessible and interpretable MetS score that is sex- and ethnicity-specific and that can be used to identify children at higher risk for developing adult diseases related to MetS, who could then be targeted for increased intervention. Additionally, they provide a powerful new outcome that can be utilized in childhood obesity and MetS research.[br][br]Sources of Research Support: NIH:. 1R21DK085363 (MDD and MJG). 5K08HD060739-02 (MDD).[br][br]Nothing to Disclose: MJG, CLI, SSS, MDD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 176 180 1214 SUN-157 PO10-01 Sunday 1009 2012


1008 ENDO12L_SUN-158 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) The Prognostic Impact of Fitness on Mortality in Diabetics with and without Left Ventricular Hypertrophy Khaled Alswat, Peter Kokkinos, Shikha Khosla, Raya Kheirbek, Eric Nylen Washington DC Veterans Affairs Medical Center (DCVAMC), George Washington University, Washington, DC; Washington DC Veterans Affairs Medical Center (DCVAMC), Georgetown University School of Medicine, Washington, DC; Washington DC Veterans Affairs Medical Center (DCVAMC), Washington, DC; Washington DC Veterans Affairs Medical Center (DCVAMC), Washington, DC [bold]Background:[/bold] Left ventricular hypertrophy (LVH) and type 2 diabetes mellitus are independent risk factors for mortality. Exercise capacity is inversely and independently associated with lower mortality risk in diabetics. However, the association between exercise capacity, LVH and mortality risk in diabetics has been poorly explored, and was therefore the aim of this study.[br][bold]Methods:[/bold] From 1986 to 2011, 866 men with type 2 diabetes (mean age 61[plusmn]10 years), underwent a standardized exercise stress test and echocardiographic evaluation at the Veterans Affairs Medical Center, Washington, DC. We established 2 fitness categories based on peak exercise capacity (METs) achieved. Individuals with a peak MET level within the lower 50th percentile ([lt]6 METs) were considered Low-Fit and those with peak MET level [ge]6 were Fit. Left ventricular mass (LVM) was calculated by a standardized formula and was then indexed to body size by dividing LVM by height in meters to the power of 2.7 to obtain LVM index. LVH was defined as LVM index [gt]48 g/m 2.7. To assess the risk associated by the interaction between fitness status and LVH we established four groups based on the fitness status of the individual and the presence or absence of LVH: Low-Fit/No LVH (n=225); Low-Fit/LVH (n=236); Fit/No LVH (n=218); and Fit/LVH (n=187). Low-Fit/No LVH was used as the reference groups for all Survival analyses.[br][bold]Results:[/bold] During the follow-up period of 24 years (median 8.9 years) there were 346 deaths for an annual mortality rate of 4.3% in the entire cohort. Cox proportional hazard analysis adjusted for age, BMI, hypertension, smoking and medications revealed 20% higher mortality risk for the Low-Fit/LVH group (HR: 1.20; CI: 0.93-1.56, P = 0.15). Mortality risk was 41% lower (HR: 0.59; CI: 0.42-0.82, P = 0.002) in the Fit/No LVH individuals and 43% lower mortality risk (HR: 0.57; CI: 0.40-0.81, P = 0.002) in the Fit/LVH individuals.[br][bold]Conclusions:[/bold] The presence of LVH in Low-fit individuals with type 2 diabetes was associated with a higher mortality risk. This risk was significantly decreased in individuals with moderate fitness regardless of LVH status.[br][br]Nothing to Disclose: KA, PK, SK, RK, EN 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 99 180 1215 SUN-158 PO10-01 Sunday 1010 2012


1009 ENDO12L_SUN-159 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Experimental Sleep Restriction Decreases Beta-Cell Function in Adults at Risk for Type 2 Diabetes Lindsay Bromley, John Booth III, Jennifer Kilkus, Luis Alcantar, Jacqueline Imperial, Plamen Penev University of Chicago, Chicago, IL; University of Chicago, Chicago, IL Short sleep ([lt]6 h/d) is associated with increased incidence of diabetes and sleep loss can interfere with insulin secretion and action. Adults with parental history of type 2 diabetes (T2DM) have a high risk of developing the disease, but the effects of experimental sleep restriction on their beta-cell function and systemic insulin resistance are not known. We examined the impact of experimental sleep restriction on insulin secretion and action in such individuals with increased metabolic risk.[br]Eleven healthy adults with parental history of T2DM (5F/6M, mean [SD] age 26 [3.0] y, BMI 23.5 [2.5] kg/m2) completed two inpatient studies including 8 nights with adequate vs. restricted time-in-bed (8.5 vs. 5.5 hours/day) in random crossover fashion 4 to 12 weeks apart. On two consecutive days at the end of each sleep condition, subjects underwent 3-hour hyperglycemic (8.5 vs. 5.5-h time-in-bed: plasma glucose 189 mg/dL [3] vs. 188 mg/dL [3], CV 5 [3] vs. 6 [2]) and euglycemic-hyperinsulinemic clamp testing (glucose 90 mg/dL [2] vs. 90 mg/dL [1], CV 5 [3] vs. 5 [2]; serum insulin 395 pmol/L [55] vs. 396 pmol/L [48]). Measures of insulin action included fasting HOMA-IR and the M-value during the last hour of the euglycemic clamp. Measures of beta-cell function included the area under the insulin curve between minutes 2-10 (1-st phase) and 120-180 (2-nd phase) of the hyperglycemic clamp. Repeated-measures analysis of variance was used to compare insulin secretion and action between the two sleep conditions.[br]Fasting blood glucose (82 mg/dL [3] vs. 85 mg/dL [5]; P=0.07) and insulin (21 pmol/L [3] vs. 27 pmol/L [4]; P=0.18) tended to be lower at the end of the 5.5-h time-in-bed condition, but this difference was not statistically significant. There was no difference in insulin sensitivity at the end of the 5.5 vs. 8.5-h time-in-bed condition (HOMA-IR 0.38 [0.05] vs. 0.50 [0.08]; M-value 6.3 [1.5] vs. 6.2 [2.0] mg/kg/min). Sleep restriction had no effect on first phase, but significantly decreased second phase insulin secretion (433 pmol/L [45] vs. 530 [46]; P=0.035) and the corresponding disposition index (P=0.040).[br]In conclusion, short-term experimental sleep restriction in at-risk adults with parental history of type 2 diabetes leads to reduced glucose-stimulated insulin secretion, but does not have a significant effect on systemic insulin resistance. Understanding the adaptive role and potential maladaptive consequences of this response requires further investigation.[br][br]Sources of Research Support: NIH grants R01-HL089637, CTSA-RR 024999 and P60-DK020595.[br][br]Nothing to Disclose: LB, JB, JK, LA, JI, PP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 523 180 1216 SUN-159 PO10-01 Sunday 1011 2012


1010 ENDO12L_SUN-160 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Losartan/Hydrochlorothiazide vs Losartan/Amlodopine in Hypertensive Patients with Metabolic Syndrome: The CATCH (Combination of Antihypertensive Treatment in Chiba) Study Ichiro Tatsuno, Takashi Terano Toho University Sakura Medical Center, Sakura, Japan; Chiba Aoba Municipal Hospital, Chiba, Japan [bold]Background:[/bold] The metabolic syndrome (Mets) has a high prevalence of hypertension with high cardiovascular risk, which sometime requires the combination therapy of anti-hypertensive drugs.[br][bold]Objective:[/bold] To compare the combination of losartan/hydrochlorothiazide with losartan/amlodopine in the treatment of Japanese hypertensive patients with Mets.[br][bold]Design:[/bold] 24-week, randomized, multicenter, open controlled study.[br][bold]Patients and methods:[/bold] We enrolled 65 hypertensive patients with Mets, who had a higher systolic pressure(SBP) of [ge]140 mm Hg and/or a higher diastolic pressure (DBP) of [ge]90 mm Hg under the treatment of an angiotensin-receptor blocker alone or a calcium channel blocker alone over 3 months, and randomized them to losartan 50mg/hydrochlorothiazide 12.5mg or losartan 50mg/amlodopine 5mg. If target blood pressure of [lt] 130/80 mm Hg was not reached 3 months after the treatment, losartan was doubled in both groups. Primary endpoint was SBP at 12-week, and the secondary endpoints were SBP, DBP (at 4, 8, and 24-week), adiponectin, 8-OhdG, HOMA-R, C-reactive protein (CRP), and albuminuria (at 12 and 24-week).[br][bold]Results:[/bold] Although both treatments of losartan/amlodopine and losartan/hydrochlorothiazide significantly improved SBP, the reduction rate of blood pressure with losartan/amlodopine (-20.8 [plusmn]13.53 mm Hg) was significantly higher than that with losartan/hydrochlorothiazide (-11.3 [plusmn]11.15 mm Hg) (P[lt]0.01). There were no significant difference between two groups in the secondary endpoints of adiponectin, 8-OhdG, HOMA-R, CRP, and albuminuria.[br][bold]Conclusion:[/bold] The combination therapy of losartan/amlodopine significantly lowered the SBP at 12-week in comparison with losartan/hydrochlorothiazide in hypertensive patients with metabolic syndrome.[br][br]Sources of Research Support: Clinical Trials Number: UMIN000001238.[br][br]Nothing to Disclose: IT, TT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 183 180 1217 SUN-160 PO10-01 Sunday 1012 2012


1011 ENDO12L_SUN-161 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Aromatase Inhibition in Post-Menopausal Breast Cancer Patients Is Associated with Increased Body Fat and Insulin Resistance: A Case-Control Study Fraser W Gibb, Ruth Andrew, Mike Dixon, Catriona Clarke, Brian R Walker University of Edinburgh, Edinburgh, UK; Western General Hospital, Edinburgh, UK; Western General Hospital, Edinburgh, UK BACKGROUND[br]Estrogen deficiency following menopause is implicated in adverse changes in body fat distribution and cardiometabolic risk factors. We hypothesized that the additional estrogen suppression effected by pharmacological aromatase inhibition exacerbates fat accumulation and insulin resistance. We previously demonstrated that anastrozole increases insulin resistance during hyperinsulinemic euglycemic clamps in a randomised crossover trial in healthy men. We have now tested this hypothesis in a [apos]real world[apos] clinical context, by studying breast cancer patients treated with aromatase inhibitors.[br]METHODS[br]20 post-menopausal breast cancer patients, who had received at least 1 year of aromatase inhibitor therapy (mean 27 months), were recruited with 20 age-matched healthy women. Insulin sensitivity was assessed by 75g OGTT with multiple measurements of glucose and insulin, yielding Matsuda insulin sensitivity index. Body fat was assessed by anthropometry and dual-energy x-ray absorptiometry (DEXA). BP and fasting plasma lipid profile were also assessed. Data are mean [plusmn] SEM, compared by Student[apos]s t-tests.[br]RESULTS[br]Cases and controls were matched for age (61.4[plusmn]1.4 versus 59.4[plusmn]1.1 years, p=0.26). Despite no differences in BMI (27.1[plusmn]0.9 versus 26.6[plusmn]1.0 kg/m2, p=0.68) or WHR, aromatase inhibitor treated women had higher body fat content (38.3%[plusmn]1.0 versus 34.7%[plusmn]1.3, p=0.03) and, as a result of a trend to higher insulin and no difference in glucose during OGTT, lower insulin sensitivity index (6.1[plusmn]0.5 versus 8.3[plusmn]0.6, p=0.03) than controls. No differences in lipid profile were observed but diastolic BP was 7mmHg higher in cases than controls (p=0.01).[br]DISCUSSION[br]Despite the limitations of case-control methodology, these findings in female patients corroborate our earlier observations in men in a rigorous randomised trial using gold-standard euglycemic hyperinsulinemic clamp assessments. Given the widespread use of aromatase inhibitors, prospective evaluation of their effects upon fat deposition and glucose metabolism, and the consequences for risk of diabetes and cardiovascular disease, is warranted.[br][br]Nothing to Disclose: FWG, RA, MD, CC, BRW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1261 180 1218 SUN-161 PO10-01 Sunday 1013 2012


1012 ENDO12L_SUN-162 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Testosterone Replacement Therapy in Hypogonadal Men Alters the HDL Proteome but Does Not Affect HDL Cholesterol Efflux Capacity Katya B Rubinow, Tomas Vaisar, Chongren Tang, Alvin M Matsumoto, Jay W Heinecke, Stephanie T Page University of Washington, Seattle, WA; VA Puget Sound Health Care System, Seattle, WA [bold]Introduction:[/bold] The effects of androgens on cardiovascular disease (CVD) risk in men remain unclear. Exogenous testosterone administration can decrease serum concentrations of high density lipoprotein-associated cholesterol (HDL-C), but low circulating androgen levels have been associated with increased CVD and CVD-related mortality in men. Recently, clinical studies have demonstrated that both HDL protein cargo and cholesterol efflux capacity exhibit significant correlations with coronary artery disease. To better characterize the relationship between androgens and HDL, we investigated the effects of testosterone replacement in hypogonadal men on HDL protein composition and HDL-mediated cholesterol efflux.[br][bold]Methods:[/bold] 23 older, hypogonadal men (ages 51-83, baseline testosterone [lt]280 ng/dL) were administered replacement testosterone therapy (1% transdermal gel) to achieve physiologic serum levels (500-1000 ng/dL) with or without the 5[alpha]-reductase inhibitor dutasteride. At baseline and after 3 months of testosterone replacement, fasting lipids were measured, the protein cargo of HDL was determined, and the cholesterol efflux capacity of serum HDL was assessed.[br][bold]Results:[/bold] Testosterone replacement did not affect HDL-C concentrations but conferred significant changes in the HDL proteome, with an increase in HDL-associated fibrinogen alpha chain (FGA) and a decrease in apolipoprotein A-IV (apoA-IV) evident after 3 months of treatment (p=0.023 and p=0.016, respectively, versus baseline). Exogenous testosterone did not alter the cholesterol efflux capacity of serum HDL. No differences were observed between men who received testosterone alone and those who received testosterone with dutasteride.[br][bold]Conclusions:[/bold] Testosterone replacement in older, hypogonadal men alters the protein composition of HDL but does not significantly change serum HDL-mediated cholesterol efflux. These effects appear independent of testosterone conversion to dihydrotestosterone. Further research is needed to determine how changes in HDL protein content affect HDL function and CVD risk in men.[br][br]Sources of Research Support: National Institute of Aging grant RO1AG037603; the Eunice Kennedy Shriver National Institute of Child Health and Human Development cooperative agreement U54 HD42454 as part of the Cooperative Contraceptive Research Centers Program; the Pacific Northwest Prostate Cancer SPORE (P50CA097186); the Mass Spectrometry Core, supported by the Diabetes and Endocrinology Research Center (University of Washington) P30 DK017047; a Pilot and Feasibility Award from the Diabetes and Endocrinology Research Center and the University of Washington (grant DK017047 from the National Institute of Diabetes and Digestive and Kidney Diseases); the National Institute of Diabetes, Digestive, and Kidney Diseases grant T32DK007247; the University of Washington Nutrition and Obesity Research Center (NIH grant P30DK035816); the American Heart Association grant 0830231N; the National Heart, Lung, and Blood Institute grants P01HL112625, R01HL086798, and P01HL092969.[br][br]Disclosures: AMM: Principal Investigator, Abbott Laboratories, GlaxoSmithKline, Ascend; Consultant, Novartis Pharmaceuticals. STP: Principal Investigator, Abbott Laboratories. Nothing to Disclose: KBR, TV, CT, JWH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 227 180 1219 SUN-162 PO10-01 Sunday 1014 2012


1013 ENDO12L_SUN-163 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) The Effect of Carvedilol and Salsalate in the Prevention of Free Fatty Acid-Induced Hypertension, Endothelial Dysfunction, and Inflammation in Overweight Healthy Subjects Francisco J Pasquel, Aidar Gosmanov, Farnoosh Farrokhi, Joselita Siqueira, Christopher Newton, Limin Peng, Ibhar Al Mheid, Gonzalo Robalino, Dawn Smiley, Guillermo E Umpierrez Emory University School of Medicine, Atlanta, GA; University of Tennesee Health Science Center, Memphis, TN; Rollins School of Public Health, Atlanta, GA Extensive evidence indicates that Intralipid infusion leading to supra-physiologic free fatty acids (FFAs) levels is associated with HTN, endothelial dysfunction and inflammatory response. As an attempt to elucidate underlying mechanisms, we determined the effect of 6 week salsalate and carvedilol treatment on the prevention of FFA-induced hypertension, endothelial dysfunction, and inflammation in overweight healthy subjects.[br]Methods: In this prospective randomized double-blind placebo-controlled study a total of 35 overweight/obese healthy subjects received placebo(n=12, age 37.5[plusmn]8 yrs, BMI 29[plusmn]4 kg/m2, systolic blood pressure (SBP: 115[plusmn] 11 mm Hg), salsalate(n=11, age 34[plusmn] 11 yrs, BMI 35[plusmn]9kg/m2, SBP 111[plusmn] 16mmHg), or carvedilol(n=12, age 41[plusmn]11 yrs, BMI 30[plusmn]6 kg/m2, SBP 111[plusmn] 13 mmHg) for 6 weeks. All patients received a 24-h Intralipid 20% infusion at 20 mL/h both at baseline and after 6 weeks of therapy. BP was measured every 4 hours, flow mediated dilation (FMD) was evaluated at 4 and 24 hours, and inflammation markers were measured every 6 hours during the infusion.[br]Results: FFA levels increased similarly during Intralipid infusion at baseline and after the 6 week of therapy in all groups (p[pound]0.01). There were no differences in FFA concentration during Intralipid infusion at baseline and follow-up studies among groups. Systolic BP increased during Intralipid infusion in all groups at baseline(all P[lt]0.05); however, neither salsalate or carvedilol treatment did prevent the lipid-induced rise in BP after 6 weeks of therapy.[br]Intralipid infusion was associated with a significantly elevation of TNF-alpha and IL-6 at 12 h, 18h, and 24 h during baseline infusion. Six weeks treatment with salsalate, carvedilol or placebo did not affect the inflammatory cytokine surge during Intralipid infusion. Low-dose Intralipid infusion was not associated with significant changes in FMD from baseline. In addition, the administration of salsalate, carvedilol and placebo was not associated with significant changes in endothelial function.[br]In summary, physiological increases in FFA levels during low-dose Intralipid infusion were associated with an increase in systolic BP and inflammatory markers, but not with impaired endothelial function in overweight/obese healthy subjects. Treatment with carvedilol or salsalate for 6 weeks did not prevent Intralipid-induced increase in blood pressure, inflammatory response or changes in endothelial function in obese healthy subjects.[br][br]Nothing to Disclose: FJP, AG, FF, JS, CN, LP, IAM, GR, DS, GEU 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2136 180 1220 SUN-163 PO10-01 Sunday 1015 2012


1014 ENDO12L_SUN-164 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Cardiovascular Outcomes in Turner Syndrome: Insights from a French Cohort Bruno Donadille, Alexandra Rousseau, Delphine Zenaty, Sylvie Cabrol, Carine Courtillot, Dinane Samara, Sylvie Salenave, Philippe Bouchard, Philippe Chanson, Michel Polak, Philippe Touraine, Yves Lebouc, Jean-Claude Carel, Juliane Leger, Sophie Christin-Maitre, Heart [amp] Turner Study Group Centre de Reference des Maladies Endocrines Rares de la Croissance, H[ocirc]pital St Antoine, Paris, France; Assistance Publique [ndash] H[ocirc]pitaux de Paris, Universit[eacute] Pierre et Marie Curie, Paris, France; Centre de Reference des Maladies Endocrines Rares de la Croissance, H[ocirc]pital Robert Debr[eacute], Paris, France; Centre de Reference des Maladies Endocrines Rares de la Croissance, H[ocirc]pital Trousseau, Paris, France; Centre de Reference des Maladies Endocrines Rares de la Croissance, H[ocirc]pital Piti[eacute]-Salp[eacute]tri[egrave]re, Paris, France; Centre de Reference des Maladies Endocrines Rares de la Croissance, H[ocirc]pital Necker, Paris, France; Centre de Reference des Maladies Endocrines Rares de la Croissance, H[ocirc]pital Bic[ecirc]tre, Paris, France OBJECTIVE: Congenital cardiovascular malformations and aortic dilatation (AD) are frequent in patients with Turner syndrome (TS). The objective of this study was to investigate cardio-aortic outcomes in a large cohort of patients, including children and adults.[br]METHODS: We recruited 336 patients with TS from a network of tertiary centers. We reviewed their files, checking for cardiovascular events, cardiac valve abnormalities, ventricular thickness and aortic diameters indexed to body surface area (BSA) from MRI (n=110) or echocardiography (n=300).[br]RESULTS: Informative cardiovascular data were available for 233 patients. Vascular surgery was reported in 7.4% of the cohort, at a median age of 1 year (range: 0-48). The first cause of surgery was aortic coarctation, detected in 6.9% at a median age of 9.5 years (range: 0-60). Aortic bicuspid valve (BAV) was detected in 21% at a median age of 20 [25th-75th percentiles: 15-30]. At least one aortic diameter exceeded 32 mm in 12% of the cohort. It was detected at a median age of 19 [7-30]. When indexed to BSA, at least one aortic diameter exceeded 20 mm/m2 in 39% of the cohort. A 45,X karyotype was significantly associated with AD (p [lt] 9.10-3) and BAV (p=0.007).[br]CONCLUSION: Our study shows that cardiovascular monitoring for Turner syndrome patients is currently insufficient. BAV is present at birth, but often remains undiagnosed until later in life. Aortic dilatation is more frequent than previously reported. Therefore careful cardiovascular monitoring, with indexing to BSA, is thus required throughout the patient[apos]s life.[br][br]Nothing to Disclose: BD, AR, DZ, SC, CC, DS, SS, PB, PC, MP, PT, YL, J-CC, JL, SC-M 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 702 180 1221 SUN-164 PO10-01 Sunday 1016 2012


1015 ENDO12L_SUN-165 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) The Genetic Association of the Adiponectin Gene Variant rs2241766 [+45T/G] and rs1501299 (276G[gt]T) with Angiographic Finding, Diabetes and Adiponectin among Acute Coronary Syndrome Patients Nasser M Rizk, Isra Meri, Ayman El -Menyar, Jassim Al Suwaidi Qatar University, Doha, Qatar; Hammad Medical Corporation-Hospital, Doha, Qatar Background: Adiponectin is a fat derived hormone, known to decrease in type 2 diabetes (DM) and acute coronary syndrome (ACS). Two single nucleotide polymorphisms (SNPs) (rs1501299 and rs2241766) in ADIPOQ gene and their possible association with ACS were investigated among Arab population in Qatar.[br]Methods: A case-control association study was performed on 71 ACS Arab patients with diabetes (case) and 71 ACS without diabetes (control), from Qatar. Genotypes were determined using TaqMan real time PCR assay. Serum adiponectin level was determined using ELISA.[br]Results: The genotype distribution of the rs2241766 based upon the angiographic findings for GG, GT and TT were (57.0%, 38.0%, 5.0% vs. 63.0%, 33.0%, 4.0%, vs.68.0%, 28.0%, 4.0%, p=0.813) for the single, double and three vessels affected, respectively. In addition, there was no significant association between the number of the vessels and the genotype distribution for the rs1501299, and the genotype frequencies for GG, GT and TT were (47.8%, 30.1%, 13.1% vs. 41.6%, 54.2%, 4.2%, and 48.0%, 44.0% and 8.0%, p=0.589), for the single, double and three vessels affected, respectively. All allele frequencies were in HWE equilibrium, P [gt]0.05. No significant association between genotype distribution and diabetic state for both polymorphisms was detected. GG, GT and TT genotype frequencies of the rs2241766 and for rs12255372 variant showed (52.9%, 25.7%, 21.4%, vs. 36.8%, 29.4%, 33.8% p=0.128), and (47.1%, 42.6%, 10.3%, vs. 35.7%, 48.6%, 15.7% p=0.583) among diabetic and non-diabetic, respectively. Mean values and SEM of total adiponectin level ([micro]g/ml) were; 13.3[plusmn]1.9, 8.13[plusmn]1.0 and 8.9 [plusmn]1.4 for the single, double and three vessels affected, p value= 0.082 and was 8.3[plusmn]0.91, and 7.64[plusmn]0.82, p=0.542 among diabetic and non-diabetic, respectively. In contrast, adiponectin level ([micro]g/ml) was significantly associated with genotype distribution among diabetic and non-diabetic subjects for rs2241766, only. The mean values and SEM of total adiponectin level ([micro]g/ml) among non-diabetic and diabetic were (11.4[plusmn]1.45, 8.6[plusmn]1.2, 4.8[plusmn]1.2, p=0.002 and 9.6[plusmn]1.1, 6.8[plusmn]1.1 and 3.4[plusmn]0.6, p= 0.006) an (10.2[plusmn]1.3, 8.2[plusmn]1.2, 5.4[plusmn]1.6, p=0191, a 7.3[plusmn]1.1, 7.4[plusmn]0.9 and 10.8[plusmn]1.1, p=0.639) by GG, GT and TT distribution for rs2241766 and rs1501299, respectively.[br]Conclusion: This study suggests that carriers of T allele of rs2241766 SNP+45T[gt]G in the ADIPO gene is characterized by low adiponectin level in ACS patients among Arabs populations in Qatar.[br][br]Sources of Research Support: Qatar National Research Fund, UREP Fund # 4-3-42.[br][br]Nothing to Disclose: NMR, IM, AE-M, JAS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2332 180 1222 SUN-165 PO10-01 Sunday 1017 2012


1016 ENDO12L_SUN-166 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) HbA1c Is Associated with Dyslipidemia in an Apparently Healthy Older Nondiabetic Adults Jin Hwa Kim, Ji In Kang, Sang Yong Kim, Hak Yeon Bae Chosun University Hospital, Gwangju, Republic of Korea [bold]Objective: [/bold]Elevated HbA1c levels are associated with a higher incidence of microvascular and macrovascular complications in diabetes. Data from the National Health and Nutrition Examination Survey and the Framingham Offspring Study suggest that HbA1c levels increase with age even after adjustments for demographic and glycemic variables among nondiabetic subjects. The aim of this study is to estimate the association of HbA1c with cardiovascular disease risk factors in an apparently healthy older nondiabetic adults.[br][bold]Methods: [/bold]We retrospectively studied asymptomatic Korean adults of 60 years old or older who underwent voluntary regular health check-ups at the Health Promotion Center of Chosun University Hospital (Gwangju, Republic of Korea) from January 2009 to September 2011. Individuals previously diagnosed with diabetes, hypertension, or dyslipidemia and those treated with hypoglycemic agents, antihypertensive agents, and lipid lowering agents were excluded. After these exclusions, 421 individuals were included in this study.[br][bold]Results: [/bold]HbA1c revealed a significant correlation with age, waist circumference, fasting plasma glucose, triglyceride, LDL cholesterol, and HDL cholesterol. Multiple regression analysis showed that HbA1c had a association with fasting plasma glucose([beta] [plusmn] SE, 0.009 [plusmn] 0.002), LDL cholesterol (0.001 [plusmn] 0.001), and HDL cholesterol (-0.003 [plusmn] 0.001).[br][bold]Conclusion:[/bold] The HbA1c level is independently correlated with dyslipidemia other than fasting glucose level in older nondiabetic Korean adults.[br][br]Nothing to Disclose: JHK, JIK, SYK, HYB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 270 180 1223 SUN-166 PO10-01 Sunday 1018 2012


1017 ENDO12L_SUN-167 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Pulse-Wave Velocity Is Associated with Hemoglobin A1c in Diabetic Patients with Nephropathy Martin M Sarkar, Samip J Parikh, Jigar Bhagatwala, Ishita Kotak, Haidong Zhu, Yanbin Dong Georgia Health Sciences University, Augusta, GA; Georgia Health Sciences University, Augusta, GA; Georgia Health Sciences University, Augusta, GA; Georgia Health Sciences University, Augusta, GA [bold]Background:[/bold] Pulse wave velocity (PWV) is well-recognized as a noninvasive marker of arterial stiffness. The purpose of this study is to evaluate the relationship of PWV parameters with traditional cardiovascular risk factors such as adiposity [body mass index (BMI), percent body fat (%BF), and total fat mass], blood pressures [systolic blood pressure (SBP) and diastolic blood pressure (DBP)], heart rate (HR), urine albumin to creatinine ratio (UACR), hemoglobin A1C (HbA1C) and lipid profiles in patient with type 2 diabetes. [bold]Methods: [/bold]We recruited a total of 36 (mean age 53 [plusmn] 10 years, 58% females, 75% African Americans) type 2 diabetic patients with preserved renal function (calculated eGFR [gt] 60 ml/min/1.73 m[sup]2[/sup]). Adiposity was assessed by using whole body dual-energy x-ray absorptiometry (DXA). Carotid-femoral and carotid-radial PWV were measured noninvasively with applanation tonometry (SphygmoCor). [bold]Results:[/bold] In the entire cohort, neither carotid-femoral nor carotid-radial PWV was correlated with BMI, %BF, total fat mass, SBP, DBP, HR, UACR, HbA1C, LDL, and HDL. However, in diabetic patients with nephropathy (UACR [gt] 30 [micro]g/g, N=10), carotid-femoral PWV was found to be positively associated with HbA1C (r = 0.82, p = 0.02). The relationship remained significant even after adjusting for age, race, and sex. [bold]Conclusion:[/bold] Assessment of arterial stiffness by PWV parameters did not appear to be correlated with traditional cardiovascular risk factors in the entire cohort as originally hypothesized. However, in diabetic patients with nephropathy, carotid-femoral PWV was positively associated with HbA1C. Poorer glycemic control is a well-known risk factor for the progression of diabetic nephropathy, and possibly the development of vascular complications in diabetic patients with nephropathy. Further studies with large sample size are on the way to validate the usefulness and predictable value of PWV in this population.[br][br]Nothing to Disclose: MMS, SJP, JB, IK, HZ, YD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1602 180 1224 SUN-167 PO10-01 Sunday 1019 2012


1018 ENDO12L_SUN-168 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Low Femoral Neck Bone Mineral Density Is Related to Carotid Artery Intima-Media Thickness in Men with Type 2 Diabetes Mellitus Mario Almeida Coutinho, Nara Crispim Carvalho, Paula Prazeres Aragao, Francisco Alfredo Bandeira Agamenon Magalh[atilde]es Hospital, University of Pernambuco Medical School, Recife, Brazil; Agamenon Magalh[atilde]es Hospital, University of Pernambuco Medical School, Recife, Brazil; Magalh[atilde]es Hospital, University of Pernambuco Medical School, Recife, Brazil [bold]BACKGROUND:[/bold] Osteoporosis and atherosclerosis share common risk factors and the association of low bone mass with increased cardiovascular morbidity and mortality has been demonstrated in some studies. Nevertheless, most studies have been focused on women and only a few on individuals with type 2 diabetes mellitus (T2DM). [bold]OBJECTIVES:[/bold] To assess the association between bone mineral density and intima-media thickness of the carotid (CIMT) artery by doppler ultrasonography in men with T2DM. [bold]STUDY DESIGN AND METHODS: [/bold]We studied 24 men with T2DM, aged 61 years (SD [plusmn] 6.4), 75% with more than five years since the diagnosis of diabetes, 41.6% were on statin, 4.1% were smokers and with body mass index (BMI) of 28.1 kg/m2 (SD [plusmn] 3.4), abdominal circumference (AC) of 97.8cm (SD [plusmn] 8.4), systolic blood pressure (SBP) of 143.8mmHG (SD [plusmn] 18.3), diastolic blood pressure (DBP) of 85.8 mmHg (SD [plusmn] 12.3), HbA1C of 7.5% (SD [plusmn] 1.3), triglycerides of 141.7 mg/dL (SD [plusmn] 73), LDL-cholesterol of 103.3 mg/dL (SD [plusmn] 35.9) and HDL-cholesterol of 41.6 mg/dL (SD [plusmn] 11.6). They were stratified into groups according to bone mineral density (T score [lt] -2.5 and -2.5 a -1) by dual-photon X-ray absorption absorptiometry (GE Co., Waltham, Massachusets). CIMT was calculated using an average of three measurements at the common terminal carotid artery on bilateral bifurcation by Doppler ultrasonography (USG MODE LOGIC-B-E-GE Medical Systems). [bold]RESULTS:[/bold] There were no significant differences between groups regarding age, duration of T2DM, BMI, AC, SBP, DBP, statin use, smoking, HbA1C, cholesterol or triglycerides. For the femoral neck (FN), the group with normal bone mineral density (BMD) had a mean CIMT of 0.7 mm and the group with osteopenia or osteoporosis had a CIMT of 0.86 mm (p = 0.007). As for the lumbar spine, the group with normal BMD had mean measurements of CIMT of 0.81 mm and the group with osteopenia or osteoporosis had an average of 0.8 mm (p = 0.98). [bold]CONCLUSION:[/bold] Our data demonstrate a negative association between bone mineral density and carotid intima-media thickness in type 2 diabetic men, which was unrelated to the traditional risk factors for atherosclerotic disease and degree of diabetes control.[br][br]Nothing to Disclose: MAC, NCC, PPA, FAB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1314 180 1225 SUN-168 PO10-01 Sunday 1020 2012


1019 ENDO12L_SUN-169 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Carotid Intima-Media Thickness in Type 2 Diabetes Mellitus with and without Ischemic Stroke Sunil K Kota, Siva K Kota, Svs Krishna, Lalit K Meher, Kirtikumar D Modi Medwin Hospital, Hyderabad, India; Central Security Hospital, Riyadh, Saudi Arabia; MKCG Medical College, Berhampur, India Objective:To find cut off point for carotid intima media thickness (CIMT) for ischemic stroke in type 2 diabetes mellitus(T2DM).[br]Methods:80 subjects with age 30-75 years(M:F=57:23)were divided into 3 groups: A)T2DM subjects with ischemic stroke,B)T2DM subjects and C)healthy subjects.Patients with cardioembolic stroke,hemorrhagic stroke and stroke secondary to trauma,impaired coagulation or tumor were excluded.All subjects underwent B-mode ultrasonography of common carotid arteries to determine CIMT along with history taking, physical examination and routine laboratory investigations including included FBS, PPBS, HbA1C, renal profile, lipid profile and microalbuminuria.[br]Results:40 patients(M:F=30:10) were in group A with mean age 60.4[plusmn]10.2 years.Group B had 20 patients(M:F=12:8) with mean age 56.8[plusmn]11.7 years.Group C had 20 subjects (M:F=15:5) with mean age 51.3[plusmn]16.7 years.CIMT [gt]0.8 mm was associated with occurrence of stroke in group A subjects.Patients with T2DM with or without ischemic stroke were found to have significantly higher prevalence of increased CIMT (92.5 % in group A, 80% in group B and 20% in group C).The mean CIMT of study population was 0.84 [plusmn] 0.2 mm (Group A-1.06[plusmn]0.2mm,Group B- 0.97[plusmn]0.26 mm and Group C- 0.73[plusmn]0.08 mm).The mean CIMT was not significantly different between T2DM patients with or without ischemic stroke (p-0.08). Type I variety (plaque with thin rim over the surface, but predominantly anechoic) was the most common among subjects with T2DM and ischemic stroke.However,the mean CIMT was significantly higher in diabetic subjects with and without stroke compared to healthy subjects(group A versus group C:p-0.003, group B versus group C:p-0.03, combined group A [amp] B versus group C: p-0.006).Higher age,smoking,hypertension,hyperlipidemia,low HDL cholesterol,glycemic parameters,microalbuminuria and duration of diabetes were independently and significantly related to CIMT.[br]Discussion: CIMT is used as a noninvasive tool for assessment of atherosclerosis.Rotterdam study has demonstrated CIMT [gt] 1.2 mm as a risk for stroke in middle aged adults(1).CIMT is demonstrated as a surrogate marker for subclinical coronary artery disease (CAD) in diabetic patients.Our study highlights CIMT [gt] 0.8 mm as a risk factor for occurrence of stroke in T2DM.T2DM patients with or without stroke are at equal risk for development of stroke,as evidenced by presence of CIMT [gt] 0.8 mm.[br]Conclusion:CIMT[gt]0.8 mm is a surrogate risk marker ischemic stroke in T2DM patients.[br][br]1. Hoskote SS, et al., J Assoc Physians India. 2008. 56: 577.[br][br]Nothing to Disclose: SKK, SKK, SK, LKM, KDM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1396 180 1226 SUN-169 PO10-01 Sunday 1021 2012


1020 ENDO12L_SUN-170 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Increased Intima-Media Thickness and Central Blood Pressure in Hypertensive Individuals with Cognitive Impairment Jose Fernando Vilela-Martin, Luciana Neves Cosenso-Martin, Eros Mota-Dias, Luiz Tadeu Giollo-Junior, Carolina Neves Cosenso-Sacomani, Juan Carlos Yugar-Toledo, Heitor Moreno-Junior State Medical School of Sao Jose do Rio Preto (FAMERP), Sao Jose do Rio Preto, Brazil; Faculty of Medical Sciences, University of Campinas, Campinas, Brazil Introduction: The role of hypertension in the loss of cognitive function is controversial. Relationships of hypertension with increases in cerebral vascular resistance, diffused lesions and multiple lacunar infarcts of the white matter are well known. Thus, the objectives of this study were: To evaluate the relationship between hypertension and cognitive dysfunction (CD), identify risk factors and determine the association between markers of early vascular disease and CD in hypertensive individuals.[br]Methods: Two hundred individuals aged between 40 and 80 years old were evaluated in this cross sectional study. Fifty participants were normotensive (NT). The remaining 150 hypertensive patients were subdivided into two groups, those with CD (HCD) and those without CD (HNCD). All participants underwent clinical evaluations and biochemical blood tests were performed. CD was investigated using the Mini Mental State Examination (MMSE) following the guidelines for its use in Brazil. The impact of hypertension on the arterial bed was assessed by identifying and measuring changes in the intima-media thickness (IMT) by vascular ultrasonography of the carotid arteries and analyses of the central blood pressure and Augmentation Index by applanation tonometry of the radial artery.[br]Results: There were no significant differences in the plasma concentrations of total cholesterol, high-density lipoprotein cholesterol and triglycerides of the three groups. The serum creatinine and estimated glomerular filtration rate were within normal ranges for all three groups. A significantly lower MMSE score was recorded for the HCD Group compared to the HNCD and NT Groups (p[lt]0.05).[br]The IMT was significantly different between the HNCD and HCD Groups (p=0.0124). A significant difference in the IMT was also observed between hypertensive patients and the NT Group (p[lt]0.0001).[br]The central systolic pressure was significantly higher in the HCD and HNCD Groups compared to NT Group (p[lt]0.0001). There were no significant differences in the Augmentation Index (corrected for heart rate) between the three groups (HCD, HNCD and NT).[br]Conclusions: Hypertensive patients with CD have changes in the vascular morphology characterized by an increased carotid IMT and hemodynamic functional impairment manifested by elevated central systolic blood pressure.[br][br]Nothing to Disclose: JFV-M, LNC-M, EM-D, LTG-J, CNC-S, JCY-T, HM-J 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1046 180 1227 SUN-170 PO10-01 Sunday 1022 2012


1021 ENDO12L_SUN-171 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Prevalence of Subclinical Atherosclerosis and Its Associated Factors in Hyperlipidemic Korean Adults with Diabetes (ALTO Study) Eun Gyoung Hong, Hyuk Sang Kwon, Sin Gon Kim, Dong Jun Kim, Seung Jin Lee, Hae Rung Song, Dong Sun Kim Hallym University College of Medicine, Seoul, Republic of Korea; Catholic University College of Medicine, Seoul, Republic of Korea; Korea University College of Medicine, Seoul, Republic of Korea; Inje University College of Medicine, Seoul, Republic of Korea; AstraZeneca, Seoul, Republic of Korea; Hanyang University College of Medicine, Seoul, Republic of Korea [bold]Background: [/bold]Cardiovascular disease (CVD) and atherosclerotic diseases frequently occur in diabetic patients. An increased carotid intima-media thickness (cIMT) and the presence of atherogenic plaque, reflecting atherosclerosis, may predict coronary heart disease (CHD) and stroke after adjustment for other risk factors. Dysfunction of the vascular endothelium and chronic low-grade inflammation are key features of the atherosclerosis, both of which are often observed among diabetic patients. Data which show distribution of cIMT, presence of plaque, and associated factors among Korean diabetic patients are limited. We investigated the prevalence of subclinical atherosclerosis and atherogenic plaque, defined by the mean cIMT, and associated factors among hyperlipidemic Korean adults with diabetes but without established CVD.[br][bold]Methods:[/bold] A multicenter, cross-sectional study was performed in 189 Korean diabetic patients without established or previous CVD. The cIMT was measured with 3D ultrasonographic examinations by a skilled technician. Anthropometric and serum cardiometabolic profiles were measured. The mean cIMT in the healthy Korean population was used as the cut-off value for atherosclerosis.[br][bold]Results:[/bold] The prevalence of subclinical atherosclerosis was 20.2%. Atherogenic plaque was found in 59% and 54% of right and left carotid arteries, respectively. The mean cIMT was 0.61[plusmn]0.19 mm in the right and 0.62[plusmn]0.2 mm in the left artery. The mean cIMT was significantly correlated with age (r=0.31, p=0.0001), duration of diabetes (r=0.19, p=0.0146), Log high-sensitivity C-reactive protein (hs-CRP) (r=0.22, p=0.0031), and ApoAI (r=-0.17, p=0.0281). In multiple linear regression models, mean cIMT was positively associated with age (b=0.004, p=0.0023) and Log hs-CRP (b=0.035, p=0.0069), whereas ApoAI showed a negative association with mean cIMT (b=-0.001, p=0.0277). Subjects with additional risk factors for CHD showed significantly higher mean cIMT values than those with diabetes only (0.62[plusmn]0.19 vs 0.54[plusmn]0.09 mm, p=0.0375). The odds ratio for presence of atherogenic plaque of the carotid artery according to age was 1.1 (95% CI: 1.04, 1.16) in men, 1.09 (95% CI: 1.02, 1.16) in women.[br][bold]Conclusion:[/bold] Hyperlipidemic Korean adults with diabetes showed a high prevalence of subclinical atherosclerosis and atherogenic plaque of the carotid artery. Age, hs-CRP, and ApoAI were the main predictors of the mean cIMT among hyperlipidemic Korean adults with diabetes.[br][br]Nothing to Disclose: EGH, HSK, SGK, DK, SJL, HRS, DK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1391 180 1228 SUN-171 PO10-01 Sunday 1023 2012


1022 ENDO12L_SUN-172 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) A Possible Role of Undercarboxylated Osteocalcin in Cardiovascular Risk Assessment in Patients with Type 2 Diabetes Assim A Alfadda, Afshan Masood, Hafedh Dekhil, Muhammad Azhar Chishti, Shaffi Ahamed Shaikh College of Medicine, King Saud University, Riyadh, Saudi Arabia; College of Medicine, King Saud University, Riyadh, Saudi Arabia; College of Medicine, King Saud University, Riyadh, Saudi Arabia; College of Medicine, King Saud University, Riyadh, Saudi Arabia Background[br]Studies have shown that total osteocalcin (TOC) is associated with metabolic syndrome and therefore might influence the risk of cardiovascular disease in humans. Undercarboxylated osteocalcin (UOC) regulates insulin secretion and sensitivity in mice but its relation with metabolic syndrome in humans has not been studied.[br]Objectives[br]To determine whether UOC is related to metabolic syndrome and its components in patients with type 2 diabetes mellitus (T2DM).[br]Materials and Methods[br]A cross-sectional study of 203 patients with T2DM (mean age 52 years; 40% females) with and without metabolic syndrome was conducted at the Obesity Research Center, King Saud University, Riyadh. Metabolic syndrome was defined according to NCEP-ATP III criteria. Stepwise multiple regression analysis was performed between the three outcome variables: (i)TOC (ii)UOC (iii) carboxylated osteocalcin (COC) and metabolic syndrome components, HbA1c and lipid profile.[br]Results[br]In the whole sample studied, TOC was negatively correlated with BMI (r = - 0.153, p = 0.035) and HbA1c (r = - 0.211, p = 0.006), but not with HDL-cholesterol (r = 0.029, p = 0.693) and ApoA-1(r = - 0.026, p = 0.717). UOC was positively correlated with HDL-cholesterol (r = 0.172, p = 0.016) and ApoA-1(r = 0.158, p = 0.026). COC was not associated with any of the parameters studied. Both TOC and UOC were lower in patients with metabolic syndrome compared to those without metabolic syndrome (9.24 [plusmn] 3.9 [italic]vs.[/italic]11.01 [plusmn] 4.9 [mu]g/l (p = 0.007), and 1.41 [plusmn] 1.13 [italic]vs.[/italic] 1.79 [plusmn] 1.46 [mu]g/l (p = 0.038), respectively). In patients with metabolic syndrome, UOC was also significantly correlated with both HDL-cholesterol (r = 0.246, p = 0.007) and ApoA-1(r = 0.237, p = 0.009). Stepwise multiple regression analysis has shown that UOC was independently related to HDL-cholesterol (b = 0.43, r[sup]2[/sup] = 0.06, p = 0.013), while TOC was independently related to triglycerides (b = - 1.041,r[sup]2[/sup] = 0.04,p = 0.049).[br]Conclusions[br]Lower levels of UOC in patients with metabolic syndrome compared to those without metabolic syndrome and its association with HDL-cholesterol and ApoA-1suggest a possible role of UOC in cardiovascular risk assessment in patients with T2DM. Further studies are required to determine whether interventions which raise specifically levels of UOC would decrease cardiovascular risk.[br][br]Sources of Research Support: This work was supported by a grant from the National Plan for Science and Technology, King Saud University and King Abdulaziz City for Science and Technology, Riyadh, Saudi Arabia (grant no. 08-MED513-02).[br][br]Nothing to Disclose: AAA, AM, HD, MAC, SAS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 455 180 1229 SUN-172 PO10-01 Sunday 1024 2012


1023 ENDO12L_SUN-173 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Aspirin Resistance in Young Men with Type 2 Diabetes Subhashini Yaturu, Emmy Dier, Shaker Mousa Stratton VA Medical Center, Albany, NY; Pharmacy Research Institute, Rensselaer, NY Cardiovascular events are the leading causes of death in subjects with diabetes. ADA recommends low dose aspirin for primary and secondary prevention of ischemic events in patients with diabetes. Aspirin resistance (AR) has emerged as a new concept to account for some of the treatment failures. AR involves inadequate or lack of inhibition of the cyclooxygenase-1-mediated thromboxane A[sub]2[/sub] pathway.The aim of the study was to evaluate aspirin resistance (AR) in young men with type 2 diabetes and evaluate correlation with clinical risk factors and adipokines. Aspirin resistance was measured by urinary 11-dehydro-thromboxane beta-2 (11DhTx2) concentrations with an enzyme immunoassay kit. One hundred and forty two subjects with type 2 diabetes had 11DhTx2 measurement in the urine samples. Levels of 11DhTx2, a major urinary metabolite of thromboxane at [ge] 1500 pg/mg of creatinine is considered as AR. Levels of 11DhTx2 correlate with duration of diabetes ([italic]r [/italic]= 0.12;[italic] p[/italic][lt]0.05), presence of micro albumin and IL-6 ([italic]r[/italic]= 0.12;[italic] p[/italic][lt]0.05); negative correlation with systolic and diastolic blood pressure, and waist circumference ([italic]r[/italic]= 0.12;[italic] p[/italic][lt]0.05). Levels of 11DhTx2 did not correlate with insulin levels, insulin resistance or other inflammatory markers such as CRP, TNF alpha. Approximately 53% patients were resistant to aspirin. Duration of diabetes and micro albumin levels had a direct influence on the aspirin resistance.[br][br]Sources of Research Support: Part of the work supported by VA Merit review Grant.[br][br]Nothing to Disclose: SY, ED, SM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 724 180 1230 SUN-173 PO10-01 Sunday 1025 2012


1024 ENDO12L_SUN-174 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Exploration of the Interaction between Diabetes, Cardiovascular Disease, Depression, and Dementia in the Community-Dwelling Medicare Beneficiary Population Elizabeth Ann Koller, Gerald Stanton Adler Centers for Medicare and Medicaid Services, Baltimore, MD; Centers for Medicare and Medicaid Services, Baltimore, MD [bold]Background:[/bold] Emerging data suggest links between diabetes (DM), cardiovascular disease, depression, and dementia. Both depression and dementia may present with impaired memory (IM) and may complicate chronic disease management and the ability to live autonomously for the elderly and disabled. [bold]Methods:[/bold] De-identified, cross-sectional data were extracted from the 2004 Medicare Current Beneficiary Survey in which 15559 beneficiaries residing, either in the community or long-term care facilities, or their caregivers were queried via interviews. General questions provided data about demographic traits, functional status, and disease burden. Neuro-psych questions addressed memory, concentration, decision making, sadness, anhedonia, and diagnoses of dementia. IM was defined either as memory loss or a dementia diagnosis. Supplemental questions provided data about DM. Descriptive and analytic statistics were used to characterize the population. [bold]Results:[/bold] 14500 community dwelling beneficiaries were identified ([ge]65 yrs: 11015, eligible by disability and [ge]65 yrs: 1003, and eligible by disability and [lt]65 yrs: 2482). In these 3 populations, the combined prevalence for pre/borderline diabetes (PBD) and DM was 19%, 32%, and 21% whereas the prevalence of coronary heart disease (CHD) overall [vs with PBD-DM] was 27% [38%], 43% [52%], and 18% [37%] respectively. Prevalence of IM was 11%, 17%, and 26% respectively. A dementia diagnosis occurred most frequently in beneficiaries eligible by age (33% of IM) and eligible by age + disability (23% of IM). Concomitant depression was common. Among age eligible beneficiaries, the proportion of patients with CHD was higher (37%) in those with IM than in those without (25%). Among age eligible beneficiaries, the proportion of patients with PBD-DM was higher (23%) in those with IM than in those without (19%). Those with PBD-DM and IM were younger (79 yrs) than those with IM alone (82 yrs) and older than those lacking IM with or without PBD-DM (76 and 77 yrs). The duration of PBD-DM was 6 years longer. A greater proportion of those with PBD-DM and IM had CHD (41%) than those with IM alone (37%) and those lacking IM with or without PBD-DM (36% and 22%). [bold]Conclusion: [/bold]The burden of DM and CHD is significant in all Medicare populations, but the interaction with IM differs by eligibility cohort. Further analysis will attempt to better characterize the interaction of these co-morbid conditions and assess their impact on functional status.[br][br]Nothing to Disclose: EAK, GSA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1594 180 1231 SUN-174 PO10-01 Sunday 1026 2012


1025 ENDO12L_SUN-175 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Change in Autonomic Regulation with Pioglitazone Monotherapy Is Related to Change in BMI Instead of Insulin Resistance among Asian Indian T2DM Patients Poonam Punjabi, Prashant Mathur, Deepak Kumar Gupta, Jyoti Thanvi, Sandeep Kumar Mathur SMS Medical College, Jaipur, India; SMS Medical College, Jaipur, India; Jaipur College of Pharmacy, Jaipur, India; Central University of Rajasthan, Kishangarh, India; SMS Medical College, Jaipur, India [bold]Aim:[/bold][br]To study effect of pioglitazone monotherapy on autonomic regulation, body weight and insulin resistance in treatment naive Asian Indian T2DM patients and relationship between changes in these parameters.[br][bold]Subjects and Methods: [/bold][br]Thirty-four newly diagnosed type-2 diabetes mellitus patients (47.21 [plusmn] 9.19 yrs M: F ratio 26:8) were treated with Pioglitazone 30 mg PO once daily for at least 6 months. Autonomic regulation was assessed by measurement of heart rate variability (HRV) by impedance plethysmograph recording of pulse wave in distal superficial arteries. Frequency domain analysis of HRV was carried out. Frequency domain parameters (Total Power, LF power, HF Power, LF (nu), HF (nu), LF/HF Ratio) were determined. Insulin Resistance was assessed by HOMA-R. Relationship between changes in parameters of HRV, insulin resistance and BMI at the beginning of treatment and after six months of therapy was studied. Student paired t test was used for comparison of baseline and follow-up parameters. Pearson correlation coefficient was used to find out relation between changes in parameters of HRV, HOMA- R and BMI.[br][bold]Results: [/bold][br]There was improvement in total power; decrease in LF (nu) and increase in HF (nu), but the differences were not statistically significant. However, there was positive correlation between change in BMI and change in HF (nu) (r= 0.34, p=0.04) and negative correlation between change in BMI and change in LF (nu) (r= -0.34, p=0.04). There was no association between change in HOMA-R and parameters of autonomic regulation.[br][bold]Conclusion:[/bold][br]In previous study we found that among Asian Indian T2DM patients parameters of autonomic regulation are related to body fat content instead of insulin resistance. The findings of present study, where intervention with a drug, which has bidirectional effect on these parameters i.e. increase in body weight but decrease in insulin resistance (contrary to in otherwise normal situation where increase in body weight is associated with increase in insulin resistance) further supports this hypothesis that insulin resistance is not the mediator of autonomic dysfunction among Asian Indian T2DM patients.[br][br]Nothing to Disclose: PP, PM, DKG, JT, SKM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1388 180 1232 SUN-175 PO10-01 Sunday 1027 2012


1026 ENDO12L_SUN-176 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Ischemic Peripheral Arterial Disease in Diabetic Patients Mehmet Calan, Firat Bayraktar, Dundar Ozalp Karabay, Tugba Gumus, Merve Yilmaz, Abdurrahman Comlekci Dokuz Eylul University Medical Faculty, Izmir, Turkey; Dokuz Eylul University Medical Faculty, Izmir, Turkey Purpose: We aimed to determine the prevalance of the ischemic peripheral arterial disease in type II diabetic patients[br]Material and method: The diabetic patients who were admitted to endocrinology outpatient clinic of DEU medical school hospital between May and August 2011 were randomly collected and included in the study. Their ages, genders, features and the diabetes durations were recorded. Weight, waist circumference, percent of body visceral fat distribution, and height were measured for each patient. Serum LDL cholesterol, HDL cholesterol, triglyceride, total cholesterol, creatinin, HgbA1C levels were recorded. ABI was evaluated for each lower extremity. The proportion of the highest systolic pressures of the lower extremity and the upper extremity was accepted as normal if ABI was [gt]0.9 and [lt]1.3, and abnormal if values were out of these limits.[br]Results: Study included 180 type II diabetes patients. Their average age was 59.4+-10.7 years, 53.9% was female and 46.1% was male, average BMI was 30.2+-5.3, mean body visceral fat distribution percent was 11.9+-4.2, mean HbA1C was 7.4 +-1.5.[br]PAD prevalance was 37.8% in the study group. According to gender, prevalance was 45.8% in male and 30.9% in female patients. When compared, PAD frequency was higher in male patients and this result was statistically significant (p[lt]0.05). The patients with PAD were questioned for peripheral arterial disease symptoms and only 10% defined claudication.[br]Conclusion:Type 2 diabetes mellitus is a major risk factor for peripheral arterial disease. Ankle brachial index is a simple and reliable method to screen peripheral arterial disease and to evaluate cardiovascular prognosis in the general population. Moreover, it has been reported that ABI is an independent marker for cardiovascular morbidity and mortality. ABI thresholds of less than 0.9 and more than 1.3 are highly suspicious for PAD and carries a high CV pathology risk in diabetic patients. Althouh PAD is very common in patients with diabetes, it remains under-recognised in this type of population. Diagnosis is often difficult when diabetes is associated with peripheral neuropathy because this could mask the pain. For this reason PAD may be asymptomatic until it reaches an advanced stage.[br]PAD prevalance was found to be 37.8% in diabetic patients. Only 10% of the patients with PAD had intermittent claudication symptom. Therefore we recommend a ABI evaluation, independent of claudication, in all type 2 diabetic patients.[br][br]Nothing to Disclose: MC, FB, DOK, TG, MY, AC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1494 180 1233 SUN-176 PO10-01 Sunday 1028 2012


1027 ENDO12L_SUN-177 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Evaluation of Biomarkers of Early Atherosclerosis in Adolescents with Type 1 Diabetes Mellitus Ozlem Engiz, Nazli Gonc, Ayfer Alikasifoglu, Alev Ozon, Nurgun Kandemir Hacettepe University, Ankara, Turkey [bold]Objective:[/bold] Type 1 diabetes mellitus (DM) is an important risk factor for the development of cardiovascular disease (CVD). We speculated that dislipidemia and atherosclerosis begins early in childhood in type 1 DM. The aim of this study is to evaluate the biomarkers of atherosclerosis and CVD risk factors in adolescents with type 1 DM considering glycemic control and diabetes duration.[br][bold]Study Design: [/bold]74 diabetic children, 22 children with metabolic syndrome and 38 healthy adolescents were included in the study. Children with type 1 DM were divided into three groups according to diabetes duration: group 1: [lt]5 years, group 2: 5-10 years, group 3: [gt]10 years. Diabetic children were also divided into two groups according to glycemic control: group 1: HbA1c[lt]8 and group 2: HbA1c[gt]8. BMI SDS, waist/hip circumference, systolic and diastolic blood pressure values of all participants were recorded. Serum fasting lipid profile, lipoprotein-a, apoprotein B, IL-6, TNF-[alpha], hs-CRP, ICAM-1, VCAM-1 and adiponectin levels of patients with type 1 diabetes were compared to those of adolescents with metabolic syndrome and healthy controls.[br][bold]Results:[/bold] Children with metabolic syndrome had higher biomarkers of atherosclerosis and lipid profiles compared to those of children with type 1 DM and healthy adolescents. Children with type 1 DM had significantly higher HDL cholesterol and lipoprotein-a levels than controls. Lipoprotein-a was higher in diabetics with HbA1c greater than 8 compared to ones with HbA1c less than 8(p[lt]0,05). Serum ICAM-1 and VCAM-1 levels were significantly increased in diabetic children compared with non-diabetic controls (p:[lt]0,001 and p:0,043 respectively). There were no significant differences regarding markers of inflammation, endothelial function and fibrinolytic activity between the groups when patients were grouped according to diabetes duration and glycemic control.[br][bold]Conclusions: [/bold]This study is the first study in literature that investigates the effects of glycemic control and diabetes duration on lipid profiles and biomarkers of early atherosclerosis in adolecents with type 1 DM. The adolescents with metabolic syndrome presented with the highest CVD risk when lipid profiles and atherosclerosis markers were evaluated. Endothelial dysfunction emerges in adolescents with type 1 DM but glycemic control and diabetes duration have no significant effect on markers of atherosclerosis. Lipoprotein a levels increase with poor glycemic control and long diabetes duration.[br][br]Nothing to Disclose: OE, NG, AA, AO, NK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 980 180 1234 SUN-177 PO10-01 Sunday 1029 2012


1028 ENDO12L_SUN-178 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) The Relationship between Sex Hormones, SHBG and Endothelial Function in Older Men: Data from the PIVUS Study Chiara Cattabiani, Gian Paolo Ceda, Fulvio Lauretani, Andrea Artoni, Rosalia Aloe, Giulia Schiavi, Marco Mantovani, Riccardo Volpi, Graziano Ceresini, Giuseppe Lippi, Tommy Cederholm, Lars Lind, Marcello Maggio University Hospital of Parma, Parma, Italy; University Hospital of Parma, Parma, Italy; University Hospital of Parma, Parma, Italy; Uppsala University Hospital, Uppsala, Sweden; Uppsala University Hospital, Uppsala, Sweden [bold][italic]Background. [/italic][/bold] Epidemiological studies have shown that low sex hormone binding globulin (SHBG) and testosterone (T) and high estradiol (E2) are independent predictors of metabolic syndrome and its components including hypertension in adult-older men (1).Preliminary data support a potential role of low SHBG in the risk of PAD in older men (2). However, despite the emerging link between SHBG, sex hormones and cardiovascular disease, the direct relationship with endothelium function has never been addressed. This is of importance since endothelium-dependent vasodilation assessed by the invasive forearm technique with acetylcholine given in the brachial artery (EDV) has been recently shown to be an independent risk factor of cardiovascular events in the older population (3).[br][bold][italic]Aim of the study.[/italic][/bold] To test the association between SHBG, sex hormones and endothelial function in older men.[br][bold][italic]Methods and Results. [/italic][/bold]Participants 70 years of age of the Prospective Study of the Vasculature in Uppsala Seniors (PIVUS) study (52% women), with complete data on EDV, and the brachial artery ultrasound technique with flow-mediated dilatation (FMD) were evaluated. From the entire sample of 1016 subjects, we used data of 430 men having information on SHBG, estradiol, T and endothelial function. SHBG, E2 and T were assessed by chemiluminescence (Beckman Coulter). The MDC for E2, T and SHBG were 73 pmol/L, 0.35 nmol/L and 2 nmol/L. The interassay coefficients of variation (CV) was [lt]20% for E2, the intra and inter CVs [lt]4 and [lt]7% for T, the interassay CV 5.3% for SHBG. Generalized linear models adjusted for multiple confounders including BMI, C Reactive Protein, fasting insulin, were used to test the relationship between sex hormones, SHBG and endothelial function. In a crude analysis we found a positive significant relationship between SHBG and EDV ([beta][plusmn] SE 4235.2 [plusmn] 908.5,p [lt].0001) but not with FMD. The relationship was maintained after adjustment for T ([beta][plusmn] SE 3734.6[plusmn] 1364.7, p=0.0065) and further adjustment for additional confounders including E2([beta][plusmn] SE 2898.9[plusmn] 1413. 1,p=0.04). No significant relationship was found between T, E2 and EDV or FMD.[br][bold][italic]Conclusions[/italic][/bold]. In older men SHBG, but not testosterone and estradiol, is positively and independently associated with endothelium-dependent vasodilation.[br][br]1. Brand JS, van der Tweel I, Grobbee DE, Emmelot-Vonk MH, van der Schouw YT. Testosterone, sex hormone-binding globulin and the metabolic syndrome: a systematic review and meta-analysis of observational studies. Int J Epidemiol. 2011;40(1):189-207. 2. Haring R, Travison TG, Bashin S, Vasan RS, Wallaschofski H, Davda MN, Coviello A, Murabito JM. Relationship between sex hormone concentrations, peripheral arterial disease, and change in ankle-brachial index: findings from the Framingham Heart Study. J Clin Endocrinol Metab 2011Dec;96(12):3724-32. 3. Lind L, Berglund L, Larsson A, Sundstr[ouml]m J.Endothelial function in resistance and conduit arteries and 5-year risk of cardiovascular disease.Circulation. 2011 Apr 12;123(14):1545-51.[br][br]Nothing to Disclose: CC, GPC, FL, AA, RA, GS, MM, RV, GC, GL, TC, LL, MM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 925 180 1235 SUN-178 PO10-01 Sunday 1030 2012


1029 ENDO12L_SUN-179 POSTER SESSION: Cardiometabolic Risk (1:30 PM-3:30 PM) Adiponectin and Resistin as Predictors of Subclinical Inflammation and Cardiometabolic Risk in Normoglycemic African Americans and Caucasians Sotonte Ebenibo, Alexander Boucher, Chimaroke Edeoga, Jim Wan, Samuel Dagogo-Jack University of Tennessee Health Science Center, Memphis, TN Background: Pro- and anti-inflammatory adipocytokines have been associated with cardiometabolic risk(CMR) and type 2 diabetes mellitus (T2DM). Because hyperglycemia induces a proinflammatory state, we studied the relationships among adipocytokines and CMR risk in normoglycemic subjects. We also assessed ethnic differences in the relationships.[br]Subjects and Methods: We studied 334 (153 white, 181 African American {AA}) enrollees (age 18-65 years) in our ongoing Pathobiology of Prediabetes in a Bi-racial Cohort study[sup]1[/sup]. After clinical assessment, fasting plasma specimens were obtained for measurement of glucose (FPG), lipids, adiponectin, resistin, and CRP, followed by a standard 75-gram OGTT. All subjects had normal OGTT. Metabolic syndrome (MetS) scores were recirded using NCEP cut-offs for waist, blood pressure, HDL and triglyceride levels. CRP served as a marker of inflammation.[br]Results: Compared to whites, AA subjects had lower mean ([plusmn]SD) FPG (91.0 [plusmn] 6.93 vs. 93.0 [plusmn] 6.29 mg/dl, P=0.0064) but higher BMI (women: 31.3 [plusmn] 6.94 vs. 29.1[plusmn] 7.24 kg/m[sup]2[/sup]; men: 30.6 [plusmn] 8.20 vs. 28.1 [plusmn] 6.17 kg/m[sup]2[/sup], P=0.0043). Adiponectin levels were higher in women than men (P [lt]0.0001) and lower in AA than whites (women: 8.75 [plusmn] 5.16 vs. 12.1 [plusmn] 5.67 ug/mL; men: 6.93 [plusmn] 3.85 vs. 7.36 [plusmn] 2.69 ug/mL, P [lt] 0.0001). In contrast, CRP levels were higher in women than men (P=0.0056) and higher in AA than whites (women: 5.82 [plusmn] 10.24 vs. 3.29 [plusmn] 4.82 mg/L; men: 3.44 [plusmn] 5.31 vs. 1.48 [plusmn] 3.53 mg/L, P=0.0034). Resistin levels differed by gender (women [gt] men, P= 0.0056) but not race. BMI was inversely related to adiponectin and positively related to resistin and CRP in AA and whites. MetS scores correlated inversely with adiponectin (AA: r= -0.30, P[lt] 0.0001; whites r= -0.27, P=.0007) and positively with resistin in whites (r= 0.21, P= 0.01) but not AA (r= 0.14, P=0.15). Disparities were noted in the relationships among adiponectin and CRP (whites: r= -0.16, P=0.05; AA: r= -0.11, P=0.15); adiponectin and resistin (whites: r=0.15, P=0.06; AA: r= 0.05, P= 0.52); and resistin and CRP (AA: r=0.14, P=0.06; whites: r=0.24, P=0.0033).[br]Conclusion: Among normoglycemic subjects, adiponectin predicts MetS in AA and whites, but has weaker interactions with CRP and resistin in African Americans. Resistin is a stronger predictor of cardiometabolic risk in whites than AA. These findings indicate ethnic disparities in the expression and putative functions of adipocytokine modulators of cardiometabolic/inflammatory risks.[br][br]1 Dagogo-Jack S, Edeoga C, Nyenwe E, Chapp-Jumbo E, Wan J. Pathobiology of Prediabetes in a Biracial Cohort (POP-ABC): Design and Methods. Ethn Dis 2011; 21:33.[br][br]Sources of Research Support: Grants from the NIH(R01 DK067269 and MO1 RR00211) and the American Diabetes Association.[br][br]Nothing to Disclose: SE, AB, CE, JW, SD-J 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1118 180 1236 SUN-179 PO10-01 Sunday 1031 2012


1030 ENDO12L_SUN-180 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) The Association of Estrogen Receptor-[beta] Gene with Salt-Sensitive Hypertension in Women but Not Men Chevon H Rariy, Bei Sun, Aditi R Saxena, Jonathan S Williams, Gordon H Williams, Ellen W Seely Brigham and Women[apos]s, Harvard Medical School, Boston, MA [bold]Background:[/bold]Genetic variations in the estrogen receptor have been associated with cardiovascular disease (CVD); most studies have focused on the estrogen receptor-[alpha] gene and not the [beta] gene (ESR2), which may be of more importance in women as ESR2 is the predominant estrogen receptor expressed in vascular smooth muscle cells. To further understand the role of ESR2 and mechanisms underlying hypertension, we investigated the relationship between single nucleotide polymorphisms (SNPs) of ESR2 and salt sensitivity of blood pressure. [bold]Methods:[/bold]In 365 white hypertensive men and women enrolled in HyperPATH, systolic blood pressure (SBP), mean arterial pressure (MAP) and renal blood flow (RBF) assessed by para-aminohippurate clearance, were measured on both low (10 mM Na/day) and high (200mM Na/day) salt diets. The changes in SBP(deltaSBP), MAP(deltaMAP) and RBF(deltaRBF) were calculated in response to sodium load. Gender-specific multivariate regression analysis was used to test the associations between SNP rs10144225 and blood pressure and RBF changes after accounting for age, body mass index, study site, and sibling relatedness. [bold]Results:[/bold]There was no association observed in men; however in hypertensive women, those who were homozygous for major allele A of rs1014425 as compared to minor allele G carriers had lower deltaSBP (15.95[plusmn]1.97 vs 24.20[plusmn]3.50mmHg, p=0.02) and deltaMAP (7.61[plusmn]0.54 vs 14.21[plusmn]0.93mmHg, p[lt]0.0001), but greater deltaRBF with borderline significance (28.52[plusmn]5.91 vs 3.41[plusmn]11.14cc/min, p=0.05). The associations were more prominent in premenopausal hypertensive women (age[lt]50y) with greater differences observed between rs10144225AA vs G carriers in deltaSBP (10.75[plusmn]3.7 vs 23.52[plusmn]5.78mmHg, p=0.02), deltaMAP (7.85[plusmn]1.2 vs 13.24[plusmn]2.1mmHg, p=0.05), and deltaRBF (31.17[plusmn]9.9 vs 6.39[plusmn]9.9cc/min, p[lt]0.0001). In postmenopausal hypertensive women (age[gt]50y), there was no association between rs10144225 and deltaSBP, deltaMAP, or deltaRBF. [bold]Conclusion:[/bold]There is a sex difference within ESR2 rs10144225, with an association present in white, hypertensive women, particularly in premenopausal women, but not men with Na+ sensitive changes in blood pressure and RBF. These findings suggest a role of ESR2 in the gender-specific development of hypertension, specifically in premenopausal women, possibly via defective modulation of renal perfusion when Na+ intake is changed.[br][br]Nothing to Disclose: CHR, BS, ARS, JSW, GHW, EWS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2319 181 1237 SUN-180 PO12-01 Sunday 1032 2012


1031 ENDO12L_SUN-181 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Clinical Characteristics of Patients Diagnosed with MEN 2A RET Mutation Carriers C634Y/Y791F Flavia Coutinho, Rodrigo Toledo, Tomoko Sekiya, Delmar Lourenco, Jr, Sergio Toledo University of S[atilde]o Paulo School of Medicine, S[atilde]o Paulo, Brazil Introduction/Objective: The majority of cases with multiple endocrine neoplasia type 2 (MEN2) has a single germline mutation in the RET gene. However, few cases ([sim] 16) with double mutations/polymorphisms in the RET gene described in the literature, usually associated with atypical phenotypes. Considering this the aim of this study was to characterize the clinical features of patients coming from five unrelated families, was identified double RET germline mutation (C634Y/Y791F).[br]Patients: Five index cases were recently discovered harboring the double RET germline mutation C634Y/Y791F. In these families there is a report of 208 potentially at-risk relatives (50%) to be carriers of this mutation. Among these 208 relatives, 72 patients (34.6%) agreed to participate in genetic screening.[br]Methods: We performed gene sequencing, covering all 20 exons of the RET gene. We investigated potential polymorphisms and mutations in the RET gene. MEN2-related tumors (medullary thyroid carcinoma, MTC, pheochromocytoma, PHEO, primary hyperparathyroidism, HPT) have been investigated and considered parameters such as: signs/symptoms, size, penetrance and aggressiveness of tumors, percentage of biochemical cure or recurrence of CMT.[br]Results: Of 72 individuals, 28 (38.9%) documented as carriers of RET double mutation C634Y/Y791F. It was observed that: a) Sixteen of 26 patients with MEN 2A (61.5%) had clinical diagnosis of FEO. The mean age at diagnosis was 43.8 [plusmn] 13.99 years, ranging from 27-66 years. Four of the five index cases had PHEO (80%) with aggressive features, such as tumors with size[gt] 5.0 cm, and a relatively early age at onset ([lt]35 years). The frequency of PHEO in adult relatives (from the third decade) mutation was also high (73%); Most cases presented PHEO bilateral (13/16; 81.25%). b) Twenty-four of the RET mutation carriers (24/27, 88.9%) had clinical diagnosis of CMT. The mean age at diagnosis of MTC was 37.8 [plusmn] 16.84 years, ranging from 7-66 years. Twelve of the 23 operated patients (52%) are so far considered biochemically [quot]cured.[quot] c) Among the 26 cases evaluated for the diagnosis of HPT, two of them (7.7%) were diagnosed with primary hyperparathyroidism.[br]Conclusion: We conclude that patients with this new double mutation tended to have more aggressive PHEO (early and well-endowed dimensions) and more frequently with bilateral PHEO, while CMT and HPT in these cases showed the usual carriers of mutation in codon 634.[br][br]Sources of Research Support: CNPq and FAPESP research grants.[br][br]Nothing to Disclose: FC, RT, TS, DL, ST 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2023 181 1238 SUN-181 PO12-01 Sunday 1033 2012


1032 ENDO12L_SUN-182 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Polymorphic Variation in the 5[prime] Regulatory Region of the [italic]CYP17A1[/italic] Gene Affects Transcription Efficiency Louise Diver, Samantha Alvarez-Madrazo, Frances McManus, Scott MacKenzie, John Connell, Eleanor Davies University of Glasgow, Glasgow, UK; University of Dundee, Dundee, UK [bold]Introduction:[/bold] Recent genome-wide association studies imply that the [italic]CYP17A1 [/italic]locus has a role in the regulation of blood pressure in man. This gene is important in steroidogenesis, regulating both glucocorticoid and androgen synthesis by catalysing 17[alpha]-hydroxylation and 17,20 lyase activity.[br][bold]Methods and Results:[/bold] We sequenced the entire [italic]CYP17A1 [/italic]locus in 60 normotensive Caucasian volunteers, establishing a pattern of linkage disequilibrium. Seven polymorphisms were identified in the promoter region and were assessed for putative transcription factor binding sites using the Transfac[reg] database. A polymorphism at position -362 (rs2486758), with allele frequencies T=0.76 and C=0.24 in our normotensive cohort, was further studied [italic]in vitro[/italic]. Differential transcriptional activity was assessed using reporter gene assays. Identical reporter constructs containing 2.9kb of the [italic]CYP17A1[/italic] promoter, varying only at the polymorphism site of interest, combined to firefly luciferase were co-transfected into H295R cells together with firefly renilla. A ratio of luciferase and renilla activity was used as an index of transcription in quadruplicate assays. Under basal conditions, the C allele (0.027 [plusmn] 0.003 RLU) showed greater activity then the T allele (0.017 [plusmn] 0.002 RLU, p=0.01). Under stimulated conditions (1mM Bu2cAMP for 24 hours), the C allele (0.057 [plusmn] 0.005 RLU) also showed higher activity than the T allele (0.033 [plusmn] 0.006 RLU, p=0.007). Experiments were replicated at least twice.[br][bold]Conclusions: [/bold]We have successfully demonstrated a detailed pattern of linkage disequilibrium across the [italic]CYP17A1[/italic] locus. [italic]In vitro[/italic] studies show that the C allele at -362 of the promoter region of [italic]CYP17A1 [/italic]is associated with a significantly greater transcriptional activity than the T allele, providing strong evidence that this locus may have functional significance. Further studies will examine the frequency of this polymorphism in a hypertensive population.[br][br]Nothing to Disclose: LD, SA-M, FM, SM, JC, ED 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1595 181 1239 SUN-182 PO12-01 Sunday 1034 2012


1033 ENDO12L_SUN-183 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Polymorphisms in the Glucocorticoid Receptor Family in Diverse Genetic Populations Shannon D Whirledge, Lisa Murphy, Stavros Garantziotis, John A Cidlowski National Institute of Environment Sciences, Reseach Triangle Park, NC; SRA International Inc, Durham, NC; National Institute of Environmental Sciences, Research Triangle Park, NC The diverse effects of glucocorticoids are mediated by the glucocorticoid receptor (GR). A large number of polymorphisms in the GR gene have been identified and linked to altered sensitivity of the receptor to glucocorticoids. An ATTTA-to-GTTTA (A3669G) polymorphism reported in the b isoform of GR, which acts as a dominant negative inhibitor of the active GR-a isoform, is associated with increased expression and stabilization of the b variant. Carriers with this polymorphism display diminished transrepressional activity of GR, important for mediating the immune system. Persons carrying the 9b haplotype appear to have a subtle but chronic pro-inflammatory status, leading to an increased risk of autoimmune diseases and risk for cardiovascular disease. An additional polymorphism has been identified in the b isoform (G3134T), however, no clinical studies have investigated whether this polymorphism is linked to any clinical features. Using the Environmental Polymorphism Registry (EPR), a racially diverse DNA repository of more than 15,000 individuals, we identified subjects with the well characterized A3669G polymorphism, as well as, the novel G3134T polymorphism. From the EPR, 4165 subjects were genotyped for the A3669G polymorphism. Carriers (C) represent 18.99% (791) of those screened and subjects homozygous (HC) for this polymorphism make up 1.6% (67) of the sample population. Those carrying at least one copy of this polymorphism, as well as non-carriers (NC), were invited to return for further clinical characterization. The primary endpoint for this study is the change in serum cortisol levels following a modified dexamethasone suppression test. Additional population demographics will be recorded, including: race, ethnicity, sex, age, body temperature, BMI, blood pressure, and respiratory rate, as wells as a health history report. Currently, 67 of 102 subjects have been enrolled to detect an effect size of 75 nmol/L at 80% power. These subjects have also been screened for the G3134T polymorphism. Interestingly, of those NC for A3669G, 57.1 % are C for G3134T and 28.6% are HC for this mutation. The two polymorphisms may in fact be linked. Those who are C for A3669G have an increased incidence of G3134T, 88% as C and 40% HC. This is further amplified in HC of A3669G, 100% are C of G3134T and 83% are HC for the G3134T polymorphism. Characterization of this polymorphism may provide evidence for how genetic variations of the glucocorticoid receptor function.[br][br]Nothing to Disclose: SDW, LM, SG, JAC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1615 181 1240 SUN-183 PO12-01 Sunday 1035 2012


1034 ENDO12L_SUN-184 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Congenital Hyperinsulinism Caused by a Novel Mutation in the ABCC8 (SUR1) Gene Sadana Balachandar, Saroj Nimkarn, Zoltan Antal Weill Cornell Medical College, New York, NY Background:[br]Congenital hyperinsulinism (CH) is the most common form of persistent hypoglycemia in neonates and children. Alterations in 7 genes are associated with CH, but KCNJ11 and ABCC8 mutations account for the majority of cases. The KCNJ11 and ABCC8 encode two important subunits of the membrane KATP channel, Kir6.2 and SUR1. KATP channel malfunction leads to an inappropriate release of stored insulin. Typically, patients with these mutations are large for gestational age and present with severe hypoglycemia during the first days of life, as well as apneic episodes, hypotonia and poor feeding.[br]Objective:[br]We report the case of a 13 month old girl with a novel sequence variant of the ABCC8 gene.[br]Design/Methods:[br]Clinical evaluation was performed in the inpatient setting, with subsequent outpatient follow-up.[br]Results:[br]We report the case of 13 month old female with recurrent hypoglycemic seizures. She was born full-term and appropriate for gestational age. One hypoglycemic event occurred in the neonatal period, but resolved with feeding alone. Normal growth and development ensued in the first 10 months of life. At 11 months of age, she developed a generalized tonic clonic seizure associated with hypoglycemia. Neurological evaluation did not reveal an underlying cause of her seizure. At 13 months of age, she came to our institution following another hypoglycemic seizure. During her hospital course, she developed hypoglycemia within a few hours post meals, and responded to glucagon with a 68 mg/dL rise in serum glucose. Her clinical diagnosis was established based on the absence of ketosis, elevated insulin levels and normal pituitary hormones in the face of hypoglycemia. Analysis of several genes causing CH revealed a novel sequence variant in the ABCC8 gene. It was a paternally inherited C to T variant in nucleotide position 822+20. She had an excellent outcome after diazoxide treatment, 10 mg/kg/day, without further episodes of hypoglycemic seizures.[br]Conclusions:[br]This is the first case report demonstrating CH in a region of the ABCC8 gene not typically associated with CH. Our case differs from the typical CH child with an ABCC8 gene mutation in that she was appropriate for gestational age with severe symptoms presenting after 11 months of age, rather than large for gestational age with severe hypoglycemia persisting in the neonatal period. Our case demonstrates a novel mutation in the ABCC8 gene associated with an atypical presentation of CH.[br][br]Nothing to Disclose: SB, SN, ZA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 215 181 1241 SUN-184 PO12-01 Sunday 1036 2012


1035 ENDO12L_SUN-185 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Variants in Type 2 Diabetes Susceptibility Genes and Other Loci Affect Risk of Cystic Fibrosis-Related Diabetes (CFRD): The International CF Gene Modifier Consortium Scott M Blackman, Clayton W Commander, Christopher Watson, Lisa J Strug, Johanna M Rommens, Rhonda G Pace, Sarah A Norris, Peter R Durie, Mitchell L Drumm, Michael R Knowles, Garry R Cutting Johns Hopkins University School of Medicine, Baltimore, MD; Johns Hopkins University School of Medicine, Baltimore, MD; University of North Carolina, School of Medicine, Chapel Hill, NC; University of Toronto and Hospital for Sick Children, Toronto, Canada; Case Western Reserve University, Cleveland, OH [bold]Background:[/bold] Diabetes is a common complication in adolescents and adults with cystic fibrosis (CF) in which patients generally have normal insulin sensitivity but deficient insulin production. The cause of CF-related diabetes (CFRD) is unknown, but we have previously shown that CFRD shares genetic susceptibility with type 2 diabetes (T2D): CFRD clusters in families with type 2 diabetes (T2D), and three T2D susceptibility genes ([italic]TCF7L2[/italic], [italic]CDKAL1[/italic], [italic]CDKN2A/B[/italic]) are also genetic modifiers of CFRD (ref 1 and unpublished).[br][bold]Objective:[/bold] We are testing single nucleotide polymorphisms (SNPs) genome-wide for association with CFRD.[br][bold]Design/Methods:[/bold] Age of onset of CFRD was treated as a survival trait (event = CFRD diagnosis; censor = last negative test). Genotypes were obtained for 3,899 patients in the CF Twin and Sibling study (TSS), Canadian Consortium for CF Genetic Studies (CGS), and CF Gene Modifiers Study (GMS), using the Illumina 610-Quad array with 564,637 SNPs analyzed. Study-specific P-values were combined by meta-analysis as we did for CF lung function (ref 2). An independent replication sample of 704 individuals was genotyped using TaqMan.[br][bold]Results:[/bold] Genome-wide significant evidence for association with CFRD was found for SNPs in [italic]SLC26A9[/italic] (HR=1.37 [95% CI: 1.23-1.54]; P=3.9[times]10[sup]-08[/sup]; n=3060). Effect size (hazard ratio) was similar in each study and in patients homozygous for F508del mutations in [italic]CFTR[/italic]. There was no evidence for allelic heterogeneity. The same alleles associated with CFRD in the replication sample (HR=1.48 [1.12-1.95]; P=0.006; n=704). [italic]SLC26A9[/italic] encodes a chloride/bicarbonate transporter which is known to interact functionally with CFTR (ref 3). Together, 4 identified CFRD modifiers more than triple the CFRD risk in this population.[br]Using regional association we follow-up on our genome-wide linkage study which provided evidence for linkage at chromosome 2q35-2q36.1 (LOD = 3.8, approximately equivalent to study-wide P=0.035) and chromosome 12p12 (LOD = 2.24). Region-wide significant evidence for association was found for a noncoding variant at 12p12 (P=1.5[times]10[sup]-05[/sup]; 1205 SNPs tested). Ongoing studies include sequencing to identify candidate causal variants.[br][bold]Conclusions:[/bold] Common genetic variants can substantially impact the risk of CFRD and may be of utility for identifying high-risk patients. Type 2 diabetes genes play a role in CFRD, suggesting that some disease pathways may act in both forms of diabetes. Gene variants may also act through CFTR-specific pathways.[br][br]1. Blackman SM et al, Diabetologia 2009 52:1858-65. 2. Wright FA et al, Nat Genet. 2011 43:539-46. 3. Strug LJ et al., Nat Genet 2012 In press.[br][br]Sources of Research Support: LWPES, NIH DK083551, CFF (S.M.B.); NIH DK044003, HL068927, CFF (G.R.C.), HL068890, DK066368, RR000046, CFF (M.R.K.), CFC (L.J.S.).[br][br]Nothing to Disclose: SMB, CWC, CW, LJS, JMR, RGP, SAN, PRD, MLD, MRK, GRC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2131 181 1242 SUN-185 PO12-01 Sunday 1037 2012


1036 ENDO12L_SUN-186 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Association of the [italic]CD226[/italic] Variant (rs763361) with Autoantibodies Related to Multiple Autoimmune Diseases in a Cohort of Brazilian Type 1 Diabetes Aritania Sousa Santos, Teresa Cristina Mattana, Vinicius Silva Costa, Debora Teixeira Mainardi-Novo, Rosa Tsunecniro Fukui, Rosa Ferreira Santos, Maria Elizabeth Rossi Silva Hospital das Cl[iacute]nicas, S[atilde]o Paulo, Brazil; Faculdade de Medicina da Universidade Federal de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Genetic basis of autoantibody production outside of the HLA region is not totally understood and deserves more knowledge. Recently the nonsynonymous polymorphism rs763361 (A/G), Gly307Ser, of the [italic]CD226[/italic] gene has been identified as a genetic risk factor for autoimmunity. However, its role in susceptibility to several autoimmune diseases in patients with type 1 diabetes (T1D) has not been analyzed.[br]Objective: To evaluate the association of rs763361 variant of [italic]CD226[/italic] with autoantibody positivity for multiple autoimmune diseases in a Brazilian cohort of TD1 patients.[br]Patients and methods: 535 T1D patients (mean age 22.2[plusmn]14.9 years, 55.3% female, and 81% of European ancestry) were genotyped for the rs763361 variant of [italic]CD226 [/italic]and compared with a group of 591 health controls (mean age 28[plusmn]11.5 years, 38.4% female, and 61.75% of European ancestry). The presence of autoantibodies was also evaluated. Chi-square and Fisher[apos]s test (contingency tables 2x2) were used for statistical comparisons.[br]Results: The rs763361 genotype distribution was in Hardy Weinberg equilibrium in both groups (P=0.0775 and P=0.5186). The AA genotype prevailed in T1D patients (36.7% x 25.7%-OR=1.46; CI= 1.132-1.894; P=0.0044) whereas the AA+AG genotypes increased the frequency of anti-glutamate decarboxilase (GADA) antibody in a subgroup of T1D patients with the disease for less than 2 years. The presence of A allele did not predispose to anti-islet antigen-2 (IA-2A) antibody positivity. Considering other autoimmune diseases, both the AA and AA+AG genotypes did not favor the presence of the following autoantibodies: anti-thyreoperoxidase (TPO), anti-thyroglobulin (Tg), thyrotropin receptor (TRAb), antineutrophils cytoplasmic antibodies (ANCA), anti-gastric parietal cell, anti-smooth muscle, anti-endomysium, anti-nuclear and anti-21 hydroxylase. Interestingly, the A allele and the AA genotype protected to rheumatoid factor positivity (6.7% x 10.8%; OR=0.592, CI=0.373-0.94, P=0.033 and 4.4% x 11.3%; OR= 0.361; CI=0.148-0.882; P=0.02 respectively) among both T1D patients and controls.[br]Conclusion: The rs763361 variant in [italic]CD226 [/italic]increased the risk to type 1 diabetes and to anti-GADA antibody positivity, whereas decreased the frequency of rheumatoid factor. No association of this polymorphism with other organ-specific autoantibodies was found in the T1D population.[br][br](1)Todd JA, et al., Nature Genetics 2007;39:857. (2)Maiti AK et al., Rheumatology 2010;49:1239.[br][br]Sources of Research Support: FAPESP.[br][br]Nothing to Disclose: ASS, TCM, VSC, DTM-N, RTF, RFS, MERS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1146 181 1243 SUN-186 PO12-01 Sunday 1038 2012


1037 ENDO12L_SUN-187 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) The Genetic Basis of Maturity-Onset Diabetes of the Young (MODY) in Cyprus Christos Shammas, Vassos Neocleous, Nicos Skordis, Leonidas A Phylactou The Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus; Makarios III Hospital, Nicosia, Cyprus Maturity-onset diabetes of the young (MODY) is an inherited non-insulin-dependent form of diabetes. It is autosomal dominant and disturbs important processes during beta cell development, function and regulation. The two most common forms of MODY are type 2 and 3. MODY2 is caused by mutations in the GCK gene and impairs glucokinase activity that results in stable, mild hyperglycaemia that rarely requires treatment. MODY3 is caused by mutations in the HNF1A gene that can develop to a progressive insulin secretory defect that is sensitive to sulphonylureas. Mutation analysis of 20 Cypriot families that presented symptoms of MODY are described.[br]Genetic testing revealed four patients with mutations in the GCK gene of which two are novel. The two novel mutations are p.E440Stop and p.R447P while common mutations p.G261R and p.R43H have also been identified. Novel GCK mutations p.E440Stop and p.R447P display a mild fasting hyperglycemia phenotype with only mild glucose intolerance. Nonsense mutation p.E440stop terminates the cytoplasmic enzymatic activity since it is responsible for the loss of the C-terminal end of the GCK peptide that includes vital residues required for the release of [alpha]13 helix during glucose binding. p.R447P is located in the loop between [beta]13 and [alpha]13 domains of the Gucokinase enzyme that has an important role during the super-open to the closed active conformation of the enzyme; p.R447P mutation favors the transition to the closed conformation.[br]Eight patients with HNF1A variants p.I27L, p.A98V, p.G288G and p.S487N have been found. These variants have demonstrated to decrease transcriptional activity upon genes involved in glucose metabolism. In our cohort of patients identified with HNF1A variants, p.I27L which is linked with insulin resistance was the most frequent (38.5%), followed by p.S487N (30.8%), p.G288G (23.6%) and p.A98V (7.6%).[br]In conclusion, the present study can be used to identify the essential components required for glucose homeostasis and determine the allosteric site of glucokinase that can be potentially targeted by drugs to treat type 2 diabetes. In addition this findings will be used towards the effective diagnosis of MODY, improve genetic counseling and used as a potential therapeutic platform in the future for the affected patients in Cyprus.[br][br]Nothing to Disclose: CS, VN, NS, LAP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 393 181 1244 SUN-187 PO12-01 Sunday 1039 2012


1038 ENDO12L_SUN-188 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Effect of Calpain-10 and Adiponectin Gene Polymorphism on Insulin Secretion and Insulin Resistance of Type 2 Diabetes Mellitus Youngmi Lee, Min Kyung Kim, Jiyoon Ha, Eun Jin Kwon, Jiwoon Kim, Tae Woong No, Shinae Kang, Jong Suk Park, Chul Woo Ahn, Kyung Rae Kim Yonsei University Colleage of Medicine, Seoul, Republic of Korea It is thought that genetic variations in the Calpain-10 and adiponectin gene effect insulin secretion in the patients with type 2 diabetes mellitus. Recently, several single nucleotide polymorphisms (SNPs) in the adiponectin and Calpain-10 gene have been reported to be associated with type 2 diabetes and components of the insulin resistance and secretion.[br]The aim of this study was to determine whether there is an association between specific CAPN10, adiponectin gene and T2DM in the Korean population. Futhermore evaluate association of genetic variations in the Calpain-10, adiponectin gene and metabolic syndrome.[br]Overall, 249 Korean patients with T2DM and 131 non-diabetic controls with no family history of diabetes were enrolled in this study.[br]G276T and T45G frequencies of the adiponectin gene was examined in 249 unrelated type 2 diabetic and 131 non-diabetic control Korean subjects. All the subjects were genotyped according to CAPN10 SNP-43 and -63. The clinical characteristics and genetic levels of the subjects were compared within these genotypes.[br]Among 249 to and 131 control evaluated genetic polymorphism. Variation in calpain 10, SNP-63 T/T showed higher association in type 2 diabetes. The results of this study suggest that single SNP-63 increases the susceptibility to type 2 diabetes. But calpain 10, SNP -43, adiponectin T45G and intron G276T genetic frequency is not associated.[br]Variation in calpain 10, SNP-63 plays a role in the susceptibility to type 2 diabetes individually. SNP-43 and Adiponectine SNP-45, -276 are not associated with glucose tolerance.[br][br]Nothing to Disclose: YL, MKK, JH, EJK, JK, TWN, SK, JSP, CWA, KRK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1356 181 1245 SUN-188 PO12-01 Sunday 1040 2012


1039 ENDO12L_SUN-189 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Protein Biomarkers in Prognosis of Molecular Pathogenesis in First-Degree Relatives of Type 2 Diabetics Nabila Roohi University of the Punjab, Lahore, Pakistan [bold]Background and aims: [/bold]First degree relatives (FDR) of type 2 diabetics are at a higher risk of developing diabetes. The risk is continually increasing with increasing prevalence of type 2 diabetes mellitus (T2DM) worldwide. Identifying the predictors associated with susceptibility to T2DM, therefore, becomes increasingly important.[br][bold]Subjects and Methods:[/bold] Participants were age and sex matched type 2 diabetics (n=20); their first degree relatives (positive controls, PC) having parents with conjugal T2DM, and including both normal (NGT, n=15) and impaired glucose tolerance (IGT, n=15) subjects; negative controls (NC, n=10) with no clinical and/or family history of DM. Serum proteins, resolved by SDS-PAGE, quantified, electroblotted, pertinent fractions excised out, electroeluted, run on isoelectric focusing gels and identified using Swiss 2DPAGE database. The correlation of T2DM with resolving serum protein patterns was assessed to identify the specific protein/s linked with T2DM and the patterns were compared with those of FDRs to find the possibility of early prediction of diabetes in asymptomatic subjects.[br][bold]Results: [/bold]C-reactive protein (CRP), Apolipoprotein (Apo) A-IV and ApoE were significantly over expressed (p[lt]0.05) in sera of type 2 diabetics as compared to NC. Similar trends of elevated expressions (p[lt]0.05), of these proteins, were also observed in all IGT and most NGT subjects of positive controls category indicating their predisposition to the future development of T2DM.[br][bold]Conclusions: [/bold]Enhanced expressions of CRP, ApoA-IV and ApoE, in type 2 diabetics indicate that these proteins may be regarded as independent risk determinants of molecular pathogenesis in T2DM. Persistence of similar trends in most of the PC strongly suggests that these proteins could be employed as biomarkers in early prediction of the disease in FDRs. The study further suggests that IGT relatives of type 2 diabetics are the highest risk category amongst FDRs. Regular screening of FDRs, at population level, is recommended in order to prevent or delay the manifestation of diabetes and its complications in these subjects.[br][br]Nothing to Disclose: NR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1019 181 1246 SUN-189 PO12-01 Sunday 1041 2012


1040 ENDO12L_SUN-190 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Functional Analysis of Genetic Variants and Vitamin D Status in Immune Cells by RT-PCR Yasmin Moran-Auth, Marissa Penna-Martinez, Elisabeth Ramos-Lopez, Klaus Badenhoop University Hospital Frankfurt am Main, Frankurt am Main, Germany [bold]Aim: [/bold]Vitamin D insufficiency has been linked to Type 1 Diabetes and studies related about significant correlations with polymorphisms of vitamin D ([italic]CYP2R1[/italic], [italic]CYP27B1[/italic]) and cholesterol biosynthesis ([italic]DHCR7[/italic]) genes. To investigate possible associations, gene expression in immune cells, vitamin D status and genetic variants were analyzed in healthy controls (HC). [bold]Methods: [/bold]From 23 HC monocytes (Mo), T-helper cells (Th) and natural killer cells (NK) were isolated and after cell lysis extracted RNA was reverse transcribed into cDNA. Furthermore gene expression of DHCR7, VDR, CYP2R1, CYP27B1 and the endogenous control GAPDH were measured by Taqman assay. Finally,[italic] DHCR7 [/italic](rs-12785878[italic]), CYP2R1 [/italic](rs-10741657), [italic]CYP27B1 [/italic](rs-10877012), [italic]VDR Fok [/italic](rs-1073580) polymorphisms were genotyped and 25-hydroxyvitamin D[sub]3[/sub] [25(OH)D[sub]3[/sub]] plasma levels were measured using radioimmunoassay. Statistical analyses were performed using the 2[sup]-delta-delta-Ct[/sup] values by Kruskall-Wallis-test. [bold]Results: [/bold]All studied immune cells showed a significantly different gene expression of DHCR7 (NK=100, Mo=10, Th=116x10[sup]3[/sup]; p=0.001), CYP2R1 (NK=238, Mo=20, Th=162x10[sup]3[/sup]; p=0.001) and CYP27B1 (NK=15, Mo=6, Th=72x10[sup]4[/sup]; p=0.001). By dividing the HC into 25(OH)D[sub]3[/sub] deficiency[sub] [/sub]([lt]20ng/ml) and 25(OH)D[sub]3[/sub] sufficiency ([gt]20ng/ml), significant down regulation of VDR expression in NK and Th cells from HC with vitamin D deficiency compared to those with vitamin D sufficiency was observed (p=0.019 and p=0.037 respectively). No associations between the gene expression levels and the investigated polymorphisms in all immune cells were detected. However, the CYP27B1 expression was significant higher in NK cells from HC with vitamin D deficiency and the [ldquo]CC[rdquo] CYP27B1 genotype than those with the [ldquo]AC[rdquo] genotype (p=0.03). [bold]Discussion:[/bold] Vitamin D deficiency or its deficiency in combination with the CYP27B1 polymorphism seems to interact with cellular immune effects like VDR or CYP27B1 expression illustrating the need for an individualized therapy.[br][br]Sources of Research Support: This study was supported by FP7-NAIMIT, grant agreement No. 241447 is kindly acknowledged.[br][br]Nothing to Disclose: YM-A, MP-M, ER-L, KB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 812 181 1247 SUN-190 PO12-01 Sunday 1042 2012


1041 ENDO12L_SUN-191 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Anxiety Linked to a T2D Gene Claudia Gragnoli Pennsylvania State College of Medicine, Hershey, PA; Temple University[apos]s College of Science [amp] Technology, Philadelphia, PA; Bios Biotech Multi-Diagnostic Health Center, Rome, Italy Background: The chromosome 12q24 locus is linked to bipolar disorder, depression, anxiety, and type 2 diabetes (T2D). Proteasome Modulator 9 (PSMD9) lies in the 12q24 locus and is linked MODY3, T2D, T2D-micro- and macro-vascular complications, hypercholesterolemia, depression and insomnia. Interestingly, PSMD9 is implicated in the response to anti-depressant treatment. PSMD9 is ubiquitously highly concentrated in eukaryotic cells as part of the 26S proteasome complex, contributing to intracellular proteins degradation into antigenic peptides in the immune response to antigen presentation by MHC class I cells thereby potentially playing a causative role in inflammation. Further, PSMD9 mediates the transcription of the ligand-dependent retinoid-target genes. Aims: Our goal was to determine whether PSMD9 is linked to anxiety in Italian T2D families. Methods: We phenotyped the Italian families[apos] members for presence and/or absence of anxiety using the diagnostic criteria of DSM-IV. The phenotype was described as unknown in all cases in which the diagnosis was unclear or data were not available. We tested in the 200 Italians families for evidence of linkage of the PSMD9 T2D risk single nucleotide polymorphisms (SNPs) IVS3+nt460A[gt]G, IVVS3+nt437T[gt]C and E197G A[gt]G with the anxiety phenotype. The non-parametric linkage analysis was performed by using the Merlin software. To rule out results due to random chance, 1000 replicates were executed. Results: The PSMD9 T2D risk SNPs studied are linked to anxiety in our Italian families. Conclusions: This is the first report of the PSMD9 T2D gene in linkage to anxiety. The implication of this finding underlines an innovative link of anxiety to T2D, as well as the potential PSMD9 pleiotropic effect on the pathogenesis of associated complex disorders.[br][br]Sources of Research Support: Penn State College of Medicine, Hershey, PA. Bios Biotech Multi-Diagnostic Health Center, Rome, Italy.[br][br]Nothing to Disclose: CG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1111 181 1248 SUN-191 PO12-01 Sunday 1043 2012


1042 ENDO12L_SUN-192 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Multivariate Data Analysis from the National Health and Nutrition Examination Survey (NHANES) 2001[ndash]2006 Shows That Second-Hand Smoke Is Associated with Both Obesity and Diabetes Mellitus Dulcie Kermah, Magda Shaheen, Deyu Pan, Theodore C Friedman Charles R Drew University of Medicine [amp] Science, Los Angeles, CA; Charles R Drew University of Medicine [amp] Science, Los Angeles, CA Studies have shown a positive association between cigarette smoking and rate of type 2 diabetes (DM), an association that is surprising as smokers are leaner than non-smokers and obesity is positively associated with DM. However, the relationship between second-hand smoke and DM is less established; especially missing is studies which verified second-hand smoke with cotinine levels. Our objective was to investigate the relationship between cotinine level verified second-hand smoke and both DM and obesity. We examined data from 6,391 adults from the NHANES, 2001-2006, a nationally representative sample of the United States population. Current smokers (25%) were defined as answering [ldquo]yes[rdquo] to the question (Q1), [ldquo]do you smoke cigarettes?[rdquo] and having a serum cotinine [gt] 3.0 ng/mL. Second-hand smokers (34%) were defined as answering [ldquo]no[rdquo] to Q1 and having a serum cotinine [gt] 0.05 ng/mL. Non-smokers (41%) were defined as answering [ldquo]no[rdquo] to Q1, and having a serum cotinine [lt] 0.05 ng/mL. The outcome variables were body mass index (BMI), fasting blood glucose (FBG), HOMA and HbA1c. Using a multivariate model adjusting for age, sex, race, alcohol and physical activity, second-hand smokers had higher HOMA ([beta]=0.08[plusmn]0.04; p=0.048), HbA1c ([beta]=0.01[plusmn]0.004; p=0.01), FBG ([beta]=0.02[plusmn]0.009; p=0.02) and BMI ([beta]=0.02[plusmn]0.009; p=0.02) relative to non-smokers. Current smokers had higher HbA1c ([beta]=0.01[plusmn]0.005; p=0.01) and lower BMI ([beta]=-0.03[plusmn]-0.01; p=0.01) relative to non-smokers. Adding BMI to the adjusted model, second-hand smokers and current smokers had higher HbA1c [([beta]=0.011[plusmn]0.005; p=0.02); ([beta]=0.018[plusmn]0.005; p=0.001) respectively] relative to non-smokers. We conclude that second-hand smoke is positively associated with obesity and DM and the association with DM was not through obesity. More studies are needed to show a causal relationship between second-hand smoke and DM and to understand the mechanisms involved in the association.[br][br]Nothing to Disclose: DK, MS, DP, TCF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1387 181 1249 SUN-192 PO12-01 Sunday 1044 2012


1043 ENDO12L_SUN-193 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Association of Obstructive Sleep Apnea and Glucose Metabolism in Nonobese Koreans Ji A Seo, Chai Ryoung Eun, Hyunjoo Cho, Ho Cheol Hong, Hae Yoon Choi, Yoon Jung Kim, Joo Hyung Kim, Sae Jeong Yang, Hye Jin Yoo, Hee Young Kim, Sin Gon Kim, Kyung Mook Choi, Sei Hyun Baik, Dong Seop Choi, Chol Shin, Nan Hee Kim Korea University Medical School, Seoul, Republic of Korea; Korea University Medical School, Seoul, Republic of Korea [bold]Background[/bold]: To investigate whether the association between obstructive sleep apnea (OSA) and glucose metabolism is different according to the obesity status and these associations were consistent after adjusting for visceral obesity in Korean adults.[br][bold]Methods[/bold]: A total of 844 Koreans (561 men and 283 women) were enrolled from the Korean Genome Epidemiology Study, which is an ongoing population-based cohort study. All participants underwent polysomnography (PSG) with a computerized PSG device. OSA was assessed using the apnea-hypopnea index (AHI), defined as the average number of apneic and hypopneic events per hour of sleep. The participants were classified according to the presence of sleep apnea [AHI [lt] 5/h (no-OSA), [ge] 5/h (OSA)] and obesity [BMI [lt]25kg/m[sup]2 [/sup](non-obese), [ge]25kg/m[sup]2 [/sup](obese)]. Using 75g oral glucose tolerance test, the participants were categorized into normal glucose tolerance, impaired fasting glucose (IFG only), impaired glucose tolerance (IGT only), IFG plus IGT, and diabetes.[br][bold]Results[/bold]: In the non-obese group, the subjects with OSA had higher fasting plasma glucose (FPG), HbA1c and HOMA-IR levels than those without [FPG, 103.2[plusmn]27.2 vs. 96.6[plusmn]21.8 mg/dL, p=0.005; HOMA-IR (median, IQR), 1.9(1.4-2.6) vs. 1.7(1.3-2.1), p[lt]0.001; HbA1c, 5.5(5.2-5.8) vs. 5.0(5.4-5.9)%, p=0.001, OSA vs. no-OSA subjects, respectively]. However, in the obese group, only HOMA-IR levels were higher in OSA than no-OSA subjects [median(IQR), 2.7(1.9-3.3) vs. 2.2(1.7-3.0), p=0.009]. The prevalence of IFG plus IGT was higher in OSA subjects regardless of obesity [OSA vs. no-OSA, 20.5 vs. 7.0% in non-obese, p=0.002; 29.9 vs. 15.9 % in obese, p=0.047]. Interestingly, the impact of OSA on the prevalence of diabetes was evident only in non-obese subjects (OSA vs. no-OSA, 36.2 vs. 16.4% in non-obese, p[lt]0.001; 39.4 vs. 31.3%, p=0.1 in obese). This relationship was consistent in the multivariate regression analysis. The odd ratio (OR) for diabetes was significantly higher in those with OSA than no-OSA in the non-obese subjects [OR(95% CI), 1.9(1.1-3.4) vs. 0.9(0.6-1.5), non-obese vs. obese subjects, respectively) after adjusting for age, sex, visceral fat area, cardiovascular disease, medication for hypertension, smoking, alcohol drinking, and exercise.[br][bold]Conclusion[/bold]: The association between OSA and diabetes was significant only in non-obese Koreans even after adjusting for major risk factors for diabetes including visceral obesity.[br][br]Nothing to Disclose: JAS, CRE, HC, HCH, HYC, YJK, JHK, SJY, HJY, HYK, SGK, KMC, SHB, DC, CS , NHK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 303 181 1250 SUN-193 PO12-01 Sunday 1045 2012


1044 ENDO12L_SUN-194 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Gender-Specific Association of Vitamin D and Insulin Resistance among High-School Students Hyeon Chang Kim, Dong Phil Choi, Yumie Rhee Yonsei University College of Medicine, Seoul, Republic of Korea; Yonsei University College of Medicine, Seoul, Republic of Korea [bold]BACKGROUND[/bold]: Low serum levels of 25-hydroxyvitamin D (25-OHD) are associated with type 2 diabetes or insulin resistance in adults. However it is still unclear low-OHD level is associated with insulin resistance especially among healthy adolescents. Thus we investigated association between 25-OHD levels and homeostasis model assessment-insulin resistance (HOMA-IR) among apparently healthy adolescents.[br][bold]METHODS: [/bold]This is a cross-sectional analysis of baseline data collected for an adolescent cohort study. Health examinations and questionnaire survey were conducted for all first grade students (mean age 15.5 years) at a rural high school in Jang Seong, South Korea, from June 19 to 22, 2011. Of the 268 first -year students, 261 (136 male and 125 female) students were enrolled in the study. Overnight fasting blood samples were drawn to measure metabolic profile including fasting glucose, insulin, and serum 25-OHD. Spearman correlation analysis, multiple linear and logistic regression analyses were preformed to assess the association between 25-OHD and HOMA-IR. The study protocol was approved by the institutional review board. Informed consent was obtained from all participating students and their parents.[br][bold]RESULTS: [/bold]In male adolescents, 25-OHD level was significantly correlated with fasting insulin (correlation coefficient r=-0.256; p=0.003) and HOMA-IR (r=-0.263; p=0.002) but not with fasting glucose (r=-0.153; p=0.075). Even after adjustment for age, body mass index, smoking, alcohol intake, and physical activity, low serum 25-OHD level was significantly associated with higher insulin level (p=0.005) and HOMA-IR (p=0.004). When serum 25-OHD levels were categorized into quartiles, boys in the lowest quartile had 3.60 times higher odds ratio (95% confidence interval 1.03 to 12.63) for having increased insulin resistance, compared to those in the highest quartile. However, in female adolescents, 25-OHD level was not associated with glucose, insulin, or HOMA-IR.[br][bold]CONCLUSION: [/bold]Our findings partially support that low serum 25-OHD level is associated with insulin resistance among healthy adolescents. Gender difference in the association between 25-OHD and insulin resistance needs to be further investigated.[br][br]Sources of Research Support: This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (Grant No. 2010-0007860, 2011-0005131).[br][br]Nothing to Disclose: HCK, DC, YR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1183 181 1251 SUN-194 PO12-01 Sunday 1046 2012


1045 ENDO12L_SUN-195 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Epidemiologic Characteristics of Diabetes in Children Aged 0[ndash]14 Years in Busan and Gyeongnam Province, Korea (2001[ndash]2010) Jung Hyun Lee, Min Jung Kwak, Su Yung Kim, Hyun-Ji Kim, Sun Hee Lee, Chong Kun Cheon, Woo Yeong Chung, Im-Jeong Choi, Su Young Hong, Yu-Mi Kim, Jae-Ho Yoo Kosin University, College of Medicine, Busan, Korea; Good Gang-An Hospital, Busan, Korea; Pusan National University, Children[apos]s Hospital, Yangsan, Korea; Wallace Memorial Baptist Hospital, Busan, Korea; Inje University, College of Medicine, Busan, Korea; Maryknoll Medical Center, Busan, Korea; Dong-A University, College of Medicine, Busan, Korea; Dong-A University, College of Medicine, Busan, Korea Objective: We performed this study to investigate changes in the incidence of type 1 diabetes mellitus(T1DM) and type 2 diabetes mellitus(T2DM) in children aged below 15 years in Busan and Gyeongnam province, Korea during the period from 2001 to 2010.[br]Methods: We sent questionnaires via the post to 5 tertiary and 42 general hospitals in Busan and Gyeongnam province. We received answered questionnaires based on medical records from all tertiary and 11 general hospitals. Three hundred forty four diabetic patients(239 T1DM, 89 T2DM, 16 unclassified DM) who were newly diagnosed from 2001 to 2010 were enrolled. The incidence rates were calculated as the number of cases per 100,000 person-years. The denominator for incidence rate was secondary data from the population registry(Korea National Statistical Office). The incidence rate was fitted and test for the linear trend was done by Poisson linear regression model. We also used the Poisson regressions to assess the rate ratio for development of diabetes with year, sex, age group, geographic location.[br]Results: The average crude incidence was 2.01(95% CI: 1.77-2.28) and 0.76(95% CI: 0.61-0.93) for T1DM and T2DM, respectively. There was a significant increasing trend in the incidence of T1DM during the period of 2001 and 2010, with an annual 1.08-fold increase of the rate ratio(95% CI: 1.03-1.12). The trend in the incidence of T2DM increased with an annual 1.23-fold increase of the rate ratio(95% CI: 1.14-1.33). The incidence of T2DM among those aged 10-14 years was rapidly increased during the period of 2001 to 2010 and was higher than that of T1DM in 2010.[br]The rate ratio for development of T1DM was 1.16-fold higher in males than females and that for those aged 10-14 years was a 2.04-fold higher when compared to those aged 0-4 years. Also, there was an 1.43 increased rate ratio in Busan(urban) compared to Gyeongnam province(suburban). The rate ratio for development of T2DM was 1.29-fold higher in males than females and that for those aged 10-14 years was a 9.59-fold higher when compared to those aged 5-9 years. Also, there was an 1.58 increased rate ratio in Busan(urban) compared to Gyeongnam province(suburban).[br]Conclusions: The incidence of T1DM and T2DM have shown a significant increasing trend for last 10 years in Busan and Gyeongnam province, Korea. Our study shows the needs for careful monitoring of incidence and its related risk factors of T2DM in adolescents.[br][br]Nothing to Disclose: JHL, MJK, SYK, H-JK, SHL, CC, WYC, I-JC, SYH, Y-MK, J-HY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1467 181 1252 SUN-195 PO12-01 Sunday 1047 2012


1046 ENDO12L_SUN-196 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Genomic Health Disparities, Type 2 Diabetes and Mexican-Americans Liliana Uribe-Bruce, Maria Isabel Garcia, Monica Ruiz, Linda Gallo, Johanna Euyoque, Patricia Gonzalez, Addie Brewer Fortmann, Athena Philis-Tsimikas The Scripps Research Institute, La Jolla, CA; The Scripps Whittier Diabetes Institute, La Jolla, CA; San Diego State University, San Diego, CA [bold]Background: [/bold]Type 2 Diabetes (T2D) is a worldwide epidemic disproportionately burdening certain racial and ethnic groups. In the U.S, the prevalence of T2D is 12% in Hispanics, compared to 7% in non-Hispanic whites (1). The epidemic is due to lifestyle, environment, genetic and epigenetic factors. Although over 40 genetic risk markers for T2D have been identified, 95% of studies have been conducted exclusively in European-origin populations, creating a genomics health gap. (2)[br][bold]Project Aims: [/bold] To elucidate genomic factors potentially related to the disproportionate risk for T2D among Hispanics, by fostering participation of this group in genomics research, and by creating a dedicated Genebank to allow this research.[br][bold]Methods: [/bold]A community engaged research approach is being used to identify factors responsible for low participation in genomics research among medically underserved Mexican-Americans. Community response to recruitment effort is recorded. To create awareness and ensure informed consent when enrolling in the Genebank, a culturally and literacy-tailored educational tool has been developed and delivered in Spanish at community clinics. IRB approved community focus groups discussing knowledge and attitudes towards genomic research have been held, and a survey has been launched. The [quot]Scripps San Diego Diabetes Genebank[quot] is actively enrolling volunteers.[br][bold]Results: [/bold] Close to 300 volunteers have already enrolled in the Genebank. About 70% are female, ages from 21 to 81 years old. 65% of those referred as candidates to participate in the Genebank have enrolled. 10% were excluded due to study regulations, 8% lost contact and 17% declined participation. Reasons to decline participation are: 13% lack of time or transportation, 7% fear of blood draws, 3% lack of perceived benefit, and 3% distrust for research. FOCUS GROUPS: Over 90% of participants have expressed interest in learning about their genomic makeup and allowing healthcare providers to use genomics to treat them or future generations. 35% have expressed concern about genomics research tampering with nature.[br][bold]Conclusions: [/bold] Medically underserved Hispanics have positively responded and participated in genomics research in T2D, and expressed interest in learning about their genetic makeup, being approached by invitation that includes a culturally and literacy tailored education tool. Only 3% have expressed distrust for researchers, and 100% mention altruism as a reason to participate.[br][br](1) CDC-2011 National Diabetes Fact Sheet. Accessed through the web on February 2012 at http://www.cdc.gov/diabetes/pubs/estimates11.htm#. (2) Bustamante C., et al. Genomics for the world. Nature, July 2011, vol. 475, p163.[br][br]Sources of Research Support: NIH CTSA grant UL1 RR025774.[br][br]Nothing to Disclose: LU-B, MIG, MR, LG, JE, PG, ABF, AP-T 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1813 181 1253 SUN-196 PO12-01 Sunday 1048 2012


1047 ENDO12L_SUN-197 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Obesity, Anxiety and Depression in Outpatients with Type 2 Diabetes in a Mexican Population Fatima Cristhel Martinez Hernandez, Samantha Solis Vidal, Lilia Lopez Narvaez, Isela Juarez Rojop, Carlos Alfonso Tovilla Zarate, Teresa Ramon Frias, Deysi Yadira Bermudez Ocana, Jorge Eduardo Reyes Tovilla Universidad Ju[aacute]rez Aut[oacute]noma de Tabasco, Comalcalco, Mexico; CIGEN, Centro de Investigaci[oacute]n Gen[oacute]mica, Comalcalco, Mexico; Universidad Ju[aacute]rez Aut[oacute]noma de Tabasco, Villahermosa, Mexico Background: Obesity is a very important factor for the development of depression and anxiety. There is evidence indicating a relationship between depression and anxiety in diabetic patients. However, the relationship between obesity and depression and anxiety in diabetes has been poorly analyzed. The aim of this study was to assess the prevalence of anxiety and depression in groups of obese and normal-weight individuals with type 2 diabetes and to analyze the severity of depression in type 2 diabetics when taking into account gender, normal-weight and obese factors in people with this condition.[br]Methods: This study included 467 patients with type 2 diabetes, 236 with normal weight and 231 with obesity. All participants completed a demographic questionnaire, the Hamilton Anxiety Rating Scale and the Hamilton Depression Rating Scale. Anthropometric characteristics were applied. To analyze the data, the Mann-Whitney U-test was employed.[br]Results: In patients with obesity, 48.48% (95% CI: 41.82-55.14) were positive for anxiety and 49.78% (95% CI: 43.11-56.44) for depression. With regard to depression symptoms, a significant difference between normal-weight and obese individuals with type 2 diabetes (p=0.05) was found. Also, when we analyzed depression scores, significant differences were encountered for gender between normal-weight and obesity groups in the range of severe (p=0.01) and very severe (p=0.04) cases. Significant differences were not observed when we analyzed for anxiety.[br]Conclusion: In view of the existing relationships observed among obesity, depression, anxiety and gender in type 2 diabetics, a psychological intervention is necessary for an integral managing of these patients.[br][br]Sources of Research Support: Data collection was accomplished thanks to the support of grant P.O.A. 20111282 from UJAT-DAMC.[br][br]Nothing to Disclose: FCMH, SSV, LLN, IJR, CATZ, TRF, DYBO, JERT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 952 181 1254 SUN-197 PO12-01 Sunday 1049 2012


1048 ENDO12L_SUN-198 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) The EURO-WABB Project: An EU Register for Wolfram, Alstr[ouml]m, Bardet Biedl and Other Rare Diabetes Syndromes Pietro Maffei, Amy Farmer, Gabriella Milan, Francesca Favaretto, Vera Bettini, Segolene Ayme, Miguel Lopez de Heredia, Susan Mc Cafferty, Wojciech Mlynarski, Virginia Nunes, Kay Parkinson, Veronique Paquis, Richard Sinnott, Vallo Tillman, Roberto Vettor, Tim Barrett Padua University Hospital, Padua, Italy; Birmingham Children[apos]s Hospital, Birmingham, UK; INSERM, Paris, France; IDIBELL, Barcelona, Spain; University of Glasgow, Glasgow, UK; Medical University of Lodz, Lodz, Poland; Alstrom Syndrome UK, Paignton, UK; CNRS, Nice, France; University of Tartu, Tartu, Estonia; University of Birmingham, Birmingham, UK; CIBERER, Barcelona, Spain; UB, Barcelona, Spain Introduction: EURO-WABB is a European research project within the field of rare diabetes diseases. The general objective is to support efficient diagnosis, treatment and research for the overlapping rare genetic diseases Wolfram, Alstrom and Bardet-Biedl (WABB) syndromes. Methods: The project is supported by the EU DG-SANCO by the collaboration of 8 Associated Partners (AP) and 15 Collaborating Partners. University of Birmingham works as the Project Leader (TB) and coordinating centre. A web-platform was launched in 2011 (www.euro-wabb.org) and a virtual registry will be soon available. Results: Key achievements of the project are yet on the ground and include: 1) coordination: Project Management and Scientific Advisory Committee established; 2) Dissemination: awareness raising through conference attendance and development of multi-language website; 3) Evaluation: regular conference calls with AP to monitor project progress; 4) Core datasets: core and extended dataset agreed in September 2011; 44 + 370 data fields respectively; phenotyping information standardised used ICD-10 and ESPE classification coding systems; 5) Genetics: mutation database completed for ALMS1, WFS1, SLC19A2 and EIF2AK3; 6) Virtual Registry and Information Environment: prototype core dataset registry developed for testing in November 2011. First recruitment of patients started in August 2011 and local Ethical Committee approval was obtained in UK, Italy, Spain and Poland. In the period from 20th June-2nd January 2012, 2814 visits from 73 countries accessed the website (2295 from EU). Up to date, 22 people have formally registered including a mix of patients, families, clinicians and other interested parties. We currently have the core dataset for 29 patients in the online registry Conclusion: EURO-WABB is a new reality in the world of rare diseases. By the virtual registry and the website it will be possible in the next future to address the main topics of this project. Next milestones will include the publication of consensus management guidelines, diagnostic criteria, health professional education and patient information material to support the registry.[br][br]Sources of Research Support: EURO-WABB; agreement number-20101205.[br][br]Nothing to Disclose: PM, AF, GM, FF, VB, SA, MLdH, SMC, WM, VN, KP, VP, RS, VT, RV , TB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 815 181 1255 SUN-198 PO12-01 Sunday 1050 2012


1049 ENDO12L_SUN-199 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Pre-Operative Glycemic Control and Hospital Outcomes in Patients with Diabetes Undergoing Low to Moderate Risk Elective Surgical Procedures Ali R Shoraka, Aashish A Shah, Colleen S Thomas, Carl A Ruthman, Melanie M Thomas, Joan Irizarry Alvarado, Gunjan Y Gandhi Mayo Clinic, Jacksonville, FL; Mayo Clinic, Jacksonville, FL [bold]Background:[/bold] Poor glycemic control prior to coronary artery bypass grafting has been associated with increased risk of complications. Although frequently assumed, there is paucity of data regarding hyperglycemia as a predictor of adverse events in low to moderate risk surgeries.[br][bold]Objective:[/bold] To estimate the magnitude of association between pre-operative glucose control (as measured by HbA1C) and post-operative outcomes in surgical patients with diabetes.[br][bold]Methods:[/bold] Single center prospective observational study of consecutive adult patients with diabetes seen at a pre-operative evaluation clinic. For primary analysis, the association of HbA1C with the primary composite endpoint of various clinically important outcomes was evaluated using a multivariable logistic regression model adjusted for age, gender, diabetes type, body-mass index, smoking status and type of surgery.[br][bold]Results:[/bold] Of 431 patients who met inclusion criteria, six were excluded from analyses, as they did not have HbA1C measured. The median age was 67 years, 53% were male, more than half were obese (median BMI: 31 kg/m[sup]2[/sup]) and 94% had type II diabetes. Orthopedic (23%), general (19%) and urologic (12%) surgical procedures were most commonly performed. 89 (21%) patients experienced at least one complication. Renal insufficiency (8%), new-onset atrial fibrillation (7%), delirium (4%) and infections (4%) were amongst the most common. Of the 127 patients with suboptimal glycemic control (HbA1C [gt]7%), 34 (27%) experienced complications compared to 55 (18%) among 298 with good glycemic control (HbA1C [underline][lt] [/underline]7%). Those with HbA1C[gt]7% were younger (median age: 64 vs. 68 yrs), more likely to be treated with insulin only preoperatively (39% vs. 16%), have type I diabetes (9% vs.2%), associated coronary artery disease (33% vs.21%), and a higher median pre-operative fasting glucose (179 vs. 112 mg/dL). After multivariable adjustment, suboptimal pre-operative HbA1C was associated with an estimated 88% increase in odds of experiencing one or more adverse outcomes (OR 1.88, 95% CI 1.09-3.22, P=0.023) but not with a longer hospital stay (median 4 days, p=0.75). Preoperative plasma glucose [gt]200 mg/dL was associated with a non-significant increased risk of complications (adjusted OR 1.47, 95% CI 0.73-2.98).[br][bold]Conclusion:[/bold] In patients with diabetes undergoing low to moderate risk elective surgeries, pre-operative HbA1C is an independent predictor of adverse patient-important outcomes.[br][br]Sources of Research Support: Mayo Clinic Institutional Grant.[br][br]Nothing to Disclose: ARS, AAS, CST, CAR, MMT, JIA, GYG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 514 181 1256 SUN-199 PO12-01 Sunday 1051 2012


1050 ENDO12L_SUN-200 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Screening for Diabetes: Can Simpler Tests Substitute for the Oral Glucose Tolerance Test (OGTT)? Darin E Olson, Jessica Vakili, Ranee Chatterjee, Qi Long, Lawrence S Phillips Atlanta VAMC, Decatur, GA; Emory University School of Medicine, Atlanta, GA; Rollins School of Public Health Emory University, Atlanta, GA; Atlanta VAMC, Decatur, GA; Duke University, Durham, NC The OGTT is the standard to identify diabetes with maximum sensitivity and specificity, but is inconvenient and time-consuming. We asked if combinations of simpler tests could substitute for the OGTT. In the Screening for Impaired Glucose Tolerance study, 1573 adults without known diabetes had random plasma glucose (RPG), A1c, and a simplified 50g oral glucose challenge test (GCT, with plasma glucose after 1 hour) [ndash] all tests that can be done opportunistically, at any time of day, without a prior fast, during outpatient visits. OGTT found 4.6% to have previously unrecognized diabetes. The accuracy of screening to identify subjects with OGTT-diabetes was assessed by the area under curves (AUC or c-statistic) in receiver operating characteristic analysis. FPG (from the OGTT) and GCT had the highest AUC as single tests, 0.93 and 0.90, respectively (p=ns), both greater than RPG and A1c (both p[lt]0.02). Combining any two tests improved AUC vs. the component tests (p[lt]0.01 for all), and FPG+GCT had the highest AUC (0.96, p[lt]0.03 vs. all other combined tests). The greatest sum of sensitivity and specificity was provided by the cutoffs of GCT[ge]120 and FPG[ge]110 (sensitivity 81%, specificity 94%). Instead of using an OGTT, GCT could be done conveniently during an outpatient visit, followed by an FPG in patients with GCT [ge]120. If the 9.5% of patients who failed both GCT and FPG cutoffs were to be defined as having diabetes, that would include 5.8% false positives and 0.9% false negatives compared to the OGTT. We examined the cost impact of such discrepancies, by projecting healthcare and societal costs for screening and 3 years of treatment with metformin (including costs for false positives and false negatives), for those defined as having diabetes by GCT+FPG vs. use of an OGTT. Dual screening with GCT+FPG, without an OGTT, would produce cost savings vs. no screening [ndash] but cost savings would be even greater for a single screen with the GCT, followed if positive by an OGTT. [underline]Conclusion[/underline]: Combinations of simple tests improve screening accuracy, and the best combination tested was FPG+GCT. However, it is more cost-effective to screen for diabetes with a 50g oral glucose challenge test with measurement of glucose 1 hr later (at any time of day, without regard to meals), followed, if positive, by an OGTT, compared to screening without an OGTT and using only combinations of other tests such as fasting plasma glucose, random plasma glucose, or A1c.[br][br]Sources of Research Support: VA HSR[amp]D Award IIR 07-13.[br][br]Disclosures: LSP: Investigator, Novo Nordisk, Merck & Co., Amylin Pharmaceuticals, Sanofi-Aventis; Advisory Group Member, Boehringer Ingelheim. Nothing to Disclose: DEO, JV, RC, QL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1685 181 1257 SUN-200 PO12-01 Sunday 1052 2012


1051 ENDO12L_SUN-201 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Prevalence of Tinel Sign in Patients with Diabetes Isaam Cheikh, Shahreyar Shar Hashemi, Arnold Lee Dellon Union Memorial Hospital, Baltimore, MD; Dellon Institute for Peripheral Nerve Surgery, Towson, MD [bold]OBJECTIVE: [/bold]To evaluate the prevalence of the Tinel Sign in the upper and lower extremities of patients with diabetes in an urban community.[br][bold]RESEARCH DESIGN AND METHODS:[/bold] An IRB-approved, prospective, cross-sectional, descriptive study was performed on patients with diabetes during their visit to the endocrinologist. Eighty-one consecutive patients (162 sides) were examined by a fellowship-trained Hand Surgeon experienced in performing the [ldquo]Tinel Sign[rdquo]. The number of Type I and II diabetes were one and eighty, respectively. The mean duration of diabetes was 18.5 years. In the upper extremity the known sites of anatomic entrapment were percussed: median nerve at the wrist (carpal tunnel), ulnar nerve at the elbow (cubital tunnel), radial sensory nerve in the forearm, and a negative control site. In the lower extremity, the sites were the tibial nerve at the ankle (tarsal tunnel), proximal tibial nerve (soleal sling), the common peroneal nerve at the fibular head (fibular tunnel), superficial peroneal nerve, deep peroneal nerve over dorsum of the foot, and a negative control site. Neuropathy was defined as a score greater than 4 on the Michigan Neuropathy Screening Instrument (MNSI). 95% confidence intervals were utilized for each prevalence. Statistical evaluation included linear regression, analysis of variance, and Tukey[apos]s post hoc test.[br][bold]RESULTS: [/bold]The Michigan Neuropathy Score and the number of positive Tinel Signs were highly correlated (coefficient r=0.94, p=0.001) in the patients with diabetes that were tested. In the upper extremity, the prevalence of Tinel Sign in patients with neuropathy (i.e. MNSI [gt]4) ranged from 46.2% to 65% compared to 2.4% to 19.5% without neuropathy (MNSI [le] 4). Lower extremity prevalence ranges were 23% to 59% with neuropathy and 0% to 17.1% without neuropathy. The chances of having a positive Tinel Sign in the contralateral limb were 38.2%, 42.2%, 44.7% and 48.4% for carpal tunnel, cubital tunnel, the fibular tunnel and tarsal tunnel, respectively. All of the control test sites had an absent Tinel Sign (negative Tinel).[br][bold]CONCLUSION: [/bold]Chronic nerve compression in patients with diabetes, as assessed by the presence of Tinel Signs at eighteen anatomic sites, was documented. Prevalence quadrupled in the presence of neuropathy.[br][br]Nothing to Disclose: IC, SSH, ALD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 105 181 1258 SUN-201 PO12-01 Sunday 1053 2012


1052 ENDO12L_SUN-202 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) N-Acetylcysteine (NAC) Prevents the Impairment of the Counter-Regulatory Response Following Recurrent Hypoglycemia Adam Deak, Xavier Fioramonti, Shrinidi Deshpande, Corinne Leloup, Nazish Sayed, Luc Penicaud, Annie Beuve, Vanessa H Routh University of Medicine and Dentistry of New Jersey, Newark, NJ; University of Dijon, Dijon, France; Stanford University, Stanford, CA Hypoglycemia is a severe side effect of intensive insulin therapy. Recurrent hypoglycemia (RH) impairs the counter-regulatory response (CRR) which restores euglycemia. The ventromedial hypothalamus (VMH) detects hypoglycemia and initiates the CRR. VMH nitric oxide (NO) production and activation of it[apos]s receptor soluble guanylyl cyclase (sGC) is necessary for the CRR. When NO is produced in the presence of reactive oxygen species (ROS) protein S-nitrosylation occurs. S-nitrosylation of sGC impairs its function and desensitizes NO signaling. We hypothesize that during hypoglycemia, the interaction between NO and ROS increases S-nitrosylation levels. This reduces NO activation of sGC and impairs the CRR. In support of this, insulin-hypoglycemia increases VMH ROS levels by 49.66 [plusmn] 18.37% (P[lt]0.05). Moreover, 3 consecutive daily episodes of insulin-hypoglycemia (RH model) increase VMH sGC S-nitrosylation. We then determined whether preventing ROS production, and consequently S-nitrosylation of sGC prevents the impaired CRR after RH by treating rats with the antioxidant N-acetylcysteine (NAC) in their drinking water (5 g/l) for 9 days before and during RH (NAC pre-treatment). After RH, glucose levels fell further and epinephrine production was reduced by 50% in response to insulin-hypoglycemia compared to controls (glucose nadir RH: 39.3 [plusmn] 1.1 mg/dl; single hypoglycemia [SH]: 53.5 [plusmn] 1.8 mg/dl; epinephrine 120 min RH: 611.2 [plusmn] 108.3 ng/l; SH: 1350.6 [plusmn] 95 ng/l; p[lt]0.05). After NAC pre-treatment there were no significant differences in glucose nadir between RH and SH animals (glucose nadir RH + NAC: 39.6 [plusmn] 3 mg/dl; SH + NAC: 35 [plusmn] 3 mg/dl). Moreover, after NAC pre-treatment the RH epinephrine response was restored to that of the SH group (RH + NAC: 1390 [plusmn] 148.4 ng/l vs SH, p[gt]0.05). NAC also reversed sCG S-nitrosylation. Next we determined whether NAC reverses the impaired CRR by injecting NAC (200mg/kg; i.p). into RH rats 4 hours after the 3[sup]rd[/sup] episode of insulin hypoglycemia. NAC significantly increased glucagon production following RH (RH+NAC: 140 [plusmn] 9.2 vs RH: 102.4 [plusmn] 8.4; p[lt]0.05). Interestingly, following RH insulin-hypoglycemia produced no further increase in ROS production suggesting that ROS[ndash]induced S-nitrosylation is sustained in the absence of further ROS production. These data suggest that NAC prevention of ROS production during hypoglycemia may be clinically useful in preventing impaired CRR in patients undergoing intensive-insulin therapy.[br][br]Sources of Research Support: DK81538, Juvenile Diabetes Research Foundation;. P41 RR001395 (NCRR).[br][br]Nothing to Disclose: AD, XF, SD, CL, NS, LP, AB, VHR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2106 181 1259 SUN-202 PO12-01 Sunday 1054 2012


1053 ENDO12L_SUN-203 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Exercise Increases Serum Fibroblast Growth Factor 21 (FGF21) Levels Daniel Cuevas-Ramos, Paloma Almeda-Valdes, Francisco J Gomez-Perez, Clara E Meza-Arana, Griselda Brito-Cordova, Jorgue Oseguera-Moguel, Carlos A Aguilar-Salinas Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Mexico City, Mexico; Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Mexico City, Mexico; Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Mexico City, Mexico [bold]Introduction: [/bold]Fibroblast growth factor 21 (FGF21) increases glucose uptake. In a cross-sectional study, FGF21 was associated with daily physical activity (1). It is unknown if FGF21 serum levels are affected by exercise. [bold]Methods: [/bold]This was a comparative and longitudinal study. Anthropometric and biochemical evaluations were done before and after a bout of exercise and repeated after two weeks of daily supervised exercise. The study sample was composed by 60 sedentary young healthy women. [bold]Results: [/bold]The mean age was 24[plusmn]3.7 years old, with a mean BMI of 21.4[plusmn]7.0 kg/m[sup]2[/sup]. The anthropometric characteristics did not change after two weeks of exercise. Serum FGF21 levels increased significantly after two weeks of exercise [276.8 ng/l (142.8-568.6) vs (460.8 (298.2-742.1), p[lt]0.0001). A significant reduction of leptin (13.1 ng/ml (8.2-21.1) vs 12.4 (10.0-16.2), p=0.001) was also identified. The delta (final[ndash]basal) log of serum FGF21, adjusted for BMI, had significant positive correlation with basal glucose (r=0.23, p=0.04), mean maximal heart rate (MHR) (r=0.54, p[lt]0.0001), mean METs (r=0.40, p=0.002), delta plasma epinephrine (r=0.53, p[lt]0.0001) and delta plasma FFAs (r=0.35, p=0.006). Stepwise linear regression model showed that glucose, MHR, METs, FFAs, and epinephrine, were factors independently associated with the increment of FGF21 after exercise program (F=4.32; r[sup]2[/sup]=0.64, p[lt]0.0001). [bold]Conclusions: [/bold]Serum FGF21 significantly increased after two weeks of physical activity. This increment correlated positively with clinical and biochemical parameters related with adrenergic and lipolytic response.[br][br]Cuevas-Ramos D et al., Eur J Endocrinol 2010;163:469.[br][br]Nothing to Disclose: DC-R, PA-V, FJG-P, CEM-A, GB-C, JO-M, CAA-S 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 110 181 1260 SUN-203 PO12-01 Sunday 1055 2012


1054 ENDO12L_SUN-204 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Cardiorespiratory Fitness Is Decreased and Not Correlated with C-Reactive Protein Levels in Adults with Diabetes Mellitus Cristina O Francisco, Aparecida M Catai, Silvia C Moura, Sergio L Lopes, Adriano M Del Vale, Angela M Leal Federal University of S[atilde]o Carlos, S[atilde]o Carlos, Brazil; Federal University of S[atilde]o Carlos, S[atilde]o Carlos, Brazil Diabetes Mellitus (DM) has been associated with lung dysfunction and impairment of cardiorespiratory fitness and increasing evidences have suggested that comorbidities and systemic inflammation may be involved in these derangements. However, the results regarding cardiopulmonary dysfunction and its underlying mechanisms in DM are still controversial. In this study, we evaluated the relationship between metabolic variables and inflammation marker, including hemoglobin A1C (HbA1C), lipid profile and C-reactive protein (CRP) levels, and cardiorespiratory fitness in adult patients with DM. Nineteen men with diabetes (aged 51 [plusmn] 6 years; mean [plusmn] SD) and nineteen age and sex-matched control subjects (aged 49 [plusmn] 7 years; mean [plusmn] SD) were studied. The average duration of diabetes was 11 years (range, 2-25 years) and the HbA1C mean levels were 8.4% (range, 6-12.9%). All individuals were subjected to incremental cardiopulmonary exercise test and spirometry. Blood pressure, triglycerides, HDL, LDL, total cholesterol and CRP were not different in control subjects and individuals with DM. Body mass index (28.7 [plusmn] 1 vs 25.6 [plusmn]0.4 kg/m2, mean [plusmn] SD, p[lt]0.01) and resting heart rate (78.3 [plusmn] 2.2 vs 71.3 [plusmn] 2.1 beats/min, mean [plusmn] SD, p=0.03) were significantly higher in individuals with DM than in control subjects, respectively. No significant difference was noted in spirometric variables, including forced vital capacity (FVC), forced expiratory volume in 1s (FEV1), forced expiratory flow midexpiratory phase (FEV25-75%) and peak expiratory flow (PEF). In the exercise test, maximal overload (W), peak heart rate (HRpeak), peak oxygen consumption (VO2peak) at anaerobic threshold (VO2AT) and respiratory exchange ratio (RER) were significantly lower in patients with diabetes than in control subjects. No correlations were observed between HbA1C and CRP plasma levels and cardiorespiratory variables. These data indicate that cardiorespiratory fitness is reduced and spirometric values are preserved in patients with diabetes. However, decreased cardiorespiratory fitness is not correlated with CRP levels or HbA1C.[br][br]Sources of Research Support: FAPESP.[br][br]Nothing to Disclose: COF, AMC, SCM, SLL, AMDV, AML 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 35 181 1261 SUN-204 PO12-01 Sunday 1056 2012


1055 ENDO12L_SUN-206 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Renal and Metabolic Biomarker and Efficacy Effects of Bardoxolone in Uninephrectomized [italic]db/db[/italic] Mice Keyun Qing, Kathleen Heinz-Taheny, James V Ficorilli, Donalyn Scheuner, Yanping Xu, Timothy Grese, Matthew D Breyer, Mark C Kowala, Laura F Michael Eli Lilly and Company, Indianapolis, IN According to the Centers for Disease Control and Prevention, more than 10% of the US population older than 20 years old suffered chronic kidney disease (CKD) in 2010. While therapeutic options exist for treatment of risk factors that lead to CKD, such as anti-hypertensive and anti-diabetic agents, these treatments are suboptimal as patients continue to progress to end stage renal disease (ESRD). Bardoxolone is a novel anti-oxidant therapeutic that is in late clinical trial development for the prevention of end stage renal disease in patients with Stage 4 CKD and type 2 diabetes mellitus (T2DM). The goal of our study was to quantitate bardoxolone-mediated effects on renal function and pathology and improvements in metabolic parameters that are prevalent in patients with CKD and T2DM using the [italic]db/db[/italic] mouse model. Four weeks following uninephrectomy, mice were dosed orally with 1, 3, or 10 mg/kg bardoxolone or 5 mg/kg rosiglitazone daily for 8 weeks. Renal expression of the anti-oxidative stress gene, [italic]Nqo1[/italic], was induced dose-responsively by bardoxolone but not by rosiglitazone. Rosiglitazone induced body weight gain; whereas, bardoxolone had no effect at any dose level. Fed blood glucose and urinary glucose levels decreased following rosiglitazone and dose-dependently decreased in response to bardoxolone. Blood urea nitrogen did not change; however, serum creatinine was slightly reduced in both bardoxolone and rosiglitazone treated groups. Urinary albumin-creatinine ratio was reduced significantly by rosiglitazone but not by bardoxolone. Histopathology showed a marginal trend for dose-dependent decrease in mesangial matrix expansion in mice treated with bardoxolone. Periglomerular/glomerular fibrosis were dose dependently decreased in the bardoxolone-treated mice, and glomerular necrosis severity scores and incidence was dose-dependently decreased in the bardoxolone-treated mice where all dose groups had average scores lower than the vehicle-treated mice. Accordingly, renal expression of TGFb, a growth factor associated with kidney fibrotic disease, was reduced in mice treated with bardoxolone and rosiglitazone. In total, these data illustrate improvement in glucose control in the rodent model of T2DM and suggest a possible improvement in glomerular structure following bardoxolone treatment. Whether the bardoxolone-associated improvement in renal pathology is a result of improved glucose control or a result of reduced oxidative stress in the kidney is unknown.[br][br]Nothing to Disclose: KQ, KH-T, JVF, DS, YX, TG, MDB, MCK, LFM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1692 181 1262 SUN-206 PO12-01 Sunday 1058 2012


1056 ENDO12L_SUN-207 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) DPP4 Levels in Peripheral Blood and T Cells in Patients with Type 2 Diabetes Sang Ah Lee, Youngree Kim, Sun Hyung Kim, Sung Ha Kang, Jinseok Kim, Sang Taek Heo, Gwanpyo Koh, Dae Ho Lee Jeju National University School of Medicine and Jeju National University Hospital, Jeju, Korea; Jeju National University School of Medicine and Jeju National University Hospital, Jeju, Korea CD26, known as dipeptidyl peptidase 4, is a multifunctional protein which is expressed on various cells including T cells and also circulates in a soluble form. However, the clinical implications of CD26 expressed on T cells and circulating soluble CD26 (sCD26) in patients with type 2 diabetes (T2DM) are not clarified yet. We evaluated the association between those CD26s and clinical parameters in patients with T2DM (n=109). Exclusion criteria were as follows: patients with systemic infection, hepatic or renal disease, or other severe systemic illnesses, patient on DPP4 inhibitor medication. CD4+CD26+ and CD8+CD26+ T cell fractions and sCD26 were measured by using a flow cytometer and an ELISA kit, respectively. Compared with healthy control subjects (n=14), T2DM group had increased the fractions of CD4+CD26+ (11.2% vs. 19.6%, [italic]p[/italic][lt]0.01) and CD8+CD26+ T cells (3.8% vs. 6.6%, [italic]p[/italic][lt]0.01) in peripheral blood. HbA1c was only one independent factor associated with CD4+CD26+ and CD8+CD26+ T cell fractions in a multivariate analysis ([beta]=0.309, [italic]p[/italic][lt]0.01 and [beta]=0.233, [italic]p[/italic]=0.04). However, serum sCD26 level was lower in patients with T2DM compared with the control group (85.6 vs. 111.8 ng/dL, [italic]p[/italic][lt]0.05). Serum DPP4 activity was comparable between the groups. After active glucose control for 8 weeks or more in drug na[iuml]ve T2DM patients (n=37), the fraction of CD4+CD26+ T cell, but not CD8+CD26+ T cell, was decreased significantly. Our results suggest that CD26 level on T cells, but not sCD26 and DDP4 activity, in peripheral blood was associated with sugar control status in patients with T2DM.[br][br]Nothing to Disclose: SAL, YK, SHK, SHK, JK, STH, GK, DHL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1405 181 1263 SUN-207 PO12-01 Sunday 1059 2012


1057 ENDO12L_SUN-208 POSTER SESSION: Diabetes [amp] Its Complications: Epidemiology, Genetics, [amp] Pathophysiology (1:30 PM-3:30 PM) Circulating CART Levels in Patients with Diabetes Mellitus Amir H Sam, Paul Bech, Anjali Amin, Radha Ramachandran, Niamh M Martin, Mohammad A Ghatei, Stephen R Bloom, Kevin G Murphy Imperial College London, London, UK Background: Cocaine- and amphetamine-regulated transcript (CART) codes for a peptide expressed in nervous and endocrine tissues. CART expression is upregulated in the pancreatic islet beta-cells in several rodent models of type 2 diabetes. We aimed to determine whether circulating CART levels are different in patient with diabetes mellitus.[br]Methods: Plasma CART levels were measured in patients with type 2 diabetes (n=31), type 1 diabetes (n=22) and controls (n=23) using an in-house radioimmunoassay.[br]Results: Plasma CART levels were not different in the three groups (Mean[plusmn]SEM: type 1 diabetes mellitus: 81.98 [plusmn] 6.82 pmol/L, type 2 diabetes mellitus: 82.02 [plusmn] 8.05 pmol/L, control: 61.28 [plusmn] 8.76 pmol/L, p= 0.13).[br]Conclusions: Circulating CART levels are unlikely to contribute to the pathogenesis of diabetes mellitus in humans. However, paracrine effects of CART in the pancreas cannot be excluded.[br][br]Nothing to Disclose: AHS, PB, AA, RR, NMM, MAG, SRB, KGM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2250 181 1264 SUN-208 PO12-01 Sunday 1060 2012


1076 ENDO12L_SUN-229 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Effects of Increased [beta][sub]2[/sub]-Adrenergic Receptor Signaling on Hepatic Steatosis during Aging Bin-Xian Zhang, Zhen-Ju Shu, Chih-Ko Yeh, Amrita Kamat University of Texas Health Science Center at San Antonio, San Antonio, TX; Audie Murphy Veterans Hospital, San Antonio, TX; University of Texas Health Science Center at San Antonio, San Antonio, TX Fatty liver has emerged as a risk factor contributing substantially to the pathophysiology of insulin resistance, type 2 diabetes, nonalcoholic fatty liver disease, and cardiovascular diseases. Age has been demonstrated to be an independent risk factor for increased hepatic fat accumulation. However, the precise mechanisms triggering fat build-up in the liver during aging are still unclear. Advancing age enhances the release of catecholamines which act via beta-adrenergic receptors ([beta][sub]1[/sub]-, [beta][sub]2[/sub]- and [beta][sub]3[/sub]-AR subtypes) to regulate lipid and glucose metabolism. Earlier studies have demonstrated an age-associated increase in hepatic [beta]-AR membrane content and associated adenylyl cyclase activity, a phenomenon unique to the liver. We have previously shown that [italic]in vitro[/italic] overexpression of [beta][sub]2[/sub]-ARs in hepatocytes from young rodents increases lipid content in the cells, whereas inhibition of [beta][sub]2[/sub]-ARs by receptor subtype-specific agonist ICI118551 reduces lipid levels in hepatocytes from senescent animals. In the current studies, we have observed that increases in hepatic lipid accumulation upon administration of [beta]-AR agonist isoproterenol to young mice [italic]in vivo[/italic] is prevented by propranolol, a beta blocker, indicating a novel effect of this class of drugs in hepatic steatosis. On the other hand, Acipimox, which inhibits adipose tissue lipolysis and reduces hepatic free fatty acid uptake, does not alter isoproterenol-mediated hepatic fat accumulation. These results suggest that mechanisms intrinsic to the liver are involved in increasing hepatic lipid accumulation upon isoproterenol administration. Further studies with [beta][sub]2[/sub]-AR knockout mice have shown that ablation of [beta][sub]2[/sub]-AR reduces age-associated increase in hepatic fat accumulation and lipid peroxidation, improves glucose tolerance and increases insulin sensitivity. A number of factors including increased de novo lipogenesis, decreased beta-oxidation, reduced triglyceride secretion or a combination of these factors could contribute to [beta][sub]2[/sub]-AR-mediated lipid accumulation in liver. Studies are currently in progress using wild-type and [beta][sub]2[/sub]-AR knockout mice to evaluate the mechanisms involved in [beta][sub]2[/sub]-AR-mediated hepatic steatosis during aging.[br][br]Sources of Research Support: American Heart Association Grant-in-Aid Award; Kronos Longevity Research Institute; CTSA grant (UL1RR025767).[br][br]Nothing to Disclose: B-XZ, Z-JS, C-KY, AK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 80 183 1283 SUN-229 PO08-01 Sunday 1079 2012


1077 ENDO12L_SUN-230 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Female-Specific ER[alpha] Activity Protects Against Sequelae of High-Fat Feeding: Weight Gain, Glucose Tolerance, and Fatty Liver Disease Sarah Hart-Unger, Katherine Hamilton, Michael Freemark, Kenneth Korach Duke University Medical Center, Durham, NC; National Institute of Environmental Health Sciences, Research Triangle Park, NC Nonalcoholic fatty liver disease (NAFLD) is defined as the accumulation of excess lipid in the liver in the absence of excess alcohol consumption. The risk of NAFLD is increased in patients with metabolic syndrome, a condition characterized by insulin resistance, central adiposity, and dyslipidemia. Prior to menopause, women have lower rates of NAFLD and associated metabolic risk factors than men; however, this gender difference disappears in older populations, implicating a protective effect of estrogen signaling.[br]To elucidate the mechanisms of estrogen action, we examined the effects of ER[alpha] knockout ([alpha]ERKO) on weight gain, glucose tolerance, and hepatic fat content in mice fed a high fat diet for 10 weeks. Relative to wild type controls, female C57Bl6 [alpha]ERKO mice (n = 6) had excess weight gain (21% increase, p[lt].05), impaired glucose tolerance (p[lt].05), and a significant increase in both micro- and macrovesicular hepatic steatosis based on blinded histology assessment (p[lt].05). Significant differences between KO and wild type mice were not observed in males. To determine if the effects of ER[alpha] are mediated by classical estrogen receptor binding to the estrogen response element (ERE), we used a murine model with a mutant ER[alpha] which does not bind DNA at ERE sites. This model, known as KI/KO, can signal through non-classical pathways but lacks direct ERE-mediated signaling in vitro and in vivo. Like [alpha]ERKO mice, female KI/KO mice exposed to 10 weeks of high fat feeding had excess weight gain (41% increase, p[lt].05), impaired glucose tolerance (p[lt].05), and increase in hepatic steatosis (p[lt].05) compared to heterozygous controls (WT/KO); again, these differences were not observed in males.[br]Taken together, these studies demonstrate (a) that signaling through ER[alpha] protects against the development of obesity, glucose intolerance, and hepatic steatosis in female mice fed a high fat diet; (b) that the effects of [alpha]ERKO on metabolic function cannot be rescued by non-classical signaling. Thus, classical ER[alpha] signaling through DNA binding is necessary for metabolic protection. Further understanding of the mechanisms by which estrogen signaling protects against liver fat accumulation and metabolic dysregulation may permit development of therapies based on selective estrogen actions in target tissues. Our findings represent a step towards development of future pharmacotherapeutics targeted to patients at high risk for type 2 diabetes and hepatic steatosis.[br][br]Sources of Research Support: NIH Fellowship Grant 1F32 DK093200-01.[br][br]Nothing to Disclose: SH-U, KH, MF, KK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1719 183 1284 SUN-230 PO08-01 Sunday 1080 2012


1078 ENDO12L_SUN-231 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Insulin Regulates Lipoprotein Lipase Activity through Angiopoietin-Like 4 in Glial Cells Sara Gry Vienberg, Andre Kleinridders, Ryo Suzuki, C Ronald Kahn Joslin Diabetes Center, Harvard Medical School, Boston, MA Lipoprotein lipase (LPL) is a key regulator of triglyceride (TG) hydrolysis into free fatty acids (FFA). Its regulation during feeding and fasting is critical for maintaining lipid homeostasis and energy supply. FFAs have been shown to suppress appetite when injected into the hypothalamus, and recent evidence has shown that TG-rich lipoproteins are sensed in the brain by an LPL-dependent mechanism, which provides lipid signals for the central regulation of body weight and energy balance. Angptl4 is a secreted protein which has been shown to regulate TG metabolism by inhibiting LPL activity. In an effort to determine how diabetes affects brain metabolism and hypothalamic function, we performed global gene expression on the hypothalamus from animal models of type 1 diabetes (streptozotocin, STZ) and type 2 diabetes/obesity (ob/ob). This revealed a significant upregulation of angiopoietin-like 4 (Angptl4) in the hypothalamus in both insulin deficient and insulin resistant diabetes compared to wildtype mice suggesting that Angptl4 is an important regulator of central lipid metabolism. Treatment of STZ mice with insulin both peripherally and directly into the cerebral ventricles restores the expression of Angptl4 to normal, suggesting that Angptl4 might also serve as an inhibitor of LPL activity and energy homeostasis in the central nervous system. In order to determine if Angptl4 regulates LPL activity centrally, we treated Angptl4 knockout (Angptl4-/-) mice and their wildtype littermates (Angptl4+/+) with STZ or vehicle and assessed LPL activity in hypothalamus and brain cortex. We did not observe any difference in LPL activity between knockout and wildtype mice in the hypothalamus or the cortex with or without STZ treatment whether fed or fasted. Furthermore, Angptl4 did not affect food intake when injected i.c.v. in wildtype mice and no difference in POMC, NPY, CART, or AgRP expression was observed between knockout mice and their controls. Interestingly, we found that Angptl4 expression is highly specific to glial cells and within these cells insulin down-regulates Angptl4 and increases LPL activity. Taken together these data suggest that Angptl4 is not involved in the regulation of brain lipid metabolism, however, it may participate in the crosstalk between neurons and glial cells.[br][br]Nothing to Disclose: SGV, AK, RS, CRK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1629 183 1285 SUN-231 PO08-01 Sunday 1081 2012


1079 ENDO12L_SUN-232 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Lack of Adipose Differentiation-Related Protein (PLIN 2) Protects Against Hepatic Lipid Accumulation but Impairs Liver Regenerative Response Against Multiple Stimuli Motoyuki Kohjima, Tsung-Huang Tsai, Lan Li, Bryan C Tackett, Sundararajah Thevananther, Benny Hung-Junn Chang, Lawrence Chan Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX Background and Aims: An early cellular response during liver regeneration after partial hepatectomy (PH) is lipid accumulation in the hepatocytes. Adipose differentiation related protein (Plin2) regulates hepatic triglyceride storage and Plin2-deficient (Plin2[minus]/[minus]) mice have significantly reduced triglyceride (TG) content in the liver. We used Plin2[minus]/[minus] mice to study the dynamics of hepatic TG content in partially hepatectomized Plin2[minus]/[minus] mice and examined whether absence of Plin2 expression modulates hepatic regeneration. Methods: We performed PH and carbon-tetrachloride toxicity on WT and Plin2[minus]/[minus] mice, and monitored the hepatic lipid profile as well as the dynamics of hepatic gene expression (by quantitative PCR) as we examined the rate of hepatic regeneration in these animals. Results and Conclusions: In wild-type mice the post-PH lipid accumulation peaked at 24h. Gene expression profiling showed that de novo lipogenesis was activated at the early phase (0-6h) whereas hepatic lipid uptake was stimulated at a later time (12-24h), allowing the lipids to be used for [beta]-oxidation. Plin2[minus]/[minus] mice showed decreased hepatic triglyceride accumulation, which was associated with impaired liver regeneration. Expression profiling showed that mRNA level of genes involved in fatty acid (FA) synthesis and lipid transfer was similar between Plin2[minus]/[minus] and WT mice, while VLDL secretion rate was elevated in the former group. We also found decreased expression of [beta]-oxidation-related enzyme genes and reduced cytosolic FA level in Plin2[minus]/[minus] mice, which may contribute to the alteration of the liver regeneration capacity in these animals. In parallel, the carbon-tetrachloride toxicity study also showed delayed liver regeneration in the Plin2 mice with down-regulation of several genes involved in lipolysis. In summary, our study supports the conclusion that normal fat storage and utilization by the liver is important for liver regeneration in liver injury and toxicity models.[br][br]Sources of Research Support: This research was supported in part by NIH grants HL 51586 (to LC), DK84495 (to BHC), and by the Diabetes [amp] Endocrinology Research Center (P30DK079638) at Baylor College of Medicine. LC was also supported by the Betty Rutherford Chair for Diabetes Research from St. Luke[apos]s Episcopal Hospital and Baylor College of Medicine, and the T.T. [amp] W.F.Chao Global Foundation.[br][br]Nothing to Disclose: MK, T-HT, LL, BCT, ST, BH-JC, LC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1746 183 1286 SUN-232 PO08-01 Sunday 1082 2012


1080 ENDO12L_SUN-233 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Dissociation of Leptin, Cholesterol, and Hepatic Scavenger Receptor SR-BI mRNA Values by High-Fat Nutritional Manipulation Aristides Lytras, Jenisa Naidoo, Krista Maxwell, Maria E Vrontakis University of Manitoba, Winnipeg, Canada; Joslin Diabetes Center, Harvard Medical School, Boston, MA; Biomedical Research Foundation, Academy of Athens, Athens, Greece; University of Manitoba, Winnipeg, Canada High fat diet (HFD) rich in saturated fatty acids (FA) induces obesity and an adverse lipid profile in humans, including hypercholesterolemia and hypertriglyceridemia. In contrast fish oil (FO), rich in polyunsaturated FA, has a lowering effect on triglyceride levels and variable effects on total cholesterol (TC) and HDL cholesterol (HDL-C) levels. In mice, hepatic HDL-C scavenger receptors (SR) have been suggested to mediate reductions in TC levels by increasing reverse cholesterol transport to the liver. Leptin has been reported to induce SR-BI expression thus contributing to adaptive responses that maintain a healthier metabolic profile in the context of adipose tissue expansion (1). To further elucidate the association of saturated and unsaturated HFD with the expression of SR-BI, adiposity and leptin serum levels, we compared 20% polyunsaturated (FO), 20% saturated (lard, L) or mixed 10% polyunsaturated - 10% saturated (FO-L) fat diets against a control chow diet (C), in 8 week-old male mice. While the composition of the HFD did not affect the dependence of leptin levels on adiposity, FO diet resulted in increased relative to C and reduced compared to FO-L or L epididymal fat mass and leptin levels. FO dramatically reduced TC levels levels compared to L, tended to decrease TC levels compared to C and FO-L, and increased hepatic SR-BI levels mRNA compared to all other diet groups. Interestingly, the mixed FO-L scheme resulted in dissociation of cholesterol, leptin and hepatic SR-BI mRNA values. Indeed, FO-L dramatically reduced TC values compared to L diet, while no differences in hepatic SR-BI mRNA levels were detected in either group compared to C. The lack of SR-BI mRNA increase in the FO-L or L diets, in spite of largely increased leptin levels compared to C, should rather be attributed to a negative effect of saturated fat on hepatic SR-BI expression. Consequently, the FO effect on cholesterol levels may be only minimally attributed to SR-BI mRNA up regulation. Indeed, while L diet appears to introduce resistance to the expected positive effects of leptin on SR-BI expression, the FO-L diet scheme maintains the beneficial impact of FO on TC compared to the L diet. Our study reveals that refined nutritional management may dissociate typical strong correlations between essential metabolic parameters providing important insights for their biological roles, as well as, for the development of nutritional modalities for therapeutic use.[br][br](1) Lund[aring]sen T, Liao W, Angelin B, Rudling M. Leptin induces the hepatic high density lipoprotein receptor scavenger receptor B type I (SR-BI) but not cholesterol 7alpha-hydroxylase (Cyp7a1) in leptin-deficient (ob/ob) mice. J Biol Chem 2003; 278:43224-8.[br][br]Sources of Research Support: Dairy Farmers of Canada (DFC) Grants Program (M. E. Vrontakis). Manitoba Health Research Council (MHRC) Post-Doctoral Fellowship (A. Lytras).[br][br]Nothing to Disclose: AL, JN, KM, MEV 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 699 183 1287 SUN-233 PO08-01 Sunday 1083 2012


1081 ENDO12L_SUN-234 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Structure, Dynamics and Cholesterol-Binding-Site Characterization of STARD6 from Solution State NMR Studies Danny Letourneau, Andree Lefebvre, Vincent Frappier, Jean-Guy LeHoux, Pierre Lavigne Facult[eacute] de M[eacute]decine et des Sciences de la Sant[eacute], Universit[eacute] de Sherbrooke, Sherbrooke, Canada; Facult[eacute] de M[eacute]decine et des Sciences de la Sant[eacute], Universit[eacute] de Sherbrooke, Sherbrooke, Canada START domain proteins belong to a superfamily of protein modules of about 210 amino acids. There are fifteen mammalian proteins that possess START domains. They are distributed into six subgroups. STARD1 and D3 of subgroup 1, and STARD4-D6 of subgroup 2 are the only ones reported to bind cholesterol. START domains adopt a helix-grip fold in which a central antiparallel [beta] sheet is surrounded by amino and carboxy terminal [alpha] helices, [alpha]1 and [alpha]4. [alpha]4 is packed over a cavity created by the curvature of the central [beta]-sheet. This cavity is proposed to be the binding site of cholesterol. From subgroups 1 and 2, STARD6 is the only START domain for which there is no three dimensional structure known. Moreover, no structure of a START domain bound to cholesterol has yet been resolved. In this context, we have determined the solution state NMR structure of the apo-form of STARD6 and characterized its molecular dynamics also using NMR methods. STARD6 adopts a classical helix-grip fold in solution. We note motions of large amplitudes located in [alpha]2 and [alpha]3 helices as well as in a [Omega] loop. These structural elements line the cavity. Motions occurring on a longer time scale are also observed in [alpha]4. Finally, amide chemical shifts displacement caused by the presence of cholesterol has allowed identifying residues that form the binding sites of STARD6. As expected, these residues line the internal cavity. These results provide unprecedented insights into the mechanism of cholesterol biding to a START domain.[br][br]Sources of Research Support: Canadian Institute of Health Research.[br][br]Nothing to Disclose: DL, AL, VF, J-GL, PL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1856 183 1288 SUN-234 PO08-01 Sunday 1084 2012


1082 ENDO12L_SUN-235 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Effect of Atorvastatin on Mitochondrial Function in Liver Cells of Rats with Diet-Induced Hypercholesterolemia and Q10 Supplement Maria Antonia Jimenez Santos, Isela Esther Juarez Rojop, Maria Teresa Espinosa Garcia, Teresa Ramon Frias, Carlos Alfonzo Tovilla Zarate, Elizabeth Carmona Diaz, Deysi Yadira Bermudez Ocana, Leova Pacheco Gil, Carlos Cuahuatemoc Diaz Zagoya Universidad Ju[aacute]rez [Aacute]utonoma de Tabasco, Comalcalco Tabasco, Mexico; Universidad Ju[aacute]rez Autonoma de Tabasco, Villahermosa Tabasco, Mexico; Universidad Nacional Autonoma de M[eacute]xico, M[eacute]xico Distrito Federal, Mexico Summary: Cholesterol is a molecule that forms part of biological membranes. Homeostasis is maintained by the processes of ingestion, absorption, synthesis, exchange and excretion. These processes are closely interconnected and any change in any of them, affects the whole process in final form, when you lose the homeostatic balance of cholesterol, hipercolesteromia originates, which is an increase of plasma cholesterol. The treatment used so far is the use of statins, which is the most widely used atorvastatin (ATV). The ATV decreases the endogenous formation of cholesterol through inhibition of the enzyme 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, however this enzyme participates in the synthesis of farnesyl pyrophosphate, precursor molecule of coenzyme Q10. The Q10 is found in several tissues, mainly in mitochondria where it is an essential component of the respiratory chain. The Q10 ensures that electrons of H (equivalent to gearboxes) are transported along the chain and therefore generating the coupling of ADP with P to synthesize ATP, molecule indispensable to generate energy.[br]Objective. To evaluate the mitochondrial function of liver cells of rats with hypercholesterolemia treated with 3 different doses of ATV and supplemented with Q10.[br]Methodology. Male Wistar rats of 180 to 200 grams, which are being grouped into 12 groups. They use an N of 6 for each group. The 10 groups were given a high cholesterol diet for 40 days. We used 3 different doses of ATV. Only 6 groups were given Q10 supplement. At the end were sacrificed, the liver was removed and homogenized, then centrifuged at 4[sup]0[/sup]C to isolate mitochondria. Mitochondrial respiration was assessed polarographically by a YSI 5300A biological oxygen monitor to 240C. 0.05.[quot][gt]They use a P[gt] 0.05.[br]Results. Respiratory control was recorded (CR) lower in the groups that received no supplement Q10. However, the groups receiving Q10 supplementation significantly increased the CR.[br]Conclusion. In the treatment of hypercholesterolemia with ATV, supplementation with Q10 improves mitochondrial respiration of hepatocytes.[br][br]Liao JK. Effects of Statins on 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase Inhibition Beyond Low-Density Lipoprotein Cholesterol. 2005. Am J Cardiol; 96: 24-33. Frederick L. Crane, PhD. Biochemical Functions of Coenzyme Q10. 2001. Journal of the American College of Nutrition; 6: 591[ndash]598. Mari M, Colell A, Morales A, Montfort C, Garc[iacute]a C, Ruiz J, C Checa F. Control of Liver Function in Health and Disease. 2010. ANTIOXIDANTS Y REDOX SIGNALING. 11: 1295-1320.[br][br]Nothing to Disclose: MAJS, IEJR, MTEG, TF, CATZ, ECD, DYBO, LPG, CCDZ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1497 183 1289 SUN-235 PO08-01 Sunday 1085 2012


1083 ENDO12L_SUN-236 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Metabolic Distribution of Cholesterol and Its Esters in the Rabbits with Hypercholesterolemia Evaluated by High-Temperature GC-MS Hyun-Jin Jung, Hong Seog Seo, Bong Chul Chung, Man Ho Choi Korea Institute of Science and Technology, Seoul, Republic of Korea; Korea University College of Medicine, Seoul, Republic of Korea Cholesteryl ester (CE) which is synthesized by the action of lecithin:cholesterol acyltransferase (LCAT) and acyl CoA:cholesterol acyltransferase (ACAT) in lipoprotein particles and cell, respectively, is major transport and storage forms of cholesterol.[br]The simultaneous profiling of CEs including cholesterol in lipid-rich biological specimens is critical to identify the cholesterol metabolism. Optimized Hybrid solid-phase extraction-precipitation (H-PPT) was accomplished to increase the extraction efficiency of cholesterol and CEs. In addition, the high temperature gas chromatography-mass spectrometry (GC-MS) technique was conducted to improve the detectability of analytes.[br]For investigating the etiology of hypercholesterolemia related metabolic disturbances, cholesterol feeding has been used to increase cholesterol levels in plasma or tissues obtained from model rabbits, which is susceptible to the development of atherosclerosis. Increased cholesterol and CE levels were observed in plasma, liver and aorta from model rabbit, whereas there was no change in kidney and muscle. The fat cholesterol levels were elevated by high-cholesterol diet feeding, but CEs were not affected.[br]It was considered that cholesterol metabolism was altered in tissue specific manner in response to cholesterol feeding.[br][br]Nothing to Disclose: H-JJ, HSS, BCC, MC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1201 183 1290 SUN-236 PO08-01 Sunday 1086 2012


1084 ENDO12L_SUN-237 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Reducing Glucocorticoid Action Improves Hyperinsulinemia but Not Insulin-Sensitive Glucose or Fatty Acid Turnover in Patients with Type 2 Diabetes with or without Fatty Liver David P Macfarlane, Peter J Raubenheimer, Mark E Bastin, Ian Marshall, Ruth Andrew, Brian R Walker University of Dundee, Dundee, UK; University of Edinburgh, Edinburgh, UK; University of Edinburgh, Edinburgh, UK Background [amp] Aims[br]Observational studies implicate glucocorticoid excess, principally due to altered steroid metabolism in target tissues, in both the insulin resistance and liver fat accumulation that accompanies type 2 diabetes. To test the contribution of glucocorticoid signalling to metabolic dysfunction we blocked cortisol secretion (with metyrapone) and action (with the GR antagonist mifepristone) simultaneously in men with type 2 diabetes [plusmn] fatty liver.[br]Methods[br]14 men with type 2 diabetes, aged 58[plusmn]2y with BMI 32[plusmn]2kg/m[sup]2[/sup], received metyrapone+mifepristone or placebo in a double-blind crossover study, administered 8h and 1h before infusions of [sup]2[/sup]H[sub]2[/sub]-glucose, [sup]2[/sup]H[sub]5[/sub]-glycerol and [sup]13[/sup]C[sub]1[/sub]-palmitate [plusmn] a low dose (10mU/m[sup]2[/sup]/min) hyperinsulinaemic clamp. Using MR spectroscopy, subjects were stratified as low (4[plusmn]1%, n=7) or high (23[plusmn]4%, n=7) liver fat. Results are mean[plusmn]SEM, compared by general linear model with repeated measures analysis.[br]Results[br]Glucocorticoid blockade increased ACTH, reduced cortisol levels, lowered fasting glucose and insulin levels and increased suppression of plasma glycerol levels by insulin (25.9[plusmn]6.2 vs 18.3[plusmn]6.5 and 50.6[plusmn]6.7 vs 39.8[plusmn]10.2%, in high and low liver fat groups respectively, p[lt]0.05). However, it had no measurable effect on glycerol turnover, glucose disposal, endogenous glucose production or fatty acid turnover at baseline or during low dose hyperinsulinaemia. Men with fatty liver were more centrally obese and insulin resistant, with increased fasting plasma glucose and insulin levels, and increased fatty acid turnover at baseline and during hyperinsulinaemia (rate of appearance of palmitate 3.8[plusmn]1.0 vs 2.5[plusmn]0.2 and 2.3[plusmn]0.3 vs 1.1[plusmn]0.2[micro]mol/kg fat free mass/min, p[lt]0.05, respectively). Fatty liver also associated with impaired suppression of fatty acid turnover (25.1[plusmn]9.1 vs 50.1[plusmn]7.2%, P[lt]0.05), glycerol turnover (14.3[plusmn]5.3 vs 34.3[plusmn]5.8%, p=0.01) and endogenous glucose production (38.6[plusmn]4.2 vs 69.1[plusmn]7.1%, p[lt]0.05) during hyperinsulinaemia. However, the effects of GC blockade did not differ between men with and without fatty liver.[br]Conclusions[br]Reducing glucocorticoid action, an approach being exemplified with 11b-HSD1 inhibitors, improves hyperinsulinaemia in patients with type 2 diabetes, enhancing suppression of glycerol concentrations during hyperinsulinaemia, but has no significant effects on glucose, glycerol, or fatty acid turnover during low dose hyperinsulinaemia and does not reverse the metabolic abnormalities accompanying liver fat accumulation.[br][br]Sources of Research Support: British Heart Foundation. Wellcome Trust.[br][br]Nothing to Disclose: DPM, PJR, MEB, IM, RA, BRW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1273 183 1291 SUN-237 PO08-01 Sunday 1087 2012


1085 ENDO12L_SUN-238 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Effects of Testosterone on Fat Metabolism in Young Healthy Males Stacy L Schmidt, Elizabeth H Kealey, Tracy J Horton, Daniel H Bessesen University of Colorado, Denver, CO There are clear differences between men and women in body fat distribution and serum lipid levels. We wondered what role testosterone (T) plays in these differences. Previous studies examining the effects of T supplementation on fat metabolism were conducted in elderly males. These studies suggested that changes in fat metabolism are likely due to changes in body composition as opposed to T per se. We sought to determine if short term alterations in T had significant effects on fat metabolism in young males. To address this question, healthy males (n=11; age: 29 [plusmn] 4.5yr; BMI: 26.3 [plusmn] 2.1) underwent two 7-day study phases in random order during which sex steroids were altered by the administration of Cetrotide[reg] (3mg initial, 0.25mg/d thereafter), Arimidex (1 mg/d), and AndroGel[sup]TM[/sup] (5.0G containing 50 mg of T/d) or placebo gel. These medications resulted in either a low hormone (LowH) condition (T = 302 [plusmn] 47 ng/dl; E2 = 16.6 [plusmn] 2.7pg/ml) or a T replete condition (RT) (T = 514.3 [plusmn] 49.5 ng/dl, E2 = 13.3 [plusmn] 2.1pg/ml). After 7 days on these medications and following the consumption of a normal breakfast and lunch, a meal test was conducted at 5pm with a meal containing 35% of daily energy needs (30% fat, 15% protein, 55% carbohydrate) and 50 mCi [1-[sup]14 [/sup]C] oleic acid. Blood samples were collected for 5h, and indirect calorimetery + [sup]14[/sup]CO[sub]2 [/sub]breath samples collected over the 24h following test meal ingestion. Subcutaneous abdominal and femoral adipose biopsies were taken 24hr after the meal test and dietary fat tracer content was determined by scintillation counting. [bold]Results[/bold]: Respiratory exchange ratio, resting energy expenditure, and [sup]14[/sup]CO[sub]2 [/sub]production were not different between LowH and RT conditions at any time point (p[gt]0.05). Area under the curve for postprandial glucose, insulin, free fatty acids, and triglycerides were also not different in LowH vs RT at any time point (p[gt] 0.05). The ratio of [sup]14[/sup]C labeled lipid in abdominal/femoral adipose tissue was significantly lower in the LowH vs RT condition (1.26 [plusmn] 0.16 and 1.94 [plusmn] 0.32; p=0.048). [bold]Conclusion[/bold]: Compared to a normal T level, 7 days of low testosterone does not affect oxidation of dietary fat, energy expenditure, or postprandial nutrient metabolism. However, it may contribute to a greater relative storage of dietary fat in femoral fat depots.[br][br]Sources of Research Support: NIDDK RO1 DK071155.[br][br]Nothing to Disclose: SLS, EHK, TJH, DHB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2110 183 1292 SUN-238 PO08-01 Sunday 1088 2012


1086 ENDO12L_SUN-239 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Vascular and Metabolic Effects of Intravenous Intralipid and Dextrose Infusion in Obese Subjects Aidar R Gosmanov, Dawn Smiley, Joselita Siquiera, Gonzalo Robalino, Limin Peng, Guillermo Umpierrez University of Tennessee Heath Science Center, Memphis, TN; Emory University School of Medicine, Atlanta, GA; Emory University, Atlanta, GA Elevated free fatty acids (FFAs) are implicated in the development of hypertension and endothelial dysfunction. We recently reported that high FFAs via intravenous (IV) infusion of polyunsaturated fatty acids (Intralipid[reg]) result in a rapid and significant rise in blood pressure (BP) and endothelial dysfunction in healthy subjects. In this study we investigated the effects of the co-administration of Intralipid with dextrose on BP, endothelial function and lipid metabolism.[br][bold]Methods: [/bold]Twelve obese, healthy subjects [ages:41[plusmn]7 yrs, BMI:32[plusmn]2 kg/m[sup]2[/sup], BP:113/65 mmHg, HOMA-IR: 2.0[plusmn]1.0] were studied on four random 8-hour admissions with IV saline, Intralipid 20%, dextrose 10% or combined Intralipid and dextrose infusion - all at 40 ml/hr rate. Brachial artery flow-mediated dilatation (FMD), BP and levels of FFAs, triglycerides, glucose, and insulin were measured at 0, 4 and 8 h of each infusion.[br][bold]Results:[/bold] Intralipid infusion alone increased systolic BP by 12[plusmn]9 mmHg (p[lt]0.001) and diastolic BP by 5[plusmn]6 mmHg (p=0.022) and decreased FMD from baseline by 3.2[plusmn]1.4% (p[lt]0.001). Dextrose infusion alone had neutral effects on BP and endothelial function. The combined infusion of Intralipid and dextrose resulted in a decrease in FMD by 2.4[plusmn]2.1% (p=0.002 vs baseline and p[gt]0.1 vs Intralipid). However, the combination of Intralipid and dextrose did not result in a significant increase in systolic (13[plusmn]22 mmHg, p=0.08) or diastolic BP (3[plusmn]3 mmHg, p[gt]0.1) from baseline. Plasma levels of FFAs increased by 1.03[plusmn]0.34 mmol/L (p=0.009) after Intralipid but FFAs were not different from baseline during saline, dextrose, or combined Intralipid and dextrose infusion. Similarly, triglyceride levels increased by 121[plusmn]97mg/dl (p=0.001) during Intralipid but rose only by 47[plusmn]51 mg/dL with combined infusion (p=0.008 vs baseline and p=0.019 vs Intralipid group). Plasma glucose and insulin significantly increased after dextrose and combined Intralipid and dextrose (both, p[lt]0.05), but did not change during saline and Intralipid alone.[br][bold]In summary,[/bold] short-term Intralipid infusion significantly increased BP; in contrast, dextrose alone or in combination with Intralipid did not increase BP. Combined Intralipid and dextrose infusion resulted in significant endothelial dysfunction. Dextrose infusion lowered Intralipid-induced elevation of FFAs and triglycerides. The metabolic and vascular effects of dextrose and lipid loads have important clinical implications in patients receiving nutrition support.[br][br]Sources of Research Support: ADA grant: 7-03-CR-35 and National Institutes of Health: MO1-RR00039.[br][br]Nothing to Disclose: ARG, DS, JS, GR, LP, GU 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 519 183 1293 SUN-239 PO08-01 Sunday 1089 2012


1087 ENDO12L_SUN-240 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Apolipoprotein B/A1 Ratio Is Associated with Cardiometabolic Risk in White and Black Adolescents Ishita Kotak, Yanbin Dong, Samip J Parikh, De-Huang Guo, Jigar Bhagatwala, Haidong Zhu Georgia Health Sciences University, Augusta, GA; Georgia Health Sciences University, Augusta, GA Background: Apolipoprotein B to apolipoprotein A1 (ApoB/ApoA1) ratio has been shown to be the strongest single lipoprotein-related atherosclerotic risk factor in adults. Atherosclerotic process begins in childhood. Better understanding of the relationship between ApoB/ApoA1 ratio and cardiometabolic risk in adolescents essential for early prevention and intervention. We aimed to evaluate the relationships of the ApoB/ApoAl ratio with cardiometabolic risk factors in a large cohort of white and black adolescents.[br]Method: Fasting lipid profiles, inflammatory markers, adipokines and blood pressure (BP) were measured in 559 adolescents aged 14-18 years (45% black, 49% females). Percent body fat (% BF) was measured by dual-energy X-ray absorptiometry. Visceral adipose tissue (VAT) and subcutaneous abdominal adipose tissue (SAAT) were determined by magnetic resonance imaging.[br]Results: Females had higher ApoB/AplAl ratio than males (0.64 vs 0.60, p=0.03). However, there was no racial difference in ApoB/ApoA1 ratio (p=0.91). The ApoB/ApoAl ratio was positively correlated with cardiometabolic risk factors, including waist circumference (r=0.30, p[lt]0.01), body mass index (r= 0.28, p[lt]0.01), total fat mass (r= 0.25, p[lt]0.01), % BF (r=0.17, p[lt]0.01), SAAT (r=0.18, p[lt]0.01), VAAT (r=0.23, p[lt]0.01), LDL (r=0.59, p[lt]0.01), total cholesterol (r=0.36, p[lt]0.01), triglyceride (r=0.26, p[lt]0.01), fasting insulin (r=0.09, p=0.04), hs-CRP (r=0.13, p[lt]0.01), leptin (r=0.17, p[lt]0.01), resistin (r=0.13, p[lt]0.01), fibrinogen (r=0.12, p[lt]0.01), ICAM (r=0.14, p[lt]0.01) and systolic BP (r=0.16, p[lt]0.01), but negatively correlated with ApoA1 (r=-0.51, p[lt]0.01) and adiponectin (r=-0.16, p[lt]0.01). Stepwise multiple regression analyses revealed that total cholesterol and waist circumference explained 12.5% and 8.5% of the variance in ApoB/ApoAl ratio, respectively.[br]Conclusion: The ApoB/ApoAl ratio is associated with fatness, inflammation, lipid profiles, adipokines and BP in black and white adolescents. Further studies are warranted to confirm the role of ApoB/ApoAl ratio in the identification and management of cardiometabolic risk in adolescents.[br][br]Nothing to Disclose: IK, YD, SJP, D-HG, JB, HZ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2151 183 1294 SUN-240 PO08-01 Sunday 1090 2012


1088 ENDO12L_SUN-241 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Fatty Acid Binding Proteins (FABP4 and FABP5), [beta]-Carotene and Insulin Resistance (IR): A 6-Month Double Blind Placebo-Controlled Study on the Effect of a Fruit and Vegetable Juice Concentrate (FVJC) in Boys Jose Atilio Canas, Ligeia Damaso, Karl J Mann, Shawn Sweeten, Prabhakaran (Babu) Balagopal Nemours Children[apos]s Clinic, Jacksonville, FL; Nemours Children[apos]s Clinic, Jacksonville, FL [bold]Background and Objectives[/bold]: Fatty acid binding proteins, FABP4 and FABP5, expressed predominantly in adipocytes and macrophages may play important roles in the maintenance of glucose and fatty acid homeostasis and are emerging as powerful risk factors for predicting cardiometabolic disease (CMD). The effect of nutritional interventions on FABP4 [amp]5 in children remains unclear.[br][bold]Method and Design[/bold]: We determined the effect of a 6-month supplementation with a fruit and vegetable juice concentrate (FVJC) plus nutritional counseling (NC) [italic]vs.[/italic] placebo plus NC on FABP4 [amp]5 in 9 normal weight and 22 overweight pre pubertal boys in a prospective randomized placebo-controlled trial. FABP4 [amp] 5, abdominal adiposity (DEXA), HOMA-IR, [beta]-carotene, retinol, retinol binding protein 4 (RBP4), C-reactive protein (CRP) and interleukin-6 (IL-6) were determined at 0, 3 and 6 months.[br][bold]Results[/bold]: FABP4 [amp] 5 were higher ([italic]P[/italic] [lt]0.01) in the obese vs. lean boys and correlated with CRP and abdominal fat mass ([italic]P[/italic] [lt]0.05 for all); only FABP4 was correlated to HOMA-IR (P[lt]0.05). In the obese cohort, FVJC reduced FABP4 (but not FABP5), HOMA-IR and abdominal fat mass ([italic]P [lt][/italic]0.05 for all) and increased [beta]-carotene ([italic]P[/italic] [lt]0.01) vs. placebo.[br][bold]Conclusion[/bold]: Taken together the data suggest that FVJC along with NC favorably impacts [beta]-carotene and FABP4, which may contribute to the attenuation of abdominal adiposity and insulin resistance in obese boys. Population wide studies are warranted to validate these findings.[br][br]Sources of Research Support: Nemours Fund for Children[apos]s Health awarded to JAC.[br][br]Disclosures: JAC: Research Materials, NSA, LLC, Collierville, TN. Nothing to Disclose: LD, KJM, SS, PB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 45 183 1295 SUN-241 PO08-01 Sunday 1091 2012


1089 ENDO12L_SUN-242 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Reversal of Nonalcoholic Fatty Liver by Thyromimetics in Not Sufficient To Restore Insulin Sensitivity Alexandro J Martagon, Jean Z Lin, Paul Webb, John Baxter, Kevin Phillips The Methodist Hospital Research Institute, Houston, TX Nonalcoholic fatty liver disease (NAFLD) is one of the most common liver diseases worldwide and almost always occurs in conjunction with insulin resistance. Despite the prevalence of NAFLD, there are currently no approved drugs for its treatment. Since thyroid hormones and their synthetic derivatives, thyromimetics, are strong effectors of lipid metabolism we investigated whether thyromimetics were capable of improving hepatic steatosis in obese mice. Treatment with either GC-1, which is capable of activating brown fat and inducing thermogenesis, or KB-2115, which does not elicit thermogenesis, led to a complete clearing of fatty liver. Surprisingly, however, only doses of GC-1 that were high enough to evoke thermogenesis led to improvements in insulin sensitivity, while lower doses of GC-1 and treatment with the liver-specific KB-2115 actually worsened insulin resistance. We conclude that while thyromimetics may be capable of clearing fatty liver, this liver-specific action may not be sufficient to restore insulin sensitivity. Instead peripheral effects of these compounds, such as the activation of brown fat and the induction of thermogenesis, may be necessary to ameliorate insulin resistance.[br][br]Sources of Research Support: NIH grant 1RC4DK090849.[br][br]Nothing to Disclose: AJM, JZL, PW, JB, KP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2280 183 1296 SUN-242 PO08-01 Sunday 1092 2012


1090 ENDO12L_SUN-243 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Association between Serum [gamma]-Glutamyltransferase and Pulmonary Dysfunction Eun Sook Kim, Sung Dae Moon, Je Ho Han, Yu-Bae Ahn Incheon St Mary[apos]s Hospital, the Catholic University of Korea, Incheon, Korea [bold]Introduction[/bold][br]Considerable evidence suggests that both elevated serum gamma-glutamyltransferase (GGT) concentrations and lung impairment are risk factors for cardiovascular disease (CVD) and mortality. We investigated whether serum GGT levels are associated with reduced lung function[italic].[/italic][br][bold]Methods[/bold][br]We conducted a cross-sectional analysis on subjects aged 30 to 80 years who visited Gangnam Severance Hospital for health checkups. Serum GGT levels with forced vital capacity (FVC) and forced expiratory volume in 1 second (FEV1) were examined in 15,076 subjects (47.2 [plusmn] 9.7 years; 53.3% of whom were men).[br][bold]Results[/bold][br]The metabolic profiles for both genders, including obesity, blood pressure, fasting glucose, total cholesterol, triglyceride, uric acid, and C-reactive protein (CRP) levels increased across GGT quartiles; however, high density lipoprotein cholesterol (HDL-C) levels decreased. In multivariable adjusted regression models, the serum GGT level was inversely associated with FEV1 and FVC % predicted but not with a FEV1/FVC % ratio in men and women. After adjusting for confounding variables including metabolic parameters and CRP, the odds ratio for restrictive lung disease was 1.22 (95% confidence interval [CI] = 1.04-1.42) per SD increase in log GGT and 1.26 (95% CI=1.03-1.54) in subjects in the highest range of GGT levels compared with those in the other quartiles. However, no association was observed between GGT levels and obstructive lung disease.[br][bold]Conclusions[/bold][br]We found an independent relationship between GGT level and impaired lung function, especially for restrictive lung patterns in the general Korean population. Further studies are required to confirm the role of GGT in predicting lung disease development, and to clarify underlying mechanisms.[br][br]Nothing to Disclose: ESK, SDM, JHH, Y-BA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 789 183 1297 SUN-243 PO08-01 Sunday 1093 2012


1091 ENDO12L_SUN-244 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Resequencing [italic]CETP[/italic], [italic]LIPC[/italic] and [italic]LIPG[/italic] Genes in Thai Subjects with Hyperalphalipoproteinemia Weerapan Khovidhunkit, Wanee Plengpanich, Sathapakorn Siriwong, Padiporn Limumpornpetch, Thiti Snabboon Chulalongkorn University, Bangkok, Thailand [italic]Objective:[/italic] Genetic factors associated with hyperalphalipoproteinemia (HALP or high levels of HDL-cholesterol) are incompletely understood. We resequenced 3 candidate genes, [italic]CETP[/italic], [italic]LIPC[/italic] and [italic]LIPG[/italic], which encode cholesteryl ester transfer protein, hepatic lipase, and endothelial lipase, respectively, in Thai subjects with hyperalphalipoproteinemia and compared with normolipidemic controls.[br][italic]Methods:[/italic] Sequence variants of [italic]CETP[/italic], [italic]LIPC[/italic] and [italic]LIPG[/italic] were identified by sequencing exons and exon-intron junctions in 64 subjects with HDL-cholesterol levels [ge]2.59 mmol/L (100 mg/dL) and compared with those of 113 normolipidemic subjects.[br][italic]Results:[/italic] We found 2 heterozygous frameshift mutations in [italic]CETP[/italic] (p.Leu262ProfsX31 and p.Val411ArgfsX6) and 2 heterozygous missense mutations in [italic]LIPC[/italic] (p.Gly141Ser and p.Val173Met). One deletion mutation and 3 point mutations in [italic]CETP[/italic] promoter were also identified. Collectively, these rare mutations were found only in the HALP group but not in the control group (7.8% [italic]vs.[/italic] 0%, [italic]P[/italic]=0.0056). One common variant of [italic]CETP[/italic] (p.Asp459Gly) was found at a higher frequency in the HALP group (23.4% [italic]vs.[/italic] 3.5%, [italic]P[/italic][lt]0.0001). Altogether, rare variants of [italic]CETP[/italic] or [italic]LIPC[/italic] and/or the common [italic]CETP[/italic] p.Asp459Gly variant were found in 29.7% of the HALP group [italic]vs.[/italic] 3.6% in the controls ([italic]P[/italic][lt]0.0001). No rare variant of [italic]LIPG[/italic] was identified.[br][italic]Conclusion:[/italic] Both common and rare DNA variants in [italic]CETP[/italic] and [italic]LIPC[/italic], but not [italic]LIPG[/italic], were more commonly found in the Thai HALP group, which could potentially contribute to high HDL-cholesterol phenotypes in our population.[br][br]Sources of Research Support: Thailand Research Fund (RSA5280008); Thailand Government Research Budget (2553-2554); Ratchadapiseksompotch Fund, Chulalongkorn University; Chulalongkorn University postdoctoral scholarship.[br][br]Nothing to Disclose: WK, WP, SS, PL, TS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 410 183 1298 SUN-244 PO08-01 Sunday 1094 2012


1092 ENDO12L_SUN-245 POSTER SESSION: Lipids [amp] Obesity Case Reports (1:30 PM-3:30 PM) Adrenalectomy as a Weight Loss Strategy in Morbid Obesity Associated with Adrenal Incidentaloma Gabriel I Uwaifo, Taniya De Silva, Kamran Rasul, Robert L Dubin, Ariane O Conrad, John Paige, William T Cefalu Louisiana State University Health Sciences Center, New Orleans, LA; Xavier University of Louisiana, New Orleans, LA; Louisiana State University Health Sciences Center, New Orleans, LA [bold]Background;[/bold] Obesity is a common major cardiometabolic risk surrogate and the prevalence of morbid obesity (MO) is increasing. Bariatric surgery (BS) remains the most consistently effective treatment for managing MO but significant preoperative weight loss is desirable and beneficial for optimal surgical outcomes. Adrenal incidentalomas (AIs) are also quite prevalent both in the general population and in obese subjects. There are no clear guidelines as to timing or necessity of unilateral adrenalectomy when AIs are found in MO subjects. We present the case of a 44 yr old African American (AA) lady with a left AI referred for weight management in whom adrenalectomy was followed with profound weight loss.[br][bold]Clinical case[/bold]; A 44 yr old AA lady was referred for weight Management including possible BS. She had a history of a left AI presumed non functional but was required to lose weight prior to consideration for adrenalectomy. She also had type 2 diabetes (DM), Obstructive sleep apnea (OSA), and hypertension (HT). Examination showed MO (initial BMI; 63.6kg/m2 and weight; 195.4kg), Acanthosis Nigricans and facial acne with hirsuitism. Abdominal CT showed a left AI 3.9x4.3cm with Hounsfeld units [lt]10. Endocrine work up showed no cathecol, aldosterone or androgen excess. Despite no biochemical features diagnostic of Cushing[apos]s syndrome (CS) she had mild hypercortisolemia suggestive of subclinical CS (elevated post overnight 1mg Dexamethasone suppression serum cortisol and elevated pm salivary cortisol despite normal 24 hr urinary cortisol). Based on the size of the AI, the patient had elective left adrenalectomy which revealed an adrenal adenoma. The patient has had progressive weight loss post surgery with her current weight [sim]7mths post surgery of 160kg, BMI; 52kg/m2. This has been associated with improvements in HT, DM and OSA. The profound weight loss post adrenalectomy makes the prospect of bariatric surgery less risky in this patient[br][bold]Conclusion;[/bold] The finding of marked weight loss following adrenalectomy for AI in this patient despite absence of overt CS suggests that elective adrenalectomy should be considered for the initial management of MO subjects with coexisting AIs especially if they demonstrate even subclinical hypercortisolemia. Adrenalectomy in such patients may be associated with profound weight loss and clinical improvement of co-morbidities that could make future bariatric surgery less risky and more successful for sustained weight management.[br][br]1.Midorikawa et al.,Clinical Endocrinology 2001; 54:797-804. 2.Rossi et al.,J Clin Endocrinol and Metab 2000;85: 1440-48. 3.Chiodini et al., J Clin Endocrinol and Metab 2010; 95:2736-45.[br][br]Nothing to Disclose: GIU, TDS, KR, RLD, AOC, JP, WTC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 63 184 1299 SUN-245 PO50-01 Sunday 1095 2012


1093 ENDO12L_SUN-246 POSTER SESSION: Lipids [amp] Obesity Case Reports (1:30 PM-3:30 PM) Successful Treatment of Hyperinsulinemic Hypoglycemia after Roux en Y Gastric Bypass with Restoration of Normal Anatomy Rodis Paparodis, Guillherme R Campos, Dawn Belt Davis University of Wisconsin School of Medicine and Public Health, Madison, WI; University of Wisconsin School of Medicine and Public Health, Madison, WI BACKGROUND: Roux en Y gastric bypass (RYGB) is highly effective for the treatment of morbid obesity. However, a complication of RYGB is the development of postprandial hyperinsulinemic hypoglycemia. Treatment for this devastating complication can be very difficult.[br]CLINICAL CASE: We report the case of a 47 year old woman with a history of RYGB, who presented with classic hypoglycemic symptoms after meals. Frequent hypoglycemia was noted on capillary glucose monitoring and with a continuous glucose monitoring system (CGMS). We confirmed post-prandial hypoglycemia with a serum glucose of 44 mg/dL 2 hours after a 75 g oral glucose load. We attempted to manage her hypoglycemia with a low carbohydrate diet, acarbose, verapamil, and the use of CGMS. Unfortunately, hypoglycemia persisted and she developed hypoglycemia unawareness leading to a motor vehicle accident.[br]We hypothesized that restoration of normal gastrointestinal anatomy would alleviate the hypoglycemia. To investigate this, we introduced a gastrostomy tube (G-tube) into the excluded stomach to allow feeding through the normal anatomy. We performed a mixed meal tolerance test (MTT) both orally and through the G-tube. With oral ingestion, there was a rapid spike in blood glucose levels to over 200 mg/dL, followed by a rapid decline to 80 mg/dL, accompanied by extremely robust first phase insulin secretion. With G-tube ingestion, there was minimal glucose excursion and normal first phase insulin secretion. Therefore, feeding through the normal anatomy prevented the large excursions in glucose and insulin after a meal.[br]Based on these results, we proceeded to reverse the RYGB. She had resolution of her hypoglycemia after reversal. We performed an oral MTT 5 months after her reversal, and saw no significant glucose excursions. In 10 months of ongoing follow up, no recurrence of hypoglycemia has been noted.[br]CONCLUSION: As far as we are aware, we report the first case of successful treatment of hyperinsulinemic hypoglycemia after RYGB with reversal of the gastric bypass and restoration of the normal anatomy. This syndrome is challenging for the physician and disabling for the patient. The underlying etiology of this syndrome remains unclear, but this experience suggests that alterations in the timing and location of food absorption are key components. Notably, our case would suggest that there are no permanent changes in pancreatic insulin production due to RYGB.[br][br]Nothing to Disclose: RP, GRC, DBD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1279 184 1300 SUN-246 PO50-01 Sunday 1096 2012


1094 ENDO12L_SUN-247 POSTER SESSION: Lipids [amp] Obesity Case Reports (1:30 PM-3:30 PM) Acquired Partial Lipodystrophy [mdash] Barraquer-Simons Syndrome: A Case Report Paula Freitas, Maria Joao Matos, Eva Lau, Davide Carvalho Centro Hospitalar S[atilde]o Jo[atilde]o, Faculty of Medicine University of Porto, Porto, Portugal [bold]Introduction:[/bold] Acquired partial lipodystrophy (APL) or Barraquer-Simons syndrome is a rare form of progressive lipodystrophy. It is characterized by gradual onset of symmetrical loss of subcutaneous fat. It usually begins in the face and progresses to the neck, upper extremities and trunk, spreading downward to lower extremities. The disease begins in childhood and is more prevalent in females. Reduced levels of complement 3 and a related circulating antibody (complement 3-nephritic factor) are frequently found, which can both suggest an autoimmune origin of the disease. Metabolic complications are less frequent than in other forms of lipodystrophy. [bold]Case report[/bold]: We report the case of a 31 years old caucasian woman, child of non-consanguineous, healthy parents, referred to our hospital for facial lipodystrophy. Her neonatal period and psychomotor development were unremarkable. She had a history of chickenpox in childhood and mumps at the age of 15. Menarche was at the age of 12. Since puberty, progressive disfiguring loss of subcutaneous adipose facial tissue was noticed and no other body regions are affected. She had no family history of lipodystrophy.Physical examination revealed severe face lipoatrophy and bilateral breast hypoplasia. Fasting glucose (70mg/dL), fasting insulin (2.7U/mL), A1C (5.2%) and oral glucose tolerance test (2 hours after glucose load 96mg/dL) were normal. Slightly increased LDL-cholesterol (139 mg/dl) and diminished HDL-cholesterol levels (54 mg/dl), were revealed. Serum C3 levels were low [18mg/dl (83 - 177mg/dl)]. Hemogram, biochemistry (including renal and liver function), IGF1, thyroid and sexual hormones were also normal. [bold]Conclusion:[/bold] The overall clinical and biochemical features of the patient led to the diagnosis of APL. We highlight the importance of giving appropriate counselling and follow up to these patients, as they are at high risk to develop nephritis, mainly membranoproliferative glomerulonephritis, and drusen. Unfortunately, as it is not possible to reverse the lost adipose tissue, surgery procedure is the only therapeutic option to improve the cosmetic disfigurement.[br][br]Nothing to Disclose: PF, MJM, EL, DC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1641 184 1301 SUN-247 PO50-01 Sunday 1097 2012


1095 ENDO12L_SUN-248 POSTER SESSION: Lipids [amp] Obesity Case Reports (1:30 PM-3:30 PM) PEG-L-Asparaginase Induced Hypertriglyceridemia Manige Konig, Rana Malek University of Maryland School of Medicine, Baltimore, MD Background: Severe hypertriglyceridemia can be caused by PEG-L-Asparaginase (PLA) in patients treated for Acute Lymphocytic Leukemia. PLA has long half-life and can be detected 15 days after IV administration. The mechanism on the lipid metabolism is manifold and different treatment options are available.(1) Interestingly, retreatment with PLA does not result in hypertriglyceridemia.[br]Clinical case: A 27 year old Hispanic female with newly diagnosed pre-B ALL presented with relapse to the hospital.[br]After relapse was confirmed by bone marrow biopsy CALGB 10403 protocol was initiated (prednisone, vincristine, idarubicin and PLA) and she received 4,150 units of IV PLA. She was noted to have elevated liver enzymes 8 days after PLA was administered.[br]A lipid panel was obtained 21 days afterwards which revealed high triglycerides level of [gt]2000 mg/dl ([lt]150 mg/dl) with total cholesterol level of 419 mg/dl ([lt] 200 mg/dl), HDL level of 25 mg/dl.[br]The PLA therapy was determined to be the etiology of the hypertriglyceridemia. She did not develop acute pancreatitis due to the elevated triglyceride levels.[br]She was initially started on low fat diet and fenofibrate 145 mg oral once per day but therapy was discontinued because of worsening liver function panel. Omega-3-fatty acid therapy 4 g per day was then initiated.[br]The patient was started on an intravenous insulin-drip with continuous dextrose 5% in water infusion to maintain a normal serum glucose level.[br]Her lipid panel improved within 2 days of the continuous Insulin with a triglyceride level of 905 mg/dl, cholesterol level of 374 mg/dl and an HDL level of 34 mg/dl.[br]The continuous insulin drip was stopped after 6 days and patient was noted to have a normal lipid panel 8 days after the Insulin drip was initiated.[br]Conclusion: Severe hypertriglyceridemia is a phenomenon which can occur after one time dose of PLA. Several treatment options are available to decrease the triglyceride levels which can be achieved by dietary interventions, oral pharmacological agents (fibrates, Omega-3 fatty acids, statins), heparin treatment, insulin treatment and plasmapharesis in cases with acute pancreatitis or hyperlipidemia associated viscosity syndrome with CNS involvement. After normalization of the lipid panel PEG-L-Asparginase can be safely given again as this phenomenon does not occur each time.(2)[br][br](1)Hoogerbrugge N, Jansen H, Hoogerbrugge PM. Transient hyperlipidemia during treatment of ALL with L-asparaginase is related to decreased lipoprotein lipase activity. Leukemia 1997 Aug;11(8):1377-9. (2) Lashkari HP, Lancaster D, Atra A, Champion MP, Taj MM. Symptomatic severe hypertriglyceridaemia with asparaginase therapy in acute lymphoblastic leukaemia (ALL) and lymphoblastic lymphoma: is rechallenging safe? Int J Hematol. 2011 Dec;94(6):571-5.[br][br]Nothing to Disclose: MK, RM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 201 184 1302 SUN-248 PO50-01 Sunday 1098 2012


1096 ENDO12L_SUN-249 POSTER SESSION: Lipids [amp] Obesity Case Reports (1:30 PM-3:30 PM) Hypobetalipoproteinemia as Unrecognized Cause of Nonalcoholic Steatohepatitis Iyad Syoufi, Hussein Yassine University of Arizona, Tucson, AZ [bold]Background[/bold]: Nonalcoholic fatty liver disease is common and usually has no complications. However, this disorder may progress to a serious liver disease. Recognition of the unusual etiologies of hepatosteatosis allows early diagnosis and intervention.[br][bold]Clinical case:[/bold] A 48-year-old healthy male was found to have elevated liver enzymes during routine labs. No history of alcohol or liver toxic medications use. Low cholesterol levels run in his father[apos]s family. Exam was unremarkable. BMI was 23%. Liver US showed fatty liver. Laboratory data revealed Alk phos of 173, ALT 120, AST 83, bilirubin 0.9, HgA1c 4.9. Viral and autoimmune hepatitis, hemochromatosis, Wilson disease, primary biliary cirrhosis, alpha 1 antitrypsin deficiency and celiac disease were ruled out. Liver biopsy confirmed steatohepatitis. Reviewing old labs illustrated an unusual lipid profile which included cholesterol of 86, TG 40, LDL 52, HDL 23, VLDL 8. VAP showed cholesterol 90, LDL 53, HDL 25, VLDL 12, TG 44, apoB 100 55 ([lt]109), HDL-2 3 ([gt]10), HDL-3 22 ([gt]30), LDL density pattern B. Vit E was 3.7 (3-16). Fasting Insulin was 2.7 (2.6-25).[br]We are reporting a case of nonalcoholic steatohepatitis secondary to familial heterozygous hypobetalipoproteinemia. An unnecessary work up would have been avoided if this diagnosis was considered earlier based on patient[apos]s lipid profile and family history. A low fat diet, periodic measurements of fat-soluble vitamins levels with supplementation of the deficient vitamins, retinal examination and genetic counseling were recommended to our patient.[br]Familial hypobetalipoproteinemia is a rare autosomal codominant disorder of apoB metabolism. Many mutations in the apoB gene have been reported, most of which encode for a truncated apoB molecule (1). Heterozygotes have cholesterol, LDL and apoB levels that are less than one-half of normal with most of the HDL in the HDL3 fraction. These patients are usually asymptomatic and diagnosed accidentally when lipid screening shows low cholesterol levels. Fatty liver has been increasingly recognized in these patients. Homozygotes are characterized by almost absent apoB and very low cholesterol levels. They present with severe fat malabsorption and manifestations of fat soluble vitamins deficiency (2).[br][bold]Conclusion[/bold]: Lipoprotein metabolism disorders should be in the differential diagnosis when evaluating patients with nonalcoholic fatty liver. Lipid profile assessment is recommended as a part of the work up for hepatosteatosis.[br][br](1) Burnett JR, Zhong S, Jiang ZG, et al. Missense mutations in APOB within the betaalpha1 domain of human APOB-100 result in impaired secretion of ApoB and ApoB-containing lipoproteins in familial hypobetalipoproteinemia. J Biol Chem. Aug 17 2007;282(33):24270-83. (2) Tarugi P, Averna M. Hypobetalipoproteinemia: genetics, biochemistry, and clinical spectrum. Adv Clin Chem. 2011;54:81-107.[br][br]Nothing to Disclose: IS, HY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 228 184 1303 SUN-249 PO50-01 Sunday 1099 2012


1097 ENDO12L_SUN-250 POSTER SESSION: Lipids [amp] Obesity Case Reports (1:30 PM-3:30 PM) Transaminitis Associated with Cholestoff[trade], an Over-the-Counter Plant Sterol Supplement Thanh Duc Hoang, Aamir Munawar Zariwala, Vinh Q Mai, Patrick W Clyde, Mohamed KM Shakir Walter Reed National Military Medical Center, Bethesda, MD Foods with plant sterols or stanols lower serum cholesterol levels. There are several over-the-counter (OTC) medications claiming to contain plant sterols. Plant sterols are generally considered to be safe without any significant side effects. We hereby report 2 patients who developed abnormal liver functions following the use of an OTC plant sterol containing medication, Cholestoff [sup]TM[/sup] (Nature Made Nutrional Products, Mission Hills, CA).[br]Case 1: A 59 y/o female with a history of primary hypothyroidism, treated with levothyroxine, was evaluated for mild dyspepsia. Physical examination revealed normal vital signs and no jaundice. Examinations of the heart and lungs were normal. The abodomen had no evidence of hepatomagaly. Since patient was taking OTC cholesterol medication, laboratory evaluation revealed normal CBC, serum albumin, and bilirubin, but elevated liver associated enzymes (LAEs) AST 429 U/L (nl 8-33), ALT 394 U/L (nl 4-36). Additional tests were negative for viral hepatitis A,B,C, hemochromotosis and Wilson[apos]s disease. Patient does not drink alcohol nor does she take any other medications. Three months prior to this visit, patient also had normal liver functions. Patient discontinued the use of Cholestoff [sup]TM[/sup] and within eight weeks, the liver functions became normal and have remained normal for the last 14 months.[br]Case 2: A 52 y/o man presented with hypercholesterolemia controlled with simvastatin 20mg daily. However, he discontinued simvastin and started self-treating with Cholestoff [sup]TM[/sup] 1 tablet thrice daily. He had mild dyspepsia and physical examination was normal. Serum LDL level was 128 mg/dL and LAEs were markedly elevated: AST 1084 U/L (14-59), ALT 988 U/L (9-72). Evaluations for viral hepatitis and hemochromatosis were negative. Eight weeks after stopping Cholestoff [sup]TM[/sup], LAEs returned normal. He was subsequently re-treated with simvastatin. Six months later, his LAEs were again elevated, AST 862 U/L and ALT 560 U/L. The patient admitted he had self-discontinued simvastatin and resumed taking Cholestoff [sup]TM[/sup]. A thorough counseling was given and since then he has had normal LAEs even on simvastatin therapy.[br]Both patients developed transaminitis while taking Cholestoff[sup]TM[/sup]. Since neither had a history of other liver disorders, it may be assumed that transaminitis was induced by Cholestoff [sup]TM[/sup]. It is not clear whether the plant sterols or other ingredients in Cholestoff [sup]TM[/sup] are responsible for inducing abnormal liver functions.[br][br]Nothing to Disclose: TDH, AMZ, VQM, PWC, MKMS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1833 184 1304 SUN-250 PO50-01 Sunday 1100 2012


1098 ENDO12L_SUN-251 POSTER SESSION: Lipids [amp] Obesity Case Reports (1:30 PM-3:30 PM) Mechanisms of Disappearing HDL-C in Multiple Myeloma Jonathan Stringer, Jessica S Lilley, Monica Agarwal, Michelle J Orsmeth, MacRae F Linton, Sergio Fazio, Fred Faas University of Arkansas for Medical Sciences, Veterans Administration, Little Rock, AR; Vanderbilt University Medical Center, Nashville, TN; Vanderbilt University Medical Center, Nashville, TN; Vanderbilt University Medical Center, Nashville, TN; Vanderbilt, Nashville, TN [bold]Introduction: [/bold]Acquired severe HDL-cholesterol (HDL-C) deficiencies are rare. True and artifactual HDL-C deficiencies have been reported (1). Interfering serum proteins should be included in the differential diagnosis of low HDL-C syndromes since these can cause lowering of HDL-C (2). We describe a case of disappearing HDL-C that led to the diagnosis of multiple myeloma.[br][bold]Case:[/bold] An 81 y/o man was seen for low HDL-C. He had multiple lipid profiles performed over many years. His initial lipid panel (1999) showed LDL-C 106 mg/dl, TG 71 mg/dl, and HDL-C 76 mg/dl. His HDL levels began falling in 2003 and by 2011 his lipid panel showed LDL-C 54 mg/dl, TG 45 mg/dl, and HDL-C [lt] 5 mg/dl. In addition, his hemoglobin and hematocrit went from 13.4 g/dl and 38.8% in 1999 to 11.8 g/dl and 34% in 2011. HDL-C was measured using a standard precipitation-based assay (1, 3). There were no medications or metabolic abnormalities to explain lowering of HDL-C, but further evaluation revealed elevated serum levels of IgG K and 30-40% plasma cells, compatible with a diagnosis of multiple myeloma.[br]The same sample assayed at the collaborating institution yielded HDL-C values of 5 mg/dl and 47 mg/dl using two different precipitation-based methods. This difference confirms an artifactual HDL-C lowering, likely due to paraproteinemia. Serum apoAI concentration was 153 mg/dl, not consistent with low HDL. Moreover, the patient[apos]s alpha migrating band on lipoprotein gel electrophoresis was similar to a control with an HDL-C of 48 mg/dl. However, artifact does not explain the significant decrease in HDL-C observed here. True deficiency can occur when paraproteins such as autoantibodies bind to HDL particles, resulting in unstable complexes with accelerated clearance (2, 4). We developed an ELISA assay to detect autoantibodies against native or modified apoAI. The patient[apos]s serum showed high titers of autoantibodies to modified apoAI but none against native apoAI. The decrease in HDL-C concentration from baseline following development of multiple myeloma suggests that modified apoAI undergoes an alternative clearance pathway leading to low HDL-C.[br][bold]Conclusion: [/bold]This case of severe HDL-C deficiency is explained in part by interference caused by plasma paraproteins, and in part due to the presence of autoantibodies against a modified form of apoAI, likely increasing its plasma clearance. Acquired HDL-C deficiency in the presence of anemia may indicate a developing gammopathy.[br][br]Nothing to Disclose: JS, JSL, MA, MJO, MFL, SF, FF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1652 184 1305 SUN-251 PO50-01 Sunday 1101 2012


1099 ENDO12L_SUN-252 POSTER SESSION: Lipids [amp] Obesity Case Reports (1:30 PM-3:30 PM) Hyperlipidemic Myeloma: Multiple Myeloma Presenting as Severe Hypertriglyceridemia: Acute Lowering with Insulin and Chronic Treatment with Anti-Myeloma Therapy Zandra Perez-Cadena, Maureen Koops, Tripathy Devjit AL Murphy VA Hospital, San Antonio, TX; University of Texas Health Science Center, San Antonio, TX [bold]Background[/bold]: Severe hypertriglyceridemia can be a life-threatening condition because of the risk of pancreatitis and hyperviscocity syndrome. We report an unusual case of severe hypertriglyceridemia associated with multiple myeloma.[br][bold]Clinical Case:[/bold] A 57 year-old man presented with nausea, vomiting, and headache for two weeks. He had a history of mildly elevated TG and was started on simvastatin and gemfibrozil a few months earlier but with poor adherence. He denied a history of pancreatitis, diabetes, HIV or hypothyroidism. He took no other medications. He denied alcohol consumption. He had a family history of diabetes and hyperlipidemia. On exam he was not obese, palmer and eruptive xanthomas were present. Admission labs included normal liver function tests, HbA[sub]1c [/sub]6.0%, TSH 1.3 uIU/ml (0.34-5.6), negative HIV test, cholesterol 329 mg/dl, TG 185 mg/dl, and HDL 22 mg/dl. Forty-eight hours later the patient developed visual blurring and a repeat lipid panel revealed: cholesterol 1061 mg/dl, TG 17,284 mg/dl, and HDL 32 mg/dl. Intravenous insulin was started and gemfibrozil and niacin was added later. TG gradually decreased to 1267 mg/dl. He was discharged on gemfibrozil and niacin. A few days later he presented with weakness and fatigue and work up revealed anemia with neutropenia. A serum protein electrophoresis showed an M-spike in the beta region that comprised 36.3% (3.6 g/dl; normal 0.6-1.1) of the total protein. Immunotyping characterized the spike as IgA kappa. Bone marrow biopsy revealed 35% plasma cells confirming the diagnosis of multiple myeloma (MM). He was treated with several cycles of bortezomib and dexamethasone resulting in remission. Even with continued poor adherence to lipid lowering medications his TG levels remained less than 1,000 mg/dl. From review of this temporal profile it is apparent that this patient[apos]s severe hypertriglyceridemia was related to MM. [bold]Conclusion:[/bold] MM usually is not associated with elevated TG but can be seen with the rare variant [ldquo][italic]hyperlipidemic myeloma[rdquo][/italic], possibly due to binding of paraproteins to lipoproteins. To our knowledge this is the highest level of TG (17,284 mg/dl) reported in such a patient. MM should be considered in the differential diagnosis of a patient with elevated triglyceride without identifiable causes. Acutely severe hypertriglyceridemia can be rapidly controlled with insulin, while the treatment of the MM can result in long-term control.[br][br]Nothing to Disclose: ZP-C, MK, TD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2205 184 1306 SUN-252 PO50-01 Sunday 1102 2012


1100 ENDO12L_SUN-253 POSTER SESSION: Lipids [amp] Obesity Case Reports (1:30 PM-3:30 PM) HDL Disappearance Syndrome Associated with Intravascular Lymphoma Andreas G Moraitis, Lita A Freeman, Alan A Remaley, Marcello Oliveira, Cliona Mary Grant, Wyndham Wilson, John Joseph DiGiovanna, Stefania Pittaluga, Smita Baid Abraham, Sarah Kaplan Browne, Freeman Fell Alexandra, Holland M Steven, Robert Shamburek National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD [bold]Background: [/bold]Intravascular large B-cell lymphoma (IVLBCL) is a rare disease entity of malignant lymphoma with poor clinical outcome due to difficulty of early and accurate diagnosis[i].[br][bold]Clinical case:[/bold] A 59-yo male with hypertension, diabetes mellitus and dyslipidemia presented with asymptomatic progressively decreasing HDL levels (46 to [lt]5 mg/dL evident 1 year prior to diagnosis). B-symptoms of the lymphoma were accompanied by clinical and biochemical features of adrenal insufficiency. Lipid values (in mg/dL) were: total cholesterol, 91; LDL, 46; TG, 199; HDL, [lt]5; and apoA-I, 30 (110-205 mg/dL). Lipoprotein electrophoresis showed no alpha-migrating HDL-C. Immunofixation and serum protein electrophoresis were normal. Imaging studies and a bone marrow biopsy were unremarkable. ACTH stimulation test was abnormal. FDG-PET scan showed increased activity of the nasal turbinates, where intravascular large B cell lymphoma was confirmed by biopsy. He received EPOCH-Rituximab chemotherapy. After the first cycle, adrenal insufficiency resolved, with normalization of cortisol response to cosyntropin stimulation. HDL levels began recovering as well (15 mg/dL). After the second cycle, HDL and apoA-I levels normalized (44 and 130 mg/dL).[br][bold]In vitro studies[/bold]: Mixing studies resulted in a 38% reduction in normal HDL, indicating that patient serum contained a factor that interfered with HDL or HDL metabolism. LCAT levels by Western analysis were not abnormally low despite strikingly low serum cholesterol esters (17% in patient vs. 75% in normal serum). The HDL subpopulation distribution in serum, investigated by two-dimensional nondenaturing gel electrophoresis[ii], was markedly abnormal, with complete absence of large HDL particles. After chemotherapy, a normal HDL subpopulation distribution was restored. Initial studies to identify the antibody (IgG and IgM) have not been successful.[br][bold]Conclusion:[/bold] We describe a case of [ldquo]HDL Disappearing Syndrome[rdquo] preceding the diagnosis of intravascular large B cell lymphoma presenting with adrenal insufficiency[iii, iv]. This is the first case in which HDL levels inversely mirror the clinical course and the response to chemotherapy of this rare form of lymphoma. Decreasing HDL may be an early marker of underlying malignancy and its resolution may reflect treatment response.[br][br][i] Nakamura S, Ponzoni M, Campo E. Intravascular large B-cell lymphoma. In: Swerdlow SH, Campo E, Harris NL, et al, eds. WHO classification of tumors of hematopoietic and lymphoid tissues. Lyon, France: IARC Press, 2008: 252[ndash]53. [ii] Asztalos BF, Tani M, Schaefer EJ. Metabolic and functional relevance of HDL subspecies. Current Opinion in Lipidology 2011, 22:176[ndash]185. [iii] Garg A, Hosfield EM, Brickner L.Disseminated intravascular lymphoma large B cell lymphoma with slowly decreasing HDL cholesterol. South Med J. 2011 Jan;104(1):53-6. [iv] Ohashi N, Aomatsu M, Mori A, Takahashi M, Shibuya T, Maruyama T, Inoue H, Takegoshi S, Yokoi T, Okuno M. Intravascular lymphoma with extremely low HDL. Intern Med. 2007;46(17):1475-7.[br][br]Sources of Research Support: This study was supported by the Eunice Shriver Kennedy National Institute of Child Health and Human Development, National Institutes of Health (NIH).[br][br]Nothing to Disclose: AGM, LAF, AAR, MO, CMG, WW, JJD, SP, SBA, SKB, FFA, HMS, RS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1009 184 1307 SUN-253 PO50-01 Sunday 1103 2012


1101 ENDO12L_SUN-254 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) The Role of Surgery in the Management of Recurrent Adrenocortical Carcinoma Timo Deutschbein, Ilknur Erdogan, Christian Jurowich, Matthias Kroiss, Marcus Quinkler, Peter Langer, Holger S Willenberg, Felix Beuschlein, Christian Fottner, Silke Klose, Anke Heidemeier, David Brix, Wiebke Fenske, Stefanie Hahner, Joachim Reibetanz, Bruno Allolio, Martin Fassnacht University Hospital Wuerzburg, Wuerzburg, Germany; University Hospital Wuerzburg, Wuerzburg, Germany; Charit[eacute] University Medicine, Berlin, Germany; University Hospital Marburg, Marburg, Germany; University Hospital Duesseldorf, Duesseldorf, Germany; Ludwig Maximilians University, Munich, Germany; University Hospital Mainz, Mainz, Germany; University Hospital Magdeburg, Magdeburg, Germany; University Hospital Wuerzburg, Wuerzburg, Germany; University Hospital Wuerzburg, Wuerzburg, Germany [bold]Objective.[/bold] The role of surgery for recurrent adrenocortical carcinomas (ACC) is not well defined. Therefore, we evaluated the outcome after surgery for tumor recurrence in patients from the German ACC Registry.[br][bold]Methods.[/bold] Only patients with first recurrence after initial R0 resection were investigated. Progression-free and overall survival (PFS, OS) after first recurrence were analyzed by Kaplan-Meier method. Cox proportional hazards regression models were used to identify prognostic factors.[br][bold]Result.[/bold] Of 154 patients with first recurrence, 101 underwent repeated surgery (R0 resection, n=78) and 99 received (additional) nonsurgical therapy. After a median interval of 6 (range 1-221) months, 144 patients (94%) experienced progressive disease. Multivariate analysis adjusted for age, sex, tumor burden, time to first recurrence (TTFR), resection status after surgery for recurrence and additional therapy indicated that only two factors were significantly associated with shorter PFS (hazard ratio for progression: TTFR [gt]12 months 1.8 [95% CI 1.2-2.5] in comparison to TTFR [le]12 months; R2 resection 3.4 [1.5-8.0] and no surgery 3.4 [1.7-7.1] in comparison to R0 resection) and OS (hazard ratio for death: TTFR [gt]12 months 3.0 [2.0-4.6] in comparison to TTFR [le]12 months; R2 resection 2.6 [1.0-6.7] and no surgery 4.2 [1.8-9.8] in comparison to R0 resection). Patients who had both TTFR [gt]12 months and R0 resection of recurrent tumors (n=22) had the best prognosis (median PFS 24 months, median OS 58 months).[br][bold]Conclusions.[/bold] The best predictors of prolonged survival after first recurrence of ACC are TTFR [gt]12 months and R0 resection. Patients with longer TTFR and tumors amenable to radical resection should therefore be operated, whereas patients with shorter TTFR or tumors not amenable to radical resection do most likely not benefit from incomplete surgery.[br][br]Nothing to Disclose: TD, IE, CJ, MK, MQ, PL, HSW, FB, CF, SK, AH, DB, WF, SH, JR , BA, MF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1187 185 1308 SUN-254 PO44-01 Sunday 1104 2012


1102 ENDO12L_SUN-255 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Preventive and Therapeutic Potential of Apigenin Against Progestin-Dependent Breast Cancer Benford Mafuvadze, Yayun Liang, Cynthia Besch-Williford, Salman Hyder University of Missouri, Columbia, MO; University of Missouri, Columbia, MO Clinical and epidemiological evidence suggests that combined estrogen (E) and progestin (P) hormone replacement therapy increases the risk of breast cancer in postmenopausal women. The biological mechanisms behind this phenomenon however, remain obscure. We showed that P increases the production of vascular endothelial growth factor (VEGF) in human breast cancer cells both in vitro and in vivo. The anti-progestin RU-486 blocks this effect, suggesting the involvement of progesterone receptors in the process (Int J Cancer, 2001, 92:469). Evidence from our laboratory using two in vivo models; human xenografts in mice (Cancer Res., 2007, 67:9929), and DMBA-induced tumors in rats (Clin Can Res, 2006, 12:4062) suggests that P stimulates growth of latent tumorigenic cells in the breast, leading to the development of palpable tumors. Since RU-486 has severe side-effects, we undertook studies aimed at identifying less toxic naturally-occurring compounds with preventive and therapeutic potential against progestin-dependent breast tumors. We recently identified apigenin, a low molecular weight flavonoid, commonly found in fruits and vegetables, which inhibited medroxyprogesterone acetate (MPA)-induced VEGF synthesis and secretion from human breast cancer cells in vitro (Menopause, 2010, 17:1055). Apigenin also suppressed the growth of MPA-dependent BT-474 human xenograft tumors in nude mice, mainly by inducing apoptosis and reducing the expression of Her2/neu and VEGF. Furthermore, in the DMBA-induced mammary cancer model, treatment with apigenin reduced MPA-linked increases in tumor incidence and multiplicity, and led to a significant delay in the appearance of the first tumor, compared with administration of MPA alone (56 vs 41 days; p[lt]0.05, LIFETEST). Immunohistochemical analysis of mammary tissue collected from treated animals revealed that apigenin reduced the levels of both VEGF and its receptor, VEGFR-2, suggesting that the flavonoid has anti-angiogenic potential. Interestingly, while apigenin decreased the overall incidence of MPA-accelerated tumors, it did not block or reverse MPA-induced mammary tissue hyperplasia, suggesting that the effects of the flavonoid are exerted at a specific stage of tumor development. Thus apigenin appears to be a compound with useful preventive and therapeutic properties against P-dependent breast cancer.[br][br]Sources of Research Support: A COR award from the College of Veterinary Medicine and by research funds from RADIL, Univ. of Missouri.[br][br]Nothing to Disclose: BM, YL, CB-W, SH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 566 185 1309 SUN-255 PO44-01 Sunday 1105 2012


1103 ENDO12L_SUN-256 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Attempt To Assess Usefulness of the Salivary Cortisol Determination as an Objective Parameter To Follow Effects of Adjuvant and Palliative Therapy with [italic]Viscum album[/italic] (Mistletoe Preparation) in Women with Breast Cancer Sabina Lewicka, Annett Staudt, Anita Glenz, Bettina Reinhard-Hennch, Heike Stammer, Jurgen J Kuehn, Judith Munzinger, Andreas Scheeweiss, Thomas Strowitzki, Cornelia von Hagens University of Heidelberg, Heidelberg, Germany; University of Heidelberg Women[apos]s Hospital, Heidelberg, Germany; University Hospital Heidelberg, Heidelberg, Germany; Protestant University of Applied Sciences, Ludwigsburg, Germany; Lukasclinic, Arlesheim, Switzerland; University of Heidelberg, Heidelberg, Germany Mistletoe extracts are claimed to improve quality of life in breast cancer patients. Although the evidence to support this view is rather weak, in German-speaking countries these preparations are frequently used in a complementary and/or alternative treatment. Nevertheless the positive effects of mistletoe therapy on quality of life could not yet be surely correlated with the tumour-related laboratory parameters. Patients with cancer are commonly depressive but often not diagnosed and adequately treated. Since cortisol is a well established biochemical parameter in the diagnostics of depression, we have undertaken a determination of cortisol, in addition to the standard questionnaires assessing quality of life and depression, prior and following mistletoe therapy in women with breast cancer. 47 Women with early (Stratum A, n = 24) and advanced (metastatic, Stratum B, n = 23) breast cancer were randomized to therapy and waiting (control) group, whereby the waiting-time for treatment was 3 months. The mistletoe preparation was self injected (s.c.) 3 times/week in ascending dosage until the final dose of 20 mg was reached after 7 weeks and maintained for the rest of the treatment period. The saliva samples for cortisol determinations were collected 3 times/day for 3 consecutive days at the beginning of the study, and then for 1 day every month. The questionnaires EORTC QLQ (C-30 and BR-23), and HADS-D were completed at the beginning and after 3 and 6 months. The statistical evaluation of the cortisol values was performed with Excel (calculations) and GraphPadPrism (determinations of significance) programmes. As expected, prior to the treatment patients with early differed from those with metastatic breast cancer regarding some subscales of the questionnaires and cortisol parameters. Metastatic cancer was associated with lower morning and higher evening cortisol values and, respectively, less steep diurnal rhythm curve. After 3 month treatment the evening cortisol decreased ca. 37% and after 6 months ca. 45% and patients with metastatic cancer tended to more pronounced decrease. Although the observed changes did not reach the statistical significance, mostly due to low number of patients and large individual variations, this preliminary study indicates that salivary cortisol could be a possible surrogate parameter for assessment of the therapeutic effects of the mistletoe extracts in patients with breast cancer.[br][br]Nothing to Disclose: SL, AS, AG, BR-H, HS, JJK, JM, AS, TS, CvH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 644 185 1310 SUN-256 PO44-01 Sunday 1106 2012


1104 ENDO12L_SUN-257 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Androgens Regulate Prostate Cancer Cell Growth and Survival Via an AMPK-PGC-1[alpha]-Mediated Metabolic Switch Jayantha B Tennakoon, Jenny J Han, Xuefeng Xia, Aijun Zhang, Olga R Ilkayeva, Christopher Newgard, Daniel E Frigo University of Houston, Houston, TX; The Methodist Hospital System, Houston, TX; Duke University Medical Center, Durham, NC Prostate cancer (PCa) is the most commonly diagnosed malignancy among men in industrialized countries, accounting for the second leading cause of cancer-related deaths. While we now know that the androgen receptor (AR) is important for progression to the deadly advanced stages of the disease, it is poorly understood what AR-regulating processes drive this pathology. Here, we demonstrate that AR regulates prostate cancer cell growth and survival via the metabolic sensor 5[apos]-AMP activated protein kinase (AMPK), a kinase that classically regulates cellular energy homeostasis. Using a combination of radiolabeled assays and emerging metabolomic approaches, we show that prostate cancer cells respond to androgen treatment by increasing not only rates of glycolysis as is commonly seen in many cancers, but also glucose and fatty acid oxidation. Importantly, this effect was dependent on androgen-mediated AMPK activity. Our results further indicate that the AMPK-mediated metabolic changes increased intracellular ATP levels through peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1[alpha])-mediated mitochondrial biogenesis, affording distinct survival and growth advantages to the prostate cancer cells. Taken together our findings converge to demonstrate that androgens co-opt the AMPK-PGC-1[alpha] signaling cascade, a known homeostatic mechanism, to increase prostate cancer cell growth and survival. The current study points to the potential utility of developing metabolic-targeted drug therapies directed towards the AMPK-PGC-1[alpha] signaling axis for the treatment of prostate cancer.[br][br]Nothing to Disclose: JBT, JJH, XX, AZ, ORI, CN, DEF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1027 185 1311 SUN-257 PO44-01 Sunday 1107 2012


1105 ENDO12L_SUN-258 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Selective Inhibition of Androgen Synthesis by Orteronel (ortl), an Investigational 17,20-Lyase Inhibitor, in Men with Metastatic Castrate-Resistant Prostate Cancer (mCRPC) David B MacLean, Walter M Stadler, Daniel Shevrin, Lowell Hart, Jingyuan Wang, Iain Webb, Robert Dreicer Millennium Pharmaceuticals, Inc, Cambridge, MA; University of Chicago Medical Center, Chicago, IL; NorthShore University Health System, Evanstion, IL; Florida Cancer Specialists, Fort Myers, FL; Cleveland Clinic, Cleveland, OH CYP17a has dual enzymatic activity: 17-hydroxylation of progesterone and 17,20-lyase conversion of pregnenolone precursors to adrenal androgens. The latter promotes survival and proliferation in CRPC. Ortl (TAK-700), a non-steroid, has shown relatively selective inhibition of 17,20 lyase (IC50 140nM) vs 17-hydroxylase (IC50 760nM) in human adrenal cell lines. [bold]Methods: [/bold]The endocrine profile of ortl was evaluated in a ph1/2, open-label, multiple-dose study in men with chemotherapy-na[iuml]ve, asymptomatic mCRPC (TAK-700_201). Pts were castrate at study entry. Ph1: ortl 100[ndash]600mg BID and 400mg BID+prednisone (pred; n=3[ndash]6/dose); ph2: four additional dose groups of 23[ndash]26 pts/dose at 300mg BID, 400 and 600mg BID+pred, and 600mg QD were studied to assess PSA response. ACTH and cortisol were measured by immunoassay and in ph2, androgens and corticosterone by LC/MS (testosterone LLQ 0.2ng/dL). ACTH (cosyntropin) stimulation tests (AST) were performed in ph1. Safety, PSA and clinical data have been previously reported (ASCO 2010, 2011). [bold]Results: [/bold]Ph1 testosterone results varied due to assay insensitivity. In Ph2, mean testosterone at baseline (8.5ng/dL) decreased to 1.2ng/dL (300mg BID), 0.9ng/dL (400mg BID+pred), 0.4ng/dL (600mg BID+pred), and 1.4ng/dL (600mg QD) after 2 cycles. DHEA-S was reduced from 46.6[micro]g/dL (baseline) to 20.7[micro]g/dL (300mg BID), 4.3[micro]g/dL (400mg BID+pred), 0.86[micro]g/dL (600mg BID+pred), and 12.1[micro]g/dL (600mg QD) after 2 cycles. In ph1, at ortl doses [le]300mg BID, ASTs were normal on days 15 and 30, but were partially blunted in 2/4 pts at 400mg BID and in 4/5 at 600mg BID. In ph 2 ([ge]300mg BID), mean ACTH was modestly increased in all non-pred groups, with corresponding increases in corticosterone. The sum of corticosterone and cortisol in non-pred groups approximated mean baseline cortisol at each dose, suggesting corticosterone is a reliable indicator of ACTH-compensated glucocorticoid function. [bold]Summary:[/bold] The dose dependent endocrine profile of ortl in men with mCRPC suggests that ortl maintains relative selectivity for 17,20-lyase over 17-hydroxylase at doses [le]300mg BID. Mathematical modeling of nonclinical and clinical data predicts that, at 300mg BID, [gt]90% of 17,20 lyase is inhibited vs 67% 17-hydroxylase. This suggests long-term administration of ortl 300mg BID may be clinically feasible without toxicities associated with activation of the ACTH-adrenal axis, which may be linked to mineralocorticoid excess.[br][br]Sources of Research Support: Millennium Pharmaceuticals, Inc.[br][br]Disclosures: DBM: Employee, Millennium Pharmaceuticals, Inc. WMS: Consultant, Takeda. JW: Employee, Millennium Pharmaceuticals, Inc. IW: Employee, Millennium Pharmaceuticals, Inc. RD: Consultant, Jansen Pharmaceuticals, Millennium Pharmaceuticals, Inc., GTX, Novartis Pharmaceuticals, EMD Serano, Sanofi-Aventis, Millennium Pharmaceuticals, Inc. Nothing to Disclose: DS, LH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1895 185 1312 SUN-258 PO44-01 Sunday 1108 2012


1106 ENDO12L_SUN-259 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) ASC-J9[reg] Inhibits Human Prostate Cancer Cell Invasion through Down-Regulation of Matrix Metallopeptidase 9 Tzu-Hua Lin, Shuyuan Yeh, Chawnshang Chang University of Rochester, Rochester, NY Prostate cancer is the most common cancer disease in men in the United States. Metastasis of prostate cancer cells is the major cause of cancer related death. It is well-known that blocking of androgen/androgen receptor (AR) signaling can efficiently suppress the androgen dependent prostate cancer growth and survival, whereas the effects on the cancer metastasis are still unclear. Previously our lab has identified the ASC-J9[reg] as a novel anti-AR compound which can suppress the androgen/AR signaling in prostate cancer cells through AR degradation mechanism. In the current study, we used human prostate cancer cell lines including LNCaP, C81, C4-2, and CWR22Rv1 to study the effect of ASC-J9[reg] on prostate cancer cell invasion. The 4 cancer cell lines were seeded in 10% CD-FBS RPMI with 10nM DHT supplied with/without 10[mu]M ASC-J9[reg] for 3 days. The pre-treated cells were then plated into matrigel coated transwell plates to perform the in vitro invasion assay. The results showed that the pre-treatment of ASC-J9[reg] can suppress the cancer cell invasion in all 4 cell lines tested in varied efficiency (LNCaP, 72%; C81, 68%, C4-2, 43%, CWR22Rv1, 81%). Further mechanism study showed that ASC-J9[reg], but not the traditional AR antagonist bicalutamide, can significantly suppress the matrix metallopeptidase 9 (MMP-9) expressions, which was correlated with cancer metastasis abilities. In brief, our data suggested that ASC-J9[reg] could suppress the prostate cancer cells (LNCaP, C81, C4-2, and CWR22Rv1) invasion through suppression of MMP-9 expression in the in vitro model. The inhibition of prostate cancer metastasis by ASC-J9[reg] in the in vivo mouse xenografted model is now under investigated.[br][br]Nothing to Disclose: T-HL, SY, CC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1809 185 1313 SUN-259 PO44-01 Sunday 1109 2012


1107 ENDO12L_SUN-260 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Four Cardiac Hormones Are Novel c-Jun-N-Terminal-Kinase 2 (JNK2) Inhibitors in Human Small-Cell Lung and Prostate Cancer Cells Meghan L Lane, Omar Santana, Chelsea D Frost, Jennifer Nguyen, Jennifer Guerrero, William P Skelton IV, Michelle Skelton, David L Vesely James A Haley VA Medical Center and University of South Florida, Tampa, FL; Wichita State University, Wichita, KS; University of Florida, Gainesville, FL [bold][italic]Background:[/italic][/bold] c-Jun-N-terminal kinase (JNK) promotes cancer development and proliferation of lung and prostate cancer cells (1,2). JNK2 kinase is the JNK kinase preferentially required for proliferation of cancer cells (1,2). Four cardiac hormones, i.e. vessel dilator, long-acting natriuretic peptide (LANP), kaliuretic peptide, and atrial natriuretic peptide (ANP) eliminate up to 86% of human small-cell lung cancers growing in athymic mice and these cancers never recur in the lifespan of the mice when treated with these cardiac hormones(3).[br][bold][italic]Materials and Methods:[/italic][/bold] The effects of these 4 cardiac hormones on human JNK2 were examined over a concentration range of 100 pM to 10 [mu]M with a specific JNK2 ELISA in human small-cell lung cancer and human prostate cancer cells.[br][bold][italic]Results:[/italic][/bold] Vessel dilator, LANP, kaliuretic peptide and ANP maximally decreased JNK2 by 89%, 88%, 77%, and 89%, respectively, each at P[lt]0.0001 in human small-cell lung cancer cells. In human prostate adenocarcinoma cells, JNK2 was maximally decreased 76%, 84%, 57%, (each at P[lt]0.0001), and 26% (P[lt]0.01) secondary to vessel dilator, LANP, kaliuretic peptide, and ANP, respectively.[br][bold][italic]Conclusion:[/italic][/bold] Four cardiac hormones are significant inhibitors (up to 89%) of JNK2 in human small-cell lung cancer cells and in human prostate adenocarcinoma cells (up to 84%) as part of their anticancer mechanism(s) of action. Thus, these new endocrine treatments that eliminate a number of different cancers [underline]in vivo[/underline] strongly inhibit JNK2 kinase activity as one of their molecular targets within cancer cells.[br][br](1) Bost F, McKay R, Bost M, Potapova O, Dean NM and Mercola D: Jun kinase-2 isoform is preferentially required for epidermal growth factor-induced proliferation of human A549 lung carcinoma. Mol. Cell. Biol. 1999; 19:1938-1949. (2) Yang YM, Bost F, Charbono W, Dean N, McKay R, Rhim JS, Depatie C and Mercola D: c-Jun NH(2)-terminal kinase mediates proliferation and tumor growth of human prostate carcinoma. Clin. Cancer Res. 2003; 9:391-401. (3) Eichelbaum EJ, Sun Y, Alli AA, Gower Jr WR and Vesely DL: Cardiac hormones and urodilatin eliminate up to 86% of human small-cell lung carcinomas in mice. Eur. J. Clin. Invest. 2008; 38:562-570.[br][br]Sources of Research Support: This work was supported in part by grants from the James and Esther King Florida Biomedical Research Program, the Florida Department of Health; and the Mama Mare Breast Cancer Foundation. The contents of this publication do not represent the views of the Department of Veterans Affairs or the United States Government.[br][br]Nothing to Disclose: MLL, OS, CDF, JN, JG, WPS, MS, DLV 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 52 185 1314 SUN-260 PO44-01 Sunday 1110 2012


1108 ENDO12L_SUN-261 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Prevalence and Impact of Hypogonadism in Cancer Patients with Muscle Wasting in a Phase IIb Enobosarm Trial Adrian S Dobs, Shontelle T Dodson, Mary Ann Johnston, Michael L Hancock, Ronald A Morton, Mitchell S Steiner Johns Hopkins University, Baltimore, MD; GTx, Inc, Memphis, TN Background[br]Hypogonadism has been associated with weight loss and poor outcomes in cancer patients. Up to 50% of males with advanced cancer are hypogonadal at presentation or during the course of treatment. Wasting in cancer patients has also been associated with a decline in physical function and performance status and has major public health significance. We conducted a Phase IIb, randomized, double blind, placebo controlled, multi-center study to evaluate the effect of enobosarm on muscle wasting and physical function in cancer patients.[br]Methods[br]Patients (n=159) were randomized to oral enobosarm (1 or 3 mg) or placebo daily for 16 wks. Patients were males [gt]45 y and postmenopausal females, had [ge]2% weight loss in the 6 mths prior to randomization, BMI [lt]35 and either NSCLC, colorectal cancer, non-Hodgkin[apos]s lymphoma, chronic lymphocytic leukemia or breast cancer. We report on the incidence and impact of hypogonadism (T[lt]300 ng/dL) in this population.[br]Results[br]Baseline testosterone levels were available for 93 of 103 men. 60% of male patients were hypogonadal at randomization. Distribution of hypogonadism was similar across cancers; however hypogonadal men were less likely to complete the study. Baseline T levels were positively correlated with weight loss (r=0.32, P=0.002,) with hypogonadal men demonstrating greater weight loss in the previous six months (median, -9.5%). Baseline physical function as measured by stair climb power was higher among eugonadal males compared to hypogonadal males (84.5 watts vs 70.6 watts; P=0.016). Enobosarm significantly improved physical function in this population regardless of baseline gonadal status (hypogonadal: 18.7%, P=0.0061; eugonadal: 13.2%, P=0.0032). The magnitude of improvement was greater in hypogonadal men.[br]Conclusions[br]Hypogonadism is common in male cancer patients and is correlated with weight loss and diminished physical function. In this randomized, placebo controlled trial, enobosarm improved physical function in both hypogonadal and eugonadal men despite poorer baseline physical function in hypogonadal patients. These data provide evidence that enobosarm may play an important role in the management of cancer related muscle wasting.[br][br]Disclosures: ASD: Investigator, Aeterna-Zentaris. STD: Employee, GTx, Inc. MAJ: Employee, GTx, Inc. MLH: Employee, GTx, Inc. RAM: Employee, GTx, Inc. MSS: Employee, GTx, Inc. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 542 185 1315 SUN-261 PO44-01 Sunday 1111 2012


1109 ENDO12L_SUN-262 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Differential mTOR and PI3K Pathway Activation in Human Bronchial Carcinoids Teresa Gagliano, Rachele Rossi, Daniela Mole, Erica Gentilin, Mariella Minoia, Federico Tagliati, Ettore degli Uberti, Maria Chiara Zatelli University of Ferrara, Section of Endocrinology, Ferrara, Italy Bronchial carcinoids (BCs) are tumors originating from endocrine cells dispersed in the respiratory epithelium. BC can be divided into typical (TBC) and atypical (ABC). TBC are in general less aggressive, smaller, and much less likely to metastasize as compared to ABC. mTOR has a central role in regulating cell growth, metabolism, and apoptosis. A differential mTOR activation in TBC vs. ABC has been previously demonstrated, suggesting that mTOR pathway profile might predict the response to mTOR-targeted therapies. Our aim is to evaluate the effects of Everolimus, an mTOR inhibitor, and NVP-BEZ 235, a dual mTOR/PI3K inhibitor, on human BC cell lines. NCI H720 (ABC) and NCI H727 (TBC) cells were treated with Everolimus or NVP-BEZ 235. Cell viability, caspase 3/7 activities and cell cycle progression were evaluated after 72 h. We found that 50 nM Everolimus inhibits cell viability by -40% in NCI H720 cells and by -10% in NCI H727 cells (P[lt]0.01); 50 nM NVP-BEZ 235 has similar effects on NCI H727, but has greater effects on NCI H720 (-50%; P[lt]0.01). Everolimus does not induce apoptosis, but promotes an accumulation in G1/G0 (+10% in NCI H727 and +25% in NCI H720 cells vs. ct). NVP-BEZ 235 does not promote apoptosis in NCI H727, but induces this process in NCI H720 (+50%), and causes G1/G0 accumulation in both NCI H727 (+10%) and NCIH720 (+35%), with a parallel decrease in cyclin D1 levels, in GSK3[beta] phosphorilation and in ERK1/2 activation. In addition, mTOR expression is greater, at both protein and mRNA levels, in NCI H720 cells as compared to NCI H727 (1.5 fold).[br]These data demonstrate that mTOR and PI3K pathway are differentially activated in human BCs cell lines, depending on stage (TBC vs. ABC) and mTOR expression levels. These data support previous findings showing that Everolimus reduces cell viability of selected human primary coltures of BC. Moreover, NVP-BEZ 235 could have a role in the medical treatment of ABCs.[br][br]Nothing to Disclose: TG, RR, DM, EG, MM, FT, EdU, MCZ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 565 185 1316 SUN-262 PO44-01 Sunday 1112 2012


1110 ENDO12L_SUN-263 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Glucagon-Like Peptide 1 Receptor (GLP-1R) Imaging for the Preoperative Localization of Benign Insulinomas: Experience in 30 Patients Emanuel Christ, Flavio Forrer, Damian Wild, Beat Gloor, Thomas Clerici, Michael Braendle, Helmut Maecke, Peter J Ell, Martyn E Caplin, Jean Claude Reubi University Hospital of Bern, Inselspital, Bern, Switzerland; University Hospital of Basel, Basel, Switzerland; University Hospital of Freiburg, Freiburg, Germany; University Hospital of Bern, Inselspital, Bern, Switzerland; Kantonsspital St Gallen, St Gallen, Switzerland; Kantonsspital St Gallen, St Gallen, Switzerland; University Hospital of Basel, Basel, Switzerland; University College Hospital, London, UK; Royal Free Hospital, London, UK; University of Bern, Bern, Switzerland [bold][italic]Background:[/italic][/bold][br]Although biochemical diagnosis of endogenous hyperinsulinemic hypoglycemia is straightforward, surgical removal of an insulinoma is hampered by difficulties to localize it using conventional radiological procedures (endosonography, MRI, CT-imaging techniques).[br]In vitro data suggest that human insulinoma cells exhibit a high density of GLP-1R. [sup]111[/sup]In-exendin-4 is a [sup]111[/sup]In labeled GLP-1R agonist that binds with high affinity to GLP-1R and may be helpful in localizing benign insulinomas.[br][bold][italic]Aim:[/italic][/bold][br]To localize benign insulinomas using [sup]111[/sup]In-exendin-4 in patients with proven endogenous hyperinsulinemic hypoglycemia but no or only one suspicious lesion on conventional imaging.[br][italic]Material and Methods:[/italic][br][sup]111[/sup]In-exendin-4 was administered intravenously at a dose of ([sim]90 MBq; 30 ug peptide) over 5 minutes to 30 patients (18 females, 12 males, age 52.6 [plusmn] 14.8 years, mean [plusmn] SD; range 28-75years). Whole body planar images and SPECT/CT images (Single Photon Emission Computed Tomography) of the abdomen were performed at 0.5 h, 4 h, 23 h, 96h up to 168h post injection. Diagnosis was confirmed by histology after surgical removal. In vitro determination of GLP-1R was performed using autoradiography.[br][italic]Results:[/italic][br]Conventional imaging (MRI, CT, endosonography) was positive in 17 patients. [sup]111[/sup]In-exendin-4 SPECT/CT detected 23 true positive benign insulinomas and five additional positive lesions (1 malignant insulinoma; 2 islets hyperplasia; 2 uncharacterized lesions). True negative tests were detected in 2 patients (1 malignant insulinoma; 1 islets hyperplasia). There was no false negative result. The sensitivity of the method was 100%, the positive predictive value 82%. In 17 patients with insulinoma an excellent correlation between GLP-1R SPECT/CT and in vitro determination of GLP-1R could be documented.[br][italic]Conclusion:[/italic][br]These data confirm results from a pilot study (1) and suggest that in vivo GLP-1R imaging defines a new non-invasive diagnostic approach to successfully localize small insulinomas.[br][br]1) Christ ER and al., JCEM, 2009;94: 4398.[br][br]Sources of Research Support: SNF (PASMP3-123269) Oncosuisse (No. OCS-02259-08) King[apos]s College London and UCL Comprehensive Cancer Imaging Centre CR-UK [amp] EPSRC, in association with the MRC and DoH (England).[br][br]Nothing to Disclose: EC, FF, DW, BG, TC, MB, HM, PJE, MEC, JCR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 218 185 1317 SUN-263 PO44-01 Sunday 1113 2012


1111 ENDO12L_SUN-264 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) How Clinical Presentation of Insulinoma Is Changing Marco Toaiari, Maria Vittoria Davi, Letizia Boninsegna, Massimo Falconi, Giuseppe Francia Internal Medicine D, Verona, Italy; Ospedale Sacro Cuore di Negrar, Verona, Italy [bold]Introduction:[/bold] In 2009 the Endocrine Society[apos]s clinical practice guidelines for management of hypoglycemic disorders, confirming previous suggestions, have stated that the new cut-off of insulin for the diagnosis of insulinoma should be lowed to 3[micro]U/mL in presence of hypoglycemia (serum glucose [le] 55 mg/dL). Moreover hypoglycemia in post-prandial status has been reported in insulinoma, alone or associated with fasting hypoglycemia. Finally preexisting diabetes mellitus is increasingly recognized in these patients.[br][bold]Aim[/bold]: to evaluate clinical features and diagnostic criteria in a monocentric series of insulinomas including sporadic and multiple endocrine neoplasia type 1 (MEN-1) patients. In addition anamnestic glucose metabolic profile was investigated.[br][bold]Patients and methods:[/bold] clinical and biochemical data regarding 33 patients including 27 sporadic and 6 MEN1 were retrospectively analyzed.[br][bold]Results:[/bold] the 72 hour fasting test was positive in all cases. However in one case it was initially negative since hypoglycemia did not occur, but became positive after 2 year follow-up. Three patients showed a nadir insulin level [gt] 3 [mu]U/ml but [lt] 6 [mu]U/ml. In the remaining 30 cases lowest insulin level was [gt] 6 [mu]U/ml. At presentation 27 patients (82%) reported only fasting symptoms, 3 (9%) only post-prandial symptoms and 3 (9%) both. Four cases (12%) had been previously affected by type 2 diabetes mellitus and 4 cases by impaired fasting glucose[br]C[bold]onclusion: [/bold]the new cut-off of insulin has further increased the sensitivity of 72h fasting test for the diagnosis of insulinoma from 87 to 100%. However the absence of hypoglycemia during test does not rule out in definitive manner the diagnosis and the test should be repeated in any high suspicion case. Patients with insulinoma can also present with only post-prandial hypoglycemia. Surprisingly previous glucose metabolism alterations are not infrequent and require further evaluation in wider series.[br][br]Nothing to Disclose: MT, MVD, LB, MF, GF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1593 185 1318 SUN-264 PO44-01 Sunday 1114 2012


1112 ENDO12L_SUN-265 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Estimated Penetrance of Main Tumors in Multiple Endocrine Neoplasia Type 1 during Second Decade of Life: An Attempt in Optimizing Periodic Screening of Adolescent MEN1 Carriers Tatiana Denck, Rodrigo Toledo, Sergio Toledo, Delmar Lourenco, Jr University of S[atilde]o Paulo School of Medicine, S[atilde]o Paulo, Brazil Context: Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant inherited disorder with high susceptibility to developing endocrine tumors in pituitary (PIT) and parathyroids glands (HPT) and endocrine cells from duodenum and pancreatic islets (PET). Genetic status of index cases and at-risk familial members were possible after MEN1 gene discovery. In order to reach an early diagnosis, the Consensus on MEN1 (2001) established periodic hormonal/radiological exams in MEN1 carriers with beginning to each tumor type based on younger case age reported so far. Recently, non-functioning PETs were diagnosed in younger ages (2nd decade) than that defined by Consensus (after 20 y-old). The penetrance of different tumors at the 2nd decade is not well known.[br]Objectives: To recognize the penetrance and prevalence of main MEN1 tumors during 2nd decades reaching a more appropriate screening. [br]Casuistic: One hundred and fourteen MEN1 cases presenting germline MEN1 mutations were studied.[br]Results: The prevalences and penetrances of HPT, PET and PIT in 27 cases with MEN1 diagnosis younger than 21 y-old were respectively 95.2%, 70%, 70% and 74.1%, 53.8%, 56%. The penetrances of HPT, insulinoma, gastrinoma, non-functioning PET, PIT, prolactinoma and non-functioning PIT in 114 MEN1 cases were respectively 27.2, 2.6, 0, 47.4, 15.7, 13 and 1.9%. Half of cases diagnosed during 2nd decade were asymptomatic and predominant symptoms were related to prolactinoma (70%), insulinoma (20%) and HPT (10%). During follow-up, 26.6% of young cases with asymptomatic HPT presented urolithiasis before 21 years. Prolactinoma was the more prevalent pituitary tumor (77.8%) and 44.5% were macroadenoma. Non-functioning PITs are less frequent (22%) presenting as incipient microadenomas. Non-functioning PETs are frequent into 2[ordf] decade (60%) and relevant clinically once 55% of the cases presented criteria to surgical treatment. [br]Conclusions: Our data indicate to periodic vigilance of symptoms related to prolactinomas, HPT and insulinomas with laboratory screening during 2[ordf] decade, including MRI if hyperprolactinemia is present. Active investigation of non-functioning PITs and PETs with MRI should be conducted between 15-20 and 10-15 y-old respectively. Long-term prospective studies will contribute to better definition of the selection, beginning and periodicity of exams to screening and early diagnosis of relevant clinically tumors in MEN1 carriers adolescents.[br][br]Sources of Research Support: Sources of Research Support: This study was supported by Sao Paulo State Research Foundation (FAPESP) Grants #2008/58552. FLC is the recipient of a National Council for Scientific and Technological Development (CNPq) fellowship (142603/2010-4), RAT is the recipient of a Sao Paulo State Research Foundation postdoctoral fellowship (11942/2009) and SPAT is the recipient of a National Council for Scientific and Technological Development senior fellowship and a National Council for Scientific and Technological Development grant (401990/2010-9).[br][br]Nothing to Disclose: TD, RT, ST, DL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2089 185 1319 SUN-265 PO44-01 Sunday 1115 2012


1113 ENDO12L_SUN-266 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Effect of Everolimus on the Pharmacokinetics of Octreotide LAR in Patients with Advanced Neuroendocrine Tumors: A RADIANT-2 Analysis Marianne E Pavel, Kai Grosch, Wing Cheung, Jens Hasskarl, Carlos Becerra Charit[eacute] Universit[auml]tsmedizin Berlin, Berlin, Germany; Novartis Pharma AG, Basel, Switzerland; Novartis Pharmaceuticals Corporation, East Hanover, NJ; Novartis Pharmaceuticals AG, Basel, Switzerland; Texas Oncology-Baylor Charles A Sammons Cancer Center, Dallas, TX [bold]Background:[/bold] In RADIANT-2 (NCT00412061), everolimus plus octreotide long-acting repeatable (LAR) exhibited a clinically meaningful 5.1-month improvement in progression-free survival (PFS) versus placebo plus octreotide LAR in patients with advanced neuroendocrine tumors (NET) and a history of carcinoid syndrome (16.4 months vs 11.3 months; hazard ratio [HR], 0.77; 95% CI, 0.59-1.00; one-sided log-rank test, [italic]P[/italic]=0.026; prespecified boundary, [italic]P[/italic][le]0.0246). A secondary endpoint was to characterize the effects of the coadministration of everolimus on the pharmacokinetics of octreotide LAR.[br][bold]Methods: [/bold]429 patients with advanced NET received everolimus (10 mg/day) plus octreotide LAR (30 mg IM q28d; n=216) or placebo plus octreotide LAR (n=213). The effect of everolimus on octreotide trough concentrations (C[sub]min[/sub]) was assessed by a linear mixed-effects model that included dose and treatment as fixed effects and subject as a random effect.[br][bold]Results:[/bold] Coadministration of everolimus plus octreotide LAR increased octreotide C[sub]min[/sub] with a geometric mean ratio (everolimus/placebo) of 1.47 (90% CI, 1.32-1.64). Adjusted geometric mean octreotide C[sub]min[/sub] at week 4 was 5.19 ng/mL (n=141; everolimus group) and 3.53 ng/mL (n=148; placebo group), respectively. Cox regression determined that the overall impact of octreotide logC[sub]min[/sub] on PFS prolongation was not statistically significant in either the everolimus ([italic]P[/italic]=0.17) or the placebo ([italic]P[/italic]=0.13) arm. Importantly, the risk for progression was consistently reduced when everolimus C[sub]min[/sub] was increased 2-fold, regardless of octreotide exposure (6 ng/mL octreotide coadministration: HR, 0.74; 95% CI, 0.46-1.18) (4 ng/mL octreotide coadministration: HR, 0.54; 95% CI, 0.32-0.92). This suggests that the antiproliferative activity of everolimus was independent of exposure to octreotide.[br][bold]Conclusions:[/bold] Coadministration of everolimus plus octreotide LAR increased octreotide C[sub]min[/sub]. A model-based analysis of PFS determined that the antiproliferative effect of everolimus observed in RADIANT-2 does not appear to be influenced by octreotide LAR exposure. Increase in everolimus C[sub]min[/sub] reduced the risk of progression.[br][br]Sources of Research Support: Novartis Pharmaceuticals Corporation.[br][br]Disclosures: MEP: Consultant, Ipsen, Novartis Pharmaceuticals, Pfizer, Inc.; Speaker, Ipsen, Novartis Pharmaceuticals, Pfizer, Inc. KG: Employee, Novartis Pharmaceuticals. WC: Employee, Novartis Pharmaceuticals. JH: Employee, Novartis Pharmaceuticals. Nothing to Disclose: CB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1766 185 1320 SUN-266 PO44-01 Sunday 1116 2012


1114 ENDO12L_SUN-267 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Upregulation of sst[sub]2[/sub] by the Demethylating Agent 5-aza-2-Deoxycytidine Augments Uptake of Radiolabeled Somatostatin Analogue in Human sst[sub]2[/sub] Expressing Cell Lines Marije J Veenstra, Peter M van Koetsveld, William E Farrell, Fadime Dogan, A Marlijn Waaijers, Diana M Sprij-Mooij, Steven WJ Lamberts, Giovanni Vitale, Leo J Hofland Erasmus Medical Centre, Rotterdam, Netherlands; Keele University, Stoke on Trent, UK; University of Milan, Milan, Italy; IRCCS Istituto Auxologico Italiano, Milan, Italy Introduction[br]In neuroendocrine tumors the somatostatin receptor type 2 (sst[sub]2[/sub]) is a target for therapy with somatostatin analogues (SSA). However, there is a high variability in tumoral receptor expression and response to SSA treatment between patients. In cell lines it has been shown that promoter CpG methylation influences sst[sub]2[/sub] expression. We hypothesize that upregulation of sst[sub]2[/sub] expression by the demethylating drug 5-aza-2-deoxycytidine (AZA) leads to augmented uptake of the radiolabeled SSA [[sup]125[/sup]I-Tyr[sup]3[/sup]]octreotide by tumor cells.[br]Materials and Methods[br]8 Human sst[sub]2[/sub] expressing tumor cell lines of various origin were exposed to increasing doses of AZA to determine the effect on cell proliferation. The effect of AZA (IC[sub]50[/sub]) on sst[sub]2[/sub] expression was evaluated in all cell lines. In BON (Carcinoid), DMS-79 (Small Cell Lung Ca) and PANC-1 (Pancreatic Ca) the effect of AZA (IC[sub]20[/sub] and IC[sub]50[/sub]) on the uptake of [[sup]125[/sup]I-Tyr[sup]3[/sup]]octreotide was determined by internalization experiments. By pyrosequencing, CpG island methylation around the transcription start site (TSS) of the sst[sub]2[/sub] gene promoter region was also determined in the cell lines.[br]Results[br]AZA significantly increased sst[sub]2[/sub] expression (range 1.4-4.1 fold) in 6 of 8 cell lines. In BON and PANC-1, AZA induced a 2.5-3 fold increased uptake of [[sup]125[/sup]I-Tyr[sup]3[/sup]]octreotide and a significant dose-dependent increase of sst[sub]2[/sub] mRNA expression. In DMS-79 there was no significant increase of sst[sub]2[/sub] mRNA expression, however, there was a significant 50% increase in internalization of [[sup]125[/sup]I-Tyr[sup]3[/sup]]octreotide. Methylation percentages at the CpG positions surrounding the TSS correlated inversely with sst[sub]2[/sub] mRNA expression.[br]Discussion[br]Measurement of internalized [[sup]125[/sup]I-Tyr[sup]3[/sup]]octreotide by BON, PANC-1 and DMS-79 revealed an augmented dose dependent uptake after treatment with AZA. In BON and PANC-1 the increased uptake is likely due to upregulation of sst[sub]2[/sub] expression. In DMS-79 there is no significant upregulation of sst[sub]2[/sub] expression under the influence of AZA, so an indirect mechanism is suggested. Pyrosequencing results in DMS-79 cells reveal very low CpG methylation levels that can explain the absence of an effect of AZA on sst[sub]2[/sub] mRNA. A relationship between CpG methylation around the TSS and mRNA expression of sst[sub]2[/sub] is reinforced by the correlation data. Enhanced uptake of (radiolabeled) SSA may increase the efficacy of targeted therapy making AZA a potential interesting addition to the therapy.[br][br]Nothing to Disclose: MJV, PMvK, WEF, FD, AMW, DMS-M, SWJL, GV, LJH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1912 185 1321 SUN-267 PO44-01 Sunday 1117 2012


1115 ENDO12L_SUN-268 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Keloids: A New Phenotype Associated with Wild-Type GISTs and No Mutations in the Succinate Dehydrogenase Enzyme Evgenia Gourgari, Kim Su Young, Eva Szarek, Margarita Raygada, Constantine Stratakis National Institute of Child Health and Human Development, Bethesda, MD; National Institute of Child Health and Human Development, Bethesda, MD Gastrointestinal Stromal Tumors (GISTs) are rare mesenchymal neoplasms of the GI tract originating from the pacemaker cells known as the interstitial cells of Cajal. The majority of GISTs are caused by activating somatic mutations in KIT or PDGFRA; cases that do not have these mutations are termed wild-type (WT). WT GISTs can occur sporadically, or as part of Carney Stratakis syndrome (along with paragangliomas-PGLs), Carney Triad, Neurofibromatosis type-1 (NF-1),Von-Hippel Lindau disease (VHLD) and, very rarely with other endocrine and non-endocrine tumors. Recently, inherited germline mutations in subunits of the mitochondrial succinate dehydrogenase (SDH) complex B, C, D, A, and AF2 (SDHx) have been implicated in the pathogenesis of wild-type GIST. We have previously reported that up to 29% of patients with WT GIST tumors harbor germline SDH-gene mutations, in the absence of co-existing paraganglioma; a more recent study (Janeway et al. PNAS2011) put this number at 12%. The National Institutes of Health has established a clinic for WT GIST; this is a bi-annual collaborative work between clinicians, researchers, support groups and patients. We describe here the characterization of a clinical phenotype observed in the majority of patients with WT GIST focusing on facial features and skin findings. We propose that this phenotype may help effectively discriminate between GIST subgroups. We analyzed the data of 53 patients with wild type GIST; 43 had normal germline SDHx sequence and 10 had abnormal germline SDH sequence. Of those ten patients, 4 had pathogenic mutation for SDHB, 3 had an SDHB sequence variant, 2 had a mutation in SDHC and 1 had a mutation in SDHD. We compared measurement of inner canthal distance, outer canthal distance and the presence or absence of keloids between those that had normal sequence of SDH versus those that had an abnormal sequence. We found a statistically significant difference in the presence of keloids in patients without SDHx mutations or variants versus those with abnormal sequence (p=0.0386 by Fisher[apos]s exact test). There were no significant differences in facial measurements. We conclude that alternate genetic events may predispose to the formation of keloids in patients with GISTs and no SDHx mutations or variants. We speculate that this has to do with activation of novel molecular pathways in skin fibroblasts that may also be responsible for GIST pathogenesis.[br][br]Nothing to Disclose: EG, KSY, ES, MR, CS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2154 185 1322 SUN-268 PO44-01 Sunday 1118 2012


1116 ENDO12L_SUN-269 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Expert Panel Consensus Statements on the Medical Treatment of Unresectable Pancreatic Neuroendocrine Tumors George Fisher, Jonathan R Strosberg, Al B Benson, Jennifer L Malin, Lowell B Anthony, Bulent Arslan, John F Gibbs, Edward Greeno, Renuka Iyer, Michelle K Kim, William Maples, Philip A Philip, Edward Wolin, Dasha Cherepanov, Michael S Broder Stanford University Medical Center, Stanford, CA; University of South Florida College of Medicine, Tampa, FL; Robert H Lurie Comprehensive Cancer Center of Northwestern University, Chicago, IL; University of California, Los Angeles, CA; University of Kentucky Medical Center, Lexington, KY; Rush University Medical Center, Chicago, IL; State University of New York at Buffalo, Roswell Park Cancer Institute, Buffalo, NY; University of Minnesota, Minneapolis, MN; State University of New York at Buffalo, Roswell Park Cancer Institute, Buffalo, NY; Mount Sinai Medical Center, New York, NY; Mission Health System, Asheville, NC; Wayne State University School of Medicine, Karmanos Cancer Institute, Detroit, MI; Samuel Oschin Cancer Center, Cedars-Sinai Medical Center, Los Angeles, CA; Partnership for Health Analytic Research, LLC, Beverly Hills, CA Background: Neuroendocrine tumors (NETs) of the pancreas (PNETs), a major subtype of gastrointestinal NETs, are rare neoplasms that lack some specificity in current treatment guidelines. We describe a physician expert panel consensus on medical treatment of well-differentiated (grade 1-2) unresectable PNETs.[br]Methods: PNET treatment appropriateness ratings were collected using the RAND/UCLA Delphi process: recruited physician experts (e.g., by specialty, geography, practice), reviewed treatment literature, and collected 2 rounds of ratings (before and after a face-to-face meeting) from the experts. Experts and the moderator were blinded to the funding source. Patient scenarios (rated on a 1-9 scale indicating appropriateness of interventions for a given scenario) were labeled as appropriate, inappropriate, or uncertain. No appropriateness rating was assigned to a scenario in presence of disagreement: [gt]2 ratings from 1-3 and [gt]2 from 7-9 range.[br]Results: Ten panelists (mean age: 50.4 years) from the northeast, midwest, south, and west census regions convened for a 1 day meeting. Specialties represented were medical and surgical oncology, interventional radiology, and gastroenterology. Panelists had practiced for a mean of 15.5 years (range: 6-33). Among 202 rated scenarios, disagreement decreased from 13.2% (26 scenarios) before the meeting to 1% (2) after. In the 2[sup]nd[/sup] round, 46.5% (94 scenarios) were rated inappropriate, 21.8% (44) were uncertain, and 30.7% (62) were appropriate. Consensus statements from the scenarios included: 1) it is appropriate to use somatostatin analogs (SA) as 1[sup]st [/sup]line therapy in patients with hormonally functional tumors, 2) it is appropriate to use everolimus, sunitinib, or cytotoxic chemotherapy therapy as 1[sup]st [/sup]line therapy in patients with symptomatic or progressive tumors, and 3) beyond 1[sup]st[/sup] line, these same agents can be used as can octreotide LAR (in patients with uncontrolled secretory symptoms) in doses up to 60 mg every 4 weeks or up to 40 mg every 3 or 4 weeks.[br]Conclusions: We systematically obtained appropriateness ratings for a variety of medical therapies in PNETs from a group of physician experts. The Delphi process allowed the experts to reliably quantify complex qualitative data in order to arrive at consensus on the appropriateness of medical therapies for the treatment of PNETs.[br][br]Disclosures: JRS: Consultant, Novartis Pharmaceuticals, Genentech, Inc.; Researcher, Novartis Pharmaceuticals, Amgen, Pfizer, Inc. AIBB: Consultant, Bayer, Inc.; Researcher, Bayer, Inc. LBA: Consultant, Pfizer, Inc., Novartis Pharmaceuticals, Pfizer, Inc., Lilly USA, LLC; Consultant, Amgen, Novartis Pharmaceuticals; Researcher, ImClone Systems, Novartis Pharmaceuticals. RI: Researcher, Pfizer, Inc. PAP: Consultant, Sanofi-Aventis, Sanofi-Aventis. EW: Consultant, Novartis Pharmaceuticals, Pfizer, Inc., Ipsen; Researcher, Novartis Pharmaceuticals, Pfizer, Inc., Ipsen. DC: Researcher, Novartis Pharmaceuticals. MSB: Principal Investigator, Novartis Pharmaceuticals. Nothing to Disclose: GF, JLM, BA, JFG, EG, MKK, WM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1726 185 1323 SUN-269 PO44-01 Sunday 1119 2012


1117 ENDO12L_SUN-270 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Prognostic Value of WHO Grading vs. AJCC Staging in Pancreatic Neuroendocrine Tumors Emilie Morin, Stefano Serra, Sonia Cheng, Sylvia Asa, Shereen Ezzat University of Toronto, Toronto, Canada; University of Toronto, Toronto, Canada Introduction. Pancreatic Neuroendocrine Tumors (pNETs) are the second most common malignancy of the pancreas with an estimated incidence of 0.5 per 100,000 and a complex spectrum of clinical behaviors. Here we describe the clinical and pathological characteristics associated with long-term prognosis.[br]Methods: We retrospectively reviewed 119 patients with pNETs treated in a tertiary referral center with a median follow-up of 38 months (range 1-360). Pathologic characteristics including size, grade, and immunohistochemistry (IHC) profile were analyzed. Primary clinical outcomes were stability or progression of the tumor and disease-free survival (DFS).[br]Results: The mean age at presentation was 52 years (SD 13.49), 56% were female, 9% were associated with MEN 1, 3% with VHL. The clinical presentation was incidental in 39%, followed by endocrine hypersecretion syndromes (24%) and abdominal pain (17%). Mean tumor diameter was 3.3 cm (SD 2.92). While 45% were considered non-functional, hormone functionality was confirmed by IHC in 55%: 30 insulinomas (27%), 17 glucagonomas (15%), 8 VIPomas (7%), 2 gastrinomas (2%) and 1 somatostatinoma (1%). According to the WHO 2010 classification, 55% were low grade (G1), 42% intermediate grade (G2), and 3% high grade (G3). Extra-pancreatic extension was noted in 22%.[br]Disease progression (23.5% of cases) occurred more frequently in higher AJCC stages (I: 6.8%, IIA 8.7%, IIB: 26.7%, IIIB: 28.6%, IV: 94.4%, p [lt] 0.0001) and in higher WHO grades (G1: 10%, G2: 26.3%, G3: 100%, p = 0.002). We found stable disease in 66.4% of cases and WHO grade was significantly correlated with this behaviour (G1: 82% remained stable, G2: 57.9%, G3: 0, p = 0.002).[br]DFS was shorter in non-functioning tumors than functioning tumors (78 vs. 145 months, p=0.03), in VIPomas than glucagonomas (1 vs. 240 months, p=0.01), in G3 tumors than G2 tumors (21 vs. 144 months, p[lt]0.011), and with liver involvement (20 vs. 173 months, p[lt] 0.005), and positive lymph nodes (33 vs. 208 months, p[lt]0.005). DFS was also significantly associated with AJCC stage. Age, gender, number of primary tumors and heredity were not associated with prognosis.[br]Conclusion: AJCC stage and WHO grade emerged as markers of pNET disease progression. Stage, grade, function, presence of LNM and/or liver metastases were independently associated with poor prognosis for DFS.[br][br]Nothing to Disclose: EM, SS, SC, SA, SE 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 318 185 1324 SUN-270 PO44-01 Sunday 1120 2012


1118 ENDO12L_SUN-271 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Correlation of AJCC/UICC TNM Staging and WHO Grading with Prognosis of Intestinal NETs Paula Araujo, Ozgur Mete, Sonia Cheng, Sylvia L Asa, Shereen Ezzat University of Toronto, Toronto, Canada; University of Toronto, Toronto, Canada The increasing incidence and heterogenous behavior of neuroendocrine tumors (NETs) of the gut pose a clinicopathological challenge. Our goal was to compare the prognostic value of the 2010 WHO (World Health Organization) grading and the AJCC/UICC TNM, 7[sup]TH[/sup] Edition (American Joint Committee on Cancer) staging systems of NETs arising in the intestine.[br]Methods: Retrospective data from 91 patients with intestinal NETs were collected including clinical presentation, imaging studies, serum biomarkers, treatment, pathology characteristics and survival. Response Evaluation Criteria In Solid Tumors (RECIST) was used to define recurrence or progression. All cases were graded using the WHO 2010 system and and staged using the AJCC/UICC TNM system.[br]Results: 50.5% of patients were male with a mean age of 56 years (range 17-84). Mean overall survival (OS) was 139 months (95% CI, 114-163 months). 93% underwent surgery and 44% were treated with octreotide LAR. Carcinoid syndrome, present in 34% of cases, was an independent predictor of metastases to the liver (p[lt]0.0001) and of death of disease (p=0.016). Stages I, II, III and IV included 8, 8, 37 and 38 patients respectively. Lower stages had significantly higher estimated disease free survival (DFS) (p[lt]0.0001), but without impact on OS. WHO grading was available for 70 patients; the distribution was 40, 29 and 1 for grades 1 (G1), 2 (G2) and 3 (G3), respectively. Estimated disease free survival (DFS) was similar for G1 and G2 groups, whereas a higher OS was inversely proportional to the grade (G1: 40 months, G2: 36, G3: 15, p = 0. 031).[br]A higher OS was observed among patients who used octreotide LAR post-operatively (Stage III; mean survival 84.1 vs. 48.1, p= 0.035) and (Stage IV; 76.1 vs. 34.2 months, p= 0.005). WHO grading also predicted differences in OS among octreotide-treated patients (G1: mean survival 68.6 mths vs. 40.5, p = 0.021; G2: 67.5 vs. 35.3, p = 0.046). In contrast, comparison of DFS according to both grading and staging systems by survival analysis showed no significant differences in octreotide-treated patients.[br]Conclusion: Both WHO grading and AJCC/UICC TNM staging systems have some limitations in predicting OS. For patients who undergo octreotide LAR treatment, WHO grading stratifies OS patients with G1 and G2 and AJCC staging remains a better predictor of DFS and OS. Our findings underscore the complementarity of clinicopathologic staging and grading in providing estimates of NET disease outcomes.[br][br]Nothing to Disclose: PA, OM, SC, SLA, SE 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 319 185 1325 SUN-271 PO44-01 Sunday 1121 2012


1119 ENDO12L_SUN-272 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) The Pheochromocytoma Susceptibility Gene [italic]TMEM127[/italic] Regulates the Early Endosome Yuejuan Qin, Yilun Deng, Patricia L Dahia UTHSCSA-Univ Texas Health Science Center at San Antonio, San Antonio, TX We recently identified [italic]TMEM127[/italic] as a tumor suppressor gene involved in susceptibility to pheochromocytomas[sup]1,2[/sup]. In vitro inactivation and mutation of [italic]TMEM127[/italic] lead to increased phosphorylation of mTOR targets. Furthermore, TMEM127 colocalizes with multiple components of the endomembrane system, suggesting that it may participate in the regulation of early events of protein trafficking. To further define the degree of interaction between TMEM127 and endosomal components, we investigated the early endosomal marker Rab5. Expression of a constitutively active Rab5 mutant (Rab5Q79L,GTP-bound Rab5) in TMEM127-overexpressing HeLa cells increased the fraction of TMEM127 that colocalizes to the early endosome, while defective Rab5 mutant S34N (GDP-bound Rab5) led to loss of TMEM127 association with early endosomal markers, indicating that TMEM127 membrane association is dependent on the GTP status of Rab5. In further support of the relevance of Rab5 for TMEM127 function, colocalization with early endosomal markers was abrogated in TMEM127 mutants identified in pheochromocytoma patients.[br]Rab5 activation due to expression of the Rab5Q79L mutant has been shown to inhibit mTOR signaling by preventing conversion of early to late endosomes and thereby restricting access of mTOR to its late endosomal activators. We found that TMEM127 cotransfection with Rab5Q79L enhanced the effect of this Rab5 mutant on mTOR downregulation. In contrast, siRNA-mediated depletion of [italic]TMEM127[/italic] markedly inhibited the ability of Rab5Q79L mutant to generate hybrid early/late endosomal vesicles, suggesting that loss of TMEM127 facilitates endocytic traffic progression from the early endosome. Furthermore, [italic]TMEM127 [/italic]knockdown also led to reduced expression and redistribution of endogenous Rab5 signals toward the cell periphery, implying that TMEM127 is required for Rab5 localization. Taken together with our previous findings, these data suggest a mechanism for TMEM127 modulation of mTOR whereby TMEM127 limits early-to-late endosomal conversion and might thus restrict mTOR access to its site of activation at the late endosome/lysosomal domain. These findings support a role for TMEM127 in endomembrane trafficking and signal regulation.[br][br]1. Qin Y et al. Germline mutations in TMEM127 confer susceptibility to pheochromocytoma. Nat Genet, 2010. 42(3): p. 229-33. 2. Yao L et al. Spectrum and prevalence of FP/TMEM127 gene mutations in pheochromocytomas and paragangliomas. JAMA, 2010. 304(23): p. 2611-9.[br][br]Sources of Research Support: This work was supported by grants from the Voelcker Fund,the Cancer Prevention and Research Institute of Texas (CPRIT), the Alex[apos]s Lemonade Stand Foundation, the Concern Foundation.[br][br]Nothing to Disclose: YQ, YD, PLD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1768 185 1326 SUN-272 PO44-01 Sunday 1122 2012


1120 ENDO12L_SUN-273 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Characterization of Pheochromocytomas in MEN 2 with [italic]RET[/italic] Codon 918 and 634 Mutations Sonali N Thosani, Naifa Busaidy, Maria Cabanillas, Gilbert Cote, Elizabeth Grubbs, Mohammed Habra, Mimi Hu, Nancy Perrier, Montserrat Ayala-Ramirez, Theresa Rich, Steven G Waguespack, Camilo Jimenez, Anita Ying MDACC, Houston, TX Background:[br]Pheochromocytomas (PHEO) occur in up to 50% of patients with MEN 2 but have not been extensively characterized. It is unknown whether the presence of PHEO affects the medullary thyroid cancer (MTC) phenotype in MEN2. The aim of this study is to present our institutional experience with MEN2-associated PHEO and to assess whether or not PHEO impacted survival from MTC.[br]Patients and Methods:[br]Through retrospective review of the institutional database of pheochromocytoma patients, we identified PHEOS with germ line [italic]RET [/italic]mutations. The following variables were evaluated: (1) clinical data: age at diagnosis of MTC and pheochromocytoma, [italic]RET [/italic]mutation, diagnosis through screening, family history of pheochromocytomas and medullary thyroid cancer, and (2) median overall survival. We chose patients with [italic]RET[/italic] codon 634 and 918 mutations for survival comparison analysis to a control population of 74 patients (56 (634) and 18(918)) without PHEO and with [italic]RET [/italic]positive MTC.[br]Results:[br]We identified 71 patients with MTC + PHEO and [italic]RET[/italic] codon mutations: 59(634), 7(918), 3(618), 1(666), and 1 (883). The median age at diagnosis of pheochromocytoma was 32 years and of MTC was 22 years. 58% of patients presented with MTC as initial manifestation of MEN 2, and 35% with concomitant MTC and PHEO. Only one patient had an apparently sporadic PHEO. None of the patients had metastases from PHEO. 59% had a family history of pheochromocytomas. 67% of patients were diagnosed through biochemical screening for catecholamines. The median size of pheochromocytoma at diagnosis was 3.5 cm and the median size of MTC at diagnosis was 1.5 cm. 72% of patients had bilateral pheochromocytomas, which were synchronous in 81% of patients and metachronous in 19%.77% of patients had open surgery for removal of pheochromocytoma and 64% had cortical sparing surgery. Survival analyses comparing MTC + PHEO patients with [italic]RET[/italic] 634 and 918 to a control group with MTC only but carriers of [italic]RET [/italic]918 and 634 showed no significant difference. The median survival time for patients with MTC + PHEO and MTC only was 502 months and 417 months, respectively ([italic]p[/italic] [gt]0.05).[br]Conclusions:Pheochromocytomas in the setting of MEN 2 are usually bilateral and most frequently associated with [italic]RET [/italic]mutation 634. Most PHEOS are currently detected due to biochemical and genetic screening. Our study did not show that the presence of pheochromocytoma conveyed a worse survival compared with MEN 2 patients without pheochromocytomas.[br][br]Nothing to Disclose: SNT, NB, MC, GC, EG, MH, MH, NP, MA-R, TR, SGW, CJ, AY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2285 185 1327 SUN-273 PO44-01 Sunday 1123 2012


1121 ENDO12L_SUN-274 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Simultaneous Body PET/MRI for SDHx-Related Metastatic Paraganglioma: Initial NIH Results Elise M Blanchet, Victoria Martucci, Corina Millo, Clara C Chen, David A Bluemke, Karel Pacak National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; University Hospital of Angers, Angers, France PET imaging has a major role in paragangliomas, for (re)-staging as well as for tumor characterization. However, an MRI is often necessary due to its higher soft tissue contrast. Moreover, limiting radiation exposure is preferable, especially for young patients. The aim of this study was to test the feasibility of simultaneous body PET/MRI on patients with SDHx-related metastatic paragangliomas and to assess its diagnostic value versus PET/CT using our new hybrid PET/MRI system.[br]Three initial adult patients (2 SDHB, 1 SDHD) with metastatic paragangliomas were evaluated with an 18F-FDG body PET/CT, immediately followed by a body PET/MRI, without the injection of any additional radiotracer. The PET and basic 3 Tesla MRI data were acquired simultaneously on a Biograph mMR (Siemens Healthcare) in 30 minutes. Then, a complete contrast-enhanced MRI was performed on the same machine. This standard MRI as well as a contrast-enhanced CT and clinical and imaging follow-up, served as the standard of reference for the accuracy assessment of the PET/MRI.[br]The datasets showed excellent image quality and good correlation between the SUVs obtained with the PET/MRI and those obtained with the PET/CT (R=0.93). A total of 32 lesions were evaluated. The lesions were located in the liver (4), bone (19), parasympathetic and sympathetic ganglia stations (7), and lung (2). Two retroperitoneal lesions were detected on both PET/CT and PET/MRI, but much more precisely defined on PET/MRI due to the intrinsic value of co-registered non contrast-enhanced MR images. The same observation was noted for all other liver lesions. However, one lung lesion (5 mm) was missed by PET/MRI.[br]In conclusion, our initial results suggest that simultaneous body PET/MRI is a very efficient imaging modality in the evaluation of SDHx-related metastatic paragangliomas. It could be particularly beneficial for the pediatric population (given the substantial radiation dose reduction), for precise lesion definition in organs where MRI can be of added value to CT (liver, abdomen, head and neck) and for following locoregional liver therapies (radiofrequency ablation, embolization). In the future, with the development of new PET radiotracers and the use of PET for treatment response assessment, as well as with the possibility of performing functional MRI sequences, we can expect substantially improved paraganglioma characterization, resulting in improved personalized treatment.[br][br](1) Boss A et al, Eur Radiol 2011; 1439:1446. (2) Delso G et al, J Nucl Med 2011; 1914:1922. (3) Schwenzer NF et al, Abdom. Imaging. 2012; 20:28. (4) Havekes B et al, Clin. Endocrinol. 2010; 137:145.[br][br]Sources of Research Support: Intramural Research Program of the National Institute of Child Health and Human Development, National Institutes of Health.[br][br]Nothing to Disclose: EMB, VM, CM, CCC, DAB, KP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2152 185 1328 SUN-274 PO44-01 Sunday 1124 2012


1122 ENDO12L_SUN-275 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Succinate Dehydrogenase Gene Mutations in Urinary Bladder Paraganglioma Zarina Lorenzo, Tamara Prodanov, Alex Ling, Karen T Adams, Karel Pacak National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD [bold]Background:[/bold] Paraganglioma (PGL) of the bladder (UBPGL) is a rare disease that accounts for less that 0.05% of all bladder neoplasms and less than 10% of PGLs. Urinary PGL was first described by Zimmerman in 1953 and usually presents with a triad of sustained hypertension, hematuria and postmicturation syncope. Several studies have shown that mutations in the genes encoding succinate dehydrogenase (SDH) subunits B, C, D cause extra-adrenal PGLs associated with familial PGL syndromes PGL4, PGL3, PGL1 respectively. In this study we investigated PGLs affecting the urinary bladder and we described their clinical characteristics, biochemical phenotype and genetics.[br][bold]Methods:[/bold] Charts of patients diagnosed with pheochromocytoma and PGL were reviewed and those with pathologically confirmed PGL of the urinary bladder seen at the National Institutes of Health in Bethesda, Maryland, USA from 2000 to 2011 were included. Genetic testing for mutations and deletion in SDHB, C, and D was performed. Plasma and urinary catecholamines and metanephrines were measured and imaging scans (CT scans, MRI, FDG or [sup]123[/sup]I MIBG scans) were reviewed.[br][bold]Results: [/bold]A total number of 11 patients were diagnosed with UBPGL. Of the eleven, 5 were males and 6 were females. The mean age was 35.27 [plusmn]15.52 (range 8-57 years). Eight out of 11 (72%) patients were hypertensive. Ten out of 11 patients presented with signs and symptoms of catecholamine excess (palpitations, flushing, sweating, pallor and headaches) at the time of diagnosis. Most of the patients had noradrenergic phenotype and 3 had both noradrenergic and dopaminergic phenotype. Only 2 (18%) presented with metastasis upon initial diagnosis. All were diagnosed within a year of a clinical presentation except for 2 patient who were diagnosed 9 and 10 years after the onset of symptoms. Overall 91% of patients developed metastatic disease and 2 (18%) had recurrent disease. In only 8 (72%) out of 11 patients, CT scans were positive for UBPGL. Only 4 had MRI scanning which all showed the urinary bladder mass. Majority of the patients, 7 (64%) had an SDHB related UBPGL.[br][bold]Conclusion: [/bold]Urinary bladder paraganglioma is commonly associated with SDHB mutations. Patients who present with UBPGL should have SDHB testing as a first line of genetic testing.[br][br]Nothing to Disclose: ZL, TP, AL, KTA, KP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2153 185 1329 SUN-275 PO44-01 Sunday 1125 2012


1123 ENDO12L_SUN-276 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Exposure to Beta-Blockers and ACEI/ARB Is Associated with Improved Survival in Patients with Advanced Colon Cancer Diana R Engineer, Basil O Burney, Jose M Garcia Michael E Debakey Veterans Affairs Medical Center, Houston, TX; Baylor College of Medicine, Houston, TX; St Luke[apos]s Episcopal Hospital, Houston, TX; Baylor College of Medicine, Houston, TX Background: Advanced colon cancer is associated with decreased survival. Recent in-vitro studies suggest that angiotensin and beta-adrenergic blockade may decrease colon cancer progression. Moreover, this may have a beneficial effect on the host by preventing the increased REE or inhibiting lipolysis (beta blockers, BB) and muscle proteolysis (angiotensin converting enzyme inhibitors/angiotensin receptor blockers, ACEI/ARB).[br]Objective: Determine whether exposure to BB, ACEI or ARB is associated with decreased mortality, weight loss, or number of hospitalizations in patients with advanced colorectal cancer.[br]Methods: This is a retrospective chart review of patients diagnosed with advanced colorectal cancer at our institution between January 2000 and July 2009. Patients with hypertension and diabetes were included, but patients with other conditions that may benefit from angiotensin blockade or from BB (i.e. NYHA Class III or IV CHF, end-stage COPD, hyperthyroidism) were excluded. Data collected included survival, hospitalization, gender, cancer progression, cancer treatment, body weight (6-12 mo pre-diagnosis, baseline and every 6 mo post-diagnosis), albumin, creatinine, medications, and stage.[br]Results: From the 425 new diagnoses of stage III-IV colon cancer reviewed, 262 patients met the inclusion criteria. Patients exposed to ACEI/ARB, BB or both were most likely to have diabetes, HTN and to be stage III. Hence, stage was included as a covariate in all analyses. There was no difference in age, gender, chemotherapy regimen or radiation between groups. Exposure to ACEI/ARB+BB combination was associated with decreased mortality compared to unexposed individuals even after adjusting for age and stage (median survival 1341 v 695 days respectively, Cox regression p[lt]0.03). Body weight changes between 6 and 18 months after diagnosis also predicted survival after adjusting for stage and exposure. BB+ACEI/ARB exposure was associated with more weight gain compared to unexposed subjects (p[lt]0.02), and it predicted weight loss even after adjusting for stage. A tendency towards increased cancer progression (p=0.085) in unexposed patients was also noted. Number of hospitalizations was similar between groups.[br]Conclusion: Exposure to a combination of BB and ACEI/ARB is associated with weight gain and increased survival in patients with advanced colorectal cancer. Future studies to determine the causality of this association are warranted.[br][br]Disclosures: JMG: Investigator, Aeterna Zentaris Inc. Nothing to Disclose: DRE, BOB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 283 185 1330 SUN-276 PO44-01 Sunday 1126 2012


1124 ENDO12L_SUN-277 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) B-Lymphocyte Stimulator: A New Potentially Useful Serological Marker in the Follow-Up of Patients with Neuroendocrine Tumors Franco Grimaldi, Martina Fabris, Elio Tonutti, Daniela Visentini, Stefano Pizzolitto, Giovanna De Maglio, Miriam Isola, Francesco Curcio, Fabio Vescini, Sveva Macrini University Hospital SM Misericordia, Udine, Italy; University Hospital SM Misericordia, Udine, Italy; University Hospital SM Misericordia, Udine, Italy; University Hospital SM Misericordia, Udine, Italy; University of Udine, Udine, Italy; University of Udine, Udine, Italy Aim of the study. To confirm B-Lymphocyte Stimulator (BLyS) as a new possible prognostic marker in the follow-up of patients with neuroendocrine tumors (NET).[br]Methods. One hundred-one (n. 101) consecutive patients with NET and 56 sex-matched controls enrolled in the study. Patients were classified in 2 subgroups according to clinical course: evidence of persistent but stable disease or in remission (n = 59) and patients with evidence of recurrent disease (n = 42). BLyS and Chromogranin A (CgA) serum levels were analyzed by ELISA at baseline (n. 101) and at multiple time points in the follow-up (2 points n. 45; 3 points n. 12), registering disease behavior.[br]Results. BLyS levels were up-regulated in NET patients compared to control subjects (1129[plusmn]489 pg/ml vs 655[plusmn]158 pg/ml; p [lt]0.0001) and correlated with tumor differentiation (1042[plusmn]371 pg/ml in gastroenteric G1 - typical lung carcinoid vs 1277[plusmn]590 pg/ml in gastroenteric G2 [ndash] atypical lung carcinoid; p=0.019; unpaired t-test) and disease behavior (912[plusmn]248 pg/ml in pts with stable disease/remission versus 1433[plusmn]578 pg/ml in pts with recurrent disease; p [lt]0.0001). In the follow-up (overall 11.8[plusmn]8.6 months, range 3-36) BLyS levels remained in the normal range ([lt]939 pg/ml) in patients with stable disease (n. 14, from 877[plusmn]275 pg/ml to 844[plusmn]209 pg/ml; p=ns; fig. 1), decreased in pts going from active disease to stable disease/remission (n. 10, from 1402[plusmn]262 pg/ml to 1105[plusmn]269 pg/ml; p=0.002; fig. 2), but increased in patients with further disease progression (n. 14, from 1665[plusmn]842 pg/ml to 2054[plusmn]1221 pg/ml; p=ns; fig. 3) or relapsing (n. 7, from 1020[plusmn]184 pg/ml to 1307[plusmn]179 pg/ml; p=0.031; fig. 4). Longer follow-ups in a preliminary limited number of cases confirmed such results (Figures 5 to 7). In contrast, CgA levels showed conflicting changes. Metastatic patients displayed higher BLyS levels than non-metastatic ones (1282[plusmn]657 pg/ml vs 1085[plusmn]385 pg/ml; p=0.07).[br]Conclusion. Elevated BLyS levels characterized more aggressive NET cases. BLyS appears as a new potential prognostic marker in the follow-up.[br][br]Nothing to Disclose: FG, MF, ET, DV, SP, GDM, MI, FC, FV, SM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 268 185 1331 SUN-277 PO44-01 Sunday 1127 2012


1125 ENDO12L_SUN-278 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Humanin, a Mitochondria-Derived Peptide, Protects Against Doxorubicin Cardiotoxicity Griselda Alvarez, Changhan Lee, Pinchas Cohen University of California, Los Angeles, CA Doxorubicin (DXR), a widely used antitumor anthracycline antibiotic, causes severe cardiomyopathy and congestive heart failure, limiting its long-term clinical usefulness. Increased oxidative stress is a major factor implicated in DXR-induced cardiotoxicity. We hypothesized that [bold]Humanin (HN)[/bold], a small peptide encoded within the mitochondria, may enhance stress resistance against DXR-induced cardiotoxicity. Humanin is a novel 24 amino acid peptide encoded within the 16S ribosomal RNA region of the mitochondrial DNA. It was cloned independently by three separate groups and described to be a potent cytoprotective peptide. Humanin has been shown to have a wide range of protective effects including [italic]in vitro[/italic] studies of amyloid beta-mediated neuronal cytotoxicity where it has been suggested that oxidative stress plays a key role by triggering mitochondrial oxidative damage. Other protective effects of HN have been shown [italic]in vivo [/italic]in Alzheimer and diabetes models. Humanin injections also protect against the development of atherosclerosis in APOE-KO mice and against ischemia reperfusion injury in the heart. In this study we pretreated mice with HN or scrambled peptide control (i.p.) for 48 hours prior to DXR treatment (10 mg/kg) and show protection from cardiotoxicity. Plasma HN levels were at least 50-fold elevated compared to endogenous levels at the time of sacrifice, as determined by an in-house ELISA. Using Seahorse, we measured both basal and maximum cardiac oxygen consumption rate (OCR) to estimate the degree of DXR-induced mitochondrial damage. HN pretreatment sustained both basal OCR and maximum respiration capacity of cardiac mitochondria 20% higher than controls. Mice did not show behavioral abnormalities, changes in body weight or blood glucose levels following HN treatment, confirming its safe use as shown in previous studies. In addition, we showed that DXR infusion doubled circulating HN levels, possibly representing an acute stress response. These studies suggest that HN may enhance protection from mitochondrial toxins.[br][br]Nothing to Disclose: GA, CL, PC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2365 185 1332 SUN-278 PO44-01 Sunday 1128 2012


1126 ENDO12L_SUN-279 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Acute Metabolic Changes Following the Levothyroxine Withdrawal in Patients with Differentiated Thyroid Carcinoma Kyung Ae Lee, Heung Yong Jin, Sunhee Kim, Jae Moon Kim, Hong Sun Baek, Tae Sun Park Chonbuk National University Medical School, Jeonju, Republic of Korea; Chonbuk National University-Chonbuk National University Hospital, Jeonju, Republic of Korea Aim: After total thyroidectomy, differentiated thyroid carcinoma (DTC) patients have to undergo levothyroxine (LT4) withdrawal for measuring serum thyroglobulin and 131I whole-body scan to evaluate residual or recurrent disease. LT4 therapy is usually withdrawn for 4 weeks (during the first 2 weeks patients receive treatment with LT3 instead of LT4, and the last 2 weeks stop all medication). There are a few reported studies looking into the effects of LT4 withdrawal in DTC patients in terms of biochemical parameters. We evaluated the effects of iatrogenic hypothyroidism in patients with DTC on lipid profile, renal function, liver function, electrolyte.[br]Method: Serum levels of total cholesterol, low density lipoprotein (LDL), triglycerides, creatinine, aspartate aminotransferase (AST) and alanine aminotransferase (ALT), electrolyte were determined in 60 DTC patients during levothyroxine treatment and 4 weeks after LT4 withdrawal.[br]Results: After T4 withdrawal, mean TSH value and mean free T4 value were 87.9 mIU/L and 1.7 pmol/L, respectively. Statistically significant increases were found in total cholesterol, LDL and triglycerides with LT4 withdrawal (4.5 mmol/L vs 6.4 mmol/L, 2.5 mmol/L vs 3.9 mmol/L, 1.4 mmol/L vs 2.0 mmol/L, respectively). In addition, AST and ALT concentrations increased (21 U/L vs 50 U/L, 12 U/L vs 58 U/L, respectively). Creatinine levels increased significantly and sodium decreased on stopping LT4 treatment (61.5 umol/L vs 83.2 umol/L, 142 mmol/L vs 139 mmol/L, respectively).[br]Conclusions: Acute hypothyroidism induced by LT4 withdrawal in DTC patients during monitoring for remnant or metastatic disease, seriously affects multiple organs and systems, especially older patients or those with other chronic diseases. This study demonstrate that short term hypothyroidism associated with a significant increase in risk factors for atherosclerosis, decreased renal function and hepatic function, electrolyte imbalance. Therefore, in the follow-up of DTC patients, L-T4 withdrawal procedure should be restricted to cases where the cost/benefit ratio is favorable. Alternative methods, such as the use of recombinant human TSH or image modality (neck US, CT, PET-CT), should be considered.[br][br]Nothing to Disclose: KAL, HYJ, SK, JMK, HSB, TSP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 198 185 1333 SUN-279 PO44-01 Sunday 1129 2012


1127 ENDO12L_SUN-280 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) A Novel Multitarget Tyrosine Kinase Inhibitor with Antiangiogenic Properties, Active against Anaplastic Thyroid Cancer [italic]In Vitro[/italic] and [italic]In Vivo[/italic]: CLM3 Alessandro Antonelli, Guido Bocci, Concettina La Motta, Silvia Martina Ferrari, Poupak Fallahi, Andrea Di Domenicantonio, Ilaria Ruffilli, Gabriele Materazzi, Romano Danesi, Federico Da Settimo, Paolo Miccoli University of Pisa, School of Medicine, Pisa, Italy; University of Pisa, School of Medicine, Pisa, Italy; University of Pisa, School of Medicine, Pisa, Italy; University of Pisa, School of Medicine, Pisa, Italy Introduction. We have studied the antitumor activity of a new [ldquo]pyrazolo[3,4-d]pyrimidine[rdquo] compound (CLM3) proposed for a multiple signal transduction inhibition (including the RET tyrosine kinase, EGFR, VEGFR) and with antiangiogenic activity, in primary anaplastic thyroid cancer (ATC) cells in vitro and in vivo.[br]Methods. CLM3 was tested: a) in the human cell line 8305C (undifferentiated thyroid cancer) at 0.001-100 [micro]M; b) in ATC cells at the concentrations of 5, 10, 30, 50 [micro]M; c) in an ATC-cell line (AF), derived from a primary ATC culture, in CD nu/nu mice.[br]Results. CLM3 significantly inhibited proliferation of 8305C cells, inducing also apoptosis. A significant reduction of proliferation with CLM3 in ATC cells (P [lt] 0.01, ANOVA) was shown. CLM3 increased the percentage of apoptotic ATC cells dose-dependently (P [lt] 0.001, ANOVA) and inhibited migration (P [lt] 0.01, Newman-Keuls test) and invasion (P [lt] 0.001, Newman-Keuls test). AF-cell line was injected sc in CD nu/nu mice and tumor masses became detectable 25 days after. CLM3 (40 mg/kg.die) inhibited significantly tumor growth (starting 35 days after the beginning of treatment). CLM3 significantly decreased the VEGF-A gene expression in the AF-cell line and the VEGF-A protein and microvessel density in AF tumor tissues.[br]Conclusions. The antitumor and antiangiogenic activity of a new [ldquo]pyrazolo[3,4-d]pyrimidine[rdquo] compound (CLM3) is very promising in anaplastic thyroid cancer, opening the way to a future clinical evaluation.[br][br]Nothing to Disclose: AA, GB, CLM, SMF, PF, ADD, IR, GM, RD, FDS, PM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1445 185 1334 SUN-280 PO44-01 Sunday 1130 2012


1128 ENDO12L_SUN-281 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) A Phase I Study of Cediranib (CED) and Lenalidomide (LEN) in Patients (pts) with Advanced Differentiated Thyroid Cancers (DTCs) [mdash] A Trial of the University of Chicago Phase II Consortium Rebecca L Brown, Theodore G Karrison, Everett E Vokes, Tanguy Y Seiwert, Patricia Heinlen, Jeffrey Bozeman, Walter M Stadler, Ezra E Cohen University of Chicago, Chicago, IL [bold]Background[/bold]: Single agent VEGFR inhibitors and LEN have shown activity in advanced DTC, but combination therapy has not been evaluated. The goal of this study was to determine the dose limiting toxicity (DLT) and maximal tolerated dose (MTD) of combination therapy with the VEGFR inhibitor, CED, and LEN in subjects with DTC.[br][bold]Methods[/bold]: Eligible pts were required to have DTC that was refractory to standard therapy, evidence of measurable disease by RECIST, and at least 20% progression of disease within 12 months of enrollment. A 3 + 3 dose escalation design was employed. Dose escalation was halted when a DLT was observed in the first cycle. DLT was defined as grade (G) 4 hematologic toxicities, G3/4 non-hematologic toxicities except untreated nausea/vomiting or hypersensitivities, or delay in administration of CED or LEN for greater than 2 weeks due to toxicity. Each cycle consisted of 4 weeks of therapy without interruption.[br][bold]Results[/bold]: 18 pts (10 papillary, 4 poorly differentiated papillary, 3 follicular, 1 H[uuml]rthle cell) were enrolled in 3 cohorts: CED 20 mg daily + LEN 15 mg d1-d21 (Cohort 1), CED 30 mg daily + LEN 15 mg d1-21 (Cohort 2), or CED 30 mg daily + LEN 15 mg daily (Cohort 3). 1 pt withdrew prior to initiating therapy and 2 pts did not complete the first cycle (1 pt was withdrawn and 1 pt was diagnosed with new brain metastases) and were therefore not evaluable. DLTs included G3 fatigue in 2 pts (1 in Cohort 2, 1 in Cohort 3) and G3 mucositis in1 pt (Cohort 3). Cohort 2 was expanded to 6 subjects without further DLT so this was determined to be the MTD and recommended Phase II doses. Fatigue (n = 14) and mucositis (n = 7) were the most common G3 toxicities reported. Other G3/4 toxicities included neutropenia (n = 4), anorexia (n = 3), and nausea (n = 2). 14 pts required at least one dose reduction due to toxicity in later cycles. 4 pts discontinued therapy due to toxicity. To date, of 15 evaluable pts, partial response and stable disease, as best response by investigator report, was observed in 3 pts (20%) and 12 pts (80%), respectively. 8 pts have remained on therapy for [gt] 7 months.[br][bold]Conclusions[/bold]: Combination CED and LEN is tolerable with manageable toxicities and demonstrated early evidence of activity in advanced thyroid cancer. A multi-center Phase II study comparing combination CED and LEN to CED alone is underway.[br][br]Sources of Research Support: NCI grant N01-CM-62201.[br][br]Nothing to Disclose: RLB, TGK, EEV, TYS, PH, JB, WMS, EEC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1741 185 1335 SUN-281 PO44-01 Sunday 1131 2012


1129 ENDO12L_SUN-282 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) First Turkish Family with p.A149S Mutation in von Hippel-Lindau Disease Turkan Mete, Dilek Berker, Serhat Isik, Yavuz Yalcin, Ayse Arduc, Oya Topaloglu, Engin Yilmaz, Serdar Guler Ankara Numune Training and Research Hospital, Ankara, Turkey; Hacettepe University Faculty of Medicine, Ankara, Turkey [bold]Objective:[/bold] von Hippel-Lindau (VHL) disease is an autosomal dominant disorder that is characterized by retinal angiomas, renal and pancreatic cysts and carcinomas, pheochromocytomas, epididymal cystadenomas, and variety of multiple tumors. VHL tumor suppressor gene resides on chromosome 3p25-26. The present study aimed to investigate the clinical features and related genetic analysis in a Turkish family with VHL disease.[br][bold]Methods:[/bold] We evaluated members of a family with pheochromocytoma between March 2008 and June 2010. All members were scanned for pheochromocytoma associated other diseases. Genetic analysis for VHL disease was performed by Medical Biology and Genetic Department in Hacettepe University.[br][bold]Results:[/bold] A 35-year- old male patient was admitted to Endocrinology and Metabolism Department. Bilateral pheochromocytoma was diagnosed by necessary tests. We could not detect medullary thyroid carcinoma or hyperparathyroidism. His family history included sudden death in grandfather (age 38) and bilateral renal cell carcinoma (RCC) in father. Then, 16 family members were screened. We detected benign and/or malignant tumors in 11 subjects. These tumors were as follows: Six subjects had bilateral pheochromocytomas, 3 had unilateral pheochromocytoma, 1 had nonfunctional adrenal adenoma and the last one had lumbar spinal hemangioma. Five subjects had only pheochromocytoma, 2 had pheochromocytoma and RCC, 2 had pheochromocytoma with RCC and pancreatic neuroendocrine tumor, one subject had lumbar spinal hemangioma and pancreatic neuroendocrine tumor and one subject had incidental adrenal adenoma with negative VHL gene. Ten of these 11 subjects with tumors had guanine to threonine mutation at nucleotide 658 in second exon. In these subjects the GCC triplet sequence encoding alanine at codon 149, was altered to TCC encoding the serine (p.A149S mutation). Firstly this mutation was reported by Atuk et al in 1998 in a family called Mc C. Their patients had early occurence of retinal angiomatosis but in our family only three patients had retinal anjiomatosis.[bold]Conclusion:[/bold] VHL disease characterized by familial pheochromocytoma together with RCC mostly show this genetic mutation. This is the initial in Turkish population with VHL disease. The present data shows that this mutation indicates more aggressiveness of the disease for our population. We continue to evaluate the other asymptomatic family members for genetic analysis.[br][br]Nothing to Disclose: TM, DB, SI, YY, AA, OT, EY, SG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2244 185 1336 SUN-282 PO44-01 Sunday 1132 2012


1130 ENDO12L_SUN-283 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) An Ectopic ACTH-Producing Lung Carcinoid Detected Only by [sup]68[/sup]Ga-DOTANOC PET-CT Rosa Maria Paragliola, Maria Pia Ricciato, Vittoria Rufini, Guido Rindi, Alfredo Pontecorvi, Salvatore M Corsello Catholic University School of Medicine, Rome, Italy; Catholic University School of Medicine, Rome, Italy; Catholic University School of Medicine, Rome, Italy Cushing[apos]s syndrome (CS) is certainly one of the most challenging endocrine disorders. The greatest problem involves the differentiation between Cushing[apos]s disease and ectopic ACTH syndrome which requires the integration of clinical, biochemical and imaging evaluations. An other challenge is to localize the ACTH-producing source even if the biochemical evaluation points to a sure origin of the ACTH tumor (pituitary or ectopic). A 47 year-old woman was evaluated in another hospital because of ACTH-dependent CS (Urinary cortisol: 1350 mcg/day; ACTH: 97 pg/ml; serum cortisol after 1 mg overnight dexamethasone suppression test: 188 ng/ml). All biochemical and imaging evaluations were suggestive of an ectopic source of ACTH. In particular, serum cortisol after 8 mg dexamethasone suppression test was 170 ng/ml while ACTH and cortisol response to CRH were absent. However, pituitary MRI showed a left microadenoma while negative bilateral inferior petrosal sinus sampling after CRH stimulation ruled out a Cushing disease. The patient underwent two total-body CT and one [sup]111[/sup]In-pentetreotide scan, without any evidence of ACTH secreting ectopic tumor. Bilateral adrenalectomy was then proposed. At that time, she came to our attention. In order to find the ectopic tumor, we performed a total body [sup]68[/sup]Ga-DOTANOC PET-CT showing a focal uptake which corresponded to a 15 mm tumor localized in the left lung lower lobe. Thus, the patient underwent left inferior lobectomy. Histology confirmed a typical bronchial carcinoid (cells were positive for chromogranin A, synaptophysin, and ACTH; Ki67 index was 2.5%). After surgery, serum ACTH and cortisol values fell (ACTH [lt] 5 pg/ml; serum cortisol 30 ng/ml) confirming the remission. The patient is now on oral hydrocortisone and is carefully evaluated in order to gradually reduce the substitutive therapy until the withdrawal. This represents an interesting case of typical bronchial ACTH-secreting carcinoid detected only by [sup]68[/sup]Ga-DOTANOC PET-CT. [sup]18[/sup]-FDG PET CT usually shows low uptake and has low sensitivity for these tumors. Conversely, the few reports on the use of the [sup]68[/sup]Ga-PET are encouraging. We think that [sup]68[/sup]Ga-PET represents an important tool in the diagnostic work-up of patients with ACTH ectopic syndrome and should be more widely used.[br][br]Nothing to Disclose: RMP, MPR, VR, GR, AP, SMC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1126 185 1337 SUN-283 PO44-01 Sunday 1133 2012


1131 ENDO12L_SUN-284 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Asymmetrically Dividing Cells Are the Likely Source of Cancer Stem Cells in Human Anaplastic Thyroid Cancer Risheng Ma, Rauf Latif, Terry F Davies James J Peter VA Medical Center and Mount Sinai School of Medicine, New York, NY Normal and malignant thyroid tissues contain cells with stem cell characteristics as evidenced by the expression of stemness genes such as Oct 4, Nanog and CD-44. The cancer stem cell (CSC) hypothesis postulates that cancers are hierarchically organized and only a subset of cells play a crucial role in tumor initiation, invasive growth, and establishment of metastases. CSCs have been shown to have the ability to self-renew through symmetric and asymmetric cell division and may arise from mutations within an intrathyroidal stem cell population or via epithelial-mesenchymal transition (EMT). Although CSCs have also been detected in thyroid carcinoma cell lines, their origin is unclear. To try and clarify this, we have used an anaplastic thyroid carcinoma cell line (T238) (Rodriguez et al, Brit. J. Cancer 1996), with a doubling time of 19.7 hours, and by pulse-chase experiments with BrdU could detect a cell population that were label-retaining cells ([sim]6%) suggesting the presence of CSCs. These cells divided their DNA both symmetrically and asymmetrically in a ratio of 7:1 and [sim]3% of cells expressed the progenitor marker [ndash] stage-specific embryonic antigen 1 (SSEA-1) which was also expressed asymmetrically (see Figure). The expression of additional stem cell markers (Oct4, sox2, and nanog) was significantly increased in this SSEA-1 positive cell population confirming their stemness potential. However, these cells also showed evidence of EMT initiation with enhanced vimentin expression and decreased E-cadherin expression (259.46% and 0.57% respectively) when compared to SSEA-1 negative cells.[br]These data provide definitive evidence that cellular diversity in anaplastic thyroid cancer may occur through both symmetric and asymmetric cell division. Furthermore, it is likely that the asymmetrically dividing cells have arisen via EMT and may be the source of malignant thyroid tumor formation.[br][br]Nothing to Disclose: RM, RL, TFD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1646 185 1338 SUN-284 PO44-01 Sunday 1134 2012


1132 ENDO12L_SUN-285 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Mutations Confer the mTOR Gene Gain-of-Function and Oncogenecity: Implications for Thyroid Cancer Avaniyapuram Kannan Murugan, Ali Alzahrani, Mingzhao Xing The Johns Hopkins University School of Medicine, Baltimore, MD The phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR)/p70S6K signaling pathway is commonly genetically deregulated in thyroid cancer. The mTOR in this pathway is a serine/threonine protein kinase which regulates cell growth, proliferation, survival and autophagy. mTOR is overactivated in diverse human cancers including thyroid cancer and is being preclinically actively studied as a potential major therapeutic target for thyroid cancer. The mTOR/p70S6K signaling is over-activated mainly by the upstream PI3K/Akt signaling pathway deregulated by various activating genetic alterations in thyroid cancer. However, whether the human mTOR gene itself is a proto-oncogene remains unknown. To answer this fundamental question, we mutated evolutionarily conserved amino acids and generated eight mutants in the HEAT repeats (M938T), FAT (W1456R and G1479N), and kinase (P2273S, V2284M, V2291I, T2294I, and E2288K) domains of mTOR and studied their oncogenicity. Transient expression of these eight mTOR mutants in HEK293T cells showed dramatically elevated protein kinase activities which strongly activated the mTOR/p70S6K signaling. We selected P2273S and E2288K, the two most catalytically active mutants, to further examine their oncogenicity. Stable expression of the two mTOR mutants in NIH3T3 cells strongly activated the mTOR/p70S6K signaling, induced cell transformation and invasion, and caused rapid tumor formation and growth in athymic nude mice after subcutaneous inoculation of the stably-transfected cells. Thus, these mutations conferred mTOR remarkable gain-of-function and oncogenecity. By selectively analyzing 30 exons of mTOR gene for mutations in 12 human thyroid cancer cell lines, we found a natural novel mTOR mutation, resulting in codon and amino acid changes, which we are currently examining for its oncogenicity.[br]This study for the first time demonstrates the oncogenicity of mTOR and establishes the mTOR as a proto-oncogene. By demonstrating the power of strong activation of mTOR to cause tumorigenesis, this study also supports that mTOR is capable of being in a pivotal position to relay the oncogenic signals from the upstream PI3K/Akt pathway in thyroid cancer. These results have important implications for mTOR as a cancer molecule and as a therapeutic target in thyroid cancer.[br][br]Nothing to Disclose: AKM, AA, MX 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2112 185 1339 SUN-285 PO44-01 Sunday 1135 2012


1133 ENDO12L_SUN-286 POSTER SESSION: Neoplasia of Endocrine Tissues (1:30 PM-3:30 PM) Yin Yan Role of Estrogenic Signaling in Human Thyroid Carcinoma Cell Growth: Concomitant Upregulation of Estrogen Receptor-, 12-, 15-Lipoxygenase and 1a- Hdroxylase 25- Hydroxy Vitamin D mRNA Expression Dalia Somjen, Meital Grafi-Cohen, Fortune Kohen, Orli Sharon, Esther Knoll, Zaki Kreim, Naftali Stern Tel Aviv Sourasky Medical Center and Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel; The Weizmann Institute of Science, Rehovot, Israel Estrogen receptors (ERs) are expressed in various cancer cells unrelated to the reproductive system. Some cancer types also express 1a- hydroxylase 25- hydroxyvitamin D (1OHase) mRNA such that the 1,25(OH)[sub]2[/sub]D[sub]3 [/sub](1,25) formed can inhibit tumor cell proliferation via autocrine/paracrine routs. Finally, lipoxygenase (LO) enzyme expression has been detected in pancreatic and other cancer cell types, and their products 12 and 15hydroxyeicosatetraenoic acid (HETE) can enhance tumour cell survival. Since estradiol-17b (E[sub]2[/sub]) enhances thyroid cancer cell growth, we investigated whether or not the estrogenic compounds E[sub]2[/sub], biochainin A (BA), the carboxy- derivative of BA (cBA) as well as the androgen dihydrotestosterone (DHT) affect these systems in the human thyroid carcinoma cell lines. All cell lines expressed ERa, ERb, 1OHase, 12LO and 15LO mRNA (real time PCR). There was a general abundance of ERb over ERa mRNA expression: ERb to ERa mRNA ratio was [gt]1000:1, 228:1 and 7.7:1 in ARO, MRO and NPA cells, respectively. All compounds up-regulated ERb mRNA in ARO, MRO and NPA. All cell lines expressed 1OHase which was up-regulated by all compounds in NPA, by DHT and BA in ARO and by DHT, BA and cBA in MRO. ARO and NPA expressed 15LOmRNA and MRO 12LOmRNA, which were up-regulated by all compounds in NPA, by DHT and BA in ARO and by DHT, BA and cBA in MRO. In parallel, 15 or 12 HETEs were also up-regulated by all compounds. In conclusion thyroid cancer cell lines express mRNA for ERs, 1OHase, 12LO and 15LO which are hormonally modulated. This might form the basis for the use of cancer type-specific estogen/androgen agonists/antagonists and affinity drug targeting, especially via ERb, whose blockade was shown by our group to induce thyroid cancer cell apoptosis (JSBMB 126: 95-103, 2011). These results are consistent with the hypothesis that endogenous estrogens may affect thyroid cancer cell growth via opposing pathways: cell growth acceleration via induction of ER and LO expression, in association with the induction of 1OHase to promote the synthesis of 1, 25 which is known to inhibit cell proliferation.[br][br]Nothing to Disclose: DS, MG-C, FK, OS, EK, ZK, NS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 343 185 1340 SUN-286 PO44-01 Sunday 1136 2012


1134 ENDO12L_SUN-313 POSTER SESSION: Health Education, Outcomes [amp] Comparative Effectiveness (1:30 PM-3:30 PM) How Different Were They? A Retrospective Study of Real-World U.S. Patients with Type 2 Diabetes Mellitus (T2DM) Initiating Once-Daily Injectable Pen Therapy with Insulin Glargine or Liraglutide Sarah Thayer, Wenhui Wei, Lee Brekke, Erin Buysman, Wenli Hu, Robert Cuddihy OptumInsight, San Francisco, CA; sanofi-aventis US, Bridgewater, NJ; OptumInsight, Eden Prairie, MN; sanofi-aventis US, Bridgewater, NJ Initiation of injectable therapy is a major event for T2DM patients but has limited real-world data. Baseline patient characteristics may influence choice of therapy, making comparison of outcomes challenging. This study compared baseline characteristics and described real-world practice patterns and outcomes in adult T2DM patients on oral antidiabetic drugs only, who initiated once-daily injectable therapy using a pen with insulin glargine (GLA-P) or liraglutide (LIRA), a glucagon-like peptide-1 analog.[br]This retrospective study used a claims database from a large US health plan affiliated with OptumInsight. Patients initiated GLA-P or LIRA between January and July 2010 and had continuous health coverage for 6 months before (baseline) and 9 months after (follow-up). Differences in baseline characteristics between the cohorts were tested. 9-Month follow-up measures (A1C, treatment persistence and health care costs) were assessed descriptively.[br]A total of 975 unmatched T2DM patients (GLA-P n=610; LIRA n=365) were included. Significant differences existed between the cohorts at baseline. Compared with LIRA patients, GLA-P patients were more often male (57.5% vs 50.7%, [italic]P[/italic]=0.039), older (56.2 vs 52.8 years, [italic]P[/italic][lt]0.001), less obese (11.8% vs 17.8%, [italic]P[/italic]=0.01), more often on sulfonylureas (68.4% vs 43.6%, [italic]P[/italic][lt]0.001), and had more comorbidities (Charlson comorbidity index 1.22 vs 0.62, [italic]P[/italic][lt]0.001). A higher percentage had [ge]1 hospitalization (11.5% vs 2.5%, [italic]P[/italic][lt]0.001), higher baseline A1C (9.7% vs 7.9%, [italic]P[/italic][lt]0.001), and A1C [ge]9.0% (58.7% vs 21.6%, [italic]P[/italic][lt]0.001). More LIRA patients had A1C [lt]7.0% (8.9% vs 29.6%, [italic]P[/italic][lt]0.001).[br]During follow-up differences in the unmatched sample remained. GLA-P patients continued to have a higher mean A1C (8.4% vs 7.4%), but greater A1C reduction ([minus]1.2% vs [minus]0.6%). Treatment persistence was 63.1% (GLA-P) and 52.1% (LIRA); study drug costs contributed 6.6% (GLA-P) and 17.0% (LIRA) of mean total health care costs of $12,420 and $11,238 respectively.[br]There were significant differences at the time of initiating GLA-P or LIRA as the first injectable therapy. GLA-P initiators had more comorbidities and higher utilization. Almost 1/3 of LIRA initiators had baseline A1C [lt]7.0%, suggesting different treatment priorities for the 2 cohorts following first FDA approval of LIRA in 2010. These findings suggest that LIRA and GLA-P prescribing patterns are influenced by baseline characteristics, presenting challenges in conducting related comparative effectiveness research.[br][br]Sources of Research Support: Study funding and editorial support provided by sanofi-aventis U.S.[br][br]Disclosures: ST: Employee of OptumInsight which is under contract with sanofi-aventis U.S., Sanofi-Aventis. WW: Employee, Sanofi-Aventis; Stockholder, Sanofi-Aventis. LB: Employee of OptumInsight which is under contract to sanofi-aventis U.S, Sanofi-Aventis. EB: Employee of OptumInsight which is under contract to sanofi-aventis U.S., Sanofi-Aventis. WH: Employee, Sanofi-Aventis. RC: Employee, Sanofi-Aventis. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1762 186 1341 SUN-313 PO15-01 Sunday 1137 2012


1135 ENDO12L_SUN-314 POSTER SESSION: Health Education, Outcomes [amp] Comparative Effectiveness (1:30 PM-3:30 PM) Glycemia in Insulin-Requiring Older Adults Residing in a Skilled Nursing Facility Pearl K Dy, Philip C Morin, Ruth S Weinstock State University of New York (SUNY) Upstate Medical University, Syracuse, NY The prevalence of diabetes in older adults is increasing, with many requiring insulin therapy. Insulin therapy in residents of skilled nursing facilities (SNF) poses unique challenges and has not been well studied.[br]Many residents of SNFs have multiple comorbidities, variable and unpredictable food intake, poor mobility, poor balance with increased risk for falls, incontinence, depression and inability to communicate hypoglycemia. Goals include avoidance of both hypoglycemia and serious or symptomatic hyperglycemia.[br]Data were collected retrospectively from a SNF for a 30 day period (November, 2011) for residents receiving insulin therapy (n=14; 4 males, 10 females) after obtaining informed consent from the patient and/or legal guardian: resident mean (range) age 83 yrs (73-93 yrs), BMI 28 kg/m[sup]2[/sup] (20-38 kg/m[sup]2[/sup]), A1c 8.2% (6.6-12.6%). Half were not alert and oriented (6 confused, 1 aphasic). Mean daily meal intake was 73% (38-89%), but for individual meals intake ranged from 0% to 100%. Lowest breakfast, lunch and dinner consumption were 33%, 28% and 56% respectively.[br]Insulin regimens were: oral medication with pre-mealtime insulin (n=4, 29%), basal with pre-meal insulins (n=3, 21%), oral medication with basal and premeal insulin (n=3, 21%), oral agent with basal insulin (n=1, 7%), premixed insulin (n=1, 7%) and premixed insulin with mealtime insulin (n=2, 14%).[br]Capillary glucose measurements were mostly taken 2-4 times daily. There were 13 measurements [le] 80 mg/dl in 7 residents (50% of residents), 157 glucose levels [ge] 250 mg/dl in 10 residents (71% of residents), and 4 residents (29%) had levels [ge] 400 mg/dl. The resident with the greatest number of glucose values [lt]80 mg/dl had an A1c of 9.1%; the 2 residents with the greatest number of glucose levels [gt]350 mg/dl had A1cs of 7.7% and 9.4%. No residents had their insulin therapy orders modified during the month, and none had their mealtime insulin dose adjusted based upon their food (carbohydrate) intake. Only 2 residents (14%) had all their values between 80 and 250 mg/dl over the month.[br]Although this pilot study is small, the data suggest that there are opportunities to improve insulin management and glycemic control for residents in SNFs.[br][br]Nothing to Disclose: PKD, PCM, RSW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1015 186 1342 SUN-314 PO15-01 Sunday 1138 2012


1136 ENDO12L_SUN-315 POSTER SESSION: Health Education, Outcomes [amp] Comparative Effectiveness (1:30 PM-3:30 PM) Characteristics of High Utilization Claims for Home Blood Glucose Test Strips and Lancets for Medicare Beneficiaries Yun Suk Kim, Monica Adriana Mejia, Janet Amoy McLeod, Elizabeth Ann Koller US Department of Health and Human Services, Monterey Park, CA; US Department of Health and Human Services, Baltimore, MD; Centers for Medicare and Medicaid Services, Baltimore, MD [bold]BACKGROUND: [/bold]Medicare coverage criteria permit payment for 100 blood glucose test strips and lancets per month for insulin (I)-using beneficiaries and the same amount per 3 months for non-I-using beneficiaries. When beneficiaries need supplies in excess of criteria, physicians and vendors of durable medical equipment are required to maintain substantiating documentation. Supplies are to be refilled only near a prescription[apos]s end and if requested by the beneficiary. This study was conducted to assess whether the criteria for high utilization claims for test strips and lancets were met and to characterize any deficiencies.[br][bold]METHODS: [/bold]Medicare high utilization claims were randomly selected. Testing frequency by treatment type, overlapping service dates, and documentation adequacy were assessed.[br][bold]RESULTS: [/bold]Of 400 claims, 319 and 81 were for test strips and/or lancets in non-I and I-treated beneficiaries, respectively. Only 97 were in compliance. 303 claims (76 %) had [ge]1 deficiencies; 182/303 claims contained [gt]1 deficiency. Deficiencies included lack of: a) documentation of the rationale for excess utilization (n=222), b) documentation in support of refills (n=117), c) complete physician orders (n=90), and d) proof of delivery by the supplier (n=33). For 20 claims without rationale for frequent testing, there was also no evidence that the prescriber had seen the patient in the prior 6 months. Of claims lacking documentation for refills, in 36 no refill had been requested and, in 91 refills were dispensed prior to near depletion. Supplies were provided by multiple vendors to the same beneficiaries for 56 claims[italic]. [/italic][br]58% of claims for non-I-treated beneficiaries lacked documentation to support excess utilization. In 77, there was no justification for [gt]2x/d testing (and as frequent as 8x/d). In 79, there was no justification for the prescribed 2x/d testing. For 30, a physician order or specific testing frequency in the order was missing. 44% of claims for I-treated beneficiaries lacked documentation to support excess utilization. In 27, there was no justification for testing [gt]3x/d. In 1, there was documentation to support 2x/d, but not 3x/d testing. For 8, a physician order or specific testing frequency in the order was missing.[br][bold]CONCLUSIONS: [/bold]Poor documentation for supplies in excess of coverage criteria, duplicate supplies, and unrequested refills are major reasons for claims to be classified as noncompliant. Findings were similar for all 4 regional Medicare contractors.[br][br]Nothing to Disclose: YSK, MAM, JAM, EAK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1854 186 1343 SUN-315 PO15-01 Sunday 1139 2012


1137 ENDO12L_SUN-316 POSTER SESSION: Health Education, Outcomes [amp] Comparative Effectiveness (1:30 PM-3:30 PM) Association of Preoperative and Postoperative Glucose Control with Complications among Diabetic Patients M Kathleen Figaro, Jeffrey B Boord, Michael E Matheny Vanderbilt University Medical Center, Nashville, TN; VA Tennessee Valley Health System, Nashville, TN; Vanderbilt University Medical Center, Nashville, TN OBJECTIVE: Examine the relationship between preoperative and postoperative glucose control and postoperative complications in a multicenter sample of hospitalized diabetic patients undergoing elective surgical procedures. Perioperative glucose control has been linked to postoperative complications. Less is known about preoperative control and its impact on postoperative course.[br]METHODS: Retrospective analysis of 1,430 hospitalized diabetic patients undergoing elective surgeries in 4 Veterans Heath Administration hospitals from 2002 to 2009. Data were collected from laboratory data and inpatient/outpatient administrative codes. Multivariate regression was performed of a composite postoperative complication comprising wound infection, acute kidney injury, systemic infection, vascular complications and death adjusting for demographics, preoperative A1C, diabetes duration, and mean serum glucose within 72 hours after surgery.[br]RESULTS: The average age was 64.7(9.8) years. Patients underwent 18 types of elective procedures including colectomy, other gastrointestinal surgeries, vascular procedures, lobectomy, fracture repair, hernia repair, prostatectomy, spinal fusion and total Joint replacement. The most common procedures were total joint replacement (20.3%), vascular procedures (9.2%), colectomy (9.2%) and prostatectomy (8.7%). Mean (SD) preoperative A1C was 7.2% (1.5).More than 60% of patients had preoperative A1C [gt]8%. 29% and 7% of patients had a mean 72 hour postoperative glucose concentration at least 150 or 200 mg/dL, respectively. The overall postoperative complication rate was 21%. Mean 72 hour postoperative serum glucose concentrations were not associated with higher rates of postoperative complications in either the bivariate or multivariate analyses; (OR: 1.001; 95% confidence interval,.99-1.004; P =.47).[br]CONCLUSIONS: In a preliminary analysis of a multi-center sample of diabetic patients undergoing a variety of elective surgical procedures, average A1c within the last 6 months and glucose control over 72 hours after surgery were not predictive of complications or mortality. However, different types of elective procedures may be associated with different predictors of postoperative complications.[br][br]Sources of Research Support: This material is the result of work supported with resources and the use of facilities at the VA Tennessee Valley Healthcare System.We would additionally like to acknowledge the Vanderbilt University Medical Center endocrinology fellowship and Michael Matheny[apos]s VA HSR[amp]D CDA-2 08-020 for supporting the work leading to this abstract.[br][br]Nothing to Disclose: MKF, JBB, MEM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1800 186 1344 SUN-316 PO15-01 Sunday 1140 2012


1138 ENDO12L_SUN-317 POSTER SESSION: Health Education, Outcomes [amp] Comparative Effectiveness (1:30 PM-3:30 PM) Improving Comprehensive Diabetes Care in Skilled Nursing Facilities: Lessons from a Quality Improvement Initiative Patrick J Boyle, Sara C Miller, Kevin O[apos]Neil University of New Mexico Health Sciences Center, Albuquerque, NM; Med-IQ, LLC, Baltimore, MD; Brookdale Senior Living, Brentwood, TN [bold]BACKGROUND: [/bold]The growing population of older adults with diabetes residing in long-term care (LTC) communities is an especially vulnerable subset of patients. Disease management is further complicated by substantial numbers of transitions between care settings, care provision from multiple providers, and cognitive limitations. We report the results of a quality improvement initiative focused on improving the care for residents with diabetes in LTC communities.[br][bold]METHODS: [/bold]Quantitative data were collected through chart abstraction using predetermined measures, qualitative data used focus-group interviews. Targeted education through a live CE/CME-certified seminar was provided to nursing assistants, nurses, and physicians. Learnings from the baseline data and live education were extended across the Brookdale network through a CE/CME-certified monograph and included tools for the implementation of improvement efforts. Effects of the education were assessed through chart reviews at 3 and 5 months post-education.[br][bold]RESULTS: [/bold]Performance of comprehensive foot evaluations was reported for 43% of patients at baseline and increased to 60% and 56% at 3 and 5 months post-education. Referrals to specialists when foot evaluations were abnormal increased from 50% at baseline to 92% and 80% in follow-up abstractions. Provision of eye exams increased (46% at baseline, 66% and 55% at follow-up). Notable reductions in LDL-C were seen; mean at baseline was 93 and reduced to 87 and 83 at follow-up. Mean A1C values were unchanged but, there was a notable reduction in the range (5.1% to 12.2% at baseline; 5.4% to 10.4%, and 5.4% to 9.9% at 3 and 5 month follow-up) and reductions in daily blood glucose values measured before lunch (baseline mean 208 mg/dL reduced to 194 mg/dL and 192 mg/dL), before dinner (mean 199 mg/dL, reduced to 170 mg/dL and 181 mg/dL) and before bed (229 mg/dL reduced to 174 mg/dL and 195 mg/dL). Percentage of residents with no hypoglycemic events (self-report or recorded blood glucose) increased from 69% at baseline to 88% and 89% at each follow-up. No improvements were seen in BP, documentation of some aspects of care (lipid panel, A1C values), use of sliding-scale insulin, or documentation of orders for time between insulin and meals.[br][bold]CONCLUSIONS: [/bold]Education efforts which include a focus on identified areas of need represent an effective means for improving care of elderly patients with diabetes, despite the significant complexity of caring for these residents.[br][br]Sources of Research Support: Financial support for data collection, analysis and CME-activity provided through an unrestricted educational grant from sanofi awarded to Med-IQ. Funding organization played no role in data collection, analysis and presentation. Funder also had no role in the development and presentation of the CME-activity.[br][br]Disclosures: PJB: Advisory Group Member, Merck & Co.; Speaker, Eli Lilly & Company, Amylin Pharmaceuticals, Takeda. Nothing to Disclose: SCM, KO 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2165 186 1345 SUN-317 PO15-01 Sunday 1141 2012


1139 ENDO12L_SUN-318 POSTER SESSION: Health Education, Outcomes [amp] Comparative Effectiveness (1:30 PM-3:30 PM) Pre-Index Clinical Differences between Type 2 Diabetes Patients with Renal Impairment Prescribed Saxagliptin and Other Non-Insulin Antidiabetic Medications Sarah Thayer, Setareh Williams, Ying Fan, Shreekant Parasuraman OptumInsight, San Francisco, CA; AstraZeneca, Wilmington, DE; OptumInsight, Eden Prairie, MN [bold]Background:[/bold] In claims database studies, reasons for treatment initiation are generally unobservable. Patients[apos] clinical characteristics may influence choice of therapy, especially in populations at risk for poor outcomes. This study compared pre-index characteristics of type 2 diabetes mellitus (T2DM) patients with renal impairment (RI) initiating the dipeptidyl peptidase-4 inhibitor saxagliptin (SAXA) versus those initiating other non-insulin antidiabetic (AD) therapy.[br][bold]Methods:[/bold] Individuals age [ge]18 years and with evidence of T2DM (ICD-9-CM 250.x0 or 250.x2) were identified from a large nationally representative US health plan. Patients with [ge]1 pharmacy claim for SAXA between 8/1/2009 and 12/31/2010 were assigned to the SAXA cohort, and patients with [ge]1 pharmacy claim (8/1/2009-12/31/2010) for other oral AD medications or GLP-1 analogs were assigned to the Other cohort. The first claim for SAXA or another study drug was the index date. Patients were required to be naive to SAXA or the Other regimen for 12 months before the index date. RI was determined from lab data before the index date, and was defined (mild, moderate, or severe) as a glomerular filtration rate [lt] 90 mL/min/1.73 m[sup]2[/sup]. Variables were measured during a 12-month (pre-index) period before treatment initiation.[br][bold]Results:[/bold] Of all T2DM patients identified, a significantly higher percentage initiating SAXA vs. Other had RI (30.3% vs 26.5% p[lt]0.001). In total, 615 T2DM patients with RI were included in the SAXA cohort, and 6294 in the Other cohort. RI tended to be more severe among SAXA patients versus Other patients. Patients who initiated prescribed SAXA treatment had significantly higher pre-index mean Charlson-Quan comorbidity scores versus Other patients (2.5 vs 2.3; [italic]P[/italic]=.004), and SAXA patients had significantly higher pre-index rates of retinopathy/eye complications, erectile dysfunction/impotence, and neuropathy (all [italic]P[/italic][lt].001). Also, patients initiating SAXA had higher pre-index HbA1c levels (7.9% vs 7.5%; [italic]P[/italic][lt].001) and fewer had pre-index glycemic control (HbA1c[lt]7%) compared with Other patients (28.2% vs 46.8%; [italic]P[/italic][lt].001).[br][bold]Conclusions:[/bold] Patients who were prescribed SAXA treatment appeared to have worse health status before initiation of therapy than those initiating other AD therapy. These findings suggest the SAXA prescribing pattern is influenced by multiple factors that lead to a patient cohort with distinctly different clinical characteristics than patients prescribed other AD agents.[br][br]Sources of Research Support: Astra Zeneca.[br][br]Disclosures: ST: Employee, OptumInsight. SW: Employee, Astra Zeneca. YF: Employee, OptumInsight. SP: Employee, Astra Zeneca. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 649 186 1346 SUN-318 PO15-01 Sunday 1142 2012


1140 ENDO12L_SUN-319 POSTER SESSION: Health Education, Outcomes [amp] Comparative Effectiveness (1:30 PM-3:30 PM) Increasing Awareness of the Hormonal and Fertility Impacts of Cancer Kate E Waimey Timmerman, Lorrie Fritz, Teresa K Woodruff Feinberg School of Medicine, Northwestern University, Chicago, IL; The Hormone Foundation, Chevy Chase, MD Every year, more that 135,000 young people are diagnosed with cancer and many survive their disease. Survivors experience long term or late effects after treatment, including those due to reduced function of endocrine glands such as the testes, ovaries or pituitary, and may result in low testosterone, decreased libido, and erectile dysfunction in men, and early menopause, leading to hot flashes, vaginal atrophy, sleep disturbances and osteoporosis in women. Childhood growth and pubertal development can be compromised. Cancer or its treatment may also cause infertility and recommendations from the American Society of Reproductive Medicine, European Society of Human Reproduction and Embryology (ESHRE) and others state that people should be informed about the endocrine and reproductive risks of cancer treatment and options but patients still do not receive adequate hormonal and fertility information. Educating patients on symptom management is challenging due to the wide spectrum and difficulty of predicting the exact impact of cancer treatment on reproductive health and function. In June 2011, the Oncofertility Consortium and Hormone Foundation launched a program to educate providers and the public about the fertility and hormonal risks of cancer treatment and options for men, women and children. Print provider guides, a website, and the first mobile application on cancer and fertility were developed. In the first 7 months of the campaign, guides were mailed to more than 19,000 providers, the SaveMyFertility.org website received 3,850 visits and 7,971 page views, and the iSaveFertility iPhone App was downloaded 642 times, with 1,515 uses. Users spend more than 11 minutes on the App and return multiple times, providing some of the first evidence that clinicians may learn new health information through mobile devices. Survey responses indicate that more than 75% and 50% of providers found the guides and fact sheets useful, respectively, while talking with patients. Only 20% of providers reported to print materials off the web to facilitate discussions with patients, indicating that individual preferences and technology usage come into play when educating providers and encouraging them to disseminate materials about endocrine and reproductive health to patients.[br][br]Sources of Research Support: The campaign was supported by grants from Merck [amp] Co., Inc. and EMD Serono. The Oncofertility Consortium is funded by the NIH Roadmap for Medical Research, grant 5 UL1 DE19587-05.[br][br]Nothing to Disclose: KEWT, LF, TKW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 760 186 1347 SUN-319 PO15-01 Sunday 1143 2012


1141 ENDO12L_SUN-320 POSTER SESSION: Health Education, Outcomes [amp] Comparative Effectiveness (1:30 PM-3:30 PM) Correlates of Goiter and Other Iodine-Deficiency Disorders: A Case-Control Study Narendra Kotwal, Jyoti Kotwal, Atul Kkotwal Command Hospital (SC), Pune, India; AFMC Pune, Pune, India Goiter and other Iodine Deficiency Disorders (IDDs) are known to mankind since antiquity. The role of iodine in the causation of goiter is well known However, despite the strategies adopted for the control of IDDs, the problem has increased in magnitude. We focussed our research on the issues of lack of adequate data on thyroid function, focus on monocausality or ignoring multi causality on finding and possible link of increased iodine intake leading to increased thyroid dysfunctions e.g. hypothyroidism, autoimmune thyroiditis. A case control study was conducted with three groups of study participants. The study was conducted at Delhi [amp] Pune. Group (gp) I consisted of cases (patients with established goiter or overt hypothyroidism in various age groups); Group II comprised of age and sex matched hospital controlsand Group III consisted of community based controls. The study participants were administered a nutrition questionnaire including salt intake and socio-demographic factors. Additionally, Thyroid function tests, Microsomal Thyroid antibody and Urinary iodine estimation was carried out for all study participants.[br]A large proportion of cases (67.33%) had hypothyroidism due to autoimmune thyroiditis and all these patients had urinary iodine either adequate (151-299 mg/L) or excessive ([gt] 300 mg/L). Only 5 patients has mild diffuse goiter on clinical examination. Thus the present data on cases have shown that there has been a shift in profile of cases of hypothyroidism from endemic goiter to sporadic goiter i.e. from goiter due to Iodine deficiency to autoimmune thyroiditis and may be due to excess of iodine. A large proportion of hospital controls (49.48%) had urinary iodine in the range which can be harmful ([gt]300mg/L) and also anti-TPO antibodies thereby making them subclinical hypothyroidism due to autoimmune disease probably due to excess Iodine. 69 (14.14%) of community controls had raised anti-TPO and 48.97% of them had urinary iodine in dangerous levels i.e. in excess of 300 mg/L, showing that all had adequate or more than adequate Iodine in their diet. There was a statistically significant difference between cases and controls in the median values of TSH, anti-TPO and urinary iodine (p[lt]0.05).[br]The study has clearly shown that cases of hypothyroidism are occurring due to excess Iodine intake. An eco-social epidemiological approach has been suggested as a way forward.[br][br]McCarrison R. Observation on endemic goiter in the Chitral and Gilgit valleys. Lancet 1906; 1: 1110. 2. McCarrison R. An Experiment in Goitre Prevention being The Further History of Goitre at the Lawrence Royal Military School, Sanawar, Punjab, India. BMJ 1927; January 15: 94-95. 3. Stott H., Bhatia BB Lal RS, Rai KC. The Distribution and Cause of Endemic Goitre in the United Provinces. IJMR 1930; 1059-1085. 4. Ramalingaswami V, Subramanian TAV, Deo M.G. The Aetiology of Himalayan Endemic Goiter. Lancet 1961; March 15: 791-794. 5. Kochupillai N, Karmarkar MG, Weightman, HR, et al. Pituitary-Thyroid Axis in Himalayan Endemic Goiter. Lancet 1973; May 12: 1021-1024.[br][br]Sources of Research Support: DGAFMS office.[br][br]Nothing to Disclose: NK, JK, AK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1226 186 1348 SUN-320 PO15-01 Sunday 1144 2012


1142 ENDO12L_SUN-321 POSTER SESSION: Health Education, Outcomes [amp] Comparative Effectiveness (1:30 PM-3:30 PM) Development of a Thyroid Biopsy Clinic at an Academic Institution Liselle Douyon University of Michigan, Ann Arbor, MI [bold]Introduction:[/bold][br]Ultrasound guided biopsy of the thyroid is a procedure that clinical endocrinologists perform. In an effort to provide training to endocrinology housestaff and to expand biopsy services, a thyroid biopsy clinic was started. A summary of the process is provided.[br][bold] Methods and Materials: [/bold][br]A needs assessment for the clinic was performed. A reliable ultrasound machine and the need to have pathology present at the time of ultrasound guided fine needle aspiration thyroid biopsy(US-FNA)were identified as crucial. Teaching and support staff were from endocrinology and pathology. Learners included endocrine fellows, medical students and medical assistants. Each learner had specific educational goals. A variety of instruction methods were used: didactic lectures, clinical bedside demos, assigned reading, discussions, web based computer education, interactive demos using biopsy models and biopsy guns, microscope, ultrasound and slide making training sessions.[br][bold] Results: [/bold][br]All learners improved their skills and successfully reached their educational targets. The thyroid biopsy clinic was moved off site. With the use of telepathology the cytopathologist was able to review all specimens at the time of the procedure. The number of nondiagnostic biopsies decreased. Clinical biopsy services expanded. Telepathology was an effective teaching tool. Based on the beneficial clinical experience in our clinic, telepathology is being expanded to other medical units.[br][bold] Conclusions: [/bold][br]The establishment of a thyroid biopsy clinic has been a success. It is an effective way to train housestaff and support staff. Biopsy outcomes improved. Clinical services increased. A strong collaboration was established between the endocrinologist and the pathologist.[br][br]Nothing to Disclose: LD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1473 186 1349 SUN-321 PO15-01 Sunday 1145 2012


1143 ENDO12L_SUN-322 POSTER SESSION: Health Education, Outcomes [amp] Comparative Effectiveness (1:30 PM-3:30 PM) Prevalence of Complications and Treatment in Acromegaly Patients in the United States Michael S Broder, Maureen P Neary, Eunice Chang, Dasha Cherepanov, Laurence Katznelson Partnership for Health Analytic Research, LLC, Beverly Hills, CA; Novartis Pharmaceuticals Corporation, East Hanover, NJ; Stanford University, Stanford, CA Objectives: Acromegaly, a rare, slowly progressing disorder resulting from excessive growth hormone, is associated with a variety of complications, including cardiovascular disease, reproductive disorders, and arthropathy. Delayed treatment can lead to worsening of these complications. The frequency of complications and treatment in the general acromegaly population is largely unknown. We describe the prevalence of complications and a variety of treatments in acromegaly patients.[br]Methods: Combining two commercial US claims databases, Thomson Reuters MarketScan and IMS Health PharMetrics, we identified acromegaly patients between 1/1/2003-12/31/2009. Each continuously-enrolled patient was followed for 1 year (review period) after his/her first observed acromegaly claim. All pharmacy and medical claims in the review period were used to estimate prevalence of complications and treatment.[br]Results: We identified 2336 acromegaly patients (mean age: 45.3 years; 50.9% female; mean Charlson comorbidity index: 1.0). Proportions with cardiovascular risk factors were hypertension: 29.6%; hypertriglyceridemia: 19.8%; and diabetes: 17.1%. Proportions with complications were musculoskeletal abnormalities: 25.1%; hypopituitarism: 15.1%; sleep apnea: 11.1%; reproductive system abnormalities: 10.7%; cardiovascular abnormalities: 9.7%; and colon polyps or cancer: 6.3%. During the study period, 5.2% (121) had surgery and 2.4% (57) received radiation therapy. Pharmacologic treatment was used by 39.1% (914), including 18.6% (435) using long-acting release octreotide, 18.2 % (425) using dopamine agonists, 4.5% (104) using short-acting octreotide, 4.1% (96) using pegvisomant, and 1.2% (28) using lanreotide (not mutually exclusive). Fifty-three percent (1,234) of patients had at least one biochemical monitoring test (either IGF-1 or growth hormone) during the year: 50.9% (1,189) of patients had IGF-1 and 30% (700) had growth hormone test in that time period.[br]Conclusions: In this study, cardiovascular risk factors were common in acromegaly patients. Serious comorbidities, including musculoskeletal and reproductive system abnormalities, hypopituitarism, and sleep apnea were also common. Octreotide was the most frequently used drug treatment. Approximately half of patients had no biochemical monitoring in a year. Appropriate attention to these complications along with adequate therapy and monitoring are critical in the approach to this disease.[br][br]Disclosures: MSB: Principal Investigator, Novartis Pharmaceuticals. MPN: Employee, Novartis Pharmaceuticals. EC: Researcher, Novartis Pharmaceuticals. DC: Researcher, Novartis Pharmaceuticals. LK: Coinvestigator, Novartis Pharmaceuticals, Ipsen. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1710 186 1350 SUN-322 PO15-01 Sunday 1146 2012


1144 ENDO12L_SUN-323 POSTER SESSION: Health Education, Outcomes [amp] Comparative Effectiveness (1:30 PM-3:30 PM) Healthcare Utilization and Costs in Acromegaly Patients in the United States Michael S Broder, Maureen P Neary, Eunice Chang, Dasha Cherepanov, Laurence Katznelson Partnership for Health Analytic Research, LLC, Beverly Hills, CA; Novartis Pharmaceuticals Corporation, East Hanover, NJ; Stanford University, Stanford, CA Objectives: Acromegaly, a chronic and debilitating disorder caused by excessive growth hormone secretion, results in considerable comorbidities, declines in quality of life and increased mortality. Many acromegaly patients are not effectively treated and suffer from slowly progressive disease complications. The economic burden of these comorbidities has not been well characterized. We describe healthcare utilization and costs associated with acromegaly in the US.[br]Methods: Combining 2 commercial US claims databases, Thomson Reuters MarketScan and IMS Health PharMetrics, we identified acromegaly patients between 1/1/2003-12/31/2009. Each continuously-enrolled patient was followed for 1 year (review period) after his/her first observed acromegaly claim. All pharmacy and medical claims in the review period were used to estimate utilization and costs. Univariate analyses and regression models were used to model cost and utilization.[br]Results: We identified 2,336 acromegaly patients (mean age: 45.3 yrs.; 50.9% female; mean Charlson comorbidity index: 1.0). Inpatient hospitalization was observed in 16.6% of patients and emergency department visits in 22.4%. Patients had a mean of 15.6 office visits/year. Total healthcare costs were $24,284 (SD: $33,341)/per patient-year (PPY). Of this total, $16,995 (SD: $30,047) was from medical costs and $7,289 (SD: $14,157) from pharmacy costs. After adjusting for age, gender, region and risk factors, the presence of comorbidities significantly increased costs (p[lt].01). Colon polyps or cancer increased cost by $9,697; musculoskeletal abnormalities by $6,916; cardiovascular abnormalities by $13,573; sleep apnea by $10,160; and hypopituitarism by $7,166. Odds of hospitalization increased with musculoskeletal abnormalities (OR:1.6; 95% CI:1.2-2.1), cardiovascular abnormalities (OR:2.9; 95% CI:2.1-3.9), and sleep apnea (OR:1.5; 95% CI:1.1-2.1). Odds of emergency department visit increased with musculoskeletal (OR:1.8; 95% CI:1.4-2.3) and cardiovascular abnormalities (OR:2.4; 95% CI:1.8-3.3).[br]Conclusions: Although acromegaly is rare, complications suffered by acromegaly patients increase both utilization and cost. Cardiovascular complications increased cost among these patients by more than $13,000/year and almost tripled the odds of hospitalization. Since many complications occur slowly over time, earlier and more intensive treatment might reduce them. Future studies should estimate the long-term effect of treatment on costs.[br][br]Disclosures: MSB: Principal Investigator, Novartis Pharmaceuticals. MPN: Employee, Novartis Pharmaceuticals. EC: Researcher, Novartis Pharmaceuticals. DC: Researcher, Novartis Pharmaceuticals. LK: Coinvestigator, Novartis Pharmaceuticals, Ipsen. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1713 186 1351 SUN-323 PO15-01 Sunday 1147 2012


1145 ENDO12L_SUN-324 POSTER SESSION: Health Education, Outcomes [amp] Comparative Effectiveness (1:30 PM-3:30 PM) Improving Quality in Type 2 Diabetes: A National Initiative To Assess Guideline Adherence and Physician/Pharmacist Communication Timothy Reid, James Rosenzweig, Karen Reed, Peter Sheldon Mercy Diabetes Center, Janesville, WI; Boston Medical Center, Boston, MA; Kmart Pharmacy, Beckley, WV; Opus Science, LLC, Columbia, MD [bold]Background: [/bold]Coordination of care is a key component of successful management of patients with type 2 diabetes. In response to this need, a multiphase initiative will compare practice barriers related to caring for patients with type 2 diabetes with an assessment of actual in-practice behavior in multiple sites across the US. This multidisciplinary initiative will be an effective tool for measuring current diabetes management practices among physicians as well as gauging unique systemic and professional barriers to cooperation with local pharmacists.[br][bold]Objective: [/bold]The objectives of this initiative are: 1) to uncover practice-specific barriers to appropriate glycemic control in primary care and endocrinology practices; 2) analyze current pharmacy practices as they relate to medication education and monitoring of patients with type 2 diabetes; 3) determine the critical attributes of successful interdisciplinary diabetes care, involving physician, office staff, and pharmacists; 4) evaluate the impact of providing physicians with feedback regarding diabetes management performance and communication with pharmacists.[br][bold]Methods: [/bold]This mixed-methods study will involve a national survey of PCPs and endocrinologists; seven live, in-practice assessments with physicians, office staff members, and local pharmacists; practice assessments outlining areas for improvement based on expert faculty recommendations and guidelines; and pre-post chart reviews to measure effectiveness. A follow-up survey will measure participant satisfaction with the assessment process and the degree to which practice behaviors have changed as a result of these assessments.[br][bold]Results: [/bold]This study will allow the accurate collection of quantitative and qualitative information on how physician practices operate, and interact with their local pharmacists, in their care of patients with type 2 diabetes and to determine whether using chart review, as well as practice interviews to gather data and subsequently compare to guidelines, is sufficient to prompt a quantifiable behavior change by physicians with respect to diabetes patient management. Preliminary survey and assessment data will be presented.[br][bold]Discussion:[/bold] The program committee focused research on 4 underutilized resources in primary care: patient data; referrals to specialists; local pharmacists; and patients themselves. The resulting data and subsequent faculty recommendations are intended to give PCPs guidance for improving patient outcomes.[br][br]Nothing to Disclose: TR, JR, KR, PS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2059 186 1352 SUN-324 PO15-01 Sunday 1148 2012


1146 ENDO12L_SUN-325 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Safety and Tolerability of Recombinant Human Parathyroid Hormone (rhPTH[1[ndash]84]) in a Randomized, Double-Blind, Placebo-Controlled Study for the Treatment of Adults with Hypoparathyroidism Dolores Shoback, Bart Clarke, Maria Luisa Brandi, John Bilezikian, Hjalmar Lagast San Francisco VA Medical Center, University of California, San Francisco, CA; Mayo Clinic Rochester, Rochester, MN; University Hospital of Careggi, Florence, Italy; Columbia University, New York, NY; NPS Pharmaceuticals, Bedminster, NJ Hypoparathyroidism (HypoPARA) is a rare endocrine disorder where the parathyroid glands never develop normally, get destroyed, or produce insufficient hormone. Therapy is currently limited to controlling symptoms, often with large amounts of calcium (Ca) and active vitamin (Vit) D metabolites resulting in possible long-term complications such as end-organ damage due to renal, ocular, and soft tissue complications.[br]This 24-week, randomized, double-blind, placebo (PBO)-controlled study assessed the safety and efficacy of a flexible-dose (50, 75, 100 [micro]g) daily injection of recombinant human parathyroid hormone (rhPTH[1[ndash]84]) plus optimized oral therapy with Ca citrate and Vit D (calcitriol or alphacalcidol) tablets vs optimized oral therapy alone. Adverse events (AE) were monitored throughout treatment and during a 4-week follow-up period.[br]Of the 196 subjects screened, 134 completed the optimization period and were randomized (2:1) to rhPTH(1[ndash]84) (n=90) or placebo (n=44). Six (7%) subjects in the rhPTH(1[ndash]84) group discontinued treatment, 3 by decision and 3 due to AEs (1 treatment-related): 1 each for stroke, multiple symptoms, and hypertension. Seven (16%) subjects in the PBO group discontinued, 6 by decision and 1 for noncompliance. The median rate of compliance with injections was [gt]99% in both groups.[br]By study end, 84 (93.3%) subjects on rhPTH(1[ndash]84) and 44 (100%) on placebo reported [ge]1 AE. The most common AEs were hypocalcemia, muscle spasm, paresthesia, headache, and nausea. Four (9%) and 10 (11%) subjects in the PBO and rhPTH(1[ndash]84) groups, respectively, experienced serious AEs; only 1 serious AE (hypercalcemia requiring brief hospitalization) in the rhPTH(1[ndash]84) group was considered treatment-related. There were no deaths. Serum anti-parathyroid hormone antibodies, detected in 2 subjects in the rhPTH(1[ndash]84) group, were low titer. No significant changes in mean cardiovascular parameters (blood pressure, heart rate, or QTc interval) or mean renal parameters (serum creatinine or creatinine clearance) were observed in either group.[br]In this trial of rhPTH(1[ndash]84) for the treatment of HypoPARA, subjects were highly compliant. When rhPTH(1[ndash]84) was withdrawn, close monitoring of serum Ca and symptoms of hypocalcemia were required. Symptoms can be managed by the prompt reintroduction of prior Ca and Vit D dosing that previously controlled the hypocalcemia with empiric adjustments if necessary. For the treatment of HypoPARA, rhPTH(1[ndash]84) was generally well tolerated.[br][br]Sources of Research Support: NPS Pharmaceuticals.[br][br]Disclosures: DS: Advisory Group Member, NPS; Investigator, NPS. BC: Advisory Group Member, NPS. JB: Advisory Group Member, NPS; Principal Investigator, NPS. HL: Employee, NPS. Nothing to Disclose: MLB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1683 187 1353 SUN-325 PO06-01 Sunday 1149 2012


1147 ENDO12L_SUN-326 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Diagnostic Performance of Parathyroid Hormone (PTH) Measurement in Fine-Needle Aspiration Biopsies (FNAB) Washings Alicia Algeciras-Schimnich, Michael A Lasho, Clive S Grant, Ravinder J Singh, Stefan K Grebe Mayo Clinic, Rochester, MN; Mayo Clinic, Rochester, MN [bold]Introduction:[/bold] Parathyroid hormone (PTH) measurement of fine-needle aspiration biopsies (FNAB) needle washings can be used to discriminate thyroid- from parathyroid tissue and can facilitate parathyroid adenoma localization prior to surgery. In this study we reviewed the Mayo Clinic experience with PTH analysis in FNAB needle washings.[br][bold]Objectives:[/bold] To establish a diagnostic cutoff for PTH in FNAB needle washings that identifies parathyroid-derived tissue with 100% specificity.[br][bold]Methods:[/bold] We performed a medical record review of Mayo Clinic patients, who had PTH measurements performed on FNAB needle washings during June 2008 through May 2011. The review included clinical presentation, serum PTH levels, ultrasound- and parathyroid sestamibi scans, FNAB procedures, cytology and pathology reports. The Roche Elecsys PTH immunoassay (Indianapolis, IN) was used for PTH measurements. Statistical analysis was performed using JMP software (version 8.0, SAS Inc.).[br][bold]Results:[/bold] A total of 56 specimens from 50 patients were tested during the study timeframe. In 42 cases, there was sufficient follow-up information to classify the biopsied structures unequivocally as parathyroid- (n=19) or non-parathyroid tissue (n=23). ROC curve analysis showed that a 100 pg/mL cutoff value gave 100% diagnostic specificity; diagnostic sensitivity was 74% at this level. Five cases with confirmed parathyroid tissue had values [lt]100 ng/mL; three of these had also non-diagnostic cytology.On cases with parathyroid tissue confirmed histology (n=19), in which cytology and PTH in FNAB washings was ordered simultaneously (n=18), cytology identified 50% of parathyroid-derived cases, whereas PTH identified 79%.[br][bold]Conclusions:[/bold] Measuring of PTH in FNAB washings is useful in cases of non-diagnostic cytology. At a 100% specificity PTH cut-off (to avoid unnecessary surgery) diagnostic sensitivity is better than cytology, but still suboptimal. This might be improved further by more accurate sampling of the biopsy targets.[br][br]Nothing to Disclose: AA-S, MAL, CSG, RJS, SKG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1811 187 1354 SUN-326 PO06-01 Sunday 1150 2012


1148 ENDO12L_SUN-327 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Familial Pseudohypoparathyroidism Type Ib (PHP-Ib): Clinical Management and Genetic Counseling of a Portuguese Family Maria Lurdes Godinho de Matos, Rute Tomaz, Rita Domingues, Branca Cavaco Hospital de Curry Cabral, Lisbon, Portugal; Instituto Portugues de Oncologia de Lisboa, Lisbon, Portugal [bold]Introduction:[/bold] Familial PHP-Ib is an autosomal dominant disease characterized by hypocalcaemia and hyperphosphataemia due to renal parathyroid hormone resistance.[br]PHP-Ib is caused by an imprinting disruption of [italic]GNAS[/italic] (20q13.3), resulting in tissue–specific silencing of G-protein alpha-subunit. In familial PHP-Ib, maternally inherited 3 kb [italic]STX16[/italic] deletions are associated with a regional [italic]GNAS[/italic] methylation defect.[br]The severity of PTH resistance varies significantly from patient to patient, within the same family.[br][bold]Aims:[/bold] Recently, we identified 8 members of a Portuguese family PHP-Ib, with the 3 kb [italic]STX16[/italic] deletion associated to a [italic]GNAS[/italic] methylation defect.[br]We extended the genetic screening to remaining members in order to improve clinical management and genetic counseling in this family.[br][bold]Methods:[/bold] Twenty two members of a Portuguese family were studied. Clinical presentation and hormonal assays (calcium, phosphorus, parathormone, thyroid FT) were performed. Genetic alterations were assessed.[br]Genomic DNA of each family member was screened for the 3 kb deletion at the STX16, by long range PCR amplification.[br][bold]Results:[/bold] Clinical and genetic studies identified 5 symptomatic PHP-Ib mutation carriers (3 females and 2 males, one with hypothyroidism), 4 asymptomatic mutation carriers (4 males) and 13 unaffected non-carriers (6 females and 7 males).[br]All 5 symptomatic PHP-Ib presented clinical manifestations of acute (e.g. tetanic crisis or seizures) or chronic hypocalcaemia (e.g. ectopic calcifications); all patients had hypocalcaemia, hyperphosphataemia and elevated levels of parathormone in serum. These patients have been treated with oral calcium and vitamin D.[br]Two asymptomatic PHP-Ib mutation carriers (2 males) had elevated levels of PTH with normal calcium and phosphate.[br][bold]Discussion:[/bold] In this study, we have identified a Portuguese family affected with PHP-Ib segregating a 3 kb deletion in the [italic]STX16[/italic] gene maternally, which was found to be associated with loss of methylation at [italic]GNAS[/italic] 1A DMR. Females can transmit the mutant allele to next generation, and the offspring who inherit the deletion will develop the disease, independently of their sex. Males can transmit the mutant allele to next generation but the methylation defect and disease will not occur in the offspring, independently of their sex. In both cases, all carriers will continue to transmit the deletion to future generations with a 50% probability.[br]Routine surveillance of the asymptomatic carriers will be important to prevent future manifestation of disease and hypocalcaemia-related complications. Asymptomatic carriers with elevated PTH, normal calcium and phosphate levels should also be treated in order to normalize PTH levels and to prevent hyperparathyroid bone disease.[br]It will be important to extend the genetic screening to future generations, as it may improve clinical management and genetic counseling in this family.[br][br]Bastepe M, Juppner:GNAS locus and Pseudohypoparathyroidism. Hormone research 2005;63:65-74.[br][br]Nothing to Disclose: MLGdM, RT, RD, BC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1117 187 1355 SUN-327 PO06-01 Sunday 1151 2012


1149 ENDO12L_SUN-328 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Association between Vitamin D Deficiency and Postoperative Normocalcemic Hyperparathyroidism Following Parathyroidectomy for Primary Hyperparathyroidism Daphne D Dadzie, Stanley Z Trooskin, Xiangbing Wang UMDNJ- Robert Wood Johnson Medical School, New Brunswick, NJ; UMDNJ- Robert Wood Johnson Medical School, New Brunswick, NJ [bold]Introduction[/bold]: Primary hyperparathyroidism is the most common cause of hypercalcemia with a prevalence of 1 case per 1,000 people in the United States. The estimated prevalence of vitamin D deficiency (25-hydroxyvitamin D level [lt]20 ng/ml) is estimated to be 41.6% according to the National Health and Nutrition Examination Survey from 2005 to 2006. Minimally invasive parathyroidectomy has become an effective form of treatment for primary hyperparathyroidism with reported cure rates of 95-98%, representing patients with postoperative normocalcemia. There is a subset of patients within this cured group that have been noted to have normocalcemia with persistently elevated PTH levels. The predictive characteristics of postoperative normocalcemic primary hyperparathyroidism (PNPH) are still unknown, but it has been suggested that it may be due to vitamin D deficiency.[br][bold]Methods[/bold]: A sample of 258 charts of primary hyperparathyroidism patients who underwent minimally invasive parathyroidectomy from 2004 to 2010 at Robert Wood Johnson University Hospital was reviewed retrospectively. Calcium, PTH, 25-hydroxyvitamin D serum levels were analyzed preoperatively and postoperatively at 1 month, 3 months, 6 months and 12 months. For this study, [ldquo]cured[rdquo] patients refer to those with calcium levels [lt]10.5 mg/dl and PTH [lt]65 pg/ml, PNPH patients are those with calcium [lt]10.5 mg/dl but PTH level [gt]65 pg/ml, and recurrent or persistent hyperparathyroidism patients are those with calcium [gt]10.5 mg/dl and PTH [gt]65 pg/ml. Comparisons were analyzed with chi-square and t-test.[br][bold]Results[/bold]: Of the 258 patients, 234 (91%) were considered cured and 16 (6%) developed PNPH. The preoperative vitamin D levels were 24.1 ng/ml [plusmn] 15.6 vs 29.2 ng/ml [plusmn] 16.8 (mean and SD) in the PNPH and cured groups respectively. The postoperative vitamin D levels for the PNPH and cured groups were 26.6 ng/ml [plusmn] 8.6 vs 31.9 ng/ml [plusmn] 10.4 at 1 month, 39.4 ng/ml [plusmn] 14.6 vs 35.5 ng/ml [plusmn] 11.0 at 3 months, 39.4 ng/ml [plusmn] 18.1 vs 36.7 ng/ml [plusmn] 16.0 at 6 months, and 39.0 ng/ml [plusmn] 25.5 vs 39.7 ng/ml [plusmn] 15.9 at 12 months. There was no statistical difference between the two groups.[br][bold]Conclusions[/bold]: This study did not show that vitamin D deficiency is the cause of the PNPH nor did it show any statistical difference between the level of vitamin D between the cured and PNPH groups. The predictive characteristics of PNPH are still unclear and it is unknown if vitamin D supplementation preoperatively would reduce the incidence of this condition.[br][br]Nothing to Disclose: DDD, SZT, XW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 639 187 1356 SUN-328 PO06-01 Sunday 1152 2012


1150 ENDO12L_SUN-329 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Preoperative Predictive Factors for Parathyroid Carcinoma in Patients with Primary Hyperparathyroidism Won Sang Yoo, Hyung Jin Choi, Jae Hyun Bae, Jung Hee Kim, Sang Wan Kim, Seong Yeon Kim Seoul National University College of Medicine, Seoul, Korea; Chungbuk National University Hospital, Cheongju Si, Korea Objective: To review the clinical characteristics of parathyroid carcinoma (PC) and to evaluate potential preoperative predictive factors for PC in patients with primary hyperparathyroidism (PHPT).[br]Research Design and Methods: A retrospective review of electronic medical records of 194 patients with pathologically confirmed PHPT in Seoul National University[apos]s affiliated teaching hospitals (Seoul National University Hospital, Metropolitan Government Borame Medical Center, and Seoul National University Bundang Hospital) from January 2000 to March 2011.[br]Results: A total of 194 patients with PHPT, whose etiology was confirmed by a surgically proven pathology, were included in the analysis. Adenoma was diagnosed in 171 patients, hyperplasia in 12, and carcinoma in 11. Mean age at the time of diagnosis was 60 years (range, 46[ndash]74), 48 years (35[ndash]61), and 54 years (40[ndash]68), respectively. There was no significant difference in gender distribution and presenting symptoms according to etiology.[br]Several biochemical measures were higher in patients with PC than in patients with benign disease, including serum total calcium (13.1 [plusmn] 3.4 vs. 11.6 [plusmn] 1.2 mg/dL, p[lt]0.001), intact parathyroid hormone (1152[plusmn] 897 vs. 292[plusmn] 783pg/mL, p[lt]0.01), creatinine (1.4 [plusmn] 0.7 vs. 1.0 [plusmn] 0.4 mg/dL, p[lt]0.001), phosphorous (3.2 [plusmn] 2.0 vs. 2.6 [plusmn] 0.6 mg/dL, p[lt]0.05), and alkaline phosphatase (ALP) (678[plusmn] 560 vs. 137 [plusmn] 329IU/L, p[lt]0.001). Tumors were larger in PC than in benign disease (3.8 [plusmn] 1.6 vs. 1.7 [plusmn] 0.8 cm, p[lt]0.001). Multivariate analysis revealed that serum ALP levels (p[lt]0.05) and tumor size were associated with PC (p[lt]0.001). Tumor size and serum ALP were evaluated as preoperative predictive factors for PC using ROC analyses: a tumor size of 3.0 cm (sensitivity 90.9%, specificity 92.1%) and serum ALP level of 285 IU/L (83.3%, 97.0%) had predictive value for the diagnosis of PC in patients with PHPT.[br]Conclusion: The clinical presentation of patients with PC did not differ significantly from PHPT due to other causes. In patients with PHPT, elevated serum ALP and a large parathyroid mass at the time of diagnosis can be helpful to predict PC.[br][br]Nothing to Disclose: WSY, HJC, JHB, JHK, SWK, SYK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 806 187 1357 SUN-329 PO06-01 Sunday 1153 2012


1151 ENDO12L_SUN-330 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Calcium-Sensing Receptor Polymorphisms Influence on Renal Calcium Handling and Nephrolithiasis in Patients with Primary Hyperparathyroidism Ekaterina A Pigarova, Liudmila Ya Rozhinskaya, Anatoly N Tiulpakov, Svetlana S Mirnaya, Anna V Belyaeva, Natalia G Mokrysheva Endocrinology Research Centre, Moscow, Russian Federation; Endocrinology Research Centre, Moscow, Russian Federation Our aim was to characterize the influence of calcium-sensing receptor (CaSR) polymorphisms on blood and urine biochemical parameters, PTH and nephrolitiasis status in patients with primary hyperparathyroidism (PH) in order to assess its predictive value for nephrolithiasis development in asymptomatic and mild forms of PH.[br]Our study included 135 patients with PH (68M/72F, median age 59), which underwent genetic testing for A986S, R990G and Q1011E polymorphisms of CaSR, evaluation of blood Ca, P, creatinine, PTH, 24h urine levels of Ca, P, and US and medical history data on renal stones. Patients were divided into 2 groups: without nephrolithialis (N-; 69 pts) and with nephrolithiasis (N+; 66 pts).[br]The most common polymorphisms have been CaSR 986S (18%) and 990G (11%), whereas 1011E was seen rarely (5%). There were no differences in the prevalence of polymorphisms between the study groups: for A986S - p = 0,19, R990G - p = 0,67, Q1011E - p = 0.99 (due to the low frequency of homozygous mutations were combined with heterozygotes).[br]Between groups N+ and N-, there were no differences in age, sex ratio, and biochemical serum and urine parameters. In N+ there were higher values of PTH (224,0 [140,0; 577,0] vs. 159,0 [111,0; 270,7], p = 0,013), blood osmolality (0,293 [0,287; 0,305] vs. 0,287 [0,278; 0,297], p = 0,019), and 24h urine osmolality (0,569 [0,421; 0,642] vs. 0,466 [0,355; 0,499], p = 0,034; for those with GFR[gt] 60 ml/min). Polymorphism 986S, in contrast to the wild gene was characterized by loss of the relationship between the level of PTH and the levels of Ca and P (986AA: total Ca and PTH [R = 0,50, p = 0,000026]; ionized Ca and PTH [R = 0,58, p = 0,0000001]; P and PTH [R = - 0,45, p = 0,00025]; 986AS +986 SS total Ca and PTH [R = 0,20, p = 0.28], ionized Ca and PTH [R = 0,11, p = 0,56]; P and PTH [R = - 0,13, p = 0,52], which may indicate the inactivating nature of this polymorphism. Assessing the effect of other studied CASR polymorphisms on blood and urine tests showed no significant differences (Mann-Whitney U-test, p[gt] 0.05 for all parameters).[br]Thus, the incidence of nephrolithiasis in PH is not associated with the presence of alternative alleles of the CaSR gene, so they can not be used to estimate the likelihood of nephrolithiasis in patients with PH. The only significant difference between patients without nephrolithiasis and nephrolithiasis was the increase of blood and urine osmolality, that reflects a greater degree of dehydration.[br][br]Sources of Research Support: The work was supported by grant of President of Russian Federation MK-2852.2010.7.[br][br]Nothing to Disclose: EAP, LYR, ANT, SSM, AVB, NGM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1087 187 1358 SUN-330 PO06-01 Sunday 1154 2012


1152 ENDO12L_SUN-331 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Systemic Inflammation in Primary Hyperparathyroidism: Influences on Plasma Vitamin B6 Levels and Interferon-[gamma] Mediated Immune Activation Monika HE Christensen, Eva KR Pedersen, Yngve Nordbo, Jan Erik Varhaug, Ottar K Nygaard, Gunnar Mellgren, Ernst A Lien University of Bergen, Bergen, Norway; Haukeland University Hospital, Bergen, Norway; Haukeland University Hospital, Bergen, Norway; Haukeland University Hospital, Bergen, Norway; University of Bergen, Bergen, Norway Primary hyperparathyroidism (PHPT) has been linked to low-grade inflammation and enhanced risk of cardiovascular diseases (CVD). The aim of the study was to investigate systemic markers of interferon-[gamma] (IFN-[gamma]) mediated immune activation, such as neopterin, the kynurenine to tryptophan ratio (KTR), kynurenine pathway metabolites, as well as related B6 vitamers in patients with PHPT. We conducted a cross-sectional study on 57 patients with PHPT compared to a control group of 20 healthy blood donors. PHPT patients cured after parathyroidectomy were followed for six months. 43 patients participated in the longitudinal study where blood samples were taken at inclusion and one, three and six months after surgery. Plasma concentrations of the B6 vitamers, pyridoxal 5[apos]-phosphate (PLP) and pyridoxal, were significantly lower in the patient group compared to the healthy controls (p=0.007 and p=0.013, respectively). The kynurenine pathway of tryptophan was influenced with lower plasma concentrations of tryptophan, while both kynurenine and KTR were higher in the patients than in the controls (p=0.007, p=0.032 and p=0.015, respectively). Neopterin was not significantly different between patients and controls at inclusion. During six months after operation, PLP and anthranilic acid increased significantly (both p-values [lt]0.001), whereas neopterin decreased (p=0.018). This study reveals altered level of vitamin B6 and KTR in PHPT patients, which both may reflect systemic inflammation. Some but not all of these abnormalities were corrected 6 months after parathyroidectomy. Low plasma vitamin B6 and elevated KTR are associated with increased risk of CVD, and should be evaluated as CVD risk factors in PHPT.[br][br]Nothing to Disclose: MHEC, EKRP, YN, JEV, OKN, GM, EAL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1902 187 1359 SUN-331 PO06-01 Sunday 1155 2012


1153 ENDO12L_SUN-332 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Recurrent and Persistent Hyperparathyroidism after Parathyroidectomy with a Curative Intent: A 10 Years Review Study Ana Maia Silva, Marta Almeida Ferreira, Jorge Dores, Rui Carvalho, Isabel Palma, Claudia Amaral, Helena Ramos, Helena Cardoso, Conceicao Bacelar, Andre Carvalho, Claudia Freitas, Joana Vilaverde, Jorge Polonia, Fatima Borges Santo Antonio Hospital, Oporto, Portugal; Santo Antonio Hospital, Oporto, Portugal [bold][italic]Background:[/italic][/bold] In experienced centers, most cases of primary hyperparathyroidism (PHPT) will be cured after the first surgery, but recurrence and persistence of hyperparathyroidism can be a challenge to physicians.[br][bold][italic]Aim: [/italic][/bold]To describe cases of recurrent and persistent PHPT and to evaluate the main risk factors associated to it.[br][bold][italic]Methodology:[/italic][/bold] Retrospective analysis of clinical files of patients hospitalized in Santo Antonio Hospital between January/2000 and July/2011 with ICD-9 diagnosis of [ldquo]Hyperparathyroidism[rdquo] and [ldquo]Total Parathyroidectomy[rdquo] or [ldquo]Other Parathyroidectomy[rdquo]. Descriptive analysis of recurrent and persistent PHPT (assumed to be the elevation of parathormone and calcium after normalization and persistent elevated parathormone and calcium 6 months after surgery, respectively) and comparative analysis between them and all cases of cure at 6 months after surgery. T-Student and Fisher[apos]s exact tests were used for statistical analysis.[br][bold][italic]Results:[/italic][/bold] There were 82 operated patients, mainly women (81.7%), with mean age of 59.1[plusmn]14.1 years (27-88) at surgery, with mean follow-up time of 3.3[plusmn]2.9 years (0.1-10.3).[br]The cure rate for PHPT was 84.2% and the recurrent and persistent PHPT rates were 2.4% and 13.4%, respectively.[br]The 2 patients with recurrent PHPT had also MEN1 mutation and were both submitted to a second surgery.[br]Seven of the 11 patients with persistent PHPT were already reoperated and the other four are waiting for surgery; 2 were positive to MEN1 mutation. Only patients MEN1 mutation positive had more than 2 glands parathyroidectomy. There was one case of an ectopic unaccesseble parathyroid. On histological examination, there were 3 cases of normal resected parathyroid glands, 2 cases of parathyroid hyperplasia and one case of no identified parathyroid glands on examination. Preoperative serum parathormone and calcium levels were significantly higher in recurrent and persistent PHPT compared to cured cases of PHPT (572.9 [italic]versus [/italic]251.5 pg/ml and 2.9 [italic]versus[/italic] 2.7 mmol/l, respectively, p[lt]0,05).[br][bold][italic]Discussion[/italic][/bold]: In this study, the incidence of recurrent and persistent PHPT was similar to literature (between 5% and 30%, depending on surgeon[apos]s experience). The risk factors identified were higher preoperative serum parathormone and calcium levels. In some cases adverse features like hyperplasia, ectopic parathyroid gland and MEN1 mutation were also identified. The authors point to the importance of identifying those factors possibly associated with a worse prognosis of PHPT.[br][br]Nothing to Disclose: AMS, MAF, JD, RC, IP, CA, HR, HC, CB, AC, CF, JV, JP, FB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 58 187 1360 SUN-332 PO06-01 Sunday 1156 2012


1154 ENDO12L_SUN-333 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Coexistence of Primary Hyperparathyroidism and Vitamin D Deficiency in a Predominantly African American Patient Population Gauri Dhir, Sajad Salehi, Elias S Siraj Temple University School of Medicine, Philadelphia, PA [bold][italic]Background:[/bold][/italic] Several studies and observations have shown that vitamin D deficiency is more prevalent in subjects with primary hyperparathyroidism (PHPT) than the general population. While some of those data are from United States, African Americans were not adequately represented in most studies.[br][bold][italic]Objective: [/italic][/bold]To determine and characterize the association of vitamin D deficiency and PHPT in a predominantly African American patient population at an endocrine outpatient clinic of a University Hospital.[br][bold][italic]Methods:[/italic][/bold] We conducted a 5 year (2007-2011) retrospective chart review of 70 patients that presented with suspicion of Primary Hyperparathyroidism to the Endocrine Clinic of Temple University Hospital.[br][bold][italic]Results:[/italic][/bold] Of the 70 patients, 49 had a definitive diagnosis of PHPT and were included in the analysis. The mean age was 64 years, 85% were females and 50% were African Americans.[br]When looking at the relationship between PTH levels and calcium levels, 66% (33/49) had both high PTH ([gt]60 pg/mL) and high Ca ([gt]10 mg/dL), 28% (14/49) had high PTH and normal calcium while 4% (2/49) had normal PTH and high calcium. Mean 25 (OH) VD levels were not different in those 3 groups.[br]About 80% (37/46) of subjects had vitamin D insufficiency defined as 25 (OH) VD levels of [lt] 30 ng/mL while 48% (22/46) had vitamin D deficiency defined as levels of [lt] 20 ng/mL. Mean serum PTH and Ca levels, 24 hours urine calcium levels and DEXA scan T-scores were not different whether VD levels were low or not.[br]About 40% (19/49) of our subjects had osteoporosis and 18% (9/49) had history of kidney stones. The 24 hours urine calcium levels correlated positively with serum calcium levels as well as with 1,25 (OH) VD levels (P= 0.002 and 0.023 respectively) but not with 25 (OH) VD or PTH levels. Calcium levels positively correlated with PTH levels (P=0.006).[br][bold][italic]Conclusion:[/italic][/bold] Our study documents the relatively high prevalence of vitamin D deficiency/insufficiency in a predominantly African American population of PHPT. Unlike some previous studies and observations, our study did not demonstrate differences in clinical and biochemical features whether patients with PHPT had low VD levels or not. We believe that the relatively high prevalence of inadequate VD status in our population of PHPT calls for more studies to better understand its impact and find out the best therapeutic approaches.[br][br]Nothing to Disclose: GD, SS, ESS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 94 187 1361 SUN-333 PO06-01 Sunday 1157 2012


1155 ENDO12L_SUN-334 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Severe Elevation in Intact PTH Secondary to Hungry Bone Syndrome after Resection of Type [quot]F[quot] Atypical Parathyroid Adenoma Muhammad Houri Sparks Health System, Fort Smith, AR 56 year old female presented in October of 2010 with severe hypercalcemia with a calcium level of 20 mg/dL and intact PTH of more than 3000 pg/ml and she also had chronic renal insufficiency with worsening renal function and severe lower extremity weakness, which made her bedridden. Hypercalcemia was treated with hydration, calcitonin, a dose of Zometa and eventually one session of hemodialysis.[br]Because of the clinical presentation suspicion for parathyroid carcinoma was raised. Parathyroid scan revealed uptake in the mediastinum. Ultrasound revealed two thyroid nodules, both in the posterior aspect of right thyroid lobe. Because of the clinical suspicion for parathyroid carcinoma and the fact that her uptake was in the mediastinum, the patient was referred to a tertiary care center.[br]The findings at surgical exploration were consistent with type F (fallen to mediastinum) parathyroid adenoma with atypia. After surgery her PTH dropped by more than 90%. However, when she came back for follow up in my office, her PTH was up to 1000 pg/ml with low potassium, low magnesium, and hypocalcemia, all consistent with hungry bone syndrome. The patient was started on calcium, magnesium, and potassium supplement along with Rocaltrol and her calcium level improved and her intact PTH level has decreased slowly but steadily since then. Her intact PTH is still elevated at more than 300 pg/mL. It was as high as 1000 mcg/mL when she returned back from her surgery. Fortunately, her calcium has remained normal since her surgery.[br]The presentation with severe hypercalcemia, renal insufficency osteoprosis and elevated bone turnover markers was suspicious for parathyroid carcinoma. Because of the diagnosis of parathyroid carcinoma could not be easily made on pathology, and may present with recurrence of hypercalcemia after successfol resection of [quot] an adenoma [quot] the patient will need continued follow up. Another possibility for her presentation may be that her severe hypercalcemia was made worse by the fact that she was immobile for almost two months prior to presentation.[br][br]Nothing to Disclose: MH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2344 187 1362 SUN-334 PO06-01 Sunday 1158 2012


1156 ENDO12L_SUN-335 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Are PTH Normal Ranges in the Elderly Method Dependent? Preserved Renal Function Estimation by Using Different Indexes Andrea Kozak, Ana M Sequera, Viviana Mesch, Patricia Otero, Paula Esteban, Graciela Astarita, Monica Saavedra, Isabel Teres, Patricia Pagano, Maria J Iparraguirre, Marta Torres, Mirta Gurfinkiel Argentine Society of Endocrinology and Metabolism (SAEM), Buenos Aires, Argentina The aim of the present collaborative work was to re-evaluate normal ranges of parathyroid hormone in the elderly in order to include subjects over 80 years old, as life expectation has risen in the last years. Material and Methods: We investigated retrospectively data from patients attending to the various hospitals integrating the Biochemistry Department. We selected a population of patients with the following inclusion criteria: preserved renal function, non-diabetic, normal levels of calcium, both in serum and urine, Vitamin D [ge] 30 ng/ml, euthyroid, no treatment with corticoids, anti resorptives or estrogens, and normal arterial tension. They were divided into 3 groups according to age: Group A (50-60 years), B (61-70 years) and C (over 71 years). Methods for PTH included in the comparison were ICMA Immulite[trade] Siemens, ECLIA Cobas[trade] Roche and ICMA Architect[trade] Abbott. Two different formulas were used to estimate renal function, MDRD and CDK-EPI. The number of subjects included in each age group per method was ICMA Immulite n: 145 (Group A: 54, Group B: 39, Group C: 52); ECLIA Cobas n: 93 (Group A: 40, Group B: 30 and Group C: 23) and ICMA Architect n: 168 (Group A: 54, Group B: 52 and Group C: 62). Statistical analysis included Anova, Kruskall-Wallis, Bland-Altman for methods[acute] differences, and Spearman Ranks correlation for evaluation of relationship between PTH and age. Results: The results showed no significant differences between age groups assayed by the different methods: Group A (50-60 years), PTH ICMA Immulite, median: 48.5 pg/ml vs. PTH ECLIA Cobas, median: 43.8 pg/ml vs. PTH ICMA Architect, median: 46.6 pg/ml; Group B (61-70 years), PTH ICMA Immulite, median: 57.4 pg/ml vs. PTH ECLIA Cobas, median: 50.5 pg/m vs. PTH ICMA Architect, median: 53.1 pg/ml; Group C (71-89 years), PTH ICMA Immulite, median: 56.6 pg/ml vs. PTH ECLIA Cobas, median: 50.4 pg/ml vs. PTH ICMA Architect, median: 50.8 pg/ml. Also no significant differences were found between age groups assayed by the same method, and no association was found between age and PTH level. Conclusions: PTH levels are similar between age groups evaluated, making it possible to use the adult reference range for the elderly. The condition for use is that renal function is preserved (evaluated with creatinine clearance and formulae for glomerular filtration rate) and that adequate Vitamin D levels are present.[br][br]Nothing to Disclose: AK, AMS, VM, PO, PE, GA, MS, IT, PP, MJI, MT, MG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1302 187 1363 SUN-335 PO06-01 Sunday 1159 2012


1157 ENDO12L_SUN-336 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Epidemiology of Primary Hyperparathyroidism and Its Nonclassical Manisfestations in the City of Recife, Brazil Catia Sousa Eufrazino, Narriane Chaves Holanda, Paula Aragao Prazeres, Francisco Alfredo Bandeira University of Pernambuco Medical School, Recife, Brazil Primary hyperparathyroidism (PHPT) is the main cause of hypercalcemia in patients in ambulatory care1. Before the introduction of routine measurements of serum calcium in ambulatory services, PHTP was symptomatic with classic bone disease, known as osteitis fibrosa cystica, nephrolithiasis and acute neuropsychiatric syndrome with severe hypercalcaemia. In most series, usually PHPT is asymptomatic or presents few unspecific symptoms2,3. The aim of this study was to determine the prevalence of PHPT in patients attending reference centers (public and private). Materials and Methods: The diagnostic criteria for PHPT were as follows: elevated serum calcium in two occasions plus serum PTH above 75o centile for the reference population (57pg/ml). The period prevalence was calculated as the number of existing cases for 9 months divided by population and expressed as a percentage of every 1000 population; 95% confidence intervals (CIs) were estimated assuming a Binomial distribution. Results: From 4.207 patients we found a prevalence of PHTP of 0,78 (95% CI 0,52-1,04) of which 81.8% were asymptomatic and 18.2% symptomatic. The female/male ratio was 7.2:1, and 89.7% of these women were postmenopausal. Mean age was 61.12 [plusmn] 15.73 years, serum calcium 10.63 [plusmn] 1.33mg/dl and serum PTH 182.48 [plusmn] 326.51 pg/ml. At the public center the mean serum calcium level was 11.05 [plusmn] 1.36mg/dl and serum PTH was 198.17 [plusmn] 400,69 pg/ml, while at the private center they were 9.88 [plusmn] 0.92mg/dl (p=0.01) and 155.03 [plusmn] 130.07 pg/ml (p=0.72), respectively. Osteitis fibrosa cystic was present in 6.1% (2/33); nephrolithiasis in 18.2% (6/33) and acute neuropsychiatric syndrome in 3% (1/33). With regard to non-classical manifestations: 51.5% (17/33) had fatigue and 39.3% (13/33) muscle weakness. Moreover, 63.6% (21/33) had hypertension; 33% (9/33) type 2 diabetes mellitus; 18.2% (6/33) depression; 6.1% (2/33) peptic ulcer and MEN-1. Conclusion: We found a high prevalence of PHTP at endocrine reference centers. The presence of higher serum calcium and PTH levels in the public service suggests a later medical assessment.[br][br](1) Jorde R et al., J Clin Epidemiol 2000; 53:1164-9. (2) Dalemo S et al., Scandinavian Journal of Primary Health Care 2006; 24:160-165. (3) Bilezikian JP et al., N Engl J Med 2004; 350:1746-1751.[br][br]Nothing to Disclose: CSE, NCH, PAP, FAB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1119 187 1364 SUN-336 PO06-01 Sunday 1160 2012


1158 ENDO12L_SUN-337 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Parathyroid Tumors Express an Embryonic Stem Cell Gene Signature Valentina Vaira, Chiara Verdelli, Irene Forno, Francesca Elli, Vito Guarnieri, Stefano Ferrero, Alfredo Scillitani, Elena Costa, Silvano Bosari, Anna Spada, Sabrina Corbetta Fondazione IRCCS C[agrave] Granda Ospedale Maggiore Policlinico, Milan, Italy; IRCCS Policlinico San Donato, San Donato Milanese, Italy; University of Milan, Fondazione IRCCS C[agrave] Granda Ospedale Maggiore Policlinico, Milan, Italy; University of Milan, Fondazione IRCCS C[agrave] Granda Ospedale Maggiore Policlinico, Milan, Italy; IRCCS Hospital Casa Sollievo della Sofferenza, San Giovanni Rotondo, Italy; IRCCS Hospital Casa Sollievo della Sofferenza, San Giovanni Rotondo, Italy; University of Milan, IRCCS Policlinico San Donato, San Donato Milanese, Italy In our previous investigation on microRNAs expression pattern in parathyroid carcinomas (Ca) by Taqman low-density array profiling, we detected the overexpression of microRNAs belonging to C19MC (MIR517A, MIR517B, MIR517C, MIR518A, MIR518C, MIR518E, MIR518F, MIR520H, MIR522), the largest human cluster on chromosome 19q13.41 with respect to normal parathyroid glands. In the present study, the analysis of the expression of selected C19MC and the closely distal MIR371-3 clusters microRNAs (MIR512-3p, MIR517C, MIR520H and MIR372) was extended to 11 Ca, 24 adenomas (Ad) and 6 normal glands. The four microRNAs was expressed in 11% of parathyroid neoplasia. Almost all Ca showed elevated MIR517C levels compared to normal glands, while MIR517C overexpression occurred only in two Ad. MIR372 was detected in 45.5% of Ca and in 37.5% of Ad. By copy number variation analysis, C19MC amplification was identified in most Ca extending distal to the MIR371-3 cluster in almost all amplified samples. Conversely, C19MC amplicon was detected in a small subset of Ad extending distal to MIR371-3 in one sample. C19MC promoter was hypomethylated in 38% of the samples with no difference in frequency between Ad and Ca. Few tumours did not show C19MC amplicon nor hypomethylation.[br]Since the C19MC as well as the MIR371-3 clusters have been linked with the signature characteristic for human embryonic stem cells, we investigated genes of the transcriptional regulatory network governing stem cells pluripotency and self-renewal. By immunohistochemistry, cells with a positive nuclear staining for NANOG were more abundant in Ca (30-70%, n=8) compared to normal glands ([lt]10%, n=3). Most Ads (n=11) expressed NANOG at very low levels (1-7%), while two samples only, characterized by mild hypercalcemia and very mild PTH hypersecretion, showed NANOG expression levels of 25% and 40%, respectively. In parathyroid tumors, [italic]NANOG[/italic] expression was associated with the expression of the core genes [italic]OCT4 [/italic]and[italic] SOX2.[/italic] A subset of [italic]NANOG[/italic]-expressing tumors also expressed the[italic] KLF4 [/italic]gene. Moreover, [italic]zfp42/REX1[/italic] gene expression was lost in some tumours, as described in other human cancers. [italic]DKK1[/italic] gene expression was undetectable in all samples. These data identified a genetic embryonic stem cell signature in parathyroid tumours that seems to correlate with more aggressive tumoral features.[br][br]Nothing to Disclose: VV, CV, IF, FE, VG, SF, AS, EC, SB, AS, SC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1524 187 1365 SUN-337 PO06-01 Sunday 1161 2012


1159 ENDO12L_SUN-338 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Primary Hyperparathyroidism [mdash] Initial Experience of Atypical Presentations in a Tertiary Care Center of India Sadish Kumar Kamalanathan, Karthik Balachandran, Girish Parthan, Abdoul Hamide, Amit Goel, Dhanapathi Halanaik, Jayaprakash Sahoo Jawaharlal Institute of Postgraduate Medical Education [amp] Research (JIPMER ), Puducherry, India; Jawaharlal Institute of Postgraduate Medical Education [amp] Research (JIPMER ), Puducherry, India; Jawaharlal Institute of Postgraduate Medical Education [amp] Research (JIPMER ), Puducherry, India; PIMS, Puducherry, India BACKGROUND: Symptomatic presentation of primary hyperparathyroidism (PHPT) is the norm rather than exception in India unlike in the West. AIM: To report and analyze retrospectively seven cases of PHPT who presented to JIPMER in the past two years. METHODS: All patients with biochemical profile consistent with PHPT were confirmed and localized with ultrasound and MIBI scan. Renal and bone nutrition status were evaluated. Metabolic status was optimized and parathyroidectomy done in all symptomatic cases. RESULTS: All patients except one were less than 50 years of age. Similarly all except one had a symptomatic presentation. Three patients presented with acute pancreatitis. Another two had neuromuscular weakness associated with severe myopathy. One presented with bilateral maxillary masses. All except the asymptomatic patient had grossly elevated PTH and alkaline phosphatase levels.Renal stones were present in three and osteoporosis was present in all of six evaluated. Three had osteitis fibrosa cystica.Vitamin D deficiency and insufficiency was documented in two patients each. Parathyroidectomy of all isolated adenomas cured all but one patient.The postoperative biopsies of all cured patients revealed parathyroid adenoma while that of uncured patient showed parathyroid carcinoma. Two developed hungry bone syndrome postoperatively.DISCUSSION:The two patients who presented with neuromuscular weakness and severe myopathy had severe 25(OH)D deficiency associated with PHPT. Calcium which was normal in two patients rose following resolution of pancreatitis in one and with cholecalciferol supplementation in another. Hypophosphatemia was constant feature in all seven patients. The asymptomatic patient with probable MEN 1 syndrome has unilateral parathyroid adenoma and is on observation despite being young and deserving for surgery. The uncured parathyroid carcinoma patient is currently on bisphosphonate therapy. Vitamin D insufficiency/deficiency induced secondary hyperparathyroidism contributes significantly to the symptomatology of PHPT. CONCLUSION: Symptomatic PHPT is still the prevalent form of presentation of PHPT in India. Calcium can be normal in PHPT in the face of concomitant 25(OH)D deficiency or acute pancreatitis. Hypophosphatemia is a consistent feature of PHPT when compared to hypercalcemia.[br][br]Nothing to Disclose: SKK, KB, GP, AH, AG, DH, JS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 290 187 1366 SUN-338 PO06-01 Sunday 1162 2012


1160 ENDO12L_SUN-339 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Combined Accuracy of Radiologist and Surgeon-Performed Preoperative Imaging Studies in Primary Hyperparathyroidism: Does IOPTH Add Any Benefit? Jennifer H Kuo, Raymon Grogan, Elliot Mitmaker, Lauren Ogawa, Jessica E Gosnell, Orlo H Clark, Quan Y Duh, Wen T Shen University of California, Davis, Sacramento, CA; University of California, San Francisco, CA Background: Surgeons often use preoperative ultrasound along with one other imaging study for preoperative planning in primary hyperparathyroidism (PHPT), and rely upon intraoperative PTH (IOPTH) measurement to confirm cure. The accuracy of combined radiologist and surgeon-performed imaging in predicting single-gland disease in PHPT is unknown. We studied the combined accuracy of radiologist and surgeon-performed imaging in order to determine whether IOPTH adds any benefit in this setting.[br]Method: This was a single-institution review of 910 PHPT patients from 2005 to 2010. Radiologist concordance was defined as concordance of ultrasound and sestimibi as read by attending radiologists. Surgeon concordance was defined as concordance of surgeon-performed ultrasound with at least one of the radiologist-performed studies. Single-gland disease was defined by histological abnormality and biochemical cure (normal calcium and PTH 6 months post-op) after removal of a single gland. A univariate analysis identified risk factors for multiglandular disease.[br]Results: Of the 910 study patients, 671 (73.7%) were women with a mean age of 61+/-12.6 years. Radiologist-performed ultrasound was done in 882 (96.9%) patients, sestamibi in 893 (98.1%), surgeon-performed ultrasound in 468 (51.4%), and IOPTH in 725 (79.7%). Family history and previous exploration were identified as risk factors for multiglandular disease (n=140). The positive predictive values of radiologist concordance were 89.1%, 90% and 83.8% in all patients (AP), low risk patients (LR) and high risk patients (HR) respectively, with an added benefit of 8.0%, 7.5%, and 11.3% with the addition of IOPTH (p[lt]0.05). Surgeon-performed ultrasound was more accurate (AP 93.7%, LR 93.7%, and HR 0.6%), with less benefit gained by IOPTH (AP 1.5%, LR 0.8%, and 4.6%, p[gt]0.05). Surgeon concordance with preoperative radiologist-performed imaging was most accurate (AP 94.8%, LR 95.0%, HR 92.9%) with no extra benefit gained from IOPTH in the AP and LR patient groups (AP -0.1%, LR -0.2%, and HR 1.9%, p[gt]0.05).[br]Conclusion: The addition of a surgeon-performed ultrasound to radiologist-performed imaging studies improves the accuracy of predicting single gland disease in PHPT with imaging studies. In addition, in patients who are low-risk for multiglandular disease, IOPTH may actually decrease the benefit of preoperative imaging studies.[br][br]Nothing to Disclose: JHK, RG, EM, LO, JEG, OHC, QYD, WTS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 130 187 1367 SUN-339 PO06-01 Sunday 1163 2012


1161 ENDO12L_SUN-340 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Identification and Characterization of D410E, a Novel Mutation in the Loop 3 Domain of CaSR, in Autosomal Dominant Hypocalcemia and Therapeutic Approaches with Novel Calcilytics, AXT914: A Series of [italic]Korean Hypopara Registry Study (3)[/italic] So Young Park, Hee-Chang Mun, Arthur D Conigrave, Tae Sik Jung, Suntaek Hong, Young Sil Eom, Ie Byung Park, Sihoon Lee Cheil General Hospital, Kwandong University College of Medicine, Seoul, Korea; University of Sydney, Sydney, Australia; Gyeongsang National University College of Medicine, Jinju, Korea; Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Incheon, Korea; Gachon University School of Medicine, Incheon, Korea [bold]Background:[/bold] Autosomal dominant hypocalcemia (ADH) can be caused by activating mutations of the calcium-sensing receptor [italic](CaSR)[/italic] gene. CaSR plays a pivotal role in the regulation of calcium homeostasis and is abundantly expressed in parathyroid gland, thyroid C cell, and renal tubular system. Activation of CaSR by increased Ca[sup]2+[/sup] inhibits PTH secretion, stimulates calcitonin secretion, and promotes urinary Ca[sup]2+[/sup] excretion, and thereby maintains the Ca[sup]2+[/sup] at the normal level. Herein, we report a novel activating mutation in the [italic]CaSR[/italic] gene in a Korean family with ADH. Meanwhile, antagonist of the CaSR, calcilytics could be a therapeutic option in the treatment of ADH.[br][bold]Method:[/bold] We identified a 55-yr-old woman with mild hypocalcemia (8.0 mg/dL) and hypercalciuria (24-hr u Ca: 441 mg/d) caused by missense mutations of the[italic] CaSR[/italic] gene. She showed low normal PTH level (10.14 pg/mL). We performed mutational analysis of the genes encoding GCMB, PTH and CaSR in her family members. The ability of wild-type and mutant CaSR to activate the MAPK signaling cascade was assessed by examining phosphorylation of ERK1/2 and to increase intacellular Ca[sup]2+[/sup] mobilization was assessed by the Ca[sup]2+[/sup]-sensitive dye fura-2 AM. Blocking of CaSR with calcilytics, AXT914 was also assessed by fura-2 with a variety of concentrations.[br][bold]Result:[/bold] Direct sequencing analysis of the[italic] CaSR[/italic] gene showed that the proband and her daughter possess a T to A transition at nucleotide 1230 resulting in a D410E missense mutation in exon 4 of the [italic]CaSR.[/italic] HEK293 cells were stably transfected with plasmids encoding wild-type or mutant CaSR genes. Escalation of the extracellular Ca[sup]2+[/sup] concentration from 0.5 to 5.0 mM resulted in increased phosphorylation of ERK1/2 and escalation of the extracellular Ca[sup]2+[/sup] concentration from 1.0 to 10 mM resulted in increased intracellular Ca[sup]2+[/sup] detected by fura-2 in mutant CaSR-expressing cell than wild-type-expressing one. These results indicate that D410E mutation of CaSR is associated with ADH in this family. Finally, AXT914 successively blunted the increased intracellular signaling via CaSR.[br][bold]Conclusion:[/bold] We add one more [italic]CaSR[/italic] activating mutation that causes ADH to over 60 activating mutations that have been identified so far. This is of interest because this novel mutation occurred in the loop 3 region of the VFT domain in CaSR where little was known to be important in its function. Further clinical study is needed to validate the effectiveness of calcilytics in the treatment of ADH [italic]in vivo[/italic].[br][br]Sources of Research Support: This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology, Korea [2011-0005621 to S.L.].[br][br]Nothing to Disclose: SYP, H-CM, ADC, TSJ, SH, YSE, IBP, SL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 896 187 1368 SUN-340 PO06-01 Sunday 1164 2012


1162 ENDO12L_SUN-341 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) A Randomized, Double-Blind, Placebo-Controlled Study To Investigate Use of Recombinant Human Parathyroid Hormone (rhPTH[1[ndash]84]) for the Treatment of 134 Adults with Hypoparathyroidism: Baseline Demographics and Subject Characteristics Michael Mannstadt, Tamara Vokes, Peter Lakatos, Hjalmar Lagast Massachusetts General Hospital and Harvard Medical School, Boston, MA; University of Chicago, Chicago, IL; Semmelweis University Medical School, Budapest, Hungary; NPS Pharmaceuticals, Bedminster, NJ Hypoparathyroidism (HypoPARA) is a rare endocrine disorder in which the parathyroid glands are absent or produce insufficient parathyroid hormone (PTH) which causes hypocalcemia, hyperphosphatemia, and hypercalciuria. Therapy for HypoPARA is limited to symptomatic management, often with large amounts of calcium (Ca) and active vitamin (Vit) D metabolites, resulting in possible long-term complications such as end-organ damage.[br]This randomized, double-blind, placebo (PBO)-controlled study assessed the safety and efficacy of recombinant human PTH (rhPTH[1[ndash]84]) in reducing the need for large doses of Ca and active Vit D, while normalizing or maintaining albumin-corrected total serum Ca concentration. Before randomization, subjects underwent a 2[ndash]16 week optimization period when oral Ca and Vit D doses were adjusted to achieve serum Ca concentrations of 8.0[ndash]9.0 mg/dL.[br]A total of 90 and 44 subjects were randomized to rhPTH(1[ndash]84) and PBO groups, respectively. Overall, 77% and 82% were female; 94% and 98% were white; and mean ages were 47.0 and 48.5 years. Subject recruitment was balanced across 29 US and European centers. HypoPARA resulted from surgery in 75.6% of rhPTH(1[ndash]84) and 70.5% of PBO subjects. Other causes included underlying genetic disorder, radiation exposure, or autoimmune and idiopathic disease. Mean [plusmn] SD duration of HypoPARA was 14.1 [plusmn] 11.1 and 11.0 [plusmn] 8.0 years in rhPTH(1[ndash]84) and PBO subjects, respectively.[br]At baseline, the prescribed Ca dosage was [gt]2000 mg/day for 32.2% of rhPTH(1[ndash]84) and 29.5% of PBO subjects. Also, 67.8% of rhPTH(1[ndash]84) and 63.6% of PBO subjects were taking [gt]0.5 [mu]g/day of calcitriol or [gt]1.0 [mu]g/day of alphacalcidol.[br]The respective mean [plusmn] SD baseline serum laboratory values for the rhPTH(1[ndash]84) and PBO groups were similar: Ca, 8.8 [plusmn] 0.8 and 9.0 [plusmn] 0.7 mg/dL, respectively; phosphate, 4.5 [plusmn] 0.3 and 4.6 [plusmn] 0.7 mg/dL; alphacalcidol, 43.3 [plusmn] 18.4 and 44.2 [plusmn] 18.2 ng/mL; calcitriol, 34.3 [plusmn] 20.9 and 32.6 [plusmn] 11.5 pg/mL; and urine Ca, 351.7 and 340.1 mg/24hr. Mean bone mineral density (BMD) was 1.2 vs 1.2 g/cm[sup]2[/sup] for the lumbar spine and 1.1 vs 1.0 g/cm[sup]2[/sup] for total hip, respectively.[br]These baseline data demonstrate that rhPTH(1[ndash]84) and PBO groups had comparable demographic and disease-based characteristics, Ca and active Vit D metabolite doses, and laboratory values. Serum Ca was at the target level (lower end of the normal range), whereas urine Ca was markedly elevated and BMD was increased, as has been previously reported for patients with HypoPARA.[br][br]Sources of Research Support: NPS Pharmaceuticals.[br][br]Disclosures: MM: Advisory Group Member, NPS. TV: Advisory Group Member, NPS. HL: Employee, NPS. Nothing to Disclose: PL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1796 187 1369 SUN-341 PO06-01 Sunday 1165 2012


1163 ENDO12L_SUN-342 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Ultrasound Is Superior to Computed Tomography for Assessment of Nephrocalcinosis in Hypoparathyroidism Alison M Boyce, Thomas A Shawker, Suvimol C Hill, Peter L Choyke, Michael C Hill, Robert James, Nancy A Yovetich, Michael T Collins, Rachel I Gafni National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; George Washington University Hospital, Washington, DC; Rho, Inc, Chapel Hill, NC Nephrocalcinosis (NC) is calcification of the renal parenchyma, which in severe cases may lead to renal insufficiency. Surveillance for NC in hypercalciuric conditions such as hypoparathyroidism (HP) is an important component of management, as NC is asymptomatic and early diagnosis may lead to improved outcomes. Imaging modalities for NC include ultrasonography (US) and computed tomography (CT). Few studies have compared these modalities, and standard practice is not defined. Given the intrinsic limitations of CT in detecting small tissue deposits, we hypothesized that US may be superior for detecting NC. Methods: Concurrent US and CT images were obtained from 22 HP subjects (9-50y) and 7 controls without known risk factors for NC (30-74y). Three copies of each scan were created; each was de-identified and assigned a unique ID#. Two radiologists specializing in US read the 87 US scans and 2 CT radiologists read the 87 CT scans. Thus, within each subject there were 6 replicate US scores (2 radiologists x 3 independent readings) and 6 replicate CT scores, totaling 174 replicate readings per scan type. NC was classified using a 0-3 scale with 0 = no NC and 3 = severe NC. Replicate scores within and between devices (within a subject) were analyzed to determine inter-rater and inter-device agreement via multi-observer Kappa statistics. A generalized linear mixed cumulative logit model (GLMM), with random effects for subject and specialist, tested for differences in scores between the devices. Results: Only moderate agreement was found between US and CT scores, with an inter-device kappa=0.47 (0.22, 0.66) and 60% concordance. Of the 40% discordant pairs, 81% had higher US scores and only 19% had higher CT scores. Eighty (46%) of the 174 replicate US scores were [gt] 0, while only 47 (27%) of the 174 replicate CT scores were [gt] 0. Of the NC cases seen on US and not CT, 45%, 46%, and 9% were grades 1, 2, and 3, respectively. Overall, US scores were higher than CT scores with a cumulative odds ratio of 5.97 (2.60, 13.75), p[lt]0.01. In the 7 control subjects 100% of the US ratings were 0, whereas 95% of the CT ratings were 0. Conclusions: US appears to be superior to CT for detecting mild to moderate NC in patients with HP. This superior sensitivity did not translate into higher readings in control subjects. Given its potential greater sensitivity, lack of radiation, and low cost, US appears to be the preferred modality for the diagnosis and monitoring of NC.[br][br]Sources of Research Support: Intramural Research Program of the NIH, NIDCR and NICHD.[br][br]Nothing to Disclose: AMB, TAS, SCH, PLC, MCH, RJ, NAY, MTC, RIG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 782 187 1370 SUN-342 PO06-01 Sunday 1166 2012


1164 ENDO12L_SUN-343 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Case Study of a Patient with Pseudohypoparathyroidism and Learning Disability Zohreh Shoar, Kimberly B Fuld St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA Background: Pseudohypoparathyroidism (PHP) is a group of disorders with elevated levels of parathyroid hormone (PTH) and biochemical features of hypoparathyroidism, specifically hypocalcemia and hyperphosphatemia. In this condition, the parathyroid glands are normal or hyperplastic and can synthesize and secret PTH but target tissues are unresponsive to its biological effects. The disorders are classified according to different phenotypic and biochemical findings and based on genetic defects in the hormone receptor adenylate cyclase system. PHP-1B is a renal-specific resistance to PTH that lacks the phenotypic features and bone deformities seen in the more classic PHP-1A. Genetic linkage analysis has shown an imprinting defect locus on chromosome 20q13.3 that includes GNAS, a gene that encodes G-protein [alpha]-subunit, a signaling protein in this disorder. Paternal uniparental idiosomy and methylation defects of GNAS, along with loss of maternal GNAS, leads to PTH resistance and mineral dysregulation in the proximal renal tubules in PHP-1B.[br]Clinical Case: A 7-yr old previously healthy male (DB) with learning difficulty and an otherwise negative family history, was brought to ED for a 10 minute tonic-clonic seizure while sleeping. Upon arrival, he was awake, responsive but showed clinical evidence of hypocalcemia with unremarkable physical exam. Initial laboratory assessment showed a calcium level of 5.6 mg/dL, phosphate of 8.7 mg/dL, and ionized calcium of 0.71 MMOL/L and 0.64 MMOL/L in a 2 hour period. DB was started on a high dose calcium infusion to prevent further seizure activity. Further assessment showed elevated PTH levels of 321 pg/mL with the concurrent calcium of 5.5 mg/dL, mild vitamin D-25-OH deficiency of 14.1 ng/mL, normal vitamin D-1-25-OH, TFT, and spot urinary calcium. Renal ultrasound and bone survey were normal. Oral calcium, calcitriol, and ergocalciferol were implemented to normalize the ionized calcium and phosphate while the calcium infusion rate was weaned. He was discharged from the hospital on oral medications in stable condition with follow-up. [br]Conclusion: Our patient is a case of PHP-1B and learning disability. The association between PHP-1B, developmental delay and mental retardation has been described in 3 patients. Our patient is another unique case of learning difficulties in the context of PHP-1B diagnosis. Genetic testing, which has been pursued, will clarify the molecular pathogenesis of the disease.[br][br]Nothing to Disclose: ZS, KBF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1152 187 1371 SUN-343 PO06-01 Sunday 1167 2012


1165 ENDO12L_SUN-344 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) Hypocalcemia: A Reversible Cause of Systolic Heart Failure Nuttha Ungnapatanin, Shema R Ahmad, Jose S Subauste University of Mississippi, Jackson, MS [bold]Introduction:[/bold] Systolic heart failure (SHF) is a common condition which 5,800,000 people in the United States carried this diagnosis in year 2006.[sup]1[/sup] Hypocalcemia is one of reversible etiologies of SHF. Here we describe a case of suspected DiGeorge syndrome who presented with acute decompensated systolic heart failure (ADSHF) and hypocalcemia due to primary hypoparathyroidism.[br][bold]Case description:[/bold] A 45-year old man presented with ADSHF. He has history of tetralogy of Fallot (TOF), s/p surgical repair at age 3. He has not had any follow up and was not on medication. Initially, labs were compatible with primary hypoparathyroidism; calcium level was low (5.5 mg/dL) with inappropriately normal level of intact PTH (15.3 pg/ml), high phosphorus (6.7 mg/dL), low magnesium (1.2 mg/dL) and low 25-OH vitamin D level (11.9ng/ml). Physical findings were low set ears, ocular hypertelorism, Chvostek[apos]s sign, poor dentition and vitiligo. Transthoracic echocardiogram showed severely reduced left ventricular (LV) systolic function with global hypokinesis. Estimated LVEF was 20%. No significant valvular disease was found. Hypocalcemia was initially treated with intravenous calcium gluconate. Upon discharge, his regimen was calcitriol 0.5mcg BID, Ergocalciferol 50,000units weekly and calcium carbonate 1.2gm BID. Four months after, his calcium was normal (9.4 mg/dL) with calcitriol 0.25mcg and calcium carbonate 600mg daily. Repeat transthoracic showed significantly improved LV systolic function with LVEF of 45-50%. With his clinical features and history of TOF, we suspect DiGeorge syndrome.[br][bold]Discussion:[/bold] Calcium has a dominant role in the sequence of myocardial excitation- contraction coupling and myocardial relaxation. Hypocalcemia prolongs phase 2 of the action potential duration and the Q-T interval.[sup]2[/sup] Chronic hypocalcemia can also cause dysrhythmias, decreased myocardial performance, and SHF.[sup]3[/sup] Prevalence of hypocalcemia-induced SHF in adult has not been reported. Tomar M, et al. did a study in India and found that 15 out of 94 (16%) babies with severe LV dysfunction had severe hypocalcemia.[sup]4[/sup] Myocardial dysfunction secondary to hypocalcemia is reversible with calcium repletion. Significantly improved LV function from treatment of hypocalcemia in patient without underlying myocardial disease was first reported in 1985.[sup]5[/sup] Hypocalcemia should be included in the differential diagnosis of patients with SHF and unnecessary diagnostic and therapeutic measures might be avoided.[br][br](1) Lloyd-Jones D, et al. Heart Disease and Stroke Statistics 2010 Update: A Report From the American Heart Association Circulation 2010, 121:e46-e215. (2) Wong CK, et al. Hypocalcemic myocardial dysfunction: short- and long-term improvement with calcium replacement Am Heart J. 1990;120(2):381. (3) Braunwald[apos]s Heart Disease - A Textbook of Cardiovascular Medicine, 9th edition. (4) Tomar M, et al. Myocardial Dysfunction Due to Hypocalcemia Indian Pediatr. 2010 Sep;47(9):781-3. (5) Levine SN, Rheams CN. Hypocalcemic heart failure. Am J Med 1985; 78: 1033-1035.[br][br]Nothing to Disclose: NU, SRA, JSS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 840 187 1372 SUN-344 PO06-01 Sunday 1168 2012


1166 ENDO12L_SUN-345 POSTER SESSION: Parathyroid Hormone Disorders (1:30 PM-3:30 PM) A Unique Presentation of Normocalcemic Hyperparathyroidism with a Literature Review Alan N Peiris, Kamesh Sivagnanam, Rishika Motiani ETSU, Johnson City, TN; ETSU, Johnson City, TN Normocalcemic Hyperparathyroidism is an emerging entity and can present a diagnostic dilemma in the evaluation of patients suspected of having primary hyperparathyroidism. We report a patient that had intermittent elevation of ionized calcium levels along with a pronounced elevation of parathyroid hormone levels. We also discuss a clinical approach to this emerging entity and suggest a diagnostic and therapeutic algorithm.[br]A 66 year old Caucasian Veteran presented to us with elevated calcium and PTH levels. The patient had PTH levels ranging 300-900pg/ml [Normal 11-50pg/ml] and total calcium between 10.4-10.6mg/dl [normal 8.4-10.2mg/dl]. Serum albumin was between 3.9-4.3g/dl [Normal 3.5-5.0g/dl]. His ionized calcium level was 5.6mg/dl [Normal 4.5-5.6mg/dl], urinary calcium was 252mg/24 hours (normal 100-300mg/24 hours) and a 25(OH)D level of 13ng/ml [Normal 30-100ng/ml]. He received Vitamin D supplementation and in 2 years, total calcium levels improved to 8.6-10 mg/dl with normal ionized calcium but his PTH remained high. He was managed conservatively with repeated measurements of calcium and PTH and remained normocalcemic with elevated parathyroid hormone levels. This patient is of particular interest because his PTH levels were elevated to almost 6-25 times normal but still had normocalcemia and remained asymptomic.[br]Since multiple factors can influence calcium levels, a normal calcium level at least 60% of the times, with elevated PTH in the absence of secondary causes would be a good definition of NHP. The presence of normal ionized calcium levels augment the diagnosis and may represent [ldquo]true[rdquo] NHP. Measurement of ionized calcium is more sensitive than total calcium to identify NHP, but this can miss patients with NHP with normal ionized calcium. In this setting, PTH level may be the best screening tool.[br]Only a fraction of patients with NHP have been shown to develop PHP with hypercalcemia in prospective studies. However, NHP by itself is known to have musculoskeletal, cardiac and renal effects and is a condition that needs intervention. The management of patients with asymptomatic NHP with normal ionized calcium is however, best conservative. Follow up with labs consisting of intact PTH and ionized calcium is recommended. Also, there are no guidelines about when radiological testing is indicated and if it needs to be done more frequently or at earlier ages in this high risk group.[br][br]Nothing to Disclose: ANP, KS, RM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2340 187 1373 SUN-345 PO06-01 Sunday 1169 2012


1167 ENDO12L_SUN-346 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Rare Missense Variant in [italic]WDR5[/italic] in a Family with Novel Skeletal Dysplasia Characterized by Dysplastic Thumbs and Short Stature Andrew Dauber, Christina M Jacobsen, Joel N Hirschhorn Children[apos]s Hospital Boston, Boston, MA; Broad Institute, Cambridge, MA; Children[apos]s Hospital Boston, Boston, MA Background: Wdr5 has been shown to be necessary [italic]in vitro[/italic] for the differentiation of osteoblasts into mature osteoblasts (1). In a chick model, shRNA knock down of Wdr5 delays endochondral bone development resulting in impaired chondrocyte maturation and shortening of skeletal elements (2). There has been no human phenotype described thus far due to defects in [italic]WDR5[/italic] function. Whole exome sequencing allows for the rapid discovery of potentially causal genetic variants in individuals with rare genetic disorders.[br]Clinical Case: We describe a family with four individuals with dysplastic thumbs across three generations. The first affected female (adult height 149 cm, -2.5 SD) was born with no bones in one thumb and an inability to fully flex her second thumb. One of her two daughters also has limited flexion of both thumbs (adult height 151 cm, -2.1 SD). Two of her three sons are affected. Her first son (adult height 156 cm, -2.4 SD) has a hypoplastic 1[sup]st[/sup] metacarpal of his left thumb with an inability to fully flex it. The other son had a complete duplication of the phalanges in his right thumb (current height +0.2 SD).[br]Based on the pedigree, we hypothesized that the causal genetic variant is inherited in a dominant fashion and is likely quite rare and highly penetrant. Whole exome sequencing was performed in the three brothers. 3014 heterozgyous variants were shared by the 2 affected brothers and not by the 3[sup]rd[/sup] unaffected brother. We excluded all variants present in any prior database including dbSNP, 1000 Genomes, and NHLBI exome variant server leaving 29 candidate variants. We then tested these variants for segregation with phenotype in the unaffected aunt and affected grandmother. Nine variants segregated, three of which were nonsynonymous. One of the three variants was located in a gene, [italic]WDR5[/italic], with a known connection to skeletal biology. The missense variant (p.W114R) is in a highly conserved residue and is predicted to be damaging by Polyphen2.[br]Conclusion: We describe a family with a novel skeletal dysplasia involving dysplastic thumbs in four individuals, three of whom have significant short stature. All affected family members have a novel private missense variant in [italic]WDR5[/italic], a gene known to affect chondrocyte differentiation and osteoblastic maturation. Functional follow up studies are ongoing to further elucidate the effects of this variant [italic]in vitro[/italic].[br][br](1)Zhu ED, Demay MB, Gori F. Wdr5 is essential for osteoblast differentiation. J Biol Chem. 2008;283(12):7361[ndash]7. (2)Zhu S, Zhu ED, Provot S, Gori F. Wdr5 is required for chick skeletal development. J Bone Mineral Research 2010:25(11):2504-2514.[br][br]Sources of Research Support: NIH Grant 5K12HD052896-05 awarded to AND; Genentech Center for Clinical Research in Endocrinology awared to AND; March of Dimes Grant 6-FY09-507 awarded to JNH.[br][br]Nothing to Disclose: AD, CMJ, JNH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1008 188 1374 SUN-346 PO46-01 Sunday 1170 2012


1168 ENDO12L_SUN-347 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) A Novel [italic]GALNT3[/italic] Mutation Associated with an Unusually Severe Case of Familial Tumoral Calcinosis Jaydira Del Rivero, William H Chong, Diala El-Maouche, Felasfa Wodajo, Nisan Bhattacharyya, Rachel I Gafni, Alison M Boyce, Beth Brillante, Apurva N Trivedi, Runsheng Wang, Cailin Sibley, Marcus Y Chen, Alfredo Molinoto, Theo Heller, Raphaela Goldbach-Mansky, Adele Boskey, Kenneth E White, Michael T Collins National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; Weill Medical College of Cornell University, New York, NY; Indiana University School of Medicine, Indianapolis, IN; Virginia Hospital Center, Arlington, VA [bold]Introduction:[/bold] Homozygous inactivating mutations in [italic]GALNT3 [/italic]lead to increased processing of intact fibroblast growth factor 23 (FGF23) causing familial tumoral calcinosis (FTC), a rare disease characterized by decreased intact FGF23 (iFGF23), increased c-terminal FGF23 (cFGF23), hyperphosphatemia due to excessive tubular reabsorption of phosphate (TRP) and soft tissue calcifications. [bold]Case:[/bold] This 30 y/o Jordanian male first noted soft tissue calcifications at 10 y/o. Despite multiple debulking procedures, he had massive recurrences causing impaired mobility. Other affected family members reported much milder disease. Imaging showed extensive calcifications of the chest wall, extremities, colon, testes, thyroid and cerebrum but only trivial vascular calcifications. Labs showed phosphorus=5.5 mg/dl (2.5-4.8), calcium=8.5 mg/dl (8.2-10), 1,25 vitamin D3=74 pg/ml (18-64), PTH=55 pg/ml (16-87), cFGF23=572.5 RU/ml ([le]180) and iFGF23=8 pg/ml (10-50). Anemia [hemoglobin=6 g/dl (13.7-17.5), MCV=67.5 fL (79-92.2)] and elevated C-reactive protein (CRP=155 mg/L ([lt]3)) were noted. TRP was 92% (85-95) and Ca-Pi product was 63.43 ([lt]60). Histopathology showed soft tissue calcification and ossification with massive monocyte infiltration consistent with inflammation. Infrared imaging analysis of the material showed an elevated mineral/matrix ratio, suggesting it was not bone-like and not forming on a collagenous matrix. Therapy with sevelamer, acetazolamide and ketoconazole was unsuccessful. Additional therapies included parathyroid hormone, iron infusions, anakinra (an IL-1 inhibitor) and debulking surgeries. Genetic testing showed a novel frame shift mutation in exon 8 of [italic]GALNT3[/italic]. CRP decreased with anakinra, but no clinical changes were seen. Anakinra was stopped due to infection and CRP rebounded. iFGF23 and cFGF23 increased after iron transfusions, but no significant change in ratio was seen. [bold]Conclusions:[/bold] We report a case of severe FTC due to a novel mutation in [italic]GALNT3[/italic] with previously unreported findings and response to novel treatments. While the mechanism of soft tissue calcification in FTC is poorly understood, the microscopic findings, markedly elevated CRP, and response to anakinra suggest that inflammation may play a role. Given the phenotypic variability within this family, it is unclear whether this novel mutation caused his severe phenotype; however, [italic]GALNT3[/italic] is expressed in the GI tract and testes suggesting a role for [italic]GALNT3[/italic] in local inhibition of calcification.[br][br]Nothing to Disclose: JDR, WHC, DE-M, FW, NB, RIG, AMB, BB, ANT, RW, CS, MYC, AM, TH, RG-M , AB, KEW, MTC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1367 188 1375 SUN-347 PO46-01 Sunday 1171 2012


1169 ENDO12L_SUN-348 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Severe Infantile Hypophosphatasia Secondary to a Novel Compound Heterozygous Mutation Presenting with Pyridoxine-Responsive Seizures Dina Belachew, Traci Kazmerski, Arcangela Balest, Susan Stevens, Goldstein Amy, Jerry Vockley, Ingrid Libman Children[apos]s Hospital of Pittsburgh of UPMC, Pittsburgh, PA; Children[apos]s Hospital of Pittsburgh of UPMC, Pittsburgh, PA; Children[apos]s Hospital of Pittsburgh of UPMC, Pittsburgh, PA; Children[apos]s Hospital of Pittsburgh of UPMC, Pittsburgh, PA [bold]Background:[/bold][br]Hypophosphatasia (HPP) is a rare metabolic disease defined by reduced serum tissue non-specific alkaline phosphatase (TNSALP) activity. TNSALP is known for its role in mineralization. TNSALP is also active in the cerebral cortex. In severe HPP, defective TNSALP activity in the cerebral cortex has been associated with pyroxidine responsive seizures (PRS)[sup] 1[/sup]. This is hypothesized to be due to defective conversion of pyridoxal-5-phosphate (PLP) to pyridoxine. This step is necessary for the vitamin to enter cortical cells where it is a co-factor for numerous enzymes including regulators of GABA synthesis. An imbalance between excitation and inhibition then results in seizures[sup]2-5[/sup].[br][bold]Clinical Case:[/bold][br]A 35 week preterm Caucasian male infant presented with neonatal seizures at 18 hours of life. Clinical examination findings were normal. Due to intrauterine exposure to opiates and maternal HSV, neonatal abstinence syndrome and neonatal HSV were initially suspected and treated. However, seizures persisted despite conventional anti-epileptic medications. At two months of age, empiric treatment with favorable response to pyridoxine unfolded the diagnosis of PRS. Additionally, diffuse bone mineralization defects, hypercalcemia, hypercalciuria, nephrocalcinosis and undetectableserum alkaline phosphatase led to suspicion of HPP. Laboratory data showed elevated urine elevated phosphoethanolamine (PEA) and significantly elevated CSF PLP. Definitive diagnosis was made by sequence analysis of the TNSALP gene, which revealed compound heterozygosity with novel mutation in exon 9, c.875_881delCAGGGGAinsT involving a large deletion, and a previously reported mutation in exon 12, c.1559delT. These mutations confer severe disease ultimately observed in our patient. However, a delay in the development of overt phenotypic features of HPP implies that phenotype-genotype correlation could be variable even in severe disease.[br][bold]Conclusions:[/bold][br]This case illustrates that careful scrutiny of laboratory data and continual revision of differential diagnosis is crucial in reaching the diagnosis of rare entities like HPP. HPP must be considered in the differential of PRS and ALP must be measured in these patients. Therapy requires a multidisciplinary approach addressing feeding disorders, respiratory compromise, craniosynostosis and increased intracranial pressure, hypercalcemia and developmental delay. Close follow-up is critical.[br][br](1)Baumgartner-Sigl et al.,Bone 2007;40(6):1655-61. (2)Plecko et al., Can J Neurol Sci 2009;36 Suppl 2:S73-7. (3)Stockler et al.,Mol Genet Metab 2011; 104 (1-2): 48-60. (4)Nunes et al., J Child Neurol 2002;17(3):222-4. (5)Balasubramaniam et al., J Inherit Metab Dis 2010.[br][br]Nothing to Disclose: DB, TK, AB, SS, GA, JV, IL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2176 188 1376 SUN-348 PO46-01 Sunday 1172 2012


1170 ENDO12L_SUN-349 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Albright Hereditary Osteodystrophy (AHO) and Progressive Osseous Heteroplasia (POH): Overlapping Features and Clinical Presentation Maria H Lin, Pisit Pitukcheewanont Children[apos]s Hospital Los Angeles, Los Angeles, CA Introduction: POH is a condition of heterotopic ossification. Case reports of patients with mild POH and features of AHO, specifically pseudohypoparathyroidism (PHP) type 1a with hormonal resistance, suggest the possibility of a common molecular basis. The [italic]GNAS[/italic] gene has been implicated to account for the overlapping phenotypic spectrum of POH and AHO (specifically PHP type 1a).[br][italic]Case 1[/italic]: 4-year-old boy presented with obesity, speech delay, and expanding subcutaneous masses on his buttock/forearm. Physical exam revealed round facies, brachydactyly, and erythematous plaques over right upper thigh/forearm. Blood tests showed normal Ca, P, Mg, 25(OH)D levels but elevated intact PTH (369 pg/mL, n[lt]65 pg/mL), ALP (285 U/L, n[lt]220 U/L), and TSH (8.3 mIU/mL, n[lt]5). Bone age was advanced by 1 year. Abdominal CT showed multiple calcifications in subcutaneous tissue, fat, and muscle. Pathology of excised tissue revealed ossification. Genomic study revealed no [italic]GNAS[/italic] gene mutation. He was diagnosed with POH with AHO features and treated with calcitriol and levothyroxine.[br][italic]Case 2[/italic]: 3-year-old boy presented with extensive, painful ossification in his left lower extremity. No round facies or brachydactyly was noted. He had normal Ca, P, Mg, and 25(OH)D levels but elevated intact PTH 107 pg/mL, ALP 324 U/L, and high-normal TSH 4.04 mIU/mL. Bone age was advanced by 1 year. CT scan showed subcutaneous and intramuscular calcifications in left lower extremity. Genetic testing showed a mutation in exon 12 of the [italic]GNAS[/italic] gene. Patient had features of POH and PHP type 1a and was treated with calcitriol.[br][italic]Case 3[/italic]: 9-year-old boy presented with knee pain and diffuse subcutaneous ossification in his back and upper/lower extremity. He had significantly limited joint mobility and was non-ambulatory. He had no round facies or brachydactyly. Lab tests showed normal Ca, P, Mg, 25(OH)D, intact PTH, and TSH levels. Bone age was not advanced. No [italic]GNAS[/italic] mutation was found. CT scan showed subcutaneous and intramuscular calcifications, distributed throughout his shoulders, torso and extremities. He was diagnosed with POH.[br]Clinical Lessons: Patients with heterotopic ossification present with a wide spectrum of disease. Although [italic]GNAS[/italic]-based mutations have been postulated to account for the overlapping features of AHO and POH, normal DNA studies in certain patients with POH/AHO suggest that there may exist other molecular/epigenetic mechanisms that explain the overlapping phenotypic features for these conditions.[br][br]Nothing to Disclose: MHL, PP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 540 188 1377 SUN-349 PO46-01 Sunday 1173 2012


1171 ENDO12L_SUN-350 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Identification and Functional Characterization of a Novel [italic]NPR2[/italic] Mutation (p.R388Q) Causing Acromesomelic Dysplasia, Type Maroteaux Naoko Amano, Hiroshi Kito, Satoshi Narumi, Rumi Hachiya, Yoshihiro Ogawa, Gen Nishimura, Tomonobu Hasegawa Keio University School of Medicine, Tokyo, Japan; Keio University, Tokyo, Japan; Nagoya University School of Medicine, Aichi, Japan; Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan; Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan; Tokyo Metropolitan Children[apos]s Medical Center, Tokyo, Japan [bold]Background: [/bold]Acromesomelic dysplasia, type Maroteaux (AMDM) is a rare congenital bone dysplasia, which is characterized by disproportionate shortening of skeletal elements, predominantly affecting the middle and distal segments of the appendicular skeleton with an autosomal recessive inheritance. AMDM is caused by loss-of-function mutation of [italic]NPR2[/italic]. NPR-B that is coded by [italic]NPR2[/italic], is the guanylyl cyclase-coupled receptor of C-type natriuretic peptide (CNP). CNP-NPR-B pathway has an important role to stimulate endochondral ossification, which leads to long bone growth. [bold]The Patient: [/bold]The proband was a 25-year-old Japanese female, who was born at term as a child of healthy parents, who are first cousins. Birth length and weight was 46.6cm (10th percentile) and 3,570g (90-97[sup]th[/sup] percentile), respectively. At 2 years of age, she was noted to have short stature. At 21 years of age, her height and weight was 119.0 cm (-7.8 SD), and 35 kg (-2.3 SD). Physical examination showed acromesomelic shortening and brachydactyly. On radiological examination, her metacarpals and phalanges were short and wide, and the heights of her vertebrae were short. We diagnosed her as having AMDM.[bold] Methods and Results:[/bold] We analyzed all coding exons and flanking introns of [italic]NPR2 [/italic]by PCR-based sequencing. We identified a novel homozygous missesnse mutation (c.1163G[gt]A, p.R388Q) in the proband. Her parents had the mutation in a heterozygous state. The mutation was not detected in 100 control individuals. Arg388 is highly conserved among vertebrate species and in human NPR-A. To evaluate guanylyl cyclase activity, we examined cGMP production by CNP treatment in the cells transfected with either HA-wild (wild type) or HA-R388Q (mutant) NPR-B. The cGMP concentration in the cells transfected with wild type was 21.4 [plusmn] 1.51 pmol/well. However there was no statistically significant difference between mock (0.46 [plusmn] 0.11 pmol/well) and the mutant (0.34 [plusmn] 0.05 pmol/well). By Western blot, the expression of the mutant was equivalent to that of the wild type. To assess intracellular localization of the mutant, we performed immunofluorescent studies using an anti-HA antibody. While wild type exclusively displayed cell surface expression, the mutant was entrapped in endoplasmic reticulum and was poorly expressed in the cell surface. [bold]Conclusions: [/bold]We identified a novel [italic]NPR2[/italic] mutation (c.1163G[gt]A, p.R388Q) in a patient with AMDM. The functional pathogenesis of this mutation is defective trafficking to the cell surface.[br][br]Nothing to Disclose: NA, HK, SN, RH, YO, GN, TH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1213 188 1378 SUN-350 PO46-01 Sunday 1174 2012


1172 ENDO12L_SUN-351 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Full Description of Bone Metabolism in a Case of Morquio-Brailsford Syndrome Catherine Adam, Agnes Rakel, Louis-George Ste-Marie Centre de Recherche du CHUM, Universit[eacute] de Montr[eacute]al, Montr[eacute]al, Canada Morquio-Brailsford[apos]s syndrome is a rare autosomal recessive lysosomal storage disorder in which keratan sulphate (KS) metabolism is impaired, resulting in an accumulation of KS in specific tissues, alteration of bone structure and characteristic skeletal abnormalities. The case involves a patient with Morquio[apos]s syndrome who was first seen in our clinic at age 37, in 1999. She had had bilateral hip replacement at age 17 but never suffered any fracture. The blood work showed normal levels of calcium, phosphorus and albumin. The C-terminal PTH level was on the upper limit of normal. The serum bone-specific alkaline phosphatase, as well as the 24-hour urinary calcium were normal. The 24-hour urinary C-telopeptides were slightly increased. Plain x-rays showed classic findings of Morquio[apos]s syndrome such as lumbar scoliosis, general platyspondyly, and degenerative changes in the shoulders and in the 2nd and 3rd metacarpal epiphysis. Quantitative uptake during a whole body bone scan showed an increased CR-51/Tc-99m ratios consistent with mild bone hypermetabolism. A DEXA of the lumbar spine showed a bone mineral density (BMD) of 0.715g/cm2 at L1-L4 (T score = -4.0). A transiliac bone biopsy showed normal trabecular architecture with very low trabecular volume (7.8%, NI=23.3[plusmn]4.1%), normal eroded surfaces (3.2%, NI=3.6[plusmn]1.1%), normal mineralization rate (0.85[micro]m/d, NI=0.72[plusmn]0.12 [micro]m/d) and slightly increased remodelling. Following those results, the patient was put on calcium, vitamin D and oral bisphosphonates (BP). A follow-up evaluation 10 years later at age 50 revealed an improvement of the lumbar spine BMD [L1-L4 = 0.772g/cm2 (T score = -3.4)]. The patient reported no history of fractures during the period of time between the start of the treatment and the evaluation. To our knowledge, this is the first complete description of bone metabolism of Morquio[apos]s syndrome including remodelling marker, bone mineral density, quantitative uptake during a whole body bone scan and bone biopsy. Despite a very low BMD, the patient had no history of fractures while treated during ten years with an oral bisphophonate.[br][br]Nothing to Disclose: CA, AR, L-GS-M 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1805 188 1379 SUN-351 PO46-01 Sunday 1175 2012


1173 ENDO12L_SUN-352 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Lowe Syndrome, Related Fanconi Syndrome, Hypogonadism and Resultant Multi-Factorial Bone Disease Randall Fenner, Richard Comi Dartmouth-Hitchcock Medical Center, Lebanon, NH Background: Lowe[apos]s syndrome (oculocerebrorenal syndrome of Lowe, LS) is a rare X-linked disorder characterized by bilateral cataracts and hypotonia at birth, Fanconi syndrome within the first few months, motor and mental developmental delays, and facial dysmorphisms. Fanconi syndrome can cause rickets, osteomalacia, and fractures if left untreated and this is further complicated if other risk factors are present.[br]Clinical Case: A 19 (now 20) year man presented with his second seizure related femoral fracture within a year. LS was diagnosed early in life. Cataracts were removed shortly after birth and he had delayed mental (currently cognitively impaired) and motor development (not walking until age 4). He is ambulatory for short distances since the fractures. Associated Fanconi syndrome was well treated since diagnosis, including calcitriol, citric acid- sodium citrate and potassium phosphate. Seizures (also common in LS) started at age 2 and were treated with carbamazepine or phenobarbital for 15 years. Currently he is on Keppra and Lamictal. Cryptorchidism (also mentioned in LS literature) was diagnosed at young age and he underwent unilateral orchiectomy and contralateral orchiopexy. Subsequently he did not undergo puberty. Bone age performed at chronologic age 16.5 years was 15.5 years and showed significant osteopenia. Other x-rays showed open growth plates. DXA scan showed total lumbar spine T score of -5. Initial lab tests showed: CO2 21 mMol/L (22-31), Cr 0.81 mg/dL (0.8-1.5), Ca 8.9 mg/dL (8.5-10.5), PTH 18 pg/ml (15-65), 25-Vit D 29 ng/ml (25-80), IGF-1 183 ng/ml (281-510), FSH 86.7 mIU/ml (1.5-12.4), LH 54.8 mIU/ml (1.7-8.6), total testosterone 0.37 ng/ml (2.8-8), Mg 0.95 mMol/L (0.69-1.07), and Phos 2.5 mg/dL. Options included hormonal (testosterone or estrogen) and standard osteoporosis medications. After discussion with parents it was decided to treat with slow titration of testosterone gel (no injections or peaks, and small incremental increases possible) therapy at 1.25 gm every other day and current dose is alternating days of 1.25 and 2.5 gm. Recent total testosterone is 164 ng/dL (250-1100) and signs of pubertal progression are present. He has not had any worsening of behavioral issues nor fractures since starting testosterone therapy.[br]Conclusion: Multi-factorial bone disease in Lowe[apos]s syndrome is complex and the first step may be to correct current deficiencies or biochemical abnormalities before moving to further intervention.[br][br]Nothing to Disclose: RF, RC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1604 188 1380 SUN-352 PO46-01 Sunday 1176 2012


1174 ENDO12L_SUN-353 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Calcium-Sensing Receptor Activation or Hypoparathyroidism? Naveen Hullekere Siddaramaiah, Margaret Morris, Timothy Cheetham, Simon Pearce Royal Victoria Infirmary, Newcastle-upon-Tyne, UK Gain-of-function mutations in the calcium-sensing receptor (CASR) are associated with a familial syndrome of hypocalcaemia with hypercalciuria. Over 20 activating mutations have been identified, causing familial autosomal dominant hypocalcaemia[sup]1-6[/sup]. Therapy with activated vitamin D analogues can cause hypercalciuria and nephrocalcinosis, with poor correlation between serum [amp] urine calcium levels[sup]7[/sup].[br]We present one family and a sporadic case with biochemical features suggestive of Calcium Sensing receptor activation, but negative for the mutations.[br][bold]Family [/bold][br][bold]Proband: 53yr old man[/bold], presented at 30yrs with asymptomatic hypocalcaemia (Ca[sub]adj[/sub]1.63mmol/l, Ref 2.10-2.60) and hypomagnesaemia (0.62mmol/l), with low PTH (11ng/l, Ref 15-65) and a urine calcium/creatinine ratio of 0.09mmol/mmol(0- 0.7). He became symptomatic during calcitriol therapy, which was associated with hypercalciuria (ranging 0.22 to 0.94 mmol/mmol). Trial reduction in calcitriol caused severe muscle spasms, not keeping with calcium receptor problems. Serum PTH level remained low to normal ([lt]2-14ng/L). His daughter presented at 16yrs, with minimal symptoms of hypocalcaemia (Ca[sub]adj[/sub]-1.49mmol/l) and low normal PTH (8ng/L). Urine calcium/creatinine ratio was 0.27mmol/mmol. DNA sequencing of the calcium-sensing receptor (exons 2 to 7) and cytogenetic analysis including TUPLE FISH probe for 22q11.1 was normal. His 25yr old son also has asymptomatic hypocalcaemia.[br][bold]Case 2:[/bold][br]Female (26yrs). She presented with carpopedal spasm at the age of 3yrs with a Ca[sub]adj [/sub]of 1.14mmol/L, Mg[sup]2+ [/sup]of 0.62mmol/L, and PTH [lt]10ng/L (10-65). Her parents are normocalcaemic. She was treated with calcidiol (alfacalcidol) and had normal growth and development. She remained asymptomatic despite withdrawal of calcidiol for ten years from the age of 11. She then presented with tiredness, thirst and polyuria. Adjusted calcium level have remained in the range of 1.36 to 2.25mmol/L, on replacement, with hypercalciuria (0.74 to 1.32mmol/L) and low PTH levels ([lt]2 to 6ng/L). DNA sequencing of the calcium-sensing receptor (exons 2 to 7) and cytogenetic analysis was normal.[br][bold]Conclusion:[/bold] It may be difficult to distinguish patients with calcium-sensing receptor activating mutation from those with true idiopathic hypoparathyroidism. Available genetic analysis has not identified any mutation in these cases, despite significant clinical pointers to the former diagnosis. The cause of hypocalcaemia in these patients remains uncertain.[br][br]1. A Familial syndrome of hypocalaemia with hypercalciuria due to mutations in the calcium-sensing receptor. Simon H.S.Pearce et al. NEJM 1996;335(15):1115-22. 2. Activating mutations of the calcium-sensing receprtor: management of hypocalcaemia. Anne Lienhardt et al. JCEM 2001;86(11):5313-23. 3. A family of autosomal dominant hypocalcaemia with a positive correlation between serum calcium and magnesium: Identification of a novel gain of function mutation (Ser820Phe) in the calcium-sening receptor. Terumasa Nagase et al. JCEM 2002;87(6):2681-87. 4. Dominant-negative GCMB mutations cause an autosomal dominant form of hypoparathyroidism. Michael Mannstadt et al. JCEM 2008; 93(9):3568-76. 5. Familial hypoparathyroidism: Identification of a novel gain of function mutation in transmembrane domain 5 of the calcium-sensing receptor. Tomoyuki Watanabe et al. JCEM 1998;83(7):2497-502. 6. A novel activating mutation in calcium-sensing receptor gene associated with a family of autosomal dominant hypocalcaemia. Ryo Okazaki et al. JCEM 1999;84(1):363-66. 7. Mutations in the calcium-sensing receptor gene cause autosomal dominant and sporadic hypoparathyroidism. Jeffrey Baron et al. Hum Mol Gene 1996;5(5):601-6.[br][br]Nothing to Disclose: NHS, MM, TC, SP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2287 188 1381 SUN-353 PO46-01 Sunday 1177 2012


1175 ENDO12L_SUN-354 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Primary Hyperparathyroidism and Familial Hypocalciuric Hypercalcemia, a Diagnosis Made 15 Years after Parathyroidectomy Noureddine Moumli, Pierre MJ Zelissen, B Bravenboer UMC, Utrecht, Netherlands; Catharina Ziekenhuis, Eindhoven, Netherlands Case: A 54-year-old man was referred for persistent hypercalcaemia since 13 years despite extensive neck surgery. He had no specific symptoms. Neck ultrasound and sestamibi-scintigraphy showed no abnormalities.[br]In 1998 patient was diagnosed with PHPT. Imaging of the neck was inconclusive. Repeated surgical exploration was performed, resecting the thymus and 3 parathyroids, all histologically hyperplastic. Postoperatively there was in all occasions transient normocalcaemia, with recurrence of the hypercalcaemia within weeks. The following years different tests were performed: DNA-sequencing for MEN-1 (negative) because of the 3 parathyroid [ldquo]adenoma[apos]s[rdquo], 2 MIBG scans because of palpitations (no feochromocytoma), DEXA-scanning (no osteoporosis) and abdominal CT (no nephrolithiasis).[br]Consecutive laboratory tests showed a mildly elevated calcium (2.70-2.75 mmol/L, N 2.20-2.60), low phosphorus (0.73 mmol/L, N 0.80-1.50), an inappropriately normal PTH (4-8 pmol/L, N 1.0-7), normal vit.D level; normal to slightly elevated 24h urinary calciuria (4.0-7.8 mmol/24h, N 2.50-7.50) and a calcium/creatinine clearance ratio of 0.008-0.012.[br]Due to the lack of symptoms, absence of manifestations of longstanding PHPT, and recurrence of hypercalcaemia we doubted the initial diagnosis and considered FHH. Sequencing of the CaSR gene showed a novel heterozygous mutation in exon 6 (c.1588T[gt]C). Patient had no available relatives to confirm the pathogenicity of this mutation. We performed a literature search and found another patient with a mutation in exact the same base, but in this case from T to G (c.1588T[gt]G). In vitro molecular testing confirmed impaired CaSR-function by expressing mutant and wild-type CaSRs in embryonic kidney cells and studying receptor-signalling in response to different concentrations of extracellular calcium.[br]Discussion: Differentiating PHPT from FHH can be challenging, because of the similarities in the biochemical and clinical presentation. The biochemical pattern in this case suggests PHPT, especially the absence of hypocalciuria. Nonetheless FHH should be considered. The term [ldquo]hypocalciuric[rdquo] in FHH is somewhat misleading. In recent literature up to 20% of patients with confirmed FHH had normal renal calcium excretion.[br]Patients with FHH don not benefit from surgical intervention. Establishing the right diagnosis can avoid unnecessary neck surgery and expensive tests.[br][br]Nothing to Disclose: NM, PMJZ, BB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2316 188 1382 SUN-354 PO46-01 Sunday 1178 2012


1176 ENDO12L_SUN-355 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Calcium-Sensing Receptor (CaSR): A Key Player in Calcium Homeostasis Deepashree Gupta, Camila Margharita Manrique University of Missouri-Columbia, Columbia, MO [bold]Background[/bold]: To demonstrate importance of Calcium Sensing Receptor (CaSR) in calcium homeostasis by contrasting activating and inactivating mutations of CaSR[br][bold]Clinical Case[/bold]: 63 year old Caucasian female who presented to Endocrinology clinic with bloating, constipation, irritability and [apos]face feeling like a mask[apos]. She was diagnosed with hypoparathyroidism at age of 22 and started on calcium and calcitriol replacement. She underwent mutation analysis at Johns Hopkins Hospital but was lost to follow up. At presentation, she was taking calcium citrate 500 mg TID and calcitriol 0.25 mcg BID. She had history of nephrolithiasis which resolved with hydrochlorothiazide use. Two of her children, her brother, his two children and her mother also have hypoparathyroidism. Physical exam did not reveal signs of hypocalcemia. Labs showed 25 OH vitamin D level-83 ng/ml (30-80), iPTH-7 pg/ml (15-65), calcium-9.1 mg/dl (8.5-10.5), albumin-4 g/dl (3.4-5), magnesium-1.8 mg/dl (1.6-2.6), phosphorus-3.7 mg/dl (2.7-4.5). Mutation analysis was positive for calcium sensing receptor activating mutation. Her current therapeutic regimen is calcium citrate 250 mg TID and calcitriol to 0.5 mcg once daily. Most recent calcium level is 7.9 mg/dl and she is symptom free.[br]22 year old Caucasian male was first seen by pediatric endocrinology at age of 14 for hypercalcemia (Ca-11.7 mg/dl). Since he had no symptoms and only mildly elevated ionized calcium, normal PTH and hypocalciuria, it was decided to observe him. His new PCP referred him to adult endocrinology clinic to continue follow up for hypercalcemia. There is a strong family history of hypercalcemia in paternal grandmother and paternal aunts. Patient was not using any offending medications. Physical exam was normal. Calcium-12 mg/dl (8.5-10.5), phosphorus-2.3 mg/dl (2.7-4.5), intact PTH-38.5 pg/ml (15-65), 24-hour urine calcium-86 mg/24 hour, calcium to creatinine clearance ratio-0.002 and 25 hydroxy vitamin D level-29 ng/ml. Given that he had hypercalcemia since childhood which remained stable, low normal PTH, low urinary calcium and family history of hypercalcemia, Familial Hypocalciuric Hypercalcemia (FHH) was diagnosed; reassurance was provided with regular follow up of calcium levels.[br][bold]Conclusion: [/bold]Familial hypoparathyroidism and FHH are caused by activating and inactivating mutations of the CaSR respectively. These cases illustrate the different mutations in CaSR that result in abnormalities in calcium homeostasis.[br][br]Nothing to Disclose: DG, CMM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1259 188 1383 SUN-355 PO46-01 Sunday 1179 2012


1177 ENDO12L_SUN-356 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) A Novel Calcium-Sensing Receptor Gene ([italic]CASR[/italic]) Variant Associated with Hypocalciuric Hypercalcemia Anu Vishwanath, Julie Beasley, Sowmya Krishnan, Klaas Wierenga University of Oklahoma Health Sciences Center, Oklahoma City, OK; University of Oklahoma Health Sciences Center, Oklahoma City, OK [bold]Background [/bold][br]The calcium sensing receptor (CaSR) regulates parathyroid hormone (PTH) secretion by the parathyroid glands and calcium regulation in the kidney and other tissues. Both activating and inactivating mutations in [italic]CASR[/italic] have been described leading to variable clinical presentation ranging from hypocalcemia to hypercalcemia.[br][bold]Clinical Case [/bold][br]A 16.5 year old Ukrainian male was referred for evaluation of hypercalcemia noted on routine labs with Calcium (Ca) of 11.5 mg/dL (8.4-10.2) and ionized Ca 1.46 mmol/L (1.12-1.32). He did not have symptoms typical of hypercalcemia and was on medications for behavioral problems. Limited family history was available as he was adopted. Further diagnostic work-up noted serum phosphorous (2.8 mg/dL (2.5-4.5)), PTH (34 pg/mL (7-53)), 1, 25-dihydroxyvitamin D (69 pg/mL (15 [ndash] 90)), 25-hydroxyvitamin D (26 ng/mL (32 [ndash] 100)) and a slightly elevated magnesium (2.3 mg/dl (1.7-2.2)). Urinary studies revealed a fractional excretion of Ca (FE[sub]Ca[/sub]) of 0.007 ([gt]0.01), highly suggestive for hypocalciuria. Imaging studies noted a normal renal ultrasound and a DXA scan revealed normal bone mineral density both in total body (1.103 g/cm2, z-score of -0.4) and AP spine (1.131 g/cm2, z-score of -0.7). The patient[apos]s clinical status and labs were consistent with familial hypocalciuric hypercalcemia (FHH). [italic]CASR[/italic] gene sequencing revealed a heterozygous c.1373 G[gt]T transversion in exon 4, predicted to result in p.Trp458Leu substitution. This novel missense mutation is located in the extracellular domain of CaSR, known to be involved in calcium sensing. The p.Trp458 is highly conserved across species from human to zebrafish, and mutations at this region are predicted to be damaging by bioinformatic analyses. Review of literature shows that most mutations in [italic]CASR[/italic] associated with FHH are point mutations clustered either in the extracellular calcium-sensing domain, encoded particularly by exons 3 and 4, or in the signal-transducing domain, encoded by exon 7.[br][bold]Implications for practice[/bold][br]It is important to differentiate hypercalcemia due to FHH from primary hyperparathyroidism (PHPT) as the two diagnoses entail different management strategies with parathyroidectomy recommended in PHPT while FHH is usually a benign condition. Further functional studies to characterize the effect of this substitution and identifying other FHH patients who may harbor this mutation, may provide corroborating evidence to establish a causative role in FHH.[br][br]Nothing to Disclose: AV, JB, SK, KW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1861 188 1384 SUN-356 PO46-01 Sunday 1180 2012


1178 ENDO12L_SUN-357 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Familial Hypocalcemia [mdash] Think about Calcium Sensor Receptor (CaSR) David Finn, Marianna Stephanie How Yaw, Verghese Mathew, Katrina Rachel Prescott, Sanjay Gupta Hull Royal Infirmary, Hull, UK; Chapel Allerton Hospital, Leeds, UK [bold]Aim: [/bold]Activating mutations of CaSR gene have been shown to cause hypercalciuric hypocalcaemia[bold][sup]1[/sup][/bold]. The CaSR plays a key role in maintaining calcium homeostasis by regulating parathyroid hormone (PTH) and renal calcium reabsorption. Activating mutation of CaSR causes increased sensitivity to Ca++ and hence down regulation of PTH secretion.[br][bold]Method:[/bold] This is a case series where we report a pedigree with clinical picture consistent with familial autosomal dominant hypoparathyroidism. The proband, an 18-month-old girl was found to have incidental hypocalcaemia. We identified 4 other affected individuals in the family, including the father of the proband and his twin brother. All affected individuals had mild hypocalcaemia, low normal serum parathyroid levels and borderline high phosphate levels. Treatment with vitamin D and oral calcium did not improve their serum calcium levels. The father of proband has shown evidence of hypercalciuria and has recently been treated for urolithiasis. Mutation analysis identified a novel activating mutation of the CaSR gene [c.407C[gt]T(p.Pro136Leu)] in all 3 family members who were tested.[br][bold]Conclucion:[/bold] Activating mutation of the CaSR inhibits PTH secretion and renal calcium reabsorption despite hypocalcaemia. Treatment with vitamin D and calcium can increase hypercalciuria leading to nephrocalcinosis and renal impairment[bold][sup]1,2[/sup][/bold]. Asymptomatic individuals should not be overtreated in an attempt to normalise serum calcium.[br][br](1) Lienhardt A et al (2001) Activating mutation of the calcium-sensing receptor: management of the hypocalcaemia The Journal of Clinical Endocrinology and Metabolism 86(11):5313[ndash]5323. (2) Pearce SHS et al (1996) A familial syndrome of hypocalcemia with hypercalciuria due to mutations in the calcium-sensing receptor. The New England Journal of Medicine 335(15) 1115-1122.[br][br]Nothing to Disclose: DF, MSHY, VM, KRP, SG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2005 188 1385 SUN-357 PO46-01 Sunday 1181 2012


1179 ENDO12L_SUN-358 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) A Novel Mutation in Familial Hypocalciuric Hypercalcemia Presenting in Early Infancy with Irritability and Mild Hypercalcemia Nursen Gurtunca, Ediz Yesilkaya, Selma Witchel Children[apos]s Hospital of Pittsburgh of UPMC, Pittsburgh, PA; Gulhane Military Medical Academy, Ankara, Turkey The Calcium-sensing receptor (CaSR) belongs to the G-protein-coupled receptor superfamily and is expressed primarily in the parathyroid glands, C cells of the thyroid and the kidney. CaSR functions like a thermostat to help maintain ionized calcium level in the narrow physiologic range. Heterozygous inactivating mutations of the CaSR cause Benign Familial Hypocalciuric Hypercalcemia (BFHH). More than 200 inactivating mutations of the CaSR gene have been described to date.[br]BFHH is a rare genetic condition resulting in reduced ability of the CaSR to sense or respond to elevated calcium concentrations. Inactivation of the receptor (OFF-switch of the thermostat) causes excessive tubular resorption of calcium, resulting in hypercalcemia, relative hypocalciuria, and inappropriately high serum parathyroid hormone concentration. We are describing a novel mutation in CaSR gene in an eight week old female infant of immigrant parents of Turkish descent from Russia.[br]This 8 week old female infant presented with history of increasing irritability and poor feeding and assumed to have infantile colic. Investigations revealed hypercalcemia, hypocalciuria and inappropriately elevated parathyroid hormone. Her mother was screened for hypercalcemia and also found to have mild hypercalcemia, hypocalciuria and elevated parathyroid hormone. Screening of the father, two older sibling and genetic studies of both of the parents were carried out. A novel heterozygote insertion was detected in the patient and the mother. Insertion of G between base pair 108 and 109 in exon 2 results in a change of leucine to alanine at codon 37 and a frameshift creating a STOP codon at position 47 in one of the two CaSR alleles. Based on a previous report of a different heterozygous insertion that also results in a stop codon at position 47 in a family affected with BFHH (D[apos]Souza-Li, Yang et al. 2002), this mutation is likely to cause inactivation of the CaSR.[br]CONCLUSION: Most infants with BFHH are asymptomatic and the condition is usually diagnosed on routine testing later in life. In affected infants during periods of decreased fluid intake hypercalcemia may become more pronounced and lead to symptoms that may mimic infantile colic.[br][br]1. Nakayama T, Minato M, Nakagawa M, Soma M, Tobe H, Aoi N, Kosuge K, Sato M, Ozawa Y, Kanmatsuse K, Kokubun S. A novel mutation in Ca2+-sensing receptor gene in familial hypocalciuric hypercalcemia. Endocrine. 2001 Aug;15(3):277-82. 2. Egbuna OI, Brown EM. Hypercalcaemic and hypocalcaemic conditions due to calcium-sensing receptor mutations. Best Pract Res Clin Rheumatol. 2008 Mar;22(1):129-48. 3. Ward BK, Magno AL, Davis EA, Hanyaloglu AC, Stuckey BG, Burrows M, Eidne KA,Charles AK, Ratajczak T. Functional deletion of the calcium-sensing receptor in a case of neonatal severe hyperparathyroidism. J Clin Endocrinol Metab. 2004 Aug;89(8):3721-30. 4. Egbuna OI, Brown EM. Hypercalcaemic and hypocalcaemic conditions due to calcium-sensing receptor mutations. Best Pract Res Clin Rheumatol. 2008 Mar;22(1):129-48. 5. D[apos]Souza-Li L, Yang B, Canaff L, Bai M, Hanley DA, Bastepe M, Salisbury SR,Brown EM, Cole DE, Hendy GN. Identification and functional characterization of novel calcium-sensing receptor mutations in familial hypocalciuric hypercalcemia and autosomal dominant hypocalcemia. J Clin Endocrinol Metab. 2002 Mar;87(3):1309-18.[br][br]Nothing to Disclose: NG, EY, SW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 834 188 1386 SUN-358 PO46-01 Sunday 1182 2012


1180 ENDO12L_SUN-359 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) An Unusual Case of Sporadic Pseudohypoparathyroidism Ib with Osteitis Fibrosa Cystica Luisella Cianferotti, Agnese Biagini, Eduardo Fernandez-Rebollo, Harald Jueppner, Claudio Marcocci University of Pisa, Pisa, Italy; Massachusetts General Hospital - Harvard Medical School, Boston, MA; Massachusetts General Hospital - Harvard Medical School, Boston, MA Background: The term pseudohypoparathyroidism (PHP) refers to a heterogeneous group of disorders with resistance to PTH and frequently to other hormones activating cAMP-dependent responses downstream of various G protein-coupled receptors. PHP-Ia is caused by inactivating mutations involving GNAS exons encoding the G[alpha]s, while PHP-Ib is caused by deletions within or upstream of this locus.[br]Clinical case: We describe a 36-yr-old man affected by sporadic PHP-Ib with renal PTH-resistance, along with severe hyperparathyroid bone disease. The patient first presented at age 8 yrs with limping due to bone deformities; he had undergone multiple osteotomies. Baseline biochemical showed hypocalcemia (7.5 mg/dl) with elevated PTH (504 pg/ml) and increased ALP (487 IU/L). Upon restoring blood calcium level to the low-normal range by oral calcium (1 g/day) and calcitriol (0.5-1 [mu]g/day), an increase in PTH and ALP was observed (2100 pg/ml and 980 IU/L, respectively). Serum calcitonin was slightly elevated (22 pg/ml; normal: [lt]10) and was hyperresponsive to pentagastrin stimulation (225 pg/ml after 5 minutes). Cortical bone mineral density (BMD) was markedly reduced (distal radius T-score: -7.7). Subperiosteal bone resorption with acroosteolysis, salt-and-pepper skull, bone cysts and pseudotumors and long bone deformities were evident on radiographic studies. Parathyroid ultrasound was consistent with four-gland hyperplasia. After increasing calcitriol to 4 [mu]g/day, PTH and ALP decreased continously over the course of 8 months to 130 pg/ml and 97 IU/L, respectively, while serum and urinary calcium levels remained normal. Cortical BMD markedly increased within 6 months (+6.5%) and bone pain resolved. After mild, transient hypercalcemia developed, likely because of completion of the remineralization process, the calcitriol dose was reduced to 1.5 [mu]g/day and normocalcemia was attained.[br]Basic analyses: Analyses of the GNAS locus showed methylation changes at all four DMRs without evidence for paternal uniparental isodisomy of 20q (patUPD20q); the 3-kb STX16 deletion was excluded. No features of PHP-Ib were present in 1st degree relatives.[br]Conclusion: A PHP-Ib patient with long-standing hypocalcemia due to renal PTH-resistance associated with abnormal GNAS methylation showed marked bone resorption. After increasing oral calcitriol a dramatic decline in PTH levels close to the normal range was observed, which resulted in marked bone remineralization and much improved symptoms.[br][br]Sources of Research Support: NIH, NIDDK Grant R37DK46718 awarded to HJ.[br][br]Nothing to Disclose: LC, AB, EF-R, HJ, CM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2019 188 1387 SUN-359 PO46-01 Sunday 1183 2012


1181 ENDO12L_SUN-360 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) 4 Gland Hyperplasia Primary Hyperparathyroidism Picked up on Imaging in MEN 1 Case Aswathiah Srinath, Prakash Sinha, Adrian Thomas, Abbi Lulsegged Kings College Hospital, London, UK; South London Healthcare NHS Trust, Orpington, UK; South London Healthcare NHS Trust, Orpington, UK; South London Healthcare NHS Trust, Orpington, UK 63y.o lady presented with polyuria, polydipsia [amp] bony aches with elevated Corrected calcium 2.83mmol/l (2.02-2.6) PTH 243ng/l (15-65) and normal renal function. Vitamin D low: 13.4 mcg/l. Clinical examination was unremarkable. Further investigations revealed elevated 24 hour urinary calcium at 8.3mmol/24 hours (2.5-7.5). USS revealed suspicion of 4 gland disease (On the right side two lesions were seen posterior and inferior to the lower pole of the thyroid and on the left side, two further lesions again posterior and inferior to the lower pole were noted. The possibility of 4 gland hyperplasia was therefore raised from imaging). Sesta-MIBI scan showed bilateral inferior position uptake. DEXA scan revealed osteoporosis at spine with a T-score of -2.5 and osteopenia at hips with T-score of -1.7.[br]She underwent a total Parathyroidectomy given the imaging and histology showed 4 gland hyperplasia. Postoperatively bone mineral density improved. Genetic study was considered in view of 4 gland hyperplasia and was positive for mutations in MEN 1 gene. She had no history of peptic ulcer disease. There was no family history of endocrinopathies or hypercalcaemia. Further MEN 1 related tests were normal. Interestingly her daughter was referred with hypercalcaemia at time her mother was diagnosed with MEN 1. She herself tested positive for this mutation. The daughter[apos]s sesta MIBI scan showed uptake in right lower position. However given the family history and positive test for MEN 1 she is scheduled to have 3.5 gland parathroidectomy.[br]Conclusion: (1) Consider genetic studies in parathyroid hyperplasia [ndash]50% of cases are familial. (2) In MEN 1 positive cases, single uptake on sesta-MIBI does not necessarily rule out multiple gland hyperplasia and one need to be prepared to do a sub-total parathyroidectomy (or at least consider intra-operative PTH). (3) Imaging can help inform surgery.[br][br]Eur J Endocrinol September 1, 2008 159 259-27.[br][br]Nothing to Disclose: AS, PS, AT, AL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1054 188 1388 SUN-360 PO46-01 Sunday 1184 2012


1182 ENDO12L_SUN-361 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Pseudohypercalcemia [mdash] Uncommon Presentation of Multiple Myeloma Sujani Poonuru, Joseph L Shaker, Srividya Kidambi Medical College of Wisconsin, Milwaukee, WI [bold]Background: [/bold]Hypercalcemia is one of the presenting signs of multiple myeloma (MM). Increased bone resorption or bone destruction resulting in increased serum total and ionized calcium (iCa) is usually the primary etiology for hypercalcemia in patients with MM. Parathyroid hormone (PTH) is usually suppressed ([lt] 20 pg/mL) in these patients due to elevated iCa.[br][bold]Case:[/bold] A 65 year old male was referred to endocrinology clinic for increased serum calcium that was noted in his routine laboratory tests 3 months prior to presentation. The serum calcium one year ago was normal at 10.1 mg/dl. He had no known personal or family history of hypercalcemia. He reported fatigue and night sweats but was otherwise asymptomatic. His past history was significant for hypertension, for which he was on hydrochlorothiazide (HCTZ) 25 mg/day for the past 3 years. Physical examination was unremarkable. The laboratory tests revealed serum total calcium 11.5 mg/dL(8.5-10.1 mg/dl), albumin 2.1 g/dL(4-5.3 g/dL), albumin-corrected calcium 13.02 mg/dL, iCa 1.17 mmol/L (1.13- 1.32 mmol/L), phosphorous 2.8 mg/dL (2.5-4.9 mg/dL), creatinine 0.94 mg/dL (0.67-1.17 mg/dL), intact PTH 74.8 pg/mL (14-72 pg/mL), 25 (OH)D 33 ng/mL (30-100 ng/mL), 1,25 (OH2)D14 pg/mL (18-72 pg/mL), hematocrit 33.8% (40-51%). The presence of anemia and relatively rapid onset of hypercalcemia with normal iCa, raised the possibility of MM despite elevated PTH levels. Serum electrophoresis showed an IgG gammopathy 4.48 g/dL (0.5-1.25 g/dL) with kappa light chains. Radiographic studies revealed multiple bone lytic lesions. Bone marrow evaluation showed approximately 60% of cellularity is replaced by plasma cells, consistent with plasma cell myeloma. HCTZ was discontinued. The patient was treated with zoledronic acid which resulted in normalization of albumin corrected calcium, however, it lowered ionized calcium levels to below normal range.[br][bold]Discussion:[/bold] Here we present a rare phenomenon of pseudohypercalcemia as a presenting sign of MM. The increased total calcium is due to calcium binding to abnormal myeloma proteins, leading to lowering of iCa and subsequent elevation in PTH to keep iCa in normal range (1). It is also interesting to note that therapy with zoledronic acid in this setting can produce ionized hypocalcemia. Recognizing the existence of the phenomenon of pseudohypercalcemia can avoid mis-diagnosis of true hypercalcemia or primary hyperparathyroidism.[br][br]Jaffe JP, Mosher DF. Calcium binding by a myeloma protein. Am J Med. 1979;7:343-346.[br][br]Nothing to Disclose: SP, JLS, SK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 217 188 1389 SUN-361 PO46-01 Sunday 1185 2012


1183 ENDO12L_SUN-362 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Hypercalcemia Associated with Mineral Oil[ndash]Induced Sclerosing Paraffinomas Kerri Kissell, Andreas Moraitis, Martin Hewison, Michael Holick, Michael T Collins National Institutes of Health, Bethesda, MD; David Geffen School of Medicine at UCLA, Los Angeles, CA; Boston University School of Medicine, Boston, MA; National Institutes of Health, Bethesda, MD Background: Hypercalcemia occurs in a variety of chronic granulomatous diseases such as sarcoidosis, berylliosis and slack-skin granulomatous disease associated with lymphoma. The proposed mechanism involves increased circulating levels of dihydroxyvitamin D [1,25(OH)2D] catalyzed by 25 hydroxyvitamin D3 1-alpha hydroxylase (VD3 1A). This enzyme is known to be expressed by a variety of extrarenal cells, including macrophages and recently identified in subcutaneous adipose cells of neonates. We present a case of hypercalcemia secondary to extrarenal production of 1,25(OH)2D in a patient with extensive cutaneous and subcutaneous lesions associated with mineral oil injections. Clinical Case: A genotypic male was treated with oral estrogens for thirty years to achieve a phenotypic female appearance presents with recurrent symptomatic hypercalcemia. After ceasing oral estrogen use in 1997, the patient self-administered mineral oil injections in the buttocks, breast, and thighs from 1997-2000. Beginning in 1998, the patient had multiple skin and soft tissue lesions requiring surgical management and multiple hospitalizations for management of symptomatic hypercalcemia, with serum calcium levels ranging from 14 to 15.6 (8.9-10.1 mg/dL) and suppressed iPTH levels of [lt]7 pg/mL. There was no history of vitamin D supplementation, nephrolithiasis, or fracture. There was no family history of parathyroid or calcium disorders. At first presentation to our team, serum calcium: 15.6 mg/dL, phosphorous: 5.2 mg/dL, Cr: 2.0 mg/dL, Alkaline phosphatase: mildly elevated and unchanged since previous diagnosis of Hepatitis C. iPTH was suppressed at [lt]7.0 pg/mL. 24 hour urine calcium: 88 mg/24 hours with creatinine clearance of 45 ml/min. 25(OH)D: 7 ng/mL (20-100 ng/mL), 1,25 (OH)2D: 550 pg/mL (20-65 pg/mL), and PTHrP: 0.26 pmol/L ([lt]1.3 pmol/L). Biopsy was performed at edge of cutaneous and subcutaneous lesions involving thigh and buttocks, consistent with noncaseating granuloma. Immuno-localization of 1a-hydroxylase was identified within macrophages of the same specimen. Conclusions: Extensive skin lesions associated with mineral oil injections can precipitate hypercalcemia and elevated serum 1,25(OH)2D levels. Macrophages in the dermis and epidermis that express 1-hydroxylase are likely producing 1,25(OH)2D in an unregulated manner.[br][br]Nothing to Disclose: KK, AM, MH, MH, MTC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1667 188 1390 SUN-362 PO46-01 Sunday 1186 2012


1184 ENDO12L_SUN-363 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Hypercalcemic Crisis in a Pregnant Woman with Cervical Squamous Cell Carcinoma: Case Presentation and Literature Review Farnoosh Farrokhi, David Ziemer, Dawn Smiley, Francisco J Pasquel, Christopher Newton, Guillermo E Umpierrez Emory University School of Medicine, Atlanta, GA Background: Hypercalcemia during pregnancy is an uncommon but serious medical problem associated with a high rate of maternal/fetal demise. We report the first case of severe hypercalcemia of malignancy due to invasive squamous cell carcinoma (SCC) of the cervix during pregnancy.[br]Clinical Case: A 28 yo, G2P2 ptt, BMI 23 kg/m[sup]2[/sup] with 2 months of intermittent vaginal bleeding was found to have a high-grade squamous intraepithelial cervical lesion on pap smear and invasive SCC by colposcopy and biopsy. A PET-CT of the pelvis revealed an enlarged, heterogeneous uterus with FDG activity consistent with cervical cancer and bilateral enlarged iliac and obturator lymph nodes suggestive of stage IIB SCC. Upon admission she was found to be 5-weeks pregnant with a [beta]HCG of 52 mIU/ml and serum calcium of 16.5 mg/dl (nl: 8.9-10.3), albumin 3.4 gm/dl, and serum creatinine 1.5 mg/dl. After initial hydration, repeat calcium was 14.7 mg/dl, phosphorous 1.2mg/dl, serum PTH [lt] 6.0 pg/ml (nl: 12.0-88.0), PTH-related peptide 33 pg/ml (nl: 4-27), and a normal SPEP. Following the 24-48 hr administration of IVFs, her calcium level decreased to 12.2 mg/dl. Patient opted for pregnancy termination. Due to the high risk of bleeding from her cervical lesion, she underwent external pelvic radiation for cancer treatment and pregnancy termination. In addition to hydration, treatment of hypercalcemia included SQ calcitonin 0.4 u/kg q12 h for 2 days and IV pamidronate 90 mg. Four days later, her calcium level was normal (9.4 mg/dl) and her level remained within normal range at subsequent discharge while receiving external radiation therapy.[br]Conclusion: This is the first report of a patient with metastatic cervical SCC with severe hypercalcemia during pregnancy. A review of the literature revealed 11 cases of cervical SCC-associated hypercalcemia in non-pregnant women. Hypercalcemic crisis during pregnancy has devastating outcomes for both mother and fetus including hypertension, preeclampsia, pancreatitis, low birth weight, preterm delivery, seizures and neonatal death. Similar to nonpregnant patients, management includes forced hydration and bisphosphonate therapy. Few case reports of bisphosphonate therapy during pregnancy reported no teratogenic effects; however, these agents are classified as group D and during pregnancy may increase the risk of miscarriage and low fetal birth weight. We discuss diagnostic and treatment options for hypercalcemia of malignancy during pregnancy.[br][br]Nothing to Disclose: FF, DZ, DS, FJP, CN, GEU 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2108 188 1391 SUN-363 PO46-01 Sunday 1187 2012


1185 ENDO12L_SUN-364 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) A Case of Primary Hyperparathyroidism Due to a Parathyroid Adenoma, a Parathyroid Carcinoma and a Subsequent Adenoma May Uyking Naranjo St Luke[apos]s Medical Center, Quezon City, Philippines An extremely unusual course of primary hyperparathyroidism (PHP) due to a parathyroid adenoma in 2004, a carcinoma in 2010 and a subsequent adenoma in 2011 is presented herein. A 50-year-old man presented with chronic low back pain in 2004. On evaluation, he was found to have nephrolithiases. Further work up showed ionized calcium 1.54mmol/L(1.00-1.20), intact PTH 139pg/ml(7-53), crea 1.56mg/dL(0.60-1.3), GFR 52mL/min. He was diagnosed with PHP. A dual phase Tc99m sestamibi scan showed increased tracer uptake in the inferior pole of the right thyroid lobe. He was found to have a 3cm parathyroid adenoma of the right inferior gland. He remained normocalcemic until early 2010 he was hospitalized due to recurrent hypercalcemia with significantly high PTH level. Results: ionized Ca 1.38, intact PTH 4910, phosphorus 2.23mg/dl (2.69-4.49). Repeat parathyroid scan showed increased uptake in the inferior right thyroidal bed and the left thyroidal bed. No other abnormal accumulation of tracer in the neck or mediastinum was found. Radio-guided surgery and a histologic examination of the resected tumor provided evidence of a parathyroid carcinoma in the right inferior and superior gland (1gm and 2.3x1.2x0.8cm). Intra-operative PTH dropped from 4760 to 35.6. He developed hungry bone syndrome and was corrected with IV calcium and oral calcitriol.[br]In less than a year of normocalcemia, he was again hospitalized for recurrent hypercalcemia and hyperparathyroidism. A new lesion in the suprasternal notch area was found on repeat parathyroid scan. Metastatic parathyroid carcinoma was highly considered. Repeat radio-guided exploratory surgery was performed. The tumor was consistent with a parathyroid adenoma (1gram and 1.5x1.0x0.4cm) with central lymph node negative for metastasis. At greater than one year of follow-up, the patient had no evidence of disease recurrence and his calcium and PTH have remained within normal.[br]The course of parathyroid tumors are variable. PHP due to two recurrent bouts of parathyroid adenomas and a carcinoma occurring independently within a seven-year period is unusual. Parathyroid carcinoma may be suspected, but it usually cannot be confirmed prior to operation. Recurrent PHP after resection of a parathyroid carcinoma does not necessarily indicate a metastatic parathyroid carcinoma but can also be caused by an adenoma.[br][br]Nothing to Disclose: MUN 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 533 188 1392 SUN-364 PO46-01 Sunday 1188 2012


1186 ENDO12L_SUN-365 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Milk-Alkali Syndrome: A Re-Surfacing Cause of Hypercalcemia Ruban Dhaliwal, Pallawi Torka, Dinesh John, Jennifer Kelly, Arnold M Moses State University of New York (SUNY) Upstate Medical University, Syracuse, NY; State University of New York (SUNY) Upstate Medical University, Syracuse, NY [bold]Background:[/bold] There has been a resurgence of Milk-alkali syndrome in recent years, yet it remains underdiagnosed. Our case serves to heighten the awareness of this disorder as consumption of calcium has increased, now making it the third most common cause of hospitalization for hypercalcemia.[br][bold]Clinical case:[/bold] A 64 year-old male presented with diffuse abdominal pain, lethargy, constipation and recurrent falls due to profound weakness over last few days. Initial laboratory tests revealed serum calcium 21 mg/dL (8.5-10.2), creatinine 3.8 mg/dL, (0.7-1.4), phosphorus 3.4mg/dL (2.5-4.5), sodium 132 meq/L (135-145), potassium 2.9 meq/L (3.8-5.1), bicarbonate 35 meq/L (22-32). Albumin was low at 3.3 g/dL (3.4-4.5) and liver function tests were normal. EKG showed new onset Mobitz type 1 second degree heart block. Abdominal x-ray showed radioopaque stool in colon and several ingested tablets in the stomach. Medication history was reviewed. Patient had gastroesophageal reflux disease for several years with symptoms worsening despite use of sucralfate. This prompted him to take as many as 20 calcium carbonate tablets (Tums) daily. He had no prior history of hypercalcemia. Milk-alkali syndrome was diagnosed.[br]Aggressive hydration with normal saline resulted in decrease in calcium level to 18.4mg/dL within few hours. Subcutaneous calcitonin 4 units/kg twice a day was initiated. Calcium steadily decreased over next four days returning to normal (8.4 mg/dL). An upper GI endoscopy revealed esophagitis. Proton pump inhibitor was started. As hypercalcemia and alkalosis resolved, the patient[apos]s mentation improved, abdominal pain subsided and EKG normalized. Creatinine decreased to 2.3 mg/dL but did not return to baseline. PTH came back 31.9 pg/ml (8.5-72.5), 25-OH vitamin D was low at 11 ng/ml (30-100). Serum and urine protein electrophoresis were normal.[br][bold]Conclusion:[/bold] [apos]Modern[apos] milk-alkali syndrome, aptly renamed as [apos]Calcium-alkali syndrome[apos] is usually secondary to over-the-counter calcium carbonate intake, hence a detailed drug history is mandatory in hypercalcemia. Clinicians need to be aware of this entity due to its growing incidence. Patients typically present in acute hypercalcemic crisis with normal to low phosphate levels. Treatment involves withdrawal of offending drug and hydration. Bisphosphonates tend to cause hypocalcemia and should be avoided. Renal failure is multifactorial and residual renal deficit may be present even after resolution of hypercalcemia.[br][br]Nothing to Disclose: RD, PT, DJ, JK, AMM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1663 188 1393 SUN-365 PO46-01 Sunday 1189 2012


1187 ENDO12L_SUN-366 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Hyperparathyroidism (HPT) in a Teenager Due to a Solitary Adenoma Two Years after Radioactive Iodine (RAI) Therapy for Papillary Thyroid Carcinoma (PTC) Danielle L Gomez, Dorothy I Shulman, Wade Cressman, Irmel Ayala University of South Florida, St Petersburg, FL; All Children[apos]s Hospital, St Petersburg, FL; All Children[apos]s Hospital, St Petersburg, FL A 17 year-old male was found to have hypercalcemia (13.1 mg/dL) and hypophosphatemia (2 mg/dL) following admission for a transverse sinus thrombosis. He had a history of shunted hydrocephalus in infancy associated with a vein of Galen aneurysm, treated with coiling and embolization. At 15 years, he was diagnosed with PTC that was treated with a total thyroidectomy, neck dissection, RAI therapy, and L-thyroxine suppression. He received two doses of RAI a year apart, a total of 263mCi. Follow- up scan after the second therapy dose showed uptake in right neck. Three months after the second dose he developed headaches due to a thrombus in his transverse sinus. Serum calcium was 10 mg/dL, thyroglobulin (Tg) was [lt]1 ng/mL. He was treated with anticoagulation for one year. During the year, surveillance neck ultrasounds revealed stable appearance of a 1 cm nodule in the right thyroid bed. Fine needle aspirate was planned after stopping anticoagulation therapy, however, 1 week after stopping anticoagulation headaches acutely returned. Brain CT showed progressive, non-occlusive thrombosis of his transverse sinus. Hypercalcemia was identified (13.1 mg/dL, PTH 154 pg/ml) and anticoagulant therapy was resumed. Neck ultrasound and sestamibi scan revealed probable hyperfunctioning right parathyroid adenoma. Prior to proceeding with surgical excision, recombinant thyrotropin- stimulated Tg was [lt]1 ng/mL and total body I 131 scan was negative. The patient underwent removal of right parathyroid adenoma with resolution of hypercalcemia.[br]RAI is accepted treatment for Grave[apos]s disease and PTC. Since the 1980[apos]s there have been many reports of hyperparathyroidism following RAI treatment, occurring mostly in females, at an older age, and with an average latency period of 15- 20 years. The majority are adenomas in the thyroid bed. This case is unusual in that it occurred in a young male with latency period of only 2 years. Our patient had numerous CT scans during early and late childhood to evaluate his aneurysm/hydrocephalus status which increased his total body radiation exposure, possibly increasing his risk. We recommend periodic surveillance of serum calcium for all patients receiving RAI, including children.[br][br]Nothing to Disclose: DLG, DIS, WC, IA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2296 188 1394 SUN-366 PO46-01 Sunday 1190 2012


1188 ENDO12L_SUN-367 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Acute Hyperphosphatemia Induced by Intravenous Calcium Infusion in a Patient with Post-Surgical Hypoparathyroidism Edward A Rico, Owaise MY Mansuri, Carmel M Fratianni Southern Illinois University School of Medicine, Springfield, IL Acute hyperphosphatemia associated with intravenous (IV) Calcium infusion in hypoparathyroidism is underappreciated. We report such a case in a patient with longstanding post-surgical hypoparathyroidism post-IV calcium (Ca) infusion for symptomatic hypocalcemia. This 78-year-old woman had post-surgical hypoparathyroidism post-thyroidectomy [amp] was maintained at baseline on oral Ca without calcitriol. She was readmitted 2 weeks post-mitral valve replacement. She had been asymptomatic with respect to hypocalcemia during the prior admission with low normal range Ca 7.5-8.5 mg/dL (nl: 8.4-10.5) with sub-normal iPTH level 0.491 pmol/L [1.2-7.0] [amp] adequate 25-OH Vitamin D stores 35.2 ng/ml (nl:[gt]30). She was readmitted with nausea/anorexia [amp] symptomatic hypocalcemia with myoclonus [amp] prolonged QTc: Ca 6.4 mg/dL (nl:8.4-10.5); albumin 3.7 g/dL (nl: 3.4-4.9); ionized Ca 0.7 mmol/L(1.1-1.3) [amp] Mg 1.9 mg/dl (nl:1.8-2.6). With this hypocalcemic stimulus, iPTH level rose to low normal range 1.39 pmol/L [1.2-7.0]. A slow calcium gluconate IV infusion was administered with increase in Ca to 8.2 mg/dL [amp] ionized Ca to 1.0 mmol/L 12 hrs later. It is notable that prior phosphorus (Phos) levels had been between 2.8-4.7 mg/dL (nl: 2.2-4.5.) After Ca infusion, however, Phos rose progressively from 5.5, peaking at 8.5 mg/dL six hours post-infusion. During this period PO intake was poor [amp] renal function stable. While BUN 37 [amp] Creat 1.3 suggest chronic kidney disease, these were unchanged from prior. There was no secondary cause of this precipitous rise in Phos which we attribute to IV Ca infusion alone. To control serum Phos, dual therapy with both Sevelamer Carbonate [amp] Lanthanum Carbonate was needed. Both agents were stopped after 4 days as Phos decreased to 4.0 mg/dL. Although chronically this hypoparathyroid patient had serum iPTH levels below the normal reference range, she clearly had some residual parathyroid function since a modest iPTH response to a significant hypocalcemic stimulus was demonstrable. Phos resorption occurs mostly in the proximal renal tubule, where PTH-dependent adenylate cyclase is found. We hypothesize that the administration of IV Ca acutely suppressed PTH, thereby suppressing Phos excretion at NPT2 in the renal tubules with resultant acute hyperphosphatemia. Judicious monitoring of serum Phos [amp] aggressive use of phosphate binders are key to effective management of acute hyperphosphatemia associated with IV Ca infusion in postsurgical hypoparathyroidism.[br][br]Nothing to Disclose: EAR, OMYM, CMF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1051 188 1395 SUN-367 PO46-01 Sunday 1191 2012


1189 ENDO12L_SUN-368 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Severe Primary Hyperparathyroidism Presenting as Bilateral Femur Fractures with Widespread Brown Tumors Mimicking Bony Tumor Metastasis Kevin A Codorniz, Dana Haydel Loma Linda University, Loma Linda, CA; University of California, Irvine, Orange, CA [bold]Background:[/bold] Primary hyperparathyroidism is known more commonly today as an asymptomatic disease, found incidentally by a mild elevation of serum calcium on routine blood work. Prior to the routine measurement of serum calcium, this was not the case, and reports of severe boney disease were common.[br][bold]Purpose:[/bold] We present a historical and now atypical presentation of severe primary hyperparathyroidism.[br][bold]Case Presentation:[/bold] A 38 year old woman presented to the ED for severe bilateral lower extremity pain after a low-height, mechanical fall. X-rays revealed bilateral midshaft femur fractures with scattered lytic lesions. Initial labs were significant for serum calcium of 14mg/dl (8.5-10.2 mg/dl). Further imaging showed widespread lytic lesions which were thought to be due to metastasis. Further workup was significant for an intact PTH of 1639pg/ml (10-68 pg/ml) and serum protein electrophoresis with a IgA Kappa monoclonal protein present on immunofixation. Bone scan showed innumerable, diffuse osseous metastatic like lesions in the axial and appendicular skeleton. A biopsy taken at the time of surgical repair of her femur fractures was consistent with brown tumors of the bone. Neck ultrasound described a 3x1.6x1.6cm solid nodule in the right thyroid lobe and a sestamibi scan showed a focus of accumulation in this same region. The patient underwent successful removal of the lesion with a fall of intraoperative rapid PTH from baseline of 2228pg/ml to 50pg/ml upon removal. Histological examination revealed a benign parathyroid adenoma measuring 4.5cm in greatest dimension. The patient developed severe hungry bone syndrome post operatively, but had a good recovery. Six weeks postoperatively her PTH was 23 and her calcium was 9.5.[br][bold]Discussion: [/bold]The diagnosis of primary hyperparathyroidism (PHPT) in today[apos]s medical practice has generally been made earlier due in part to serum calcium becoming automated in the early 1970s. It is now rare to see advanced PHPT, especially in developed countries, where the most common presentation is asymptomatic hypercalcemia. Osteitis Fibrosa Cystica was the classical manifestation of PHPT but is now a very rare occurrence. Our patient had us revisit some of these historical presentations as she had severe boney disease with widespread brown tumors mimicking boney metastasis. This case also highlights some other atypical features of severe primary hyparathyroidism including the possible co-existence of monoclonal gammopathy.[br][br]Nothing to Disclose: KAC, DH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1481 188 1396 SUN-368 PO46-01 Sunday 1192 2012


1190 ENDO12L_SUN-369 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Recurrent Hypercalcemia in a Patient with Previous Parathyroid Surgeries Payal Satish Patel, Marco Marcelli, Sanjay Navin Mediwala Baylor College of Medicine, Houston, TX; Michael E DeBakey VA Medical Center, Houston, TX Introduction: Parathyroid carcinoma (PC) accounts for [lt]1% of cases of primary hyperparathyroidism. No single histopathologic feature is pathognomonic for PC thus diagnosis can be challenging. We present a case of recurrent hypercalcemia due to PC.[br]Clinical Case: A 66-year-old man presented after sustaining a right hip fracture from a low-impact fall and was found to have symptomatic hypercalcemia. He had a long-standing history of hypercalcemia with nephrolithiasis, necessitating left nephrectomy and was incidentally found to have renal cell carcinoma in 1994. He was eventually diagnosed with primary hyperparathyroidism in 2008 with I-PTH of 428 pg/dL (11-64 pg/dL) and had right inferior parathyroidectomy in 3/2010 with pathology showing parathyroid adenoma. Post-operatively, calcium levels normalized and I-PTH decreased to 8 pg/dL. He was subsequently lost to follow-up. I-PTH was now [gt]900 pg/dL with a corrected calcium of 14.2 mg/dL (8.4-10.2 mg/dL). Hypercalcemia was managed with calcitonin and bisphosphonates. Sestamibi scan did not show any hyperfunctioning parathyroid gland. Thyroid ultrasound revealed a 1.3cm vascular nodule compressing the right superior thyroid lobe. After hip surgery, patient underwent excision of right vascular nodule with pathology consistent with parathyroid carcinoma. Calcium normalized post-operatively but hypercalcemia reoccurred two weeks later. I-PTH was 258 pg/dL. Repeat thyroid ultrasound showed new right inferior and superior nodules suspicious for parathyroid neoplasms. He underwent a repeat neck dissection but no parathyroid tissue was found. Post-operatively, I-PTH was 231 pg/dL with a corrected calcium of 10.1 mg/dL and rising. Cinacalcet was started for management of hypercalcemia.[br]Conclusion: Surgery is the first line of therapy for suspected PC; radiation and chemotherapy have poor results. Close follow-up is needed post-operatively as recurrence for PC is high. Hypercalcemia is the main cause of morbidity and mortality from PC and must be medically managed when PC is no longer amenable to surgery.[br]Furthermore, mutations in HRPT2 tumor suppressor gene (also called CDC73) has been implicated in sporadic as well as a familial form of hyperparathyroidism- the hyperparathyroidism-jaw tumor syndrome (HPT-JT). HPT-JT patients are predisposed to jaw fibromas, uterine tumors and renal neoplasms. This patient interestingly also had renal cell carcinoma and is undergoing genetic evaluation for HRPT2 mutations.[br][br]Nothing to Disclose: PSP, MM, SNM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1115 188 1397 SUN-369 PO46-01 Sunday 1193 2012


1191 ENDO12L_SUN-370 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Resolution of Hypercalcemia Following Correction of Vitamin D Deficiency: A Distinct Phenotype of Primary Hyperparathyroidism (PHPT)? Ogoma M Obi, Abdul Abou-Samra Wayne State University/Detroit Medical Center, Detroit, MI A 53 year old asymptomatic African American female presented with hypercalcemia (11.4 mg/dL), high PTH (320 pg/mL), high 24 h urine calcium (500 mg/day) and severe Vitamin D deficiency (25OHD [lt] 5 ng/mL). She had hypothyroidism due to radioactive iodine ablation for Graves[apos] disease, hypertension, hereditary spherocytosis, polycystic ovarian syndrome and a benign thyroid nodule. She had no fractures or renal stones and her family history was negative for parathyroid disorders. Her medications included levothyroxine, amlodipine and ramipril. Repeat testing confirmed PHPT (Ca of 11.5 mg/dL and PTH of 236 pg/mL). Considering the high PTH and marked hypercalciuria, Vitamin D was cautiously replaced with cholecalciferol at 1000 IU daily and frequent monitoring of serum Ca. Follow up at 6 weeks revealed serum Ca of 11 mg/dL with 25OHD of 10 ng/mL. At 3 months, her serum calcium was normalized (9.3 mg/dL); 25OHD was 17 ng/mL. DXA scan revealed osteopenia of the lumbar spine but a normal BMD of the forearm and hip. Sestamibi nuclear scan showed evidence of a parathyroid adenoma in the right lower thyroid lobe. The neck ultrasound showed stable nodular changes of the thyroid. At this time, it was felt that the patient did not meet the criteria for parathyroid surgery any more ([underline]1[/underline]). Alendronate was started for osteopenia and cholecalciferol was continued. At 11 months (the most recent visit) serum calcium was 9.9 mg/dL, 25OHD was 20 ng/mL and PTH remained elevated at 221 pg/mL. Her 24 hour urine calcium excretion was 458 mg/day.[br][bold][underline]Discussion[/underline][/bold]: PHPT is relatively common, characterized by PTH mediated hypercalcemia ([underline]2[/underline]). Vitamin D deficiency is also prevalent, frequently coexisting with PHPT ([underline]3[/underline]) and may worsen the clinical manifestations of PHPT ([underline]4[/underline], [underline]5[/underline]). Vitamin D supplementation often unmasks latent PHPT. Current guidelines recommend screening PHPT patients for vitamin D deficiency and repletion if 25OHD levels [lt] 20 ng/mL ([underline]2[/underline]). This is often done cautiously to avoid worsening of hypercalemia and hypercalciuria ([underline]3[/underline], [underline]4[/underline]) though Ca levels remain stable in most patients.[br]Our patient demonstrated resolution of hypercalcemia following vitamin D repletion. This raised several questions: Does severe prolonged vitamin D deficiency result in parathyroid hyperplasia which ultimately triggers adenoma formation ([underline]6[/underline])? Is there a spectrum in the pathogenesis of primary hyperparathyroidism with an [bold][apos]early[apos][/bold] stage where treatment of inciting factors (e.g. vit D deficiency) reverses the phenotype?[br][br]1. Bollerslev J, Marcocci C, Sosa M, Nordenstrom J, Bouillon R, Mosekilde L 2011 Current evidence for recommendation of surgery, medical treatment and vitamin D repletion in mild primary hyperparathyroidism. European journal of endocrinology / European Federation of Endocrine Societies 165:851-864. 2. Eastell R, Arnold A, Brandi ML, Brown EM, D[apos]Amour P, Hanley DA, Rao DS, Rubin MR, Goltzman D, Silverberg SJ, Marx SJ, Peacock M, Mosekilde L, Bouillon R, Lewiecki EM 2009 Diagnosis of asymptomatic primary hyperparathyroidism: proceedings of the third international workshop. The Journal of clinical endocrinology and metabolism 94:340-350. 3. Jithpratuck W, Garrett LH, Peiris AN 2011 Treating vitamin D insufficiency in primary hyperparathyroidism: a cautionary tale. Tennessee medicine : journal of the Tennessee Medical Association 104:47-49. 4. Grey A, Lucas J, Horne A, Gamble G, Davidson JS, Reid IR 2005 Vitamin D repletion in patients with primary hyperparathyroidism and coexistent vitamin D insufficiency. The Journal of clinical endocrinology and metabolism 90:2122-2126. 5. Silverberg SJ 2007 Vitamin D deficiency and primary hyperparathyroidism. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research 22 Suppl 2:V100-104. 6. Mallya SM, Gallagher JJ, Wild YK, Kifor O, Costa-Guda J, Saucier K, Brown EM, Arnold A 2005 Abnormal parathyroid cell proliferation precedes biochemical abnormalities in a mouse model of primary hyperparathyroidism. Mol Endocrinol 19:2603-2609.[br][br]Nothing to Disclose: OMO, AA-S 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 872 188 1398 SUN-370 PO46-01 Sunday 1194 2012


1192 ENDO12L_SUN-371 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Hypercalcemia: More Than the Usual Hope Torregosa, Sathya Krishnasamy, Ali Cahid Civelek, Bryan K Moffett University of Louisville, Louisville, KY; University of Louisville, Louisville, KY; Robley Rex Veterans Affairs Medical Center, Louisville, KY Primary hyperparathyroidism may be aymptomatic or manifest with nephrolithiasis, bone disease, neuromuscular or neuropsychiatric symptoms. Although often mentioned in the literature, few cases of primary hyperparathyroidism have presented as acute pancreatitis. We present a case of recurrent pancreatitis due to an ectopic parathyroid adenoma.[br]A 54-year old Caucasian man with obesity, coronary artery disease requiring multiple percutaneous coronary interventions, type 2 diabetes mellitus, hyperlipidemia and gastroesophageal reflux disease presented with acute pancreatitis for the second time in six months. Etiology could not be explained by hepatobiliary disease, hypertriglyceridemia, alcohol, medications, infections or anatomical variants. Aside from hypercalcemia (corrected calcium 10.7 and 11.0 mg/dL; normal 8.4-10.2 mg/dL) at presentation on both occasions, a review of his laboratory history revealed mild hypercalcemia (10.4-11.0 mg/dl) for at least 8 years. He denied nephrolithiasis, osteopenia or fractures. The parathyroid hormone level was was normal at 31 pg/ml with 24-hour urine calcium of 300 mg. Phosphorus and 25-hydroxy vitamin D levels were both normal. An ectopic retromanubrial parathyroid adenoma was suggested on a tomographic SPECT/CT sestamibi scan. After recovering from pancreatitis, he underwent minimally invasive radio-guided parathyroid surgery two months later, at which time a 2.2 gram adenoma was removed from the retromanubrial area. The serum calcium level has remained normal since surgery with no recurrence of pancreatitis.[br]The gastrointestinal implications of primary hyperparathyroidism are often overlooked until symptoms arise. Gastric acid hypersecretion is associated with peptic ulcer disease, while gastrointestinal atony leads to constipation, abdominal pain, nausea and decreased appetite. Deposition of calcium ions causes pancreatic ductal obstruction and inflammation. Hypercalcemia may also promote the conversion of trypsinogen to trypsin. Prompt recognition of hypercalcemia as a main cause of pancreatitis can result in appropriate treatment, in this case, via removal of a parathyroid adenoma to prevent recurrence of pancreatitis.[br][br]Nothing to Disclose: HT, SK, ACC, BKM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2345 188 1399 SUN-371 PO46-01 Sunday 1195 2012


1193 ENDO12L_SUN-372 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Pre- and Post-Surgery Evolution of Calcium, Phosphorus and Bone Remodeling Markers in Severe Primary Hyperparathyroidism Philippe Caron, Pierre D[apos]Amour, Philippe Caron CHUM, Montreal, Canada 34 years old woman from Cameroun was evaluated for severe symptomatic primary hyperparathyroidism with a hot bone scan and subperiosteal bone resorption. Her initial 25 (OH) D was low at 12 (N = 75-150) with an Intact (I) PTH of 119 (N =1.4-6.8), an ionized calcium of 1.92 (N = 1.16-1.29), a total calcium of 3.38 (N = 2.17-2.56), an alkaline phosphatase of 720 (N = 36-110) and a C-Telopeptide of 0.670. Pre-surgery, the patient was treated with 20 000 units of vitamin D a week for two weeks. She also received a dose of 60 mg of pamidronate and aggressive IV hydratation. After this treatment, her Intact (I) PTH rose to 144.5, her C-telopeptide rose to 2.37, her 25(OH) D rose to 67 while her serum ionized calcium dropped to 1.46 and her alkaline phosphatase dropped to 386. Both the parathyroid scan and the echography identified a left inferior adenoma which was benign at pathology and weighted 1.6g. After the surgery the patient was treated with 20 000 units of cholecalciferol per week and 1000 mg of calcium a day. Despite this treatment, the patient developed hypocalcemia (ionized calcium 1.13) and secondary hyperparathyroisim (I-PTH 35.3). It took 4 months to normalize her calcium and her PTH. This patient illustrates importance of pre and post surgery vitamin D therapy for primary hyperparathyroidism.[br][br]Rankin W, Aust Fam Phys 2011;881-4. Bollerslev J, Europ J Endocrinol 2011; 851.[br][br]Nothing to Disclose: PC, PD, PC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 766 188 1400 SUN-372 PO46-01 Sunday 1196 2012


1194 ENDO12L_SUN-373 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Alendronate and Vitamin D Replacement in Asymptomatic Tertiary Hyperparathyroidism Mei Zhu, Xiangzhuo Zhao, Baoping Wang, Zhihong Gao Tianjin Medical University General Hospital, Tianjin, China [bold]Background:[/bold] Tertiary hyperparathyroidism (TPHT), which was the result of a long-standing, severe secondary hyperparathyroidism, has turned autonomous after the renal transplantation. As an alternative to surgery, alendronate and vitamin D were prescribed to reduce serum calcium level and increase serum level of 25-hydroxyvitamin D in asymptomatic TPHT.[br][bold]Clinical case: [/bold]A 46-year-old woman was referred to our department with five years history of persistently increased PTH level, and three years of asymptomatic hypercalcaemia after renal transplantation. The patient had been clinically diagnosed with purpuric nephrosis, uremia with increased PTH of 18.5 pmol/L (normal: 1.1-7.8) five years before. Three years before, she underwent renal transplantation. One month after the surgery, her serum test showed: calcium 2.67 mmol/L (normal: 2.15-2.55), phosphate 0.42 mmol/L (normal: 0.8-1.45) and PTH 21.5 pmol/L. However, the patient lacked specific symptoms traditionally associated with hypercalcemia or PTH excess. She was monitored every three months. Her serum calcium fluctuated in 2.15 to 2.80mmol/L, and PTH was persistently elevated (15.7-24.0 pmol/L). An increased 24 hr urinary calcium excretion (350.4mg) was found in the absence of kidney stones. Creatinine clearance was 67 ml/min. Measurement of bone mineral density by DEXA showed osteopenia at both lumbar spine (T-score -1.6) and femoral neck (T-score -2.1). Ultrasound showed hypoechoic mass below the lower pole of the right lobe of thyroid gland. These findings indicated the diagnosis of tertiary hyperparathyroidism. Surgery was not recommended because of patient preference and the indications for surgery were not met in this asymptomatic THPT patient. As an alternative to surgery, alendronate was prescribed to the patient in order to increase bone density. Her 25-hydroxyvitamin D was 14.58 nmol/l (normal 47.7-144) and 1,25(OH)[sub]2[/sub]VitD[sub]3[/sub] was 10.13pmol/l (normal 39-193), which implied co-existing vitamin D deficiency in this patient. Therefore, vitamin D supplementation to this patient targeted to increase vitamin D concentration. Unexpectedly, decreased in both serum calcium (2.35mmol/L) and 24 hr urinary calcium excretion (288.6mg) were found.[br][bold]Conclusion: [/bold]Alendronate in cases of hyperparathyroidism unable to undergo surgery, gives the opportunity to improve the vitamin D status of the patient without aggravating hypercalcemia.[br][br]Sources of Research Support: National Natural Science Foundation of China(30973040).[br][br]Nothing to Disclose: MZ, XZ, BW, ZG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 522 188 1401 SUN-373 PO46-01 Sunday 1197 2012


1195 ENDO12L_SUN-374 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Coexisting Severe Vitamin D Deficiency and Primary Hyperparathyroidism: A Suggested Management Strategy Sridevi Ganapathy, Adrian Hugh Heald, Mohammad Zubair Qureshi Leighton Hospital, Crewe, UK; The University of Manchester, Manchester, UK The management of hypercalcemia secondary to primary hyperparathyroidism is challenging where there is severe coexisting vitamin D deficiency. We describe such a case here.[br]A 62 year female with a history of osteoarthritis and hypertension was admitted with lethargy and arthralgia but no history of renal calculi or relevant family history. Drugs included bendroflumethiazide, candesartan, and NSAIDS. Initial investigations were serum adjusted calcium 3.4 mmol/L (normal range(nr) 2.2-2.6), phosphate 0.61 mmol/L (nr 0.8-1.5), PTH 54.3 pmol/L (nr 1.3-6.8) and creatinine 87 [micro]mol/L (nr 50-100). Severe vitamin D deficiency was identified (Vitamin D2 [lt]4[micro]g/L, vitamin D3 [lt]6[micro]g/l). Serum magnesium levels was within the normal range and coeliac screen was negative. Renal ultrasound were normal and urinary calcium excretion 3.6mmol/24 hours. Bone mineral densitometry measurements revealed non-dominant forearm density in the osteopaenic range (T score -2.3) compared to hip (T score -1) and spine (T score -1.9).[br]Adequate fluid rehydration was administered and bendroflumethiazide discontinued. Vitamin D replacement was not undertaken at this stage due to the risk of worsening hypercalcemia. USS neck and Sestamibi imaging were compatible with a left inferior parathyroid adenoma compatible with primary hyperparathyroidism. Continued symptomatic severe hypercalcemia required frequent intravenous hydration pending parathyroidectomy.[br]A single oral dose of 200,000 IU of vitamin D was administered a day prior to parathyroidectomy to decrease risk of post-operative severe hypocalcemia and excess calcium resorption in bones ([apos]hungry bone syndrome[apos]).Histology confirmed benign parathyroid adenoma.[br]An 8-week course of oral ergocalciferol 1.25mg once per week (50,000 IU) was also prescribed. With this serum vitamin D and calcium levels returned to normal (adjusted calcium 2.32 mmol/L, phosphate 1.2 mmol/L, creatinine 84 mmol/L, TSH 1.96 mU/L, 25(OH)D2 79.6 nmol/L, 25(OH)D3 18.6 nmol/L). PTH concentrations were marginally raised at 6.9 pmol/L (normal 1.3- 6.8). Significant symptomatic improvement was observed.[br]Vitamin D replacement can worsen hypercalcemia, while vitamin D deficiency increases the risk of the [apos]hungry bone[apos] syndrome after parathyroidectomy with adverse consequences. We suggest that in severe vitamin D deficiency co-existing with severe hypercalcemia in primary hyperparathyroidism, replacement of vitamin D in the immediate peri-operative period is a sensible strategy.[br][br]Nothing to Disclose: SG, AHH, MZQ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1480 188 1402 SUN-374 PO46-01 Sunday 1198 2012


1196 ENDO12L_SUN-375 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) A Rare Case of Hypercalcemia Vallikantha Nellaiappan, Gokulakrishnan Balasubramanian, Preethi C Sridhar, Balasubramanian Rathinasabapathy Rosalind Franklin University of Medicine and Science, North Chicago, IL; Charukrishna Clinic, Chennai, India Background: Primary Hyperparathyroidism is often asymptomatic. Even the symptoms are subtle and often missed initially. Ectopic parathyroid is a common reason for failure to locate parathyroid glands during surgery.[br]Clinical Case: A 53 year old gentleman presented with myalgia, severe leg cramps, abdominal pain and loss of appetite of 15 days duration. He had non bloody vomiting and constipation for 3 days. Past medical history was significant for type 2 diabetes mellitus of 3 years duration. Vital signs and physical examination were unremarkable. Lab work showed serum glucose of 225mg/dl(N:80-140), BUN 64.3 mg/dl(N:13-45), creatinine 2.45mg/dl(N:0.7-1.2), elevated calcium 20mg/dl(N:8.6-10.2) and low phosphate of 2.1mg/dl(N:4-7). Further testing showed a TSH of 3.1mcg/ml(N:0.2-4.5) and PTH of 1593pg/ml(15-65). CT neck and upper chest showed well defined retrosternal nodule of 1.3cm X 2.5cm while CT abdomen was normal. Technetium (99mTc) sestamibi scan showed abnormal focus of activity in substernal region and caudal to the lower pole of left lobe of thyroid gland. Renal function and calcium level improved with intravenous hydration, subcutaneous calcitonin and followed by oral cinacalcet. Patient underwent surgical removal of the ectopic parathyroid. Histopathology was consistent with parathyroid adenoma. His calcium and PTH were within normal limits (8.6mg/dl and 24.62pg/ml respectively) after the surgery.[br]Discussion: Primary hyperparathyroidism is equally common among young men and women while preferentially affects women in aged [1]. Estimated incidence as per Rochester Epidemiological Project is 22 cases per 100,000 persons per year[2]. Most cases of primary hyperparathyroidism are due to parathyroid adenoma, majority of which occurs in neck. 10% of parathyroid adenomas are ectopic, of which 50% occur in anterior mediastinum[3]. Usually they are encapsulated and [lt] 3cm in size. Chest X ray and CT scan are not sensitive modality for its diagnosis. Technetium (99mTc) sestamibi scan has a sensitivity of 88-100% in detecting these lesions[3]. Combining 2 imaging studies such as CT chest and neck with Technetium (99mTc) sestamibi scan improves the sensitivity and gives valuable anatomical information before surgery [4]. For symptomatic primary hyperparathyroidism, surgery is the treatment of choice[5].[br][br]1. Marcocci, C. and F. Cetani, Clinical practice. Primary hyperparathyroidism. N Engl J Med, 2011. 365(25): p. 2389-97. 2. Wermers, R.A., et al., Incidence of primary hyperparathyroidism in Rochester, Minnesota, 1993-2001: an update on the changing epidemiology of the disease. J Bone Miner Res, 2006. 21(1): p. 171-7. 3. Strollo, D.C., M.L. Rosado de Christenson, and J.R. Jett, Primary mediastinal tumors. Part 1: tumors of the anterior mediastinum. Chest, 1997. 112(2): p. 511-22. 4. Lumachi, F., et al., Technetium-99m sestamibi scintigraphy and helical CT together in patients with primary hyperparathyroidism: a prospective clinical study. Br J Radiol, 2004. 77(914): p. 100-3. 5. Fraker, D.L., Update on the management of parathyroid tumors. Curr Opin Oncol, 2000. 12(1): p. 41-8.[br][br]Nothing to Disclose: VN, GB, PCS, BR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1591 188 1403 SUN-375 PO46-01 Sunday 1199 2012


1197 ENDO12L_SUN-376 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Hypercalcemia Associated with Silicone-Induced Granulomas in Male-to-Female Transsexual Patients Karen Racedo, So-Young Kim, Carla Romero, Amit Seth, David Shipman, Alina Gouller Beth Israel Medical Center, New York, NY; Beth Israel Medical Center, New York, NY [bold][italic]Objective:[/italic][/bold][br]To report cases of severe hypercalcemia in male-to-female (MTF) transsexual patients due to subcutaneous (SQ) liquid silicone injections (SI).[br][bold]Case 1[/bold]: 41yo MTF transsexual HIV patient with h/o multiple SI into hips since 1989, presented with polyuria, polydipsia, lethargy [amp] nodules over both hips. Labs showed elevated calcium (Ca) (13mg/dl, nl[lt]10.3), ACE (85U/L, nl[lt]67); normal creatinine (Cr), 1,25-OH vitamin D (1,25OHD) (47pg/ml, nl[lt]75pg/ml) [amp] PTH-rp ([lt]2pmol/L, nl[lt]4); low 25-OH vitamin D (25OHD) (16pg/ml, nl[gt]30) [amp] PTH ([lt]3pg/ml, nl[gt]11). CT scan showed nodular expansion of gluteal tissues and enlarged inguinal lymph nodes (LN). LN biopsy revealed reactive hyperplasia with lipogranulomas. Patient was treated with IV steroids [amp] fluids with normalization of Ca (9.9mg/dl). She was discharged on prednisone, but was non-complaint and re-admitted with Ca of 18.4mg/dl. After IV fluids, steroids, and pamidronate, Ca normalized and she was discharged on prednisone and alendronate. At 4 month visit, Ca was normal.[br][bold]Case 2[/bold]: 31yo MTF transsexual HIV patient with h/o SI into both hips since 1993 presented with lethargy, vomiting, polyuria, polidipsia and SQ nodules over hips [amp] calves. Labs showed elevated Ca (14.7mg/dl), ACE (207U/L); low PTH ([lt]3pg/ml), PTH-rp ([lt]2.5pmol/L) and 25OHD (10pg/ml); normal 1,25OHD (50 pg/ml), Cr, SPEP and UPEP. CT scan revealed SQ nodules and prominent LN. LN biopsy showed reactive hyperplasia with lipogranulomas. Sarcoid was ruled out. After IV fluids [amp] pamidronate, Ca normalized and she was discharged on alendronate. SQ nodules were debulked for cosmetic reasons, but Ca remained elevated. She is maintained on alendronate and intermittent steroids.[br][bold][italic]Discussion:[/italic][/bold][br]We report persistent hypercalcemia in transsexual patients due to liquid SI. Silicone can migrate through tissue, invade muscles and cause granulomas [amp] lymphadenopathy.[sup]1[/sup] A proposed mechanism for high Ca is activation of macrophages that accelerates conversion of 25OHD to 1,25OHD.[sup]2[/sup] 1,25OHD promotes Ca absorption, bone resorption and decreases Ca excretion. Both patients had suppressed PTH, low 25OHD, inappropriately normal 1,25OHD (in setting of high Ca), supporting the proposed mechanism of hypercalcemia.[br][bold][italic]Conclusion:[/italic][/bold][br]Siliconomas and hypercalcemia are late complications of SI. Hypercalcemia can be successfully treated with bisphosphonates and steroids. Siliconomas are unresectable. In our case, debulking did not resolve hypercalcemia but can be considered for refractory cases.[sup]3[/sup][br][br](1) Caskey CI, Berg WA, Hamper UM, Sheth S, Chang, BW, Anderson ND. Imaging spectrum of extracapsular silicone: correlation of US, MR imaging, mammographic, and histopathologic findings. Radiographics. 1999;19(Spec No):S39-S51. (2) Mason RS, Frankel T, Chan YL, Lissner D, Posen S. Vitamin D conversion by sarcoid lymph node homogenate. Ann intern med 1984; 100:59-61. (3) Khan O, Sim JJ. Silicone-induced Granulomas and Renal Failure. Dialysis [amp] Transplantation 2010; 39 (6): 254-259.[br][br]Nothing to Disclose: KR, S-YK, CR, AS, DS, AG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2354 188 1404 SUN-376 PO46-01 Sunday 1200 2012


1198 ENDO12L_SUN-377 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) Inactive Sarcoidosis Unmasked by an Acute Presentation of Hypercalcemia Secondary to Milk-Alkali Syndrome Shadi Abdelnour, Morali D Sharma University of Nevada School of Medicine, Las Vegas, NV; Baylor College of Medicine, Houston, TX [bold]Background[/bold]: Hypercalcemia is a common complication in milk-alkali syndrome (MAS) and sarcoidosis (SAR). We report a case of hypercalcemia, where treatment for MAS partially corrected the calcium (Ca) and creatinine (Cr), but full recovery was only achieved after prednisone was initiated. [bold]Clinical Case[/bold]: A 70-year-old Caucasian male with a history of SAR diagnosed in 1957, and treated with glucocorticoids for 6 months who remained asymptomatic with normal Ca and Cr levels thereafter. He presented with fatigue, decreased appetite, 10-lb weight loss, occasional nausea, polyuria and polydipsia. His physical exam was unremarkable. He reported daily intake of 2 liters of milk for more than 20 years, and several Ca tablets for the past 1 year to relieve his epigastric pain and GERD symptoms. Work-up revealed the following: Ca 13.4 mg/dL(8.5-10.5), ionized Ca 1.58 mmol/L(1.12-1.27), albumin 4.1 g/dL(3.5-5), phosphorus 4 mg/dL(2.5-4.5), BUN 40 mg/dL(5-26), carbon dioxide(CO2) 33 mm/L(20-32), Cr 3.3 mg/dL(0.76-1.27), intact PTH 11 pg/mL(7-80), PTHrP 18 pg/mL(14-27), 25-hydroxy vitamin D[25(OH)D] 18 ng/mL(20-100), 1,25-dihydroxy vitamin D[1,25(OH)2D] 82 pg/mL(18-72), and ACE(angiotensin-converting enzyme) 102 U/L(9-67). Other tests including sodium, potassium, magnesium, cortisol, CBC, SPEP, UPEP, US of the kidneys and CT of the abdomen and pelvis were unremarkable, but his chest CT revealed bilateral reticular markings within the lung bases. Pulmonary function tests showed mild decrease in diffusion capacity. The patient responded well to intravenous fluids and was discharged home. Two weeks later, without any Ca or milk intake his labs revealed Ca 12.1 mg/dL, albumin 4.2 g/dL, Cr 1.59 mg/dL, BUN 23 mg/dL, CO2 23 mm/L, 1,25(OH)2D 75 pm/L, and ACE 114 U/L. The patient was started on prednisone 40 mg PO daily for two weeks and the repeat Ca was 9.6 mg/dL with Cr of 1.1 mg/dL. The patient did not have any ophthalmic or dermatological findings of SAR. Hypercalcemia recurred as soon as the prednisone was discontinued, so prednisone was resumed at 10 mg PO daily, which controlled his hypercalcemia. [bold]Conclusion[/bold]: SAR can present as hypercalcemia despite the absence of common clinical features. The presence of normal baseline Ca and Cr levels, the distant history of SAR, the high level of 1,25(OH)2D in the presence of renal failure, the ACE level and the response to treatment with prednisone are likely consistent with inactive SAR which was unmasked by the MAS.[br][br]Nothing to Disclose: SA, MDS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 62 188 1405 SUN-377 PO46-01 Sunday 1201 2012


1199 ENDO12L_SUN-378 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) A New Deletion in [italic]CYP24A1[/italic] in Patients with Idiopathic Infantile Hypercalcemia Matthieu Decamp, Nadia Coudray, Genevieve Abeguile, Nicolas Richard, Herve Mittre, Nicolas Gruchy, Eric Mallet, Marie-Laure Kottler CHU de Caen, Caen, France; CHU de Rouen, Rouen, France Background: CYP24A1 catalyzes the conversion of 25-hydroxyvitamin D3 and 1,25- hydroxyvitamin D3 into 24-hydroxylated products, the first step of vitamin D degradation. Lack of CYP24A1 activity leads to hypercalcemia encountered in idiopathic infantile hypercalcemia (IIH)[sup]a[/sup].[br]Objective: The aim of the present study was to screen patients with IIH for [italic]CYP24A1[/italic] gene mutations.[br]Patients: Index case was a male patient who was followed since the age of 7 months for typical severe IIH. He presented with failure to thrive after failing to gain any weight in the preceding 3 months. We found high serum calcium level (3.68 mmol/l), high 1,25-dihydroxyvitamin D3 level and low PTH level. He had a short QT interval on ECG and a bilateral nephrocalcinosis. His twin brother and his parents were normal; however his second brother displays the same feature.[br]Methods: We performed PCR amplification of the 11 exons of [italic]CYP24A1[/italic] gene. The non-PCR amplification of exons 9 to 11 of the [italic]CYP24A1[/italic] gene for the index case promted us to test the DNA for Copy Number Variant using a TaqMan[reg] Copy Number Variation Assay. Each family member (parents, his non-affected twin brother and his affected brother) were tested. Two CNV assays were performed, one in intron 8 to exon 9 junction and the other in intron 10 to exon 11.[br]Results: We confirm the lack of amplification of exons 9 to 11 of the [italic]CYP24A1[/italic] gene in the two affected brothers, in which we found an homozygous deletion. The two deleted alleles were inherited from each heterozygous parents.[br]Conclusion: We described for the first time a partial deletion of [italic]CYP24A1[/italic] gene in family with IIH encompassing exon 9 to exon 11. The size of the deletion is under investigations. This new deletion underlies the role of [italic]CYP24A1[/italic] mutations in IIH.[br][br]Nothing to Disclose: MD, NC, GA, NR, HM, NG, EM, M-LK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1855 188 1406 SUN-378 PO46-01 Sunday 1202 2012


1200 ENDO12L_SUN-379 POSTER SESSION: Bone [amp] Calciotropic Hormones (1:30 PM-3:30 PM) How Much Is Too Much: A Case of Hypercalcemia Geeteshwar Mangat, Pranav Ghody, Amolika Mangat, Zohirul Islam, Patricia Park State University of New York (SUNY) Downstate at Long Island College Hospital, Brooklyn, NY The fraction of the U.S. population consuming at least one dietary supplement has jumped from 42% to 53% in the last decade. Contrary to traditional belief, excess consumption of vitamins can have significant detrimental effects. We describe an unusual case of hypercalcemia associated with excess ingestion of over the counter calcium and vitamin D supplements.[br]A 54 year old woman with no prior medical history presented with symptoms of headache, myalgias, weakness and vomiting along with photophobia and subjective fevers for 10 days. She denied having abdominal pains or shortness of breath. On questioning, the patient admitted consuming [gt] 20 different vitamin supplements ([gt]75 pills/day) including multiple formulations of calcium and vitamin, for the last 5 years. Physical exam was significant for right upper quadrant abdominal tenderness without rebound or guarding. Admission labs were significant for Calcium 14.5 mg/dL (normal: 8.5-10.5); Phosphorus 6.3 mg/dL (normal 2.5-4.5); Vitamin 25(OH) D 295 ng/mL (normal: 30-80); undetectable Intact PTH, and BUN 60 mg/dL; Creatinine 2.3 g/dL;PTH Related Protein less than 2.0 mL; normal thyroid hormones and unremarkable serum and urine protein electrophoresis. Hypercalcemia was treated aggressively with intravenous saline and the patient[apos]s renal function improved with normalization of calcium levels. The patient was determined to have hypercalcemia from excess supplementation and was discharged home with outpatient follow up and instructions to avoid interim supplements of calcium and vitamin D.[br]The Institute of Medicine recommends that for females age 51- 70 years, calcium and vitamin D requirements are 1200 mg/day and 600 IU/day respectively. Our patient had been consuming 2605.2 mg of elemental calcium and more than 4000 IU of vitamin D per day for several years, well above the recommended limits.[br]Excess ingestion of vitamin D and calcium can cause hypercalcemia which can present with both acute and chronic systemic toxicities. Studies show that most patients do not report the use of alternative medications to their physicians. However, when evaluating a patient, it is very important to note the consumption pattern and constituents of OTC supplements and herbal medicines. In the absence of FDA regulation of herbal medications, the exact concentrations of constituents may be unknown or differ from the labels. Ingestion of such products may lead to side effects and create a diagnostic dilemma for physicians.[br][br]Nothing to Disclose: GM, PG, AM, ZI, PP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1824 188 1407 SUN-379 PO46-01 Sunday 1203 2012


1201 ENDO12L_SUN-380 POSTER SESSION: Other Metabolic Bone Diseases (1:30 PM-3:30 PM) Osteosclerosis in Congenital Hypophosphatemic Bone Disorders Saket Gupta, Mark Kilbane, Susan Vanderkamp, Malachi John McKenna St Vincent[apos]s University Hospital, Dublin, Ireland; St Vincent[apos]s University Hospital, Dublin, Ireland; St Vincent[apos]s University Hospital, Dublin, Ireland; St Vincent[apos]s University Hospital, Dublin, Ireland Congenital renal phosphate wasting encompasses inheritable disorders that present as rickets during childhood and osteomalacia during adulthood. The most common form is X-linked hypophosphataemia (XLH) due to loss of function mutations of the phosphate-regulating gene with homology to endopeptidases located on the X chromosome (PHEX). Rarer forms included autosomal dominant hypophosphataemic rickets (ADHR) due to mutation in FGF23 gene, and autosomal recessive hypophosphataemic rickets (ARHR) due to loss of function of dentin matrix protein. Despite having a mineralisation defect, bone mineral density (BMD) is not reduced and may be increased in XLH. Although, this matter is neither mentioned nor addressed in a recent review (1).[br]We sought to determine the BMD by dual-energy X-ray absorptiometry (DXA) at spine and hip in 11 patients: 9 with XLH, 1 with ADHR, and 1 without any of the known mutations. The age range was 19[ndash]59 years (mean[plusmn]SD: 33.4[plusmn]10.6 years). There were 10 females and 1 male. Most of them were on various treatment regimens for XLH. Osteosclerosis was defined as a BMD Z-score [ge]2.0 above the mean for age-and-sex-matched reference range. Seven of 11 had osteosclerosis at the spine; 3 of 11 had osteosclerosis at the hip all of whom had high BMD at spine. 3 patients had osteosclerosis at hip at time of initial DXA examination. A 33 years old female was followed up for 11 years who did not respond to standard treatment, underwent total parathyroidectomy and subsquent DXA revealed osteosclerosis at both spine and hip (spine Z-score = 5.5; hip Z-score = 3.3). The patient with ADHR was the only patient with Z-score below the mean: spine Z-score = -0.4; hip Z-score = -0.6. The eldest patient, a 60-year-old man, had a Z-score at spine of 8.8. At the most recent examination the mean Z-scores (mean[plusmn]SD) for all 11 patients were as follows: spine BMD, 3.2[plusmn]2.4, hip BMD, 1.1[plusmn]1.5. The total bone mineral densities at spine and hip were 1.4[plusmn]0.3gm/cm2 and 1.1[plusmn]0.2 gm/cm2 respectively. Spine bone mineral measurements were significantly greater than those for hip bone mineral measurements *p[lt]0.05.[br]Osteosclerosis is a neglected but common finding in XLH that appears to increase with age. Our data demonstrate that BMD may be discordant with a tendency toward decreased BMD toward extremities and increased BMD in the lumbar spine consistent with previous studies (2). We suggest that BMD should be monitored at least every 2 years in adult patients with XLH.[br][br](1) Carpenter et al., J Bone Mineral Res 2011;26:1381-1388. (2) Rosenthall et al., Clin Nucl Med 1993; 18:564[ndash]566.[br][br]Nothing to Disclose: SG, MK, SV, MJM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1753 189 1408 SUN-380 PO05-02 Sunday 1204 2012


1202 ENDO12L_SUN-381 POSTER SESSION: Other Metabolic Bone Diseases (1:30 PM-3:30 PM) The Effect of Calcium Supplementation on FGF23 Concentration Vickie Braithwaite, Inez Schoenmakers, Ann Prentice Elsie Widdowson Laboratories, Cambridge, UK; MRC Laboratories, Lower River Division, Gambia Fibroblast growth factor-23 (FGF23) is a phosphate (PO[sub]4[/sub]) regulating hormone in conjunction with 1,25-dihydroxyvitamin D (1,25(OH)[sub]2[/sub]D) and parathyroid hormone (PTH). Additionally, 1,25(OH)[sub]2[/sub]D and PTH are both primary regulators of plasma calcium (Ca). It is therefore plausible that FGF23 is modulated by dietary Ca intake. However, studies in mice and humans have shown conflicting effects of Ca supplementation on FGF23 concentration[sup](1, 2)[/sup].[br]This study tests whether Ca supplementation in children with an habitually low dietary Ca ([sim]200 mg/d)[sup](3)[/sup], affects FGF23 concentration. We measured FGF23 concentration in plasma samples from a subset of Gambian children ([italic]n[/italic]=40) who participated in a randomized, placebo-controlled Ca supplementation trial (ISRCTN28836000)[sup](4,5)[/sup]. They received either 2x500 mg of elemental Ca (CaCO[sub]3[/sub]) (S; [italic]n[/italic]=19) or placebo (P; [italic]n[/italic]=21) 5 x/week for 1 year. Fasting blood samples and 24 h urine were collected before and after 1 year of supplementation.[br]C-terminal plasma FGF23 was measured by ELISA (Immutopics Inc., USA), PTH and 1,25(OH)[sub]2[/sub]D by immunoradiometric assay (Diasorin, Ltd, UK) and plasma ([italic]p[/italic]) and urine ([italic]u[/italic]) Ca, PO[sub]4[/sub] and Cr by colorimetric methods. Data are presented as mean (SD) or geometric mean (-SD, +SD).[br]FGF23 concentrations at baseline and after a year of supplementation were: S= 62.4 (35.2, 110.9) and 60.0 (36.4, 98.9) and P= 70.6 (27.8, 179.5) and 64.3 (32.1, 129.3) RU/ml. Using regression analysis, and correcting for baseline FGF23, sex and age, the change ([Delta]) in FGF23 was not different between groups ([italic]P[/italic]=0.7). In keeping with the results from the main trial, PTH in the subset decreased by 5.9 (SD 9.4) pg/ml in S but increased by 4.5 (SD10.4) pg/ml in P. [Delta]PTH was different between groups (S-P [Delta]%=-29.5, [italic]P[/italic]=0.006). 1,25(OH)[sub]2[/sub]D decreased in both groups but more substantially in S (31.7 (SD35.6) pmol/l) than in P (6.3 (SD29.3) pmol/l). [Delta]1,25(OH)[sub]2[/sub]D was different between groups (S-P [Delta]%=-15.8, [italic]P[/italic]=0.002). There was no difference in [Delta][italic]p[/italic]Ca and [Delta][italic]p[/italic]PO[sub]4[/sub] between groups. Supplementation increased [italic]u[/italic]Ca:[italic]u[/italic]Cr and [Delta][italic]u[/italic]Ca:[italic]u[/italic]Cr was different between groups (S-P [Delta]%=81.6, [italic]P[/italic]=0.003). [italic]u[/italic]PO[sub]4[/sub]:[italic]u[/italic]Cr decreased in both S and P but more so in S and [Delta][italic]u[/italic]PO[sub]4[/sub]:[italic]u[/italic]Cr was different between groups (S-P [Delta]%=-47.5, [italic]P[/italic]=0.0009).[br]The results indicate that Ca supplementation in children with an habitually low dietary Ca intake had little effect on C-FGF23 concentrations despite having measurable effects on its regulators.[br][br](1) Shimada T, et al., Am J Physiol Renal Physiol 2004; 289:F1088. (2) Khadilkar A, et al., Arch Dis Child 2009; 94:932. (3) Braithwaite V, et al., Bone 2012; 50:218. (4) Dibba B, et al., Am J Clin Nutr 2000; 71:544. (5) Dibba B, et al., Am J Clin Nutr 2002; 76:681.[br][br]Sources of Research Support: This study was supported by the UK Medical Research Council [Unit Programme numbers U105960371 and U123261351].[br][br]Nothing to Disclose: VB, IS, AP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 471 189 1409 SUN-381 PO05-02 Sunday 1205 2012


1203 ENDO12L_SUN-382 POSTER SESSION: Other Metabolic Bone Diseases (1:30 PM-3:30 PM) Hypophosphatasia: Importance of Low Alkaline Phosphatase and Fracture as Markers for Disease in Adults Michael P Whyte, Cheryl R Greenberg, Priya S Kishnani, Laura Case, Jill Mayhew, Catherine Sly Seiner, Nerissa C Kreher, Hal Landy, Vivienne T Lim Shriners Hospital for Children, St Louis, MO; University of Manitoba, Winnipeg, Canada; Duke University Medical Center, Durham, NC; Duke University Medical Center, Durham, NC; Shriners Hospital for Children, St Louis, MO; Washington University School of Medicine, St Louis, MO; Enobia Pharms, Cambridge, MA; Shriners Hospital for Children, St Louis, MO Background: Hypophosphatasia (HPP) results from inactivating mutation(s) in the gene for the tissue non-specific isoenzyme of alkaline phosphatase (ALP). In perinatal and infantile HPP, symptoms and signs are profound and include significant skeletal hypomineralization, failure to thrive, long bone fractures, rickets, nephrocalcinosis, respiratory compromise, vitamin B-6 dependent seizures, and/or premature loss of teeth. In juvenile (childhood) and adult HPP, the clinical features may be subtle or nonspecific but morbidity is significant. The diagnosis can be missed if serum ALP is not measured as low ALP is the hallmark of HPP.[br]Objective: To assess the disease burden experienced in adults with HPP[br]Methods: Two surveys, one via the internet (The HPP Impact Patient Survey [HIPS], 9/2009[ndash]6/2011) and one via telephone interview (The HPP Outcomes Study Telephone [HOST], 12/2010[mdash]3/2011), explored the impact of HPP on children and adults (i.e., [gt] 18 yr). 184 patients (59 children and 125 adults) completed surveys. Volunteer and potential study subjects were self-selected (HIPS), or were referred by specialists in metabolic bone disease at medical centers (HOST).[br]Appropriate informed consent or assent was obtained. Here we report the results of surveys in 125 adult patients. A subset of those 125 adults had their disease diagnosed at [ge] 18 yrs and are defined as [apos]adult-onset[apos] HPP.[br]Results: Among the 125 adults surveyed ([ge] 18 yrs when surveyed), there were 44 subjects who were diagnosed as [apos]adult-onset[apos] disease; however, as many as 25% of these [apos]adult-onset[apos] patients actually reported HPP manifestations in childhood. The medical history of adult-onset patients revealed 100% experienced bone or joint pain. 98% had experienced at least one fracture in their past with 64% of patients requiring surgical fixation of fracture. The first symptoms or signs reported in these adult-onset HPP patients were fracture (40%), pain (20%) and tooth loss (17%). In response to their disease, 32% modified their homes and 27% employed help for activities of daily living.[br]Conclusion: Even in patients diagnosed with [apos]adult-onset[apos] HPP, morbidity and disability are substantial. Understanding the signs and symptoms of HPP in children and adults, with low ALP activity being the hallmark of this disease, is pivotal to making this diagnosis.[br][br]Disclosures: MPW: Principal Investigator, Enobia Pharmaceutical; Speaker, Enobia Pharmaceutical. CRG: Principal Investigator, Enobia Pharmaceutical. PSK: Principal Investigator, Endo Pharmaceuticals Inc. LC: Researcher, Endo Pharmaceuticals Inc. JM: Consultant, Enobia Pharmaceutical. CSS: Collaborator, Enobia Pharmaceutical. NCK: Employee, Endo Pharmaceuticals Inc. HL: Employee, Endo Pharmaceuticals Inc. VTL: Collaborator, Enobia Pharmaceutical. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2314 189 1410 SUN-382 PO05-02 Sunday 1206 2012


1204 ENDO12L_SUN-383 POSTER SESSION: Other Metabolic Bone Diseases (1:30 PM-3:30 PM) A Typical Acute Oral Calcium Load Has No Adverse Effect on Vascular Compliance and Endothelium Reactivity Assessed Non-Invasively in Healthy Subjects Marianna Yaron, Vanessa Roach, Elena Izkhakov, Maya Ish-Shalom, Jessica Sack, Yael Sofer, Ibrahim Azzam, Assaf Ray, Naftali Stern, Karen M Tordjman Tel Aviv Sourasky Medical Center, Tel Aviv, Israel [bold]Background:[/bold] Calcium supplementation is a mainstay of osteoporosis prevention and treatment. However, recent studies have implicated calcium supplementation as a potential cause for an increased incidence of cardiovascular events among older women (1). It was proposed that transient elevations of serum calcium could have an adverse effect on vascular function. Furthermore, it was suggested that, coming from food, calcium might be devoid of such an effect, as it is typically accompanied by far milder perturbations of serum calcium (2). The aim of this study was to non-invasively assess vascular function in response to an acute calcium load in young, healthy subjects.[br][bold]Methods:[/bold] Eleven healthy, vitamin D-sufficient, volunteers (8 females and 3 males) aged 33 [plusmn] 6.1 years, BMI 22.6[plusmn] 2.3 kg/m[sup]2[/sup], participated in this random-order, cross over study. Biochemical, hormonal, and vascular parameters before and 3h after a typical acute oral calcium (600 mg) load, administered either as calcium citrate or as non-fat dairy products, were compared. Arterial stiffness was studied non-invasively by measuring Pulse wave velocity (PWV), Augmentation index (Alx) and Large (C1) and Small (C2) arterial compliance. Endothelial function was assessed by reactive hyperemia-induced flow mediated dilation (FMD). Finally, intimal-media thickness was measured once. As each subject served as her/his own control, paired t-test or paired ANOVA were used to analyze the results.[br][bold]Results:[/bold] The acute calcium supplement raised serum calcium from 9.1[plusmn] 0.3 to 9.6 [plusmn] 0.4 mg/dl, P[lt]0.005, while the same amount coming from food resulted in an insignificant increase from 9.1[plusmn]0.5 to 9.3[plusmn]0.3 mg/dl. The degree of urinary calcium excretion (expressed as Ca/Cr) was similar after both challenges. Moreover, PTH was equally and significantly suppressed: from 29.9[plusmn]11.2 to 13.6[plusmn]7.4 pg/ml for the supplement (P[lt]0.001), and from 31.8[plusmn]11.5 to 12.6[plusmn]6.6 for the food study (P[lt]0.001). Despite clear biochemical evidence for effective calcium loading on both occasions, none of the vascular parameters were affected by either challenge.[br][bold]Conclusions[/bold]: An acute oral calcium load appears to have no appreciable untoward effect on the vascular properties of young healthy subjects, regardless of the way it is provided. Additional studies will attempt to determine if these findings are applicable to post-menopausal women.[br][br](1) Bolland MJ et al., BMJ 2008; 336:262. 92) Reid IR et al., Clin Endorinol 2010; 73:689.[br][br]Nothing to Disclose: MY, VR, EI, MI-S, JS, YS, IA, AR, NS, KMT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 427 189 1411 SUN-383 PO05-02 Sunday 1207 2012


1205 ENDO12L_SUN-384 POSTER SESSION: Other Metabolic Bone Diseases (1:30 PM-3:30 PM) Hyperinsulinemia and Urinary Calcium Excretion in Calcium Stone Formers vs. Non-Stone Formers Vivienne Yoon, Naim M Maalouf, Khashayar Sakhaee University of Texas Southwestern Medical Center, Dallas, TX [bold]Background:[/bold] Several metabolic studies have shown an exaggerated rise in postprandial urine calcium excretion in hypercalciuric calcium stone formers compared to non-stone formers. It is not known if insulin or glucose mediate this postprandial hypercalciuria. We sought to examine the effects of hyperinsulinemia independent of glucose on urinary calcium excretion during a 2-hour hyperinsulinemic euglycemic clamp.[br][bold]Methods:[/bold] 22 non-stone formers and 7 hypercalciuric calcium stone formers were recruited across a range of body mass index. Fasting serum and 2-hour urine were collected prior to and during the clamp procedure.[br][bold]Results:[/bold] Twenty-two non-stone formers (9 male/13 female) with an average age (47 [plusmn] 12 years) and body mass index (26 [plusmn] 6 kg/m[sup]2[/sup]) completed the clamp. Seven hypercalciuric calcium stone formers (5 male/2 female) with an average age (49 [plusmn] 7 years) and BMI (28 [plusmn] 6 kg/m[sup]2[/sup]) completed the clamp. A baseline 24 hour urine collected the day prior to the clamp showed calcium stone formers had significantly higher urine calcium compared to non-stone formers (240 [plusmn] 57 mg/24hr vs. 166 [plusmn] 65 mg/24hr, p=0.01). There was no change in serum calcium concentration in calcium stone formers (8.9[plusmn] 0.4 vs. 8.9[plusmn] 0.3 mg/dL, p=0.8), and non-stone formers (8.8 [plusmn] 0.3 vs. 8.8 [plusmn] 0.3 mg/dL, p=0.5) before and during the clamp. There was a significant rise in fractional excretion of calcium (FeCa) from baseline to hyperinsulinemia (2.1 [plusmn] 1.1 vs. 2.7[plusmn] 1.2%, p=0.002) in non-stone formers. However, there was no significant rise in FeCa from baseline to hyperinsulinemia (2.2 [plusmn] 1.4 vs. 2.4 [plusmn] 1.5%, p=0.6) in calcium stone formers.[br][bold]Conclusions: [/bold]Hyperinsulinemia increases urine calcium excretion independent of changes in serum glucose in non-stone formers, and may potentiate calcium stone risk in this population. However, hyperinsulinemia does not significantly increase urine calcium excretion in hypercalciuric calcium stone formers. Therefore, hyperinsulinemia does not play a major role in calcium stone risk in hypercalciuric calcium stone formers.[br][br]Nothing to Disclose: VY, NMM, KS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1731 189 1412 SUN-384 PO05-02 Sunday 1208 2012


1206 ENDO12L_SUN-385 POSTER SESSION: Other Metabolic Bone Diseases (1:30 PM-3:30 PM) Therapy with Disodium Pamidronate in Children with Osteogenesis Imperfecta [mdash] A Single Center Experience Mirjana Kocova, Elena Sukarova-Angelovska University Pediatric Clinic, Skopje, Macedonia Osteogenesis imperfecta (OI) is a hereditary disorder due to the decreased amount or abnormal structure of the collagen. Major symptoms of the disease are bone pain and multiple fractures that lead to the significant physical handicap.[br]Bisphosphonates given i.v. improve the bone density, and ameliorate the symptoms, decreasing the rate of bone fractures in children of all age groups. However, no precise guidelines exist, and different studies show different outcomes (1,2).[br]The aim of our study was to evaluate the effect of treatment with bisphosphonates (disodium pamidronate) in children with osteogenesis imperfecta in a single clinical center.[br]Children with OI were diagnosed according the standard criteria (multiple fractures caused by minor trauma, phenotype, family history, X-ray, DEXA, alkaline phosphatase). Quality of life questionnaire was completed by the parents in 14 children after at least 1 year of treatment, and by the patients older than 11 years (N=10).[br]Seventeen children with all forms of OI were included in the study. Their age at the onset of treatment was: 1 month-13 years (4.55 years average). Age at the assessment was 3-16 years. The total number of fractures in all children before diagnosis was 78 (3.9 + 2.1 per child average). All patients were treated with disodium pamidronate, in a dose 6.0-10.1 mg/kg/year. During the first 6 months the therapy was given monthly, at two months interval until a year, 4 times of 2 day-cycles per year during the second year, and twice 2 day-cycles yearly afterwards.[br]Clinical changes were evaluated regularly during treatment, and radiological changes were assessed yearly. During treatment of 1-11.2 years (4.17+ 2.5 years average) the deviation from normal bone mineral density (BMD) evaluated by DEXA, as indicated by the z-score, diminished from [minus]6.8 [plusmn] 1.1 to[minus] 2.8 [plusmn] 2.1 (P [lt] 0.001). Febrile episode appeared in 7 of the patients after the first dose. No other side effects were noticed.[br]The total number of fractures (11) during the follow up period decreased sevenfold (p[lt]0.0001). However, one severe fracture of the femur required roding. The quality of life of patients and their families also increased significantly.[br]In conclusion, i.v. treatment with disodium pamidronate in children with osteogenesis imperfecta improves BMD, decreases significantly number of fractures, and prevents severe deformities. Impact of the bisphosphonates on the bone structure requires further studies.[br][br]1. Glorieuh FH, Icholas DN, Ishop OP et al.Cyclic administration of pamidronate in children with severe osteogenesis imperfect a N Engl J Med 339(14):947-952. 2. Forlino A, Cabral WA, Barnes AM [amp] Marini JC New perspectives on osteogenesis imperfect Nature Rev Endocrinology 2011;7: 540-557 | doi:10.1038/nrendo.2011.81.[br][br]Nothing to Disclose: MK, ES-A 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1401 189 1413 SUN-385 PO05-02 Sunday 1209 2012


1207 ENDO12L_SUN-386 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) The Prevalence of Antithyroid Drug Use during Pregnancy among Women with Live Births in Northern California Joan C Lo, Scott A Rivkees, Malini Chandra, James Korelitz, Michael W Kuzniewicz Kaiser Permanente Northern California, Oakland, CA; University of Florida College of Medicine, Gainesville, FL; Westat, Rockville, MD BACKGROUND: Graves[apos] disease accounts for the majority of cases of hyperthyroidism among women of reproductive age. Pharmacologic treatment includes methimazole (MMI) or propylthiouracil (PTU), but contemporary population-based studies of the prevalence of antithyroid drug (ATD) use during pregnancy are lacking.[br]METHODS: We utilized data from Kaiser Permanente Northern California (KPNC), a large integrated healthcare delivery system, to ascertain live births occurring between January 1996 and December 2010 among women age 15-49 years old. Estimated date of conception (EDC) was based on gestational weeks at delivery. Electronic pharmacy records were examined for prescription of ATD between the EDC and delivery date to determine ATD exposure during pregnancy. The prevalence of ATD use during pregnancy was examined overall and across specific race/ethnic subgroups.[br]RESULTS: We identified 454,360 live births from 1996 to 2010, ascertaining the first birth for each woman per calendar year. Among these women, the average age was 29.7 [plusmn] 6.0 years and 42.9% were White, 7.4% Black, 25.2% Hispanic, 19.7% Asian and 4.8% other or unknown race. A total of 588 women (129 per 100,000 births) received an ATD prescription (average maternal age of 31.3 [plusmn] 5.7 years and 26.4% [ge] age 35 years at delivery). Most received only PTU (86.6%) while the remainder received PTU and MMI (8.3%) or MMI alone (5.1%) during their pregnancy. Nearly all ATD-treated women (96.4%) had a diagnosis of thyrotoxicosis before, during or after pregnancy. Among all live births, the annual prevalence of ATD exposure ranged from 95-159 per 100,000 births across the 15-year period with no specific temporal trend. The frequency of ATD prescribing varied significantly by race/ethnicity (p[lt] 0.01), with the highest frequency (per 100,000) among Asians (225) and Blacks (182), followed by Whites (105) and Hispanics (88).[br]CONCLUSION: Within a large diverse population of Northern California women experiencing live births, the overall prevalence of prescribed ATD use during pregnancy averaged 129 per 100,000 births over 15 years and varied by race/ethnicity. Most women received PTU, in accord with standard recommendations during pregnancy. Given new guidelines advising MMI for non-pregnant women due to PTU hepatotoxicity, future studies should examine the relative maternal-fetal risks of PTU and MMI to optimize pharmacologic management of hyperthyroidism during pregnancy.[br][br]Sources of Research Support: NIH Grant HD065200 awarded to SAR.[br][br]Nothing to Disclose: JCL, SAR, MC, JK, MWK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1703 190 1414 SUN-386 PO39-01 Sunday 1210 2012


1208 ENDO12L_SUN-387 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Breastmilk Iodine Concentrations Following Acute Dietary Iodine Intake Angela M Leung, Lewis E Braverman, Xuemei He, Timothy Heeren, Elizabeth N Pearce Boston University School of Medicine, Boston, MA; Boston University School of Public Health, Boston, MA [bold]Background: [/bold]Breastmilk iodine levels may vary temporally with recent dietary iodine intake. Breastmilk iodine levels may also be decreased by environmental exposure to perchlorate (ClO4, a ubiquitous natural substance and industrial contaminant) and thiocyanate (SCN, from the diet and cigarette smoke), competitive inhibitors of the sodium/iodide symporter (NIS) present in lactating breast cells. ClO4 and SCN may impair infants[apos] iodine availability from breastmilk or, due to its presence in breastmilk, inhibit infants[apos] thyroidal NIS to directly affect infant thyroid function. We assessed the effect of and time to peak breastmilk iodine levels after iodine ingestion. [bold]Methods:[/bold] Sixteen healthy lactating Boston-area women with no known thyroid disease were each given 600 [mu]g oral potassium iodide (KI) (456 [micro]g iodine) after an overnight fast. Levels of iodine, ClO4, and SCN were measured in breastmilk and urine samples obtained at baseline and hourly for 8 hours following iodine intake. All dietary iodine ingested during the study period was measured. [bold]Results: [/bold]Mean age of mothers was 30.2 [plusmn] 4.1 (SD) years. One mother smoked cigarettes and 2 were exposed regularly to secondhand smoke. Dietary iodine sources provided an additional 36-685 [micro]g of iodine intake during the 8-hour study period. Median (interquartile range) baseline iodine levels were 45.5 (34.5-169.0) [micro]g/L in breastmilk and 67.5 (57.5-140.0) [micro]g/L in urine. Following 600 [micro]g KI administration, the median increase in breastmilk iodine levels above baseline was 280.5 (71.5-338.0) [micro]g/L, and the median peak breastmilk iodine concentration was 354 (315-495) [micro]g/L. The median time to peak breastmilk iodine levels following KI administration was 6 (5-7) hours. Over the 8 hours, there were no significant changes in median breastmilk ClO4 (4.6 [3.2-6.4] [micro]g/L) and SCN (206 [84.5-319.5] [micro]g/L) levels. [bold]Conclusions:[/bold] Following ingestion of 600 [micro]g KI, there is a measurable rise in breastmilk iodine concentrations, with peak levels occurring at 6 hours. These findings strongly suggest that breastmilk iodine concentrations should be interpreted in relation to recent iodine intake. Breastmilk ClO4 and SCN concentrations remained stable during the 8 hours of the study period and were not correlated with breastmilk iodine levels.[br][br]Sources of Research Support: NIH/NICHD 1 K23 HD068552 01 (Leung); Boston University Clinical [amp] Translational Science Institute 1UL1RR025771.[br][br]Nothing to Disclose: AML, LEB, XH, TH, ENP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 323 190 1415 SUN-387 PO39-01 Sunday 1211 2012


1209 ENDO12L_SUN-388 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) A Nationwide Survey on the Seasonality of Iodine Intake and Iodine-Deficiency Status in Latvia Ilze Konrade, Maija Dambrova, Marina Makrecka, Lolita Neimane, Ieva Strele, Edgars Liepins, Aivars Lejnieks, Parsla Vevere, Ugis Gruntmanis, Valdis Pirags Riga Stradin[scaron] University, Riga, Latvia; Latvian Institute of Organic Synthesis, Riga, Latvia; Children[apos]s Clinical University Hospital, Riga, Latvia; University of Texas, Southwestern Medical Center and North Texas Veterans Affairs Medical Center, Dallas, TX; University of Latvia, Riga, Latvia [bold]Background:[/bold] Previous nation-wide survey on iodine deficiency in Latvia was conducted 10 years ago. The aim of the follow-up study was to determine the prevalence of iodine deficiency in spring and autumn months among schoolchildren and to analyse the neonatal thyrotropin (TSH) screening data.[br][bold]Design and methods[/bold]: A cross-sectional interseasonal school-based cluster survey of 915 children aged 9[ndash]12 from 46 randomly selected schools in all regions of Latvia. The urine samples were collected in October 2010 and April 2011. The prevalence of TSH[gt]5 mIU/L from the Latvian Neonatal TSH screening was also investigated.[br][bold]Results:[/bold] The median creatinine-standardised urinary iodine (UIC) concentration was 107.3 (IQR 69.1-161.7) [mu]g/g. Lower median UIC was detected during the spring sampling (78.3 [mu]g/g Cr) in comparison to the autumn sampling (129.7 [mu]g/g Cr). UIC measurements indicative of mild iodine deficiency were present in 31.6%, moderate deficiency in 11.9% and severe deficiency in 2.8% of participants. Based on the result averages, a significant difference in comparison to autumn data was established in the spring data, for which an IUC below 100 [mu]g/g was present in 63% of schoolchildren and an IUC below 50 [mu]g/g in 28.2% of schoolchildren.[br]The prevalence of TSH[gt]5 mIU/L was 8.2% in 2009 and 9.3% in 2010, which indicates a mild iodine deficiency. The frequency of elevated neonatal TSH in April was markedly higher than in September (8.3% vs. 4.1%, p [lt] 0.001).[br][bold]Conclusion:[/bold] In the absence of a mandatory salt iodisation program, there is a persisting iodine deficiency among Latvian schoolchildren during the spring season. The data from the National Neonatal TSH Registry confirm seasonal differences in iodine levels and iodine deficiency in Latvia.[br][br]Sources of Research Support: Latvian National Research Program in Biomedicine and Public Health 2009-2013.[br][br]Nothing to Disclose: IK, MD, MM, LN, IS, EL, AL, PV, UG, VP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1420 190 1416 SUN-388 PO39-01 Sunday 1212 2012


1210 ENDO12L_SUN-389 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Fecundity in Young Adults Treated Early for Congenital Hypothyroidism Is Related to the Initial Severity of the Disease: A Longitudinal Population-Based Cohort Study Yasmine Hassani, Beatrice Larroque, Sophie Dos Santos, Emmanuel Ecosse, Jean Bouyer, Juliane Leger Inserm U1018, Bic[ecirc]tre, France; Assistance Publique-H[ocirc]pitaux de Paris, H[ocirc]pital Beaujon, Clichy, France; Assistance Publique-H[ocirc]pitaux de Paris, H[ocirc]pital Robert Debr[eacute], Paris, France Untreated hypothyroidism is known to impair fecundity. Patients with congenital hypothyroidism born before the introduction of neonatal screening suffered from severe brain damage and few went on to have children. It is generally assumed that patients treated for CH do not suffer from such consequences of the disease for fecundity. However, it has only recently become possible to test this assumption, because screening programs have only been running for the last 30 years.[br]The aim of this study was to compare the fecundity of young adults treated early in the CH program with that of a reference group and to determine whether fecundity was related to disease severity or quality of treatment.[br][bold]Patients and Methods: [/bold]Of the 1748 subjects diagnosed with CH in the first 10 years after the introduction of neonatal screening in France (1978[ndash]1988), 1158 completed a questionnaire on fecundity at a mean age of 25.3 years. This self-administered questionnaire focused on first attempts to have a child and time-to-pregnancy. The reference group was that used in an analogous study on subjects born in France between 1971 and 1985. Fecundability hazard ratios were estimated with Cox regression models and adjusted for known fecundity confounders (age, smoking, reproductive history).[br][bold]Results:[/bold] Fecundability was similar for the CH and control groups: HR = 1.14 [0.89-1.47] for women and HR = 0.98 [0.58-1.66] for men. In women, the most severe initial forms of the disease [ndash] athyreosis, absence of bone maturation at the knee epiphyseal ossification centers and a low serum free T4 concentration at diagnosis ([lt]5 pmol/liter) [ndash] were associated with lower fecundity: HR=0.68 [0.50-0.98], (p=0.02); HR=0.65 [0.45-0.94], ([italic]p[/italic]=0.02) and HR=0.70 [0.50-0.97], ([italic]p[/italic]=0.03), respectively. However, fecundability was not associated with age at the start of treatment, initial levothyroxine dose or the adequacy of hypothyroidism control during childhood and young adulthood.[br][bold]In conclusion,[/bold] this study provides no evidence for the general impairment of fecundity in patients treated early for CH, who are now adults. However, fecundity was found to be lower in women suffering from the most severe form of the disease. Our results suggest that these women present mild subfertility. These results have important clinical implications, as the investigation of ovarian function should be offered to this group of patients. Further studies are also required, to explore the physiopathological mechanism involved.[br][br]Sources of Research Support: The French Ministry of Health (Programme Hospitalier de Recherche Clinique AOM 05011) and by the Wyeth Foundation for Children and Adolescents.[br][br]Nothing to Disclose: YH, BL, SdS, EE, JB, JL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 384 190 1417 SUN-389 PO39-01 Sunday 1213 2012


1211 ENDO12L_SUN-390 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Etiology of Increasing Incidence of Congenital Hypothyroidism in New Zealand from 1993 to 2010 Benjamin B Albert, Craig Jefferies, Dianne Webster, Wayne S Cutfield, Alistair J Gunn, Joan Carll, Kathy Bendikson, Jose G Derraik, Paul L Hofman University of Auckland, and Starship Children[apos]s Health, Auckland, New Zealand; Starship Children[apos]s Health, Auckland, New Zealand; Auckland District Health Board, Auckland, New Zealand; University of Auckland, Auckland, New Zealand; Ministry of Health, Auckland, New Zealand [bold]Background[/bold][br]Recent reports suggest that the incidence of congenital hypothyroidism (CHT) may be increasing in the USA and Western Australia. The aetiology of this change is unclear and it may in part be related to changes in screening thresholds. The possibility of an environmental cause for increasing rates has been raised. Thyroid scintiscan of newly identified CHT cases allows categorisation into thyroid dysgenesis (including athyreosis and ectopic thyroid), considered sporadic but with possible environmental influence, or dyshormonogenesis, an autosomal recessive disease. Thus routine scintiscan may help to identify the cause of changing incidence of CHT.[br][bold]Methods[/bold][br]The New Zealand neonatal TSH-based screening programme has prospectively identified potential cases of CHT using the same assay and screening thresholds since inception. Thyroid scintiscans were routinely recommended. We retrospectively identified all permanent CHT from 1993-2010 and calculated the ethnic specific incidence rates.[br][bold]Results[/bold][br]Over 18-years there were 330 new cases of CHT out of 1 053 457 live births in New Zealand (3.1/10 000 or 1:3192). Of the 86% who had a scintiscan, 67% had Thyroid Dysgenesis (female:male 5.0:1) and 33% Dyshormonogenesis (0.9:1). The overall incidence of CHT rose from 2.6/10 000 live births to 3.6/10 000 (p=0.01). There was increased incidence of dyshormonogenesis (p=0.02) but not dysgenesis, mediated by a two fold increase in Asian births, combined with a higher rate of dyshormonogenesis in Asians than NZ Europeans (Odds Ratio 3.3, p[lt]0.001). There was no change in the ethnic specific incidences of CHT in New Zealand over 18 years.[br][bold]Conclusions[/bold][br]These findings are reassuring that the overall increase in CHT incidence is due to changing demographics and there has not been an environmental influence on CHT rates in New Zealand.[br][br]Sources of Research Support: Ben Albert[apos]s position is supported by the Sir Graeme and Lady Douglas Fellowship.[br][br]Nothing to Disclose: BBA, CJ, DW, WSC, AJG, JC, KB, JGD, PLH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 788 190 1418 SUN-390 PO39-01 Sunday 1214 2012


1212 ENDO12L_SUN-391 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Study of Prevalence of Goiter in Oaxaca (Mexico): Analysis of Etiologic Factors Estanislao Ramirez-Vargas, Jaydi Nora Cruz-Fernandez, Rosalino Vasquez-Cruz Benito Ju[aacute]rez Autonomous University of Oaxaca (UABJO), Oaxaca, Mexico; Mexican Social Security Institute, Oaxaca, Mexico [bold][italic]Objectives:[/italic] [/bold]This study was planned to investigate goiter prevalence in the Valley of Oaxaca (Mexico), to analyze the etiologic factors and to study the correlation between goiter prevalence and the etiologic factors studied.[br][bold][italic]Methodology:[/italic][/bold] 1400 subjects (717 men and 683 women) of 10 to 18 years old were studied and engaged in clinical and laboratory tests. Physical examinations were: weight, size, BMI, palpation of neck for diagnosis of goiter by two experts[apos] endocrinologist; the goiter was classified according to the criteria of the WHO. Biological examinations were: Collect salt of various origins (gem, sailor, iodized) and collect water (taps and mineral water). Collect urines for determination of urinary iodine concentration ([mu]g iodine/mg of creatinine). Two recall of food consumption of 24 hours was administered with questionnaires.[br][bold][italic]Results: [/italic][/bold]The goiter prevalence in the population studied of Oaxaca was of 9.6% and the value of urinary iodine concentration was 79.8 [micro]g I/mg of creatinine (S.D. = 10.89). We found an important correlation between the prevalence of the goiter and the family history of goiter (O.R. = 2.4) and tobacco (O.R. = 2.2).[br][bold][italic]Discussion and Conclusions:[/italic][/bold] The goiter prevalence in the population studied of Oaxaca was 9.6%, near of the top threshold of endemia (according to I.C.C.I.D.D.). The value of the found average of urinary iodine was 79.8 [micro]g I/mg of creatinine (the recommendation of WHO is 150-200 [micro]g per day). We found an important correlation between the prevalence of the goiter and the family history of goiter and Tobacco addiction. No correlation was found between the presence of goiter and type of consumed salt, drunk water, the caloric consumption daily and the BMI.[br][br]Sources of Research Support: This study was supported by the Benito Ju[aacute]rez Autonomous University of Oaxaca (UABJO).[br][br]Nothing to Disclose: ER-V, JNC-F, RV-C 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2289 190 1419 SUN-391 PO39-01 Sunday 1215 2012


1213 ENDO12L_SUN-392 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Clinical Effects of Fixed Drug Holiday Compared to Daily Thyroxine Dosing in Primary Hypothyroid Patients Elliyyin Katiman, Karen Choong, Rokiah Pendek, Siew Pheng Chan University of Malaya, Kuala Lumpur, Malaysia [bold]Background: [/bold]Primary hypothyroidism is common and the current treatment recommendation is to give lifelong daily Levothyroxine (LT4) replacement. Noncompliance to daily LT4 replacement is the most common cause of inability to achieve euthyroid status. Limitation in availability of LT4 tablet strength (100 ug, 50 ug and 25 ug only) often complicates daily dosing logistics. We recommend an alternative regimen of LT4 replacement in the management of primary hypothyroid patients.[br][bold]Objectives: [/bold]To compare the effects of daily dosing versus drug holiday LT4 dosing on thyroid hormone, blood pressure, heart rate, weight, lipid profiles and quality of life by means of a health survey questionnaire.[br][bold]Methodology: [/bold]This is a 16-week randomized crossover control trial conducted in an academic research hospital involving 51 patients with stable primary hypothyroidism.The study was approved by the Institutional Medical Ethics Review Committee. Informed consent was obtained from all patients. The patients were randomized by block randomization to either daily dosing (DD) versus a fixed drug holiday (HD) LT4 dosing (five days a week LT4 with a 48 hour drug holiday over the weekend) over an 8 week period. After 8 weeks, the patients were switched to the opposite regimen for another 8 weeks of the study. The total cumulative dose of LT4 per week was the same during both treatment regimens. The thyroid profile, blood pressure, heart rate, weight, lipid profile and quality of life of patients were assessed at the beginning and at the end of each 8-week period of the study.[br][bold]Results: [/bold]Compared with daily dosing treatment, fixed drug holiday regimen showed no significant difference in the subjects thyroid profile and other parameters assessed. 28 patients (54.9%) preferred the HD regimen versus 19 patients (37.7%) that preferred the DD regimen. 4 patients (7.8%) had no special preferrence to either dosing regimen.[br][bold]Conclusion: [/bold]Fixed drug holiday LT4 dosing regimen was well tolerated and produced similar results in terms of thyroid hormone profile, blood pressure, heart rate, weight, lipid profiles and quality of life. Therefore, if supervised HD can be used as an alternative regimen which can improve patients[apos] ease of dosing especially in those with high noncompliance rate. More patients preferred HD regimen to the DD regimen.[br][br]Nothing to Disclose: EK, KC, RP, SPC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1047 190 1420 SUN-392 PO39-01 Sunday 1216 2012


1214 ENDO12L_SUN-393 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) A Pharmaco-Equivalent Replacement of Liothyronine for Levothyroxine Results in Similar Response to Escalating Doses of TRH Stimulation Test Sahzene Yavuz, Joyce D Linderman, Sheila Smith, Xiongce Zhao, Frank Pucino, Francesco S Celi NIDDK-National Institutes of Health, Bethesda, MD; NIDDK-National Institutes of Health, Bethesda, MD Clinical indications for TRH stimulation test are limited to the characterization of resistance to thyroid hormone, TSH-secreting adenoma, and central hypothyroidism. This test is also an investigational tool for the evaluation of the thyrotroph response to replacement therapy. We have previously demonstrated that a 1:3 mcg/mcg liothyronine (L-T3) for levothyroxine (L-T4) substitution results in equivalent TSH responses to a standard 200 mcg TRH injection. In this study we characterize the response to escalating doses of TRH in hypothyroid patients treated with equivalent doses of L-T3 or L-T4 to investigate subtle changes in the pituitary response to the two regimens. Patients were treated with L-T3 or L-T4 on a t.i.d. regimen to a target TSH of 0.5-1.5 mIU/l. Each subject underwent TRH stimulation tests with escalating TRH doses (5, 15 and 200 mcg) on both treatments. TRH was administered as an intravenous bolus, and the catheter was flushed with saline. Blood samples were drawn at -15, 0, 5, 10, 15, 20, 30, 60 minutes for measurement of TSH. Data were analyzed using repeated measures ANOVA and mixed-model analysis, and presented as delta from baseline (average -15[apos] and 0[apos]).[br]Thirteen hypothyroid patients (12 female) aged 51.2[plusmn]8.29 underwent escalating dose TRH stimulation testing. No significant differences were observed in peak TSH response 5 mcg: L-T3 6.05[plusmn]3.84 vs. L-T4 6.94[plusmn]5.59; 15 mcg: L-T3 6.34[plusmn]3.39 vs. L-T4 4.79[plusmn]4.61; and 200 mcg L-T3 6.8[plusmn]2.67 vs. L-T4 8.05[plusmn]7.01 mIU/l (p=n.s). Similarly AUC 5 mcg: L-T3 306.97[plusmn]188.14 vs. L-T4 352.08[plusmn]287.78; 15 mcg: L-T3 319[plusmn]172.15 vs. L-T4 241.48[plusmn]218.67; and 200 mcg L-T3 335.54[plusmn]130.35vs. L-T4 360.26[plusmn]341.66 (p =n.s.). L-T4 treatment resulted in a slightly but significantly shorter time to peak TSH response. 5 mcg L-T3 21.15[plusmn]6.50 vs. L-T4 19.61[plusmn]5.57; 15 mcg L-T3 21.53[plusmn]5.15 vs. L-T4 18.92[plusmn]4.46; and 200 L-T3 mcg 27.69[plusmn]4.38, vs. L-T4 24.23[plusmn]7.02 min, (P=0.02). Compared to the 5 mcg TRH dose, the 200 mcg dose resulted in small delay in TSH peak on both treatments L-T4 19.6 vs. 24.23 min and L-T3 21.15 vs. 27.69 min (p[lt]0.0001).[br]In conclusion our data confirm that substitution of L-T3 for L-T4 on a 1:3 mcg/mcg ratio using a t.i.d. administration regimen achieves similar degree of pituitary euthyroidism as compared to the standard L-T4 replacement. Furthermore, the data suggest that lower doses of TRH might be used to assess the thyrotroph response, particularly when investigating partial pituitary insufficiency states.[br][br]Sources of Research Support: This work was supported by the Intramural Research Program of the NIDDK-NIH. ClinicalTrials.gov identifier: NCT00106119.[br][br]Nothing to Disclose: SY, JDL, SS, XZ, FP, FSC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1004 190 1421 SUN-393 PO39-01 Sunday 1217 2012


1215 ENDO12L_SUN-395 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Should TSH Upper Reference Limit of Thyroid Function Reflex Tests Algorithms Be Reviewed? Alvaro Gonzalez, Estibaliz Alegre, Camen Perez-Ciriza, Ana Maria Chacon, Juan Carlos Galofre Cl[iacute]nica Universidad de Navarra, University of Navarra, Pamplona, Spain; Cl[iacute]nica Universidad de Navarra, University of Navarra, Pamplona, Spain In order to improve cost-effectiveness with no negative effect on diagnosis sensitivity, many laboratories perform reflex tests to study thyroid function. These tests are based on previously established algorithms which may study circulating thyroid hormone levels (free T4 and/or T3) depending on blood TSH level initial result. A crucial step in the design of these algorithms is to establish which serum TSH level cut-off will draw out additional T3 and free T4 analysis. Usually this cut-off is set up following both ends of TSH laboratory reference range.[br]Currently most laboratories consider that serum TSH upper reference range should be established around 5 mIU/L. However, some experts have recommended decreasing this upper limit to 2.5-3.0 mIU/L. Obviously, any fall in the superior TSH reference cut-off value would proportionately increase the number of thyroid hormone assays performed using these algorithms and will accordingly increase health care expenses.[br]We have retrospectively reviewed 25,453 thyroid function reflex test algorithms results carried out at our laboratory during a three-year period. The profile of rules in our algorithm was:[br]1) TSH level within the reference range (0.37-4.7 mIU/L): No further analyses were performed.[br]2) TSH level [lt]0.37 mIU/L: free T4 (RR: 9-24 pmol/L) and T3 (RR: 1,1-2,67 nmol/L) were investigated.[br]3) TSH level [gt]4.7 mIU/L: free T4 was measured.[br]We found 958 patients (3.76%) with serum TSH levels above 4.7 mIU/L during the study period, in whom 846 (88.3%) had normal free T4 in further analysis. Alternatively, if the upper TSH cut-off level had been established in 3.0 mIU/L, the percentage of additional free T4 tests would have increased by 263%. Likewise, an upper TSH cut-off level of 2.5 mIU/L would rise up to 450% the percentage of further free T4 analysis. Additionally we also observed that all subjects with TSH levels between 4.7 and 5.07 mIU/L (n=110) showed free T4 levels within the reference limit.[br]Considering the log-linear relationship between serum TSH and free T4 levels, low-ering the TSH cut-off in the diagnostic algorithms would considerably increase the number of additional free T4 tests, enhancing proportionally the expenses without any improvement in the algorithm sensitivity. Hence, we conclude that reflex tests to study thyroid function based on an initial serum TSH value need to maintain an upper cut-off level of 5 mIU/L.[br][br]Nothing to Disclose: AG, EA, CP-C, AMC, JCG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 213 190 1422 SUN-395 PO39-01 Sunday 1219 2012


1216 ENDO12L_SUN-396 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Changes in Thyroid Function in a HIV-1 Infected Population on Combined Antiretroviral Therapy Paula Freitas, Rita Bettencourt-Silva, Maria Joao Matos, Ana Cristina Santos, Sandra Xerinda, Antonio Sarmento, Jose Luis Medina, Davide Carvalho Centro Hospitalar S[atilde]o Jo[atilde]o, Faculty of Medicine, University of Porto, Porto, Portugal; Faculty of Medicine of Porto, Porto, Portugal; Centro Hospitalar S[atilde]o Jo[atilde]o, Faculty of Medicine of Porto, Porto, Portugal [bold]Introduction/aims:[/bold] Changes in thyroid function are common in HIV-infected patients. The prevalence of usually asymptomatic conditions (such as subclinical hypothyroidism) is increased and Graves[apos] disease may occur during immune reconstitution associated with combined antiretroviral therapy (cART). The aim of this study is to evaluate the presence of thyroid disease in HIV-1 infected patients on cART in a HIV Cardiometabolic Endocrine Out-patient Clinic. [bold]Patients and methods:[/bold] Cross-sectional study of 377 HIV-1 infected patients on cART. [bold]Results:[/bold] Three hundred and thirty-eight patients (89.7%) were euthyroid. Thyroid function changes were found in 10.3% (39/377): 1.3% (5/377) Graves[apos]s disease; 1.1% (4/377) Hashimoto[apos]s thyroiditis; 0.80% (3/377) papillary thyroid carcinoma; 0.5% (2/277) toxic multinodular goiter; 2.1% (8/377) subclinical hypothyroidism; 0.26% (1/377) postsurgical hypothyroidism; 0.53% (2/377) subclinical hyperthyroidism; 1.6% (6/377) low FT4 level and 2.1% (8/377) nonthyroidal illness syndrome. [bold]Conclusions:[/bold] The prevalence of thyroid function changes in this population was low when compared with other studies in HIV patients on cART (dysfunction prevalence of up to 35%), approaching the estimated prevalence in general population. The most common abnormalities were subclinical hypothyroidism and Graves[apos] disease.[br][br]Nothing to Disclose: PF, RB-S, MJM, ACS, SX, AS, JLM, DC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 114 190 1423 SUN-396 PO39-01 Sunday 1220 2012


1217 ENDO12L_SUN-397 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Thyroxine Treatment of Patients with Gastric Disorders: Preliminary Results with Softgel Capsules Maria Giulia Santaguida, Camilla Virili, Susanna Carlotta Del Duca, Lucilla Gargano, Marco Centanni [quot]Sapienza[quot] University of Rome, Rome, Italy; AUSL Latina, Latina, Italy The gastric acid producing machinery may be partially or totally destroyed in patients with atrophic body gastritis, blocked in those treated with proton pump inhibitors (PPI) and/or counteracted by NH3 production in those with Helicobacter pylori infection. An increased need for thyroxine has been described in some gastrointestinal disorders (H.pylori infection, gastric atrophy, celiac disease), even in their occult or symptomless forms (1-3). A transitory or stable impairment of gastric acidity, by increasing pH value, seems to interfere with thyroxine availability, highlighting a novel role for the stomach in the following intestinal T4 absorption. Distinctive dissolution profiles of different T4 preparations (tablets and/or soft gel capsules) have been described and soft gel T4 capsules performed better than tablets in alkaline pH. In patients with gastric disorders, where larger doses of thyroxine tablets are required, soft gel capsules of T4 may help to reach the therapeutic target and this represented the aim of our study. To this end, we have enrolled patients in therapeutic thyroid homeostasis, presenting with impaired gastric acid secretion and long-lasting T4 treatment ([gt]5 years) with the same brand of tablets. All these patients had been advised and agreed to take oral thyroxine under fasting conditions, waiting at least one hour before eating. Patients bearing additional conditions interfering with thyroxine treatment (e.g. drugs, intestinal disorders, pregnancy etc.) were excluded from the study. A total of 30 patients (28F/2M; median age=51 years; median T4 dose=2.05 [mu]g/kg/day) met these criteria and switched from the usual tablets treatment to the soft gel T4 capsules at a lower dose of thyroxine (median T4 dose=1.77 [mu]g/kg/day; p=0.0082). Thyroid function and TSH were measured after 3,6,12 and 18 months from the treatment switch. Most of patients (18/30; 60%) showed a TSH increase after 3 months of treatment, with no change in FT4 levels. However, after 6 months, TSH returned to the baseline values in about 2/3 of patients despite the reduced dose of T4. Thyroid hormones were not changed during the study (p=ns). These preliminary findings strongly suggest that treatment with soft gel T4 capsules may reach the therapeutic goal at a lower dose than a T4 tablet preparation in patients with impaired gastric acid secretion.[br][br](1)Centanni M et al.,New Engl J Med 2006; 354(17):1789-1795. (2)Centanni M et al. hotthyroidology.com 2007, n.169:1-4. (3)Virili C et al. J Clin Endocrinol Metab 2012[Epub ahead of print].[br][br]Nothing to Disclose: MGS, CV, SCDD, LG, MC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 458 190 1424 SUN-397 PO39-01 Sunday 1221 2012


1218 ENDO12L_SUN-398 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Effect of Gastric pH on the Bioavailability of Levothyroxine (LT4) in Soft Gel Capsules and in Tablets: Results from a Pharmacokinetic (PK) Study in Healthy Volunteers Salvatore Benvenga, Murray Ducharme University of Messina, Policlinico G Martino, Messina, Italy; University de Montreal, Montreal, Canada In vitro studies by Pabla et al(1) showed that dissolution of LT4 at various pH values is much more consistent when LT4 is contained in a soft gelatin capsule (Tirosint[reg] capsule, IBSA, Switzerland) rather than in a conventional brand tablet or a generic tablet. These dissolution data allowed to predict better performance of capsules over tablets upon real-life challenges on LT4 absorption, caused by natural or drug-induced alkalinization of the physiologically acid gastric pH (e.g., atrophic gastritis, proton pump inhibitors [PPI]).[br]To investigate the effect of gastric pH on the bioavailability of LT4 in the same soft gel capsules and in tablets (Synthroid[reg], Abbott Laboratories, USA), 16 healthy male volunteers were administered 600 mcg of LT4 with 240 mL water after a 10-h overnight fast, in a single dose, randomized, 2-way cross-over study. Before LT4 administration, subjects were infused i.v. 80 mg of esomeprazole (Nexium[reg], AstraZeneca AG, Switzerland) in 30 min and gastric pH was monitored by means of a nasogastric tube pre-dose and until 5 h post-dose. The acute treatment was aimed to avoid interferences and standardise conditions. Blood samples were obtained prior to and at several times up to 24 h after dosing to assay serum total T4. The main PK parameters were calculated after baseline subtraction and ln-transformed data were compared for capsule and tablet using ANOVA for a cross-over design.[br]In a condition of altered pH (mean pH over 2 h after dosing ranged 4.2-8.5), mean area under the curve (AUC) was 19% higher (90% CI: 99.5-141%) and mean peak concentration (Cmax) was 23% higher (90% CI: 103-138%) with the capsule than with the tablet. Mean Tmax was 23% lower with the capsule. At all time points from 1 h through 24 h, mean serum T4 values were greater for the capsule, and perfect parallelism between the two concentration-time curves existed from 4 h on.[br]The bioequivalence between Tirosint[reg] and Synthroid[reg] in normal conditions had already been demonstrated in a previous study(2). In the present study, the capsule showed superiority over the tablet in terms of LT4 absorption in healthy volunteers with gastric pH increased by administration of PPI. This might solve the problem of L-T4 tablet malabsorption caused by simultaneous therapy with PPI. These differences between the capsule and the tablet (in favor of the capsule) add to the superiority demonstrated in patients with coffee-related L-T4 malabsorption(3).[br][br](1) Pabla D et al, Eur J Pharm Biopharm. 2009;72(1):105-10. (2) Colucci P et al, Ther Drug Monit. 2011;33(3):355-61. (3) Benvenga S et al. ATA, 2011; abstract 196.[br][br]Nothing to Disclose: SB, MD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 800 190 1425 SUN-398 PO39-01 Sunday 1222 2012


1219 ENDO12L_SUN-399 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Pathophysiology-Based Review of Food and Drug Interactions with Levothyroxine Therapy Fatemeh D Mojdami, Alyse Goldberg, Rommel G Tirona, Stan H Van Uum University of Western Ontario, London, Canada Introduction [amp] Aim: Levothyroxine is the fourth most prescribed drug in the United States in 2010, and is used in primary therapy for hypothyroidism and differentiated thyroid cancer. Pharmacologic interference of levothyroxine action by drugs and food substances is well-established and remains a challenge in clinical practice, particularly in the elderly population where polypharmacy is common. Most reviews of these interactions are not based on underlying pathophysiological mechanisms, and do not review the strength of evidence for the proposed interactions. We reviewed the literature to classify proposed mechanisms for interactions and assessed the accompanying evidence.[br]Methods: We searched the literature for drugs, foods and supplements affecting levothyroxine pharmacokinetics using available scientific databases including MEDLINE, EMBASE, Scopus and Web of Science. A total of 69 citations were identified between 1980 to June 2011. These articles were reviewed for the agent of interference, the proposed mechanism, the thyroid status of subjects at baseline, the duration of the proposed interaction and the level of supporting evidence.[br]Results: Several agents interfering with levothyroxine pharmacokinetics have been identified. The level of evidence for these interactions varies significantly with case reports being common and prospective studies being rarer. Multiple mechanisms have been proposed with the most common being interference with levothyroxine absorption in the gastrointestinal tract, including the effect of calcium and iron. Other proposed mechanisms, with variable degree of experimental evidence, include interference with protein-binding, efflux/influx transporters, deiodination, hepatic metabolism, excretion, nuclear protein binding and effects on the hypothalamus and pituitary gland.[br]Discussion: We have catalogued several proposed mechanisms for interactions between drug and food products with levothyroxine therapy, and graded the strength of the evidence for each agent. The most common mechanism is interference with absorption in the gut. Clinically relevant interference is more likely in those with a baseline diagnosis of hypothyroidism. We established a framework for a pathophysiological approach to assess these interactions, and highlight areas of knowledge deficiency that require further investigation.[br][br]Nothing to Disclose: FDM, AG, RGT, SHVU 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1347 190 1426 SUN-399 PO39-01 Sunday 1223 2012


1220 ENDO12L_SUN-400 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Rapid Progression to Overt Hypothyroidism with Very High Levothyroxine Requirements Following Initiation of Antiepileptic Drugs Marc Laufgraben, Zeina Maani Cooper Medical School of Rowan University, Camden, NJ; Alpert Medical School of Brown University, East Providence, RI [bold]Background: [/bold]Antiepileptic drugs (AEDs) are known to affect thyroid hormone metabolism but generally cause only mild abnormalities. We report the case of a woman with a history of subclinical hypothyroidism who had rapid progression to overt hypothyroidism with very high levothyroxine (LT4) requirements following initiation of AEDs.[br][bold]Clinical case: [/bold]A 46 year-old woman was admitted with septic shock due to aspiration pneumonia. Four years prior to admission, thyroid function tests (TFTs) demonstrated a slightly elevated TSH of 6.16 uIU/ML (nl 0.35-5.5) with Free T4 0.81 NG/DL (nl 0.8-1.8). During her hospitalization, the patient developed seizures and was started on levetiracetam, later requiring the addition of phenobarbital and phenytoin. Two months after admission, her TSH was found to be slightly elevated at 8.72 with a low free T4 of 0.65. Her weight was 40 kilograms, and she was started on LT4 12.5 mcg PO daily. TFTs were rechecked two months later: TSH was 85.8 with free T4 0.41. LT4 was changed to IV, then increased to 25 mcg IV daily and later to 50 mcg IV daily over the next six weeks with minimal change in TFTs. Endocrine consultants diagnosed increased thyroid hormone clearance due to AEDs. LT4 dose was increased further to 100 mcg IV, with improvement in TSH to 21.3, and free T4 to 0.72. TFTs normalized with LT4 125 mcg IV daily. Once she was safely tolerating oral intake, she was changed to 200 mcg PO daily and discharged on that dose.[br][bold]Conclusion: [/bold]AEDs--particularly older drugs such as phenytoin, phenobarbital, carbamazepine and primidone--may increase thyroid hormone clearance by induction of cytochrome P450; induction of UDP glucuronosyltransferases may also play a role. Patients with normal thyroid function do not typically experience problems on AEDs. However, our patient likely had limited thyroid reserve based on a history of subclinical hypothyroidism, and could not adapt to AED-induced increases in thyroid hormone clearance, resulting in overt hypothyroidism with very high thyroid hormone requirements for her weight. Patients with limited thyroid reserve (as suggested by a history of subclinical thyroid disease or antithyroid antibodies) may experience progression to overt hypothyroidism on AEDs, and monitoring of TFTs is warranted, particularly in the initial months of therapy. Likewise, patients on thyroid hormone replacement should have TFTs monitored whenever AEDs are initiated, titrated or discontinued.[br][br]Nothing to Disclose: ML, ZM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 41 190 1427 SUN-400 PO39-01 Sunday 1224 2012


1221 ENDO12L_SUN-401 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Prevalence, Relative Risk and Clinical Associations of Hypothyroidism in Patients with Type 2 Diabetes Juan J Diez, Pedro Iglesias Hospital Ram[oacute]n y Cajal, Madrid, Spain Background. The prevalence of thyroid dysfunction has been found to be elevated in patients with both type 1 and type 2 diabetes (T2D). However, most of the surveys have not reported a link between thyroid hypofunction and diabetes-related clinical and analytical parameters.[br]Aims. To determine the relative risk of hypothyroidism in patients with T2D, and evaluate predictive factors for hypothyroidism in our diabetic population.[br]Patients and methods. 1112 patients (58% females, mean age 66.7 yr, median duration of diabetes 10 yr) were evaluated. There were 179 patients with previous history of thyroid dysfunction. A sample of 911 non-diabetic subjects without known thyroid dysfunction was studied as control group.[br]Results. Prevalence of overall hypothyroidism in the studied diabetic population was 18.3% (8.5% subclinical and 9.9% overt). Newly diagnosed hypothyroidism was found in 7.3% of our patients (0.4% overt and 6.9% subclinical). When compared diabetic patients without previously known thyroid dysfunction with control subjects, the odds ratio (OR) for hypothyroidism in diabetics was 2.68 (1.69-4.25). This OR was significant in patients over age 65 yr (3.96 [1.92-8.17]), as well as in both males (4.27 [1.45-12.56]) and females (2.56 [1.52-4.31]), obese (2.56 [1.36-4.81]) and non-obese (3.11 [1.56-6.20]) patients and in patients with (4.17 [1.72-10.11]) and without (2.93 [1.49-5.73]) thyroid autoantibodies. Logistic regression analysis showed that newly diagnosed hypothyroidism was significantly and directly related not only with thyroid autoimmunity (19.15 [8.73-42.01]), but also with the presence of macroangiopathy (2.85 [1.51-5.35]) and metformin treatment (2.51 [1.28-4.92]). No other significant relationships between hypothyroidism and diabetes-related clinical parameters were found.[br]Conclusion. Our results are in favor of the screening for hypothyroidism in patients with T2D older than 65 yr, regardless of gender and weight status. The presence of thyroid autoantibodies, diabetic macroangiopathy and metformin treatment seem to be predictive factors for the presence of undetected hypothyroidism in diabetic patients.[br][br]Nothing to Disclose: JJD, PI 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 206 190 1428 SUN-401 PO39-01 Sunday 1225 2012


1222 ENDO12L_SUN-402 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Effect of L-Thyroxin Therapy on Thyroid Volume and Carotid Artery Intima-Media Thickness in Patients with Subclinical Hypothyroidism Ilknur Ozturk Unsal, Oya Topaloglu, Nujen Bozkurt Colak, Basak Karbek, Askin Gungunes, Muyesser Sayki Arslan, Esra Tutal Akkaymak, Bekir Ucan, Taner Demirci, Melia Karakose, Mustafa Caliskan, Erman Cakal, Tuncay Delibasi Diskapi Yildirim Beyazit Teaching and Research Hospital, Ankara, Turkey [bold]Backround:[/bold]Subclinical hypothyroidism(SCH) is mild-to-moderate thyroid insufficiency that is characterized by thyrotropin(TSH) level higher than the upper limit despite normal serum free thyroxin (fT4) level.The prevalence of SCH is 4-10%.It is known that manifest hypothyroidism is a risk factor for cardiovascular diseases(CVD) and atherosclerosis.Likewise, SCH as well might be a risk factor for cardiovascular diseases.Some placebo-controlled studies showed beneficial effect of L-thyroxin(L-T4) replacement on the risk for early atherosclerosis and CVD in the patients with SCH.Measurement of the carotid artery intima-media thickness(C-IMT)via B-mode ultrasonography provides easy evaluation of atherosclerosis.[br][bold]Methods:[/bold]Fifty-six patients presented to our clinic with subclinical hypothyroidism were included in the study.Forty-six healthy euthyroid subjects were included as the control group.Patients with fT4[gt]0.61ng/dl and TSH[gt]4.2 uIU/ml were considered as SCH.Carotid artery intima-media thickness (CIMT) was measured via B-mode ultrasonography and thyroid volume was calculated. L-T4replacement was commenced at a dose of 25-50 mcg/day.CIMT and thyroid volumes of the patients were reevaluated six months after they became euthyroid.[br][bold]Results:[/bold]Of the patients in the SCH group, 51(91.1%) were female and 5(8.9%) were male, whereas the control group comprised 25(54.3%) females and 21(45.7%) males.A statistically significant difference(p[lt]0.05) was found between the CIMT values before and after the L-thyroxin therapy in the SCH group.Pretreatment CIMT values were significantly higher than the post-treatment CIMT values.There was significant difference also between the pre-treatment and post-treatment thyroid volumes in the SCH group(p[lt]0.05); pretreatment thyroid volumes were significantly higher than the post-treatment thyroid volumes.No significant difference was determined between the SCH and the control group in terms of pretreatment CIMT values (p[gt]0.05).When the SCH group was divided into two as TSH [lt]7 uIU/ml (n=39) and TSH [ge]7 uIU/ml (n=17), no significant difference was found between the groups in terms of CIMT values and thyroid volumes(p[gt]0.05).[br][bold]Conclusion:[/bold]The present study showed the reduction in CIMT with L-T4 therapy in the patients with subclinical hypothyroidism.Therefore, thyroid hormone replacement might help to slow down or prevent atherosclerosis in the subclinical hypothyroidism as well.[br][br]Nothing to Disclose: IOU, OT, NBC, BK, AG, MSA, ETA, BU, TD, MK, MC, EC, TD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1430 190 1429 SUN-402 PO39-01 Sunday 1226 2012


1223 ENDO12L_SUN-403 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Preoperative Thyroid Dysfunction Predicts 30-Day Postoperative Complications in Elderly Hip Fracture Patients Treated Surgically Xi Wern Ling, Tet Sen Howe, Joyce Suang Bee Koh, Merng Koon Wong, Alvin Choong Meng Ng Singapore General Hospital, Singapore, Singapore; Singapore General Hospital, Singapore, Singapore [bold]Objectives:[/bold] To investigate the relationship between thyroid function and short-term outcomes in elderly patients with hip fracture who are managed surgically.[br][bold]Design:[/bold] Retrospective observational cohort study.[br][bold]Setting:[/bold] Orthopaedic ward in Singapore General Hospital.[br][bold]Participants:[/bold] Patients admitted from July 2009 to June 2010 with low-energy hip fracture. Excluded were patients aged [lt]60 years, those with pathological or periprosthetic fracture and those whose fractures were treated nonoperatively.[br][bold]Methods:[/bold] Serum TSH and free T4 as well as routine blood chemistry was measured preoperatively as part of a Coordinated Clinical Pathway for hip fracture patients. Patients were classified as having biochemically overt or subclinical hyperthyroidism or hypothyroidism, normal thyroid function or sick euthyroid state based on serum TSH and free T4 values. Outcome data was collected from hospital records and discharge summaries. Biochemical thyroid dysfunction was not systematically treated.[br][bold]Outcome measure:[/bold] Postoperative complications at 30 days.[br][bold]Results:[/bold] A total of 254 patients were analysed; 64 (25.2%) were male and mean age was 77.8 years. There were 128 euthyroid (50.4%), 68 (26.8%) sick euthyroid, 13 (5.1%) overtly hyperthyroid, 20 (7.9%) subclinically hyperthyroid, 4 (1.6%) overtly hypothyroid and 21 (8.3%) subclinically hypothyroid patients. Complications developed in 96 patients (38%) within 30 days of surgery. The most common complication was urinary tract infection (32 patients, 12.6%), followed by cardiac events (21 patients, 8.3%) and delirium (14 patients, 5.5%). Only patients with overt hyperthyroidism were found to be at increased risk of complications in multivariate analysis (odds ratio (OR) = 4.4, 95% confidence interval (CI) = 1.3-15.2). Complications in this group were similar to those in the overall cohort. Thyroid function was not predictive of length of hospital stay, 30-day mortality or readmissions.[br][bold]Conclusion:[/bold] Older patients frequently develop complications following surgery for hip fractures. This risk appears to be increased by preoperative biochemically overt hyperthyroidism. Further study is warranted to confirm this finding and to determine if treatment of the hyperthyroid state preoperatively improves outcomes.[br][br]Nothing to Disclose: XWL, TSH, JSBK, MKW, ACMN 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 736 190 1430 SUN-403 PO39-01 Sunday 1227 2012


1224 ENDO12L_SUN-404 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Leptin and Serum Thyrotropin Concentrations, Thyroid Autoimmunity and Smoking Status, in a Representative Sample of Iodine-Sufficient, Euthyroid Mediterranean Population with Different Body Mass Index Anna Lucas-Martin, Maria Luisa Granada, Izaskun Olaizola, Conxa Castell, Maria Teresa Julian, Silvia Pellitero, Manuel Puig-Domingo Germans Trias i Pujol Hospital, Badalona, Spain; Autonomous University of Barcelona, Badalona, Spain; Generalitat of Catalonia, Barcelona, Spain; Germans Trias i Pujol Hospital, Badalona, Spain [bold]Introduction[/bold][br]A positive correlation between serum thyrotropin concentrations (TSH) and body mass index (BMI) has been demonstrated in different but not all studies in euthyroid subjects, or observed only if thyroid autoimmunity (TA) is present or in non-smokers individuals (NS). Leptin (Lp) could be the major link between BMI and TSH.[br][bold]Design[/bold][br]To analyze the relationship between TSH, free thyroxin (FT[sub]4[/sub]), Lp, TA (peroxidase and/or thyroglobulin antibodies) and smoking status in a representative sample of euthyroid, iodine-sufficient, non-hospitalized population of Catalonia, with different BMI. Data collected included smoking habit. Glycemia and insulinemia were measured and HOMA index, calculated.[br][bold]Results[/bold][br]894 adults (390 men) of 44.87[plusmn]15.03 years and BMI 26.19[plusmn]4.61 Kg/m[sup]2[/sup] (17.01[ndash]52.70) with normal TSH (0.33[ndash]3.96 mIU/L) and FT[sub]4[/sub] (0.87[ndash]1.90 ng/dL), and median urine iodine concentration 150.0 mg/L were studied.[br]Lp correlated directly with BMI in both sexes (p=0.000). There was no correlation between TSH and BMI. In men, TSH correlated directly with Lp (p=0.004) and in women, directly with Lp (p=0.002) and HOMA (p=0.031) and inversely, with FT[sub]4[/sub] (p=0.024). Only in smoker men, TSH correlated directly with Lp (p=0.010) and HOMA (p=0.024). In smoker women, TSH correlated directly with Lp (p=0.004) and in NS women, inversely with FT[sub]4[/sub] (p=0.047).[br]Multivariate analysis showed Lp ([beta]=0.1304, p=0.025) and age ([beta]=-0.0051, p=0.012) to be independent predictors of TSH variations in men and Lp, the presence of TA ([beta]=0.1168, p=0.042; [beta]=0.2591, p=0.001) and FT[sub]4[/sub], ([beta]=-0.1395, p=0.018) in women.[br][bold]Conclusions[/bold][br]Leptin is a predictor factor of TSH concentration variations, in euthyroid subjects of both sexes. Other predictor factors are different in men and women. The smoking status can influence the relationship between TSH and some predictor factors. These data should be taken into account before drawing conclusions about the parameters that influence TSH concentrations in euthyroid people with different BMI.[br][br]Nothing to Disclose: AL-M, MLG, IO, CC, MTJ, SP, MP-D 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 388 190 1431 SUN-404 PO39-01 Sunday 1228 2012


1225 ENDO12L_SUN-405 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Insulin Resistance and Thyroid Function Status in Inner City Obese Minority Youths Jessica May Gold, Megan Kathleen Long, Leslie Lam, Morri Markowitz, Ping Zhou Children[apos]s Hospital at Montefiore, Bronx, NY; Albert Einstein College of Medicine, Bronx, NY; Children[apos]s Hospital at Montefiore, Bronx, NY [bold]Background:[/bold] Elevated TSH level with normal thyroid hormone levels and insulin resistance have been described independently in obese subjects.[br][bold]Objective[/bold]: To examine the relationship between thyroid function tests, insulin resistance, and their related metabolic manifestation in an obese minority pediatric population.[br][bold]Methods[/bold]: This is an IRB-approved retrospective study. Data is collected from the records of 298 obese, Hispanic or African American patients, ages 6 to 21.[br]Subjects with chronic medical conditions, such as with positive auto-antithyroid antibodies or on medications are excluded. Data collected includes TSH, free T4, fast insulin, glucose, lipid profile, and anthropometric measures. Calculated measures include BMI, BMI Z-score, and QUICKI (a measure of insulin sensitivity). QUICKI values of less than 0.34 indicate insulin resistance.[br]The statistical analyses performed in this study include Pearson correlations and comparison of means of groups stratified by TSH with a low TSH group (n=47, TSH [lt]1.0) and a high TSH group (n=16, TSH[gt] 4.0).[br][bold]Results[/bold]: TSH (1.92[plusmn]1.10, range 0.04-8.92 uU/mL), free T4 (1.28[plusmn]0.24, range 0.7-2.53 ng/dL). These indicate that our population is generally euthyroid with a mild degree of TSH variation. TSH is positively correlated with BMI Z-score (r=0.152, p=0.0089); Quick index is negatively correlated with BMI (r=-0.246, p=0.00002), BMI Z-score (r=-0.223, p=0.00007), and triglycerides (r=-0.25, p=0.0004). Interestingly, TSH is positively correlated with HDL (r=0.23, p=0.0009). There is no correlation between TSH and Quicki index.[br]The mean of BMI Z-score is higher and the mean QUICKI is lower in the high TSH group than in the low TSH group. They are 2.41 vs. 2.17 (p=0.0465) and 0.30 vs. 0.32(p=0.0466) respectively. However, after adjusting for BMI, the difference in the QUICKI becomes statistically insignificant, which suggests that the difference of insulin resistance between the two groups can be attributed to obesity.[br][bold]Conclusions[/bold]: Our results support previous studies that TSH and insulin sensitivity are independently associated with the degree of obesity. However, our results do not point to elevated TSH level ([ldquo]TSH resistance[rdquo]) as a mediator of insulin resistance or the latter as explaining [quot]TSH resistance[quot] in obese minority children. Our data also suggest that metabolic consequences of obesity, such as dyslipidemia, are from insulin resistance rather than [ldquo]TSH resistance[rdquo].[br][br]Fernandez-Real et al. Thyroid Function Is Intrinsically Linked to Insulin Sensitivity and Endothelium-Dependent Vasodilation in Healthy Euthyroid Subjects. J Clin Endorinol Metab, 2006; 91(9): 3337-3343. Ganie et al. Association of subclinical hypothyroidism and phenotype, insulin resistance, and lipid parameters in young women with polycystic ovary syndrome. Fertility and Sterility, 2011;95(6): 2039-2043. Mueller, A., Sch[ouml]fl, C., et al. Thyroid-stimulating hormone is associated with insulin resistance independently of body mass index and age in women with polycystic ovary syndrome. Human Reproduction, 2009;24(11):2924-2930. Nader et al. Relationships Between Thyroid Function and Lipid Status or Insulin Resistance in a Pediatric Population. Thyroid, 2010; 20(12):1334-1339. Roos, Annemieke, Bakker, Stephan J. L, et al. Thyroid Function Is Associated with Components of the Metabolic Syndrome in Euthyroid Subjects. J Clin Endocrinol Metab, February 2007, 92(2):491[ndash] 496. Shalitin, S., Yackobovitch-Gavan, M., et al. Prevalence of Thyroid Dysfunction in Obese Children and Adolescents before and after Weight Reduction and Its Relation to Other Metabolic Parameters. Horm Res 2009;71:155-161.[br][br]Nothing to Disclose: JMG, MKL, LL, MM, PZ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1094 190 1432 SUN-405 PO39-01 Sunday 1229 2012


1226 ENDO12L_SUN-406 POSTER SESSION: Hypothyroidism, Pregnancy, [amp] Pediatric Thyroid Disease (1:30 PM-3:30 PM) Evaluation of Thyroid Function Tests and Thyroid Ultrasound in Patients with Obstructive Sleep Apnea Syndrome Yavuz Yalcin, Dilek Berker, Turkan Mete, Oya Topaloglu, Bulent Ciftci, Selma Firat Guven, Yasemin Tutuncu, Ferhat Gokay, Serdar Guler Numune Education and Research Hospital, Ankara, Turkey; Atat[uuml]rk Chest Diseases and Thoracic Surgery Education and Research Hospital, Ankara, Turkey [bold]Objectives: [/bold]Obstructive sleep apnea syndrome (OSAS) is a disorder characterized by recurrent, parsial or complete episodes of apnea due to upper airway tract obstruction during sleeping period. There are some similarities between thyroid dysfunction such as hypothyroidism and OSAS in clinical features. Hypothyroidism is a predisposing factor for OSAS. Thus, in order to evaluate thyroid dysfunction, most studies have been focused on hypothyroidism in OSAS patients. This study was designed to determine prevalence of thyroid disease in OSAS patients based on thyroid function tests, thyroid antibodies, and thyroid ultrasound.[br][bold]Methods[/bold]: In this prospective study, 150 OSAS patients (50 patients with mild, 50 with moderate, 50 with severe OSAS) and 32 non-OSAS controls who were referred to the sleep disorders center for polysomnography test from September 2010 to November 2011 were included in the study. Serum thyroid- stimulating hormone (TSH), free- thyroxine (fT4), free-triiodothyronine (fT3) levels, anti-thyroid-peroxidase (anti-TPO), and anti- thyroglobulin (anti- TG) antibodies were measured in all patients within 4 weeks of the sleep study. Thyroid ultrasound was performed in endocrinology outpatient clinic with General Electric LOGIQ 3 EXPERT ultrasound machine with 11 mHz linear probe. The thyroid parenchyma echogenicity determined by ultrasonographically was classified as homogenous (normal or diffusely enlarged thyroid gland), heterogenous (irregularly increased echogenicity), nodular (solitary or multinodulated).[br][bold]Results[/bold]: We did not find statistically significant difference in prevalence of subclinical hypothyroidism, overt hypothyroidism and hyperthyroidism between OSAS patients and controls. Ultrasonographic prevalence of nodules in OSAS patients was 42% whereas in controls it was 43.7 %. There was no statistically significant difference in number of nodules determined by ultrasonographically between controls and patients (p.0.966). Thyroid autoantibody levels were similar in both groups. Also, we did not find any difference in thyroid parenchyma heterogenicity between groups (p.0.742).[br][bold]Conclusion:[/bold] The prevalence of subclinical hypothyroidism, overt hypothyroidism and hyperthyroidism was similar in OSAS patients and in controls. We consider that long-term follow-up of these patients will clarify if routine thyroid function tests screening is necessary or not.[br][br]Nothing to Disclose: YY, DB, TM, OT, BC, SFG, YT, FG, SG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1597 190 1433 SUN-406 PO39-01 Sunday 1230 2012


1227 ENDO12L_SUN-407 POSTER SESSION: Thyrotoxicosis, TSH Administration [amp] Thyroid Cell Biology (1:30 PM-3:30 PM) Novel Diagnostic Criteria and Clinico-Epidemiological Features of Thyroid Storm Based on a Japanese Nationwide Survey Takashi Akamizu, Tetsurou Satoh, Osamu Isozaki, Atsushi Suzuki, Shu Wakino, Tadao Iburi, Kumiko Tsuboi, Tsuyoshi Monden, Tsuyoshi Kouki, Naotetsu Kanamoto, Hajime Otani, Satoshi Teramukai, Masatomo Mori Wakayama Medical University, Wakayama, Japan; Gunma University Graduate School of Medicine, Maebashi, Japan; Tokyo Women[apos]s Medical University, Tokyo, Japan; Fujita Health University, Nagoya, Japan; Keio University, Tokyo, Japan; Tokyo Medical and Dental University Graduate School, Tokyo, Japan; Toho University School of Medicine, Tokyo, Japan; Dokkyo University School of Medicine, Tokyo, Japan; University of the Ryukyus, Okinawa, Japan; Kyoto University Graduate School of Medicine, Kyoto, Japan; Kansai Medical University, Hirakata, Japan; Kyoto University School of Medicine, Kyoto, Japan Thyrotoxic storm is a life-threatening condition requiring emergency treatment. The condition arises in thyrotoxic patients who manifest decompensation in multiple organs, often triggered by severe stress. Neither its epidemiological data nor diagnostic criteria have been fully established. We attempted to clarify clinico-epidemiological characteristics of thyroid storm patients using a nationwide survey and then formulate novel diagnostic criteria for thyroid storm. Further, we attempted to create guidelines for the management and treatment of thyroid storm, based upon the nationwide survey. We first developed tentative diagnostic criteria (the first edition) mainly based upon the literature. The first edition of the diagnostic criteria defined definite and suspected cases based on a prerequisite item (the presence of thyrotoxicosis) and combinations of typical clinical features (fever [[ge]38[deg]C], tachycardia [[ge]130 bpm], CNS [restlessness, delirium, mental aberration/psychosis, somnolence/lethargy, convulsion or coma, 14[ge] the GCS, 1[le] the Japan Coma Scale] and GI/hepatic [nausea, vomiting, diarrhea and jaundice] signs and symptoms, and CHF [pulmonary edema, moist rales over more than half the lung field, or cardiogenic shock, Class IV by the NYHA classification or Class III or higher by the Killip classification]). We then performed the nationwide survey on thyroid storm using these criteria, targeting all hospitals in Japan with eight-layered random extraction. The first nationwide survey revealed that the number of definite and suspected thyroid storm cases was estimated to be 1,283 [plusmn] 105 (95% confidence interval: 1,077 [minus] 1,489) per five years; approximately 260 per year (0.20 persons per 100,000 Japanese population per year) and 0.22 % of thyrotoxic patients. The second nationwide survey obtained detailed clinico-epidemiologic features of 282 definite and 74 suspected cases. The mortality rates of definite and suspected cases were 11.0% and 9.5%, respectively. Finally, based on the results of this survey, we propose a revision of the diagnostic criteria (the second edition). In conclusion, thyrotoxic storm is still a life-threatening disorder with over 10% mortality in Japan. We formulated novel diagnostic criteria for this disorder. We hope that they will contribute to quick and precise clinical decisions and treatment of this disorder, ultimately achieve better prognosis of thyroid storm.[br][br]Nothing to Disclose: TA, TS, OI, AS, SW, TI, KT, TM, TK, NK, HO, ST, MM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 121 191 1434 SUN-407 PO39-02 Sunday 1231 2012


1228 ENDO12L_SUN-408 POSTER SESSION: Thyrotoxicosis, TSH Administration [amp] Thyroid Cell Biology (1:30 PM-3:30 PM) Risk Factors for Treatment Failure, Indicators of Incipient Hypothyroidism and Assessment of Weight Gain Following Radioiodine Therapy for Graves Thyrotoxicosis Fraser W Gibb, Roger W Brown, Shareen Forbes, Alan Jaap, Alan W Patrick, Rebecca M Reynolds, Brian R Walker, Nicola N Zammitt, Mark W Strachan NHS Lothian, Edinburgh, UK BACKGROUND[br]Radioiodine ([sup]131[/sup]I) therapy for the management of Graves[apos] thyrotoxicosis is well tolerated but has been associated with excess weight gain and progression of dysthyroid eye disease, which may be attributable to late recognition of developing hypothyroidism. We sought to assess hypothyroidism following [sup]131[/sup]I administration and the extent to which following ATA guidelines mitigates late diagnosis. We also aimed to identify predictors of treatment failure, incipient hypothyroidism and weight gain.[br]METHODS[br]All patients in our unit receiving [sup]131[/sup]I for Graves[apos] disease between 2006 and 2010 (n=288, 77% female) were identified. Baseline parameters including TSH, free thyroxine (fT4), TSH receptor antibody levels, weight, age and exposure to thionamides were ascertained. In 80 patients, [sup]131[/sup]uptake scans were performed prior to therapy. 11mCi [sup]131[/sup]I was administered to all but 9 patients (who received 22mCi). The results of all post treatment thyroid function tests were recorded. At one year, weight gain was assessed and patients were classified as euthyroid, hypothyroid or hyperthyroid (including those who required further [sup]131[/sup]I).[br]RESULTS[br]One year after [sup]131[/sup]I 80.9% of patients had developed hypothyroidism (all patients receiving 22mCi), 13.5% remained thyrotoxic and 5.6% were euthyroid (no age or gender difference). 28.2% of hypothyroidism developed in the first 60 days after radioiodine and 54.5% of patients had TSH levels greater than 20mU/L at diagnosis. Treatment failure occurred in 21.8% of patients pre-treated with thionamides compared with 6.2% of those with no prior exposure (p[lt]0.001) although radioiodine uptake was higher in thionamide treated patients (p[lt]0.01), suggesting more severe disease. During follow up, development of a detectable TSH or fT4 levels less than 15 pmol/L beyond 4 weeks was never associated with recurrent thyrotoxicosis. At one year, mean weight gain was 4.4% in those in the lowest tertile of pre [sup]131[/sup]I fT4, but was 12.3% in the highest(p[lt]0.001).[br]DISCUSSION[br]Post [sup]131[/sup]I hypothyroidism often develops rapidly and, despite following ATA guidelines in the majority of patients, a large proportion became significantly hypothyroid before thyroxine replacement was initiated; this may account for some [sup]131[/sup]I related adverse outcomes. Following [sup]131[/sup]I, thyroid autonomy is uncommon in patients with low-normal fT4 levels or detectable TSH, so a strategy of pre-emptive thyroxine replacement, in this context, could be safe and potentially limit morbidity.[br][br]Nothing to Disclose: FWG, RWB, SF, AJ, AWP, RMR, BRW, NNZ, MWS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1079 191 1435 SUN-408 PO39-02 Sunday 1232 2012


1229 ENDO12L_SUN-409 POSTER SESSION: Thyrotoxicosis, TSH Administration [amp] Thyroid Cell Biology (1:30 PM-3:30 PM) Subclinical Thyroid Dysfunction Is Not Associated with Hip Fracture or Lower Bone Mineral Density in Older Adults Margaret C Garin, Alice M Arnold, Jennifer S Lee, John A Robbins, Anne R Cappola University of Pennsylvania, Philadelphia, PA; University of Washington, Seattle, WA; University of California Davis, Sacramento, CA; University of California Davis, Sacramento, CA [bold]Background:[/bold] Subclinical thyroid dysfunction is common in older individuals and has been linked to increased fracture risk. We assessed the risks of hip fracture and lower bone mineral density in older adults with subclinical hyperthyroidism (SHyper) and subclinical hypothyroidism (SHypo).[br][bold]Methods:[/bold] Incident hip fracture was analyzed in US community-dwelling men (n=2,171) and women (n=2,765) aged [ge]65 y enrolled in the Cardiovascular Health Study. Cox proportional hazard models were used in sex-stratified analyses to examine the relationship between baseline thyroid status and incident hip fracture. All models were adjusted for age, race, weight, physical activity, alcohol use, smoking, oral steroid use, and estrogen use (in women). Initial models excluded participants taking thyroid medications, and a second set included all participants with adjustment for thyroid medication use. Cross-sectional analyses of hip bone mineral density were performed 598 men and 719 women who had DXA scans. All analyses were repeated in a subset of participants who had a second set of thyroid measurements to define persistent, transient, or progressive SHypo.[br][bold]Results:[/bold] There were 564 cases of incident hip fracture (469 euthyroid, 84 SHypo, 11 SHyper) over a median follow-up of 12 years. We found no association of thyroid status with incident hip fracture in men or women. Although the point estimates suggested higher fracture risk in men with SHyper than in euthyroid men (HR 1.78, 0.56-5.66), this was based on few events, was further attenuated in models including thyroid hormone users (HR 1.18, 0.77-1.80), and was not found in women (HR 0.91, 0.69-1.20). Baseline SHypo was not associated with higher fracture risk in either men (HR 1.27, 0.82-1.95) or women (HR 1.11, 0.55-2.25), nor was persistent SHypo in men (HR 1.09, 0.57-2.10) or women (HR 0.79, 0.52-1.21). There was no difference in total hip or femoral neck bone mineral density in men and women, comparing SHypo and SHyper to euthyroid using a single timepoint or in models of persistence, and when analyzing TSH as a continuous variable.[br][bold]Conclusions: [/bold]We found no difference in the incidence of hip fracture or lower bone density in men and women with baseline or persistent SHypo. There may be a higher incidence of hip fracture in men with SHyper, but power was limited to assess this risk. We find no evidence for a clinically significant impact of subclinical thyroid dysfunction on hip fracture in this population.[br][br]Sources of Research Support: R01 AG032317 to ARC.[br][br]Nothing to Disclose: MCG, AMA, JSL, JAR, ARC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1231 191 1436 SUN-409 PO39-02 Sunday 1233 2012


1230 ENDO12L_SUN-410 POSTER SESSION: Thyrotoxicosis, TSH Administration [amp] Thyroid Cell Biology (1:30 PM-3:30 PM) Bedside Ultrasound Use in Graves Disease Yannis S Guerra, Juanito U Uy, Aarti Manchanda, Leon Fogelfeld John H Stroger Jr, Hospital of Cook County, Chicago, IL; Rush University Medical Center, Chicago, IL Introduction[br]Radioactive iodine therapy(RAI) dosing in Graves[apos]s disease(GD) requires an estimate of the thyroid volume usually done by clinical exam or thyroid ultrasound(US). We measured the correlation between these techniques and also evaluated post RAI volumetric changes using bedside US.[br]Methods[br]Subjects treated with methimazole until a week prior to RAI therapy had thyroid physical exam volume estimates by clinical endocrinologists (PEVOL). The RAI dose was calculated using PEVOL. This was compared with a bedside thyroid gland US volume (USVOL) generated by the US calculator performed by 2 researchers (blinded to each other results) on the day of RAI. The average value from both measurements was used for all calculations. Repeat USVOL was performed on patient follow up by the same researchers. The pre and post RAI volumes (PEVOL and USVOL) were correlated to the pre and post RAI hormone tests.[br]Results[br]20 GD subjects (14 females; Age = 44[plusmn]14.7 years) underwent thyroid volume and hormone measurement. 18 had post RAI USVOL and 19 had FT4 levels on follow up. Time of follow up USVOL after RAI was 161[plusmn]14 days.[br]The correlation coefficient of the USVOL between the two researchers per diameter measures went from 0.76 to 0.94. Pre-RAI Mean PEVOL was higher than USVOL (47.9[plusmn]24 gr vs. 31.8[plusmn]15gr) but correlated with each other (r=0.6, p=0.006). The USVOL at follow- up was 6.7[plusmn] 4.6. The decrease in volume was -6.2[plusmn]6 ml/month and it correlated (r=0.5, p=0.05) with RAI dose expressed as mCi/gram USVOL. The follow-up USVOL was lower in responder to therapy at that time point compared with non-responders (4.7 vs. 10.7 gr ,p=0.004).[br]Mean RAI dose given was 19.7[plusmn]6 mCi equivalent to 0.48 mCi/gr of thyroid tissue by PEVOL and 0.79 mCi/gr by USVOL.[br]Pre RAI volume correlated with the RAI dose (r= 0.6 for PEVOL and r=0.68 for USVOL, p[lt]0.01 both) but only the ratio of RAI dose per USVOL correlated with both post RAI volume (r=-0.51, p=0.03) and TSH level (r=0.56, p=0.01). [br]Post RAI volume was strongly correlated with the percent change in the FT4 level (r=0.74, p=0.001) but not with the absolute change in FT4 levels (r=0.365, p=0.15).[br]Conclusion[br]Among GD patients, physical exam correlates with US measurements but overestimates the volume by 30%. Use of US thyroid volume assessment at baseline for dose estimates and subsequently for follow-up to gauge response to therapy may be a useful tool. Prospective larger studies are needed to establish its value as predictor of response.[br][br]Nothing to Disclose: YSG, JUU, AM, LF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2162 191 1437 SUN-410 PO39-02 Sunday 1234 2012


1231 ENDO12L_SUN-411 POSTER SESSION: Thyrotoxicosis, TSH Administration [amp] Thyroid Cell Biology (1:30 PM-3:30 PM) Atenolol Is Better Than Propranolol for Control of Thyrotoxicosis Manifestations Saad Sakkal Metabolic Care Center, Aleppo, Syrian Arab Republic Objective: Atenolol is a beta blocker with less cerebral effects than propranolol. However, propranolol has been the commonly used beta blocker in controlling thyrotoxicosis manifestations for many years.We prospectively compared their effects on clinical manifestations.[br]Methods: We report the results of the first 100 patients who were assigned alternatively to either drugs.Effects include Pulse, Bp, weight, thyroid tests and subjective feeling palpitation, tired, hot, headache, sleep.[br]Results: There was no significant difference in thyroid tests (T4:2.1VS1.8), sleep or weight. However patients reported with Atenolol less feeling of palpitation, fatigue, hot or headache/cerebral symptoms. Pulse and Bp control was better with Atenolol.[br]Average means differences were: Bp 8/7(126/76 with Propranolol (Pr), 118/69 with Atenolol (At). Pulse: 22(94 with PR, 72 with At). Feeling palpitation: %30(%45 with Pr, %15 with At). Fatigue: 2/5(4/5 with Pr, 2/5 with At). Feeling hot 2/5(3/5 with Pr, 1/5 with At). Cerebral Symptoms: %32(%35 with Pr, %12 with At). And finally expense:$12 or %20($60 with Pr, 48 with At). All differences were significant at 0.001 P value or more.[br]Conclusion: Atenolol is much better choice in controlling thyrotoxicosis manifestations than propranolol. It has better control of Pulse, Bp, palpitation, fatigue, warm feeling and cerebral symptoms probably because of its longer half-life and less central cerebral penetration.[br][br]Nothing to Disclose: SS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 152 191 1438 SUN-411 PO39-02 Sunday 1235 2012


1232 ENDO12L_SUN-412 POSTER SESSION: Thyrotoxicosis, TSH Administration [amp] Thyroid Cell Biology (1:30 PM-3:30 PM) The Effect of Thyroid Scintigraphy on Thyroid Function Test among the Patients with Subclinical Hyperthyroidism Paul Joon Kim, Narmada Movva, Negar Niknam, Negin Niknam, David Michael Reich, Issac Sachmechi Queens Hospital Center/Mt Sinai School of Medicine, Jamaica, NY Background/Aim: Thyroid Scintigraphy (TS) is often ordered for the evaluation of subclinical hyperthyroidism (SH). The purpose of this study is to investigate the possibility of TS normalizing thyroid stimulating hormone level (TSH) in significant number of patients with SH.[br]Method: Our study subjects were screened out of a total of 520 patients who had TS in the past 5 years at our institution. A total of 175 patients with SH were identified. From these patients, 105 patients (male: 19, female: 86) met our inclusion and exclusion criteria which might have affected TSH during the data collection period and were chosen as the study group. The patients[apos] TSH levels prior and post TS up to one year were collected and analyzed for any significant changes.[br]Result: 31 (29.5%) patients[apos] TSH levels normalized after TS (response group), 23 (21.9%) patients[apos] TSH levels showed variable responses with fluctuating TSH levels between normal limits and SH range (variable group), and 51 (48.6%) patients[apos] TSH levels remained within SH range (non-responsive group). The responsive group[apos]s mean TSH changed from 0.3 to 0.903 (mIu/ml) with p[lt]0.01 post TS. The variable group[apos]s mean TSH changed from 0.26 to 0.371 (mIu/ml) with p[lt]0.01 post TS. The non-responsive group[apos]s mean TSH remained unchanged from 0.198 to 0.19 (mIu/ml) post TS with a non-significant (NS) p value.[br]Discussion: This study demonstrates that about 30% of patients with SH normalized their TSH after having TS. A possible explanation for the different response between groups could be age and TSH level right before TS (pre TS). The responsive group[apos]s mean age (51.5 years) was younger than the variable group[apos]s mean age (62.9 years) with p[lt]0.01 and younger than the non-responsive group[apos]s mean age (56 years) with NS p value. The non-responsive group[apos]s TSH pre TS (0.2 mIu/ml) was lower than the variable group[apos]s TSH pre TS (0.263 mIu/ml) with a p[lt]0.05 and lower than the responsive group[apos]s TSH pre TS (0.257 mIu/ml) with a NS p value.[br]Conclusion: Based on our research analysis, TS can normalize TSH levels in significant number of patients with SH. We suggest to consider to follow up TSH levels at least a year post TS if a patient with SH is younger than 51 year old and has pre TS TSH level higher 0.25 (mIu/ml) to avoid iodine 131 ablation therapy.[br][br]Nothing to Disclose: PJK, NM, NN, NN, DMR, IS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2348 191 1439 SUN-412 PO39-02 Sunday 1236 2012


1250 ENDO12L_SUN-430 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Reversible Dilated Cardiomyopathy Associated with Graves Disease Mansi Mehta, Kenneth H Hupart NuHealth, Nassau University Medical Center, East Meadow, NY; St John[apos]s University, Queens, NY; Albert Einstein College of Medicine, Bronx, NY A 55-year-old African American female was admitted to the hospital with worsening shortness of breath and bilateral lower extremity edema. Echocardiography showed a left ventricular ejection fraction of 25% with global hypokinesis and dilatation of all four cardiac chambers. Work-up, including cardiac catheterization, did not reveal any coronary disease. Thyroid testing revealed a suppressed serum TSH with elevation of T3 (279 ng/dL) and Free T4 (2.16 ng/dL) along with positive thyroid peroxidase, thyroglobulin, and thyroid stimulating antibodies. She denied any personal or family history of thyroid and cardiac disease. The patient was diagnosed with new onset dilated cardiomyopathy and systolic heart failure likely secondary to autoimmune thyroid disease. A 24-hour radio-iodine uptake of 89% confirmed the presence of Graves[apos] disease. She was treated for heart failure and was started on methimazole and beta-blockers for her thyrotoxicosis. On Methimazole treatment, the patient[apos]s T3 and Free T4 reached the mid-normal range and TSH normalized. A repeat echocardiogram performed six months after presentation showed complete resolution of heart failure with a normal ejection fraction and no wall motion abnormalities.[br]Dilated cardiomyopathy has been described in association with thyrotoxicosis, Graves[apos] disease, toxic multinodular/uni-nodular goiter and hypothyroidism. Reversible thyrotoxic cardiomyopathy is a rare event that has mostly been associated with thyrotoxicosis caused by Graves[apos] disease. This has lead to speculation that the dilated cardiomyopathy seen in patients with Graves[apos] disease may be due to underlying autoimmunity rather than thyrotoxicosis per se. This has been fueled by the observation of cardiomyopathy preceding the diagnosis of thyrotoxicosis, reports of lymphocytic myocarditis in some patients, and the reported identification of TSH receptor mRNA in human myocardial tissue.[br]The clinical course of Graves[apos] orbitopathy has been observed to worsen with radio-iodine therapy which can induce auto-antibody production. Thyroidectomy, which can promote a fall in TSH receptor auto-antibody titers, may improve orbital disease. The prospect that the cardiomyopathy in patients with Graves[apos] disease shares a common pathogenesis with orbitopathy suggests that thyroidectomy should be the preferred therapeutic approach in these patients.[br][br]Nothing to Disclose: MM, KHH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1820 192 1457 SUN-430 PO56-02 Sunday 1254 2012


1251 ENDO12L_SUN-431 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Treatment of Severe Graves Ophthalmopathy with a PPAR-[gamma] Antagonist and Cylooxygenase-2 Inhibitor Janete Moura, Lidia Yuri Mimura, Walter Bloise S[atilde]o Paulo University-School of Medicine, S[atilde]o Paulo, Brazil [bold]Introduction[/bold] - We have demonstrated that the expression of PPAR-[gamma] is increased in the retro-ocular adipose tissue in active phase of Graves[apos] ophthalmopathy [bold](1)[/bold]. According to the new trends of target therapy, our previous study showed that PPAR-[gamma] antagonist and cyclooxygenase-2 inhibitor (sodium diclofenac) was effective to treat patients with mild to moderate model of this ocular disease [bold](2)[/bold]. The use of this drug to treat patients with sight threatening was doubtful.[br][bold]Background[/bold] - The aim of this case report is to show the benefits of sodium diclofenac in a patient with severe Graves[apos] ophthalmopathy.[br][bold]Clinical case[/bold] - A 44-year-old woman was admitted in the Thyroid Unit complaining of ocular pain and decrease of vision in the last 2 months. One year before she noticed protrusion and pain in her eyes together with symptoms of hyperthyroidism. Due to the sub acute onset of vision involvement we decided to try our new therapy. The hyperthyroidism was clinical and laboratory fairly controlled with 10 mg daily of methimazole.[br]The congestive eyes were classified by the clinical activity scores (CAS) as 7 and the proptosis measured by Hertel exopthalmometer were 20 mm and 22 mm of the right and left eye respectively. Visual acuity evaluated by Snellen chart was 0.9 and 0.7 of right and left eye respectively. The left eye showed narrowing of the visual field. Sodium diclofenac was administered by intramuscular injection of 75 mg every 12 hours in the first week followed by 50 mg orally every 12 hours for 12 months.[br][bold]Results[/bold] - After one week of treatment the visual acuity by Snellen chart was 1.0 in both eyes and the left visual field was normal 1 month after checking. No adverse effects were found and the blood and renal tests remained normal checked every 2 months. After the drug withdrawal there was no relapse of the ophthalmopathy and 6 months after she presented ocular signs classified as CAS 3. The hyperthyroidism persisted requiring methimazole treatment.[br][bold]Conclusions[/bold] - Sodium diclofenac which is a target therapy may be a good, safe and less expensive option to treat selected patients with severe Graves[apos] ophthalmopathy.[br][br]1)L[iacute]dia Y. Mimura, Sandra M.F. Villares, M[aacute]rio L.R. Monteiro, Isabel C. Guazzelli, and Walter Bloise. Peroxisome Proliferator-Activated Receptor-[gamma] Gene Expression in Orbital Adipose/Connective Tissues is Increased During the Active Stage of Graves[apos] Ophthalmopathy Thyroid. September 2003, 13(9): 845-850. 2) Bloise W, Mimura LY, Moura J, Nicolau W. Treatment of mild to moderate Graves[acute] ophthalmopathy with sodium diclofenac: a pilot study. Brazilian Archives Endocrinol Metabol, 2011;55(9): 692-695.[br][br]Nothing to Disclose: JM, LYM, WB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 255 192 1458 SUN-431 PO56-02 Sunday 1255 2012


1252 ENDO12L_SUN-432 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Development of Polyarthritis during Methimazole Therapy for Graves Disease Ben O[apos]Donnell, Hilary Whitlatch Brown University Rhode Island Hospital, Providence, RI [bold]Introduction[/bold]: Antithyroid drugs (ATDs) are known to cause a variety of minor and major adverse reactions, ranging from rash to hepatic failure. While arthralgias can occur in up to 5% of patients on ATDs true arthritis is relatively rare, estimated to occur in 1-2% of patients. This case summarizes the evaluation and course of a patient admitted with the antithyroid arthritis syndrome.[br][bold]Clinical Case[/bold]: A 51 year-old male with Graves[apos] disease presented with 4 days of progressive joint pain, inability to walk, and fever. Six weeks prior, he was started on Methimazole 3 times per day, which was decreased to twice daily dosing when the symptoms first began. Joint pain began in his right third digit; then progressed to involve his bilateral shoulders, left foot, and knee. In the ER, he was noted to have a moderate effusion of the left knee. Arthrocentesis yielded cloudy yellow fluid with 49,500 nucleated cells, 93% neutrophil predominance, and negative gram stain. Serum WBC was normal at 6.5x10[sup]9[/sup]/L. The patient had a mildly reactive ANA at 1:80 (ref [lt]1:40), with a nucleolar pattern, and negative anti-Histone antibody (ref [lt]1). ANCA was negative (ref [lt]1:20). Thyroid studies included: TSH [lt]0.03 mIU/mL (0.35-5.5), free T4: 2.19 ng/dL (0.8-1.8), and total T3: 248 ng/dL (80-180).[br]At admission, Methimazole was stopped; he was placed on Propranolol which was titrated to control adrenergic symptoms. Once the gram stain from the arthrocentesis was negative, he was started on glucocorticoids. He remained afebrile throughout his hospital stay, and was able to ambulate without pain within 24 hours of starting Prednisone. Upon discharge he completed the course of Prednisone and had definitive treatment of Graves[apos] disease with radioactive iodine.[br][bold]Conclusion[/bold]: The antithyroid arthritis syndrome consists of arthritic symptoms involving one or more small, medium, or large joints that can develop within 8 weeks of starting ATD. In the event that arthritis develops, the medication should be stopped, and the alternative thionamide should be avoided due to high cross-reactivity. Some patients, depending on severity, can be managed with NSAIDs alone, while others may require glucocorticoids. Symptoms can last 3-4 weeks even after stopping the antithyroid medication, and definitive treatment of hyperthyroidism should be sought either with radioactive iodine or surgery.[br][br]Nothing to Disclose: BO, HW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2066 192 1459 SUN-432 PO56-02 Sunday 1256 2012


1253 ENDO12L_SUN-433 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Thyrotoxic Hypokalemic Periodic Paralysis, Case Presentation and Review Muhammad Houri Sparks Health, Fort Smith, AR Case: 23-year-old, Asian male was visiting his parents when he started complaining of weakness in all extremities, this progressed over 24 hours. When he started having shortness of breath he was brought to ER by EMS. On presentation he had SVT followed by V tach.Exam revealed an anxious looking diaphoretic male who was unable to move his extremities. He has a diffuse goiter, but no exophthalmos or dysmorphic features.[br]Tachyrhythmia was treated with Cardizem, cardioversion then he was amiodarone drip for less than60 minutes. Potassium was 1.1mg/dl, Mg 1.3, Both were replaced. He was admitted to ICU and started on Inderal. Review of system was significant for 3 episodes of weakness with spontaneous resolution, 30 lbs weight loss and heat intolerance.[br]Thyrotoxic hypokalemic periodic paralysis(PP)was suspected.[br]TSH 0.01, FT4 =4.15 (0.9-1.8 ng/dL) and FT3 =11.5 (2.3-4.1 ng/mL).Thyroid uptake done using 75 microcu of I-123.4 and 24 hour uptake were 27% and 17.4% respectively[br]After completing the four hour uptake, Tapazole 20 mg 3 times daily was started. Uptake at 4 hours was elevated consistent with Graves[apos] disease. The 24-hour uptake was lower likely because he did receive amiodarone drip for about 30-60 minutes, and Tapazole starting AFTER the 4 hour uptake was completed. [br]TSI was 210 (nl [lt] 140) TPO Ab 364 iu/mL, (nl [lt] 35)[br]He had 3 episodes of painless weakness with spontaneous resolution in the last year. The episodes usually happen when he visits his parents (where he eats Asian food rich in carbohydrate, mainly rice and noodles).[br]No known FHx of periodic paralysis or hyperthyroidism.[br]His paralysis resolved few hours after admission.Potassium was normalized less than 24 hours.Within 48 hours he was back in normal sinus rhythm and was able to ambulate without difficulty[br]Discussion:Periodic Paralysis is a rare disease manifests as episodes of painless muscle weakness., usually precipitated by exercise, fasting, or high-carbohydrate meals. PP is classified as hypokalemic and hyperkalemic depending on potassium level which trigger the episodes. Most cases of PP are inherited with an autosomal dominant pattern. However, acquired cases of hypokalemic PP have been described in association with hyperthyroidism. The incidence of Thyrotoxic PP is significantly higher in males of Asian descent.[br]Replacing potassium causes rapid resolution of weakness. And treating the underlying hyperthyroidism usually prevents further attacks.[br][br]Nothing to Disclose: MH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2372 192 1460 SUN-433 PO56-02 Sunday 1257 2012


1254 ENDO12L_SUN-434 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Preoperative Plasmapheresis Treatment for Resistant Thyrotoxicosis Faysal Albaalbaki, Khalid Saleem, Omar Saleem, Valentin Zaharia, Mohamad H Horani Banner Baywood Hospital, Mesa, AZ; Chandler Hospital, Chandler, AZ; AKDHC, Phoenix, AZ; Alsham Endocrinology, Chandler, AZ A 78 year-old female with a history of osteoporosis presented with right hip fracture status post fall.[br]She has a history of frequent palpitations and weight loss. On admission, a CT head and neck showed large heterogeneous thyroid with multiple nodules. She was diagnosed with paroxysmal atrial fibrillation and pulmonary emboli and admitted to a monitor floor. Her hip surgery was held.[br]Thyroid function test revealed severe thyroid toxicosis with TSH less than 0.01. Free T4 levels were greater than 8 (normal levels.9 [ndash] 1.7 pg/mL). Free T3 levels were 19.8 (2.0 [ndash] 4.8 pg/mL). 25 Hydroxy vitamin D level was 28.2 ng/ml[br]She was diagnosed with graves[apos]s disease and started on methimazole, as well as propranolol to control her tachycardia. Her thyroid function was monitored throughout her initial hospital stay, and no response to medical treatment was achieved. Her laboratory testing showed persistent elevation in free T4 and free FT3, despite adding steroids to her regimen. Plasmapheresis was suggested preoperatively. Treatment with plasmapheresis induced more than 50 % decrease in Free T4 (from 6.2 pg/ml to 3.0 pg/ml) free T3 also normalized (from 9.3 pg/ml day of plasma phresis to 3.2 pg/ml after 24 of plasma phresis). we obsreved decline in 25 vitamin D value more than 30 % one day after plasma phresis. (from 28.2 ng/ml to 18.3 in 48 hour).[br]Patient underwent right hip surgery with no complications and in 4 days later she had total thyroidectomy. The thyroid was removed because the patient was very resistant to tabazole, which was administered on the maximum dosage for 10 days. Radioactive iodine ablation was not viable option due to urgency of hip surgery and risk of thyroid storm. We observe similar decline in vitamin D levels in 12 other pateints recieving plasma phresis for other etiologies. we don,t know if plasmaphresis affected lab assay or unknown binding protein.[br]This case illustrates the effectiveness and benefit of preoperative plasmapheresis treatment for severe and resistant thyrotoxicosis. Recognition of this treatment option is very important in the hospital setting by decreasing hospital stay awaiting euthyroidism state not achieved by traditional medical treatments. Preoperative treatment is vital to avoid possibility of inducing a thyroid storm during surgery due to uncontrolled thyrotoxicosis. the decline in vitamin D level with plasma phresis deserves further investigation.[br][br]Nothing to Disclose: FA, KS, OS, VZ, MHH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 28 192 1461 SUN-434 PO56-02 Sunday 1258 2012


1255 ENDO12L_SUN-435 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Atypical TSHoma Presenting with Prolonged Non-Progressive Central Hyperthyroidism, with Delayed Appearance on MRI Pituitary Imaging Yin Chian Kon Tan Tock Seng Hospital, Singapore, Singapore [bold]Introduction[/bold]:TSHomas typically present with central hyperthyroidism due to a TSH-secreting pituitary macroadenoma. A female with prolonged central hyperthyroidism, non-progressive over 5 years, not due to thyroid hormone resistance, with equivocal presence of pituitary microadenoma, is presented.[bold]Case Report[/bold]:34-year-old female was referred in May 2006 for hand tremors associated with elevated fT4 2.5 ng/dl (1.0-1.8) and non-suppressed TSH 2.62 mIU/L (0.27-4.2). She denied any change in weight or appetite nor heat intolerance. She had occasional palpitations for a few years. Menses slight change from 4-5/30 to 38-39 day cycle. No visual disturbance. She had Ht 163.6 cm, Wt 52.5 kg, BMI 19.7 kg/m2, BP 114/59 mmHg, HR 120/min regular. Mild tremors, no goiter nor extrathyroidal features of Graves[apos] disease. Repeat TFT showed fT4 26 pM (8-21) TSH 2.01 mIU/L (0.34-5.6) (Beckman-Coulter Dxl), fT4 27 pM (10-23), TSH 2.14 mIU/L (0.45-4.5) (Siemens Centaur). Anti-TPO, anti-thyroglobulin and TRAb antibodies were negative. IV TRH (200 ucg) stimulation revealed raised basal fT4 28 pM, fT3 9.8 pM (4.3-8.3), SHBG 166 nM (20-122) and non-stimulated TSH: 0 min 1.62; 15 min 3.29; 30 min 2.98; 60 min 2.34 mIU/L. However, dynamic MRI pituitary scan was normal, with no ectopic focus seen. Werner[apos]s protocol (TRH stimulation-T3 suppression over 9 days) in July 2010 excluded the presence of thyroid hormone resistance, and was compatible with autonomous central hyperthyroidism. Repeat dynamic pituitary MRI on Sep 2010 and Oct 2011 revealed a stable, small, 4 mm x 3 mm x 4 mm area of differential enhancement in the right anterior pituitary lobe. The patient refused TSS nor octreotide treatment, and remained clinically well on low-dose beta-blockade. Her serial TFTs from May 2006 to Oct 2011 (range): fT4 22-30 pM, fT3 6.4-9.8 pM, TSH 1.62-3.29 mIU/L, SHBG 135-166 nM[bold]Discussion[/bold]:TSHomas are increasingly being recognized at the stage of. microadenomas, due to use of ultrasensitive TSH assays and increasing awareness (1). Rougemont et al have reported a male patient with long-standing central thyrotoxic-induced severe osteoporosis where the TSHoma was not evident on MRI until 5 years after presentation (2). This case illustrates that TSHoma may not initially be radiologically evident despite causing prolonged mild central hyperthyroidism.[br][bold]Conclusion[/bold]:TSHomas may present atypically with prolonged mild central hyperthyroidism and initially negative MRI imaging.[br][br](1) Socin HV et al., European Journal of Endocrinology 2003;148:433. (2) Rougemont A et al. J Bone Miner Metab 2009;27:513.[br][br]Nothing to Disclose: YCK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1191 192 1462 SUN-435 PO56-02 Sunday 1259 2012


1256 ENDO12L_SUN-436 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Case Report: Delayed Presentation of Thyroid Storm after Radioiodine Therapy Yin Htwe Oo, Nathaniel Winer, MaryAnn Banerji State University of New York (SUNY) Downstate Medical Center, Brooklyn, NY Introduction[br]Radioiodine (RAI) therapy is widely used for treatment of Graves[apos] disease (GD). A RAI dose of 20 m Ci is reported to result in hypothyroidism in 75 to 90% of cases while others note that hyperthyroidism can be treated with 8 mCi of RAI. However, up to 28% remain hyperthyroid and 44% become hypothyroid. Delayed exacerbation of GD by RAI therapy is relatively rare (0.4%). Its mechanism may involve autoimmunity. Here, we report a case of GD presenting with thyroid storm five weeks after receiving RAI.[br]Case report[br]A 53 year-old man with history of hypertension was diagnosed with GD after he presented with extreme weakness and weight loss. Thyroid function tests (TFTs) showed TSH of 0.008 (0.35-5.5mIU/L), free T4 (FT4) of 3.19 (0.89-1.76ng/dl) and negative thyroid peroxidase antibody (TPO). Thyroid scan showed a homogenous gland with a 64.5% uptake at 24 hours. He received a standard dose of 8mCi of RAI-131 seventeen days after the scan with no anti-thyroid drug treatment prior to RAI therapy. Five weeks later, he was hospitalized with tiredness, palpitations and fever without neck pain. Burch and Wartofsky score was 45; highly suggestive of thyroid storm. TFTs revealed TSH of [lt]0.004, FT4 of 9.35, and total T3 of 773(60-181ng/dl).TPO was positive (2,787 (Normal[lt]=60)). He improved with propylthiouracil, potassium iodide, steroid and propranolol treatment.[br]Discussion[br]The development of thyrotoxicosis after RAI therapy is believed to be mediated by two different mechanisms: a transient increased release of thyroid hormones due to radiation thyroiditis and the rare development of GD due to formation of antibodies to thyroid-associated antigens released from damaged follicular cells. Immunologically-mediated exacerbation of hyperthyroidism differs from radiation-induced thyroiditis by late onset of symptoms, elevated thyroid antibodies and increased RAI uptake. Immune-mediated paradoxical exacerbation of GD with a low RAI dose (e.g.,8 mCi) has been reported.[br]Conclusion[br]The exacerbation of thyrotoxicosis after RAI therapy should be anticipated in GD especially in patients with high RAI uptake. The data suggest that GD patients with high radioiodine uptake may benefit by establishing euthyroidism prior to RAI therapy, and giving a sufficiently large dose of RAI. Close monitoring with TFTs, antibodies titers and, if necessary, restarting antithyroid drugs 7 days after RAI therapy may minimize the risk of developing paradoxical exacerbation of GD.[br][br]Nothing to Disclose: YHO, NW, MB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1092 192 1463 SUN-436 PO56-02 Sunday 1260 2012


1257 ENDO12L_SUN-437 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Psychosis in Hyperthyroidism Paulette David Nacpil, Cherrie Mae Sison University of the Philippines-Philippine General Hospital, Manila, Philippines Background: The most common psychiatric syndromes in thyrotoxicosis are anxiety and mood disturbances. Psychosis, however, is a rare complication. Here is a report of such a case.[br]Case report: A twenty-six year old female presented with a thirteen-year history of palpitations, easy fatigability and anterior neck mass. Patient denies any comorbid conditions. No family history of psychiatric illness. On March 2011, she was admitted due to logorrhea, auditory hallucinations, delusion of persecution and frequent attention calling from family members, and suicidal ideations. Clinically she was thyrotoxic with a diffuse goiter. FT4 39.6 pmol/L(11-24) and TSH [lt]0.005 [micro]IU/L (0.3-2.8). There was no evidence of infection. Diagnosis of thyrotoxic psychosis was made, and was started on Propylthiouracil 150 mg q8 and Propranolol 40mg BID.[br]After her marked improvement, she was discharged on Propylthiouracil, Propranolol, Olanzapine and Clonazepam. All her psychiatric symptoms gradually resolved and weaned off from Olanzapine and Clonazepam. During her regular follow up, there was note of occasional episodes of agitation. Six months later, after euthyroidism was achieved, patient underwent radioactive ablation, with complete resolution of her psychiatric symptoms.[br]Conclusion: Thyrotoxicosis may be a precipitant of acute psychosis. This can be promptly controlled with the use of anti-thyroid drugs. Concomitant psychotropic drugs maybe indicated if the symptoms are severe. In cases of treatment failure, a more radical approach such as radioactive iodine ablation and thyroidectomy maybe considered.[br][br]Nothing to Disclose: PDN, CMS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2369 192 1464 SUN-437 PO56-02 Sunday 1261 2012


1258 ENDO12L_SUN-438 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) A Case of Relapse of Graves Disease Presenting as Free T3 Thyrotoxicosis in the Postpartum Period Madiha Khan, Sara Ahmadi, Randall Urban The University of Texas Medical Branch at Galveston, Galveston, TX [bold]Introduction:[/bold][br]Relapse of Graves[apos] disease associated with an elevated free T4 occurs commonly in the postpartum period. However, postpartum free T3 thyrotoxicosis, which is described in the setting of normal serum total and free T4 level in patients with Graves[apos] disease is not common. The prevalence of free T3 thyrotoxicosis as a result of relapse of Graves[apos] disease in the postpartum period is unknown.[br][bold]Clinical case:[/bold][br]A healthy 29-year-old female with a past medical history of Graves[apos] disease in remission presented to our clinic for regular follow-up 4 months postpartum after her second pregnancy. She complained of symptoms of anxiety, insomnia, palpitations, heat intolerance and excessive sweating. Physical exam revealed an anxious and hyperactive female. She was tachycardiac, hypertensive and had ocular features of Graves[apos] with hyperreflexia, warm and moist hands with tremors. Laboratory work-up revealed undetectable TSH, normal free T4 of 1.97 ng/dL (normal: 0.78-2.20) and an elevated free T3 of 8.3 pg/mL (normal: 2.4-4.2). The patient refused to take PTU and was started on Propranolol with the diagnosis of postpartum relapse of Graves[apos] disease leading to free T3 thyrotoxicosis.[br]The patient was diagnosed with Graves[apos] disease in 2006 and was treated with PTU for almost one year, which was stopped before her first pregnancy once she became euthyroid. Thyroid function tests remained stable during pregnancy despite not being on any treatment. Eight months postpartum, she developed a relapse of Graves[apos] disease associated with exopthalamos. The patient was restarted on PTU and five months later, PTU was switched to Methimazole to improve patient compliance. Methimazole was stopped 10 months later once the patient became euthyroid. She had her second pregnancy after being in remission for 7 months. The patient also remained euthyroid and did not require treatment during the second pregnancy. She became hyperthyroid again four months postpartum when she was seen in our clinic.[br][bold]Conclusion:[/bold][br]In reviewing the literature, cases of free T3 thyrotoxicosis are rare. This case demonstrates that we should consider checking free T3 in Graves[apos] disease patients with normal free T4 if they present with signs and symptoms of hyperthyroidism.[br][br]Nothing to Disclose: MK, SA, RU 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 706 192 1465 SUN-438 PO56-02 Sunday 1262 2012


1259 ENDO12L_SUN-439 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Thyroid Storm Presenting as Severe Cholestasis Subsequently Leading to Multiorgan Failure Ivania Maritza Rizo, Crystal Bowden University of Alabama at Birmingham, Birmingham, AL [bold]Introduction [/bold][br]Thyroid storm is rare, life-threatening condition, usually occurring in patients with preexisting thyrotoxicosis. We discuss a patient with Graves[apos] disease who presented with severe cholestasis and subsequent multiorgan failure treated with emergent thyroidectomy.[br][bold]Clinical Case[/bold][br]A 32 year old woman presented to a community hospital with jaundice, abdominal pain, vomiting, and diarrhea. She carried a diagnosis of Graves[apos] disease and atrial fibrillation, but was not receiving treatment for these conditions. Admission laboratory data was significant for cholestasis. Work up excluded viral hepatitis, autoimmune hepatitis, Wilson[apos]s, negative toxicology and per imaging no signs of biliary obstruction. Thyroid studies showed thyroid stimulating hormone 0.006 mU/L (n 0.45 -4.5 mU/L), serum total T4 24.2 mcg/dL (n 4.5- 12mcg/dL) and T3 [gt] 651 ng/dL (n 83-200ng/dL). Thyroid ultrasound and uptake were consistent with Graves[apos] disease. Liver biopsy demonstrated bile stasis with lobular inflammation and spotty hepatocellular necrosis. Her clinical status deteriorated and required transfer to our center for liver transplant evaluation.[br]Upon transfer, the patient was agitated and exam revealed exophthalmos, jaundice, enlarged nontender thyroid without bruit, tachycardic irregularly irregular heart rhythm, and resting tremor. Her laboratory studies were significant for a total bilirubin of 32 mg/dL (n 0.4-1.4mg/dL) and mild transaminitis. Cardiac functional assessment was essentially normal. She developed worsening cholestasis, encephalopathy, hypoglycemia, and acute kidney injury requiring hemodialysis. On day 10 following transfer, the patient was begun on methimazole 20mg daily. Her transaminases immediately trended upward with evidence of worsening synthetic liver function. Methimazole was discontinued, and the patient was taken for total thyroidectomy. Following surgery, hepatic and renal function slowly improved and she made a full recovery. Thyroid pathology was consistent with Graves[apos] disease.[br][bold]Conclusion[/bold][br]Although hyperthyroidism is associated with liver function abnormalities, severe cholestasis is a rare presenting feature of thyroid storm. Multiple cases have reported resolution of cholestasis after initiation of thionamides in the absence of another liver disease; however our patient did not tolerate methimazole. This case empthasizes that if thionamides are ineffective, consideration should be given to total thyroidectomy.[br][br]Nothing to Disclose: IMR, CB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 811 192 1466 SUN-439 PO56-02 Sunday 1263 2012


1260 ENDO12L_SUN-440 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) New Onset Graves Disease after Radioactive Iodine Treatment of Toxic Multinodular Goiter Spyridoula Maraka, Danae Delivanis, Pooja Luthra University of Connecticut Health Center, Farmington, CT [bold]Background:[/bold] Development of Graves[apos] disease in patients with toxic multinodular goiter after radioiodine treatment (RIT) has been described at a rate of 1% to 5%.[br][bold]Clinical Case:[/bold] A 39-year woman had a 7-year history of toxic multinodular goiter with subclinical hyperthyroidism. Initial tests revealed: suppressed TSH (0.01 mIU/l, n=0.4-4.5 mIU/l) with FT4 of 1.2 ng/dl (n=0.8-1.8 ng/dl) and FT3 of 3.8 pg/ml (n=2.3-4.2 pg/ml). TSI was 105% (n[lt]125%) and anti-TPO antibodies level was 1.9 IU/ml (n=0.0-2.0 IU/ml). A thyroid ultrasound revealed multiple subcentimetric nodules bilaterally and normal echogenicity. A thyroid radioiodine uptake was normal at 20% at 24 hours (n=10-30%). Thyroid was mildly enlarged, more at the right lobe, with mildly non-uniform tracer uptake. The patient was subsequently treated with 12.5 mCi of I-131 without any side effect. Three months later she experienced palpitations, heat intolerance, and an unintentional 8-pound weight loss. Repeat tests showed TSH less than 0.01 mIU/l with FT4 of 5.9 ng/dl and total T3 more than 800 ng/dl (n=76-181 ng/dl). TSI was 150%. The patient was diagnosed with Graves[apos] disease and was started on methimazole.[br]It has been suggested that the development of post radioactive iodine immunogenic hyperthyroidism may be caused by exacerbation of preexisting Graves[apos] disease, that was not diagnosed due to previous insensitive TSH-receptor antibodies (TRAbs) assays. Massive destruction of thyroid follicular cells is observed following RIT and may expose epitopes to the immune system that are processed by antigen presenting cells thus inducing autoimmunity. Reported risk factors for the development of postradioiodine immunogenic hyperthyroidism include: TRAbs level in the upper range of normal, elevated anti-TPO antibodies, a diffuse hypoechoic pattern, and a diffuse scintigraphic uptake pattern. Interestingly, in our patient no signs of autoimmunity (elevation of anti-TPO, hypoechoic thyroid) were present prior to RIT. Noteworthy, the TSI level was in the upper range of normal.[br][bold]Conclusion[/bold]: Taking into consideration the reported risk factors, we can better identify the patients who are more prone to develop post radioactive iodine Graves[apos] disease prompting a closer follow-up. Therefore, initial evaluation including measurements of autoimmune thyroid antibodies, thyroid scintigraphy, and sonography prior to RIT should be strongly considered.[br][br]Nothing to Disclose: SM, DD, PL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 829 192 1467 SUN-440 PO56-02 Sunday 1264 2012


1261 ENDO12L_SUN-441 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) An Anterior Mediastinal Mass: Thymic Hyperplasia Associated with Hyperthyroidism Margaret Gladysz, Joe Chehade University of Florida, Jacksonville, FL [bold]Clinical Cases[/bold][br][bold]Patient 1:[/bold] A 25-year-old woman presented with an anterior chest pain, associated with shortness of breath, intermittent palpitations, and heat intolerance.[br]Physical examination was notable for an anxious appearing woman with irregularly irregular tachycardia at the rate of 108bpm and brisk tendon reflexes. Thyroid gland was diffusely enlarged, with estimated weight of 45 grams. Findings on the remainder of the examination were unremarkable.[br]Laboratory evaluation revealed, suppressed TSH at [lt]0.01mIU/mL (0.34-4.82mIU/mL), elevated FT3 at 7.2pg/mL (2.3-4.2pg/mL) and TT4 at 17.3mcq/dL (4.5-12.1mcq/dL). Chest CT (computed tomography) demonstrated an anterior mediastinal soft tissue mass in the expected region of thymus, not seen on previous CT scan a year earlier.[br][bold]Patient 2:[/bold] A 51-year-old woman presented with complaint of a progressive, substernal chest pain associated with episodes of nausea and shortness of breath.[br]Physical examination was unremarkable, except for sinus tachycardia with 115bpm. Her thyroid work up revealed suppressed TSH at 0.19mIU/mL and elevated TT4 at 13.8mcq/dL. CT angiogram of the chest revealed an abnormal mass-like, soft tissue density in the anterior mediastinum measuring up to 3.4cm x1.6cm with an adjacent nodular density measuring 2.2cm x 1.6cm.[br]Both women underwent core biopsy of the mediastinal mass. Pathology was consistent with thymic tissue hyperplasia. After treatment with anti-thyroid medications for 6 to 12 months, follow up CT angiogram demonstrated marked regression of thymus mass.[br][bold]Clinical Lesson(s)/Conclusion[/bold][br]Lymphoid thymus hyperplasia is a known and benign association of Graves[apos] disease. TR-Ab (thyroid hormone receptor antibody) and expression of the thyrotropin receptor in the thymus likely play a principal role in the development of hyperplasia.[br]The assessment of thymus hyperplasia is best done with CT and/or MR (magnetic resonance) with chemical shift imaging. Both showing homogeneously decreased in signal intensity on T2 images. Furthermore, FDG PET scan was found to be helpful in demonstrating a CSR (lower chemical shift ratio) with hyperplasia. The regression of the thymic mass and the decrease of soft tissue attenuation may represent reduction in hyperplastic thymic tissue. Recognizing the association between thymic hyperplasia and hyperthyroidism may prevent unnecessary invasive diagnostic studies and surgical procedures.[br][br]Nothing to Disclose: MG, JC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 890 192 1468 SUN-441 PO56-02 Sunday 1265 2012


1262 ENDO12L_SUN-442 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Severe Thyroid-Associated Ophthalmopathy in a Euthyroid AIDS Patient Ashutosh Pareek, Maria Keatts Chavez, Agustin Busta Beth Israel Medical Center, New York, NY [bold]Objective[/bold]: To present a case of severe thyroid associated ophthalmopathy (TAO) in a euthyroid AIDS patient.[br][bold]Case[/bold]: A 43-year-old African-American man with acquired immunodeficiency syndrome(AIDS), hypertension, schizophrenia, deafness and blindness due to cryptococcal meningitis was referred to endocrine in late 2011 for proptosis evaluation. Physical exam: TAO type II- bilateral severe proptosis, conjunctival edema and erythema. Rest of the physical exam, including the thyroid exam, was unremarkable. Labs: TSH 2.29mU/L (0.55-4.78), free T3 2.0pg/ml (2.3-4.2), free T4 0.7ng/dL (0.7-1.7), anti TPO Ab [lt]10IU/ml (0-34.9), and thyroid stimulating immunoglobulin (TSI) of 345 (normal[lt]140). CT of the orbits without contrast two months prior to consult reported bilateral proptosis, no abnormal soft tissue or mass effect in the retrobulbar fat or orbital apices bilaterally. Cavernous sinus was normal. The patient[apos]s condition since has remained stable.[br][bold]Discussion[/bold]: This is a case of an AIDS patient with severe Graves[apos] ophthalmopathy (GO)in the setting of normal thyroid function tests (TFTs) and is reported in 5-10% of such patients[sup]1[/sup]. TSI shows significant association with the clinical features of TAO[sup]4[/sup]. TSH receptor antibody (TSHr ab) is also implicated in GO pathogenesis[sup]2[/sup]. This antibody, directed against thyroid follicular cells, recognizes antigenic epitopes that are shared by tissues contained within the orbital space[sup]2[/sup].[br]Our patient was being treated with protease inhibitors (PI), which may have contributed to GO by remodeling adipocytes in the setting of autoimmune thyroid disease[sup]3[/sup]. PIs are a well known cause of lipodystrophy and changes in adipogenesis and adipose cell turnover have the potential to disrupt adipose cell form and function leading to accumulation of fat in the retro-orbital area[sup]3[/sup].[br][bold]Conclusion[/bold]: This case is unique in that an AIDS patient, being treated with a PI, suffered from GO in the setting of normal thyroid function tests. Possible fat tissue proliferation by PIs likely contributed to the GO. Scarce data exists about the factors leading to GO in AIDS patients who are on a PI. We believe further investigation to better grasp the pathoimmunology of this entity will be a significant contribution in the treatment of euthyroid Graves[apos] ophthalmopathy.[br][br](1) Mallika PS. Thyroid associated ophthalmopathy- a review. Malaysian Family Physician. 2009; Vol.4, No.1. (2) K. Kazuo, T. Fujikado, G. Ohmi, J. Hosohata, and Y. Tano. Value of Thyroid Stimulating Antibody in the Diagnosis of Thyroid Associated Ophthalmopathy of Euthyroid Patients. Br J Ophthalmol. 1997 December; 81(12): 1080[ndash]1083. (3) Sorisky A, Gagnon AM. Clinical Implications for Adipose Tissue Remodelling: Adipogenesis and Apoptosis. Can Journal of Diabetes. 2002; 26 (3): 232-240. (4) Ponto KA, Kanitz M, Olivo PD, Pitz S, Pfeiffer N, Kahaly GJ. Clinical relevance of thyroid-stimulating immunoglobulins in graves[apos] ophthalmopathy. Ophthalmology. Nov 2011;118(11):2279-85.[br][br]Nothing to Disclose: AP, MKC, AB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1829 192 1469 SUN-442 PO56-02 Sunday 1266 2012


1263 ENDO12L_SUN-443 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Pseudotumor Cerebri in a Patient with Graves Disease, Is There an Association? Sonia M Gibson, Abdul Abou-Samra Wayne State University, Detroit, MI [bold]Backround[/bold]: Idiopathic Intracranial Hypertension (IIH), commonly referred as pseudotumor cerebri, is characterized by a clinical picture dominated by signs and symptoms of intracranial hypertension with normal composition of cerebrospinal fluid (CSF) and neuro-imaging that shows no other cause of IHH. Several endocrinopathies have been linked to IHH, including Addison[apos]s disease and hypoparathyroidism.[br]We present a case of a 25 year old African-American woman who presented in May 2011 complaining of severe headaches. Her headaches began in February 2010 after a C-section. Past medical history revealed an emergency-room (ER) visit in January 2011 with intractable vomiting and visual disturbances. She had papilledema and a lumbar puncture (LP) with an opening pressure of 48 mm/Hg indicating high intracranial pressure (ICP) and normal CSF composition. Head computed tomography (CT) showed no intracranial process. The patient was not taking oral contraceptive pills or any other medication. Acetazolamide was started and a follow-up visit was arranged. In June 2011, the patient visited the ER again with similar complaints; LP offered little symptomatic relief. Persistent sinus tachycardia was noted. Thyroid function tests (TFTs) revealed TSH [lt]0.008 uIU/ml, free T4 of 3.2 ng/dL and total T3 of 355 ng/dL. Patient had thyromegaly, mild periorbital edema, no exophthalmos or lid lag. Methimazole 20 mg daily was started. The thyroid uptake was 51% at 24 hrs and the scan was homogenous; compatible with Graves[apos] disease. A few weeks post discharge, her headaches improved; however, the patient stopped Methimazole in August 2011 due to GI side effects. In September 2011 she presented with acute right eye blindness secondary to worsening ICP causing optic nerve damage. She was thyrotoxic and a head CT showed increased size of ventricles. A ventriculo-peritoneal (VP) shunt was placed to alleviate the IC pressure. The patient also underwent ablation with 14.9 mCi of [sup]131[/sup]I-Na in December 2011. Symptoms cleared as her TFTs approached a euthyroid state. She remained legally blind in her right eye.[br][bold]Conclusion[/bold]: IIH is a rare entity in which endocrine pathologies have been previously described as possible etiologies. Graves[apos] Disease may be associated with IIH, with few cases previously reported in the literature. This seems to be the first case reported of IIH causing blindness associated with Graves[apos] disease.[br][br]Nothing to Disclose: SMG, AA-S 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2001 192 1470 SUN-443 PO56-02 Sunday 1267 2012


1264 ENDO12L_SUN-444 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) A Case of Autoimmune Lymphocytic Thyroiditis Causing Thyroid Storm Sara Ahmadi, LaDale K St Clair University of Texas Medical Branch, Galveston, TX [bold]Introduction: [/bold]Thyroid storm is uncommon but serious complication of thyrotoxicosis and is usually associated with Graves[apos] disease. It involves decompensation of one or more organs and it has high mortality. It is usually triggered by precipitating factor like infection or surgery. We are presenting a case of Autoimmune Lymphocytic thyroiditis causing thyroid storm.[br][bold]Clinical case: [/bold]A previously healthy 27 year old male was admitted in our medical ward due to a three day history of fever, nausea/vomiting and palpitation. Initial physical exam revealed a weak, anxious, agitated male. Vital signs: Temp 101[deg] F, Pulse 113 bpm, BP 154/96. He had no Graves[apos] ophthalmopathy, no significant thyromegaly and no bruit audible over the thyroid. He had hyperreflexia, hand tremor and very warm hands. Initial laboratory work up revealed undetectable TSH, elevated free T4 [gt]7 ng/dL, normal CBC, normal electrolytes and elevated liver enzymes. Per Burch and Wartofsky[apos]s criteria to diagnose thyroid storm, our patient had a total score of 45 points (temp 15, agitation 10, nausea/vomiting 10, pulse 10). Patient was started on medical management with methimazole (MMI), propranolol, iodine solution and hydrocortisone. Iodine solution was stopped after 5 days which caused worsening of thyroid and liver function. Thyroid and Liver function both improved after patient was restarted on Iodine. Despite receiving high dose MMI and Iodine solution for more than two weeks patient continued to be hyperthyroid. ENT consult was placed for possible thyroidectomy due to requiring high does MMI (120 mg daily) and not being able to monitor the patient closely as outpatient due to his social issues. Thyroidectomy was done after patient was made clinically stable. Further lab results revealed elevated TPO Abs of 2570 WHO units [Normal 0-100], TRAB 26.29 IU/L [Normal [lt]1.75] and negative TSI 116% [Normal [lt]129]. Pathology report revealed lymphocytic thyroiditis and no evidence of Graves[apos] disease. Patient remained clinically stable during the remainder of his hospitalization and clinic follow up.[br][bold]Conclusion:[/bold] It is important to determine the etiology of thyrotoxicosis because treatment and outcome vary. Despite our extensive search we just found two cases of sub acute thyroiditis and no cases of Lymphocytic thyroiditis reported causing thyroid storm. These cases demonstrate that we should consider thyroiditis and Lymphocytic thyroiditis as part of the differential diagnosis in thyroid storm.[br][br]Nothing to Disclose: SA, LKSC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1155 192 1471 SUN-444 PO56-02 Sunday 1268 2012


1265 ENDO12L_SUN-445 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Pheochromocytoma Presenting as Recurrent Goiter in Patient with Graves Disease and Neurofibromatosis Urseline Altovise[apos] Hawkins, Shema R Ahmad University of Mississippi Medical Center, Jackson, MS Background: The link between paroxysmal thyroid swelling, and pheochromocytoma has been well described in the past; however, it remains a scarcely reported entity. Here, we present a patient with Graves disease, and recurrent goiter, as clinical manifestations of pheochromocytoma.[br]Clinical Case: A 40 yo AAM with history of neurofibromatosis was admitted to the hospital for planned excision of right thoracic pseudomeningocele. Endocrinology was consulted after work-up of post-operative tachycardia revealed thyrotoxicosis. Pt reported several month history of unintentional weight loss, diarrhea, anxiety, tachycardia, and tremor. Physical exam was notable for orbitopathy, goiter, and thyroid bruit. TSH 0.016 uIU/mL (normal range 0.27-4.20), Free T4 [gt]6.50 ng/dL (0.93-1.70), TSI 4.6 ([lt]1.3), Thyrotropin Receptor Antibody [gt]40.00 IU/L (0.00-1.75). Pt was diagnosed with Graves disease, and started on Methimazole 20mg BID, and Propranolol. His first outpatient follow-up revealed marked improvement in his goiter, with no thyroid bruit on exam, as well as resolution of the tachycardia. Pt presented back several months later, and reported sudden increase in size of goiter, with new onset compressive symptoms. He reported compliance with Methimazole 20mg BID. A large goiter, with return of thyroid bruit was noted on physical exam. Thyroid US revealed thyromegaly with numerous hyper, and hypoechoic nodules, mostly without increased blood flow suggesting edema. Plasma normetanephrine 468 pg/mL ([lt]148), plasma metanephrine 64 pg/mL ([lt]57), total metanephrine 532 pg/mL ([lt]205). Further localization work-up for pheochromocytoma remains pending at this time.[br]Conclusion: Pheochromocytoma has been linked to numerous clinical findings, including paroxysmal thyroid swelling. Cases of pheochromocytoma demonstrating such swelling, including ours, are norepinephrine-secreting. In our case, we were prompted to evaluate for pheochromocytoma, in a patient with Graves disease, and recurrent goiter. We present this case for illustration, as we feel that others should be mindful of this association.[br][br]Nothing to Disclose: UAH, SRA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 550 192 1472 SUN-445 PO56-02 Sunday 1269 2012


1266 ENDO12L_SUN-446 POSTER SESSION: Autoimmune Thyroid Disease Case Reports (1:30 PM-3:30 PM) Autoimmune Polyglandular Syndrome: An Unusual Presentation with Empty Sella, Premature Ovarian Failure, and Hashimoto Thyroiditis Associated with Thyroid Cancer Maria Carolina Rios, Juliana Pereira Passaglia, Carolina GS Leaes, Julia FS Pereira-Lima, Miriam da Costa Oliveira ISCMPA/Universidade Federal de Ci[ecirc]ncias da Sa[uacute]de de Porto Alegre (UFCSPA), Porto Alegre, Brazil Introduction: Autoimmune polyglandular syndromes are rare disorders characterized by failure of several endocrine glands, as well as non-endocrine organs, associated with immunemediated tissue destruction. Clinical Case: We report a case of a 38-year-old female patient presented with secondary amenorrhea. She reported menarche at 12 years, followed by irregular cycles, single delivery without complications at 21, and amenorrhea at age 26. At that time, she was diagnosed with primary hypothyroidism and levothyroxine therapy was started. With 37 years, she underwent total thyroidectomy, after detection of nodules. Multicentric papillary carcinoma was diagnosed, with cervical lymph node metastasis. Positive anti-thyroperoxidase antibodies (442 U/mL) and negative anti-thyroglobulin antibodies were detected. Complementary radioiodine therapy was performed. During follow-up, the patient reported headache and dizziness, and magnetic resonance imaging (MRI) of the brain was performed, showing an empty sella and the patient was then referred to the neuroendocrinology service. The patient denied other comorbidities and use of medications other than levothyroxine (150 mcg/day). She had a positive family history for thyroid cancer and type 2 diabetes mellitus and negative for early menopause or autoimmune diseases. Physical and neurological examination revealed no other abnormalities. Biochemical tests revealed impaired glucose tolerance and mixed dyslipidemia. Hypergonadotropic hypogonadism was confirmed. Baseline cortisol, cortisol after ACTH stimulation, prolactin, and IGF-1 levels were normal. Antipituitary antibodies (APA) were negative and anti-21-hydroxylase antibodies (21OHAb) were positive (6.7 U/mL; reference value [lt] 1). The patient had osteopenia of the lumbar spine (T score - 1.73). In the last examination, serum thyroglobulin and whole body screening showed no evidence of recurrence and/or residual tumor. The patient is currently on therapy with calcium carbonate, vitamin D, and simvastatin and on levothyroxine, estrogen, and progestagen replacement therapy. Conclusion: It[apos]s a rare case of polyglandular syndrome type II in a patient who presented with premature ovarian failure, Hashimoto[apos]s thyroiditis, and empty sella associated with a diagnosis of differentiated thyroid carcinoma. This case probably represents the first report on this tumor in a patient with polyglandular disorder.[br][br]Nothing to Disclose: MCR, JPP, CGSL, JFSP-L, MdCO 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 429 192 1473 SUN-446 PO56-02 Sunday 1270 2012


1267 ENDO12L_SUN-449 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Glucocorticoid-Remediable Aldosteronism and Adrenal Adenoma: A Diagnostic Challenge Resolved with Adrenal Vein Sampling Lindsay B Harrison, Richard J Auchus, Michael J McPhaul University of Texas at Southwestern, Dallas, TX; University of Michigan, Ann Arbor, MI [bold]Introduction:[/bold] Glucocorticoid-Remediable Aldosteronism (GRA) is a rare cause of primary aldosteronism (PA) but should be included in the differential diagnosis for young patients.[br][bold]Clinical Case:[/bold] A 39-year-old Hispanic man was admitted for hypokalemia (2.4 mEq/L, nl 3.6-5.0). He was diagnosed with hypertension at age 28, which was uncontrolled with lisinopril, and he had also required potassium supplementation in the past. Family history included mother with resistant hypertension and a brother with hypertension diagnosed at age 24. The serum aldosterone was 47 ng/dl at 1100 and 10.4 ng/dl at 2300 (n [lt]31) with a plasma renin activity (PRA) of [lt]0.6 ng/mL/hr. An intravenous sodium infusion revealed a basal 0800 aldosterone of 92 ng/dl and a 4-hour post-load aldosterone of 20 ng/dl (both with PRA [lt]0.6ng/ml/hr), confirming PA. An abdominal CT scan revealed a left 2.1x2.1 cm 6.3 Houndsfield units (HU) adrenal adenoma. Given his young age and [gt]1 cm adenoma with a normal contralateral adrenal gland, a presumptive diagnosis of aldosterone-producing adenoma and a plan of surgical resection were considered. Nevertheless, adrenal vein sampling (AVS) with cosyntropin infusion (50 mcg/h) was performed to confirm the diagnosis; however, AVS showed markedly elevated aldosterone at all sites without lateralization:[br]Right AV 9,220 ng/dl (cortisol 540 [mu]g/dl), A/C ratio 17.1[br]Left AV 22,520 ng/dl (cortisol 1,248 [mu]g/dl), A/C ratio 18.0[br]IVC 118 ng/dl (cortisol 37 [mu]g/dl), A/C ratio 3.2[br]Peripheral vein 290 ng/dl (cortisol 37 [mu]g/dl), A/C ratio 7.8[br]This dramatically elevated and virtually identical aldosterone from both adrenals under cosyntropin stimulation and the diurnal rhythm of aldosterone suggested ACTH-dependent aldosterone secretion. A suppression test was performed with dexamethasone 0.5 mg every 6 hours for 2 days, after which the 0800 serum aldosterone [lt]1.6 ng/dl, consistent with GRA (familial hyperaldosteronism type 1). Genetic testing to confirm the diagnosis is underway. Rather than surgery, the patient is managed with amiloride and has normal BP and improved potassium.[br][bold]Clinical Lessons:[/bold] Many sources advocate sending young patients with PA and a [gt]1 cm adenoma in one adrenal gland with a normal contralateral adrenal gland directly to surgery without confirmatory AVS. To our knowledge, this is the first patient with GRA to undergo AVS, and illustrates the need to consider that genetic causes of aldosterone excess should be excluded prior to surgical intervention.[br][br]Disclosures: RJA: Investigator, Jansen Pharmaceuticals. MJM: Investigator, Corcept Therapeutics, Novartis Pharmaceuticals. Nothing to Disclose: LBH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1590 193 1474 SUN-449 PO45-02 Sunday 1271 2012


1268 ENDO12L_SUN-450 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Aldosterone and Cortisol Co-Secreting Adenomas: A Retrospective Study Andreas G Moraitis, Mitra Lynn Rauschecker, Smta Baid Abraham, Constantine A Stratakis National Institutes of Health, Bethesda, MD [bold]Background[/bold]: In 1977, the first case of a single adenoma that was producing aldosterone along with cortisol was reported[i]. Subsequently, co-secretion of cortisol in cases of primary hyperaldosteronism (PA) has been reported in several case reports and small series with an apparent prevalence up to 21%[ii, iii].[br][bold]Patients and methods[/bold]: We retrospectively studied 60 consecutive patients with primary hyperaldosteronism (PA) referred and treated at the NIH. The diagnosis of PA was confirmed by normal saline suppression test and/or oral salt loading test. All patients underwent adrenal CT scan and adrenal venous sampling. Criteria for AVS lateralization: aldosterone/cortisol ratio [gt]4:1 and contralateral gland suppression to periphery. Inpatient evaluation for autonomous cortisol secretion included 24hr urine free cortisol (UFC), 2 day low dose dexamethasone suppression test (LDDST) and 2 day high dose dexamethasone suppression test (HDDST), with 24hr 17-OH-corticosteroids (17OHS) and serum cortisol on day 2, diurnal cortisol and ACTH measurements.[br][bold]Results: [/bold]Among 60 consecutive patients referred to the NIH for evaluation and management of primary aldosteronism, 8 patients (13%) were found to have biochemical evidence of autonomous cortisol secretion. Using a cutoff for serum cortisol of 1.8mcg/dl on day 2 of the LDDST and HDDST, the LDDST showed no suppression in all, 7 patients tested; HDDST also showed no suppression in all 6 patients tested. 24hrs -17OHS on day 2 of LDDST were more than 4.4mg in all 7 patients tested. In contrast, 7 out of 8 patients (87%) had normal UFCs, while only one patient with a 2.5cm single adenoma had elevated UFCs (four times the upper normal reference range).[br][bold]Conclusions[/bold]: In our series, the prevalence of cortisol co-secretion in patients with PA was 13%, underlying the importance of screening for cortisol co-secretion in patients with PA. The 2 day LDDST and HDDST were equally sensitive (100%). 24hrs 17OHS on the second day of both the LDDST and HDDST were 100% sensitive using cut-off values of 4.4 mcg/day and 3.7mcg/day respectively. In addition, a decrease of the 17OHS by less than 50% during the LDDST and HDDST was also 100% sensitive for autonomous cortisol secretion.[br][br][i] Hogan MJ, Schambelan M, Biglieri EG (1977)Concurrent hypercortisolism and hyper-mineralocorticoidism. Am J Med 62: 777-782. [ii] Adachi J, Hirai Y, Terui K, Nakano T, Fukuda Y, Suda T [amp] Sasano H. A report of 7 cases of adrenal tumors secreting both cortisol and aldosterone. Internal Medicine 2003 42 714[ndash]718. [iii] Piaditis GP, Kaltsas GA, Androulakis II, Gouli A, Makras P, Papadogias D, Dimitriou K, Ragkou D, Markou A, Vamvakidis K, Zografos G, Chrousos G (2009) High prevalence of autonomous cortisol and aldosterone secretion from adrenal adenomas. Clin Endocrinol (Oxf) 71: 772-778.[br][br]Sources of Research Support: Eunice Shriver Kennedy National Institute of Child Health and Human Development, National Institutes of Health (NIH), Bethesda, MD.[br][br]Nothing to Disclose: AGM, MLR, SBA, CAS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1814 193 1475 SUN-450 PO45-02 Sunday 1272 2012


1269 ENDO12L_SUN-451 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Testosterone-Secreting Adrenal Adenoma in a Woman with Non-Classical Congenital Adrenal Hyperplasia Smita Kargutkar, Arthur Chernoff Albert Einstein Healthcare Network, Philadelphia, PA [bold][underline]INTRODUCTION:[/underline][/bold] We report a case of a woman with delayed onset congenital adrenal hyperplasia with testosterone-secreting adrenal adenoma.[br][bold][underline]CASE REPORT:[/underline] [/bold]A 33 year woman presented with amenorrhea, 20 lbs weight gain and new hirsutism.[br][bold][underline]PMH:[/underline][/bold] Menarche at age 12, regular menses, G2P2A0 no problems conceiving.[br][bold][underline]PE:[/underline][/bold] Acanthosis nigricans, Ferriman Gallwey Score 12.[br][underline][bold]Labs:[/bold][/underline] Estradiol 71 pg/mL (10-200), FSH 2.9 mIU/mL (2.5-10.2), LH 2.9 mIU/mL (1.9-12.5), Prolactin 7.4 ng/mL (3-30), TSH 1.91 mIU/L (0.40-5.50), 17-[alpha]-OH-progesterone 360 ng/dL ([lt]185), DHEA-S 76 mcg/dl (40-325), Testosterone Total 236 ng/dl (2-45), Free 27.6 pg/ml (0.1-6.4), Androstenedione 301 ng/dL (65-270), Fasting insulin level 20 micro IU/mL ([lt]17), Glucose 88 mg/dL (65-99).[br][underline][bold]Cosyntropin Stimulation Test (250 mcg):[/bold][/underline] At 0, 30 and 60 minutes respectively: 17-[alpha]-OH-progesterone (43 ng/dl, 475 ng/dl, 514 ng/dl) (10-120) Androstenedione 345 ng/dl, 552 ng/dl, 447 ng/dl (47-268) Cortisol 6.6mcg/dL, 10.6 mcg/dL, 10.2mcg/dL) (4-22). Dexamethasone 0.5mg HS and Metformin 500mg BID was started but hirsutism and acne got worse. After dexamethasone 0.5mg Q6 H for 48 hours, the 24 hour urine cortisol was 18 mcg, not suppressed.[br]Abdomen and pelvis CT showed a 2.2 cm enhancing right adrenal mass and U/S of pelvis showed 2.3 x 2.1 x 2.0 cm hyperechoic mass in right ovary characteristic of a dermoid tumor. Adrenal and ovarian vein sampling failed due to technical problems.[br]A right adrenalectomy yielded pathology consistent with an adrenal adenoma. Post-op testosterone level normalized to total 25ng/dl (2-45) and free 0.6pg/mL (0.1-6.4). However, 17-[alpha]-OH progesterone remained elevated to 360ng/dl ([lt]185). Dexamethasone 0.5 mg q hs was restarted which decreased the 17-[alpha]-OH progesterone to 70 ng/dL. She started having regular menstrual cycles post-op and Ferriman Gallwey Score decreased to 3.[br][bold][underline]DISCUSSION:[/underline][/bold] A functioning adrenal tumor was discovered in a patient with non-classical congenital adrenal hyperplasia due to (presumed) 21-hydroxylase deficiency. The presence of this defect in steroid hormonogenesis appears to be responsible for transforming a (non-suppressible) cortisol producing adenoma into a testosterone secreting adenoma.[br][bold][underline]CONCLUSION:[/underline][/bold] Adrenal adenomas can occur in the setting of non-classic adrenal hyperplasia from continuous adrenal stimulation by elevated ACTH secondary to loss of negative feedback effect. Testosterone-secreting adrenal adenomas could be a manifestation of underlying defects in hormonogenesis.[br][br]1.Selma Feldman Witchel1 and Ricardo Azziz2,3 Nonclassic Congenital Adrenal Hyperplasia; International Journal of Pediatric Endocrinology; Volume 2010, Article ID 625105,. 2. Shimshi M, Ross F, Goodman A, Gabrilove JL, Virilizing adrenocortical tumor superimposed on congenital adrenocortical hyperplasia; Am J Med. 1992 Sep;93(3):338-42. 3. A. P. Van Seters, W. Van Aalderen, A. J. Moolenaar,M. C. B. Gorsiro, F. Van Roon AND E. T. Backer; Adrenocortical Tumour in Untreated Congenital Adrenocortical Hyperplasia Associated with Inadequate ACTH Suppressibility; Clinical Endocrinology (1981) 14,325-334. 4.Pekka Leinonen; Taplo Ranta; Rita Siegberg; Risto Pelkonen; Piilvl Helkkllii; and Arvi Kahri;Testosterone-secreting virilizing adrenal adenoma with human chorionic gonadotrophin receptors a.nd 21-hydroxylase deficiency; Clinical Endocrinology (1991) 34. 31-35.[br][br]Nothing to Disclose: SK, AC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 425 193 1476 SUN-451 PO45-02 Sunday 1273 2012


1270 ENDO12L_SUN-452 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Is Primary Hyperaldosteronism Due to Unilateral Hyperplasia Actually Early Evidence of Bilateral Hyperplasia? Mitra Rauschecker, Andreas Moraitis, Constantine Stratakis, Smita Abraham National Institutes of Health (NIH), Bethesda, MD Background: Primary hyperaldosteronism (PA) due to unilateral disease is typically associated with an aldosterone producing adenoma (APA) and rarely to unilateral hyperplasia. We describe three cases of PA in which imaging and adrenal venous sampling (AVS) were consistent with APA, however pathology revealed adrenal hyperplasia. Criteria for AVS lateralization: aldosterone/cortisol ratio [gt]4:1 and contralateral gland suppression to periphery.[br]Case 1: 42 y old woman with PA; CT scan demonstrated a 1 cm left adrenal nodule, although MRI demonstrated bilateral thickening. AVS demonstrated lateralization to the left side; patient underwent left adrenalectomy. Pathology revealed adrenal hyperplasia, without evidence of a nodule. On post-operative day 1, aldosterone was undetectable, blood pressure (BP) was improved, with reduced dosages of anti-hypertensive medications, and hypokalemia resolved. One month post-operatively, aldosterone and renin were undetectable. She will follow-up in 6 months. Case 2: 55 y old man with PA; AVS demonstrated lateralization to the right side. Adrenal CT showed a 2 cm nodule on the right gland. Patient underwent right adrenalectomy. Pathology revealed adrenal hyperplasia, without evidence of a nodule. On post-operative day 1, aldosterone was 9.5 ([lt]21 ng/dl), with an undetectable renin level. BP was controlled on the same anti-hypertensive regimen and hypokalemia resolved; however, 1 month after surgery, patient had recurrence of original symptoms of fatigue and muscle weakness. Six months after surgery, repeat aldosterone/renin ratio was 15.3 and saline suppression test showed an aldosterone level of 6.6 ng/dl at 4 hours. Case 3: 62 y old man with PA; CT scan revealed nodularity of both adrenal glands with a 0.8 cm nodule on the right. AVS demonstrated a dominant right adrenal (ratio 12:1), however, contralateral gland suppression was questionable. Pathology revealed diffuse hyperplasia without a nodule. Post-operatively, aldosterone was undetectable, hypokalemia resolved, and BP was mildly improved on the same dosage of anti-hypertensive medication. Patient was lost to follow-up.[br]Conclusion: We hypothesize that unilateral hyperplasia may be an early presentation of bilateral hyperplasia in patients with PA. Our case series shows a clinical spectrum of disease with each patient representing a different stage. In patients with PA due to unilateral hyperplasia, follow-up for recurrence of disease may be warranted.[br][br]Nothing to Disclose: MR, AM, CS, SA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 545 193 1477 SUN-452 PO45-02 Sunday 1274 2012


1271 ENDO12L_SUN-453 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Benign Adrenal Adenoma Co-Secreting Excess Mineralocorticoids and Glucocorticoids Hans K Ghayee, Vivienne Yoon, Hironobu Sasano, Stacey Woodruff, Kelley S Carrick, Richard J Auchus University of Texas Southwestern Medical Center, Dallas, TX; Tohoku University School of Medicine and Hospital at Sendai, Sendai, Japan; University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX; University of Michigan, Ann Arbor, MI Primary aldosteronism causes hypertension and hypokalemia. However, secretion of other mineralocorticoids in the zona fasciculata including 11-deoxycorticosterone (DOC) and corticosterone, can lead to hypertension and hypokalemia. An adrenal tumor producing 17-deoxysteroids in addition to aldosterone might cause an underestimation of mineralocorticoid excess in a patient with a benign adrenal mass. A 52-year old man presented with a history of uncontrolled hypertension, hypokalemia, and type 2 diabetes since the age of 36 was found to have right adrenal nodule during work-up for hypertension and hypokalemia. The patient did not complain of skin thinning or easy bruising. He was taking four agents for his blood pressure including spironolactone and potassium supplements. The serum aldosterone was 11ng/dL with a renin 0.25ng/mL/hr, K= 3.4mmol/L, while taking lisinopril, suggestive of mild primary hyperaldosteronism. The early morning plasma ACTH was 16pg/mL with a serum cortisol of 16.1[micro]g/dL and suppressed DHEA-S; the urinary free cortisol was normal. The serum cortisol fell only to 3.9[micro]g/dL during a 1mg dexamethasone suppression test and midnight salivary cortisols were abnormal at:.110,.140, and.220ng/dL ([lt]0.090), consistent with mild ACTH-independent Cushing Syndrome. An abdominal CT scan showed growth of the right adrenal nodule (18U) from 3.5cm x 3.5cm to 4.6cm x 4.0cm over a 2 year period, with a stable 1cm left adrenal nodule. Due to the enlarging mass and evidence of autonomous cortisol production, right adrenalectomy was performed. An adrenal adenoma without evidence of malignant features was confirmed histopathologically. Post-operatively, AM cortisol was 1.0[micro]g/dL. After the procedure, he no longer required spironolactone. Immunohistochemistry (IHC) from the tumor tissue is being analyzed for steroidogenic enzyme expression. This is a case of combined glucocorticoid and mineralocorticoid excess with an enlarging adrenal tumor. The degree of hypertension and hypokalemia was disproportionately severe for his degree of aldosterone excess. IHC is being performed to potentially determine the contribution of DOC and corticosterone[apos]s effects in causing hypertension and hypokalemia in addition to aldosterone and cortisol. This case underscores the difficulties in evaluating the hormone production of adrenal tumors without comprehensive steroid profiling.[br][br]Nothing to Disclose: HKG, VY, HS, SW, KSC, RJA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1819 193 1478 SUN-453 PO45-02 Sunday 1275 2012


1272 ENDO12L_SUN-454 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) A Case of Surgically Uncurable Aldosterone-Producing Adenoma [mdash] Reconsideration for AVS Criteria and Surgical Indication in Such Unusual Cases Isao Kurihara, Hirotaka Shibata, Kazutoshi Miyashita, Ayano Murai-Takeda, Yuko Mitsuishi, Rie Jo, Takako Ohyama, Akira Miyajima, Mototsugu Ohya, Kaori Kameyama, Kuniaki Mukai, Hiroshi Itoh School of Medicine, Keio University, Tokyo, Japan; School of Medicine, Keio University, Tokyo, Japan; School of Medicine, Keio University, Tokyo, Japan; School of Medicine, Keio University, Tokyo, Japan [Introduction] In the subtype classification of primary aldosteronism (PA), adrenal vein sampling (AVS) is the standard test to differentiate unilateral from bilateral causes of PA. Although there is a controversy whether which criteria is most accurate, most clinicians use [quot]lateralized ratio: high side-to-low side ratio of aldosterone/cortisol (A/C) ratio [gt]4[quot] and/or [quot]contralateral ratio: low side-to-inferior vena cava ratio of A/C ratio [lt]1[quot] for diagnosis of unilateral hypersecretion of aldosterone. We recently developed a novel diagnostic approach for APA, using a specific CYP11B2 immunohistochemistry on resected tissues, and we employed it to validate our diagnosis.[br][Case presentation] A 56-year old man visited a primary care physician because hypertension was pointed out at annual health checkup. He was refered to our hospital for confirmation of PA because he had hypokalemia and aldosterone-renin ratio was positive. We confirmed this case as PA by oral salt loading test (urine aldosterone (36.8mcg/day), and went to the step of subtype classification. Adrenal CT scan revealed bilateral nodular lesions and we underwent AVS for judging surgical indication. Results in post ACTH-stimulated AVS were following;[br]Right adrenal vein A[pg/ml]/C[mcg/dl]=51900/1035.6=50.1[br]Left adrenal vein A/C=18700/947.1=19.7[br]Inferior vena cava A/C=728/21.5=33.9[br]Although the lateralized ratio was 2.5 which was less than our cut-off, the contralateral ratio was suppressed to 0.6 and urine aldosterone excretion was markedly high, we decided to undergo right adrenalectomy. After right adrenalectomy, hypokalemia was cured, but hypertension persisted and aldosterone-renin ratio failed to get normalized. Microscopic examination on resected tissue revealed multiple nodules with a variable size. Only a single nodule showed positive for both CYP11B2 and HSD3B2, but negative immunostainings for CYP17, demonstrating that the nodule was compatible with APA. We speculated that unusual postoperative hypersecretion of aldosterone can be attributed to another APA in opposite side of adrenal.[br][Conclusion] The CYP11B2 immunostainings on resected adrenal tissue of PA is a powerful approach to distinguish from which nodule aldosterone is secreted. The present case demonstrate that bilateral APAs cannot be ruled out in the case of confirmed PA with the lateralized ratio between 2 and 4 and suppressed contralateral ratio.[br][br]Nothing to Disclose: IK, HS, KM, AM-T, YM, RJ, TO, AM, MO, KK, KM, HI 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1394 193 1479 SUN-454 PO45-02 Sunday 1276 2012


1273 ENDO12L_SUN-455 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Adrenal Amyloidosis with Co-Existing Primary Hyperaldosteronism Mitra Rauschecker, Robert Shamburek, James Balow, Smita Abraham National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD Background: Hereditary amyloidosis is a rare disease characterized by deposition of amyloid fibrils in multiple organs, including the kidneys, liver, heart, and the adrenal glands. It is rarely reported in conjunction with disorders of the adrenal gland.[br]Clinical Case: 60 y old Hispanic male with a 10 year history of type II diabetes mellitus, non-dialysis dependent renal failure (eGFR: 18 ml/min/1.73 sq. m) with rapid onset nephrotic level proteinuria and primary aldosteronism (PA) was evaluated for optimal management of PA. Aldosterone was 35 ng/dl ([lt]21), and plasma renin was suppressed at [lt]0.6 ng/ml/hr on presentation. Adrenal venous sampling revealed unilateral disease of the left gland. Patient opted for surgery as previous management with anti-hypertensives, including aldosterone antagonists, was unsuccessful. Post-operatively, blood pressure improved: two anti-hypertensives were reduced in dosage, while the third was discontinued. Hypokalemia resolved. Plasma renin was detectable at 2.7 ng/ml/h and aldosterone was suppressed at [lt]4.0 ng/dl on post-operative day 1. Adrenal pathology revealed presence of abundant amyloid without evidence of a nodule or hyperplasia. Kidney biopsy also showed amyloid infiltration. A lipid panel revealed an LDL of 18 mg/dl, total cholesterol (TC) of 91 mg/dl, triglyceride (TG) of 202 mg/dl, HDL of 33 mg/dl, apoAI of 127 mg/dl (110-205) and apoB of 47 mg/dl (55-140), while on simvastatin 40 mg daily. An earlier lipid panel obtained while patient was not on cholesterol lowering medication showed: LDL 56 mg/dl, TC 151 mg/dl, TG of 326 mg/dl, and HDL of 29 mg/dl. Prior cardiac catheterization was notable for the absence of atherosclerosis. Cardiac CT obtained during his hospitalization revealed minimal coronary artery calcification only in the LAD region. The dyslipidemia was suggestive of hypobetalipoproteinemia.[br]Conclusion: While amyloidosis of the adrenal gland has rarely been associated with adrenal insufficiency due to bilateral deposition, an association with hyperaldosteronism has not been previously reported. We hypothesize that either an adrenal nodule or hyperplasia was masked by the amyloid, or this patient may have had a genetic mutation resulting in excess aldosterone production. Whether the PA, lipid abnormality, and amyloidosis in this patient are related is unknown at this time. However, it is possible that the low LDL was cardio-protective in this diabetic patient with PA.[br][br]Nothing to Disclose: MR, RS, JB, SA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 541 193 1480 SUN-455 PO45-02 Sunday 1277 2012


1274 ENDO12L_SUN-456 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Isolated Metastatic Renal Cell Carcinoma Masquerading as an Adrenal Adenoma Jason G Daily, Robert A Vigersky Walter Reed National Military Medical Center, Bethesda, MD Background: Adrenal metastases from a variety of primary cancers are seen in up to 27% of cases, most commonly from breast and lung. Renal cell carcinoma rarely metastasizes to the adrenal glands and is usually associated with widespread disease. Isolated metastatic renal cell carcinoma to the contralateral adrenal glands occurring after initial nephrectomy is even rarer (1,2). We report a 36-year-old woman with an isolated metastatic renal cell carcinoma to the right adrenal gland.[br]Clinical Case: A 36-year-old woman was referred to the Endocrine clinic for evaluation of a 5 cm right adrenal mass. She was diagnosed with renal cell cancer in 2007 after an evaluation for hematuria that demonstrated a 10cm left kidney mass on CT. She underwent a left nephrectomy and adrenlectomy with the pathology confirming clear cell renal cell carcinoma. She had no evidence of possible metastasis until routine radiographic surveillance at another institution found a 1.5 cm right adrenal nodule in 2010. However, an evaluation was not performed at that time. In November 2011, she had an MRI of the abdomen that demonstrated that the right adrenal mass had increased in size to 5.7 x 4.9 cm. A fine-needle aspiration biopsy demonstrated malignant cells compatible with renal cell cancer. On initial evaluation, she denied a history of hypertension, palpitations, headache, pallor or sweating, but did report a history of unintentional weight loss. Physical exam was notable for normal blood pressure and pulse with no clinical evidence of hypercortisolism. Biochemical evaluation did not show any evidence of functionality of the lesion. She underwent a whole body F18-FDG PET scan which showed a heterogeneous hypermetabolic activity within the right adrenal mass and no other hypermetabolic areas to suggest metastatic disease. She then underwent a right adrenalectomy and a wedge resection of the liver without any complications. The pathology confirmed clear cell renal cancer. Gluco- and mineralocorticoid replacement was initiated post-operatively.[br]Conclusion: Isolated adrenal lesions are most often benign adenomas particularly when they are less than 2cm as was the case with our patient[apos]s presentation one year prior to our evaluation. However, this case illustrates that in the setting of a previous diagnosis of renal cell carcinoma, a nodule as small as 1.5 cm in the contralateral adrenal should be considered a metastasis and prompt an aggressive and rapid evaluation.[br][br](1) Saitoh H, Nakayama M, Nakamura K, and Satoh T. Distant metastasis of renal adenocarcinoma in nephrectomized cases. Journal of Urology. 1982;128:1092-1095. (2) Kessler O, Mukamel E, Weinstein r, Gayer E, Konichezky M, Servadio C. Metachronous renal cell carcinoma metastasis to the contralateral adrenal gland. Urology. 1998;51:539-543.[br][br]Nothing to Disclose: JGD, RAV 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 564 193 1481 SUN-456 PO45-02 Sunday 1278 2012


1275 ENDO12L_SUN-457 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Adrenal Metastases from Endometrial Carcinoma, a Rare Occurrence Syeda Sadia Zaidi, Vipul T Lakhani Vanderbilt University Hospital, Nashville, TN The spread of gynecologic neoplasms to the adrenal glands is a rare occurrence. Here we describe the unusual case of bilateral adrenal gland metastasis from endometrial carcinoma.[br]A 75 year old woman with hypothyroidism presented with postmenopausal bleeding. She reported lower abdominal pain, decrease in appetite, 80 pound unintentional weight loss and fatigue over the previous few months. Family history was significant for gastric cancer in her mother. The blood pressure was normal. Gynecologic exam showed uterus [sim] 10 weeks in size. The remainder of her exam was unremarkable. Complete blood count and chemistries were normal. She underwent dilatation and curettage which revealed endometrial adenocarcinoma. Tumor marker CA-125 was normal at 5.1 U/ml (normal [lt] 35). Computed tomography of abdomen and pelvis showed bilateral thickening of adrenal glands and 6.3 x 7.5 cm uterine mass. Patient underwent laparoscopic hysterectomy, bilateral salpingo-oophorectomy and lymph node dissection. The histopathology revealed endometrial adenocarcinoma stage 1B FIGO grade 3 (pT1B, pN0, pM0). She received adjuvant chemotherapy and radiotherapy. CT chest done later showed pulmonary emboli bilaterally and anticoagulation was started. On 6 month follow up visit, the patient reported mild right upper abdominal pain with radiation to back, weakness and nausea and vomiting. A 6.5 x 4.5 cm right adrenal mass was noted on CT, suspicious for metastatic disease. 24 hour urine metanephrines, DHEA, testosterone, cortisol stimulation test, aldosterone and plasma renin activity levels were all normal. TSH and ACTH were elevated at 5 and 129 respectively. Due to the rapid increase in size of adrenal masses and worsening of abdominal pain there was a concern of adrenal hemorrhage. Inferior vena cava filter was placed and coumadin discontinued. Her condition worsened clinically. PET scan revealed intense uptake in bilateral adrenal glands consistent with metastasis, with right adrenal mass measuring 7.9 x 5.7 cm and left 1.4 x 1.2 cm. A CT guided biopsy of right adrenal mass confirmed necrotic carcinoma favoring endometrial origin. Repeat cortisol stimulation was suboptimal and prednisone was started with some improvement in her symptoms. Chemotherapy was planned for her metastatic endometrial carcinoma, however she passed away prior to that.[br]It is important to rule out adrenal insufficiency in patients with adrenal metastasis and adrenal hemorrhage can be a life threatening complication.[br][br]Nothing to Disclose: SSZ, VTL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2097 193 1482 SUN-457 PO45-02 Sunday 1279 2012


1276 ENDO12L_SUN-458 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Case Report: Familial Glucocorticoid Deficiency Associated with Familial Focal Segmental Glomerulosclerosis Nanik Ram Khatwani, Najmul Islam Aga Khan University Hospital, Karachi, Pakistan An eight month old boy presented with increased genital pigmentation. Initial investigation revealed that he was glucocorticoid deficient and was started on hydrocortisone and fludrocortisones with a diagnosis of Addison[apos]s disease. Later fludrocortisone was withdrawn and he was diagnosed to have isolated glucocorticoid deficiency. He later developed focal segmental glomerulosclerosis for which he underwent renal transplantation at the age of five years. His two siblings and a first degree cousin also have isolated glucocorticoid deficiency, with one of the sibling died due to renal failure secondary to focal segmental glomerulosclerosis. Now at the age of twelve years, this boy is doing well on hydrocortisone treatment. Reviewing the literature, this is the first case of familial glucocorticoid deficiency associated with familial focal segmental glomerulosclerosis.[br][br]Nothing to Disclose: NRK, NI 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1221 193 1483 SUN-458 PO45-02 Sunday 1280 2012


1277 ENDO12L_SUN-459 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Extreme Sporadic Glucocorticoid Resistance Syndrome Discordant to the Glucocorticoid Receptor Genotype Aikaterini A Nella, Kerri A Kissell, Tomoshige Kino, Charalambos Lyssikatos, Bert E Bachrach, Meg Keil, Constantine A Stratakis, George P Chrousos National Institute of Child Health and Human Development, Bethesda, MD; University of Missouri Hospital, Columbia, MO; University of Athens, Athens, Greece Background: Generalized glucocorticoid resistance syndrome is a rare sporadic or familial condition defined by partial insensitivity to glucocorticoids and usually ascribed to glucocorticoid receptor (GR) gene mutations. Impaired glucocorticoid feedback results in hypersecretion of CRH, AVP, ACTH, and overproduction of adrenal cortisol, mineralocorticoid and androgenic compounds. Typically, dexamethasone doses in the order of 1-3mg/day are sufficient to suppress excessive ACTH and cortisol release, and prevent the manifestations of hyperaldosteronism, hyperandrogenism and impaired fertility.[br]Clinical case: We report a 12-year-old Caucasian girl with extreme glucocorticoid resistance in light of a heterozygotic point mutation of the GR gene. She presented at the age of 2 years 10 months with seizures, hypoglycemia, hypokalemia, hypertension and premature pubarche with mild clitoromegaly. Circulating ACTH, cortisol and adrenal androgen concentrations were markedly elevated, and bone age was advanced. Her GR had a heterozygotic missense mutation at nucleotide position 2141 (exon 8), which resulted in a defective receptor (GR[alpha]R714Q) with dominant negative activity on the wild-type GR transactivation (1). Initially, dexamethasone doses of 1-2.5mg/day suppressed urinary free cortisol (UFC) excretion, restored serum electrolytes and slowed bone age advancement.[br]Over time, hypertension control has required increasing doses of dexamethasone and 4 antihypertensives, which have not prevented end-organ damage (brain micro-infarcts and a 4.5-mm MCA aneurysm). Despite extremely high dexamethasone doses (14mg/day), the highest ever used for similar patients, circulating ACTH (209 pg/ml, nl 8-46), cortisol (40.7 [mu]g/dl, nl 5-25) androstenedione (545 ng/dl, nl 80-240), and UFC (240 [mu]g/m2/day, nl 0-20) levels remain elevated. On MRI, pituitary gland is hyperplastic with a 5-mm adenoma. Diurnal ACTH and cortisol variation is preserved and there are no stigmata of Cushing syndrome. Serum dexamethasone levels are [gt]1000ng/dl, excluding fast dexamethasone clearance.[br]Conclusions: We present a patient with extreme glucocorticoid resistance, not fully explained by her heterozygotic GR gene mutation. Additional hereditary and/or acquired defects in the regulation of tissue glucocorticoid actions could explain the severity of the syndrome. Detailed molecular karyotype and whole exome sequencing are currently performed to elucidate the genomic cause(s).[br][br](1) Nader N, Bachrach BE, Hurt DE, Gajula S, Pittman A, Lescher R, Kino T. A Novel Point Mutation in Helix 10 of the Human Glucocorticoid Receptor Causes Generalized Glucocorticoid Resistance by Disrupting the Structure of the Ligand-Binding Domain. J Clin Endocrinol Metab 2010; 95(5):2281-85.[br][br]Nothing to Disclose: AAN, KAK, TK, CL, BEB, MK, CAS, GPC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 465 193 1484 SUN-459 PO45-02 Sunday 1281 2012


1278 ENDO12L_SUN-460 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) A Newborn with 45,X/46,XY Ovotesticular Disorder of Sexual Development, a Jumping Translocation of Chromosome Y and Genetically Confirmed Congenital Adrenal Hyperplasia Sonal R Chandratre, Lei Zhang, Linda D Cooley, Bernarda E Strauss, Steven L Olsen, Laura Zapapas, Brandi Martin, Jill Jacobson Children[apos]s Mercy Hospitals and Clinics, Kansas City, MO; University of Missouri-Kansas City School of Medicine, Kansas City, MO; Children[apos]s Mercy Hospitals and Clinics, Kansas City, MO; Children[apos]s Mercy Hospitals and Clinics, Kansas City, MO; Children[apos]s Mercy Hospitals and Clinics, Kansas City, MO We describe a 39 week, 2830 g, AGA neonate who presented with genital ambiguity. Mother[apos]s history revealed polycystic ovarian syndrome, but no other androgen exposure. Family history was negative for consanguinity and infantile deaths. Height was at the 8th percentile. BP was normal. The genitourinary exam revealed a phallic structure measuring 2 cm in length with a blind dimple on the glans and a urethral opening ventrally at the base. The clitoral index was 0.8, and anogenital ratio was 0.83. No gonads were palpable. No other abnormalities were noted on the physical exam. Chromosome analysis revealed mosaicism with three cell lines: 45,X[8]/45,X,tas(Y;16) (p11.32;p13.3)[8]/45,X,tas(Y;8)(p11.32;p23.3[4]. Chromosome Y showed telomere association (tas) with chromosomes 8 and 16 in different cell lines, a [ldquo]jumping translocation[rdquo]. FISH analysis was confirmative. Meanwhile, the state newborn screen returned suspicious for CAH. Further testing revealed normal electrolytes, a significantly elevated unstimulated 17-hydroxyprogesterone (2,800 ng/dl), plasma renin activity of 29.8 ng/ml/hr, and total testosterone level of 808 ng/dl. Thyroid function tests were normal. LH and FSH were undetectable. Hydrocortisone and fludrocortisone were initiated. A care conference with the family and a multidisciplinary team was held. The family had bonded to the infant as a female. The team concurred with the female gender assignment but discussed gender dysphoria with the family and recommended against early clitoral reduction surgery. The infant underwent cystoscopy, vaginoscopy and laparoscopy on day nine of life. A normal-appearing uterus, vagina and cervix were seen. Fallopian tubes were seen, and streak gonads were removed and sent for analysis. Testosterone fell to 89 ng/dL after five days of glucocorticoids but prior to gonadectomy. The surgical pathology confirmed an ovotestis on the right and an ovary on the left. Chromosome analysis of both gonads showed the same mosaicism for two cell lines: 45,X/45,X,tas(Y;8). Genetic testing for 21-hydroxylase (OH) deficiency was performed. The patient was homozygous for 21-OH deficiency, displaying two separate mutations previously associated with 21-OH deficiency. She continues to follow up in our endocrine and multidisciplinary clinic. To the best of our knowledge, this is the 3rd reported case of a constitutional chromosome Y [apos][apos]jumping translocation[apos][apos]. This is the first reported case with co-existing CAH and ovo-testicular DSD.[br][br]Nothing to Disclose: SRC, LZ, LDC, BES, SLO, LZ, BM, JJ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 665 193 1485 SUN-460 PO45-02 Sunday 1282 2012


1279 ENDO12L_SUN-461 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Triple A Syndrome: Short-Term Response to the Antioxidant N-Acetylcysteine Treatment in a Child with a Novel [italic]AAAS[/italic] Gene Mutation and Increased Oxidative Stress Maria Candida Barisson Villares Fragoso, Rodrigo Bomeny de Paulo, Paula Awake Rosa, Maria Heloisa Massola Shimizu, Antonio Carlos Seguro, Uenis Tannuri, Katrin Koehler, Angela Hubner, Ana Claudia Latronico, Ivo Jorge Prado Arnhold, Berenice Bilharinho Mendonca Hospital das Clinicas da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Technical University of Dresden, Dresden, Germany Background: Triple A syndrome is an autosomal recessive disorder characterized by alacrima, achalasia, adrenal insufficiency, autonomic dysfunction, and neurodegeneration. Mutations in the AAAS gene encoding the protein ALADIN have been reported in these patients. Patient[apos]s fibroblasts exhibit increased oxidative stress that might be related to progressive neurodegeneration. Previous studies have shown that N-acetylcysteine (NAC) protects renal function in rats and humans with kidney injuries associated with increased oxidative stress. Patient: We studied a 9 yr-old boy born to consanguineous parents. At 1.5 years he had neurodevelopmental delay, repeated otitis and gastroesophageal reflux. At 3 years, investigation of episodes of hypoglycemia and seizures led to diagnosis of primary adrenal insufficiency and treatment with glucocorticoids. At age 8 years, he had abdominal pain and jejuno-ileal intusseption was visualized at laparotomy. After surgery, nausea and vomiting after eating persisted. A swallowing videogram showed distal esophageal stenosis and endoscopy was suggestive of achalasia. The lower esophageal sphincter was dilated with baloon twice. At age 9 years, height was 109 cm (SDS-3.7), weight 18.5 Kg (SDS -1.62) and his skin was extremely itchy and dry and his hair pale and dry. Schirmer test showed alacrima. Manometry confirmed achalasia and the patient was submitted to videolaparoscopic Heller myotomy. Methods: We sequenced the AAAS gene and measured the serum levels of thiobarbituric acid reactive substances (TBARS), markers of lipid peroxidation. Results: A new homozygous missense mutation in exon 6 c.523G[gt]T, p.Val175Phe was indentified in the patient; his mother was heterozygous for the same mutation. Basal TBARS was increased (8.72 nM/ml) in comparison to normal children (1.61[plusmn]0.04 nM/ml, n=43). NAC was introduced (400 mg twice/day) and TBARS levels reduced to 1.60 nM/ml after 3 months and were kept into normal levels after 5 months (1.79 nM/ml). After 5 months of surgery and NAC treatment, decreased vomiting and catch-up growth (growth velocity = 8.8 cm/yr) were observed as well as an improvement of skin and hair condition. Conclusion: We describe a boy with Triple A syndrome due to a novel mutation in AAAS gene with elevated oxidative stress which diminished after treatment with N-acetylcysteine. Long-term follow up is needed to confirm improved outcome to allow the recommendation of NAC treatment to patients with Triple A syndrome.[br][br]Journal of Surgical Research 171, e179[ndash]e180 (2011) doi:10.1016/j.jss.2011.07.050Journal of Surgical Research 171, e179[ndash]e180 (2011) doi:10.1016/j.jss.2011.07.050.[br][br]Nothing to Disclose: MCBVF, RBdP, PAR, MHMS, ACS, UT, KK, AH, ACL, IJPA, BBM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 820 193 1486 SUN-461 PO45-02 Sunday 1283 2012


1280 ENDO12L_SUN-462 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Intra-Articular Glucocorticoid Injections as a Cause for Adrenal Insufficiency Brooke Sanders, Reza Nazemi, John David Carmichael Cedars-Sinai Medical Center, Los Angeles, CA; Cedars-Sinai Medical Center, Los Angeles, CA Background: Intra-articular steroid injections (IASI) can be associated with decreased release of endogenous cortisol, but are rarely thought to cause adrenal insufficiency (AI). We present five patients who presented to our center with AI, secondary to IASI.[br]Clinical Cases: Five patients (3F/2M, age range: 39-70 yrs) presented with fatigue, lethargy, shortness of breath, truncal obesity, headaches. All five had a recent history of IASI for various indications. Initial Cortrosyn stimulation tests revealed insufficient responses: (mean baseline cortisol 0.6 [plusmn] 0.738 mcg/dL; mean peak cortisol 6.76 [plusmn] 3.74 mcg/dL.) Patients each received average 3.8 injections (range: 2-6) of triamcinolone acetonide to the knee, ankle, lumbar spine (2 patients), and cervical spine. All five patients were diagnosed with AI secondary to suppression of the hypothalamic-pituitary-adrenal (HPA) axis and treated with hydrocortisone replacement therapy. Follow-up Cortrosyn stimulation tests were performed periodically (6-12 weeks) to assess for return of HPA axis function (peak cortisol [gt]18 mcg/dL.) The median ([plusmn]SD) time to recovery was 16 [plusmn] 38 wks (range: 6-88 wks). None of the patients in this series reported receiving IASI until directly asked. None knew the contents of the injections. All were seen and evaluated for presenting complaints by 2 or more physicians prior to diagnosis.[br]Discussion: While exogenous glucocorticoid administration is a well known cause of HPA axis suppression and AI, IASI are not recognized as commonly causing AI. Many barriers exist to diagnosing patients with AI secondary to AISI. As illustrated in our series, patients often do not recall receiving IASI and do not consider them part of their current medical regimen. Patients rarely develop cushingoid features, further obscuring the etiology of AI. Severity of cushingoid features, extent of suppression, and time to recovery of the HPA axis varies among individuals and may be due to multiple factors. There is great variability in physiologic response to steroid injections due to glucocorticoid receptor (GR) polymorphisms, epigenetics, and interactions with other prescription drugs, specifically PARP (poly ADP ribose polymerase) inhibitors. Well-known factors such as dose, duration of treatment, and drug half-life also contribute to varying responses.[br]Conclusion: This series illustrates an under-recognized etiology of isolated AI and highlights barriers to diagnosing subjects with AI secondary to IASI.[br][br]Nothing to Disclose: BS, RN, JDC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2194 193 1487 SUN-462 PO45-02 Sunday 1284 2012


1281 ENDO12L_SUN-463 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) A Novel Indel Mutation in the [italic]AAAS[/italic] Gene Causes Triple A Syndrome with Delayed Puberty Katrin Koehler, Haidar A Bustanji, Bashar N Sahar, Angela Huebner, Dana Landgraf, Hanan A Hamamy, Kamel M Ajlouni Technical University Dresden, Dresden, Germany; National Center for Diabetes Endocrinology and Genetics, Amman, Jordan; Geneva University Hospital, Geneva, Switzerland Background: The triple A syndrome is a rare autosomal recessive disease combining severe adrenal insufficiency with neurological impairment. Hallmark of the disease is the peculiar combination of adrenocorticotropic hormone (ACTH) resistant adrenal failure, achalasia and alacrima. Usually patients with this syndrome enter puberty at an appropriate age. The triple A syndrome is caused by mutations in the [italic]AAAS [/italic]gene on chromosome 12q13, which encodes the 60 kDa WD repeat-containing nucleoporin ALADIN.[br]Clinical case: Here we report on a 17 year-old boy presenting with delayed puberty, a four years history of fatigue and muscle weakness. He had achalasia, alacrima, skin and mucosal hyperpigmentation. Hormonal assessment revealed isolated glucocorticoid deficiency. Clinical diagnosis of triple A syndrome was confirmed by sequencing the [italic]AAAS[/italic] gene.[br]Molecular studies: Analysis of the genomic DNA of the patient revealed a homozygous 1648 bp deletion including a part of intron 7, exons 8-10 and intron 10. Instead of this sequence an insertion of an unknown 12 bp nucleotide sequence TGAGGCCTGCTG is located ahead of exon 11. The splice acceptor site of intron 10 was also deleted. With RT-PCR analysis on RNA from patient and control cells we can show that the deletion leads to a splicing defect. According to the nomenclature of den Dunnen et al. this novel mutation is referred to as g.16166_17813delinsTGAGGCCTGCTG (NG_016775). As in the heterozygous parents the deletion of exons 8-10 on genomic DNA level was not detected by sequencing the entire coding region including exon-intron boundaries of the [italic]AAAS [/italic]gene, this mutation may also be overlooked in compound heterozygous patients. The pubertal delay is apparently a rare feature of triple A syndrome as most patients enter puberty at normal time. [italic]AAAS[/italic] is highly expressed in the pituitary gland which may serve as a possible explanation for the impairment of pituitary function.[br]Conclusion: This is the first report of triple A syndrome in Jordan with a novel indel mutation in the [italic]AAAS[/italic] gene and presenting with delayed puberty.[br][br]Nothing to Disclose: KK, HAB, BNS, AH, DL, HAH, KMA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1217 193 1488 SUN-463 PO45-02 Sunday 1285 2012


1282 ENDO12L_SUN-464 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) De Novo Mutation of CYP21A2 Gene in Patient with Non-Classical Congenital Adrenal Hyperplasia Jessica Schratter, Manjula Chatterjee St George[apos]s University, True Blue, Grenada; The Brooklyn Hospital Center, Brooklyn, NY Background: Late-onset (or non classical) congenital adrenal hyperplasia (NCAH) is an autosomal recessive condition resulting most commonly from partial deficiency of CYP21A2 gene (encoding 21-hydroxylase enzyme in adrenal steroidogenesis). The degree of enzymatic inactivity determines the severity of the illness. As the availability of molecular testing increases, more mutations are being described in the literature. This report depicts a young girl who was diagnosed with NCAH and was found to have compound heterozygous mutation for the p.V281L and a novel p.Glu352Argfs*30 mutation in the CP21A2 gene resulting in no functional activity of the gene.[br]Methodology: Assays determined by Quest Diagnostics Nichols Institute, San Juan Capistrano, were used to detect the common mutations. This was followed by PCR amplification and complete sequence analysis of the amplified DNA.[br]Clinical case: Index patient is a previously healthy 13-year-old Hispanic female who presented with secondary amenorrhea. She had no other signs of hyperandrogenization like hirsutism, acne, male pattern baldness, or clitoromegaly. There was no family history of infertility or consanguinity. Work up revealed elevated basal and ACTH-stimulated 17-hydroxyprogesterone (17-OHP) and subnormal stimulated serum cortisol level. Molecular analysis of CYP21A2 gene revealed one copy each of p.V281L mutation, a common form associated with NCAH in which a valine is substituted to leucine at amino acid position 282 due to G[gt]T at nucleotide g.1688 in exon 7, and a novel p.Glu352Argfs*30 mutation. Latter results from a single nucleotide duplication of C at g.2097 in exon 8. This shifts the reading frame of CYP21A2 mRNA resulting in premature protein truncation. As the two mutations were found on two different copies of CYP21A2 gene, there was no functional activity of CYP21A2 gene detected. Therefore, based on the nature of the change, the p.Glu352Argfs*30 mutation can be expected to be associated with a more severe form of NCAH.[br]Conclusion: We describe a novel mutation, p.Glu352Argfs*30, in CYP21A2 gene in a teenager which has not been described previously in literature. This adds to the existing pool of genetic polymorphisms seen in NCAH, and perhaps simple-virilizing congenital adrenal hyperplasia, thereby, providing us with better understanding of genotype/phenotype correlation leading to appropriate genetic counseling.[br][br]Nothing to Disclose: JS, MC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 407 193 1489 SUN-464 PO45-02 Sunday 1286 2012


1283 ENDO12L_SUN-465 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Large Ovarian Cysts in Patients with Disorders of Steroidogenesis [mdash] Clinical Course and Optimal Treatment Akiko Yamamoto, Satsuki Nishigaki, Yusuke Mizuno, Kengo Miyashita, Masahiro Noda, Yasuhiro Naiki, Reiko Horikawa National Center for Child Health and Development, Okura,Setagaya-ku, Japan Background: Ovarian cyst is common findings in females affected by steroidogenic disorders.[br]Patients: We experienced 4 patients having large ovarian cysts.[br]Case1: 10 years old girl with PORD. At 9 years old, she had abdominal pain and severe constipation. MRI revealed a large ovarian cyst (60*62*75mm). Her E2, LH, FSH level were 22pg/ml(0-31), 27.34mU/ml([lt]0.05-1.31), 10.73mU/ml([lt]0.12-5.35), respectively. She was treated with GnRH analog without complications. Abdominal pain almost diminished. After 1month of treatment, her E2, LH, FSH level significantly decreased and her cyst became smaller in size.[br]Case 2: 38 years old woman with CLAH. From 22 years of age, she complained abdominal pain and has been followed by imaging studies. After 3 years of f/u, pain worsened and ultrasonography and MRI showed ovarian cyst of 62*57*67mm cystic lesion, with no solid component. After 1 year treatment with GnRH analog, cyst volume was decreased to 36*17*40mm, and the abdominal pain has gone.[br]Case 3: 22 years old woman with PORD. At the age of 13, she had sudden dysurination. MRI showed large ovarian cyst (74.2*55.3mm). From images, the ovarian tumor was suspected of malignancy, and the tumor was resected. The pathology was serous cystadenoma.[br]Case 4: 39 years old women with CLAH. She has received regular follow-up by gynecologist for polymenorrhea and intermittently treated with gestagen. Small unilateral ovarian cyst was found and followed for 3 years. She did not show any symptoms until when she had acute intolerable abdominal pain. The ovarian cyst has enlarged to 50*40mm. Torsion was suspected, however, the pain was transient and the ovarian cyst was not operated. The ovarian lesion is under f/u by gynecologist.[br]Discussion: Ovarian cyst is common complication in patients with steroidogenic disorders, whereas extremely large cyst tend to occur in CLAH, PORD, and aromatase deficiency. It can cause not only abdominal pain but also severe constipation and dysurination. With its stalk torsion, it may cause abdominal emergency that needs surgery. In present cases with PORD and CLAH, ovarian cysts have enlarged after puberty. In 2 cases, we tried GnRH analog, which was effective for remission of large cysts and improved symptoms without complications. The mechanism of cyst formation in PORD and CLAH may not be the same, however, our observation suggested the elevation of gonadotropin levels play a key role for the formation of large ovarian cyst in these two disorders.[br][br]Nothing to Disclose: AY, SN, YM, KM, MN, YN, RH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1375 193 1490 SUN-465 PO45-02 Sunday 1287 2012


1284 ENDO12L_SUN-466 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Edema, Enigma: Secondary to 11 B-Hydroxysteroid Dehydrogenase Type 2 Inhibition by Sweetener [apos]Stevia[apos] Salina Esmail, Udaya M Kabadi Private Practice, Upton, CA; University of Iowa, Iowa City, IA; Veterans Affairs Medical Center, Des Moines, IA Edema, Hypertension and Hypokalemia occur with inhibition of 11 B-Hydroxysteroid Dehydrogenase Type 2 (11B-HSD2) by chronic Licorice ingestion. Herein is 1[sup]st[/sup]report of Edema and Prehypertension induced by 11B-HSD2 inhibition by sweetener Stevia.[br]Case Report:32 year old Caucasian woman was seen for edema of over 6 months. Cardiac,Renal and Hepatic causes were excluded by normal ECG and Echocardiogram; normal serun urea nitrogen (SUN),creatinine (Creat), without proteinuria and normal liver enzymes.Furosemide and KCL were discontinued 3weeks before visit. At the clinic visit, edema in hands,feet and face was confirmed. BP was 138/88 mm Hg with regular pulse,74/min. Heart, lung, abdomen and neurological Examinations were unremarkable.CBC,Urinalysis, SUN,Creat,Proteins,liver enzymes,Calcium,Phosphorus and lipids were normal. prior to and on withdrwal of Stevia. Serum Na,141mM/L; K, 3.5mM/L; Cl,95mM/L and HCO[sub]3[apos][/sub]30mM/L and 24 Hour Urine Na 120mM and K, 102mM.Serum Aldosterone,[lt] 1.6 ng/dl (low), Renin Activity,0.4ng/ml/hr(low). Hence, AM serum Cortisol and Cortisone and 24 hour urine Na, K, Free Cortisol and Free Cortisone were determined.AM Cortisol 8.2 ug/dl (Normal); Cortisone,0.2 ug/dl (low);24 hour urine Na,120mM; K,102mM; Free Cortisol,109 mcg (high) and free cortisone,1.4ug (low).suggested 11B-HSD2 inhibition. Hence, subject was questioned regarding Licorice ingestion or chewing tobacco containing Licorice. She denied. On further inquiry,she reported use of sweetener Stevia for 9 months prior to visit. She was asked to abstain from Stevia and return to clinic at 3 months.At the visit,she reported remission of edema 2 weeks after discontinuation of Stevia.BP was 118/74mmHg with a regular pulse,76/min. Rest of examination was unremarkable.Marked changes were serum K,4.3 mM/l; Cl,100 mM/L;HCO[sub]3[/sub],25 mM/l;Aldosterone,6.8ng/dl;Renin Activity,2.2ng/ml/hr; cortisol,10.7ug/dl; cortisone,2.3 ug/dl;24 hour urineNa,160 mM/l;K,64mM/l;Free Cortisol,44ug;Free Cortisone,8.0 ug.[br]Conclsion: Onset of edema, prehypertension,mild lowering of Serum K, decline in aldosterone and renin activity, an increase in Cortisol/Cortisone ratio on chronic ingestion of sweetener Stevia with normalization of laboratory abnormalities and total remission of clinical manifestations following discontinuation of Stevia indicate a role of Stevia in induction of edema and Prehypertension via reduced conversion of Cortisol into Cortisone by inhibition of 11 B-Hydroxysteroid Dehydrogenase Type 2.[br][br]Nothing to Disclose: SE, UMK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 253 193 1491 SUN-466 PO45-02 Sunday 1288 2012


1285 ENDO12L_SUN-467 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Pseudohypoaldosteronism Presenting with Thrombocytosis and Bilateral Pneumothoraces in an Infant: Case Report and Literature Review Asma Javed, Jennifer M Leonard, Cramer H Carl, Kumar Seema, Kirmani Salman, Chad K Brands Mayo Clinic, Rochester, MN; All Children[apos]s Hospital Johns Hopkins Medicine, St Petersburg, FL [bold]Background:[/bold] PHA-1 is a salt wasting syndrome caused by peripheral resistance to aldosterone. The autosomal recessive form is life-threatening and due to mutations in the epithelial sodium channel. PHA-1 autosomal dominant form is caused by a mutation in the mineralocorticoid receptor and manifests in infancy with failure to thrive. This gradually resolves reflecting the process where infants become less reliant on aldosterone.[br][bold]Clinical Case[/bold]: A 5 week old Caucasian male infant presented with vomiting and failure to thrive. Initial serum chemistries showed mild hyponatremia (135 mMol/L N 135-145mMol/L), hyperkalemia (6.2 mMol/L N 3.5-5.1mMol/L), hypercalcemia (11.8mg/dl N9.6-10.8 mg/dl) and thrombocytosis (937 X 10 (9) N 150-450 X10 (9)). His weight was 2.72 kg, below birth weight of 2.8 kg. The child was born term and diagnosed with bilateral pneumothoraces, patent ductus arteriosus and atrial septal defect soon after birth. He developed emesis and failure to gain weight. The patient[apos]s maternal uncle had failure to thrive which resolved with salt supplementation. The patient[apos]s investigations included normal urinalysis, renal ultrasound, upper GI and voiding cystourethrogram. He had normal AM Cortisol (3.8 mcg/dl N 2.8-23mcg/dl), 17 hydroxyprogesterone (80 ng/dl N [lt] 110 mcg/dl) and DHEA ([lt] 1.0 ng/ml N [lt]2.9 ng/ml) and markedly elevated serum aldosterone (680 ng/dl N 6.5-86 ng/dl) and renin activity (330 N 4.6 ng/ml/hr). These results combined with a normal epithelial sweat chloride test suggested a diagnosis of PHA-1, autosomal dominant form. He was started on every other day 0.6mg/kg/day Sodium Polystyrene Sulfonate (SPS) and daily 4meq/day sodium chloride supplementation with improvement. DNA testing for mutations in the mineral corticoid receptor gene (NR3C2, also known as nuclear receptor subfamily C2 gene) revealed a novel mutation denoted c.2919_2920insG (p.975Efs1004x). The patient was followed every six weeks with repeat serum chemistries. SPS for hyperkalemia was stopped at 7 months of age and sodium supplementation stopped at 11 months of age. At one year, he was thriving. The hypercalcemia and thrombocytosis resolved without intervention.[br][bold]Conclusion [/bold]Hyponatremia and hyperkalemia in infancy can represent a variety of renal and genetic disorders. We describe a case of PHA-1, autosomal dominant form, presenting with failure to thrive, salt wasting, hyperkalemia and thrombocytosis at five weeks of life, preceded by bilateral pneumothoraces at birth.[br][br]1. Geller, D.S., J. Rodriguez-Soriano, A. Vallo Boado, S. Schifter, M. Bayer, S.S. Chang, and R.P. Lifton. 1998. Mutations in the mineralocorticoid receptor gene cause autosomal dominant. pseudohypoaldosteronism type I. Nat Genet. 19:279-81. 2. Chang, S.S., S. Grunder, A. Hanukoglu, A. Rosler, P.M.Mathew,. I. Hanukoglu, L.Schild, Y. Lu, R.A. Shimkets, C. Nelson-Williams,. B.C. Rossier, and R.P. Lifton1996. Mutations in subunits of the. epithelial sodium channel cause salt wasting with hyperkalaemic. acidosis, pseudohypoaldosteronism type 1. Nat Genet. 12:248-53. 3. Riepe, F.G. 2009. Clinical and molecular features of type 1. pseudohypoaldosteronism.Horm Res. 72:1-9. 4. Bonny, O., and B.C. Rossier. 2002. Disturbances of Na/K. balance: pseudohypoaldosteronism revisited. J Am Soc Nephrol. 13:2399-414. 5. Chan KW, Kaikov Y, Wadsworth LD1989 Thrombocytosis in. childhood: a survey of 94 patients. Pediatrics. 1989. Dec;84(6):1064-7. 6. Watanabe, T., and A. Yamazaki. 2003. Pneumothorax and. transient pseudohypoaldosteronism in an infant with. hydronephrosis. Pediatr Nephrol. 18:62-4. 7. Kerem, E., T. Bistritzer, A. Hanukoglu, T. Hofmann, Z. Zhou,. W. Bennett, E. MacLaughlin, P. Barker, M. Nash, L. Quittell, R. Boucher, and M.R. Knowles. 1999. Pulmonary epithelial sodium. channel dysfunction and excess airway liquid in. pseudohypoaldosteronism. N Engl J Med. 341:156-62.[br][br]Nothing to Disclose: AJ, JML, CHC, KS, KS, CKB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 847 193 1492 SUN-467 PO45-02 Sunday 1289 2012


1286 ENDO12L_SUN-468 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Hypergonadotropic Hypogonadism in a Female with Non-Classical Congenital Adrenal Hyperplasia Katherine Velez, Rona Sonabend Texas Children[apos]s Hospital, Baylor College of Medicine, Houston, TX [bold]Background [/bold][br]Menstrual irregularities are common in female patients with classical and non classical Congenital Adrenal Hyperplasia (CAH). Hyperandrogenemia in these patients suppresses Luteinizing Hormone (LH) surge leading to anovulation. Although this is the presumed mechanism for menstrual irregularities in females with CAH, additional etiologies should be considered. We report a case of an adolescent female with amenorrhea secondary to non classical CAH and primary ovarian failure.[br][bold]Clinical Case[/bold][br]We describe a 15 years old Hispanic female with primary amenorrhea and delayed puberty. Physical exam revealed Tanner 1 breasts, Tanner 3 pubic hair, 2 cm clitoromegaly without hirsutism or acne. Evaluation revealed a 46 XX karyotype, elevated gonadotropins (FSH 53.7 MIU/mL, LH 26.7 MIU/mL), undetectable estradiol ([lt]2 pg/mL) and inhibin B ([lt]10 pg/mL). Pelvic ultrasound revealed normal female genitalia with prepubertal structures. Further workup for clitoromegaly revealed 17 hydroxyprogesterone of 6435 ng/dL, 17 hydroxypregnenolone of 1565 ng/dL, and partial adrenal insufficiency (cortisol level 15 mcg/dL) after 250 mcg ACTH stimulation. Genetic analysis found two copies of V281L mutation significant for non classical 21 hydroxylase deficiency CAH and one copy of R356W mutation associated with classical 21 hydroxylase deficiency CAH. Treatment for CAH with maintenance hydrocortisone was initiated with resolution of elevated sex steroid precursors. However, gonadotropins remained elevated with no progression of puberty suggesting a separate diagnosis of primary ovarian failure. Estrogen replacement therapy resulted in breast development and menstrual cycles. Work up for the etiology of the ovarian failure is negative thus far.[br][bold]Conclusion[/bold][br]Primary ovarian failure as supported by elevated gonadotropins and non detectable estradiol levels appears to be the etiology of this patient[apos]s amenorrhea. This case is the first to describe a patient with non classical CAH and primary ovarian failure.[br]The possibility of a separate etiology for menstrual irregularities in patients with CAH should be investigated if despite adequate treatment symptoms persist. Future research can be directed to elucidate a genetic association between CAH and primary ovarian failure.[br][br]Nothing to Disclose: KV, RS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1955 193 1493 SUN-468 PO45-02 Sunday 1290 2012


1287 ENDO12L_SUN-469 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Adrenal Insufficiency in Patients with Hyperinsulinemic Hypoglycemia Shelly Mathur, Jasmine Boparai, Glenn Cunningham, Jose Garcia, Sanjay Mediwala, Marco Marcelli Baylor College of Medicine, Houston, TX; Michael E DeBakey Veterans Affairs Medical Center, Houston, TX; University of Texas Medical Branch at Galveston, Galveston, TX; St Luke[apos]s Episcopal Hospital, Houston, TX Background: The normal physiologic response to hypoglycemia is characterized by a rise in glucagon, catecholamines, growth hormone and cortisol. Described here are five patients with hypoglycemia due to hyperinsulinism with an inadequate cortisol response to hypoglycemia defined as cortisol [lt] 18 ug/dL (n [gt] 18 ug/dL) when glucose [lt] 45mg/dL (n 65-110mg/dL). Four out of the five patients examined achieved resolution of their adrenal insufficiency after they received surgical or medical treatment for their hyperinsulin state. Though there have been a few sporadic cases reported in the literature, this entity may be more common than previously thought.[br]Clinical Case: The age of the patients ranged from 36-66 years with 3 female and 2 male patients. Etiology of hypoglycemia included a history of Roux-en-Y gastric bypass surgery in four out of five of the patients, while one patient was diagnosed with an insulinoma. The cortisol response ranged from 4-15.5 ug/dL (n [gt] 18 ug/dL) in response to spontaneous hypoglycemia when glucose [lt] 45mg/dL (n 65-110mg/dL) based on available data in four of the patients. One of these patients was immediately started on glucocorticoid therapy based on these results. Three patients were confirmed to have adrenal insufficiency on formal testing based on cortisol [lt] 18 ug/dL (n [gt] 18 ug/dL) in response to cosyntropin and were started on glucocorticoid therapy. Only one patient tested negative for adrenal insufficiency despite the above defined inadequate cortisol response. Patients on glucocorticoids were able to discontinue their therapy (range 4.5-20 months) after they received treatment of their hyperinsulin state based on normal cortisol response (cortisol [gt] 18 ug/dL) during formal testing with cosyntropin. Three out of four of these patients received surgical treatment, while the other patient received medical treatment for their correction of their hyperinsulin state.[br]Conclusion: Hyperinsulinism and coexisting adrenal insufficiency may be more common than previously thought. Screening for adrenal insufficiency might be warranted in the setting of hyperinsulinism to determine the need for glucocorticoid therapy.[br][br]Disclosures: JG: Investigator, Aeterna Zentaris Inc. Nothing to Disclose: SM, JB, GC, SM, MM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 271 193 1494 SUN-469 PO45-02 Sunday 1291 2012


1288 ENDO12L_SUN-470 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Vitamin D Deficiency/Insufficiency in Patients with Non-Classic Adrenal Hyperplasia and Response of the Latter to Vitamin D Replacement Magalys Vitiello, Gul Bahtiyar, Alan Sacerdote Woodhull Medical [amp] Mental Health Center, Brooklyn, NY; State University of New York (SUNY) Downstate Medical Center, Brooklyn, NY; St George[apos]s University, School of Medicine, Grenada, WI; New York University School of Medicine, New York, NY [bold]Background[/bold][br]Vitamin D Deficiency/Insufficiency (VDDI) has become common worldwide in association with lifestyle changes and increased surveillance. Although its role in regulating calcium and phosphorus homeostasis is well recognized, recent studies have identified a relationship between low vitamin D levels and multiple medical conditions. VDDI is also reported in association with Type II DM, insulin resistance (IR), and polycystic ovarian syndrome (PCOS). Non-classic adrenal hyperplasia (NCAH) shares many features with PCOS including: IR, hyperandrogenism, menstrual irregularity, polycystic ovaries, and hypofertility. Many of the same insulin-sensitizing interventions that ameliorate PCOS also ameliorate the biochemical and clinical expression of NCAH including: metformin, thiazolidinediones, weight loss, and Roux-en-Y gastric bypass. Vitamin D has been shown to improve the ovulation rate of patients with PCOS. Therefore, it would not be surprising if VDDI was prevalent in NCAH patients and if Vitamin D supplementation ameliorated the expression of NCAH.[br][bold]Methods and Results[/bold][br]We reviewed the charts of 9 NCAH patients; 6 women and 3 men, aged 28 to 71 years old, all with concomitant VDDI. 25-OH-Vitamin D was measured by liquid chromatography tandem mass spectrometry (LCMSMS). Serum 25-OH-Vitamin D levels [lt] 20 ng/dl were considered deficient and levels between 20 and 30 ng/dl were considered insufficient. Vitamin D supplementation was initiated in all nine patients.[br]One patient, a 71 year old woman with a baseline serum 11-deoxycortisol measured by LCMSMS of 63 ng/dl (normal: [lt]37) and a baseline 25-OH-Vitamin D level of 29 ng/dl was treated with ergocalciferol 50,000 IU every 2 weeks. Treatment was accompanied by a fall in serum 11-deoxycortisol to [lt] 20 ng/dl.[br][bold]Conclusions[/bold][br]From our observation in this series of patients, we believe that VDDI may be more prevalent in NCAH patients than in the general adult population and based on the many clinical similarities between this condition and PCOS, Vitamin D may be an effective component of NCAH treatment. The decrease in 11-deoxycortisol levels observed in one of our patients following supplementation with vitamin D potentially offers an anecdotal occurrence of improvement of the primary condition (NCAH) which merits further analysis. Accumulated evidence from larger randomized clinical trials will be needed to confirm the therapeutic value of vitamin D supplementation for the general population of patients with NCAH.[br][br](1)Grundmann M, von Versen-H[ouml]ynck F. Vitamin D - roles in women[apos]s reproductive health? Reprod Biol Endocrinol. 2011 Nov 2;9:146. (2) Ngo DT, Chan WP, Rajendran S, Heresztyn T, Amarasekera A, Sverdlov AL, O[apos]Loughlin PD, Morris HA, Chirkov YY, Norman RJ, Horowitz JD. Determinants of insulin responsiveness in young women: Impact of polycystic ovarian syndrome, nitric oxide, and vitamin D. Nitric Oxide. 2011 Oct 30;25(3):326-30. (3)Wehr E, Pieber TR, Obermayer-Pietsch B. Effect of vitamin D3 treatment on glucose metabolism and menstrual frequency in polycystic ovary syndrome women: A pilot study.J Endocrinol Invest. 2011 Nov;34(10):757-63. (4) Li HW, Brereton RE, Anderson RA, Wallace AM, Ho CK. Vitamin D deficiency is common and associated with metabolic risk factors in patients with polycystic ovary syndrome.Metabolism. 2011 Oct;60(10):1475-81. (5) Wehr E, Trummer O, Giuliani A, Gruber HJ, Pieber TR, Obermayer-Pietsch B. Vitamin D-associated polymorphisms are related to insulin resistance and vitamin D deficiency in polycystic ovary syndrome.Eur J Endocrinol. 2011 May;164(5):741-9. (6) Strohmayer E, Via MA, Yanagisawa R. Metabolic management following bariatric surgery.Mt Sinai J Med. 2010 Sep-Oct;77(5):431-45. (7) Wehr E, Pilz S, Schweighofer N, Giuliani A, Kopera D, Pieber TR, Obermayer-Pietsch B. Association of hypovitaminosis D with metabolic disturbances in polycystic ovary syndrome. Eur J Endocrinol. 2009 Oct;161(4):575-82.[br][br]Sources of Research Support: None.[br][br]Nothing to Disclose: MV, GB, AS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 353 193 1495 SUN-470 PO45-02 Sunday 1292 2012


1289 ENDO12L_SUN-471 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) A Case of Addison Disease with Initially Normal Cortisol: When Labs Lag behind Symptoms Laura Elizabeth Hinz, Gregory Kline University of Calgary, Calgary, Canada Background[br]A number of cases of primary adrenal insufficiency have been reported in which cortisol levels are normal in the initial stages of the disease.(1-4) The introduction of assays to detect adrenal antibodies have allowed the autoimmune variant of the disease to be picked up earlier, leading to the suggestion that a drop in cortisol may be a late diagnostic marker.[br]Clinical Case[br]A 41 year old woman with known autoimmune hypothyroidism, premature ovarian failure and colitis presented to her Endocrinologist with nausea, salt craving, increased skin pigmentation and postural hypotension. Initial bloodwork revealed a normal morning cortisol of 532nmol/L (n 200-690) but an ACTH level ten times normal (113pmol/L; n [lt]11.5). Her aldosterone was 114pmol/L (n 340-1730) and renin was 6.40ng/L/s (n 0.28-1.67). Six weeks after her initial presentation, she was found to have anti-adrenal antibodies but a random cortisol of 423nmol/L, well within normal range. It was not until ten weeks after her initial symptomatic presentation that her random am cortisol was found to be [lt]1.3nmol/L yet without ever having had hyperkalemia. She was started on Hydrocortisone 10mg BID and Fludrocortisone 0.05mg OD. Four months after her first visit, her symptoms had resolved, she had gained 2.5kg and was normotensive.[br]Clinical Lessons[br]Despite classic signs and symptoms of adrenal failure and an autoimmune history, the patient retained a high-normal serum cortisol on initial investigation. The initial cortisol was sufficiently high such that further testing would not usually be considered. In the setting of adrenal antibodies, Bettrele has suggested that laboratory findings progress through four stages: 1)increased plasma renin, 2)decreased response to stimulation with cosyntropin, 3)increased ACTH and 4)decreased cortisol.(5) The timing of these changes has not been established, but our patient experienced ten weeks of symptomatic adrenal insufficiency and an elevated ACTH before the cortisol fell to a level diagnostic of adrenal insufficiency. This case illustrates the lack of sensitivity of a morning cortisol to rule out Addison[apos]s disease. Our findings corroborate Bettrele[apos]s suggestion that mineralocorticoid function can be lost before glucocorticoid function, yet without provoking hyperkalemia. We propose that an ACTH level be included in the initial work-up of suspected adrenal insufficiency, particularly in the setting of polyglandular autoimmune failure or adrenal autoantibodies.[br][br]1. Goodwin TJ, Kind P, Bogomoletz WV. Miliary tuberculosis, Addison[apos]s disease and a normal plasma cortisol level. Tubercle. 1972; 53(251-4). 2. Kong MF, Jeffcoate W. Eighty-six cases of Addison[apos]s disease. Clinical Endocrinology. 1994; 41: 157-61. 3. Soule S. Addison[apos]s disease in Africa- a teaching hospital experience. Clinical Endocrinology. 1999; 50: 115-20. 4. Dorin RI, Qualls CR, Crapo LM. Diagnosis of adrenal insufficiency. Annals of Internal Medicine. 2003; 139: 194-204. 5. Betterle C, et al. The natural history of adrenal function in autoimmune patients with adrenal autoantibodies. Journal of Endocrinology. 1988; 117: 467-475. 6. Gatti R et al. Cortisol assays and diagnostic laboratory procedures in human biological fluids. Clinical Biochemistry. 2009; 42: 1205-17. 7. Kao PC, Machacek A, Magera MJ, Lacey JM, Rinaldo P. Diagnosis of adrenal cortical dysfunction by liquid chromatography-tandem mass spectrometry. Annals of Clinical and Laboratory Science. 2001; 31(2): 199-204. 8. Wissner Greene L, et al. Adrenal insufficiency as a complication of the acquired immunodeficiency syndrome. Annals of Internal Medicine. 1984; 101(4): 497-8. 9. Falorni A et al. High diagnostic accuracy for idiopathic Addison[apos]s disease with a sensitive radiobinding assay for autoantibodies against recombinant human 21-hydroxylase. Journal of Clinical Endocrinology and Metabolism. 1995; 80: 2752-. 10. Butcher GP, Zambon M, Moss S, and Walters JRF. Addisonian crisis presenting with a normal short tetracosactrin stimulation test. Postgraduate Medical Journal. 1992; 68: 465-6. 11. Patel S, Selby C, Dornan TL, Jeffcoate WJ. Response to Addisonian crisis presenting with a normal short tetracosactrin stimulation test. Postgraduate Medical Journal. 1993; 69: 808. 12. Gonzalez-Gonzalez JG, et al. A high-sensitivity test in the assessment of adrenocortical insufficiency: 10ug vs. 250ug cosyntropin dose assessment of adrenocortical insufficiency. Journal of Endocrinology. 1998; 159: 275-280. 13. Hurel SJ, et al. The short Synacthen and insulin stress tests in the assessment of the hypothalamic-pituitary-adrenal axis. Clinical Endocrinology. 1996; 44: 141-6.[br][br]Nothing to Disclose: LEH, GK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 409 193 1496 SUN-471 PO45-02 Sunday 1293 2012


1290 ENDO12L_SUN-472 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Primary Adrenal Insufficiency and Testicular Tuberculosis: A Case Report Ane Caroline The Bonifacio Freire, Leticia Iervolino, Jose Viana Lima Junior, Nilza Scalissi, Tomas Zechini Barrese, Marilia Germano de Castro Irmandade Santa Casa de Miseric[oacute]rdia de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Irmandade Santa Csa de Miseric[oacute]rdia de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Introduction: Tuberculosis remains a major cause of primary adrenal insufficiency in many areas of the world, particularly in developing countries. Therefore, this differential diagnosis should always be considered in this clinical context, as the early identification and treatment may enable recovery of the adrenal function. (1)[br]Clinical Case: A 47 year-old man presented with progressive skin darkening, dizziness, postural hypotension and weight loss in the former year. During primary investigation, serum cortisol at 8 am was low (1,5 mcg/dL), adrenocorticotropic hormone level was very high (967 pg/mL) and an abdomen CT showed an enlarged right adrenal gland with 2 cystic areas and calcification foci. The patient was also diagnosed with primary hypothyroidism (TSH= 17,6 [micro]U/mL and fT4= 0,82 ng/dL). He had already initiated treatment with Levothyroxine 100 mcg and Prednisone 10 mg 9 months earlier, when he first arrived in our Endocrinology department. Further investigation, revealed a low serum sodium (131 mEq/L), high serum potassium (5,6 mEq/L), high serum renin (20,5 ng/mL/h) and low aldosterone (2,3 ng/dL). Mineralocorticoid replacement was initiated with Fludrocortisone 0,5 mg and Prednisone dose was reduced to 5mg per day. He had a negative tuberculin skin test, was HIV negative and an adrenal CT protocol scan showing nodular thickening of the adrenal glands, with a small calcification in the right adrenal. As a CT-guided biopsy was not feasible and a chest CT showed an image suggestive of primary complex of tuberculosis, empiric anti-tuberculosis treatment was indicated. However, subsequently, the patient complained about a hardened nodule on the left testicle, whose ultrasonography revealed a 2 cm nodule, with small cystic areas and slightly increased vascularization, suspicious for malignancy. He underwent orchiectomy and the pathology examination showed granulomatous reaction focally positive for acid-fast bacilli. He initiated, then, anti-tuberculous regimen and is being followed-up.[br]Conclusion: In a patient from a country where tuberculosis is endemic, this infection should always be considered in the differential diagnosis of a primary adrenal insufficiency, especially in association with enlarged or calcified adrenal glands. Extra-adrenal tuberculous involvement should be actively searched because it may provide indirect microbiologic or histologic clues. Besides pulmonary involvement, the genitourinary system should be carefully surveyed.[br][br](1) Al-Mamari A et al., Sultan Qaboos Univ Med J 2009; 9:324. (2) Kon YC et al., Endocrine practice 2002; 8:365.[br][br]Nothing to Disclose: ACTBF, LI, JVLJ, NS, TZB, MGdC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2033 193 1497 SUN-472 PO45-02 Sunday 1294 2012


1291 ENDO12L_SUN-473 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Primary Adrenal Burkitt-Like Lymphoma Presenting as Cushing Syndrome Ming Li, M Luiza Caramori University of Minnesota, Minneapolis, MN Introduction: Primary adrenal lymphoma is a rare malignancy of adrenal gland which is in its nature hormonally inactive.[br]Clinical Case: A 30 year old male with history of poorly controlled diabetes and hypertension presenting to the ED with abrupt onset of left upper quadrant abdominal pain. A non-contrast CT identified a 6 cm left adrenal mass. Hormonal workup was notable for elevated 24 hour urinary free cortisol (167 [micro]g/24 hr, normal [lt] 60 [micro]g/24 hr), and low DHEA-S level (76 [micro]g/dl, normal 86-560 [micro]g/dl). Physical exam was noted for a BMI of 31.7, buffalo hump and moon[apos]s face, even though other common features of Cushing[apos]s syndrome, such as purple striae and easy bruising, were lacking. Left adrenalectomy was performed, and the surgical pathology reported primary adrenal lymphoma. Further FISH, immunohistochemical, and flow cytometry analysis gave the eventual diagnosis of B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and Burkitt[apos]s lymphoma. Remarkably, 24 hour urinary free cortisol normalized 2 weeks after surgery (59.5 [micro]g/24 hr), and an overnight 1 mg dexamethasone suppression test showed suppression of cortisol levels (1.6 [micro]g/dl, normal [lt] 1.8 [micro]g/dl). The patient was treated with full course of R-HyperCVAD chemotherapy and the body PET/CT performed after completion of therapy showed no abnormal uptake.[br]Clinical Lesson: Even though a formal diagnosis of Cushing[apos]s syndrome was not fully established prior to surgery, the patient did carry many of its clinical features. We speculate that the patient could either have pseudo-Cushing[apos]s syndrome on presentation from his acute stress or obesity, or his lymphoma might have been producing an unknown substance stimulating adrenal production of cortisol. This case illustrates the need for complete evaluation of the hypothalamus-pituitary-adrenal axis prior to surgery, in cases with similar presentation.[br][br]Nothing to Disclose: ML, MLC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2053 193 1498 SUN-473 PO45-02 Sunday 1295 2012


1292 ENDO12L_SUN-474 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) X-Linked Adrenal Hypoplasia Congenita [mdash] A Novel Missense Mutation Stephanie Therese Chung, Carolyn Haekyung Chi, George Said Jeha, Morey William Haymond Baylor College of Medicine, Houston, TX Introduction: X-linked adrenal hypoplasia congenita (AHC), a rare cause of primary adrenal insufficiency, results from a loss of function of the [italic]NROB1[/italic] gene on Xp21.3. [italic]NROB1[/italic] encodes for the DAX-1 protein which is expressed in the urogenital ridge and fetal adrenal and is hypothesized to be instrumental in the development and function of the adrenal gland. Most [italic]NROB1[/italic] mutations are frameshift or nonsense mutations which provide limited information about the function of the various DAX-1 domains. Missense mutations offer some insight into the function of DAX-1 protein but comprise less than 20% of [italic]NROB1[/italic] mutations.[br]Clinical Case: A 12 day-old male presented with recurrent vomiting, weight loss, jaundice and hypotonia. On examination, he had generalized hyperpigmentation with normal blood pressure and male genitalia. Laboratory investigations revealed a normal CAH newborn screen, hyponatremic dehydration, mild hyperkalemia and unconjugated hyperbilirubinemia. Primary adrenal insufficiency was confirmed by a low baseline cortisol (2.9 [mu]g/dl, n [gt]8 [mu]g/dl), elevated adrenocorticotropic hormone (1761 pg/ml, n=6-48 pg/ml), elevated plasma renin activity (53.4 ng/ml/hr, n=2-35 ng/ml/hr), and an undetectable aldosterone ([lt]5 ng/dl). The 11-desoxycortisol of 1092ng/dl (n = 10-156 ng/dl) was the only elevated steroid precursor and there was no response to intraveneous 250[mu]g Cosyntropin. The 11-desoxycortisol suppressed completely after two months of glucocorticoid and mineralocorticoid treatment. Chromosomal microarray showed copy number loss of chromosome 10q21.1 with intact Xp22-p21 region. There are no reported associations between deletions on chromosome 10 and adrenal insufficiency. DNA [italic]NROB1[/italic] sequencing detected a nucleotide change of c.1094T[gt]C which would result in the change p.Leu365Pro not previously described or cited as a polymorphism. This highly conserved residue is within the second of three known mutation clusters in the critical ligand-binding domain thought to be essential to the DAX-1 function of transcriptional repression and in this clinical setting, this novel mutation is probably pathological.[br]Clinical Lessons: AHC can be associated with increased 11-desoxycortisol concentrations and this could represent remnant adrenal tissue rather than 11-[beta]hydroxlase deficiency in boys. [italic]NROB1[/italic] missense mutations can result in severe disease and provide further insights into the function of this elusive nuclear receptor protein in adrenal gland development.[br][br]Nothing to Disclose: STC, CHC, GSJ, MWH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 871 193 1499 SUN-474 PO45-02 Sunday 1296 2012


1293 ENDO12L_SUN-475 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Delayed Diagnosis of Addison[apos]s after 3 Unnecessary Surgical Procedures Aswathia Srinath, Natasha Mansell, Abbi Lulsegged Kings College Hospital, London, UK; Brighton and Sussex University Hospitals, Brighton, UK; South London Healthcare NHS Trust, Orpington, UK Primary adrenal insufficiency is a rare condition with an incidence of 0.8/100000 presents with diverse non specific clinical and biochemical features making diagnosis challenging which can be easily missed unless suspected.[br]A previously healthy 19y.o female presented with severe abdominal pain and vomiting. She had long standing abdominal discomfort and 4 days before her presentation she had laparoscopic gynaecological evaluation which was unremarkable.[br]Pulse rate 120/minute, BP 93/49, she had cold peripheries [amp] appeared peritonitic. Systemic examination showed no abnormality. Labs: Sodium 129mmol/L (135-145), CRP 63 ([lt] 10) [amp] she had metabolic acidosis: pH 7.25 [amp] bicarbonate 15.2mmolL.[br]She was thought to have abdominal sepsis and was give Intravenous fluid and antibiotics. CT Abdomen showed free fluid in peritoneum. Emergency laparotomy showed no significant abnormality. Postoperatively she needed ITU support including ventilation [amp] inotropes. Her condition further deteriorated [amp] underwent second laparotomy which was again unremarkable. Repeat CT Chest/Abdomen showed bilateral consolidation with pleural effusion.[br]Endocrine evaluation was carried out because of Hyponatremia: TSH 15.18pmol/L (0.5-5) fT4 9.1mmol/L (9-15) [amp] cortisol 24mmol/L (300-350).[br]She responded well to parenteral glucocorticoids. Synacten test showed cortisol levels 26nmol/L at 0 minutes, 42nmol/L at 30 minutes and 36nmol/L at 60 minutes. She made a remarkable recovery on steroid replacement and discharged with endocrine follow-up[br]Conclusion-Primary adrenal insufficiency is a life threatening endocrine disorder commonly missed. We report a case of a young lady who underwent 3 un-necessary operations and 2 Abdominal CT scans (equivalent to 800 chest X-rays) before the correct diagnosis of Addison[apos]s disease was made. A high index of suspicion is required in hyponatremic patients with unexplained abdominal pain. Once suspected the diagnosis is easy to exclude.[br][br]N Engl J Med 1996; 334:1403-1405.[br][br]Nothing to Disclose: AS, NM, AL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1227 193 1500 SUN-475 PO45-02 Sunday 1297 2012


1294 ENDO12L_SUN-476 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Systemic Amyloidosis Secondary to Familial Mediterranean Fever Presenting as Polyglandular Endocrine Failure Nicole M Tyer, Anagh Vora, Bhuta Sunita, Smooke-Praw Stephanie UCLA Medical Center, Los Angeles, CA; UCLA Medical Center, Los Angeles, CA [bold][underline]Introduction/Background[/underline][/bold][br]We present a patient with polyglandular endocrine failure due to systemic amyloidosis secondary to Familial Mediterranean Fever (FMF).[br][bold][underline]Clinical Case[/underline][/bold][br]A 41 year-old Armenian male status post kidney transplantation for idiopathic renal failure 8 years prior presented to the emergency room with abdominal pain, diarrhea, fever, hypotension, and acute kidney injury.[br]Infectious work-up was unrevealing and he remained hypotensive despite large volume fluid resuscitation. Given the history of chronic prednisone use for immunosuppression, an investigation for secondary adrenal insufficiency was completed and confirmed with a baseline morning cortisol of 6 mcg/dL that increased to 7 mcg/dL following administration of 250 micrograms of cosyntropin. Stress dose glucocorticoid therapy improved the blood pressure, but the abdominal complaints and acute kidney injury persisted.[br]Medical history was also significant for post-surgical hypothyroidism, and a careful chart review was performed. Thyroid pathology from ten years ago revealed amyloid goiter; no further work-up was pursued at that time.[br]In light of the amyloid goiter, idiopathic renal failure, and new onset kidney failure in the transplanted kidney, a diagnosis of Systemic Amyloidosis was considered. A biopsy of the transplanted kidney was obtained and indeed revealed amyloid infiltration.[br]On further questioning, the patient described similar [ldquo]attacks[rdquo] of abdominal pain, fever, and diarrhea that have occurred 3 to 4 times per year since childhood. Ibuprofen often alleviated these symptoms; other family members also experience these attacks. Given the patient[apos]s Armenian background, recurrent abdominal attacks, and systemic amyloidosis, a unifying diagnosis of FMF was considered (1).[br]Genetic analysis of the FMF gene revealed compound heterozygosity at two alleles (M680I G/C and M680I G/A), consistent with a diagnosis of FMF. Gastroenterology was consulted; colchicine was commenced with marked improvement in symptoms and frequency of attacks. Genetic counseling has been recommended for his 15 year-old daughter and other family members.[br][bold][underline]Clinical Lessons/Conclusion[/underline][/bold][br]Untreated FMF can lead to amyloid deposition in the kidney, liver, spleen, and gut, as well as endocrine organs, including pituitary gland, thyroid, parathyroid, adrenal, and testes (2). Clinicians should maintain a high index of suspicion for this diagnosis as treatment can prevent multi-organ failure.[br][br](1) Danovitch G et al. Clinical Endocrinology 1979; 6:595. (2) Keven K et al. Am J Kid Dis 2001; 6:E39.[br][br]Nothing to Disclose: NMT, AV, BS, S-PS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1747 193 1501 SUN-476 PO45-02 Sunday 1298 2012


1295 ENDO12L_SUN-477 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Acute Diabetes Insipidus Associated with Placental Abruption Mediated by Release of Vasopressinase into the Maternal Bloodstream Aigerim Bizhanova, Amisha Wallia, Wenyu Huang, Susan Goldsmith, Dana Gossett, Peter Kopp Northwestern University, Feinberg School of Medicine, Chicago, IL; Northwestern University, Feinberg School of Medicine, Chicago, IL Transient diabetes insipidus (DI) is a rare complication of pregnancy characterized by polyuria, polydypsia, and excessive thirst. This form of DI usually occurs during late pregnancy and/or the immediate postpartum period and is due to increased systemic levels of vasopressinase, a placental enzyme that degrades arginine-vasopressin (AVP) but not 1-deamino-8-D-arginine vasopressin (DDAVP), a synthetic analogue of AVP.[br]We report a patient (G3P0) who had a twin pregnancy and then presented at week 33 with placental abruption prompting a Cesarean section. Within a few hours post surgery, she developed acute severe DI. Her other pituitary axes were intact. A pituitary MRI showed a slightly prominent pituitary gland but no signs of infarction or hypophysitis. The DI could be easily controlled with DDAVP but not with AVP. Six weeks postpartum, DDAVP was gradually weaned off. Off DDAVP, her oral intake and urinary output were normal. Prior to her pregnancy, she never noticed polyuria or polydipsia. Moreover, the patient did not develop any signs and symptoms suggestive for recurrence of DI during a second uncomplicated pregnancy and delivery.[br]In order to evaluate whether the acute onset of DI could be associated with the release of vasopressinase secondary to the placental abruption, sera collected during the postpartum period and at 5 months were tested for vasopressinase activity. TSA-201 cells were transfected with an AVPR2 cDNA and a cAMP-reponsive luciferase reporter construct. Cells were treated with AVP or DDAVP in absence and presence of the patient sera. In the absence of serum, both AVP and DDAVP induced a [sim]50-fold activation of the reporter construct. In contrast, the AVP-induced activation was completely abolished in cells treated with the serum from the postpartum period, while the response to DDAVP was unaltered. The serum collected at 5 month did not reduce the AVP-mediated induction.[br]In conclusion, the clinical course and the laboratory results suggest that large amounts of vasopressinase were acutely released into the maternal blood stream secondary to the abruption. Vasopressinase degrades endogenous and exogenous AVP, but not DDAVP. With clearance of the vasopressinase activity, the polyuria disappeared and the patient did not have any signs suggestive of DI. The observation indicates that placental abruption can rarely be associated with acute transient DI that is mediated by the release of vasopressinase into the maternal blood stream.[br][br]Nothing to Disclose: AB, AW, WH, SG, DG, PK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2234 193 1502 SUN-477 PO45-02 Sunday 1299 2012


1296 ENDO12L_SUN-478 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) The Stubborn Puzzle: A Case Report of Atypical Type 2 (Type 4) Autoimmune Polyglandular Syndrome (APS-2) Combination with Common Variable Immunodeficiency (CVID) Ieva Ruza, Inta Leitane Riga East Clinical University Hospital, Riga, Latvia; Tornakalna Outpatient Clinic, Riga, Latvia [bold]Introduction[/bold]: Wide spectrum of different autoimmune polyglandular syndromes (APS) are described, now all divided into 2 subtypes (1). We report an atypical combination of endocrine autoimmunity with common variable immunodeficiency (CVID).[br][bold]Report[/bold]: A 50-years old white woman was referred to our clinic for an evaluation due to suspected adrenal insufficiency. 2 months earlier during examination her GP had discovered hyperpigmentation of her back and hips, only skin biopsy was done (possible Addison[apos]s disease?). When severe fatigue after mild viral infection evolved, she was referred to endocrinologist.[br]First examination revealed also dark face and palms, BP 85/60 mmHg, no orthostatic changes. She had a childhood history of frequent respiratory infections, failure to thrive, underweight, abdominal bloating, diarrhea exacerbated by fatty food, no lactose intolerance. Due to gastrointestinal symptoms she was investigated 8 and 3 years ago; atrophic gastritis, duodenal reactive lymphoid hyperplasia, CVID was diagnosed, malignancy, systemic disease or TB excluded. Progressive osteoporosis was diagnosed 3 years ago, bisphosphonates and calcium started. She was a virgin, menopausal for 3 years. Family history showed no features to support possible hereditary disease.[br]Biochemical tests demonstrated elevated ACTH, low serum and 24h urinary cortisol, marked hypocalcemia, low 24h urinary calcium, low calcidiol, elevated serum PTH. FSH, LH, TSH, free thyroxin was normal, vitamin B[sub]12[/sub] decreased, no data on diabetes mellitus. Antibody screen was positive for ovaries, but negative for adrenals, thyroid peroxidase and transglutaminase. No substantial changes seen in other tests. There were no pituitary changes on head CT.[br]Diagnose of primary adrenal insufficiency, hypocalcemia, secondary hyperparathyroidism, atrophic gastritis, intestinal malabsorbtion, secondary osteoporosis and CVID was done. Based on all findings, we suggest the final diagnose of type 2 (type 4) autoimmune polyglandular syndrome (APS-2), combined with CVID and possible undiagnosed celiac disease.[br]Treatment was started with hydrocortisone and calcitriol, calcium dosage increased, bisphosphonates continued. A substantial improvement of symptoms was seen in a control visit 6 months later. We would like to stress the importance of screening for underlying autoimmune diseases in case of diagnosed endocrine autoimmune pathology.[br][br](1) Kahaly GJ, European Journal of Endocrinology 2009; 161:11-20.[br][br]Nothing to Disclose: IR, IL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1887 193 1503 SUN-478 PO45-02 Sunday 1300 2012


1297 ENDO12L_SUN-479 POSTER SESSION: Adrenal Disorders Case Reports II (1:30 PM-3:30 PM) Paraneoplastic Syndrome of Inappropriate Antidiuretic Hormone and Superior Vena Cava Syndrome in a Pulmonary Microcytoma Gian Pio Sorice, Caterina Policola, Teresa Mezza, Giovanna Muscogiuri, Lorenzo Di Pizio, Vin Sin Alice Sun, Barbara Altieri, Maria Chiara Fabiano, Andrea Giaccari, Silvia Della Casa Universit[agrave] Cattolica del Sacro Cuore, Roma, Italy It is estimated that 10% of patients with small-cell lung cancer (SCLC) have Syndrome of Inappropriate Secretion of Antidiuretic Hormone (SIADH). We report a case of a SCLC presented with SIADH and superior vena cava obstruction. A 67-year-old woman was admitted because of hyponatriemia, referred to the emergency department complaining headache, nausea, dizziness. Laboratory studies performed at the admission revealed serum sodium level 112mmol/L, potassium 4.4mmol/L, and plasma osmolality 275mmol. No impairment of pituitary, thyroid and adrenal function was detected. She was treated with 3% hypertonic saline. Chest X-ray revealed a multilobed opacity in the upper right anterior mediastinal; CT-scan showed a solid tissue (3x2 cm) localized in the right upper lobe; this nodule was in continuity with an infiltrative solid tissue of the mediastinal paratracheal right and ipsilateral hilum (7x8x6 cm). This tissue compressed the distal portion of the trachea, displaced the superior vena cava, encompassed the arch of azygos vein and the superior pulmonary vein. Three days after the admission, patient started to complain chest tightness and cough. On physical examination, facial edema and venous distension in the neck were detectable. After another 1 week, cyanosis of the lips and facial plethora developed. The biopsy demonstrated SCLC, with positivity for CD56, synaptophysin, chromogranin A, Ki67 index by about 70%. NSE and Cyfra 21.1 were increased, 53.2 ([lt]16.5) and 20.4 ng/ml ([lt]3.30), respectively. ADH was undetectable. The patient underwent the placement of a self-expanding endovascular stent, with partial remission of symptoms. At meantime, the correction of hyponatremia required also treatment with a competitive vasopressin receptor-2 antagonist, discontinued when patient underwent chemiotherapy. After three months, the solid tissue was reduced, symptoms restored, serum sodium and markers levels normalized. SIADH is characterized by euvolemic hyponatremia, low plasma osmolality, high urinary osmolality, lack of evidence of other hyponatremic diseases. This clinical case confirms euvolemic hyponatremia may be a sign of paraneoplastic syndrome, as a manifestation of SIADH even with negative laboratory test for ADH; inversely, the superior vena cava syndrome is an uncommon manifestation of SCLC, worsening prognosis and QOL of patients. Vasopressin receptor-2 antagonists may be useful in restoring normal level of serum sodium during treatment of main disease.[br][br]Nothing to Disclose: GPS, CP, TM, GM, LDP, VSAS, BA, MCF, AG, SDC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 984 193 1504 SUN-479 PO45-02 Sunday 1301 2012


1298 ENDO12L_SUN-480 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Patient and Disease Characteristics in a Single-Center Retrospective Study of 360 Adrenocortical Carcinoma (ACC) Patients Tobias Else, Andrew Williams, Gary Hammer University of Michigan, Ann Arbor, MI; University of Michigan, Ann Arbor, MI Introduction: Adrenocortical carcinoma is a rare endocrine malignancy and even basic epidemiological data are not well established. In order to define the basic patient and disease characteristics we analyzed 360 ACC patients. [br]Methods: Single center retrospective analysis of ACC patients in a large academic tertiary referral center with specialized endocrine oncology service. ACC patients were identified in electronic medical record covering at least a 13 year time frame (1998 [ndash] 2011). Medical records were reviewed for patient and disease characteristics (gender, race, age at diagnosis, stage at diagnosis, hormone production, laterality, and diagnosis during pregnancy). Basic data were included in a Cox regression model to define hazard ratios for factors impacting survival.[br]Results: The median age at diagnosis was 47yrs. Female to male ratio was 1.5. 153 (43%) ACC were non-functional and hormone production (clinical or biochemical) was found in 207 (57%) (cortisol 38%, androgens 18%, mineralocorticoid 5%, estradiol 2%, multiple hormones 37%). Reason for initial imaging and diagnostic work up was known for 345 patients. Hormone excess was the most common cause to initiate evaluation (38%), followed by abdominal or flank pain (36%). Another 9% patients were evaluated for unspecific, but tumor related symptoms. 17% of ACC were diagnosed by imaging conducted for reasons unrelated to ACC. The stage at diagnoses was stage 1 (4%), stage 2 (45%), stage 3 (26%) and stage 4 (25%). In a Cox regression model stage 4 at diagnosis (HR=5.35, CI(95%) 2.15-13.34), age at diagnosis (HR=1.02, CI(95%) 1-1.03) and cortisol secretion (HR=1.65, CI(95%) 1.25-2.18) significantly affected survival.[br]Conclusion: These data confirm reported epidemiological characteristics and identify advanced stage disease, age and cortisol production as independent risk factors.[br][br]Sources of Research Support: NIH grant T32-DK007245.[br][br]Nothing to Disclose: TE, AW, GH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2232 194 1505 SUN-480 PO01-01 Sunday 1302 2012


1299 ENDO12L_SUN-481 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Clinical Characteristics of Adrenocortical Carcinomas Less Than 4 cm in Size: A Case Series Aashish Samat, Amir H Hamrahian, Khaled M Elsayes, Erick M Remer, Priya Rao, Amr Fergany, Eren Berber, Mouhammed A Habra Cleveland Clinic, Cleveland, OH; The University of Texas MD Anderson Cancer Center, Houston, TX; Cleveland Clinic, Cleveland, OH; The University of Texas MD Anderson Cancer Center, Houston, TX; Cleveland Clinic, Cleveland, OH Introduction: Adrenocortical carcinoma (ACC) is a rare endocrine malignancy with high mortality. Tumor size is an important determinate of the stage of disease. Most guidelines recommend resection of adrenal masses [gt] 4 cm in size. We describe common clinical features of adult patients with histologically proven ACC [le] 4 cm in size.[br]Methods: Retrospective Chart review at MD Anderson Cancer Center and Cleveland Clinic (1998-2010). The histological diagnosis of all patients was reconfirmed by an experienced pathologist.[br]Results: We identified 9 patients with ACC: 3 presented as an incidentaloma, 3 had Cushing[apos]s syndrome, 2 had aldosterone hypersecretion, and 1 presented with flank pain. The median (range) age was 57 (34-77) years and F/M sex ratio was 5/4. Despite small tumor size, one patient had stage 3 and three had stage 4 disease at diagnosis. Two patients had a history of another primary malignancy and five had a history of malignancy in a first degree relative. CT scans of four patients were available for review. The most common CT features were: ill defined tumor or irregular border in 3, heterogeneity in 2, and necrosis in 1 tumor. The median tumor size on CT and pathology were 2.0 cm (1.3-3.2) and 3.6 cm (1-4), respectively. Noncontrast CT attenuation values were available in 2 patients and were 35 and 41 HU. One patient with metastatic disease received systemic chemotherapy while 8 patients underwent surgery, of which 4 had an isolated tumor with clear surgical margins. None was treated with mitotane postoperatively. The patients were followed for 14.9 months (1-49). Six patients died from ACC (cause of death unknown in one) including 2 patients with an isolated tumor and clear surgical margins.[br]Discussion: We found a high rate of mortality in patients with small ACC including a 75% overall mortality rate in those with an isolated [le] 4 cm tumor at the time of diagnosis with a clear surgical margin. It is unknown if the lack of usage of mitotane had an effect on mortality. Referral bias could be a major factor in the increased mortality in these cases. Among patients with available images for review, all had some features that are not typical of benign nodules.[br]Conclusion: Small ACCs carry a high mortality rate. Close follow up or additional studies need to be considered when confronting an adrenal mass [le] 4 cm in size with worrisome radiological features such as a noncontrast CT attenuation [gt] 30 HU, heterogeneity, necrosis or an irregular border.[br][br]Nothing to Disclose: AS, AHH, KME, EMR, PR, AF, EB, MAH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1870 194 1506 SUN-481 PO01-01 Sunday 1303 2012


1300 ENDO12L_SUN-482 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Clinical Characteristics and Follow-Up Aspects of 287 Korean Patients with Adrenal Incidentaloma Yoon Young Cho, Hye Jeong Kim, Se Won Kim, Sun-Mi Park, Ji Cheol Bae, Sun Wook Kim, Jae Hoon Chung, Jae Hyeon Kim Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea Adrenal incidentalomas are defined as incidentally discovered adrenal mass lesion [ge] 1cm in diameter without symptoms suggesting adrenal disease. The aim of this study is to investigate the clinical characteristics and follow-up findings of subjects with adrenal incidentalomas in single tertiary hospital in South Korea.[br]We retrospectively studied 287 adrenal incidentaloma subjects who underwent radiologic and endocrinologic evaluation at Samsung Medical Center, Seoul, South Korea between January 2004 and July 2011. To our knowledge, this study is the largest adrenal incidentalomas data for in Asian population.[br]Among 287 subjects, 173 (60.3%) subjects were men and most (87.5%) of participants proved to have non-functioning tumors. Functioning tumors were seen in 36 (12.5%) subjects, there were 22 (7.6%) subjects with subclinical Cushing syndrome (sCS), 9 (3.1%) subjects with pheochromocytoma and 5 (1.7%) subjects with primary hyperaldosteronism. Malignant adrenal tumors were discovered in 3 cases, 2 (0.7%) patients were primary adrenal cancer and 1 (0.3%) subject was adrenal metastasis from lung cancer. Significant risk factors for functioning tumor were female gender, lower BMI and large adrenal mass. During follow-up (a mean of 23 months) of 146 patients, 2 (1.4%) subjects developed hormonal changes to functioning tumors. One was diagnosed with sCS and medically followed up. The other was confirmed as pheochromocytoma by histopathology. No malignant transformation was found in these subjects. Initial hormonal and radiological evaluation for adrenal incidentaloma is more important than follow-up tests because functional or malignant changes are very rare.[br][br]Nothing to Disclose: YYC, HJK, SWK, S-MP, JCB, SWK, JHC, JHK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1172 194 1507 SUN-482 PO01-01 Sunday 1304 2012


1301 ENDO12L_SUN-483 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) What Is Appropriate Diagnostic Follow-Up for Patients with Adrenal Incidentalomas and a Negative Initial Work-Up for Primary Aldosteronism? Ekaterina Rogal, Dmitrii Beltsevich, Valentin Fadeev, Nataliya Molashenko, Ekaterina Pigarova, Nikolai Kyznetsov Endocrinologic Research Center (ERC), Moscow, Russian Federation Background: The optimal frequency and duration of follow-up of patients with adrenal incidentalomas is uncertain. Serial imaging in addition to annual hormonal evaluation of glucocorticoids and catecholamines is commonly recommended for an average of 4 years. However, there is little evidence for repeating hormonal testing of autonomous aldosterone hypersecretion.[br]Clinical case: Bilateral adrenal masses, measuring 1.2 and 1.5 cm, were found incidentally by CT in a 35-year woman. Her medical history was remarkable only for hypertension, poorly controlled with 4 antihypertensive medications. Initial hormonal testing - Metanephrine 163 mcg (n 25-312 mcg/24h), Normetanephrine 368 mcg (n 35-445 mcg/24h) in 24-hour urine sample, serum cortisol after overnight 1mg dexamethasone suppression test 1.6 [mu]g/dL (n[lt]1,8 [mu]g/dL), serum aldosterone 18 ng/dL (n 1-10,5 ng/dL), PRA 0.1 ng/mL/h (n 0,5-1,9 ng/mL/h) and plasma aldosterone after saline infusion test 4.5 ng/dL (n[lt]5 ng/dL). On one year follow up, the size and appearance of the adrenal masses on CT were unchanged. Urine fractionated metanephrines and 1 mg overnight dexamethasone test were normal, however plasma aldosterone after saline infusion test was borderline at 8.2 ng/dL (borderline 5-10 ng/dL). Given the patient[apos]s early onset of essential hypertension prior to age 30 and poor control of hypertension despite multiple medications adrenal venous sampling (AVS) was performed without cosyntropin stimulation. Right adrenal vein [ndash] aldosterone 746 ng/dL, cortisol[gt]631 [mu]g/dL; left adrenal vein aldosterone 610 ng/dL, cortisol[gt]631 [mu]g/dL, IVC - aldosterone 164 ng/dL, cortisol 42 [mu]g/dL. Bilateral cortisol adrenal vein values were unable to be calculated accurately despite maximum possible serum[apos]s dilution (more than 10 times) and high probability of laboratory error in this setting. A second attempt was made with similarly ambiguous cortisol and now aldosterone results as follows: right adrenal vein [ndash] aldosterone[gt]1354 ng/dL, cortisol[gt]631 [mu]g/dL; left adrenal vein aldosterone[gt]1354 ng/dL, cortisol[gt]631 [mu]g/dL, IVC - aldosterone 164 ng/dL, cortisol 42 [mu]g/dL. Without definitive results, the patient was continued on her previous medications with the addition of eplerenone. Blood pressure at a six month follow-up was 150/90 mm Hg.[br]Conclusion: This clinical case demonstrates the necessity of close follow-up of patients with adrenal incidentalomas and negative initial testing of aldosterone hypersecretion.[br][br]Nothing to Disclose: ER, DB, VF, NM, EP, NK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 369 194 1508 SUN-483 PO01-01 Sunday 1305 2012


1302 ENDO12L_SUN-484 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Malignancy Predictors of Adrenal Incidentalomas Vered Seri, Petachia Reissman, Herbert R Freund, Liat Appelbaum, Gil Leibowitz, Merav Fraenkel Hadassah-Hebrew University Medical Center, Jerusalem, Israel; Shaare-Zedek Medical Center, Jerusalem, Israel; Hadassah, Hebrew University Medical Center, Mount Scopus, Jerusalem, Israel; Hadassah-Hebrew University Medical Center, Jerusalem, Israel Background:[br]Adrenal incidentalomas are common on imaging studies and should be assessed for hormone secretion and risk for malignancy.[br]Objective:[br]The aim of this study was to identify the adrenal incidentaloma characteristics which predict malignancy.[br]Methods: We performed a retrospective study of all adrenalectomies performed in three academic hospitals in Jerusalem between1999-2008. Hormone secretion and imaging characteristics were analyzed. The prevalence of each variable and its correlation with the final diagnosis were studied.[br]Results:[br]Two-hundred thirty five patients were studied. Sixty percent of the patients were women and the mean age was 52 years. In 28.9% of the patients, the lesion was an incidental finding; and mean tumor diameter was 4.8 cm. Hormonal hypersecretion was found in 67.8%.[br]12.8% of all adrenal lesions were malignant: 43% of which were adrenocortical carcinomas and 57% were metastasis. Sixty-nine percent of all adrenal lesions were benign and 16.6% were pheochromacytomas.[br]A multiple logistic regression analysis showed that tumor size correlated with the risk for malignancy (p=0.001), and that hormone secretion was associated with a lower risk for malignancy (p=0.05). A ROC analysis showed that tumor diameter of 4.6 cm was the optimal cut-off size for differentiating between benign and malignant tumors with a sensitivity of 77% and specificity of 69%.[br]The incidence of malignancy in patients that were operated due to imaging findings suggestive of malignancy was 17.6%.[br]Conclusions:[br]In our cohort, lesion size on CT imaging was the most powerful predictor of malignancy, while hormonal hypersecretion was associated with a final diagnosis of a benign tumor. Most lesions suspected to be malignant based on imaging studies were benign. Despite of the infrequency of malignancy under these circumstances, adrenalectomy is probably recommended to allow an early diagnosis of rare adrenocortical carcinomas.[br][br]Nothing to Disclose: VS, PR, HRF, LA, GL, MF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 365 194 1509 SUN-484 PO01-01 Sunday 1306 2012


1303 ENDO12L_SUN-485 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) E-Selectin Levels and Insulin Resistance in Patients with Nonfunctional Adrenal Incidentalomas Basak Karbek, Oya Topaloglu, Evrim Cakir, Nujen Colak, Muyesser Aslan, Askin Gungunes, Ilknur Unsal, Bekir Ucan, Taner Demirci, Melia Karakose, Tuncay Delibasi Diskapi Yildirim Beyazit Teachin and Research Hospital, Ankara, Turkey Background: The association of nonfunctioning adrenal adenomas (NFA) with cardiovascular risk factors is well known but the mechanisms of this relationship are still subject of discussions.We aimed to investigate, circulating e-selectin levels in subjects with clinically silent adrenal adenomas and also to search for the association of e-selectin levels with insulin resistance(IR) and carotid artery intima-media thickness(CIMT) and cardiovascular risk factors.[br]Methods:A total of thirty-nine patients with NFA (27 women and 12 men mean age,55,61 1[plusmn] 0,73 years) and thirty four healthy comparison subjects (24 women and 10 men; mean age,51,52 [plusmn] 8,09 years) were enrolled.Serum levels of e-selectin were assayed by enzyme-linked immunosorbent assay in healty controls and subject with NFA and typical computed tomographic features of cortical adenoma, who were not affected by diabetes mellitus, hypertension, or other relevant diseases.In all subjects, we measured, in the fasting state, serum cortisol at 08:00 am, upright plasma aldosterone and plasma renin activity (PRA); urinary metanephrine, normetanephrine and free cortisol excretion were assessed on a 24-h sample.Dynamic tests included cortisol after 1-mg dexamethasone test; we performed overnight 2-mg dexamethasone test when patients did not display plasma cortisol suppression.Serum HsCRP, lipid profile, insulin levels and the homeostasis model assessment of insulin resistance (HOMA-IR) were evaluated.High-resolution B-mode ultrasonography was performed.[br]Results:The two study groups did not differ as to age, sex, BMI and blood pressure.Serum e-selectin levels were significantly elevated in patient with NFA when compared with that of control subjects (NFA:14,9 [plusmn] 4,76, control:12,2 [plusmn] 4.07 p[lt]0.01).CIMT and HsCRP levels were similar between two groups.Insulin resistance as calculated by HOMA-IR was higher in the NFA groups than that in the control group(p[lt]0.01).Serum e-selectin levels showed a statistically significant association with Hs-CRP(r=0,751, p[lt]0.001), HOMA-IR(r=0,575, p[lt]0.001) and CIMT(r=0,762, p[lt]0.001). No significant correlation was identified between e-selectin levels and age, gender, BMI and other other biochemical parameters.[br]Resuts:There was no difference between patient and control groups in other known cardiovascular risk factors, including age, sex, cholesterol panels, HT, and smoking.However, serum e-selectin levels in patients were high and there was a positive correlation with CIMT and HOMA-IR.[br][br]Nothing to Disclose: BK, OT, EC, NC, MA, AG, IU, BU, TD, MK, TD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 809 194 1510 SUN-485 PO01-01 Sunday 1307 2012


1304 ENDO12L_SUN-486 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Performances of Tomodensitometric Scannography for the Evaluation of Incidentalomas Patrice Jahanpur, Claire Auder, Delphine Gaye, Stephanie Reich, Marie-Laure Nunes, Muriel Cogne, Francois Favier, Alexandre Rault, Denis Collet, Tristan Wagner, Antoine Tabarin CHU Bordeaux, Pessac, France; CHU Bordeaux, Pessac, France; CHR, Saint Pierre, French Southern Territories; CHU Bordeaux, Pessac, France Background: CT scanning is the cornerstone evaluation of adrenal tumors. Low unenhanced attenuation values (UA) and intense wash-out of contrast media have been proposed to differentiate cortical adenomas from non adenomas with excellent performances. However, most studies have been performed in surgical or oncologic settings and no series has been conducted on adrenal incidentalomas recruited in an endocrine setting.[br]Objective: Assess the performance of CT-scanning to differentiate adenomas from non adenomas amongst adrenal incidentalomas.[br]Methods: Retrospective study in a single endocrinology department. 175 patients with 194 tumors were studied. Measurement of UA, relative percentage wash-out (RPW) and absolute percentage wash-out (APW) were performed in 165, 87 and 82 patients respectively. 82 incidentalomas were operated (35 adenomas, 16 adrenal cortical carcinomas (ACC), 23 pheochromocytomas (Ph), 1 metastasis and 7 miscellaneous histologic types).[br]The diagnosis of adenoma was proved on the basis of a stable lesion size or an attenuation value [lt]10 HU at nonenhanced follow-up CT for at least 6 months (mean 3,6 years; range 6 months[ndash]16 years).[br]112 non-operated tumors were considered as benign adenomas on stringent CT criteria including size [le]40mm and [le]20HU (Housfield Units) or no significative growth after 3,6 years of median follow-up or a UA lower than 10HU during follow-up.[br]158 CT scans were re-examined by 2 independent radiologists who were blinded to the final diagnosis.[br]Results: the median UA in Housfield Units (HU) for adenomas was 9,0[plusmn] 13,4, 38,0 [plusmn] 19,1 for ACC, 39,1 [plusmn] 13,7 for Ph and 28,2 [plusmn] 8,1 for other incidentalomas (p[lt]0,0001).[br]Using ROC analysis, sensibility and specificity were: 79,6% and 91,5% for a threshold of size of 30mm; 77,6% and 93,5% for a threshold of 20HU for UA.[br]Using ROC analysis, the performance of RPW were better than that of APW (p[lt]0,02) with sensibility of 80,6% and specificity of 95,0% using a threshold [gt]39%.[br]The combination of size 40mm and UA 20HU had 72,4% sensitivity and 100% specificity.[br]37 adenomas did not fulfill the size and UA criteria, 15 were correctly identified using the RPW.[br]The inter-rater agreement Kappa coefficient was good for size, UA and RPW (80-86%) but low for APW (66%).[br]Conclusions: the combination of size [le]40mm and UA [le]20HU is an accurate criterion for the diagnosis of adenomas amongst incidentalomas. A wash out procedure with RPW threshold [ge]40% increases the sensitivity in ambiguous cases.[br][br]Nothing to Disclose: PJ, CA, DG, SR, M-LN, MC, FF, AR, DC, TW, AT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1888 194 1511 SUN-486 PO01-01 Sunday 1308 2012


1305 ENDO12L_SUN-487 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Determining the SUV Value of the Adrenal Incidentalomas with Positron Emission Tomography Tugba Gumus, Mine Sencan Eren, Mehmet Calan, Ozhan Ozdogan, Sevinc Eraslan, Firat Bayraktar, Hatice Durak, Abdurrahman Comlekci Dokuz Eylul University Medical Faculty, Izmir, Turkey; Dokuz Eylul University Medical Faculty, Izmir, Turkey Purpose: To determine the average SUV value for benign adenomas. We evaluated the patients with benign and non-secreting adenomas retrospectively, who had FDG PET/CT to asses the adrenal mass and who were followed.[br]Material and method: This study was a retrospective and descriptive investigation. Patients who were followed for adrenal incidentaloma in our endocrinology department between 15.01.2010 and 15.10. 2010 were included. The patients with adrenal adenoma due to MRI and those hormonally inactive were included if they had PET/CT scan. To confirm the hormonal inactivity, plasma ACTH was measured for subclinical cushing syndrome and supression test with 1mg dexamethasone was performed.(DST). If the morning cortisol value was below 1.8 g/dl after 1 mg DST performance, patient was accepted as inactive for subclinic cushing. The urinary excretion of catecholamine metabolites (metanephrine, normetanephrine, vanil mandelic acid and homovalinic acid) were evaluated for phaeochromocytoma and the patients with normal results were included in the study. In hypertensive or hypokalemic patients plasma aldosterone concentration level and aldosterone/renin rate were evaluated in order to determine the primary aldosteronism. Cut off values were [lt] 15 ng/dL and [lt]20 for plasma aldosterone concentration and aldosteron/renin rate, respectively. The patients with lower values, were accepted hormonally inactive for primary aldosteronism.[br]Totally 33 patients and 44 adrenal adenomas were evaluated. Of the patients 10 were male and 23 were female. Average diameter for 44 adrenal adenomas was 2.35 cm(min:1.1, max:6,5cm) and average SUV was 2.48 (min: 1, max:4.5). Evalution of corelation between SUV values of adrenal adenomas and diameter sizes was a moderately strong positive and statistically significant relationship was found between them. (p:0.022, r:0.346).[br]Conclusion: FDG PET/CT is a valuable imaging method for diagnosis, staging and treatment response in oncology patients. Because it is widely used, diagnosis of adrenal masses with FDG uptake is increasing. Diagnosis of the adrenal masses in oncology patients is necessary for appropriate management and prognostic evaluation. However no SUV is helpfull to differentiate the malign and benign adrenal masses. In this study we evaluated the hormonally inactive adrenal incidentalomas with FDG PET/CT and determined the SUV cutoff as 2.48.[br][br]Nothing to Disclose: TG, MSE, MC, OO, SE, FB, HD, AC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 626 194 1512 SUN-487 PO01-01 Sunday 1309 2012


1306 ENDO12L_SUN-488 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Patients with Subclinical Cushing Syndrome Due to ACTH-Independent Macronodular Adrenal Hyperplasia (AIMAH) May Exhibit Adrenal Nodules with Increased 18F-FDG Uptake, Similarly to Adrenal Carcinomas and Metastases Guilherme Asmar Alencar, Berenice Bilharinho Mendonca, Antonio Marcondes Lerario, Lilian Yuri Itaya Yamaga, Maria Candida Barisson Villares Fragoso Hospital das Cl[iacute]nicas, Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Hospital Israelita Albert Einstein, S[atilde]o Paulo, Brazil Background: AIMAH is a benign adrenal disease and a rare cause of endogenous Cushing[apos]s syndrome. Integrated 2-[fluorine-18]-fluoro-2-deoxy-D-glucose positron emission tomography/computed tomography (18F-FDG-PET/CT) is a diagnostic method that has been used to discriminate benign from malignant adrenal masses (1). We recently demonstrated that patients with overt Cushing[apos]s syndrome due to AIMAH may exhibit an intense 18F-FDG uptake on a PET/CT, similarly to patients with adrenal carcinomas and metastases (2). Objective: The aim of this study was to investigate the pattern of 18F-FDG uptake in the first stages of AIMAH, when patients still present a subclinical Cushing[apos]s syndrome and small adrenal nodules. Methods: Two brothers (51 and 48 years old) belonging to a pedigree with familial AIMAH were diagnosed with subclinical Cushing[apos]s syndrome. Both exhibited an abnormal overnight 1-mg dexamethasone suppression test (plasma cortisol 5.7 and 3.1, respectively; normal, [lt]1.8 [mu]g/dL), with serum dexamethasone levels in the expected range (256 and 271 respectively; expected, 180-550 ng/dL). Midnight salivary cortisol and 24-hour urinary total cortisol were in the normal range and ACTH was in the lower level of the normal range (8 and 10, respectively; normal 5-46 pg/mL). Unenhanced computed tomography revealed adrenal macronodules, with attenuation values up to 14 and 16 Hounsfield units, respectively. The patients underwent a18F-FDG-PET/CT study. Results: In both patients, transaxial images of 18F-FDG-PET/CT revealed that some of the adrenal nodules presented an increased 18F-FDG uptake in comparison to liver uptake. The maximum standardized uptake value (SUVmax), which is an index used to assess disease activity in FDG-PET imaging, was really increased in some of these nodules, reaching levels usually seen in malignant tumors and metastases (SUVmax [gt]3.1). Conclusions: Even in the first stages of AIMAH, when the secretion of cortisol is mildly increased, it is already possible to demonstrate the presence of adrenal nodules with an intense 18F-FDG uptake. Therefore, we reinforce that AIMAH should be considered in the differential diagnosis of adrenal lesions with increased 18F-FDG activity, such as carcinomas and metastases.[br][br](1) Blake MA et al. American Journal of Roentgenology 2010; 194:1450-1460. (2) Alencar GA et al. J Clin Endocrinol Metab 2011; 96(1):3300-3301.[br][br]Sources of Research Support: Brazilian Federal Agency for Support and Evaluation of Graduate Education (CAPES 2590-11-5/PDSE); Sao Paulo Research Foundation (FAPESP 2010/12702-1).[br][br]Nothing to Disclose: GAA, BBM, AML, LYIY, MCBVF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 412 194 1513 SUN-488 PO01-01 Sunday 1310 2012


1307 ENDO12L_SUN-489 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) The Value of Adrenal Venous Sampling for Lesion Localization in Primary Aldosteronism Hyung Jin Choi, Yoon Ji Kim, Eun Mee Oh, Jung Hee Kim, Kyu Eun Lee, Seong Yeon Kim Seoul National University College of Medicine, Seoul, Korea; Seoul National University College of Medicine, Seoul, Korea Context. Distinguishing between unilateral and bilateral adrenal hypersecretion is mandatory for surgical treatment of primary aldosteronism (PA). Adrenal venous sampling (AVS) is considered the gold standard for identification and localization of the responsible lesion.[br]Objective. The objective of this study was to determine the usefulness of AVS in PA patients.[br]Design. From January, 2001 to October, 2011, 86 patients with the biochemical diagnosis of PA were retrospectively analyzed.[br]Setting. The study was performed at Seoul National University Hospital.[br]Patients. The study group included 45 males and 41 females with a mean age of 50.67 12.62 years.[br]Interventions. All patients underwent adrenal computed tomography (CT) and AVS.[br]Main Outcome Measures. Main outcomes measures included the accurate localization of lesions accountable for PA.[br]Results. The success rate of AVS was 83.7% (87/104) for right adrenal vein, 94.2% (98/104) for left adrenal vein and 82.69% (86/104) for overall. AVS revealed bilateral aldosterone hypersecretion in 15/75 patients with unilateral abnormalities diagnosed by CT. These patients underwent medical treatment instead of surgery. One patient had an adrenal mass on right side but AVS lateralized aldosterone secretion to the left side. This patient underwent left adrenalectomy. Furthermore, AVS revealed unilateral lesions in 2/5 patients with bilateral lesions demonstrated by CT. These patients underwent unilateral adrenalectomy. Finally, AVS demonstrated lateralization in 1/6 of patients with no CT abnormalities, who were subjected to surgery.[br]Fifty-three patients with unilateral hypersecretion and one patient with bilateral hypersecretion underwent surgical removal of the affected gland(s). All patients had resolution of hypokalemia and clinical improvement of hypertension.[br]Conclusions. Many patients (19/86, or 22.09%) would have been inappropriately managed if decision-making were made solely on CT findings. Therefore, AVS is recommended before determining definitive PA management.[br][br]Nothing to Disclose: HJC, YJK, EMO, JHK, KEL, SYK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 601 194 1514 SUN-489 PO01-01 Sunday 1311 2012


1308 ENDO12L_SUN-490 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Association between Primary Hyperaldosteronism and Pheochromocytoma: A Diagnostic and Therapeutic Challenge Rosa Maria Paragliola, Maria Pia Ricciato, Carmela De Crea, Vittoria Rufini, Guido Fadda, Alfredo Pontecorvi, Rocco Bellantone, Salvatore M Corsello Catholic University School of Medicine, Rome, Italy; Catholic University School of Medicine, Rome, Italy; Catholic University School of Medicine, Rome, Italy; Catholic University School of Medicine, Rome, Italy Secondary hypertension represents only about 5-10% of all hypertensions in adults. A proper diagnosis is essential because underlying causes are potentially correctable, especially pheochromocytoma (Pheo) and primary hyperaldosteronism (PA). The association between these two conditions is very rare. A 55 year-old man came to our attention in march 2010 for hypertension with hypokaliemia (serum potassium 2.8 mEq/L). He had hypertensive crisis (220/120 mmHg) with headache, swelling and palpitation. Biochemical evaluation showed low plasmatic renin levels (1.2 pg/ml) with high plasmatic aldosterone (460 ng/ml), suggestive of PA. Abdomen CT scan showed a bilateral adrenal hyperplasia with a 18 mm right adenoma. Adrenal venous sampling confirmed a PA due to a bilateral adrenal hyperplasia, because there was no ACTH-stimulated aldosterone/cortisol gradient between right and left gland. 24-hrs urine collection showed high levels of urinary norepinephrines (265 mcg/day, n.v. 12-86) and normetanephrines (2704 mcg/day, n.v. 90-445) with normal epinephrines (7.3 mcg/day, n.v. 2-22) and metanephrines (200 mcg/day, n.v. 50-340). [sup]123[/sup]I-MIBG scintigraphy, performed in the suspicion of Pheo, was negative. Patient started therapy with spironolactone, doxazosine, metoprolol and potassium chloride with periodical evaluation and posological adjustments. Blood pressure was poorly controlled by medical therapy and urinary norepinephrines and normetanephrine resulted constantly high in spite of a second negative [sup]123[/sup]I-MIBG scintigraphy. In October 2011 a [sup]18[/sup]F-FDOPA PET was performed, showing a clear right adrenal monolateral uptake suggestive of Pheo. Thus, in January 2012, the patient underwent right adrenalectomy. Histology showed [ldquo]pheochromocytoma (pheochromocytes positive for chromogranin A and synaptophysin, negative for CAM 5.2 with cellular elements positive for S-100. Ki 67 [lt] 1%). Surrounding adrenal gland slightly enlarged, in particular in glomerulosa, with several micronodules[rdquo]. Therefore final diagnosis confirmed [ldquo]pheochromocytoma with micronodular cortical hyperplasia[rdquo]. This case represents an association between two rare forms of endocrine hypertensions. The best treatment of Pheo, if confirmed by imaging, is surgery while PA, especially if due to a bilateral adrenal hyperplasia, can be treated with spironolactone. In our case [sup]18[/sup]F-FDOPA PET is confirmed to be the gold standard in the diagnosis of Pheo, being able to detect [sup]123[/sup]I-MIBG negative tumors.[br][br]Nothing to Disclose: RMP, MPR, CDC, VR, GF, AP, RB, SMC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1121 194 1515 SUN-490 PO01-01 Sunday 1312 2012


1309 ENDO12L_SUN-491 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Referral Pattern [amp] Biochemical Work-Up of Adrenal Lesions and the Role of Endocrinologists in Managing Patients Undergoing Surgical Adrenalectomy: A Single Center Audit of 10 Years of Laparoscopic Adrenalectomies Ketan Dhatariya, Azin Aghili, Sudhanshu Chitale, Jim Armitage, Frankie Swords, Neil Burgess Norfolk and Norwich University Hospitals NHS Foundation Trust, Norwich, UK; University of East Anglia, Norwich, UK; Norfolk and Norwich University Hospitals NHS Foundation Trust, Norwich, UK [bold]Introduction:[/bold] Laparoscopic adrenalectomy is the gold standard treatment for adrenal lesions. High volume centres with input from endocrinologists and urologists with a special interest in adrenal lesions may have best outcomes.[br][bold]Aims: [/bold]To audit our practice and set standards with reference to preoperative workup and postoperative follow-up. This is a single centre, single surgeon experience of surgical adrenalectomy.[br][bold]Material [amp] Methods: [/bold]Over a 10 year period 71 adrenalectomies were performed. 55 clinical notes were available for review. M:F 24:31, with equal gender distribution amongst all age groups except the 21-30 yr group (M: F 0:4). Most patients (n=18) were in the 6[sup]th[/sup] decade. 33/55 cases were referred by endocrinologists, 13 from other medical specialities, 8 by other surgical specialties, with a single direct primary care referral. 24 patients had left sided, 24 had right sided solitary lesions, and 7 cases underwent bilateral adrenalectomy.[br][bold]Results: [/bold]The formal adrenal CT protocol was followed in only 10/55 cases. Input from an endocrinologist was available in 44/55 cases preoperatively and 33/55 postoperatively. 11/55 were discussed in the relevant specialty MDT. Whilst most patients have some form of endocrine assessment, few had all of the appropriate tests done.[br]The urologist followed up 44/55 cases within 16 weeks. The endocrinologists had no follow up for 17 cases.[br][bold]Conclusion: [/bold]In the previous 10 years, patients presenting to our service with adrenal lesions do not appear to have received the service we would hope them to have. Adrenal protocol CTs are frequently not performed for incidental lesions and full endocrine assessment is not always performed preoperatively. We have set standards to tighten our processes since this audit was carried out.[br][br]Nothing to Disclose: KD, AA, SC, JA, FS, NB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 12 194 1516 SUN-491 PO01-01 Sunday 1313 2012


1310 ENDO12L_SUN-492 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Allelic Frequencies of [italic]HLA-A[/italic], [italic]HLA-B[/italic] and [italic]HLA-DRB1[/italic] Genes in Patients with Adrenocortical Tumor Carriers of the Germline Mutation R337H in the [italic]TP53 [/italic]Gene Alexandre Menna Barreto Cordeiro, Maria da Graca Bicalho, Rosana Marques-Pereira, Patricia Pinho Franca, Marcio Marques Moraes, Gorete Ynaquievi Tomaz Rezende, Suzana Nesi-Franca, Romolo Sandrini, Luiz De Lacerda Federal University of Parana, Curitiba, Brazil; Federal University of Parana, Curitiba, Brazil [bold]Context[/bold]. Nearly 90% of the children and adolescents with adrenocortical tumors (ACT) in Paran[aacute], Brazil, are carriers of the germline mutation R337H in the [italic]TP53[/italic] gene. However, the mutation alone is not sufficient for the development of the tumor. Close association of certain types of malignancies, such as osteosarcoma, leukemia and Hodgkin[apos]s disease, and the HLA system has been documented and recent studies have shown reduced expression of class II MHC complex genes in adrenocortical carcinomas.[br][bold]Objectives[/bold]. Evaluate the allelic frequency of the [italic]HLA-A[/italic],[italic] HLA-B [/italic]e [italic]HLA-DRB1[/italic] genes in ACT patients with the R337H germline mutation of the [italic]TP53 [/italic]gene, and compare these results with relatives carriers of the same mutation and controls.[br][bold]Design[/bold]. Genomic study.[br][bold]Patients and Participants.[/bold] 39 ACT patients followed up regularly at the Pediatric Endocrine Unit of the Federal University of Paran[aacute] School Hospital, 47 relatives of the probands and 64,219 controls.[br][bold]Intervention[/bold]. [italic]HLA [/italic]typing.[br][bold]Main Outcome Measure[/bold]. Frequency of allelic variants of the [italic]HLA-A[/italic],[italic] HLA-B [/italic]e [italic]HLA-DRB1[/italic] genes.[br][bold]Results. [/bold]The allelic variants [italic]HLA-A[/italic]*[italic]31[/italic], [italic]HLA-B[/italic]*[italic]14[/italic], [italic]HLA-B[/italic]*[italic]39[/italic] e [italic]HLA-DRB1[/italic]*[italic]08[/italic] were significantly more frequent in patients compared with controls. Subgroup analysis showed that the allelic groups [italic]HLA-A[/italic]*[italic]68[/italic], [italic]HLA-B[/italic]*[italic]39[/italic] e [italic]HLA-DRB1[/italic]*[italic]08[/italic] were more frequent in patients with stages I and II than in controls, whereas the [italic]HLA-A[/italic]*[italic]31[/italic], [italic]HLA-A*68[/italic] e [italic]HLA-B[/italic]*[italic]39[/italic] allelic variants were significantly more frequent in patients with carcinoma than in controls.[br][bold]Conclusion. [/bold]These results suggest an association between the HLA system and the development of ACT in children and adolescents with the [italic]TP53[/italic] gene germline mutation R337H.[br][br]Nothing to Disclose: AMC, MdGB, RM-P, PPF, MMM, GYTR, SN-F, RS, LDL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 461 194 1517 SUN-492 PO01-01 Sunday 1314 2012


1311 ENDO12L_SUN-493 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) The Wnt/[beta]-Catenin and Ras/Raf/MEK/ERK Signaling Pathways Alterations in Adrenocortical Tumors Beatrice Rubin, Raffaele Pezzani, Susi Barollo, Barbara Mariniello, Maria Verena Cicala, Monica Salva, Maurizio Iacobone, Ambrogio Fassina, Franco Mantero University of Padua, Padua, Italy; University of Padua, Padua, Italy; University of Padua, Padua, Italy Adrenocortical tumors (ACT) include benign and malignant tumors. Adrenocortical carcinomas (ACC) are highly malignant neoplasms with a poor prognosis, but their genetic alterations to date identified are limited. Laboratory studies on ACT have revealed a wide variety of signaling pathways involved in these tumors, among these Wnt/[beta]-catenin signaling pathway and Ras/Raf/MEK/ERK pathway resulted often disregulated. Another important factor in many signaling pathways is the epidermal growth factor receptor (EGFR), responsible for proliferation and overexpressed in many adrenocortical tumors.[br]The objective of our study was to evaluate genetic alterations in key components of Wnt/[beta]-catenin and Ras/Raf/MEK/ERK signaling pathways in order to better understand the pathogenesis of sporadic adrenocortical tumors and provide light onto new possible prognostic factors.[br]We performed high resolution melting (HRM) analysis for evaluating the presence of activating mutations in[italic] EGFR[/italic] (exons 18, 19, 20, 21), [italic]BRAF[/italic] (exons 11 and 15), [italic]H-RAS[/italic] (exons 1 and 2), [italic]N-RAS[/italic] (exons 1 and 2), [italic]K-RAS[/italic] (exons 1 and 2), [italic]CTNNB1[/italic] (exon 3), [italic]AXIN2 [/italic](exon 7). We analyzed a series of 92 sporadic samples: 21 ACC, 38 aldosterone producing adenomas (APA), 29 cortisol producing adenomas (CPA) and 4 normal adrenocortical tissues. Only samples resulted with altered melting curves were direct sequenced.[br]We found 2 different [italic]BRAF[/italic] mutations in 2 ACC, 4 [italic]H-RAS[/italic] silent mutations in 1 APA, 1 CPA and in 2 ACC, 16 [italic]CTNNB1[/italic] alterations in 5 APA, 6 CPA and 5 ACC. No alteration in [italic]EGFR[/italic], [italic]N-RAS[/italic], [italic]K-RAS[/italic].[br]These results suggest that abnormalities in Ras/Raf/MEK/ERK pathway do not represent a frequent pathogenetic mechanism of adrenocortical tumorigenesis, while alterations in Wnt/[beta]-catenin pathway seem to be a more common event. Nevertheless this study identified diverse genetic alterations, whose role in adrenocortical tumors should be further investigated.[br][br]Sources of Research Support: ENSAT Cancer. Grant number 259735.[br][br]Nothing to Disclose: BR, RP, SB, BM, MVC, MS, MI, AF, FM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 333 194 1518 SUN-493 PO01-01 Sunday 1315 2012


1312 ENDO12L_SUN-494 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Integrative Analysis of Genome-Wide Epigenetic and Gene Expression Analysis Reveals Differential Signaling Pathway Expression in Adrenocortical Cancer Maria Gueorguiev, Christina Thirlwell, Phil East, Candy Sze, Dan Berney, Robert Carpenter, Scott Akker, William Drake, Shern Chew, Stephan Beck, Ashley Grossman St Bartholomew[apos]s Hospital, Queen Mary University of London, London, UK; William Harvey Research Institute, Queen Mary University of London, London, UK; Royal Free Hospital, London, UK; University College London, London, UK; Cancer Research UK, London, UK; Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, UK; St Bartholomew[apos]s Hospital, Queen Mary University of London, London, UK; Churchill Hospital, University of Oxford, Oxford, UK Introduction: Dysregulation of signaling pathways orchestrates the growth and survival/progression of many cancers, and may relate to specific DNA methylation and gene expression levels. Our aim was to perform genome-wide DNA methylation and mRNA gene expression array study and integrate the data in order to establish biomarkers for the early detection and identification of new therapeutic targets for adrenocortical cancer (ACC). This preliminary study reports on epigenetic and gene expression profiling that has identified potential new pathways involved in ACC tumorigenesis.[br]Patients and Methods: Ten fresh-frozen specimens including 7 sporadic adrenocortical tumors (2 malignant, 5 benign) and 3 adjacent normal adrenal cortices were analysed with both Illumina HumMeth27 beadarray (which interrogates 27,500 CpG sites of 14,000 genes) and the Affymetrix HumanGene 1,0 ST mRNA expression array (which uses a mean of 26 probes covering the coding region of 28,869 genes). The DNA methylation and gene expression pattern was correlated with the tumor phenotype using logistic regression analysis.[br]Results: Integrated genome-wide analysis revealed differential DNA methylation status and mRNA expression levels of highly malignant versus low grade malignant or versus benign adrenocortical tumours. DNA methylation features and gene expression showed also a distinct pattern between malignant adrenocortical tumors and normal tissue samples. The methylation status correlated with mRNA expression. Dysregulated pathways include the IGF-2 pathway, which has been shown to be overexpressed in ACC. Dysregulation of components involved in the cell cycle, mitosis and spindle organisation and cytokinesis, regulation of centrosomes as well as in chromosome instability (CDKN3, CCNE1, ASPM, BUB1, KIF11, MAD2L1, TOP2A, SEPT4, KIAA0101) but also S100B, was also observed. The fold expression variation across the genes extends from -2.92 to +4.57.[br]Conclusion: We have applied a novel approach to integrate epigenetic and gene expression data for adrenocortical tumors. This integrated approach is a strong tool that has demonstrated the involvement of the cell cycle/spindle organisation and cytokinesis network as a differential pathway for ACC pathogenesis and progression, and may represent a target for future therapy. DNA methylation is a regulatory mechanism responsible for the differing mRNA expression among various adrenocortical tumors.[br][br]Nothing to Disclose: MG, CT, PE, CS, DB, RC, SA, WD, SC, SB, AG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1935 194 1519 SUN-494 PO01-01 Sunday 1316 2012


1313 ENDO12L_SUN-495 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) New Insights into the Role of LIN28A and LIN28B in Adrenocortical Tumorigenesis of Pediatric and Adult Patients Andre M Faria, Beatriz MP Mariani, Ibere C Soares, Tamaya C Ribeiro, Antonio M Lerario, Daniel S Freire, Alda Wakamatsu, Rodrigo A Ressio, Venancio AF Alves, Maria C Zerbini, Berenice Bilharinho Mendonca, Maria CBV Fragoso, Ana C Latronico, Madson Q Almeida Hospital das Cl[iacute]nicas, Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Hospital das Cl[iacute]nicas, Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil LIN28, a stem cell-expressed RNA-binding protein, has emerged as an important modulator of cell reprogramming and pluripotency through both microRNA-dependent and independent pathways. Overexpression of LIN28 has been demonstrated in many different human tumors with an overall frequency of 15%. We have therefore examined the expression of LIN28 in pediatric and adult adrenocortical tumors (ACT). LIN28A and LIN28B expression were assessed in 87 ACT (25 children and 62 adults) and in a commercial pool of normal adrenal cortex by real time PCR. LIN28A expression was 8 times more expressed than LIN28B in the pool of normal adrenal cortex at mRNA level. LIN28A overexpression at mRNA level was more frequent in ACT with Weiss score [ge] 3 (64%) than in those tumors with Weiss [lt] 3 (19%; X2= 11.8, p=0.001) from adult patients, but was not correlated with overall and recurrence-free survival (p= 0.19 and p= 0.74, respectively). In children, LIN28A overexpression was not significantly different according to Weiss score. LIN28B was mainly expressed at mRNA level in pediatric ACT (68%, 17/25) when compared to adult ACT (6%, 4/62; X2= 36.9, p[lt] 0.0001). In addition, LIN28 staining was evaluated in a tissue microarray including 104 ACT (37 children and 67 adults). LIN28 protein expression was significantly higher in non-metastatic ACT with Weiss [ge] 3 in comparison with metastatic ACT and ACT with Weiss [lt] 3 (p[lt] 0.05). Interestingly, the lower expression of LIN28 observed in metastatic ACT (80%, 16/20) when compared to non-metastatic ACT (43%, 36/84; X2= 8.9, p= 0.003) may suggest a defective translational control of protein expression in the former group. Among ACT with Weiss [ge] 3, a weak LIN28 expression was correlated with reduced overall survival at the univariate analysis (p= 0.04, hazard ratio (HR) 0.26, confidence interval (CI) 0.07-0.96) and was an independent predictor of recurrence-free survival in ACT with Weiss [ge] 3 (p= 0.001, HR 0.05, CI 0.005-0.55) at multivariate analysis. LIN28A and LIN28B deletion or amplification was not identified by multiplex ligation-dependent probe amplification in 14 ACT. In conclusion, low expression of LIN28 at protein level was a hallmark of metastatic ACT. Additionally, LIN28B, a marker of germ cell tumors, was almost exclusively expressed in pediatric ACT. Therefore, our findings bring new insights into the role of LIN28A and LIN28B in pediatric and adult adrenocortical tumorigenesis.[br][br]Sources of Research Support: FAPESP (2011/09092-0).[br][br]Nothing to Disclose: AMF, BMPM, ICS, TCR, AML, DSF, AW, RAR, VAFA, MCZ, BBM, MCBVF, ACL, MQA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1579 194 1520 SUN-495 PO01-01 Sunday 1317 2012


1314 ENDO12L_SUN-496 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Silencing of IGF1R by Small Interfering RNA (siRNA) in an Adrenocortical Tumor Cell Line Tamaya Castro Ribeiro, Luciana Ribeiro Montenegro, Alexander Augusto Jorge, Miriam Yumie Nishi, Berenice Bilharinho Mendonca, Ana Claudia Latronico Hospital das Clinicas da Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Background: Sporadic adrenocortical tumors are frequently diagnosed as incidentalomas in adult individuals, but these tumors are rare in children. However, a remarkably high prevalence of adrenocortical tumors has been reported in children from Southern Brazil. Overexpression of IGF2 and/or IGF1R may trigger a cascade of molecular events that can ultimately lead to adrenocortical malignancy. The inhibition of IGF1R expression can be a useful tool in the treatment of adrenocortical tumors.[br]Objective: To induce IGF1R gene silencing by siRNA in a human adrenocortical tumor cell line (NCI H295).[br]Methods: The human adrenocortical tumor NCI H295 line was cultured in RPMI 1640 medium. All experiments were carried out in four groups: 1) untreated NCI H295 cells, 2) NCI H295 cells transfected with a negative control, 3) NCI H295 cells transfected with specific IGF1R siRNA # 1 (exon 18) and 4) NCI H295 cells transfected with specific IGF1R siRNA # 2 (exon 2). IGF-1R gene and protein expression were determined by the techniques of real-time PCR and Western blot, respectively.[br]Results: The two specific siRNA for IGF1R had similar efficiency and were able to reduce IGF-1R expression both at the messenger RNA and protein levels. As expected, the untreated cells and the negative control groups exhibited equivalent mRNA and protein expressions. The expression inhibition rate of IGF1R mRNA levels was 50% at 48 hours after transfection. Western blot detected a 30% decrease in IGF-1R protein levels in cells transfected with both siRNAs # 1 and # 2.[br]Conclusion: Specific siRNAs inhibited IGF1R expression both at mRNA and protein levels. The silencing of IGF-1R expression represents a potential target and has a great interest in obtaining new and effective therapeutic options for human adrenocortical tumors.[br][br]Nothing to Disclose: TCR, LRM, AAJ, MYN, BBM, ACL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1466 194 1521 SUN-496 PO01-01 Sunday 1318 2012


1315 ENDO12L_SUN-497 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Effects of 1,25-Dihydroxyvitamin D on the Human Adrenocortical Carcinoma Cell Line NCI-H295R Catia Pilon, Riccardo Urbanet, Silvia Vettore, Rossella Sirianni, Vincenzo Pezzi, Francesco Fallo University of Padova, Padova, Italy; University of Calabria, Arcavacata di Rende-Cosenza, Italy Vitamin D receptor (VDR) and its ligand 1,25-Dihydroxyvitamin D3 (1,25OHD3) play, in general, an inhibitory role the growth of normal and malignant cells. However, the mechanisms for this anti-proliferative action remain not completely understood. Recently, a 1,25OHD3-mediated effect on hormone production and steroidogenic genes has been reported in the human steroid-secreting adrenocortical cancer cell line NCI-H295R (Lundqvist J et al., 2010). The aim of our study was twofold: 1) to test expression of vitamin D metabolism genes in normal adrenals and in a series of adrenal tumors as well as in the NCI-H295R cells; 2) to investigate the potential anti-proliferative action of 1,25OHD3 on NCI-H295R cells underlying the molecular mechanisms behind this effect. mRNA levels for CYP2R1 (i.e. the enzyme converting vitamin D3 to 25OHD3), CYP27B1 (i.e. the enzyme converting 25OHD3 to 1,25OHD3) and VDR were measured by RT-PCR and/or Western blotting in both NCI-H29R cells and adrenal tissues. DNA synthesis was evaluated according to [3H]TdR cell incorporation after 96 hours treatment of NCI-H295R cells with 1,25OHD3 at increasing doses, in comparison to untreated control cells. The effect of 1,25OHD3 on cell apoptosis and cell cycle was analyzed with a flow cytometer. CYP2R1, CYP27B1 and VDR were expressed in NCI-H295R cells and in all adrenal tissues analyzed (non-functioning adenomas, n=5; cortisol/aldosterone-secreting adenomas, n=9; cortisol-producing carcinoma, n=1. 1,25OHD3 inhibited cell proliferation by 20% at a dose of 10[sup]-8[/sup]M and induced a concomitant decrease in aldosterone and DHEA-S production. 1,25OHD3 induced cell cycle arrest, promoting accumulation of cells in G0/G1 phase without inducing apoptosis. Conclusions: 1,25OHD3 has cytotoxic effects on the NCI-H295 cells by promoting cell cycle arrest. Expression of vitamin D metabolism genes in NCI-H295R cells as well as in normal and tumor adrenals suggests a potential paracrine role of vitamin D in adrenal growth and function. Because of its anti-proliferative action, 1,25OHD3 could be considered for the treatment of patients with adrenocortical cancer.[br][br]Nothing to Disclose: CP, RU, SV, RS, VP, FF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 128 194 1522 SUN-497 PO01-01 Sunday 1319 2012


1316 ENDO12L_SUN-498 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Determination of the Oncogenic Ras Signaling Requirement for Adrenocortical Tumorigenesis Aude Salomon, Flavie Sicard, Maryline Herbet, Jean-Jacques Feige, Michael Thomas iRTSV/BCI, CEA, Universit[eacute] J Fourier, Grenoble, France Much of the significant advances in identifying genes involved in sporadic adrenocortical carcinoma (ACC) development derive from the study of human tumor specimens. To complement these studies, several laboratories including ours have developed murine models of ACC. Such models provide an important basis for understanding the specific genetic alterations that lead to particular phenotypes. Thus, we have shown that the successive infection of normal primary bovine adrenocortical cells with the oncogenic allele RasV12 and then by a truncated form of p53 (p53DD) resulted in cells that formed tumors with a metastatic behavior. In contrast, the transplantation of cells infected with either construct alone produced a normal tissue in the case of the p53DD and an hyperplasia in the case of RasV12.[br]Hence, we asked which of the Ras effector pathways (Raf, RalGEF or PI3K) is required to initiate adrenocortical tumor growth. We generated three cell populations expressing RasV12S35, RasV12G37 or RasV12C40 in which the above-mentioned pathways were respectively and primarily activated. Following transplantation into SCID mice, RasV12S35 cells formed a large benign tissue with no signs of kidney invasion. The tissue was highly cellular with a uniform structure of regular eosinophilic cells. The Ki-67 expression in tissues showed a labelling index of 9.27 [plusmn] 0.82% which represents half of the index measured in tissues expressing RasV12 (18.72 [plusmn] 3.25%). The RasV12G37 cells formed a smaller tissue with an irregular architecture, cellular pleiomorphism and nuclear atypia. The proliferation index was much lower (3.5 [plusmn] 1.2%) and was similar to that in tissues formed following transplantation of control cells (3.51 [plusmn] 1.04%). As concerns RasV12C40 cells, they formed small tissues with necrotic areas with almost no proliferating cells (0.25 [plusmn] 0.12%). We then infected each cell population with p53DD and studied their capacity to form tumors once transplanted. We found that the Raf, but not the RalGEF or PI3K pathway, was sufficient for Ras transformation of adrenocortical cells. We noticed that after 9 weeks in vivo the RasV12S35/p53DD tumors invaded the kidney but no metastasis was detectable unlike the RasV12/p53DD tumors.[br]Thus, oncogenic Ras appears to transform adrenocortical cells mainly through the activation of the Raf pathway. However, the full metastatic phenotype requires probably a synergistic cooperation between the Raf and either RalGEF or PI3K pathway or, both.[br][br]Sources of Research Support: Association pour la Recherche sur le Cancer, Ligue Contre le Cancer (Comit[eacute]s de la Loire et de l[apos]Is[egrave]re).[br][br]Nothing to Disclose: AS, FS, MH, J-JF, MT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 491 194 1523 SUN-498 PO01-01 Sunday 1320 2012


1317 ENDO12L_SUN-499 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Adhesion to Type IV and V Collagens Sensitizes Staurosporine-Induced Apoptosis in H295R Adrenocortical Cancer Cells Roberto Vicinanza, Tiziana Nardo, Geraldina Micalizzi, Antonio Stigliano, Vincenzo Toscano, Susanna Scarpa Sapienza University of Rome, Rome, Italy; Sapienza University of Rome, Rome, Italy Adrenocortical carcinoma (ACC) is a rare and aggressive tumor characterized by poor prognosis and resistance to conventional chemotherapeutic agents. Tumor progression results, in part, from the dynamic interaction between cancer cells and extracellular matrix (ECM) and this can modulate cell proliferation and apoptosis. In this study we investigated whether adhesion to different ECM components affected staurosporine (ST)-induced apoptosis in H295R adrenocortical cancer cells. H295R cells were exposed for 12 h to ST (1[mu]M) in the absence or presence of different substrates. The adhesion of H295R cells to type IV (C-IV) and type V (C-V) collagens significantly increased the antiproliferative effect of ST compared to ST alone (p[lt]0.01), and these results were associated with morphological cell modification. In contrast, the adhesion to polilisine, matrigel, laminin, fibronectin, type I and type III collagens did not enhance the ST effect. We further demonstrated that pretreatment with anti-integrin alpha 2 antibody suppressed C-IV and C-V effects, suggesting that adhesion of H295R cells to these specific substrates may modulate the antiproliferative activity of ST. Western blot analysis of cleaved poly (ADP-ribose) polymerase (PARP), and immunofluorescence staining for Annexin V/propidium iodide and active caspase-3 showed that ST-induced apopotosis was significantly increased in cells adhering to C-IV and C-V compared to ST alone (p[lt]0.01) and pretreatment with anti-integrin alpha 2 antibody abrogated this effect. These data demonstrate that adhesion of H295R cells to type IV and V collagens sensitizes ST-induced apoptosis and provide a further understanding of adrenocortical carcinoma by delineating the role of ECM components, not only in supporting tissue architecture, but also in modulating cell proliferation and perhaps the response to chemotherapeutic agents.[br][br]Nothing to Disclose: RV, TN, GM, AS, VT, SS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1845 194 1524 SUN-499 PO01-01 Sunday 1321 2012


1318 ENDO12L_SUN-500 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Ouabain Reduced Proliferation in Adrenocortical Cell Lines and in Primary Adrenocortical Tumor Cells Raffaele Pezzani, Beatrice Rubin, Susi Barollo, Marco Redaelli, Carla Mucignat-Caretta, Franco Mantero University of Padova, Padova, Italy; University of Padova, Padova, Italy Ouabain is a cardiotonic steroid obtained from Strophanthus species. Recently its role as antiproliferative agent has been investigated in tumor cells. Adrenocortical carcinoma (ACC) is a rare cancer, with poor prognosis, of which SW13 and H295R cell lines could be considered a cellular model. Our aim was to evaluate if ouabain exerts anticancer properties.[br]The effects of ouabain were assessed by MTT assay, by [3H] thymidine assay, by Wright[apos]s staining method, by flow cytometry analysis, by homogeneous caspase assay and by Western blot. Ouabain induced a reduction in cell viability at 24h and 72h in SW13, in H295R and 4 primary adrenocortical tumor cell cultures. Proliferation rate decreased effectively in SW13 and H295R at higher doses. Marked morphological changes were observed both for SW13 and H295R cell lines, with an increase in necrotic process. Cell cycle distribution was altered by ouabain in SW13. Apoptosis analysis demonstrated an increase in caspase activity, clearly evident for SW13 at 72h. FACS analysis by Annexin V-FITC and propidium iodide confirmed an increase in necrotic process especially at higher concentrations. Western blot analysis showed that Erk, Akt, p-Akt, P70s6k, p-P70s6k and VEGF decreased after ouabain treatments in SW13.[br]In conclusion ouabain exerts antiproliferative effects on SW13 and H295R cell lines and on primary adrenocortical tumor cells. Therefore, these data support the evidence of ouabain as a potential anticancer agent.[br][br]Sources of Research Support: The AIRC association (Associazione Italiana Ricerca sul Cancro) grant number 5250;. supported by ENS@T-CANCER (European Network for the Study of Adrenal Tumours) grant number 259735.[br][br]Nothing to Disclose: RP, BR, SB, MR, CM-C, FM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 379 194 1525 SUN-500 PO01-01 Sunday 1322 2012


1319 ENDO12L_SUN-501 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Activation of GPER Inhibits Growth of Adrenocortical Cancer (ACC) Cells [italic]In Vitro[/italic] and [italic]In Vivo[/italic], Suggesting a New Therapy for ACC Rosa Sirianni, Adele Chimento, Arianna De Luca, Carmen Ruggiero, Francesco Fallo, Catia Pilon, Giorgio Arnaldi, Giulia Carpinelli, Lidia Cerquetti, Antonio Stigliano, Vincenzo Pezzi University of Calabria, Rende, Italy; University of Padua, Padua, Italy; University of Ancona, Ancona, Italy; Istituto Superiore di Sanit[agrave], Rome, Italy; University of Rome [quot]Sapienza[quot], Rome, Italy Our previous study indicated a central role for Estrogen receptor (ER) alpha (ESR1) in both estrogen-dependent and -independent (induced by IGF-II/IGF1R pathways) proliferation of ACC cells. In fact, treatment with Selective Modulator of Estrogen Receptor Tamoxifene (Tam) significantly suppressed the growth of adrenocortical carcinoma H295R xenograft model. In this study we investigated the role of G-protein-coupled estrogen receptor (GPER), a membrane ER, on ACC cell proliferation. We revealed GPER mRNA and protein expression in human ACC tissues as well as in H295R cells. Surprisingly, we found that activation of GPR30 by the receptor-specific, non-estrogenic ligand, G-1 inhibited the growth of adrenocortical tumor H295R cells in vitro and H295R xenografts in vivo. Moreover, treatment of H295R cells with G-1 induced DNA damage and apoptosis. Silencing GPER with a specific siRNA significantly blocked G-1 effects on H295R cell proliferation and apoptosis confirming a specific involvement of GPER in pathway(s) inhibiting ACC growth. Experiments focused on clarifying the molecular mechanism underlying G-1-induced inhibition of ACC growth are currently on going. Preliminary results suggest the involvement of sustained activation of ERK1/2 and calcium signaling. In conclusion these results, together with our previous studies, suggest that combined treatments with both SERM and GPER agonists could be an effective new therapy for controlling ACC growth.[br][br]Sources of Research Support: Associazione Italiana Ricerca sul Cancro IG10344.[br][br]Nothing to Disclose: RS, AC, ADL, CR, FF, CP, GA, GC, LC, AS, VP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1247 194 1526 SUN-501 PO01-01 Sunday 1323 2012


1320 ENDO12L_SUN-502 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Abnormal Serotonergic Regulatory Loop in Primary Pigmented Nodular Adrenal Disease (PPNAD) Tissues Associated with Cushing Syndrome: Role of the Activation of the PKA Pathway Zakariae Bram, Sylvie Renouf, Celine Duparc, Bruno Ragazzon, Rossella Libe, Antoine Martinez, Constantine Stratakis, Jerome Bertherat, Estelle Louiset, Herve Lefebvre University of Rouen, Mont Saint Aignan, Rouen, France; Cochin University Hospital, APHP, Paris, France; GRE, University of Clermont, Clermont Ferrand, France; National Institutes of Health, National Institute of Child Health and Human Development, Bethesda, MD; University Hospital of Rouen, Rouen, France In the normal adrenal gland, serotonin (5-HT) stimulates cortisol secretion through activation of 5-HT4 receptors whereas, in some macronodular adrenal hyperplasia tissues, the corticotropic effect of 5-HT is mediated by ectopic 5-HT7 receptors. The aim of the present study was to investigate the role of 5-HT in the control of cortisol secretion in PPNAD tissues from 12 patients by using molecular, immunohistochemical and pharmacological approaches. RT-PCR studies revealed overexpression of the genes encoding tryptophan hydroxylase, the key enzyme of the synthesis of 5-HT, and the 5-HT4 and 5-HT7 receptors in comparison with normal adrenals. Tryptophan hydroxylase was detected in hyperplastic nodules by immunohistochemistry. 5-HT dose-dependently increased cortisol production by PPNAD cells derived from 6 patients. The potency and efficacy of 5-HT to stimulate cortisol were higher in PPNAD than in normal adrenocortical cells. 5-HT was also able to increase expression of the CYP11B1 gene, which encodes the steroidogenic enzyme 11alpha-hydroxylase, in the PPNAD immortalized cell line LT2. Both the 5-HT4 receptor antagonist GR113808 and the 5-HT7 receptor antagonist SB269970 significantly inhibited the stimulatory effect of 5-HT on cortisol by both reducing the potency and efficacy of the indolamine. These data indicate that, in PPNAD cells, the stimulatory action of 5-HT on cortisol release is mediated by both the eutopic 5-HT4 receptor and an ectopic 5-HT7 receptor. In most patients, PPNAD results from inactivating mutations of the PRKAR1A gene which cause enhancement of the PKA pathway. In the adrenocortical cell line H295R, inhibition of PRKAR1A gene expression markedly stimulated expression of tryptophan hydroxylase, 5-HT4 and 5-HT7 receptor mRNAs. Taken together, our results show that activation of the PKA pathway triggers formation of an illicit serotonergic regulatory loop in PPNAD tissues associated with Cushing[apos]s syndrome.[br][br]Sources of Research Support: This work was supported by INSERM U982, the Carney Complex network and ANR Genopat 2008.[br][br]Nothing to Disclose: ZB, SR, CD, BR, RL, AM, CS, JB, EL, HL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 750 194 1527 SUN-502 PO01-01 Sunday 1324 2012


1321 ENDO12L_SUN-503 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Ectopic Intraadrenal Production of ACTH in Macronodular Bilateral Adrenal Hyperplasia Causing Cushing Syndrome: Role in the Control of Cortisol Secretion Estelle Louiset, Celine Duparc, Jacques Young, Isabelle Boutelet, Sylvie Renouf, Zakariae Bram, Lionel Groussin, Philippe Caron, Antoine Tabarin, Fabienne Grunenberger, Sophie Christin-Maitre, Jean-Marc Kuhn, Youssef Anouar, Jerome Bertherat, Herve Lefebvre Institute for Research and Innovation in Biomedicine (IRIB), Rouen University, Mont-Saint-Aignan, France; Institut Gustave Roussy, Le Kremlin-Bic[ecirc]tre, France; Cochin University Hospital, APHP, Paris, France; Centre Hospitalier Universitaire Larrey, Toulouse, France; H[ocirc]pital Haut L[eacute]v[ecirc]que, Centre Hospitalier Universitaire de Bordeaux, Pessac, France; H[ocirc]pital de Hautepierre, Strasbourg, France; H[ocirc]pital Saint-Antoine, Assistance Publique H[ocirc]pitaux de Paris, Paris, France; Rouen University Hospital, Rouen, France Illicit expression of membrane receptors for circulating regulatory factors, such as gastric inhibitory polypeptide (GIP), luteinizing hormone (LH) and serotonin (5-HT) receptors, has been well documented in ACTH-independent macronodular adrenal hyperplasias (AIMAHs) causing Cushing[apos]s syndrome. In addition, we have observed an abnormal expression of ACTH in some steroidogenic cells in two AIMAH tissues. The aim of the present study was to investigate the role of local production of ACTH in the control of steroidogenesis in a series of 30 AIMAH tissues. Expression of pro-opiomelanocortin (POMC) mRNA and ACTH-like immunostaining were detected in all tissues studied. ACTH co-localized with 17-hydroxylase, the HDL-cholesterol receptor SR-B1, prohormone convertase 1 and secretogranin II immunoreactivities in clusters of cells disseminated throughout hyperplasia tissues. Perifusion experiments demonstrated that adrenal slices spontaneously released detectable amounts of ACTH in a pulsatile fashion. ACTH secretion was significantly increased in vitro by GIP, hCG and 5-HT in tissues previously sensitive in vivo to the stimulatory action of food intake, hCG and 5-HT4 receptor agonists. In addition, measurement of ACTH concentrations in plasma obtained from two AIMAH patients during adrenal vein sampling showed a significant ACTH gradient versus periphery indicating that AIMAH tissues actually secrete ACTH in vivo. The ACTH receptor antagonists corticostatin and ACTH(7-38) reduced basal as well as GIP-induced cortisol production from perifused hyperplasia tissues. These data indicate that, in AIMAH tissues, ACTH released by a subpopulation of steroidogenic cells exerts an intraadrenal stimulatory tone on cortisol secretion. They also suggest that macronodular bilateral adrenal hyperplasia may be regarded as a cause of ACTH-dependent Cushing[apos]s syndrome due to ectopic expression of corticotropin within the adrenal cortex.[br][br]Sources of Research Support: This work was supported by grants from INSERM, Assistance Publique des Hopitaux de Paris, the COMETE network, the Societe Francaise d[apos]Endocrinologie and Novo Nordisk Laboratory.[br][br]Nothing to Disclose: EL, CD, JY, IB, SR, ZB, LG, PC, AT, FG, SC-M, J-MK, YA, JB, HL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 935 194 1528 SUN-503 PO01-01 Sunday 1325 2012


1322 ENDO12L_SUN-504 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Potential Role of Intraadrenal Mast Cells in the Physiopathology of Aldosterone-Producing Adenoma Celine Duparc, Lucile Moreau, Felipe Golib, Sheerazhed Boulkroun, Arndt Benecke, Francoise Gobet, Tchao Meatchi, Pierre-Francois Plouin, Maria-Christina Zennaro, Estelle Louiset, Herve Lefebvre Institute for Biomedical Research and Innovation, Rouen University, Mont-Saint-Aignan, France; Institute for Biomedical Research and Innovation, University Hospital of Rouen, Rouen, France; Institut des Hautes Etudes Scientifiques (IHES), Bures sur Yvette, France; European Hospital Georges Pompidou, Paris, France; Institute for Biomedical Research and Innovation, University Hospital of Rouen, Rouen, France; European Hospital Georges Pompidou, Paris, France; European Hospital Georges Pompidou, Paris, France We have shown that 5-hydroxytryptamine (5-HT) receptor type 4 (5-HT4R) agonists stimulate aldosterone production in patients with aldosterone-producing adenoma (APA). Moreover, 5-HT-positive cells were observed in APA tissues and 5-HT4R mRNAs were overexpressed in APAs in comparison with normal adrenals. All these results suggested that 5-HT produced by adrenal mast cells could be involved in the physiopathology of APAs. The aim of this work was to investigate the role of mast cells in the physiopathology of APA. Mast cells were characterized by immunohistochemical stainings using antibodies against tryptase. A high density of mast cells was observed in peritumoral adrenal tissue surrounding adenomas or in the tumor tissues. Mast cell-specific gene expression profiling discriminated APA from normal adrenal tissues by use of unsupervised clustering analysis. Tryptase labelling was detected in some steroidogenic cells in the vicinity of mast cells, suggesting interactions between these two cell types. In addition, staining of APA tissues with antibodies to tryptophane hydroxylase, the key enzyme of 5-HT biosynthesis, revealed the presence of immunoreactivity in both mast cells and a subpopulation of steroidogenic cells. In most organs, mast cells establish contacts with sympathetic nervous fibers. Using double immunostaining of APA tissues with tryptase and protein S100, we observed protein S100-positive fibers in peritumoral adrenal tissue as well as interactions between nervous fibers and mast cells suggesting that the secretory activity of mast cells is controlled by neural inputs.[br]Interactions between mast cells and adrenocortical cells were explored using the LAD2 human mast cell line and the human adrenocortical cell line H295R. Administration of LAD2 cell culture supernatant to H295R cells induced a significant increase in aldosterone production which was potentiated when LAD2 cells were pre-incubated with the mast cell degranulator, compound 48/80. On the other hand, we observed that 5-HT stimulated H295R cells steroidogenesis via 5HT4R activation suggesting that the effect of mast cells supernatant could be at least in part due to 5-HT. These studies will be completed by co-cultures of mast cells and adrenocortical cells in the absence and presence of specific 5HT4R antagonists.[br]Altogether, our results show that APA tissues contain numerous mast cells which may influence aldosterone secretion through local release of soluble regulatory factors.[br][br]Sources of Research Support: Grants from the ANR (PHYSIO, ISPA), the FRHTA and the COMETE network (PHRC AOM 06179).[br][br]Nothing to Disclose: CD, LM, FG, SB, AB, FG, TM, P-FP, M-CZ, EL, HL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 944 194 1529 SUN-504 PO01-01 Sunday 1326 2012


1323 ENDO12L_SUN-505 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Removal of the Dominantly Secreting Adrenal Lateralized by Bilateral Adrenal Venous Sampling (BAVS) with Glucagon Stimulation Significantly Alleviated Hypertension in Patients with Bilateral Adrenal Pheochromocytoma Chandy Lou Patiag Malong, Mary Jane Tanchee Ngo, Pilar Torres Salvador, Karel Pacak, Leilani Mercado Asis University of Santo Tomas Hospital, Manila City, Philippines; National Institutes of Health, Bethesda, MD [bold]BACKGROUND:[/bold] The current recommendation on the use of venous sampling is reserved for difficult cases of small pheochromocytoma. Bilateral adrenal venous sampling (BAVS) with glucagon stimulation has been reported to be safe and useful in the early diagnosis of the disease. For bilateral lesions, localization can be challenging but highly important since the removal of the dominant side can markedly improve fatal cardiovascular outcomes as a consequence of chronic hypertension.[br][bold]OBJECTIVE:[/bold]To demonstrate the usefulness of glucagon-stimulated BAVS in determining the dominant adrenal to be removed.[br][bold]METHOD[/bold][bold]OLOGY[/bold][bold]: [/bold]This is a cross-sectional study wherein records of patients who underwent BAVS with glucagon stimulation from 1997-2010 were reviewed.[br][bold]RESULTS:[/bold] Of the 46 patients who underwent BAVS with glucagon stimulation, 19 were diagnosed with bilateral pheochromocytoma. The mean age at diagnosis was 33[plusmn]14 years. The mean duration of hypertension was 5[plusmn]6 years with an average highest systolic blood pressure (BP) of 186[plusmn]30 mmHg and diastolic BP of 113[plusmn]18 mmHg. Headache (68%) is the most common symptom followed by paroxysmal hypertension (58%), palpitation (42%), and flushing (37%).Majority were taking 3 or more anti-hypertensive drugs. On glucagon-stimulated BAVS, 63% had right adrenal dominance. The mean epinephrine and norepinephrine levels on the dominant side were 24,506 [plusmn] 30710 pg/mL and 8,642 [plusmn] 13,395pg/mL, respectively. The mean ratio of the dominant versus non-dominant adrenal for epinephrine and norepinephrine were 3.62 and 4.13, respectively. Three patients, who were hypertensive for 1, 6 and 12 years, underwent removal of the dominant adrenal. On follow-up (mean period=36 months), there was marked improvement in BP control [pre-op vs. post-op: (systolic) 160-240 mmHg vs. 120-150 mmHg; (diastolic) 90-110 mmHg vs. 70-90 mmHg]. There was also a reduction in the number of anti-hypertensive medications (from 3-5 to 2 classes of drugs).[br][bold]CONCLUSION:[/bold] BAVS with glucagon stimulation is a valuable tool in the identification of the dominant adrenal to be removed in patients with bilateral pheochromocytoma to alleviate chronic hypertension and prevent fatal cardiovascular outcomes.[br][br]Nothing to Disclose: CLPM, MJTN, PTS, KP, LMA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 637 194 1530 SUN-505 PO01-01 Sunday 1327 2012


1324 ENDO12L_SUN-506 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Association of Pheochromocytoma with Uncommon Vascular Lesions Sunil K Kota, Siva K Kota, Svs Krishna, Lalit K Meher, Kirtikumar D Modi Medwin Hospital, Hyderabad, India; Central Security Hospital, Riyadh, Saudi Arabia; MKCG Medical College, Berhampur, India Objective:To report associated vascular lesions in pheochromocytoma and discuss possible mechanisms.[br]Methods: From 1990 to 2010,50 patients were diagnosed with pheochromocytoma/paragangliomas.Hospital charts were reviewed retrospectively for coexistent vascular lesions.[br]Results:50 patients (M:F=35:15) with mean age 45.5[plusmn]23.3 years had pheochromocytoma (42 adrenal and 8 extra adrenal).7 patients (14%) had coexisting vascular lesions including renal artery stenosis (RAS) in 4,aortoarteritis in 1,aortic aneurysm in 1 and inferior vena cava(IVC) thrombosis in 1.All had pheochromocytoma [adrenal in 6 patients (4-left,1-right) and ectopic in 1 (at left renal hilum)].RAS was suspected because of small ipsilateral kidney in 2,delayed nephrogram in 1 and impingement of renal artery in 1 patient.A patient with RAS due to intimal fibrosis was offered percutaneous baloon angioplasty,other 3 improved after adrenalectomy and lysis of fibrous adhesive bands.Oral steroids and anticoagulants were given to patients with aortoarteritis and IVC thrombosis.The patient with abdominal aortic aneurysm was advised for annual follow up on account of its size of 4.5 cm and asymptomatic lesion.[br]Discussion:Pheochromocytoma has been described previously in coexistence with RAS,renal artery aneurysm,inferior vena cava thrombosis(1).Though the coexistence can reflect chance association,we propose certain causative factors.Mechanism for RAS in pheochromocytoma:1)tumor compression,2)catecholamine induced vasospasm, 3)periarterial adhesion,4)associated atherosclerosis and fibromuscular dysplasia.Mechanism for associated aortoarteritis:1)catecholamine induced endothelial damage and intimal fibrosis, 2)association with autoimmune conditions, eg SLE,Behcet[apos]s disease.Mechanisms for associated IVC thrombosis:1)local compression leading to stasis 2)sustained hypertension leading to vascular endothelial injury and hypercoagulbility,3)association with SLE, Behcet[apos]s disease and 4)An underlying anatomic abnormality or coagulation disorder.Mechanism for associated abdominal aortic aneurysm:1)persistent exposure to high catecholamines induced vascular wall damage and weakening 2)Associated cigarette smoking, increasing age, hypertension and atherosclerosis 3)Coexistence of vasculitis like takayasu[apos]s disease,Giant cell arteritis and cystic medial necrosis.[br]Conclusion:The state of catecholamine excess and various other coexisting factors can lead to simultaneous occurrence of uncommon vascular abnormalities.[br][br]1. Gill IS, et al., J Urol. 2000; 164: 296.[br][br]Nothing to Disclose: SKK, SKK, SK, LKM, KDM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1408 194 1531 SUN-506 PO01-01 Sunday 1328 2012


1325 ENDO12L_SUN-507 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) Prevalence of Succinate Dehydrogenase Subunit B (SDHB) and D (SDHD) Mutations in Pheochromoctyoma/Paraganglioma in a Single Center in Singapore Dawn Shao Ting Lim, Lih Ming Loh, Yi Zhao, Peng Chin Kek Singapore General Hospital, Singapore, Singapore; Singapore General Hospital, Singapore, Singapore Germline mutations of the genes encoding succinate dehydrogenase subunits B (SDHB) and D (SDHD) predispose to the autosomal dominantly inherited paraganglioma syndromes type 4 (PGL-4) and type 1 (PGL-1) respectively. Malignancy is more frequently seen in SDHB mutation carriers and SDHD mutation carriers more often present with multifocal tumours.[br]7.5-27.0% of tumours without an obvious syndrome or family history result from unsuspected germline mutations. The prevalence of SDHB and SDHD mutations in patients with non-familial tumours has been reported to range from 1.5-10% and 0.8-10% respectively. We report the prevalence of SDHB and SDHD mutations in non-familial pheochromoctyoma/paraganglioma in a single centre in Singapore.[br]16 sporadic cases of pheochromocytoma/paraganglioma diagnosed and followed-up between 1998 and 2011 were studied. All patients underwent genetic analysis for SDHB and SDHD mutations. 12/16 (75%) presented with pheochromocytomas and 4/16 (25%) with paragangliomas. None had a positive family history. 1/16 (6.3%) had a SDHB mutation and 3/16 (18.8%) had SDHD mutations.[br]Metastatic extra-adrenal disease was observed in the single patient with a SDHB mutation, consistent with the increased risk of malignancy associated with these mutations. C/T substitutions were found on exon 6, and C/G substitutions on exon 8 of the SDHB gene.[br]2/3 patients with SDHD mutations had multifocal pheochromocytomas/paragangliomas. One had bilateral dopamine-secreting carotid body paragangliomas, supporting previous reports that SDHD gene mutations account for about 50% of all familial head and neck paragangliomas. Both patients had G/C substitutions in exon 1. The remaining patient had an exon 3c deletion. This patient had a solitary pheochromocytoma and no recurrent disease 12 years following resection, highlighting that advanced age and isolated tumours do not preclude familial disease.[br]The majority of patients without SDHB and SDHD mutations had solitary pheochromocytomas (83.3%) and one had a solitary intra-abdominal paraganglioma with no recurrence 3 years following resection. 1/12 (8.3%) had a metastatic pheochromocytoma, which is higher than the expected 4% malignancy rate of sporadic tumours, but may represent a selection bias.[br][br]Nothing to Disclose: DSTL, LML, YZ, PCK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1525 194 1532 SUN-507 PO01-01 Sunday 1329 2012


1326 ENDO12L_SUN-508 POSTER SESSION: Clinical [amp] Molecular Aspects of Adrenocortical Tumors (1:30 PM-3:30 PM) The Role of Succinate Dehydrogenase Complex Subunit A Mutations in Paraganglioma and Pheochromocytoma Syndromes Julia Katharina Bickmann, Stefanie Sollfrank, Racha Hassoun, Arno Schad, Konstantinos Papaspyrou, Wolf J Mann, Matthias M Weber, Karl J Lackner, Heidi Rossmann, Christian Fottner University Medical Center Mainz, Mainz, Germany; University Medical Center Mainz, Mainz, Germany; University Medical Center Mainz, Mainz, Germany; University Medical Center Mainz, Mainz, Germany [bold]Objective[/bold]: SDHA (5p15 MIM ID *600857) encodes subunit A, a flavoprotein constituting the catalytic domain of the Succinate Dehydrogenase (SDH) complex. We sought to investigate whether SDHA mutations known to occur homozygously in Leigh Syndrome (MIM #256000) could have any effect on the development of paragangliomas in our patient collective.[br][bold]Design and Methods[/bold]: We designed sequencing assays of all 15 exons of the SDHA gene (NT_023089) including their splice sites. The SDHA genes of 72 pheochromocytoma and paraganglioma index patients hitherto tested negative for mutations in SDHB, SDHC, SDHD, TMEM127, MAX and the RET-protocongene and VHL were then systematically sequenced. Pyrosequencing was applied in estimating the frequencies of potential SNPs and mutations of inexplicit pathogenicity in a healthy control population.[br][bold]Results and Conclusions[/bold]: One patient (43 years, female) affected by a glomus jugulare tumor carried a nonsynonymous substitution in codon 1 of the SDHA gene (c.1ATG[gt]GTG, P.MET[gt]VAL). According to Polyphen (http://genetics.bwh.harvard.edu/pph2) and SIFT (http://sift.jcvi.org/) a deleterious affection of the resulting protein is very likely. Pyrosequencing confirmed that the c.1A[gt]G mutation is unique in our own control population.[br]Western Blot analysis demonstrated a lower SDHA protein concentration than in healthy control subjects and an additional band at 42 kDa. SDHA and SDHB immunohistochemistry in the patient[apos]s tumour sample will be applied to confirm a disruption of the SDH complex. For the second time a heterozygous SDHA mutation is reported in a patient affected by paraganglioma (Burnichon et al., 2010). In the future SDHA should be considered in genetic testing of paraganglioma patients who have undergone tumor surgery particularly if the testing of SDHD, SDHB and SDHC has failed to identify the underlying mutations.[br][br]Nothing to Disclose: JKB, SS, RH, AS, KP, WJM, MMW, KJL, HR, CF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1044 194 1533 SUN-508 PO01-01 Sunday 1330 2012


1345 ENDO12L_SUN-535 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) Autophagy Is Required for Androgen Receptor-Mediated Prostate Cell Growth and Survival Yan Shi, Jayantha B Tennakoon, Jenny J Han, Fatima A Merchant, Matthew K Howe, Olga R Ilkayeva, Christopher B Newgard, Donald P McDonnell, Daniel E Frigo University of Houston, Houston, TX; University of Houston, Houston, TX; Duke University, Durham, NC Androgens regulate both the physiological development of the prostate and the pathology of prostatic diseases. Despite these known androgenic roles, it is unclear which specific cellular processes govern these biologies and exactly how these processes impact cellular energy demands. Our data indicate that androgens regulate overall cell metabolism and thus cell growth, in part, by increasing autophagy in prostate epithelial cells. Importantly, inhibition of autophagy using either pharmacological or molecular inhibitors significantly abrogated androgen-induced prostate cancer cell growth. Mechanistically, androgen-mediated autophagy appears to promote cell growth and survival by breaking down and redirecting intracellular fats, a process known as lipophagy, for use in [beta]-oxidation and subsequent ATP synthesis. These findings, combined with previous studies demonstrating roles for increased fat metabolism and autophagy in prostatic neoplasias, highlight the potential of targeting underexplored metabolic pathways for the development of novel therapeutics.[br][br]Nothing to Disclose: YS, JBT, JJH, FAM, MKH, ORI, CBN, DPM, DEF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1617 195 1552 SUN-535 PO26-01 Sunday 1349 2012


1346 ENDO12L_SUN-536 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) Multi-Parametric, Single-Cell High-Content Analysis To Classify Androgen Receptor Effectors in Prostate and Breast Cancer Cells Justin Y Newberg, Fabio Stossi, Radhika D Dandekar, Michael M Mancini Baylor College of Medicine, Houston, TX High content analysis (HCA) is increasingly recognized as a powerful tool to quantitatively analyze single cell responses to compounds, with image- and statistics-based tools being used to classify effectors based upon multiple mechanistic and cytological parameters. Here, we present HCA-based characterization of compounds in terms of androgen receptor (AR) action in prostate and breast cancer cell lines by measurement of cell-by-cell determination of: 1) AR protein levels; 2) nuclear translocation; 3) nuclear hyperspeckling; 4) subpopulation analysis; and, 5) application of [gt]100 additional descriptive features obtained from an image-based informatics approach.[br]Enabled by high-throughput fluorescence microscopy, we measured immunoreactive AR via HCA in androgen sensitive (LNCaP) and castrate resistant prostate cancer cells (LNCaP C4-2 and 22Rv1); and in AR+ breast cancer cells (MCF-7). Initially, we treated cells with a panel of known AR agonists and antagonists (e.g., DHT, OHF, MDV3100, bicalutamide, nilutamide) in a dose, combination and time-dependent manner and used HCA to extract which features were more accurate in describing the character of agonists vs. antagonists. Statistical methods were then used to identify changes in AR responsiveness (e.g., receptor stabilization, nuclear translocation, hyperspeckling) as a function of the compound treatments. Classification analysis revealed that nilutamide treatments exhibited cell-specific antagonism in LNCaP versus C4-2 cell lines (with stronger antagonism in C4-2). Next, given the proposed endocrine disrupting activities of BPA, we expanded our analysis to determine if BPA or BPA analogs influence AR metrics. Interestingly, preliminary results indicate that DMB BisA also reduced AR levels. Having established an automated methodology for identifying differences in AR responses at the single-cell level across multiple cell lines, we are now expanding analysis to a larger group of endocrine disruptors and other compound classes to further identify differences between cell lines and to assess image-based multi-parametric analysis in drug and endocrine disruptor screening.[br][br]Nothing to Disclose: JYN, FS, RDD, MMM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2201 195 1553 SUN-536 PO26-01 Sunday 1350 2012


1347 ENDO12L_SUN-537 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) FoxA1 Plays an Important Role in Specifyng Chromatin-Binding Sites Unique to the Androgen Receptor Biswajyoti Sahu, Marko Laakso, Kristian Ovaska, Ievgenii Sinielnikov, Sampsa Hautaniemi, Olli A Janne Biomedicum Helsinki, University of Helsinki, Helsinki, Finland; Biomedicum Helsinki, University of Helsinki, Helsinki, Finland Androgen receptor (AR) and glucocorticoid receptor (GR) are ligand-inducible transcription factors that belong to the superfamily of nuclear receptors. These receptors bind their cognate ligands and mediate physiological and pathophysiological actions of androgens and glucocorticoids. Recent genome-wide studies have shown that AR and GR bind to chromatin [italic] in vivo [/italic] far away from transcription start sites of their target genes, implying a distal regulatory mode of transcriptional control. Comparison of AR cistromes in two prostate cancer cell lines (LNCaP-1F5 and VCaP cells) revealed that over 85% of the AR cistrome in LNCaP-1F5 cells overlaps with that in VCaP cells. AR-binding sites (ARBs) unique to LNCaP-1F5 cells require not only the presence of the [italic] cis[/italic]-element for forkhead proteins but also prior binding of FoxA1 to these sites. Depletion of FoxA1 in LNCaP-1F5 cells abrogates AR binding, implying that AR binds to these unique chromatin sites through tethering to FoxA1. By contrast, FoxA1 fails to bind to the corresponding loci in VCaP cells explaining the lack of the ARBs at these sites. FoxA1 protein levels in the two cell lines are very similar. LNCaP-1F5 cells express also glucocorticoid receptor (GR), permitting analyses of AR and GR cistromes as well as androgen- and glucocorticoid-dependent transcription programs in a single cell type. Comparison of AR and GR cistromes in LNCaP-1F5 cell chromatin revealed that there are receptor binding sites that are unique to either AR or GR, as well as shared by both AR and GR; 57% of the AR cistrome overlaps with that of GR. Androgen and glucocorticoid-regulated transcription programs were commensurate with receptor binding site distribution. Importantly, the [italic] cis[/italic]-element that specifies chromatin binding sites unique to AR is not a classical androgen response element but a composite [italic] cis[/italic]-element (1) that includes FoxA1-binding motif. Collectively, our results indicate that FoxA1 plays an important role in assuring AR binding specificity among prostate cancer cell lines as well as between two nuclear receptors, AR and GR.[br][br]1. Sahu et al., EMBO J 2011; 30:3962.[br][br]Nothing to Disclose: BS, ML, KO, IS, SH, OAJ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1402 195 1554 SUN-537 PO26-01 Sunday 1351 2012


1348 ENDO12L_SUN-538 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) Transcriptional Regulation of FoxO3 Gene by Glucocorticoids Taiyi Kuo, Patty H Liu, Jenny New, Jen-Chywan Wang University of California, Berkeley, CA Glucocorticoids and Foxo3 exert similar metabolic effects in skeletal muscle. Foxo3 gene expression was increased by dexamethasone (Dex), a synthetic glucocorticoid, in mouse C2C12 myotubes and gastrocnemius muscle. In C2C12 myotubes the increased expression is due to, at least in part, the elevated rate of Foxo3 gene transcription. In the mouse Foxo3 gene we identified three glucocorticoid receptor (GR) binding regions (GBR): one in upstream of the transcription start site, -17kbGBR; and two in introns, +45kbGBR and +71kbGBR Together, these three GBR contain 4 glucocorticoid response elements (GRE). Micrococcal nuclease (MNase) assay showed that 30 min Dex treatment increased the sensitivity to MNase in the GRE of +45kbGBR, but not in +71kbGBR. Upon 60 min Dex treatment; however, the sensitivity to MNase was elevated in +71kbGBR, but not in +45kbGBR. Dex treatment for 30 and 60 min did not affect the chromatin structure of the -17kbGBR whose GRE was located in a linker region, whereas the GRE of +45kbGBR and +71kbGBR were located on nucleosomes. Dex treatment for 30 min also increased the levels of acetylated histone H3 and H4 in all three GBR. Finally, using chromatin conformation capture assay, we showed that Dex treatment increased the interaction between the -17kbGBR and two genomic regions: one located around +500 bp and the other located around +73 kb. Overall, our results indicate that glucocorticoids activated Foxo3 gene transcription through multiple GRE. Each GRE was regulated by distinct mechanisms, and DNA looping is likely involved in this transcriptional activation process.[br][br]Sources of Research Support: NIH (R01DK083591); Muscular Dystrophy Association (186068); Dissertation Award Fellowship from the University of California Tobacco-Related Diseases Research Program awarded to TK.[br][br]Nothing to Disclose: TK, PHL, JN, J-CW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 739 195 1555 SUN-538 PO26-01 Sunday 1352 2012


1349 ENDO12L_SUN-539 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) The Structure and Functions of the Glucocorticoid Receptor[apos]s Intrinsically Disordered Activation Function-1 (AF1) Domain Shagufta H Khan, Raj Kumar The Commonwealth Medical College, Scranton, PA Most of our understanding of the two transcriptional activation (AF) domains (AF1 and AF2) of nuclear receptors (NRs) is with AF2 that is composed of a hydrophobic pocket within the globular C-terminal ligand binding domain (LBD). In response to conformational changes induced by hormone, AF2 recognizes LXXLL motifs of proteins such as the p160 family of steroid receptor coactivators (SRC-1,-2,-3). Despite the fact that AF1, located in the N-terminal domain (NTD), constitutes the major transcriptional activity of steroid receptors (SRs), a subfamily of NRs, little is known about this domain including its structure, the biologically relevant interacting coregulators and mechanism of activation. This deficiency is due to the fact that the NTD/AF1 is composed largely of intrinsically disordered polypeptide (IDP) and is not amenable to traditional structure analysis by X-ray crystallography. IDPs undergo a disorder-order transition upon interaction with target molecules by a [ldquo]coupled folding and binding process[rdquo] that can result in stabilization of an active structural conformation. However, the molecular and structural basis for these interactions and mechanisms of AF1 activation remains poorly defined. Our biophysical/structural data show that when incubated in the presence of a naturally occurring compound, trehalose, AF1 of the glucocorticoid receptor (GR) adopts a significantly higher helical structure with characteristics of native-like functional conformation. We further tested whether interaction of an AF1 binding protein (BP) also induces such structure formation in AF1. Our data show that BP binding to AF1 induces more helical content in AF1 at the expense of random coil conformation, similar to that induced by trehalose. Further, protein:protein interaction data show that in this folded conformation, AF1[apos]s interaction with specific coactivators. These results suggest that under physiological conditions, AF1 may be adopting such structures for its efficient interactions with specific coregulators. Our results provide a mechanism by which SRs[apos] structural dynamics within the NTD/AF1 plays a critical role in regulating transcriptional activity of AF1 through disorder-order transitions induced by interactions with critical coregulators. A better understanding of how structural dynamics of NTD/AF1 mediates cell/tissue and target gene specific responses to SRs is essential for improved therapeutic targeting of SRs.[br][br]Nothing to Disclose: SHK, RK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1253 195 1556 SUN-539 PO26-01 Sunday 1353 2012


1350 ENDO12L_SUN-540 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) Methylglyoxal Induces ARPE-19 Cell Death Via Downregulation of GR Function Seung-Jin Kim, Sungsoo Park, Hojung Choi, Jin Sook Kim, Young Sook Kim, Eungseok Kim College of Natural Sciences, Chonnam National University, Gwangju, Republic of Korea; Korea Institute of Oriental Medicine (KIOM), Daejeon, Republic of Korea Diabetic retinopathy is a major complication of diabetes and breakdown of blood-retinal barrier (BRB) leads to vascular leakage which eventually results in diabetic macular edema. The outer BRB is composed of RPE cells which play an important role in the fluid homeostasis of neural retina. In addition, RPE cells secrete a variety of factors such as VEGF and PEDF which are essential for maintenance of inner retinal integrity. Therefore, apoptosis of RPE cells induced by MGO which are elevated in diabetic condition cause a deleterious effect on the retina, leading to the pathogenesis of diabetic retinopathy.[br]We demonstrate here that GR can inhibit MGO-induced RPE cell death. RT-PCR analysis clearly showed that MGO inhibited GR mRNA levels in RPE cells in a dose-dependent manner. Suppressive effect of MGO on GR expression was further confirmed by the reporter gene assay in which MGO strongly repressed GR promoter activity. In addition, this inhibitory effect of MGO on GR mRNA levels was partially reduced by addition of JNK or p38 inhibitor. Furthermore, MGO also inhibited GR transcriptional activity in a dose-dependent manner, suggesting that MGO is able to inhibit GR signaling through regulation of not only GR expression but also GR activity. MGO treatment also induced RPE cell death and addition of RU486, a GR antagonist, further enhanced MGO-induced RPE cell death. Consistent with cell death assay, MGO treatment increased cleavage of PARP in RPE cells while MGO reduced expression of the cell survival genes such as Bcl-2 and Bcl-xL. However, when Dexamethasone, a GR agonist, was treated together with MGO, cleaved PARP was decreased as compared with that of MGO-treated RPE cells. In addition, Dex also inhibited MGO-induced suppression of Bcl-2 and Bcl-xL expression.[br]These findings indicate that MGO inhibits GR expression and activity which results in decrease of survival gene expression, leading to RPE cell death.[br][br]Sources of Research Support: This research was supported by grant K11040 from Korea Institute of Oriental Medicine.[br][br]Nothing to Disclose: S-JK, SP, HC, JSK, YSK, EK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 595 195 1557 SUN-540 PO26-01 Sunday 1354 2012


1351 ENDO12L_SUN-541 POSTER SESSION: Roles of Androgen [amp] Glucocorticoid Receptors in Physiology [amp] Disease (1:30 PM-3:30 PM) Transcriptomine: The Nuclear Receptor Signaling Transcriptome Database Scott A Ochsner, Christopher M Watkins, Apollo McOwiti, David L Steffen, Lauren B Becnel, Neil J McKenna Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX Understanding signaling by nuclear receptors (NRs) requires an appreciation of their cognate ligand- and tissue-specific transcriptomes. While target gene regulation data are abundant in this field, they reside in hundreds of discrete publications in formats refractory to routine query and analysis and, accordingly, their full value to the NR signaling community has not been realized. One of the mandates of the Nuclear Receptor Signaling Atlas (NURSA) is to facilitate access of the community to existing public datasets. Pursuant to this mandate we are developing a freely-accessible community web resource, Transcriptomine, that for the first time bring together the sum total of available expression array data points generated by the field in a single location at the NURSA website, www.nursa.org. Transcriptomine currently contains approximately 12 million gene fold change data points within 700 contrasts covering 21 NRs, 54 NR ligands, 10 NR coregulators, and 93 specific tissues/cell lines. Transcriptomine is designed to accommodate a spectrum of end users ranging from the bench researcher to those with advanced bioinformatic training. We show how Transcriptomine was mined for all-trans retinoic acid regulated gene candidates in a variety of cell lines which were subsequently verified via PCR in mouse ES cells. Our resource affords an entirely new paradigm for leveraging gene expression data in the NR signaling field and will, when complete, empower users to assemble complex queries that span diverse molecules, tissues [amp] cell lines, target genes, biological functions and disease associations, and that would otherwise be prohibitive in terms of time and effort. Transcriptomine will be regularly updated with gene lists from future genome-wide expression array and expression-sequencing datasets in the NR signaling field.[br][br]Sources of Research Support: NIH Grant U19DK62434.[br][br]Nothing to Disclose: SAO, CMW, AM, DLS, LBB, NJM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2199 195 1558 SUN-541 PO26-01 Sunday 1355 2012


1352 ENDO12L_SUN-542 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Single Cell-Based Image Analysis of Nuclear Receptor/Coregulator Functions in Breast Cancer Cells Reveals Heterogeneous Subpopulations: Development of Biological Response Fingerprints for Ligands and Endocrine Disruptors Fabio Stossi, Justin Y Newberg, Radhika D Dandekar, John K Westwick, Cheryl L Walker, Michael A Mancini Baylor College of Medicine, Houston, TX; Odyssey Thera, San Ramon, CA; Texas A[amp]M Health Science Center, Houston, TX Modern microscopy- and flow cytometry-based analyses indicate tumor cell populations can be markedly heterogeneous. Yet, the vast majority of approaches used to investigate tumorigenic mechanisms and therapeutic responses are based upon single-point assays of large populations of cells, where subpopulation responses are undetectable. For example, whereas breast cancers are generally classified as either estrogen receptor-alpha positive (ER+) or negative (ER-), in ER+ tumors there is a broad range of ER levels in individual cells. Despite a growing awareness of the importance of tumor cell heterogeneity and outlier biology in cancer therapeutics, there is a paucity of data on the potential interplay between multiple nuclear receptors and coregulators, their heterogeneity of expression and functions, and their combinatorial role in therapeutic response, or in sensing environmental stressors (e.g., endocrine disruptors).[br]To specifically quantify biological responses on an individual cell basis rather than averaged responses from bulk populations, we initially performed single cell-based high content analysis (HCA) to examine MCF-7 cells in terms of expression of several type-1 nuclear receptors, and responsiveness to individual or mixed compound treatments (agonist, antagonists, endocrine disruptors). Quantitative immunofluorescence reveals a wide range of heterogeneity (up to log scale differences) in the expression and ratios of ER, androgen and glucocorticoid receptors (AR, GR). Multiparametric analyses describing NR expression and spatial distribution were used to generate a matrix that quantitatively classifies subpopulations of MCF-7 cells. Next, we expanded our analysis to selected coregulators (e.g., SRC-3), with endogenous target gene expression via mRNA fluorescence in situ hybridization now in progress. Statistical analysis of the subpopulations datasets facilitated creation of [ldquo]biological response fingerprints,[rdquo] which were used to classify the effect of individual or mixed ligand treatments. We are expanding our HCA approach to other breast cancer cell lines and are including additional metrics of cell cycle and receptor-coregulator interactions by protein complementation assays. Collectively, these high throughput-amenable approaches are poised to greatly aid in the understanding the role of heterogeneity in breast cancer, and serve as a robust model for next-generation drug screening and endocrine disruptor assays.[br][br]Nothing to Disclose: FS, JYN, RDD, JKW, CLW, MAM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2259 196 1559 SUN-542 PO26-02 Sunday 1356 2012


1353 ENDO12L_SUN-543 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Chromatin Interplay between ER[alpha] and GR Fine-Tunes Transcriptional Output Michael James Bolt, Fabio Stossi, Justin Newberg, Michael Mancini Baylor College of Medicine, Houston, TX Type 1 nuclear receptors (NR) are key transcription factors that regulate their target genes via binding to specific DNA steroid response elements (SREs), and followed by recruiting coactivators and histone modifying proteins that collectively modify RNA Polymerase II activity. In this study we utilized a microscopically-visible estrogen receptor alpha (ER[alpha]) regulated transcription array (1)(2)(3) to analyze the interplay between type I NRs. We show a coordinated loading onto the array of ER[alpha] and other steroid receptors only in the presence of dual agonist treatments (e.g., estradiol (E2) + dexamethasone (Dex), E2 + dihydroxytestosterone (DHT), etc.). We then focused on the ER[alpha]/GR pair and showed that the cooperative loading phenomenon is maximal when both receptors are agonist bound, and that GR loading is dependent upon ER[alpha] both occupancy on the array and DNA binding. Moreover, use of specific ER[alpha] mutants reveals helix-12 is required for GR array loading. We next demonstrate that the coordinated receptor interplay also occurs in MCF-7. For example, addition E2 + Dex decreases ER[alpha] recruitment at the TFF1/pS2 promoter while increasing GR promoter loading. This scenario causes an overall negative effect on the TFF1/pS2 mRNA compared to the E2 alone treatment. However, in terms of ER[alpha] and GR recruitment, we also identify genes that show different combinations of outcomes. In conclusion, both our single cell- and biochemistry-based approaches identify cooperative coordination of the actions of ER[alpha] and Type I NRs on chromatin. This interrelationship warrants further investigation through genome-wide studies, which we hypothesize will reveal a gene specific interplay between different NRs.[br][br](1) Sharp et al, 2006, J Cell Sci. (2) Berno et al, 2008, PloSOne. (3) Ashcroft et al, 2011, Gene.[br][br]Sources of Research Support: National Library of Medicine Keck Center for Interdisciplinary Bioscience Training of the Gulf Coast Consortia (NLM Grant No. T15 LM007093).[br][br]Nothing to Disclose: MJB, FS, JN, MM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1423 196 1560 SUN-543 PO26-02 Sunday 1357 2012


1354 ENDO12L_SUN-544 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Genomics and Integration of Estrogen Receptor Alpha and Protein Kinase Chromatin Binding and Regulation of Gene Expression and Cell Properties with a Tissue-Selective Estrogen Complex (TSEC) Zeynep Madak-Erdogan, Ping Gong, Benita S Katzenellenbogen University of Illinois and College of Medicine at Urbana-Champaign, Urbana, IL Estrogens act through estrogen receptors (ERs) in the nucleus of target cells to regulate gene expression, but they also act through ERs at extranuclear sites, where they activate kinase cascades. Recent work (e.g., Madak-Erdogan et al., Mol. Cell. Biol. 31:226, 2011) has revealed that estrogen action in target cells requires genomic collaboration between ERs and protein kinases in the control of gene expression and cell phenotypic properties. Therefore, in the current study, we have investigated the actions of estrogens, selective estrogen receptor modulators (SERMs), and a Tissue-Selective Estrogen Complex (TSEC), and the involvement of nuclear and extranuclear pathways in their activities. Chromatin immunoprecipitation (ChIP) and gene expression analyses were utilized to monitor the recruitment of ER[alpha] and the extracellular signal-regulated kinase 2 (ERK2) to ER binding sites in chromatin of estrogen-regulated genes, and these were correlated with changes in cell phenotypic properties.[br]We found that conjugated estrogen (CE, which is a mix of 10 estrogens as present in Premarin[sup][reg][/sup]) was much less effective than estradiol (E2) in ERK2 recruitment to the same chromatin binding sites, whereas recruitment of ER[alpha] to chromatin binding sites by E2 and CE was relatively similar, although lower for CE at certain sites. Likewise, MAPK activation was lower for CE vs E2. CE was also much less agonistic than E2 in stimulation of the proliferation of MCF-7 breast cancer cells. The SERM bazedoxifene (BZA, 1 [micro]M) fully suppressed any cell proliferation stimulated by E2 or CE (at 1 pM [ndash] 1 [micro]M), and it reversed gene stimulation by CE or E2, with BZA being as or more suppressive than Faslodex. The findings suggest that the balance of biological activities mediated through direct nuclear vs. kinase-mediated pathways is different for CE vs. E2, with CE having less potency in the kinase-mediated pathways. These studies provide a molecular underpinning for the basis by which a TSEC functions and is most effective. They also expand our understanding of the mechanisms and different modes by which estrogens and SERMs support or antagonize one another in regulating chromatin binding, gene expression, and cell phenotypic properties, such as proliferation, and they illuminate how the combined actions of these two classes of hormones (as present in TSECs) can achieve unique modes of regulation and efficacy.[br][br]Sources of Research Support: Investigator Initiated Research (IIR) grant from Pfizer.[br][br]Disclosures: BSK: Principal Investigator, Pfizer, Inc. Nothing to Disclose: ZM-E, PG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 780 196 1561 SUN-544 PO26-02 Sunday 1358 2012


1355 ENDO12L_SUN-545 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Botanical Estrogens: Molecular Mechanisms and Cellular Pathways of Activity in Target Cells Yan Jiang, Methu Jeyakumar, Kathryn Carlson, Ikhlas Khan, William G Helferich, John A Katzenellenbogen, Benita S Katzenellenbogen University of Illinois at Urbana-Champaign, Urbana, IL; University of Illinois at Urbana-Champaign, Urbana, IL; University of Illinois at Urbana-Champaign, Urbana, IL; University of Mississippi, Oxford, MS Botanical estrogens (BEs) are widely consumed by women with the expectation that they provide a safe, natural source of estrogens to replace the loss of endogenous estrogen in the menopause. Because estrogens can have diverse effects in target cells, including the stimulation of some breast cancers, this unregulated consumption of BEs might not contribute uniformly to healthy aging. Estrogens regulate gene transcriptional programs and the physiology of many reproductive and non-reproductive tissues, and the mechanisms by which they act are multi-faceted, involving two estrogen receptors, ER[alpha] and ER[beta], and nuclear-initiated and extranuclear-initiated pathways. With BEs, little is known about these mechanistic and cellular aspects. We are using a systems biology approach to provide a molecular and cellular profile of the activity of BEs, to enable determination of whether they have activities similar to or distinct from estradiol. We have examined molecular interactions of BEs with ER[alpha] and ER[beta]: ligand binding, conformation and dynamics of the ligand-receptor complexes, and interaction with key coregulators. We are also investigating on a genome-wide basis, how the patterns of ER and coregulator recruitment to chromatin binding sites (cistromes) and changes in gene expression (transcriptomes) in target cells are regulated by BEs through nuclear and extranuclear pathways; and we are developing mechanistic systems biology profiles that relate the cistromes, transcriptomes, and pathway-specific actions of BEs to their effects on cell functional properties. In general, we have found that BEs show preference in binding and coactivator recruitment to ER[beta] compared to ER[alpha]. The relative levels of ER[alpha] and ER[beta] in cells therefore greatly impact gene expression and proliferation response to different BEs. Among the BEs tested, potency and efficacy of liquiritigenin and S-equol through ER[beta] were greater than expected from their ER[beta] binding affinities. ER[beta] dampened the pro-proliferative activity of estrogens through ER[alpha], and BEs varied in their effectiveness in suppressing proliferation through ER[beta]. Based on these molecular and cellular profiles, we are able to place cell response to BEs on a mechanistic basis that will allow determination of whether BEs have similar or unique activities compared to those of other estrogens and provide fundamental information to inform future clinical studies.[br][br]Sources of Research Support: NIH [1 P50 AT006268 (BSK, JAK, WGH, IK) from the National Center for Complementary and Alternative Medicines (NCCAM), the Office of Dietary Supplements (ODS) and the National Cancer Institute (NCI)].[br][br]Nothing to Disclose: YJ, MJ, KC, IK, WGH, JAK, BSK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 858 196 1562 SUN-545 PO26-02 Sunday 1359 2012


1356 ENDO12L_SUN-546 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) SUMOylation Contributes to Full Antiestrogenicity Khalid Hilmi, Nader Hussein, Rodrigo Mendoza- Sanchez, Mohamad El Ezzy, Chantal Durette, Houssam Ismail, Justyna Kulpa, Xavier Mascle, Muriel Aubry, Michel Bouvier, Pierre Thibault, Jim Gleason, Sylvie Mader Universit[eacute] de Montr[eacute]al, Montr[eacute]al, Canada; Universit[eacute] de Montr[eacute]al, Montr[eacute]al, Canada; Universit[eacute] de Montr[eacute]al, Montr[eacute]al, Canada; McGill University, Montr[eacute]al, Canada Approximately 70% of breast tumors express or overexpress estrogen receptor alpha (ERa) and are treated with antiestrogens (AEs), which act as competitive inhibitors of this receptor. Tamoxifen (Tam) has been widely used for the treatment of ERa-positive tumors, but intrinsic or acquired resistance can lead to tumor recurrence. Full AEs such as Fulvestrant (Faslodex) are currently used in estrogen receptor alpha (ERa) positive breast cancers in postmenopausal women with disease progression following Tam therapy. Unlike Tam and other Selective estrogen receptor modulators (SERMs), full AEs are devoid of uterotrophic activity in animal models and fully repress the transcriptional activity of ERa in cell models for the partial agonist activity of AEs such as the HepG2 hepatocarcinoma cell line. It is currently thought that the capacity of full AEs to induce rapid polyubiquitination and degradation of ERa underlies their complete suppression of ERa signalling.[br]Here we identify ER[alpha] as a target for Small Ubiquitin-like Modifier (SUMO) posttranslational modification by SUMO1 and SUMO2/3 specifically when liganded with full AEs. Induction of SUMOylation is rapid and precedes receptor degradation in ER-positive breast cancer cells. On the other hand, the SERMs Tam and RU 39 411 do not induce SUMOylation. Deletion mapping analysis indicates that the DNA binding domain (DBD) and helix H12 in the ligand binding domain of ERa are required for full AE-induced SUMOylation. Using mass spectrometry, we identify K171, K180, K299 and K472 as acceptors for SUMO modification. Structure activity relationship experiments with full AE derivatives showed that the induction of SUMOylation is correlated with the degree of inhibition of ER[alpha]-mediated transcription in HepG2 cells. In addition, preventing SUMOylation by overexpression of a SENP deSUMOylase abolished the inverse agonist properties of full AEs without increasing activity in the presence of agonists or of Tam. Together, our results indicate that ERa SUMOylation is a characteristic property of full antiestrogens that contributes to their specific activity profile.[br][br]Nothing to Disclose: KH, NH, RM-S, MEE, CD, HI, JK, XM, MA, MB, PT, JG, SM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 773 196 1563 SUN-546 PO26-02 Sunday 1360 2012


1357 ENDO12L_SUN-547 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Estrogen-Induced Expression of Apurinic Endonuclease 1 (Ape1) Helps Confer Neuroprotection Alicia K Dietrich, Ann M Nardulli University of Illinois, Urbana-Champaign, Urbana, IL Stroke is the third leading cause of death and a major source of permanent disability in the United States. The fact that postmenopausal women are at greater risk of suffering a stroke than premenopausal women suggests that circulating ovarian hormones may be required to reduce the risk of stroke and maintain optimal brain cell function. In support of this idea, a number of studies have shown that 17[beta]-estradiol (E[sub]2[/sub]) attenuates ischemia-induced damage in rodent models of stroke. Interestingly, our laboratory recently showed that E[sub]2[/sub] increases expression of specific oxidative stress response proteins in cultured mammary cells and other laboratories have shown that overexpression of oxidative stress response proteins decreases ischemia-induced brain injury. Thus, we hypothesized that E[sub]2[/sub] may in part mediate its protective effects in the brain by increasing oxidative stress response protein expression.[br]Our studies focused on the oxidative stress response protein apurinic endonuclease 1 (Ape1), which is also known as redox factor-1. Ape1 is a multifunctional protein that, in addition to reducing a number of transcription factors, is also required for DNA repair. Importantly, overexpression of Ape1 in rodents reduces DNA fragmentation and infarct volume in the cerebral cortex after an ischemic event.[br]To determine whether E[sub]2[/sub] altered Ape1 expression in the cerebral cortex, we utilized mouse brain slice cultures, which maintain many of the architectural features and cellular networks found in the intact brain, but permit a more direct assessment of the effects of E[sub]2[/sub] on the brain than is possible using intact animals. Immunofluorescent studies demonstrated that estrogen receptor [alpha] and Ape1 were expressed in the nuclei of cortical neurons and that E[sub]2[/sub] treatment of brain slice cultures increased Ape1 expression in the cerebral cortex. Additionally, quantitative Western blot analysis using extracts from brain slice cultures confirmed that E[sub]2[/sub] increased Ape1 expression and reduced protein damage when oxidative stress was induced. However, when primary cortical neurons were treated with E[sub]2[/sub], no Ape1 induction was observed suggesting that a more robust network of cellular communication is needed for E[sub]2[/sub]-induced Ape1 expression. Our studies provide evidence that E[sub]2[/sub]-induced expression of Ape1 helps to protect the cerebral cortex from an ischemic insult and defines a novel mechanism by which E[sub]2[/sub] may mediate its neuroprotective effects.[br][br]Sources of Research Support: NIH grant R01 DK 053884 to AMN. AKD was supported by NIEHS grant #T32 ES007326.[br][br]Nothing to Disclose: AKD, AMN 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1993 196 1564 SUN-547 PO26-02 Sunday 1361 2012


1358 ENDO12L_SUN-548 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Antagonist Reversal Activity of ER[alpha] L543A, L544A Mutant Correlates with the Ligand-Dependent LBD Dimerization Yukitomo Arao, Laurel A Coons, Kenneth S Korach National Institute of Environmental Health Sciences, RTP, NC; Duke University Medical Center, Durham, NC Estrogen regulates the physiological responses in target cells by means of intracellular estrogen receptors (ERs). Major estrogenic activity appears through the nuclear ER[alpha] and [beta] that activate transcription directly as ligand-dependent transcription factors. ER has two transcriptional activation domains, AF-1 and AF-2. AF-1 is localized in N-terminal region and AF-2 is distributed in the C-terminal ligand binding domain (LBD) of the ER protein. The activity of AF-2 is controlled by the ligands. Namely, helix12 in the LBD constitutes the AF-2 and the configuration of helix12 is changed by agonist or antagonist to the active or inactive form respectively. Previously, we demonstrated that the ER[alpha] mutant of L543A, L544A in helix12 (AF2ER) disrupts AF-2 function and reverses an antagonist, such as ICI182780 (ICI) and tamoxifen (Tam) into an agonist in vitro and in vivo (Arao et al., 2011). N-terminal dependent transactivation function (AF-1) is required for the antagonist reversal activity. However it is still unclear how the antagonist activates the AF-1 of the AF2ER mutant. To understand the molecular mechanism of antagonist reversal activity for AF2ER, we analyzed the relation between the ERE-mediated transcriptional activity and the efficiency of ligand dependent LBD dimer formation using a luciferase reporter assay and a mammalian two-hybrid assay respectively. We found that AF2ER mediated transcription and LBD dimer formation were induced by ICI but not E2; on the other hand, E2 but not ICI induced ER[alpha] WT transcription and LBD dimer formation. The helix12-deleted mutant (LBD/[Delta]H12) induced the ICI-dependent dimer formation but not a F-domain truncated AF2ER mutant (AF2ER-LBD/[Delta]F). ICI-dependent transcription activity of AF2ER[Delta]F was significantly lower than AF2ER. These results suggested that AF2ER mutation dislocated the helix12 and F-domain to induce the antagonist dependent AF2ER-LBD dimer formation and that ligand-dependent dimer formation caused the antagonist reversal activity of AF2ER. We analyzed the antagonist reversal activity of Tam metabolites, 4-hydorxy-Tam (4OHT) and endoxifen (Endox). Endox and 4OHT induced AF2ER mediated transactivation and AF2ER-LBD dimer formation. Both effects of Endox were stronger than 4OHT, supporting that ligand-dependent dimer formation efficiency correlates with the antagonist reversal activity.[br][br]Arao et al., PNAS 2011; 108:14986.[br][br]Sources of Research Support: NIH-Z01ES7006.[br][br]Nothing to Disclose: YA, LAC, KSK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1272 196 1565 SUN-548 PO26-02 Sunday 1362 2012


1359 ENDO12L_SUN-549 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Estrogen Receptor [alpha] Determines Mesenchymal Stem Cell Fate Korinna Wend, Peter Andre Wend, Susan Amy Krum UCLA, Los Angeles, CA; UCLA, Los Angeles, CA In postmenopausal women estrogen levels decline and the bone mineral density decreases. The composition of bone marrow also shifts to contain more adipocytes.[br]To gain deeper insights into the in vivo role of estrogen (E2) during differentiation of bone marrow mesenchymal stem cells we utilized estrogen receptor knock out-mice (ERaKO and ERbKO). The bone marrow from ERaKO mice showed a higher lipid content and increased potential for colony unit formation compared to wild-type or ERaKO animals. Differentiation of ERaKO cells in vitro led to higher amounts of adipocytes but fewer osteoblasts compared to wild-type or ERaKO cells, which suggests an important role for ERa signaling in mesenchymal stem cell fate decisions. In addition, we found changes in lipid droplet formation, and gene expression in differentiated ERaKO adipocytes. Among others, we have identified adipocyte triglyceride lipase as a target of ERa in adipocytes. Taken together, our analysis provides further insight into the role of ERa signaling in bone marrow mesenchymal cell fate decisions and in the pathogenesis of osteoporosis.[br][br]Nothing to Disclose: KW, PAW, SAK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2299 196 1566 SUN-549 PO26-02 Sunday 1363 2012


1360 ENDO12L_SUN-550 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Differential Expression of Estrogen Receptor Subtypes in T Cell Samples from Women with Systemic Lupus Erythematosus (SLE) Virginia C Rider, Guannan Xiao, Nabih I Abdou, Bruce F Kimler Pittsburgh State University, Pittsburgh, KS; St Luke[apos]s Hospital, Kansas City, MO; Kansas University Medical Center, Kansas City, KS SLE is a strongly gender biased autoimmune disease occurring ten times more frequently in women compared with men. A fundamental problem in SLE appears to be signaling defects in peripheral T cells. The female sex hormone, estradiol 17-[beta], alters in SLE T cells signal transduction pathways that are not estradiol-responsive in normal T cells. Estradiol binds to distinct receptors, estrogen receptor alpha (ER[alpha]) and estrogen receptor beta (ER[beta]), and elicits tissue-specific responses through genomic and non-genomic mechanisms. Previous studies suggest the amount of ER[alpha] is lower in SLE T cell samples compared with healthy control T cell samples. The purpose of this study was to compare ER subtype mRNA and protein in T cells from healthy women and female patients with SLE. Blood samples were collected from healthy females (n = 10, median age 42)and SLE patients (n = 9, median age 44). The leukocytes were separated by density gradient and T cells were purified by negative selection. Total RNA and proteins were sequentially separated from the same T cell samples by column purification. ER subtype mRNA expression was measured from identical templates using real-time polymerase chain (PCR) amplification. ER[alpha] and ER[beta] proteins were quantified in the same extracts by Western blotting. Plasma estradiol was measured by ELISA. The amount of ER[alpha] protein was lower (p = 0.011, Mann-Whitney) in SLE T cell samples compared to normal T cell samples. Comparison of the ratio of protein to mRNA revealed ratios [gt] 1 for ER[alpha] while ratios for ER[beta] were generally [lt] 1. The productivity (protein: mRNA ratio) of ER[alpha] was always greater than that for ER[beta]. The comparison of relative productivity (ER[alpha]: ER[beta]) between cohorts revealed lower values in the SLE T cell samples compared with the control T cell samples (p = 0.018, Mann Whitney). Mean plasma estradiol was similar among SLE patients (110 pg/ml +/- 8.9 SEM) and healthy females (122.7 pg/ml +/- 7.6 SEM). The primary factor associated with ER[alpha] protein was ER[beta] protein (p = 0.006) and secondarily was cohort (p = 0.018). Together, the results suggest that the normal relationship between ER[alpha] and ER[beta] proteins is lost in SLE T cells. These findings are the first to identify differences in ER subtype expression in SLE T cells and point to a potential molecular mechanism underlying aberrant estradiol-dependent T cell gene regulation in this gender biased autoimmune disease.[br][br]Nothing to Disclose: VCR, GX, NIA, BFK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 278 196 1567 SUN-550 PO26-02 Sunday 1364 2012


1361 ENDO12L_SUN-551 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Extranuclear Expression of Estrogen Receptors in PC-3 Prostate Cancer Cells Raisa Pisolato, Thais FG Lucas, Maria de Fatima M Lazari, Catarina S Porto Escola Paulista de Medicina - UNIFESP, S[atilde]o Paulo, Brazil Aim: Androgen deprivation is initially effective for treatment of prostate cancer, but it may lead to an androgen-independent prostate cancer (also known as castration resistant prostate cancer). The role of 17[beta]-estradiol (E2), the classical estrogen receptors (ESR1 and ESR2), and/or the G protein[ndash]coupled estrogen receptor (GPER) in this process is unknown. ESR1 and ESR2 are predominantly localized in the nucleus of non-stimulated normal cells, but may translocate to the plasma membrane after E2 stimulation. ESRs and GPER are involved in the rapid signaling of E2, and modulate transcriptional events (reviewed in Spermatogenesis 1:318, 2011). GPER is expressed in the androgen-independent prostate cancer cell line PC-3 (Cell Death Differ.17:1511, 2010), but the expression of the classical estrogen receptors ESR1 and ESR2 in this cell line is controversial (Cancer Res. 60:3175, 2000; Eur. Urol. 40:557, 2001; Prostate 55:180, 2003). The aim of this study was to characterize the expression and cellular localization of ESR1, ESR2 and GPER in PC-3 cells. Methods and Results: PC-3 cells were grown in RPMI 1640 medium without phenol red (GIBCO), supplemented with 10% of fetal bovine serum, HEPES (5.95 mg/ml) and gentamicin (0.02 mg/ml), at 37[ordm]C, in a humidified atmosphere with 5% CO2, for 48 hours. After replacement of the medium by another, with or without serum, cells were grown for 24 and 48 hours. Western blot and immunofluorescence assays were performed as previously described (Biol. Reprod. 78:101, 2008; Biol. Reprod. 83:307, 2010). Negative controls were obtained after pre adsorption of the primary antibody with the respective blocking peptide. Specific bands of 66 kDa (ESR1), 56 kDa (ESR2) and 55 kDa (GPER) were detected in PC-3 cells. ESR1, ESR2 and GPER immunostaining was mostly found in the extranuclear region both in the absence or presence of serum. Conclusion: The results indicate that ESR1, ESR2 and GPER are possible targets for the estrogen action in the androgen-independent cell line PC-3. The preferential localization of ESR1 and ESR2 in the cytoplasm and plasma membrane region even in the absence of serum differs from the preferential nuclear localization of these receptors in normal non-stimulated cells, and may suggest that rapid signaling pathways may be constitutively activated and modulate nuclear transcriptional events. Whether these events are involved in androgen-independent prostate tumor growth still needs to be determined.[br][br]Sources of Research Support: FAPESP and CNPq.[br][br]Nothing to Disclose: RP, TFGL, MdFML, CSP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 920 196 1568 SUN-551 PO26-02 Sunday 1365 2012


1362 ENDO12L_SUN-552 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Therapeutic Significance of Estrogen Receptor [beta] Agonists in Gliomas Gangadhara R Sareddy, Binoj C Nair, Vijaya K Gonugunta, Quan-Guang Zhang, Andrew Brenner, Darrell W Brann, Rajeshwar Rao Tekmal, Ratna K Vadlamudi UT Health Sciences Center, San Antonio, TX; Georgia Health Sciences University, Augusta, GA; UT Health Sciences Center, San Antonio, TX Gliomas are the most common and devastating central nervous system neoplasms. A gender bias exists in their development: females are at lower risk than males, implicating estrogen-mediated protective effects. Estrogen functions are mediated by two ER subtypes: ER[alpha], that functions as tumor promoter and ER[beta] that function as tumor suppressor. We examined the potential use of ER[beta] agonists as a novel therapeutic to curb the growth of gliomas. Western analysis of six glioma model cells as well as primary glioma model cells showed detectable expression of ER[beta] with little or no ER[alpha]. We demonstrated that treatment of glioma cells with three distinct ER[beta] agonists results in significant decreases in proliferation. Tumor tissue microarray and IHC analysis revealed that ERb expression is down regulated in high-grade gliomas. We found that ERb agonists promote both expression and tumor suppressive functions of ER[beta] in glioma models. Liquiritigenin, a plant-derived ERb agonist significantly reduced [italic]in vivo[/italic] tumor growth in a xenograft model. Compared to control mice, animals treated with liquiritigenin had 50% reduction in tumor volume and size. IHC analysis of tumors revealed a significant increase in the nuclear ER[beta] expression with a concomitant decrease in cell proliferation in the liquiritigenin-treated group. Our results suggest that ER[beta] signaling has a tumor suppressive function in gliomas. Since ERb agonists are currently in clinical trials and are well tolerated with fewer side effects, identification of an ERb agonist as a therapeutic agent can be readily extended to clinical use with current chemotherapies, providing an additional tool for enhancing survival in glioma patients.[br][br]Nothing to Disclose: GRS, BCN, VKG, Q-GZ, AB, DWB, RRT, RKV 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1972 196 1569 SUN-552 PO26-02 Sunday 1366 2012


1363 ENDO12L_SUN-553 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Estrogen Receptor [beta] Expression Induces Changes in the microRNA Pool in Human Colon Cancer Cells Karin Edvardsson, Trang H Nguyen-Vu, Sharanya M Kalasekar, Fredrik Ponten, Jan-Ake Gustafsson, Cecilia Williams University of Houston, Houston, TX; Karolinska Institutet, Stockholm, Sweden; Uppsala University, Uppsala, Sweden A truly preventive or targeted therapy against colon cancer does not yet exist. However, hormone replacement therapy in women unexpectedly resulted in reduced risk of colorectal cancer, use of oral contraceptives is associated with a lower incidence of this disease, and estradiol has been shown to reduce the formation of preneoplastic lesions in the colon. Estrogen receptor beta (ERb) is the predominant estrogen receptor in the human colonic epithelium, and its loss is related to advanced Dukes staging. In vivo mouse studies have demonstrated that ERb agonist treatment prevents intestinal tumor development and that deletion of ERb leads to an increase in colon adenomas. Collectively, these observations clearly specify a protective role of ERb in colorectal cancer. Estrogen receptor signaling has immense consequences in health and diseases. They can be activated or inactivated by ligands, and are ideal candidates for therapeutic targeting. Compounds exist that activate only ERb, circumventing the adverse effects that estrogen induces in men and women through ERa activation. As ERb hold significant potential as a target for colon cancer therapy, it is essential to understand the basic mechanistic background of its action and to identify biomarkers of its activity. We have previously shown that ERb is antiproliferative, has antitumorigenic properties and that its expression induces transcriptome wide changes in colon cancer cells. It is plausible that ERb also regulates microRNAs (miRNAs) and that these contribute to the ERb mediated protective effects. Here, we investigate whether or not ERb induces a change in the miRNA pool in colon cancer cells and if such changes contribute to the ERb mediated protective effects.[br]We detected and confirmed 28 miRNAs to be changed following ERb expression in SW480 colon cancer cells. The majority of the changed miRNAs were being downregulated. These include miR-200a and b, miR-9 and the entire miR-17-92 cluster. Three miRNAs were significantly upregulated; miR-10a and b, and miR-205. We explored their regulation and effect on a genome-wide scale and describe functional studies of the most regulated and implicated miRNAs: miR-17 and miR200a and b. We showed that miR-17 regulates CLU and NCOA3/SRC-3 thereby affecting cisplatin treatment. Further, we showed that miR-200a and b regulate MYC, ZEB1 and E-cadherin and that miR-200a affects proliferation.[br][br]Nothing to Disclose: KE, THN-V, SMK, FP, J-AG, CW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1971 196 1570 SUN-553 PO26-02 Sunday 1367 2012


1364 ENDO12L_SUN-554 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Exploring Diverging Functions of Long-Term ER[beta] Expression in Two Breast Cancer Cell Lines Philip Jonsson, Anders Strom, Cecilia Williams University of Houston, Houston, TX Breast cancer is the second-most common cancer among women in the US. Tumors are typically classified by the molecular subtype whereof one is the estrogen-receptor positive tumors. Disruption of estrogen signaling is used in the clinic by targeting ER[alpha] with antagonists, e.g. tamoxifen. Animal models and cell lines support this notion, contributing to understanding the underlying mechanisms. For the second estrogen receptor, ER[beta], in vitro studies suggest a protective role in tumorigenesis. However, recent studies indicate that 40 to 80 percent of tumors express both ERs.[br]To model ER[alpha]+/[beta]+ tumors we are studying ER[alpha]+ breast cancer cell lines MCF7 and T47D, transduced by lentivirus to express ER[beta]. The transcriptome changes upon expressing ER[beta] differ vastly in the two cells lines, suggesting cell-line specific functionality. Remarkably, pro-proliferation factors such as MYC, CCNA2 and PCNA are increased in MCF7 but not T47D when ER[beta] is expressed. Similarly, the estrogen response after ER[beta] introduction varies between the cell lines; addition of estrogen enhances the ER[alpha]-dependent response on some genes but counteracts it on others. Especially in MCF7 the ER[alpha] response is enhanced, also at the basal expression level of many genes. In accordance with observed gene-expression profiles, ER[beta] expression in MCF7 increases proliferation even in absence of ligand, whereas no change in proliferation occurs in T47D. The increased proliferation of MCF7-ER[beta] is ablated by ICI and the estradiol-induction is suppressed by tamoxifen. Interestingly, the expression of the transcription factor GATA3, associated with favorable breast-cancer prognosis and proposed to play a role in ER signaling, is decreased by ER[beta] in MCF7 and increased in T47D. Additionally, we demonstrate the presence of ER heterodimers in MCF7.[br]In conclusion, the expression of ER[beta] confers transcriptome changes that vary both cell lines and across genes, suggesting that ER[beta] not always acts to oppose ER[alpha]. Our data contradict most previous in vitro data on ER[beta][apos]s function in breast cancer cells but is concordant with previously mentioned studies that found tumors positive for ER[beta]. Most previous studies have been performed using transient expression of ER[beta], whereas this is one of few that study the effect of constitutive expression. It is possible that, depending on the cellular context, cancer cells can proliferate in the presence of ER[beta] and in some cases even thrive in it, as is the case for our MCF7 cells.[br][br]Nothing to Disclose: PJ, AS, CW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1994 196 1571 SUN-554 PO26-02 Sunday 1368 2012


1365 ENDO12L_SUN-555 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Regulation of microRNA by Estrogen-Activated Estrogen Receptor [beta] in Breast Cancer Anne Chinenye Katchy, Cecilia Williams University of Houston, Houston, TX Estrogens (E2) play a role in breast cancer progression by regulating genes involved in certain cellular functions through the estrogen receptors (ER). MicroRNAs are short, non-coding RNAs that regulate genes by either blocking translation or by mRNA degradation. Thus, identifying miRNAs that are associated with breast cancer progression by normal or disrupted estrogen signaling would provide new insights to the prognosis and diagnosis of breast cancer. We have previously shown that no significant regulation of miRNAs by 24 hours E2-activated ER[alpha] (E2-ER[alpha]) in T47D breast cancer cell line. Noting both ER receptors have antagonizing functional relationship with each other and the fact that less is known about ER[beta] regulation of miRNAs, we proceeded to investigate if 24 hours E2-ER[beta] regulates miRNAs in this cell line. To identify miRNAs regulated by E2-ER[beta], miRNAs expression profiles were analyzed using dual-color microarrays and validations were carried out using real-time PCR. Several miRNAs were found to be regulated by E2-ER[beta] such as miR-22, miR-522, and let7b, which are in turn, involved in several cellular functions, such as miR-22 which is a known ER[alpha] suppressor and inhibits estrogen signaling by targeting ER[alpha] mRNA. A thorough investigation of ER regulation of miRNAs would bring us closer to understand the mechanism of action of the estrogen receptors.[br][br]Nothing to Disclose: ACK, CW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1836 196 1572 SUN-555 PO26-02 Sunday 1369 2012


1366 ENDO12L_SUN-556 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Estrogen Receptor Beta Splice Variants in Malignant Pleural Mesothelioma: Prognostic Indicators and Therapeutic Targets Cormac J Jennings, Anthony O[apos]Grady, Robert Cummins, Bruno Murer, Elaine W Kay, Brian J Harvey, Warren Thomas Royal College of Surgeons in Ireland, Dublin, Ireland; Royal College of Surgeons in Ireland, Dublin, Ireland; Dell[apos] Angelo Hospital, Mestre, Italy [bold][underline]Introduction[br][/underline][/bold]Malignant Pleural Mesothelioma (MPM) is a highly aggressive tumor with poor prognosis that arises from the mesothelial lining of the lungs. Asbestos exposure is the greatest risk factor in the development of MPM; however, the malignancy is generally diagnosed decades after initial exposure. The incidence of MPM is expected to peak in the majority of developed nations over the next two decades (1). Female gender is a positive prognostic indicator for MM progression and post diagnosis survival (2, 3). This study investigated the expression of progesterone receptor (PR), estrogen receptors alpha (ER[alpha]), beta 1 (ER[beta]1) and beta 2 (ER[beta]2) in normal mesothelial and MPM tumor samples and relationship with clinical outcome. The contribution made by these receptors to modulating MPM cell growth was also investigated.[br][bold][underline]Methods[br][/underline][/bold]Immunohistochemistry (IHC) analysis was performed to determine the expression of PR, ER[alpha], ER[beta]1 and ER[beta]2 in tumor biopsies from a cohort of 89 confirmed MPM subjects and 3 normal mesothelial controls. Allred scores were assigned based on the intensity and percentage expression of each of the proteins, and Kaplan-Meier survival curves were generated. Real time (RT)-PCR was performed to compare receptor expression profiles of 11 MPM cell lines. MTS assays were used to assess the effect of ER isoform expression on cell growth.[br][bold][underline]Results[/underline][/bold][br]High levels of ER[beta]1 and ER[beta]2 expression were found in normal mesothelial tissue samples, while variable levels of expression were found in MPM tumor specimens. ER[alpha] or PR was not detected by IHC in normal mesothelial or in MPM tissue tumor sections. Median survival for ER[beta]1 positive patients was 15 months compared to 8 months in ER[beta]1 negative patients (p = 0.254, log-rank test), while median survival for ER[beta]2 positive patients was 14 months compared to 7 months in ER[beta]2 negative patients (p = 0.036, log-rank test). RT-PCR results indicated that ER[beta]1 and ER[beta]2 mRNA levels varied between MPM cell lines, while ER[alpha] and PR were not expressed. Heterologous over-expression of ER[beta]1 in the REN MPM cell line caused a significant reduction in cell viability after 72 hours, both in the presence (30% [plusmn] 5 reduction) and absence (20% [plusmn] 5 reduction) of estrogen.[br][bold][underline]Conclusions[/underline][/bold][br]Modulation of ER-coupled signalling may provide novel approaches to therapeutic intervention for this terminal malignancy. The improved outcome for patients that express ER[beta] isoforms may provide novel steroid-based approaches to therapeutic intervention for MM.[br][br]1. Lee et al., Cancer 2007; 109 : 1454-61. 2. Flores et al., J. Thor. Oncol. 2007; 2:957-965. 3. Jennings et al., J. Thor. Oncol. 2012; 7:243-248.[br][br]Sources of Research Support: This work was funded by the Health Research Board in Ireland under Grant No. PHD/2007/11.[br][br]Nothing to Disclose: CJJ, AO, RC, BM, EWK, BJH, WT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2175 196 1573 SUN-556 PO26-02 Sunday 1370 2012


1367 ENDO12L_SUN-557 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Role of Progesterone Receptor (PR) Sumoylation Motif and DNA-Binding Domain in Progesterone (P4) Repression of NF-[kappa]B Transcriptional Activity Chien-Cheng Chen, Carole R Mendelson University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX Progesterone/PR plays a crucial role in maintaining myometrial quiescence throughout pregnancy, while labor is associated with increased expression of proinflammatory cytokines (e.g. IL-1[beta]) and activation of nuclear factor-[kappa]B (NF-[kappa]B), resulting in increased expression of genes promoting myometrial contractility. Previously, we observed that P4/PR plays an anti-inflammatory role by antagonizing stimulatory effects of IL-1[beta]on cyclooxygenase 2 (COX-2) expression in human myometrial cells. These actions of P4 occur both by direct PR interaction with NF-[kappa]B and by induction of I[kappa]B[alpha], resulting in decreased binding of NF-[kappa]B p65 to the COX-2 promoter. Sumoylation of PR at K388 has been reported to enhance its capacity to mediate transcriptional repression, while the PR DNA-binding domain was found to interact with NF-[kappa]B p65. Thus, to further study mechanisms for this repression, HEK 293 cells stably expressing PR-B (WT), a sumoylation site mutant PR-B (K388R) or a DNA-binding domain mutant PR-B (mDBD) were generated. When cells were transiently transfected with a luciferase reporter plasmid containing NF-[kappa]B consensus binding sites, treatment with IL-1[beta] caused a 5.5-fold induction of luciferase activity compared to vehicle, while treatment with IL-1[beta] + P4 caused a [sim]50% decrease in luciferase activity compared to IL-1[beta] alone. However, in cells stably expressing PR-B (K388R) or PR-B (mDBD), P4 lost its ability to repress IL-1[beta]-induced luciferase activity. When cells expressing PR-B (WT) were transfected with luciferase reporter plasmids under control of the native COX-2 or IL-6 promoters, treatment with IL-1[beta] + P4 significantly inhibited luciferase activity compared to IL-1[beta] alone, while in cells stably expressing PR-B (K388R) or PR-B (mDBD), P4/PR repressive activity was partially lost. On the other hand, when RT-qPCR was used to analyze effects of IL-1[beta] and P4 on endogenous COX-2 and IL-6 expression, both PR-B (WT) and the PR-B sumoylation mutant (K388R) were able to mediate P4 repression of IL-1[beta] stimulation, while in cells stably expressing the PR-B (mDBD) mutant, P4 repression of IL-1[beta] induction of COX-2 and IL-6 expression was lost. Taken together, these findings suggest that both the sumoylation site and DBD of PR may play a role in repression of NF-[kappa]B mediated transcription; however, their relative importance appears to differ according to the reporter platform chosen.[br][br]Sources of Research Support: NIH-R01-DK31206 and NIH-P01-HD-11149.[br][br]Nothing to Disclose: C-CC, CRM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2362 196 1574 SUN-557 PO26-02 Sunday 1371 2012


1368 ENDO12L_SUN-558 POSTER SESSION: Estrogen [amp] Progesterone Receptors in Normal Development [amp] Disease (1:30 PM-3:30 PM) Stat5 as a Coregulator of Progesterone Receptor Target Genes in Normal Mammary Epithelial Cells Alison E Obr, Sandra L Grimm, John P Lydon, Kathleen A Bishop, J Wesley Pike, Dean P Edwards Baylor College of Medicine, Houston, TX; University of Wisconsin, Madison, WI Progesterone (P4), a proliferative hormone in the mammary gland at different stages of development, is required for ductal side branching during puberty and for epithelial expansion and alveologenesis during pregnancy. Previous gene microarray analysis of adult virgin mice treated acutely with P4 identified subsets of cell cycle and paracrine growth factor genes as PR targets and potential mediators of P4 induced cell proliferation (1). To study mechanisms of PR regulation of these target genes, we have developed a 3D culture system of primary mammary epithelial cells (MECs) embedded in Matrigel that form polarized acini composed of luminal and myoepithelial cells and maintain a heterogeneous pattern of PR expression in non-dividing luminal cells similar to that in the mammary gland in vivo. As assessed by high throughput TaqMan Array qPCR, P4 induced expression of most of the same target genes and in a similar manner as in the mammary gland in vivo, including paracrine factors RANKL, Wnt-4 and amphiregulin, and the pro-proliferative transcription factors, Gata3 and Id4. RANKL has emerged from recent in vivo studies to be an essential paracrine effector of P4 proliferation in the mammary gland, thus we have focused on RANKL for mechanistic studies in vitro. In co-transfection experiments with a mouse cell line, NMuMg, P4 in a PR dependent manner induced activation of 3 different RANKL enhancers linked to a luciferase reporter and as detected by ChIP assay PR bound to these same RANKL enhancers in response to P4. By ChIP assay Stat5 was also recruited in a P4 dependent manner to the same enhancer regions as PR in the absence of prolactin (PRL) that is required to activate Stat5 as a sequence specific transcriptional activator. To determine whether Stat5 has a role in PR-mediated gene regulation, primary MEC 3D cultures derived from Stat5 knockout mice (provided by Dr. Yi Li, BCM) were analyzed. P4 induction of RANKL, Wnt-4, amphiregulin and Gata3 was substantially reduced in MECs from Stat5 KO mice as compared with wildtype MECs. These data indicate that RANKL is a direct PR target gene and that Stat5 is a co-activator on RANKL and other PR target genes. To further elucidate the role of Stat5 in P4/PR transcriptional signaling in the mammary gland, genome-wide binding of PR and Stat5 by ChIP-chip analysis of primary MECs is currently being conducted and will be reported.[br][br]Fernandez-Valdivia, R et al., Endocrinology 2008; 149:6236.[br][br]Nothing to Disclose: AEO, SLG, JPL, KAB, JWP, DPE 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1958 196 1575 SUN-558 PO26-02 Sunday 1372 2012


1369 ENDO12L_SUN-559 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) The Androgen Metabolite, 5[alpha]-Androstane-3[beta],17[beta]-Diol, Decreases Cytokine-Induced Cyclooxygenase-2 and Vascular Cell Adhesion Molecule-1 Expression, and Reduces P-Glycoprotein Expression in Human Brain Microvascular Endothelial Cells Kristen L Zuloaga, Sibyl N Swift, Rayna J Gonzales, T John Wu, Robert J Handa University of Arizona College of Medicine - Phoenix, Phoenix, AZ; Oregon Health [amp] Science University, Portland, OR; Uniformed Services University of the Health Sciences, Bethesda, MD P-glycoprotein (Pgp), a multiple drug resistance transporter expressed by vascular endothelial cells, is a key component of the blood-brain barrier (BBB) and has been shown to increase following inflammation. The non-aromatizable androgen, dihydrotestosterone (DHT), decreases inflammatory markers in vascular smooth muscle cells in an androgen receptor (AR) independent fashion. The principal metabolite of DHT, 5[alpha]-androstane-3[beta],17[beta]-diol (3[beta]-diol), binds estrogen receptor (ER) [beta] and also decreases inflammatory markers in endothelial cells. Therefore, we tested the hypothesis that DHT and 3[beta]-diol decrease expression of IL-1[beta]-induced pro-inflammatory mediators, vascular cell adhesion molecule-1 (VCAM-1) and cyclooxygenase-2 (COX-2), to regulate Pgp expression in primary human brain microvascular endothelial cells (HBMEC). Messenger-RNAs for AR, ER[alpha], and ER[beta] and the steroid metabolizing enzymes necessary for DHT conversion to 3[beta]-diol were detected in HBMEC cells demonstrating that the cellular machinery for production of and responsiveness to 3[beta]-diol are present. Subsequently, Western blot analysis of HBMEC lysates showed that 3[beta]-diol (10nM) reduced COX-2 and Pgp expression and the effect of 3[beta]-diol on Pgp was prevented by the ER antagonist, ICI-182,780 (ICI, 1[mu]M). Treatment with the cytokine, IL-1[beta],(5ng/ml) caused increases in COX-2 and VCAM-1 and these effects were reduced by co-treatment with either DHT (10nM) or 3[beta]-diol (10nM). 3[beta]-diol (10nM) also decreased cytokine-induced Pgp expression. ICI (1[mu]M) blocked the effect of 3[beta]-diol (10nM) on COX-2 and VCAM-1, but not on Pgp expression. Therefore, in cytokine-stimulated HBMEC, the effect of 3[beta]-diol on cerebrovascular pro-inflammatory mediator expression is ER dependent, whereas its effect on Pgp expression is ER independent. These studies suggest a novel role of 3[beta]-diol in regulating BBB function and support the concept that 3[beta]-diol can reduce cerebral vascular inflammation.[br][br]Sources of Research Support: NIH NS039951 and DoD: DMRDP# D61_I_10_J6_125.[br][br]Nothing to Disclose: KLZ, SNS, RJG, TJW, RJH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1995 197 1576 SUN-559 PO24-02 Sunday 1373 2012


1370 ENDO12L_SUN-560 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Anti-Inflammatory Activity of Estrogen-ER[alpha] Axis Eugenia Morselli, Rafael Nafikov, Lisa Hahner, Deborah Clegg University of Texas Southwestern Medical Center, Dallas, TX Obesity has become a major health problem worldwide. In the USA, it affects over the 30% of the adult population. The chronic low[ndash]grade inflammatory response associated with obesity is known to contribute to different diseases such as diabetes, cardiovascular diseases and cancer. Estrogens, acting through the estrogen receptor alpha (ER[alpha]), have anti-inflammatory properties decreasing the number of pro-inflammatory cytokines [italic]in vitro [/italic]and [italic]in vivo[/italic]. Additionally, it has been appreciated that males and females differ with respect to disease prevalence, metabolism and inflammatory response. Therefore, our goal has been to evaluate whether estrogens, acting through the ER[alpha] signaling pathway, can inhibit inflammation and if this response is sexually dimorphic. We analyzed the inflammatory response of different tissues derived from male and female C57BL6 mice exposed to high fat diet (HFD) and then injected with lipopolysaccharide (LPS) or saline solution. While all tissues from males and females responded to LPS injection, the inflammatory response to HFD and ER[alpha] expression was tissue- and sex- dependent. Interestingly, we found a sexually dimorphic response in tissues from hippocampus and liver. In these tissues, HFD-induced inflammation was higher in males (IL-6, TNF[alpha], TLR4 and NALP3 expression was evaluated), where ER[alpha] expression is lower, than in females, where ER[alpha] expression is higher. Additionally, we found injections with LPS significantly reduced the expression of ER[alpha] in male tissues while in females the expression was increased. Our data support the idea in females estrogens, acting through ER[alpha], provide protection against inflammation both in the central nervous system and in the periphery. Importantly, our data further suggest the mechanism by which estrogens and ER[alpha] protect against inflammation is through induction of autophagy.[br][br]Nothing to Disclose: EM, RN, LH, DC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1967 197 1577 SUN-560 PO24-02 Sunday 1374 2012


1371 ENDO12L_SUN-561 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Estrogen Signaling in Innate Immunity Elisabetta Vegeto, Stefania Masiero, Adriana Maggi University of Milan, Milan, Italy Estrogens are pleiotropic hormones that act through the interaction with intracellular receptors, ERalpha and ERbeta, which are ligand-dependent transcription factors expressed in several tissues and cell lineages. Clinical evidence suggests that a decrease in the levels of circulating estrogens, as it occurs in women at menopause, is associated with a higher incidence of inflammation-associated pathologies, such as osteoporosis and neurodegenerative diseases; accordingly, estrogens administration correlates with a protective effect against the onset and progression of these pathologies also through the reduction of the inflammatory response. We thus hypothesized that estrogens exert beneficial effects by acting as anti-inflammatory agents. Our experiments on ERs gene expression show that resident macrophages express both ERs at different levels (1). Using experimental models of acute inflammation in brain and lung, we observed that 17beta-estradiol reduces the induction of inflammatory genes expression in vivo through the activation of ER (1-3). Lack of ERalpha results in increased plasma membrane expression of Mac-1 integrin (C3R), a critical protein in cell adhesion/motility, host defense against pathogens and, as recently observed, in neuroinflammation associated with neurodegenerative pathologies. We are thus studying ERs signaling in macrophages and evaluating their role in gene expression, motility and activation of these cells in order to have a deeper understanding of the role of estrogens in the physiology and reactivity of innate immunity. The results of these studies will be discussed.[br][br][br]1. Vegeto et al (2010) Endocrinology 15, 174-84[br]2. Vegeto et al (2003) Proc Natl Acad Sci USA 100, 9614-9[br]3. Vegeto et al (2006) Endocrinology 147, 2263-72[br][br]Nothing to Disclose: EV, SM, AM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1425 197 1578 SUN-561 PO24-02 Sunday 1375 2012


1372 ENDO12L_SUN-562 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Cooperative Activation of Gene Expression by Agonists and Antagonists Can Be Mediated by Estrogen Receptor Heteroligand Dimer Complexes Shuang Liu, Sang Jun Han, Carolyn L Smith Baylor College of Medicine, Houston, TX Estrogen receptor (ER) antagonists such as tamoxifen inhibit estrogen action through competitive inhibition resulting in receptor binding to antagonist rather than agonist. However, microarray analyses reveal a group of genes for which ER agonist and antagonist cooperatively regulate expression, suggesting additional models of combined agonist/antagonist action must exist. It was hypothesized that ER heterodimers consisting of one receptor monomer bound to agonist and another occupied by antagonist could mediate the stimulatory effects of combined agonist and antagonist. To test whether a ER heteroligand dimer (ER-HLD) complex could cooperatively regulate gene expression, a chimeric receptor and reporter system was generated consisting of: 1) ER[alpha]-G521R possessing a ligand binding domain mutation permitting antagonist but not estradiol binding; 2) ER[alpha](GSCKV) encompassing a DNA binding domain mutation enabling binding to a glucocorticoid response element (GRE) and 3) chimeric reporter genes in which a luciferase cDNA is downstream of a composite, half-site GRE/half-site estrogen response element (ERE). In either HeLa or HepG2 cells transfected with expression vectors for ER[alpha]-G521R and ER[alpha](GSCKV), and a chimeric reporter, combined estrogen and antagonist treatment yielded greater luciferase expression than for either agent alone. Neither receptor alone was able to recapitulate this response. Co-immunoprecipitation experiments of Flag-ER[alpha]-G521R and HA-ER[alpha](GSCKV) revealed a basal level of receptor interaction that could be stimulated by combined agonist/antagonist treatment. The absence of a synergistic effect on a half-site ERE reporter indicates that agonist-bound ER[alpha](GSCKV) and antagonist-bound ER[alpha]-G521R must both bind to DNA to cooperatively activate gene expression. Ligand combinations able to induce gene expression in this model system include the agonists 17[beta]-estradiol or conjugated estrogens with the antagonists tamoxifen, raloxifene, bazedoxifene or fulvestrant. Moreover, agonist-bound ER[alpha] and antagonist-bound ER[alpha]-G521R cooperatively enhance the activity of a complement 3-luciferase reporter demonstrating that ER-HLDs can activate transcription in the context of a natural promoter. Overall, this finding broadens our understanding of the complex relationship between estrogens and antiestrogens, and suggests a novel mechanism by which cell and tissue selective effects of TSECs (tissue selective estrogen complexes) may be achieved.[br][br]Disclosures: CLS: Advisory Group Member, Pfizer, Inc.; Investigator, Pfizer, Inc. Nothing to Disclose: SL, SJH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1945 197 1579 SUN-562 PO24-02 Sunday 1376 2012


1373 ENDO12L_SUN-563 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Comprehensive Profiling of the Estrogen-Regulated Transcriptome in Native Mouse Tissues Christina K Matulis, Anusha Nagari, William Lee Kraus University of Texas Southwestern Medical Center, Dallas, TX In mammals, the steroid hormone estrogen exerts gene regulatory effects during many physiological processes, including those involved in mammalian development and reproduction, metabolism and lipogenesis, cardiovascular and neurological processes, and carcinogenesis. Estrogens, acting through estrogen receptors (ER[alpha] or ER[beta]), regulate gene expression in a tissue and cell-type specific manner to control physiological outcomes. Many previous studies have used genomic approaches to identify the ER cistrome (i.e., the location of ER binding sites), as well as the steady-state levels of protein-coding RNAs whose expression is regulated by estrogen signaling, in a variety of model systems (e.g., cultured cells, mouse tissues). However, our view of the estrogen-dependent gene regulatory effects that occur as a result of primary transcriptional responses is limited. In addition, how estrogen signaling affects the expression of genes for non-coding RNAs (ncRNAs), such as long non-coding RNAs (lncRNAs), and the role of these ncRNAs in the molecular mechanisms that control estrogen-regulated gene transcription, have not been clearly elucidated. We are using Global Run-on Sequencing (GRO-seq), a novel genomic approach that combines high throughput sequencing and computational algorithms, to map the location, orientation, and abundance of all active RNA polymerases (Pol I, Pol II, and Pol III) across the genome in response to estrogen signaling. GRO-seq also allows the identification of previously unannotated transcription units. Previously, we used GRO-seq to define a set of coding and non-coding RNAs that are rapidly and transiently regulated by estrogen in MCF-7 human breast cancer cells. To elucidate the molecular mechanisms by which estrogens exert their gene regulatory effects in different biological systems in vivo, we have performed GRO-seq using tissues (i.e., mammary gland, uterus, liver) collected from estrogen treated mice. Analysis of these data sets is allowing us to map the immediate transcriptional effects of estrogens in vivo, as well as characterize novel ncRNAs that may function as key components of the estrogen signaling pathway. A better understanding of the direct estrogen-regulated transcriptome will provide new insights into estrogen signaling and may aid in finding better ways to prevent, diagnose, and treat estrogen-related diseases.[br][br]Sources of Research Support: This work is supported by a grant from the NIH/NIDDK to W.L.K.[br][br]Nothing to Disclose: CKM, AN, WLK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1003 197 1580 SUN-563 PO24-02 Sunday 1377 2012


1374 ENDO12L_SUN-564 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Role of Estrogen Receptor-Dependent Enhancer RNAs in Estrogen-Regulated Transcriptional Responses Shino Murakami, Nasun Hah, Anusha Nagari, Charles G Danko, W Lee Kraus University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX; The Salk Institute for Biological Studies, La Jolla, CA; Cornell University, Ithaca, NY; Cornell University, Ithaca, NY Estrogen signaling controls a wide array of important physiological and pathological responses. Many of the effects are mediated by the direct or indirect binding of estrogen receptors (ERs) to sites across the genome, which ultimately leads to regulated transcriptional responses at nearby or distally-located target genes. Emerging evidence has indicated that the binding of transcription factors, such as ERs, to genomic DNA results in the bi-directional synthesis of short, non-coding RNAs in the vicinity of the binding sites. The function of these enhancer RNAs (eRNAs) is not well understood. We have used Global Run-On and sequencing (GRO-seq), in combination with a set of a novel bioinformatic approaches, to map and characterize the eRNAs arising from ER binding sites in ER-positive MCF-7 human breast cancer cells in response to short treatments with estradiol. Using this approach, we identified more than 700 eRNAs originating at distal ER binding sites (i.e., [gt]10 kb from the TSS) in MCF-7 cells. Two thirds of the eRNAs are paired with a divergent transcript generated on the opposite strand, while the remainder are transcribed without a divergent transcript. The expression of 40% of the total eRNA set was estrogen-regulated, with the majority being up-regulated. We are now examining the mechanisms by which these transcripts are regulated, their effects on ER binding and chromatin-dependent gene loops, and the regulation of neighboring estrogen-regulated target genes. In addition, we are using these analyses to identify and map novel active enhancers in MCF-7 cells. These studies are revealing more complexity in the ER cistrome and the estrogen-regulated transcriptome than was previously known.[br][br]Sources of Research Support: This work is supported by a grant from the NIH/NIDDK to W.L.K. and a postdoctoral fellowship from the PhRMA Foundation to C.G.D.[br][br]Nothing to Disclose: SM, NH, AN, CGD, WLK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2014 197 1581 SUN-564 PO24-02 Sunday 1378 2012


1375 ENDO12L_SUN-565 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Identification and Functional Characterization of Estrogen-Regulated Long Noncoding RNAs in Breast Cancer Cells: From Genomics to Molecular and Cellular Biology Shrikanth S Gadad, Miao Sun, W Lee Kraus University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX; Cornell University, Ithaca, NY Estrogen signaling controls a wide array of important physiological and pathological responses, many of which occur as a result of direct transcriptional effects of estrogen on the genome, mediated by estrogen receptors. Most previous studies examining the transcriptional effects of estrogen have focused on protein coding gene. Nevertheless, emerging evidence indicates that estrogen signaling also affects the expression of many genes whose products are non-coding functional RNA transcripts, including long non-coding RNAs (lncRNAs). LncRNAs are a poorly characterized and annotated class of polyadenylated and spliced RNAs that are transcribed from genic or intergenic regions of the genome. They do not code for proteins; rather, they function directly as RNAs and have been implicated in transcriptional regulation and epigenetic control. Previously work in our lab used Global Run-On and sequencing (GRO-seq) to examine the immediate effects of estrogen signaling on the transcriptome of MCF-7 human breast cancer cells. We have developed a novel bioinformatics approach to call primary transcripts of all non-coding RNAs from the GRO-seq data. We have combined this transcript calling with RNA-seq and additional bioinformatic analyses to define a set of over 1400 expressed lncRNAs in MCF-7 cells, about two-thirds of which have not been annotated previously. These lncRNAs have little coding potential, and about a quarter of them are estrogen-regulated. In functional assays, knockdown of selected estrogen-regulated lncRNAs alters the expression of neighboring protein-coding genes and inhibits MCF-7 cell growth. We are now performing detailed molecular analyses to better understand the underlying mechanisms of action of the lncRNAs. These studies are providing a much greater understanding of estrogen-dependent gene regulation than has been achieved with mRNA profiling alone. In addition, the lncRNA transcriptome is likely to have a high level of utility as a diagnostic and prognostic tool in a variety of diseases, including cancers, and it may also suggest new targets for therapeutic interventions.[br][br]Sources of Research Support: This work is supported by a grant from the NIH /NIDDK to W.L.K.[br][br]Nothing to Disclose: SSG, MS, WLK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 612 197 1582 SUN-565 PO24-02 Sunday 1379 2012


1376 ENDO12L_SUN-566 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Regulation and Function of Antisense Transcription in the Estrogen Response in Breast Cancer Cells Rui Li, Minho Chae, Miao Sun, W Lee Kraus University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX; Cornell University, Ithaca, NY Estrogen signaling plays key roles in a wide array of physiological processes and disease states. Many of these effects are through the gene regulatory actions of estrogens, which act through estrogen receptor proteins to induce genome-wide alterations in the expression of protein-coding and non-coding genes. We are using a variety of gene-specific and genomic approaches in MCF-7 human breast cancer cells to examine the effects of estrogen signaling on the production of antisense transcripts, which overlap with and are transcribed in the opposite direction from protein-coding genes. We have used Global Run-On and sequencing (GRO-seq), a genomic approach that maps the location and direction of all active RNA polymerases, combined with de novo transcript calling, to identify about [sim]1,200 antisense transcription units. We are now using RNA-seq to map the steady-state RNAs that are produced from these transcription units. Little is known about antisense transcription in mammalian cells and the mechanisms by which it may affect sense gene expression or biological processes are poorly understood. Is antisense transcription required for transcription of the overlapping sense gene? Do antisense transcripts target and regulate the sense transcripts? We are now addressing these fundamental questions. Our results indicate that sense and antisense transcription are largely co-regulated upon estrogen treatment [i.e., similar direction (up or down) and magnitude of regulation]. Strand-specific RT-qPCR analyses have allowed us to detect the antisense RNAs and monitor their expression in response to estrogen treatment. We are characterizing these antisense RNAs by cloning and RNAi-mediated knockdown. We are also examining the mechanisms that regulate antisense transcription, as well as their potential biological roles in human breast cancer cells. These studies will shed new light on the structure, function, and regulation of antisense transcripts.[br][br]Sources of Research Support: A grant from the NIH/NIDDK to W.L.K.[br][br]Nothing to Disclose: RL, MC, MS, WLK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2004 197 1583 SUN-566 PO24-02 Sunday 1380 2012


1377 ENDO12L_SUN-567 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) A Direct Signaling Mechanism Involved in Estradiol and Diethylstilbestrol Effects on Androgen-Dependent and Androgen-Independent Prostate Cancer Cell Viability Luke Koong, Cheryl S Watson University of Texas Medical Branch, Galveston, TX Prostate cancer typically affects men over the age of 65 and begins as androgen-dependant tumors. Treatment with anti-androgens is usually effective as therapy, but some patients relapse with androgen-independent tumors. DES, achieving [micro]M blood levels, is a common therapy for these advanced tumors and is believed to decrease androgen production in males. However, we hypothesized a more direct mode of action for estrogens and the aim of this study was to measure changes in cell viability, extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase phospho-activation, and caspase activation resulting from estradiol (E[sub]2[/sub]) or DES treatment. Using the MTT assay after three days of treatment, the viability of androgen-dependant (LAPC-4) prostate cancer cells was decreased as much as 30% by E[sub]2[/sub], and a maximum of 20% only at the lowest concentrations (10[sup]-14[/sup]-10[sup]-12[/sup] M) of DES. Both E[sub]2[/sub] and DES were slightly effective in androgen-independent (PC-3) prostate cancer cells (reducing viability 5-10%). ERK and JNK are thought to be involved in the control of cell proliferation, and we measured their rapid, non-genomic activation using our plate immunoassay. In LAPC-4 cells, E[sub]2[/sub] phospho-activated ERK after 10 min., but deactivated JNK after 5 min. DES deactivated both kinases after 5 min. In PC-3 cells, E[sub]2[/sub] activated ERK after 15 min. and JNK after 5 min. DES did not activate ERK or JNK at the time points up to 60 min. Activated caspases are involved in mediating cellular apoptosis, and we measured the activation of caspase 8 and 9 after 16 hours of estrogen treatment by the cleavage of a fluorescent substrate. E[sub]2[/sub] slightly but significantly induced caspase 8 in LAPC-4 cells, while DES activated both caspase 8 ([gt]350%) and 9 (slightly but significantly) in PC-3 cells. In summary, E[sub]2[/sub] and DES act directly on prostate cancer cells to alter cell number outcomes. MAPK activation does not appear to follow traditional signaling paradigms in LAPC-4 cells, regardless of estrogen treatment, and this supports recent findings that ERK can stimulate apoptosis via alternative signaling pathways involving radical oxygen species activation of caspases. PC-3 cells on the other hand appear to follow a more traditional model of MAPK signaling leading to cell death. A difference in membrane estrogen receptors expressed by these cells could account for this difference, and will be the focus of future studies.[br][br]Sources of Research Support: NIEHS Environmental Toxicology Training Grant T32-ES07254 awarded to LK.[br][br]Nothing to Disclose: LK, CSW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1743 197 1584 SUN-567 PO24-02 Sunday 1381 2012


1378 ENDO12L_SUN-568 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Rapid Estrogen Signaling Increases IL-8 Levels Via NF[kappa]B Activation in Endometrial Cancer Cells Melanie Scully, Twila Jackson University of Colorado Denver, Aurora, CO Each year, approximately 142,000 women worldwide develop endometrial cancer, and an estimated 42,000 women die from the disease. Estrogens (E2), acting through the estrogen receptor (ER), regulate proliferation in the endometrium, and hyper-estrogenicity is thought to be the primary cause of type 1, hormone-dependent, endometrial cancer. While ER have historically been thought of as ligand-dependent transcription factors, recent observations suggest that E2 and other steroid hormones can elicit rapid activation of cellular signal transduction pathways. However, the role of rapid, E2-activated ER signaling in endometrial cancer cell growth is not well studied.[br]The PTEN tumor suppressor regulates cell growth by antagonizing the PI3 kinase/Akt pathway. We have previously shown that rapid E2 signaling regulates PTEN activity and stability via carboxy-terminal phosphorylation. PTEN activity is decreased by C-terminal phosphorylation and stability is increased.[br]We hypothesize that decreased PTEN activity leads to hyper-stimulation of the PI3K/Akt signaling axis and subsequent activation of NF[kappa]B leads to a pro-inflammatory, pro-tumorigenic microenvironment. To test this hypothesis, endometrial cancer cells expressing ER and PTEN were treated with E2 over a short time course. We show that E2 treatment of endometrial cancer cells induces NF[kappa]B subunit RelA translocation to the nucleus within 5 minutes. Use of inhibitors shows decreased and delayed RelA nuclear translocation. Furthermore, we show a known transcriptional target of NF[kappa]B, IL-8 mRNA, is also increased.[br]We show that rapid E2/ER actions upregulate the pro-inflammatory cytokine IL-8 in endometrial cancer cells. Our data suggest that this is through down regulation of PTEN activity. Due to the high levels of circulating E2 observed in women at high risk for endometrial cancer, PTEN may be aberrantly maintained in a low activity state thus allowing for pro-inflammatory, pro-tumorigenic microenvironment. Our results suggest that misregulation of E2/ER non-genomic signaling could provide a widespread mechanism for tumorigenesis.[br][br]Sources of Research Support: NCI RO1 CA 125427.[br][br]Nothing to Disclose: MS, TJ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2323 197 1585 SUN-568 PO24-02 Sunday 1382 2012


1379 ENDO12L_SUN-569 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Estrogenic Action on Arterial Smooth Muscle: Permissive for Maintenance of CRHR2 Expression Xiaoyan Zhu, Shan Wang, Binhai Cong, Xingji You, Xin Ni Second Military Medical University, Shanghai, China Urocortin (Ucn), a member of corticotroping-releasing hormone (CRH) family, has been implicated to be one of the endogenous regulators in cardiovascular system and exerts its effects locally via an autocrine/paracrine fashion. Previous studies have shown the gender difference in CRH-induced vasodilation in human skin, which is related to the concentration of estrogens during menstrual cycle. The aim of this study was to investigate whether estrogens modulate urocortin/corticotroping-releasing hormone receptor type 2 (CRHR2) expressions in vascular smooth muscle, thereby leading to vasodilation. We performed Sham-operation or bilateral ovariectomy (OVX) on female Sprague-Dawley rats. OVX rats were subcutaneously administered with 17b-estradiol (E[sub]2[/sub]) at a dose of 30 [micro]g/kg/day or with placebo for twelve weeks. Primary smooth muscle cells of aorta were used [italic]in vitro[/italic] study. It was found that the urocortin-induced vasodilation and CRHR2 expression were decreased in OVX rats, which were restored by E[sub]2[/sub] replacement treatment for twelve weeks. E[sub]2[/sub] increased the expression of CRHR2 in cultured smooth muscle cells, which were blocked by estrogen receptor b antagonist. Urocortin significantly suppressed the phenylephrine-induced PLCb3 activation, IP3 production and [Ca2+]i elevation. Urocortin stimulated the expression of active GTP-bound Gas protein and cAMP production. The suppressive effects of urocortin on phenylephrin-induced IP3 production and [Ca2+]i elevation were blocked by the inhibitors of adenylate cyclase and PKA. Our results demonstrate that estrogen maintains the expression of CRHR2 in aorta smooth muscle, thereby enhancing vasodilator actions of urocortin. Urocortin exerts its vasorelaxant effects via Gas-cAMP-PKA signaling, leading to down-regulation of PLCb-IP[sub]3[/sub]-Ca[sup]2+[/sup] signaling pathway.[br][br]Sources of Research Support: This work was supported by National Natural Science Foundation of China No. 31000516, No. 30800908 and Science and Technology Commission of Shanghai Municipals (09XD1405600).[br][br]Nothing to Disclose: XZ, SW, BC, XY, XN 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 82 197 1586 SUN-569 PO24-02 Sunday 1383 2012


1380 ENDO12L_SUN-570 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Differential Regulation of Nox1 and Nox4 in Estradiol-17[beta] Mediated Inhibition of Human Vascular Smooth Muscle Cell Proliferation Sigal Shaklai, Meital Grafi-Cohen, Gary Weisinger, Esther Knoll, Ariel Many, Rona Limor, Naftali Stern, Dalia Somjen Sourasky Medical Center, Tel-Aviv, Israel; Tel-Aviv University, Tel-Aviv, Israel [underline]Background[/underline]: The lower incidence of cardiovascular disease (CVD) in premenopausal women than in men of similar age and the menopause-associated increase in CVD has led to the speculation that estradiol-17[beta] (E[sub]2[/sub]) plays a protective role from vascular pathologies. Clinical studies however, have shown equivocal results. The [ldquo]timing hypothesis[rdquo] attempts to settle the discordance, suggesting that E[sub]2[/sub] treatment is protective only in early menopause before accumulation of vascular pathologies. E[sub]2[/sub] is a redox active substance. Reactive oxygen species (ROS) function as second messengers but upon accumulation lead to age related vascular pathologies. Thus, if ROS take part in mediating E[sub]2[/sub] effects on the vasculature, when overplayed or poorly timed, could prove harmful.[br][underline]Aims:[/underline] We hypothesized that E[sub]2[/sub] inhibition of human vascular smooth muscle cell (hVSMC) proliferation, a presumably protective vascular effect, is mediated at least in part by NADPH oxidase (Nox) dependent ROS formation.[br][underline]Methods[/underline]: hVSMCs treated with E[sub]2,[/sub] PPT and DPN (ER[alpha] and ER[beta] agonists respectively) were assessed for ROS using the 2[apos],7[apos]-DCF fluorescent and NBT colorimetric methods, for Nox1 and Nox4 expression by western blot analysis and semi quantative RT-PCR and for proliferation by[sup] 3[/sup][H]-thymidine incorporation.[br][underline]Results:[/underline] ROS production peaked at 1h exposure to E[sub]2, [/sub]PPT and DPN. E[sub]2 [/sub]led to an early transient reduction in Nox1 protein with a concomitant increase in [italic]Nox1[/italic] mRNA. Nox4 protein and mRNA levels were unaffected by treatment. hVSMC proliferation was inhibited after 24h exposure to 30nM E[sub]2, [/sub]DPN and the phytoestrogen Daidzein. Pre-treatment with the general Nox inhibitor DPI and Nox 1/4 specific inhibitor STK, prevented E[sub]2 [/sub]mediated ROS production and inhibition of cell proliferation.[br][underline]Conclusions: [/underline]Our results suggest that Nox derived ROS participate in E[sub]2[/sub] mediated inhibition of hVSMC proliferation. It appears that in hVSMCs[italic] Nox1[/italic] is induced by E[sub]2 [/sub]while[italic] Nox4 [/italic]is unaffected. Our results may further suggest that Nox1 activity is regulated at the protein level through a feedback mechanism in which increased activity leads to early protein degradation with rapid return to baseline via induction of transcription.[br][br]Nothing to Disclose: SS, MG-C, GW, EK, AM, RL, NS, DS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1038 197 1587 SUN-570 PO24-02 Sunday 1384 2012


1381 ENDO12L_SUN-571 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Paxillin Modulates Extranuclear Steroid Signaling and Intranuclear Steroid-Mediated Transcription by Distinct Mechanisms in Prostate Versus Breast Cancer Cells Ismary O De Castro, Aritro Sen, Stephen R Hammes University of Rochester Medical Center, Rochester, NY Extranuclear steroid-mediated kinase signaling modulates cancer cell proliferation, migration and invasion through regulation of gene transcription. However, how this integrative crosstalk between extra- and intra-nuclear steroid signaling is being regulated remains elusive. We have previously shown that the multidomain scaffold protein paxillin serves as a liaison between nuclear and extranuclear signaling in prostate cancer. In fact, paxillin is a critical regulator of androgen and epidermal growth factor (EGF)-induced Erk signaling, and subsequent cell proliferation in prostate cancer cells. In this study, we investigate and compare the regulatory role of paxillin in androgen versus estrogen (E2) actions. In prostate and MCF-7 breast cancer cells, androgens and E2 induce Erk1/2 signaling, respectively and siRNA-mediated knockdown of paxillin abrogates this Erk activation. Furthermore, we demonstrate that paxillin is necessary for androgen receptor (AR) nuclear localization as well as androgen-induced prostate specific antigen (PSA) promoter activity, thereby suggesting that paxillin is regulating PSA gene expression by mediating AR subcellular localization. Interestingly, paxillin also binds to the PSA promoter and likely is acting as a potential co-regulator of AR-mediated PSA transcription. In contrast, paxillin appears to be less crucial for intra-nuclear estrogen receptor (ER[alpha]) actions. For instance, ER[alpha] nuclear localization and E2-triggered ERE luciferase activity in MCF-7 breast cancer cells is independent of paxillin. Similarly, breast cancer cell proliferation is less affected in absence of paxillin. In light of these results, we propose that paxillin is an important mediator of extranuclear steroid signaling but has distinct roles in regulating intranuclear sex steroid signaling. Whether this finding is exclusive for AR and ER[alpha] in other cells is not known. We therefore investigated the impact of paxillin in primary mouse granulosa cells (GC), which are known to express both receptors with well-defined physiology. Intriguingly we find in GCs that paxillin is necessary for AR but not ER[alpha] nuclear localization. In summary, these data suggest that while paxillin is essential for steroid-mediated MAPK signaling, it regulates only AR but not ER-mediated transcription. Importantly, further ongoing studies are aimed at determining other AR and ER-specific genes affected by paxillin in these two important diseases.[br][br]Nothing to Disclose: IODC, AS, SRH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 842 197 1588 SUN-571 PO24-02 Sunday 1385 2012


1382 ENDO12L_SUN-572 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Androgens Decrease Cell Viability in Oxidative Stressed Dopamine Neurons through a Caspase-1 Mediated Mechanism Rosa Lemus, Babak Abbassi, Chang Su, Meharvan Singh, Rebecca L Cunningham University of North Texas Health Science Center, Fort Worth, TX Male gender is one of the clinical risk factors for Parkinson[apos]s disease (PD), a neurodegenerative condition associated with oxidative stress and the loss of dopamine neurons within the nigrostriatal pathway. While the elevated risk in males might point to a potential role of androgens, given the relatively higher levels of androgens in men versus women, the precise role of androgens in PD is unclear. What is known is that androgens can increase the expression of caspase-1, an enzyme whose activity increases with oxidative stress inflammation, and results in apoptosis, mitochondrial collapse, ubiquitination, and a-synuclein-positive Lewy body accumulation. We hypothesize that androgens, through a repression of KLF4 (a negative regulator of caspase-1 expression), results in overexpression of caspase-1, leading in turn to enhanced vulnerability of dopaminergic cells to oxidative damage. To test our hypothesis, we exposed a dopaminergic cell line (N27 cells) to a sublethal concentration of the pro-oxidant, tert-butyl-hydrogen peroxide (H202) for 24 hours followed by exposure to testosterone. Physiologically relevant concentrations of testosterone (0, 1, 10, 100 nM) failed to compromise cell viability in non-oxidatively stressed cells. In contrast, testosterone did promote cell death in the H202 pre-treated cells. In H202-pretreated cells, testosterone increased caspase-1 expression and activation, as assessed by an increase in levels of cleaved caspase-1. Further, testosterone decreased KLF4 expression in H202-treated dopamine cells. The role of KLF4 as a negative regulator of caspase-1 was confirmed in experiments showing that siRNA-mediated knockdown of KLF4 increased caspase-1 levels in H202-treated cells. Notably, testosterone suppressed H202-induced ubiquitination of proteins, supporting a model where androgens, under conditions of oxidative stress, increase the vulnerability of dopamine neurons through a caspase-1 signaling cascade that in turn, induces a dysregulation of the ubiquitin system.[br][br]Sources of Research Support: In part by AHA BGIA4180116 and UNTHSC seed grants to RLC and NIH AG022550 and AG027956 grants to MS.[br][br]Nothing to Disclose: RL, BA, CS, MS, RLC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1988 197 1589 SUN-572 PO24-02 Sunday 1386 2012


1383 ENDO12L_SUN-573 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Androgens Suppress PPAR[gamma] Expression in Human Prostate Cancer Cells Emuejevoke Olokpa, Adrienne Bolden, LaMonica Vanette Stewart Meharry Medical College, Nashville, TN The peroxisome proliferator activated receptor gamma (PPAR[gamma]) is a ligand-activated transcription factor that plays a key role in adipocyte differentiation, insulin sensitivity, and lipid metabolism. PPAR[gamma] also appears to function as a tumor suppressor. Conditional knockout of PPAR[gamma] within the mouse prostate results in development of prostate intraepithelial neoplasia (PIN), a precursor to prostate cancer. In addition, ligands that activate this receptor have been shown to suppress growth of cultured human prostate cancer cells and prostate cancer xenografts [italic]in vivo[/italic]. The factors that control PPAR[gamma] expression within prostate cancer cells have not been characterized. The androgens testosterone and dihydrotestosterone (DHT) reduce expression of PPAR[gamma] protein within adipose tissue. PPAR[gamma] protein levels are higher in prostate cancer cells that lack the androgen receptor (AR). However, it was not known if androgen-mediated activation of AR also controls PPAR[gamma] levels or PPAR[gamma] function in early or late stage prostate cancers. The goal of this study was to examine whether androgens regulate expression of PPAR[gamma] within human prostate cancer cells. To accomplish this goal, we first explored the effect of androgens on PPAR[gamma] protein levels within the AR positive, androgen-independent C4-2 prostate cancer cell line. Western blot analyses revealed that DHT reduced nuclear levels of PPAR[gamma] protein in C4-2 cells in a dose-dependent manner. The greatest decrease in PPAR[gamma] occurred when C4-2 cells were exposed to DHT concentrations [ge] 1 nM. Decreases in PPAR[gamma] protein could be detected after six hours of DHT exposure, and the levels remained low after treatment for twenty-four hours. This response was not unique to C4-2 cells, for we also saw a decrease in PPAR[gamma] protein levels in LNCaP cells following DHT treatment. Concentrations of DHT that reduced PPAR[gamma] protein levels were effective at lowering PPAR[gamma] mRNA levels within C4-2 cells. Therefore, the androgen-induced reduction in PPAR[gamma] levels appears to be due in part to a reduction in PPAR[gamma] mRNA levels. While androgens suppressed PPAR[gamma] within LNCaP and C4-2 cells, they did not alter PPAR[gamma] levels in the AR-negative PC-3 prostate cancer cell line. In addition, in androgen-replete media siRNA-mediated knockdown of AR reduced PPAR[gamma] protein levels within C4-2 cells. In conclusion, our data suggest androgens do regulate expression of PPAR[gamma] in AR-positive prostate cancer cells and that this response requires the presence of AR.[br][br]Sources of Research Support: NIH Grant CA114253 awarded to LVS.[br][br]Nothing to Disclose: EO, AB, LVS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 363 197 1590 SUN-573 PO24-02 Sunday 1387 2012


1384 ENDO12L_SUN-574 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) The Effects of Sex Steroids on Cardiac Iron Loading in a Mouse Model of Iron Overload Casey J Brewer, Maya Otto-Duessel, Ruth I Wood, John C Wood University of Southern California, Los Angeles, CA; City of Hope, Duarte, CA; Children[apos]s Hospital Los Angeles, Los Angeles, CA Background: Thalassemia is the most common genetic disease worldwide with a very high prevalence in Asia (1). Patients with thalassemia major require chronic blood transfusions to treat their severe anemia. While this treatment corrects the patients[apos] anemia, it leads to a severe iron overload that can become lethal in their second decade of life (2). The leading cause of death in these patients is cardiomyopathy due to iron overload of the heart (3). Women with thalassemia have a 2:1 survival advantage compared to men with the disease (4). The question is raised whether sex steroids influence cardiac iron loading. This idea is supported by data from our lab showing that male mice load more iron into their hearts than female mice. Methods: At four weeks of age, female mice were ovariectomized (OVX) and male mice were castrated (ORCHX). Female mice received either an estrogen implant or a cholesterol control; male mice received an implant containing testosterone, dihydrotestosterone (DHT), estrogen, or cholesterol. The mice were placed on a high iron diet for the next eight weeks. At twelve weeks of age the mice were sacrificed and the heart and liver tissue were harvested. Tissue iron concentrations were determined by atomic absorption. Results: When a pool of all male mice (ORCHX, ORCHX+T, ORCHX+E, ORCHX+C, and ORCHX+DHT) were compared to a pool of all female mice (OVX and OVX+E), the pooled males had significantly more cardiac iron than pooled females (p[lt]0.0001). This suggests an organizational effect of steroids on cardiac iron loading. In females, OVX+E mice did not have a significant difference in cardiac iron than OVX. In males, ORCHX+E mice did show a statistically significant increase in cardiac iron vs. ORCHX (p[lt]0.02). ORCHX+T displayed an upward trend in cardiac iron compared to ORCHX (p = 0.11), but this was not statistically significant. Conclusions: There is an organizational effect of steroids on cardiac iron loading, predisposing male mice to higher levels of cardiac iron than females. Estrogen also has an activational effect of increasing cardiac iron loading, but only in males.[br][br]1) Weatherall, D. Am J Hum Genet 2004 March; 74(3):385[ndash]392. 2) Wood JC et al., Haematologica 2008; 93(6):917[ndash]920. 3) Wood JC et al., Blood 2010; 116:537-543. 4) Borgna-Pignatti C et al., Haematologica 2004; 89(10):1187-1193.[br][br]Nothing to Disclose: CJB, MO-D, RIW, JCW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 686 197 1591 SUN-574 PO24-02 Sunday 1388 2012


1385 ENDO12L_SUN-575 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) The Role of Testosterone in the Control of Muscle Protein Balance Lucian Nicolae Sandor, Daniel Lee, Gianluca Toraldo, Anqi Zhang, Ravi Jasuja, Shalender Bhasin, Carlo Serra Boston Medical Center, Boston, MA Background: Skeletal muscle is the main protein and energy storage of the body. Aging and pathological conditions perturb protein turnover and the muscle response to anabolic stimuli. Gains in muscle mass occur through either increases in muscle protein synthesis or declines in muscle protein breakdown. Several lines of evidence indicate that testosterone increases muscle mass by stimulating either protein synthesis and reducing protein degradation. In addition, androgens prevent muscle atrophy and weakness caused by glucocorticoids in humans and rodents. These data suggest that androgens inhibit proteolysis, and enhance specific steps of protein synthesis and/or protect from degradation key factors of the translational machinery.[br]Methods: To assess the effect of testosterone on muscle protein synthesis and degradation we used the Hershberger assay in rodents. 2-month old male mice were castrated and treated with vehicle or testosterone propionate. Mice were killed after 4 and 7 days of testosterone treatment. Sham-operated, untreated mice and 7-day castrated mice served as control.[br]Results: The androgen-dependent [italic]levator ani[/italic] muscle underwent rapid atrophy after castration, showed reduced level of androgen receptor and FoxOs phosphorylation, as well as increased MAFbx and MuRF1 gene expression. Testosterone administration rescued these changes. Castration induced the expression of key autophagy genes, such as Bnip3, LC3B, p62 and Cathepsin-L, indicating an active recruitment of autophagy components during [italic]levator ani[/italic] muscle atrophy. Castrated mice showed also the increased gene expression of Fis1, a component of the mitochondrial fission machinery activated during autophagy. Castration was associated to reduced activation and activity of mTOR, as indicated by the level of its phosphorylation and by that of 4E-BP1. Testosterone rescued these effects and enhanced the phosphorylation of eIF4G and of the ribosomal protein S6rp, a target of activated p70[sup]S6k1[/sup]. Castrated mice showed reduced protein level of eIF3f, of eIF2B[epsilon], the enzymatic subunit of the eIF2B complex, and of the elongation factor eEF2. Testosterone rescued all these changes.[br]Conclusion: Testosterone supplementation re-establishes an optimal protein balance in atrophic muscle, and regulates the protein degradation pathways as well as the early steps of mRNA translation. These data indicate potential new therapeutic targets in androgen-based therapies for muscle wasting.[br][br]Nothing to Disclose: LNS, DL, GT, AZ, RJ, SB, CS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 272 197 1592 SUN-575 PO24-02 Sunday 1389 2012


1386 ENDO12L_SUN-576 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Enhanced Evaluation of Selective Androgen Receptor Modulators [italic]In Vivo[/italic] Maya Otto-Duessel, Miaoling He, Jeremy O Jones City of Hope Beckman Research Institute, Duarte, CA [bold][underline]Introduction:[/underline][/bold] The androgen receptor (AR) mediates the response to testosterone and dihydrotestosterone (DHT). AR exhibits different activity in different tissues. Changes in AR activity are associated with many diseases, including prostate cancer. Treating these diseases by either systemically blocking or increasing AR activity causes severe side-effects due to changes of AR activity in non-diseased tissues. Patients in need of treatment for AR-dependent diseases would benefit from therapies that target AR activity in specific tissues. A new class of drugs called selective AR modulators (SARMs) may provide the desired selective effect. However, we lack adequate models for evaluating potential SARMs [italic]in vivo[/italic]. Therefore, we created a profile of androgen responsive genes in relevant AR expressing tissues. We used this panel of genes to compare transcriptional changes in response to DHT and the SARM bolandiol at 16h and 6wks. We also compared these transcriptional changes to the physiological changes traditionally used to assess selectivity in the Hershberger assay.[br][bold][underline]Methods:[/underline][/bold] Forty eight male rats were divided into 4 groups; intact, castrate, castrate + DHT, and castrate + bolandiol, and treated for either 16h or 6wks. After treatment, animals were euthanized and organs harvested, weighed and processed for QRT-PCR of androgen-responsive genes. For the 6wk experiment, tibia and femur were processed for microCT scans to assess bone density and serum lipids as well as testosterone levels were determined by biochemical assays.[br][bold][underline]Results: [/underline][/bold]We identified known and novel androgen-regulated genes in prostate, muscle, bone, brain, skin, fat, and kidney. We were able to identify genes whose expression after 6wks of treatment correlated with the corresponding physiological endpoints. In the muscle for instance, treatment with bolondial for 6wks was 74% and 79% as effective as DHT treatment in stimulating the transcription of [italic]amd1[/italic] and [italic]acta1[/italic]. Also, bolandiol treated muscles weighed 79% of DHT treated ones. Furthermore, the expression of many of these androgen-regulated genes was similar whether measured after 16hrs or 6wks of treatment, suggesting that early transcriptional changes could be used as a surrogate for later physiological changes.[br][bold][underline]Conclusion[/underline][/bold][underline]:[/underline] Our findings suggest that measuring the transcription of androgen-regulated genes offers a rapid and quantifiable addition to the conventional Hershberger assay. However, larger studies are needed to confirm these findings.[br][br]Nothing to Disclose: MO-D, MH, JOJ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 361 197 1593 SUN-576 PO24-02 Sunday 1390 2012


1387 ENDO12L_SUN-577 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Identification of Four New Sex Hormone-Binding Globulin Variants with Abnormal Production or Function Tsung-Sheng Wu, Caroline Underhill, Geoffrey Lewis Hammond Child [amp] Family Research Institute, Vancouver, Canada [bold]Background: [/bold]Sex hormone-binding globulin (SHBG) binds sex steroids with high affinity and specificity. The levels of SHBG in blood determine the non-protein bound [ldquo]free[rdquo] fractions of sex steroids that are able to access target tissues and cells. Genetic polymorphisms of [italic]SHBG[/italic] have been associated with altered SHBG levels or functions and linked to the risk of several diseases, including reproductive tissue cancers and type 2 diabetes. Recently, large scale genome and exome sequencing projects have revealed numerous single nucleotide polymorphisms (SNP) within the [italic]SHBG[/italic] gene. Based on our knowledge of SHBG tertiary and quaternary structure, we selected nine new SNPs, which result in amino acid substitutions in the SHBG amino-terminal LG domain that mediates steroid-binding and dimerization, and examined their impact on SHBG production and/or function.[br][bold]Methods:[/bold] SHBG variants were expressed in Chinese hamster ovary cells for characterization by steroid-binding capacity and time-resolved immunofluorometric assays, which allow the steroid binding and dimerization properties of SHBG to be assessed.[br][bold]Results:[/bold] Four out of the nine naturally occurring SHBG variants studied have specific abnormalities. SHBG T48I and SHBG R123H have reduced steroid-binding affinity; SHBG R154W is dimerization deficient and has a lower binding affinity for estrogens but a normal affinity for androgens; SHBG G195E is not produced or secreted efficiently, probably due to misfolding during synthesis.[br][bold]Conclusions: [/bold]A method for the screening and functional analysis of SHBG genetic variants has been implemented to assess the impact of newly discovered genetic variants of SHBG on its production and/or function. This information will need to be assessed in relation to a wide range of diseases that have already been associated with [italic]SHBG[/italic] polymorphisms. In addition, algorithms designed to calculate free testosterone or estradiol levels will need to take into consideration the effects of these new SHBG variants.[br][br]Sources of Research Support: Canadian Institutes of Health Research grant MOP-15261 awarded to GLH.[br][br]Nothing to Disclose: T-SW, CU, GLH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2218 197 1594 SUN-577 PO24-02 Sunday 1391 2012


1388 ENDO12L_SUN-578 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Oxidative Stress Negatively Influences the Effects of Androgens on Dopamine Neuronal Viability Babak Abbassi, Meharvan Singh, Rebecca L Cunningham University of North Texas Health Science Center, Fort Worth, TX Parkinson[apos]s disease (PD) is a progressive neurodegenerative disorder, in which aged men have a greater incidence than women. PD is characterized by oxidative stress and dopamine neuronal loss in the nigrostriatal pathway. The mechanisms underlying this gender bias remain elusive, although one possibility may be that oxidative stress converts the neuronal response to androgens to that which is toxic. Specifically, we hypothesize androgens, such as testosterone, compromise dopamine neuron viability only in an oxidative stress condition. To test our hypothesis, we exposed a dopaminergic cell line (N27 cells) to a sublethal concentration of the pro-oxidant, tert-butyl-hydrogen peroxide (H202) for 24 hours and assessed cell viability in the presence or absence of testosterone. Physiologically relevant concentrations of the androgen, testosterone (0, 1, 10, 100 nM) failed to compromise cell viability in non-oxidatively stressed cells. In contrast, testosterone and the membrane impermeable androgen, testosterone conjugated to BSA (T-BSA), did promote cell death in the H202 pre-treated cells. Interestingly, androgen pre-treatment protected dopamine cells from H202-induced cell death. Supporting the role of oxidative stress as a switch in this effect, the antioxidant, N-acetyl cysteine, prevented the damage promoting effects of testosterone in H202 pretreated cells. Neither the androgen receptor nor the estrogen receptor antagonists, flutamide (10 uM) and ICI 182, 780(1 uM), respectively, altered the death promoting effect of testosterone. Coupled with the observation that T-BSA mimicked the effects of testosterone, we suggest that the cell death promoting effects may be mediated by a putative membrane-associated androgen receptor. Together, these results indicate that oxidative stress acts as a molecular switch in dopamine neurons that can reverse the neuroprotective effects of androgens to that which is neurotoxic. Thus, the interplay between oxidative stress and androgens on dopamine neuronal viability may underlie the male gender bias found in PD.[br][br]Sources of Research Support: In part by NIH NS061417 and AHA BGIA4180116 grants to RLC and NIH AG022550 and AG027956 grants to MS.[br][br]Nothing to Disclose: BA, MS, RLC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2167 197 1595 SUN-578 PO24-02 Sunday 1392 2012


1389 ENDO12L_SUN-579 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Testosterone Protects Cardiomyocytes from Superoxide Injury Via NF-kB Signaling Pathways Paul A Komesaroff, Fuying Xiao, Lina Nheu, Shanhong Ling Monash University, Melbourne, Australia Deficiency of testosterone (T) in both ageing men and in women after oophorectomy is associated with increased risk of cardiovascular disease, including myocardial infarction, and in these cases there is evidence that T therapy may provide cardiac protection. To date, however, the cellular and molecular mechanisms underlying the actions of T on the heart remain unclear. Signalling pathways of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-[kappa]B), and particularly the canonical (RelA/p50 dimer) NF-[kappa]B pathway, have been recognised to play key roles in the regulation of apoptosis and necrosis, hypertrophy, myocardial recovery and pathological remodelling of the heart. We therefore examined the impact of exogenous T on the viability of cardiac myocytes through the NF-[kappa]B signalling pathway. Rat (H9c2) cardiomyocytes were cultured in 10% FBS-DMEM and incubated with hydrogen peroxide (H2O2, 100[mu]M, 6 hrs) in the presence or absence of T (50-100nM, 24 hrs). NF-[kappa]B expression was knocked down by a RelA (p65)-specific siRNA and by blockage of the classical androgen receptor (AR) by flutamide (100nM). The study found that: (1) T administration was associated with a reduction in superoxide-induced apoptotic/necrotic death in normal myocytes that was partly blocked by flutamide but which was completely absent in the NF-[kappa]B-knockdown myocytes; (2) T stimulated NF-[kappa]B (RelA) expression in normal but not in knockdown myocytes, and this effect was blocked by flutamide; (3) T activated ERK1/2 and PKC activities, with no change with flutamide or knockdown of NF-[kappa]B; (4) expression of caspase-3 increased in the NF-[kappa]B knockdown myocytes, and this appeared to be reduced by T via an AR-mediated pathway; and (5) Akt was significantly activated by oxide stress in normal but not NF-[kappa]B-knockdown myocytes, an effect which was enhanced by T administration. In conclusion, exogenous T supplementation protects cardiomyocytes from oxide stress injury via the canonical NF-[kappa]B signalling pathway, possibly by activating cytosol NF-[kappa]B dimer release via non-AR-mediated ERK1/2 and PKC activation or by stimulating nuclear NF-[kappa]B expression via AR mediation, resulting in a reduction of caspase-3 expression and an enhancement of Akt activation.[br][br]Nothing to Disclose: PAK, FX, LN, SL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1067 197 1596 SUN-579 PO24-02 Sunday 1393 2012


1390 ENDO12L_SUN-580 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Virus Infection Potentiated Glucocorticoid-Induced Interleukin-10 Production in Dendritic Cells by Phosphorylating the Glucocorticoid Receptor through Activation of the Extracellular Signal-Regulated Kinase 1/2: Potential Implication to Stress-Mediated Modification of Susceptibility to Viral Infection Sinnie Sin Man Ng, Andrew Li, Ilia J Elenkov, George N Pavlakis, Ozato Keiko, Tomoshige Kino National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD; The Chinese University of Hong Kong, Shatin, China; Italian National Research Council, Roma, Italy; National Cancer Institute at Frederick, NIH, Frederick, MD; National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD Glucocorticoids (GCs) are end-products of the hypothalamic-pituitary-adrenal axis, which plays a central role in the adaptive response to stress including infection with pathogens, while the glucocorticoid receptor (GR), an intracellular receptor of the nuclear receptor superfamily, mediates diverse cellular actions of these hormones. It is known that physical and/or mental stress alters the susceptibility to viral infection, but the cellular targets and mediators underlying this phenomenon are poorly understood. We previously reported in this meeting that infection of the Newcastle disease virus (NDV) synergistically increased GC-stimulated secretion of the anti-inflammatory cytokine interleukin (IL)-10 by 7 (mRNA) and 3 (protein)-fold in murine dendritic cells (DCs), the versatile and central components of host immune system by sensing antigens and by stimulating downstream immune components. We further examined the molecular mechanisms underlining this synergism: We found that the synergism was caused by activation of the signal(s) downstream of the Toll-like receptor (TLR) 4, because lipopolysaccharide (LPS), a specific TLR4 activator, also enhanced GC-induced IL-10 mRNA expression similar to NDV infection. With the screening using several chemical inhibitors that block various intracellular signaling pathways downstream of TLR4 for IL-10 expression, the compound U126, an inhibitor of the extracellular signal-regulated kinase (ERK) 1/2, attenuated NDV-induced enhancement of dexamethasone (DEX)-stimulated IL-10 mRNA/protein expression. Overexpression of ERK1/2 or the transdominant mutant ERK2 D319N enhanced DEX-induced transcriptional activity of the IL-10 promoter in the presence of the wild type human GR in HCT116 colon carcinoma cells, while ERK2 failed to enhance the DEX effect on this promoter in the presence of the mutant human GR defective in serine at amino acid position 203, but not the GR defective in serine 211 or 226. Taken together, these results indicate that viral infection enhances GC-induced production of IL-10 in DCs by activating ERK1/2 through TLR4, and further, by phosphorylating GR at serine 203. Our results suggest that virus increases its infectivity and invasion to host tissues through cooperated expression of IL-10 with GCs in these key immune cells. Further, they may partly explain why stress and GCs increase viral infection-caused exacerbation of allergic disorders, such as asthma.[br][br]Sources of Research Support: This study was funded partly by the [italic]Eunice Kennedy Shriver[/italic] National Institute of Child Health and Human Development and the National Cancer Institute at Frederick, National Institutes of Health.[br][br]Nothing to Disclose: SSMN, AL, IJE, GNP, OK, TK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 177 197 1597 SUN-580 PO24-02 Sunday 1394 2012


1391 ENDO12L_SUN-581 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Nandrolone, an Anabolic Steroid, Induces the Expression of Numb, a Notch Inhibitor, Via Activation of Wnt Signaling: A Novel Mechanism of Crosstalk between Wnt and Notch Signaling in C2C12 Myoblasts Xin-Hua Liu, Jiangping Pan, Yong Wu, Weiping Qin, Lauren Collier, William A Bauman, Christopher Cardozo James J Peter Veteran Affairs Medical Center, Bronx, NY; Mount Sinai School of Medicine, New York, NY; Mount Sinai School of Medicine, New York, NY Wnt signaling is essential for myogenic differentiation. Activation of Wnt signaling results in the transition of progenitors from the proliferation phase to the differentiation phase during myogenesis. In this process, Wnt signaling directs a critical transition from high to low Notch activity. We have reported that in denervated rat muscle nandrolone, an anabolic steroid, prevented upregulation of Notch signaling and increased the expression of Numb, a Notch inhibitor and a critical regulator of myogenic differentiation. We also demonstrated in C2C12 cells that nandrolone increased Numb protein levels. With these considerations in mind, we investigated the effects of nandrolone on Wnt activity and determined the interactive role of Wnt signaling in nandrolone-induced Numb expression in C2C12 cells. When the cells were cultured in a differentiation-favorable medium (MDM+2% horse serum), nandrolone promoted nuclear accumulation of [beta]-catenin, with a peak effect noted at 48h after the treatment. Nandrolone also increased the activity of a Tcf-luciferase reporter indicating enhanced activity of Tcf transcription factors. In addition, nandrolone increased GSK3[beta] phosphorylation at Ser9, which inhibits GSK3[beta] activity and promotes nuclear entry of [beta]-catenin. To determine whether nandrolone-induced Wnt signaling activation is necessary for its effect on Numb upregulation, the effects of naturally occurring inhibitors of Wnt signaling were tested on the expression of Numb. Pre-treatment of cells with DKK1 or sFRP1 significantly inhibited basal and nandrolone-induced expression of Numb mRNA and protein. Conversely, incubation of cells with Wnt3a resulted in an upregulation of Numb mRNA/protein and diminished nandrolone-induced increases in Numb levels. To extend these findings, [beta]-catenin siRNA was employed. Cells that were transfected with [beta]-catenin-siRNA had decreased Numb mRNA and protein levels compared to those transfected with scrambled siRNA (negative control). Furthermore, SB216763, a GSK3[beta] inhibitor, upregulated [beta]-catenin levels, and increased Numb mRNA and protein expression in the same cells. Finally, CHIP assay demonstrated that nandrolone promoted the recruitment of [beta]-catenin to the promoter region of the Numb gene. These results indicate a role for Wnt signaling in nandrolone-induced Numb expression and reveal a novel mechanism by which nandrolone induces a critical interaction between Wnt and Notch signaling pathways during myogenesis.[br][br]Sources of Research Support: This work was supported by the Veterans Health Administration, Rehabilitation Research and Development Service (Grants B4162C, F7756R and F6997R).[br][br]Nothing to Disclose: X-HL, JP, YW, WQ, LC, WAB, CC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 364 197 1598 SUN-581 PO24-02 Sunday 1395 2012


1392 ENDO12L_SUN-582 POSTER SESSION: Steroid Hormone Action: Genomic [amp] Non-Genomic Signaling (1:30 PM-3:30 PM) Effect of Progesterone on Melanoma Cell Growth Pandurangan Ramaraj, James L Cox KCOM, AT Still University, Kirksville, MO Sex steroids such as Androgens, Estrogen, Progesterone have roles in maintaining a healthy skin. Imbalances in these hormones result in skin diseases. One of the deadliest form of skin diseases is Melanoma. Only scanty information is available on the effect of sex steroids on melanoma. So we decided to check the effect of Cholesterol (Chol), dehydroepiandrosterone (DHEA), Androstenedione (AD), Testosterone (T) and Progesterone (P) on melanoma cell growth.[br]B16 mouse melanoma cells were grown in RPMI medium. Test hormones were added to the medium at 100, 150 and 200 [micro]M concentrations. After 48 hours viable cells were assessed by MTT assay. Of the five steroids tested, progesterone showed a significant inhibition in cell growth (87% at 200[sub] [micro][/sub]M concentration). Further assay with progesterone showed a dose-dependent decrease in B16 cell growth. Similar dose-curve pattern was observed with human melanoma (BLM) cells. Time-course study with BLM cells revealed that 48 hrs and 72 hrs of incubation yielded similar pattern. Progesterone decreased cell growth by inducing cell death. So we investigated the mechanism of cell death. Cell death by necrosis was ruled out by trypan blue dye exclusion test. Apoptosis was also ruled out by DAPI nuclear staining and lack of DNA ladder by agarose gel analysis of DNA from progesterone treated cells. Finally, autophagy as the cause of cell death was determined by incubating the cells with 3-methyl-Adenine (3-MA). Addition of 3-MA (2 mM) along with progesterone rescued the cells partially from cell death. Hence, decided to block the action of progesterone (10 [micro]M) by co-incubating cells with RU-486 (10, 50 100 [micro]M concentrations), an antagonist to progesterone receptor. But co-incubation experiment showed an additive effect at P-10[sub] [micro][/sub]M and RU-50[sub] [micro][/sub]M concentration itself, producing a maximum inhibition (71%) of cell growth. Pre-incubation of cells with progesterone (1-10 [micro]M), before adding 10, 50 and 100 [micro]M concentrations of RU-486, did not show any significant change from the usual RU-486 treatment pattern. Similarly, pre-incubation of cells with 1[micro]M of RU-486 before adding progesterone, did not show any significant change from the usual progesterone treatment pattern, suggesting progesterone actions might not be mediated through its receptor.[br]In conclusion, progesterone decreased melanoma cell growth by inducing autophagy and this action was not mediated through progesterone receptor.[br][br]Nothing to Disclose: PR, JLC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 93 197 1599 SUN-582 PO24-02 Sunday 1396 2012


1393 ENDO12L_SUN-588 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Adult Final Height in GH-Treated Children with GH Deficiency Corina Galesanu, Andra Loghin, Petronela Aancute, Luminita Apostu, Mihail Romeo University of Medicine and Pharmacy [quot]Gr T Popa[quot], Iasi, Romania; Centre of Imaging and Radiological Diagnosis, Iasi, Romania Introduction: Both GH and IGF-1 are essential for normal growth in children. Bone age (BA) in children with short stature and GH-deficiency (GHD), is known to be delayed. Various factors have been identified that influence the growth response for GH-treatment in children with GHD. For the patients are very important to obtain a normal or near-normal final adult height.[br]Aims: To investigate the effect of rhGH - treatment in children with GHD to reanch the adult hight.[br]Patients and Methods: We studied 37 children with idiopathic isolated GHD (22 males, 15 females). At start of rhGH-therapy, the mean chronological age (CA) was 9.67[plusmn]3.3 years for the boys and 7.4[plusmn]2.1 years for the girls. All children were prepubertal. The GH dose was 0.3 mg/kg/wk. The mean duration of GH-therapy was 4.2[plusmn]3.3 years. Effectiveness of GH-treatment was documented by height velocity from 3.81 cm/yr to 6.53 cm/yr for the boys, and from 2.94 cm/yr to 5.99 cm/yr for the girls, and also serum IGF-1 levels from 187.85[plusmn]28 to 444.6[plusmn]28 ng/mL. The BA was delayed with 2.03[plusmn]0.21 years at start and the delay decreased at 1.47[plusmn]0.21 year under the GH-treatment. Three of our children - boys started the puberty. Bone age (Greulich and Pyle) advanced and predicted final adult height.[br]Results: The average period of treatment for these three boys was 8.21[plusmn]1.89 yr. In the beginning the mean difference between BA and CA was more then three years and under the treatment the bone age advanced, height velocity was 6.98 cm/yr. In the puberty period with the some dose of rhGH, the height velocity increased to 9.32 cm/yr. Final mean adult height was 170.6[plusmn]4.0 cm, overlapping with normal mean height for our male population 173.3[plusmn]6.2 cm.[br]Conclusion: In our series of children with GHD, treated with rhGH, they reached their normal adult height. The other ones are under the treatment. After the end of linear growth, the adult with GHD must to keep on the treatment in adult life to improuve the quality of life, bone and muscle mass, diminution cardiovascular risk.[br][br]Nothing to Disclose: CG, AL, PA, LA, MR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 195 198 1600 SUN-588 PO29-02 Sunday 1397 2012


1394 ENDO12L_SUN-589 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Single Nucleotide Polymorphisms (SNPs) Associated with Growth Response after 3 Years on Growth Hormone (GH) Therapy in Children with GH Deficiency (GHD) Pierre Chatelain, Eirik Vangsoy-Hansen, Jovanna Dahlgren, Armand Valsesia, Benoit Destenaves, Peter Clayton Universit[eacute] Claude Bernard, Bron, France; Haukeland University Hospital, Bergen, Norway; G[ouml]teborg University, G[ouml]teborg, Sweden; Merck Serono SA, Geneva, Switzerland; Merck Serono SA, Geneva, Switzerland; University of Manchester, Manchester, UK Background:[br]The PREDICT follow-up study investigates the effect of single nucleotide polymorphisms (SNPs) in growth- and metabolism-associated genes on long-term growth in children with GHD on GH therapy.[br]Objective:[br]To evaluate the association of these SNPs with growth response (cm/yr) in Years (Y) 1 to 3 of GH therapy in children with GHD.[br]Methods and subjects:[br]Treatment-na[iuml]ve children with classic idiopathic GHD (Y1, n=110; Y2, n=93; Y3, n=73) were prepubertal at start of GH therapy and Tanner stage 1 and 2 at Y3. Median GH doses were 35, 33 and 32 [sup][mu][/sup]g/kg/day at Y1, Y2 and Y3, respectively. A total of 1451 SNPs from 98 candidate genes were genotyped. Associations of genotypes with growth (cm) over Y3 and changes from baseline to Y3 were assessed in continuous analyses by Kruskal[ndash]Wallis nonparametric tests. SNPs with an adjusted p-value less than 0.05 were considered significant.[br]Results:[br]SNPs in five genes ([italic]AKT2[/italic], [italic]GAB1[/italic], [italic]PROP1[/italic], [italic]SLC2A1[/italic] and -[italic]A4[/italic]) were associated with growth over Y3, while SNPs in seven genes ([italic]FOS[/italic], [italic]GRB2[/italic], [italic]JAK2[/italic], [italic]PI3KR1[/italic] and -[italic]R2[/italic], [italic]SHC1[/italic], [italic]SLC2A4[/italic]) were associated with growth over the first 3 years. SNPs in [italic]GRB2[/italic] and [italic]PIK3R2[/italic], and in [italic]FOS[/italic] and [italic]GRB2[/italic], were associated with growth over Y2 and over the first 2 years, respectively. None of these genes had previously been identified as associated with first-year growth response, although the latter did include [italic]GRB10[/italic].[br]Conclusion:Our results suggest that in GHD the genes associated with the maintenance phase of growth on GH therapy are generally different from those involved in catch-up growth over the first year of treatment. Growth factor receptor-bound proteins are, however, involved in both phases.[br][br]Sources of Research Support: Merck Serono S.A. - Geneva, Switzerland, a branch of Merck Serono S.A., Coinsins, Switzerland, an affiliate of Merck KGaA, Darmstadt, Germany.[br][br]Disclosures: PC: Consultant, Merck Serono S.A.; Investigator, Merck Serono S.A. JD: Speaker, Merck Serono S.A.; Consultant, Merck Serono S.A. AV: Employee, Merck Serono S.A. BD: Employee, Merck Serono S.A. PC: Investigator, Merck Serono S.A.; Speaker, Merck Serono S.A. Nothing to Disclose: EV-H 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 603 198 1601 SUN-589 PO29-02 Sunday 1398 2012


1395 ENDO12L_SUN-590 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) A Network Analysis of Gene Expression through Childhood Highlights Changes Related to Both Age and Growth Adam Stevens, Andrew Whatmore, Benoit Destenaves, Pierre Chatelain, Peter Clayton University of Manchester, Manchester, UK; Merck Serono SA, Geneva, Switzerland; H[ocirc]pital M[egrave]re-Enfant [ndash] Universit[eacute] Claude Bernard, Lyon, France [bold]Objective:[/bold] to assess age- and growth-dependent gene expression in children and correlate this with biological pathways.[br][bold]Methods:[/bold] We conducted a gene expression meta-analysis on datasets from normal children curated from the NCBI Gene Expression Omnibus (GEO). Four datasets were combined to form a group of 87 individuals ranging from 0.2 to 29.3 years of age (average 7.7+/-6.9yr). Analysis of gene expression data was performed using hierarchical clustering and transcription factors involved in the regulation of differentially expressed genes were predicted using Molecular Signatures Database (MSigDB), the DIRE algorithm [Distant Regulatory Elements of co-regulated genes (1)] and Ingenuity Pathways Analysis (IPA) software. Network analysis was used to identify key biological pathways and control points within them.[br][bold]Results:[/bold] In these normal children, 927 gene expression probes were significantly associated with age (adjusted p-value, q[lt]0.1). These probes formed three clusters correlating with the different stages of human growth: 1) [le]6yrs [infancy, early childhood] (408 probes); 2) [gt]6 to [le]17yrs [late childhood, puberty] (252 probes) and 3) [gt]17yrs [adulthood, final height] (267 probes)[br]Two probes represent genes previously identified as functional candidates in a meta-analysis of genetic data associated with human height (2); [italic]LTBP1[/italic] (q[lt]0.05) and [italic]IGF2BP3[/italic] (q[lt]1x10[sup]-17[/sup]).[br]Network analysis identified pathway [ldquo]bottle-necks[rdquo] as indicators of regulatory function (3) and [ldquo]sub-clustering[rdquo] of networks demonstrated enrichment for growth and development gene ontology. The most significant age related changes occurred in NOTCH, VEGF, TGFB and WNT pathways. Analysis of canonical pathways and transcription factors associated with inferred protein interaction networks demonstrated involvement of the glucocorticoid receptor (GR) pathway. Both the infancy and puberty groups showed a change in gene expression in a higher proportion of the GR pathway (+1.8 fold) than the adult group.[br][bold]Conclusion:[/bold] These data identify age-dependent genes that cluster into key growth and development pathways. Changes in these genes may represent potential confounders when examining gene expression profiles associated with both normal growth pattern during childhood and diseases (including treatment) that present either at specific ages such as retinoblastoma and neuroblastoma or at any time through childhood such as diabetes mellitus, asthma or rheumatoid arthritis.[br][br](1). Gotea, V. and I. Ovcharenko. DiRE: identifying distant regulatory elements of co-expressed genes. Nucleic Acids Research 2008; W133-9. (2). Lango AH, Estrada K, Lettre G et al. Hundreds of variants clustered in genomic loci and biological pathways affect human height. Nature 2010; 467(7317):832-838. (3). Yu H, Kim PM, Sprecher E, Trifonov V, Gerstein M. The importance of bottlenecks in protein networks: correlation with gene essentiality and expression dynamics. PLoS Comput Biol 2007; 3(4):e59.[br][br]Disclosures: PC: Consultant, Merck Serono S.A.; Investigator, Merck Serono S.A. Nothing to Disclose: AS, AW, BD, PC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 825 198 1602 SUN-590 PO29-02 Sunday 1399 2012


1396 ENDO12L_SUN-591 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Growth Characteristics and GH-IGF Axis Status in Idiopathic Short Stature (ISS) Patients in the EPIGROW Study Carrying Missense Single Nucleotide Polymorphisms (SNPs) in Growth-Related Genes Claire Bonshek, Mireille Bonnemaire, Pascale Dutailly, Laurent Naudin, Emmanuel Pham, Peter Clayton University of Manchester, Manchester, UK; Ipsen Pharma SAS, Boulogne-Billancourt, France [bold]Background:[/bold] EPIGROW, a prospective epidemiological study conducted in 9 European countries, was established to identify the prevalence of IGF-I deficiency in ISS and to identify genetic SNPs that may be potentially damaging and contribute to the phenotype. 263 prepubertal ISS children (Height SDS [lt]-2, peak GH [ge]7[micro]g/L, no defined aetiology for their SS) were analysed. 53% were found to be IGF-I deficient (SDS [lt]-2). Focussing on known growth disorder genes (GH-IGF axis: [italic]GH-1, GHR, STAT5b, IGF-1R, IGFALS[/italic], and primordial growth (3-M): [italic]CUL7, OBSL1[/italic]) and using next generation deep resequencing, 63 novel and 2 known missense SNPs in 66 children, both as homozygotes and heterozygotes, were identified. These SNPs were present in the cases and not in 263 matched controls.[br][bold]Aim:[/bold] To determine whether there is a difference in growth phenotype and GH-IGF biochemical status for each missense SNP, for homozygotes versus heterozygotes and between those with GH-IGF axis gene SNPs versus those with primordial growth gene SNPs.[br][bold]Methods:[/bold] Growth parameters evaluated included height, weight, sitting height, head circumference, height velocity and BMI. Biochemical parameters included peak growth hormone, serum IGF-1, IGFBP-3, acid labile subunit (ALS) and prolactin levels. Non-parametric testing was used to assess differences in growth and biochemical parameters.[br][bold]Results:[/bold] For the 66 children, mean age [plusmn] SD was 8.3 [plusmn] 3.3 years, with 65% being male. A total of 39 SNPs was identified in GH/IGF axis genes and 35 in primordial 3-M genes (59 patients had 1 SNP, 7 patients carried 2 or more SNPs [15 in total]). Growth parameters were as follows: Height SDS -2.9 (-6.0 to -2.0), weight SDS -0.8 (-3.7 to +1.1), height velocity 5.0 (1.1 to 10) cm/yr. Peak GH level was 17.9 [micro]g/L (range 7.0 to 58) and IGF-1 SDS was -1.9 (range -3.9 to +0.4). Parameters were then compared by each gene, by homozygotes versus heterozygotes, and by GH-IGF axis versus primordial 3-M SNPs: There were no significant differences between any of these groups.[br][bold]Conclusion:[/bold] It is not possible to define a growth or biochemical phenotype in ISS children associated with specific missense SNPs in growth disorder genes.[br][br]Nothing to Disclose: CB, MB, PD, LN, EP, PC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 752 198 1603 SUN-591 PO29-02 Sunday 1400 2012


1397 ENDO12L_SUN-592 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) A Novel Heterozygous [italic]OTX2[/italic] Deleterious Variant (p.H230L) in a Patient with Hypopituitarism and Ectopic Posterior Pituitary without Eye Malformation Michele Moreira, Marcela M Franca, Aline P Otto, Fernanda Correa, Ivo JP Arnhold, Berenice Bilharinho Mendonca, Sally Camper, Luciani R Carvalho Hospital das Clinicas, S[atilde]o Paulo, Brazil; University of Michigan, Ann Arbor, MI The incidence of short stature due to growth hormone (GH) deficiency occurs in 1:4,000-10,000 live births. Several transcription factors are necessary for the differentiation of five hormone producing cell types in the adenohypophysis. Patients with mutations in [italic]HESX1, GLI2, LHX3, LHX4, SOX2, SOX3, PROP1[/italic],and [italic]POU1F1[/italic] have been described in humans with pituitary hormone deficiencies. [italic]OTX2 [/italic]mutation can cause eye malformation such as anophthalmia and microphthalmia alone or in association with isolated GH deficiency (IGHD) or combined pituitary hormone deficiency (CPHD). Recently, two unrelated patients with CPHD and ectopic posterior pituitary lobe (EPP) without ocular abnormalities were found to harbor heterozygous [italic]OTX2[/italic] mutation suggesting a role for this gene in an etiology of hypopituitarim without other syndromic features.The aim of this study was to analyze [italic]OTX2 [/italic]in patients with IGHD or CPHD and correlate molecular findings with phenotype. We studied 142 Brasilian patients with CPHD (7 consanguineous parents and 33 relatives with short stature) and 44 with IGHD (7 consanguineous parents and 11 relatives with short stature). Pacients[apos] DNA samples were subjected to polymerase chain reaction using primers designed to amplify the translated exons and intron- exon borders. The PCR products were purified and sequenced by the Sanger method. A novel variant p.H230L in [italic]OTX2[/italic] was found in a single patient with CPHD associated with EPP without eye malformation. This variant was not found in 400 alleles from 200 Brasilian controls. The histidine at the position 230 is conserved across all vertebrate species and[italic] in silico[/italic] analysis predicts a deleterious effect of leucine substitution. Familial segregation reveled that the mother and two unaffected brothers are heterozygous carriers, suggesting incomplete penetrance. This idea is consistent with the observation that the features of mice heterozygous for [italic]Otx2[/italic] loss of function are strongly influenced by genetic background. We are assessing the function of this variant in cell culture assays and exploring the possibility of digenic inheritance with exome sequencing in the affected patient. In conclusion, our set of 186 patients with hormone deficiencies without ocular malformation is the largest population screened for mutations in [italic]OTX2.[/italic] The detection of only one suspicious variant in 186 individuals suggests that [italic]OTX2[/italic] is an uncommon cause of CPHD or IGHD without eyes malformation in the Brazilian population.[br][br]Nothing to Disclose: MM, MMF, APO, FC, IJPA, BBM, SC, LRC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 808 198 1604 SUN-592 PO29-02 Sunday 1401 2012


1398 ENDO12L_SUN-593 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Association of Polymorphisms in the VDR Promoter and ESR1 Intron 1 Region with Idiopathic Short Stature Kye Shik Shim, Seo Kyung Choi, Se Young Kim, Mun Sook Park, Chong-Woo Bae Kyung Hee University College of Medicine, Seoul, Korea; Bundang Jeseng General Hospital, Seongnam City, Korea [bold]Purpose:[/bold] The genetic alterations of vitamin D receptor (VDR) or estrogen receptor (ESR) are related with the growth of long bone, in other word, height. There were a lot of reports regarding association of polymorphisms in the VDR promoter and ESR1 intron 1 region with many disorders, but not with idiopathic short stature (ISS). We investigated the association of them with ISS.[br][bold]Method:[/bold] A total of 50 subjects, including 29 ISS patients and 21 healthy controls with their heights within the normal range were recruited. We selected two single nucleotide polymorphisms (SNPs) from VDR promoter (rs4516035 and rs11568820) and three from intron 1 of ESR1 (rs3778609, rs12665044 and rs827421) as candidates, respectively. The SNaPshot assay was performed according to the manufacturer[apos]s instructions (ABI PRISM SNaPShot Multiplex kit, Foster City, CA). Analysis was carried out using Genemapper ver. 4.0 (PE Applied Biosystems, Foster City, CA). The frequencies of allele and genotype, and the departures of the genotype distribution from Hardy-Weinberg equilibrium for each SNP were analyzed using the chi-square test or Fisher[apos]s exact test. Linkage disequilibrium was calculated with the Haploview ver.3.2. The genotype-specific risks were estimated as odds ratios with associated 95% confidence intervals using unconditional logistic regression analysis with SAS ver.8.02 (SAS Institute Inc., Cary, NC).[br][bold]Results:[/bold] In genotype analyses, the frequency of A/A genotype at the Cdx-2 binding site locus (rs11568820) upstream of exon 1e of VDR was decreased, but the one of G/G genotype at rs827421 of intron 1 of ESR1 was significantly increased in ISS patients.[br][bold]Conclusion:[/bold] The genetic variations at the Cdx-2 binding site of VDR promoter and ESR1 intron 1 region can be contributing factors of the growth of height.[br][br]Nothing to Disclose: KSS, SKC, SYK, MSP, C-WB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 887 198 1605 SUN-593 PO29-02 Sunday 1402 2012


1399 ENDO12L_SUN-594 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Factors Affecting Growth and Adult Height in Pediatric Renal Transplantation Minjae Kang, Haewoon Jung, Seung Joon Chung, Young Ah Lee, Choong Ho Shin, Sei Won Yang Seoul National University Children[apos]s Hospital, Seoul, Republic of Korea Background: Growth retardation is common in children with chronic kidney disease. Renal transplantation (TPL) resolves many endocrine, metabolic and uremic disturbances that contribute to growth retardation. However, growth continues to be suboptimal even after TPL. The aim of this study was to review factors affecting post-TPL growth and final adult height (FAH) in children who received kidney allografts.[br]Subjects and methods: A retrospective chart review of 65 patients (44 male, 21 female) who received renal TPL between 1983 and 2008 at Seoul National University hospital was done. Only patients less than 15 years of age at TPL with regular follow up for at least 3 years afterwards were included. Subjects with immediate graft failure or those receiving post-TPL growth hormone treatments were excluded. Height z-scores were recorded at diagnosis, commencement of renal replacement therapy (RRT), TPL and thereafter at 6 month intervals for 3 to 5 years. The delta height z-scores during 2 years and 5 years post TPL were calculated as the difference in height z-scores between those time points and height z-score at TPL.[br]Results: The mean age at TPL was 10.1 (1.8 [ndash] 14.4) years. The mean height z-score of recipients was -1.61[plusmn]1.36 (-4.64 [ndash] 1.64). The mean delta height z-score at 2 years and 5 years were 0.61[plusmn]0.89 (-1.03 [ndash] 2.41) and 0.38[plusmn]0.88 (-0.98 [ndash] 2.04) respectively. Age at TPL (before or after age 10) had a significant correlation with both 2 and 5 year change in height z-score ([italic]P[/italic]=0.001 and [italic]P[/italic]=0.012, respectively). Multivariate linear regression analysis showed that age and height at the time of TPL were significant determinants of 2 and 5 year growth after TPL (R[sup]2[/sup]=0.56, [italic]P[/italic]=0.006). Further analysis showed that height at commencement of RRT and duration of RRT were significant factors in determining the pre-TPL height (R[sup]2[/sup]=0.76, [italic]P[/italic][lt]0.001). Of 65 patients, 33 achieved FAH. The mean FAH was -1.21[plusmn]1.10 (-2.85 [ndash] 1.61) and the percentage of patients who attained a FAH z-score [ge]-1.88 was 76%. FAH z-score correlated with height z-score at the time of TPL ([italic]P[/italic][lt]0.001).[br]Conclusion: This study suggests that age and height at the time of TPL are important factors affecting post-TPL growth and FAH. Maximizing growth before TPL by decreasing the duration of RRT and early preemptive TPL may lead to better attainment of expected adult height.[br][br]Nothing to Disclose: MK, HJ, SJC, YAL, CHS, SWY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 503 198 1606 SUN-594 PO29-02 Sunday 1403 2012


1400 ENDO12L_SUN-595 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) A Prospective Study of Pubertal Growth in Children with Inflammatory Bowel Disease Avril Mason, Richard K Russell, Jonathan Bishop, Paraic McGrogan, S F Ahmed Royal Hospital for Sick Children Glasgow, Glasgow, UK; Royal Hospital for Sick Children Glasgow, Glasgow, UK [bold]Background[/bold]: Puberty is understood to be commonly affected in adolescents with Crohn[apos]s Disease (CD) and ulcerative colitis (UC). However, the extent of the related problems with growth, have rarely been quantified.[br][bold]Objective[/bold]: To determine the impact of CD and UC on pubertal growth.[br][bold]Methods:[/bold] Prospective study of 63 children with IBD:CD-M(23); CD-F(22); UC-M(12) and UC-F(6) with a median (range) age at diagnosis and age at baseline (T0) of 10.9; 10.9; 11.8; and 11.9yrs, and 13.4; 13.9; 13.4; and 13.2yrs, respectively.[br]Height at Diagnosis (HtAD) and height at T0(Ht0) and 12(Ht12) months was converted to SDS; and height velocity, between 0 and 12months converted to SDS adjusted for bone age (HVba) was calculated. Age at Tanner 1, Tanner 2/3 and Tanner 4/5 was compared to the normal population at 0 and 12months. C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and a PCDAI/PUCAI were compared, using 1 year cumulative data, with HV and HVSDS. Results were expressed as median (range).[br][bold]Results:[/bold][br]The median HT0SDS and HT12SDS was -0.15(-2.6;1.5); -0.32(-2.5;2.1); 0.27(-1.7;2.7); 0.18(-1.8;1.1) and -0.13(-2.5;1.6); 0.55(-3;2.1); 0.27(-1.6;2.2); 0.39(-1.3;0.6) respectively in CD-M, CD-F, UC-M and UC-F.[br]The median HVSDS was 0.1 (-2.3;3.6); -0.4 (-4;4.7);0.3 (-4.6;2.3); and 2.5 (-2;8.2) respectively in CD-M, CD-F, UC-M and UC-F. A statistically significant negative impact on parameter, Ht12SDS (p=0.0498) was seen in the CD-M group as compared to the normal population. Individually, 7/23 CD-M cases had one or more parameter affected: 6 subjects had HtADSDS [lt]-2, 3 subjects had Ht0SDS and Ht12SDS [lt]-2, and 1 subject had HVSDS [lt]-2. 3/22 CD-F cases had one or more parameter affected:1 subject had HtADSDS [lt]-2, 3 subjects had Ht0SDS and Ht12SDS [lt]-2, and 1 subject had HVSDS [lt]-2. In each of UC-F and UC-M 1 subject had HVSDS [lt]-2. No subjects remained Tanner 1 beyond the age at which 97% of population would be expected to be in Tanner 2. 1 CD-M case remained G2/3, at baseline, beyond the age at which 97% of male population would have expected to enter G4. 1 CDF case remained B2/3, at 12 months, beyond the age at which 97% of female population would have expected to enter B4. Median ESR showed a significant inverse association with HV in the whole group (r,-0.355; p=0.01) and the median PCDAI showed a significant inverse association with HVSDS in CD-M and CD-F(r[bold],[/bold]-0.40; p=0.02).[br][bold]Conclusion:[/bold] As a group, disorders of the pubertal growth spurt are more likely to occur in CD.[br][br]Nothing to Disclose: AM, RKR, JB, PM, SFA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1258 198 1607 SUN-595 PO29-02 Sunday 1404 2012


1401 ENDO12L_SUN-596 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Incidence of Pituitary Cysts in Patients with Growth Hormone Deficiency and Idiopathic Short Stature Joshua Rae, Jason Redman, Trisha Brown, Zhiyou Zhang, Melanie Rudnick, Michael Tenner, Richard A Noto New York Medical College, Valhalla, NY [underline]Introduction:[/underline] Cystic pituitary lesions are a common finding in autopsies, and when viewing CT and MRI scans. For the most part, they are considered a benign, incidental finding (1,2). However, they have been known to cause dysfunction including headaches, and endocrinopathies (3). The incidence of pars intermedia cysts (PICs) in the general population has been reported to be anywhere between 3% to 33% (4, 5). In this study, we investigated PICs in patients with growth hormone deficiency (GHD) and idiopathic short stature (ISS).[br][underline]Objective:[/underline] To determine and compare the incidence of pituitary cysts in GHD and ISS patients.[br][underline]Subjects:[/underline] 651 patients with a history of undergoing an MRI with particular attention to the pituitary gland.[br][underline]Methods:[/underline] A retrospective study of MRI imaging with contrast of the brain with particular attention to the pituitary gland was undertaken to determine incidence of cysts in all patients (AllPts). This group was further broken down into GHD patients and ISS patients. The incidence of cysts was calculated for all groups and compared with each other.[br][underline]Results[/underline]:[br]Mean age in the study group was 11.5 years +/-3.5). The cohort sex distribution was 35.6% female to 64.4% male. The incidence of cysts in the all patient group, GHD group and ISS group was 8.1%, 9.1% and 7.5% respectively. There was no difference in the incidence of cysts between these groups (p=.46). For all the patient group, 7.8% of females had cysts compared with 8.4% males. The incidence of cysts between male and females was not significant (P=7.9).[br]There was also no significant difference in the incidence of the GHD between males and females (p=.12). Lastly, there was no difference in the incidence of cysts in ISS group between males and females (p=.38)[br][underline]Discussion[/underline][br]The data presented here shows that the incidence of PICs in patients with GHD and ISS are no different from each other and from what has been found in the general population. They should probably be considered a benign condition. Preliminary follow-up of our patients, including patients on growth hormone therapy, has shown no major changes in the size of these cysts. However, long-term follow-up of these patients will be necessary to definitively determine their outcome.[br][br](1)Sanno N, Oyama K, Tahara S, Teramoto A, Kato Y. A survey of pituitary incidentaloma in Japan. Eur J Endocrinol. Aug 2003;149(2):123-127. (2)Takanashi J, Tada H, Barkovich AJ, Saeki N, Kohno Y. Pituitary cysts in childhood evaluated by MR imaging. AJNR Am J Neuroradiol. Sep 2005;26(8):2144-2147. (3)Baskin DS, Wilson CB. Transsphenoidal treatment of non-neoplastic intrasellar cysts. A report of 38 cases. J Neurosurg. Jan 1984;60(1):8-13. (4)Shanklin WM. On the presence of cysts in the human pituitary. Anat Rec. Aug 1949;104(4):379-407. (5) McGrath P. Cysts of sellar and pharyngeal hypophyses. Pathology. Apr 1971;3(2):123-131.[br][br]Nothing to Disclose: JR, JR, TB, ZZ, MR, MT, RAN 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1707 198 1608 SUN-596 PO29-02 Sunday 1405 2012


1402 ENDO12L_SUN-597 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Comparison of ICMA Versus RIA Measurements in Monitoring IGF-I Levels during rhGH Treatment in Short Children with Growth Hormone Deficiency (GHD), Turner Syndrome (TS), and Small for Gestational Age (SGA) Maria G Ballerini, Maria E Rodriguez, Debora Braslavsky, Horacio M Domene, Alicia Martinez, Ana Keselman, Paula Scaglia, Analia V Freire, Ignacio Bergada, Hector Jasper, Maria G Ropelato Hospital de Ni[ntilde]os Dr Ricardo Guti[eacute]rrez, Ciudad Aut[oacute]noma de Buenos Aires, Argentina [bold]Introduction:[/bold] Monitoring IGF-I levels during rhGH treatment is useful to ascertain efficacy, compliance and safety. To reach this purpose a reliable IGF-I assay is required. There is no data on the performance of ICMA IGF-I assays in children under rhGH therapy in comparison with classical extracted RIA. [bold]Aim:[/bold] To compare ICMA versus RIA assay[apos]s performance for measuring IGF-I in children under rhGH treatment. [bold]Subjects and Methods:[/bold] Serum IGF-I and IGFBP3 concentrations were measured in 13 GHD children (61 samples), 7 SGA children (30 samples) and 5 TS girls (16 samples), on rhGH (GHD dose: 0.16 mg/kg/week, SGA and TS: 0.33 mg/kg/week) for up to 3 years. IGF-I measurements: ICMA (IMMULITE 2000, Siemens), and an in house RIA (with prior alcohol-acid extraction followed by cryoprecipitation) (1). IGF-I levels were expressed in SDS according to reference values obtained in our laboratory. IGFBP-3 was measured by ICMA (IMMULITE 2000, Siemens). [bold]Results:[/bold] IGF-I ICMA values (ng/ml) were positively biased compared to RIA (ICMA =1.49 x RIA-13.7; r=0.97, p[lt]0.0001). [bold]Considering all samples [/bold]no relationship was found between the ratio IGF-I ICMA/RIA with neither the IGF-I RIA nor the IGFBP-3 levels; but a significant positive correlation was found for the SGA group between the IGF-I ICMA/RIA and IGFBP-3 levels (r=0.40, p=0.036). ICMA SDS results were systematically and significantly higher than RIA SDS (Mean [plusmn] SEM 1.20 [plusmn] 0.19 vs 0.57 [plusmn] 0.19, Wilcoxon test p[lt] 0.0001). Considering IGF-I + 2 SDS the cutoff for safety for patients on GH treatment, values above this limit were found in 13.1%, 36.7% and 68.8% by ICMA and in 4.9%, 26.7% and 56.3% by RIA of GHD, SGA, and TS samples respectively. The difference in proportions between groups was not significant (p[gt]0.05). In 12/107 (11%) samples, ICMA gave IGF-I levels above the safety cutoff while RIA results were below that limit. [bold]Conclusions:[/bold] ICMA assay overestimated IGF-I concentration in comparison with an extractive RIA in children under GH therapy. The positive relationship between the IGF-I ICMA/RIA ratio and IGFBP-3 concentration in SGA on high dose rhGH treatment suggests that ICMA procedure is not totally effective to circumvent IGFBPs interference. Both assays showed similar proportion of samples with IGF-I levels higher than 2SDS. It should be noted that ICMA, even when the results are expressed in SDS, would lead to a misinterpretation of IGF-I results, giving falsely elevated values in 11% of the samples.[br][br](1) Martinez AS et al, J Clin Endocrinol Metab 2000;85:4168.[br][br]Sources of Research Support: Grants from the Agencia Nacional de Promoci[oacute]n Cient[iacute]fica y Tecnol[oacute]gica (BID 1201/OC-AR PICT-2003 Nro. 05-14354) and from Consejo de Investigaci[oacute]n en Salud, Ministerio de Salud, Gobierno de la Ciudad Aut[oacute]noma de Buenos Aires.[br][br]Nothing to Disclose: MGB, MER, DB, HMD, AM, AK, PS, AVF, IB, HJ, MGR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 237 198 1609 SUN-597 PO29-02 Sunday 1406 2012


1403 ENDO12L_SUN-598 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Pharmacological Stimulated Growth Hormone Cut-Off Values Using an ICMA Assay Calibrated with an International Recombinant Human GH Standard 98/574 Juan Manuel Lazzati, Eduardo Chaler, Maria Gabriela Ballerini, Gabriela Ropelato, Mercedes Maceiras, M Frusti, Ignacio Bergada, Marco A Rivarola, Alicia Belgorosky Hospital de Pediatria Garrahan, Buenos Aires, Argentina; Hospital de Ni[ntilde]os [quot]Ricardo Gutierrez[quot], Buenos Aires, Argentina [bold]Background[/bold][bold]:[/bold] Different cut-off values, from 3 to10 ng/ml, depending on the assays, have been defined for pharmacological test (PhT) of growth hormone (GH) secretion, based on biochemical and clinical studies. Recently, a 22K recombinant GH isoform IRP IS 98/574 have been commercialized. We had previously defined a 6.1 ng/ml value, in terms of IMM IRP IS 80/505 as serum GH cut-off limit in PhT of GH secretion. Our aim was to assess which is the GH deficiency (GHD) diagnostic cut-off limit of serum GH in PhT of GH secretion for IMM immunoassay with the IRP IS 98/574.[br][bold]Methods: [/bold]We analyzed serum GH concentration in 138 serum samples, using IMM calibrated with both IS 80/505 and IS 98/574. Blood samples (n=138), from 92 different individuals, (103 males and 35 female; age range, 2.5 [ndash] 15.0 years) were used. All samples were selected from arginine and clonidine PhT.[br][bold]Results[/bold][bold]: [/bold]We found high significant linearity (y = 0.7307x + 0.2768, p[lt]0.001) between IMM IS 98/574 (y) and IMM IS 80/505 (x). The y value for fixed x value of 6.1 ng/ml in the IMM IS 80/505 was calculated, and a diagnostic cut-off limit for IMM IS 98/574 was estimated as 4.7 ng/ml. IMM IS 98/574/IMM IS 80/505 ratio plot and 95% prediction interval was 0.79 [plusmn] 0.17. Linearity and Bias were reconfirmed by Passing Bablock analyses and the Wilcoxon test respectively.[br][bold]Conclusions[/bold][bold]: [/bold]High significant linearity and differences among the assay with different IS were found. To suspect the diagnosis of GHD, we propose 4.7 ng/ml as the serum GH cut-off limit value in PhT for IMM immunoassay with the IRP IS 98/574as. Finally for each GH assay, an appropriate cut-off value for serum GH maximum peak response to PhT should be strongly recommended.[br][br]Sources of Research Support: FONCYT,CONICET, Argentina.[br][br]Nothing to Disclose: JML, EC, MGB, GR, MM, MF, IB, MAR, AB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 335 198 1610 SUN-598 PO29-02 Sunday 1407 2012


1404 ENDO12L_SUN-599 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Early Start of Growth Hormone (GH) Treatment Leads to Significantly Better Growth Outcome in GH Deficient Children When Assessed at near Adult Height: Data from the NordiNet[reg] International Outcome Study Lars Savendahl, Oliver Blankenstein, Michael Schlumpf, Birgitte Tonnes Pedersen, Isabelle Oliver Karolinska Institutet, Stockholm, Sweden; Charit[eacute]-Universit[auml]tsmedizin, Berlin, Germany; Novo Nordisk Health Care AG, Zurich, Switzerland; Novo Nordisk A/S, S[oslash]borg, Denmark; H[ocirc]pital des Enfants, Toulouse, France [bold]Introduction[/bold][br]Improving adult height is the major goal of growth hormone (GH) treatment in short children with growth hormone deficiency (GHD). Limited data are available on the impact of age at treatment start on adult height.[br][bold]Objective[/bold][br]To investigate the influence of age at treatment start on adult height in short GHD children.[br][bold]Methods[/bold][br]From the NordiNet[sup][reg][/sup] International Outcome Study (IOS) database, GHD children treated with Norditropin[sup][reg][/sup] were identified. Only patients who had reached near adult height (NAH) were included. NAH was defined as height at last visit when:[br]- age [ge]18 years [italic]or[/italic][br]- boys were [ge]16 years and height velocity (HV) [lt]2cm/year [italic]or[/italic][br]- girls were [ge]15 years and HV [lt]2cm/year[br]Included patients were divided into two groups based on age at treatment start:[br]- Group A: Early start of GH treatment: girls: age [lt]8 years; boys: age [lt]9 years[br]- Group B: Late start of GH treatment: girls: age [ge]8 years; boys: age [ge]9 years[br]Results are presented as mean [plusmn] SD.[br][bold]Results[/bold][br]In total 154 children were included, 34 in group A and 120 in group B. Mean age at treatment start was for group A 5.8[plusmn]2.4 years (47.1% girls) and for group B 11.7[plusmn]1.8 years (34.2% girls). Height SDS at treatment start was lower in group A than in group B (-3.4[plusmn]1.4 SDS [italic]vs.[/italic] -2.9[plusmn]1.2 SDS; p[lt]0.05[italic])[/italic] and the duration of GH treatment was longer in group A than in group B (10.4[plusmn]2.2 vs. 5.6[plusmn]1.2 years; p[lt]0.0001). No differences between groups A and B were found for average relative GH dose (32.8[plusmn]9.3 [italic]vs.[/italic] 31.4[plusmn]11.1 [micro]g/kg/day, respectively) and midparental height SDS (-0.57[plusmn]1.01 [italic]vs.[/italic] -0.73[plusmn]1.02 SDS, respectively). Patients in group A had a significantly better growth outcome as determined by height SDS increment from treatment start to NAH (2.55[plusmn]1.39 vs. 1.46[plusmn]1.18 SDS; p[lt]0.0001). Despite being significantly shorter at treatment start, patients in group A tended to be taller at NAH than patients in group B (-0.95[plusmn]1.36 [italic]vs.[/italic] -1.45[plusmn]1.49 SDS; p=0.081).[br][bold]Conclusion[br][/bold]Early start of GH treatment leads to significantly better long-term growth outcome in GH deficient children.[br][br]Disclosures: LS: Committee Member, Novo Nordisk. OB: Committee Member, Novo Nordisk. MS: Employee, Novo Nordisk. BTP: Employee, Novo Nordisk. IO: Committee Member, Novo Nordisk. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 489 198 1611 SUN-599 PO29-02 Sunday 1408 2012


1405 ENDO12L_SUN-600 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Early Markers of Cardiovascular Risk in Children with Growth Hormone (GH) Deficiency (GHD): Effects of 2 Years of Replacement Therapy Donatella Capalbo, Lucia De Martino, Andrea Esposito, Iolanda Di Donato, Flavia Barbieri, Giuseppina Mattace Raso, Rosaria Meli, Mariacarolina Salerno University Federico II, Naples, Italy; University Federico II, Naples, Italy [bold]Background:[/bold] Adults with Growth Hormone (GH) deficiency (GHD) may have an increased risk of cardiovascular disease. Several studies showed that also GHD children may present cardiovascular risk factors. However results are controversial.[br][bold]Objective:[/bold] Aim of this prospective, case-control study on a large cohort of children with GHD was to evaluate the effects of GHD and GH therapy on cardiovascular risk factors, such as lipid profile, clinical measures of visceral adiposity and inflammatory markers in a large cohort of GHD children.[br][bold]Subjects and Methods:[/bold] Total-, LDL- and HDL- cholesterol, triglycerides, fibrinogen, C reactive protein (CRP), waist circumference, waist-to-height (WHtR) and waist-to-hip ratio (WHR) were evaluated in 60 GHD children (40 males), aged 9.9[plusmn]0.4 years, before and after 2 years of GH therapy. 60 healthy, age-, sex- and BMI-matched healthy controls were enrolled.[br][bold]Results: [/bold]Compared with controls, GHD children at baseline had higher total-cholesterol (163.8[plusmn]3.0vs150[plusmn]2.8 mg/dl, p=0.001), LDL-cholesterol (96.7[plusmn]2.7vs83.3[plusmn]3.7 mg/dl, p=0.003), triglycerides (73.2[plusmn]5.4vs59.5[plusmn]3.6 mg/dl, p[lt]0.04), fibrinogen (305[plusmn]8.4vs278[plusmn]9 mg/dl, p=0.03) and CRP (0.4[plusmn]0.03vs0.31[plusmn]0.006, p=0.001).Waist circumference was slightly higher in GHD patients although this difference was not significant. However, GHD children had higher WHtR (0.54[plusmn]0.01vs0.47[plusmn]0.01, p[lt]0.0001) and WHR (0.96[plusmn]0.01vs0.92[plusmn]0.01, p=0.005) than controls. Two years of GH therapy were associated with significantly reduced levels of total- (149.4[plusmn]3.27, p=0.001) and LDL-cholesterol (79.6[plusmn]3.0, p[lt]0.0001), triglycerides (60.9[plusmn]3, p[lt]0.04), fibrinogen (265[plusmn]6.5, p=0.0003), CRP (0.32[plusmn]0.01, p=0.002), WHtR (0.49[plusmn]0.01, p=0.0006) and WHR (0.92[plusmn]0.01, p[lt]0.005).[br][bold]Conclusions: [/bold]Our data suggest that in children GHD is associated with cardiovascular risk factors such as adverse lipid and inflammatory profile and increased visceral adiposity. GH replacement therapy improves body composition, lipid profile and it reduces inflammatory risk factors.[br][br]Nothing to Disclose: DC, LDM, AE, IDD, FB, GMR, RM, MS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 923 198 1612 SUN-600 PO29-02 Sunday 1409 2012


1406 ENDO12L_SUN-601 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Race, Ethnicity, Gender, Age and Height Differences in Children Treated with Recombinant Human Growth Hormone (rhGH): Have They Changed over Time? Data from the Genentech National Cooperative Growth Study (NCGS) Joseph Cernich, James W Frane, Barbara Lippe Children[apos]s Mercy Hospitals and Clinics, Kansas City, MO; , Santa Monica, CA; Genentech, Inc, South San Francisco, CA Background: The first published analysis of NCGS data for 2300 patients from the period 1986-1988 indicated that Caucasian (C) males (M) were disproportionally represented and relatively taller using height standard deviation scores (HSDS) compared to C females (F), African-American (AA) M and AAF [1].[br]Hypothesis: That as rhGH became more accessible there might be a shift in treatment (Rx) to a more balanced gender, racial, age and ethnic profile.[br]Methods: For this report we focused on prepubertal children at entry (to be able to assess HSDS differences) and racial disparity among AA and Asians. Hispanics were initially categorized as [ldquo]others[rdquo] and only later were separate, so Others includes them. 40,006 patients, naive to prior GH Rx, prepubertal and at most 14 yr at baseline were assessed. Patients were divided into 3 date categories: [1] prior to 1/1990, [2] 1/1990-12/1999, and [3] 1/2000 to 12/2009.[br]Results: The proportion of new AA patients were [1] 4.8%;[2] 5.1%, and [3] 3.7% while the % AA children in the general population was 14-16% during these times. The proportion of C fell from 86.2% to 82.5% to 79.1% while the Other category rose from 6.4 to 9.9 to 14.9%. The Asian category stayed at [sim]2.5%. The proportion of F was [sim]37% overall and similar for C and AA females and slightly higher for Asians (41%) and Others (46%).[br]AA M were younger than the CM and in both mean age increased by the last time interval: [1] 7.3 vs 8.4 yrs [2] 7.3 vs 8.6,[3] 7.6 vs 9.1 [italic]but [/italic]AAM had shorter SDS scores although in both groups they tended to increase over time: [1] -3.2 vs -2.8, [2]-3.0 vs -2.6 [3]-2.7 vs-2.4. Similarly AA F were younger and shorter than CF with some increase in age over time. The Other category was similar to the AA group.[br]Discussion: The disparity between the representation of C vs AA children by relative number and the disparity between M vs F in all groups has not changed over time. Similarly the fact that AA children are younger and shorter when diagnosed, as were the Others, coupled with the low proportionate representation of AA and the emergence of the Others, suggests that the racial and ethnic disparity results in only the most severely affected of such children being treated. Conclusion: Access to or recognition of the need for rhGH treatment has not changed over time and remains unbalanced with greater use in the C population and use otherwise limited to the more severely affected children in AA and Other groups.[br][br](1) August GP et al., J Pediatr 1990;116:899.[br][br]Sources of Research Support: This study was supported by Genentech,Inc.[br][br]Disclosures: JC: Speaker Bureau Member, Pfizer, Inc., Teva. JWF: Consultant, Genentech, Inc. Ipsen. BL: Consultant, Genentech, Inc.; Employee, Genentech, Inc. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 664 198 1613 SUN-601 PO29-02 Sunday 1410 2012


1407 ENDO12L_SUN-602 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Evaluation of Growth Response in Short Prepubertal Children Suffering from a Disease with Growth Retardation and Treated with Somatropin: A Prospective, Longitudinal Non-Randomized, Open, Phase II Multicenter Study in Germany Helmuth-Guenther Doerr University of Erlangen, Erlangen, Germany Idiopathic short stature (ISS) is a common reason for referral to the pediatric endocrine clinic but therapeutic options for these children are limited. We designed a study to include patients who suffer from growth retardation caused by a non-registered indication for Somatropin treatment. [bold]Objective:[/bold] The primary objective was to determine whether GH therapy showed an improvement of height and change in height (SDS) after one year. [bold]Methods:[/bold] After 12 months of GH therapy, the height velocity SDS (HV-SDS) should increase [gt] 1 SD, and/or delta height [gt] 0.7 SD. The study duration was one year and continuation in the study after one year for the second year was possible after an interim analysis of the growth data and if funding by health insurance was rejected. [bold]Patients:[/bold] We studied 125 prepubertal children (76 m, 49 f) from 13 pediatric centres in Germany who fulfilled the inclusion criteria: severe growth retardation ([lt] -2,5 height SDS), annual HV-SDS [lt] 0 SD, parental adjusted target height [lt] -1 SD; CA [gt] 4 and [lt] 10 years, girls: Tanner B1, boys: testis [le] 3 ml; any disease which is not part of the registered indications for GH treatment in Germany. The GH dose was 0.035 mg/kg/d (s.c. injections at bedtime). [bold]Results:[/bold] We identified three main groups: Idiopathic short stature, ISS (n=67), familial short stature, FSS (n=38), Noonan syndrome, NS (n=12). These three diagnoses covered 95 % of all reported diagnoses of the subjects. Overall, 123 subjects completed one year of GH therapy. After start of GH therapy HV-SDS increased significantly between start and month 12 in children with ISS, FSS and NS (ISS: p[lt]0.0001; FSS: p[lt]0.0001; Noonan: p=0.001). The mean ([plusmn] SD) HV-SDS increase was 5.27 ([plusmn]2.33) in ISS, 4.66 ([plusmn]2.04) in FSS, and 3.36 ([plusmn]1.58) in NS. The differences between the groups were statistically significant (p=0.015). Pair wise tests showed significant differences between subjects with ISS vs. NS (p=0.003), between FSS vs. NS (p=0.0424), but not between ISS vs. FSS (p=0.287). Mean height SDS improved significantly from -3.50 ([plusmn]0.79) at baseline to -2.72 ([plusmn]0.89) at month 12 (p[lt]0.0001). With regard to the three groups, the mean increase in HSDS was highest in ISS (0.83 [plusmn] 0.27), followed by FSS (0.79 [plusmn] 0.24), and lowest in NS (0.60 [plusmn] 0.23). [bold]Conclusions:[/bold] In accordance with the protocol after one year of GH treatment, 121 (98.4%) subjects qualified as responders while only two subjects (1.63%) did not. Both non-responders dropped out of the study.[br][br]Nothing to Disclose: H-GD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 731 198 1614 SUN-602 PO29-02 Sunday 1411 2012


1408 ENDO12L_SUN-603 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) An Analysis of the Age-Based Relationship between per-m[sup]2[/sup] and per-kg Dosing for Growth Hormone from the OZGROW Database: Implications for Maximizing GH Efficacy Ian Paul Hughes, Catherine Choong, Mark Harris, Andrew Cotterill Mater Hospital, South Brisbane, Australia; Princess Margaret Hospital for Children, Subiaco, Australia [bold]Introduction[/bold][br]In Australia growth hormone (GH) for children is prescribed by mg/m[sup]2[/sup]/wk of body surface area (BSA). In most other countries paediatric GH-doses are given in as mg/kg/wk or an equivalent, eg. [micro]g/kg/day. There is no direct conversion between the two formats as the/m[sup]2[/sup] protocol requires both the weight and height of the patient for its calculation. BSA is estimated by sqrt((Height(cm)XWeight(kg))/3600). This makes interpretation of doses given in the literature problematic. A second question that arises relates to the relative efficacy of each dosing format. To better understand the relationship between formats and the possible implications to treatment efficacy we analysed GH doses from the OZGROW database with respect to the age of the patient.[br][bold]Subjects and Methods[/bold][br]10,092 doses from 570 Turner syndrome (TS), 5358 doses from 239 GH deficient (GHD), 1966 doses from 165 Prader-Willi syndrome (PWS) patients and 12,374 doses from 730 patients under the indication of [ldquo]Short Stature and Slow Growth[rdquo] (SSSG) were extracted. All doses were prescribed as mg/m[sup]2[/sup]/wk and each was converted to mg/kg/wk. Initially, all doses, for each format and diagnosis, were plotted against age and the relationship summarized by a linear regression (LR) equation. Subsequently, common dose ranges (in mg/m[sup]2[/sup]/week) were plotted in both formats.[br][bold]Results and Discussion[/bold][br]Plotting the/m[sup]2[/sup] doses revealed a slight increase in dose with age for each diagnosis with the slope (mg/m[sup]2[/sup]/wk/year) greatest for TS (0.23), then SSSG (0.13), GHD (0.12), and PWS (0.02). These results reflect the policy of incremental dosing (ID) for which there is greatest scope in TS. However, the same doses in terms of/kg decreased with age. Slopes (mg/kg/wk/year) were TS (-0.0010), SSSG (-0.0015), GHD (-0.0019), and PWS (-0.0066). This was accentuated when specific dose ranges were used reducing the ID effect. For 4.0-5.5mg/m[sup]2[/sup]/wk doses in SSSG the slope of the/kg doses was -0.0052. For TS, these slopes decreased to -0.0067 (4.0-5.5), -0.0099 (6.0-7.5), and -0.0093 (8.0-9.5). The decrease in equivalent/kg doses appeared linear but may be exponential for the first year of life.[br][bold]Conclusions[/bold][br]Per m[sup]2[/sup] dosing results in relatively high/kg doses in young patients that decrease with age. The lowest prescribed dose of 4.5mg/m[sup]2[/sup]/wk is equivalent to approx. 0.22mg/kg/wk at 6months but only 0.12mg/kg/wk at 16years. A doseXage interaction for GH response reported for/m[sup]2[/sup] dosing may be explained by this observation.[br][br]Sources of Research Support: Australasian Paediatric Endocrine Group.[br][br]Nothing to Disclose: IPH, CC, MH, AC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1618 198 1615 SUN-603 PO29-02 Sunday 1412 2012


1409 ENDO12L_SUN-604 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Quantifying Adherence to Growth Hormone Treatment: The Easypod[trade] Connect Observational Study (ECOS) Peter SW Davies, Martin Borkenstein, Evangelia Charmandari, Ho-Seong Kim, Jeremy Kirk, Ludmila Kostalova, Jan Lebl, Sandro Loche, Andrea Luczay, Marc Nicolino, Svante Norgren, Dolores Rodriguez Arnao, John Vandermeulen, Christoph Gasteyger, Jurgen Zieschang, Monia Zignani University of Queensland, Brisbane, Australia; Medical University Graz, Graz, Austria; Aghia Sophia Children[apos]s Hospital, Athens, Greece; Yonsei University, Seoul, Korea; Birmingham Children[apos]s Hospital, Birmingham, UK; Comenius University Medical School, Bratislava, Slovakia (Slovak Republic); Charles University in Prague, Prague, Czech Republic; Ospedale Microcitemico - ASL Cagliari, Cagliari, Italy; Semmelweis University, Budapest, Hungary; Lyon University, Lyon, France; Karolinska University Hospital, Stockholm, Sweden; Hospital Universitario Gregorio Mara[ntilde][oacute]n, Madrid, Spain; McMaster Children[apos]s Hospital and McMaster University, Ontario, Canada; Merck Serono SA, Geneva, Switzerland; Merck KGaA, Darmstadt, Germany BACKGROUND: Recombinant human growth hormone (r-hGH) is indicated for pediatric patients with a variety of growth disorders. Until recently, analysis of adherence to r-hGH treatment, which may be required for many years, has been limited by recall bias and reliance on self-reporting. Accurate recorded data on r-hGH use can now be collected in patients using the easypod[sup]TM[/sup] auto-injector. The multinational easypod[sup]TM[/sup] connect observational study (ECOS) was launched in 2010 to collect and analyze r-hGH dosing and clinical and auxological data from patients prescribed r-hGH via easypod[sup]TM[/sup]. Nine countries are currently recruiting in ECOS.[br]OBJECTIVES: The primary objective is to assess adherence in patients receiving r-hGH via easypod[sup]TM[/sup]. Secondary objectives include describing the impact of adherence on clinical outcomes for patients receiving r-hGH and identifying adherence patterns.[br]DESIGN/METHODS: Data will be obtained from patients[apos] medical notes and uploaded from the auto-injectors. Auxological parameters (height, weight, Tanner puberty stage, bone age) are collected and prescribed dosing data recorded at regular clinic visits, up to three times yearly as per routine clinical practice. Adherence at 1 year will be calculated (number of days the patient administered injections divided by the expected number of injection days over 1 year [365 days], expressed as a percentage). Dose intensity (defined as total amount of dose received divided by planned amount of dose over 1 year) will be analyzed. Data from auto-injectors will provide a complete record of a patient[apos]s r-hGH use. Adherence data will then be correlated with clinical outcomes. An adherence pattern will also be developed based on patients[apos] age, sex, indication, self-injection, and time on treatment. The study will run until 2015, with yearly analyses. An external scientific steering committee will oversee the study.[br]CONCLUSIONS: With data from ECOS, for the first time it will be possible to accurately assess r-hGH treatment adherence in various growth disorders and explore its potential impact on growth outcomes. Ultimately, drivers of and barriers to r-hGH treatment adherence will be identified, allowing appropriate support programs to be developed.[br][br]Sources of Research Support: Merck KGaA (Australia, Czech Republic, France, Greece, Hungary, Italy, Slovakia, Sweden, UK), all affiliates of Merck KGaA, Darmstadt, Germany.[br][br]Disclosures: PSWD: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A. MB: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A. EC: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A. H-SK: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A. JK: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A.; Speaker, Merck Serono S.A.; Researcher, Merck Serono S.A. LK: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A. JL: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A. SL: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A. AL: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A. MN: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A. SN: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A. DRA: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A. JV: Planning Group Member, Merck Serono S.A.; Investigator, Merck Serono S.A. CG: Employee, Merck Serono S.A. JZ: Employee, Merck Serono S.A. MZ: Employee, Merck Serono S.A. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 606 198 1616 SUN-604 PO29-02 Sunday 1413 2012


1410 ENDO12L_SUN-605 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Diagnosis of Growth Hormone Deficiency in the Transition Period Natascia Di Iorgi, Marco Cappa, Lucia Ghizzoni, Sandro Loche, Giorgio Radetti, Maria Carolina Salerno, Donatella Capaldo, Bianca Ofelia Iovovich, Costanza Frassinetti, Serena Noli, Mohamad Maghnie IRCCS G Gaslini, University of Genova, Genova, Italy; IRCCS Ospedale Pediatrico Bambin Ges[ugrave], Roma, Italy; Azienda Ospedaliera Universitaria San Giovanni Battista-Molinette, Torino, Italy; PO Microcitemico, ASL Cagliari, Cagliari, Italy; Ospedale Regionale di Bolzano, Bolzano, Italy; Universit[agrave] di Napoli Federico II, Napoli, Italy; IRCCS G Gaslini, University of Genova, Genova, Italy [bold]Objectives.[/bold] To reassess GH response in young adults with childhood-onset GHD (COGHD).[br][bold]Methods.[/bold] We present the preliminary data of 69 subjects (30F, 39M) recruited from a multicenter cross-sectional observational study, in whom anthropometrics, ITT (n=53), GHRH-arginine (n=67), IGF-1 evaluations were undertaken at a mean age of 17.3[plusmn]1.8yrs. Thirty-three had idiopathic GHD (iGHD), 36 secondary GHD (SGHD) due to brain tumors (n=35) or LMA (n=1). Isolated GHD (IGHD) was found in 45 (n=29 iGHD, n=16 SGHD) and MPHD in 23 (n=4 iGHD, n=19 SGHD). Peak GH values [gt]6[mu]g/L for ITT and [gt]19[mu]g/L for GHRH-arginine were considered normal (1,2).[br][bold]Results. [/bold]Peak responses to ITT (P=0.002) and GHRH-arginine (P=0.0003) were significantly lower in SGHD (5,2[plusmn]6,0 and 14,2[plusmn]12,0[mu]g/L, respectively) compared to iGHD subjects (13,0[plusmn]11,6 and 19,8[plusmn]14,4[mu]g/L, respectively); there were no differences in mean IGF-1 values (-1,9[plusmn]1,9 and -1,2[plusmn]1,0 SDS, respectively). Peak responses to ITT and to GHRH-arginine were significantly lower in SGHD whit IGHD compared to iGHD with IGHD (P=0.0039 and P=0.03, respectively), while patients with MPHD did not display any differences in GH peak responses after both stimulation test. Subjects with SGHD and IGHD showed no differences in GH peak after ITT compared to those with MPHD; in contrast patients with SGHD and IGHD showed significantly higher GH responses after GHRH-arginine compared to those with MPHD (P[lt]0,005). In iGHD IGF-1 values were significantly lower in MPHD compared to IGHD (-2,5[plusmn]0,3 and -1,1[plusmn]0,9 SDS, P=0.02), while there was no significant differences in SGHD subjects whether they had IGHD or MPHD (-1,6[plusmn]1,8 and -2,1[plusmn]1,9 SDS).[br]A GH peak response[bold] [lt][/bold]6[mu]g/L for ITT was found in 28/53 of whom 22 SGHD (n=14 MPHD, n=8 IGHD) and 6 iGHD (n=3 MPHD, n=3 IGHD); a GH peak [lt]19[mu]g/L for GHRH-arginine was obtained in 35/67 of whom 26 SGHD (n=17 MPHD, n=9 IGHD) and 9 IGHD (n=3 MPHD, n=6 IGHD). Mean BMI SDS was significantly higher in patients with SGHD compared to iGHD (1,3[plusmn]2.2 vs 0.0[plusmn]1,5; P=0.01). A negative correlation was found between BMI SDS and GH peak to ITT (r=-0.56, P=0,01 for iGHD, r=-0.36, P=0.07 for SGHD) or GH peak to GHRH-arginine (r=-0.58, P=0,0008 for iGHD, r=-0.44, P=0.01 for SGHD).[br][bold]Conclusions[/bold]: Patients with childhood onset SGHD are at higher risk of permanent GHD compared to iGHD. ITT confirms to be reliable in the identification of patients who may need rhGH treatment in adult life. BMI affects more profoundly GH response after GHRH-arginine.[br][br](1) Ho KK; 2007 GH Deficiency Consensus Workshop Participants, Eur J of Endocrinol 2007;157:695. (2) Ginevra Corneli et al., Eur J of Endocrinol 2007;157:701.[br][br]Sources of Research Support: A Grant from Pfizer Care.[br][br]Nothing to Disclose: NDI, MC, LG, SL, GR, MCS, DC, BOI, CF, SN, MM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 754 198 1617 SUN-605 PO29-02 Sunday 1414 2012


1411 ENDO12L_SUN-606 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Attaining Target Height: Outcomes in Children Treated with Growth Hormone for Isolated Growth Hormone Deficiency, Multiple Pituitary Hormone Deficiency, Children Born Small for Gestational Age, Idiopathic Short Stature, and Chronic Renal Insufficiency [mdash] Analysis of Data from the ANSWER Program[reg] Judith L Ross, Peter A Lee, Robert Z Gut, Vatsala Karwe, John A Germak Thomas Jefferson University, DuPont Hospital for Children, Wilmington, DE, Philadelphia, PA; Pennsylvania State College of Medicine, The Milton S Hershey Medical Center, Hershey, PA; Novo Nordisk, Inc, Princeton, NJ [bold]Background and Objective:[/bold] Long-term safety and efficacy data on pediatric patients treated with Norditropin[sup][reg] [/sup](somatropin rDNA origin) are being collected through the American Norditropin Studies: Web-Enabled Research (ANSWER) Program[sup][reg][/sup], a US-based registry. Height outcomes were assessed in patients with isolated growth hormone deficiency (IGHD), multiple pituitary hormone deficiency (MPHD), idiopathic short stature (ISS), small for gestational age (SGA), and chronic renal insufficiency (CRI).[br][bold]Methods:[/bold] By October 2011, data were collected from GH-naive children with IGHD (n=5024), MPHD (n=436), SGA (n=647), ISS (n=1096), and CRI (n=49). Change from baseline in height standard deviation scores ([Delta]HSDS) and corrected HSDS (calculated as the difference between HSDS and target HSDS) were analyzed over 5 years. Three years of data were included for CRI subjects because of a small patient sample.[br][bold]Results:[/bold] At baseline, patients with MPHD (7.3[plusmn]5.4 y), SGA (8.3[plusmn]3.9 y), and CRI (8.1[plusmn]4.8 y) were generally younger than patients in other diagnostic categories (10.9[plusmn]3.5 y for IGHD and 11.2[plusmn]3.1 y for ISS). The lowest mean peak GH levels at baseline were observed in MPHD (3.6[plusmn]3.5 ng/mL) and IGHD (5.9[plusmn]2.6 ng/mL) groups, vs other indications (CRI, 12.5[plusmn]11.0 ng/mL; SGA, 14.6[plusmn]9.7 ng/mL; ISS, 17.1[plusmn]9.1 ng/mL). Patients with MPHD reached the greatest mean HSDS (baseline, [minus]1.9[plusmn]1.4; y5, [minus]0.4[plusmn]1.3) compared to other indications after 5 years of treatment. Patients with IGHD, MPHD, SGA, and ISS who completed 5 years of treatment showed increases in [Delta]HSDS (IGHD: y1, 0.6[plusmn]0.5; y5, 1.7[plusmn]0.9; MPHD: y1, 0.6[plusmn]0.9; y5, 1.6[plusmn]1.3; SGA: y1, 0.6[plusmn]0.5; y5, 1.7[plusmn]0.8; ISS: y1, 0.5[plusmn]0.4; y5, 1.6[plusmn]0.9); patients with CRI also showed an increase after 3 years (y1, 0.3[plusmn]0.6; y3, 0.5[plusmn]1.2). At y5, the corrected HSDS improved considerably in patients with ISS (baseline, [minus]1.5[plusmn]1.0; y5, [minus]0.2[plusmn]0.7), MPHD (baseline, [minus]1.9[plusmn]1.4; y5, -0.2[plusmn]1.3), IGHD (baseline, [minus]1.7[plusmn]1.1; y5, [minus]0.3[plusmn]0.9), and SGA (baseline, [minus]1.9[plusmn]1.2; y5, [minus]0.7[plusmn]1.0). CRI subjects showed improvement in corrected HSDS at y3 (baseline, [minus]2.2[plusmn]1.7; y3, [minus]0.6[plusmn]1.5).[br][bold]Conclusions:[/bold] Overall, the results indicate that GH treatment increased [Delta]HSDS, and corrected HSDS for IGHD, MPHD, ISS, and SGA groups at year 5, and for CRI subjects at year 3. GH treatment was associated with HSDS approaching genetic height potential, based on target heights, with greater corrected HSDS observed in children with MPHD, IGHD, and ISS compared with CRI and SGA patients.[br][br]Disclosures: JLR: Advisory Group Member, Novo Nordisk, Eli Lilly & Company, Abbott Laboratories; Clinical Researcher, Novo Nordisk, Pfizer, Inc., Eli Lilly & Company. PAL: Advisory Group Member, Abbott Laboratories, Novo Nordisk; Clinical Researcher, Abbott Laboratories, Eli Lilly & Company, Pfizer, Inc., Ipsen. RZG: Employee, Novo Nordisk. VK: Employee, Novo Nordisk. JAG: Employee, Novo Nordisk. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2257 198 1618 SUN-606 PO29-02 Sunday 1415 2012


1412 ENDO12L_SUN-607 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Norditropin[reg] Pen Users May Experience Less Pain [amp] Stinging upon Injection Compared with Other rhGH Products Victoria Divino, Tin-Ming Douglas, John Germak, Smith Chioma, Petrilla Allison, Lee Chan Won, Wisniewski Tami IMS Consulting Group, Falls Church, VA; Novo Nordisk Inc, Princeton, NJ; Novo Nordisk Inc, Princeton, NJ [bold]Purpose: [/bold]Daily injections of rhGH may add to the therapeutic burden of a child. However, the perception of pain and stinging associated with rhGH products is not well understood.[br][bold]Methods:[/bold] A post-hoc analysis of a survey, conducted in partnership with the MAGIC Foundation, evaluated the perception of injection-related pain/stinging in children receiving rhGH therapy. Caregivers were required to respond as proxies for patients [lt]13 years, but could respond for patients of any age. Bivariate analyses were performed to examine demographic differences among rhGH users. Multivariable analyses were performed to identify factors associated with the perception of pain/stinging. All statistical analyses were significant at p[lt]0.05.[br][bold]Results: [/bold]A total of 295 respondents (60 patients; 235 caregivers, 234 who were proxies for patients [lt]13 years) were analyzed. There were 74 Norditropin[sup][reg][/sup] FlexPro[sup][reg] [/sup]and Norditropin NordiFlex[sup][reg][/sup] users compared with 221 for other products. Overall mean age of patients on Norditropin[sup][reg] [/sup]vs. other was 13.2[plusmn]13.9 and 16.0[plusmn]14.7 years (p=0.01). The majority of rhGH patients/caregivers reported the use of a pen device (N=214) compared to vial (N=52). Significantly lower proportions of Norditropin[sup][reg] [/sup]users experienced any pain compared to others (43.3% vs. 65.8%, p=0.01) in bivariate analysis. Similar results were observed for stinging (55.4% vs.76.3%, p[lt]0.01). The significant association of Norditropin[sup][reg][/sup] use with reduced pain and stinging persisted in multivariate analyses adjusting for respondent type, patient age, device type, product and number of missed doses in the past month. Norditropin[sup][reg][/sup] users were 71.2% less likely to agree that injections are painful (95% CI 0.15-0.56) and 72.0% less likely to agree that injections cause stinging (95% CI 0.14-0.54) compared to others. Caregivers were three and a half times more likely to agree that injections are painful than patients (95% CI 1.46-5.74) though this significance did not persist in the stinging analysis. Those who reported a greater number of missed doses in the previous month were more likely to indicate injections were painful (p[lt]0.01). Patient age was not significant in either model.[br][bold]Conclusion: [/bold]Differences in the experience of pain and stinging were observed across rhGH products, though the influence of caregivers responding as proxies for patients warrants further investigation. Patients may prefer a product that is associated with less pain and stinging, thereby increasing medication adherence.[br][br]Sources of Research Support: This research was funded by Novo Nordisk Inc.[br][br]Disclosures: VD: Researcher, Novo Nordisk. T-MD: Employee, Novo Nordisk. JG: Employee, Novo Nordisk. SC: Researcher, Novo Nordisk. PA: Researcher, Novo Nordisk. LCW: Researcher, Novo Nordisk. WT: Employee, Novo Nordisk. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1587 198 1619 SUN-607 PO29-02 Sunday 1416 2012


1413 ENDO12L_SUN-608 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Evaluation of Glucose Homeostasis in a Large Cohort of Children with Growth Hormone (GH) Deficiency (GHD): Results from a Prospective, Case-Control Study Donatella Capalbo, Lucia De Martino, Andrea Esposito, Flavia Barbieri, Nicola Improda, Manuela Cerbone, Raffaella Di Mase, Mariacarolina Salerno University Federico II, Naples, Italy [bold]Background: [/bold]Adults with untreated GH deficiency (GHD) may have a cluster of cardiovascular risk factors. The effects of GH replacement therapy on insulin homeostasis in GHD subjects are still debated. Only a few studies investigated the effects of GHD and GH therapy on glucose metabolism in children.[br][bold]Objective[/bold][bold]: [/bold]To evaluate the effects of GHD and GH treatment on glucose metabolism in a large cohort of GHD children before and after GH replacement therapy.[br][bold]Subjects and methods: [/bold]Fasting glucose, insulin, HbA1c, and HOMA were assessed in 60 GHD children, aged 9.8[plusmn]0.3 years, before and after 1, 2 and 4 years of GH therapy. 60 healthy, age-, sex- and BMI-matched healthy controls were enrolled.[br][bold]Results[/bold][bold]: [/bold]In GHD children at baseline, fasting glucose (77.8[plusmn]1 vs 77.9[plusmn]1.2, mg/dl), insulin (4.7[plusmn] 0.4 vs 4.6[plusmn]0.4 [mu]U/ml), HOMA (1.26[plusmn]0.2 vs 1.12[plusmn]0.1) and HbA1c (5.29[plusmn]0.1 vs. 5.29[plusmn]0.04%) levels were comparable to healthy controls. One year of GH therapy (33[mu]g/kg/d) was associated with significant increase in insulin (7.69 [plusmn] 0.6, p [lt]0.0001) and HOMA (1.64 [plusmn] 0.15, p = 0.009), without significant changes in fasting glucose and HbA1c levels. Insulin and HOMA levels did not further increase after 2 (7.56[plusmn]0.6 and 1.85[plusmn]0.2, respectively) and 4 years (7.65[plusmn]0.7 and 1.5[plusmn]0.1, respectively) of treatment, remaining significantly elevated compared to pre-treatment levels. Fasting glucose and HbA1c did not change after 2 and 4 years of GH therapy.[br][bold]Conclusions: [/bold]Untreated GHD was not associated with significant alterations of glucose homeostasis. One year of GH treatment was associated with a slight impairment in insulin sensitivity without further change during long-term GH therapy.[br][br]Nothing to Disclose: DC, LDM, AE, FB, NI, MC, RDM, MS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1237 198 1620 SUN-608 PO29-02 Sunday 1417 2012


1414 ENDO12L_SUN-609 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Hemoglobin A1c during Growth Hormone Treatment in Prepubertal Children Molly O Regelmann, Lindsey Giserman, Elizabeth Chacko, Evan Graber, Ahmed Khattab, Dennis Chia, Michelle Klein, Elizabeth Wallach, Rachel Annunziato, Robert Rapaport Mount Sinai School of Medicine, New York, NY; Mount Sinai School of Medicine, New York, NY; Fordam University, Bronx, NY Background: Growth hormone (GH) treatment (GHT) has been associated with an increase in insulin resistance and rarely hyperglycemia and diabetes mellitus (DM) (1,2,3,4,5). Revised standards for assessing risk of developing DM emphasize hemoglobin A1c (HbA1c) as a screening tool (6). Trending HbA1c during GHT may be important for assessing risk of developing DM.[br]Hypothesis: In children with growth failure, GHT increases HbA1c.[br]Methods: Retrospective chart review of prepubertal children on GHT. Sex, ethnicity, diagnosis, family history, GH dose, insulin-like growth factor-1 (IGF-1), BMI Z score, height (Ht) and HbA1c were collected before or within 6 mo of starting GHT and at follow up every 3-6 mo, up to 2 yr in those without puberty. Paired t tests were performed to assess for rise in HbA1c on GHT. Pearson correlation tests were used to evaluate association of change in HbA1c with change in IGF-1, Ht standard deviation (SD) and BMI Z score.[br]Results: 38 patients (22 male, 16 female), age 7.4[plusmn]2.5 yr at start of GHT were reviewed. 21 had GH deficiency (stimulated GH peak[lt]10 ng/mL), 12 GH peak[ge]10 ng/mL, 3 small for gestational age, 2 Turner syndrome. 6 patients had a family history of type 2 DM. 34 were Caucasian, 3 Hispanic, 1 African-American. Initial mean Ht SD was -2.7[plusmn]0.7 and BMI Z score -0.1[plusmn]1.2. Mean dose of GH was 0.24[plusmn]0.04 mg/kg/wk. 19 patients had a pre-GHT HbA1c mean of 5.2[plusmn]0.3%. 19 patients had the initial HbA1c measured in the 1st yr on GHT with a mean of 5.1[plusmn]0.3%. Mean increase in HbA1c from initial to last HbA1c for all patients was 0.12% (p=0.09) and among males HbA1c increased 0.16% (p=0.016). An increase in HbA1c by 0.15% was noted at 13-18 mo (n=22, p=0.045) but no significant increases at 7-12 mo and 19-24 mo were noted when compared to initial HbA1C. The change in IGF-1, Ht SD and BMI Z score while on treatment did not significantly correlate with change in HbA1c.[br]Conclusions: The data presented show a trend toward a minor and clinically insignificant increase in HbA1c while on GHT. The HbA1c increase at 13-18 mo after initiation of GHT appears to have been transient. To further elucidate the relationship between GHT and HbA1c, additional data in those treated for longer duration and in pubertal children should be evaluated.[br][br](1) Husbands S, et al. Increased insulin sensitivity in young, growth hormone deficient children. Clin Endocrinol. 2001 Jul;55(1):87-92. (2) Riedl M, et al. The increased insulin sensitivity in growth hormone-deficient adults is reduced by growth hormone replacement therapy. Eur J Clin Invest. 2000 Sep;30(9):771-8. (3) Cutfield WS, et al. Safety of growth hormone treatment in children born small for gestational age: the US trial and KIGS analysis. Horm Res. 2006;65 Suppl 3:153-9. (4) Woodmansee WW, et al. Occurrence of impaired fasting glucose in GH-deficient adults receiving GH replacement compared with untreated subjects. Clin Endocrinol. 2010 Jan;72(1):59-69. (5) Sadeghi-Nejad A. Development of diabetes mellitus in two boys after the initiation of growth hormone therapy. J Pediatr Endocrinol Metab. 2007 Apr: 20(4): 541-4. (6) Executive Summary: Standards of Medical Care in Diabetes [ndash] 2010. Diabetes Care. 2010 Jan; 33,Suppl 1:S4-10.[br][br]Nothing to Disclose: MOR, LG, EC, EG, AK, DC, MK, EW, RA, RR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1756 198 1621 SUN-609 PO29-02 Sunday 1418 2012


1415 ENDO12L_SUN-610 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Characteristics of Pediatric Patients Treated with Growth Hormone and Prescribed Aromatase Inhibitor Therapy: Analysis of Data from the ANSWER Program[reg] Judith L Ross, Peter A Lee, Robert Z Gut, Vatsala Karwe, John A Germak Thomas Jefferson University, DuPont Hospital for Children, Wilmington, DE, Philadelphia, PA; Pennsylvania State College of Medicine, The Milton S Hershey Medical Center, Hershey, PA; Novo Nordisk, Inc, Princeton, NJ [bold]Background and Objective:[/bold] The American Norditropin Studies: Web-enabled Research (ANSWER) Program[reg], a US-based registry, has collected efficacy and safety information on patients treated with Norditropin[reg] (somatropin rDNA origin, Novo Nordisk A/S, Bagsvaerd, Denmark) enrolled at the discretion of participating physicians. In patients who were prescribed an aromatase inhibitor (AI) by their physicians, baseline characteristics and clinical data before and following growth hormone (GH) treatment, up to the time of AI initiation, were collected.[br][bold]Method and Results:[/bold] As of October 2011, 57 male GH-naive patients with GH deficiency (GHD, isolated, n=32; multiple pituitary hormone deficiency, n=2), idiopathic short stature (ISS, n=17), and patients with other growth disorders (n=6) including small for gestational age, Noonan syndrome, and Prader-Willi syndrome, were prescribed an AI (anastrozole or letrozole) during GH treatment. For the diagnostic categories, baseline mean ([plusmn]SD) chronologic age (CA) was 12.5[plusmn]3.2 y for GHD and 12.1[plusmn]3.3 y for ISS. At the start of AI treatment (AIT), mean CA was 15.1[plusmn]1.8 y for GHD and 15.5[plusmn]2.0 y for ISS. Of 57 patients, the number of patients classified as Tanner stage II, III, IV, or V was 6, 15, 8, and 25, respectively. Mean ([plusmn]SD) height standard deviation score (HSDS) at baseline and at AIT for all 3 groups was as follows: GHD: baseline, [minus]1.9[plusmn]1.0; AIT: [minus]1.0[plusmn]0.9; ISS: baseline, [minus]2.3[plusmn]0.6; AIT: [minus]1.1[plusmn]0.6; other: baseline, [minus]2.6[plusmn]0.3; AIT: [minus]1.6[plusmn]0.3. Mean ([plusmn]SD) bone age (BA) increased from baseline to AIT as follows: GHD: baseline, 11.7[plusmn]2.5 y; AIT: 13.8[plusmn]1.2 y; ISS: baseline, 10.4[plusmn]3.6 y; AIT: 13.6[plusmn]1.2 y. Mean ([plusmn]SD) BA/CA ratios did not change from baseline (0.88[plusmn]0.14, n=44) to AIT for all patients (0.91[plusmn]0.12, n=19) including GHD patients (0.91[plusmn]0.14, n=11) and ISS patients (0.92[plusmn]0.11, n=6). In comparison with the overall registry populations for GHD (n=5460) and ISS (n=1096), mean ([plusmn]SD) baseline CA for GHD and ISS males receiving AI were 1-2 years older (Overall: GHD: 10.6+3.9 y; ISS: 11.2+3.1 y). Mean ([plusmn]SD) baseline BA/CA ratios for the overall population were 0.83[plusmn]0.16 for GHD (n=4317) and 0.85[plusmn]0.14 for ISS (n=906).[br][bold]Conclusions: [/bold]AI treatment was initiated in older GHD and ISS patients nearly 2 years after GH treatment, coincident with advanced Tanner stage and bone age, suggesting their physicians were concerned about a diminishing treatment window for stimulating growth due to skeletal maturation and epiphyseal fusion.[br][br]Disclosures: JLR: Advisory Group Member, Novo Nordisk, Eli Lilly & Company, Abbott Laboratories; Clinical Researcher, Novo Nordisk, Pfizer, Inc., Eli Lilly & Company. PAL: Advisory Group Member, Abbott Laboratories, Novo Nordisk; Clinical Researcher, Abbott Laboratories, Eli Lilly & Company, Pfizer, Inc., Ipsen. RZG: Employee, Novo Nordisk. VK: Employee, Novo Nordisk. JAG: Employee, Novo Nordisk. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2307 198 1622 SUN-610 PO29-02 Sunday 1419 2012


1416 ENDO12L_SUN-611 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Mortality in Growth Hormone (GH)-Treated Pediatric Patients Enrolled in a Global Prospective Observational Study Christopher J Child, Alan G Zimmermann, Charmian A Quigley, Ron G Rosenfeld, Leslie L Robison, Werner F Blum Eli Lilly and Company, Windlesham, UK; Oregon Health [amp] Science University, Portland, OR; St Jude Children[apos]s Research Hospital, Memphis, TN Preliminary data from the French SAGhE study follow-up of 6,928 adults who started GH treatment in childhood for isolated idiopathic GH deficiency (IsIGHD), idiopathic short stature (ISS) or short stature following small for gestational age (SGA) birth, suggested increased mortality versus the French general population (standardized mortality ratio [SMR]: 1.3, 95% confidence interval [CI]: 1.1-1.6, 116,403 person-years [PY])[sup](1)[/sup].[br]We assessed mortality in the global Genetics and Neuroendocrinology of Short Stature International Study (GeNeSIS) pediatric observational program. Deaths in GeNeSIS patients (pts; all diagnoses, including organic and syndromic causes of short stature) were identified; for reference to SAGhE data, mortality rates were calculated for GH-treated (tx) pts with IsIGHD, ISS and SGA. SMR and 95% CI were calculated using age- and sex-specific population rates from CDC (US, http://wonder.cdc.gov) and WHO (other countries, http://apps.who.int/ghodata).[br]33 deaths were reported among 18,147 pts (30/17,692 GH-tx, 3/455 non-GH-tx) during median (Q1, Q3) follow-up of 2.0 (0.9, 3.8) years (y). Deaths were due to acute illness (n=16), recurrence of pre-existing intracranial neoplasms (n=8, including the 3 non-GH-tx pts), second cancers (n=2, both irradiated), accidents (n=3), underlying conditions (n=2) and unknown causes (n=2).[br]5 deaths were reported among 11,076 GH-tx pts with diagnoses of IsIGHD, ISS and SGA [25,634 combined PY of follow-up in GeNeSIS, crude mortality rate (95% CI): 19.5 (6.3-45.5)/100,000 PY, SMR (95% CI): 0.3 (0.1-0.8)]. The combined mortality rate in pts with IsIGHD, ISS and SGA in GeNeSIS was similar to the 2007 US age-adjusted mortality rate for children aged 5-14 years (15.3/100,000 PY)[sup](2)[/sup]. For 7,712 pts with IsIGHD there were 3 deaths, SMR (95% CI): 0.3 (0.1-0.7); for 2,538 pts with ISS 1 death, 0.5 (0.0-2.6); and for 826 pts with SGA 1 death, 0.7 (0.0-4.0). SMRs for the diagnoses of interest, both individually and combined, were not elevated.[br]There was no evidence of increased mortality in the GH-tx IsIGHD, ISS or SGA pts in GeNeSIS. However, our analysis in pts during GH treatment is not directly comparable with SAGhE, which assessed mortality rates many years after GH treatment. Additional limitations include comparison with general population data (due to lack of non-GH-tx controls) and limited follow-up time.[br][br](1) Carel J-C et al., J Clin Endocrin Metab. First published ahead of print January 11, 2012 as doi:10.1210/jc.2011-1995. (2) Xu J et al., National Vital Statistics Reports 2010; 58(19).[br][br]Sources of Research Support: Funded by Eli Lilly and Company.[br][br]Disclosures: CJC: Employee, Eli Lilly & Company. AGZ: Employee, Eli Lilly & Company. CAQ: Employee, Eli Lilly & Company. RGR: Scientific Board Member, Eli Lilly & Company; Speaker, Eli Lilly & Company. LLR: Scientific Board Member, Eli Lilly & Company; Speaker, Eli Lilly & Company. WFB: Employee, Eli Lilly & Company. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 391 198 1623 SUN-611 PO29-02 Sunday 1420 2012


1417 ENDO12L_SUN-612 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) The Impact of Exercise-Stimulated Growth Hormone on Exercise-Induced Markers of Anti-Oxidation and Pro-Oxidation in Pre- and Early Pubertal Normal Weight and Overweight Boys George Paltoglou, George Valsamakis, Maria Shina, Antonis Kampas, Athanasios Chantzinikolaou, Aimilia Mantzou, Ioannis Papassotiriou, George P Chrousos, Christina Kanaka-Gantenbein, Ioannis G Fatouros, George Mastorakos Medical School, University of Athens, Athens, Greece; Medical School, University of Athens, Athens, Greece; University of Thrace, Komotini, Greece; Medical School, University of Athens, Athens, Greece; [quot]Aghia Sofia[quot] Children[apos]s Hospital, Athens, Greece Exercise is a potent stress stimulus, with multiple adaptive hormonal, cytokine and biochemical responses. Moreover, obesity predisposes to oxidative/anti-oxidative status imbalance.[br]To examine the impact of exercise-induced hormonal stimulation upon the anti-oxidation and pro-oxidation markers in normal and overweight, pre- and early pubertal boys, 60 healthy untrained male children aged 10.81 [plusmn] 0.99 (mean[plusmn]SD) years were recruited.The protocol consisted of two sessions scheduled two weeks apart. In the first session, participants had their maximal aerobic capacity (VO2max) assessed. In the second, they underwent a baseline blood collection and then performed an exercise bout corresponding to the 70% of VO2max. Immediately thereafter a second blood sampling was undertaken.[br]Measurements included the pro-oxidation products: Thiobarbituric Acid Reactive Substances (TBARS) and Protein Carbonyls (PCs), the anti-oxidation products: glutathione (GSH), oxidized glutathione (GSSG), the antioxidant enzymes Catalase and Glutathione Peroxidase (GPX) and the total antioxidant capacity (TAC), the hormones: FSH, LH, Testosterone, ACTH, Cortisol and Growth Hormone (GH) the inflammatory markers: interleukin-6 (IL-6) and C reactive protein (CRP) and the exercise variable: VO2max.[br]Baseline oxidative stress markers were significantly greater in overweight pre-pubertal subjects, compared with the respective normal weight group. Post-exercise oxidative stress markers increased significantly in all groups. Post-exercise GH levels significantly increased (P[lt]0.05) in normal and overweight pubertal subjects, but not in pre-pubertal ones. The change ([Delta]) of GH (G[Eta]) showed a statistically significant positive linear correlation with [Delta]PC and [Delta]GSSG only in pubertal normal weight subjects. When a forward stepwise regression model was applied to define potential predictors of post exercise anti-oxidation and pro-oxidation markers, baseline GH was found to be a potential predictor of the post-exercise anti-oxidation markers GSH (positive), GSH/GSSG (positive), GPX (negative) and the pro-oxidation marker PCs (negative, along with VO2max). Furthermore a forward stepwise regression model revealed baseline GSSG as the best predictor of VO2max.[br]GH increase seems to play an important role in oxidative stress markers modification following 70% VO2max aerobic exercise in puberty. The anti-oxidation stress marker GSSG is the best predictor of VO2max, while the other variables tested were not.[br][br]Nothing to Disclose: GP, GV, MS, AK, AC, AM, IP, GPC, CK-G, IGF, GM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1832 198 1624 SUN-612 PO29-02 Sunday 1421 2012


1418 ENDO12L_SUN-613 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Arginine and Levo-Dopa Stimulation in Children: Association of Peak Growth Hormone Response with Homeostatic Model Assessment and Lipids Elizabeth Chacko, Evan Graber, Elizabeth Wallach, Molly Regelmann, Rachel Annunziato, Michelle Klein, Dennis Chia, Ahmed Khattab, Gertrude Costin, Robert Rapaport Mount Sinai School of Medicine, New York, NY [bold]Background: [/bold]Growth hormone (GH) stimulation testing (ST) is part of the evaluation of growth failure in children. Insulin resistance (IR) and lipid abnormalities have been reported in GH deficient (GHD) adults. In GHD children, positive and negative correlations between peak GH response (PGH) and IR have been found. We are not aware of studies relating PGH with IR and lipids in a population of unselected children.[br][bold]Objective: [/bold]To correlate PGH levels in response to arginine and levo-dopa (ALD) ST with Homeostatic Model Assessment (HOMA-IR) and lipids.[br][bold]Design/Methods: [/bold]Retrospective chart review of children with growth failure who underwent ALD ST. Data collection included age, sex, height, weight, pubertal status, HbAIc, fasting preserved glucose, insulin, c-peptide and lipid profile. GH levels were measured by Esoterix Lab (Calabasas Hills, CA). HOMA-IR was calculated by standard formula [HOMA-IR (mmol/L x [micro]U/ml) = fasting glucose (mmol/L) x fasting insulin ([micro]U/ml)/22.5]. Statistical analyses included Pearson correlations and t-tests.[br][bold]Results: [/bold]Data of 132 (89 M) consecutively tested children (mean age 10.5[plusmn]2.9 yr; BMI Z-score -0.176 [plusmn] 1.02) were reviewed. Group1 (n=62) had PGH[lt]10 ng/mL and Group 2 (n=70) had PGH =/[gt] 10 ng/mL. IGF-1 was lower in group I (p=0.022). HOMA-IR and HbAIc were normal in all patients with no difference between groups 1 and 2. HOMA-IR was higher in group 1 verses group 2 in prepubertal females (PPF) (p=0.022) and pubertal males (PM) (p=0.007). No overall differences were seen in lipids between groups 1 and 2. In group1 versus group 2, total cholesterol (TC) was higher in PM (p=0.034) and lower in all prepubertal (PP) (p=0.008) children. LDL-C was lower in group 1 PPF (p=0.040) subjects. In all patients, PGH negatively correlated with HOMA-IR (r=-0.224; p=0.032) and in subgroups PP (r= -0.380; p=0.006) and PPF(r=-0.617; p=0.008) subjects. PGH negatively correlated with TC in all patients (r=-0.289; p=0.015) and especially so in pubertal females (PF) (r=-0.834; p=0.001). PGH negatively correlated with triglycerides only in PP M (r=-0.458; p=0.024) and with LDL-C only in PF (r= -0.755; p=0.007) subjects.[br][bold]Conclusion:[/bold] In this group of non-obese, non insulin resistant children, HOMA-IR and lipids negatively correlated with PGH response to ALD ST. These findings suggest an effect of glucose, insulin, and lipids on growth hormone secretory dynamics.[br][br]Nothing to Disclose: EC, EG, EW, MR, RA, MK, DC, AK, GC, RR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1758 198 1625 SUN-613 PO29-02 Sunday 1422 2012


1419 ENDO12L_SUN-614 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Prevalence of Primary and Secondary IGF-I Deficiency in Short Children Tested for Growth Hormone Deficiency Walter Bonfig, Susanne Bechtold, Stephanie Putzker, Claudia Weissenbacher, Heinrich Schmidt, Hans P Schwarz Children[apos]s Hospital of the Technical University, Munich, Germany; Children[apos]s Hospital of Ludwig-Maximilians University, Munich, Germany Background: The prevalence of primary IGF-I deficiency is not well known. However, identifying children with this condition is important as some may profit from rhIGF-1 treatment.[br]Objective: To assess the prevalence of primary IGF-I deficiency (defined as height SDS[lt]-3.0 and circulating IGF-I SDS[lt]-2.0) after exclusion of classic growth hormone deficiency (GHD) in a large group of short children and adolescents seen at one pediatric endocrine referral center.[br]Patients: Between 2004 and 2009, growth hormone stimulation tests with clonidine (n=353) and arginine (n=131) were performed in a cohort of 358 patients (258 males, 100 females) being referred and evaluated for short stature and suspected GHD.[br]Results: Classic GHD based on 2 pharmacological stimulation tests with a maximum GH level [lt]8 ng/dl in both was diagnosed in 124/358 patients (34.6%; 82 males, 34 females). Among the remaining 234 patients there were 109 patients with an IGF-I SDS[lt]-2.0. Full clinical and anthropometric data were available in 67 patients. Of those, 38 (20 males, 18 females) had a height SDS[lt]-3.0 (mean+/-SD: -3.9+/-1.2) and an IGF-I SDS[lt]-2.0 (-2.5+/-0.3). Mean age of this group was 9.1+/-3.9 years (range: 3.2-15.2) thus fulfilling the criteria of primary IGF-I deficiency. The clinical diagnosis in these children had been idiopathic short stature as systemic, skeletal and syndromic disease had mostly been excluded.[br]Conclusion: In the setting of a large outpatient pediatric clinic the prevalence of primary IGF-I deficiency was about one third of the prevalence for GHD. With stringent criteria used for the indication of GH stimulation testing about half of the short children with IGF-I SDS [lt]-2.0 may have primary IGF-I deficiency. Careful documentation and follow-up of this group of patients is certainly warranted as a treatment option with rhIGF-1 has become available.[br][br]Nothing to Disclose: WB, SB, SP, CW, HS, HPS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1975 198 1626 SUN-614 PO29-02 Sunday 1423 2012


1420 ENDO12L_SUN-615 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Outcomes in Children with Growth Disorders Treated with Recombinant Human Insulin-Like Growth Factor-1 (IGF-1) Barry J Reiner, Brad S Miller, Joyce Kuntze, Don Carver, Sandra L Blethen University of Minnesota Amplatz Children[apos]s Hospital, Minneapolis, MN; Ipsen, Brisbane, CA; , Wherever, Maryland California [bold]Background: [/bold]The Increlex[reg] (mecasermin [rDNA origin] injection) Growth Factor Database (IGFD) began in 05/2006 to collect long term data on safety and efficacy in children treated with Increlex. As of 05/2011, there were 1093 patients with completed enrollment forms and at least one follow-up visit. We explored factors that might affect safety and growth outcomes. Because total height gain ([Delta]Ht SDS) in children receiving growth hormone is dependent on [Delta]Ht SDS before the onset of puberty(1), we studied [Delta]Ht SDS before puberty as well as first-year height-velocity (HV).[br][bold]Methods:[/bold] All patients are included in the safety analysis. First year HV was studied in the 154 children who were prepubertal throughout their first year of treatment. [Delta]Ht SDS was studied in children who were prepubertal when they began treatment and attained Pubertal stage 2 while receiving Increlex[reg] (n= 181). The following baseline variables were explored in multiple regression analyses: age, IGF-1 SDS, Ht SDS, BMI SDS, average cumulative dose, mother[apos]s height SDS, father[apos]s height SDS, and gender. Duration of treatment before the onset of puberty was tested in the analysis of total prepubertal growth. P-values [lt] 0.01 were considered to be significant.[br][bold]Results: [/bold]There were 197 patients (18.0%) who reported any treatment emergent adverse event (AE). Aes were typically those listed in the Increlex prescribing information. Hypoglycemia (n=100, 9.1%), headaches (n=64, 5.9%), and injection site reactions (n=19, 1.7%) were the most common Aes. Intracranial hypertension occurred in 9 children (0.8%) Within the range of Increlex[reg] doses reported (37-170 [micro]gms/kg/bid), no AE was dose-dependent. Mean HV in patients who remained prepubertal was 7.1 [plusmn] 1.7 cm/y). HV was negatively related to age at baseline (p[lt]0.0001) and positively related to cumulative average Increlex[reg] dose (p=0.0002). [Delta]Ht SDS before puberty was negatively related to age and Ht SDS at baseline (p[le]0.0001) and positively related to duration of treatment before pubertal stage 2 (p[lt]0.0001).[br][bold]Conclusions: [/bold]The pattern and frequency of Aes in the registry remain unchanged. There have been no new safety signals. Beginning treatment at a younger age not only improves first-year HV, but allows for a longer period of prepubertal treatment that should eventually result in greater adult height gain. For the first year of growth, beginning treatment at younger ages with appropriate weight-adjusted doses was important for better outcomes.[br][br](1) Blethen SL, Baptista J, Kuntze J, Foley T, LaFranchi S, Johanson A. Adult height in growth hormone (GH)-deficient children treated with biosynthetic GH. J Clin Endocrinol Metab 1997;82:418-420.[br][br]Sources of Research Support: Ipsen.[br][br]Disclosures: BJR: Study Investigator, Ipsen; Advisory Group Member, Ipsen. BSM: Consultant, Genentech, Inc., Pfizer, Inc, Novo Nordisk, Eli Lilly & Company, Tercica, Sandoz, Teva, Endo Pharmaceuticals Inc.; Investigator, Genentech, Inc., Pfizer, Inc., Novo Nordisk, Eli Lilly & Company; Tercica, Endo Pharmaceuticals Inc., Abbott Laboratories; Advisory Group Member, Ipsen; JK: Employee, Ipsen. DC: Consultant, Ipsen. SLB: Consultant, Ipsen. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 837 198 1627 SUN-615 PO29-02 Sunday 1424 2012


1421 ENDO12L_SUN-616 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) The Relationship between Phalangeal Length and Height or Growth Velociy Young Seok Shim, Il Tae Hwang, Seung Yang Kangdong Sacred Heart Hospital, Seoul, Korea Purpose: There is a positive correlation in growth among long bones of body. Height will reflect the length of femur and tibia. We want to know how to predict growth or final height with phalangeal length.[br]Subjects and methods: Total 52 children, 35 boys and 17 girls, who visited our clinic in last 2 years and checked bone age and revisited at least once more after the 1st visit were included in this study. The patient who had the disease related with growth such as growth hormone deficiency and precocious puberty were excluded. We reviewed their charts retrospectively. We measured the phalangeal length (the proximal phalanx of the 3rd finger) from the X-ray film (left hand AP) at the 1st visit.[br]Results: The age is 9.1[plusmn]2.7 year (boys, 9.0[plusmn]3.0 year, girls, 9.1[plusmn]1.8 year). Interval between the 1st visit and the next visit is 1.0[plusmn]1.0 year (boys, 1.2[plusmn]1.2 year, girls, 0.7[plusmn]0.4 year). The growth velocity between the 1st visit and the next visit is 6.6[plusmn]2.5 cm/year (boys, 6.5[plusmn]2.7 cm/year, girls, 6.9[plusmn]2.0 cm/year). There is a positive correlation between height and proximal phalangeal length of the 3rd finger (P=0.000, R=0.920). There is no correlation between growth velocity and proximal phalangeal length of the 3rd finger (P=0.073, R=0.608).[br]Conclusions: As there is a positive correlation between phalangeal length and height, all long bones may have propotional growth. We couldn[apos]t show the correlation between phalangeal length and growth velocity, so we should consider many other factors such as bone age and parental height in prediction of the growth and final height.[br][br]Nothing to Disclose: YSS, ITH, SY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1538 198 1628 SUN-616 PO29-02 Sunday 1425 2012


1422 ENDO12L_SUN-617 POSTER SESSION: Growth Hormone [amp] Growth Disorders (1:30 PM-3:30 PM) Mild GH Deficiency Due to Two Novel Homozygous Mutations in the Gene Encoding Growth-Hormone Releasing Hormone Receptor ([italic]GHRHR[/italic]) in a Single Family Louise Cheryl Gregory, Kyriaki S Alatzoglou, Mark J McCabe, Paul Letissier, Mehul T Dattani UCL Institute of Child Health, London, UK The release of Growth Hormone (GH) by the somatotroph cells of the anterior pituitary is stimulated by GH releasing hormone (GHRH) released from the hypothalamus. GHRH acts via its transmembrane receptor, GHRHR, a G-protein coupled receptor that stimulates protein kinase A. Recessive [italic]GHRHR[/italic] mutations are associated with severe isolated GH deficiency (IGHD Type 1B), with an untreated final height of 130cm[plusmn]10cm (-7.2[plusmn]1.6SDS) and 114[plusmn]0.7cm (-8.3[plusmn]0.1SDS) in males and females respectively. The characteristic phenotype is not classical of IGHD in that patients often present minimal facial hypoplasia, no hypoglycaemia or microphallus and variable anterior pituitary hypoplasia on magnetic resonance imaging (MRI). We hypothesised that a consanguineous Pakistani family with IGHD in 3 siblings (2 males, 1 female) would have mutations in [italic]GH1[/italic] or [italic]GHRHR[/italic]. PCR amplification and direct sequencing analysis were used to screen both genes. In all three affected siblings we identified two novel homozygous missense mutations [c.11G[gt]A (p.R4Q), c.236C[gt]T (p.P79L)], absent from 200 Pakistani controls, in a conserved region of the extracellular domain of [italic]GHRHR[/italic]. Both are predicted to affect protein conformation and ligand binding with a protein prediction model predicting p.P79L to be functionally deleterious[italic]. [/italic]The brothers were diagnosed with GHD at the age of 9.8 and 6.0 years with a height SDS of -2.24 and -1.23 respectively (mid parental height of -1.10 SDS). The peak GH concentrations to glucagon stimulation were 2.9[mu]g/l with low IGF-1 and IGFBP3. Both brothers presented with micropenis, hypoplastic scrotum and bilateral undescended testes. Their elder sister presented at 16 years with untreated isolated GHD (peak GH to insulin tolerance test [lt]0.1[micro]g/l, IGF-1[lt]3.3mmol/L). She had classic GHD with abdominal fat deposition, a high-pitched voice and frontal bossing. Surprisingly, she attained an untreated final height of 144 cm (-3.0 SDS), which is the tallest untreated height reported in a patient with a [italic]GHRHR[/italic] mutation. All 3 siblings had a small anterior pituitary on MRI. Their asymptomatic mother was compound heterozygous for both mutations; paternal DNA is currently unavailable. Screening of [italic]GH1[/italic] revealed no mutations. Functional analysis of the two mutations is currently underway. We report the presence of two novel homozygous mutations in [italic]GHRHR[/italic] in a single pedigree. The phenotype in this family appears to be relatively mild despite the presence of two mutations in the same gene.[br][br]Nothing to Disclose: LCG, KSA, MJM, PL, MTD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 380 198 1629 SUN-617 PO29-02 Sunday 1426 2012


1423 ENDO12L_SUN-618 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Phenotype[ndash]Genotype Correlations in Patients with Inactivating Gonadotropin Hormone Receptor Mutations Daiane Beneduzzi, Ericka B Trarbach, Elaine MF Costa, Ana Paula Abreu, Adriana Lofrano-Porto, Berenice Bilharinho Mendonca, Ana Claudia Latronico, Leticia G Silveira HC/FMUSP, S[atilde]o Paulo, Brazil; HUB/FS/UnB, Bras[iacute]lia, Brazil Context: Loss-of-function GnRH receptor mutations (GNRHR) cause normosmic congenital isolated hypogonadotropic hypogonadism (nIHH). Aim: To characterize the frequency and phenotype-genotype correlation of GNRHR mutations. Patients and methods: GNRHR mutational analysis, detailed phenotypic characterization and long term follow-up were performed in 86 Brazilian patients (58 males and 28 females), including 16 familial cases (18.6%). Results: Six GNRHR mutations were identified (p.N10K; p.Q11K; p.Q106R p.R139H; p.C200Y, p.Y283H) in 7 patients (8,1%), 4 males and 3 females, and were more prevalent in familial cases (25%) than in sporadic cases (4,3%). Mutations in other genes (GNRH1, PROK2, PROKR2, TAC3, TAC3R, KISS1 and KISS1R) were present in 17.4% of all patients and no digenic defects were identified. No phenotypic difference was observed between patients with and without GNRHR mutations. All males had micropenis (mean penile length: 5.6 cm) and cryptorchidism was diagnosed in 23% of them. Among patients with GNRHR mutations 4 had homozygous mutations (p.R139H and p.C200Y) whereas 3 patients exhibited compound heterozygous mutations (p.[N10K; Q11K]; [Q106R], p.[N10K;Q11K]; [Y283H], p.[R139H];[Q106R]). The p.R139H mutation was the most prevalent defect on the Brazilian group. The p.R139H and p.C200Y mutations cause complete loss of function of the receptor in vitro and were associated with complete nIHH in homozygous patients, whereas the [p.N10K;p.Q11K] and the Q106R are partially inactivating mutations. The woman that was compound heterozygous for [p.N10K; p.Q11K] and p.Q106R had partial nIHH, with spontaneous thelarce and detectable levels of LH. The male who was compound heterozygous for the p.R139H and p.Q106R mutations had a sustained reversal of hypogonadism after discontinuation of hormonal therapy. Conclusion: GnRHR was the most commonly affected gene in nIHH and should be the first candidate for genetic analysis, particularly in autosomal recessive familial cases. A genotype-phenotype correlation was observed in nIHH patients with GNRHR mutations, considering that patients carrying partially inactivating mutations had either partial IHH or reversal of the hypogonadism.[br][br]Sources of Research Support: Financial support: FAPESP and CAPES.[br][br]Nothing to Disclose: DB, EBT, EMFC, APA, AL-P, BBM, ACL, LGS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1081 199 1630 SUN-618 PO29-01 Sunday 1427 2012


1424 ENDO12L_SUN-619 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Mutational Analysis of Neurokinin B and Its Receptor in Patients with Idiopathic Central Pubertal Disorders Cintia Tusset, Sekoni Noel, Leticia Silveira, Ericka Trarbach, Elena Gianetti, Stephanie Seminara, Berenice Bilharinho Mendonca, Ursula Kaiser, Ana Claudia Latronico Hospital das Clinicas da Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Brigham and Women[apos]s Hospital, Boston, MA; Massachusetts General Hospital, Boston, MA Neurokinin B (NKB) signaling appears to play a key role in human pubertal development. It is highly expressed in hypothalamic neurons that also express kisspeptin, a potent stimulator of GnRH secretion.[br]Aim: To investigate the presence of variants in the TAC3 and TACR3 genes, which encode NKB and its receptor (NK3R), respectively, in a large cohort of patients with idiopathic central pubertal disorders.[br]Patients and Methods: Two hundred and thirty seven patients were studied: 114 with central precocious puberty (CPP), 50 with constitutional delay of growth and puberty (CDGP), and 73 with normosmic isolated hypogonadotropic hypogonadism (IHH). The control group consisted of 150 Brazilian individuals with normal pubertal development. Genomic DNA was extracted from peripheral blood and the entire coding region of both TAC3 and TACR3 genes were amplified and automatically sequenced. In silico and in vitro analyses were performed. Other genes involved with central pubertal disorders, FGFR1, GNRHR, GNRH, KISS1, KISS1R, PROK2 and PROKR2, were previously studied.[br]Results: We identified one variant (p.A63P) in the NKB and four variants (p.G18D, p.L58L, p.W275X and p.A449S) in the NK3R, which were absent in the control group. All of them were found in the heterozygous state, except p.W275X which was identified in the homozygous state in an IHH patient. The new p.A63P variant was identified in a Brazilian girl with sporadic CPP. Her mother, who had normal pubertal development, was heterozygous for the same variant. The known p.G18D, p.L58L and p.W275X variants were identified in three unrelated males with complete IHH. All of them affected highly conserved residues. In silico analysis (PolyPhen and SIFT) predicted an impact of p.G18D substitution on protein structure, but in vitro studies showed that this variant did not impair NK3R signaling in transfected cells. The new p.A449S variant was identified in a Brazilian girl with CDGP, and in silico analysis suggested that this variant is benign.[br]Conclusion: Rare variants were identified in the TAC3 and TACR3 genes in patients with central pubertal disorders. Inactivating TACR3 mutations were associated with the normosmic IHH phenotype in our cohort.[br][br]Sources of Research Support: This work has been supported by FAPESP # 2005/04726-0 and CAPES.[br][br]Nothing to Disclose: CT, SN, LS, ET, EG, SS, BBM, UK, ACL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 807 199 1631 SUN-619 PO29-01 Sunday 1428 2012


1425 ENDO12L_SUN-620 POSTER SESSION: Puberty (1:30 PM-3:30 PM) A PROKR2 Mutant Exerts Dominant Negative Effects on Wild-Type PROKR2: New Insights into the Effects of PROKR2 Heterozygous Mutations Ana Paula Abreu, Shuyun Xu, Sekoni D Noel, Rona S Carroll, Ana Claudia Latronico, Ursula B Kaiser Brigham and Women[apos]s Hospital and Harvard Medical School, Boston, MA; HC/FMUSP, S[atilde]o Paulo, Brazil Background: Prokineticin 2 (PROK2) and prokineticin receptor 2 (PROKR2) play important roles in olfactory bulb neurogenesis and GnRH neuronal migration. Several mutations in PROKR2, located in diverse functional domains of the receptor, have been described in patients with varying degrees of GnRH deficiency, the great majority of which were identified in the heterozygous state. We previously identified the R80C PROKR2 mutation in the first intracellular loop (ICL1) of the receptor in the heterozygous state in a patient with Kallmann syndrome and showed that this mutation interferes with PROKR2 protein stability and proper localization in the plasma membrane. In this analysis, we sought to evaluate whether R80C PROKR2 exerts dominant negative effects on wild-type (WT) PROKR2 to contribute to the pathogenesis of GnRH deficiency.[br]Methods: We performed in vitro studies in HEK293 or COS7 cells by co-transfecting both WT and R80C PROKR2 in varying ratios and comparing responses to cells transfected with WT PROKR2 alone. To assess PROKR2 signaling activity we measured inositol phosphate (IP) accumulation. Binding studies evaluated PROKR2 specific ligand binding and Western blot analysis quantified PROKR2 plasma membrane density.[br]Results: IP studies showed a [sim]40% reduction in maximal PROK2-stimulated IP accumulation when equal amounts of WT and R80C PROKR2 were co-transfected, compared to WT PROKR2 alone, a response that was further reduced when the co-transfection ratio of R80C:WT PROKR2 was 4:1. Binding studies similarly showed a reduction in specific binding activity when WT and R80C PROKR2 were co-transfected compared to WT PROKR2 alone. Western blot analysis showed that co-transfection of R80C PROKR2 interfered with membrane expression of WT PROKR2. Taken together, these results indicate that the R80C PROKR2 mutant interferes with ligand binding and PROK2-activated signal transduction by the WT receptor through inhibitory effects on WT PROKR2 plasma membrane expression in vitro. These dominant negative effects were most evident when the mutant receptor was transfected in excess of the WT PROKR2.[br]Conclusions: We have shown that Arg80 in the ICL1 of PROKR2 is important for receptor stability and proper localization in the plasma membrane and that the R80C PROKR2 mutant exerts dominant negative effects on the WT receptor. The presence of R80C PROKR2 in the heterozygous state may thus contribute to hypogonadotropic hypogonadism through a dominant negative mechanism.[br][br]Nothing to Disclose: APA, SX, SDN, RSC, ACL, UBK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1606 199 1632 SUN-620 PO29-01 Sunday 1429 2012


1426 ENDO12L_SUN-621 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Mirror Movements in Female Carriers of a New Heterozygous Mutation in the X-Linked Kallmann Syndrome Gene (KAL1) Peter Gunczler, Laura G Gonzalez Briceno, Michel Polak, Livia Verdu, Nicolas de Roux Hospital de Clinicas Caracas, Caracas, Venezuela; H[ocirc]pital Necker [ndash] Enfants Malades, APHP Universit[eacute] Paris Descartes, Centre de Maladies Endocriniennes Rares de la Croissance, Paris, France; H[ocirc]pital Robert Debr[eacute], Paris, France Background: Kallmann Syndrome (KS) is a genetically heterogeneous developmental disease characterized by the association of hypogonadotropic hypogonadism, and hyposmia or anosmia. Additional findings may include mirror movements (bimanual synkinesis) but only described in males, as mutated female carriers are continuously considered as unaffected.[br]Objective: To correlate mirror movements, a frequently forgotten clinical feature of Kallmann Syndrome, and a gene defect in KAL1 in a family with hypogonadotropic hypogonadism and also to correlate the gene defect with a phenotype restricted to mirror movements.[br]Results: We describe herein the clinical heterogeneity in a family that comes from the North Eastern region of Venezuela who presented with a new complex change in KAL1 (c.[2067_2070AGGA[gt]TCCT; 2071delG]; p.Glu642Alafs21). Two affected brothers were found to be hemizygote for this substitution/deletion. The eldest child, a 13 y.o. boy, presented with unilateral cryptorchidism, small penis (4.5cm), gynecomastia, eunuchoid body habitus with an arm span exceeding height by 10 cm, he also had hyposmia and mirror movements noted since early childhood. The youngest child, a 6 y.o. boy, presented with micropenis, bilateral cryptorchidism, difficulty recognizing odors and mirror movements. The genetic analysis performed in their mother, who also has mirror movements, showed that she was a heterozygote carrier for the KAL1 genetic defect. The maternal grandmother was reported as also having mirror movements. In addition, several members of the maternal family, in both genders, had a history of mirror movements, and at least eight male relatives reported an association of micropenis and mirror movements. Another feature present in this family was the development of type 2 diabetes in association with KS in two of the maternal uncles of our patients.[br]Conclusion: Our findings show the presence of mirror movements in a large family presenting with Kallmann Syndrome due to a new KAL1 genetic defect. They were present not only in the affected males but also in some female heterozygote carriers. Anosmia and hypogonadotropic hypogonadism result from the abnormal migration of olfactory and GnRH neurons during brain development. This new genetic defect of KAL1 emphasizes the hypothesis that mirror movements in females may also be related to KAL1 loss of function mutation.[br][br]Nothing to Disclose: PG, LGGB, MP, LV, NdR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 622 199 1633 SUN-621 PO29-01 Sunday 1430 2012


1427 ENDO12L_SUN-622 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Long-Term Evaluation and Clinical Spectrum of the Pubertal Abnormalities of Two Siblings Harboring NR0B1 Inactivating Mutation Sorahia Domenice, Vinicius Nahime Brito, Priscila Cukier, Ivo Jorge Prado Arnhold, Mirian Yumi Nishi, Berenice Bilharinho Mendonca, Ana Claudia Latronico Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Congenital hypogonadotropic hypogonadism (HH) is associated with inactivating [italic]NR0B1 [/italic]mutations in patients with X-linked adrenal hypoplasia congenital (X-AHC). Normal or premature sexual development has also been reported in X-AHC patients, although the exact mechanisms involved in development of sexual precocity had not yet been elucidated. We previously described a boy with X-AHC due to a [italic]NR0B1[/italic] mutation (p.Cys65Leufs*6), who firstly presented with a gonadotropin-independent precocious puberty. Aim: To describe the long-term clinical and hormonal follow-up of this patient with X-AHC and his younger brother, also affected by the same mutation. Patients: The index case has been followed for 15 years (from 2.2 to 17 yrs). At 2.2 yrs of age the patient was evaluated due to clinical manifestation of isosexual precocious puberty. His hormonal analysis confirmed a gonadotropin-independent precocious puberty. Adrenal insufficiency was diagnosed and the steroid replacement therapy was introduced at 3 yrs of age. This therapy resulted in decrease of his testicular size and testosterone (T) levels to prepubertal range. Ciproterone acetate was added to steroid replacement therapy to achieve a better control of T levels at 11.5 yr. A progressive decline in T levels was observed after interruption of cyproterone acetate (at 14 yrs: T= 148 ng/dL; 17.4 yrs: T= 98 ng/dL), although his basal and GnRH-stimulated gonadotropin levels were at pubertal range. The lack of spontaneous pubertal progression was remarkable, indicating the diagnosis of HH. His affected younger brother, exhibited subclinical primary adrenal insufficiency in neonatal period and steroid replacement therapy was started. High gonadotropin and T levels confirmed a normal [ldquo]mini-puberty[rdquo] in this boy. At 11-mo, he presented a mild phenotype of premature sexual development (Tanner III), elevated ACTH (591 pg/mL) and T levels (143 ng/dL) and suppressed gonadotropin levels, confirming the diagnosis of gonadotropin-independent precocious puberty. Cyproterone acetate was added to steroid therapy at 1.75 yrs. Conclusion: The clinical and hormonal follow-up of the younger brother reinforce the hypothesis that excessive ACTH levels acted probably as an atypical stimulus to Leydig cells, causing gonadotropin-independent precocious puberty in this family. Additionally, the older brother showed a pubertal evolution to HH as typically observed in X-AHC patients despite the premature sexual development in his infancy.[br][br](1)Domenice S et al, J Clin Endocrinol Metab. 2001;86:4068.[br][br]Sources of Research Support: Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)-2005-04726-0 to ACL.[br][br]Nothing to Disclose: SD, VNB, PC, IJPA, MYN, BBM, ACL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1241 199 1634 SUN-622 PO29-01 Sunday 1431 2012


1428 ENDO12L_SUN-623 POSTER SESSION: Puberty (1:30 PM-3:30 PM) High Prevalence of Heterozygous CYP21A2 Carriers in Cypriot Girls with Premature Adrenarche Nicos Skordis, Christos Shammas, Alexia AP Phedonos, Meropi Toumba, Charilaos Stylianou, Elena Andreou, Michalis Picolos, Tassos C Kyriakides, Vassos Neocleous, Leonidas A Phylactou Makarios III Hospital, Nicosia, Cyprus; The Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus; Paphos General Hospital, Paphos, Cyprus; Makarios III Hospital, Nicosia, Cyprus; Alithias Center, Nicosia, Cyprus; Yale University, New Haven, CT Female carriers of [italic]CYP21A2[/italic] mutations are at increased risk of developing premature adrenarche (PA) during childhood which might evolve into polycystic ovary syndrome (PCOS) in adolescence. The present study was designed to seek evidence on the prevalence and consequences of heterozygous [italic]CYP21A2[/italic] mutations in girls with PA. The hormonal response to ACTH was evaluated in 81 girls with clinical signs of PA along with direct DNA sequencing and MLPA analysis for mutations in the [italic]CYP21A2[/italic] gene. The suspicion of heterozygote state was based on the median plasma 17-OHP before and 60 minutes after ACTH stimulation. Twelve girls were diagnosed with NC-CAH based on 17-OHP response and confirmed with genetic studies. Thirty five patients out of the 81 with PA were identified as carriers of [italic]CYP21A2[/italic] mutations. The most frequent mutations among the carriers were the mild p.V281L (54.3%), followed by p.Q318stop (20%), p.P453S (11.4%), p.V304M (5.7%), p.P482S (2.8%), delEX6-8 (2.8%) and the large deletion/conversion exons 1-4 (2.8%). Higher values of stimulated 17-OHP levels were found in the carriers of the p.V281L mutation compared with carriers of other mutations (Avg = 23.6 nmol/l [italic]vs[/italic] 16.1 nmol/l; [italic]P[/italic]=0.02). This finding supports the already identified notion that carriers of the mild p.V281L are at higher risk for hyperandrogenism than carriers of severe mutations. Additionally, a high allelic frequency of 61.8% of the p.N493S variant was observed in the group of 34 girls with PA and no mutation and which was significantly different when compared to the group of 35 heterozygous girls with PA (61.8% [italic]vs[/italic] 22.8%). In another study by our group that involved 300 Cypriot asymptomatic individuals the carrier frequency for p.N493S variant was estimated to be 37%. In conclusion: a. girls with PA are likely to bear heterozygous [italic]CYP21A2[/italic] mutations, therefore systematic evaluation of 17-OHP values in combination with the molecular testing of [italic]CYP21A[/italic]2 gene is beneficial, b. carriers of the mild p.V281L, are at higher risk of androgen excess compared to carriers of other types of mutations and c. hyperandrogenic signs in the group of girls with no mutation in [italic]CYP21A2[/italic] could implicate p.N493S variant as a plausible disease causing factor.[br][br]Nothing to Disclose: NS, CS, AAPP, MT, CS, EA, MP, TCK, VN, LAP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 221 199 1635 SUN-623 PO29-01 Sunday 1432 2012


1429 ENDO12L_SUN-624 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Is Intermediate Thelarche a New Entity Following a Different Clinical Course Than Premature Thelarche and Central Precocious Puberty? Carolina C Di Blasi, Christian Roth, Clinton T Elfers, Joyce P Yi-Frazier, Angela Badaru Seattle Children[apos]s Hospital and the University of Washington, Seattle, WA; Seattle Children[apos]s Research Institute, Seattle, WA; Seattle Children[apos]s Research Institute, University of Washington, Seattle, WA [bold]Introduction:[/bold] The spectrum of precocious sexual maturation is wide. Intermediate forms between benign premature thelarche (PT) and central precocious puberty (CPP) are common in practice. We describe an entity of [apos]Intermediate Thelarche[apos] (IT) as breast development in girls [lt]8yr who have features of CPP including pubic hair, and/or bone age advancement (adBA) but lack pubertal LH secretion (basal [ge]0.3 UsLH by ICMA or peak LH [ge]5mIU/mL after aq leuprolide).There are no recent studies examining outcomes in patients with IT or the utility of the current sensitive LH assays in IT. The aims of this study were to: describe according to etiology a large cohort of girls [lt]8yr referred for evaluation of precocious onset of puberty; compare baseline biochemical and clinical characteristics in patients with idiopathic CPP (iCPP), PT and IT and to describe the natural history of progression in girls with PT and IT (not iCPP due to intervention).[br][bold]Methods:[/bold] Retrospective chart review of 269 girls referred for PP 2006-2011, 42 were excluded due to incomplete data. 64 patients who met criteria for iCPP, PT or IT were analyzed for baseline and followup characteristics. iCPP was defined as B[ge]2, PH[ge]2 and/or adBA ([ge]2SDS) with pubertal LH and normal cranial MRI; PT as B[ge]2, no PH, no adBA and prepubertal LH; IT as B[ge]2, PH[ge]2 and/or adBA ([ge]2SDS) and prepubertal LH.[br][bold]Results:[/bold] Of the 64: 8 (12.5%) had iCPP, 17 (26.5%) PT and 39 (61%) IT. Of the remaining 163, 98 had precocious adrenarche, 30 prepubertal, 26 with brain pathology, 9 peripheral PP. At presentation, IT girls were similar in age to PT but younger than iCPP (Median: 6.66 vs 7.54yr, p[lt]0.05). Height and BMI z-scores were not different among groups, neither was the degree of adBA in IT and iCPP. Basal LH was higher in iCPP (2.5mIU/mL, p[lt]0.001) but similar in PT and IT (Mean: 0.05 and 0.03mIU/ml). Over the course of [ge]6 months, IT patients showed adBA in 17% of patients and pubertal LH values in 20% of patients.[br][bold]Conclusion:[/bold] Among girls who where referred for evaluation of precocious onset of puberty, we identified a group of patients that did not match criteria for iCPP nor for PT. We believe it is important to consider this group as a possible new entity, as they may have a risk to further progress into puberty, although their LH levels were prepubertal at first evaluation. Longitudinal prospective studies are needed to define outcomes in patients with this intermediate form of PP.[br][br]Nothing to Disclose: CCDB, CR, CTE, JPB-F, AB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1711 199 1636 SUN-624 PO29-01 Sunday 1433 2012


1430 ENDO12L_SUN-625 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Impact of Body Mass Index on Luteinizing Hormone Secretion in Gonadotropin-Releasing Hormone Stimulation Tests of Boys Experiencing Precocious Puberty Hae Sang Lee, Hong Kyu Park, Jung Hee Ko, You Jin Kim, Jin Soon Hwang Ajou University Hospital, Suwon, Korea Objective: Excess adiposity may influence various aspects of pubertal development, including the timing of pubertal initiation and hormonal parameters during puberty. The aim of the study was to evaluate the impact of obesity on luteinizing hormone (LH) secretion to gonadotropin-releasing hormone (GnRH) stimulation test in boys with precocious puberty.[br]Methods: Boys with precocious puberty, who were normal weight, overweight, and obese underwent GnRH-stimulation tests between 2003 and 2010. Subjects were classified as normal weight (BMI [ge]5[sup]th[/sup] percentile and BMI [lt] 85[sup]th[/sup] percentile), overweight (BMI [ge]85[sup]th[/sup] percentile and BMI [lt] 95[sup]th[/sup] percentile), and obese (BMI [ge]95[sup]th [/sup]percentile).[br]Results: Of 56 children whose data were included in the final analysis, mean age at diagnosis was 8.7 [plusmn] 1.0 years. The majority of boys were of normal weight (n=28, 50%), while 15 children (26%) were overweight, and 13 (23%) obese. Peak LH levels after GnRH stimulation were 19.8 [plusmn] 8.8, 9.0 [plusmn] 3.5, and 8.1 [plusmn] 4.0 mIU/mL among normal weight, overweight, and obese subjects, respectively ([italic]P[/italic][lt]0.001 for all comparisons). By multivariate analysis, there was a significant negative association of body mass index (BMI) with peak stimulated LH level.[br]Conclusion: The higher BMI is associated with lower LH response to the GnRH-stimulation test in boys experiencing precocious puberty. In boys with precocious puberty, BMI should be considered when interpreting GnRH-stimulation test.[br][br]Nothing to Disclose: HSL, HKP, JHK, YJK, JSH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 520 199 1637 SUN-625 PO29-01 Sunday 1434 2012


1431 ENDO12L_SUN-626 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Basal Luteinizing Hormone: Is It Sufficient for Diagnosis of Precocious Puberty in Korean Girls? Sun Hee Lee, Woo Yeoung Chung, Jae Hyun Kim Inje University College of Medicine, Busan Paik Hospital, Busan, Republic of Korea; Inje University College of Medicine, Ilsan Paik Hospital, Goyang, Republic of Korea Objective: A GnRH stimulation test (GnRHST) is a gold standard in diagnosing central precocious puberty. The aim of this study was to evaluate the auxological and biochemical characteristics of Korean girls who had undergone GnRHST and to investigate that basal LH could be a substitute method of GnRHST.[br]Methods: In 2 tertiary pediatric centers, Korean girls with early pubertal development were included during 2010-2011. Height, BMI, pubertal stage, chronological age and bone age were evaluated. LH, FSH, estradiol, IGF-I and IGFBP-3 were measured and a standard GnRHST was performed. The patients with peripheral precocious puberty were excluded. The peak LH level of [ge]5 IU/L was considered pubertal response during GnRHST.[br]Results: Among a total of 302 girls who had undergone GnRHST, 299 girls were included. Three were excluded because of peripheral precocious puberty caused by functional ovarian cysts. The chronological age (SDS) of 299 girls was 8.23 (0.68) year and bone age (SDS) was 9.90 (0.96) year at the time of a GnRHST. Height and BMI SDS was 0.86 (0.86) and 0.50 (1.00). The pubertal responses were observed in 226 girls (75.6%), and prepubertal responses were observed in 76 girls (24.4%). There were no significant differences in basal LH, peak LH, basal FSH, peak LH/FSH ratio, estradiol, height SDS and BMI SDS between these two groups. In logistic regression analysis, odds ratio of basal LH was 24.30 (95% C.I. 2.14-275.73, P=0.01). Receiver operating characteristic analyses showed that basal LH is a better predicting method for the pubertal result during GnRHST over basal FSH and estradiol. Area under the curve of basal LH, FSH and estradiol was 0.757, 0.714 and 0.565, respectively. Among 150 girls with a basal LH below the detection limit ([lt]0.1 IU/L), 89 (59.3%) had pubertal responses. There was no predicting factor in logistic regression analysis in girls with a basal LH below the detection limit.[br]Conclusion: An elevated level of basal LH was a significant predicting factor for diagnosis of central precocious puberty. However activation of hypothalamic-pituitary-gonadal axis was observed even in girls with a basal LH below the detection limit. GnRHST was still essential for a diagnostic confirmation of central precocious puberty.[br][br]Nothing to Disclose: SHL, WYC, JHK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 856 199 1638 SUN-626 PO29-01 Sunday 1435 2012


1432 ENDO12L_SUN-627 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Clinical Characteristics of Girls Presenting with Early Maturation Phil Soo Oh Hallym University Medical Center, Chuncheon, Korea [bold]Purpose[/bold]: The purpose of this study is to analyze the clinical and environmental characteristics of early maturated girls by physical examinations and questionnaire.[br][bold]Methods[/bold]: This study was done prospectively for 158 early maturated girls in the Department of Pediatrics, Hallym University Medical Center from July 2008 to June 2010.[br][bold]Results[/bold]: 1. For the 158 girls, the mean ages are 8.27[plusmn]1.3 years and mean bone age advancement is 1.86[plusmn]0.27 years. 2. In considerable cases, their weight-to-height percentiles is in 25[sim]50 and 50[sim]75. 3. They have normal/good balanced or meat-prone appetite in many cases (91%). 4. Family history of early maturation was about 40% positive. And in family relationship, the first girls have a tendency to be mature early twice more than in second. 5. Herb medicine taking history with ginseng was up to 67.5%.[br][bold]Conclusion[/bold]: 1. Unexpectedly, in considerable cases their weight-to-height percentiles is in 25[sim]50 or 50[sim]75. This result seems to be the possibility of any other cause for precocious puberty in addition to body fatness. 2. Interestingly in family relationship, the first girls have a tendency to be mature early twice more than in second. 3. In our study, herb medicine taking history with ginseng was up to 67.5%. With ginseng[apos]s phytoestrogenic effect in some reports, herb medicine seems to be an important factor for early maturation in Korea.[br][br]Nothing to Disclose: PSO 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 77 199 1639 SUN-627 PO29-01 Sunday 1436 2012


1433 ENDO12L_SUN-628 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Recovery of Normal Hypothalamic-Pituitary-Gonadal Function after Treatment with Subcutaneous Histrelin Implant for Central Precocious Puberty Lawrence A Silverman, Gad B Kletter, E Kirk Neely, Susan Potts, Errol Gould, Oksana Terleckyj, Theodore M Danoff Goryeb Children[apos]s Hospital, Morristown, NJ; Swedish Medical Center, Seattle, WA; Stanford University, Stanford, CA; Endo Pharmaceuticals, Inc, Chadds Ford, PA [bold]Introduction: [/bold]Gonadotropin-releasing hormone agonist (GnRHa) therapy is the standard of care for treatment of central precocious puberty (CPP). While a number of studies have reported long-term posttreatment follow-up data for injectable GnRHa treatment, there is limited long-term follow-up data for the once-yearly histrelin subcutaneous implant (Supprelin LA). An extended-access phase 3 open-label study demonstrated that up to 4 years of continuous histrelin implant therapy is effective in suppressing the hypothalamic-pituitary-gonadal (HPG) axis and improving auxologic outcomes in patients with CPP. We now report on the recovery of the HPG axis and auxologic outcomes in patients who returned for at least a 6-month posttreatment assessment after removal of their final implant. [bold]Methods: [/bold]GnRHa-stimulated peak luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels were measured 1 and 6 months after last implant removal. Basal estradiol levels in girls were measured at 6 and 12 months post-explant. Bone age/chronological age ratio and Bayley-Pinneau predicted adult height (PAH) were calculated 12 months post-explant. [bold]Results:[/bold] 8 children (7 girls; mean age at initial treatment 6.95 years [4.9-8.9 years]) who received a median of 5 implants (range, 3-6) returned for 6-month posttreatment assessments. Before the last implant was removed, mean stimulated LH was 0.30 mIU/mL and at 1 month post-explant increased to 3.62 mIU/mL ([italic]P[/italic]=.007). At 6 months post-explant, mean peak LH increased to 22.85 mIU/mL ([italic]P[/italic]=.020). Similarly, mean peak FSH increased from 1.82 mIU/mL at last implant removal to 6.23 mIU/mL ([italic]P[/italic]=.029) and 8.59 mIU/mL ([italic]P[/italic][lt].001) within 1 and 6 months post-explant, respectively. Among the 7 girls, mean basal estradiol increased significantly from 5.50 pg/mL prior to removal of last implant to 23.84 pg/mL at 6 months ([italic]P[/italic]=.002) and 29.67 pg/mL at 12 months post-explant ([italic]P[/italic][lt].001). Mean bone age/chronological age ratio was significantly lower 12 months post-explant compared with baseline (1.13 vs 1.37; [italic]P[/italic]=.001) and time of last implant removal (1.13 vs 1.15; [italic]P[/italic]=.011), leading to an 8-cm increase in mean PAH at 12 months post-explant compared with baseline (162.22 cm vs 154.21 cm; [italic]P[/italic]=.017). No SAEs were reported. [bold]Conclusions:[/bold] After discontinuation of up to 6 years of histrelin therapy, swift recovery of the HPG axis occurs. Histrelin therapy also resulted in a significant increase in PAH in girls treated for a median of 5 years.[br][br]Sources of Research Support: Endo Pharmaceuticals, Inc.[br][br]Disclosures: LAS: Researcher, Endo Pharmaceuticals Inc., Pfizer, Inc.; Speaker Bureau Member, Endo Pharmaceuticals Inc.; Consultant, Endo Pharmaceuticals Inc., Pfizer, Inc., Novo Nordisk. EKN: Researcher, Endo Pharmaceuticals Inc., Abbott Laboratories. SP: Employee, Endo Pharmaceuticals Inc. EG: Employee, Endo Pharmaceuticals Inc. OT: Employee, Endo Pharmaceuticals Inc. TMD: Employee, Endo Pharmaceuticals Inc. Nothing to Disclose: GBK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1781 199 1640 SUN-628 PO29-01 Sunday 1437 2012


1434 ENDO12L_SUN-629 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Clinical Efficacy of Three Month Depot GnRH Agonist in Suppression of Central Puberty Mandi L Cafasso, Deborah A Elder, Samantha D Blum, Tammy L Weis, Barry E Vogel, Susan R Rose Cincinnati Children[apos]s Hospital Medical Center and University of Cincinnati, Cincinnati, OH [bold]Objective[/bold]: Precocious puberty, and puberty occurring at inappropriately short height, are treated with GnRH agonist (GnRHa) to suppress puberty.Monthly GnRHa is FDA-approved for pediatric use. A longer duration (3 month) GnRHa has been FDA-approved for suppression of precocious puberty. Features of pubertal suppression from administration of monthly GnRHa were compared to features with the 3m GnRHa. We hypothesized that suppression from the two GnRHa forms would be equivalent. We extend observations of 3m GnRHa to longer duration of follow up.[br][bold]Methods[/bold]: IRB-approved retrospective chart review was performed of data from 79 patients (58 female) at baseline, 1-6m, and yearly follow-up of GnRHa therapy, including height, weight, BMI, growth velocity, bone age and serum hormone levels.[br][bold]Results[/bold]: 45 patients were treated with monthly GnRHa and 34 treated with 3m GnRHa. There was no difference between the two groups in duration of therapy (3.0 vs 2.7y), slowing of BA advance (0.8 vs 0.7y), yearly BMI increase (1.0 vs 1.0SD), growth velocity on therapy (6.6 vs 6.2cm), or years of height age gain compared to change in BA (1.4 vs 1.3y). At start of treatment, age was younger (7.3 vs 9.7y) and BA advance (2.3 vs 1.3y) was greater in monthly group. Pubertal suppression was not deemed adequate in 15 (36%) of monthly group, requiring increased frequency of GnRHa injection in 6 patients to every 3 weeks, and requiring change to 3m GnRHa in 9 patients. Suppression was not adequate in 3 (8%) of 3m group, requiring increased frequency to every 9-10 weeks. An additional 12 (28%) of monthly group switched to 3m GnRHa for convenience or to reduce stress from shots. None switched from 3m to monthly agonist.[br][bold]Conclusions[/bold] [ndash] Clinically, 3m GnRHa was able to suppress puberty in 92% by time of 2[sup]nd[/sup] injection, thus was at least as effective as suppression achieved by standard monthly GnRHa (79%). Puberty in most patients without suppression could be controlled with agonist therapy, if doses were moved closer together. Features such as accelerated growth rate, pre-injection moodiness and physical examination features (breast development, testicular enlargement) were clinical indications to adjust frequency of administration in a small number of patients using either medication form. Suppression of puberty with 3m GnRHa is as efficacious as standard monthly GnRHa treatment, and should be considered in patients with precocious puberty or puberty occurring at an inappropriately short height.[br][br]Nothing to Disclose: MLC, DAE, SDB, TLW, BEV, SRR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 647 199 1641 SUN-629 PO29-01 Sunday 1438 2012


1435 ENDO12L_SUN-630 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Histrelin Subdermal Implants Provide Continuous Suppression of the Hypothalamic-Pituitary-Gonadal Axis for 4 Years in Children with Central Precocious Puberty Lawrence A Silverman, Gad B Kletter, E Kirk Neely, Susan Potts, Errol Gould, Oksana Terleckyj, Theodore M Danoff Goryeb Children[apos]s Hospital, Morristown, NJ; Swedish Medical Center, Seattle, WA; Stanford University, Stanford, CA; Endo Pharmaceuticals, Inc, Chadds Ford, PA [bold]Introduction:[/bold] Children with central precocious puberty (CPP) often require several years of gonadotropin-releasing hormone agonist (GnRHa) therapy. Continuous suppression of the hypothalamic-pituitary-gonadal (HPG) axis with some GnRHa therapies requires intramuscular injections that can be painful and may impede patient adherence. A once-yearly subcutaneous histrelin implant (Supprelin LA) eliminates the need for frequent intramuscular injections. Results from an extended-access open-label phase 3 study demonstrated that sequential 12-month histrelin implants suppress the HPG axis through 3 years. We now report the results of the long-term extension stage of the phase 3 trial, which evaluated the efficacy and safety of 12-month histrelin implant therapy for a fourth year. [bold]Methods:[/bold] Patients who received a fourth implant are included in this analysis. [bold]Results:[/bold] Thirteen children (12 girls; mean age at baseline [time of first implant] 6.6 years [range, 4.5-9.1 years]) received a fourth implant. Among 8 girls who were GnRHa-treatment na[iuml]ve at initial enrollment, GnRHa-stimulated mean peak LH levels at 48 months were significantly suppressed compared with baseline (0.33 mIU/mL vs 20.44 mIU/mL; [italic]P[/italic]=.016). Mean peak FSH levels at 48 months were also significantly suppressed compared with baseline (1.89 mIU/mL vs 15.76 mIU/mL; [italic]P[/italic]=.005). Among the 5 patients previously treated with a GnRHa prior to initial enrollment, mean peak LH was 2.95 mIU/mL at baseline, 0.66 mIU/mL ([italic]P[/italic]=.090) at 1 month, and remained suppressed at 48 months (0.40 mIU/mL). Among all the girls, mean estradiol level at 48 months was 5.22 pg/mL (vs 6.58 pg/mL at baseline; [italic]P[/italic]=.285). For the boy, testosterone level was 9.9 ng/dL at baseline and 7.3 ng/dL at 48 months. Among all the patients, mean bone age/chronological age ratio was significantly lower (1.16 vs 1.41; [italic]P[/italic][lt].001) and Bayley-Pinneau predicted adult height was significantly higher (157.36 cm vs 150.05 cm; [italic]P=[/italic].015) at 48 months compared with baseline. During 4 years, the most frequently reported adverse event was implant site reaction (pain and discomfort) occurring in 9 patients. No patient discontinued during the fourth year of histrelin implant therapy. [bold]Conclusion:[/bold] Four years of continuous histrelin implant therapy is effective in suppressing the HPG axis and improving auxologic outcomes in patients with CPP while eliminating the need for intramuscular injections associated with other GnRHa therapies.[br][br]Sources of Research Support: Endo Pharmaceuticals, Inc.[br][br]Disclosures: LAS: Researcher, Endo Pharmaceuticals Inc., Pfizer, Inc.; Speaker Bureau Member, Endo Pharmaceuticals Inc.; Consultant, Endo Pharmaceuticals Inc., Pfizer, Inc., Novo Nordisk. EKN: Researcher, Endo Pharmaceuticals Inc., Abbott Laboratories. SP: Employee, Endo Pharmaceuticals Inc. EG: Employee, Endo Pharmaceuticals Inc. OT: Employee, Endo Pharmaceuticals Inc. TMD: Employee, Endo Pharmaceuticals Inc. Nothing to Disclose: GBK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1695 199 1642 SUN-630 PO29-01 Sunday 1439 2012


1436 ENDO12L_SUN-631 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Pharmacokinetic and Exposure-Response Analysis of Leuprolide Following Administration of Leuprolide Acetate 3-Month Depot Formulations to Children with Central Precocious Puberty Nael Mostafa, Balakrishna Hosmane, Lois Larsen, Kristof Chwalisz, Yi-Lin Chiu, Rajendra Pradhan Abbott Laboratories, Abbott Park, IL; Northern Illinois University, DeKalb, IL [bold]BACKGROUND: [/bold]Leuprolide acetate (LA) is a synthetic gonadotropin-releasing hormone agonist (GnRHa) approved for treatment of children with central precocious puberty (CPP).[br][bold]OBJECTIVE: [/bold]Characterizing leuprolide (L) pharmacokinetics for 2 LA depot formulations in children with CPP and assessing the exposure-response (ER) relationship between L concentrations and the probability of luteinizing hormone (LH) suppression.[br][bold]DESIGN/METHODS: [/bold]A Phase 3 study was conducted in 84 children with CPP and evaluated 2 formulations (LA Depot 11.25 mg and 30 mg) in either subjects previously treated with GnRHa (N = 21/42) or in GnRHa-na[iuml]ve subjects (N = 21/42) following intramuscular injections of LA every 3 months for 6 months (2 injections). Serial L concentrations were measured in a subset of 48 children. Linear mixed effects modeling and trend analysis were used to assess attainment of steady state and LA accumulation. One-way analysis of covariance was used to assess dose proportionality. The probability of LH suppression (peak-stimulated LH concentrations [lt] 4 mIU/mL) ER relationship utilized repeated measures logistic regression accounting for correlation among multiple time points from a subject. The predicted probability of LH suppression and corresponding 95% confidence interval (CI) at the mean L concentration of each dose group and at each time of measurement (4, 8, 12, and 24 weeks [wks]) were computed.[br][bold]RESULTS:[/bold] The mean L concentration remained constant from Wk 4 to 12 for both the 11.25 and 30 mg doses ([italic]P [/italic][gt] 0.29). Mean 12-wk L concentrations after the first and second injections were similar for both dose groups ([italic]P [/italic][gt] 0.51). L concentrations from Wk 4 to 12 were dose proportional across the 11.25 mg and 30 mg doses ([italic]P [/italic][gt] 0.64). The effects for age ([italic]P [/italic][gt] 0.62) and weight ([italic]P [/italic][gt] 0.27) were not statistically significant. The final ER model was:[br]Log [p/(1 [ndash] p)] = [alpha] + [beta][bull]Baseline + [nu][bull]Log (concentration) + [lambda][bull]Wk[br]where [alpha], [beta], [nu], and [lambda] are logistic regression coefficients. The model showed that, across all time points, higher L concentrations were associated with higher probability of LH suppression ([italic]P [/italic]= 0.02).[br][bold]CONCLUSIONS:[/bold] Mean L concentrations between Wks 4 and 12 for both 11.25 and 30 mg doses were relatively constant and dose-proportional, with no accumulation of L upon repeated administration. Although higher L concentrations were associated with a higher probability of LH suppression, both doses provided mean LH suppression levels [lt] 4 mIU/mL.[br][br]Disclosures: NM: Employee, Abbott Laboratories. BH: Consultant, Abbott Laboratories. LL: Researcher, Abbott Laboratories. KC: Employee, Abbott Laboratories. Y-LC: Employee, Abbott Laboratories. RP: Employee, Abbott Laboratories. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 367 199 1643 SUN-631 PO29-01 Sunday 1440 2012


1437 ENDO12L_SUN-632 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Near Adult Height and Time of Menarche of Girls with Idiopathic Central Precocious Puberty and Early Puberty after Gonadotropin-Releasing Hormone Agonist Treatment Hong Kyu Park, Hae Sang Lee, Jung Hee Ko, You Jin Kim, Jin Soon Hwang Ajou University Hospital, Suwon, Korea [bold]Objectives[/bold][br]Gonadotropin-releasing hormone (GnRH) agonist has been used for the treatment of girls with central precocious puberty (CPP) and early puberty (EP) for more than two decades. Effect on final height outcome is uncertain yet.[br][bold]Methods[/bold][br]A prospective analysis of 39 patients (6 with CPP and 39 with EP) was assessed to evaluate the correlation between predicted adult height and near final height.[br][bold]Results[/bold][br]The mean age of the subjects at the start of treatment was 9.1 [plusmn] 0.8 years old (8.4 [plusmn] 0.3 in CPP, 9.3 [plusmn] 0.8 in EP). Bone age was 10.3 [plusmn] 0.8 years old (10.1 [plusmn] 0.7 in CPP, 10.7 [plusmn] 0.8 in EP) and was advanced by 1.5 [plusmn] 0.5 years. Standard deviation score (SDS) of height was 0.63 [plusmn] 0.66 before the treatment. Predicted adult height (PAH) was 153.0 [plusmn] 5.7 cm, and it was 1.60 [plusmn] 1.20 standard deviation below the mean of young adult women. The mean duration of treatment was 2.1 [plusmn] 0.6 years. PAH at the end of treatment was increased to 157.7 [plusmn] 5.6 cm (-0.63 [plusmn] 1.14 SDS). The mean of height gain from the discontinuation of GnRH agonist was 13.9 cm (7.72 [ndash] 25.3 cm) and menarche occurred 17 months (3 [ndash] 36 months) after discontinuation of treatment. The final height (FH) was 159.3 [plusmn] 4.6 cm (-0.3 [plusmn] 0.9 SDS) and was 2.2 [plusmn] 3.7 cm taller than mid-parental height. FH was 5.9 [plusmn] 3.3 cm higher than the initial PAH, and 1.2 [plusmn] 2.7 cm higher than the PAH at the discontinuation. The Pearson correlation of PAH and FH was 0.809 (p [lt] 0.001).[br]Five of 39 patients (12.8%) were received combined treatment with human recombinant growth hormone. The mean duration was 12 months. No statistical difference was observed between the PAH, FH, or duration to menarche, and growth hormone administration.[br][bold]Conclusions[/bold][br]GnRH agonist administration in girls with CPP and EP demonstrated an improvement in final height outcome over PAH based on the Bayley-Pinneau method.[br][br]Nothing to Disclose: HKP, HSL, JHK, YJK, JSH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1042 199 1644 SUN-632 PO29-01 Sunday 1441 2012


1438 ENDO12L_SUN-633 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Histrelin Implant for Precocious Puberty [mdash] Final Height and Time to Menarche Harry J Hirsch, David Strich, Gilad Karavani, David Gillis Shaare Zedek Medical Center, Jerusalem, Israel; Clalit Health Services, Jerusalem, Israel; Hadassah-Hebrew University Medical School, Jerusalem, Israel; Hadassah-Hebrew University Medical Center, Jerusalem, Israel [bold]Background:[/bold] The subcutaneous histrelin implant (HI) has been shown to suppress gonadotropin secretion in children with central precocious puberty (CPP) and effectively treat CPP for 12-months or more. Treatment of CPP with the HI results in greater suppression of LH and FSH compared to treatment with depot GnRHa[sup]1[/sup]. Following removal of the implant, free alpha-subunit levels decrease within one week, serum LH and FSH rise after three weeks, and estradiol increases 6 weeks after removal[sup]2[/sup]. It is not known if the rapid recovery from gonadotropin suppression results in a shorter time until menarche or if the final heights in children treated with the HI are the same as those treated with depot GnRHa.[br][bold]Objective:[/bold] To compare final height and time from end of treatment until menarche in girls with CPP treated with the HI versus depot-triptorelin injections.[br][bold]Method:[/bold] Chart review, interview and final height measurements of two groups of girls with CPP; group A (implant group) included 8 girls treated from age 8.2[plusmn]0.5 years. They initially received monthly injections of triptorelin and were switched to the HI after 9.6[plusmn]4.1 months for the remainder of treatment. Group B included 14 girls treated from age 8.1[plusmn]0.8 years with monthly injections of triptorelin for the entire treatment period. Final height and the time between either implant removal (group A) or last injection (group B) to menarche were assessed and compared with a two tailed t-test for groups with unequal variance. Total duration of treatment for group A was 40.2[plusmn]4.9 months vs. 28.1[plusmn]3.5 months for group B (p=0.06).[br][bold]Results:[/bold] Average age at menarche in group A was 13.0[plusmn]0.3 years vs. 12.4 years in group B (NS). Average time between termination of treatment and menarche was 8.9[plusmn]1.5 months in group A vs.17.2[plusmn]2.5 months for group B (p[lt]0.01). Final height was 0.7[plusmn]2.2 cm. above midparental height for group A vs. 3.0[plusmn]2.5 cm. below midparental height for group B (NS). Adult height prediction by bone age at implant insertion was 164.5[plusmn]2.3 cm. and final height was 161.34[plusmn]2.69 cm. Similarly predicted height for triptorelin patients was 160.7[plusmn]2.5 cm. and final height was 157.48[plusmn]2.5 cm. (NS).[br][bold]Conclusions:[/bold] Menstruation occurs sooner after removal of the HI compared with after cessation of depot-triptorelin injections, but final height gain is similar for the two groups. The observed trend towards taller final height relative to parental height in group A may be due to a longer treatment period in the implant group.[br][br]1. Hirsch et al., Pediatrics 2005; 116: e798. 2. Hirsch et al., J Clin Endocrinol Metab 2010; 95: 2841.[br][br]Sources of Research Support: Endo pharmaceuticals.[br][br]Nothing to Disclose: HJH, DS, GK, DG 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2074 199 1645 SUN-633 PO29-01 Sunday 1442 2012


1439 ENDO12L_SUN-634 POSTER SESSION: Puberty (1:30 PM-3:30 PM) Body Mass Index, Rate of Obesity, Fasting Glucose and Lipid Profiles in Children with Central Precocious Puberty and Early Puberty before and during Either Three Years of Treatment with a GnRH Analog or Follow-Up without Therapy Ana Colmenares, Peter Gunzler, Roberto Lanes Instituto Venezolano del Seguro Social, San Cristobal, Venezuela; Hospital de Cl[iacute]nicas Caracas, Caracas, Venezuela Aims: To assess the changes in body mass index (BMI), obesity rate, fasting glucose and lipids in children with CPP and EP treated with a GnRHa for 3 years and to compare these results to those of a similar group of non-treated patients.[br]Methods: We analyzed the data of 100 patients with CPP/EP (89 girls); 56 were treated with a GnRHa for 3 yrs for rapidly progressing puberty, while 44 were followed for 3 years without treatment with slowly progressing puberty. Anthropometric parameters, percentage of obesity, glucose and lipids were similar in the 2 groups at baseline. Data were evaluated before, during and after 3 years of treatment or of follow up. Obesity was defined +2 SDS using Venezuelan reference data for BMI.[br]Results: At the time of diagnosis, patients with CPP/EP had an elevated BMI SDS (2.5[plusmn]2.1 in GnRHa treated and 2.5[plusmn]2.1 in non treated subjects, p=0.8) and a high rate of obesity (55% and 52% in GnRHa treated and non-treated patients, p= 0.8). In GnRHa-treated patients mean BMI SDS at 2 years remained unchanged, but a significant increase was detected during the third year (p[lt]0.04). There was, however, no difference between BMI SDS before therapy and at the end of therapy (p=0.07). BMI SDS was greater than +2 SDS in 55% of patients after 1 year, 56.5% at 2 years, and 62.5% after 3 years of therapy (NS). In non-treated patients, BMI SDS and the obesity rate decreased during follow up, although not significantly (BMI SDS of 2.5[plusmn]2.1 at baseline vs.1.9[plusmn]1.9 at 36 months, p=0.2; obesity rate of 52% at baseline vs. 29% at 36 months, p=0.07). At the end of 3 years, the prevalence of obesity was significantly higher in GnRHa-treated than in untreated subjects (62.5% vs. 29%, p[lt]0.04), while no statistical difference in BMI SDS was noted between groups (BMI SDS of 2.3[plusmn]1.7 vs. 2.3 [plusmn]1.9 at 12 months, p=0.7; 2.5[plusmn]1.8 vs. 2.2[plusmn]1.7 at 24 months, p=0.2 and 2.5[plusmn]2.1 vs.1.9[plusmn]1.9 at 36 months, p=0.2). Mean fasting glucose, total and LDL cholesterol and triglycerides remained unchanged during the 3 years in both groups.[br]Conclusions: The rate of obesity among children with CPP/EP prior to therapy is elevated. The percentage of patients with a BMI SDS [gt]+2 is significantly higher in GnRHa treated vs. untreated patients after 3 years. However, BMI SDS of GnRHa treated patients did not increase with therapy. Fasting glucose and lipids remained stable in all patients during the 3 year period.[br][br]Nothing to Disclose: AC, PG, RL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1000 199 1646 SUN-634 PO29-01 Sunday 1443 2012


1440 ENDO12L_SUN-664 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Proteomic Analysis of Brain Tissue from Transgenic bGH and Wild-Type Mice Amrita Basu, Elahu Gosney, Edward O List, John Kopchick Ohio University, Athens, OH; Ohio University, Athens, OH; Ohio University Heritage College of Osteopathic Medicine, Ohio University, Athens, OH; Ohio University, Athens, OH Experimental evidence of GH and GHR presence in different regions of the brain (e.g. hippocampus, amygdale, and choroid plexus) indicates an influence of GH signaling on the structure and function of the central nervous system. Extremes in GH signaling, as observed in acromegaly (increased GH signaling), dwarfism (reduced GH signaling) or Laron Syndrome (absence of GH signaling due to dysfunctional GHR) in human patients has also been correlated with modification in neuronal cell proliferation, myelination, cognitive performance, memory processing and general mood and behavior. The detailed molecular mechanism underlying the function of GH in the brain is not yet well understood. Since the effect of GH may be mediated by altered protein expression in the brain, proteomic analysis of brain tissue obtained from mouse lines with altered GH action would bring a greater understanding to the role of GH action in the brain. In order to estimate the variation of protein expression due to altered GH signaling, total protein was isolated from whole brain of 12 week old male bovine GH (bGH) transgenic mice and normal wild type controls. Two dimensional gel electrophoresis of the protein samples (n=5) was followed by staining with SyproOrange[sup]TM [/sup]and analysis of the observed protein [apos]spots[apos] with PDQuest[sup]TM[/sup] software. Differential expression of protein was observed with significantly reduced levels in bGH mouse brain compared to wild type. Further analysis including mass spectrometry, functional characterization of proteins of interest and characterization of brain proteome from other mouse lines will help to elucidate the role of GH in the central nervous system and could help identify novel therapeutic targets.[br][br]Sources of Research Support: This work was supported in part by the State of Ohio[apos]s Eminent Scholar Program that includes a gift from Milton and Lawrence Goll, by the AMVETS, by the Diabetes Research Initiative at Ohio University, and by NIH grants DK083729, AG031736.[br][br]Nothing to Disclose: AB, EG, EOL, JK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2061 200 1647 SUN-664 PO18-01 Sunday 1444 2012


1441 ENDO12L_SUN-665 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) The Role of Zinc Binding and the Cellular Zinc Content in Regulated Growth Hormone Secretion Vibor Petkovic, Maria Consolata Miletta, Andree Eble, Daniel Iliev, Gerhard Binder, Christa Emma Fluck, Primus Eugen Mullis University Children[apos]s Hospital, Bern, Switzerland; University Children[apos]s Hospital, T[uuml]bingen, Germany The presence of zinc (Zn[sup] 2+[/sup]) in human pituitary growth hormone (GH)-secretory granules in equimolar amounts with GH is prerequisite for the formation of soluble dimer complexes consisting of two GH molecules and two Zn[sup] 2+[/sup] ions. Zn[sup] 2+[/sup] binding by GH through amino acid residues His18, His21, Glu174 seems to be essential for proper GH dimerization and its condensation and storage in secretory granules. Hence, [italic]GH-1[/italic] gene mutations at these positions might interfere with these processes having a final impact on normal GH secretion.[br]To investigate this more in detail, [italic]wt[/italic]-GH and/or triple-Zn[sup] 2+[/sup] GH mutant (GH-H18A-H21A-E174A) were transiently expressed in GC-cells (rat pituitary cell line) and extracellular GH secretion and intracellular GH production were analyzed in forskolin-stimulated vs. non-stimulated conditions in complete growth medium and medium treated with TPEN, the high-affinity Zn[sup] 2+[/sup] chelator.[br]Our secretion studies revealed no difference in constitutive GH secretion (without stimulation) of triple-Zn[sup] 2+[/sup] GH mutant compared to the [italic]wt[/italic]-GH while in forskolin-stimulated conditions, after 24h of stimulation, the mutant GH displayed significantly reduced extracellular secretion when compared to [italic]wt[/italic]-GH. Moreover, these differences in stimulated-secretion were even more pronounced when free Zn[sup] 2+[/sup] was depleted from culture medium. In general, stimulated GH secretion in complete culture medium was significantly higher when compared to that in TPEN-treated medium confirming important role of free Zn[sup] 2+[/sup] in the process of GH secretion.[br]Intracellular GH production, analyzed by western blot, revealed lower intracellular expression of [italic]wt[/italic]-GH when compared to that of triple-Zn[sup] 2+[/sup] GH mutant in non-stimulated and forskolin-stimulated conditions. Of note is that apoptosis studies analyzed through annexin-V expression in cells cultured in complete vs. TPEN-treated medium, confirmed non-toxic effect of TPEN.[br]Taken together, the data of GH secretion/production support the hypothesis that not only the loss of the affinity of mutant GH to Zn[sup] 2+[/sup] but also the cellular free Zn[sup] 2+[/sup] content as such could interfere with the normal GH dimerization and storage of triple-Zn[sup] 2+[/sup] GH mutant alone and with [italic]wt[/italic]-GH and could possibly explain impaired GH secretion.[br][br]Nothing to Disclose: VP, MCM, AE, DI, GB, CEF, PEM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 745 200 1648 SUN-665 PO18-01 Sunday 1445 2012


1442 ENDO12L_SUN-666 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Pericytes: Multitask Cells in Pituitary Functioning Tatiana Fiordelisio, Chrystel Lafont, Pierre-Francois Mery, Patrick Samper, Pierre Fontanaud, Jeremy Deverdun, Francois Molino, Anne Guillou, Agnes Martin, Oliver Mai, Ulrich Boehm, Karine Rizzoti, Robin Lovell-Badge, Patrice Mollard Institute of Functional Genomics, Montpellier, France; Universidad Nacional Aut[oacute]noma de M[eacute]xico, Mexico City, Mexico; Institute for Neural Signal Transduction, Hamburg, Germany; NIMR/MRC, London, UK Pericytes are contractile mural cells, which line blood microvessels. In the past two years, studies have also highlighted an important contribution of pericytes to the tissue micro-environment, including blood-brain-barrier integrity and permeability, trans-vascular migration of immune cells and even spinal cord repair. Adding to this growing list, the present study suggests that neural crest(NC)-derived pericytes are also key players in the micro-environment and function of pituitary cell networks (1-3). Together with the emerging microvasculature, NC-derived pericytes start to invade the pituitary parenchyma from E14.5 (Wnt1Cre/Sox10Cre mice). In adult mice, NC-derived pituitary pericytes are then located both along the capillaries and within the parenchyma, thus making anatomical bridges between capillaries. NC-derived pericytes are also organized as a rostro-caudal gradient on the ventral side of the pituitary. Pericyte functioning was explored by using both cellular in vivo imaging (4) and in situ multi-cellular imaging (1, 5). First, NC-derived pericytes can control blood vessel dynamics in the presence and absence of secretagogue (GHRH) stimulation. Second, they are involved in the compartmentalization of secreted hormones from their releasing sites towards fenestrated capillaries. Third, they contribute to the build-up of GH pulses in response to GHRH stimulation. Taken together, these data suggest that pericytes are multitask cells of the pituitary microenvironment. In addition to their role on capillary blood flow and thereby oxygen/nutrient supply of endocrine cell networks, they participate to the clearance of hormones within the pituitary. More broadly, studies of pericyte functioning might provide new and potentially unexpected insights into the pituitary microenvironment in both normal and pathological conditions.[br][br](1) Sanchez-Cardenas C et al. PNAS 2010; 107:21878. (2) Budry L et al. PNAS 2011; 108:12515. (3) Hodson DJ et al. Nat Commun. 2012; doi: 10.1038/ncomms1612. (4) Lafont C et al. PNAS 2010, 107:4465. (5) Schaeffer M et al. Endocrinology 2011; 152:4789.[br][br]Sources of Research Support: ANR grant [quot]Opto-Rhythms[quot] awarded to PM.[br][br]Nothing to Disclose: TF, CL, P-FM, PS, PF, JD, FM, AG, AM, OM, UB, KR, RL-B, PM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1918 200 1649 SUN-666 PO18-01 Sunday 1446 2012


1443 ENDO12L_SUN-667 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) GH Peak Amplitude Is Transiently Blunted in Young Preproghrelin Deficient Mice Rim Hassouna, Philippe Zizzari, Catherine Tomasetto, Jacques Epelbaum, Virginie Tolle INSERM, Universit[eacute] Paris Descartes, Paris, France; Institut de G[eacute]n[eacute]tique et de Biologie Mol[eacute]culaire et Cellulaire (IGBMC), Universit[eacute] Louis Pasteur, Illkirch, France Ghrelin is a gut hormone originating from the post translational cleavage of preproghrelin and is the endogenous ligand of the Growth Hormone Secretagogue Receptor 1a (GHSR-1a). However, the role of the endogenous peptide in the regulation of pulsatile Growth Hormone (GH) secretion remains to be established. The main phenotypic change observed so far in preproghrelin Knock-Out (ghrl -/-) mice is resistance to diet induced obesity when started shortly after weaning but not when started in older animals. The aim of this study was thus to compare the role of endogenous ghrelin in GH secretion at 7 weeks and 8 months of age in ghrl -/- mice. In order to assess GH pulsatile secretion, blood samples were collected by tail bleeding every 10 min over a period of 6 h in unaesthetized ghrl -/- mice and wild type (WT) littermates. Cluster analysis showed a pulsatile GH secretory pattern in all genotypes at both ages. However GH peaks amplitude was significantly reduced in young ghrl -/- mice compared to the WT littermates while at the age of 8 months there was no difference between genotypes. These results may suggest that ghrelin is a transient regulator of GH peak amplitude. The origin of the reduction in the GH pulsatile secretion in young ghrl -/- mice remains to be determined. A delayed growth or a greater sensitivity of the GH/IGF-1 axis to stress can be hypothesized.[br][br]Sources of Research Support: INSERM and Paris Descartes University.[br][br]Nothing to Disclose: RH, PZ, CT, JE, VT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1890 200 1650 SUN-667 PO18-01 Sunday 1447 2012


1444 ENDO12L_SUN-668 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Growth Hormone Sensitivity Varies Diurnally in FVB Mice Philip A Berry, Yue Zhang, Jing Jiang, Martin E Young, Stuart J Frank University of Alabama at Birmingham School of Medicine, Birmingham, AL; University of Alabama at Birmingham School of Medicine, Birmingham, AL; University of Alabama at Birmingham School of Medicine, Birmingham, AL; Birmingham VAMC, Birmingham, AL Important aspects of metabolism follow daily oscillations. Environmental cues such as light and temperature govern this rhythmicity. Although circadian rhythms have long been observed, their molecular basis and purpose are only now being unveiled. The circadian clock has two tiers; the upper tier is a CNS-located central clock that receives cues from the entire organism[apos]s environment, whereas the lower tier consists of transcription-translation feedback loops (the cell autonomous clock) present in nearly every cell in the body. The putative function of this system is to coordinate fluctuating metabolism of an individual cell with larger changes in the organism[apos]s environment such as temperature and light. Growth hormone (GH) exhibits both pulsatile and diurnal variation in humans, with approximately eight pulses per day and the greatest amplitude pulses in the night. Studies in rodents indicate similar pulsatility of circulating GH, except that in the nocturnal rodent, the rhythm for circulating GH is opposite that in humans in that the highest values occur during the day. Although the pulsatility and diurnal variation of GH secretion is well documented, little is known about potential diurnal variation in growth hormone sensitivity of target tissues. As GH action is central to the regulation of fuel metabolism and circadian rhythms are intimately related to metabolism, we began studies in male FVB mice to examine if growth hormone receptor (GHR) abundance and GH sensitivity vary diurnally. 18-22wk old male FVB mice were injected IP with GH (0.2mcg/g or 0.05mcg/g) vs saline at 12 noon (ZT6) vs 12 midnight (ZT18), then sacrificed after 15 minutes. Immunoblots of liver, heart, skeletal muscle, and adipose tissue lysates, but not kidney lysates, revealed greatly increase GH-dependent STAT5 phosphorylation (normalized by total STAT5 level) at ZT6 compared to ZT18. Hepatic GHR protein abundance was also elevated at ZT6 vs ZT12, whereas other tissues exhibited unchanged GHR abundance. Mechanisms underlying these phenomena are being investigated but are currently unclear, with time of day oscillations at one of several levels in the proximal GHR signal cascade potentially accounting for altered STAT5 activation: altered GHR expression at mRNA level, GHR turnover via [alpha]-secretase-mediated cleavage or endocytosis, phosphatase activity on JAK2 or GHR by SHP-2 or PTP1B, adaptor proteins increasing JAK2 activity such as SH2-B, or inhibition via SOCS/CIS.[br][br]Sources of Research Support: VA Merit Review Award (SJF).[br][br]Nothing to Disclose: PAB, YZ, JJ, MEY, SJF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1359 200 1651 SUN-668 PO18-01 Sunday 1448 2012


1445 ENDO12L_SUN-669 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) The Targeted Secretion Inhibitor (TSI) SXN101959 Produces Dose-Dependent Inhibition of Pulsatile Growth Hormone Secretion in Male Rats after Intravenous Administration Elaine A Harper, Alberto Martinez, Anne Wilson, Sandra Marlin, Julien Browne Syntaxin Ltd, Abingdon, UK Previous studies have shown that SXN101742 (a histidine tagged version of SXN101959) with a modified growth hormone releasing hormone targeting domain and endopeptidase domain of botulinum serotype D, produces an inhibition of GH secretion following intravenous (i.v.) administration to male rats. Here we investigate the nature and extent of the inhibition of GH secretion produced by SXN101959 after i.v. dosing in male rats. Two separate studies were performed. In the first study, twenty-eight femoral vein and artery cannulated Sprague Dawley rats, four per treatment group, received either vehicle (50mM HEPES pH 7.2, 150mM NaCl) or SXN101959 (0.001, 0.003, 0.01, 0.03, 0.1 or 0.3 mg/kg, i.v.). In the second study, forty rats, eight per group, received either vehicle, 0.01, 0.03, 0.1 or 0.3mg/kg SXN101959 (i.v.). Rats were dosed on day 1 and on day 5 bloods were collected via an automated blood sampling system every 15 min from 19:00 through to 05:45. Plasma GH levels were determined by PerkinElmer AlphaLISA.[br]In both studies, vehicle treated rats showed pulses of GH secretion every three hours with peak maxima in the range of [sim]60 - [sim]140ng/ml. In addition, in both studies SXN101959 produced a significant dose-dependent decrease in pulsatile GH secretion (p[lt]0.005, one way ANOVA). Thus, in rats treated with SXN101959 doses of up to 0.03 mg/kg the profile number and size of GH pulses although variable was reduced. In rats treated with 0.1 mg/kg both the number of pulses and peak maxima were clearly reduced and in rats dosed with 0.3 mg/kg, levels of GH were significantly reduced and pulsatile GH secretion appeared ablated (p[lt]0.05, one way ANOVA with Dunnet[apos]s multiple comparison test). The dose of SXN101959 providing a 50% reduction in plasma GH levels ([sim]pIC[sub]50[/sub]) was 0.08 mg/kg in the first study and 0.04 mg/kg in the second study. These studies demonstrate that SXN101959 reproducibly produces a dose-dependent inhibition of pulsatile GH secretion in male rats after intravenous administration.[br][br]Disclosures: AM: Employee, SYNTAXIN. Nothing to Disclose: EAH, AW, SM, JB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1468 200 1652 SUN-669 PO18-01 Sunday 1449 2012


1446 ENDO12L_SUN-670 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Production of a Tamoxifen-Inducible, Cardiac-Specific Growth Hormone Receptor Disrupted Mouse Adam Jara, Xingbo Liu, Chance Benner, Edward O List, John J Kopchick Ohio University, Athens, OH; Heritage College of Osteopathic Medicine, Ohio University, Athens, OH; Ohio University, Athens, OH; College of Health Sciences and Professions, Ohio University, Athens, OH Intact growth hormone (GH) signaling is necessary for the proper development and maintenance of several tissues, including the heart. Evidence of the profound effect GH has on the myocardium can be found in patients with diseases that disrupt GH action. In pathological states of acromegaly (caused by oversecretion of GH), GH deficiency, and Laron Syndrome (a disease caused by mutations in the GH receptor) patients exhibit unique cardiac phenotypes of altered structure and function. The exact mechanisms responsible for these cardiac dysfunctions, however, are not well understood. To better understand the specific effects of GH action on cardiomyocytes, we have developed a tamoxifen-inducible, cardiac-specific GH receptor (GHR) disrupted mouse using a standard Cre-LoxP approach. Gene expression analysis using two-step reverse transcription quantitative polymerase chain reaction (RTqPCR) was performed to validate our ability to temporally, and spatially control GHR expression in a tamoxifen dependent manner. Sixteen week old male hemizygous Cre, homozygous GHR floxed ([alpha]MHC[sup]MerCreMer+/-[/sup]/GHR[sup]fl/fl[/sup]) mice and homozygous GHR floxed littermate controls (WT/GHR[sup]fl/fl[/sup]) were injected with either two intraperitoneal injections of tamoxifen in peanut oil (total dose 80mg/kg) or volume matched vehicle (n=3). All mice were dissected one week post induction and tissues were collected for RNA isolation. Initial results demonstrate that we are able to achieve approximately an 80% reduction in GHR mRNA expression specifically in cardiac tissue from [alpha]MHC[sup]MerCreMer+/-[/sup]/GHR[sup]fl/fl[/sup] mice treated with tamoxifen. Other tissues from [alpha]MHC[sup]MerCreMer+/-[/sup]/GHR[sup]fl/fl[/sup] and WT/GHR[sup]fl/fl[/sup] controls show no change in GHR expression levels when treated with tamoxifen or with vehicle. We believe this new mouse line will be a useful tool in elucidating the physiological and molecular impacts of impaired GH action in cardiomyocytes.[br][br]Sources of Research Support: This work was supported in part by the State of Ohio[apos]s Eminent Scholar Program that includes a gift from Milton and Lawrence Goll, by the AMVETS, by the Diabetes Research Initiative, by the Bill and Melinda Gates Foundation GMS program, by the Endocrine Society Summer Research Fellowship program and by NIH grants DK083729, AG031736.[br][br]Nothing to Disclose: AJ, XL, CB, EOL, JJK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1986 200 1653 SUN-670 PO18-01 Sunday 1450 2012


1447 ENDO12L_SUN-671 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) IGF-Independent Effects of Growth Hormone on Growth Plate Chondrocyte Function Shufang Wu, Francesco De Luca St Christopher[apos]s Hospital for Children, Philadelphia, PA [bold]Background[/bold]:[br]Evidence suggests that GH has IGF-1-independent effects on longitudinal bone growth. However, it is unclear whether these effects are mediated by IGF-2 in the growth plate.[br]Previously, we have shown that IGF-1 and GH facilitate growth plate chondrogenesis by activating NFkB-p65 in chondrocytes. Yet, we have not determined whether the effects of GH on NFkB-p65 activity are mediated by IGF-1.[br]Our goal was to evaluate the effects of GH on proliferation and differentiation, and on NFkB-p65 activity, in cultured growth plate chondrocytes transfected with IGF-1R si (short interfering) RNA.[br][bold]Results[/bold]:[br]After transfecting chondrocytes isolated from fetal (dpc 20) mouse metatarsal growth plates with IGF-1R siRNA, we demonstrated a significant reduction of IGF-1R mRNA (21.5 % of IGF-1R mRNA measured in control siRNA-transfected chondrocytes, assessed by real-time PCR) and protein (assessed by Western). We then cultured control siRNA- or IGF-1R siRNA-transfected chondrocytes with GH (10 ng/ml). In control siRNA-transfected chondrocytes, GH significantly induced thymidine incorporation (+ 144.8%, P[lt]0.01) and collagen X mRNA expression (3.4-fold, p[lt]0.01), compared to untreated control siRNA-transfected cells. Similarly, in IGF-1R siRNA-transfected chondrocytes, GH stimulated thymidine incorporation, (+ 132%, P[lt]0.01) and Collagen X mRNA expression (3.0-fold, p[lt]0.01), compared to untreated IGF-1R siRNA-transfected chondrocytes). Of note, there was no significant difference between control siRNA-transfected cells and IGF-1R siRNA-transfected cells with respect to the effects of GH on thymidine incorporation and collagen X mRNA expression.[br]We subsequently cultured control siRNA- or IGF-1R siRNA-transfected chondrocytes in the presence of 10 ng/ml GH, without or with PDTC (a known NF-kB inhibitor). GH induced NF-kB p65 activity in both control siRNA-transfected and IGF-1R siRNA-transfected (+214% and +207%, compared to untreated control siRNA- or IGF-1R siRNA-transfected chondrocytes, respectively. P[lt]0.01 for both comparisons). These effects were neutralized by the addition of PDTC in the culture medium (GH + PDTC vs. GH alone, in control siRNA-transfected cells: +117.3 % vs. +214 %, p[lt]0.01. In IGF-1R siRNA-transfected, +117.4 % vs. +207 %, p[lt]0.01).[br][bold]Conclusions[/bold]:[br]Our findings suggest that GH may modulate growth plate chondrocyte function via IGF-independent effects directly at the growth plate. Such effects may be mediated, at least in part, by NF-kB p65.[br][br]Nothing to Disclose: SW, FDL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 244 200 1654 SUN-671 PO18-01 Sunday 1451 2012


1448 ENDO12L_SUN-672 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Functional Interaction between Fibroblast Growth Factor 21 (FGF21) and Forkhead Box O 1 (Foxo1) in Growth Plate Chondrocytes Christopher Gibson, Shufang Wu, Francesco De Luca St Christopher[apos]s Hospital for Children, Philadelphia, PA [bold]Background:[/bold] We have previously demonstrated that the increased expression of Fibroblast Growth Factor 21 (FGF21) in the liver and in the growth plate during prolonged food restriction in mice causes Growth Hormone (GH) insensitivity. Previous evidence also indicates that the expression of the transcription factor Forkhead box O 1 (Foxo1) in the liver is induced during fasting, and it results in decreased GH action. Thus, we hypothesized that FGF21 may cause GH insensitivity in the growth plate by modulating the expression and activity of Foxo1.[br][bold]Results[/bold]: After 4 weeks of 50 % food restriction, in the tibial growth plate of wild-type (WT) mice the protein (by Western) and mRNA (by real-time PCR) expression of Foxo1 was greater than that in WT mice fed ad lib (mRNA expression: + 243 %, p[lt]0.05). In contrast, Foxo1 expression in food-restricted and fed ad-lib [italic]Fgf21[/italic] knock-out (KO) mice was similar. In the next experiment, we transfected chondrocytes isolated from fetal (dpc 20) metatarsal growth plates with control short interfering RNA (siRNA) or Foxo1 siRNA. [underline]In control siRNA-transfected cells[/underline], GH increased [sup]3[/sup]H-thymidine incorporation (+ 143 % of untreated cells, p[lt]0.01) and collagen X mRNA expression (+ 349 %, p[lt]0.01), with both effects being prevented by the addition of 100 ng/ml rhFGF21 in the culture medium (GH+rhFGF21 vs. GH alone: [sup]3[/sup]H-thymidine incorporation: +112 % vs. +143 % of untreated cells, p[lt]0.01; Collagen X expression: +192 % vs. +349 %, p[lt]0.05). rhFGF21 also decreased [[sup]125[/sup]I]-GH binding (GH+FGF21 vs. GH alone: + 83 % vs. + 127 % of untreated cells, p[lt]0.01).[br][underline]In Foxo1 siRNA-trasnfected cells[/underline], GH also stimulated [sup]3[/sup]H-thymidine incorporation (+150 % of untreated cells, p[lt]0.001) and collagen X mRNA expression (+268% of untreated cells, p[lt]0.01). The addition of rhFGF21 in the culture medium did not prevent the GH stimulatory effects (GH+FGF21 vs. GH alone: thymidine incorporation, + 151 % vs. + 150 %; Collagen X expression, + 329 % vs. + 270 %). In addition, rhFGF21 did not reduce [[sup]125[/sup]I]-GH binding (GH+FGF21 vs. GH alone: +142 % vs. + 137 %).[br][bold]Conclusions[/bold]: The increased expression of FGF21 in growth plate chondrocytes during prolonged food restriction inhibits the GH stimulatory effects on chondrocyte proliferation and differentiation by reducing GH receptor abundance on the chondrocyte cell membrane. Our findings indicate that the FGF21 effects on GH action are mediated, at least in part, by Foxo1.[br][br]Nothing to Disclose: CG, SW, FDL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 296 200 1655 SUN-672 PO18-01 Sunday 1452 2012


1449 ENDO12L_SUN-673 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) IGF-1 Receptor Modulates Acute Growth Hormone Sensitivity: Role of Protein Tyrosine Phosphatase Activity Yujun Gan, Yue Zhang, Jing Jiang, Stuart J Frank University of Alabama at Birmingham School of Medicine, Birmingham, AL; University of Alabama at Birmingham School of Medicine, Birmingham, AL; Birmingham VAMC, Birmingham, AL Growth hormone (GH) promotes growth and regulates metabolism. GH binds GH receptor (GHR), activating Janus kinase 2 (JAK2) and pathways including signal transducer and activator of transcription 5 (STAT5), thereby regulating expression of genes including insulin-like growth factor-1 (IGF-1). IGF-1 interacts with IGF-1 receptor (IGF-1R) to mediate some GH actions. This GH -[gt] GHR -[gt] IGF-1 -[gt] IGF-1R cascade represents the [ldquo]somatomedin hypothesis of GH action[rdquo]. Recently, we suggested alternative or complementary ways by which IGF-1R participates more proximally in GH signaling: 1) GH induces a GHR-JAK2-IGF-1R complex, independent of tyrosine phosphorylation; 2) IGF-1 cotreatment augments acute GH signaling; 3) IGF-1R deletion in osteoblasts or prostate cancer cells blunts acute GH signaling (1-3). In IGF-1R-deficient primary osteoblasts, adenovirally-driven IGF-1R re-expression normalizes GH-induced STAT5 phosphorylation, but re-expression of a truncated IGF-1R lacking its intracellular domain (ICD) (including its kinase domain) partially rescues GH-induced STAT5 activation and IGF-1 gene expression. Thus, IGF-1R ICD and kinase are dispensable for some IGF-1R-acute GH signaling collaboration. We now asked whether protein tyrosine phosphatase (PTP) activity influences GHR-IGF-1R collaboration. In osteoblasts, pre-incubation with orthovanadate, a general phosphatase inhibitor, rescued diminished GH-induced STAT5 activation resulting from IGF-1R deletion, suggesting involvement of phosphatase(s) in IGF-1R-acute GH signaling collaboration. Notably, a specific inhibitor of PTP-1B, which is known to be involved in GH signaling, also rescued GH signaling in IGF-1R-deleted cells, but inhibiting the SHP-1 and SHP-2 PTPs did not rescue the defect. The same inhibitor findings were observed in a separate IGF-1R knockdown system [ndash] stable expression of an IGF1R-directed shRNA in LNCaP, a human prostate cancer cell line. To independently test effects of PTP-1B inhibition, we adenovirally expressed a catalytically-inactive PTP1B (C215S mutation) in both osteoblasts and LNCaP. Similar to inhibitor results, dominant-negative PTP-1B expression reversed in both systems the defective GH-induced STAT5 activation in cells lacking IGF-1R. These data suggest that activity of PTP-1B or association of PTP-1B with a GH signaling pathway component(s) is enhanced upon IGF-1R deletion, an effect at least partially responsible for the resulting diminished GH-induced STAT5 activation.[br][br](1) Huang, Y., et al., Mol Endocrinol 2004; 18:1471. (2) Gan, Y., et al., Mol Endocrinol 2010; 24:644. (3) Zhang et al Abstract P3-233, 92nd Annual Endocrine Society Meeting, June, 2010.[br][br]Sources of Research Support: NIH R01DK46395.[br][br]Nothing to Disclose: YG, YZ, JJ, SJF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1244 200 1656 SUN-673 PO18-01 Sunday 1453 2012


1450 ENDO12L_SUN-674 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Ca2+ Deficiency-Induced and TRP Channel-Dependent IGF Signaling Stimulates Epithelial Cell Proliferation Wei Dai, Lisa Hebda, Xueping Zhong, Cunming Duan University of Michigan, Ann Arbor, MI Ca2+ deficiency causes abnormal epithelial cell proliferation and is a risk factor for colon cancer. The mechanisms by which Ca2+ deficiency leads to abnormal epithelial cell proliferation, however, are poorly understood. In this study, we elucidate a conserved signaling mechanism responsible for Ca2+ deficiency-induced epithelial cell proliferation using a unique animal model. The larval zebrafish skin contains a group of Ca2+ transporting epithelial cells or ionocytes. These cells are functionally equivalent to the mammalian intestinal Ca2+ transporting epithelial cells and contain all the molecular players for transcellular Ca2+ transport. We found that low [Ca2+] acclimation increases the skin ionocyte number by stimulating the proliferation of existing ionocytes. Low [Ca2+] resulted in a rapid and sustained activation of IGF signaling in these ionocytes exclusively. Inhibition of the IGF1 receptor, PI3K, and Akt, but not the MAPK, abolished the low [Ca2+]-induced epithelial proliferation. These cells specifically expressed the Ca2+ channel trpv5/6 gene and igfbp5a, a gene encoding a high-affinity binding protein for IGFs. Knockdown of igfbp5a attenuated IGF signaling. Blocking Trpv5/6 abolished the low [Ca2+]-induced IGF signaling and ionocyte proliferation. Surprisingly, the Trpv5/6-dependent activation of IGF signaling is independent from intracellular Ca2+/CaM-dependent signaling. Rather, it is mediated through Trpv5/6-mediated membrane depolarization. A similar mechanism was observed in human colon epithelial cells. These results unravel an evolutionarily conserved and Ca2+ deficiency-induced signaling module linking a TRP channel to growth factor signaling and a co-opted role of IGF signaling in epithelial proliferation under calcium deficient conditions.[br][br]Sources of Research Support: This work was supported in part by NSF Grant IOB-0543018 and IOS-1051034.[br][br]Nothing to Disclose: WD, LH, XZ, CD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2338 200 1657 SUN-674 PO18-01 Sunday 1454 2012


1451 ENDO12L_SUN-675 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Differential Gene Expression Studies in Insulin-Like Growth Factor II Knockout and Wild-Type Embryos Mary Frances Lopez, Reddy Gali, Sandford B Church, Petra Bukovac-Kempna, Joel N Hirschhorn Boston Children[apos]s Hospital, Boston, MA; Harvard Medical School, Boston, MA; Harvard Medical School, Boston, MA; Harvard Medical School, Boston, MA Insulin-like growth factor II (IGF-II) is a hormone that plays an important role in fetal life. Overexpression of the IGF-II gene has been described in Wilms tumor and Beckwith-Wiedemann syndrome. Targeted disruption of the mouse IGF-II gene (IGF-II KO) results in reduction in fetal growth. IGF-II KO mice are born 60% the size of their normal littermates. This decrease in overall size is thought to be due to a reduction in the rate of mitosis during embryogenesis. The goal of this study was to examine the mechanisms by which IGF-II promote fetal growth in the mouse embryo by doing differential gene expression studies between the IGF-II knockout (IGF-IIKO) and wild-type (WT) littermate fetuses. Embryos were collected on day 14 of gestation. RNA was extracted using the RNeasy Mini Kit for processing and hybridization using the Illumina Mouse-WG6 BeadChips (Boston Children[apos]s Hospital Molecular Genetics Microarray Core Facility). Microarray analysis revealed many differentially expressed genes in the IGF-II KO embryo. Among those genes we have confirmed by Q-PCR an 8 fold up-regulation of Cathepsin E and the HAMP genes (P[lt] 0.0001), which have been linked to fetal development and neoplasia. Results from these experiments will provide an insight into the biology of fetal growth and potentially to IGF-II related neoplasia.[br][br]Sources of Research Support: This study was supported by an NIDDK grant (5R01DK075787).[br][br]Nothing to Disclose: MFL, RG, SBC, PB-K, JNH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2084 200 1658 SUN-675 PO18-01 Sunday 1455 2012


1452 ENDO12L_SUN-676 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Influence of GH Receptor and PRL Receptor on GH and PRL Signaling in Human Breast Cancer Cells Dongmei Sun, Jie Xu, Jing Jiang, Serge Y Fuchs, Stuart J Frank University of Alabama at Birmingham School of Medicine, Birmingham, AL; University of Pennslyvania School of Veterinary Medicine, Philadelphia, PA; University of Alabama at Birmingham School of Medicine, Birmingham, AL; Birmingham VAMC, Birmingham, AL Growth hormone (GH) and prolactin (PRL) are closely related pituitary peptide hormones. GH receptor (GHR) and PRL receptor (PRLR) share structural similarities and common signaling mechanisms; each couples to the non-receptor tyrosine kinase, JAK2, and activates pathways including STAT5 and ERKs. In humans, GH promotes growth and enhances muscle mass, in addition to regulating carbohydrate and lipid metabolism. PRL is most known for impacting mammary gland development and lactogenesis. In addition to these classical functions, recent findings in rodents suggest both GH and PRL may impact breast cancer initiation, progression, or invasion. One interesting facet of GH/PRL biology is that human GH interacts with both human GHR and PRLR, while human PRL binds human PRLR, but not human GHR. Herein, we examined the roles of GHR and PRLR in transducing GH and PRL signals in the human breast cancer cell line, T47D. We prepared T47D cells stably expressing shRNA against GHR (T47D-GHR-KD), PRLR (T47D-PRLR-KD), or encoding a scrambled control sequence (T47D-SC). Control cells, like parental T47D cells, express immunologically-detectable GHR and PRLR and respond acutely (15 min) dose-dependently to GH or PRL with STAT5 and ERK phosphorylation. T47D-PRLR-KD cells and T47D-GHR-KD cells display markedly reduced PRLR and GHR, respectively, compared to T47D-SC cells, as assessed by immunoblotting. As expected, T47D-PRLR-KD cells exhibit markedly reduced PRL-induced STAT5 and ERK phosphorylation compared to control cells; however, GH-induced signaling was greatly augmented with a left-shifted dose-response curve. Remarkably, T47D-GHR-KD cells, which nearly completely lack GHR protein, retained their GH sensitivity in terms of acute STAT5 and ERK activation; moreover, PRL-induced signaling was also increased in these cells. Analysis to date of potential mechanisms indicates that GHR protein (but not mRNA) abundance is augmented in T47D-PRLR-KD cells and this is related to an enhanced half-life of the mature cell-surface GHR in these cells. Similarly, PRLR protein abundance is augmented in T47D-GHR-KD cells. These novel findings indicate reduction in either GHR or PRLR in human breast cancer cells enhances acute GH sensitivity and suggest these two receptors reciprocally interact to modulate GH sensitivity. Further mechanistic and functional studies are underway to better understand pathophysiological and therapeutic implications of these findings.[br][br]Sources of Research Support: NIH R01 DK58259.[br][br]Nothing to Disclose: DS, JX, JJ, SYF, SJF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1338 200 1659 SUN-676 PO18-01 Sunday 1456 2012


1453 ENDO12L_SUN-677 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Somatostatin and Cortistatin Play Differential Roles in the Regulation of Growth Hormone (GH)/IGF-I System Expression in the Mammary Gland of Obese Mice Raul Miguel Luque, Alicia Villa-Osaba, Manuel David Gahete, Jose Cordoba-Chacon, Ana Isabel Pozo-Salas, Laura Maria Lopez-Sanchez, Marina Alvarez-Benito, Francisco Gracia-Navarro, Luis de Lecea, Jose Lopez-Miranda, Rhonda Denise Kineman, Justo Pastor Castano University of C[oacute]rdoba, C[oacute]rdoba, Spain; University of Illinois at Chicago, Chicago, IL; Hospital Univ Reina Sof[iacute]a, C[oacute]rdoba, Spain; Stanford University, Stanford, CA; Hospital Univ Reina Sof[iacute]a, Instituto Maim[oacute]nides de Investigaci[oacute]n Biom[eacute]dica de C[oacute]rdoba (IMIBIC), CIBERObn, C[oacute]rdoba, Spain Locally produced GH and IGF-I are key factors in the regulation of mammary gland (MG) development and may be important in breast cancer development and progression. Somatostatin (SST) and cortistatin (CORT) act in concert to regulate GH/IGF-I axis at various levels but their role in regulating these systems in MG is unknown. Since obesity markedly alter the expression pattern of these systems in different tissues and is closely associated to MG (patho)physiology, we sought to investigate the role of SST and CORT in regulating GH/IGF-I system expression in MG and its relationship with obese status. We have analyzed the gene expression profiles of GH/IGF-I system in the MG of SST- or CORT-knockout (KO) mice and their respective littermate controls fed either a low fat (LFD) or a high fat (HFD) diet for 16wks. Our results show that, under LFD, lack of SST or CORT does not influence the expression pattern of GH/IGF-I system expression in the MG. However, in the MG of obese mice, where GH/IGFI system [GH receptor (GHR) and IGF-I and its receptor (IGFIR)] is upregulated, lack of SST partially suppressed but lack of CORT completely blunted this upregulation. SST/CORT system is known to be a negative regulator of GHR, IGF-I and IGFIR expression in different tissues and, consequently, a further elevation of GH/IGF-I axis in the MG of obese SST- and CORT-KO mice was expected. Interestingly, expression of SST/CORT receptor sst2 was significantly upregulated in the MG of obese CORT-KO mice while a non-significantly elevation was observed in SST-KO mice. These results suggest that the compensatory elevation in sst2 expression observed in the MG of obese CORT-KO mice could be sensitizing the MG to endogenous SST and, in turn, blocking obesity-induced upregulation of GH/IGF-I system. However, in obese SST-KO mice where sst2 is not as prominently elevated, endogenous CORT cannot fully counteract obesity-induced increase in GH/IGF-I system. Alternatively, it could be argued that SST is more potent than CORT in suppressing GH/IGF-I system in the MG. These results offer new information on the factors involved in the dysregulation of endocrine/metabolic homeostasis of MG in obesity, and also suggest that CORT is not a simple analog of SST in regulating MG physiology. Altogether, our findings may provide new cues to identify novel molecular targets for diagnosis and/or treatment of MG pathologies including breast cancer.[br][br]Sources of Research Support: This work was supported by the following grants: BFU2010-19300, RYC-2007-00186, CIBERObn (MICINN/FEDER), BIO-139, CTS-5051 (Junta de Andaluc[iacute]a) (R.L.M; F.G.N; J.P.C); Fundaci[oacute]n Caja Madrid (M.D.G.); Fundaci[oacute]n Alfonso Martin Escudero (J.C.C); JCI-2009-05675 (L.L-S); Department of Veterans Affairs, NIH R01DK088133 (R.D.K.); NIH grant (to L. dD).[br][br]Nothing to Disclose: RML, AV-O, MDG, JC-C, AIP-S, LML-S, MA-B, FG-N, LdL, JL-M, RDK, JPC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1970 200 1660 SUN-677 PO18-01 Sunday 1457 2012


1454 ENDO12L_SUN-678 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Evidence That the Strong Physiological Downregulation of[italic] Igf2 [/italic]in Postnatal Tissues Is Driven by Downregulation of [italic]E2f1[/italic] and [italic]3[/italic] Julian C Lui, Jeffrey Baron National Institute of Child Health and Human Development, Bethesda, MD Mice with targeted Igf2 disruption exhibit severe growth retardation at birth, indicating that this growth factor is critical for normal somatic growth. Igf2 mRNA is highly expressed in multiple tissues prenatally and in early postnatal life but then is dramatically downregulated by 4 wks of age. These observations suggest that Igf2 supports the rapid proliferation of developing organs in early life, and its subsequent downregulation contributes to the progressive slowing of cell proliferation and body growth during later juvenile life. The mechanism responsible for the robust downregulation of Igf2 expression in postnatal organs is unknown. Sequence analysis revealed that two E2F consensus binding motifs were present in the promoter regions of IGF2 in both human and mouse. We therefore hypothesized that the downregulation of Igf2 is caused by declining effects of the E2f family of transcription factors.[br]Expression microarray in mice showed that, of the 8 [italic]E2f [/italic]family members, E2f1 and [italic]E2f3[/italic] were significantly downregulated from 1 to 4 wks of age in kidney, lung, and heart (all [italic]P [/italic][lt] 0.001, [gt]1.5-fold). These findings were confirmed by real-time PCR (all [italic]P [/italic][lt] 0.05) and western blot in multiple organs at 1, 4, and 8 wks. Using chromatin immunoprecipitation, we next showed that in mouse kidney and lung, E2f3 binds to the E2F consensus motifs in the [italic]Igf2[/italic] promoter at 1 wk of age ([italic]P[/italic] [lt] 0.001), but this E2f3 binding decreases by 4 wks of age ([italic]P[/italic] [lt] 0.001). The decreased [italic]E2f3 [/italic]binding with age may thus drive the downregulation of [italic]Igf2[/italic]. Next, to confirm that [italic]E2f[/italic] promotes[italic] Igf2[/italic] expression, [italic]E2f1[/italic] or 3 was overexpressed in primary hepatocytes isolated from fetal (E16) and adult (5-wk-old) mice and Igf2 expression was studied. Fetal hepatocytes expressed high levels of [italic]Igf2 [/italic]and showed minimal response to the overexpression of [italic]E2f1[/italic] or [italic]3[/italic]. In contrast, adult hepatocytes expressed low levels of [italic]Igf2[/italic], and overexpression of [italic]E2f1[/italic] or [italic]3[/italic] caused [gt]25-fold increase in [italic]Igf2[/italic] expression (both P[lt]0.001).[br]In summary, during early postnatal life, we observed a decline in [italic]E2f1[/italic] and [italic]3[/italic] expression and a decline in binding of [italic]E2f3[/italic] to the [italic]Igf2 [/italic]promoter. In cultured adult hepatocytes, overexpression of [italic]E2f1[/italic] or [italic]3[/italic] partially restored the expression of [italic]Igf2[/italic]. Taken together, our findings suggest that the robust downregulation of[italic] Igf2 [/italic]in juvenile organs is driven by the postnatal downregulation [italic]E2f1[/italic] and [italic]3[/italic].[br][br]Nothing to Disclose: JCL, JB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1639 200 1661 SUN-678 PO18-01 Sunday 1458 2012


1455 ENDO12L_SUN-679 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Growth Hormone (GH)-Induced STAT5 Signaling in Liver of Growing Mice Ana Isabel Sotelo, Carolina Soledad Martinez, Johanna Gabriela Miquet, Veronica Gabriela Piazza, Laura Daniela Ratner, Susana Beatriz Rulli, Lorena Gonzalez, Daniel Turyn IQUIFIB- Instituto de Qu[iacute]mica y Fisicoqu[iacute]mica Biol[oacute]gicas, Buenos Aires, Argentina; Ibyme-Instituto de Biolog[iacute]a y Medicina Experimental, Buenos Aires, Argentina Growth hormone (GH) is the principal endocrine regulator of postnatal body growth. Signal transducer and activator of transcription 5b (STAT5b) is the main GH-signaling mediator, related to IGF1 synthesis and somatic growth. Glucocorticoid receptor (GR) and hepatocyte nuclear factor 1 (HNF1) are STAT5 coactivators, whereas GH-induced suppressors of cytokine signaling (SOCS-2,-3 and CIS) and phospho-tyrosine phosphatases (PTP-1B, SHP1 and SHP2) contribute to signaling termination. Rodents exhibit two instances of rapid postnatal growth: the first is perinatal and independent of GH, the second takes place around weaning and is controlled by GH. The objective of the present study was to assess differential sensitivity to growth hormone during the growth period. For that purpose, GH-induced STAT5 signaling and the ontogeny of modulators that regulate this pathway were determined in the liver of growing mice. Three representative ages were chosen: 1 week animals [ndash]in the GH-independent phase of growth-, 2.5 week mice [ndash]at the onset of the GH-dependent phase of growth-, and 9 week young adults, used as reference. Since GH secretion is sexually dimorphic, both genders were analyzed in parallel. Animals received an ip GH bolus (1[micro]g/g BW) or saline and were sacrificied 7.5 min later. Liver protein content was assessed by immunoblotting and by immunohistochemistry. One-week-old animals presented low GH-induced STAT5 phosphorylation, associated with lower hepatic receptor content and with high levels of the signaling suppressor CIS and phosphatase PTP-1B, whereas the abundance of glucocorticoid receptor and HNF-1 was low. Maximal GH-induced STAT5 activation was found for 2.5 week animals, which could be related with higher STAT5 protein levels at that age, and with an age-dependent decline of CIS and PTP-1B. On the other hand, SOCS2 and SOCS3 levels gradually increased to achieve higher levels in the 9 week control than at earlier ages. GR and HNF1 also exhibited a maximum at adulthood, while phosphatases SHP1 and 2 did not present any age variation. Although GH secretion is sexually dimorphic, no gender difference was observed for the proteins studied. We conclude that GH-induced STAT5 signaling presents age-dependent activity in the liver of growing mice, with its maximum coinciding with the onset of the GH-dependent phase of growth, accompanied by age-related changes in the pathway modulators.[br][br]Sources of Research Support: ANPCyT(Agencia Nacional de Promoci[oacute]n Cient[iacute]fica y Tecnol[oacute]gica), UBACyT (Universidad de Buenos Aires) and CONICET (Consejo Nacional de Investigaciones Cient[iacute]ficas y Tecnol[oacute]gicas).[br][br]Nothing to Disclose: AIS, CSM, JGM, VGP, LDR, SBR, LG, DT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 799 200 1662 SUN-679 PO18-01 Sunday 1459 2012


1456 ENDO12L_SUN-680 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Low-Carb/High-Fat Diets Lead to Lower IGF-I Levels and Reduced Longitudinal Growth in Rats Regardless if the Dietary Fat Source Derives from Beef Tallow or Soybean Oil Chrysanthi Taxeidi, Maximilian Bielohuby, Sarina Meurer, Amon Horngacher, Martin Bidlingmaier Medizinische Klinik und Poliklinik IV, Klinikum der LMU, Munich, Germany Ketogenic low-carbohydrate/high-fat diets (LC-HFD) are used as part of treatment for epilepsy in children, but have been shown to negatively affect longitudinal growth. In a rat model we have previously shown that a ketogenic LC-HFD led to increased fat accumulation and reduced lean body mass (LBM), paralleled by low IGF-I levels. Furthermore, LC-HFD was associated with impaired longitudinal growth and reduced osteoblast activity. As in this study the only dietary fat source was beef tallow, we now wanted to investigate whether the use of soybean oil [ndash]frequently claimed to be the [ldquo]healthier[rdquo] fat [ndash] would ameliorate the negative effects of ketogenic LC-HFD. We therefore pair-fed male 12-wk-old Wistar rats (n=7/group) iso-energetic amounts of either beef tallow based LC-HFD (LC-HFD-B; high in saturated and mono unsaturated fatty acids (FA)) or soybean oil based LC-HFD (LC-HFD-S; high in poly unsaturated FA) or standard chow diet, also based on either beef tallow (CH-B) or soybean oil (CH-S) (diet composition in % of metabolizable energy, fat/protein/CHO: LC-HFD-B and LC-HFD-S (78.7/19.1/2.2), CH-B and CH-S (16.7/19/64.3). After 4 weeks on the respective diets, rats were sacrificed. We measured bodyweight (BW), nose-to-rump, tibia- and femur lengths and LBM. Fat pads were extracted and weighed and serum IGF-I was measured by immunoassay (IDS, Boldon, UK). As expected, LC-HFD-B led to significantly lower terminal BW when compared to CH-B. The same was observed when using soybean oil instead of beef tallow (CH-S: 385.9[plusmn]6.9 LC-HFD-S: 344[plusmn]8.8g, p[lt]0.05). Furthermore, LC-HFD-S fed rats were shorter (CH-S: 21.43[plusmn]0.3cm, LC-HFD-S: 20.4[plusmn]0.2cm, p[lt]0.05), and had shorter tibiae and femur bones. Epididymal fat pad weights were higher with LC-HFD-S (p=0.05) when compared to CH-S but not when compared with LC-HFD-B (CH-S: 3.8[plusmn]0.4g, LC-HFD-S: 4.8[plusmn]0.5g; CH-B: 3.4[plusmn]0.4g; LC-HFD-B: 5.3[plusmn]0.3g). LBM was lower with LC-HFD-S (CH-S: 179.8[plusmn]4.3g, LC-HFD-S: 159.6[plusmn]2.6g; p[lt]0.01), but not significantly different between LC-HFD-S and LC-HFD-B. IGF-I levels were lower by about 42% also with LC-HFD-S when compared to CH-S (p[lt]0.001).[br]In summary, usage of soybean oil based LC-HFD which is rich in poly unsaturated FA instead of using beef tallow as fat source did not prevent any of the negative effects on body composition and longitudinal growth. Furthermore, LC-HFD dramatically inhibit the GH/IGF system independent of the fat source.[br][br]Disclosures: MB: Study Investigator, Chiasma. Nothing to Disclose: CT, MB, SM, AH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 961 200 1663 SUN-680 PO18-01 Sunday 1460 2012


1457 ENDO12L_SUN-681 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Growth Hormone Inhibits Differentiation of Bovine Preadipocytes into Adipocytes Partly through Signal Transducer and Activator of Transcription 5 Lidan Zhao, Aihua Wang, Honglin Jiang Virginia Tech, Blacksburg, VA Growth hormone (GH) is known to inhibit adipogenesis in animals, but little is known about the underlying mechanism. In this work, we determined the effect of GH on the differentiation of primary bovine preadipocytes into adipocytes and if this effect is mediated by signal transducer and activator of transcription 5 (STAT5), a transcription factor known to be activated by signaling from the GH receptor. We derived preadipocytes from bovine adipose tissue explants and induced them to differentiate into adipocytes in the presence of GH (100 ng/ml), adenovirus expressing constitutively active STAT5b (STAT5bCA, 100 multiplicity of infection), or a combination of GH and the synthetic STAT5 inhibitor N[prime]-((4-Oxo-4H-chromen-3-yl)methylene)nicotinohydrazide (100 [mu]M). Differentiation of preadipocytes into adipocytes was assessed by Oil Red O staining and by measuring glycerol-3-phosphate dehydrogenase (G3PDH) activity, rate of acetate incorporation, and mRNA expression of adipogenic markers CCAAT/enhancer binding protein [alpha] (CEBP[alpha]) and peroxisome proliferator-activated receptor [gamma](PPAR[gamma]). Oil Red O staining showed that fewer preadipocytes became adipocytes in the presence of GH than in the absence of GH. The preadipocytes treated with GH had lower G3PDH activity, lower acetate incorporation rate, and lower CEBP[alpha] and PPAR[gamma] mRNA expression than those without GH treatment. Similarly, the preadipocytes cultured in the presence of STAT5bCA adenovirus had lower G3PDH activity, lower acetate incorporation rate, and lower expression of CEBP[alpha] mRNA than those cultured in the presence of control adenovirus. Addition of the STAT5 inhibitor blocked GH-induced phosphorylation of STAT5 and GH-induced decreases in expression of CEBP[alpha] and PPAR[gamma] mRNAs. Taken together, these results demonstrate that GH inhibits differentiation of primary bovine preadipocytes into adipocytes in culture and suggest that this effect of GH is mediated at least in part through STAT5.[br][br]Nothing to Disclose: LZ, AW, HJ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1643 200 1664 SUN-681 PO18-01 Sunday 1461 2012


1458 ENDO12L_SUN-682 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) GH and Not IGF-1 Decreases Hepatic Production of Thyroxine-Binding Globulin (TBG) and Reduces Circulating Thyroxine (T4) in Diet-Induced Obese Mice Edward O List, Darlene E Berryman, John D Blischak, Bryan Tysl, Laura Kutz, Adam Jara, W Edward Visser, Theo J Visser, John J Kopchick Ohio University, Athens, OH; Ohio University, Athens, OH; Ohio University, Athens, OH; Erasmus University Medical Center, Rotterdam, Netherlands; Ohio University, Athens, OH Previously our laboratory investigated the effects of GH alone, IGF-1 alone or GH and IGF-1 in combination in obese mice fed a high fat diet. Three weeks of GH treatment improved body composition (increasing lean mass and decreased fat mass) and normalized hyperglycemia and glucose intolerance. Three weeks of IGF-1 treatment increased lean mass but failed to improve hyperglycemia and glucose intolerance. In the current study, these mice were utilized to study hepatic gene expression, and serum thyroid hormone levels were also measured.[br]Thirty-nine C57BL/6J mice were placed on a high fat (HF) diet for 16 weeks to induce obesity and diabetes. Starting at week 16, the HF fed mice were divided into 4 groups and treated twice daily for 3 weeks with saline, 5ug rbGH, 2.5ug rhIGF-1, or both 5ug rbGH and 2.5ug rhIGF-1. One group remained on standard chow and served as LF controls, which were also injected twice daily with saline. Hepatic gene expression was determined from livers of mice treated with either GH or IGF-1 using Agilent 4 x 44K Whole Mouse Genome MicroArrays. Real-time RT-PCR was performed on hepatic RNA to verify changes in gene expression. Fasting serum samples were shipped to Erasmus University Medical Center, Rotterdam, NETHERLANDS to determine total circulating thyroxine (T4) and triiodothyronine (T3) levels.[br]A variety of hepatic genes were determined to be altered with GH alone and IGF-1 alone treatments. Since few IGF-1 receptors are present in liver, the majority of GH[apos]s effect on liver should be independent of IGF-1. This was evident in that only [sim]5% of the genes found to be significantly altered with GH and IGF-1 treatments were regulated in a similar manner and none of the top 10 decreased or increased genes were shared between GH and IGF-1 treatments. For GH treatment, the single most down regulated hepatic gene was found to be thyroxine-binding globulin (TBG). Real-time RT-PCR verified that GH significantly decreased TBG in liver. GH treatment also resulted in a significant reduction in circulating T4 and slight but non-significant decrease in T3. IGF-1 treatment did not alter levels of T3 or T4. In conclusion, we have demonstrated that GH significantly reduces TBG hepatic gene expression in obese mice. The precise role of reduced expression of TBG on physiology during GH treatment remains unclear.[br][br]Sources of Research Support: This work was supported in part by the State of Ohio[apos]s Eminent Scholar Program that includes a gift from Milton and Lawrence Goll, by the AMVETS, by the Diabetes Research Initiative at Ohio University, and by NIH grants DK083729, AG031736.[br][br]Nothing to Disclose: EOL, DEB, JDB, BT, LK, AJ, WEV, TJV, JJK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1954 200 1665 SUN-682 PO18-01 Sunday 1462 2012


1459 ENDO12L_SUN-683 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Insulin-Induced Growth Hormone Receptor Expression in Grass Carp Hepatocytes: Functional Role of MAPK and PI3K/Akt Cascades Quan Jiang, Gerald F Brown, Mulan He, Anderson OL Wong The University of Hong Kong, Hong Kong, China In mammals, insulin can potentiate growth hormone (GH)-induced IGF-I secretion and gene expression at the hepatic level but the mechanisms involved have not been characterized. Recently, the synergy of insulin and GH on IGF-I gene expression has been demonstrated in grass carp, implying that the phenomenon is well conserved during vertebrate evolution. To shed light on the role of GH receptor (GHR) in insulin potentiation of IGF-I gene expression induced by GH in the carp liver, the effects of insulin on GHR expression were examined in primary culture of grass carp hepatocytes. In this case, GHR mRNA and protein levels were both elevated by insulin treatment with rapid phosphorylation of insulin receptor, MEK1/2, ErK1/2, P38 MAPK and Akt, respectively. The stimulatory effect on GHR mRNA expression could be negated by inactivating insulin receptor but not IGF-I receptor. Insulin-induced GHR mRNA and protein expression were also sensitive to pharmacological inhibition of MEK1/2, P38 MAPK, PI3K, Akt and mTOR. In carp hepatocytes, insulin did not alter the half-life of GHR mRNA, suggesting that the effect of insulin was exerted at the transcriptional level rather than post-transcriptional modification of transcript stability. This idea is consistent with the results based on aT3 cells transfected with a LUC reporter carrying the 5[apos]promoter of carp GHR, in which insulin treatment could up-regulate GHR promoter activity via transactivation of cis-acting elements located in the proximal promoter downstream of position -331. Similar to GHR mRNA expression in carp hepatocytes, insulin-induced GHR promoter activity could be blocked by inhibiting MEK1/2, P38 MAPK, PI3K, Akt and mTOR, respectively. These results, as a whole, suggest that insulin can induce GHR expression in the carp liver via activation of GHR gene transcription through the MAPK- and PI3K/Akt-dependent pathways, which may contribute to the potentiating effect of insulin on GH-induced IGF-I gene expression.[br][br]Sources of Research Support: GRF grants from Research Grant Council (Hong Kong).[br][br]Nothing to Disclose: QJ, GFB, MH, AOLW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 125 200 1666 SUN-683 PO18-01 Sunday 1463 2012


1460 ENDO12L_SUN-684 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Inflammation of the Sympathetic Nervous System (SNS) in Type 1 Diabetes: Implications for a Role for IGF-1 and IGFBP-3 Maria B Grant, Ping Hu, Jeffery S Thinschmidt, Sugata Hazra, Yuanqing Yan, Sergio Caballero, Michael E Boulton, Daniel R Saban, Tailoi Chan-Ling University of Florida, Gainesville, FL; University of Sydney, Sydney, Australia; Harvard Medical School, Boston, MA; University of Florida, Gainesville, FL Increased IGFBP-3 expression reduced the number of activated microglial cells and apoptosis of neuronal cells in the retina following oxygen induced injury (Kielczewski JL 2011). We asked whether IGFBP-3 is playing a role in type 1 diabetes (T1D)-associated inflammation in the CNS and retina. In brain and retina of T1D mice, we investigated microglia activation and monocyte subpopulations and IGF-1 and IGFBP-3 immunoreactivity was assessed in neurons. Mice underwent bone marrow (BM) transplantation with green fluorescent protein (GFP) cells followed by induction of diabetes with streptozotocin to trace BM-derived cell infiltration into brain regions and retina. In mice with 4-42 weeks of T1D, activated microglia, identified by staining for the actin-binding protein Iba-1, ED1, [italic]GS[/italic] isolectin B4 and by morphology, were found in increased numbers in the retina, hypothalamus (HYPO) and granular insular cortex (GIC) but not the motor cortex (MC) and basal nucleus (BN). The density of NeuN[sup]+[/sup] neurons in HYPO in control mice was approximately 1255/mm[sup]2[/sup] and did not change in diabetic mice. The mean size of NeuN[sup]+[/sup] cell was 86 [micro]m[sup]2[/sup] in control mice, and reduced to 70 [micro]m[sup]2 [/sup]in diabetic mice (p=0.03). In both control and diabetic brain, almost all IGF-I[sup]+[/sup] cells were NeuN[sup]+[/sup] (indicative of neurons) and were found in the HYPO region. The HYPO is the central regulator of SNS. In this region, the density of IGF-I[sup]+ [/sup]cells was reduced from 500/mm[sup]2[/sup] in control to 440/mm[sup]2[/sup] (P=0.037) in diabetic mice. In contrast, IGFBP-3[sup]+[/sup]/NeuN[sup]+[/sup] cells were distributed across the brain and their density was increased in the diabetic compared to control brain. The density of IGFBP-3[sup]+[/sup] cells in HYPO in control mice was approximately 820/mm[sup]2[/sup] and increased to 1227/mm[sup]2[/sup] in diabetic mice; all NeuN[sup]+[/sup] neurons became IGFBP-3[sup]+[/sup] in diabetic mice. Treatment of T1D mice with the anti-inflammatory agent minocycline, which crosses the blood brain barrier and blood retinal barrier, reduced activated Iba-1[sup]+[/sup] microglia by about 20% and infiltration of CD45[sup]+[/sup]/CCR2[sup]+[/sup]/GR-1[sup]+[/sup] bone-marrow-derived monocytes by 86% in these SNS centers, and similarly reduced retinal monocytes by 90%. We conclude that in T1D, inflammation of SNS occurs as it does in traditional target tissues of diabetic complications such as the retina. Minocycline may represent novel strategy for management of T1D induced inflammation. Increases in IGFBP-3 expression by neurons may represent an endogenous mechanism to suppress diabetes-induced inflammation in diabetes.[br][br](1) Kielczewski JL, Li Calzi S, Shaw LC, Cai J, Qi X, Ruan Q, Wu L, Liu L, Hu P, Chan-Ling T, Mames RN, Firth S, Baxter RC, Turowski P, Busik JV, Boulton ME, Grant MB. Free insulin-like growth factor binding protein-3 (IGFBP-3) reduces retinal vascular permeability in association with a reduction of acid sphingomyelinase (ASMase). Invest Ophthalmol Vis Sci. 2011 Oct 21;52(11):8278-86. Print 2011. PubMed PMID: 21931131; PubMed Central PMCID: PMC3208060.[br][br]Sources of Research Support: NEI RO1 EY012601, NEI RO1 007739, American Heart Association.[br][br]Nothing to Disclose: MBG, PH, JST, SH, YY, SC, MEB, DRS, TC-L 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2103 200 1667 SUN-684 PO18-01 Sunday 1464 2012


1461 ENDO12L_SUN-685 POSTER SESSION: GH [amp] Growth Factors: Normal [amp] Abnormal Regulation [amp] Actions (1:30 PM-3:30 PM) Balance of TACE and TIMP3 Determines the Susceptibility of Cells to Phorbol Ester-Induced GH Receptor Proteolysis and Desensitization to GH Yue Zhang, Kimberly Loesch, Jing Jiang, Stuart J Frank University of Alabama at Birmingham School of Medicine, Birmingham, AL; University of Alabama at Birmingham School of Medicine, Birmingham, AL; Birmingham VAMC, Birmingham, AL Growth hormone (GH) binds the cell surface GH receptor (GHR) extracellular domain (ECD) and elicits important anabolic and metabolic actions. We demonstrated that metalloproteolytic cleavage in the GHR perimembraneous ECD stem releases the ECD, leaving behind a transmembrane (TMD)- and intracellular domain (ICD)-containing remnant. This cleavage, catalyzed by TNF-a converting enzyme (TACE/ADAM17), is induced in vitro by serum, platelet-derived growth factor, or the phorbol ester, PMA and renders cells less sensitive subsequently to GH (1,2). [italic]In vivo[/italic] work in mice suggests that endotoxin also elicits proteolytic hepatic GHR downregulation and desensitization to GH (3). Tissue inhibitor of metalloproteinases 3 (TIMP3) is an endogenous specific TACE inhibitor. C14 cells are human fibrosarcoma cells stably expressing GHR and JAK2. PMA treatment of C14 induces GHR proteolysis characterized by loss of GHR and accumulation remnant detected by immunoblotting. HEK-293-GHR-JAK2 cells are human epithelial cells stably expressing GHR and JAK2. As in C14, GH treatment activated GHR, JAK2, and STAT5 in HEK-293-GHR-JAK2. In contrast to C14, minimal PMA-inducible GHR proteolysis was observed in HEK-293-GHR-JAK2. Endogenous TIMP3 was easily detected by immunoblotting in HEK-293-GHR-JAK2, but not in C14. We therefore tested effects of TIMP3 expression on GHR proteolysis in C14, achieved by infection with an adenovirus encoding TIMP3. Ad-TIMP3-mediated increased TIMP3 expression was confirmed by immunoblotting and PMA-induced GHR proteolysis was dramatically reduced. Notably, in control adenovirus-infected cells, GH-induced STAT5 phosphorylation was decreased by [sim]80% when the cells were exposed to PMA compared to vehicle alone, consistent with our previous findings. However, in Ad-TIMP3-infected cells, GH-induced STAT5 phosphorylation was largely preserved ([sim]90%), despite PMA exposure. We are currently testing effects of siRNA-mediated TIMP3 knockdown in HEK-293-GHR-JAK2 cells on PMA-induced GHR proteolysis and GH sensitivity. There are two forms of TACE [ndash] full-length precursor and mature TACE lacking the prodomain, removal of which is believed required for TACE activation. By immunoblotting, the ratio of mature/precursor TACE was greater in C14 than HEK-293-GHR-JAK2 (1.23 vs 0.56). These data suggest the relative balance of TACE, especially mature TACE, and TIMP3 affects susceptibility of GHR to PMA-induced proteolysis and the degree of PMA-induced desensitization to GH.[br][br](1) Zhang Y et al., Endocrinology. 2000; 141: 4342. (2) Guan R et al., Endocrinology. 2001; 142: 1137. (3) Wang X et al., Mol Endocrinol. 2008; 22: 1427.[br][br]Sources of Research Support: VA Merit Review Award (SJF).[br][br]Nothing to Disclose: YZ, KL, JJ, SJF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1235 200 1668 SUN-685 PO18-01 Sunday 1465 2012


1462 ENDO12L_SUN-686 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) GABA Inhibition of Kisspeptin Release in Prepubertal Monkeys: Implications on the Mechanism of the Pubertal Increase of GnRH Release Joseph R Kurian, Kim L Keen, Kathryn A Guerriero, Ei Terasawa University of Wisconsin, Madison, WI; University of Wisconsin, Madison, WI Previously, we have shown that a reduction in GABA inhibition is critical for the mechanism initiating puberty onset, as chronic infusion of the GABAA receptor antagonist, bicuculline, significantly increased GnRH release and accelerated the timing of menarche and first ovulation in female rhesus monkeys. Because both GnRH and kisspeptin-54 release is low in the medial basal hypothalamus of prepubertal monkeys, and kisspeptin-10 stimulates GnRH release, we hypothesized that low kisspeptin-54 release prior to puberty onset is, in part, due to a tonic GABA inhibition. To test this hypothesis we examined the effects of bicuculline infusion on kisspeptin-54 release using a microdialysis method. We found that infusion of bicuculline at 1[micro]M dramatically stimulates kisspeptin-54 release in the medial basal hypothalamus of prepubertal monkeys, but had little effect on kisspeptin-54 release in midpubertal monkeys. We further examined whether GABA inhibition of kisspeptin-54 release influences GnRH release. Specifically, we investigated whether bicuculline-induced GnRH release is blocked by the presence of the kisspeptin antagonist, peptide 234. Indeed, we found that inhibiting kisspeptin-54 signaling blocked the bicuculline induced stimulation of GnRH release. The data are interpreted to mean that kisspeptin neurons relay GABA signals to GnRH neurons and that a reduction in tonic GABA inhibition of GnRH release before puberty onset is, at least in part, mediated through kisspeptin neurons.[br][br]Sources of Research Support: This research was supported by NIH grants: R01HD11355 and R01HD15433 (ET) and K99ES020878 (JRK) and made possible to perform by NIH support (P51RR000167, RR15459, and RR020141) to the Wisconsin National Primate Research Center.[br][br]Nothing to Disclose: JRK, KLK, KAG, ET 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1996 201 1669 SUN-686 PO19-01 Sunday 1466 2012


1463 ENDO12L_SUN-687 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) Acetylcholine Activates GnRH Neurons and Sustains Gonadotropin Secretion Via Low-Affinity Nicotinic Receptors in Mice Min Wu, Alexander S Kauffman, Meenakshi Alreja Yale University School of Medicine, New Haven, CT; University of California, San Diego, La Jolla, CA; Yale University School of Medicine, New Haven, CT Activation of GnRH neurons is essential for puberty and reproduction. Kisspeptin, a neuropeptide encoded by Kiss1, is the most potent identified activator of GnRH neurons, though some other neurotransmitters also modulate GnRH neuronal function. In this study, using whole-cell patch-clamp recordings in brain slices prepared from pre- and post-pubertal male and female mice, we have identified another potent activator of GnRH neurons. We show that kisspeptin-activated GnRH neurons are also strongly excited by acetylcholine (ACh), a finding that is consistent with a previous study on immortalized GT1-7 cells. ACh activation of GnRH neurons is mimicked by brief applications of nicotine and by multiple nicotinic receptor agonists but not by muscarine. Nicotinic activation of GnRH neurons is profound and is accompanied by a slow non-desensitizing depolarization (3-40 mV) that is mediated via a direct, TTX-insensitive (n=8) post-synaptic mechanism. The high EC[sub]50[/sub] value for nicotinic activation (17 [mu]M; n=5) suggested involvement of low affinity nicotinic receptors. Accordingly, nicotinic activation, that was blocked by d-tubocurarine (n=5) and mecamylamine (n=5), was not blocked by dihydro-[beta]-erythroidine (n=8), a blocker of high-affinity [beta]2 receptors or by methyllycaconitine (MLA; n=6) and [alpha]-bungarotoxin (n=6), blockers of low-affinity nicotinic [alpha]7 receptors. In contrast, [alpha]-conotoxin AuIB attenuated the nicotinic response, indicating involvement of [alpha]3[beta]4 receptors (n=4). Interestingly, nicotinic activation of GnRH neurons occured both in the absence and in the presence of kisspeptin, suggesting involvement of independent excitatory pathways. In in vivo studies, the nicotine receptor antagonist, mecamylamine, significantly blocked gonadectomy-induced increases in LH secretion in mice of both sexes, suggesting a novel and important physiological role of endogenous ACh and nicotinic receptors in gonadotropin release and reproductive function.[br][br]Sources of Research Support: MH61465, State of Connecticut, Department of Mental Health and Addiction Services and Yale CENTURY/TTURC program awarded to MA. NICHD R01 HD065856, U54 HD012303 awarded to ASK and U54 HD28934 (University of Virginia Ligand Assay Lab).[br][br]Nothing to Disclose: MW, ASK, MA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1946 201 1670 SUN-687 PO19-01 Sunday 1467 2012


1464 ENDO12L_SUN-688 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) NMDA-Induced Luteinizing Hormone Release in Young Female Rats Involves Kisspeptin Dependent and Independent Mechanisms Yan Sun, Jun Shu, Genevieve Neal-Perry Albert Einstein College of Medicine, Bronx, NY; Albert Einstein College of Medicine, Bronx, NY Gonadotropin releasing hormone (GnRH) neurons, key regulators of female reproductive physiology, receive afferent input from a diverse group of neurons. The neuropeptide kisspeptin (Kp) is a potent activator of GnRH neurons and GnRH release. Intracerebroventricular (ICV) infusion of Kp immunoneutralizing antibody or antagonist blocks the LH surge. We previously reported that brain infusion of Kp increased hypothalamic glutamate (Glu) levels in microdialysis samples collected from middle-aged female rats. [bold][italic]We hypothesize that Kp modulates GnRH release, in part, by modulating glutamatergic neurotransmission. [/italic][/bold]To test this hypothesis we determined the effect of ICV P318, a Kp receptor antagonist, on [italic]N[/italic]-methyl-D-aspartate (NMDA; ionotropic Glu receptor agonist) induced LH release (marker of GnRH release). Gonadectomized (OVX) female rats (3-4 mon; n=4-6) were primed with estradiol (E) and progesterone (P) before they received 2 ICV infusions of P318 (10 nmol) or vehicle in the 3[sup]rd[/sup] ventricle (9:30 and 10:30h). NMDA was ICV (2ug/2ul) or intravenously (IV) (15mg/kg) infused at 1100h. Plasma was collected every 10-30 min for 3 hrs and LH levels determined by immunoassay. To determine if P318 affected pituitary LH release, P318 or vehicle was ICV infused in OVX rats primed with E and P and challenged with 25 and 100 ng of IV GnRH peptide. Plasma were collected every 30 min for 4 hrs and effects on LH levels determined by immunoassay. ANOVA, repeated measure ANOVA, and t-test were used for statistical analysis. [bold]Results:[/bold] Both ICV and IV injections of NMDA induced a increase in LH compared to baseline LH release (P[lt]0.05). However females infused with IV NMDA released 50% less LH compared to ICV infused females (P[lt]0.03). ICV P318 caused a 2-fold decrease in total LH release (P[lt]0.05) and caused more than a 50% reduction in peak LH release in females infused with ICV NMDA. In contrast P318 did not affect total or peak LH release in females infused with IV NMDA. P318 did not affect GnRH peptide-induced LH release from the pituitary. [bold]Conclusion:[/bold] NMDA-induced LH release involves Kp sensitive and insensitive mechanisms. NMDA-induced LH release that is [italic]sensitive to Kp[/italic] most likely involve GnRH neurons and their nerve terminals. NMDA-induced LH release that is [italic]insensitive to Kp[/italic] most likely involve GnRH independent effects on pituitary gonadotrophs. These data are consistent with our hypothesis that Kp, in part, increases LH release by modulating NMDA-mediated neurotransmission.[br][br]Sources of Research Support: Eunice Kennedy Shriver NICHD/NIH through cooperative agreement U54 HD058155 as part of the SCCPIR and by Albert Einstein College of Medicine.[br][br]Nothing to Disclose: YS, JS, GN-P 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1329 201 1671 SUN-688 PO19-01 Sunday 1468 2012


1465 ENDO12L_SUN-689 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) Activation of Estrogen Receptor [alpha]-Containing Cells in the Medial Preoptic Area, Arcuate Nucleus and Ventromedial Nucleus of Intact Ewes during the Follicular Phase and Its Alteration after Endotoxin Chrysanthi Fergani, Jean E Routly, Lucy C Pickavance, Robert F Smith, Hilary Dobson University of Michigan, Ann Arbor, MI; University of Liverpool, Liverpool, UK Cells located in the medial preoptic area (mPOA), arcuate nucleus (ARC) and ventromedial nucleus (VMN), possessing estrogen receptor [alpha] (ER[alpha]), mediate estradiol feedback and regulate endocrine and behavioral events during the estrous cycle. The aim of this study was to investigate the location of c-Fos, a marker for activation, in ER[alpha] containing cells, at various times during the follicular phase of intact ewes, and to determine if stress-induced disruption of the LH surge is accompanied by a reduction of the percentage of activated ER[alpha]-cells. Follicular phases of intact ewes were synchronized by withdrawal of progesterone vaginal pessaries. Control animals were killed at 0h, 16h, 31h and 40h (n=5-6/group) after progesterone withdrawal (PW; time zero). At 28h, groups of animals received endotoxin (LPS; 100 ng/kg) and were subsequently killed at 31h or 40h (n=5/group). Hypothalamic sections were immunostained for ER[alpha] and c-Fos. LH surges occurred only in control ewes with an onset at 36.7[plusmn]1.3h after PW: these animals had a marked increase in the percentage of ER[alpha] cells co-expressing c-Fos in the ARC (from 25 to 64%; P[lt]0.05) and mPOA (from 17 to 40%; P[lt]0.05) from 31h after PW and throughout the LH surge, but only during the surge for the VMN (from 32 to 65%; P[lt]0.05). Control animals were then re-grouped according to whether they had exhibited sexual behavior (onset at 28.5[plusmn]2.4h after PW). There was an increase in the percentage of ER[alpha]-containing cells that co-expressed c-Fos in the VMN in animals that had expressed sexual behavior compared to those that had not (from 5 to 57%; P[lt]0.001). At 31h and 40h after PW (i.e., 3h and 12h after treatment, respectively), LPS decreased the percentage of ER[alpha] cells co-expressing c-Fos in the ARC (from 64 and 56% to 12 and 18%, respectively; P[lt]0.05) and mPOA (from 40 and 62% to 11 and 13%, respectively; P[lt]0.05), but there was no change in the VMN, compared to controls. These results suggest that there is a distinct temporal pattern of ER[alpha]-containing cell activation in the hypothalamus during the follicular phase, which begins in the ARC and mPOA 6-7h before the LH surge onset, and extends to the VMN at the time of sexual behavior and the LH surge. This pattern is markedly disrupted by acute LPS administration during the late follicular phase, suggesting that the disruptive effects of LPS are mediated by interruption at the level of the mPOA and ARC prior to the onset of the LH surge.[br][br]Nothing to Disclose: CF, JER, LCP, RFS, HD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1164 201 1672 SUN-689 PO19-01 Sunday 1469 2012


1466 ENDO12L_SUN-690 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) Genetic Mechanisms Mediating Kisspeptin Regulation of GnRH Gene Expression Horacio J Novaira, Doris Fadoju, Sally Radovick The Johns Hopkins University School of Medicine, Baltimore, MD Kisspeptins (KS), products of the KiSS-1 gene, as well as their G-protein coupled receptor 54 (GPR54), have been shown to be key components in the regulation of gonadotropin-releasing hormone (GnRH) secretion in several mammalian species, including humans. In addition, [italic]in vitro [/italic]studies have demonstrated an increase in GnRH gene expression associated with an elevated secretory response to KS administration, suggesting that KS mediates GnRH expression at both the secretory and pretranslational levels. However, the cellular targets and intracellular mechanisms mediating KS effects in the central reproductive axis are unclear. In this study, we define specific regions of the mouse GnRH (mGnRH) promoter; transcription factors (TFs) and chromatin modifications that mediate KS action on target gene expression in GnRH expressing neuronal cell lines (GT1-7 and GN11) and transgenic mice. [italic]In vitro[/italic], transient transfection studies in these cells using sequential deletions of the mGnRH gene promoter fused to the luciferase (LUC) reporter gene demonstrate that KS significantly increases LUC activity when cells are treated with 10[sup]-9[/sup]M KS, thus localizing a kisspeptin-response element (KsRE) between -3446 bp and -2806 bp of the mGnRH gene. [italic]In vivo[/italic], transgenic mice bearing sequential deletions of the mGnRH gene promoter linked to the LUC reporter were used. KS (1nmol via IP) treatment also increased LUC activity in the hypothalamus of male and female mice by 2-fold and again a KsRE was localized between -3446 bp and -2806 bp of the mGnRH gene. Interestingly, this region contains a previously identified GnRH enhancer element (GnRHen). To define the mechanism of regulation, 10[sup]-9[/sup]M KS was first shown to induce nucleosome-depleted DNA within the KsRE in GT1-7 cells by FAIRE assay. DNA sequence analysis revealed a potential binding site for the transcription factor, Otx-2 in the GnRHen, located in the KsRE. KS treatment of GT1-7 and GN11 cells increased mRNA levels of Otx-2 in a time-dependent manner. ChIP assays demonstrated increases of Otx-2 binding after KS treatment to its binding site locate in the KsRE. In conclusion, this work identified elements between -3446 bp and -2806 bp in GnRH-neuronal cell lines and in transgenic mice to play a significant role in positive regulation of GnRH by KS. In addition, we show for the first time that Otx-2 is regulated by KS, and plays a role in mediating the transcriptional response of mGnRH gene.[br][br]Sources of Research Support: This research was supported by the Eunice Kennedy Shriver NICHD/NIH through cooperative agreement [U54 HD 933067 (The Baltimore-Chicago Center for Reproductive Research)] as part of the Specialized Cooperative Centers Program in Reproduction and Infertility Research (SCCPIR), R01 HD370246, and through cooperative agreement [partnership U01HD066432] as part of the Cooperative Partnerships to Promote Workforce Diversity in the Reproductive Sciences Program.[br][br]Nothing to Disclose: HJN, DF, SR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 932 201 1673 SUN-690 PO19-01 Sunday 1470 2012


1467 ENDO12L_SUN-691 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) Deletion of the GnRH Enhancer Element: Effect on Pubertal Onset and Estrous Cycling Sara A DiVall, Bonnie Sklar, Alexia Haralambous, Horacio J Novaira, Andrew Wolfe, Sally Radovick Johns Hopkins University, Baltimore, MD Tissue-specific expression of gonadotropin-releasing hormone (GnRH) is critical for normal pubertal onset and reproductive function. Mice with deletion of an enhancer element in the GnRH promoter from -2806 to -2078bp (GREKO[sup](-/-)[/sup]) have markedly reduced hypothalamic GnRH expression and enhanced ovarian GnRH expression, associated with delay in pubertal onset, higher surge luteinizing hormone levels, shortened estrous cycles, and normal fertility. To determine whether the observed phenotypes were due to decreased hypothalamic versus increased ovarian GnRH expression, orthotopic ovarian transplants were performed. At postnatal day (PND) 23, GREKO[sup](-/-)[/sup] mice donated ovaries to GREKO[sup](+/-)[/sup]and GREKO[sup](-/-)[/sup]received the GREKO[sup](+/-)[/sup]ovaries. PND 23 GREKO[sup](-/-) [/sup]and GREKO[sup](+/-)[/sup] mice that underwent sham surgery or received own ovaries (self-transplanted) served as surgical controls. Age of vaginal opening was not significantly different between sham and self-transplanted groups. As expected, GREKO[sup](-/-) [/sup]surgical controls experienced puberty an average of 5 days later than the GREKO[sup](+/-)[/sup] surgical controls. GREKO[sup](-/-)[/sup] and GREKO[sup](+/-)[/sup] mice with donated transplanted ovaries experienced puberty at an intermediate age, three days later than GREKO[sup](+/-)[/sup] self-transplanted mice. Estrous cycling patterns revealed that GREKO[sup](+/-)[/sup] surgical controls experienced 2.5- 2.8 cycles/21 days and GREKO[sup](-/-) [/sup]surgical controls experienced 2.9-3.5 cycles/21 days consistent with previous observations. GREKO[sup](+/-)[/sup] mice with GREKO[sup](-/-) [/sup]ovaries experienced 2.0-3.0 cycles/21 days, while GREKO[sup](-/-) [/sup]mice with GREKO[sup](+/-)[/sup] ovaries experienced one cycle in 21 days. Differences between groups were not significant. Transplants in adult mice revealed that GREKO[sup](+/-)[/sup] mice with GREKO[sup](-/-)[/sup] ovaries experienced 2-5 cycles/21 days while GREKO[sup](-/-)[/sup] mice with GREKO[sup](+/-)[/sup]ovaries experienced 2-3 cycles/21 days. These results confirm the critical role of the hypothalamic GnRH enhancer element in the control of reproductive cycling and suggest that both hypothalamic and ovarian GnRH expression contribute to pubertal timing in mice. The shortened estrous cycles in GREKO[sup](-/-)[/sup] mice are associated with increased expression of ovarian GnRH.[br][br]Sources of Research Support: NIH/NICHD through cooperative agreement U54HD933067 Baltimore-Chicago Center for Reproductive Research. SD supported in part by NIH/NICHD K08HD056139.[br][br]Nothing to Disclose: SAD, BS, AH, HJN, AW, SR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1339 201 1674 SUN-691 PO19-01 Sunday 1471 2012


1468 ENDO12L_SUN-692 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) Inhibition of Gonadotropin and Estrous Cycles in Hyperprolactinaemic Female Mice Is Restored by Kisspeptin-10 Treatment Charlotte Sonigo, Justine Bouilly, Nadege Carre, Virginie Tolle, Philippe Zizzari, Alain Caraty, Javier Tello, Fabio Simony, Robert P Millar, Jacques Young, Nadine Binart Faculty Medicine Paris-Sud, Le Kremlin-Bic[ecirc]tre, France; Univ Paris Descartes, Paris, France; Univ de Tours, Nouzilly, France; University of Edinburgh, Edinburgh, UK; University Pretoria, Cape Town, South Africa; H[ocirc]pital du Kremlin Bic[ecirc]tre, Le Kremlin Bic[ecirc]tre, France Hyperprolactinemia is the most common cause of hypogonadotropic anovulation (WHO Group I) and represents a major etiology of infertility, with peak prevalence for women aged 25-34 years. In men, it is frequently associated with hypogonadism. It is related to an alteration of pulsatile GnRH secretion. This gonadotropic deficiency has been proposed to result from direct suppression of prolactin (PRL) on GnRH release but its mechanism remains unknown. Because GnRH neurons do not express unequivocally the PRL receptor, and are stimulated by kisspeptin (Kp) neurons which do express PRL receptors, we hypothesized that GnRH deficiency in this condition could be due to a decrease in Kp secretion. To test this we developed and characterized a hyperprolactinemic female mouse model mimicking the human pathology and analyzed the ability of Kp10 i.p. administration to restore gonadotropin secretion and cyclicity.[br]We demonstrated that 28 days administration of PRL by micropumps, significantly inhibited mouse ovarian cyclicity evaluated by vaginal smears, and decrease corpora lutea (CL) number, reflecting ovulation rate impairment. This anovulation was related to a significant downregulation of pituitary LH[beta] and FSH[beta] transcripts indicating a gonadotropin deficiency in this model. Hypothalamic GnRH expression was not altered while there was, a significant decrease of Kp mRNA and peptide (hypothalamic staining) suggesting a role of Kp decrease in GnRH release alteration in hyperprolactinemic female mice. We then demonstrated that Kp-10 administration restored cyclicity, ovulation and pituitary gonadotropin expression in hyperprolactinemic mice. Using hypothalamic explants, we also demonstrated that in vitro decrease of GnRH release induced by PRL was rescued by Kp-10 administration.[br]Together with the recent demonstration that Kp neurons express high levels of PRL receptor, our data suggest that PRL excess acts directly on Kp neurons to suppress Kp secretion and downstream GnRH secretion. Kp neurons appear, therefore, to be the missing link between hyperprolactinemia and GnRH deficiency in mammals.[br][br]Sources of Research Support: Inserm.[br][br]Nothing to Disclose: CS, JB, NC, VT, PZ, AC, JT, FS, RPM, JY, NB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 459 201 1675 SUN-692 PO19-01 Sunday 1472 2012


1469 ENDO12L_SUN-693 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) Continuous Kisspeptin Infusion Restores Pulsatile LH Secretion in Patients with Neurokinin B Signaling Deficiencies Jacques Young, Jyothis T George, Javier A Tello, Bruno Francou, Jerome Bouligand, Anne Guiochon-Mantel, Sylvie Brailly-Tabard, Richard A Anderson, Robert P Millar Universit[eacute] Paris-Sud, Paris, France; (INSERM U693) Institut National de la Sant[eacute] et de la Recherche M[eacute]dicale, Paris, France; H[ocirc]pital Bic[ecirc]tre, Paris, France; University of Edinburgh, Edinburgh, UK; University of Edinburgh, Edinburgh, UK; H[ocirc]pital Bic[ecirc]tre, Paris, France; University of Pretoria, Pretoria, South Africa; Medical Research Council/University of Cape Town, Cape Town, South Africa Pulsatile gonadotropin releasing hormone (GnRH) is a requirement for normal reproductive function. Abnormalities in pulse frequency result in reproductive dysfunction. Kisspeptin and neurokinin B (NKB) arre neuropeptides secreted by the same neuronal population in the ventral hypothalamus, which are crucial central regulators of GnRH and thus gonadotropin secretion. Patients with mutations resulting in loss of signaling by either of these neuroendocrine peptides fail to[br]advance through puberty but the mechanisms mediating this remain unresolved. We show that continuous kisspeptin infusion restores gonadotropin and pulse frequency and amplitude in patients with loss-of-function mutations in neurokinin B [italic](TAC3)[/italic] or its receptor, [italic](TAC3R)[/italic]. Thus indicates that kisspeptin on its own is sufficient to stimulate pulsatile GnRH secretion. Moreover, our findings suggest that NKB action is proximal to kisspeptin in the reproductive neuroendocrine cascade regulating GnRH secretion, and may act as an autocrine modulator of kisspeptin secretion. The ability of continuous kisspeptin infusion to induce pulsatile gonadotropin secretion further indicates that GnRH neurons are able to set up pulsatile secretion in the absence of pulsatile exogenous kisspeptin.[br][br]Sources of Research Support: Medical Research Council(UK) _ Experimental Medicine Grant.[br][br]Nothing to Disclose: JY, JTG, JAT, BF, JB, AG-M, SB-T, RAA, RPM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 196 201 1676 SUN-693 PO19-01 Sunday 1473 2012


1470 ENDO12L_SUN-694 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) Sex- and Age-Dependent Variations in Kisspeptin, Neurokinin B and Dynorphin A Immunoreactivities in the Human Infundibular Nucleus Erik Hrabovszky, Csilla S Molnar, Mate T Sipos, Barbara Vida, Philippe Ciofi, Beata A Borsay, Kalman Racz, Herczeg Laszlo, Stephen R Bloom, Mohammad A Ghatei, Waljit S Dhillo, Zsolt Liposits Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary; Neurocentre Magendie, Bordeaux, France; Faculty of Medicine of the University of Debrecen, Debrecen, Hungary; Hammersmith Hospital, Imperial College London, London, UK [bdquo]KNDy[rdquo] neurons co-expressing kisspeptin (KP), neurokinin B (NKB) and dynorphin A (Dyn A) in the arcuate nucleus of female rodents and sheep have been implicated in the generation of the gonadotropin releasing hormone (GnRH) neurosecretory pulses and the negative feedback effects of sex steroids to the reproductive axis. Here we used immunohistochemistry to address age- and sex-dependent variations of KNDy neuropeptide immunoreactivities in the human infundibular nucleus (Inf). Tissue samples from young and aged men and from postmenopausal women were compared.[br]Robust sex differences could be observed in KP immunoreactivity of aged individuals ([gt]50 years) in that the number of KP-immunoreactive (IR) cell bodies, the density of KP-IR fibers and the incidence of their contacts on GnRH neurons in the Inf were much higher in women than in men. The number of NKB cell bodies was also somewhat higher in women than in men, but the regional density of NKB fibers and the incidence of their appositions onto GnRH cells did not differ significantly.[br]Comparison of samples from males also revealed robust age-dependent changes of KP- and NKB immunoreactivities in that the number of labeled cell bodies, the density of immunoreactive fibers and the incidence of their contacts onto GnRH neurons were several-fold higher in aged vs. young men.[br]In all three models, the number of NKB-IR somata was higher compared with that of KP-IR somata. This difference was particularly robust in young men below 30 years of age where NKB-IR somata outnumbered KP-IR somata five-fold. Interestingly, Dyn A-IR cell bodies and fibers could be visualized in low numbers only in the Inf of any model.[br]The high levels of KP and NKB observed in postmenopausal women in these studies may be explained with the lack of sex steroid negative feedback, whereas testosterone can continue to suppress KP, and to a lesser extent, NKB synthesis in age-matched men. On the other hand, increased KP and NKB synthesis observed in aged men may either reflect a gradual decline in sex steroid levels or a reduced sensitivity of the KNDy neurons and the reproductive axis to the negative feedback effects of sex steroids. Finally, the absence of KP (in particular, in young men) and Dyn A from many NKB neurons and their fibers calls for a more careful use of the [bdquo]KNDy[rdquo] neuron terminology and suggests that the functional importance of these neuropeptides might considerably vary among species, between sexes and at different ages.[br][br]Sources of Research Support: OTKA T73002, K83710 and FP7/2007-2013 (n[deg]245009).[br][br]Nothing to Disclose: EH, CSM, MTS, BV, PC, BAB, KR, HL, SRB, MAG, WSD, ZL 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1195 201 1677 SUN-694 PO19-01 Sunday 1474 2012


1471 ENDO12L_SUN-695 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) Neurokinin B Represses Gonadotropin-Releasing Hormone Transcription by a Fos-Dependent Mechanism Christine A Glidewell-Kenney, Paul P Shao, Pamela L Mellon University of California San Diego, La Jolla, CA Genetic studies identified mutations in neurokinin B (NKB) and the neurokinin 3 receptor (NK3R) in human patients with idiopathic hypogonadotropic hypogonadism (IHH), a disorder characterized by low serum LH and disrupted puberty. Recently, NKB was found to regulate LH by a central mechanism involving the regulation of gonadotropin-releasing hormone (GnRH) secretion. GnRH neurons exist in a highly connected network that allows them to integrate information about the environment, stress and nutritional status. Multiple cell populations within this network were found to express NK3R, including kisspeptin neurons and GnRH neurons themselves. However, technical limitations made it difficult to determine the direct effects of NKB on GnRH neurons in vivo. We found that GT1-7 cells, an in vitro model of the differentiated GnRH neuron, express NK3R and respond to treatment with an agonist, senktide, with an acute increase in GnRH secretion. In contrast, long-term senktide treatment reduced GnRH secretion and also repressed the activity of a rat GnRH reporter, indicating reduced transcription. Using truncations and cis-mutations of the -5Kb rat GnRH regulatory region, we identified the previously characterized neuron-specific enhancer 1 as sufficient for this repression. In this report, we present evidence that senktide represses GnRH transcription by a Fos-dependent mechanism. First, we show that senktide induces c-Fos mRNA. Next, we show that the over-expression of c-Fos and c-Jun (AP-1) represses the activity of a GnRH enhancer 1-luciferase reporter. Using a dominant negative Fos, A-FOS, we show that the repression by senktide requires AP-1 complex formation. Furthermore, using chromatin immunoprecipitation assays, we identify enhanced Fos protein association with enhancer 1 and the GnRH promoter with the induction of Fos by TPA. We identify several putative TPA-responsive element (TRE) half-sites in enhancer 1 and the promoter by sequence analysis. Using electromobility shift assays, we further show Fos protein, including c-Fos, binds directly to the TREs between -1737/-1715 bp and -106/-86. Thus, in addition to directly regulating GnRH secretion, we show that NKB could act on NK3R-expressing GnRH neurons to regulate GnRH transcription by a Fos-dependent mechanism that includes direct binding to novel TRE sites identified in enhancer 1 and the promoter.[br][br]Sources of Research Support: National Institutes of Health (NIH) Grants R01 DK044838 and R01 HD020377 (to P.L.M.) and by National Institute of Child Health and Human Development/NIH through a cooperative agreement (U54 HD012303) as part of the Specialized Cooperative Centers Program in Reproduction and Infertility Research (to P.L.M.). P.L.M. was partially supported by P30 DK063491, P30 CA023100, and P42 ES101337. C.G.K. was partially supported by T32 HD007203, the Hartwell Foundation and T32 DK007044.[br][br]Nothing to Disclose: CAG-K, PPS, PLM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 229 201 1678 SUN-695 PO19-01 Sunday 1475 2012


1472 ENDO12L_SUN-696 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) Neurokinin B and the Control of the Gonadotropic Axis in the Rat: Developmental Changes, Sexual Dimorphism and Regulation by Gonadal Steroids Francisco Ruiz-Pino, Victor M Navarro, A H Bentsen, David Garcia-Galiano, Miguel A Sanchez-Garrido, Donald K Clifton, Robert A Steiner, Leonor Pinilla, J D Mikkelsen, Manuel Tena-Sempere University of C[oacute]rdoba, C[oacute]rdoba, Spain; Brigham and Women[apos]s Hospital and Harvard Medical School, Boston, MA; Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark; University of Washington, Seattle, WA Neurokinin B (NKB), encoded by Tac2 in rodents, and its receptor, NK3R, have recently emerged as important regulators of reproduction; NKB has been proposed to stimulate kisspeptin output onto GnRH neurons. Accordingly, NKB has been shown to induce gonadotropin release in several species; yet, null or even inhibitory effects of NKB on gonadotropin secretion have been also reported. The basis for this discrepant findings, as well as other key aspects of NKB function as master regulator of the gonadotropic axis, remain unfolded. We report here that in the rat, LH secretory responses to the NK3R agonist, senktide, display a salient sexual dimorphism, with persistent stimulation in females, regardless of the stage of postnatal development, and lack of LH responses in males from puberty onwards. Such dimorphism was independent of the circulating levels of sex steroids post-puberty, since testosterone administration to adult females failed to prevent LH responses to senktide, nor was LH responsiveness restored in adult male rats treated with estradiol. Sexual dimorphism was also evident in terms of numbers of NKB-positive neurons in the ARC, which were higher in adult female rats. This may stem from differences in the sex steroid milieu during early periods of brain differentiation, as neonatal exposures to estrogens decreased the number of ARC NKB neurons at later developmental stages. Likewise, neonatal estrogenization resulted also in lower serum LH levels that were normalized by senktide administration. Finally, we document here that the ability of estrogen to inhibit hypothalamic Tac2 expression seems specific of the ARC, since estrogen administration in adulthood increased Tac2 levels in the lateral hypothalamus. Altogether, our present data provide a deeper insight into relevant aspects of NKB function as a major regulator of the gonadotropic axis in the rat, including maturational changes, sexual dimorphism and differential regulation by sex steroids.[br][br]Sources of Research Support: BFU 2011-25021 (Ministerio de Ciencia e Innovaci[oacute]n), NICHD/NIH U54HD12629 and RO1HD049651. Marie Curie OIF within the 7th Framework Program of the UE.[br][br]Nothing to Disclose: FR-P, VMN, AHB, DG-G, MAS-G, DKC, RAS, LP, JDM, MT-S 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1414 201 1679 SUN-696 PO19-01 Sunday 1476 2012


1473 ENDO12L_SUN-697 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) Re-Examination of the Ability of Female GPR54 KO Mice To Generate Preovulatory LH Surges under Multiple Hormonal Paradigms Tal Dror, Alexander S Kauffman University of California, San Diego, La Jolla, CA Kisspeptin is a powerful regulator of GnRH neurons and reproduction. In rodents, it[apos]s been postulated that estrogen-responsive kisspeptin neurons in the anteroventral periventricular nucleus (AVPV) mediate estrogen-mediated positive feedback induction of the preovulatory LH surge. However, conflicting evidence exists regarding the ability of mice lacking GPR54 (aka Kiss1r, the kisspeptin receptor) to display LH surges in response to exogenous hormone treatment. While one study reported no LH surges in GPR54 KO or Kiss1 KO mice, another study reported some LH surges in a different GPR54 KO line. Whether this discrepancy reflects the fact that the two studies used different KO mouse strains and/or different hormonal paradigms to induce LH surges is unknown. Here, we tested multiple hormonal paradigms in one GPR54 KO mouse model to see which paradigms, if any, are able to generate LH surges. First, adult GPR54 KO and WT females were ovariectomized (OVX), given an estradiol (E2) implant, and sacrificed 2 days later in the morning or evening (at lights off). Serum LH levels were very low in all morning animals. All WT females displayed LH surges in the evening, whereas none of the GPR54 KO females had LH surges. The next experiment assessed whether the lack of LH surges in KO mice was due to diminished pituitary responsiveness from a lifetime lack of exposure to GnRH. To test this, new adult females were first primed with injections of GnRH to stimulate the pituitary and then subjected to the same OVX+E2 surge paradigm. All mice of both genotypes responded to GnRH priming with elevated LH secretion. However, only WT mice showed LH surges afterwards with E2 treatment, suggesting that the absent LH surges reflects a lack of GnRH stimulation rather than an unresponsive pituitary. To confirm this, double-label [italic]in situ[/italic] hybridization was used to assess cfos induction in GnRH neurons of E2-treated WT and KO females. A high degree of cfos was detected in GnRH cells of WT females sacrificed in the evening but not the morning, whereas little cfos was expressed in GnRH neurons of KO females at any time. Current experiments are testing an additional hormonal paradigm involving E2 + progesterone. Thus far, our findings conclusively show that WT females display LH surges under multiple hormonal paradigms, whereas GPR54 KO mice do not, supporting the notion that kisspeptin-GPR54 signaling is mandatory for proper LH surge induction.[br][br]Sources of Research Support: NSF grant IOS-1025893; NICHD U54 HD012303.[br][br]Nothing to Disclose: TD, ASK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 900 201 1680 SUN-697 PO19-01 Sunday 1477 2012


1474 ENDO12L_SUN-698 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) The GnRH-Independent Effect of Metastin on Gonadotropin Release and Behavior in Hypogonadal Mice Tomohiro Yonezawa, Ryutaro Ichinose, Shiro Kurusu, Mitsumori Kawaminami Kitasato University, Towada, Japan Metastin, also known as kisspeptin, has been identified as an endogenous ligand of G protein-coupled receptor 54 (GPR54). Its signaling is now established as playing a key role in the neural pathway controlling gonadotropin secretion. Unequivocal data demonstrate that metastin neurons directly stimulate the activity of gonadotropin releasing hormone (GnRH) neurons and that metastin-GPR54 signaling plays a prominent role in puberty onset and ovulation. Further, it seems likely that metastin and GPR54 are involved in regulating the activity of neurons outside the GnRH neuronal network. For example, metastin mRNA expression has been reported in the amygdala and abundant GPR54 expression is observed in multiple areas including thalamus and midbrain. In the present study, we investigated the possibility of GnRH-independent effects of metastin in the brain regarding gonadotropin release and behaviors. Hypogonadal (hpg) mice, which are deficient in GnRH production by natural mutation of GnRH-associated peptide, were utilized. In intact adult female hpg mice, the transcriptional level of metastin in the medial basal hypothalamus was higher than diestrous wild type mice, while the luteinizing hormone (LH) plasma level and mRNA expression in the pituitary were lower. The ability of LH synthesis and release were determined by taking pituitary and blood samples 30 minutes after intracerebroventricular (icv) injection of the biologically active C-terminal decapeptide of metastin (Met-10) or a GnRH agonist (GnRHa, des-Gly(Pro9)-GnRH ethylamide). The central injection of Met-10 into adult female hpg mice did not change the LH mRNA expression in the pituitary or the plasma LH levels, although GnRHa, as expected, increased both of them. Regarding the behavioral analyses, the gonadectomized female hpg mice primed with estrogen benzoate and progesterone were introduced in a test box with a wild intact male, just after icv injection of GnRHa or Met-10. The specific sexual behavior indexes were not significantly altered. However, compared to artificial cerebrospinal fluid-injected mice, the contact number (2-fold, 1.8-fold) and time length of contact (3-fold, 2-fold) were significantly higher in GnRHa and Met-10 injected mice, respectively. These results suggest that metastin neurons have little or no direct effect on synthesis and release of LH, while they influence GnRH-independent pathway(s) promoting contact or sexual behaviors.[br][br]Nothing to Disclose: TY, RI, SK, MK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1810 201 1681 SUN-698 PO19-01 Sunday 1478 2012


1475 ENDO12L_SUN-699 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) Neonatal Exposure to Androgen and Estrogen Alters Gene Expression of Kiss1, Kiss1 Receptor and Androgen Receptor in the Hypothalamus of Female Rat Rodrigo R Marcondes, Katia C Carvalho, Gisele Giannocco, Jose M Soares, Jr, Edmund C Baracat, Gustavo A Maciel Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Universidade Federal de S[atilde]o Paulo [ndash] Escola Paulista de Medicina, S[atilde]o Paulo, Brazil CONTEXT: Polycystic ovary syndrome (PCOS) is the most common endocrine disorder in women in reproductive age. Its etiology remains unknown, but environmental factors, such as exposure to sex hormones in early life, have been related to the origins of PCOS. In animals, androgen excess exposure in early life leads to abnormalities in female endocrinology in adulthood, such as permanent estrous and altered gonadotropin secretion, that resembles PCOS. OBJECTIVE: To determine whether neonatal exposure to testosterone and estrogen alters the expression of genes related to Gnrh pulse control, namely, Kiss1, Kiss1 Receptor and Androgen Receptor. METHODS: Fifteen female rats aged 0-3 days were sorted in three experimental groups according with subcutaneous administration of the following compounds: testosterone propionate (1,25 mg) (testosterone group, TG; n=5), estradiol benzoate (0,5 mg) (estradiol group, EG; n=5) and vehicle (0,1 mL) (control group, CG; n=5). With 90 days of age, the animals were sacrificed and the anterior portion of the hypothalamus was removed. Total RNA was extracted and cDNA synthesis was performed to evaluate the expression of genes GnRH, GnRHR, Kiss1 and Kiss1R using quantitative Real Time PCR. Statistical analysis was done using ANOVA and Tukey[apos]s multiple comparison tests. RESULTS: TG animals have a significant increase in AR expression (Relative expression [RQ] mean[plusmn]SD=1,176[plusmn]0,125) than EG (RQ=1,081[plusmn]0,033) and CG (RQ=1,000[plusmn]0,000) (p[lt]0,01). The Kiss1 gene expression was significantly lower in TG (RQ mean=0,551[plusmn]0,157) and EG (RQ mean=0,317[plusmn]0,051) in comparison to CG (Reference=1,000[plusmn]0,000) (p[lt]0,01). Expression in EG was even lower than TG (P[lt]0,01). Conversely, Kiss1R gene was overexpressed in EG (RQ=1,233[plusmn]0,101) compared CG (Reference=1,000[plusmn]0,000) (p[lt]0,01), but the difference in relation to TG (RQ mean=1,059[plusmn]0,269) was not statistically significant (p[gt]0,05). All animals in the treated group (TG and EG) presented permanent estrous. CONCLUSION: Neonatal exposure to Testosterone or Estradiol alters the gene expression of androgen receptor and Kiss1 gene during adulthood. These findings suggest that early exposure to sex steroids might permanently disrupt estrous cycles and Kiss1 gene plays an important role.[br][br](1)Franks S. Medicine. 2009;37(9):441-44. (2)Dumesic DA et al. Reprod Fertil Develop. 2005;17:349-60. (3)Abbott DH et al. Reprod Biol Endocrinol. 2006;4:17. (4)Krsmanovic LZ et al. Trends Endocrinol Metab. 2009;20:402-8.[br][br]Sources of Research Support: Fundacao de Amparo a Pesquisa do Estado de Sao Paulo - FAPESP - 2010/17417-3.[br][br]Nothing to Disclose: RRM, KCC, GG, JMS, ECB, GAM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 990 201 1682 SUN-699 PO19-01 Sunday 1479 2012


1476 ENDO12L_SUN-700 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) Regulation of [italic]KISS1[/italic] Expression in the Arcuate Nucleus of the Female Rhesus Monkey ([italic]Macaca mulatta[/italic]) by Estradiol and Progesterone Ergul Alcin, Abhiram Sahu, Suresh Ramaswamy, Cynthia L Bethea, Tony M Plant University of Pittsburgh, Pittsburgh, PA; Oregon National Primate Research Center, Beaverton, OR While a major role of ovarian steroid feedback directly at the pituitary is well established to regulate gonadotropin secretion throughout the menstrual cycle of the monkey, the contribution of the hypothalamus in this regard is less clear. Kisspeptin, encoded by [italic]KISS1[/italic], plays an important role in driving GnRH secretion, and kisspeptin neurons in the arcuate nucleus (Arc) are considered to be a site of the negative feedback action of estradiol (E) on LH release in several non-primate species. In this study, we investigated the effect of E and/or progesterone (P) replacement on [italic]KISS1[/italic] mRNA expressing neurons in the Arc of ovariectomized (Ovx) monkeys using ISH with a digoxigenin labeled oligo probe for [italic]KISS1[/italic]. Coronal hypothalamic sections (25 [micro]m) were collected at 250 [micro]m intervals from 4 groups of animals treated with placebo or ovarian hormones at the Oregon National Primate Research Center [Psycopharmacology 160:2171, 2002]. The groups (n=5 each) were treated with either empty [control], E-, P- or E + P-filled Silastic capsules. Steroid replacement produced follicular and luteal phase levels of E (95 + 26 pg/ml) and P (9.6 + 1.1 ng/ml), respectively. In Ovx-control animals, intense hybridization with the antisense [italic]KISS1[/italic] probe was observed in multiple neurons throughout the anterior posterior extent of the Arc. [italic]KISS1[/italic] expression was not observed elsewhere in the hypothalamus. In striking contrast, E treatment for 28 days, alone or in combination with P for the last 14 days, abolished the [italic]KISS1[/italic] signal at all levels of the Arc. Interestingly, P alone resulted in a decrease in both the number of [italic]KISS1[/italic] neurons/section (40[plusmn]8 vs 15[plusmn]3, mean[plusmn]sem, P=[lt]0.05) and the hybridization signal in individual [italic]KISS1[/italic] neurons. In other monkeys with identical treatments, E alone, or in combination with P, inhibited LH secretion. P alone in some animals also appeared to suppress LH, but the overall effect of P was not significant. Together these findings indicate that in the female monkey: 1) the negative feedback action of E on LH secretion involves downregulation of [italic]KISS1[/italic] expression in the Arc and 2) P-induced deceleration of LH pulse frequency during the luteal phase may be mediated by kisspeptin neurons in the Arc. The former suggestion will need to be reconciled with the view that the negative feedback action of E on gonadotropin release is mediated exclusively at the level of the monkey pituitary.[br][br]Sources of Research Support: MH62677 (CLB), RR000163 (ONPRC), U54 HD08610 (TMP), HD013254 (TMP).[br][br]Nothing to Disclose: EA, AS, SR, CLB, TMP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 546 201 1683 SUN-700 PO19-01 Sunday 1480 2012


1477 ENDO12L_SUN-701 POSTER SESSION: Regulation of Kisspeptin [amp] GnRH Neurones (1:30 PM-3:30 PM) The Effects of a Kisspeptin-Cell Specific Deletion of PI3K Catalytic Subunits p110[alpha] and p110[beta] on the Hypothalamic-Pituitary-Gonadal Axis Matthew Beymer, Rohia Aziz, Christian Mayer, Makoto Fukuda, Richard Z Lin, Ulrich Boehm, Maricedes Acosta-Martinez State University of New York (SUNY) Stony Brook, Stony Brook, NY; Institute for Neural Signal Transduction, Hamburg, Germany; University of Texas Southwestern Medical Center, Dallas, TX Kisspeptin, a potent regulator of gonadotropin releasing hormone (GnRH) secretion, is a key mediator of metabolic state and the control of reproduction. A suppression of hypothalamic kisspeptin gene expression is observed in conditions of metabolic imbalance that impair GnRH release such as low nutritional status and diabetes. Leptin regulates hypothalamic kisspeptin mRNA levels and a subset of kisspeptin neurons in the arcuate nucleus of the hypothalamus (ARC), a key region for the control of metabolism and reproduction, express leptin receptors. However, it is unknown if these neurons are also regulated by insulin signaling. Phosphoinositide-3-kinase (PI3K) is a key part of various signaling pathways shared by a number of peripheral metabolic hormones, including insulin and leptin, known to regulate metabolic and reproductive functions. We propose that PI3K signaling in kisspeptin neurons participates in relaying metabolic signals to the GnRH network. To determine if the PI3K-Akt pathway in kisspeptin neurons is activated in response to insulin, FoxO1GFP transgenic mice (with a loxP flanked transcriptional blocker) were bred with transgenic mice expressing Cre recombinase in kisspeptin neurons (FoxO1GFP-Kisspeptin). Mice were implanted with intracerebroventricular cannulas and infused with vehicle or insulin after an overnight fast. The subcellular distribution of FoxO1GFP, cytoplasmic vs. nuclear, was analyzed using immunofluorescence. We found that forty-seven percent of kisspeptin cells in the ARC were responsive to insulin treatment compared to eight percent in saline treated controls. To investigate if PI3K signaling in kisspeptin neurons is required for the normal function of the HPG axis in vivo, mice with a kisspeptin-cell-specific deletion of PI3K catalytic subunits, p110[alpha] and p110[beta], (kisspeptin-Cre-p110[alpha][sup]-/-[/sup]/[beta][sup]-/-[/sup]) were generated. Both male and female mice show normal weight gain over time as well as normal pubertal development. No differences were observed in age at first estrus or in estrus cyclicity between Cre negative and Kisspeptin-Cre-p110[alpha][sup]-/-[/sup]/[beta][sup]-/-[/sup] female mice. Our preliminary data indicates that a subpopulation of kisspeptin neurons respond to insulin stimulated PI3K signaling. Ongoing studies are investigating if the ablation of PI3K catalytic subunits in kisspeptin cells affects gonadotropin levels and fertility in adult mice as well as the ability of insulin to stimulate the PI3K-Akt-FoxO1 pathway.[br][br]Sources of Research Support: The Eunice Kennedy Shriver National Institute of Child and Human Development Grant, 5R00HD055446-04; The Office of the Vice President for Research at SUNY Stony Brook.[br][br]Nothing to Disclose: MB, RA, CM, MF, RZL, UB, MA-M 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1978 201 1684 SUN-701 PO19-01 Sunday 1481 2012


1478 ENDO12L_SUN-702 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) A Case of Gonadotropinoma and High Testosterone Level in a 45-Year-Old Man Mahima Gulati, Robert E Jones University of Utah, Salt Lake City, UT We report the case of a 45 year old man who presented to our hospital[apos]s eye clinic, for [ldquo]photophobia[rdquo] and transient [ldquo]black spots[rdquo] in front of his both eyes. His eye exam revealed significant optic nerve thinning bilaterally; his Humphrey visual field testing showed nearly full field of depression except for sparing homonymously in the right inferior quadrant. His pituitary MRI showed [ldquo]49 X 8 mm pituitary adenoma with avid contrast-enhancement[rdquo]. He denied symptoms of increased muscle mass; scalp balding; acne/changes in facial or body hair shaving patterns; voice changes; scrotal mass; testicular size change; libido changes; gynecomastia; headaches; weight/appetite changes; mood changes; acts/thoughts of aggression; erectile dysfunction; lower urinary tract symptoms. He denied any family history of endocrine tumors. His physical exam was notable for significant visual field deficits on confrontational testing. He had no gynecomastia, and had normal muscle bulk in extremities; normal body and facial hair. Baseline hormonal testing at 2.07 PM showed FSH 35.6 (high); LH 10.8 (high); total testosterone [gt]1500 (high); free testosterone [gt]410(high); high prolactin 99; [alpha] subunit high, ACTH 16, cortisol 7, GH 0.2 (normal); IGF-1: 236 (normal); TSH 3.15 (normal), free T4: 0.7 (normal: 0.8-1.7); high estradiol 109 (normal: 10-42), undetectable beta hCG (tumor marker), normal inhibin A and B. All endocrine labs were measured using ARUP[apos]s quantitative electrochemiluminiscent immunoassay, except free testosterone (calculated from a mathematical expression based on the constant for the binding of testosterone to SHBG); estradiol (quantitative HPLC/tandem mass spectrometry), and inhibin B (ELISA). His pre-op labs showed normal hemoglobin and normal PSA. He underwent trans-sphenoidal resection of his pituitary macroadenoma, and did not develop any complications of DI post-op. His day 1 post-op AM labs were FSH 5.4 (normal); LH: 0.5 (low); total testosterone: 94 (low); free testosterone: 20 (low). He did develop hypocortisolism post-op (AM cortisol was 1.6), and was treated with hydrocortisone and levothyroxine. His tumor pathology showed [ldquo]pituitary adenoma, immunostain negative for prolactin[rdquo], FSH, LH immunohistochemical stain pending. Now 2 weeks post-op, he is doing well. This case highlights the rare scenario of bioactive FSH, LH secretion by a large pituitary tumor, with biochemically elevated free testosterone level.[br][br]Nothing to Disclose: MG, REJ 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2271 202 1685 SUN-702 PO55-01 Sunday 1482 2012


1479 ENDO12L_SUN-703 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) TSH-Secreting Pituitary Incidentaloma in a Patient with Primary Hypothyroidism: Diagnostic and Therapeutic Difficulties Liza C Yambay, Dima Abdelmannan, Baha Arafah UH-Case Medical Center, Cleveland, OH; Louis Stokes Cleveland VA Medical Center, Cleveland, OH Pituitary incidentalomas are present in up to 10% of the population and most are clinically insignificant. TSH secreting adenomas are rare and typically large at presentation. We present the diagnostic and therapeutic difficulties encountered in a patient with longstanding Primary Hypothyroidism who was found to have an incidental pituitary adenoma that proved to be TSH secreting.[br][bold]Clinical Case: [/bold]A 77 year old male with a 26 year history of Primary Hypothyroidism was found to have a 1.7cm pituitary mass abutting the optic chiasm. The Hypothyroidism was treated with 200mcg of thyroxine/day for many years but the dose was increased by his internist 2 years prior to presentation up to 250mcg due to rising serum TSH despite persistently elevated FT4 levels. His exam showed features consistent with hyperthyroidism, gynecomastia and normal eye exam. Laboratory data revealed a TSH of 16.8uIU/mL, FT4 of 2.1ng/dL (n 0.9-1.7), FT3 of 3.5pg/mL (n 2.3-4.2), positive TPO antibodies and an alpha subunit (SU) serum level of 1.5ng/mL (n 0.09-0.76) with alpha SU/TSH molar ratio of 1.0. TSH assay interference was ruled out by repeatedly negative anti-mouse heterophile antibodies. The diagnosis of TSH secreting adenoma in the setting of Hashimoto[apos]s Disease was suspected. A low dose cosyntropin stimulation test revealed a suboptimal cortisol response and hydrocortisone therapy was initiated. Other lab data showed central hypogonadism with increased SHBG and elevated serum prolactin (26-46ng/mL). Given the tumor size, its chiasmal compression and the presence of partial hypopituitarism, he was referred to neurosurgery. He refused surgery and therefore medical therapy with monthly octreotide LAR injections (30mg) was initiated while the thyroxine dose was decreased to 125mcg/day. There was significant decrease in tumor size on repeat MRI after 3 months of therapy. Serum TSH (0.665uIU/mL) and FT4 (1.5ng/dL) became normal with an associated clinical improvement and normalization of SHBG level. Interestingly, the alpha SU/TSH molar ratio increased and remained elevated (2.5-6.7) throughout octreotide therapy. With the decrease in tumor size, improvement of pituitary function was noted such that gonadal, adrenal and prolactin secretion became normal.[br][bold]Conclusion: [/bold]The diagnosis of TSH secreting adenoma can be challenging especially when underlying Primary Hypothyroidism is present. Incidentalomas should be worked up based on the appropriate clinical setting.[br][br]Brucker-Davis et al., J Clin Endocrinol Metab 1999; 84:476-486. Beck-Peccoz et al., Pituitary 2002; 5:83-88. Langlois et al., Thyroid 1996; Aug;6(4):329-35.[br][br]Nothing to Disclose: LCY, DA, BA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1028 202 1686 SUN-703 PO55-01 Sunday 1483 2012


1480 ENDO12L_SUN-704 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) A Rare Case of TSH-Secreting Pituitary Adenoma Nesreen Benhamed, Reem Kheetan, Hani Alkhankan, Omolola Olajide Marshall University, Huntington, WV Introduction:[br]Thyroid stimulating hormone (TSH)-secreting pituitary adenomas are rare and comprise less than 1% of all pituitary tumors.[br]We present an unusual case with pituitary adenoma co-secreting TSH and GH with clinical features of hyperthyroidism and acromegaly.[br]Case presentation:[br]This is a 61 yr old female patient who presented to an Endocrionologist with a history of palpitations, weight loss, sweating for few months. She also complained of enlargement of her hands, increased shoe size, as well as coarse facial features. She has been on Synthroid for few years for elevated levels of TSH.[br]TSH 5.55 (0.4-4.5 uIU/ml), GH 19.4 (0-7ng/ml), prolactin 16.7 (4.8-23.3 ng/ml), alpha subunit 18 ng/ml, free T4 2.88, IGF-1 6166, Free T3 11.1 (2-4.4 pg/ml), LH 27.1 mIU/ml, FSH 61.5 mIU/ml.[br]MRI of the brain showed a 2.1x1.9x2.2 cm pituitary mass with superior extension into the optic chiasm, lateral extension into cavernous sinus and left internal carotid artery.[br]Patient subsequently had a trans-sphenoidal resection of the tumor; the pathology report was consistent with plurihormonal growth hormone secreting adenoma.[br]Discussion:[br]TSH secreting adenomas secrete biologically active thyrotropin in an autonomous fashion. TSH alone is secreted in 71% of TSH secreting adenomas, other pituitary hormones are co-secreted in 29% of cases. The most common co-secreted hormone is the GH. The frequency for growth hormone is 16%, prolactin 11%, FSH and LH 1.4%.[br]The molecular basis of TSH secreting adenoma is still not well known, but molecular analysis of these adenomas show there is some association with over-expression of pit specific transcription factor 1, as well as somatic mutations in thyroid hormone receptor beta gene.[br]The first treatment option in TSH and GH secreting adenomas is surgery.[br]It is generally not possible to excise these lesions completely as they are usually diagnosed at the Macroadenomas. These tumors have fibrotic and invasive properties and only 1/3 of patients are surgically cured. Pituitary radiotherapy and/or medical treatment with somatostatin analogs are two valid alternatives if surgery is contraindicated as well as in the case of surgical failure.[br]Conclusion:[br]In conclusion, TSH secreting pituitary adenomas are rare cause of hyperthyroidism. This Diagnosis should be considered in a patient with a normal or high serum TSH concentration and elevated free thyroid hormones.[br]The initial treatment option for a TSH secreting adenoma is trans-sphenoidal resection.[br][br]1.Thyrotropin-secreting pituitary adenomas: clinical and biochemical heterogeneity. Case reports and follow-up of nine patients.AUGesundheit N, Petrick PA, Nissim M, Dahlberg PA, Doppman JL, Emerson CH, Braverman LE, Oldfield EH, Weintraub BDSOAnn Intern Med. 1989;111(10):827. 2.The changing spectrum of TSH-secreting pituitary adenomas: diagnosis and management in 43 patients.AUSocin HV, Chanson P, Delemer B, Tabarin A, Rohmer V, Mockel J, Stevenaert A, Beckers ASOEur J Endocrinol. 2003;148(4):433.[br][br]Nothing to Disclose: NB, RK, HA, OO 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2011 202 1687 SUN-704 PO55-01 Sunday 1484 2012


1481 ENDO12L_SUN-705 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) An Aggressive Pituitary Tumor Posing Difficulty in Clinical Decision Making Shaghayegh Khayambashi, Varun Mehta, Laura Knecht St Joseph[apos]s Hospital, Phoenix, AZ [bold]Introduction:[/bold][br]TSH-secreting pituitary adenomas are rare, constituting less than 1% of all pituitary adenomas. They are a rare cause of thyrotoxicosis when compared to the more prevalent primary hyperthyroidism. With the introduction of ultrasensitive TSH assays and improved imaging techniques, it is now possible to diagnose TSH-omas at earlier stages and avoid unnecessary and dangerous treatments such as thyroid ablation or thyroidectomy.[br][bold]Case Report:[/bold][br]A 43 year old female presented in Jan, 2011 with symptoms of palpitation, diaphoresis, decreased energy and a 40 Ib weight loss over 3-4 months. Patient was diagnosed with a pituitary macroadenoma measuring 6.7 cm in March, 2008. The tumor involved extra sellar regions and extended to the right middle cranial fossa. She underwent a transsphenoidal tumor resection in March, 2008 and a subsequent right fronto-temporal craniotomy in May, 2008. The tumor could not be completely resected secondary to its anatomical position and the patient received radiation for 6 weeks in May and June, 2009. Her TSH was in the upper limit of normal 4 months after radiation and the MRI showed residual pituitary tumor. Methimazole was started in 2010 by the patient[apos]s PCP in order to maintain a euthyroid state but her TSH continued to increase to 11.14 in Nov, 2011 and she remained hyperthyroid.[br][bold]Discussion:[/bold][br]The majority of patients with TSH-secreting tumors present with macroadenomas, possibly secondary to delay in diagnosis. It is essential to rule out central hyperthyroidism to avoid improper therapy attempts such as thyroid ablation or thyroidectomy. TSH-secreting adenomas should initially be treated surgically. Only one third of these tumours will be cured by surgery alone because most are macroadenomas, which tend to be locally invasive. Radiotherapy is recommended as routine adjunctive therapy when surgery has not been curative. Even after both surgery and radiotherapy only two thirds of tumours will be under control biochemically. Numerous medical therapies have been attempted in the patients who remained hyperthyroid after surgery and radiotherapy. These include dopamine agonists such as cabergoline, bromocriptine and somatostatin analogs such as octreotide. The outcome has been variable for dopamine agonists. Treatment with octerotide, however, has shown a favourable effect of this drug on TSH secretion, thyroid function, and, in rare cases, pituitary adenoma size.[br][br](1) Freda P, Wardlaw S 1999 Diagnosis and Treatment of Pituitary Tumors. J Clin Endocrinol Metab 84: 3859-3866. (2) Kienitz M, Quinkler M, Strasburgery C, Ventz M 2007 Long term management in five cases of TSH-secreting pituitary adenoma: a single center study and review of the literature. European J Endocrinology 157: 39-46. (3) Chanson P, Weintraub B, Harris A 1993 Octreotide therapy for thyroid-stimulating-hormone-secreting pituitary adenomas. Ann Intern Med 119: 236-240. (4) Clarke M, Erickson D, Castro R, Atkinson J 2008 Thyroid-stimulating hormone pituitary adenoma. J Neurosurg 109: 17-22. (5) Socin V, Chanson P, Tabarin A, Rohmer V, Mockel J, Stevenaert A, Beckers A 2003 The changing spectrum of TSH-secreting pituitary adenomas: diagnosis and management in 43 patients. European J of Endocrinology 148: 433-442. (6) Kole M, Goldman J, Rock J 1997 TSH-secreting pituitary adenoma: current management and review. Skull Base Surgery 7: 89-93. (7) Dhillon K, Cohan P, Kelly D, Darwin K, Iyer K, Chopra I 2004 Treatment of hyperthyroidism associated with thyrotropin-secreting pituitary adenomas with iopanoic acid. J Clin Endocrinol Metab 89: 708-711.[br][br]Nothing to Disclose: SK, VM, LK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 231 202 1688 SUN-705 PO55-01 Sunday 1485 2012


1482 ENDO12L_SUN-706 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) The Diverse Presentation and Management of Clinically Active Thyroid-Stimulating Hormone (TSH) Adenomas Alyson Myers, Todd Morgan, Michael J McPhaul University of Texas at Southwestern, Dallas, TX Introduction/Background: Many pituitary adenomas, which immunostain positively for pituitary hormones do not have clinical manifestations as the hormone is often biologically inactive or there is an inability for it to bind to its receptor.(1) In this case series, we demonstrate the variety of presentations of patients with pituitary adenomas that immunostain positively for TSH and are clinically active.[br]Clinical Case: Four patients have been seen in our medical center for central hyperthyroidism (CH) which was caused by clinically-active TSH secreting adenoma. Presenting complaints included headache with visual bilateral worsening visual loss, dizziness, palpitations and loose bowel movements. All patients denied a family history of central hyperthyroidism. The CH work-up involved assessment of thyroid function tests, pituitary axis hormones, [alpha]-subunit, and MRI imaging of the pituitary. Confirmatory testing for [alpha]-subunit:TSH ratio [gt]1 was done in 2 of 4 cases. T3 suppression testing, another confirmatory testing option, was not performed in any of these patients, as they were all either elderly and/or had cardiac complaints. 3 of 4 patients were pre-operatively treated with methimazole and 1 had been treated with levothyroxine after having a previous lobectomy. All four patients were referred for surgery, as MRI confirmed the presence of a pituitary macroadenoma, and all had signs and symptoms of hyperthyroidism. None of the tumors demonstrated cavernous sinus involvement and one had superior displacement of the optic chiasm. Post-operatively 1 patient continued to have signs and symptoms of hyperthyroidism thus requiring the use of Sandostatin LAR[trade] (octreotide). 2 patients were lost to follow-up. All 4 of the tumors had a mitotic index of 0/10 and stained positively for synaptophysin and TSH.[br]Conclusions: Cases of clinically-active TSH adenoma may differ markedly in presentation and the mild elevation of TSH may lead to misdiagnosis.[br][br]1. Ho DM et al. Histopathology. 2001;39(3):310-9.[br][br]Disclosures: MJM: Investigator, Corcept Therapeutics, Novartis Pharmaceuticals. Nothing to Disclose: AM, TM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1034 202 1689 SUN-706 PO55-01 Sunday 1486 2012


1483 ENDO12L_SUN-707 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Acromegaly and Systemic Lupus Erythematosus [mdash] What[apos]s the Connection? Jose Italo Soares Mota, Tania Bulcao Ferraz, Rejane Belchior Macedo, Alan Aguiar Muniz, Samille Frota Coelho, Ivna Aguiar Henriques, Mardonio Oliveira Lima, Paulo Cruz Queiroz, Antonio Gilson Aragao Junior, Jackson Gondim, Dalgimar Bezerra Menezes, Luciano Franco Hospital Geral de Fortaleza, Fortaleza, Brazil; Hospital Geral de Fortaleza, Fortaleza, Brazil; Hospital Geral de Fortaleza, Fortaleza, Brazil; Hospital Geral de Fortaleza, Fortaleza, Brazil Acromegaly is a neuroendocrine disease which a prevalence of 30-40 per million, having a relationship with other co-morbities such as other neoplasms.[br]Co-secreting adenoma of prolactin and GH is usual, and prolactin has a role in modulating the immune response.[br]Here we report a case of a 18 years old female, diagnosed of Acromegaly due an adenoma co-secreting prolactin ang GH, treated by transesphenoidal surgery without cure, followed by octreotide and carbegolin treatment who developed SLE (systemic erithematous lupus) after 2 years of diagnosis.[br]She overt SLE as a cutaneous rash, with photosensitivity, and prednisone 5mg was started with improvement of this cutaneous manifestations. Three months after that she returned to our emergency service complaining of oliguria, foamy urine, anorexia and foot ulcera. The admission exams revealed nephotic proteinuria (2,3g/24h), high titrate of anti-DNA antibody (1:640), and low complement.[br]Cutaneous vasculitis was seen in palms, ankles and soles. To ophthalmoscopic exam (fundoscopy), was found optic nerve atrophy and vasculitis. Pulsed methylprednisolone therapy was started with beneficial effects, and than cyclophosphamide after rule out infection.[br]A MEDLINE search was used to identify relevant articles. No one case of Acromegaly and SLE was find, although was known that prolactin has immunostimulatory properties with elevated levels reports in SLE patients.[br]It[apos]s a rare case, of an auto-immune disease associated with Acromegaly.[br][br]Nothing to Disclose: JISM, TBF, RBM, AAM, SFC, IAH, MOL, PCQ, AGAJ, JG, DBM, LF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1903 202 1690 SUN-707 PO55-01 Sunday 1487 2012


1484 ENDO12L_SUN-708 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Systemic Complications of Acromegaly in Colombia Alin Abreu, Alvaro Rueda Centro M[eacute]dico Imbanaco, Cali, Colombia; Universidad Libre Secci[oacute]nal Cali, Cali, Colombia Acromegaly is a clinical disorder characterized by progressive somatic disfigurement and a wide range of systemic complications mediated by effects of GH and IGF-I hypersecretion. Due it slow clinical progression, diagnosis is often delayed four to ten years or more, increasing morbidity and mortality due to cardiovascular or respiratory disease, metabolic complications or malignancies. The frequency of these complications in patients with Acromegaly in Colombia is scarce. Purpose: to characterize and determine the frequency of the systemic complications of acromegaly in a sample of patients from Cali, Colombia. Methods: Case series. The endocrinology department at Centro M[eacute]dico Imbanaco in Cali, Colombia is a regional referral center for patients with Acromegaly. All cases from patients with Acromegaly consulting to our center were retrieved and reviewed. Patients were called in and systematically evaluated to determine the presence of hypertension, carbohydrate intolerance, diabetes, cardiovascular disease, sleep apnea, and intestinal disorders during their visit. RESULTS: 32 patients were evaluated (59,37% females, 40,63% males; origin: 68,75% urban, 31,25% rural) with a median age 46[plusmn]16,6yrs. At evaluation, the time of disease evolution was 11,40[plusmn]6,12 yrs, 75% had macroadenomas, 21,87% had microadenomas and 3,12% did not have either of them. The most frequent previous treatment was surgery (40,62%), followed by medical 15,62%; 40,62% had no treatment and 3,12% received surgery+medical treatment. Lab values were IGF-I 1039,75[plusmn]482,18ng/ml; FBG 113,06[plusmn]26,16mg/ml, basal GH 16,24[plusmn]11,85ng/ml, 120 min glucemia 168,87[plusmn]46,95mg/ml, 120 min GH 19,63[plusmn]15,33ng/dl, prolactin 89,50[plusmn]202,11ng/ml. Clinical complications present: acromegalic cardiopathy 46,87%; diabetes 25%; HTN 59,37%; arrhythmias 31,25%; sleep apnea 31,25% (severe), 15,62% (moderate), 6,25% (mild); adenomatous polyposis 12,5%, hyperplasic polyps 12,5%,colon cancer 3,12%. Ordered treatment after evaluation: medical 25%, surgery 9,37%; medical+radiosurgery 21,87%; medical+surgery 31,25%; surgery+radiosurgery 3,12; medical+surgery+radiosurgery 9,37%. CONCLUSIONS: Cardiovascular and respiratory complications are frequent among acromegalic patients. The presence of complications in patients with acromegaly should be evaluated at the time of diagnosis y on a regular basis.[br][br]Nothing to Disclose: AA, AR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 199 202 1691 SUN-708 PO55-01 Sunday 1488 2012


1485 ENDO12L_SUN-709 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Vanishing Hypercalciuric Kidney Stone Formation after Treating Underlying Acromegaly Eline S van der Valk, Tom JS Tobe, Aline Stades, Alex F Muller Diakonessenhuis Utrecht, Utrecht, Netherlands; University Medical Center Utrecht, Utrecht, Netherlands A 53-year old male with a medical history of hypertension, type 2 diabetes mellitus, osteo-artritis and obstructive sleep-apnoea, was referred because of recurrent kidney stones. For the past 1,5 years he had excreted approximately one renal stone each month. Other symptoms included: dental problems, an enlarged tongue, thickened skin, increased perspiration and enlarged feet and hands that had developed over the past years. Besides his acromegalic features the physical examination was normal. Analysis of the stones revealed them to be 100% calcium oxalate. Additional investigations showed normal plasma concentrations of calcium, phosphate, parathyroid hormone, vitamin D,. Urinary phosphate [ndash]excretion was normal, but a hypercalciuria (0.4/mM creat) and hyperoxaluria 18.5 microM/mM creat) was identified. There was no evidence of sarcoidosis.[br]Considering the clinical presentation acromegaly was suspected and the IGF-1 level was found to be significantly increased (1). MRI of the brain revealed a pituitary macro-adenoma with supra- and parasellar extension. There was secondary hypogonadism, but pituitary-adrenal and pituitary-thyroid axes were unaffected.[br]He was referred for neurosurgery and prescribed a long-acting somatostatin analogue.After initiation of a somatostatin analog excessive perspiration disappeared, the apnoeas were less frequent and the frequency of hypoglycaemia[apos]s increased. The hypertension was beter controlled. Nine months after diagnosis the patient underwent endoscopic transnasal transsphenoidal selective adenomectomy without complications. After the operation and after cessation of the somatostatin analog his IGF-I was normal and he became normoglycaemic without medication. Calcium excretion normalised and no kidney stones have been passed since then. Interestingly, in this case, an evident direct correlation between calcium excretion and IGF-1 is present. The relationship between acromegy and hypercalciuria is well decribed, nephrolithiasis is however a rare occurrence in patients with acromegaly (2, 3). Several authors have suggested. that the increased calcium excretion may be due to an elevated serum 1,25 (OH)2 D concentration perhaps as a consequence of a direct action of growth hormone on the renal 1 alpha-hydroxylase activity (3, 4).[br]Albeit very rare, in the appropriatie context acromegaly needs to be considered as a treatable cause of nephrolithiasis due to hypercalciuria.[br][br]1. Chanson P, et al. Pituitary tumours: acromegaly. Best Pract Res Clin Endocrinol Metab. 2009 Oct;23(5):555-74. 2. Ezzat S, et al. Biochemical assessment of bone formation and resorption in acromegaly. J Clin Endocrinol Metab. 1993 Jun;76(6):1452-7. 3. Shah R, et al. Acromegaly as a cause of 1,25-dihydroxyvitamin D-dependent hypercalcemia: case reports and review of the literature. Pituitary. 2010 Dec 28. 4. Lund B, et al. Calcium and vitamin D metabolism in acromegaly. Acta Endocrinol (Copenh). 1981 Apr;96(4):444-50.[br][br]Nothing to Disclose: ESvdV, TJST, AS, AFM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1289 202 1692 SUN-709 PO55-01 Sunday 1489 2012


1486 ENDO12L_SUN-710 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Dilated Cardiomyopathy in Two Patients with Acromegaly Veronique Pelletier, Emilie Daoust, Jean-Marie Boutin, Omar Serri Centre Hospitalier de l[apos]Universite de Montreal, Montreal, Canada Patients with acromegaly have a high risk to develop cardiomyopathy, which if left untreated or uncontrolled, ultimately results in systolic heart failure and death. We present here two patients which illustrate different aspects of presentation and potential complications.[br]The first patient is a 66-year-old man who was referred to our pituitary clinic for acromegaly refractory to control by octreotide LAR. The patient presented with severe congestive heart failure (CHF) attributed to ischemic heart disease with no other complaints related to acromegaly. Acromegaly was diagnosed 2 years earlier after an incidental finding of a pituitary macroadenoma on CT-scan and confirmed by high IGF-1 (917 ug/L) and non suppressed GH after OGTT (baseline 10 ug/L; nadir 6 ug/L). After 15 months of octreotide LAR treatment, IGF-1 was only partially responsive (571 ug/L). Echocardiography demonstrated severe diffuse global hypokinesis and a left ventricular ejection fraction (LVEF) reduced to 25%. A myocadial perfusion study showed only a minimal ischemic component to the severe hypokinesis suggesting an important contribution of acromegaly to the cardiomyopathy. The patient was treated with a combination of octreotide LAR and pegvisomant at 30 mg daily. Two months later, GH and IGF-1 decreased to normal levels (1.25 and 173 ug/L respectively). Control echocardiography showed unchanged LVEF at 25 to 30%. Clinically, the patient reported less palpitations and angina, but dyspnea was unchanged.[br]The second patient is a 52-year-old man diagnosed with severe dilated cardiomyopathy (LVEF 20%) of unknown etiology 2 years earlier presented with ventricular tachycardia for which he received defibrillation and hyperthyroidism for which he was referred to our clinic. Acromegaly was suspected because of his enlarged hands and confirmed by high IGF-1 (1314 ug/L) and non suppressed GH after OGTT (baseline 20.75 ug/L; nadir 12.52 ug/L). The CT-scan demonstrated the presence of a microadenoma. Octreotide LAR was started and titrated to a maximal dose of 60 mg once a month, after which pegvisomant 20 mg daily was added. Normal IGF-1 values were attained after 4 months of treatment. Control echocardiogram demonstrated unchanged LVEF and cardiac parameters.[br]In conclusion, these cases emphasize the importance of early diagnosis and treatment of acromegaly in order to reverse the hypertrophy and avoid CHF.[br][br]Nothing to Disclose: VP, ED, J-MB, OS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 735 202 1693 SUN-710 PO55-01 Sunday 1490 2012


1487 ENDO12L_SUN-711 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Acromegaly with Subclinical Pituitary Apoplexy, Cerebrovascular Disease and Left Ventricular Thrombus: A Case Report of Therapeutic Dilemma Chandy Lou Patiag Malong, Erick S Mendoza, Maria Fleurdeliz Goco, Mary Jane Tanchee-Ngo, Edward Gacrama, John Paul Tiopiangco, Lerrie Gutierrez, Leilani Mercado-Asis University of Santo Tomas Hospital, Manila, Philippines; University of Santo Tomas Hospital, Manila, Philippines [bold]INTRODUCTION:[/bold] Acromegaly, an insidious but debilitating disorder of growth hormone hypersecretion, is associated with significant morbidity and mortality.There is 10-year reduction in life expectancy due to cardiovascular and cerebrovascular diseases.Congestive heart failure (CHF), a rare complication of acromegaly occurs in less than 3% of patients.We now report a case complicated by CHF, left ventricular (LV) thrombus, cardioembolic stroke and pituitary apoplexy causing dilemma.[br][bold]CLINICAL CASE:[/bold] A 46-year old Filipino female worker had progressive coarsening of facial features, deepening of voice and enlargement of digits 9 years ago heralded by amenorrhea.Seven years after, she developed hypertension (highest BP 180/100 mmHg) and diabetes (FBS=140 mg/dL).She subsequently experienced easy fatigability, orthopnea and pedal edema. Physical examination findings were consistent with acromegalic features.There were no visual field cuts or neurologic deficits.The cardiac apex beat was sustained and displaced.Growth hormone was non-suppressible ([gt]34.90 ng/ml; NV:[lt]3ng/mL) 1 hour after a 75-gram oral glucose loading.Prolactin dilution assay indicates a stalk effect (38.57ng/mL; NV:4.79-23.3ng/mL) and thyroid function tests were normal (TSH=0.503mIU/L, NV:0.4-4.0mIU/L; FT3=2.74pg/mL, NV:1.5-4.1pg/mL; FT4=1.26ng/dL, NV:0.89-1.76ng/dL).Cranial MRI showed 2.1x3.3x2.4 cm pituitary macroadenoma with patchy central areas of hemorrhage.Incidentally, there was a wedge-shaped infarct with hemorrhagic component at the left frontal cortex.There was no hypotension, headache or blurred vision.Dexamethasone 4mg/IV every 8 hours was immediately started.Echocardiogram demonstrated cardiomyopathic changes with global hypokinesia and severe systolic dysfunction (ejection fraction 22%), with LV thrombus.Anticoagulation, though indicated, was not given due to the pituitary and cerebral hemorrhages.Surgery cannot be offered due to the high risk cardiovascular profile.Patient was started on Bromocriptine, insulin, losartan and maintained on steroids.She was stable upon discharge and on follow-up.[br][bold]CONCLUSION:[/bold] In the Philippines where the recommended somatostatin analogs are not available, bromocriptine may be used as first-line treatment to achieve reduction in growth hormone levels in high risk cardiovascular patient.The risks and benefits of any medications must be emphasized in the presence of conflicting clinical features causing therapeutic dilemma such as in the case reported.[br][br]Nothing to Disclose: CLPM, ESM, MFG, MJT-N, EG, JPT, LG, LM-A 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2196 202 1694 SUN-711 PO55-01 Sunday 1491 2012


1488 ENDO12L_SUN-712 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Unknown Colic Fibrous Tumor in an Acromegalic Patient with McCune-Albright Syndrome and Sarcoidosis Helene Bihan, Abdallah Al Salameh, Zineb Labbi, Christine Lagorce, Philippe Wind, Thomas Aparicio, Regis Cohen, Alain Krivitzky Avicenne Hospital, Bobigny, France; Avicenne Hospital, Bobigny, France; Avicenne Hospital, Bobigny, France; Avicenne Hospital, Bobigny, France; Avicenne Hospital, Bobigny, France Introduction[br]Patients with acromegaly have higher risk of colorectal cancer. Screening is recommended every 5 years. Prevalence of adenomatous, hyperplastic colonic polyps or carcinoma is increased. It is known that in McCune Albright syndrome (MAS), fibrous tumors occur typically in bone and skin. But fibrous mass of colon has never been described.[br]Patient[br]We report the case of a 49 year old woman with MAS and acromegaly. She presented precocious puberty, polycystic ovaries, skin spots and fibrous dysplasia leading to diagnosis of MAS at 21 year old. Genetic testing to identify GNAS mutation was not performed. Acromegaly was diagnosed at 22 year old, without evidenced adenoma. Surgery was not possible (fibrous dystrophy of the skull), but treatment by Somatostatin analogs allowed a correct control of the disease. Last IGF1 level was 257 ng/ml (50-307), with mean GH levels around 2.29 ng/ml ([lt] 2.5). In 2010, mediastinal adenopathy led to diagnosis of sarcoidosis, with a spontaneous favourable outcome.[br]Screening colonoscopy discovered a non symptomatic colic stenosis, with 2 biopsies in favour of colonic carcinoma. Colectomy was performed, and anatomopathological study concluded to a 6.5 cm ulcerous lesion with fibrosis invading the colonic wall, without carcinoma. Retrospectively, a previous polyps biopsy described fibrosis 10 years before.[br]Conclusion[br]This is the first report of an unknown colic fibrosis stenosing mass in a patient with acromegaly, MAS and sarcoidosis. Etiology related to each disease and wether colectomy could have been avoided are in discussion. Genetic analysis for GNAS mutation in colic fibrosis lesion and non affected colon is investigated[sup]1[/sup]. Activating GNAS has been described in colic carcinoma[sup]2[/sup].[br][br]1 Weinstein Lee S et al., Endocrine reviews 2001; 22: 675-705. 2 Zacharin M et al., J Med Genet 2001; 48: 458-461.[br][br]Nothing to Disclose: HB, AAS, ZL, CL, PW, TA, RC, AK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 851 202 1695 SUN-712 PO55-01 Sunday 1492 2012


1489 ENDO12L_SUN-713 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) A Novel Missense Mutation of [italic]AIP[/italic] Gene in a Patient with Octreotide-Resistant Non-Familial Gigantism Hidenori Fukuoka, Genzo Iguchi, Kentaro Suda, Masaaki Yamamoto, Hitoshi Nishizawa, Michiko Takahashi, Susumu Seino, Syozo Yamada, Yutaka Takahashi Kobe University School of Medicine, Kobe, Japan; Kobe University School of Medicine, Kobe, Japan; Kobe University School of Medicine, Kobe, Japan [italic]Background[/italic][br]Mutations in the aryl hydrocarbon receptor-interacting protein ([italic]AIP[/italic]) gene are associated with familial isolated pituitary adenomas (FIPA). Patients with [italic]AIP[/italic] mutations tend to be younger at diagnosis and respond poorly to somatostatin analog therapy.[br][italic]Clinical case[/italic][br]A 18-year-old Japanese male with acromegalic face and tall stature (193.6 cm; [gt] +2SD: 182.3cm) was suspected as gigantism. He has no apparent family history of pituitary diseases. His growth acceleration started since 13 years. He was found to have enlargement of the nose, macroglossia, and large hands and feet. He also showed clinical features of hypogonadism including long arm span (201 cm), low dense pubic hair, and small testis (6 cc). Laboratory tests showed excessive growth hormone secretion; morning GH 23.3 ng/ml, IGF-I 660 ng/ml (normal range, 142 [ndash] 391 ng/ml), nadir GH (75 g OGTT) 12.2 ng/ml and hypogonadotropic hypogonadism; LH [lt] 0.1 mIU/ml, FSH 0.1 mIU/ml, testosterone 0.9 ng/ml. MRI revealed macro pituitary adenoma with cavernous sinus invasion (3.0[times]2.6[times]1.6 cm). After trans-sphenoidal surgery, the patient was treated with Octreotide LAR (20 [ndash] 40 mg/M) for the biochemical control of residual tumor. However, serum GH/IGF-1 levels remained above normal range. DNA testing for [italic]AIP [/italic]gene sequencing of the patient showed a novel heterozygous missense mutation that results a truncated Y261X results in disrupt the C-terminal end that is essential for protein-protein interactions, suggesting a loss of function mutation.[br][italic]Conclusion[/italic][br]We present a sporadic case of gigantism with a novel [italic]AIP[/italic] gene mutation. As previously reported in patients with [italic]AIP[/italic] mutation, this patient was also diagnosed at relative younger age, aggressive tumor, and poor responsiveness to octreotide treatment.[br][br]Nothing to Disclose: HF, GI, KS, MY, HN, MT, SS, SY, YT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2281 202 1696 SUN-713 PO55-01 Sunday 1493 2012


1490 ENDO12L_SUN-714 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Pegvisomant (PEG) as Monotherapy or in Combination with Somatostatin Analogs (SSA) in Acromegalic Patients: SSA Do Not Reduce PEG Doses and Diminish the Advantage on Glucose Metabolism (Data from the German Pegvisomant Observational Study) Michael Droste, Michael Buchfelder, Klaus Mann, Anja Schwanke, Gunter K Stalla Endocrinology, Oldenburg, Germany; University of Erlangen-Nuremberg, Erlangen, Germany; University of Duisburg-Essen, Essen, Germany; Pfizer Germany, Berlin, Germany; Max-Planck Institute of Psychiatry, Munich, Germany More than 80 % of all acromegalic German patients under pegvisomant therapy are included in the The German Pegvisomant Observational Study (GPOS). According to current guidelines and drug registration criteria pegvisomat therapy is appropriate for patients after surgery and/or radiation as well as for patients ineffectively treated with somatostatin analogs (SSA). In total 515 patients were analyzed: 359 patients under a pegvisomant montherapy (MONO) and 155 patients under a combination therapy pegvisomant/somatostatin analogs (COMBI). Clinical data of both groups were comparable. A combination therapy was administered to patients with pituitary tumors showing a chiasm near volume extension or for low- or no responders to SSA monotherapy. The ratio of patients suffering from a diabetes type 2 was 30,5 % in MONO and 26,7 % in COMBI. Diabetic patients under a pegvisomant montherapy showed a reduction of the mean HbA1c of 0.5 % (p [lt] 0,005) during their therapy whereas the change in patients under a combination therapy was no significant (HbA1c 0.181 %, p = 0.263). Both patient groups showed comparable rates of IGF- normalization (72,2 % MONO/71,2 % COMBI). Neverless the period until age and gender specific IGF-I normalisation could be achieved was significantly longer in the COMBI group compared to the MONO group (14 vs 11 months, p [lt] 0.05). The mean daily dose of pegvisomant in IGF-I normalized patients was comparable (16,61 mg MONO/15,91 mg COMBI). However, the comparison of the diabetic patient group to the non-diabetic group showed that the mean daily pegvisomant dose was remarkably (p [lt] 0.01) higher in the diabetes group (18,87 mg vs. 15.50 mg) without reaching a comparable IGF-I normalization rate (64.5 % vs. 75.0 %, p[lt] 0.05). These results indicate that high plasma insulin levels in diabetic patients with acromegaly influence the outcome of pegvisomant treatment which is further supported by the higher daily dose (22.8 mg) of pegvisomant in insulin treated diabetic patients. The known reduction of insulin by somatostatin analogs does not result in a reduced daily dose of pegvisomant under combination therapy. Further interactions of growth hormone and insulin should be taken into consideration.[br][br]Nothing to Disclose: MD, MB, KM, AS, GKS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 689 202 1697 SUN-714 PO55-01 Sunday 1494 2012


1491 ENDO12L_SUN-715 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Psychiatric Manifestations of Cushing Syndrome: A Case Report Rachel Brook, Anthony Heaney UCLA Ronald Reagan Medical Center, Los Angeles, CA; UCLA Ronald Reagan Medical Center, Los Angeles, CA Case Presentation:[br]A 45-year-old female with history of hypertension, type 2 DM, and dyslipidemia, was brought to the ED by family for worsening depressive symptoms and refusal to eat; she was subsequently admitted to the psychiatric ICU. Further enquiry revealed the patient had a 6-month history of worsening lower extremity weakness, [quot]sad mood,[quot] and a 50-pound weight loss. In recent months she had undergone extensive multispecialty evaluations for recurrent lower extremity DVTs, nephrolithiasis, rosacea, polycythemia, hirsutism, and oligomenorrhea; she had also undergone liposuction of a dorsal cervical fat pad, right quadriceps EMG and muscle biopsy, and investigation of spontaneous hip fracture. Prior urinary free cortisol was 99.5 ug/dL.[br]On examination, the patient had marked psychomotor retardation, paranoid thoughts, plethoric and rounded facies, hirsutism, truncal obesity, and proximal muscle weakness. Evening serum cortisol was 32 ug/dl; 8 AM serum cortisol did not suppress following 1mg dexamethasone (it was 36). Plasma ACTH was 64 pg/ml and pituitary MRI showed a 7mm hypoenhancing mass keeping with Cushing[apos]s disease. Endoscopic transsphenoidal pituitary surgery was performed; a discrete adenoma was resected and post-op cortisol fell to 0.8ug/dl by 18 hours after surgery. Pathology confirmed a corticotroph pituitary tumor. 6-months post-op the patient had improved and was eating, but she remained severely depressed.[br]Discussion:[br]This case demonstrates the challenge of establishing the diagnosis of Cushing[apos]s syndrome (CS) in a severely psychiatrically ill patient. No single test clearly distinguishes CS and so-called pseudo-Cushing[apos]s (PC) states, such as psychiatric disorders. The combined low dose dexamethasone-CRH test has been reported to be the most helpful as PC patients exhibit a blunted cortisol response following CRH stimulation. [br]This case reminds us that despite resolution of hypercortisolism, 24% of patients still show psychiatric symptoms 12 months after remission with many not ever regaining pre-disease psychologic function (1). In primates high cortisol levels have been associated with neurodegeneration particularly in hippocampal CA1 neurons (2,3). Human studies show structural changes on MR that do not fully resolve despite disease control (4). Prior studies inversely correlate psychological recovery with pre-treatment cortisol levels, although the relationship between hypercortisolemia and affective disorders is complex (5).[br][br](1) Dorn et al. [quot]Subtle Cognitive Impairments in Patients with Long Term Cure of Cushing[apos]s Disease.[quot] Clinical Endocrinology. 2006;64: 314-318. (2) De Kloet et al. [quot]Brain Corticosteroid Receptor Balance in Health and Disease.[quot] Endocrine Reviews. 1998; 19(3):269-301. (3) Patel, PD et al. [quot]Stress-induced changes in the Corticosteroid Receptors in Primate Hippocampus and Prefrontal Cortex.[quot] Psychoneuroendocrinology. 2008 Apr; 33(3):360-7. (4) Starkman, et al. [quot]Decrease in Cortisol Reverses Human Hippocampal Atrophy Following Treatment of Cushing[apos]s Disease.[quot] Biological Psychiatry. 1999; 46 (12): 1595-1602. (5) Olkowitz, O et al. [quot]Glucocorticoids: Mood, Memory, and Mechanisms.[quot] NY Academy of Sciences. 2009; 1179: 19-40.[br][br]Nothing to Disclose: RB, AH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1288 202 1698 SUN-715 PO55-01 Sunday 1495 2012


1492 ENDO12L_SUN-716 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) A Pituitary Mass in a Patient with Cushing Syndrome Catarina Moniz, Joao Anselmo, Joana Boleo, Isabel Sousa, Rui Cesar Hospital Divino Espirito Santo, Ponta Delgada, Portugal; Hospital Garcia de Orta, Almada, Portugal; Hospital Egas Moniz, Lisboa, Portugal An ACTH dependent Cushing syndrome in a patient with a pituitary mass is almost pathognomonic of a corticothropic adenoma. Herein we provide evidence that this is not always the case.[br]A 48-years-old man started complaining of blurred vision and frontal headaches. He was known to suffer of type 2 diabetes and arterial hypertension for several years. A Magnetic Resonance Imaging (MRI) of the brain showed a large heterogeneous pituitary mass occupying the right side of the sella turcica with 25 mm. His blood pressure was 170/100 mmHg. Physical examination was otherwise unremarkable. The laboratorial workout revealed haemoglobin of 17.2 g/dl, HbA1c 9.3% (3.8-5.8); Serum cortisol was 27.07 [mu]g/dl in the morning and 27.0 [mu]g/dl late afternoon; ACTH was 86.4 and 82.6 pg/ml, respectively. Urinary cortical was 526.1 [mu]g/24 hours (28-213). Abnormal 1 mg dexamethasone overnight test (26[mu]g/dl, n[lt]5.0); After 2 mg each 6 hour for 48 hours, urinary cortisol was 141 [mu]g/24 hours (74% suppression). Thyroid function, LH, FSH, PRL and IGF-1 were within the normal range. Testosterone was 382.6[mu]g/dl (241-820).[br]The patient was submitted to transphenoidal surgery with partial remotion of the pituitary mass. The pathologic examination revealed an adenocarcinoma of the sphenoid sinus with low grade malignancy, suggestive of carcinoid but ACTH negative. During the following weeks, normalization of the glycaemia was noticed and the blood pressure returned to normal levels. Therapeutics for hypertension and diabetes were discontinued. Eight months later, hypercortisolism recurred and urinary cortisol was found to be 423 [mu]g/24 hours (28-213). An Octreoscan for somatostanin receptors showed an intense fixation at the sphenoid sinus; CRH level was 73 pg/ml (n: 3.0-7.8); Chromogranin was 124 pg/ml ([lt]74) and urinary 5[apos]indolacethic acid was 16 mg/24 hours (2.0-6.0). A second transphenoidal surgery was carried out. The mass was not attached to the bone. Immunostaining of the tissue removed was positive for chromogranin, synaptophysin and CRH. ACTH and other hypophyseal hormones were not detected.[br]Ectopic production of CRH has been found in the context of different malignant tumors including carcinoid. On the other hand, carcinoid tumors of the sphenoid sinus have been described, but to the best of our knowledge this is the first time that an ectopic Cushing syndrome is dependent on the overproduction of CRH by a carcinoid tumor of the sphenoid sinus.[br][br]Nothing to Disclose: CM, JA, JB, IS, RC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1123 202 1699 SUN-716 PO55-01 Sunday 1496 2012


1493 ENDO12L_SUN-717 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Successful Diagnosis and Treatment of Cushing Disease Due to an Infrasellar ACTH-Producing Adenoma Rebecca Locher, Stefan Bilz, Jean-Yves Fournier, Christian Fretz, Michael Brandle Cantonal Hospital of St Gallen, St Gallen, Switzerland; Cantonal Hospital of St Gallen, St Gallen, Switzerland; Cantonal Hospital of St Gallen, St Gallen, Switzerland [bold]Background[/bold][br]Pituitary adenomas account for the vast majority of cases of endogenous Cushing[apos]s syndrome and may not be detectable by current imaging procedures in up to 50%. Therefore, bilateral inferior petrosal sinus sampling (BIPSS) has been recommended as the standard procedure to differentiate Cushing[apos]s disease from ectopic ACTH-production and guide transsphenoidal surgery.[br][bold]Case Report [/bold][br]In a 72 year old woman with typical cushingoid features the diagnosis of ACTH-dependent Cushing[apos]s syndrome was made by elevated ACTH-concentrations, after endogenous hypercortisolism had been repeatedly confirmed. The MRI (3Tesla) of the sellar region demonstrated a normal pituitary gland as well as an infrasellar 11x7 mm hypointense lesion with delayed and slightly reduced contrast enhancement in the clivus. CT-scan revealed minimal erosion of the sellar floor and faint osteolytic changes in the clivus. BIPSS demonstrated an increased central-to peripheral ACTH-ratio of 3.7 before and 19 after CRH-stimulation, indicating a pituitary ACTH-source. Since a non-detectable pituitary microadenoma could not be ruled out and the etiology of the infrasellar lesion remained ambiguous, a transsphenoidal biopsy was performed revealing a bone resembling tumor intraoperatively. Histologic work-up showed pituitary tissue diffusely positive for ACTH on immunostaining, indicating an infrasellar ACTH-producing adenoma. Invasion of the surrounding osseous structures by the adenoma was visible during the subsequent endoscopic transsphenoidal surgery which succeeded to remove the tumor using a diamond burr. On follow-up a delayed decline of serum cortisol levels reaching a nadir (5ug/dl) on postop day 5 along with clinical signs of adrenal insufficiency indicated successful tumor removal. One year postoperatively normal clinical and biochemical studies confirmed ongoing remission of Cushing[apos]s disease.[br][bold]Conclusion[/bold][br]Extrasellar ACTH-producing adenomas are a very rare cause of Cushing[apos]s disease, may present as micro- and macroadenomas and have been described to occur in the cavernous and sphenoid sinus. The diagnosis should be considered in the work-up of ACTH-dependent hypercortisolism when an extrasellar mass is identified and BIPSS indicates central ACTH-oversecretion. If accessible, transsphenoidal surgery is the treament of choice and may offer long-term cure. However, specific features, such as invasion of osseous structures may complicate resection and should be considered preoperatively.[br][br]Nothing to Disclose: RL, SB, J-YF, CF, MB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1817 202 1700 SUN-717 PO55-01 Sunday 1497 2012


1494 ENDO12L_SUN-718 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Prolactin in Inferior Petrosal Sinus Sampling (IPSS) [mdash] The Importance of Having a Surrogate Pituitary Marker Nisha Acharya, Laurence Kennedy, Robert Weil, Ferdinand Hui Cleveland Clinic, Cleveland, OH; Cleveland Clinic, Cleveland, OH; Cleveland Clinic, Cleveland, OH [underline]Background[/underline]: Once a diagnosis of ACTH-dependent Cushing[apos]s syndrome (CS) is made IPSS is the most reliable way to distinguish pituitary from ectopic ACTH production. Misleading results may occur if ACTH levels in the IPSS do not truly reflect pituitary ACTH secretion.[br][underline]Case report - initial presentation to local hospital[/underline]: A 56 year old man had classic symptoms and signs of CS for 2 years. ACTH-dependent CS was confirmed by standard tests. No pituitary adenoma was seen on MRI. Serum potassium was normal. IPSS, without concurrent prolactin (PRL) measurements, gave baseline peripheral ACTH (pg/mL) 20 with inferior petrosal sinus (IPS) levels of 18 left (L) and 17 right (R). Maximal IPS level was 31(R) compared with 15 in peripheral (P) blood 15 min after CRH. A diagnosis of CS due to ectopic ACTH was made; extensive investigation failed to reveal a source. He was treated with ketoconazole, and later with metyrapone.[br][underline]In our instituion[/underline]: He sought a second opinion about the diagnosis. The [ldquo]flavor[rdquo] of the case, plus review of the investigations, suggested pituitary-dependent CS with false negative IPSS. We recommended repeat IPSS with concomitant PRL and ACTH measurements. PRL (ng/mL) levels in (R) IPS were consistently [gt]2 times (P), but PRL level in (L) IPS and (P) were similar. Basal ACTH (pg/mL) was - (R) IPS 25, (L) IPS 11, and (P) 10. Maximal ACTH post CRH (5 min) was - (R) IPS 284, (L) IPS 17, (P) 14.[br]These results suggest the original IPSS sampling was invalid on both sides. In the repeat IPSS, since PRL levels in (L) IPS are similar to (P), the (L) sampling does not accurately reflect pituitary venous effluent, but on (R), since PRL was consistently [gt]2 times (P), the samples are valid, and ACTH levels, both basal and post CRH, suggest a pituitary source. At transsphenoidal surgery a 0.5mm diameter ACTH-staining microadenoma was removed; post-op serum cortisol was 1.7 mcg/dL and ACTH 5 pg/mL (8-42) in 48h.[br][underline]Conclusion[/underline]: This case illustrates the need to use a surrogate marker, like PRL, to confirm the validity of sampling technique during IPSS, and highlights the importance of interpreting lab results in the context of clinical judgment, especially when there is a conflict between the two.[br][br]Nothing to Disclose: NA, LK, RW, FH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1904 202 1701 SUN-718 PO55-01 Sunday 1498 2012


1495 ENDO12L_SUN-719 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Long-Term Remission of Pulmonary Alveolar Proteinosis after Successful Surgical Resection of a Cushing Disease Pituitary Adenoma Takehiro Kato, Tetsuya Suwa, Reiko Tomita, Takuo Hirota, Katsumi Iizuka, Yukio Horikawa, Jun Takeda Graduate School of Medicine, Gifu University, Gifu, Japan [bold]Background:[/bold] Pulmonary alveolar proteinosis (PAP) is a rare infiltrative pulmonary disease characterized by the alveolar accumulation of surfactant protein. The vast majority of PAP is acquired type, which is due to autoimmunity to the hematopoietic growth factor GM-CSF. Cushing[apos]s syndrome (CS) is considered to induce abnormalities of immune function. Case reports have reported alterations of immunological status after treatment of CS (1).[br]Clinical case: A 56-yr-old woman diagnosed with severe PAP was referred to our hospital to undergo whole lung lavage (WLL). On admission, she was in respiratory failure and was severely hypoxemic despite breathing high-flow oxygen via intubation. GM-CSF autoantibody was positive in both bronchoalveolar lavage and serum. She also exhibited signs and symptoms of hypercortisolism including hypertension, diabetes and central obesity. Initial tests were consistent with ACTH-dependent CS: elevated 24hr urinary cortisol secretion (488 nmol/24hr, 30.9[lt]n[lt]220 nmol/24hr), abnormal 1 mg dexamethason overnight test (cortisol after 1 mg dex 753 nmol/l, n[lt]50 nmol/l), elevated midnight serum cortisol (833.2 nmol/l, n[lt]140 nmol/l), and elevated ACTH-concentrations (32.6 pmol/l, 1.59[lt]n[lt]13.9 pmol/l). MRI showed a 3 mm nodule, which we supposed to be an adenoma, in the right lobe of the pituitary, and a 15 mm cystic lesion on the left lobe. A cavernous sinus venous sampling study revealed elevated ACTH levels, which were higher in the right side in accord with the radiological findings. Response of ACTH to CRH was exaggerated but no paradoxical response to TRH, GRH or LH-RH was observed.[br]Transshenoidal adenomectomy was performed following serial WLL. Successful removal of the ACTH-secreting adenoma was confirmed by postsurgical hypoadrenalism (serum cortisol at 8AM 69.0 nmol/l, 110[lt]n[lt]505). Although WLL was required monthly due to reaccumulation of protein before tumor resection, the pateint has needed only one WLL since the surgery to date. Serum surfactant proteins and GM-CSF autoantibody have remained lower, accompanied by a remarkable improvement of chest radiograph. Hypertension and diabetes were improved as well. Regarding the basis of the cytokine profile, a shift towards T helper 2 responses was observed with the CD19+ cell increase.[br]Conclusion: This is the first reported case of acquired PAP at least in part induced by Cushing[apos]s disease.[br][br](1) Takasu N et al. Exacerbation of autoimmune thyroid dysfunction after unilateral adrenalectomy in patients with Cushing[apos]s syndrome due to an adrenocortical adenoma. N Engl J Med. 1990;322:1708-12.[br][br]Nothing to Disclose: TK, TS, RT, TH, KI, YH, JT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1384 202 1702 SUN-719 PO55-01 Sunday 1499 2012


1496 ENDO12L_SUN-720 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Pediatric Refractory Cushing Disease (CD) Due to ACTH-Secreting Pituitary Macroadenoma Nipapat Visavachaipan, Bonita H Franklin, Constantine A Stratakis, Michael P Wajnrajch, Brenda Kohn New York University School of Medicine, New York, NY; National Institute of Health, Bethesda, MD; Pfizer Global Pharmaceuticals, New York, NY [bold]Background:[/bold] ACTH-secreting macroadenomas account for 4-10% of Cushing[apos]s disease[1]; remission rates in patients with ACTH macroadenoma undergoing surgery are low([lt]65% in most series)[2][br][bold]Case Report:[/bold] A 16 year old female was evaluated for obesity(BMI [gt] 97th percentile), hypertension, and oligomenorrhea. Cushing[apos]s syndrome was confirmed with elevated urinary cortisol level at 628.7[micro]g/24 hr(42-218), elevated midnight serum cortisol (22[micro]g/dl)[3], and no suppression of cortisol(21.8[micro]g/dl) after 1-mg overnight dexamethasone suppression test. MRI brain showed an intra- and suprasellar mass consistent with a pituitary macroadenoma(1.3x1.6x1cm); MRI of the abdomen revealed no adrenal mass. Transphenoidal surgery(TSS) was performed. Histopathology confirmed tissue diffusely positive for ACTH immunostaining, consistent with an ACTH-secreting adenoma. MRI brain post-op showed no evidence of residual pituitary tumor. Prednisone was administered for 5 days post-op due to hypotension presumed resulting from cortisol withdrawal. Cortisol level on post-op day 5 was 2.2[micro]g/dl. Morning cortisol levels ranged between 8.3-15.2 [micro]g/dl(6-21), and 24-hr urinary free cortisol levels(UFC) ranged between 3.8-4.8 [micro]g/24hr(3-55). Despite gradually increasing 24-hr UFC, sequential MRI brain revealed no evidence of tumor. Twelve months after surgery, relapsed CD was confirmed with elevated UFC 65 [micro]g/24hr. Fifteen months after surgery, UFC was elevated at 239 [micro]g/24hr. MRI revealed tumor in the pituitary floor with involvement of both lobes. Patient underwent a second TSS, however tumor was incompletely resected due to bilateral cavernous sinus invasion. Cortisol level at 8 PM on post-op day 4 was 25.6[micro]g/dl. MRI brain confirmed residual tumor and stereotactic gamma knife radiosurgery was planned.[br][bold]Discussion:[/bold] Current literature in CD suggests that serum cortisol levels below 2[micro]g/dl on day 3-5 post TSS predict long term remission, while levels of 2-4.9[micro]g/dl warrant follow-up but no immediate intervention.[4] Since our patient was on prednisone post TSS, the cortisol level of 2.2[micro]g/dl on post-op day 5 cannot be definitively relied upon for its predictive value. In children with CD, low UFCs after TSS are not good predictors of sustained remission.[5] Since surgery is less often curative in CD patients with ACTH-secreting macroadenoma and since no single cortisol cut-off value excludes all patients with recurrence, close monitoring is warranted in all patients with pituitary ACTH macroadenoma.[br][br][1] Storr HL et al., Eur J Endocrinol. 2011 May;164(5):667-74. [2] Biller BM et al., J Clin Endocrinol Metab. 2008 Jul;93(7):2454-62. [3] Batista DL et al., Pediatrics 2007 Sep;120(3):e575-86. [4] Lindsay JR et al., J Clin Endocrinol Metab. 2011 Jul;96(7):2057-64. [5] Batista DL et al., J Clin Endocrinol Metab. 2009 Aug;94(8):2757-65.[br][br]Nothing to Disclose: NV, BHF, CAS, MPW, BK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1826 202 1703 SUN-720 PO55-01 Sunday 1500 2012


1497 ENDO12L_SUN-721 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Nelson Syndrome with No Radiological Evidence of Pituitary Tumor Fleurgin Rochelin, Bhanu Iyer, Kazeem A Saka New York Methodist Hospital, Brooklyn, NY; ELK Medical, Brooklyn, NY [italic]O[bold]bjective[/bold][/italic]:-To report a case of Nelson[apos]s syndrome with no radiological evidence of Pitutary tumor.[br][bold][italic]Methods[/italic][/bold]:- We present a case report of Nelson[apos]s syndrome with clinical, laboratory, and radiological features.[br][bold][italic]Results[/italic][/bold]:-A 56 year old African American female was referred to our facility 4 years ago for c/o hyperpigmentation since 2001. In 1978, patient was diagnosed with Cushing syndrome and underwent transsphenoidal surgery for pituitary adenoma. One year after surgery, since there was no improvement in her symptoms she underwent bilateral adrenalectomy. She did not receive Pitutary radiation before or after adrenalectomy. She was placed on replacement doses of cortisone and flurdocortisone. She denied any headaches, double vision or galactorrhea. Physical exam showed generalized hyperpigmentation of the skin including that of the oral mucosa. Ophthalmologic examination showed no visual field defects. ACTH was noted to be 831pg/ml. Thyroid function, Prolactin, and Igf-1 were noted to be within normal limits. Pituitary/brain MRI repetitively showed no pituitary enlargement, no adenoma. Our patient awaits Ct Scan of Head and Neck and Petrosal Sinus sampling to identify a possible ectopic source for ACTH.[br][bold][italic]Conclusion[/italic][/bold]:-Nelson and his colleagues made the original description of of a pitutary macroadenoma and high plasma ACTH, in a patient with Cushings syndrome who had undergone bilateral adrenalectomy, in 1958(2). Since then many case reports and case series about Nelson[apos]s syndrome have been published, but there is no formal consensus of what defines Nelson[apos]s syndrome. Some will define Nelson[apos]s syndrome according to the classical description with an evolving pituitary mass, whereas others will rely on increasing plasma ACTH levels, even in the absence of a clear pituitary lesion on MRI (2).[br]About 8 to 30% of patients with Cushings disease after bilateral adrenalectomy develop Nelson[apos]s syndrome, anywhere from few months to 24 yrs after adrenalectomy(4). Patients may present with the physical and hormonal consequences of an expanding pituitary mass and, or the effects of ACTH hypersecretion as in or case.[br]In our patient despite several MRI of the brain we are unable to locate the source of pituitary adenoma. There are at least 40 cases where an ectopic pituitary adenoma in Nelson[apos]s syndrome have been reported(1). Most of such tumors have been extracranial and located in the sphenoid bone, sinus, nasal cavity, nasopharynx, clivus and petrous temporal bone(1).[br][br](1) Tucci JR et al;Endocrine Practice 1999;Vol 5 No.3:129-131. (2) Munir A et ak; Arq Bras Endocrino Metab 2007;51/8:1392-1396. (3)V J Moyes., Eur J Endocrinol 2009;160:115-119. (4)Sara Fazio, NEJM december 30th,2011. (5)L kemink, G Pieters, Journal of endocrino metab, 1994 vol79, No 3, 887-889. (6)E Francisco,I Ruiz, The Journal of Larygology and Otology 1997,l11:567-567. (7)T Barber, E Adams, EUR J Endocrinology,2010,163:495-507. (8)G Assie, H bahurel,Journal of endo and metab jan 2007, 92:172-179. (9)R Barabash,F.G Moreno -Suarez, Actas Dermo-sifilograficas, 2010,101:76-80. (10)T yamayi, M Ishibashi, Journal of endo and metab, 1984, 58:790-195. (11)P Kumar,K Simcic, Endocrine Practice, 2008, 14:652-653. (12)V J Moyes, G Alusi,Eur J Endocrinol, 2008, 160:115-119.[br][br]Nothing to Disclose: FR, BI, KAS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1915 202 1704 SUN-721 PO55-01 Sunday 1501 2012


1498 ENDO12L_SUN-722 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Case Report: Silent Corticotroph Adenoma Progressing to Cushing Disease Then to Pituitary Carcinoma with Dramatic Treatment Response Gal Bordo, Huy Duong, Nancy McLaughlin, Robert Cornell, James W Findling, Daniel F Kelly John Wayne Cancer Institute, Santa Monica, CA; University of California Los Angles, Los Angeles, CA; Medical College of Wisconsin, Menomonee Falls, WI; Medical College of Wisconsin, Menomonee Falls, WI Pituitary carcinomas are rare neuro-endocrine tumors with local parasellar invasion and distant metastasis. Herein we report a case of a 54 year old man diagnosed with a silent corticotroph adenoma 14 years ago who developed Cushing[apos]s disease and then progressed to pituitary carcinoma. In 1998, the patient had an evaluation for sinusitis, revealing a sellar mass; endocrine work-up showed elevated ACTH levels between 100-150 pg/mL without biochemical hypercortisolemia. By 2004 he developed stigmata of Cushing[apos]s disease with elevated 24 hour UFC and ACTH levels of 300 pg/mL. He had transsphenoidal subtotal removal of an invasive adenoma. By June 2009 after a steady increase in ACTH and cortisol levels and further tumor growth, he had repeat transsphenoidal surgery. In July 2010 with further tumor growth (40 x 32 mm) and worsening effects of hypercortisolemia, he was referred to our group and had endonasal endoscopic tumor debulking; ACTH levels decreased from 317 pg/ml to 73-137 pg/ml in the post-operative period. Pathology showed ACTH immunostaining and an increased Ki-67 of 5-7%. In September 2010, he began temozolamide therapy, however, over the next several months, his overall condition deteriorated. By May 2011 he had a marked tumor progression with further growth in the sella, clivus, left cavernous sinus and left middle fossa. He then had a two-stage tumor debulking with a left transcranial approach and redo endonasal endoscopic approach. Pathology showed ACTH immunostaining but also marked tumor progression with Ki-67 ranging from 80-90%. Whole body PET-CT showed a liver lesion which was biopsied in June 2011 confirming ACTH-staining pituitary carcinoma. He then began treatment with Cisplatin and Etoposide. By December 2011, both left and right eye vision had also improved significantly although his left sixth nerve palsy persists. His most recent brain and pituitary MRI in January 2012 shows almost complete resolution of the large residual skull base and left middle fossa mass and his body imaging shows the liver lesion has almost completely resolved with no new lesions seen; his ACTH levels have fallen from [gt]350 pg/ml to [lt] 15 pg/ml and his serum cortisol was [lt]2 ug/dl, requiring replacement therapy. As far as we know, this is the first reported case of such a dramatic response to combined Cisplatin and Etoposide therapy for pituitary carcinoma.[br][br]Disclosures: JWF: Investigator, Corcept Therapeutics; Consultant, Corcept Therapeutics. Nothing to Disclose: GB, HD, NM, RC, DFK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2346 202 1705 SUN-722 PO55-01 Sunday 1502 2012


1499 ENDO12L_SUN-723 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Remission of Hypercortisolism and Disappearance of an Aggressive Corticotropinoma after Temozolomide Therapy Oscar D Bruno, Maria G Pallotta Estudios Metabolicos y Endocrinos, Buenos Aires, Argentina; Hospital Italiano, Buenos Aires, Argentina [bold]Backround:[/bold] Aggressive pituitary tumors are invasive macroadenomas refractory to surgical and medical treatment, showing tendency to growth and implicating a bad vital prognosis. Treatment with the alkilating agent temozolomide (TMZ) has shown variable responses in a limited number of cases reported in the literature, with tumor volume reduction and control of the disease in many of them.[br][bold]Clinical Case:[/bold] We show the case of a 52-year old woman bearing an aggressive macrocorticotropinoma invading both cavernous sinuses, with suprasellar extension and blindness of her left eye, operated on 5 times (1 transphenoidal, 4 transcraneal) and irradiated with LINAC. Pathologic anatomy of the last surgery showed immunoreactivity for ACTH in 100% of the tumor cells. She was hypertensive, had diabetes mellitus treated with insulin, presented discrete rounding of the face, supraclavicular fat pads and complained of diminished muscle strength and easy fatigue. A 24h-urinary cortisol was 351 [micro]g/24 h (nv 20-100), ACTH 97 pg/ml (nv [lt] 46). Ketoconazol, 400 mg/day was started, and temozolomide 180 mg/d, after ondasetron 8 mg p.o., to prevent nausea and vomiting, was also indicated for 5 days monthly. Therapy was well tolerated, besides constipation. After the 4[sup]th[/sup] series of temozolomide, the patient recovered vision of the affected eye, lost 10 kg, developed hypoglycemic crisis and hypotension, so insulin and antihypertensive drugs were stopped. Ketoconazol was discontinued because of signs of steroid deprivation and subnormal urinary cortisol values at 13 [micro]g/24 h. MRI showed a 50% reduction in tumor size. Thereafter, she was treated with hydrocortisone 20-30 mg/day and kept on temozolomide. She remained asymptomatic carrying out a normal life. MRI at the 6[sup]th[/sup] month showed a nearly total disappearance of the pituitary mass. After more than 2 years, the patient remains asymptomatic and her last MRI did not show signs of adenoma.[br][bold]Conclusion: [/bold]Temozolomide induced a complete disappearance of the macrocorticotropinoma and full control of hypercortisolemia, which is an infrequent effect. It remains to be determined if the treatment must be kept, reduced to maintenance doses or stopped.[br][br]Nothing to Disclose: ODB, MGP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 839 202 1706 SUN-723 PO55-01 Sunday 1503 2012


1500 ENDO12L_SUN-724 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Temozolomide Treatment in Malignant Prolactinoma: Case Report Selcuk Dagdelen, Safak Akin, Ayse Kars, Ibrahim Ziyal, Duygu Sezen, Aysenur Cila, Figen Soylemezoglu, Tomris Erbas Hacettepe Medical School, Ankara, Turkey; Hacettepe Medical School, Ankara, Turkey; Hacettepe Medical School, Ankara, Turkey; Hacettepe Medical School, Ankara, Turkey; Hacettepe Medical School, Ankara, Turkey; Hacettepe Medical School, Ankara, Turkey We report the case of a male with a malignant prolactinoma resistant to cabergoline; he was successfully treated via CyberKnife radiosurgery and temozolomide therapy.[br]A 60-year-old man presented with headache and decreased libido for the last three years. Thereafter, pitosis in the right eye and diplopia appeared. The assessment of pituitary function documented elevated PRL levels ([gt]200 ng/ml) and a severe anterior hypopituitarism. Visual field assessment revealed bilateral temporal hemianopia. The pituitary MRI showed a giant intrasellar and suprasellar pituitary mass (35x33x50 mm) invading the third ventricle and cavernous sinuses, and compressing the optic chiasm and right temporal lobe. The patient underwent surgery through the transcranial route and a partial removal of adenoma. Immmunohistochemically, the tumor cells were positive for synaptophysin and PRL. After operation, the patient remained without follow up for several years. Three years later, on pituitary MRI, an aggressive sellar mass was noted, with leptomenengial involvement of posterior fossa. Patient underwent left suboccipital craniotomy to obtain a specimen for tumor histological diagnosis. Histological study of a biopsy sample revealed positive immunostaining for PRL with mitotic activity and high Ki-67 labeling index (9-10%). Thus a diagnosis of malignant prolactinoma with intracranial metastases was made. Laboratory tests revealed high levels of serum PRL ([gt]2000 ng/ml). Therapy was initiated with cabergoline. However, serum PRL levels remained refractory to medical intervention. Therefore, five years after the initial evaluation, the patient was started on the adjuvant therapy, with temozolomide. Fractionated CyberKnife radiosurgery was also administered. Two months later, PRL decreased gradually from 1773 ng/mL to a nadir of 275 ng/mL. The effect on tumor volume in response of therapy was evaluated four months after. A marked reduction in tumor volume was noted. In conclusion, this case-report suggests that in malignant prolactinomas, a multimodal approach is necessary with radiotherapy and temozolomide therapy.[br][br]Nothing to Disclose: SD, SA, AK, IZ, DS, AC, FS, TE 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 386 202 1707 SUN-724 PO55-01 Sunday 1504 2012


1501 ENDO12L_SUN-725 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Prevalence and Clinical Features of Patients with Pituitary Adenomas (PA) with Ki-67 Labeling Index (Ki 67) Greater Than 3%: A Retrospective Study Mariela Glerean, Debora Katz, Valeria de Miguel, Maria Victoria Arzadun, Soledad Lovazzano, Karina Miragaya, Norma Arakaki, Gustavo Sevlever, Andres Cervio, Silvia Christiansen, Antonio Carrizo, Patricia Fainstein Day Hospital Italiano, Buenos Aires, Argentina; Instituto Fleni, Buenos Aires, Argentina [bold]Introduction[/bold]: Ki-67 is an immunohistochemical marker of proliferation during G1, G2-M and S phases of cell cycle. Whereas most PA have an indolent behavior, there is a subset of aggressive or atypical PA that have been defined by the WHO one of whose pathologic features is a Ki-67 of more than 3%. Also, Ki-67 has been reported as the pathologic marker that best correlated with clinical postsurgical course of patients with aggressive prolactinomas.[br][bold]Aim[/bold]: 1- To assess the prevalence of PA with Ki-67 of more than 3%. 2- Describe the clinical features of patients that harbored these tumors.[br][bold]Subjects and Methods[/bold]: Ki-67 has been routinely determined by MIB-1 antibody and calculated by counting 100 cells. We retrospectively identified 174 patients with surgically treated PA between July 2008 and July 2011 in which Ki-67 was sistematically studied. We divided them in two groups according Ki-67: greater than 3% (Ki-67 HIGH) and 3% or less (Ki-67 LOW) and established the prevalence of KI-67 HIGH. We then compared clinical characteristics of both groups: age, gender, tumor size by MRI, tumor type according hormone secretion and treatments needed after surgery.[br][bold]Results: [/bold]We found 32 patients (18.4%) in the HIGH Ki 67 group and 142 (81.6%) in the LOW Ki 67 group. The mean age in the former was 39 [plusmn]14 years and was significantly lower than the Low Ki 67 patients (p 0.003). There was no differences in gender distribution. Most of the tumors were macroadenomas en both groups. Non functioning adenomas were the most common type in both groups. Among the HIGH KI 67 there were 14 patients (43%) who need a second or more surgical treatments, 19 patients (65.5%) who need medical treatment, 12 patients (41.3%) who required radiotherapy treatment (included radiosurgery) and all this items were significantly different higher than LOW Ki 67 group.[br][bold]Conclusions[/bold]: The prevalence of operated PA showing Ki-67 greater than 3% was 18.4%. Patients were younger, prolactinomas and non-functioning were the most frequent type and patients with this atypical tumors required reoperation and/or radiotherapy more frequently. Ki-67 greater than 3% seems a useful tool to identify potential aggressive behavior in operated PA.[br][br]Nothing to Disclose: MG, DK, VdM, MVA, SL, KM, NA, GS, AC, SC, AC, PFD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 697 202 1708 SUN-725 PO55-01 Sunday 1505 2012


1502 ENDO12L_SUN-726 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) What Causes a Prolactinoma To Be Aggressive or a Pituitary Carcinoma? Jana W Phillips, Honey E East, Sarah E French, Eugen Melcescu, Robert D Hamilton, William C Nicholas, Jonathan F Fratkin, Andrew D Parent, Gustavo Luzardo, Christian A Koch University of Mississippi Medical Center, Jackson, MS; University of Mississippi Medical Center, Jackson, MS; University of Mississippi Medical Center, Jackson, MS [bold]Introduction[/bold][br]Malignant prolactinoma is exceedingly rare and cannot be diagnosed on histological grounds. Instead, the presence of metastases within or outside the CNS is required. Treatment options are limited, and survival is poor.[br][bold]Case[/bold][br]A 25-yo WM presented to his PCP with severe headaches and sx of hypogonadism. Brain MRI showed a 4 x 3 x 2 cm pituitary tumor invading the left cavernous sinus. Labs were pertinent for elevated prolactin (470 ng/ml) and low testosterone. Transsphenoidal pituitary resection was undertaken. Pathology revealed prolactinoma with Ki-67 of 23%. Postoperatively, prolactin remained elevated (725-891 ng/ml); cabergoline was increased to 1 mg three times weekly. In spite of that, prolactin was 3507 ng/ml five months postop. Repeat MRI revealed extension of the tumor with optic chiasm compression and left orbit invasion. The pt developed acute left vision loss with ophthalmoplegia and underwent left frontotemporal craniotomy and tumor resection. Cabergoline was increased to 1 mg daily. Prolactin remained elevated (770 ng/ml), and he underwent proton beam radiation. He developed right eye blindness with MRI showing tumor in the right cavernous sinus and a dural based mass believed to be a metastasis. Considered too high risk for debulking surgery, he instead underwent gamma knife irradiation. This shrank the right cavernous sinus tumor mass, while the right temporal mass enlarged. A trial of temozolomide was unsuccessful. He died 31 months after his first surgery.[br][bold]Discussion[/bold][br]Similar to other neuroendocrine tumors (NET) such as pheochromocytoma, the dx of pituitary carcinoma is made only after discovery of metastases. Proliferation markers incl. high Ki-67/MIB-1 suggest rapid disease progression and malignant potential, while low level MGMT immunoexpression seems to be correlated with a more favorable response to temozolomide, a last resort drug. Pituitary carcinomas and NET can change their hormone expression profile which in part depends on their grade of differentiation. Prolactinomas can be invasive but rarely are malignant at first detection. It is poorly understood which factors trigger such biological behavior. Resistance to cabergoline therapy occurs rarely and is related to a decrease in D2 receptor gene transcription as well as a decrease in the G protein coupling the D2 receptor to adenylyl cyclase. Evidence for rare endocrine tumors is limited and built by individual case studies.[br][br]1.Dudziak K et al. Pituitary carcinoma with malignant growth from first presentation and fulminant clinical course-case report and review of the literature. J Clin Endocrinol Metab 2011;96:2665-2669. 2.Singer J, Koch CA, Kassahun W, et al. A patient with a large recurrent pheochromocytoma demonstrating the pitfalls of diagnosis. Nat Rev Endocrinol 2011;7(12):749-755. 3.Bush ZM et al. Temozolomide treatment for aggressive pituitary tumors: correlation of clinical outcome with O(6)-methylguanine methyltransferase (MGMT) promoter methylation and expression. J Clin Endocrinol Metab 2010;95(11):E280-90. 4.Raverot G et al. Temozolomide treatment in aggressive pituitary tumors and pituitary carcinomas: a French multicenter experience. J Clin Endocrinol Metab 2010;95:4592-4599. 5.Syro LV et al. Treatment of pituitary neoplasms with temozolomide: a review. Cancer 2011;117(3):454-462. 6.Miehle K, Tannapfel A, Lamesch P, Borte G, Schenker E, Kluge R, Ott RA, Wiechmann V, Koch M, Kassahun W, Paschke R, Koch CA. Pancreatic neuroendocrine tumor with ectopic adrenocorticotropin production upon second recurrence. J Clin Endocrinol Metab. 2004 Aug;89(8):3731-6. 7.Oh MC, Aghi MK. Dopamine agonist-resistant prolactinomas. J Neurosurg 2011;114(5):1369-79. 8.Molitch ME. The cabergoline-resistant prolactinoma patient: new challenges. J Clin Endocrinol Metab. 2008 Dec;93(12):4643-5. 9.Dillard TH, Gultekin SH, Delashaw JB Jr, Yedinak CG, Neuwelt EA, Fleseriu M. Temozolomide for corticotroph pituitary adenomas refractory to standard therapy. Pituitary. 2011 Mar;14(1):80-91.[br][br]Nothing to Disclose: JWP, HEE, SEF, EM, RDH, WCN, JFF, ADP, GL, CAK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 846 202 1709 SUN-726 PO55-01 Sunday 1506 2012


1503 ENDO12L_SUN-727 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Case of Normoprolactinemic Invasive Basal Skull Prolactinoma [mdash] Management Difficulties ChenchiReddy Kankara, Duncan L Browne Royal Cornwall Hospital, Truro, UK We report a complex case of basal skull prolactinoma presenting with CSF rhinorrhoea and without elevated prolactin. Our case is a 41 year woman who presented 2 years ago with streptococcal meningitis and CSF rhinorrhea. Initial imaging suggested right sphenoid tumor eroding the skull base involving structures in anterior and middle cranial fossa. After the tumor was biopsied by ENT team, she developed signs of recurrent meningitis, therefore referred to neurosurgery. She had partial resection of the tumor transcranially. Biopsy suggested prolactinoma. However her prolactin was only 38 mIU/L (ref. range 102-496). Glucagon test suggested normal cortisol and GH reserve. Water deprivation test was borderline for diabetes insipidus. Her periods remained normal though she has long standing history of mild galactorrhoea on palpation of breasts. Postoperatively she developed diplopia, bilateral visual field loss and optic atrophy. Despite her prolactin being normal, due to the invasiveness and associated complications of the tumor she was commenced on 500 micrograms of cabergoline on alternate days. As prolactin was not helpful in monitoring, her tumor was monitored by MRI at 6 monthly intervals, along with visual fields. There was no significant change in the residual tumor which was extending into the right cavernous sinus, sphenoid sinus, without suprasellar extension but optic chiasm was dragged inferiorly and was inseparable from the tumor. There was no change in her visual field defects. Her CSF rhinorrhea recurred last year and underwent transsphenoidal closure of the leak. Following this her right visual acuity sharply deteriorated and developed symptoms of Charles Bonnet syndrome spectrum. She is under regular follow up in the endocrine and ophthalmology clinics.[br]This case highlights the difficulties in management and follow up of a rare presentation of normoprolactinemic invasive prolactinoma.[br][br]Nothing to Disclose: CK, DLB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2246 202 1710 SUN-727 PO55-01 Sunday 1507 2012


1504 ENDO12L_SUN-728 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Rare Case of Atypical Pituitary Adenoma: An Intermediate Form of Adenoma between the Common Benign Adenoma and the Exceedingly Rare Pituitary Carcinoma Cherie Lisa Vaz, Ajay Chaudhuri, Sandeep Dhindsa State University of New York at Buffalo, Buffalo, NY Introduction We present a case of aggressive pituitary adenoma with atypical behavior treated with surgery, radiation [amp] chemotherapy.[br]Case Presentation A 46 y/o woman presented with nausea [amp] vision loss R[gt]L. She had no symptoms of amenorrhea hypothyroidism or hypoadrenalism. Brain MRI showed 4.7x3.6x3.6cm pituitary macroadenoma with mass effect on optic chiasm, internal carotid artery encasement, extension into cavernous sinus, R foramen ovale, orbital apex, sphenoid [amp] ethmoid sinus, prepontine cistern causing mass effect on midbrain [amp] pons. Pituitary hormone profile was TSH 1.41mcU/ml(0.4-5) FT4 0.53ng/ml(0.8-1.8) ACTH 20pg/ml(6-46) Prolactin 20.6ng/ml(0.5-25) FSH 1.4mU/ml(1-25) LH 0.2mU/ml(0.1-77) GH 7.9ng/ml cortisol 2.9 mcg/dl(5-25). She underwent tumor debulking [amp] optic nerve decompression. Postop MRI showed residual tumor extension into medial brain structures. Tumor pathology was sparsely granular chromophobe staining pattern, chromogrannin immunohistochemistry + CAM 5.2 [amp] cytokeratin + fibrous bodies. Ki67 labelling index 10%. Pathologic classification: nonfunctional chromophobe adenoma. Her symptoms [amp] vision improved but with residual visual loss. Postop pituitary hormone profile was TSH 0.088mcU/ml(0.4-5) FT4 0.54ng/dl(0.8-1.8) IGF 532ng/ml(288-736) ACTH 13pg.ml(5-27) am cortisol 10.6ng/ml(4-22). She received DDAVP for transient diabetes insipidus, thyroxine for secondary hypothyroidism [amp] hydrocortisone taper. 1 month follow up CT showed doubling of tumor volume, complete encasement of optic nerve with extension into orbit [amp] cranial base pathways down to hypoglossal canal. She underwent gammaknife therapy [amp] temozolomide chemotherapy.[br]Discussion This is a rare case of aggressive benign adenoma with marked progression within a month of debulking. While there are no endocrinologic, neuroradiologic [amp] histologic criteria to distinguish pituitary adenoma [amp] carcinoma, MIB-1 index of 10%, invasive growth pattern [amp] mitotic activity predicted this adenoma[apos]s aggressive biologic behavior(1). As pituitary carcinomas are thought to arise from transformation of benign invasive adenomas(2,3), given the potential for malignant transformation [amp] high risk of immediate revision surgery, temozolomide chemotherapy [amp] radiation was initiated. Case reports with temozolomide show beneficial results(4,5). Longitudinal studies on response of atypical adenomas to chemotherapy are needed. We are monitoring our patient[apos]s response to radiation [amp] chemotherapy with imaging.[br][br]1. Zada G et al J Neurosurg. 2011 Feb;114(2):336-44. 2. Scheithauer BW et al Neurosurgery2005;56:1066[ndash]1074. 3. Kaltsas GA et al J Clin Endocrinol Metab2005;90:3089[ndash]3099. 4. Colao A et al Expert Opin Pharmacother2011Jul;12(10):1561-70. 5. Losa M et al Eur J Endocrinol. 2010 Dec;163(6):843-51.[br][br]Nothing to Disclose: CLV, AC, SD 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1134 202 1711 SUN-728 PO55-01 Sunday 1508 2012


1505 ENDO12L_SUN-729 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) First Case of a Pituitary Aggressive Corticotroph Macroadenoma Concomitant with a Spindle Cell Oncocytoma Emmanuel Sonnet, Romuald Seizeur, Isabelle Quintin-Rouhe, Veronique Kerlan CHRU, Brest, France, Metropolitan; CHRU, Brest, France, Metropolitan; CHRU, Brest, France, Metropolitan Spindle cell oncytoma (SCO) is a rare non-adenomatous pituitary tumor (only 16 cases described). We report the case of a woman with this type of tumor concomitant with a corticotroph macroadenoma.[br]A 78-year-old woman presented headache, visual blurring, asthenia. Visual field assessment showed a bitemporal hemianopsia. The MRI scan revealed an extensive sellar and suprasellar lesion with compression of the visual pathways and third ventricle obstruction. Endocrinologic evaluation found a pituitary insufficiency except for ACTH. ACTH concentration was moderatly high (16 pmol/l; N[lt]12), but basal cortisol, after standard DXM test and UFC rates were normal. PRL levels were 3 times elevated. There was no diabetes insipidus. The patient underwent an endoscopic transnasal partial resection. The histological examination revealed 2 contingents: an endocrine ACTH + pattern (Ki67:3%) and another showing fascicles of spindle EMA + eosinophilic cells with numerous mitosis (Ki67:5%). This second aspect was compatible with the diagnosis of SCO. One month later, the patient had headache again, with no improvement of the visual blurring. A new MRI scan showed recurrence and an increase of the tumor. A new debulking through transcranial approach was performed. The surgeon described an aggressive tumor, with some firm and vascular parts (more peripheric) and other softer. The new histological examination confirmed the diagnosis of SCO concomitant with a corticotroph adenoma.[br]To our knowledge, this is the first case of pituitary aggressive corticotroph macroadenoma concomitant with a SCO. SCO are rare tumors, hormonally inert and can involve different organs. SCO of the pituitary is rare (only 16 cases described), first described in 2002, and recognized in the 2007 WHO classification of CNS tumors. The cells are described with an eosinophilic, granular cytoplasm and a particular immunophenotype which includes positive immunoreaction to vimentin, EMA, and S-100 and negative staining for pituitary hormones and neuroendocrine markers. First cases were described with a benign course. But other cases showed a recurrence after a first debulking. The incomplete resection was found to be a significant risk factor for recurrence, but not Ki67 rate. Surgery seems to be the only therapy, (no sensitivity to radiotherapy). In this case, the association with an aggressive corticotroph macroadenoma can suggest the use of chemotherapy such as temozolamide.[br][br]Nothing to Disclose: ES, RS, IQ-R, VK 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 445 202 1712 SUN-729 PO55-01 Sunday 1509 2012


1506 ENDO12L_SUN-730 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Recurrent Spindle Cell Oncocytoma of the Pituitary Gland Pearl Dy, Barbara Feuerstein State University of New York (SUNY) Upstate Medical University, Syracuse, NY Introduction[br]Spindle cell oncocytoma (SCO) is a rare, poorly recognized pituitary tumor, which can appear as a nonfunctioning pituitary adenoma clinically and radiologically. It is diagnosed by histopathology.[br]Case[br]We describe a case of aggressive, recurrent SCO in a 34 year old male, who complained of right lateral vision loss over 1.5 months. He had persistent headaches, cold intolerance, lack of motivation, decreased libido, occasional hot flashes and sluggishness with gradual weight gain. Opthalmologic exam confirmed right homonymous hemianopsia. Lab evaluation confirmed panhypopituitarism with FT4 0.62 ng/dL(0.90-1.70 ng/dL), TSH 1.38[micro]u/mL(0.27 -4.2 [micro]U/mL), prolactin 23.7ng/mL(4 -15.2 ng/mL), testosterone level 49ng/dL(300-1080ng/dL), FSH 1.8mU/mL(1.5-12.4 mU/mL), LH 2mU/mL(1.7-8.6 mU/mL), IGF-1 103 ng/mL(89-350 ng/mL), alpha subunit [lt]0.3 ng/mL ([lt]0.6ng/mL), cortisol 5.4 [micro]g/dL.[br]MRI documented a homogenous enhancing 3.3 cm sellar mass expanding the pituitary gland extending into the suprasellar region. Mass effect on the optic chiasm with superior displacement observed.[br]The tumor, firmer and more vascular than typical adenoma, was excised by transphenoidal resection with significant blood loss. Cytology revealed mixed spindle and epitheloid cells positive for vimentin, S100 protein, focal epithelial membrane antigen, PAS and Pas-D and diffuse nuclear expression of TTF-1; negative staining for neuroendocrine hormones and peptides.[br]Postoperatively, he required levothyroxine, hydrocortisone, testosterone gel and DDAVP. He subsequently had headaches and visual deficits, required another transphenoidal resection 5 months later, and after another 5 months, MRI showed a 2.2 cm pituitary mass, for which he underwent Gamma Knife radiosurgery.[br]Conclusion[br]As seen in this case, SCO can be difficult to dissect as they are firm, fibrous, highly vascular and adherent to surrounding structures. Although earlier reports of SCO suggest a benign clinical course, recurrences have been more recently reported.[br]Our case demonstrates that this tumor can be aggressive and difficult to treat surgically. SCO has unique immunophenotypic and ultrastructural features that facilitate its histopathological diagnosis. It should be considered in the differential diagnoses of sellar tumors. Its hypervascularity poses a challenge to complete resection and represents a significant risk factor for recurrence. While resection is the main approach, radiosurgery may be a reasonable option.[br][br]1. Roncaroli F, Scheithauer B, Cennacchi G, et al. (2002) Spindle cell oncocytoma of the adenohypophysis: a tumor of folliculostellate cells? Am J Surg Pathol 26: 1048-1055. 2. Romero-Rojas A, Melo-Uribe M, Barajas-Solano P, et al. (2011) Spindle cell oncocytoma of the adenohypophysis. Brain Tumor Pathol 28: 359-364. 3. Vajtai I, Beck J, Kappeler A, ekkehard H (2011) Spindle cell oncocytoma of the pituitary gland with follicle-like component: organotypic differentiation to support its origin from folliculo-stellate cells. 4. Kloub O, Perry A, Tu P, et al (2005) Spindle cell oncocytoma of the adenohypophysis Report of two recurrent cases. Am J Surg Pathol 29: 247-253. 5. Ogiwara H, Dubner S, Shafizadeh S , et al (2011) Spindle cell oncocytoma of the pituitary and pituitcytoma: two tumors mimicking pituitary adenoma. Surg neurol Int: 2: 116. 6. Dahiya S, Sarkar C, Hedley-White T, et al. (2005) Spindle cell oncocytoma of the adenohypophysis; report of two cases. Acta Neuropathol 110: 97-99.[br][br]Nothing to Disclose: PD, BF 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1007 202 1713 SUN-730 PO55-01 Sunday 1510 2012


1507 ENDO12L_SUN-731 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Hot Flushes in a Male [mdash] An Unusual Presentation of a Rare Pituitary Tumor Krithi B Ramesh, Joanna Miragaya, Aradhana Addepalli, Abdul Abou-Samra Detroit Medical Center/Wayne State University, Detroit, MI Introduction: Pituicytoma, a rare primary tumor of the neurohypophysis, usually presenting as pituitary adenoma. They originate from modified glial cells called pituicytes.[br]Case Description: 72 year old male was referred for evaluation of hot flushes which started 2 year ago. He also experienced decreased libido, fatigue and bilateral blurred vision but no headache, gynacomastia, weight loss, galactorrhea or symptoms of thyroid dysfunction. Physical examination was unremarkable but for mild hypertension. Given this presentation, a biochemical workup initiated for carcinoid and pheochromocytoma came back normal. His symptoms were attributed to secondary hypogonadism (AM Total Testosterone 77ng/dl, Bioavailable Testosterone 37ng/dl, confirmed twice, FSH 2.4mIU/ml, and LH 1.6mIU/ml). Panhypopituitarism was eventually diagnosed (Free T4 0.7ng/dl, TSH 1.2MicroIU/ml, IGF-1 60ng/ml). Magnetic Resonance Imaging (MRI) of pituitary showed a 1x 1.3 x 1.5 cm oval suprasellar mass appearing to displace and compress optic chiasm and optic tract. A neurosurgical referral led to a transphenoidal resection during which only a partial resection was done since the risks of pituitary stalk disruption were more serious with complete resection given the patient[apos]s age. Intraoperative pathology showed pituicytoma. Postoperatively he developed new onset bitemporal hemianopsia which was attributed to pressure on the optic chiasm from hemorrhage in the surgical bed as evidenced in post-op day 2 MRI. A final diagnosis of Pituicytoma WHO grade 1 was made based on the classical histopathology findings.[br]Repeat MRI at 3 months showed an enhancement along the infundibular stalk that extended into the optic chiasm suggestive of residual tumor. Although immediate postoperatively he did not need steroid replacement, 4 months later it was initiated due to a low AM cortisol. He was also started on testosterone and levothyroxine. At 12 months, he was overall doing well with stable MRI findings, and resolved hot flushes.[br]Conclusion: Literature review indicates only few reported cases of pituicytoma. They present as nonfunctioning pituitary tumors and definitive diagnosis comes from histologic analysis. Our patient is unique in terms of the clinical presentation with hot flushes triggering workup for other causes and symptomatic improvement soon after surgery. He was eventually diagnosed as secondary hypogonadism from the pituitary adenoma that proved to be pituicytoma on histology.[br][br]Nothing to Disclose: KBR, JM, AA, AA-S 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 831 202 1714 SUN-731 PO55-01 Sunday 1511 2012


1508 ENDO12L_SUN-732 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Acidophil Stem Cell Adenoma: Clinical, Histological and Ultrastructural Features of an Aggressive Form of Pituitary Tumor Fleur Talbot, Nemanja Stojanovic, Jonathan Pollock, Ute Pohl, Federico Roncaroli, James Ahlquist Southend Hospital, Westcliff on Sea, UK; Queen[apos]s Hospital, Romford, UK; Imperial College of Medicine, London, UK A 35 year old man presented with poor vision and bitemporal hemianopia due to a 40 mm pituitary tumour compressing the optic chiasm, with minimal para-sellar extension. Serum prolactin was marginally elevated (511 mU/L, 21.3 ng/mL); there were no clinical features of hyperprolactinaemia or growth hormone excess. He underwent trans-sphenoidal surgery, vision was fully restored and he developed ACTH deficiency. Histology showed a pituitary adenoma with unusual features; there were amphophilic and eosinophilic cells, brisk mitotic activity with a high proliferation rate (Ki67 8%), and over-expression of p53. Immunohistochemistry was weakly positive for prolactin (20%) and [alpha] subunit (15%), and negative for ACTH. Interval scan revealed rapid significant tumour regrowth over 6 months, and early repeat surgery was required. Histology was similar, with greater staining for PRL and additional scanty focal staining for GH; giant nuclei were also noted. Electron microscopy demonstrated paranuclear bodies and unusually enlarged mitochondria. These are characteristic features of an acidophilic stem cell adenoma (1). He has completed pituitary radiotherapy with no further loss of pituitary function.[br]Recent reports have demonstrated that aggressive pituitary adenomas refractory to surgery and radiotherapy may respond to temozolomide, a second generation alkylating agent. It is recommended that the level of MGMT protein expressed by the tumour be measured, as high levels are associated with poor drug susceptibility. Conversely, epigenetic silencing of the MGMT gene is associated with a better temozolomide response. We anticipate that temozolomide therapy may be required in this patient; MGMT studies are awaited.[br]We report the specific histological features of an acidophil stem cell pituitary adenoma, an aggressive tumour with early recurrence. Most pituitary adenomas behave in a benign and indolent manner; aggressive behaviour is rare, and is usually associated with silent corticotroph adenomas. This case demonstrates early recurrence of an acidophil stem cell adenoma, a tumour subtype which has unusual histological and ultrastructural features and typically shows early and rapid tumour regrowth. The malignant potential of these tumours is uncertain. The identification of patients with acidophil stem cell adenoma from the clinical and histological features described is essential to allow thorough evaluation and the opportunity of early aggressive therapy for tumour control.[br][br](1) Horvath et al. Acidophil stem cell adenoma of the human pituitary: clinicopathologic analysis of 15 cases. Cancer 1981; 47:761.[br][br]Nothing to Disclose: FT, NS, JP, UP, FR, JA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1755 202 1715 SUN-732 PO55-01 Sunday 1512 2012


1509 ENDO12L_SUN-733 POSTER SESSION: Hormonally Active [amp] Aggressive Pituitary Tumors Case Reports (1:30 PM-3:30 PM) Diffuse B-Cell Lymphoma Presenting as a Pituitary Mass Diana Barb, Francisco Pasquel, Chad Holder, Nelson M Oyesiku, Adriana G Ioachimescu Emory University School of Medicine, Atlanta, GA; Emory University School of Medicine, Atlanta, GA; Emory University School of Medicine, Atlanta, GA Lymphoma involving the pituitary is exceedingly rare (0.1-0.3 % of pituitary tumors). Metastatic involvement of the pituitary from wide spread lymphoma is also very uncommon (0.5%).[br]A 46-year-old woman presented with a 10-month history of headaches. She was otherwise healthy except for irregular menses since age 31, and amenorrhea since 44. She did not have cushingoid or acromegalic appearance and visual fields were normal. MRI revealed a 2 cm sellar and suprasellar mass with invasion of the clivus, bilateral cavernous sinuses and right parasellar region. She had normal CBC and chemistry panel except an elevated alkaline phosphatase level twice above normal. Hormonal testing showed normal thyroid and adrenal axis, elevated FSH and LH in the menopausal range and a mild elevation of IGF-1 of 358 ng/mL (repeated 338, nl: 54-307 ng/ml). GH was normal (0.13 ng/mL) but showed a paradoxical increase during OGTT at 60 and 90 minutes (0.15 ng/mL at 30 minutes, 4.3 ng/mL at 60, 5.2 ng/mL at 90 and 0.64 ng/mL at 120 minutes). Baseline glucose was 88 and increased to 111 mg/dL at 120-minutes. The patient underwent transsphenoidal surgery. Intraoperative biopsy of the tumor showed diffuse large B-cell lymphoma (DLBCL), strongly positive for CD20, CD79a, and PAX5. Further investigation with PET-CT and spine MRI showed metastatic involvement of the thoracic spine at T7 and 7th rib and abnormal bone marrow within the skull base and right frontal skull. However, bone marrow biopsy and CSF cytology did not show lymphoma involvement. The patient started chemotherapy with R-HCVAD and intrathechal therapy as appropriate for diffuse large cell B lymphoma stage IV.[br]We present a rare case of pituitary involvement by lymphoma with mild elevation of IGF-1 level without clinical signs of GH excess. Lymphoma should be considered in the differential diagnosis of a sellar mass with invasive character. Epidemiologic studies showed that high IGF-I levels are associated with increased risk of cancers. In addition, in vitro mRNA expression of the B cell lymphoma tumor repressor gene Bcl-6 was shown to be decreased by GH in vitro (1). In humans, Bcl-6 translocation or mutation was associated with DLBCL (2). Our case opens translational research avenues regarding the possible role of excess GH in the pathogenesis of DLBCL.[br][br](1) Chen Y et al., Endocrinology 2009;150(8):3645-54. (2) Schneider et al., Seminars in Diagnostic Pathology 2011;28:167-177.[br][br]Nothing to Disclose: DB, FP, CH, NMO, AGI 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 2233 202 1716 SUN-733 PO55-01 Sunday 1513 2012


1510 ENDO12L_SUN-734 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Long-Term Pasireotide Use Leads to Significant and Sustained Improvements in the Signs and Symptoms of Cushing Disease: 24-Month Results from a Randomized Phase III Study Jerome Bertherat, Jochen Schopohl, William H Ludlam, Mario Maldonado, Andrew Trovato, Gareth Hughes, Feng Gu, Luiz R Salgado, Rosario Pivonello H[ocirc]pital Cochin, Paris, France; University of Munich, Munich, Germany; Swedish Neuroscience Institute, Seattle, WA; Novartis Pharma AG, Basel, Switzerland; Novartis Pharmaceuticals Corporation, East Hanover, NJ; Peking Union Medical College Hospital, Beijing, China; University of S[atilde]o Paulo Medical School, S[atilde]o Paulo, Brazil; University of Naples [apos]Federico II[apos], Naples, Italy [bold]Introduction:[/bold] Cushing[apos]s disease results in increased mortality and morbidity. A large, randomized, Phase III study of pasireotide in patients with Cushing[apos]s disease (n=162) showed pasireotide treatment provided a rapid reduction in UFC and significant improvements in signs and symptoms. Here, we report the results of a 12-month extension to this trial.[br][bold]Methods:[/bold] Patients with persistent, recurrent or [italic]de novo[/italic] (if not surgical candidates) Cushing[apos]s disease and UFC levels [ge]1.5xULN were randomized to pasireotide 600[mu]g (n=82) or 900[mu]g (n=80) sc bid. Patients who had mean UFC[le]ULN at 12m or who were achieving clinical benefit from pasireotide were eligible to enter an extension phase. Dose titration was permitted at the investigator[apos]s discretion (min: 300[micro]g sc bid; max: 1200[micro]g sc bid). A last observation carried forward analysis was used for UFC, ACTH and cortisol. Signs and symptoms (observational analysis) were recorded every 3m during the extension as secondary endpoints.[br][bold]Results:[/bold] Fifty-eight patients entered the extension. The mean [95%CI] change in UFC from baseline was [minus]54.7% [[ndash]71.8, [ndash]37.6] at 12m and [minus]59.5% [[ndash]68.6, [ndash]50.5] at 24m. Similarly, plasma ACTH and serum cortisol decreased from baseline. An improvement in the signs and symptoms of Cushing[apos]s disease was observed over 24m. The mean change from baseline in SBP was [minus]8.4mmHg at 12m [[minus]12.3, [minus]4.5] and [minus]11.3mmHg [[minus]15.0, [minus]7.5] at 24m. The mean change from baseline in DBP was [minus]4.0mmHg at 12m [[minus]6.9, [minus]1.2] and [minus]7.2mmHg at 24m [[minus]10.4, [minus]3.9]. At 12m, the mean change from baseline in BMI was [minus]9.1% [[minus]10.8, [minus]7.4] and this was maintained during the extension phase, with a mean change of [minus]9.6% [[minus]12.1, [minus]7.2] reported at 24m. The significant decrease in BMI resulted from a significant decrease in patients[apos] weight at 12m and 24m ([minus]9.1% [[ndash]10.8, [ndash]7.4] and [minus]9.6% [[ndash]12.1, [ndash]7.2], respectively). The mean change from baseline in total cholesterol levels was [minus]8.5% [[minus]13.4, [minus]3.6] and [minus]5.8% [[minus]11.4, [minus]0.1] at 12m and 24m, respectively. 24m safety results are reported in a separate abstract.[br][bold]Conclusion:[/bold] Long-term pasireotide treatment results in sustained reductions in UFC, plasma ATCH and serum cortisol. Significant improvements in blood pressure, BMI, weight and cholesterol were also sustained to 24m, supporting the use of pasireotide as a treatment for patients with Cushing[apos]s disease.[br][br]Sources of Research Support: Novartis Pharmaceuticals.[br][br]Disclosures: JB: Medical Advisory Board Member, Novartis Pharmaceuticals. WHL: Speaker, Pfizer, Inc., Novartis Pharmaceuticals, Ipsen; Medical Advisory Board Member, Corcept Therapeutics, Novartis Pharmaceuticals, Ipsen, Endo Pharmaceuticals Inc. MM: Employee, Novartis Pharmaceuticals. AT: Employee, Novartis Pharmaceuticals. GH: Employee, Novartis Pharmaceuticals. RP: Consultant, Novartis Pharmaceuticals. Nothing to Disclose: JS, FG, LRS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1720 203 1717 SUN-734 PO22-02 Sunday 1514 2012


1511 ENDO12L_SUN-735 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Pasireotide as a Long-Term Treatment for Patients with Cushing Disease: 24-Month Safety Results from a Randomized Phase III Study Jerome Bertherat, William H Ludlam, Rosario Pivonello, Mario Maldonado, Andrew Trovato, Gareth Hughes, Feng Gu, Jochen Schopohl, Luiz R Salgado H[ocirc]pital Cochin, Paris, France; Swedish Neuroscience Institute, Seattle, WA; University of Naples [apos]Federico II[apos], Naples, Italy; Novartis Pharma AG, Basel, Switzerland; Novartis Pharmaceuticals Corporation, East Hanover, NJ; Peking Union Medical College Hospital, Beijing, China; University of Munich, Munich, Germany; University of S[atilde]o Paulo Medical School, S[atilde]o Paulo, Brazil [bold]Introduction:[/bold] A large, randomized, 12-month, Phase III study of pasireotide in patients with Cushing[apos]s disease showed rapid and sustained decreases in UFC levels and significant improvements in signs and symptoms. The safety profile of pasireotide was similar to that of other somatostatin analogues, with the exception of hyperglycemia-related AEs (reported in 72.8% of patients, including hyperglycemia [40.1%] and diabetes mellitus [27.2%]). Here, we report 24 months[apos] safety data.[br][bold]Methods:[/bold] Patients with persistent, recurrent or [italic]de novo[/italic] (if not surgical candidates) Cushing[apos]s disease and UFC levels [ge]1.5xULN were randomized to receive pasireotide 600[mu]g (n=82) or 900[mu]g (n=80) sc bid. Those with mean UFC[le]ULN or who achieved clinical benefit from pasireotide at month 12 (end of core) were eligible to enter the extension. Dose titration was permitted at the investigator[apos]s discretion (min: 300[micro]g sc bid; max: 1200[micro]g sc bid). Data on AEs were collected over 24 months.[br][bold]Results:[/bold] Fifty-eight patients entered the extension phase and 39 completed 24 months[apos] treatment. The mean change in UFC levels from baseline was [minus]59.5% (95% CI: [ndash]68.6, [ndash]50.5) at month 24 (last observation carried forward). Most patients (98.1%) experienced [ge]1 AE over 24 months, mainly GI disorders of mild/moderate severity. There were no deaths. Over 24 months, the most common AEs (all grades) were diarrhea (58.6% patients), nausea (52.5%), hyperglycemia (40.1%), cholelithiasis (32.1%) and diabetes mellitus (29.6%). Of patients who experienced AEs over the 24 months, diabetes mellitus was reported in only 4 new patients (6.9%) during the extension, mild-to-moderate cholelithiasis in 3 (5.2%), a [gt]30ms increase in QTcF in 2 (3.5%), newly occurring QTcF [gt]480ms in 1 (1.7%) and hyperglycemia in no new patients. Mean HbA[sub]1c[/sub] increased from 5.8% at baseline to 7.2%, 6.9% and 6.8% at months 12, 18 and 24, respectively. Over 24 months, [ge]1 protocol-defined SAE was experienced by 25.9% of patients, although only 2 patients experienced a SAE during the extension. Three patients discontinued because of an AE during the extension. 24-month efficacy results are reported in a separate abstract.[br][bold]Conclusion:[/bold] The 24-month safety profile of pasireotide was similar to that reported at 12 months. Hyperglycemia AEs were reported in no new patients during the extension. These results support the use of pasireotide as a long-term treatment for Cushing[apos]s disease.[br][br]Sources of Research Support: Novartis Pharmaceuticals.[br][br]Disclosures: JB: Medical Advisory Board Member, Novartis Pharmaceuticals. WHL: Speaker, Pfizer, Inc., Novartis Pharmaceuticals, Ipsen; Medical Advisory Board Member, Corcept Therapeutics, Novartis Pharmaceuticals, Ipsen, Endo Pharmaceuticals Inc. RP: Consultant, Novartis Pharmaceuticals. MM: Employee, Novartis Pharmaceuticals. AT: Employee, Novartis Pharmaceuticals. GH: Employee, Novartis Pharmaceuticals. Nothing to Disclose: FG, JS, LRS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1734 203 1718 SUN-735 PO22-02 Sunday 1515 2012


1512 ENDO12L_SUN-736 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Pasireotide Treatment Is Associated with Clinically Meaningful Improvements in Health-Related Quality of Life in Cushing Disease: Results from a Large, Randomized, Double-Blind, Phase III Trial Monica Gadelha, Xavier Badia, Wojciech Zgliczynski, Lesly Portocarrero-Ortiz, Mario Maldonado, Andrew Trovato, Anna Forsythe, Lauren Nelson, Lori McLeod, Christophe De Block, Susan M Webb Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; IMS Health, Barcelona, Spain; Centre for Postgraduate Medical Education, Warsaw, Poland; National Institute of Neurology and Neurosurgery [apos]Manuel Velasco Su[aacute]rez[apos], Mexico City, Mexico; Novartis Pharma AG, Basel, Switzerland; Novartis Pharmaceuticals Corporation, East Hanover, NJ; RTI Health Solutions, Research Triangle Park, NC; University Hospital, Antwerp, Belgium; Universitat Aut[ograve]noma de Barcelona, Barcelona, Spain [bold]Introduction: [/bold]Patients with Cushing[apos]s disease (CD) have significantly impaired health-related quality of life (HRQoL). Treatment options beyond surgery are limited, which limits the ability to improve HRQoL. There are currently no approved medical therapies for CD. A randomized, Phase III study has shown that pasireotide leads to rapid and sustained decreases in urinary free cortisol (UFC). Here, we report the effects of pasireotide on patient HRQoL during this trial.[br][bold]Methods: [/bold]Patients with persistent, recurrent or [italic]de novo[/italic] (if not surgical candidates) CD and UFC levels [ge]1.5 times the upper limit of normal (ULN) were randomized to receive pasireotide 600 [mu]g (n=82) or 900 [mu]g (n=80) sc bid. Dose titration (max: 1200 [micro]g bid) was permitted after month 3. UFC control was defined as UFC[le]ULN, partial control as UFC[gt]ULN and [ge]50% reduction from baseline, and uncontrolled UFC as UFC[gt]ULN and [lt]50% reduction from baseline. HRQoL was assessed at baseline and months 3, 6 and 12 using the 12-item CushingQoL questionnaire. CushingQoL items relate to the effects of hypercortisolism and include physical appearance, anxiety and depression. Items were scored on a 5-point scale, resulting in a score of 12 (worst) to 60 (best) that was standardized to 1[ndash]100. A change in CushingQoL score of [gt]10.1 was considered to be clinically meaningful.[br][bold]Results: [/bold]Overall, HRQoL improved rapidly from 41.1 at baseline (n=159) to 49.4, 50.4 and 52.5 at months 3 (n=133), 6 (n=112) and 12 (n=76), respectively (mean increase at month 12: 11.1; 95% CI: 6.8, 15.5). There was a clinically meaningful improvement in HRQoL at month 12 in patients whose UFC levels were controlled (n=29; mean improvement: 12.8; 95% CI: 7.1, 18.5) or partially controlled at month 6 (n=17; mean improvement: 10.7; 95% CI: 0.8, 20.5). However, improvement in HRQoL in the uncontrolled group (n=30; mean improvement: 9.9; 95% CI: 2.3, 17.6) did not reach the 10.1 change threshold. The greatest HRQoL improvements ([ge]20 points) were seen in patients with the greatest UFC decreases (from [gt]10xULN to [le]5xULN; n=5). Significant correlations ([italic]P[/italic][lt]0.01) were seen between changes in CushingQoL scores and changes in mean UFC (r= [ndash]0.40; n=68), BMI (r=0.31; n=74), weight (r=0.32; n=74), and Beck depression inventory score (r= [ndash]0.59, n=72).[br][bold]Conclusion: [/bold]Pasireotide treatment significantly reduces UFC and this is correlated with an improvement in HRQoL. Clinically meaningful improvements in HRQoL were seen regardless of achieving UFC normalization.[br][br]Sources of Research Support: Novartis Pharmaceuticals.[br][br]Disclosures: MG: Principal Investigator, Novartis Pharmaceuticals. WZ: Speaker, Novartis Pharmaceuticals, Ipsen; Investigator, Novartis Pharmaceuticals, Ipsen. MM: Employee, Novartis Pharmaceuticals. AT: Employee, Novartis Pharmaceuticals. AF: Employee, Novartis Pharmaceuticals. LN: Consultant, Novartis Pharmaceuticals; Employee, RTI Health Solutions. LM: Consultant, Novartis Pharmaceuticals; Employee, RTI Health Solutions. Nothing to Disclose: XB, LP-O, CDB, SMW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1735 203 1719 SUN-736 PO22-02 Sunday 1516 2012


1513 ENDO12L_SUN-737 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Pasireotide Treatment in Cushing Disease Is Associated with Significant Improvements in Hypertension: 12-Month Results from a Large Phase III Study Rosario Pivonello, Stephan Petersenn, John Newell-Price, James W Findling, Feng Gu, Mario Maldonado, Andrew Trovato, Gareth Hughes, Luiz R Salgado, Andre Lacroix, Jochen Schopohl, Beverly MK Biller University of Naples [apos]Federico II[apos], Naples, Italy; ENDOC Center for Endocrine Tumors, Hamburg, Germany; University of Sheffield, Sheffield, UK; Medical College of Wisconsin, Milwaukee, WI; Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing, China; Novartis Pharma AG, Basel, Switzerland; Novartis Pharmaceuticals Corporation, East Hanover, NJ; University of S[atilde]o Paulo Medical School, S[atilde]o Paulo, Brazil; Centre Hospitalier de l[apos]Universit[eacute] de Montr[eacute]al, Montr[eacute]al, Canada; University of Munich, Munich, Germany; Massachusetts General Hospital, Boston, MA [bold]Introduction: [/bold]Patients with Cushing[apos]s disease have an increased risk of hypertension. In a recent Phase III trial in Cushing[apos]s disease, pasireotide rapidly reduced UFC levels and significantly improved signs and symptoms. The effects of pasireotide on hypertension were investigated as secondary objectives in this trial.[br][bold]Methods:[/bold] Patients with persistent/recurrent or [italic]de novo[/italic] (if not surgical candidates) Cushing[apos]s disease and UFC [ge]1.5 times the upper limit of normal (ULN) were randomized to pasireotide 600[mu]g (n=82) or 900[mu]g (n=80) sc bid. Systolic (SBP) and diastolic blood pressure (DBP) were evaluated by single measurements at baseline (n=162) and month 12 (n=78). Patients who had at least one of the following baseline criteria were considered to have baseline hypertension: history of antihypertensive medications; medical history of hypertension; SBP[gt]130mmHg; DBP[gt]90mmHg. Additional medications for hypertension were allowed per investigator discretion.[br][bold]Results: [/bold]At baseline, 78% of patients had hypertension (n=126). During the study, 63% (n=102) of patients took antihypertensives, 95% of whom had hypertension at baseline. Mean SBP decreased from 133.5mmHg at baseline to 126.1mmHg at month 12 ([ndash]6.1; 95% CI: [ndash]9.8, [ndash]2.4). Mean DBP decreased from 86.3mmHg to 82.8mmHg ([ndash]3.7; 95% CI: [ndash]6.2, [ndash]1.2). When stratified by UFC response, improvements were significant in those with UFC[le]ULN at month 6. Significant improvements in SBP and DBP from baseline to month 12 were seen in patients with baseline hypertension (SBP: [ndash]8.0 [[ndash]12.4, [ndash]3.6]; DBP: [ndash]4.7 [[ndash]7.7, [ndash]1.7]). No significant changes were observed in patients without baseline hypertension (SBP: 0.2 [[ndash]6.1, 6.4]; DBP: [ndash]0.4 [[ndash]4.6, 3.9]). Greater decreases in SBP and DBP were observed in patients with higher baseline SBP and DBP. A [gt]5mmHg decrease in DBP was seen in 42% (33/78) of patients at month 12. This was higher in those with baseline hypertension (50% [30/60]) than in those without (17% [3/18]). In patients with baseline hypertension, a [gt]5mmHg decrease in DBP was observed in 46% (20/44) and 63% (10/16) of those who did and did not take antihypertensives, respectively. Adverse events (AEs) of hypotension and adrenal insufficiency were reported by 11 and 9 patients, respectively; other AEs were consistent with those reported for somatostatin analogues, except the higher degree of hyperglycemia.[br][bold]Conclusions: [/bold]Pasireotide treatment was associated with significant decreases in SBP and DBP in patients with Cushing[apos]s disease.[br][br]Sources of Research Support: Novartis Pharmaceuticals.[br][br]Disclosures: RP: Consultant, Novartis Pharmaceuticals. SP: Speaker, Novartis Pharmaceuticals, Ipsen, Pfizer, Inc.; Medical Advisory Board Member, Novartis Pharmaceuticals, Ipsen, Pfizer, Inc. JN-P: Investigator, Novartis Pharmaceuticals; Speaker, Novartis Pharmaceuticals. JWF: Investigator, Corcept Therapeutics, Novartis Pharmaceuticals; Consultant, Corcept Therapeutics, Novartis Nutrition, Inc. MM: Employee, Novartis Pharmaceuticals. AT: Employee, Novartis Pharmaceuticals. GH: Employee, Novartis Pharmaceuticals. AL: Investigator, Novartis Pharmaceuticals; Speaker, Novartis Pharmaceuticals. BMKB: Consultant, Novartis Pharmaceuticals, Corcept Therapeutics. Nothing to Disclose: FG, LRS, JS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1715 203 1720 SUN-737 PO22-02 Sunday 1517 2012


1514 ENDO12L_SUN-738 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Pasireotide LAR and Octreotide LAR Maintain Biochemical Control in Patients with Acromegaly: Extension Phase of a 12-Month Randomized, Double-Blind, Multicenter, Phase III Study Michael Sheppard, Monica Gadelha, Marcello D Bronstein, Pamela Freda, Feng Gu, Chiung-Chyi Shen, Maria Fleseriu, Karina Hermosillo Resendiz, Matthieu Ruffin, Kobby Asubonteng, Annamaria Colao University of Birmingham, Edgbaston, Birmingham, UK; Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; University of S[atilde]o Paulo Medical School, S[atilde]o Paulo, Brazil; Columbia University College of Physicians [amp] Surgeons, New York, NY; Peking Union Medical College Hospital, Beijing, China; National Yang-Ming University, Taipei, Taiwan; Oregon Health [amp] Science University, Portland, OR; Novartis Pharmaceuticals Corporation, Florham Park, NJ; Novartis Pharma AG, Basel, Switzerland; University of Naples [apos]Federico II[apos], Naples, Italy [bold]Introduction:[/bold] In a large, randomized, double-blind, Phase III trial of patients with acromegaly (n=358), pasireotide LAR was superior to octreotide LAR at providing GH[lt]2.5[mu]g/L [italic]and[/italic] normalized IGF-1 after 12m (core). The extension phase allowed patients who derived benefit during the core study to continue receiving medical therapy with their randomized treatment.[br][bold]Methods:[/bold] Patients with acromegaly (GH[gt]5[micro]g/L or GH nadir [ge]1[micro]g/L post-OGTT, and IGF-1[gt]ULN) who were [italic]de novo[/italic] with a visible adenoma on MRI or medically na[iuml]ve (defined as no previous medical therapy but could have undergone prior pituitary surgery) and completed 12m therapy with pasireotide LAR 40[ndash]60mg/28d or octreotide LAR 20[ndash]30mg/28d could enter an extension phase. Patients could continue on their randomized therapy at the investigators[apos] discretion based on GH and IGF-1 levels. Efficacy findings after 6m treatment in the extension study, and adverse events (AEs) experienced at any time during both the core study and the first 6m of the extension (during 19m of therapy), are reported here. All hyperglycemia-related AE terms were grouped.[br][bold]Results:[/bold] From the 358 patients who entered the core study, 120 patients enrolled in the extension to continue receiving pasireotide LAR (n=74) or octreotide LAR (n=46). At month 19, 45.9% (34/74) and 45.7% (21/46) had GH[lt]2.5 [micro]g/L [italic]and[/italic] normal IGF-1 in the pasireotide LAR and octreotide LAR groups, respectively; 73.0% (54/74) and 71.7% (33/46) had GH[lt]2.5 [micro]g/L, and 50.0% (37/74) and 52.2% (24/46) had normal IGF-1. The most common AEs during 19m therapy with pasireotide LAR vs octreotide LAR were diarrhea (39.9 vs 45.0%), cholelithiasis (29.8 vs 39.4%) and headache (21.9 vs 27.2%). Most AEs were mild or moderate. Hyperglycemia-related AEs were seen in 62.9% and 25.0% of pasireotide LAR and octreotide LAR patients, respectively; 6 and 3 patients discontinued due to hyperglycemia-related AEs during 19m therapy. Glucose and HbA[sub]1c[/sub] levels increased in the first 3m after initiating pasireotide therapy and remained stable to 19m.[br][bold]Conclusions:[/bold] Pasireotide LAR demonstrated superiority to octreotide LAR in providing full biochemical control (GH[lt]2.5[mu]g/L [italic]and[/italic] normal IGF-1) after 12m therapy; both pasireotide LAR and octreotide LAR patients maintained biochemical control up to 19m. Safety findings were similar to those seen in the core study; glucose and HbA[sub]1c[/sub] levels remained stable. These data suggest that pasireotide LAR may be a new treatment option for patients with acromegaly.[br][br]Sources of Research Support: Novartis Pharmaceuticals.[br][br]Disclosures: MS: Speaker, Novartis Pharmaceuticals. MG: Principal Investigator, Novartis Pharmaceuticals. MDB: Consultant, Novartis Pharmaceuticals; Investigator, Novartis Pharmaceuticals. PF: Advisory Group Member, Novartis Pharmaceuticals; Clinical Researcher, Novartis Pharmaceuticals. MF: Researcher, Novartis Pharmaceuticals, Ipsen; Medical Advisory Board Member, Novartis Pharmaceuticals; Speaker, Ipsen, Chiasma. KHR: Employee, Novartis Pharmaceuticals. MR: Employee, Novartis Pharmaceuticals. KA: Employee, Novartis Pharmaceuticals. AC: Investigator, Novartis Pharmaceuticals. Nothing to Disclose: FG, C-CS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1896 203 1721 SUN-738 PO22-02 Sunday 1518 2012


1515 ENDO12L_SUN-739 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Pasireotide LAR Provides Greater Response Than Octreotide LAR in Patients with Acromegaly Irrespective of Baseline Tumor Volume: Post-Hoc Analysis from a Large, Randomized, Double-Blind, Phase III Study Antoine Tabarin, Ariel Barkan, Sung-Woon Kim, Andrew Farrall, Karina Hermosillo Resendiz, Matthieu Ruffin, Kobby Asubonteng, Laura De Marinis CHU of Bordeaux, Bordeaux, France; University of Michigan, Ann Arbor, MI; Kyung Hee University, Seoul, Korea; University of Edinburgh, Western General Hospital, Edinburgh, UK; Novartis Pharmaceuticals Corporation, Florham Park, NJ; Novartis Pharma AG, Basel, Switzerland; Universit[agrave] Cattolica del Sacro Cuore, Rome, Italy [bold]Introduction:[/bold] Although response rates of up to 70% have been reported with currently available somatostatin analogues, when response is evaluated using both GH[lt]2.5[mu]g/L [italic]and[/italic] normalized IGF-1, 20[ndash]25% of medically na[iuml]ve patients respond at 12m. In a large, randomized, double-blind, Phase III trial, pasireotide LAR showed superiority to octreotide LAR in providing GH[lt]2.5[mu]g/L [italic]and[/italic] normal IGF-1 after 12m. Reported here are results of a post-hoc analysis evaluating the relationship between biochemical response and baseline tumor volume.[br][bold]Methods:[/bold] Patients with acromegaly (GH[gt]5[micro]g/L or GH nadir [ge]1[micro]g/L post-OGTT, and IGF-1[gt]ULN) who were [italic]de novo[/italic] with a visible adenoma on MRI or medically na[iuml]ve (no previous medical therapy but prior pituitary surgery) received pasireotide LAR 40mg/28d (n=176) or octreotide LAR 20mg/28d (n=182) for 12m. Randomization was stratified by post-surgical or [italic]de novo[/italic] status. Only patients with a 12m biochemical efficacy evaluation were included (n=134 and 151, respectively, in the pasireotide LAR and octreotide LAR arms). Responders had GH[lt]2.5[mu]g/L [italic]and[/italic] IGF-1[le]ULN at 12m. Baseline tumor volume was divided into quartiles. A macroadenoma was defined as having a tumor volume [ge]523mm[sup]3[/sup].[br][bold]Results:[/bold] Pasireotide LAR provided higher response rates than octreotide LAR in both post-surgical (56.9% [29/51] and 29.0% [18/62], respectively) and [italic]de novo[/italic] patients (34.9% [29/83] and 19.1% [17/89]). At baseline, 127 and 139 patients had evaluable tumor volume in the pasireotide LAR and octreotide LAR arms, of whom 83 and 89 were [italic]de novo[/italic]. In patients with baseline tumor volume in the lowest quartile (all microadenomas), 70% (23/33) and 26% (9/35) of patients were responders in the pasireotide LAR and octreotide LAR arms; 61% (11/18) and 27% (4/15) of the [italic]de novo[/italic] patients were responders. In patients with tumors in the 2[sup]nd[/sup], 3[sup]rd[/sup] and 4[sup]th[/sup] quartiles, 36% (12/33), 42% (13/31) and 23% (7/30) of pasireotide LAR patients and 22% (8/37), 24% (8/34) and 21% (7/33) of octreotide LAR patients were responders. Patients with tumors in the 2[sup]nd[/sup] quartile had either micro- or macroadenomas, whereas all tumors in the 3[sup]rd[/sup] and 4[sup]th[/sup] quartiles were macroadenomas.[br][bold]Conclusions:[/bold] Pasireotide LAR was superior to octreotide LAR in providing control of GH [italic]and[/italic] IGF-1 after 12m therapy. Pasireotide LAR provided benefit irrespective of baseline tumor volume.[br][br]Sources of Research Support: Novartis Pharmaceuticals.[br][br]Disclosures: AT: Consultant, Novartis Pharmaceuticals. AF: Consultant, Novartis Pharmaceuticals, Perceptive Informatics. KHR: Employee, Novartis Pharmaceuticals. MR: Employee, Novartis Pharmaceuticals. KA: Employee, Novartis Pharmaceuticals. Nothing to Disclose: AB, S-WK, LDM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1929 203 1722 SUN-739 PO22-02 Sunday 1519 2012


1516 ENDO12L_SUN-740 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Pasireotide LAR Improves Health-Related Quality of Life and Signs and Symptoms in Patients with Acromegaly: Results of a 12-Month Randomized, Double-Blind, Multicenter, Phase III Study Luciana Naves, Omar Serri, Francesco Minuto, V Pronin, Alberto Chervin, William H Ludlam, Karina Hermosillo Resendiz, Matthieu Ruffin, Kobby Asubonteng, Susan Webb University of Bras[iacute]lia, Bras[iacute]lia, Brazil; University of Montr[eacute]al, Montr[eacute]al, Canada; University of Genova, Genova, Italy; Moscow Medical Academy, Moscow, Russian Federation; Hospital Santa Lucia, Buenos Aires, Argentina; Swedish Neuroscience Institute, Seattle, WA; Novartis Pharma AG, Basel, Switzerland; Novartis Pharmaceuticals Corporation, Florham Park, NJ; Universitat Aut[ograve]noma de Barcelona and CIBERER 747, Barcelona, Spain [bold]Introduction: [/bold]Patients with acromegaly have a high symptom burden and impaired health-related quality of life (HRQoL). In a large, randomized, double-blind, Phase III trial (n=358), pasireotide LAR demonstrated superiority to octreotide LAR in achieving both GH[lt]2.5[mu]g/L [italic]and[/italic] normal IGF-1 at 12m. The effect of pasireotide LAR and octreotide LAR on HRQoL and the symptoms of acromegaly was also evaluated as part of the Phase III study, and those results are reported here.[br][bold]Methods: [/bold]Patients with acromegaly (GH[gt]5[micro]g/L or GH nadir [ge]1[micro]g/L post-OGTT, and IGF-1[gt]ULN) who were [italic]de novo[/italic] with a visible adenoma on MRI or medically na[iuml]ve (no previous medical therapy but prior pituitary surgery) received pasireotide LAR 40mg/28d (n=176) or octreotide LAR 20mg/28d (n=182) for 12 months. Dose titration to pasireotide LAR 60mg or octreotide LAR 30mg for suboptimal biochemical response was permitted, but not mandatory, at months 3 and 7, and the primary endpoint was evaluated at month 12. Changes in HRQoL and the symptoms of acromegaly were evaluated to month 12. HRQoL was assessed using the AcroQoL questionnaire, a 22-item instrument that results in scores ranging from 0 (worst HRQoL) to 100 (best HRQoL). The severity of five symptoms of acromegaly (headache, fatigue, perspiration, paresthesia and osteoarthralgia) was scored from 0 (no symptom) to 4 (very severe).[br][bold]Results: [/bold]AcroQoL scores were 73.4 (SE: 17.6) and 71 (SE: 17.42) at baseline in the pasireotide LAR and octreotide LAR arms, respectively, and improved at month 12 by 6.2 (SE: 12.8) and 4.3 (SE: 13.6) as GH and IGF-1 levels decreased. Severity of individual symptoms at baseline was similar for both treatment arms with symptom scores between 0.7 and 1.4. Improvements in all five symptoms were seen by month 12. In both treatment arms (pasireotide LAR and octreotide LAR), the largest improvement was in perspiration ([ndash]0.6 [SD: 1.14] and [ndash]0.8 [SD: 1.20]). Similar improvements were seen in fatigue ([ndash]0.4 [SD: 1.18] and [ndash]0.6 [SD: 0.96]), osteoarthralgia ([ndash]0.4 [SD: 1.07] and [ndash]0.6 [SD: 1.20]), paresthesia ([ndash]0.4; [SD: 0.90] and [ndash]0.4 [SD: 1.11]) and headache ([ndash]0.3 [SD: 1.17] and [ndash]0.4 [SD: 0.94]).[br][bold]Conclusion: [/bold]Pasireotide LAR and octreotide LAR provided improvements in symptom control and HRQoL as GH and IGF-1 levels decreased. These data support pasireotide LAR as a potential new treatment option for patients with acromegaly.[br][br]Sources of Research Support: Novartis Pharmaceuticals.[br][br]Disclosures: OS: Investigator, Novartis Pharmaceuticals. WHL: Speaker, Pfizer, Inc., Novartis Pharmaceuticals, Ipsen; Medical Advisory Board Member, Corcept Therapeutics, Novartis Pharmaceuticals, Ipsen, Endo Pharmaceuticals Inc. KHR: Employee, Novartis Pharmaceuticals. MR: Employee, Novartis Pharmaceuticals. KA: Employee, Novartis Pharmaceuticals. Nothing to Disclose: LN, FM, VP, AC, SW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1714 203 1723 SUN-740 PO22-02 Sunday 1520 2012


1517 ENDO12L_SUN-741 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Switching Patients with Acromegaly from Octreotide LAR to Pasireotide LAR Improves Biochemical Control: Crossover Extension to a Randomized, Double-Blind, Multicenter, Phase III Study Maria Fleseriu, Michael Sheppard, Marcello D Bronstein, Pamela Freda, Feng Gu, Chiung-Chyi Shen, Monica Gadelha, Karina Hermosillo Resendiz, Matthieu Ruffin, Yinmiao Chen, Annamaria Colao Oregon Health [amp] Science University, Portland, OR; University of Birmingham, Edgbaston, Birmingham, UK; University of S[atilde]o Paulo Medical School, S[atilde]o Paulo, Brazil; Columbia University College of Physicians [amp] Surgeons, New York, NY; Peking Union Medical College Hospital, Beijing, China; National Yang-Ming University, Taipei, Taiwan; Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; Novartis Pharmaceuticals Corporation, Florham Park, NJ; Novartis Pharma AG, Basel, Switzerland; University of Naples [apos]Federico II[apos], Naples, Italy [bold]Introduction:[/bold] In a large, randomized, double-blind, Phase III trial, PAS LAR was significantly more effective than OCT LAR at providing GH[lt]2.5[mu]g/L [italic]and[/italic] normal IGF-1 at 12m. Patients without full biochemical control at 12m could switch treatments.[br][bold]Methods:[/bold] Medically na[iuml]ve patients with active acromegaly (GH[gt]5[micro]g/L or GH nadir [ge]1[micro]g/L post-OGTT, and IGF-1[gt]ULN) who were [italic]de novo[/italic] with a visible adenoma on MRI or post-surgical, and who completed 12m therapy with PAS LAR 40[ndash]60mg/28d or OCT LAR 20[ndash]30mg/28d, could enter an extension phase. Patients with GH[lt]2.5[mu]g/L [italic]and[/italic] normal IGF-1 at 12m could continue on their randomized therapy. Those with GH[ge]2.5[mu]g/L and/or IGF-1[gt]ULN at 12m could switch treatment to PAS LAR 40mg/28d or OCT LAR 20mg/28d. Dose titration to PAS LAR 60mg/28d or OCT LAR 30mg/28d was permitted. All hyperglycemia-related adverse event (AE) terms were grouped. Efficacy data are from the first 6m after switching therapy; AEs are from any point in the extension.[br][bold]Results:[/bold] After completion of the core study, 81 and 38 patients switched to PAS LAR and OCT LAR, respectively. Response rates [95% CI] 6m after switching to PAS LAR and OCT LAR, respectively, were: GH[lt]2.5[mu]g/L [italic]and[/italic] normal IGF-1, 21.0% (17/81) [12.7, 31.5] and 2.6% (1/38) [0.1, 13.8]; normal IGF-1, 30.9% (25/81) [21.1, 42.1] and 7.9% (3/38) [1.7, 21.4]; GH[lt]2.5[micro]g/L, 43.2% (35/81) [32.2, 54.7] and 31.6% (12/38) [17.5, 48.7]. Tumor volume further decreased by a mean (SD) of 18.1% (17.7) and 12.3% (24.1) after patients switched to PAS LAR and OCT LAR, respectively. The safety profile of PAS LAR was similar to that of OCT LAR, with the exception of hyperglycemia-related events (60.5% vs 18.4%). Similar to the core study, the most common AEs (PAS LAR vs OCT LAR) were hyperglycemia (25.9% vs 7.9%), diarrhea (21.0% vs 15.8%), nasopharyngitis (16.0% vs 18.4%) and headache (18.5% vs 10.5%). Fasting plasma glucose and HbA[sub]1c[/sub] levels decreased to near normal levels within 3m in patients switched from PAS LAR to OCT LAR.[br][bold]Conclusions:[/bold] Switching treatment to PAS LAR allowed 21% of patients inadequately controlled with OCT LAR to achieve biochemical control. Only 2.6% of patients switched to OCT LAR achieved biochemical control. The safety profile of PAS LAR was similar to that of OCT LAR, with the exception of hyperglycemia-related events; hyperglycemia was reversible after PAS LAR discontinuation. PAS LAR may provide a treatment option for patients inadequately controlled with the standard medical therapy, OCT LAR.[br][br]Sources of Research Support: Novartis Pharmaceuticals.[br][br]Disclosures: MF: Researcher, Novartis Pharmaceuticals, Ipsen; Medical Advisory Board Member, Novartis Pharmaceuticals; Speaker, Ipsen, Chiasma. MS: Speaker, Novartis Pharmaceuticals. MDB: Consultant, Novartis Pharmaceuticals; Investigator, Novartis Pharmaceuticals. MG: Investigator, Novartis Pharmaceuticals; Speaker, Novartis Pharmaceuticals, Pfizer, Inc. KHR: Employee, Novartis Pharmaceuticals. MR: Employee, Novartis Pharmaceuticals. YC: Employee, Novartis Pharmaceuticals. AC: Investigator, Novartis Pharmaceuticals. Nothing to Disclose: PF, FG, C-CS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1733 203 1724 SUN-741 PO22-02 Sunday 1521 2012


1518 ENDO12L_SUN-742 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Pharmacokinetic and Pharmacodynamic Effects of Pasireotide (SOM230) Immediate Release and Long-Acting Release (LAR) in Rats and Mice Herbert A Schmid Novartis Pharma AG, Basel, Switzerland [bold]Introduction: [/bold]Pasireotide is a multireceptor-targeted somatostatin analogue with high affinity for somatostatin receptor subtypes sstr[sub]1,2,3[/sub] and sstr[sub]5[/sub] that has been developed for the treatment of neuroendocrine diseases. Two formulations of pasireotide have been developed: an immediate-release formulation for subcutaneous (sc) injection and a long-acting-release (LAR) formulation. The aim of this study was to establish the pharmacokinetic and pharmacodynamic properties of these formulations in rats and mice.[br][bold]Methods:[/bold] Male black 6 mice (n=4 each dose) were injected with pasireotide sc 5, 50 and 500 [micro]g/kg and pasireotide LAR 8, 80 and 320 mg/kg. Male Lewis rats (n=4 each dose) were injected with pasireotide sc 10, 15 and 30 [micro]g/kg and pasireotide LAR 8 and 80 mg/kg. Pasireotide plasma concentrations and glucose levels were measured for 24 h after pasireotide sc and for 28 and 49 days after pasireotide LAR administration in mice and rats, respectively.[br][bold]Results: [/bold]In mice, pasireotide sc 5, 50 and 500 [micro]g/kg resulted in peak plasma pasireotide levels of 2, 31 and 561 ng/mL 0.5 h after the injection; plasma levels rapidly fell below 1 ng/mL within 6 h. In rats, injection of much lower doses (10, 15, 30 [micro]g/kg) resulted in peak plasma pasireotide levels of 7, 9 and 22 ng/mL 0.5 h after the injection and remained [gt]1 ng/mL 6 h after injection. Administration of pasireotide LAR 8, 80 and 320 mg/kg in mice resulted in pasireotide levels of 14, 57 and 145 ng/mL, respectively, on day 1, reached peak values between days 7 and 14, and gradually reached 2, 17 and 172 ng/mL on day 28. Peak plasma levels were 15, 100 and 280 ng/mL, respectively. In rats, injection of 8 and 80 mg/kg pasireotide LAR resulted in peak plasma levels of 49 and 132 ng/mL on day 1, respectively. Peak values of 93 and 516 ng/mL were seen after 21[ndash]28 days, which gradually declined to 13 and 87 ng/mL after 49 days. Rapid elevations in plasma glucose levels were seen after injection of pasireotide sc in rats and mice. However, injection of pasireotide LAR resulted in only small and transient elevations in glucose levels in rats, but not in mice.[br][bold]Conclusions: [/bold]These results suggest that lower doses of pasireotide are required in rats than in mice to reach similar pasireotide plasma concentrations. In addition, pasireotide LAR appears to be associated with smaller and more transient elevations in glucose levels than pasireotide sc.[br][br]Sources of Research Support: Novartis Pharmaceuticals.[br][br]Disclosures: HAS: Employee, Novartis Pharmaceuticals. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1757 203 1725 SUN-742 PO22-02 Sunday 1522 2012


1519 ENDO12L_SUN-743 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Pegvisomant Alone or Combined with SSAs in Acromegaly: A Multicenter Retrospective Study Antonio Bianchi, Ferdinando Valentini, Patrizia Gargiulo, Maurizio Poggi, Roberto Baldelli, Marina Passeri, Laura Tilaro, Vincenzo Cimino, Marialuisa Appetecchia, Guido Tamburrano, Andrea Fabbri, Vincenzo Toscano, Alfredo Pontecorvi, Laura De Marinis Catholic University, Rome, Italy; S Camillo-Forlanini Hospital, Rome, Italy; University [quot]La Sapienza[quot], Rome, Italy; University [quot]La Sapienza[quot], 2nd Faculty, Rome, Italy; [quot]Regina Elena[quot] National Cancer Institute, Rome, Italy; University Tor Vergata, Rome, Italy Patients with acromegaly resistant to conventional drug treatment currently can advantage with GH-receptor antagonist pegvisomant (PEG) monotherapy or add pegvisomant to Somatostatin Analogs (SSA). No retrospective data are available about prognostic factors and efficacy for this different treatment regimens. This is a retrospective observational study including a total of 62 acromegalic patients (21 male and 41 female) treated for up to 7 years, 35 with PEG alone (Group A) and 27 with combination therapy (PEG+SSA) (Group B). No differences were found between the 2 groups in terms of age at diagnosis, micro- or macroadenomas, duration of disease, daily dose of PEG. In term of comorbidities, patients affected by OSAS were more frequent in Group B than A. No regrowth of adenomas or remnants were observed during the follow-up. Acromegalic patients with remnant of disease after unsuccesfully neurosurgery were more frequent in group B than group A (81.5 vs 48.5%, p[lt]0.05). Significant increase in GH levels and IGF-I levels at diagnosis and in the last control after PEG starting were observed in Group B. Group A showed a significant rate of controlled patient than group B (83.5 vs 55.5%, p[lt]0.05). In conclusion, our data showed that acromegalic patients treated with combination therapy were affected by a more aggressive disease and that combination treatment of these subjects does not seem more efficacy than PEG alone.[br][br]Nothing to Disclose: AB, FV, PG, MP, RB, MP, LT, VC, MA, GT, AF, VT, AP, LDM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1900 203 1726 SUN-743 PO22-02 Sunday 1523 2012


1520 ENDO12L_SUN-744 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Efficiency and Safety of Medical Therapy of Acromegaly Using High Doses of Sandostatin LAR Alexander V Dreval, Anna V Vinogradova, Irina V Trigolosova, Irena A Ilovayskaya Moscow Regional Research [amp] Clinical Institute Named by MF Vladimirsky, Moscow, Russian Federation Medical therapy by somatostatin analogues is one of the main options in acromegaly treatment. According to data from Moscow Region AcroRegistry, 79(61.7%) patients were treated by somatostatin analogues (Sandostatin LAR, S-LAR). Forty four of 79 (55.7%) patients got S-LAR as first-line therapy, and 35/79 (44.3%) patients received S-LAR as additional therapy (after neurosurgery n=20, radiation n=6, combined neurosurgery and radiation n=9). After 6-12 months of therapy by S-LAR 20 mg/4w 37 patients maintained the same dose, while 42 required dose 30 mg/4w, and after 6-12 months the dose was further increased to 40 mg/4w in 31 of these 42 patients. So, 37/79 (47%) patients received S-LAR 20 mg/4w (group20), 11/79 (14%) - 30 mg/4w (group30), and 31/79 (39%) - 40 mg/4w (group40). Micro-/macroadenomas (%) were 24/76, 18/82 and 16/84 in the groups respectively. Part of patients after previous treatment was 43,45,45% in groups 20,30,40 accordingly. Initial GH levels were 31[4.6;132] [amp] 28.6[19;176] [amp] 50[15;148] mMe/l and IGF-1(% from UNL) 151[102;188] [amp] 147[120;296] [amp]197[108;260] % in groups 20, 30, 40 respectively. After S-LAR therapy GH levels were 1.47[1;4] [amp] 3.6[1.7;6] [amp] 6.8[3.7;12] mMe/l and IGF-1 -25[-37;-12] [amp] -6[-23;87] [amp] 65[-6;135] % in corresponding groups. Both target GH and IGF-1 levels were achieved in 23/37 (62.6%), 6/11 (54.5%) and 10/31 (32.2%) patients from groups 20, 30 and 40, respectively. In addition, at least one target value GH or IGF-1 was reached in another 10/33 (30.3%), 4/11 (36.4%) and 9/31 (29%) patients in these treatment groups. Increase of the S-LAR dose to 30-40 mg allowed to achieve full control of GH and IGF-1 levels in other 16 patients. The frequency and/or intensity of adverse events were not increased at dose rising.[br]Thus, among our group of not selected acromegalic patients receiving S-LAR achievement of one target level was observed at 62 of 79 (78.5%) patients, and full biochemical control of acromegaly - at 39 of 79 (49.4%) patients. For effective acromegaly control more than half of patients needed doses of 30 mg and above. Initial GH/IGF-1 levels as well as previous treatment were poor predictors of efficiency of S-LAR therapy. The use of high doses of S-LAR (to 40 mg) was safe, well tolerated, and increased amount of patients with acromegaly who have reached full biochemical control. Probably, it is reasonable to conder possibility of the further augmentation of S-LAR doses (to 60 mg) for better acromegaly control.[br][br]Disclosures: AVD: Speaker, Lilly USA, LLC, Novo Nordisk, Novartis Pharmaceuticals, Ipsen. IAI: Speaker, Abbott Laboratories, Ipsen, Novartis Pharmaceuticals. Nothing to Disclose: AVV, IVT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 668 203 1727 SUN-744 PO22-02 Sunday 1524 2012


1521 ENDO12L_SUN-745 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Acute Octreotide Suppression Test for Evaluation of Subsequent Therapy with Somatostatin Analogues in Newly Diagnosed Acromegaly Inga Balcere, Ieva Strele, Janis Klovins, Valdis Pirags University of Latvia, Riga, Latvia; Riga Stradin[scaron] University, Riga, Latvia; Latvian Biomedical Research and Study Center, Riga, Latvia [bold]Background[/bold]. It has been reported that primary treatment with (SSA) is effective in up to 60% of acromegaly patients, but predictive value of the acute octreotide suppression test (OST) for selection of patients with good response to depot SSA in long term treatment remains controversial.[br][bold]Patients and Methods[/bold]. 12 medical therapy naive patients (mean age 44 y, female 75%) with active acromegaly were included in prospective study. After administration of 100mcg octreotide the baseline and hourly GH measurements were taken for following 6 hours. GH response to the OST was defined as the per cent reduction of GH from the baseline. Subsequently 6 patients received the long-acting SSA lanreotide ([italic]Somatuline Autogel[/italic]), 120mg every 28 d and 6 patients received long-acting SSA octreotide ([italic]Sandostatin LAR)[/italic] 20-40mg every 28 d for 3-24 month treatment period. GH and IGF1 were measured at baseline and during the treatment period.[br][bold]Results[/bold]. 58% of patients (7 out of 12) showed good response ([gt]50% GH decrease) during the OST. 42% of them (3/7) reached target GH[lt] 2ng/ml during the subsequent SSA therapy and only one patient (14%) achieved normalization of IGF1. 25% of patients (3 out of 12) showed moderate 30-50% GH decrease during the OST and only one reached normal IGF1 after administration of long-acting SSA. Despite that 17% of patients (2 out of 12) had only small [lt]30% GH decrease during the OST, they reached the treatment target (GH[lt] 2ng/ml) during the subsequent depot SSA therapy.[br][bold]Conclusion[/bold]. Acute OST has poor predictive value for effectiveness of subsequent long-acting SSA therapy and should not replace at least 3 month trial with depot SSA.[br][br]Sources of Research Support: University of Latvia PhD fellowship, European Sopcial Foundation.[br][br]Nothing to Disclose: IB, IS, JK, VP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1608 203 1728 SUN-745 PO22-02 Sunday 1525 2012


1522 ENDO12L_SUN-746 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) Efficacy and Safety in the Somatuline[reg] Depot (lanreotide) Injection for Acromegaly (SODA) Study Conducted in the U.S.: Overall Year 1 Data and Comparison of Home Injection Versus Office Injection Roberto Salvatori, Whitney Woodmansee, Cathryn Clary, Stephen Chang, Bert Bakker Johns Hopkins University School of Medicine, Baltimore, MD; Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA; Ipsen Inc, Brisbane, CA [bold]Introduction:[/bold] The Somatuline[reg] Depot (lanreotide) Injection for Acromegaly (SODA) study is an ongoing, multi-center, observational, post-marketing study of Somatuline Depot (SD)-treated patients (pts) with acromegaly, reflecting [ldquo]real world[rdquo] SD experience. This is the first efficacy/safety analysis comparing patients who were administered SD by self (Group 1), partner (Group 2), healthcare provider (HCP) (Group 3), or mixed methods (Group 4), conducted in the United States.[bold] Methods:[/bold] Pts enrolled were clinically diagnosed with acromegaly and administered SD (new pts or pts switched from prior treatments). Pts were enrolled at any time from starting SD. All demographics, efficacy data, adverse events (AE), serious adverse events (SAE) were analyzed. [bold]Results: [/bold]By Jan 5, 2012, of 166 pts enrolled in the SODA database, 49% were female, average age was 50.1 years, 28 (17 %) self-injected, 43 (26 %) were administered by partner, 60 (36 %) were administered by HCP and 35 (21%) received mixed injection methods. Most pts[apos] initial SD dose was 90 mg (71%); 80% had undergone pituitary surgery, 26% were previously na[sub]i[/sub]ve and 49% were previously treated with octreotide LAR. Of 67 pts with IGF-1 levels measured at 1 year after enrollment, mean IGF-I (Std Dev) of all and 4 groups were, respectively: 271(197), 229(79), 244(160), 282(211), 306(258) ng/mL. The percentage of pts with serum IGF-1 normal or low for age were 73%, 85%, 86%, 65%, 65%, respectively. 35 pts had GH measured at 1 year after enrollment. The percentage of pts with fasting GH level [lt]= 2.5 ng/mL were 77%, 83%, 83%, 75%, 73%, respectively. 152 pts[apos] follow-up data were included in the safety analysis. 96 (63%) reported at least one AE. The 3 most commonly reported targeted Aes (TAEs) were headache (22%), diarrhea (20%) and abdominal pain (19%). Injection site reactions were reported more frequently in self-injectors. 15 (10%) pts reported 40 SAEs deemed unrelated to SD; 21 pts discontinued the study early for the following reasons: death unrelated to SD (2), AE (2), and other (17).[bold] Conclusions:[/bold] At 1 year, a majority of pts had normal or low IGF-1 levels reflecting biochemical control of acromegaly, regardless of whether pts self-injected or received injection by partner or HCP. All injection methods for SD have acceptable TAE profiles. In this open-label observational study 43% of pts utilized self-injection or partner injection, which appeared similarly efficacious, and as safe as injection by HCP.[br][br]Sources of Research Support: Ipsen.[br][br]Disclosures: RS: Advisory Group Member, Ipsen, Novartis Pharmaceuticals; Research Funding, Ipsen, Novartis Pharmaceuticals, Pfizer, Inc. WW: Consultant, Eli Lilly & Company; Advisory Group Member, Eli Lilly & Company; Researcher, Ipsen; Advisory Group Member, Ipsen; Researcher, Pfizer, Inc. CC: Stockholder, Pfizer, Inc., Ipsen; Owner, Ipsen; Employee, Ipsen. SC: Employee, Ipsen; Stockholder, Ipsen. BB: Stockholder, Ipsen; Stockholder, Ipsen; Employee, Ipsen. 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 777 203 1729 SUN-746 PO22-02 Sunday 1526 2012


1523 ENDO12L_SUN-747 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) The Effect of Somatostatin Analogs on Serum Calcium and 25 OH Vitamin D Concentrations in Patients with Acromegaly: A Retrospective Study of 119 Patients Lisa B Nachtigall, Shirin Attarian, Maya Barake, Nicholas A Tritos, Anne Klibanski, Karen K Miller Massachusetts General Hospital/Harvard Medical School, Boston, MA [bold]Background[/bold]: Little is known about the effect of growth hormone excess or the impact of somatostatin analog (SSA) therapy on calcium and mineral metabolism in acromegaly. The goals of this study were to determine: 1) 25 OH vitamin D (25OHD) and calcium (Ca) levels in patients with acromegaly and their association with IGF-1 levels and 2) whether SSA therapy affects calcium and 25 OH D levels. [bold]Design[/bold]: 119 patients with acromegaly in whom 25OHD levels were available were studied (mean age 54[plusmn]1 (SEM) yr; M/F: 39/80). The mean IGF-I index calculated as IGF-I value/maximal normal value for assay was 0.75[plusmn]0.05 SEM. Ca and 25OHD levels were compared between patients receiving (N=31) vs. not receiving (N=73) SSA. Medical therapy in the latter group included pegvisomant and/or cabergoline; IGF-I levels did not differ compared to the SSA group. Calcium and 25OHD levels were also evaluated longitudinally in a subgroup with serial levels prior to and sequentially during short-term (mean 3 months, range 1-5) and long-term (mean 49 months, range 7-180) SSA administration. [bold]Results[/bold]: Calcium and albumin levels were normal pretreatment and throughout longitudinal follow up. Calcium correlated with IGF-I (N=119, R=0.24, p=0.017). In patients with comparable IGF-I levels and season of testing, there was no difference in serum 25OHD levels in patients who were (N= 31) or were not receiving SSA (N= 73)(35[plusmn]1 ng/ml vs. 35[plusmn]2 (SEM), p=0.96). In the longitudinal subset (N=16), serum Ca levels decreased significantly, but transiently, in patients receiving short-term SSA (pretreatment 9.9[plusmn]0.1 mg/dl vs. short-term SSA 9.5[plusmn] 0.1, p=0.004). After long-term administration of SSA, Ca increased compared to levels on short-term therapy (9.8[plusmn]0.1 mg/dl vs. 9.5[plusmn]0.1, p=0.017) and were unchanged compared to baseline. The change in IGF-I levels was not a determinant of change in calcium levels at any time point. Neither PTH nor 25OHD changed with short-term SSA administration. [bold]Conclusions[/bold]: Calcium is positively associated with IGF-I levels in patients with acromegaly. There is a transient decrease in Ca with short-term SSA use. The SSA acute effect on Ca does not appear to be mediated by albumin, 250HD or PTH and resolves with long-term SSA treatment. In patients receiving long-term SSA therapy, vitamin D levels are normal and similar to those of patients receiving other therapies, suggesting that long-term SSA therapy does not affect serum vitamin D concentrations.[br][br]Sources of Research Support: Investigator Initiated Grant from Ipsen.[br][br]Disclosures: LBN: Principal Investigator, Ipsen. NAT: Consultant, Pfizer, Inc. Nothing to Disclose: SA, MB, AK, KKM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 534 203 1730 SUN-747 PO22-02 Sunday 1527 2012


1524 ENDO12L_SUN-748 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) A Prospective Study of Two-Year High-Dose Cabergoline Therapy for Acromegalic Patients with Post-Operative Residual Tumor Msami Ono, Nobuhiro Miki, Atsuhiro Itihara Endocrinology and Hypertension, Shinjuku-ku, Tokyo, Japan [bold]Context[/bold]: Unlike remarkable efficacy in prolactinoma, cabergoline is much less effective in acromegaly. However, this evidence was obtained mostly from clinical studies that employed a relatively low dosage of cabergoline for a short period of [lt] 6 months.[br][bold]Objective[/bold]: This prospective study was conducted to treat acromegalic patients with individualized high-dose cabergoline therapy for a longer duration of 2 years.[br][bold]Methods[/bold]: Patients were 50 acromegalics (38 women and 12 men) with post-operative residual tumor. Immunohistochemically, 33 cases were GH adenomas and the remaining 17 cases were GH-PRL cell adenomas. No patients had received prior radiotherapy. Cabergoline was started at a standard dosage and schedule used for prolactinoma treatment. Its dose was incrementally adjusted at 3-month intervals based upon the degree of decline in serum IGF-I, which was measured 2-3 months after the closest dose-escalation day. The highest daily and weekly doses were set at 3 mg/day and 9 mg/week, respectively. Eight resistant patients agreed to receive doses greater than the 9 mg/wk for a limited duration of a half or 1 yr after.[br][bold]Results[/bold]: Cabergoline suppressed both elevated IGF-I and GH serum levels in all patients. Decline in IGF-I into the age/gender-adjusted normal range was achieved in 37 (74%) patients and decrease in serum GH below 2.5 mg/L occurred in 36 (72%) patients. Serum IGF-I began to be normalized 4 months after cabergoline and thereafter, its normalization rate increased linearly, reaching 38 % at 1 year and 74% at 2 years. Tumor type-specific normalization of IGF-I was 70% for GH cell adenomas and 82% for GH-PRL cell adenomas. The normalization rate of IGF-I was related to pretreatment levels; 100 % in 20 patients with 293-488 mg/L, 85% in 20 subjects with 489-732 mg/L, and zero% in 10 cases with 733-1700 mg/L. Thirty-seven (88%) of the 42 patients treated with 1-9 mg/wk doses could normalize their IGF-I levels, while any of the remaining 8 patients treated with the highest 12-21 mg/wk doses could not normalize serum IGF-I levels and cabergoline was diminished to a minimum effective dose that produced a maximum effect on IGF-I. There were no adverse effects of cabergoline including cardiac valvulopathy.[br][bold]Conclusion[/bold]: By introducing a long-term suppressive therapy with the use of a sufficient dosage, cabergoline likely becomes able to control IGF-I production in a majority of acromegalic patients with a mild to moderate degree of residual tumor activity.[br][br]Nothing to Disclose: MO, NM, AI 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 1527 203 1731 SUN-748 PO22-02 Sunday 1528 2012


1525 ENDO12L_SUN-749 POSTER SESSION: Novel Medical Treatments for Pituitary Tumors (1:30 PM-3:30 PM) The Glucagon Stimulation Test (GST) in Assessing GH Reserve and Adrenal Function in Adults with Non-Pituitary Hypothalamic Disease and Pituitary Non-Functioning Adenomas with Low Serum IGF-I Levels KCJ Yuen, L Katznelson, SA Rhoads, MH Gurel, O Chu, V Corazzini, R Salvatori, DM Cook, BMK Biller Oregon Health [amp] Science University, Portland, OR; Stanford University School of Medicine, Stanford, CA; Massachusetts General Hospital, Boston, MA; The Johns Hopkins School of Medicine, Baltimore, MD [italic]Context:[/italic] The GHRH-ARG test is a well-accepted test for assessing adult GH deficiency (GHD). Previous studies using the GHRH-ARG test have reported preservation of somatotroph responsiveness in patients with recent hypothalamic causes of GHD (1) making this test unreliable in these patients. However, the utility of the GST in such patients is unclear. Thus, we assessed the GST in inducing GH [amp] cortisol secretion in patients with non-pituitary hypothalamic disease (HTD) [amp] pituitary non-functioning adenomas (NFA) with low serum IGF-I levels.[br][italic]Methods:[/italic] Patients referred to 4 Endocrine Testing Units with HTD [[italic]n[/italic]=19, 5M, prior irradiation for non-pituitary brain tumors ([italic]n[/italic]=8), hypothalamic tumors ([italic]n[/italic]=7), childhood-onset GHD without pituitary tumors ([italic]n[/italic]=4), age 34.4 [plusmn] 2.8 yr, BMI 28.5 [plusmn] 1.7 kg/m[sup]2[/sup]] [amp] NFA [[italic]n[/italic]=78, 28M, 40 macro- [amp] 38 microadenomas, no history of cranial irradiation, age 50.6 [plusmn] 1.5 yr, BMI 30.7 [plusmn] 0.9 kg/m[sup]2[/sup]] suspected of GHD [amp] adrenal insufficiency (AI) were studied. Surgery was performed in 2 [amp] 42 patients with HTD [amp] NFA, respectively. All subjects underwent fixed-dose GSTs (1 mg [amp] 1.5 mg IM glucagon for 90 kg [amp] [gt] 90 kg body weight, respectively). Cutoff GH [amp] cortisol levels were set at 3 [micro]g/L [amp] 18 [micro]g/dL, respectively.[br][italic]Results:[/italic] There were no differences in BMI and gender distribution between groups, but HTD patients were younger ([italic]P[/italic][lt]0.001). In the HTD group, fasting ([italic]P[/italic][lt]0.01) [amp] nadir ([italic]P[/italic][lt]0.05) glucose levels were lower than the NFA group, but not peak [amp] [Delta]glucose levels. IGF-I SDS (-1.9 [plusmn] 0.2 vs -2.0 [plusmn] 0.1; [italic]P[/italic]=NS), basal, peak [amp] [Delta]GH, basal ACTH, basal [amp] [Delta]cortisol were comparable in the 2 groups, but peak cortisol levels were higher in the HTD group ([italic]P[/italic][lt]0.05). Fewer HTD patients reported nausea (16 % vs 33%; [italic]P[/italic][lt]0.01), [amp] failed the GST for GH (47% vs 65%; [italic]P[/italic][lt]0.05) [amp] cortisol (11% vs 62%; [italic]P[/italic][lt]0.0001) than NFA patients. No patient with HTD vomited, whereas 2 patients with NFA vomited at 90 mins during the GST.[br][italic]Conclusion:[/italic] Unlike the GHRH-ARG test, the GST induces comparable GH responses in patients with hypothalamic [amp] pituitary diseases with low serum IGF-I levels, [amp] may also induce cortisol release in patients with HTD. Thus, the ability of glucagon to induce GH [amp] cortisol secretion is comparable regardless of whether the injury is at the hypothalamus or pituitary level. Further studies are needed to establish the GH [amp] cortisol GST cutoff levels in comparison to the gold standard stimulation test (i.e. insulin tolerance test) in assessing for GHD in patients with HTD.[br][br]1. Darzy KH, et al. The usefulness of the combined growth hormone (GH)-releasing hormone [amp] arginine stimulation test in the diagnosis of radiation-induced GH deficiency is dependent on the post-radiation time interval. J Clin Endocrinol Metab 2003; 88:95-102.[br][br]Sources of Research Support: KCJY has received research grants from Novo Nordisk. BMKB has received research grants and consulting honoraria from Novo Nordisk. LK, RS and DMC have received consulting honoraria from Novo Nordisk. SAR, MHG, OC and VC all have nothing to declare.[br][br]Disclosures: LK: Investigator, Novartis Pharmaceuticals, Corcept Therapeutics; Research Funding, Pfizer, Inc., Ipsen. Nothing to Disclose: KCJY, SAR, MHG, OC, VC, RS, DMC, BMKB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 440 203 1732 SUN-749 PO22-02 Sunday 1529 2012


1526 ENDO12L_SUN-LB1 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Beta-Cell Function in Latent Autoimmune Diabetes in Adults Treated with Linagliptin vs Glimepiride: Exploratory Results from a 2-Year Double-Blind Randomized Controlled Study Odd Erik Johansen, Bernhard Boehm, Valdemar Grill, Peter A Torjesen, Sudipta Bhattacharya, Sanjay Patel, Kristiane Wetzel, Hans-Juergen Woerle Boehringer Ingelheim, Ingelheim, Germany; Ulm University, Ulm, Germany; Norwegian University of Science and Technology, Trondheim, Norway; Oslo University Hospital - Aker, Oslo, Norway; Boehringer Ingelheim, Biberach, Germany; Boehringer Ingelheim, Bracknell, UK [bold]Background[/bold]: Latent autoimmune diabetes in adults (LADA) is associated with a more rapid decline in beta-cell function compared to common type 2 diabetes (T2D). Presently, no treatment modality is drug of choice in LADA. We compared the impact of treatment with the DPP-4 inhibitor linagliptin (5 mg/day) and the sulphonylurea glimepiride (1-4 mg/day) on beta-cell function in patients retrospectively identified with LADA who had insufficient glycemic control despite metformin therapy in a 2-year study.[br][bold]Methods[/bold]: Patients were classified as LADA if one or more of the autoantibodies assessed (GAD65, ICA, IA-2A, IAA) were present at baseline or any on-treatment visit. GAD was assessed using RIA methodology (cut-off 0.05 [sensitivity 82%/specificity 98.89% in the Diabetes Autoantibody Standardization Program 2010]).[br][bold]Results[/bold]: The study cohort comprised 1519 patients (16 countries) with assumed common T2D. The prevalence of LADA was 7.8% (n=118). GAD65 was the most prevalent autoantibody (6.5%) whereas ICA (0.3%), IA-2A (1.2%), and IAA (0.2%) were rare. The proportion of patients with at least 2 positive antibodies was 0.4%. Baseline characteristics in GAD65+ LADA patients treated with linagliptin (n=65) or glimepiride (n=53) were fairly well balanced (respective age 59/63 years, BMI 30.3/31.7 kg/m[sup]2[/sup], and diabetes duration [gt]5 years 62%/59%). Among GAD65+ patients with C-peptide measurements available, those treated with linagliptin preserved C-peptide significantly better than those treated with glimepiride over a 2-year trajectory. At 28, 52, and 104 weeks, changes from baseline in the linagliptin group were +96 (baseline C-peptide 821, n=21), +143 (baseline C-peptide 835, n=14), and +202 (baseline C-peptide 944, n=9), all [italic]P[/italic][lt]0.001 versus baseline; corresponding changes in the glimepiride group were -105 (baseline C-peptide 1326, n=17), -179 (baseline C-peptide 1425, n=14), and -29 (baseline C-peptide 1374, n=9), [italic]P[/italic][lt]0.01 vs linagliptin at 28 and 52 weeks. HbA1c reductions were of similar magnitude in the linagliptin and glimepiride groups (changes from baseline at 28, 52, and 104 weeks were -0.25, -0.49, and -0.41 in the linagliptin group and -0.75, -0.52, and -0.49 in the glimepiride group).[br][bold]Conclusion:[/bold] Treatment with linagliptin or glimepiride in LADA could have differing impacts on long-term beta-cell function. Further research exploring the observed potential modulating impact of linagliptin on LADA is ongoing.[br][br]Sources of Research Support: This study was supported by Boehringer Ingelheim.[br][br] SB: Employee, Boehringer Ingelheim. SP: Employee, Boehringer Ingelheim. KW: Employee, Boehringer Ingelheim. H-JW: Employee, Boehringer Ingelheim. Nothing to Disclose: OEJ, BB, VG, PAT 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3093 204 1733 SUN-LB1 PO02_LB Sunday 1530 2012


1527 ENDO12L_SUN-LB2 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Abiraterone Acetate Added to Physiologic Hydrocortisone Dosing Safely Controls Androgen Excess in Adult Women with Classical 21-Hydroxylase Deficiency Richard J Auchus, Elizabeth Osborne, Gary D Hammer, Alice Y Chang, Carole Ramm, David Madrigal, Margaret Yu University of Michigan, Ann Arbor, MI; UT Southwestern Medical Center, Dallas, TX; Janssen Research and Development, Los Angeles, CA The most common form of congenital adrenal hyperplasia is 21-hydroxylase deficiency (21OHD). Treatment for 21OHD consists of glucocorticoid and mineralocorticoid replacement. In women, overtreatment with glucocorticoids to control androgen excess is common, leading to cushingoid side effects, including weight gain, depression, easy bruising, skin thinning, overuse injuries, bone disease and poor quality of life. Better treatments to control androgen excess in 21OHD are needed. Abiraterone acetate (AA) is a potent, orally-active inhibitor of cytochrome P450c17 (CYP17A1), which impairs androstenedione (AD) and testosterone (T) synthesis. AA is approved for treatment of prostate cancer in combination with prednisone, to prevent accumulation of 21-hydroxysteroids, which cause hypertension and hypokalemia. We hypothesized that AA added to physiologic hydrocortisone would improve androgen excess in women with 21OHD without causing hypertension and hypokalemia.[br]Key inclusion criteria for this Phase I open-label, multiple-dose, intra-subject sequential dose-escalation study are classical 21OHD genotype and serum AD [gt]1.5x normal ([gt]345 ng/dL) while taking physiologic hydrocortisone replacement and fludrocortisone. AA is taken as a single morning dose 1 h prior to food and hydrocortisone on days 1-6. Blood and urine samples before any medications are obtained on study days 1, 6, 7, and 8, with a time course after the final dose on day 6. The primary endpoint is AD, mean of days 6 [amp] 7. Secondary endpoints include 17-hydroxyprogesterone, total T, urine androsterone and etiocholanolone (T metabolites), plus electrolytes, blood pressure, and safety labs.[br]For the first 3 adult women with 21OHD treated with AA 100 mg/d, serum AD decreased from a mean baseline of 610 ng/dL to mean day 6 [amp] 7 of 150 ng/dL (75% from baseline) and to normal ([lt]230 ng/dL) in 2 of 3. After the final dose, AD fell to a mean nadir of 47 ng/dL by 8 h (92% from baseline). T also decreased from 108 to 27 ng/dL (75% from baseline) on day 6 and 85% from baseline after the final dose. Urine T metabolites fell in parallel to T. AA was safe and well tolerated. No adverse events of hypertension or hypokalemia were observed.[br]We conclude that AA added to replacement hydrocortisone and fludrocortisone markedly lowers serum AD, testosterone, and urine T metabolites without hypertension or hypokalemia after 6 daily doses in adult women with 21OHD. Updated results for all participants will be presented.[br][br]Sources of Research Support: Janssen Pharmaceuticals.[br][br]Nothing to Disclose: RJA, EO, GDH, AYC, CR, DM, MY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3060 204 1734 SUN-LB2 PO02_LB Sunday 1531 2012


1528 ENDO12L_SUN-LB3 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Comparison of the Effect of High Calorie Breakfast Diet vs High Calorie Dinner Diet on Weight Loss, Ghrelin, Lipids and Appetite Scores in Obese Non Diabetic Women Daniela Jakubowicz, Mona Boaz, Yosepha Bar-Dayan, Julio Wainstein Tel Aviv University, Holon, Israel; Tel Aviv University, Holon, Israel [bold]Background[/bold]: Recently we have shown that compared to low carbohydrate (CH) diet, an isocaloric diet with addition of high calorie breakfast (B) that also included dessert, promoted sustained weight(W) loss(WL) and prevented W regain by reducing diet-induced compensatory changes in hunger, cravings and ghrelin suppression. Direct effects of meal timing (morning vs.evening increased calories) were not tested.[br][bold]Objective: [/bold]To search whether a change in meal timing by increasing calories in the morning vs in the evening has a differential impact on WL, ghrelin suppression, lipids and appetite scores. Our hypothesis is that comparing with high calorie dinner (D) + reduced B (HCDd), the high calorie B + reduced D (HCBd), would enhance WL, ghrelin suppression and appetite scores.[br][bold]Methods[/bold]: 73 obese women (BMI 32.3[plusmn]2.0kg/m2), aged 46[plusmn]6 years, were randomized to two isocaloric (1400 kcal) weight loss diets during 12 weeks(Wk). Both diets had the same composition but differ in the meal timing distribution[br]1)HCBd consisted in large B with 700Kcal(Kc)with % of CH;protein;fat: 50:30:20%; lunch (500Kc,20:45:25%), and small D (200 Kc,13:40:47%).[br]2)HCDd consisted in low calorie B (200Kc,13:40:47%),lunch(500Kc,20:45:25%), and large D (700Kc,50:30:20%).[br]Anthropometric measures were assessed every 4Wk. Fasting glucose, insulin, ghrelin, lipids, OGTT, craving scores as well as B and D challenge were performed at baseline and at Wk12[br][bold]Results[/bold]: After Wk12 intervention, the HCBd group (g) lost 8.7[plusmn]1.4 kg vs 3.48[plusmn]2.3 kg the HCDd g. In comparison to the HCDd g, the HCBd g showed 61% greater WL, P[lt]0.05,a 35% greater reduction in waist circumference (P[lt]0.06) and a 17% greater reduction in percent body fat (P=NS). Ghrelin levels were reduced after B by 46.2% in the HCBd group and 18.5% after D in the HCDd g(P[gt]0.005). Satiety was significantly improved and hunger and craving scores significantly reduced in the HCBd vs HCDd g (P[gt]0.005). Mean serum triglyceride levels decrease by 44% (from184.2[plusmn]16.6 to102.2[plusmn]7.7mg/dl) in the HCBd g while increase 6.3% (from180.5[plusmn]20.6 to192.4[plusmn]17.5mg/dl) in the HCDd g(P[gt]0.005). Total cholesterol (C), HDL-C and LDL-C, did not differ between the groups.[br][bold]Conclusion[/bold]: Isocaloric WLdiets with different meal timing, differently influence WLrate, ghrelin, appetite and lipid levels. HCBd with reduced intake at D result in enhanced WL, increased ghrelin suppression and might be useful alternative for management of obesity.[br][br]Nothing to Disclose: DJ, MB, YB-D, JW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3008 204 1735 SUN-LB3 PO02_LB Sunday 1532 2012


1529 ENDO12L_SUN-LB4 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) 1,5-Anhydroglucitol (1,5-AG) Is Strongly Associated with Neonatal Birth Weight across All Trimesters in Pregnancy Complicated by Diabetes Lorena Alarcon-Casas Wright, Irl B Hirsch, Ted A Gooley, Shani S Delaney, Zane Brown University of Washington, Seattle, WA; University of Washington and Fred Hutchinson Cancer Research Center, Seattle, WA; University of Washington, Seattle, WA Low levels of 1,5-AG, a biomarker of short-term glucose control, are associated with postprandial hyperglycemia in the non-pregnant diabetic population. Postprandial hyperglycemia and hemoglobin A1C (A1C) during pregnancy are correlated with neonatal birth weight (NBW). To determine whether 1,5-AG is associated with NBW during pregnancy complicated by diabetes, we obtained simultaneous measurements of 1,5-AG and A1C, every 4-8 weeks throughout pregnancy until delivery in 101 women with diabetes: Type 1 DM (T1)= 42, Type 2 DM (T2)= 31, Gestational Diabetes (GD)= 28. NBWs were obtained from the infants[apos] medical records and evaluated by standardized z-scores, controlling for gestational age at delivery and gender. Generalized estimating equations were used to evaluate the association of 1,5-AG, A1C, and their ratio (1,5-AG/A1C) with NBW z-scores, accounting for within-subject correlation due to multiple NBWs within individual women. Means for A1C (%), 1,5-AG ([micro]g/ml), and NBW z-scores were 6.6, 3.9 and 0.56; 6.5, 6.0 and 0.32; and 5.9, 7.1 and 0.48, in T1, T2 and GD, respectively. A negative association was found between NBW z-scores and 1,5-AG and 1,5-AG/A1C among all women; p[lt]0.0001, regardless of type of diabetes and trimester. The association was non-linear, and as 1,5-AG increased, the decrease in NBW z-scores leveled off. A non-linear association was also observed between A1C and NBW z-scores; p=0.008. However, with the non-linear function of 1,5-AG in the model for NBW, adding the linear component of A1C did not improve the model; p=0.77. Oppositely, adding the linear component of 1,5-AG to the model containing the non-linear function of A1C led to a statistically significant change; p=0.0001. There was no evidence to support the hypothesis that the associations were different by type of diabetes or by trimesters: statistical tests of interaction yield p=0.75, 0.80 and 0.45 when comparing the associations between groups. Similarly, the associations by trimesters were qualitatively non-significant: p=0.85 for both 1[sup]st[/sup] and 2[sup]nd [/sup]and 1st and 3[sup]rd [/sup]trimesters; and 0.48 for 2[sup]nd[/sup] and 3[sup]rd[/sup] trimesters.[br]Conclusion: 1,5-AG and 1,5-AG/A1C offer additional and superior information on NBW in all types of diabetes and across all trimesters, beyond A1C alone. By providing more information on glycemic exposure, 1,5-AG is a promising adjunct in the management of pregnancy complicated by diabetes.[br][br]Sources of Research Support: The Department of Obstetrics and Gynecology/Maternal-Fetal Medicine, University of Washington.[br]NIH grant T32DK0007247.[br][br]Nothing to Disclose: LA-CW, IBH, TAG, SSD, ZB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3035 204 1736 SUN-LB4 PO02_LB Sunday 1533 2012


1530 ENDO12L_SUN-LB5 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Impact of Obstructive Sleep Apnea on Glucose Tolerance and Appetite Regulation: Sex Differences Karla A Temple, Fanny Delebecque, Rachel Leproult, Lisa L Morselli, Harry Whitmore, Babak Mokhlesi, David A Ehrmann, Eve Van Cauter University of Chicago, Chicago, IL; University of Chicago, Chicago, IL Obstructive sleep apnea (OSA) is an independent risk factor for metabolic dysfunction, and is more prevalent in men than women. We sought to determine if there are sex differences in the impact of OSA on insulin, glucose, pancreatic polypeptide (PP), leptin, glucose tolerance, insulin sensitivity, beta-cell responsiveness and diabetes risk.[br]143 overweight/obese men and women had a polysomnogram and a fasting 2h 75g OGTT during which glucose, insulin, C-peptide, leptin, and PP were assayed. OSA severity was assessed by the apnea-hypopnea index (AHI). Fasting insulin resistance (IR) was estimated by HOMA-IR. IR during the OGTT was assessed by the area under the curve of HOMA-IR, and insulin sensitivity was assessed by the Matsuda index. Beta-cell response was assessed by the insulinogenic index and by C-peptide levels; diabetes risk by the oral disposition index (DI; Matsuda index x insulinogenic index). Comparisons were adjusted for age, BMI, and race.[br]40 men and 37 women had OSA (73% vs 42%; p[lt]0.001). Prediabetes was present in 27% of women but 45% of men (p[lt]0.05) with OSA. OSA was associated with higher fasting glucose (p[lt]0.001) and higher PP levels (p[lt]0.0001) in women, but not men. During the OGTT, glucose tolerance was more impaired in the presence of OSA, without significant sex differences. The adverse impact of OSA on fasting IR was similar in men and women. In contrast, during the OGTT, increased severity of OSA tended to be associated with a larger decrease in insulin sensitivity (p[lt]0.09) and a greater increase in IR (p[lt]0.05) in men than in women. With increasing AHI, men had an increase in insulinogenic index (r[sup]2[/sup]=0.21;p=0.003), while women had a decrease in insulinogenic index (r[sup]2[/sup]=0.20; p=0.006). Increasing AHI was associated with a greater decrease in oral DI in women than in men (women: r[sup]2[/sup]=0.43; p[lt]0.0001, men: r[sup]2[/sup]=0.17;p[lt]0.002). Increasing AHI was also associated with an increase in PP levels in women, but a decrease in PP levels in men. Higher leptin levels were associated with increased AHI in women but not in men (p[lt]0.03).[br][bold]Conclusions[/bold]: OSA leads to an increase in appetite-suppressing hormones in women. OSA is associated with decreased glucose tolerance in both men and women, but the cause differs by sex. OSA is associated with greater reductions in insulin sensitivity in men than in women. However, the adverse impact of OSA on beta-cell responsiveness is larger in women than in men, resulting in an overall greater risk of diabetes in women.[br][br]Sources of Research Support: This work was supported by NIH grants MO1-RR-00055, P50-HD057796, R01-HL75025, PO1 AG-11412, P60-DK020595.[br][br]Nothing to Disclose: KAT, FD, RL, LLM, HW, BM, DAE, EV-C 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3077 204 1737 SUN-LB5 PO02_LB Sunday 1534 2012


1531 ENDO12L_SUN-LB6 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Optimising Healthy Gestational Weight Gain in Women at High Risk of Gestational Diabetes: A Randomised Controlled Trial Helena Jane Teede, Catherine Lombard, Cheryce Harrison Monash University, Melbourne, VIC, Australia; Southern Health, Melbourne, VIC, Australia Excessive gestational weight gain is common with significant health implications including increased risk of gestational diabetes (GDM). In the longer term, postpartum weight retention increases health risks including diabetes. We aimed to optimise healthy gestational weight gain in women at increased risk of gestational diabetes mellitus (GDM)using a low intensity, low cost lifestyle intervention, early in pregnancy. In a randomised controlled trial, 228 pregnant women at risk of developing GDM were recruited from a hospital setting. Women allocated to the intervention received a four session behavioural change lifestyle program, whilst the control group received a pamphlet with pregnancy and lifestyle information only. End point data was collected at 12-15 weeks and 26-28 weeks gestation and 6 weeks postpartum. Endpoints included anthropometrics (weight and height), physical activity assessment(Yamax pedometer), questionnaires and GDM screening (28 weeks). The study was powered for change in weight, not for difference in GDM incidence. At baseline the mean age [31.7(4.5) and 32.4(4.7) years] and BMI [30.3(5.9) and 30.4(5.6)kg/m2] were similar between control and intervention groups, respectively. By 28 weeks gestation there was a significant difference in weight gain between control and intervention groups [6.9(3.3) versus 6.0(2.8)kg, p[lt]0.05]. The proportion of women exceeding Institute of Medicine recommendations for gestational weight gain was significantly reduced in the intervention group compared to controls, with results most marked in overweight women (17% vs 55%). Physical activity levels declined overall by 28 weeks (p[lt]0.01), however the intervention group had a 20% higher step count compared to controls [5203(3368) vs 4140(2420) steps/day, p[lt]0.05]. GDM prevalence was 22% (29 cases in controls and 22 in the intervention group) with no significant difference between groups. At 6 weeks postpartum, controls had a [sim]3 fold greater weight retention, than the intervention group. A low intensity, behavioural lifestyle intervention, integrated with standard maternal care, is effective in optimising healthy gestational weight gain, improving physical activity in early pregnancy and in reducing postpartum weight retention. These results have significant public health implications.[br][br]Sources of Research Support: Australian New Zealand Clinical Trial Registry ACTRN12608000233325. This project is supported by a BRIDGES Grant from the Global Diabetes Foundation. BRIDGES, an International Diabetes Foundation project is supported by an educational grant from Eli Lilly and Company.[br][br]Nothing to Disclose: HJT, CL, CH 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3083 204 1738 SUN-LB6 PO02_LB Sunday 1535 2012


1532 ENDO12L_SUN-LB7 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) A Functional Growth Hormone Receptor Polymorphism, Exon 3 Deleted GHR, Is Associated with Radiographic Knee Osteoarthritis in Females with Familial Osteoarthritis at Multiple Sites: The GARP Study Kim Claessen, Margreet Kloppenburg, Herman Kroon, Jessica Bijsterbosch, Alberto Pereira, Hans Romijn, Tahar van der Straaten, Marjan Beekman, Eline Slagboom, Nienke Biermasz, Ingrid Meulenbelt Leiden University Medical Center, Leiden, Netherlands; Leiden University Medical Center, Leiden, Netherlands; Leiden University Medical Center, Leiden, Netherlands; Leiden University Medical Center, Leiden, Netherlands; Leiden University Medical Center, Leiden, Netherlands [bold][italic]Background[/italic][/bold][br]Acromegaly patients have increased GH and IGF1 levels from a GH-producing pituitary adenoma. One of the most invalidating long-term complications is arthropathy, with high disease activity as an important risk factor. Recently, presence of a growth hormone receptor (GHR) polymorphism, exon 3 deletion (d3-GHR), was reported to increase the risk of radiographic OA (ROA) in this patient group. This polymorphism, which results in increased GH sensitivity of the GHR, was also demonstrated to have functional consequences in various other clinical conditions, as illustrated by increased growth velocity after recombinant GH administration in children with growth failure.[br]Primary OA is a debilitating disease with a strong genetic component. Several lines of evidence indicate a role of the GH/IGF1 axis in primary OA onset, qualifying genetic variations in GH/IGF1 genes as obvious candidates for OA association studies.[br][bold][italic]Objective[/italic][/bold][br]To study the effects of the d3-GHR polymorphism on the extent and characteristics of ROA in patients with primary OA at multiple joint sites.[br][bold][italic]Methods[/italic][/bold][br]In a case-control study, we compared frequency of GHR[sub]fl-d3 [/sub]genotype between patients with familial primary OA from the GARP (Genetics, ARthrosis and Progression) Study, and controls. Kellgren-Lawrence scores were used to assess ROA in the knee, hip and hand; the Osteoarthritis Research Society atlas for the assessment of individual ROA features. Patients and controls were genotyped for 7 single nucleotide polymorphisms (SNPs) encompassing the d3-GHR gene to allow high throughput genotyping. One tagSNP was used as proxy for d3-GHR (full LD, pairwise r[sup]2[/sup]=1). Binary logistic regression analyses with robust standard errors were performed, to assess the relationship between d3-GHR and ROA.[br][bold][italic]Results[/italic][/bold][br]We studied 373 patients (mean age 60.1, 82% female) and 752 controls. GHR[sub]fl-d3 [/sub]genotype was significantly associated with ROA, especially in females (adjusted odds ratio (OR) (95%CI) 1.5(1.1-2.1), p=0.017). Strongest association was found with knee OA (adjusted OR 2.0(1.3-3.0), p=0.002), followed by hand OA (adjusted OR 1.5(1.1-2.1), p=0.024). No such a relationship was found in males. GHR[sub]fl-d3 [/sub]genotype was related to both osteophytes and joint space narrowing.[br][bold][italic]Conclusions[/italic][/bold][br]GHR[sub]d3-fl [/sub]genotype was associated with knee and hand ROA in females with a severe primary OA phenotype, indicating a role for the GH/IGF1 axis in the pathophysiology of primary OA. Further study is needed to confirm these results.[br][br]Nothing to Disclose: KC, MK, HK, JB, AP, HR, TvdS, MB, ES, NB, IM 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3089 204 1739 SUN-LB7 PO02_LB Sunday 1536 2012


1533 ENDO12L_SUN-LB8 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Modified-Release Formulations of Oral Hydrocortisone Create a Physiological Diurnal Cortisol Pattern [ndash] Novel Treatment Options for Congenital Adrenal Hyperplasia Miguel Debono, Martin Whitaker, Hiep Huatan, Cyrus Ghobadi, Geoff Tucker, Angela Taylor, John Newell-Price, Deborah Merke, Wiebke Arlt, Richard Ross University of Sheffield, Sheffield, South Yorkshire, UK; University of Sheffield, Sheffield, South Yorkshire, UK; 9, Betony Gardens, Kent, UK; Blades Enterprise Centre, Sheffield, UK; Blades Enterprise Centre, Sheffield, UK; University of Birmingham, Birmingham, UK; National Institutes of Health, Bethesda, MD Background: Cortisol has a distinct circadian rhythm. Current formulations of hydrocortisone are unable to replicate this rhythm so that patients with adrenal insufficiency wake with low cortisol levels and those with congenital adrenal hyperplasia high androgens. To address this problem we developed a modified release formulation of hydrocortisone, Chronocort[reg], to provide an overnight rise in cortisol (1). The initial formulation improved control of morning androgens in patients with CAH (2); however, the cortisol peak was too early and insufficiently sustained such that androgen levels rose during the day. The challenge we now address is the creation of a formulation delivered and absorbed within the limitations of the human gut transit time whilst generating a circadian cortisol profile that matches key physiological parameters.[br]Methods: We defined key parameters of the cortisol circadian rhythm through analysis of 24h cortisol profiles measured by LC-MS/MS in 28 normal volunteers. We then screened 6 Chronocort[reg] formulations with varying degrees of delay and sustained release to examine the impact on hydrocortisone pharmacokinetics. We tested formulations in a dog model and dexamethasone suppressed healthy volunteers (n=6 per group) and then compared them to key physiological parameters of the cortisol rhythm: Geometric mean for Cmax (mean of individual peaks), AUC (0[ndash]24hr) and Tmax.[br]Results: In normal individuals the Cmax was 21.5[micro]g/dl (594 nmol/l), AUC 170[micro]g/dl (4697 nmol/l.hr), Peak time 07:52. Release of hydrocortisone was achieved with a delay of 2-3 hours. Bioavailability was dependent on the degree of modified release. A 30mg formulation of Chronocort[reg] gave a Geomean (10th-90th percentile) Cmax of 25[micro]g/dl or 693nmol/l (22 [ndash] 29[micro]g/dl or 593 [ndash] 789nmol/l), AUC of 163[micro]g/dl.hr or 4502 nmol/l.hr (146 [ndash] 189[micro]g/dl.hr or 4021 [ndash] 5211nmol/l.hr), [gt]90% bioavailability and Tmax post-dosing of 8h (6h [ndash] 10h), within the range for physiological cortisol levels: Cmax 15[ndash]35[micro]g/dl (423 [ndash] 959 nmol/l), AUC 129 [ndash] 220 [micro]g/dl.hr (3560 [ndash] 6075nmol/l.hr), Peak time 05:54 [ndash] 09:06.[br]Conclusion: By adjusting the delayed and sustained release profile of modified release hydrocortisone we generated a formulation that can recreate a physiological Cmax, Tmax and AUC with [gt]90% bioavailability, thereby creating a uniquely suited tool for glucocorticoid replacement in adrenal insufficiency including congenital adrenal hyperplasia.[br][br](1) Debono et al., J Clin Endocrinol Metab. 2009 May; 94(5):1548-54[br](2) Verma et al., Clin Endocrinol (Oxf). 2010 Apr; 72(4):441-7.[br][br]Sources of Research Support: This research was funded by Diurnal Limited and supported in part by the Intramural Research Program of the NIH.[br][br] MW: Planning Group Member, Diurnal. HH: Consultant, Diurnal. GT: Director, OTHER, Diurnal. RR: Director, OTHER, Diurnal. Nothing to Disclose: MD, CG, AT, JN-P, DM, WA 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3009 204 1740 SUN-LB8 PO02_LB Sunday 1537 2012


1534 ENDO12L_SUN-LB9 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Elevated Serum Cortisol and Aldosterone during a Salt Losing Crisis Does Not Exclude Congenital Adrenal Hyperplasia Rashida Farhad Vasanwala, Ngee Lek, Fabian Yap KK Women[apos]s and Children[apos]s Hospital, Singapore, Singapore [bold]Background:[/bold] Children with salt wasting congenital adrenal hyperplasia (SW-CAH) cannot synthesize cortisol and aldosterone efficiently. If left untreated, the adrenal cortex is stimulated by ACTH and overproduces cortisol precursors and sex hormones. Aldosterone deficiency leads to salt wasting, failure to thrive and hypovolemic shock. We report 3 boys with SW-CAH who had uncharacteristically elevated cortisol and aldosterone levels at presentation.[br][bold]Clinical cases:[/bold] Presently 9 years, 8 years and 6 months old respectively, the boys presented with salt wasting crises at day 11, 12 and 60 of life and have since needed hydrocortisone and fludrocortisone replacement. Initial serum sodium levels were 120, 117 and 104 mmol/l; potassium levels were 9.3, 8.7 and 9 mmol/l; ACTH levels were 1398, 249.5 and 188.8 ng/l (normal 10-60) and 17OHP levels were 133.7, 4100 and 1165 nmol/l (normal [lt]6) respectively. Serum testosterone levels were [gt]55 nmol/l for the first two boys but was not done in the third. These clinical and biochemical features were in keeping with the diagnosis of SW-CAH.[br]Uncharacteristically, serum cortisol levels were 1820, 2810 and 5713 nmol/l (normal 46-626) and aldosterone levels were 813, [gt]3300 and [gt]4162 pmol/l (normal 93-834) respectively. All blood samples were collected prior to the administration of intravascular fluids, sodium and hydrocortisone. Cortisol was measured by sandwich immunoassay (Beckman, USA) and aldosterone was measured by radioimmunoassay (Zentech, Belgium).[br][bold]Conclusion:[/bold] While the diagnosis of CAH is established by elevated 17OHP, workup of an infant presenting with a salt losing crisis often includes measurement of serum cortisol and aldosterone levels. Our cases demonstrate that raised cortisol and aldosterone levels at presentation in children with SW-CAH does not necessarily exclude this diagnosis.[br][br]Nothing to Disclose: RFV, NL, FY 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3018 204 1741 SUN-LB9 PO02_LB Sunday 1538 2012


1535 ENDO12L_SUN-LB10 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Application of Stable Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry (LS-MS/MS) Methods for the Determination of the Androgen Metabolome in Serum in a Total Androgen Pathway Suppression Clinical Trial Daniel Tamae, Elahe A Mostaghel, Peter S Nelson, Paul Lange, Daniel Lin, Mary-Ellen Taplin, Steven Balk, William Ellis, Brett Marck, Robert Vessella, Bruce Montgomery, Trevor M Penning Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA; Fred Hutchinson Cancer Research Center, Seattle, WA; Fred Hutchinson Cancer Research Center, Seattle, WA; University of Washington, Seattle, WA; University of Washington, Seattle, WA; University of Washington, Seattle, WA; Harvard Medical School, Dana Farber/Harvard Cancer Center and Beth-Israel Deaconess Medical Center, Boston, MA; Education and Clinical Center, Veterans Affairs Puget Sound Health Care System, Seattle, WA Androgen deprivation therapy (ADT) is a cornerstone of prostate cancer treatment. However, after initial success with ADT, a subset of patients can develop a more aggressive, castration-resistant prostate cancer (CRPC). The vast majority of prostate cancer deaths arise from CRPC. The clinical efficacy of the CYP17A1 inhibitor, abiraterone acetate, in the treatment of CRPC confirms that the disease remains hormonally driven. Quantification of the androgen metabolome in serum and tumor biopsies can inform one as to the metabolic pathway that has been activated in CRPC tumors. This information can assist the clinician to custom tailor a therapeutic approach for maximum efficacy. To this end, a stable isotope dilution liquid chromatography electrospray ionization tandem mass spectrometry (SID-LC-ESI-MS/MS) method has been developed to detect ketosteroids as Girard T oximes and hydroxysteroids as picolinic esters. Accuracy and precision values have been obtained for ketosteroid quantification. Glucuronate and sulfate conjugates were quantitated using titered enzymatic hydrolysis. Limits of quantitation vary from metabolite to metabolite and range between 1-10 pg on column. We have applied our method to quantify serum androgen levels from patients enrolled in the Total Androgen Pathway Suppression (TAPS) Trial. In arm 1 of this trial, patients were given a combination of Goserelin and Bicalutamide; in arm 2, patients were given Goserelin and Dutasteride; in arm 3, patients were administered Goserelin, Dutasteride and Bicalutamide; and in the TAPS arm, arm 4, patients were given Goserelin, Dutasteride, Bicalutamide and Ketoconazole. In all four arms of the trial, we observed dramatic suppression of testosterone levels when comparing serum samples before and after treatment. In arms 2-4, we found statistically significant drops in androsterone and dihydrotestosterone levels. Finally, in the TAPS arm, we observed statistically significant suppression in DHEA-sulfate and DHEA-glucuronide levels in addition to all three of the aforementioned metabolites. This data was strongly concordant with quantification done using an alternative SID-LC-ESI-MS/MS assay conducted by an independent analyst.[br][br]Nothing to Disclose: DT, EAM, PSN, PL, DL, M-ET, SB, WE, BM, RV, BM, TMP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3019 204 1742 SUN-LB10 PO02_LB Sunday 1539 2012


1536 ENDO12L_SUN-LB11 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Racial/Ethnic Differences in Hepatic Steatosis in the Michigan Study of Women[apos]s Health across the Nation (SWAN) Catherine Kim, Siobhan Harlow, John Randolph, Margaret Helmuth, Ruth Carlos University of Michigan, Ann Arbor, MI; University of Michigan, Ann Arbor, MI; University of Michigan, Ann Arbor, MI; University of Michigan, Ann Arbor, MI Racial/ethnic differences in hepatic steatosis in a population-based cohort of mid-life women: the Michigan Study of Women[apos]s Health Across the Nation (SWAN)[br]Context: Hepatic steatosis is the most common liver disease in the U.S., due to its association with obesity and components of the metabolic syndrome. Ultrasound-diagnosed hepatic steatosis has also been associated with postmenopausal status in German and Asian populations. Reports conflict regarding differences in prevalence between African-American (AA) and Caucasian women. No studies have examined the association between ultrasound-diagnosed hepatic steatosis with sex steroid hormones [estradiol (E2) and testosterone (T)] or sex hormone binding globulin (SHBG).[br]Objectives: We examined the prevalence of ultrasound-diagnosed hepatic steatosis in postmenopausal Caucasian and AA women and associations with endogenous sex hormones in a population-based cohort.[br]Methods: SWAN is a longitudinal study designed to assess the impact of postmenopausal status on health. SWAN has annually assessed medical history, menstrual status, anthropometrics, blood pressure, sex hormone levels, insulin, glucose, and lipid levels. At the time of their year 13 exam, 345 (79% of the cohort) Michigan SWAN women opted to undergo ultrasound. Main outcome measures were the presence or absence of hepatic steatosis by race/ethnicity and associations with sex hormones after adjustment for known risk factors for steatosis. We also examined strength of risk factor associations by race/ethnicity.[br]Results: AAs (n=210) and Caucasians (n=135) were similar, except AAs had lower triglyceride levels and low-density lipoprotein cholesterol and higher systolic and diastolic blood pressure levels than Caucasians (p[lt]0.05 for all comparisons). Twenty-three percent of AAs had steatosis vs. 36% of Caucasians (p[lt]0.01). Risk factors for steatosis included greater age, waist circumference and lower high-density lipoprotein cholesterol and SHBG levels. After adjustment for these components, Caucasian race/ethnicity was still associated with greater odds of steatosis. AAs and Caucasians had similar risk factors for steatosis, except diabetic medication use was associated with lower odds of steatosis in Caucasians and not AAs.[br]Conclusions: Caucasians had greater odds of hepatic steatosis before and after adjustment for other risk factors. Lower SHBG levels were significantly associated with lower odds of steatosis.[br][br]Sources of Research Support: This work was supported by the National Institutes of Health (NIH), DHHS, through the National Institute on Aging (NIA), the National Institute of Nursing Research (NINR) and the NIH Office of Research on Women[apos]s Health (ORWH) (Grants NR004061; AG012505, AG012535, AG012531, AG012539, AG012546, AG012553, AG012554, AG012495). This work was also supported through the National Institute of Diabetes, Digestive, and Kidney Diseases (R01DK083297). The content of this article is solely the responsibility of the authors and does not necessarily represent the official views of the NIA, NINR, ORWH or the NIH. Additional contributions were made by the University of Michigan Center for Integrated Approaches to Complex Diseases, School of Public Health.[br][br]Nothing to Disclose: CK, SH, JR, MH, RC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3023 204 1743 SUN-LB11 PO02_LB Sunday 1540 2012


1537 ENDO12L_SUN-LB12 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Effects of Gastric Bypass Surgery on Female Reproductive Function Richard S Legro, William C Dodson, Carol L Gnatuk, Stephanie J Estes, Allen R Kunselman, Juliana W Meadows, James S Kesner, Edward F Krieg, Jr, Ann M Rogers, Randy S Haluck, Robert N Cooney Pennsylvania State University College of Medicine, Hershey, PA; Pennsylvania State University College of Medicine, Hershey, PA; National Institute for Occupational Safety and Health, Cincinnati, OH; Pennsylvania State University College of Medicine, Hershey, PA; SUNY Upstate Medical Center, Syracuse, NY Obesity is associated with reproductive abnormalities and decreased fecundity in women. Improvements in reproductive function have been noted after bariatric surgery, though the mechanisms and time-related changes are not fully understood.[br][bold]Objective [/bold]To determine whether women experience an improved frequency or quality of ovulation with associated reproductive changes following Roux en Y gastric bypass surgery.[br][bold]Design [/bold]A prospective cohort study that enrolled female subjects from 2005 to 2008 with study visits one month prior, one month after, and then at 3,6, 12, and up to 24 months after surgery.[br][bold]Setting: [/bold]Penn State Hershey Medical Center.[br][bold]Patients [/bold]29 obese women of reproductive age not using confounding medications.[br][bold]Main Outcome Measures [/bold]Primary outcome: Integrated levels of urinary progestin (pregnanediol 3-glururonide; Pd3G) from daily urinary collections at 12 months post-operatively as an indication of the frequency and quality of ovulation. Secondary outcomes: vaginal bleeding, other endocrine (serum [amp] urinary) measures of menstrual cycle function, pelvic ultrasound, changes in body composition by DXA, and female sexual function index.[br][bold]Results[/bold] There was no change in the frequency of ovulation ([ge] 90% at all timepoints) or luteal Pd3G levels over the course of the study. The follicular phase was 6.5 days shorter within 3 months after surgery, and 7.9-8.9 days shorter 6-24 months post surgery (P [lt] 0.01). This effect was also manifested in the reduction of three other menstrual cycle function parameters: days from E13G rise onset to ovulation (P=0.012), day of Pd3G peak (P=0.002), and suggested by day of E13G peak (P=0.06) Biochemical hyperandrogenism improved largely due to an immediate post operative increase in serum sex hormone binding globulin levels (P [lt] 0.01), with no effects over time on parameters of clinical hyperandrogenism (sebum production, acne, hirsutism). Bone density was preserved, contrasting with significant loss of lean muscle mass (P [lt] 0.001) and fat (P [lt] 0.001), reflecting preferential abdominal fat loss (P [lt] 0.001). There were no changes in ovarian volume or largest follicle size over time. Female sexual function had improved 28% (P=0.02) by 12 months.[br][bold]Conclusions[/bold] Ovulation persists in spite of morbid obesity, acutely decreased caloric intake, and massive weight loss after bypass surgery. Improved fecundity after surgery may be related to shortening of the follicular phase and/or by improved female sexual function.[br][br]Sources of Research Support: Tobacco Settlement Funds from the Commonwealth of Pennsylvania; Clinical Translational Science Award Grant, M01RR10732 and Construction Grant to Pennsylvania State University.[br][br] SJE: Proctor, OTHER, Intuitive Surgical (da vinci), Proctor, Hologic (myosure). Nothing to Disclose: RSL, WCD, CLG, ARK, JWM, JSK, EFK, Jr, AMR, RSH, RNC 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3025 204 1744 SUN-LB12 PO02_LB Sunday 1541 2012


1538 ENDO12L_SUN-LB13 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Dose Sparing and Safety-Enhancing Effects of an IGF-I-Based Dosing Regimen in Short Children Treated with Growth Hormone in a 2-Year Randomized Controlled Trial: Therapeutic and Pharmacoeconomic Considerations Pinchas Cohen, Alan D Rogol, Wayne Weng, Anne-Marie Kappelgaard, Ron G Rosenfeld, John Germak Mattel Children[apos]s Hospital at UCLA, Los Angeles, CA; University of Virginia, Charlottesville, VA; Novo Nordisk Inc, Princeton, NJ; Novo Nordisk A/S, S[oslash]borg, Denmark; Oregon Health [amp] Science University, Portland, OR [bold]BACKGROUND: [/bold]A number of approaches have been explored to optimize the efficacy and safety of growth hormone therapy (GHT) in children with short stature, including IGF-I targeted GH dosing. In addition to the clinical outcomes, it is important to evaluate such treatment regimens from cost effectiveness and theoretical safety perspectives.[br][bold]OBJECTIVE:[/bold] To assess cost effectiveness (as measured by change in height standard deviation score ([Delta]HSDS) per mg/kg GH dose) and theoretical safety (as measured by the proportion of IGF-I measurements above +2 SDS) of GHT at the end of a 2-year randomized, controlled trial. [br][bold]DESIGN/METHODS:[/bold] 172 prepubertal short children (age 7.5[plusmn]2.4 y, HSDS -2.64[plusmn]0.61) with low IGF-I levels were randomized to 1 of 3 GHT groups: conventional weight-based dosing (0.04 mg/kg/d) (Conv), or to GH dosing titrated to an IGF-I target of 0[plusmn]0.5 SDS (RCC1) or to 2[plusmn]0.5 SDS (RCC2). Patients were classified as GH deficiency (GHD) (peak GH[lt]7 ng/mL) or idiopathic short stature (ISS) ([ge]7 ng/mL) based on GH stimulation testing.[br][bold]RESULTS:[/bold] In ISS patients at Year 2, mean [Delta]HSDS and GH dose (mg/kg/day) were 0.87, 0.04 for Conv; 0.84, 0.032 for RCC1; 1.33, 0.114 for RCC2 respectively. In GHD patients the values were 1.23, 0.04 for Conv; 1.41, 0.037 for RCC1; 2.04, 0.091 for RCC2 respectively. The cost-effectiveness measure of [Delta]HSDS/GH-dose ratio was higher for RCC1 than Conv in both ISS (32.5 vs. 21.3, p=0.005) and GHD (48.1 vs. 30.3, p=0.02) groups. Although the RCC2 group achieved greater [Delta]HSDS compared to Conv and RCC1 groups, the GH dose was also significantly higher than the other two groups and the [Delta]HSDS/GH dose ratio for RCC2 (16.3 for ISS and 32.7 for GHD) was lower than RCC1 and not different from Conv. In comparison to ISS patients using the same regimen, GHD patients showed a greater [Delta]HSDS/GH dose ratio. While mean IGF-I SDS levels were not different between the RCC1 and Conv groups (0.23 vs 0.85, p=0.079), IGF-I SDS levels [gt] +2 SDS at Year 2 were observed in 7% of patients in RCC1 compared to 30% in Conv (p[lt]0.01).[br][bold]CONCLUSIONS: [/bold]IGF-I targeted GH dosing to the age- and gender-adjusted mean (zero-SDS) resulted in higher [Delta]HSDS/GH-dose ratio than conventional weight-based dosing with comparable levels of IGF-I and [Delta]HSDS achieved in both GHD and ISS patients. Therefore, compared to conventional dosing, IGF-based dosing targeted to the mean SDS was associated with avoidance of exposure to higher IGF-I levels, as well as a more cost effective mode of GHT.[br][br]PC: Consultant, Novo Nordisk. ADR: Consultant, Novo Nordisk. WW: Employee, Novo Nordisk. A-MK: Employee, Novo Nordisk. JG: Employee, Novo Nordisk. Nothing to Disclose: , RGR 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3026 204 1745 SUN-LB13 PO02_LB Sunday 1542 2012


1539 ENDO12L_SUN-LB14 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Complete Reversal of Severe Osteoporosis Due to Osteomalacia from Tenofovir-Induced Fanconi[apos]s Syndrome Luis Anaya, Kavitha Beedupalli, Jan M Bruder UTHSCSA, San Antonio, TX [bold]Background: [/bold]Tenofovir use as part of HAART therapy for HIV is associated with a decrease in bone mineral density (BMD), proximal renal tubule dysfunction and Fanconi[apos]s syndrome. However, the clinical implication of this is not clear and studies do not show increased incidence of fracture. [bold]Case: [/bold]48 y/o Caucasian man with HIV and HBV infection on Tenofovir, was admitted with right hip fracture after a fall from standing height. He reported months of generalized bone pain and marked proximal muscle weakness requiring a cane for ambulation. Admission labs revealed hypokalemia (2.8 mmol/L), hypophosphatemia (1.3 mg/dL), metabolic acidosis and an elevated alkaline phosphatase level (444 IU/L). Further workup revealed significant proteinuria, aminoaciduria, glucosuria and renal phosphate wasting (514 mg/day); inappropriately low 1,25-dihydroxyvitamin D[sub]3 [/sub](9 pg/ml), and low 25(OH) Vit D (14ng/ml) levels. Serum calcium, intact PTH and FGF-23 levels were within normal limits. BMD by dual-energy x-ray absorptiometry (DXA) at all sites were low (L1-L4 0.705gr/cm[sup]2[/sup], T-score -4.3, Z-score -3.4; and left total hip 0.475gr/cm[sup]2[/sup], T-score -4.3, Z-score -3.5). The constellation of these biochemical abnormalities suggested osteomalacia with phosphate wasting due to Tenofovir-induced Fanconi[apos]s syndrome. Tenofovir was discontinued. Phosphate supplements and Rocaltrol (0.5mcg/d) and weekly ergocalciferol were begun. His symptoms improved markedly within four months after stopping the Tenofovir. He was able to walk without support and the phosphate supplements were discontinued. Electrolytes and phosphate level return to normal with improvement in alkaline phosphatase level. Repeat BMD by DXA a year later showed a significant increase of BMD of greater than 100% at the spine with complete normalization. Significant improvement was also seen at the hip (L1-L4, 1.421gr/cm[sup]2[/sup], T-score 1.7, Z-score 2.4; and left hip 0.748gr/cm2, T-score -2.4, Z-score -1.8). [bold]Conclusion:[/bold] To our knowledge this is the first case of severe osteoporosis diagnosed by BMD and hip fracture due to osteomalacia from Tenofovir-induced Fanconi[apos]s syndrome with complete resolution after a year of stopping Tenfovir. Since Tenofovir is frequently used in HAART therapy, patients should be monitored for osteoporosis and Fanconi[apos]s syndrome.[br][br]Nothing to Disclose: LA, KB, JMB 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3043 204 1746 SUN-LB14 PO02_LB Sunday 1543 2012


1540 ENDO12L_SUN-LB15 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Utility and Limit of Adrenocorticotropin Loading Test in Discriminating Aldosterone Producing Adenoma [ndash] Analysis of 256 Patients with Primary Aldosteronism and 206 Patients with Primary Hypertension Ryo Morimoto, Masataka Kudo, Yoshitsugu Iwakura, Yoshikiyo Ono, Kei Takase, Hironobu Sasano, Sadayoshi Ito, Fumitoshi Satoh Tohoku University Hospital, Sendai, Japan; Tohoku University Hospital, Sendai, Japan; Tohoku University Hospital, Sendai, Japan Background: Adrenal vein sampling (AVS) is the most accurate diagnostic tool in differentiating aldosterone producing adenomas (APA) from bilateral hyperaldosteronism (BHA) but needs specialized and interventional techniques. APA is generally reported to be more responsive to ACTH than BHA. Objective: The aim was to evaluate the diagnostic value of ACTH loading test in discriminating APA from BHA or primary hypertension (PH). Methods: The all patients with APA and BHA diagnosed by more than 20 of aldosterone/renin activity ratio (ng/dl per ng/ml/hr) under captopril challenging test (CCT), and underwent AVS. Adenomas were surgically proven and confirmed by histological diagnosis including immunehistochemical analysis of steroidogenic enzymes in all APA patients. ACTH loading tests were performed in 119 APA, 135 BHA and 204 PH patients, plasma aldosterone concentrations (PAC) were measured at 30, 60 and 90 min after ACTH loading.[br]Results: The peak values of PAC after ACTH loading in the patients with APA (51.8 ng/dl) were significantly higher than in those with BHA (28.6 ng/dl) or PH (27.6 ng/dl). ROC curve analysis of the peak PAC after ACTH loading in APA patients versus BHA or PH patients demonstrated that APA was diagnosed by the optimal cut-off value above 36.4 ng/dl with sensitivity and specificity of 68.8% and 83.8% respectively. The analysis of the peak PAC in APA patients versus PH patients with PRA below 1.0 ng/ml/hr indicated that APA was diagnosed by the cut-off value above 31.3 ng/dl with sensitivity and specificity of 80.7% and 85.6% respectively. Conclusions: Our study in 254 patients with primary aldosteronism showed that ACTH loading test might be tentatively useful, and its sensitivity would be lower than that of the previous report in smaller numbers of patients (1). Nevertheless, the test could increase its accuracy in discriminating APA patients from low renin PH patients.[br][br](1) Sonoyama T et al., J Clin Endocrinol Metab. 2011; 96(9):2771.[br][br]Nothing to Disclose: RM, MK, YI, YO, KT, HS, SI, FS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3048 204 1747 SUN-LB15 PO02_LB Sunday 1544 2012


1541 ENDO12L_SUN-LB16 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Metformin Therapy and Assessment for Vitamin B [sub]12[/sub] Deficiency: Is It Necessary? Sheharyar Qureshi, Peter Winocour West Middlesex University Hospital, London, UK; QE2 Hospital, East and North Hertfordshire NHS Trust, Hertfordshire, UK Metformin therapy in type 2 diabetes mellitus (T2DM) has been recognised as a cause of vitamin B[sub]12[/sub] deficiency for at least 40 years, but routine measurement is not currently advocated in clinical guidelines. Assessment might be of particular relevance in T2DM complicated by peripheral neuropathy.[br]This service review examined whether serum vitamin B[sub]12[/sub] levels were measured in patients with high dose ([gt]2g/day) and long term (four years) Metformin treatment, in particular among those with peripheral neuropathy. We also evaluated the effectiveness of vitamin B[sub]12[/sub] replacement when levels were low.[br]Of 283 patients on high dose Metformin for more than four years only, 70 (25%) had vitamin B[sub]12[/sub] levels checked. All of these 70 cases had peripheral neuropathy. Vitamin B[sub]12[/sub] deficiency ([lt]150pg/ml) was recorded in 23 (33%). Where vitamin B[sub]12[/sub] levels were deficient, replacement vitamin B[sub]12[/sub] was documented in only two (2.9%) patients and improvement in neuropathic symptoms post treatment were documented in only four (5.7%) patients.[br]Conclusion: Vitamin B[sub]12[/sub] levels were measured infrequently in T2DM, in particular among those with peripheral neuropathy. Levels were frequently low when assessed among T2DM patients with peripheral neuropathy. A record that vitamin B[sub]12[/sub] therapy was initiated was only made in a small number of cases, so the impact on peripheral neuropathy was unclear.[br]Recommendations: All patients with T2DM on long-term treatment with high dose Metformin should be assessed for vitamin B[sub]12[/sub] deficiency, particularly if complicated by peripheral neuropathy, and then considered for parenteral vitamin B[sub]12[/sub] replacement if deficient.[br][br](1) Kirpichnikov D, et al. Metformin: an update. Ann Intern Med 2002;137:25[ndash]33.[br](2) UK Prospective Diabetes Study (UKPDS) Group.Effect of intensive blood-glucose control with metformin on complications in overweight patients with type 2 diabetes (UKPDS 34). Lancet 1998;352:854[ndash]65.[br](3) Derosa G, et al. Comparison of glycaemic control and cardiovascular risk profile in patients with type 2 diabetes during treatment with either repaglinide or metformin. Diabetes Res Clin Pract 2003;60:161[ndash]9.[br](4) Tomkin GH, et al. Vitamin B12 status of patients on long-term metformin therapy. BMJ 1971;2:685[ndash]7.[br](5)Jager D, et al. Long term treatment with metformin in patients with type 2 diabetes and risk of vitamin B12 deficiency: randomised placebo controlled trial. BMJ 2010;340:c2181.[br](6) Andr[egrave]s E, et al. Vitamin B12 (cobalamin) deficiency[br]in elderly patients. CMAJ 2004;171:251[ndash]9.[br](7) Adams JF, et al. Malabsorption of vitamin B12 and intrinsic factor secretion during biguanide therapy.Diabetologia 1983;24:16[ndash]8.[br](8) Ting RZ-W, et al. Risk factors of vitamin B12 deficiency[br]in patients receiving metformin. Arch Intern Med 2006;166:1975[ndash]9.[br](9) DeFronzo RA, et al. The Multicenter Metformin Study Group. Efficacy of metformin in patients with non-insulin-dependent diabetes mellitus. N Engl J Med 1995;333:541[ndash]9.[br](10) Bauman WA, et al. Increased intake of calcium reverses vitamin B12 malabsorption induced by metformin. Diabetes Care 2000;23:1227[ndash]31.[br](11) Wulffel[eacute] MG, et al. Effects of short-term treatment with metformin on serum concentrations of homocysteine,folate, and vitamin B12 in type 2 diabetes mellitus: a randomized placebo trial. J Intern Med 2003;254:455[ndash]63.[br](12) Ban-Hok, et al. Pernicious anaemia. N Engl J Med 1997;337:1441[ndash]8[br](13) Lindenbaum J, et al. Neuropsychiatric disorders[br]cased by cobalamin deficiency in the absence of anaemia and macrocytosis. N Eng J Med 1988;318:1720[ndash]8.[br](14) Fine EJ, et al. The neurophysiological profile of B12[br]deficiency. Muscle Nerve 1990;13:158[ndash]64.[br](15) Young MJ, et al. A multicentre study of the prevalence[br]of diabetic peripheral neuropathy in the UK hospital clinic population. Diabetologia 1993;36:150[ndash]6.[br](16) Oral or intramuscular vitamin B12? (Relevant BNF section: 9.1.2). Drug Ther Bull 2009;47:19[ndash]21 doi:10.1136/dtb.2009.01.0002[br](17) Joint Formulary Committee. British National Formulary, edn 56. BMJ Publishing Group and Royal Pharmaceutical Society of Great Britain, 2008.[br](18) Kuzminski AM, et al. Effective treatment of cobalamin[br]deficiency with oral cobalamin. Blood 1998;92(4):1191[ndash]8.[br][br]Sources of Research Support: Novo Nordisk Ltd for registration for the meeting.[br][br]Nothing to Disclose: SQ, PW 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3055 204 1748 SUN-LB16 PO02_LB Sunday 1545 2012


1542 ENDO12L_SUN-LB17 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Retrospective Review of the Ultrasound Characteristics of Thyroid Nodule and Pathology Results in Caucasian Versus African American Patients Margaret Gladysz, Dale Kraemer, Mae Sheikh-Ali University of Florida, Jacksonville, FL [bold]Objectives: [/bold][br]A rational approach to evaluate and manage thyroid nodules is based on the clinician[apos]s ability to distinguish the benign from malignant nodules in a highly reliable and cost-effective manner. African-American race and increasing age were found to be independent risk factors for the development of large nodular goiter. Yet the differences in the characteristics of the thyroid nodules and its correlation with fine needle aspiration and /or surgical pathology report in African American versus Caucasians have not been examined.[br][bold]Methods: [/bold][br]We performed retrospective, observational study on patients with diagnosis of multinodular or uninodular goiter that were seen at University of Florida, Jacksonville endocrinology clinics between January 1, 2010 and February 29, 2012.[br]Variables examined included the patient[apos]s ethnic origin, sex and age at presentation, nodule size, internal content of the nodule, its margins, echogenicity of solid portions, the presence and type of calcifications, vascularity, and presence or absence of the abnormal lymph nodes.[br][bold]Results: [/bold][br]Out of 290 identified patients, 170 were included with total of 214 nodules. 94.22% (n=163) patients were females. Mean age of patients was 54.3 years old (range: 27-80). The mean size of the studied nodules was 2.48cm (SD [plusmn]1.14, nodule size range: 0.80 to 7.7cm in the biggest dimension). 65% (n=128) of nodules were found in African American population and 35% (n=69) were in Caucasian population.[br]Out of 214 nodules, 4.7% (n=10) were malignant, and 95.3% (n=204) were found to be benign.[br]Of the 128 nodules that were identified in the African American population from which 5.47% (n=7) were malignant and 94.53% (n=121) were benign. In Caucasian population, total of 71 nodules were identified. 2.9% were found to be malignant (n=2), and 97.1% (n=69) were benign.[br]There was no statistical significance in the malignancy comparing both ethnic groups (P value is 0.4).[br]In addition there was no statistical significance when comparing the characteristic of thyroid nodules in African American and Caucasian: (P value for all 0.1-0.7).[br][bold]Conclusion: [/bold][br]1. No statistically significant difference was found in the malignancy incidence and thyroid nodule characteristics based on the race (African American versus Caucasian) in our patient population.[br]2. In assessment of the US findings, we found that there was no evaluation of the lymph nodes which is a very important aspect in evaluating the risk of malignancy.[br][br]1. An investigation of epidemiologic factors associated with large nodular goiter. Phitayakorn R, Super DM, McHenry CR. J Surg Res. 2006 Jun 1;133(1):16-21.[br]2. Thyroid oncology in everyday practice. Apropos of 105 cases. Ndiaye I, Diouf R, Diop EM, Ndiaye PD. Ann Otolaryngol Chir Cervicofac. 1993;110(4):211-7.[br]3. Thyroid cancer: is ethnicity relevant? T Galm, S Minhas, RJ Cullen, H Griffiths. The Journal of Laryngology [amp] Otology (2011), 125, 816[ndash]819.[br]4. L. Hegedus. Clinical practice. The thyroid nodule. N Engl J Med 2004: 351:1764[ndash]1771.[br]5. Moon WJ, Jung SL, Lee JH, et al. Benign and malignant thyroid nodules: US differentiation[mdash]multicenter retrospective study. Radiology 2008; 247:762[ndash]770[br]6. Su-kyoung Jeh, So Lyung Jung, Bum Soo Kim, Yoen Soo Lee. Evaluating the Degree of Conformity of Papillary Carcinoma and Follicular Carcinoma to the Reported Ultrasonographic Findings of Malignant Thyroid Tumor. Korean J Radiol 2007;8:192-197.[br]7. Zhang YX, et. al. Fine-needle aspiration cytology of thyroid nodules: a clinical evaluation. 2011 Nov;46(11):892-6.[br]8. Papini E, Guglielmi R, Bianchini A, et al. Risk of malignancy in nonpalpable thyroid nodules: predictive value of ultrasound and color-Doppler features. J Clin Endocrinol Metab 2002; 87:1941[ndash]1946. Kim E, Park CS, Chung WY, et al.[br][br]Nothing to Disclose: MG, DK, MS-A 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3063 204 1749 SUN-LB17 PO02_LB Sunday 1546 2012


1543 ENDO12L_SUN-LB18 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Short-Term Testosterone Treatment Suppresses Skeletal Muscle NIK Expression in Older Men Astrid M Horstman, Sanjeev Choudhary, William J Durham, E Lichar Dillon, Ronald G Tilton, Randall J Urban, Melinda Sheffield-Moore University of Texas Medical Branch, Galveston, TX A number of events are known to trigger muscle loss with aging including inflammation, inactivity, and loss of anabolic hormones such as testosterone. It is unclear, however, what role NF-[kappa]B inducing kinase (NIK), an upstream regulatory MAP kinase in the NF-[kappa]B activation pathway, plays in mediating age-related muscle loss. Our data show that a link exists between increased NIK and decreased Notch levels in aging skeletal muscle. Moreover, testosterone has been shown to exert anti-catabolic properties in the skeletal muscle of older men. Thus, we hypothesized that testosterone may be involved in regulating NIK levels, as NIK has been shown to be significantly elevated in aging and diseases where chronic inflammation is present. We investigated whether short-term administration (7 days) of testosterone was capable of counteracting aberrantly regulated NIK signaling in older men with serum testosterone concentrations in the low normal range ([le]500 ng/dl), suggestive of a possible NIK-testosterone link in counteracting muscle loss with aging. Blood was collected and biopsies were obtained from the mixed [italic]vastus lateralis[/italic] muscle before and 7 days after either a single dose (intramuscular injection) of testosterone enanthate (100 mg on Day 1) or initiation of daily topical testosterone gel (two 5g packets of 1%AndroGel[reg]/day). Muscle samples were homogenized and protein lysates were fractionated on 10% SDS-PAGE and immunoblotted for NIK using specific antibodies. GAPDH served as a loading control. Serum testosterone was measured on an Immulite 2000. Skeletal muscle NIK levels significantly decreased following 7 days of testosterone treatment. Further, we found a significant negative association (r=-0.841, p[lt]0.036) between baseline serum testosterone and skeletal muscle NIK protein levels. Moreover, a negative correlation was found between the [italic]change[/italic] in serum testosterone levels and the [italic]change[/italic] in NIK protein expression after one- week treatment (r=-0.412), although this correlation did not reach statistical significance due to the small sample size. These data clearly demonstrate that as little as seven days of testosterone treatment can suppress skeletal muscle NIK levels in older men, suggesting that testosterone may exert its anti-catabolic properties in skeletal muscle via its regulation of NIK.[br][br]Sources of Research Support: The Moody Endowment.[br][br]Nothing to Disclose: AMH, SC, WJD, ELD, RGT, RJU, MS-M 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3071 204 1750 SUN-LB18 PO02_LB Sunday 1547 2012


1544 ENDO12L_SUN-LB19 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Glycemic Variation in Non-Diabetic Morbidly Obese Applicants for The Biggest Loser Television Show Sara J Salkind, Robert Huizenga, Stephanie J Fonda, Robert A Vigersky Walter Reed National Military Medical Center, Bethesda, MD; University of California, Los Angeles, CA Despite the controversy about what role post-prandial hyperglycemia and overall glycemic variation play in the development of micro- and macrovascular disease in Type 2 diabetes, there is a scarcity of data about glycemic variability in the obese and morbidly obese individuals without diabetes - a population at increased risk for cardiovascular disease and diabetes. Therefore, we studied the glycemic variability of morbidly obese applicants to The Biggest Loser television show. Twenty-two of 43 morbidly obese applicants who had no evidence of diabetes - A1C [lt]5.7%, fasting glucose [lt]100 mg/dl, and 2-hour post-glucose load [lt]140 mg/dl) - wore a masked DexCom Seven Plus[reg] continuous glucose monitor (CGM) for 5-7 days. There were 12 Caucasians, 5 African-Americans, 2 Hispanics, and 3 [ldquo]others[rdquo]; 14 were women and 8 were men. Their mean (SD) age was 32 ([plusmn]10.4) years, BMI was 49.64 ([plusmn]10.1), and DXA-derived % body fat was 57 ([plusmn]5) %. Sixteen individuals had hypertension. Their CBC, renal, and hepatic function were normal. LDL was 119 mg/dl ([plusmn]29) and triglycerides were102 mg/dl ([plusmn]50). These individuals had a mean ([plusmn]SD) A1C of 5.3% ([plusmn]0.2), fasting plasma glucose of 89.6 mg/dl ([plusmn]12.5), and a 2-hour post-glucose load of 106.7 ([plusmn]18.5) mg/dl. Their overall glycemic variability (SD) for 5-7 days measure by CGM was 19.1 (range 8.8 [ndash] 30) and a coefficient of variation of 18.8%. This glycemic variability is approximately 50% greater than that seen in 20 similarly aged, non-diabetic individuals whose A1C and overall CGM[ndash]derived glucose variability was 5.2 ([plusmn]0.3%) and 12.6 mg/dl, respectively (1). Only 0.3% of values were [gt]140 mg/dl in the JDRF group whereas 27.3% were [gt]140 mg/dl in the morbidly obese population. Nineteen of the 22 contestants had measurement of carotid intima media thickness. It was increased in 7 contestants (0.74[plusmn]0.07) and normal in 12 (0.56[plusmn]0.05 mm). These data suggest that morbidly obese applicants to The Biggest Loser television program have increased glycemic variability which may put them at greater risk of cardiovascular disease due to this dysglycemia-induced oxidative stress. The data also suggest that it may be possible to use CGM to educate morbidly obese people on the effects of their lifestyle choices on the importance of glycemic variability.[br][br](1) Juvenile Diabetes Research Foundation Continuous GLucose Monitoring Study Group, Diabetes Care 2010; 33:1297.[br][br]Sources of Research Support: DexCom, Inc.[br][br]Nothing to Disclose: SJS, RH, SJF, RAV 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3075 204 1751 SUN-LB19 PO02_LB Sunday 1548 2012


1545 ENDO12L_SUN-LB20 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Insulin Delivery Using Hollow Microneedles in Children with Type 1 Diabetes Eric Ian Felner, James Jefferis Norman, Nicholas A Raviele, Milton R Brown, Mark R Prausnitz Emory University School of Medicine, Atlanta, GA; Georgia Institute of Technology, Atlanta, GA [bold]Introduction:[/bold] Treatment of Type 1 diabetes (T1D) is limited by inadequate methods of insulin delivery that require painful injections and frequent glucose monitoring.(1) Children in particular often omit insulin injections and glucose monitoring due to the pain and apprehension associated with needles.(2) Microneedles (MNs) may provide a better interface to deliver insulin because of painless administration. Due to rapid insulin pharmacokinetics,(3) MNs may also reduce the need for glucose monitoring by enabling automatic closed-loop insulin therapy. We conducted the first study of MN-based insulin delivery in children with T1D to examine pain and pharmacokinetics compared to a subcutaneous infusion catheter (SIC).[br][bold]Methods:[/bold] Hollow glass MNs were fabricated and utilized according to Gupta et al.(3) Sixteen subjects (10-17 yr) with T1D for [ge] 2-yr who were using insulin pump therapy for [ge] 1 yr were enrolled. Subjects received both delivery methods on separate days. The order of methods was randomized. Immediately prior to delivery of Lispro insulin, a baseline blood sample was collected. Immediately after insulin delivery, a 75-gram carbohydrate meal was eaten. Blood was sampled every 15 min for 2 hr and then every 30 min for 2 hr. Subjects rated the pain of insertion (PI) and pain with insulin delivery (PD) for both methods using a visual analog pain scale (VAS). We compared PI, PD, time to peak insulin concentration (t-max), peak insulin concentration (C-max), area-under-insulin curve (AUIC), and area-under-glucose curve (AUGC) with paired statistical analyses. A compartment model was fit to the insulin data using nonlinear regression.[br][bold]Results: [/bold]Three subjects were excluded because of incomplete delivery with the MN and one was excluded due to confounding insulin delivery before the experiment. MN insertion resulted in less PI compared to SIC insertion (p = 0.01). No significant difference was observed in PD. The compartment model showed a 3.7-fold increase in absorption coefficient for MN delivery (p = 0.0002) but no significant difference in bioavailability (p = 0.21), elimination coefficient (p = 0.11), or C-max (p = 0.57). As a result of increased absorption, t-max was significantly reduced (p = 0.0004).[br][bold]Conclusions: [/bold]Delivery of Lispro insulin through a MN was less painful and had a more rapid onset of action as compared to a SIC. This is a promising result for improving the compliance of insulin delivery in children with T1D.[br][br]1. Mollema ED et al.,Diabetic Med 2001; 18(8):671-4.[br]2. Hanas R,Pedi Diabetes 2004; 5(2):102-111.[br]3. Gupta J et al.,Diabetes Technol Ther 2011; 13(4):451-6.[br][br]Sources of Research Support: Thrasher Research Fund 02827-8 awarded to EIF.[br][br]Nothing to Disclose: EIF, JJN, NAR, MRB, MRP 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3080 204 1752 SUN-LB20 PO02_LB Sunday 1549 2012


1546 ENDO12L_SUN-LB21 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) Risk Factors of Renal Damage and Prevalence of CKD in Primary Aldosteronism Yoshitsugu Iwakura, Ryo Morimoto, Yoshikiyo Ono, Masataka Kudo, Kei Takase, Hironobu Sasano, Sadayoshi Ito, Fumitoshi Satoh Tohoku University Hospital, Sendai, Japan; Tohoku University Hospital, Sendai, Japan; Tohoku University Hospital, Sendai, Japan Purpose: Higher prevalence of urinary albumin was reported in patients with primary aldosteronism (PA) than those with essential hypertension (1). Although glomerular hyperfiltration was hypothesized as one of mechanisms of urinary albumin and this condition was considered to overestimate true renal function and mask real renal damage, it remains to be unclear. This prospective study was designed to identify risk factors of urinary albumin and to clarify prevalence of chronic kidney disease (CKD) in PA.[br]Methods: 213 patients were diagnosed and distributed according to the results of adrenal venous sampling. 102 patients with aldosterone producing adenoma underwent adrenalectomy and 111 were treated with mineralocorticoid receptor antagonists. Blood pressure (BP), urinary albumin excretion (UAE) and estimated glomerular filtration rate (eGFR) were followed up during 12-month period after treatments. CKD was defined as less than 60 ml/min/1.73m2 in eGFR.[br]Results: BP, UAE, and eGFR were significantly reduced at 1 month after treatment as compared with those at the first visit and these remained stable throughout the follow-up period of 12 months. Prevalence of CKD increased approximately 20% after treatments. Multivariate regression analysis showed that plasma aldosterone and BP were strong independent risk factors of higher albuminuria before intervention, and UAE was a predictor of decrement of eGFR after treatments.[br]Conclusions: Based on the present findings, early diagnosis and treatment of PA should be recommended in those with albuminuria to prevent renal damage associated with high levels of aldosterone.[br][br](1) Ribstein J et al. J Am Soc Nephrol. 2005;16:1320.[br][br]Nothing to Disclose: YI, RM, YO, MK, KT, HS, SI, FS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3085 204 1753 SUN-LB21 PO02_LB Sunday 1550 2012


1547 ENDO12L_SUN-LB22 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) The Effect of Controlled Aerobic Exercise on Endothelial Dysfunction in Patients with Pre-Diabetes: A Crossover Pilot Study Pragathi Saligram, Mary Young, Ann Lagoy, Cyril Chou, Sarah Witkowski, Ashequl Islam, Sabyasachi Sen Baystate Medical Center/Tufts University, Springfield, MA; Baystate Medical Center/Tufts University, Springfield, MA; University of Massachusetts, Amherst, MA Introduction: At least 57 million US adults have pre-diabetes. Pre-diabetes has the potential to cause endothelial dysfunction by altering function and gene expression of endothelial progenitor cells as well as mature endothelial cells. We hypothesize that aerobic exercise will reduce endothelial inflammation and improve function of endothelial progenitor cells (EPC) in pre-diabetes. Though life-style modification has been shown to prevent progression from Pre-diabetes to overt diabetes, this hypothesis has not been tested.[br]Methods: This is a crossover study of 16-week duration, using exercise-naive pre-diabetes patients, aged 45-65 yrs with a BMI of 25-34.9. We studied their vascular reactivity by flow mediated dilatation (FMD), EPC function and gene expression and serum endothelial inflammatory levels after 6 weeks each of aerobic exercise (150min/week) and non exercise phase, in a cross over design with 4 week wash-out period between the 2 phases. The functional assays of EPCs noted were migration in response to chemotactic factors such as Vascular Endothelial Growth Factor (VEGF-A, 0- 50ng/ml) and Stromal Cell-Derived Factor-1(SDF1[alpha], 0-100 ng/ml) and tube formation assays. Adherence to exercise regimen was confirmed by regular phone calls and downloadable accelerometers.[br]Results: FMD studies (undertaken by 3 observers) showed mean FMD in non-exercise group of 5.7+0.6%. It improved to 11.2+0.9% post-exercise. There was no statistically significant weight loss noted between the 2 groups, however, biochemistry showed significant reduction in leptin, IL-6, TNF[alpha], hs-CRP, Triglyceride, and ApoB levels. Insulin sensitivity (HOMA) and ApoA1 improved post exercise. CD34/VEGFR2 + cells increased post-exercise. EPC gene expression analysis showed decrease of eNOS (5 fold) and increase of PECAM-1 (4-fold), Endothelin-1 (3-fold), IL-6 (2-fold), TNF[alpha] (3-fold) in non-exercise which improved significantly following exercise, with no change in von-Willebrand[apos]s factor. EPC migration improved post exercise, particularly in response to SDF-1[alpha].[br]Conclusion: In this pilot study we demonstrate that pre-diabetic state is associated with poor vascular reactivity and impaired endothelial progenitor cell function. Pre-diabetes may be a clinical window of therapeutic intervention opportunity when intervention such as aerobic exercise allows significant improvement of vascular reactivity, endothelial inflammation and EPC function despite no statistically significant weight loss.[br][br]Nothing to Disclose: PS, MY, AL, CC, SW, AI, SS 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3076 204 1754 SUN-LB22 PO02_LB Sunday 1551 2012


1548 ENDO12L_SUN-LB23 POSTER SESSION: Late Breaker Poster Session (1:30 PM-3:30 PM) No Increased Mortality, Cancer or Fracture Incidence in Late Postmenopausal PCOS Women [ndash] A 21-Year Controlled Follow-Up Study Johanna Schmidt, Kerstin Landin-Wilhelmsen, Mats Br[auml]nnstr[ouml]m, Eva Dahlgren Sahlgrenska University Hospital at Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden The PolyCystic Ovary Syndrome (PCOS) affects approximately 10% of women. It is characterized by oligo/anovulation, hyperandrogenism and PCO morphology. PCOS is associated with infertility, acne, hirsutism, abdominal obesity, type 2 diabetes, hypertension and dyslipidemia. Hence, women with PCOS carry several cardiovascular (CVD) risk factors.[br]The aim was to investigate women with PCOS during 21 years follow-up in the postmenopausal ages concerning anthropometry, reproductive hormones, bone mineral density, fractures, CVD risk factors and events and total mortality.[br]PCOS women, n=35, 61-79 years, diagnosed during 1956-65 and their 120 randomly allocated age-matched controls, (from the WHO MONICA study, Gothenburg), were examined in 1987 regarding anthropometry, reproductive hormones, blood pressure, bone mineral density, lifestyle factors, medications, diseases and re-examined 21 years later in 2008 regarding the same variables. The women with PCOS were still, in 2008, similar to controls regarding age and BMI, despite uncontrolled drop-out during the 21 years of follow-up.[br]Women with PCOS had persistingly higher free androgen index, but lower FSH and SHBG than controls. Hirsutism, hypertension and hypertriglyceridemia were more common, but climacteric symptoms and hypothyroidism were less prevalent among PCOS. The higher waist circumference among women with PCOS in 1987 had disappeared at follow-up, possibly due to an increase in hip circumference in PCOS and to an increase in waist circumference among controls in 2008. Bone mineral density, fractures, diabetes, CVD events, total mortality and cancer incidence were similar in the PCOS women and controls at follow-up.[br]In conclusion, late postmenopausal women with PCOS were still hyperandrogenic and hirsute with persistent hypertension and hypertriglyceridemia. However, the incidence of fractures, diabetes, cancer, CVD morbidity and total mortality was similar to the population at 21 years follow-up.[br][br]Nothing to Disclose: JS, KL-W, MB, ED 2012-06-24T13:30:00 Expo 2012-06-24T00:00:00 1899-12-30T13:30:00 3094 204 1755 SUN-LB23 PO02_LB Sunday 1552 2012


1549 ENDO12L_OR20-1 ORAL SESSION: Adipose Tissue Development [amp] Inflammation (11:15 AM-12:45 PM) Impaired Thermogenesis and Development of Brown Adipose Tissue in Mice with Combined Disruption of Insulin and IGF-1 Signaling in Fat Jeremie Boucher, Marcelo A Mori, Kevin Y Lee, Chong Wee Liew, Matthias Blueher, Steven Russell, C Ronald Kahn Joslin Diabetes Center, Boston, MA; University Hospital Leipzig, Leipzig, Germany Insulin and insulin like growth factor 1 (IGF-1) signaling play important roles in adipocyte differentiation, glucose tolerance and insulin sensitivity. Since these pathways provide similar signals and can compensate for each other in effects on adipose tissue and glucose homeostasis in vivo, we created mice with a double tissue specific knockout of both insulin receptor (IR) and IGF-1 receptor (IGF1R) by breeding mice with IR and IGF1R floxed alleles with aP2-Cre mice to eliminate all insulin/IGF-1 signaling in fat. On normal chow diet, these FIGIRKO mice were significantly leaner than control mice with a 50% decrease in white fat mass, despite no change in food intake. FIGIRKO mice were also completely resistant to high fat diet (HFD) induced obesity and were protected against age-related and HFD-induced glucose intolerance. Energy expenditure was increased in FIGIRKO mice compared to controls, despite a [gt]85% reduction in brown adipose tissue (BAT) mass. This was due to a failure of brown preadipocytes to differentiate. In vitro, IR and IGF1R knockout brown preadipocytes showed a complete impairment in their ability to differentiate into adipocytes, even in the presence of PPAR[gamma] agonist rosiglitazone. C/EBP[alpha] and PPAR[gamma] expression increased steadily in control cells during differentiation, but not in knockout cells. However, C/EBP[beta] and C/EBP[delta] expression pattern was normal but C/EBP[beta] phosphorylation was strongly decreased in double knockout cells, indicating that insulin and IGF-1 signaling are essential for C/EBP[beta] activation. Body temperature was similar between control and FIGIRKO mice at room temperature, but FIGIRKO mice were unable to maintain temperature when exposed to a 4[deg]C environment, with a drop of more than 10[deg]C with a 2 hr cold challenge. Brown fat activity on PET scanning was also markedly decreased in FIGIRKO mice, but was still responsive to [beta]3-agonist stimulation. Interestingly, a chronic [beta]3-agonist treatment led to the recruitment of systemic brown adipocytes in white adipose tissue depots of both control and FIGIRKO mice, suggesting that insulin/IGF-1 signaling is essential for discrete or preformed interscapular BAT development but not systemic brown adipocyte formation. Thus, insulin/IGF-1 signaling play a crucial role in the control of brown and white fat development, which when disrupted can lead to a defect in adaptative thermogenesis and a paradoxical increase in basal metabolic rate despite reduced brown fat mass and function.[br][br]Nothing to Disclose: JB, MAM, KYL, CL, MB, SR, CRK 2012-06-25T11:15:00 320 2012-06-25T00:00:00 1899-12-30T11:15:00 2241 267 1869 OR20-1 OR27-01 Monday 1553 2012


1550 ENDO12L_OR20-2 ORAL SESSION: Adipose Tissue Development [amp] Inflammation (11:15 AM-12:45 PM) Berardinelli-Seip Congenital Lipodystrophy 2/Seipin Is a Cell-Autonomous Regulator of Lipolysis Essential for Adipocyte Differentiation Weiqin Chen, Benny Chang, Pradip Saha, Sean M Hartig, Lan Li, Vasumathi Theegala Reddy, Yisheng Yang, Vijay Yechoor, Michael A Mancini, Lawrence Chan Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; St Luke[apos]s Episcopal Hospital, Houson, TX Mutations in BSCL2 underlie human congenital generalized lipodystrophy. We inactivated Bscl2 in mice to examine the mechanismswhereby absence of Bscl2 leads to adipose tissue loss and metabolic disorders. Bscl2/mice develop severe lipodystrophyof white adipose tissue (WAT), dyslipidemia, insulin resistance, and hepatic steatosis. In vitro differentiation of both Bscl2/murine embryonic fibroblasts (MEFs) and stromal vascular cells (SVCs) reveals normal early-phase adipocyte differentiation but a striking failure in terminal differentiation due to unbridled cyclic AMP (cAMP)-dependent protein kinase A (PKA)-activated lipolysis, which leads to loss of lipid droplets and silencing of the expression of adipose tissue-specific transcription factors. Importantly, such defects in differentiation can be largely rescued by inhibitors of lipolysis but not by a gamma peroxisome proliferator-activated receptor (PPAR) agonist. The residual epididymal WAT (EWAT) in Bscl2/mice displays enhanced lipolysis. It also assumes a [ldquo]brown-like[rdquo] phenotype with marked upregulation of UCP1 and other brown adipose tissue-specific[ markers. Together with decreased Pref1 but increased C/EBP levels, these changes highlight a possible increase in cAMP signaling that impairs terminal adipocyte differentiation in the EWAT of Bscl2/ mice. Our study underscores the fundamental role of regulated cAMP/PKA-mediated lipolysis in adipose differentiation and identifies Bscl2 as a novel cell-autonomous determinant of activated lipolysis essential for terminal adipocyte differentiation.[br][br]Sources of Research Support: Grant HL-51586 (to L.C.) and grant P30DK079638 American Heart. Association.[br][br]Nothing to Disclose: WC, BC, PS, SMH, LL, VTR, YY, VY, MAM, LC 2012-06-25T11:30:00 320 2012-06-25T00:00:00 1899-12-30T11:30:00 860 267 1870 OR20-2 OR27-01 Monday 1554 2012


1551 ENDO12L_OR20-3 ORAL SESSION: Adipose Tissue Development [amp] Inflammation (11:15 AM-12:45 PM) Brown Adipocyte Differentiation Is Regulated by Hedgehog Signaling during Development Miao-Hsueh Chen, Lagina M Nosavanh Baylor College of Medicine, Houston, TX During development, brown fat tissue arises from mesenchymal precursor cells under the control of signaling networks that are not yet well understood. The Hedgehog (Hh) signaling pathway is one of the major signaling pathways that regulate mesenchymal cell fate. However, whether the Hh pathway controls the formation of brown fat is not clear. To investigate the role of the Hh pathway in brown fat development, we selectively activated the Hh pathway in fat tissue by mating the constitutively active M2 allele of Smo (SmoM2) to aP2-Cre transgenic mice, which express the Cre recombinase in many embryonic tissues including brown fat tissue. Phenotypic analyses indicate that aP2-Cre; SmoM2 mice die at birth with multiple tissue defects including polydactyly and open neural tubes, the two classic phenotypes associated with Hh pathway gain-of-function. Most importantly, our analyses showed that the amount of brown fat in these mutants is significantly reduced, suggesting that the Hh pathway can negatively regulate brown fat development. To confirm our in vivo observations and to establish that Hh signaling can directly control brown preadipocyte differentiation, we isolated and immortalized brown preadipocytes from E16.5 wild-type embryos. We first examined whether the Hh pathway components are present in these brown preadipocytes. Our results indicate that Smo and the downstream transcription factors Gli2 and Gli3 are present in these cells and localize to the primary cilium, the Hh signaling center, upon exogenous Hh stimulation. Furthermore, we found that the primary cilium, which is responsible for the activation and processing of the Gli transcription factors, is only present in brown preadipocytes but not in fully differentiated brown adipocytes. Finally, our studies indicate that applying Shh ligand together with adipogenic induction media can inhibit the differentiation of these brown preadipocytes. Specifically, Hh treatment resulted in significantly less accumulation of lipid droplets and in the down-regulation of the expression of pro-adipogenic genes. Taken together, our studies support an inhibitory role for Hh signaling in brown fat development.[br][br]Nothing to Disclose: M-HC, LMN 2012-06-25T11:45:00 320 2012-06-25T00:00:00 1899-12-30T11:45:00 571 267 1871 OR20-3 OR27-01 Monday 1555 2012


1552 ENDO12L_OR20-4 ORAL SESSION: Adipose Tissue Development [amp] Inflammation (11:15 AM-12:45 PM) Genetic Disruption of a Novel Open Reading Frame in Mice Results in Skin and Adipose Tissue Abnormalities and Uncovers a Link between Adipocytes and Keratinocytes Jun Zhu, Farid Chehab University of California, San Francisco, CA The delineation of causative genes affecting obese patients with rare congenital disorders can provide novel clues into mechanisms which link adiposity to other systems that thrive on metabolism. In this context, we characterized the 7:22 chromosome translocation breakpoint of an obese patient with Coffin-Siris syndrome, a rare genetic disorder associated with feeding difficulties, mental retardation and coarse features. Uncovering of the translocation breakpoint on chromosome 7q by molecular, cytogenetic and array CGH studies revealed interruption of a novel open reading frame gene highly conserved in vertebrates and with no known function. mRNA expression by real-time PCR and Western blot analysis of the encoded protein revealed widespread expression in mouse tissues and secretion into the plasma and cerebrospinal fluid in humans. To gain further insights into the function of this gene, we generated mice with an inducible Cre-Lox cassette targeted to one of its exons. Crossing of these floxed mice with transgenic mice expressing Cre recombinase under control of the ubiquitous cytomegalovirus promoter allowed us to generate total body knockout mice. Homozygous mice exhibited embryonic lethality before 12.5 days of gestation whereas heterozygous mice were viable. However, 20% of heterozygous knockout mice die within two weeks of birth and display a phenotype that involves the adipose fat mass and skin. The predominant features of these knockout mice are evident a week after birth and include hypopigmentation, alopecia, a thick skin with hyperkeratosis and a normal skin barrier function. The basal layer of epidermal keratinocytes exhibited a significant increase in proliferation as determined by immunostaining of skin sections from knockout mice with anti-Ki67, a well-known proliferation marker. As a result, the adipocyte layer in the hypodermis was greatly reduced. Furthermore, these mice show a 30% reduction in brown adipose fat mass and complete absence of subcutaneous interscapular white adipose tissue compared to large depots in littermate controls. Overall, we hypothesize that mobilization of energy stores from adipocytes is shunted to the skin to sustain the energy required for the proliferation of keratinocytes. Thus, unveiling of the metabolic link between adipocytes and keratinocytes will allow characterization of the novel axis between subcutaneous adipose tissue and skin.[br][br]Sources of Research Support: A grant from the UCSF Diabetes and Endocrinology Research Center.[br][br]Nothing to Disclose: JZ, FC 2012-06-25T12:00:00 320 2012-06-25T00:00:00 1899-12-30T12:00:00 1878 267 1872 OR20-4 OR27-01 Monday 1556 2012


1553 ENDO12L_OR20-5 ORAL SESSION: Adipose Tissue Development [amp] Inflammation (11:15 AM-12:45 PM) Ablation of Ghrelin Receptor Attenuates High-Fructose Corn Syrup (HFCS)-Induced Adipose Inflammation and Insulin Resistance Jing Yue, Xiaojun Ma, Ligen Lin, Huaizhu Wu, Clifton W Smith, Yuxiang Sun Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX Adipose inflammation and insulin resistance play causal roles for type 2 diabetes. High fructose corn syrup (HFCS) is the most-used sweetener in the United States, substantially replacing sucrose. Some studies have suggested that HFCS consumption correlates with obesity and insulin resistance, while other studies are in disagreement. Due to conflicting and insufficient scientific evidence, HFCS continues to be used as the primary sweetener in our food supplies.[br]In this study, we compared the metabolic effects of mice fed regular diet, high fat diet, or regular diet supplemented with 8% HFCS in drinking water (to mimic soft drinks). We found that high fat diet-fed mice consumed the most total calories, and showed the most weight gain and fat deposition. Surprisingly and counter-intuitively, HFCS-fed mice exhibited the most severe insulin resistance; disproportionately greater in relation to their calorie intake and total body fat content. Adipose tissue macrophages (ATMs) have emerged as a major pathogenic factor for obesity and insulin resistance. ATMs consist of pro-inflammatory F4/80[sup]+[/sup]/CD11c[sup]+[/sup] macrophages and anti-inflammatory F4/80[sup]+[/sup]/CD11c[sup]-[/sup] macrophages. Like high fat diet, HFCS induced robust increases of both pro-inflammatory and anti-inflammatory ATMs in visceral fat, but the number of anti-inflammatory ATMs was much lower in HFCS-fed mice than in high fat-fed mice.[br]Ghrelin is an orexigenic hormone, promoting adiposity and insulin resistance. The biologically relevant receptor of ghrelin is Growth Hormone Secretagogue Receptor (GHS-R). To elucidate whether ghrelin/GHS-R signalling affects macrophages in adipose tissues, we investigated the effects of GHS-R in HFCS-induced adipose inflammation, liver steatosis and insulin resistance using our [italic]Ghsr[/italic]-null mice. The [italic]Ghsr[/italic]-null mice exhibited lower pro-inflammatory ATMs and pro-inflammatory cytokine expression, but no difference in anti-inflammatory ATMs, in visceral fat. Moreover, the [italic]Ghsr[/italic]-null mice showed attenuated liver steatosis and less-pronounced HFCS-induced insulin resistance. [italic]In vitro[/italic], our data further revealed that GHS-R gene knockdown in macrophage RAW264.7 cells resulted in reduced pro-inflammatory cytokines, suggesting that GHS-R has direct effect on macrophages. Thus, HFCS consumption has additional detrimental effects on insulin resistance beyond the extra calories from HFCS, and GHS-R antagonists may represent novel drugs for ameliorating adipose inflammation and insulin resistance.[br][br]Sources of Research Support: We gratefully acknowledge the support of NIH/NIA grant 1R03AG029641-01 (YS), USDA/CRIS grant ARS 6250-51000-055 (YS), and the NIH-Diabetes and Endocrinology Research Center (P30DK079638) at Baylor College of Medicine.[br][br]Nothing to Disclose: JY, XM, LL, HW, CWS, YS 2012-06-25T12:15:00 320 2012-06-25T00:00:00 1899-12-30T12:15:00 1377 267 1873 OR20-5 OR27-01 Monday 1557 2012


1554 ENDO12L_OR20-6 ORAL SESSION: Adipose Tissue Development [amp] Inflammation (11:15 AM-12:45 PM) A Crosstalk between Adipocytes and Adipose T-Cells in the Initial Phase of Obesity Induced Inflammation Tuo Deng, Jianxin Lin, Laurie Minze, Patrick Reardon, Vadim Sherman, Rongfu Wang, Christopher Lyon, Willa Hsueh The Methodist Hospital Research Institute, Houston, TX; The Methodist Hospital, Houston, TX; The Methodist Hospital Research Institute, Houston, TX Increasing evidence suggests that T-cell activation in adipose tissue during high fat diet (HFD) is involved in obesity-induced inflammation. It is unknown what initiates adipose tissue T-cell activation and infiltration, but adipocytes are a logical source of regulatory signals. In adipocytes isolated from subcutaneous and visceral adipose tissue obtained from elective surgeries of obese and lean post-menopausal women, we found that class II transactivator (CIITA), the master regulator of major histocompatibility complex class II (MHCII) antigen presentation, and multiple MHCII family genes, were substantially increased with obesity. Flow cytometry also detected more MHCII+ adipocytes in obese versus lean subjects (10.6% vs. 2.6%, p[lt]0.01). Similar patterns were found in obese mouse models: db/db, high-fat diet (HFD)-fed LDLR[sup]-/-[/sup] mice, and C57Bl/6 mice given 60% lard diet, in which adipocyte leptin increases after one week HFD and adipocyte MHCII and SVF leptin increase at two weeks. Among inflammatory factors increased in adipose during obesity, only IFNg dramatically induced adipocyte MHCII gene expression. Leptin increases T-cell IFNg secretion suggesting that IFN[gamma] from the adipose stromal vascular fraction (SVF), which contains T-cells, contributes to this early increase in MHCII. SVF IFNg and adipocyte MHCII expression of leptin-deficient ob/ob mice are much less than HFD-fed obese mice, supporting the idea that leptin is a major regulator of adipose T-cell IFNg and subsequent adipocyte MHCII expression. Finally, we found that both primary and 3T3-L1 adipocytes could stimulate T-cell activation and Th1 polarization in an antigen-specific, contact-dependent manner, which could be attenuated by CIITA-knockdown in 3T3-L1 adipocytes or by use of MHCII-deficient primary adipocytes. In addition, adipocytes from obese mice induced dramatically more T-cell activation and Th1 polarization than those of lean mice. In summary, our study suggests an interesting a crosstalk between adipocytes and adipose T-cells in the initial phase of HFD-induced inflammation. Leptin is induced in expanded adipocytes, increases IFNg production in adipose resident T-cells, which induces MHCII genes in adipocytes. This crosstalk further activates adipose T-cell responses to amplify inflammation in obese adipose tissue.[br][br]Nothing to Disclose: TD, JL, LM, PR, VS, RW, CL, WH 2012-06-25T12:30:00 320 2012-06-25T00:00:00 1899-12-30T12:30:00 2360 267 1874 OR20-6 OR27-01 Monday 1558 2012


1555 ENDO12L_OR21-1 ORAL SESSION: Novel Factors Controlling Islet Cell Metabolism (11:15 AM-12:45 PM) Basal Hyperglucagonemia Is Associated with Defects in Glucagon Counterregulation in Type 1 Diabetes Leon S Farhy, Alice Chan, Marc D Breton, Stacey M Anderson, Boris P Kovatchev, Anthony L McCall University of Virginia, Charlottesville, VA; University of Virginia, Charlottesville, VA Glucagon Counterregulation (GCR) is a key protection against hypoglycemia, which is impaired in type 1 diabetes (T1DM) by an unknown mechanism. Our model-based analysis of in vivo animal data predicts that the GCR defects are linked to basal hyperglucagonemia and the goal of this work is to test this hypothesis on clinical data. To this end, we analyzed the relationship between basal glucagon (BasG) during euglycemia and the GCR response to hypoglycemia in 29 hyperinsulinemic hypoglycemic clamps in T1DM patients. Glucose levels were stabilized in euglycemia and then steadily lowered to 50 mg/dL. Glucagon was measured before induction of hypoglycemia and at 10 min intervals after glucose reached levels below 70 mg/dL. Individual GCR was assessed by RIG, the relative increase in glucagon over basal. Analysis of the results was performed with our mathematical model of GCR (e.g., see (1)). This model describes interactions between islet peptides and circulating glucose, reproduces the normal GCR axis and its impairment in diabetes. It was used to identify a control mechanism consistent with the observed data linking between BasG and GCR. BasG levels were 40.5 [plusmn] 21.9 pg/mL and RIG was 118 [plusmn] 112.7 % (mean [plusmn] standard deviation). We detected that higher BasG was associated with lower GCR response and, in particular, RIG correlated negatively with BasG (r = -0.74, p [lt] 0.0001). Consistent with these results was a model of GCR in which the secretion of glucagon has two components. The first is under (auto) feedback control and drives a pulsatile GCR and the second is feedback independent (basal secretion) and its increase suppresses the GCR. Model-based simulations showed that such auto-feedback control of GCR can explain the observed relationships between BasG and RIG during a 3-fold simulated increase in BasG. Further increase obliterated the ability of the system to counterregulate. Our findings support the hypothesis that basal hyperglucagonemia contributes to the GCR impairment in T1DM and show that the predictive power of our GCR model applies to human pathophysiology in T1DM. They also emphasize the need to test clinically the model-based prediction that glucagon inhibitors may enhance GCR in T1DM (1). If confirmed, this could lead to novel clinically relevant strategies for treatment of T1DM with enhanced protection against hypoglycemia.[br][br](1) Farhy LS, McCall AL. Optimizing Reduction in Basal Hyperglucagonaemia to Repair Defective Glucagon Counterregulation in Insulin Deficiency. Diabetes, Obesity and Metabolism 13 (s1), 133[ndash]143, 2011.[br][br]Sources of Research Support: RO1 DK082805, RO1 DK51562, UVa GCRC M01 RR 000847.[br][br]Nothing to Disclose: LSF, AC, MDB, SMA, BPK, ALM 2012-06-25T11:15:00 372 2012-06-25T00:00:00 1899-12-30T11:15:00 1242 268 1875 OR21-1 OR13-01 Monday 1559 2012


1556 ENDO12L_OR21-2 ORAL SESSION: Novel Factors Controlling Islet Cell Metabolism (11:15 AM-12:45 PM) Osteocalcin Regulates Perinatal [beta]-Cell Proliferation through the Gprc6a Receptor Jianwen Wei, Nina Suda, Gerard Karsenty, Patricia Ducy Columbia University, New York, NY; Columbia University, New York, NY During the perinatal period [beta]-cells undergo a transient peak of proliferation that significantly contributes to the rapid increase in [beta]-cell mass observed postnatally. Remarkably, the onset of this perinatal peak of [beta]-cell proliferation coincides with de novo expression by the developing embryo of osteocalcin, a bone-derived hormone known to increase insulin secretion and [beta]-cell proliferation in adult pancreas. Analysis of osteocalcin-deficient pups revealed that [beta]-cell proliferation is significantly decreased between P0 and P10 in absence of osteocalcin indicating that this hormone is a molecular determinant of perinatal [beta]-cell proliferation. To determine if this regulatory mechanism is mediated by Gprc6a, a G-protein coupled receptor mediating another function of osteocalcin, we used the Pdx1-Cre deleter strain to inactivate Gprc6a specifically in the [beta]-cell lineage. This conditional inactivation resulted in a low rate of perinatal [beta]-cell proliferation similar to the one observed in pups deficient in osteocalcin, and led to glucose intolerance and impaired insulin secretion in response to glucose in young mice. A more detailed analysis of the phenotype of the Gprc6aPdx1-/- mutant mice will be presented at the meeting. These studies identify osteocalcin, signaling through the Gprc6a receptor expressed in [beta]-cells, as an endocrine determinant of the perinatal peak of [beta]-cell proliferation.[br][br]Sources of Research Support: JDRF grant to PD.[br][br]Nothing to Disclose: JW, NS, GK, PD 2012-06-25T11:30:00 372 2012-06-25T00:00:00 1899-12-30T11:30:00 1140 268 1876 OR21-2 OR13-01 Monday 1560 2012


1557 ENDO12L_OR21-3 ORAL SESSION: Novel Factors Controlling Islet Cell Metabolism (11:15 AM-12:45 PM) GLP-2 Receptor in Endocrine Cells Is Required for Glucose Homeostasis Xuemei Shi, Benny Chang, Lawrence Chan, Xinfu Guan Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX Glucagon-like peptide-2 (GLP-2), GLP-1 and peptide YY are co-secreted from endocrine L cells in response to food intake; and play a key role in the control of glucose homeostasis. GLP-2 exerts diverse actions in the gastrointestinal tract, including enhancing epithelial cell survival and proliferation, mucosal blood flow, and nutrient uptake and suppressing gastric motility and secretion. These actions are mediated by the G-protein-coupled receptor (GLP-2R). However, little is known about its physiological role in the control of glucose homeostasis. GLP-2R expressed in enteroendocrine cells may modulate their function and glucose metabolism. We hypothesize that GLP-2R deletion specifically in enteroendocrine cells impairs glucose homeostasis and insulin sensitivity. To address this hypothesis, we generated mice lacking [italic]Glp2r[/italic] in enteroendocrine cells with the Cre-LoxP approach, i.e., [italic]Glp2r[/italic][sup]lox/lox[/sup] mice were crossed with villin cre mice. Villin cre[sup]+/0[/sup], [italic]Glp2r[/italic][sup]lox/lox[/sup] knockout (KO) mice displayed impaired glucose homeostasis though body weight and basal glucose were normal. In response to oral glucose tolerance test, villin-[italic]Glp2r[/italic] KO displayed decelerated glucose clearance compared to their wild-type littermates. We further quantified glucose homeostasis using [sup]2[/sup]H[sub]2[/sub]O and D-6,6-[sup]2[/sup]H[sub]2[/sub]-glucose stable isotopic tracers coupled with hyperinsulinemic euglycemic clamp. Villin-[italic]Glp2r[/italic] KO mice showed decreases in glucose infusion rate (KO vs WT: 3.60 [plusmn] 0.74 vs 6.93 [plusmn] 0.97 mmol/kg/h, [italic]p [/italic][lt] 0.01) and glucose disappearance rate (KO vs WT: 8.69 [plusmn] 1.05 vs 12.01 [plusmn] 1.38 mmol/kg/h, [italic]p [/italic][lt] 0.07). However, villin-[italic]Glp2r[/italic] KO mice displayed normal hepatic glucose production (KO vs WT: 5.09 [plusmn] 0.97 vs 5.09 [plusmn] 1.28 mmol/kg/h) and gluconeogenesis (KO vs WT: 2.74 [plusmn] 0.59 vs 2.58 [plusmn] 0.79 mmol/kg/h). Thus, we conclude that [italic]Glp2r[/italic] deletion in endocrine cells in the gut results in whole-body insulin resistance mainly due to decreased glucose uptake and utilization by peripheral tissues. Therefore, GLP-2R signaling in enteroendocrine cells is required for the maintenance of glucose homeostasis at high postprandial insulin.[br][br]Sources of Research Support: NIH Grant DK075489 and DK084125 awarded to XG; USDA Grant 6250-51000-043 awarded to XG.[br][br]Nothing to Disclose: XS, BC, LC, XG 2012-06-25T11:45:00 372 2012-06-25T00:00:00 1899-12-30T11:45:00 975 268 1877 OR21-3 OR13-01 Monday 1561 2012


1558 ENDO12L_OR21-4 ORAL SESSION: Novel Factors Controlling Islet Cell Metabolism (11:15 AM-12:45 PM) Cholecystokinin Expression in Pancreatic Islets Is a Protective Factor for [beta]-Cell Survival Sirinart Sirinvaravong, Jeremy A Lavine, Louise M Meske, Kathyrn L Schueler, Dawn B Davis University of Wisconsin, Madison, WI; University of Wisconsin, Madison, WI Cholecystokinin (CCK) is expressed in pancreatic islet and is highly upregulated in obesity. Absence of CCK leads to decreased [beta]-cell mass due to increased apoptosis. In vitro, addition of CCK was able to rescue apoptosis from ER-stress in CCK knockout islets[1].[br]To determine if over expression of CCK is sufficient to prevent [beta]-cell death and improve glycemic control, we generated a transgenic mouse with CCK gene expression driven by the mouse insulin promotor (MIP). No differences were seen in random glucose and insulin levels in young, lean animals. Therefore, we proceeded to examine animals where we expected increased [beta]-cell apoptosis.[br]As animals age, basal rates of [beta]-cell apoptosis increase and there is a decline in [beta]-cell mass. We saw no consistent difference in random glucose, insulin or weight when comparing lean MIP-CCK and wild type mice up to 48 weeks of age. However, we also wanted to examine [beta]-cell mass in these older animals. Pancreata from seven MIP-CCK and seven wild type mice were studied (age 12 to 15 months). We quantitated fractional [beta]-cell area as a percentage of total pancreatic sectional area using immunofluorescense staining with an anti-insulin antibody. MIP-CCK mice had higher fractional [beta]-cell area (5.07[plusmn]1.64% vs 1.38 [plusmn] 0.41 %, p [lt]0.05) compared with wild type mice.[br]To examine the effects of MIP-CCK overexpression in a model of more pronounced [beta]-cell apoptosis, we treated 10 to 13 week-old mice with multiple low doses of the [beta]-cell toxin streptozotocin (STZ).We found that MIP-CCK mice are resistant to STZ-induced diabetes, as measured by fasting glucose values (186 [plusmn] 51 mg/dL vs. 264 [plusmn] 71 mg/dL at 21 days after STZ, p[lt]0.05).[br]In summary, our data show that overexpression of CCK is protective in models of insulin resistance and [beta]-cell death. We believe this is due to the ability of CCK to protect [beta]-cells from apoptosis, thus improving [beta]-cell mass and leading to improved glycemic control.[br][br]Lavine, J.A., et al., Cholecystokinin Is Up-Regulated in Obese Mouse Islets and Expands {beta}-Cell Mass by Increasing {beta}-Cell Survival. Endocrinology, 2010. 151(8): p. 3577-88.[br][br]Sources of Research Support: NIDDK K08DK3442 awarded to DBD; University of Wisconsin Graduate School and School of Medicine and Public Health.[br][br]Nothing to Disclose: SS, JAL, LMM, KLS, DBD 2012-06-25T12:00:00 372 2012-06-25T00:00:00 1899-12-30T12:00:00 1698 268 1878 OR21-4 OR13-01 Monday 1562 2012


1559 ENDO12L_OR21-5 ORAL SESSION: Novel Factors Controlling Islet Cell Metabolism (11:15 AM-12:45 PM) High-Fat Diet during Perinatal Development Reduces [alpha]-Cell Mass and Leads to Early Insulin Resistance in the Infantile Nonhuman Primate Sarah M Comstock, Jacalyn M Bishop, Diana L Takahashi, Lynley Pound, Kevin L Grove OHSU/ONPRC, Beaverton, OR The primary purpose of this study was to investigate the impact of maternal high fat diet (HFD) exposure during pregnancy and to determine if this will affect fetal islet morphology and lead to physiologic changes in the infant monkey. Fetuses were collected in the early 3rd trimester from Japanese macaques that were placed on a control (CTR) or HFD diet for 4 years. Pancreata were processed for gene expression and immunohistochemistry. Physiological measurements, including body weight, body fat percentage, fasting glucose, insulin, glucagon and response to an i.v. GTT were collected from infantile offspring from CTR and HFD mothers up to 210 days of age. While circulating insulin was unchanged, c-peptide levels were decreased, indicating that HFD offspring have decreased insulin clearance. Analysis of islet cells demonstrated that although total islet mass and [beta]-cell mass were not changed in the fetal HFD offspring, [alpha]-cell mass was decreased. This led to an increase in the [beta]-cell/[alpha]-cell ratio in HFD offspring, which correlated with placental glucose levels and hepatic gluconeogenic gene expression. These animal display a significant change in expression of genes involved in glucose homeostasis and islet neogenesis, including PDX1, NeuroD, Glucokinase and Glut2. The initial stages in the glucose stimulated insulin secretion pathway were up-regulated, while insulin secretion was decreased (as measured by c-peptide levels). HFD infant offspring had decreased insulin secretion at 90 days of age, however, by 180days insulin secretion was normalized. These HFD offspring were also significantly heavier, had increased adiposity and were significantly insulin resistant by 7 months of age. These studies demonstrate that HFD consumption during pregnancy can lead to an adaptive [alpha]-cell response that affects the paracrine regulation of fetal islets. This allows animals to maintain normoglycemia after birth. However, the post-natal consumption of the HFD ultimately produces decreased insulin sensitivity.[br][br]Sources of Research Support: NIH grants DK-060685 (to K.L. Grove), DK-079194 (to K.L. Grove), and RR000163 (to K.L. Grove and ONPRC).[br][br]Nothing to Disclose: SMC, JMB, DLT, LP, KLG 2012-06-25T12:15:00 372 2012-06-25T00:00:00 1899-12-30T12:15:00 2373 268 1879 OR21-5 OR13-01 Monday 1563 2012


1560 ENDO12L_OR21-6 ORAL SESSION: Novel Factors Controlling Islet Cell Metabolism (11:15 AM-12:45 PM) A Glucagon-Like Peptide-1-Estrogen Fusion Peptide Shows Enhanced Efficacy in Preventing Insulin-Deficient Diabetes in Mice Joseph P Tiano, Brian Finan, Richard DiMarchi, Franck Mauvais-Jarvis Northwestern University, Chicago, IL; Indiana University, Bloomington, IN Preserving functional [beta]-cell mass in diabetes mellitus is a major therapeutic challenge. The female hormone 17[beta]-estradiol (E2) protects islet [beta]-cells from apoptosis [italic]in vivo[/italic] in mice and in cultured human islets via activation of estrogen receptors (ERs). This raises the prospect that ER activation is a therapeutic avenue to protect [beta]-cell mass. However, the risk of hormone dependent cancer precludes the use of estrogens as a therapy for diabetes. To preferentially target E2 to the [beta]-cells but without the undesirable effect of general E2 therapy, we created novel fusion peptides combining glucagon-like peptide-1 (GLP-1) and E2 in a single molecule. By marrying the pharmacologies of GLP-1 and E2, we envisioned a synergistic action on glycemic control by the combined insulinotropic and anti-apoptotic activities on pancreatic [beta]-cells that express ERs and GLP-1 receptors (GLP-1R). Two conjugates were synthesized with E2 stably linked to GLP-1 to avoid E2 release in the circulation and maximize E2 release in target cells by virtue of proteolysis: an active GLP-1 stably linked to E2 (aGLP1-E2) and an inactive GLP-1 stably linked to E2 (iGLP1-E2). This later conjugate binds GLP-1R normally but is pharmacologically inactive and incapable of activating GLP-1R signaling and is a unique pharmacological tool to direct E2 to [beta]-cells. All conjugates efficiently protected cultured mouse islets from oxidative stress-induced apoptosis demonstrating that GLP-1 efficiently directed E2 to intracellular ERs. We tested the efficiency of GLP1-E2 conjugates in preventing [beta]-cell destruction in the model of multiple low-dose injection of streptozotocin (STZ). As we previously described, general E2 therapy prevented STZ-induced insulin-deficient diabetes. We observed that the aGLP-1 used at low dose (120mg/kg) reduced the severity of STZ-induced diabetes. Importantly, the iGLP1-E2 conjugate prevented STZ-induced insulin deficient diabetes demonstrating that the inactive GLP-1 was able to bind the GLP-1R and to direct E2 to the [beta]-cells for protection [italic]in vivo. [/italic]Most importantly the aGLP1-E2 conjugate was more potent than the aGLP-1 and the iGLP1-E2 individually in preventing insulin deficient diabetes. All conjugates were devoid of E2 gynecological effect compared to general E2 therapy. These observations provide the proof of concept, that combining GLP-1 and E2 in a single molecule provides synergism in protecting [beta]-cell survival without the side effect of general estrogen therapy.[br][br]Sources of Research Support: Research supported by NIH grants RO1 DK074970-01, P50 HD044405, T32 DK007169.[br][br]Nothing to Disclose: JPT, BF, RD, FM-J 2012-06-25T12:30:00 372 2012-06-25T00:00:00 1899-12-30T12:30:00 1969 268 1880 OR21-6 OR13-01 Monday 1564 2012


1561 ENDO12L_OR22-1 ORAL SESSION: Novel Mechanisms in Lipid Metabolism (11:15 AM-12:45 PM) Brain Insulin Signaling Increases Hepatic Triglyceride Secretion [italic]In Vivo[/italic] Thomas Scherer, Claudia Lindtner, James O[apos]Hare, Liz Zielinski, Ludger Scheja, Christoph Buettner Mount Sinai School of Medicine, New York, NY; Medical University of Vienna, Vienna, Austria; University Medical Center, Hamburg, Germany Hepatosteatosis and dyslipidemia are hallmarks of the metabolic syndrome and plasma triglycerides (TG) tightly correlate with insulin resistance (IR). Since hepatic lipogenesis is increased in the IR state, TG secretion must not be too low in order to prevent steatosis. Insulin action comprises direct effects on peripheral organs e.g. liver and adipose, but also indirect effects mediated via the central nervous system(1). Systemic insulin decreases very low-density lipoprotein (VLDL) production by the liver, yet it is unknown whether brain insulin can independently regulate VLDL flux. To study the role of brain vs. systemic insulin signaling on hepatic VLDL secretion, we performed tyloxapol infusion studies in male Sprague Dawley rats during systemic or isolated brain hyperinsulinemia. The latter was accomplished by infusing insulin or vehicle for 4 hrs into the 3rd ventricle (ICV) or the mediobasal hypothalamus (MBH). ICV insulin infusion increased hepatic VLDL secretion compared to controls (2.59[plusmn]0.28 vs. 1.80[plusmn]0.2 [mu]mol/kg/min; P=0.039; n[ge]11 per group). To the contrary, a hyperinsulinemic euglycemic clamp decreased TG flux (0.85[plusmn]0.05 [mu]mol/kg/min; P=0.020; n=4), which is in agreement with prior reports(2). Plasma lipid profiling in these rats demonstrated that ICV insulin increased the accumulation of TG associated fatty acids such as palmitate or oleate (+30%; P[lt]0.05). Of note, insulin infusion into the MBH had no effect on VLDL flux vs. controls (1.85[plusmn]0.32 [mu]mol/kg/min vs. 1.71[plusmn]0.32 [mu]mol/kg/min; P=0.773; n=5 per group) indicating that another brain region integrates the central insulin-signal. Conversely, mice that lack the insulin receptor in the whole brain had reduced hepatic TG flux compared to littermate controls, which was again assessed by tyloxapol studies (154[plusmn]6 vs. 126[plusmn]12 [mu]mol/kg/h; P = 0.038; n[ge]9 per group). To begin to understand the molecular underpinnings that alter hepatic VLDL flux when ICV insulin is infused, we assessed hepatic microsomal TG transfer protein (MTTP) expression, the rate-limiting enzyme in VLDL assembly. Consistent with VLDL flux, ICV insulin increased MTTP expression compared to controls (P=0.046; n[ge]5). While systemic hyperinsulinemia and isolated loss of neuronal insulin signaling both suppress TG flux, ICV insulin infusion acutely increases VLDL secretion. We speculate that the elevated TG production in obesity and diabetes may be due to preserved central insulin effects in a presently unknown brain region.[br][br](1) Scherer et al., Cell Metab. 2011 Feb 2;13(2):183-94. (2) Grefhorst et al., Am J Physiol Gastrointest Liver Physiol. 2005 Sep;289(3):G592-8.[br][br]Sources of Research Support: NIH Grants DK074873, DK083568 and DK082724 awarded to CB; ADA basic research award to CB; European Foundation for the Study of Diabetes Grant awarded to TS; CB is the recipient of a Hirschl Award.[br][br]Nothing to Disclose: TS, CL, JO, LZ, LS, CB 2012-06-25T11:15:00 342ABDE 2012-06-25T00:00:00 1899-12-30T11:15:00 692 269 1881 OR22-1 OR08-01 Monday 1565 2012


1562 ENDO12L_OR22-2 ORAL SESSION: Novel Mechanisms in Lipid Metabolism (11:15 AM-12:45 PM) The Autonomic Nervous System Regulates Postprandial VLDL-Triglyceride Secretion Eveline Bruinstroop, Susanne la Fleur, Ewout Foppen, Mariette Ackermans, Joke Wortel, Eric Fliers, Andries Kalsbeek Academic Medical Center (AMC), University of Amsterdam, Amsterdam, Netherlands; Academic Medical Center (AMC), University of Amsterdam, Amsterdam, Netherlands; Netherlands Institute for Neuroscience, an Institute of the Royal Netherlands Academy of Arts and Science, Amsterdam, Netherlands We have previously shown that during fasting an intact sympathetic innervation of the liver is necessary to maintain the secretion of triglycerides in very-low density lipoproteins (VLDL-TG). Interestingly, the inhibitory effect of central glucose administration on VLDL-TG secretion depends on an intact parasympathetic hepatic innervation. Together these data suggest that the activities of the antagonizing branches of the autonomic nervous system depend on nutritional state. We hypothesized that during feeding the parasympathetic input to the liver inhibits hepatic VLDL-TG secretion. We therefore determined the effect of a selective hepatic denervation on triglyceride metabolism after a meal.[br]Male Wistar rats received a surgical sham, sympathetic, parasympathetic or total liver denervation and a jugular catheter for blood sampling. After recovery to pre-surgery body weight, rats were food deprived overnight to eradicate chylomicrons from the gut and then received a chow meal challenge. Triglycerides, insulin and glucose were measured for 80 minutes after the meal. One week later we measured VLDL-TG secretion after a chow meal challenge in the same experimental animals using an intravenous bolus of tyloxapol to inhibit uptake of triglycerides by peripheral tissues.[br]All rats studied ingested a meal of 2.0-4.5 grams within 20 minutes. The meal significantly increased plasma glucose and insulin levels and decreased plasma triglyceride levels in all groups. The post-prandial triglyceride curve was significantly elevated in parasympathetic (P=0.02) and sympathetic (P=0.03) denervated rats compared to sham-operated animals. This effect was lost when both branches of the autonomic nervous system were cut (P=0.39). The post-prandial insulin and glucose curves were not significantly different between the groups. The data from the subsequent tyloxapol experiment showed that the increased post-prandial plasma triglycerides in the parasympathetic and sympathetic denervated groups can be explained by an increased VLDL-TG secretion by the liver. The effectiveness of the sympathetic denervation was evidenced by a significant decrease in liver noradrenaline levels.[br]Our experiments show that cutting either branch of the autonomic nervous system results in higher post-prandial plasma triglyceride levels as well as higher VLDL-TG secretion. This indicates that a disbalance of the autonomic input to the liver causes a disinhibition of VLDL-TG secretion after a meal.[br][br]Nothing to Disclose: EB, SlF, EF, MA, JW, EF, AK 2012-06-25T11:30:00 342ABDE 2012-06-25T00:00:00 1899-12-30T11:30:00 717 269 1882 OR22-2 OR08-01 Monday 1566 2012


1563 ENDO12L_OR22-3 ORAL SESSION: Novel Mechanisms in Lipid Metabolism (11:15 AM-12:45 PM) Transgenic Expression of HIV-1 Accessory Protein Viral Protein R (Vpr) Induces Hepatic Steatosis Neeti Agarwal, Toni Oplt, Dinakar Iyer, Ashok Balasubramanyam Baylor College of Medicine, Houston, TX Hepatic steatosis is highly prevalent in patients with HIV infection, associated with hypertriglyceridemia and lipodystrophy. The role of the HIV virus [italic]per se[/italic] in causing or exacerbating fatty liver disease is unclear. The HIV-1 accessory protein viral protein R (Vpr) acts [italic]in vitro[/italic] as a corepressor of PPAR[apos]s, critical transcription factors for fat metabolism in liver and and adipose tissues, and dysregulation of PPAR-mediated gene expression has been linked to fatty liver disease.[br]We investigated the hypothesis that Vpr can disrupt PPAR-gamma and PPAR-alpha mediated gene expression in adipose tissues and liver and lead to a fatty liver phenotype [italic]in vivo[/italic], using transgenic mice expressing Vpr under control of the PEPCK promoter. 14-week old Vpr-Tg mice or WT littermates were used for this study (8 M, 8 F in each group). Animals were fed chow with a high protein content (to stimulate the PEPCK promoter and enhance Vpr expression), and sacrificed after a 12 h fast. In the liver, expression of genes involved in fatty acid transport and fat oxidation was quantified by qPCR, and total and phosphorylated AMPK were measured by immunoblotting. Liver sections were stained with oil red O and quantified for lipid levels using ImageJ software. Lipid species in the liver were quantified by thin layer chromatography. Plasma adiponectin levels were determined by ELISA. Group results were compared using two-way paired t-tests.[br]PPAR-alpha target genes encoding fatty acid oxidative enzymes and transport proteins were downregulated in liver of Vpr-Tg mice compared to WT mice (CPT1: -23%, P=0.03; AOX: -23%, P=0.01; LCAD: -27%, P=0.04; MTP; -34%, P=0.04) by qPCR. Circulating levels of plasma adiponectin were decreased in Vpr-Tg mice (total adiponectin -30%, P=0.01; high molecular weight adiponectin -34%, P=0.02 in males and -11% in females, P=0.006). The ratio of phospho-AMPK to total AMPK was decreased in Tg-Vpr mice (-48%, P=0.01). Oil red O staining was increased in Vpr-Tg mouse livers compared to WT livers, (+250%, P=0.05). Thin layer chromatography demonstrated increased hepatic triglyceride content in Vpr-Tg mice compared to WT (+58%, P=0.0005).[br]HIV-1 Vpr disrupts fatty acid oxidation and triglyceride metabolism in liver in a manner consistent with interference with PPAR-alpha mediated gene expression, and produces a phenotype of hepatic steatosis. These data indicate a novel and significant role for Vpr in the pathogenesis of HIV asscociated fatty liver disease.[br][br]Nothing to Disclose: NA, TO, DI, AB 2012-06-25T11:45:00 342ABDE 2012-06-25T00:00:00 1899-12-30T11:45:00 2221 269 1883 OR22-3 OR08-01 Monday 1567 2012


1564 ENDO12L_OR22-4 ORAL SESSION: Novel Mechanisms in Lipid Metabolism (11:15 AM-12:45 PM) The Pro-Atherogenic Effect of Chronic Nitric Oxide (NO) Synthesis Inhibition in apoE-Null Mice Is Dependent on the Presence of PPAR[alpha] Karen M Tordjman, Michal Vechoropoulos, Maya Ish-Shalom, Jessica Sack, Naftali Stern Tel Aviv Sourasky Medical Center, Tel Aviv, Israel Inhibition of nitric oxide synthase accelerates plaque formation in apoE-null mice, possibly by impairing the balance between angiotensin II (AII) and NO in the vessel wall. We have previously shown that PPAR[alpha]-deficient apoE-null mice are protected from atherosclerosis (1). Furthermore, we have shown that the renin angiotensin system is downregulated in mice lacking PPAR[alpha] (2). We therefore hypothesized that PPAR[alpha]-deficient apoE-null mice would be resistant to the pro-atherogenic effect caused by inhibiting NO generation with L-NAME.[br][bold]Methods:[/bold] At 4 weeks, apoE-null or apoE/PPAR[alpha]-null mice were given a sub-pressor dose of L-NAME (5 mg/l) in the drinking water or regular tap water (control). 4 weeks later, the mice were switched to a Western diet for 12 weeks, while L-NAME treatment continued throughout the experiment. Each group consisted of approximately 20 mice. Blood pressure was measured non invasively at baseline and after 16 weeks. At the end of the experiment, serum chemistry was analyzed, aortas were harvested for NADPH oxidase activity determination, and atherosclerosis was assessed from oil-red-O-stained lesions at the aortic sinus.[br][bold]Results:[/bold] As expected, lipid levels were higher in control apoE/PPAR[alpha]-null mice: cholesterol 1451[plusmn]147 vs 737[plusmn]93 mg/dl for apoE-null, P=0.0001; triglycerides 289[plusmn]48 vs 86[plusmn]6 mg/dl, P[lt]0.0001. However, L-NAME treatment increased cholesterol in the apoE-null mice to 1021[plusmn]63 mg/dl, P=0.01, but not in the apoE/PPAR[alpha]-null animals. A similar effect of L-NAME was seen for triglycerides. Aortic NADPH oxidase activity was identical in control animals 1243[plusmn]259 RLU/mg/min in apoE-null vs 1280[plusmn]159 in apoE/PPAR[alpha]-null. However, in the apoE-null mice it rose to 2587[plusmn]476 RLU/mg/min with L-NAME treatment, while it remained essentially unchanged in the apoE/PPAR[alpha]-null animals, P[lt]0.05.[br]Finally, while the extent of atherosclerosis was about 18% less pronounced in the apoE/PPAR[alpha]-null control animals, it was also completely unaffected by L-NAME treatment, whereas it went up by about 32% in the apoE-null mice. Hence, by the end of treatment it encompassed 52.7+1.7% of the sinus area in the apoE-null mice but only 31.0+2.7% in the apoE/PPAR[alpha]-null animals, P[lt]0.05. Blood pressure was unaffected by L-NAME throughout the experiment.[br][bold]Conclusions: [/bold]The data suggest that in apoE-null mice, when supply of NO is limited, the pro-oxidative and pro-atherogenic effects of angiotensin II are driven by PPAR[alpha] independently of the prevailing lipid levels.[br][br]1. Tordjman KM et al. J Clin Invest 2001; 107:1025. 2. Tordjman KM et al. Hypertension 2007; 50:945.[br][br]Sources of Research Support: A Ministry of Health Chief Scientist grant#210446 awarded to KMT; and a young investigator award from the Israeli Society for the study of Atherosclerosis to MIS.[br][br]Nothing to Disclose: KMT, MV, MI-S, JS, NS 2012-06-25T12:00:00 342ABDE 2012-06-25T00:00:00 1899-12-30T12:00:00 751 269 1884 OR22-4 OR08-01 Monday 1568 2012


1565 ENDO12L_OR22-5 ORAL SESSION: Novel Mechanisms in Lipid Metabolism (11:15 AM-12:45 PM) Testosterone Increases Hepatic Liver X Receptor and ApoE Expression and Improves Lipid Metabolism in the Testicular Feminized Mouse: A Potential Protective Mechanism Against Atherosclerosis and Fatty Liver Disease Daniel M Kelly, Samia Akhtar, Jonathan C Brooke, Vakkat Muraleedharan, Kevin S Channer, Thomas H Jones University of Sheffield, Sheffield, UK; Royal Hallamshire Hospital, Sheffield, UK; Barnsley Hospital NHS Foundation Trust, Barnsley, UK Testosterone deficiency is associated with several cardiovascular risk factors. Testosterone replacement therapy (TRT) improves insulin sensitivity, inflammation and cholesterol. Liver X receptor (LXR) is a nuclear receptor which regulates lipid and glucose metabolism, stimulates cholesterol efflux and suppresses inflammation. LXR agonists protect against atherosclerosis but cause hepatic steatosis. We have previously shown that a non-functional androgen receptor (AR) and low levels of testosterone in the testicular feminised (Tfm) mouse is associated with elevated cholesterol, hepatic steatosis and aortic fatty streak formation when fed a high-fat diet. TRT protects against hepatic steatosis and aortic lipid deposition although the exact mechanisms are unclear. We investigated whether the known protective effect of testosterone in the Tfm mouse is associated with an effect on the expression of LXR and key enzymes involved in lipid homeostasis in the liver.[br]Tfm mice fed a high-cholesterol diet for 28 weeks received either physiological TRT or placebo and were compared to wild-type littermates. Relative concentrations of liver mRNA and protein were analysed by qPCR and western blotting for the expression of LXR, Apolipoprotein E (ApoE), Hydroxyl-methyl-glutaryl CoA reductase (HMGCoA), fatty acid synthase (FAS) and acetyl coA carboxylase (ACC).[br]Compared to wild-type, Tfm mice had significantly lower mRNA expression of LXR (Relative change in mRNA Mean[plusmn]SD, 0.7[plusmn]0.09,p=0.01), ApoE (0.82[plusmn]0.1,p=0.07) and increased FAS (11.4[plusmn]4.9,p=0.05) and ACC (2.5[plusmn]0.6,p=0.05). Protein expression confirmed reduced LXR (Arbitrary Units 0.09[plusmn]0.02,p=0.05) and ApoE (0.62[plusmn]0.3,p=0.029), while FAS (1.55[plusmn]0.5,p=0.002) and ACC (2.25[plusmn]0.7,p[lt]0.001) were elevated. TRT increased LXR (1.3[plusmn]0.2,p=0.03) and ApoE (1.2[plusmn]0.12,p=0.03) mRNA in Tfm mice compared to placebo, and non-significantly reduced FAS (8.4[plusmn]2.9,p=0.11) and ACC (1.2[plusmn]0.5,p=0.1). TRT increased protein expression of LXR (0.58[plusmn]0.1,p=0.04) and ApoE (0.4[plusmn]0.1,p=0.06), while ACC (1.53vs1.22,p=0.30) and FAS (0.88vs0.89,p=0.97) decreased to wild-type levels. No effect was observed on the expression of HMGCoA.[br]TRT has a beneficial effect on hepatic expression of LXR as a master regulator of lipid metabolism and may offer atheroprotection through this mechanism. Unlike LXR agonists, TRT also decreases the expression of lipogenic enzymes and increases ApoE, thus protecting against hepatic steatosis, at least in part, via AR-independent mechanisms.[br][br]Nothing to Disclose: DMK, SA, JCB, VM, KSC, THJ 2012-06-25T12:15:00 342ABDE 2012-06-25T00:00:00 1899-12-30T12:15:00 821 269 1885 OR22-5 OR08-01 Monday 1569 2012


1566 ENDO12L_OR22-6 ORAL SESSION: Novel Mechanisms in Lipid Metabolism (11:15 AM-12:45 PM) Low Prevalence of Mutations in Known Loci for Autosomal Dominant Hypercholesterolemia in a Multi-Ethnic U.S. Cohort Zahid Ahmad, Beverley Adams-Huet, Abhimanyu Garg University of Texas Southwestern Medical Center, Dallas, TX; University of Texas Southwestern Medical Center, Dallas, TX [bold]Context:[/bold] Patients with autosomal dominant hypercholesterolemia (ADH) present with high LDL-C levels accompanied by tendon xanthomas and premature coronary heart disease (CHD). ADH is genetically heterogeneous and is caused by variants in at least three different genes: LDL receptor ([italic]LDLR[/italic]), apolipoprotein B-100 ([italic]APOB[/italic]), and proprotein convertase subtilisin-like kexin type 9 ([italic]PCSK9[/italic]).[br]Systematic screenings to identify mutations in the known genes causing ADH have been undertaken in several countries (in Europe and Japan) through nationwide screening programs and lipid clinic networks. In addition, populations such as the South African white Afrikaners, Christian Lebanese, French-Canadians, Ashkenazi Jews, and Old-Order Amish have a high prevalence of [italic]LDLR[/italic] or [italic]APOB[/italic] variants due to founder effects. Since these previous studies have been limited to ethnically/racially homogenous populations, there is a paucity of data in subjects of African origin and other minorities.[br][bold]Objective:[/bold] To examine the molecular basis of ADH in a multi-ethnic cohort from a large urban U.S. city.[br]Methods: Patients with ADH were screened for mutations in the exons and consensus splice sites of [italic]LDL[/italic]R, and in selected exons of [italic]APOB[/italic] and [italic]PCSK9[/italic]. Deletions and duplications of [italic]LDLR[/italic] exons were detected with multiplex ligation-dependent probe amplification.[br][bold]Patients:[/bold] Inclusion criteria included LDL-C [gt] 95th %tile for age and gender as well as the presence of tendon xanthomas, or a 1st degree family relative with premature CHD or LDL-C [gt] 95th %tile. All patients were screened for any secondary causes of hypercholesterolemia. A total of 38 males and 53 females, age 22 to 76 years met entry criteria and agreed to participate.[br][bold]Results[/bold]: Heterozygous variants in [italic]LDLR[/italic] were identified in 30 patients and in [italic]APOB[/italic] in one patient. The remaining 60 patients (65%) had [ldquo]unexplained ADH.[rdquo] A higher proportion of African Americans (77%) than either non-Hispanic whites (57%) or Hispanics (53%) had [ldquo]unexplained ADH.[rdquo] As compared to patients with [italic]LDLR[/italic] variants, those with [ldquo]unexplained ADH[rdquo] had lower levels of LDL-C (292 [plusmn] 47 vs 239 [plusmn] 42 mg/dL, respectively; p [lt] 0.05) and higher levels of HDL-cholesterol (45 [plusmn] 12 vs 54 [plusmn] 13 mg/dL, respectively, p [lt] 0.01).[br][bold]Conclusions:[/bold] ADH in a multi-ethnic cohort is not entirely explained by mutations in the three known genes. Our findings suggest that additional loci may contribute to ADH, especially in understudied populations such as African Americans.[br][br]Nothing to Disclose: ZA, BA-H, AG 2012-06-25T12:30:00 342ABDE 2012-06-25T00:00:00 1899-12-30T12:30:00 646 269 1886 OR22-6 OR08-01 Monday 1570 2012


1567 ENDO12L_OR23-1 ORAL SESSION: Mechanisms Linking Peripheral Metabolism to Central Regulation of Appetite (11:15 AM-12:45 PM) POMC-GLP-2R Signaling and Action in the Control of Feeding Behavior and Gastric Motility Xinfu Guan, Xuemei Shi, Depei Li, Xiaojie Li, Yi Wang, Benny Chang, Lawrence Chan Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; University of Texas MD Anderson Cancer Center, Houston, TX Glucagon-like peptides (GLP-1/2) are released in response to food intake. Central infusion of GLP-2 suppresses food intake and gastric motility, and this is supposedly mediated through its receptor (GLP-2R) action in the brain. However, the physiological relevance of GLP-2R in the brain is unknown. Thus, our aims were to determine if GLP-2R in POMC neurons regulates feeding behavior and gastric motility. [1] Using the Cre-LoxP system, we recently generated mice that lack [italic]Glp2r[/italic] expression in POMC neurons. Mice with [italic]Glp2r[/italic] deletion in POMC neurons showed late-onset obesity, higher food intake, and increased meal frequency. Importantly, the KO mice also showed accelerated gastric emptying using [sup]13[/sup]C-octanoic acid breath test. For liquid meal, half-excretion time was 29.0 [plusmn] 4.5 and 51.2 [plusmn] 4.5 min; and lag phase 17.8 [plusmn] 3.0 and 31.5 [plusmn] 3.0 min for the KO and WT mice, respectively. Notably, proglucagon expression increased in the KO brain, pointing to a negative relationship between GLP-2 and GLP-2R in the brain. [2] We generated POMC-specific [italic]p110[alpha][/italic] knockout mice and POMC-specific FoxO1-GFP over-expression mice to further define cellular mechanisms underlying GLP-2-initiated anorexic action. Using immunoprecipitation, we found that GLP-2R interacted with p85[alpha] (a regulatory subunit of PI3K) in both transfected HEK293 cells and cultured hippocampal neurons. GLP-2 acutely induced GLP-2R translocation from membrane to cytoplasm, and enhanced Akt phosphorylation on Ser473 in a PI3K-dependent manner. Moreover, GLP-2 (at 20 nM for 30 min) induced nuclear exclusion of FoxO1-GFP in POMC neurons, and this effect was abolished in POMC-specific p110[alpha] (a catalytic subunit of PI3K) KO mice, suggesting that GLP-2 may disinhibit expression of FoxO1-targeted genes (such as [italic]Pomc [/italic]and [italic]Cpe[/italic]) in POMC neurons. Furthermore, GLP-2 differentially modulated excitation of functionally-segregated POMC neurons in a PI3K-dependent manner. In conclusion, [italic]Glp2r [/italic]deletion in POMC neurons impairs feeding behavior and gastric motility; and GLP-2 acutely activates the PI3K signaling pathway in POMC neurons to modulate neuronal excitation and gene expression. Therefore, CNS GLP-2R plays an important, physiological role in the control of food intake and gastric emptying.[br][br]Sources of Research Support: NIH Grant DK075489 and DK084125 awarded to XG; USDA Grant 6250-51000-043 awarded to XG.[br][br]Nothing to Disclose: XG, XS, DL, XL, YW, BC, LC 2012-06-25T11:15:00 351 2012-06-25T00:00:00 1899-12-30T11:15:00 987 270 1887 OR23-1 OR27-02 Monday 1571 2012


1568 ENDO12L_OR23-2 ORAL SESSION: Mechanisms Linking Peripheral Metabolism to Central Regulation of Appetite (11:15 AM-12:45 PM) Glutamate Release from Leptin Receptor-Expressing Neurons Is Required for Normal Body Weight Regulation Yuanzhong Xu, Martin G Myers, Qingchun Tong University of Texas Medical School at Houston, Houston, TX; University of Michigan, Ann Arbor, MI Leptin acts on brain neurons to maintain body weight homeostasis. Brain neurons expressing leptin receptors (LepR neurons) are mainly located in the hypothalamus, but also, to a much lesser degree, in a few other brain sites. The major brain areas with abundant LepR neurons show high levels of expression of either vesicular GABA transporter or vesicular glutamate transporter 2 (Vglut2), suggesting that the majority of LepR neurons are either GABAergic or glutamatergic. Indeed, previous studies have demonstrated an important role for the leptin-LepR neurons-GABA neural pathway in mediating leptin action on body weight regulation. However, the extent to which glutamate release from LepR neurons mediates leptin action on body weight is unknown. To directly address this, we generated mice with disruption of glutamate release from LepR neurons by deletion of Vglut2 specifically in these neurons. Mice with deletion of Vglut2 in LepR neurons developed obesity on chow diet, which was associated with reduced energy expenditure (O2 consumption) and respiratory exchange ratio. Despite normal food intake on chow diet, these mice displayed a blunt response in food intake to acute leptin administration. Taken together, these results demonstrate that glutamate release from LepR-expressing neurons is required for normal body weight homeostasis.[br][br]Sources of Research Support: American Heart Association and NIH grant R56DK089426 to QT.[br][br]Nothing to Disclose: YX, MGM, QT 2012-06-25T11:30:00 351 2012-06-25T00:00:00 1899-12-30T11:30:00 1647 270 1888 OR23-2 OR27-02 Monday 1572 2012


1569 ENDO12L_OR23-3 ORAL SESSION: Mechanisms Linking Peripheral Metabolism to Central Regulation of Appetite (11:15 AM-12:45 PM) Estradiol Increases the Central Effects of Brain-Derived Neurotrophic Factor Haifei Shi, Zheng Zhu, Shiva PD Senthil Kumar, Xian Liu Miami University, Oxford, OH Estrogens suppress food intake and decrease body weight. Compelling evidence suggests that estradiol (E2), the primary form of estrogen, reduces caloric intake by enhancing the strength of other anorexigenic signals. Central administration brain-derived neurotrophic factor (BDNF) induces a negative energy balance by decreasing caloric intake. We previously found that female rats at estrous stage, a phase following the peak of E2 being secreted and animals[apos] behavior is mostly influenced by the endogenous estrogens, responded to a lower dose of BDNF to suppress feeding and induce weight loss than males and females at other stages of phase, indicating that the anorexigenic effect of BDNF is influenced by sex and stage of estrous cycle. The first goal of this study is to determine whether E2 modulates the anorectic effects of BDNF. Adult female Long-Evans rats received intra-third-cerebroventricular (i3vt) cannulation and ovariectomy (OVX). One group of OVX rats received a cyclic regimen of E2 replacement (one subcutaneous injection every four days) at a physiological dose to mimic the normal changes in plasma E2 levels across the estrous cycle. The other group of OVX rats received vehicle (oil) injections. BDNF at a low dose (0.1 [micro]g) significantly reduced food intake in E2-treated rats but not in oil-treated rats, suggesting that the minimally effective anorectic dose of BDNF is lower after cyclic E2 replacement. Although a larger dose of BDNF (0.3 [micro]g) decreased food intake in oil-treated OVX rats, this anorectic effect was significantly less than that in E2-treated OVX rats. These findings suggest that a physiological dose of E2 is sufficient to increase BDNF-induced reduction of feeding. The second goal of this study is to determine whether BDNF gene expression is regulated by endogenous estrogens of cycling rats and is regulated by E2 using OVX rats. Estrous female rats, but not rats at other estrous stages, had significantly higher VMH BDNF mRNA levels than male rats. Oil-treated OVX rats had significantly lower VMH BDNF mRNA, compared with E2-treated females. This alteration induced by OVX was almost completely restored by cyclic replacement with E2, indicating that E2 regulates BDNF gene expression in the VMH. We conclude that E2 regulates BDNF gene expression and enhances the anabolic effects of BDNF in OVX rats. These data indicate that an increased signaling of endogenous BDNF may mediate estrogen-induced inhibitory effect on feeding.[br][br]Sources of Research Support: National Institutes of Health National Institute of Diabetes and Digestive and Kidney Diseases [Grant 1R15DK090823-01 to HS] and American Heart Association [Grant 10SDG4520028 to HS].[br][br]Nothing to Disclose: HS, ZZ, SPDSK, XL 2012-06-25T11:45:00 351 2012-06-25T00:00:00 1899-12-30T11:45:00 1029 270 1889 OR23-3 OR27-02 Monday 1573 2012


1570 ENDO12L_OR23-4 ORAL SESSION: Mechanisms Linking Peripheral Metabolism to Central Regulation of Appetite (11:15 AM-12:45 PM) Grazing Induces Hyperghrelinemia and Increases Fat Storage Efficiency in Rats Thomas W Tilston, Bradley Arms-Williams, Daniel J Ball, Lyndsey Phelps, Jeffrey S Davies, Timothy Wells Cardiff University, Cardiff, UK; Swansea University, Swansea, UK The current obesity epidemic has arisen in the context of a shift from regular meal consumption towards grazing or snacking behaviour (1,2) and it is estimated that this increase in eating occasions is associated with a rise in energy intake of 39kcal/day (2). While an escalating energy intake may contribute to the development of obesity, our understanding of the physiological impact of feeding frequency has been hampered by an inability to impose consistent feeding patterns in laboratory animals.[br]We have used a CLAMS-based system to investigate the effect of 3 weeks of grazing (consumption of 1/24th of the total daily food intake of [italic] ad libitum[/italic]-fed controls every 30 mins during the dark phase (18.00h-06.00h)) and meal feeding (three 1-hour periods of [italic] ad libitum [/italic] food access at 18.00h, 23.30h and 05.00h) on growth and adiposity in rats, using automated serial blood sampling and LH-PCR and qPCR to assess hormone and gut microbiota profiles.[br]Grazing rats consumed only 84% of their total daily allowance (p[lt]0.001). While this was accompanied by a mild (2%) reduction in skeletal growth, body weight was unaffected; lipid storage efficiency (retroperitoneal fat mass/caloric intake) being augmented by 27% (p[lt]0.05). Conversely, although meal-fed rats showed similar reductions in food intake and skeletal growth, body weight gain decelerated in parallel with food intake. In grazing rats the usual decline in circulating ghrelin during the late light phase was replaced by a sustained surge in ghrelin secretion, with a gradual rise in plasma ghrelin during the dark (feeding) phase. Although total growth hormone (GH) secretion was unaffected, the daily nadir in GH peaks was shifted into the early dark phase. Corticosterone profiles were unaffected. In addition, grazing reduced [italic] bacteriodetes [/italic] in fecal pellets (p[lt]0.05).[br]Thus, the pattern of feeding profoundly influences lipid metabolism. Grazing modifies the gut microbiota profile to promote absorption of short-chain fatty acids (3), augments the secretion of ghrelin to enhance fat storage and aligns the daily nadir in the lipolytic influence of GH with the availability of circulating nutrients. When combined with access to high-energy diets, the obesogenic impact of this pattern of feeding may be a significant factor in the current obesity epidemic.[br][br](1) Rowbotham J, Clayton P. 2008; J R Soc Med 101:454. (2) Duffey KJ, Popkin BM. 2011; PLoS Med 8:e1001050. (3) Greiner T, B[auml]ckhed F. 2011; Trends Endocrinol Metab 22:117.[br][br]Sources of Research Support: Waterloo Foundation Grant 267-495; Cardiff University School of Biosciences Equipment Grant awarded to TW.[br][br]Nothing to Disclose: TWT, BA-W, DJB, LP, JSD, TW 2012-06-25T12:00:00 351 2012-06-25T00:00:00 1899-12-30T12:00:00 607 270 1890 OR23-4 OR27-02 Monday 1574 2012


1571 ENDO12L_OR23-5 ORAL SESSION: Mechanisms Linking Peripheral Metabolism to Central Regulation of Appetite (11:15 AM-12:45 PM) Fructose Compared to Glucose Ingestion Preferentially Activates Brain Reward Regions in Response to High-Calorie Food Cues in Young, Obese Hispanic Females Kathleen Alanna Page, Shan Luo, Ana Romero, Tanja Adam, Houchun Harry Hu, John Monterosso University of Southern California Keck School of Medicine, Los Angeles, CA; University of Southern California, Los Angeles, CA; University of Southern California Keck School of Medicine, Los Angeles, CA; University of Southern California/Children[apos]s Hospital Los Angeles, Los Angeles, CA Increases in dietary fructose consumption parallel the rising rates in obesity. Animal data show that the central administration of glucose reduces food intake, whereas fructose administration promotes food intake. We tested the hypothesis that fructose compared to glucose ingestion would result in greater hypothalamic and brain reward center activation and increased hunger and desire for food in response to high-calorie (HC) food cues in young, obese Hispanic females, a population at high risk for continued weight gain.[br]A 3-Tesla scanner was used to perform functional magnetic resonance imaging (fMRI) on 13 young (23[plusmn]2 years), obese (34[plusmn]4 kg/m[sup]2[/sup]) Hispanic females. Participants underwent 2 fMRI sessions together with ingestion of either fructose or glucose (50g in 150 mL water) in a double blinded, random-order crossover design. Scanning was performed as subjects viewed images of HC and low-calorie foods and non-food (NF) items using a block design. After each block participants rated hunger and desire for sweet and savory foods. Total scan time was [sim]30 min. Drinks were ingested after 15 min of baseline scanning. Brain activation to HC foods was determined by calculating a contrast of HC minus NF images. Effects of fructose vs. glucose ingestion on brain signal change to HC food cues was examined by comparing the contrast maps after fructose vs. glucose ingestion. Ratings of hunger and desire for food during HC vs. NF blocks were compared with paired t-tests.[br]Independent of sugar ingestion, viewing HC food images activated the hypothalamus, striatum, insula and orbitofrontal cortex (OFC) (Z[gt]2.3, p[lt]0.05,corrected) and increased hunger ([italic]t[/italic](1,12)=5.317,p=0.0001) and desire for sweet ([italic]t[/italic](1,12)=2.942,p=0.012) and savory foods ([italic]t[/italic](1,12)=4.221,p=0.001). Hunger ([italic]t[/italic](1,12)=4.441, p=0.001) and desire for savory foods (t(1, 12)=2.735,p=0.018) were greater after drink ingestion regardless of type of sugar. Whole-brain analysis suggested that fructose compared to glucose ingestion produced greater activation in the nucleus accumbens and OFC, brain regions involved in reward processing, in response to HC food cues (Z[gt]1.6,uncorrected).These data suggest that viewing HC food images activates brain reward regions and increases hunger and desire for sweet and savory foods, and that fructose compared to glucose ingestion preferentially activates brain reward regions in young, obese Hispanic women. These neural and behavioral responses could promote food intake and weight gain.[br][br]Sources of Research Support: National Center on Minority Health and Health Disparities P60 MD002254-01 Pilot Project awarded to KAP; NIDDK 1K23DK092702-01 awarded to KAP.[br][br]Nothing to Disclose: KAP, SL, AR, TA, HHH, JM 2012-06-25T12:15:00 351 2012-06-25T00:00:00 1899-12-30T12:15:00 1666 270 1891 OR23-5 OR27-02 Monday 1575 2012


1572 ENDO12L_OR23-6 ORAL SESSION: Mechanisms Linking Peripheral Metabolism to Central Regulation of Appetite (11:15 AM-12:45 PM) Oxytocin Secretion and Food Motivation Neurocircuitry in Anorexia Nervosa Elizabeth A Lawson, Laura M Holsen, McKale Santin, Erinne Meenaghan, Anne E Becker, David B Herzog, Jill M Goldstein, Anne Klibanski Massachusetts General Hospital, Harvard Medical School, Boston, MA; Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA; Massachusetts General Hospital, Harvard Medical School, Boston, MA Animal data suggest that the neuropeptide oxytocin (OT) is a satiety hormone. We have shown that anorexia nervosa (AN), a disorder characterized by food restriction and severe weight loss, is associated with low nocturnal OT secretion. Using functional magnetic resonance imaging (fMRI), we reported hypoactivation of brain regions involved in food motivation, including the hypothalamus (control center for appetitive signals), amygdala (important for learning satiety cues and assessing reward value), hippocampus (implicated in processing food-related memories), orbitofrontal cortex (OFC, integrates emotion and reward expectation) and insula (houses the primary taste cortex, integrates visceral signals, and modulates motivational behavior), in AN vs. normal-weight controls (HC). We hypothesized that abnormal OT secretion would be associated with severity of disordered eating psychopathology and hypoactivation of food motivation neural pathways. We studied 35 women: 13 AN, 9 weight-recovered AN (ANWR) and 13 HC of comparable age. Serum OT and leptin levels were measured fasting and 30, 60, and 120 min after a 400kcal mixed meal. The Eating Disorder Examination-Questionnaire (EDE-Q) was used to assess disordered eating psychopathology. fMRI was performed during visual processing of food vs. non-food stimuli pre- and post-meal. We found that mean OT levels were higher in AN than HC at 60 and 120 min, and lower in ANWR than HC at 0, 30 and 120 min and AN at all timepoints pre- and post-meal (p[lt]0.05). Mean OT area under the curve (AUC) was highest in AN, intermediate in HC, and lowest in ANWR. In AN and ANWR, OT AUC was associated with the severity of disordered eating psychopathology (r=0.76, p[lt]0.0001), independent of leptin AUC. OT secretion accounted for a substantial proportion of between-group variance in fMRI activation: pre-meal, HC[gt]AN hypothalamus (22%), amygdala (42%), hippocampus (13%) and OFC (21%); post-meal HC[gt]AN amygdala (22%) and insula (9%); pre-meal HC[gt]ANWR insula (20%); post-meal, ANWR[gt]AN insula (13%). These data indicate that OT secretion is associated with hypoactivation of food motivation circuitry in active (a [ldquo]state[rdquo] characteristic) and weight-recovered (a [ldquo]trait[rdquo] characteristic) AN. Importantly, the insula integrates interoceptive signaling, and a lack of interoceptive awareness is a defining feature of AN. Inherent abnormalities in OT secretion may therefore contribute to underlying deficits that increase susceptibility to AN.[br][br]Sources of Research Support: Harvard Catalyst | The Harvard Clinical and Translational Science Center (NIH Award #UL1 RR025758); Harvard K12 HD051959 Building Interdisciplinary Research Careers in Women[apos]s Health (BIRCWH) Program supported by National Institutes of Health Office of Research in Women[apos]s Health (ORWH).[br][br]Nothing to Disclose: EAL, LMH, MS, EM, AEB, DBH, JMG, AK 2012-06-25T12:30:00 351 2012-06-25T00:00:00 1899-12-30T12:30:00 922 270 1892 OR23-6 OR27-02 Monday 1576 2012


1573 ENDO12L_OR24-1 ORAL SESSION: Coactivators [amp] Corepressors: Regulators of Gene Expression [amp] Physiology (11:15 AM-12:45 PM) [italic]ZNF764[/italic] Haploinsufficiency May Explain Multiple Steroid Hormone Resistance Associated with 16p11.2 Microdeletion Tomoshige Kino, Maria G Pavlatou, Andreas Moraitis, Robin L Nemery, Margarita Raygada, Constantine A Stratakis National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD; National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD; Joe DiMaggio Children[apos]s Hospital, Hollywood, FL Nuclear hormone receptors (NRs) exert their transcriptional effects though shared cofactor molecules, thus, defects in such intermediate proteins may be associated with multiple hormone resistance. Microdeletion of small chromosomal segments results in hereditary or sporadic diseases by affecting expression of residing genes. We describe a 7-year boy with mild resistance to glucocorticoids, thyroid hormones, and possibly, androgens. He had elevated levels of morning serum cortisol (34.5 ng/dl, normal range (nr): 6-23) and plasma ACTH (122 pg/ml, nr: 9-52), which were not suppressed by dexamethasone. Serum TSH concentration was elevated: 6.88 mIU/L (nr: 0.4-4.0), while that of free T4 was in a normal range: 1.9 ng/dl (nr: 0.8-2.3). He had significant dolichocephaly, hypertelorism, micropenis and small, underdeveloped scrotum. He was also diagnosed as being in the autism spectrum disorder, and had developmental delay and several facial morphologic manifestations. We found with aCGH and FISH analyses as well as subsequent real-time PCR a [sim]1.1 Mb size heterozygotic 16p11.2 microdeletion, which had an additional [sim]0.5 Mb extension to a common [sim]600 Kb 16p11.2 microdeletion known to be associated with autism, developmental delay and characteristic facial manifestations. The siRNA-based screening employing 38 siRNAs for the genes located in the deleted segment revealed that knockdown of the zinc finger protein 764 (ZNF764), which is located in the deleted segment unique to our case, significantly reduced glucocorticoid-, androgen- and thyroid hormone-induced transcriptional activity of their corresponding responsive genes in HeLa cells, while its overexpression enhanced their transcriptional activity. The activities of the estrogen receptor, CREB and p53 were not affected in these cells. Protein/mRNA levels of ZNF764 were reduced in patient[apos]s peripheral blood mononuclear cells, while those of GR, TR and AR were maintained. Exogenously supplemented ZNF764 recovered responsiveness to glucocorticoids in the patient[apos]s Epstein-Barr virus-transformed lymphocytes. The effect of ZNF764 on GR transcriptional activity was mediated through cooperation with a general NR coactivator, transcriptional intermediary factor 1. Taken together, these results indicate that [italic]ZNF764[/italic] haploinsufficiency caused by microdeletion may be responsible for the multiple hormone resistance observed in our patient. ZNF764 appears to be involved in glucocorticoid, androgen and thyroid hormone action.[br][br]Sources of Research Support: This study was funded by the [italic]Eunice Kennedy Shriver[/italic] National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD.[br][br]Nothing to Disclose: TK, MGP, AM, RLN, MR, CAS 2012-06-25T11:15:00 371 2012-06-25T00:00:00 1899-12-30T11:15:00 178 271 1893 OR24-1 OR23-01 Monday 1577 2012


1574 ENDO12L_OR24-2 ORAL SESSION: Coactivators [amp] Corepressors: Regulators of Gene Expression [amp] Physiology (11:15 AM-12:45 PM) Inhibition of PELP1 Oncogenic Functions with Cell-Penetrating Peptides Monica Mann, Ratna K Vadlamudi University of Texas Health Science Center at San Antonio, San Antonio, TX; University of Texas Health Science Center at San Antonio, San Antonio, TX Estrogen receptor alpha (ER) is the fundamental transcription factor in breast cancer progression. Therapies targeting ER are effective; however, many patients ultimately develop resistance and recur as metastases. ER signaling is mediated by coregulator proteins that can have altered expression or function that promotes therapy resistance and metastasis by enhancing hormone-independent ER signaling and by epigenetic changes. An alternative strategy for inhibiting ER signaling in therapy resistant and metastatic tumors is to inhibit the function of ER coregulators. PELP1 is an ER coactivator and proto-oncogene which is overexpressed in breast cancer, promotes breast cancer progression by epigenetic changes at ER target genes and is an independent marker of poor prognosis. The objective of this study is to develop an inhibitor of PELP1-mediated ER coactivator functions. Since PELP1 lacks any known enzymatic activity, we targeted PELP1 protein-protein interactions to inhibit its oncogenic functions. We used a yeast two-hybrid system to screen a library of random peptides and identified 23 peptides with high affinity for PELP1. Two of these PELP1-inhibiting peptides (PIP-1 and PIP-2) significantly block PELP1 coactivator functions. We used a TAT-peptide fusion of PIPs to deliver the peptides into cells, confirmed cellular uptake of the PIPs by fluorescent microscopy and confirmed PELP1-PIP binding by peptide pull-down assays. Both PIP- 1 and -2 significantly inhibited PELP1-mediated proliferation in two ER-positive breast cancer cells. A GenBank search revealed homology of PIPs with the histone methyltransferase G9a/KMT1C that is overexpressed in aggressive breast cancer, contributes to epigenetic silencing of tumor suppressors and promotes cancer invasion and metastasis. [italic] In vivo [/italic] co-immunoprecipitation and an [italic] in vitro [/italic] binding assay confirmed PELP1 interaction with G9a, and peptide competition assays indicated that the PIPs could interrupt PELP1-G9a interaction. Both PIPs substantially inhibited PELP1-mediated cell invasion in Boyden chamber assays. Interestingly, disrupting PELP1-G9a interaction with PIPs sensitized resistant cells to hormonal therapy. Mechanistic studies revealed that PELP1-G9a interactions play a critical role in epigenetic changes in therapy resistant cells. In conclusion, our studies identified novel inhibitors of PELP1 that can be used for therapeutic targeting of ER-coregulator driven cancer progression and therapy resistance.[br][br]Sources of Research Support: NIH Grant CA095681; CPRIT pre-doctoral fellowship RP101491.[br][br]Nothing to Disclose: MM, RKV 2012-06-25T11:30:00 371 2012-06-25T00:00:00 1899-12-30T11:30:00 1702 271 1894 OR24-2 OR23-01 Monday 1578 2012


1575 ENDO12L_OR24-3 ORAL SESSION: Coactivators [amp] Corepressors: Regulators of Gene Expression [amp] Physiology (11:15 AM-12:45 PM) The Nuclear Receptor Corepressor SMRT Coactivates p53 Transcription Function and Modulates Cellular Response to DNA Damage Anbu Karani Adikesavan, Sudipan Karmakar, Patricia Pardo, Liguo Wang, Wei Li, Carolyn L Smith Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX The corepressor SMRT represses the transcriptional activity of unliganded nuclear receptors and plays a key role in regulating the expression of genes involved in growth, development and metabolism through recruitment of histone deacetylases such as HDAC3 and compaction of chromatin. In order to profile gene expression regulated by SMRT, MCF-7 breast cancer cells were transfected with control or SMRT-specific siRNA. Affymetrix GeneChip microarray analyses of isolated RNA followed by gene ontological analyses revealed that SMRT depletion was associated with a decrease in the expression of genes involved in DNA damage response such as BRCA1 and CHEK1. Consistent with this, levels of the DNA damage marker [gamma]H2AX and PARP cleavage, a marker of apoptosis, were elevated in SMRT-depleted [italic]versus[/italic] control cells treated with doxorubicin indicating a role for this coregulator in mediating cellular responses to DNA damaging agents. Several of the DNA damage response genes identified by microarray are p53 targets, suggesting a role for SMRT in regulation of p53 transcriptional activity. Silencing SMRT expression decreased luciferase expression from two distinct p53-dependent reporter genes, without affecting p53 protein expression. In addition, doxorubicin-induced expression of the p53 target gene, p21 was reduced in SMRT-depleted [italic]versus[/italic] control cells. In contrast, overexpression of SMRT increased the expression of the p21-Luc reporter gene. Chromatin immunoprecipitation assays detect doxorubicin-induced recruitment of SMRT to p53 binding sites in the p21 promoter. Co-immunoprecipitation experiments demonstrate SMRT [italic]in vivo[/italic] interaction with endogenous p53 in both HCT116 colon and C4-12 breast cancer cell lines, while GST pull-down assays reveal a direct interaction of p53 with the N-terminal regions of SMRT[alpha] and the SMRT[beta] splice variant. Mapping experiments conducted by GST pull-down assay revealed that p53 binds to a region encompassing the deacetylase activation domain (DAD) of SMRT which mediates HDAC3 binding and activation of its enzymatic activity. Collectively, these data highlight a biological role for SMRT in mediating DNA damage responses by promoting p53 transcriptional activity, and suggest a model where p53 binding to the region encompassing the DAD displaces HDAC3 from SMRT resulting in loss of repression, and coactivation of p53-dependent gene expression.[br][br]Sources of Research Support: NIH grant DK53002 to CLS.[br][br]Nothing to Disclose: AKA, SK, PP, LW, WL, CLS 2012-06-25T11:45:00 371 2012-06-25T00:00:00 1899-12-30T11:45:00 1880 271 1895 OR24-3 OR23-01 Monday 1579 2012


1576 ENDO12L_OR24-4 ORAL SESSION: Coactivators [amp] Corepressors: Regulators of Gene Expression [amp] Physiology (11:15 AM-12:45 PM) Loss of SLIRP, a Nuclear Receptor Corepressor, Produces an Advantageous Metabolic Phenotype Shane M Colley, Larissa Wintle, Vance Matthews, Richelle Searles, Michael Phillips, Peter J Leedman Western Australian Institute for Medical Research, Perth, Australia; Royal Perth Hospital, Perth, Australia; University of Western Australia, Perth, Australia Nuclear receptors (NRs) and their signaling pathways play central roles in the regulation of glucose and lipid metabolism and are important therapeutic targets. Previously we discovered SLIRP, a Steroid receptor RNA Activator (SRA)-binding NR coregulator, which is a potent repressor of NR transcriptional activity (1). In particular, SLIRP represses PPAR activity in vitro in insulin sensitive muscle and adipose cell lines. Intriguingly, SLIRP is highly expressed in mitochondria-rich tissues (muscles, heart, liver, testis) (1), the majority of SLIRP is mitochondrial and it has been shown to regulate mitochondrial gene expression (2). For these reasons, we explored the functional role of SLIRP on metabolism in vivo using the SLIRP knock-out mouse (SLIRPko). SLIRPko mice are more resistant to diet induced diabetes than wild type SLIRP (SLIRPwt) animals. In particular, SLIRPko mice are leaner than wild type animals, they have enhanced glucose tolerance (via Glucose Tolerance Testing, GTT) as well as increased insulin sensitivity (via Insulin Tolerance Testing, ITT). Comparison of genes expressed in SLIRPko and wt mouse muscle highlighted the differential expression of a number of key regulators of metabolism including Egr1 and Retsat (a PPAR target gene). Egr1 mRNA expression was significantly downregulated in SLIRPko muscle, consistent with its role as a regulator of insulin sensitivity. In contrast, Retsat mRNA expression was increased in the SLIRPko mice, consistent with its role as a regulator of adipogenesis, the leaner phenotype we observed in the SLIRPko mice and the predicted loss of PPAR repression in the SLIRPko animals. Taken together, these data support the concept that SLIRP is a regulator of key metabolic genes that can impact on body weight, glucose homeostasis and insulin sensitivity. In addition, as the SLIRPko mouse results in an advantageous metabolic phenotype, the targeting of SLIRP may provide new therapeutic opportunities in diabesity.[br][br]1. Hatchell et al. Mol Cell 2006; 22:657. 2. Sarasman et al. Mol Biol Cell 2010; 21:1313.[br][br]Sources of Research Support: The NHMRC and DART.[br][br]Nothing to Disclose: SMC, LW, VM, RS, MP, PJL 2012-06-25T12:00:00 371 2012-06-25T00:00:00 1899-12-30T12:00:00 1148 271 1896 OR24-4 OR23-01 Monday 1580 2012


1577 ENDO12L_OR24-5 ORAL SESSION: Coactivators [amp] Corepressors: Regulators of Gene Expression [amp] Physiology (11:15 AM-12:45 PM) SRC-2 Has Dual Function in Hepatic Glucose Metabolism Bin Zhang, Jean-Francois Louet, Subhamoy Dasgupta, Bert W O[apos]Malley Baylor College of Medicine, Houston, TX Steroid receptor coactivator-2 (SRC-2), a member of the p160 family of hormone nuclear receptor coregulators, has been reported to be a [apos]master regulator[apos] to control energy metabolism in different tissues or organs. Particularly, SRC-2 is a critical regulator of hepatic glucose production by regulating glucose-6-phosphatase (G6Pase) expression during the fasting stage. In our current study, SRC-2 was found to regulate hepatic glycolysis as well as gluconeogenesis. After re-feeding, SRC-2 knockout mice showed significantly higher blood glucose levels and less hepatic glycogen storages compared with WT mice. Oppositely, in the fasting stage, hypoglycemia and higher levels of glycogen storage have been observed in SRC-2 knockout mice. On the basis of these results, we postulated that the dual contribution of SRC-2 to hepatic glycolysis and gluconeogenesis may be due to its transcriptional coregulator functions in different genes. Indeed, we screened a series of rate-limiting glycolytic enzymes and found the hepatic levels of glucokinase (GCK) were significantly decreased in the SRC-2 knockout mice. The deficiency of GCK is closely related to insulin resistance and diabetes, and the mutations in GCK lead to maturity onset diabetes of the young, type 2 (MODY2). As a coactivator, SRC-2 was found to regulate GCK mRNA transcription in hepatocyte cells, and the binding ability of SRC-2 on liver GCK promoter was also validated by ChIP assay. Our most recent evidence indicates that SRC-2 contributes mostly to G6Pase in fasting, and switches to regulate GCK during re-feeding. The ability of SRC-2 to either regulate GCK or G6Pase depends on different kinase pathways, which was proved by using different chemical inhibitor assays. For example, PI3K/AKT inhibitors (Wortmannin) only blocked the SRC-2 induced GCK overexpression but not G6Pase. In contrast, PKA inhibitors (H-98) decreased G6Pase induced by SRC-2 but had no effect on GCK. Altogether, our results suggest that [apos]phosphorylation codes[apos] on SRC-2 may play an important role in controlling functional switch between glycolysis and gluconeogenesis. Further identification of the [apos]codes[apos] and the related kinase regulators may contribute to the potential therapeutic study for maintaining glucose homeostasis.[br][br]Nothing to Disclose: BZ, J-FL, SD, BWO 2012-06-25T12:15:00 371 2012-06-25T00:00:00 1899-12-30T12:15:00 1873 271 1897 OR24-5 OR23-01 Monday 1581 2012


1578 ENDO12L_OR24-6 ORAL SESSION: Coactivators [amp] Corepressors: Regulators of Gene Expression [amp] Physiology (11:15 AM-12:45 PM) Regulation of Steroid Receptor Coactivator (SRC)-3 Expression and Function by Mutant G Proteins in Cancer Sue Anne Chew, Bin He, Vijay Kumar Eedunuri, Diego Jacinto Bedoya, Chuandong Geng, Ashesh Shah, Vassiliki Poulaki, David Lonard, Bert W O[apos]Malley, Nicholas Mitsiades Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, LA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, LA; Veterans Affairs Boston Healthcare System, Jamaica Plain/Boston, MA [bold]Background:[/bold] The Steroid Receptor Coactivator (SRC)-3 (AIB1/NCoA3) is a key pleiotropic [ldquo]master regulator[rdquo] of transcription factors necessary for cancer cell proliferation, survival, metabolism and metastasis. SRC-3 overexpression and overactivation occurs in numerous human cancers, due to a variety of genomic, transcriptional and post-translational mechanisms, and is associated with poor clinical outcomes and resistance to therapy, suggesting that it is an important therapeutic target. We have previously reported that protein kinase C (PKC) family members can phosphorylate SRC-3 and prevent its proteasome-mediated degradation. We also found that somatic mutations in the guanine nucleotide-binding protein (G protein) [alpha] subunits G[alpha]q and G[alpha]11 (encoded by GNAQ and GNA11, respectively), such as those found in most uveal melanomas (UMs), drive UM cell growth and survival via a pathway involving phospholipase C[beta] (PLC[beta]), PKC and protein kinase D (PKD).[br][bold]Methods:[/bold] Using RNA interference (RNAi), small molecule inhibitors (SMIs) and adenoviral-mediated gene transfer, we explored the regulation of SRC-3 expression and function by mutant (mt) G[alpha] proteins in UM cell lines.[br][bold]Results:[/bold] Depletion of G[alpha]q or PKC[alpha] or PKD1 (by RNAi), as well as PKC SMIs, promoted the proteasomal degradation of SRC-3 and induced, in GNAQ-mt cells, caspase-independent cell death with hallmarks of autophagy. SRC-3 siRNA potently induced cell death in GNAQ-mt, but not G[alpha]-wild type UM cells. Exogenous overexpression of SRC-3 protected GNAQ[ndash]mt cells from cell death induced by GNAQ siRNA or PKC inhibition. Gene expression profiling of GNAQ-mt cells treated with GNAQ siRNA revealed regulation of transcripts involved in cell signaling and migration, nutrient transport and metabolism. Direct recruitment of SRC-3 to respective gene promoters was confirmed by chromatin immunoprecipitation-PCR analysis. In agreement with the downregulation of cell membrane transporters for select nutrients (e.g. monocarboxylates and several aminoacids), inhibition of the G[alpha]q/PKC/SRC3 pathway suppressed the capacity of GNAQ-mt cells to use the corresponding nutrients as energy source, attenuated oxygen consumption and led to autophagy.[br][bold]Conclusions:[/bold] G[alpha]-induced signaling promotes the post-translational stabilization of SRC-3 in a PKC- and PKD-dependent manner. G[alpha]-mt UM cells are exquisitely dependent on SRC-3 for metabolism, proliferation and survival, highlighting SRC-3 as a potential effector of G[alpha]-induced oncogenic signaling.[br][br]Sources of Research Support: The authors acknowledge the joint participation by Adrienne Helis Malvin Medical Research Foundation through its direct engagement in the continuous active conduct of medical research in conjunction with Baylor College of Medicine. B.H. is supported by NIH/NIDDK grant 5K01DK081446. N.M. is a Dan L. Duncan Scholar and a Caroline Wiess Law Scholar at Baylor College of Medicine and his work is also supported by the Conquer Cancer Foundation of the American Society of Clinical Oncology (ASCO) Career Development Award and by the Prostate Cancer Foundation.[br][br]Nothing to Disclose: SAC, BH, VKE, DJB, CG, AS, VP, DL, BWO, NM 2012-06-25T12:30:00 371 2012-06-25T00:00:00 1899-12-30T12:30:00 1664 271 1898 OR24-6 OR23-01 Monday 1582 2012


1579 ENDO12L_OR25-1 ORAL SESSION: Female Reproduction: Gonadotropin Control [amp] Metabolic Responses (11:15 AM-12:45 PM) A Novel Reproductive Hormone, Phoenixin Gina LC Yosten, Rong-Ming Lyu, Jaw-Kang Chang, Chloe W Bryant, Nae J Dun, Siok Le Dun, Aaron Hsueh, Willis K Samson Saint Louis University School of Medicine, Saint Louis, MO; Phoenix Pharmaceuticals, Burlingame, CA; Temple University, Philadelphia, PA; Stanford University, Palo Alto, CA Based on bioinformatic analyses of conserved sequences and dibasic cleavage sites, a previously undescribed peptide was identified and isolated from rat hypothalamus using an antibody raised against the synthetic peptide. Mass spectrometry revealed that endogenous peptide, phoenixin, existed both as a 14- and 20-amino acid peptide that was C-terminally amidated. Using an EIA, we detected phoenixin in various central and peripheral tissues, including hypothalamus, pituitary, thymus, heart, lung, and GI tissues. In brain, immunopositive cells were observed in hypothalamic areas known to be important in reproductive function and abundant axon terminals positive for pheonixin were seen in median eminence. Phoenixin exhibited high specific binding in pituitary and ovary, and injection of phoenixin into female rats led to the induction of early gene expression specifically in pituitary gonadotrophs. In dispersed primary rat pituitary cells, phoenixin potentiated the action of GnRH. Phoenixin treatment also led to the upregulation of LH Beta, FSH Beta, and GnRH receptor mRNAs in primary pituitary cells, as determined by RT-PCR. In order to determine the physiological relevance of phoenixin, siRNA directed against phoenixin was injected into the lateral cerebroventricle of female rats on diestrous days 1 and 2. Animals treated with siRNA had reduced levels of phoenixin peptide in the hypothalamus, as determined by RIA, and exhibited a delay in the occurrence of the subsequent estrus. Furthermore, treatment with phoenixin siRNA led to a decrease in GnRH receptor mRNA in the pituitary, suggesting that phoenixin [ldquo]sensitizes[rdquo] pituitary gonadotrophs by upregulating the transcription of the GnRH receptor. We therefore have discovered a novel reproductive hormone, phoenixin, that acts in the pituitary to possibly regulate expression of the GnRH receptor.[br][br]Nothing to Disclose: GLCY, R-ML, J-KC, CWB, NJD, SLD, AH, WKS 2012-06-25T11:15:00 310 2012-06-25T00:00:00 1899-12-30T11:15:00 1572 272 1899 OR25-1 OR200-03 Monday 1583 2012


1580 ENDO12L_OR25-2 ORAL SESSION: Female Reproduction: Gonadotropin Control [amp] Metabolic Responses (11:15 AM-12:45 PM) Depletion of Foxo Transcription Factors in Granulosa Cells Reveals Novel Mechanisms Controlling Follicle Growth and Death and Endocrine Regulation of Pituitary FSH Zhilin Liu, Diego H Castrillon, JoAnne S Richards Baylor College of Medicine, Houston, TX; University of Texas Southwestern Medical Center, Dallas, TX The Foxo transcription factors regulate multiple functions (metabolism, survival, differentiation and apoptosis) in a context- and developmental stage-specific manner by interacting with different signaling pathways. In the ovary FoxO1 and FoxO3 are highly expressed in granulosa cells of growing follicles and luteal cells, respectively, and are downstream targets of IGF1 and FSH. However, the development of ovarian follicles that is essential for the release of a fertilizable oocyte is regulated not only by the pituitary hormones FSH and LH and IGF1 but also by activins and BMPs. To determine the functional roles of Foxo1 and Foxo3 in this critical ovarian developmental process, we generated mice in which the [italic]Foxo1[/italic] and [italic]Foxo3 [/italic]genes were disrupted selectively in granulosa cells. Depletion of [italic]Foxo1/3[/italic] (but neither alone) in granulosa cells led to an infertile phenotype characterized by: 1) undetectable levels of serum FSH but not LH, 2) reduced expression of the pituitary [italic]Fshb[/italic] gene and transcriptional regulators of this gene and 3) ovarian production of an unknown factor(s) that suppresses pituitary cell [italic]Fshb[/italic] expression. Thus, loss of Foxo1/3 in granulosa cells dramatically alters a novel ovarian-pituitary endocrine feedback loop. Furthermore, independent of regulating pituitary FSH, depletion of [italic]Foxo1/3[/italic] altered the expression of specific genes in granulosa cells associated with follicle growth versus apoptosis by disrupting critical regulatory interactions of Foxo1/3 with the activin and BMP2 pathways, respectively. As a consequence, granulosa cell proliferation and apoptosis were decreased. These data provide the first evidence that Foxo1/3 divergently regulate follicle growth or death by interacting with the activin and BMP pathways in granulosa cells and by modulating pituitary FSH production.[br][br]Sources of Research Support: NIH-HD-16272, 16229 (JSR, ZL) and 48690 (DHC).[br][br]Nothing to Disclose: ZL, DHC, JSR 2012-06-25T11:30:00 310 2012-06-25T00:00:00 1899-12-30T11:30:00 309 272 1900 OR25-2 OR200-03 Monday 1584 2012


1581 ENDO12L_OR25-3 ORAL SESSION: Female Reproduction: Gonadotropin Control [amp] Metabolic Responses (11:15 AM-12:45 PM) Effects of Exogenous Human Gonadotropin Inhibitory Hormone (GnIH, RFRP3) on Luteinizing Hormone Secretion in Postmenopausal Women Jyothis Thomas George, Nicky Ferdyan, Karthik Srinivasan, Richard White, Richard A Anderson, Robert P Millar University of Edinburgh, Edinburgh, UK; Ferring Research Institute, San Diego, CA; University of Edinburgh, Edinburgh, UK Gonadotropin inhibitory hormone (GnIH) is proposed to be a putative counter-regulatory hormone to GnRH, inhibiting pituitary release of gonadotropins. It was described initially in birds but more recently GnIH peptide (RFRP3) has been demonstrated in the human hypothalamus [1]. Administration of GnIH decreases LH secretion in a number of species including ovariectomised sheep [2].[br]We hypothesized that administration of synthetic human RFRP-3 will decrease LH secretion in a comparable human model, ie post-menopausal women. Five women (mean age 58[plusmn]2 yrs, 12.4[plusmn]2.6 yrs since menopause, LH: 30.8[plusmn]2.9 IU/L, FSH: 78.7[plusmn]6.4 IU/L, estradiol: [lt]50 pmol/L) were infused with GMP-synthesized human GnIH at doses shown to inhibit LH pulsatility in ovariectomised sheep. Volunteers were admitted for 7 h of 10-min serum sampling to assess LH pulsatility. After 3 h of baseline sampling, an iv bolus of human GnIH (25% of hrly dose) was administered, followed by an human GnIH infusion for 3 h. [Delta]AUC of LH (the arithmetic difference of AUC over 3hr before and during the infusion) was calculated, and treatment effect analysed by paired t test.[br]Dose response studies were carried out in 2 women (1.5, 5, 15, 50 and 150 mcg/kg/hr or vehicle). Both two subjects showed a small decrease in LH secretion, with a maximal decrease at 15mcg/kg/hr and 50mcg/kg/hr (-9.7 and -10.7 IU/3hrs vs. 0.4 and 1.5 IU/L/3h during vehicle infusion). To assess circulating levels of human GnIH, plasma samples were collected in protease inhibitors and analysed by liquid chromatography tandem mass spectrometry. Plasma human GnIH concentrations of 3.6 nM (at 30 min, 50 mcg/kg/hr, n=2) and 11.3 nM (at 30 min, 150 mcg/kg/hr, n=2) were achieved. These concentrations have previously been shown to be similar to the affinity of the GnIH receptor, GPR147, and to inhibit LH in sheep.[br]The 50 mcg/kg/hr dose, selected for study in all volunteers based on data above, had no effect on LH pulse frequency but resulted in a small decrease in LH secretion, [Delta]AUC LH -9.9[plusmn]1.8 IU/L/3h vs. -3.7[plusmn]1.9 IU/L/3h during saline infusion (P = 0.020 n=5). No adverse events were observed.[br]In conclusion, a small but significant decrease in LH secretion was observed during brief human GnIH infusion in this first-in-man study. The effect was smaller than that observed in other species. Studies in eugonadal participants using longer duration of exposure are required to clarify inter-species differences and pharmacological potential of GnIH.[br][br][1] Ubuka T et al, PLoS One. 2009;4(12):e8400. [2] Clarke IJ, et. al, Endocrinology. 2008;149(11):5811-21.[br][br]Sources of Research Support: Medical Research Council (UK), Ferring Pharmaceuticals.[br][br]Disclosures: NF: Employee, Ferring Pharmaceuticals. KS: Employee, Ferring Pharmaceuticals. RW: Employee, Ferring Pharmaceuticals. Nothing to Disclose: JTG, RAA, RPM 2012-06-25T11:45:00 310 2012-06-25T00:00:00 1899-12-30T11:45:00 438 272 1901 OR25-3 OR200-03 Monday 1585 2012


1582 ENDO12L_OR25-4 ORAL SESSION: Female Reproduction: Gonadotropin Control [amp] Metabolic Responses (11:15 AM-12:45 PM) Relative Hypogonadism in Obese Women Is Explained by Blunted Pituitary Response to GnRH Lauren W Roth, Alex J Polotsky, Andrew P Bradford, Jennifer Lesh, Justin Chosich, Wendy M Kohrt, Nanette Santoro University of Colorado, Denver, Aurora, CO; University of Colorado, Denver, Aurora, CO The physiology behind the relative hypogonadotropic hypogonadism of female obesity (1) is unclear and may be due to hypothalamic or pituitary factors, or pharmacokinetic differences in gonadotropin processing.[br][bold]Methods:[/bold] A luteal phase frequent blood sampling study was undertaken in regularly menstruating obese and normal weight women (n=6 per group). The study included 12 hours of unstimulated monitoring (to evaluate endogenous hypothalamic-pituitary function), administration of GnRH 25 and 150ng/kg (to evaluate pituitary sensitivity), and overnight suppression with GnRH antagonist followed by recombinant LH (to evaluate LH pharmacokinetics). All subjects underwent DEXA. LH was measured with an immunofluorometric assay (DELFIA, Perkin-Elmer). LH pulsatility was evaluated using the Santen-Bardin method (2). Groups were compared using t tests or Mann-Whitney test as appropriate.[br][bold]Results:[/bold][br]All women were ovulatory. The obese group was significantly older than the normal weight group (33[plusmn]4 vs. 26[plusmn]4 years, p=0.01) and had a significantly higher BMI (33[plusmn]3 vs. 22[plusmn]1 kg/m[sup]2[/sup], p[lt]0.001). Unstimulated mean LH over 12 hours was significantly lower in the obese vs. normal weight women (4.1[plusmn]3.1 vs. 8.2[plusmn]6.4 IU, p[lt]0.001). LH pulse amplitude and frequency did not differ between groups. Higher percentage trunk fat on DEXA correlated inversely with lower mean LH level (r=-0.56, p=0.046).[br]After stimulation with GnRH, area under the curve for LH was significantly lower in the obese group (GnRH25ng/kg 1048[plusmn]36 vs. 1920[plusmn]46, p[lt]0.001, GnRH 150ng/kg 3343[plusmn]79 vs. 4853[plusmn]114, p=0.01). Time to peak LH after GnRH 25ng/kg was significantly reduced in the obese women (20 (20-20.8) vs. 30 (19.2-41.7) minutes, p=0.04). A similar finding was seen after GnRH 150ng/kg.[br]Peak LH, time to peak, half life, or decay constant for recombinant LH did not differ between groups after suppression with GnRH antagonist. Volume of distribution was slightly higher in the obese women (2.5[plusmn]0.7 vs. 3.3[plusmn]0.8, p=0.1).[br][bold]Conclusions:[/bold] The origin for the relative hypogonadotropic hypogonadism of obesity appears to result from decreased pituitary sensitivity to GnRH. The significantly lower unstimulated mean LH helps explain the corpus luteum insufficiency and subfertility in obese women. LH volume of distribution may play a minor role.[br][br](1) Jain A et al., J Clin Endocrinol Metab 2007;92:2468. (2)Santen RJ et al., J CLin Invest 1973;52:2617.[br][br]Sources of Research Support: NIH U54HD058155 Center for the Study of Reproductive Biology; Colorado Clinical and Translational Sciences Institute UL1RR025780; K24HD041978 to NS.[br][br]Nothing to Disclose: LWR, AJP, APB, JL, JC, WMK, NS 2012-06-25T12:00:00 310 2012-06-25T00:00:00 1899-12-30T12:00:00 536 272 1902 OR25-4 OR200-03 Monday 1586 2012


1583 ENDO12L_OR25-5 ORAL SESSION: Female Reproduction: Gonadotropin Control [amp] Metabolic Responses (11:15 AM-12:45 PM) Fat Mass and Obesity-Associated Gene Polymorphisms Do Not affect Metabolic Response to Hormone Therapy in Healthy Postmenopausal Women Ramon Bossardi Ramos, Gislaine Krolow Casanova, Poli Mara Spritzer Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil; Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil In the postmenopausal period, women experience an exponential increase in the risk of cardiovascular (CV) disease. The fat-mass and obesity-associated (FTO) gene variants are related with higher prevalence of CV risk and recently we shown an association of these FTO gene variants with the lipid accumulation product (LAP) index, which is also a marker of postmenopausal CV and metabolic risk (Ramos et al, Fertil Steril, 2011). The present study aimed to determine whether FTO gene single nucleotide polymorphisms (SNP) rs9939609 T [gt] A and rs8050136 A [gt] C or their haplotypes influence anthropometric and metabolic variables in recent postmenopausal women, before and after 6 months of hormone therapy (HT). In this randomized crossover study carried out in a university clinic, 86 postmenopausal women (51[plusmn]3 years, 22[plusmn]10 months since menopause) consulting for symptoms of estrogen deficiency were genotyped by real-time polymerase chain reaction for the two SNP of the FTO gene. Haplotypes were constructed from the combination of both polymorphisms, and their frequencies were inferred using the PHASE 2.1.1 program. Participants were clinically evaluated before and after 6 months of HT to determine body mass index and waist circumference, blood pressure, lipid profile, LAP, ultra sensitive C reactive protein (us-CRP), plasma glucose, and insulin. The genotypic distribution for rs993969 (TT: 44.2%, TA: 40.7%, AA: 10.5%) and rs8050136 (AA: 13.6%, AC: 35.8%, CC: 50.6%) was similar to previous studies. Five haplotypes were inferred in this sample (Ht1: CTCT, Ht2: CTCA, Ht3: CTAA, Ht4: ATAA, Ht5: AAAA). Haplotypes frequencies were 46.3% for Ht1, 42.6% for Ht2/Ht3/Ht4, and 10.9% for Ht5. The rs9939609 polymorphism was in almost complete linkage disequilibrium with the rs8050136 (|D[apos]| = 0[middot]953; r[sup2] = 0.876). In women with the homozygous polymorphic AA genotype of the single nucleotide polymorphisms rs9939609 and the wild AA genotype of the SNP rs8050136, LAP index and us-CRP were higher before HT in comparison with women with other genotypes from the same SNP group. There was no worsening of any of the anthropometric or metabolic variables, and LAP index improved slightly after HT in SNP rs9939609 (P = 0.03) and haplotype AAAA. No changes were observed after HT in SNP rs8050136. The presence of risk variants of the fat mass and obesity-associated gene seems not to exert a negative influence on the response to hormone therapy in apparently healthy early postmenopausal women.[br][br]Ramos RB et al., Fertil Steril 2011; 974-983.[br][br]Sources of Research Support: Brazilian Nacional Hormones and Women[apos]s Health.[br][br]Nothing to Disclose: RBR, GKC, PMS 2012-06-25T12:15:00 310 2012-06-25T00:00:00 1899-12-30T12:15:00 292 272 1903 OR25-5 OR200-03 Monday 1587 2012


1584 ENDO12L_OR25-6 ORAL SESSION: Female Reproduction: Gonadotropin Control [amp] Metabolic Responses (11:15 AM-12:45 PM) Mice Lacking Anti-M[uuml]llerian Hormone Are Protected Against Increased Adiposity and Glucose Intolerance with Increasing Age E Leonie AF van Houten, Piet Kramer, Anke Mcluskey, Bas Karels, Axel PN Themmen, Jenny A Visser Erasmus Medical Center, Rotterdam, Netherlands Polycystic ovary syndrome (PCOS), the most common endocrine disorder in women in their reproductive age, is defined by two out of the following three criteria: hyperandrogenism, oligo/anovulation, and polycystic ovaries. Polycystic ovaries contain an increased number of small antral follicles, which is suggested to result from an increased recruitment of primordial follicles. Enhanced recruitment of primordial follicles accompanied with an increased number of growing follicles is also observed in mice lacking anti-M[uuml]llerian hormone (AMH). Recently, we developed a mouse PCOS-model through chronic exposure to dihydrotestosterone (DHT). To determine the interaction between ovarian and metabolic function, we studied the impact of an increased growing follicle pool on the metabolic phenotype through DHT treatment of AMH knockout (AMHKO) mice.[br]Prepubertal AMHKO and wild type (wt) mice were treated with a 90-day continuous release DHT or placebo pellet. After treatment (90 days) mice were sacrificed (chronically-treated group, n=17) or allowed a 30-day wash-out period before sacrifice (persistent group, n=17). To determine an effect on metabolism, bodyweight and weight of adipose tissue was measured and an intraperitoneal Glucose Tolerance Test (IPGTT) (n=8-9 per group) was performed.[br]Bodyweight and weight of adipose depots increased to a similar extent in both chronically DHT-treated wt and AMHKO mice. In agreement, chronically DHT-treated AMHKO mice showed a comparable glucose intolerance as DHT-treated wt mice. In contrast, whereas both placebo- and DHT-treated wt mice of the persistent group showed a further increase in adiposity compared to the chronically-treated group, this did not occur in the AMHKO mice. In agreement, glucose intolerance aggravated in the placebo- and DHT-treated wt mice of the persistent group. In contrast, DHT withdrawal did not worsen glucose intolerance in DHT-treated AMHKO mice, although glucose tolerance remained impaired. In fact, both placebo- and DHT-treated AMHKO mice of the persistent group tended to have a better glucose tolerance than the chronically-treated AMHKO mice.[br]In conclusion, with increasing age, adiposity and glucose intolerance improved in placebo- and DHT-treated AMHKO mice, whereas treated wt mice display a more severe metabolic phenotype. These results suggest that an altered follicle dynamics in the ovary affect metabolism in time and may modulate the effect of androgen excess on metabolism.[br][br]Nothing to Disclose: ELAFvH, PK, AM, BK, APNT, JAV 2012-06-25T12:30:00 310 2012-06-25T00:00:00 1899-12-30T12:30:00 1545 272 1904 OR25-6 OR200-03 Monday 1588 2012


1585 ENDO12L_OR26-1 ORAL SESSION: New Insights into Thyroid Hormone Synthesis [amp] Action (11:15 AM-12:45 PM) Clinical Phenotype Associated with Mutation of Thyroid Hormone Receptor-[alpha]1 (TR[alpha]1) Alies van Mullem, Ramona van Heerebeek, Dionisios Chrysis, Edward Visser, Marco Medici, Maria Andrikoula, Agathocles Tsatsoulis, Robin Peeters, Theo J Visser Erasmus Medical Center, Rotterdam, Netherlands; University of Patras, Patras, Greece; University of Ioannina, Ioannina, Greece Thyroid hormone (TH) is essential for normal development and metabolism. The genomic action of TH is mediated by the binding of T3 to its nuclear receptors (TRs). Two genes, [italic]THRA[/italic] and [italic]THRB,[/italic] encode TRs, with TR[alpha]1 and TR[beta]1 as the major T3 binding isoforms. TR[alpha]1 is preferentially expressed in brain, bone and heart, whereas TR[beta]1 is considered the major isoform in liver, kidney, and thyroid. Heterozygous mutations in [italic]THRB [/italic]lead to resistance to TH (RTH). Patients with RTH have abnormally high serum TH and non-suppressed TSH levels, explained by the importance of TR[beta]2 in the negative feedback of TH at the pituitary and hypothalamus. Although TR[alpha]1 has already been characterized decades ago, no patients with mutations in this receptor have been identified until very recently. The phenotype of the first patients indicates that mutations in TR[alpha]1 are associated with abnormal TH but normal TSH levels, growth retardation, delayed bone development and constipation.[br]Here we describe the detailed clinical phenotype and treatment of two patients (father and daughter) with a heterozygous inactivating mutation in TR[alpha]1 (F397fs406X). Both patients have low (F)T4, high T3, low rT3 and normal TSH levels, in combination with high cholesterol, and low-normal GH and IGF-1 levels. Clinical characteristics include growth retardation due to a delayed bone development, mildly delayed mental and motor development, and constipation. Because of hypothyroid symptoms, patients were treated with LT4 resulting in normalization of (F)T4, persistently high T3 and suppression of TSH. LT4 also induced a transient increase in growth (in the girl), a normal defecation pattern and lowering of cholesterol. Despite the high T3 levels, the patients have a normal heart rate suggesting partial TH resistance of the heart. The highly elevated T3/rT3 ratio suggests altered peripheral TH metabolism. At 8 years of age, the girl was treated in addition with GH, which normalized IGF-1 but did not increase growth.[br]In conclusion, mutations in TR[alpha]1 are associated with abnormal TH levels, constipation and a delayed bone development. This phenotype is in agreement with the expression pattern of TR[alpha]1. Only partial normalization of the phenotype is achieved by LT4 treatment; growth and bone development remain delayed.[br][br]Nothing to Disclose: AvM, RvH, DC, EV, MM, MA, AT, RP, TJV 2012-06-25T11:15:00 360 2012-06-25T00:00:00 1899-12-30T11:15:00 2027 273 1905 OR26-1 OR38-01 Monday 1589 2012


1586 ENDO12L_OR26-2 ORAL SESSION: New Insights into Thyroid Hormone Synthesis [amp] Action (11:15 AM-12:45 PM) Thyroid Hormone Regulation of Sirtuin 1 (SIRT1) Expression and Implications for Integrated Responses to Energy Deprivation in Mice Aline Cordeiro, Luana Lopes Souza, Lorraine Soares Oliveira, Larissa Costa Faustino, Leticia Aragao Santiago, Flavia Fonseca Bloise, Tania Maria Ortiga-Carvalho, Norma Santos Almeida, Carmen Cabanelas Pazos-Moura Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil; Universidade Federal Rural do Rio de Janeiro, Seropedica, Brazil Sirtuin1 (SIRT1), a NAD[sup]+[/sup]-dependent deacetylase, has been importantly connected to beneficial effects elicited by calorie restriction, such as the extension of life span and reduction of oxidative stress. Physiological adaptation to starvation requires higher activity of SIRT1 and also the suppression of thyroid hormone (TH) production and action to achieve energy conservation. In the present study we investigated the relationship between these alterations during total or partial food restriction. 48h-fasting mice presented reduced serum TH and increased SIRT1 protein content (evaluated by western blotting) in liver and brown adipose tissue (BAT - p[lt]0.005), and physiological TH replacement to fasting mice prevented or attenuated the increment of SIRT1 in liver and BAT, respectively (p[lt]0.01),suggesting that TH suppression during fasting is necessary for the concomitant up-regulation of SIRT1. Therefore we investigated the effect of thyroid status on SIRT1 expression. Hypothyroid (PTU-containing diet) mice exhibited increased liver SIRT1 protein, while hyperthyroid (T3-treated) ones showed decreased SIRT1 in liver and brown adipose tissue (BAT, p[lt]0.0001). In the liver, decreased protein is accompanied by reduced SIRT1 activity (p[lt]0.05), measured employing a commercial kit, and no alteration in its mRNA, evaluated by real-time PCR. Hyperthyroid and hypothyroid mice exhibited increased and decreased food intake and body weight gain, respectively. Food-restricted hyperthyroid animals (pair fed to euthyroid group) exhibited hepatic SIRT1 protein levels intermediary between euthyroid and hyperthyroid mice fed [italic]ad libitum[/italic], but euthyroid mice food-restricted in the same percentage as hyperthyroid ones (40%) had no changes in liver SIRT1. Mice bearing a mutant thyroid hormone receptor beta, unable to bind T3, and thereby, resistant to thyroid hormone action, exhibited, as expected, high serum TH, but even so they presented increased hepatic SIRT1 protein and activity with no alteration in SIRT1 mRNA, further supporting a role for T3 acting via TR beta in the liver to decrease SIRT1 protein content. Altogether, these results suggest that TH decrease SIRT1 protein abundance, directly, and indirectly via food ingestion control, involving in the liver, the TR[beta] and probably, non-transcriptional mechanisms, with important implication to integrated metabolic responses to fasting, since the increase in SIRT1 protein requires the fasting-associated suppression of serum TH.[br][br]Sources of Research Support: FAPERJ, CNPq and CAPES.[br][br]Nothing to Disclose: AC, LLS, LSO, LCF, LAS, FFB, TMO-C, NSA, CCP-M 2012-06-25T11:30:00 360 2012-06-25T00:00:00 1899-12-30T11:30:00 929 273 1906 OR26-2 OR38-01 Monday 1590 2012


1587 ENDO12L_OR26-3 ORAL SESSION: New Insights into Thyroid Hormone Synthesis [amp] Action (11:15 AM-12:45 PM) Bioengineering of the Thyroid Lobe: Use of Its Stromal/Vascular Matrix as a Scaffold for Ex Situ Reconstruction Roberto Toni, Valentina Strusi, Nicoletta Zini, Davide Dallatana, Simone Mastrogiacomo, Annapaola Parrilli, Roberto Giardino, Giuseppe Lippi, Giulia Spaletta, Elena Bassoli, Andrea Gatto, Michele Iafisco, Monica Sandri, Anna Tampieri University of Parma School of Medicine, Parma, Italy; National Research Council (CNR) - Istituto Ortopedico Rizzoli, Bologna, Italy; Istituti Ortopedici Rizzoli, Bologna, Italy; Maggiore Hospital, Parma, Italy; University of Bologna, Bologna, Italy; University of Modena and Reggio Emilia, Modena, Italy; National Research Council (CNR), Faenza (RA), Italy Regenerative medicine of endocrine glands is one of the newest fields in biomedical research, and promises to become a primary strategy to treat a number of endocrine disorders. Using computational bioengineering we have recently suggested that the three-dimensional (3D) geometry of the thyroid, stromal/vascular matrix may act as an epigenetic guidance for growth and differentiation of developing thyrocytes (1-3). To test this hypothesis, we have bioengineered [italic]ex situ[/italic] (i.e. on the laboratory bench) a bioartificial rat thyroid lobe using its decellularized stromal/vascular matrix as a natural 3D scaffold, to be eventually recellularized with thyroid stem/precursor cells. Sprague-Dawley male rats (125-225 g) were used as thyroid donors, and lobe matrixes were prepared using a N[sub]2[/sub] freezing/Trypsin-EDTA/Triton-deoxycholate processing. Test matrixes were made electrondense, and analyzed by microTC (Skyscan 1172) to define their architecture. Primary thyroid cells were isolated after 72 h in primary monolayer culture, whereas presence of ABCG2-positive stem/precursor elements was determined using Western blotting. Following trypsinization, 250 - 450 x 10[sup]3[/sup] cells were harvested and dropped onto the empty follicular cavities of the inner matrix of single lobe halves for [ldquo]homing[rdquo]. Cultures were kept in static conditions up to 14 days. The recellularized matrixes were either fixed in aldheydes and analyzed with light, transmission, and scanning electron microscopy, or denaturated and processed for ABCG2 immunoblotting (polyclonal rabbit anti-human 1:2000, Cell Signalling). Culture supernatants were collected every 48 h for assessment of FT3 and FT4 by immuno-chemiluminescence (Beckman-Coulter). Complete decellularization and maintenance of the 3D native architecture of the lobe matrix were achieved. Thyroid-derived cells including thyrocytes, epithelial-mesenchymal elements, and stem/precursors were found to migrate inside matrix septa, self-assemble like follicule, and recellularize the decellularized native follicular spaces. Thyroid hormone secretion occurred for at least 7 days. These results show that the 3D matrix of the rat thyroid may guide both differentiated and stem-like elements to self-assemble into functional follicular units, up to the lobar recellularization. This raises the possibility that a bioartificial, immuno-tolerant thyroid gland be bioengineered [italic]ex situ[/italic] using autologous stem cells, and eventually transplanted.[br][br]Toni, R., Casa, C.D., Spaletta, G., Marchetti, G., Mazzoni, P., Bodria, M., Ravera, S., Dallatana, D., Castorina, S., Riccioli, V., Castorina, E.G., Antoci, S., Campanile, E., Raise, G., Rossi, R., Ugolotti, G., Martorella, A., Roti, E., Sgallari, F., Pinchera, A. The bioartificial thyroid: a biotechnological perspective in endocrine organ engineering for transplantation replacement. Acta Biomed. 78 (Suppl 1), 129-155, 2007. Toni, R., Casa, C.D., Bodria, M., Spaletta, G., Vella, R., Castorina, S., Gatto, A., Teti, G., Falconi, M., Rago, T., Vitti, P., Sgallari, F., 2008. A study on the relationship between intraglandular arterial distribution and thyroid lobe shape: implications for biotechnology of a bioartificial thyroid. Ann. Anat. 190, 432-441, 2008. Toni R, Tampieri A, Zini N, Strusi V, Sandri M, Dallatana, Spaletta G, Bassoli E, Gatto A, Ferrari A, Martin I. Ex situ bioengineering of bioartificial endocrine glands: a new frontier in regenerative medicine of soft tissue organs. Annals of Anatomy 193,381-394,2011.[br][br]Sources of Research Support: Grants PRIN082008ZCCJX4,FIRB2010RBAP10MLK7.[br][br]Nothing to Disclose: RT, VS, NZ, DD, SM, AP, RG, GL, GS, EB, AG, MI, MS, AT 2012-06-25T11:45:00 360 2012-06-25T00:00:00 1899-12-30T11:45:00 332 273 1907 OR26-3 OR38-01 Monday 1591 2012


1588 ENDO12L_OR26-4 ORAL SESSION: New Insights into Thyroid Hormone Synthesis [amp] Action (11:15 AM-12:45 PM) Thyroid Hormone Stimulates Hepatic Lipophagy Coupled with Fatty Acid Metabolism Rohit Anthony Sinha, Seo-Hee You, Christopher B Newgard, Mitchell A Lazar, Paul M Yen Duke-NUS Graduate Medical School, Singapore, Singapore; Perelman School of Medicine at the University of Pennsylvania School of Medicine, Philadelphia, PA; Duke University Medical Center, Durham, NC Thyroid hormones have been known to exert powerful catabolic effects leading to weight loss for many years. While T3 stimulation of beta-oxidation of fatty acids in the liver has been well-described, little is known about the mechanisms by which this occurs. Recently, we found that T3 surprisingly stimulates autophagy in human hepatic cells in vitro and in mouse livers in vivo. In particular, T3 increased LC3IIB and decreased p62 expression levels in HepG2TR[alpha] cells and hepatocytes. Moreover, chloroquine treatment demonstrated that T3 increased overall autophagic flux. Electron microscopy and Bodipy-staining studies of hepatic cells in culture and in vivo showed that T3-induced autophagy remarkably involved the formation of lipophagosomes that ingest lipids from fat droplets and then fuse with lysosomes before delivery to mitochondria. T3-mediated lipophagy also was associated with increased beta-oxidation as T3 increased Cpt-1[alpha] expression. Metabolomic analysis of hepatic acylcarnitines demonstrated increased flux of beta-oxidation metabolites in hyperthyroid mice. Furthermore, knockdown studies using ATG5 siRNA showed that T3-dependent beta-oxidation was abrogated when autophagy was blocked. We also observed that T3 stimulated intracellular pAMPK and SIRT1 levels, both of which are known to promote autophagy. We next found that knock-in mice that over-express a dominant negative co-repressor, NCoR containing a deacetylase activating domain mutation (DADm), exhibited altered T3-mediated hepatic autophagy and metabolomic lipid profile. Finally, T3-mediated autophagy reduced fatty acid-induced lipotoxicity in HepG2 cells. Taken together, our findings show that T3 likely plays a critical role in the regulation of fatty acid metabolism by coupling autophagy with beta-oxidation. This action depends upon normal corepressor function. They also suggest that T3 or its analogs may be useful for the treatment of NAFLD, a common condition with high morbidity caused by excess intra-hepatic fat accumulation.[br][br]Sources of Research Support: This work was supported by funded by NIH DK 43806 (to MAL), the Nuclear Receptor Signaling Atlas NIH U19DK/HL/ES 62434 (to CBN and MAL), as well as an ADA mentored research fellowship (to SHY and MAL) This work also was supported by Duke-NUS Graduate Medical School Faculty Funds (PMY) sponsored by the Ministry of Health, Ministry of Education, and Ministry of Trade, Singapore and A*StaR.[br][br]Nothing to Disclose: RAS, S-HY, CBN, MAL, PMY 2012-06-25T12:00:00 360 2012-06-25T00:00:00 1899-12-30T12:00:00 401 273 1908 OR26-4 OR38-01 Monday 1592 2012


1589 ENDO12L_OR26-5 ORAL SESSION: New Insights into Thyroid Hormone Synthesis [amp] Action (11:15 AM-12:45 PM) Monocarboxylate-Transporter-8 and Type 3 Deiodinase in Axon Terminals of the Median Eminence Allow System-Specific Regulation of T3-Action in Hypothalamic Hypophysiotropic Neurosecretory Neurons Petra Mohacsik, Imre Kallo, Barbara Vida, Aniko Zeold, Zsuzsanna Bardoczy, Erzsebet Farkas, Andrea Kadar, Ann M Zavacki, Rafael Arrojo e Drigo, Liping Dong, Ronald M Lechan, Antonio C Bianco, Zsolt Liposits, Csaba Fekete, Balazs Gereben Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary; Brigham and Women[apos]s Hospital and Harvard Medical School, Boston, MA; University of Miami Miller School of Medicine, Miami, FL; Tupper Research Institute and Tufts Medical Center, Boston, MA The hypothalamic neurosecretory neurons are crucial regulators of metabolism, growth, stress and reproduction. These cells represent a major target of hypothalamic thyroid hormone signaling. Monocarboxylate-transporter-8 (MCT8)-mediated transport of thyroid hormone across the plasma membrane followed by its inactivation by type 3 deiodinase (D3) represents two limiting factors regulating neuronal T3 levels. We previously found D3-immunoreactivity in axon varicosities of the external zone of the rat hypothalamic median eminence where the enzyme was localized to dense-core vesicles in hypophysiotropic axon varicosities. We investigated whether MCT8 could allow T3 influx directly to axon terminals and quantified the co-existence of MCT8 and D3 in specific neurosecretory systems using immunofluorescent, confocal microscopy. MCT8-immunoreactivity was observed on the surface of the vast majority of all types of hypophysiotropic terminals in the median eminence. However, D3 was present in 71.8[plusmn]3.8% of axon varicosities containing GnRH, 63.2[plusmn]7.5% of CRH and 64.2[plusmn]2.7% of GHRH, but only in 26.6[plusmn]5.0% of that containing TRH, while completely absent from somatostatin-containing neurons. To determine whether D3 is functionally active in this location, we studied whether D3-mediated catalysis and T3-mediated D3 regulation is functional in the axonal compartment. D3 homodimerization, a pre-requisite for D3 activity, was detected in cellular processes of GnRH expressing GT1-7 cells using a Bimolecular Fluorescence Complementation approach. Furthermore, D3 activity was induced 4-fold in the median eminence of hyperthyroid rats, showing that axonal D3 responds to an elevated T3 influx. These observations suggest fine-tuned and system-specific thyroid hormone metabolism in different types of hypophysiotropic axons that involve MCT8-mediated transport and D3-catalyzed inactivation. This pathway may represent a novel, regulatory pathway for T3-mediated hypothalamic function with potential effects on the control metabolism, growth, stress and reproduction.[br][br]Sources of Research Support: This work was supported by the National Science Foundation of Hungary (OTKA K81226 and K81845), EU FW7 Health-F2-2010-259772, Lend[uuml]let Award of the Hungarian Academy of Sciences and NIH DK-37021.[br][br]Nothing to Disclose: PM, IK, BV, AZ, ZB, EF, AK, AMZ, RAeD, LD, RML, ACB, ZL, CF, BG 2012-06-25T12:15:00 360 2012-06-25T00:00:00 1899-12-30T12:15:00 608 273 1909 OR26-5 OR38-01 Monday 1593 2012


1590 ENDO12L_OR26-6 ORAL SESSION: New Insights into Thyroid Hormone Synthesis [amp] Action (11:15 AM-12:45 PM) Lacking TSH Receptor by Small Interfering RNA or Knockdown Decreases the Expression of HMGCoA Reductase in the Mouse Liver Xiujuan Zhang, Baoxiang Hu, Yongfeng Song, Tingting Wang, Ling Gao, Jiajun Zhao Provincial Hospital Affiliated to Shandong University, Jinan, China; Shandong Academy of Clinical Medicine, Jinan, China; Provincial Hospital Affiliated to Shandong University, Jinan, China [bold]Objective[/bold]:[br]The elevated total cholesterol (TC) level is often imputed to thyroid hormone (TH) deficiency in Hypothyroidism. However, subclinical hypothyroidism, TH stays within its normal range, has still elevated serum TC and TSH levels. It raise a question of whether TSH associated with cholesterol. Here our aim was to observe the effect of TSH on hepatic HMGCR, rate-limiting enzyme of cholesterol synthesis, in mice[italic] in vivo[/italic].[br][bold]methods: [/bold][italic]Model 1.[/italic] Male Babc/l mice in SCH group were given 8 mg/kg[middot]d methimazole in the drinking water, mice in the control group were given an equal volume of distilled water. At the end of 5 months, all mice were sacrificed.[br][italic]Model 2.[/italic] Male Babc/l mice in the TSHR-siRNA group were received a high-pressure injection of siRNA by tail-vein injection repeated 7 times in a 15-days, mice in the control were given an equal volume of distilled water.[br][italic]Model 3.[/italic] B6.129S1-Thrb[sup]tm1Df[/sup]/J mice genetically deficient for the TSHR were maintained on thyroid-supplemented chow from birth. Male mice (aged 6[ndash]8 week) were sacrificed.[br]Protein and mRNA expression was tested using Western blotting and real-time PCR. Tests for HMGCR activity, cholesterol content and TH were finished using HLPC-MS, specific kit and RIA method, respectively.[br][bold]Results: [/bold]Model 1. Compared to the control, SCH mice showed 154.2 % increases in the circulating levels of TSH([italic]p[/italic][lt]0.05), but no changes of FT3 ([italic]p[/italic][gt]0.05). The TC levels were increased 17.5 % and 19.1% ([italic]both p[/italic][lt]0.05) in the serum and liver, respectively. The HMGCR protein and its activity were enhanced by 49 % and 65.6 % (both [italic]p[/italic] [lt]0.05), respectively.[br]Model 2. TSHR siRNA-applied mice, the TSHR expression of mRNA and protein was significantly inhibited in liver ([italic]p[/italic][lt]0.05) not in thyroid. The HMGCR protein level in liver was decreased 63% ([italic]p[/italic][lt]0.05), but LDLR protein showed no obvious change ([italic]p[/italic][gt]0.05). The TC levels were decreased 35.6% and 45 % (both p[lt]0.05) in the serum and liver, respectively.[br]Model 3 Compared with the wild type mice, the serum TH levels were no change ([italic]p[/italic] [gt]0.05), while the serum TC level was declined 35% ([italic]p [/italic][lt]0.05) and the liver HMGCR expression was decreased 42.1%([italic]p[/italic][lt]0.05), respectively, in the TSHR-knockout mice.[br][bold]Conclusion[/bold]: Inhibition of THSR expression by siRNA or knock-down significantly attenuates HMGCR expression and HMGCR-mediate cholesterol level in the mice liver, further indicating the function of TSH on liver cholesterol synthesis.[br][br]Sources of Research Support: This research is supported by National Natural Science Foundation of China(81170794, 30971409), Natural Science Foundation (ZR2009CZ009) and the international cooperation grant (2011) of Shandong Province of china.[br][br]Nothing to Disclose: XZ, BH, YS, TW, LG, JZ 2012-06-25T12:30:00 360 2012-06-25T00:00:00 1899-12-30T12:30:00 1198 273 1910 OR26-6 OR38-01 Monday 1594 2012


1591 ENDO12L_OR27-1 ORAL SESSION: Parathyroid Hormone Disorders (11:15 AM-12:45 PM) Cellular Heterogeneity in the Function and Clonal Origin of Parathyroid Adenomas Implies Distinct Disease Etiologies in Primary Hyperparathyroidism Yuhong Shi, Joyce Hogue, Darshana Dixit, James Koh, John A Olson, Jr Duke University Medical Center, Durham, NC; Duke University Medical Center, Durham, NC; University of Maryland School of Medicine, Baltimore, MD Sporadic parathyroid adenomas (PA) are thought to arise from clonal expansion of a single transformed parathyroid cell progenitor. We report that human PA are comprised of heterogeneous cellular populations with distinct functional properties and variable clonal origin. We find that PA contain functionally discrete and separable subpopulations (designated P4 and P5) that can be distinguished using a quantitative live-cell fluorescence-based readout of calcium responsiveness. Electron microscopy ultrastructure analysis reveals that P4 cells are parathyroid chief cells whereas P5 cells are parathyroid oxyphil cells. P5 cells produce approximately 50% more PTH than P4 cells on a per cell basis under normocalcemic conditions. CASR transcript abundance, protein subcellular distribution and the fraction of cells expressing immunofluorescence-visualized CASR protein is equivalent in the P4 and P5 populations, but P5 cells display significantly greater calcium responsiveness ([gt]2-fold) and account for the majority of overall calcium flux response in cultured parathyroid cells. We examined PA clonality using both intact tumor samples and preparatively sorted P4 and P5 cells, with patient-matched peripheral blood lymphocyte DNA as an allelic control. Of fifteen samples assayed, we found 7 to be polyclonal and 8 to be monoclonal. To confirm these results we employed laser-captured microdissection to isolate parathyroid adenoma cells from an additional five samples. Three of the 5 specimens were polyclonal, while 2 of 5 exhibited monoclonality. In the monoclonal cases, both the P4 and P5 cells contained the same maternally imprinted allele of the X-linked human androgen receptor (HUMARA) gene, indicating that these populations arose from a common progenitor. Conversely, in the polyclonal specimens both the P4 and P5 populations were found to contain equal representation of each imprinted HUMARA allele, indicating the non-clonal provenance of both cell types. These data demonstrate the presence of functionally distinct subclasses of parathyroid cells within parathyroid primary adenomas and provide evidence that adenomas associated with HPT can arise by both clonal and polyclonal mechanisms. The differential provenance, biochemical activity, and relative abundance of these parathyroid adenoma subpopulations may reflect distinct mechanisms of disease etiology that could contribute to heterogeneity in clinical presentation, therapeutic response, and outcome in PHPT.[br][br]Sources of Research Support: NIH GRANT 1R01DK088188-01 (JAO and JK).[br][br]Nothing to Disclose: YS, JH, DD, JK, JAO 2012-06-25T11:15:00 332 2012-06-25T00:00:00 1899-12-30T11:15:00 1925 274 1911 OR27-1 OR06-01 Monday 1595 2012


1592 ENDO12L_OR27-2 ORAL SESSION: Parathyroid Hormone Disorders (11:15 AM-12:45 PM) Long-Term Follow-Up of Patients with Hypoparathyroidism [mdash] A Cohort Study Deborah M Mitchell, Susan Regan, Michael R Cooley, Kelly B Lauter, Michael C Vrla, Carolyn B Becker, Sherri-Ann M Burnett-Bowie, Michael Mannstadt Massachusetts General Hospital, Boston, MA; Massachusetts General Hospital, Boston, MA; Massachusetts General Hospital, Boston, MA; Brigham and Women[apos]s Hospital, Boston, MA Hypoparathyroidism is a rare disorder with complications including symptomatic hypo- and hypercalcemia and renal calcifications. Large cohort studies to better describe this population are lacking. Using an electronic clinical data registry, we identified 120 patients with permanent hypoparathyroidism treated within the Partners Healthcare System and performed detailed chart reviews. [underline]Cohort Description[/underline]: The cohort was 73% female with an age of 53[plusmn]18 years (range 2-87) and disease duration of 17[plusmn]16 years (range 1-58). The cause of hypoparathyroidism was post-surgical (66%), autoimmune (8%), calcium-sensing receptor mutations (4%), DiGeorge syndrome (3%), Kearns-Sayre syndrome (1%), and idiopathic (18%). [underline]Treatment[/underline]: Average calcium supplementation was 2.0[plusmn]1.5g/day. 88% of patients were prescribed calcitriol (0.53[plusmn]0.48 mcg/day) and 6% were prescribed high-dose vitamin D. 20% of patients were also prescribed a thiazide. [underline]Laboratory and Imaging Data[/underline]: Time-weighted mean serum calcium (Ca) and phosphorus levels were 8.4[plusmn]0.7 and 4.2[plusmn]0.8 mg/dL respectively. Linear interpolation was used to determine the proportion of time spent with Ca in the therapeutic range (7.5-9.5 mg/dL) with a median of 87% (range 13-100). Post-surgical patients had higher average Ca levels (8.6 vs. 8.1 mg/dL, p[lt]0.001) and greater proportions of time in the therapeutic range (median 93 vs. 82%, p=0.013) compared to patients with non-surgical etiologies. 53 patients (44%) had a 24 hour urine Ca collection, of whom 20 (38%) had a level [ge] 300 mg. eGFRs were calculated and were substantially lower than age-related population norms (p[lt]0.01). In multivariate analysis, predictors of lower eGFR included age, disease duration, and proportion of time with relative hypercalcemia (Ca [gt] 9.5 mg/dL). 54 patients had renal imaging, of whom 31% had renal calcifications, and 31 had head imaging, of whom 52% had basal ganglia calcifications. [underline]Complications[/underline]: 33% of patients were evaluated in an emergency department or hospitalized for complications of hypoparathyroidism, including 7% in the last year of observation. Renal complications included eGFR [lt]60 mL/min/1.73 m[sup]2[/sup] (41%), nephrolithiasis or nephrocalcinosis (16%), and renal transplantation (2%). [underline]Conclusions[/underline]: This study reports the largest cohort of patients with hypoparathyroidism to date. Our data show that hypoparathyroidism and its treatment carry an unexpectedly high burden of disease.[br][br]Sources of Research Support: NIH grant T32DK007028-37 awarded to DMM; NIH grant K08-DK081669 awarded to MM.[br][br]Nothing to Disclose: DMM, SR, MRC, KBL, MCV, CBB, S-AMB-B, MM 2012-06-25T11:30:00 332 2012-06-25T00:00:00 1899-12-30T11:30:00 1099 274 1912 OR27-2 OR06-01 Monday 1596 2012


1593 ENDO12L_OR27-3 ORAL SESSION: Parathyroid Hormone Disorders (11:15 AM-12:45 PM) Age and Gender Affect the Development of PTH Resistance in a Novel Model of Chronic Hyperparathyroidism Zhiqiang Cheng, Nathan Liang, Tsui-Hua Chen, Alfred Li, Christian Santa Maria, Michael You, Hanson Ho, Fuqing Song, Daniel Bikle, Chialing Tu, Dolores Shoback, Wenhan Chang University of California, San Francisco, CA Age and gender are critical determinants of manifestations and complications of human primary hyperparathyroidism (HPTH). To study age and gender effects on responses of target organs (kidney, bone, intestine) to chronic PTH excess in vivo, we generated mice with heterozygous Casr gene knockout (Het-KO) in parathyroid (PT) cells. Het-KO mice showed mild early-onset HPTH with modest hypercalcemia at 2 months of age. Serum PTH levels rose with age in Het-KO mice with a strong gender influence. At 12 months of age, PTH rose by 100 and 200% in male and female Het-KO mice, respectively, vs control mice. Interestingly, a concurrent reduction (by 10-15%) in serum Ca[sup]2+[/sup] (sCa) occurred, indicating a waning hypercalcemic effect of PTH in aging mice. Consistent with reduced calcemic effects of PTH, pPCR analyses of RNA from kidney and duodenum epithelia showed reduced ability with age of chronic HPTH to enhance renal expression of CYP27b1 and TRPV5 and intestinal expression of 1,25-dihydroxyvitamin D (1,25D)-responsive Ca[sup]2+[/sup]-transport proteins (TRPV6, calbindin, and PMCA) and the vitamin D receptor in 12- vs 3-month-old mice. This may be mediated by the observed decreased renal expression of the PTH1 receptor by qPCR and the subsequent decrease in 1,25D levels. In response to the lowering of sCa with age, PT glands of Het-KO mice showed an increase in their Ca[sup]2+[/sup]-sensing ability, indicated by a left-shifted Ca[sup]2+[/sup]-PTH secretion set-point in 12- vs 3-month-old glands in a PT organ culture model. This compensation was significantly less in female vs male Het-KO mice [ndash] such that more aberrant increases in sPTH were seen in 12-month-old female Het-KO mice. Skeletal responses to HPTH were age-, gender-, and site-dependent. In male Het-KO mice, HPTH enhanced trabecular bone mass at [ge] 3 months of age by microCT which was mainly due to decreased bone turnover and osteoclast numbers on resorption surfaces by histomorphometry. HPTH produced catabolic effects on cortical bone at all ages tested due to delayed bone mineralization. In contrast in females, HPTH was catabolic in trabecular bone, due to increased bone turnover and osteoclast numbers, and slightly anabolic in cortical bone, due to enhanced mineralization at 3 months. These effects were reversed at 12 months of age. We conclude that gender- and age-dependent changes in responses to calciotropic hormones by target organs contribute to the manifestations of HPTH and may underlie the higher disease prevalence in women.[br][br]Sources of Research Support: NIH Grants AG21353, AR50662 awarded to WC, AR050023 awarded to DB, AR055588 awarded to DS, and AR056256 awarded to CLT; Department of Veteran Affairs Merit Review awarded to DB and DS, and Research Education Advancement Program in Bone Disease awarded to WC, DB, and DS.[br][br]Nothing to Disclose: ZC, NL, T-HC, AL, CSM, MY, HH, FS, DB, CT, DS, WC 2012-06-25T11:45:00 332 2012-06-25T00:00:00 1899-12-30T11:45:00 905 274 1913 OR27-3 OR06-01 Monday 1597 2012


1594 ENDO12L_OR27-4 ORAL SESSION: Parathyroid Hormone Disorders (11:15 AM-12:45 PM) Functional Analysis and the Effect of Allosteric Modulator (NPS R568 and NPS 2143) to Loss-of-Function and Gain-of-Function Mutations of Calcium-Sensing Receptor Akie Nakamura, Katsura Ishizu, Toshihiro Tajima Hokkaido University, Sapporo, Japan Background:[br]The calcium-sensing receptor (CASR) belongs to a family C group of G protein-coupled receptor. CASR expresses in several organs which relate to calcium homeostasis. CASR in parathyroid glands regulate PTH secretion response to extracellular calcium levels. Activating mutations cause autosomal dominant hypoparathyroidism (ADH), and inactivating mutations cause neonatal severe hyperparathyroidism (NSHPT) and familial hypocalciuiric hypercalcemia (FHH). In these conditions, it is sometimes difficult to control their calcium levels within appropriate range. Recently, allosteric modulator is focused as new drug treatment for the patients with CASR mutations. Positive allosteric modulators of the CASR are called calcimimetics and negative allosteric modulators of the CASR are called calcilytics. In this study, the effect of NPS R-568 (calcimimetics) and NPS 2143 (calcilytics) to the CASR mutants, which were identified in ADH and FHH patients, was studied in vitro.[br]Methods:[br]Mutations of CASR from three ADH and two FHH Japanese patients were investigated. Functional consequences on Gi-MAPK and Gq-IP3 pathways and cell surface expression were determined. Furthermore, we evaluated whether or not NPS R-568 and NPS 2143 rescue the activity of mutant CASRs.[br]Results:[br]We identified three gain-of-function mutations (S122C, P569H and I839T) in ADH patients and two loss-of-function mutations (A110T and R172G) in FHH patients. S122C, P569H and I839T showed leftward shift in its dose-response curves response to increasing extracellular calcium concentrations in both Gi-MAPK and Gq-IP3 pathways in. By contrast, A110T and R17G showed rightward shift in its dose-response curve in both Gi-MAPK and Gq-IP3 pathways.[br]NPS R-568 rescued the capacity of signal transduction of A110T and R172G without increase of cell surface expression. On the other hand, NPS 2143 decreased the enhanced activity of S122C, P569H, I839T and A843E which cause a constitutive activation, but the response of A843E was milder than other mutants.[br]Conclusions:[br]Our results indicated that calcimimetics and calcilytics should be a new treatment for severe FHH and ADH patients. However, as shown in our experiment, there is possibility that the degree of functional recovery is different among mutants. Therefore, in vitro analysis of each mutant CASR by allosteric modulators is necessary.[br][br]Nothing to Disclose: AN, KI, TT 2012-06-25T12:00:00 332 2012-06-25T00:00:00 1899-12-30T12:00:00 150 274 1914 OR27-4 OR06-01 Monday 1598 2012


1595 ENDO12L_OR27-5 ORAL SESSION: Parathyroid Hormone Disorders (11:15 AM-12:45 PM) Radiofrequency Ablation of Pulmonary Metastases in Parathyroid Carcinoma: An Alternative Therapy for Severe Refractory Hypercalcemia Delmar E Lourenco, Jr, Ana O Hoff, Ana Elisa E Alcantara, Regina M Martins, Pedro Henrique S Correa, Marcos Menezes University of S[atilde]o Paulo School of Medicine, S[atilde]o Paulo, Brazil; Cancer Institute of S[atilde]o Paulo State (ICESP), S[atilde]o Paulo, Brazil Parathyroid carcinoma (PCA) is a rare disease accounting for 0.005% of all malignances and less than 2% of primary hyperparathyroidism (HPT) cases. HRPT2 gene mutations play a central role in the pathogenesis of parathyroid carcinoma and are observed not only in sporadic but also in familial cases. Parathyroid carcinoma is an aggressive disease associated with significant morbidity and mortality. Patients present with severe hypercalcemia, renal insufficiency and debilitating bone manifestations including fractures and brown tumors. Surgery is the main treatment but unfortunately recurrent disease is extremely common. With the goal of disease control and improvement of hypercalcemia, the treatment of metastatic PCA includes surgery of metastatic lesions, bisphosphonates, radiotherapy, chemotherapy and calcimimetics. Despite a multidisciplinary approach, death from uncontrolled hypercalcemia is common. Recently, radiofrequency ablation of metastatic PCA has been proposed as an alternative approach to control disease. In this report, we describe 3 patients with metastatic parathyroid carcinoma that failed conventional therapy and underwent radiofrequency ablation of pulmonary lesions. All patients had severe hypercalcemia resistant to hydration, calcitonin and zoledronic acid. One patient was treated with cinacalcet but failed to tolerate it. All patients had multiple bilateral pulmonary metastases not amenable for surgery and severe bone disease (fractures, brown tumors).[br]Pre-procedure serum calcium ranged from 14.8 mg/dL to 19.3 mg/dL and serum PTH ranged from 1519 pg/mL to 6118 pg/mL. Radiofrequency ablation was performed in at least two steps to treat lesions from both lungs. Normalization of serum calcium and significant reduction of PTH was observed in all 3 patients. Information on long-term control is available in 1 patient who remained stable for 7 months when a repeat ablation was performed and resulted in calcium normalization and disease control for another 8 months. The other 2 patients were recently treated. Conclusion: Radiofrequency ablation of metastatic parathyroid carcinoma to lungs is an alternative approach to control refractory hypercalcemia.[br][br]Sources of Research Support: Sources of Research Support: This study was supported by Sao Paulo State Research Foundation (FAPESP) Grants #2008/58552.[br][br]Nothing to Disclose: DEL, AOH, AEEA, RMM, PHSC, MM 2012-06-25T12:15:00 332 2012-06-25T00:00:00 1899-12-30T12:15:00 1662 274 1915 OR27-5 OR06-01 Monday 1599 2012


1596 ENDO12L_OR27-6 ORAL SESSION: Parathyroid Hormone Disorders (11:15 AM-12:45 PM) The Impact of Obesity on the Presentation of Primary Hyperparathyroidism Hien T Tran, Naim Maalouf University of Texas Southwestern Medical Center, Dallas, TX [underline]Background[/underline]: Obesity has been associated with vitamin D deficiency, elevated serum parathyroid hormone (sPTH), and higher prevalence of primary hyperparathyroidism (PHPT). The impact of vitamin D deficiency and elevated sPTH on the presentation of PHPT in obese patient is not entirely understood.[br][underline]Objective[/underline]: To compare obese (OB) and non-obese (NO) PHPT patients and assess the role of vitamin D deficiency in the relationship between obesity and PHPT.[br][underline]Methods[/underline]: Records from consecutive patients with PHPT who underwent parathyroidectomy between 2003-2009 by a single surgical group were reviewed. Pre-operative data on demographic, historical, laboratory, and radiological findings were reviewed and compared in OB and NO patients.[br][underline]Results[/underline]: Records of 231 patients with PHPT were included. Forty-seven percent were obese and 79% were women. Age, gender and race distribution between the two groups were similar. Mean BMI was 25.1[plusmn]3.3 and 35.6[plusmn]5.3kg/m[sup]2[/sup] in the NO and OB groups, respectively. Serum calcium and 25-hydroxyvitamin D were similar (Ca: 10.9[plusmn]0.7mg/dL in NO vs. 11.0[plusmn]0.7mg/dL in OB, p=0.4; 25(OH)D: 25[plusmn]16ng/mL in NO vs. 23[plusmn]11ng/mL in OB, p=0.43). sPTH correlated positively with BMI. Compared with the NO group, OB subjects had significantly higher sPTH (159[plusmn]75 vs. 133[plusmn]72pg/mL, p=0.01). Prevalence of kidney stones was significantly higher in the OB group (36.1% vs. 19.5% in NO, p=0.02) and hypercalciuria (urine calcium over 400mg/24hr) was more common in the OB group (39% vs. 24% in NO group, p=0.04). Despite higher sPTH and no difference in 25(OH)D levels, OB patients showed a higher bone mineral density and lower rates of osteoporosis (24.6% (OB) vs 43.3% (NO), p=0.03). Differences between OB and NO in sPTH, prevalence of nephrolithiasis and osteoporosis persisted after adjustment for age, race, eGFR and gender. Adjustment for serum 25-OH-D did not alter the relationship between obesity and nephrolithiasis or osteoporosis.[br][underline]Conclusions[/underline]: OB patients with PHPT are more likely to present with hypercalciuria and nephrolithiasis and less likely to present with osteoporosis when compared with NO PHPT patients with similar serum calcium. These differences are independent of age, gender, eGFR and race, and cannot be entirely explained by vitamin D deficiency and resultant 2ry hyperparathyroidism.[br][br]Nothing to Disclose: HTT, NM 2012-06-25T12:30:00 332 2012-06-25T00:00:00 1899-12-30T12:30:00 1418 274 1916 OR27-6 OR06-01 Monday 1600 2012


1597 ENDO12L_OR28-1 ORAL SESSION: Male Hypogonadism: Predisposition [amp] Risks (11:15 AM-12:45 PM) Association between Serum Testosterone Levels and Vascular Calcification in Community-Dwelling Men: The Framingham Heart Study Thomas G Travison, Shehzad Basaria, Shalender Bhasin, Anh-Hoa Nguyen, Joseph Massaro, Warren J Manning, Ramachandran S Vasan, Christopher J O[apos]Donnell Boston University School of Medicine, Boston, MA; Boston University School of Public Health, Boston, MA; Harvard Medical School, Boston, MA; National Heart, Lung and Blood Institute, Framingham, MA Background: Observational associations between male circulating testosterone (T) levels and cardiovascular disease and mortality have been widely reported, but the relationship between T and atherosclerosis is poorly understood. Here we report an analysis of association between serum testosterone and vascular calcification in adult men.[br]Methods: Data were available on 1678 participants belonging to Generation 2 (examination 7) and Generation 3 (examination 1) of the Framingham Heart Study cohort, and were obtained in clinical study visits conducted between 1998 and 2008. Early-morning serum total testosterone (TT) was measured by liquid chromatography-tandem mass spectrometry, and free testosterone was calculated (cFT) using mass action equations. Sex hormone-binding globulin (SHBG) was measured using immunofluorometric assay. Thoracic aortic calcification (TAC), abdominal aortic calcification (AAC), and coronary artery calcification (CAC) were measured by computed tomography scan. Covariables considered included age, body mass index (BMI), smoking, fasting lipids, and comorbid conditions. Estimates of association were obtained by Tobit regression, which acknowledges the influence of floor effects on outcomes.[br]Results: Subjects had mean (SD) age 49 (10) y with range 31-80 y; mean (SD) BMI was 28 (5) kg/m2. Mean (SD) TT, cFT, and SHBG were 616 (224) ng/dl, 111 (45) pg/ml, and 46 (23) nmol/l, respectively. TT and cFT exhibited highly significant positive, and SHBG negative, association with vascular calcification at all sites (p[lt]0.001). After adjusting for age, SHBG displayed no association with vascular calcification. Scaled to the mean sample CAC, a 100-ng/dl between-subject increase in TT was associated with a mean (SE) age-adjusted decrease in CAC of -8.6% (4.5%), p=0.04. Similarly, a 10 pg/ml positive difference in cFT was associated with a mean (SE) CAC decrease of -5.1% (2.5%), p=0.04. Trends of similar magnitude were observed for AAC and TAC. However, multivariable model adjustment for other cardiovascular risk factors (BMI, history of smoking, lipids and comorbidities) reduced estimated associations between testosterone and vascular calcification scores at all sites to statistical nonsignificance. [br]Conclusion: Low circulating testosterone levels in men exhibit an association with increased vascular calcification that is not explained by age alone, but which may be accounted for by their co-occurrence with established cardiovascular risk factors.[br][br]Nothing to Disclose: TGT, SB, SB, A-HN, JM, WJM, RSV, CJO 2012-06-25T11:15:00 Theater B 2012-06-25T00:00:00 1899-12-30T11:15:00 2148 275 1917 OR28-1 OR36-01 Monday 1601 2012


1598 ENDO12L_OR28-2 ORAL SESSION: Male Hypogonadism: Predisposition [amp] Risks (11:15 AM-12:45 PM) Late-Onset Hypogonadism (LOH) and Mortality in European Men Stephen R Pye, Ilpo T Huhtaniemi, Terence W O[apos]Neill, Joseph D Finn, David M Lee, Abdelouahid Tajar, Gyorgy Bartfai, Steven Boonen, Felipe F Casanueva, Gianni Forti, Aleksander Giwercman, Thang S Han, Krzysztof Kula, Fernand Labrie, Michael E Lean, Neil Pendleton, Margus Punab, Dirk Vanderschueren, Frederick C Wu The University of Manchester, Manchester, UK; Imperial College London, London, UK; The University of Manchester, Manchester, UK; The University of Oxford, Oxford, UK; Albert Szent-Gyorgy Medical University, Szeged, Hungary; Katholieke Universiteit Leuven, Leuven, Belgium; Santiago de Compostela University, Santiago de Compostela, Spain; University of Florence, Florence, Italy; University of Lund, Malmo, Sweden; Royal Free and University College Hospital Medical School, London, UK; Medical University of Lodz, Lodz, Poland; Laval University, Quebec City, Canada; University of Glasgow, Glasgow, UK; The University of Manchester, Manchester, UK; United Laboratories of Tartu University Clinics, Tartu, Estonia ; Katholieke Universiteit Leuven, Leuven, Belgium Late-onset hypogonadism (LOH) has been defined as a syndrome in middle-aged and elderly men reporting symptoms in the presence of low testosterone (T). Whilst low T has been shown to be associated with all-cause mortality, less is known about the relationship between LOH and mortality. The aim of this study was to determine the association between LOH and mortality in European men.[br]Men aged between 40 and 79 years were recruited from population registers in 8 European centers for participation in a prospective study of male aging: the European Male Aging Study (EMAS). At baseline, subjects attended for a blood sample and completed questionnaires which included information on sexual function, smoking status and number of comorbidities. Serum total T was measured by gas chromatography mass spectrometry. Free T was calculated from total T, SHBG and albumin levels. All subjects were invited to attend a follow-up assessment and reasons for non-participation, including death, were recorded. LOH was defined as the presence of three sexual symptoms (decreased frequency of morning erections, decreased frequency of sexual thoughts, and erectile dysfunction) in combination with total T [ge] 8 and [lt] 11 nmol/L and free T [lt] 220 pmol/L (moderate LOH) and total T [lt] 8 nmol/L and free T [lt] 220 pmol/L (severe LOH). The relationship between LOH status and subsequent mortality was assessed using Cox proportional hazards model with the results expressed as hazard ratios (HR) and 95% confidence intervals (CI).[br]2599 men (mean age 59.1, standard deviation 10.6 years) were included in the analysis, of whom 147 (5.7%) died during a median of 4.3 years of follow-up. Mean T levels were similar in those who died compared to survivors: (16.56 vs 16.64 nmol/L; p=NS). 55 men (2.1%) were classified as having LOH: 31 moderate and 24 severe. After adjusting for age, center, body mass index, current smoking and number of comorbidities, compared to eugonadal men, those with moderate LOH had a two-fold increase in risk of mortality (HR=2.2; 95% CI=1.0, 4.5) and those with severe LOH had a five-fold increase in risk of mortality (HR=5.0; 95% CI=2.4, 10.3). Compared to eugonadal men, those with T [lt] 8 nmol/L regardless of sexual symptoms had a two-fold increase in risk of mortality (HR=2.1; 95% CI=1.1, 4.0).[br]LOH was associated with an increased risk of all-cause mortality in European men. The magnitude of the association was greater than for those with low T only (irrespective of sexual symptoms).[br][br]Nothing to Disclose: SRP, ITH, TWO, JDF, DML, AT, GB, SB, FFC, GF, AG, TSH, KK, FL, MEL , NP, MP, DV, FCW 2012-06-25T11:30:00 Theater B 2012-06-25T00:00:00 1899-12-30T11:30:00 1897 275 1918 OR28-2 OR36-01 Monday 1602 2012


1599 ENDO12L_OR28-3 ORAL SESSION: Male Hypogonadism: Predisposition [amp] Risks (11:15 AM-12:45 PM) Lifestyle Modification Can Reverse Hypogonadism in Men with Impaired Glucose Tolerance in the Diabetes Prevention Program Andrew A Dwyer, Lisa M Caronia, Hang Lee, David M Nathan, Frances J Hayes Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland; Massachusetts General Hospital, Boston, MA; Massachusetts General Hospital, Boston, MA; Massachusetts General Hospital, Boston, MA; St Vincent[apos]s University Hospital, Dublin, Ireland [bold][underline]Background:[/underline][/bold] Testosterone (T) levels tend to be lower in men with obesity and type 2 diabetes than in age-and weight matched controls. We have previously shown that increasing insulin resistance is associated with a decrease in Leydig cell T secretion (1). However, limited interventional data have addressed causality.[br][bold][underline]Aim:[/underline][/bold] To examine the impact of changes in body weight and insulin sensitivity on serum T levels in men.[br][bold][underline]Subjects:[/underline][/bold] 891 men with impaired glucose tolerance (IGT) from the Diabetes Prevention Program (DPP), on no medications which interfere with T levels, who were randomized to lifestyle modification (n=293), metformin (n=305), or placebo (n=293).[br][bold][underline]Methods:[/underline][/bold] Anthropometric variables (BMI, waist circumference), physical activity (MET hours/week), insulin sensitivity (homeostatic model assessment (HOMA-IR) and reproductive hormone levels (T, LH) were analyzed at baseline (BL) and 12 months.[br][bold][underline]Results:[/underline] [/bold]BL age was 53.9 [plusmn] 0.4 y (mean [plusmn] SEM) and BMI 31.9 [plusmn] 0.2 kg/m [sup]2[/sup][sub].[/sub] Mean T levels were not significantly different among treatment groups at BL and in the group as a whole did not change (407 [plusmn] 5 vs 417 [plusmn] 5 ng/dL, BL vs 12 months). However, men randomized to lifestyle modification had a 15% increase in T levels (417 [plusmn] 8 vs 460 [plusmn] 7 ng/dL, P[lt]0.0001) with no change in LH (3.1 [plusmn] 0.1 vs 3.1 [plusmn] 0.1 IU/L). T levels were unchanged in the other treatment groups. The overall prevalence of hypogonadal T levels ([lt] 300 ng/dL) at BL was 23.7%; with lifestyle modification this decreased from 20.4 to 11.1% (P[lt]0.05), but was unchanged with metformin (24.8 vs 23.8%) and placebo (25.6 vs 24.6%).[br]Reduction in body weight was greater with lifestyle modification than metformin (-7.8 vs -2.8 kg, P[lt]0.0001) as was the decrease in HOMA-IR (7.0 [plusmn] 0.3 to 5.2 [plusmn] 0.2 with lifestyle modification vs 7.2 [plusmn] 0.2 to 6.0 [plusmn] 0.3 with metformin). Change in T levels correlated with changes in body weight (r=-0.32, P[lt]0.0001), waist circumference (r=-0.13, P=0.001) and HOMA-IR (r= -0.13, P[lt]0.0001). There was no relationship between change in T and physical activity.[br][bold][underline]Conclusions:[/underline][/bold] (1) Almost 1 in 4 of this male DPP cohort with IGT has low T levels. (2) Lifestyle modification increases endogenous T levels and reduces the prevalence of hypogonadism by 46%. (3) The absence of an increase in LH suggests that enhanced Leydig cell responsiveness underlies the increase in T. (4) Reduction in body weight appears to play an important beneficial role in improving T levels in this population.[br][br](1) Pitteloud N et al., J Clin Endocrinol Metab 2005;90:2636.[br][br]Sources of Research Support: Career Development Award, American Diabetes Association awarded to FJH.[br][br]Nothing to Disclose: AAD, LMC, HL, DMN, FJH 2012-06-25T11:45:00 Theater B 2012-06-25T00:00:00 1899-12-30T11:45:00 1948 275 1919 OR28-3 OR36-01 Monday 1603 2012


1600 ENDO12L_OR28-4 ORAL SESSION: Male Hypogonadism: Predisposition [amp] Risks (11:15 AM-12:45 PM) The Muenster EXAKT Project: Cardiovascular Risk Factors in Klinefelter Patients and Healthy Controls Michael Zitzmann, Rebecca Bongers, Joerg Gromoll, Frank Tuettelmann, Sabine Kliesch University Clinics, Muenster, Germany; University Clinics, Muenster, Germany [bold]Background [amp] Aim[/bold][br]Klinefelter syndrome (47,XXY; KS) is a very common chromosomal disorder, affecting 1:600 men. Klinefelter men have been described to exhibit clinically relevant metabolic patterns related to a pro-inflammatory status, resulting in a high prevalence of insulin resistance and cardiovascular impairment. Testosterone deficiency in form of primary hypogonadism is a common feature in these men.[br]EXAKT ([bold]E[/bold]pigenetics, [bold]X[/bold]-Chromosomal features and clinical [bold]A[/bold]pplications in [bold]K[/bold]linefelter syndrome [bold]T[/bold]rial) is a Muenster-based prospective project involving Klinefelter patients (n=130), and their parents assessing a wide area of cardiovascular, inflammatory and metabolic factors as well as sex steroids and questionnaires in comparison to age-matched healthy male and female controls (2 x n=50). A broad range of genetic and epigenetic investigations completes the approach.[br]Here, we present first and novel clinical data comparing Klinefelter patients to healthy male controls with regard to cardiovascular and metabolic parameters.[br][bold]Results[/bold][br]KS patients had a higher waist circumference and Body Mass Index in comparison to controls. Further on, decreased insulin sensitivity, higher levels of triglycerides and lipoprotein type a as well as lower concentrations of HDL-cholesterol were found in patients. Levels of high-resolution c-reactive protein were elevated in Klinefelter patients. Consequently, the prevalence of the Metabolic Syndrome according the Harmonized Criteria was markedly higher in Klinefelter men than in controls (52/130 vs 5/50). Corroboratingly, carotid artery intima-media thickness was increased and flow mediated dilatation of the brachial artery was decreased in patients vs controls. These differences were statistically significant. Metabolic disadvantages of patients were further enhanced by low testosterone concentrations and already present in the sub-cohort younger than 40 years.[br][bold]Conclusion[/bold][br]The EXAKT study reveals an unfavorable pattern of cardiovascular risk factors in KS in comparison to healthy male controls. This picture is already present in younger patients and enforced by testosterone deficiency.[br][br]Sources of Research Support: The IZKF M[uuml]nster: CRA03/09; and DFG (Grant No. WI2723/4-1).[br][br]Nothing to Disclose: MZ, RB, JG, FT, SK 2012-06-25T12:00:00 Theater B 2012-06-25T00:00:00 1899-12-30T12:00:00 1520 275 1920 OR28-4 OR36-01 Monday 1604 2012


1601 ENDO12L_OR28-5 ORAL SESSION: Male Hypogonadism: Predisposition [amp] Risks (11:15 AM-12:45 PM) Impact of Testosterone Level on Long-Term Outcomes of Men with COPD and Association with Phenotypic Characteristics in a Longitudinal Study, ECLIPSE Christina Wang, Richard V Clark, Bruce E Miller, Lisa D Edwards, Stephen I Rennard, Ruth M Tal-Singer, Ronald S Swerdloff, Richard Casaburi Harbor-UCLA Medical Center, Los Angeles Biomedical Research Institute, Torrance, CA; GlaxoSmithKline, Research Triangle Park, NC; GlaxoSmithKline, King of Prussia, PA; Nebraska Medical Center, Omaha, NE Male hypogonadism has been shown to be associated with increased morbidity and mortality from cardiovascular and respiratory diseases in diverse groups of older men (1, 2). We investigated the association of total testosterone (T) level in 1296 male COPD patients with outcomes and phenotype from the [ldquo]Evaluation of COPD Longitudinally to Identify Predictive Surrogate Endpoints[rdquo] (ECLIPSE) study in which GOLD II-IV COPD patients were followed without additional intervention for three years. T was measured by LC/MS/MS, and free T by equilibrium dialysis.[br]Median T level was 439 ng/dl (IQR - 257). Low T level (expressed as log transformed values) was not correlated with % predicted FEV[sub]1[/sub] but was strongly correlated with higher body mass index (BMI) (Spearman[apos]s r=-0.47) and lower %predicted total lung capacity (r=0.21), each p[lt]0.001. Weaker but statistically significant (p[lt]0.05) correlations of low T were seen with higher age, shorter 6 minute walk distance and lower emphysema score on CT scan. In univariate analysis, subject death tended to be higher in those with low T levels (OR 0.51, p[lt]0.054); lower T level significantly predicted subject death among those with stage II disease [OR 0.24, p[lt]0.003]. In multivariate linear regression analyses that included age, % predicted FEV[sub]1[/sub], BMI, smoking status and T level, both COPD hospitalization and subject death were predicted by age and % predicted FEV[sub]1[/sub] but T level was not predictive of COPD hospitalization and was only predictive of higher subject death in stage II patients (OR 0.25 p[lt]0.007). Free T levels (median 206 pmol/L, IQR - 124) corroborated the total T findings and did not contribute further to the analyses.[br]In a large cohort of men with COPD, total testosterone level correlated with several COPD phenotypic characteristics. Low testosterone tended to predict higher death rate particularly in subjects with moderate airflow obstruction.[br][br](1) Laughlin GA et al., J Clin Endocrinol Metab 2008; 93:68. (2) Shores MM et al., Arch Intern Med 2006; 166:1660.[br][br]Sources of Research Support: This study was supported by GlaxoSmithKline R[amp]D, NCT00292552.[br][br]Disclosures: RVC: Employee, GlaxoSmithKline. BEM: Employee, GlaxoSmithKline. LDE: Employee, GlaxoSmithKline. RMT-S: Employee, GlaxoSmithKline. Nothing to Disclose: CW, SIR, RSS, RC 2012-06-25T12:15:00 Theater B 2012-06-25T00:00:00 1899-12-30T12:15:00 1266 275 1921 OR28-5 OR36-01 Monday 1605 2012


1602 ENDO12L_OR28-6 ORAL SESSION: Male Hypogonadism: Predisposition [amp] Risks (11:15 AM-12:45 PM) Obstructive Sleep Apnea Is Not an Independent Determinant of Plasma Testosterone Gary Wittert, Sean Martin, Robert Adams, Amy Reynolds, Zumin Shi, Anne Taylor, Andre Araujo, Andrew Vakulin University of Adelaide, Adelaide, Australia; University of South Australia, Adelaide, Australia; New England Research Institutes, Watertown, MA; Repatriation General Hospital, Adelaide, Australia It is generally considered that obstructive sleep apnoea (OSA) lowers testosterone in men, although the data supporting this are relatively limited. We determined the relationship between the presence and severity of sleep disordered breathing and plasma testosterone in a comprehensively characterized community based cohort of men aged over 40yrs (MAILES) from whom fasting morning plasma samples were drawn in 2009-2010 (n=1869). Plasma total testosterone (T) was measured by liquid chromatography mass spectrometry (LC-MS/MS). In 2011, home polysomnography was done in 810 randomly selected men from the cohort using an 8 channel Embletta X100 device. The Apnoea Hypopnea Index (AHI), and absence or presence and severity (mild: AHI 10-20; moderate AHI 21-30; severe: AHI [ge] 30) of OSA were classified according to the International Classification of Sleep Disorders (ICDS-2) from the American Academy of Sleep. After excluding men with pathological conditions or taking medications affecting testosterone, with missing values or using CPAP, 654 men aged 41-85 remained.[br]The effect of OSA severity, or AHI, on T were analysed using generalized linear models controlling for potential confounders (age, BMI, smoking, marital status, presence of depression (self-report), HbA1c and SHBG).[br]The mean ([plusmn]SD) characteristics of the men were: age 59 (10) yrs, T 16.5 (5.4) nmol/L, SHBG 33.1 (13.4) nmol/L, BMI 28.4 (4.2) kg/m2, AHI 14.1 (14). OSA was present in 53.7%, (mild 28.6%, moderate 13.6, and severe 11.5%). A significant inverse relationship between AHI and T (Beta -.118, P=0.002), remained after adjustment for age, smoking, marital status, presence of depression, and HbA1c (Beta-.109, P=0.007), and SHBG (Beta -0.077, P=0.017), but not after additional adjustment for BMI (Beta -0.022, P=0.504). The results using OSA category rather than AHI were similar.[br]These data suggest that obesity, or sleep related factors rather than OSA per se, determine T. This accords with the graded effect of weight loss, but limited effect of CPAP to increase T and highlights the importance of managing obesity effectively, particularly in the context of OSA.[br][br]Sources of Research Support: This study was supported by the National Health and Medical Research Council of Australia (NH[amp]MRC grant 627227).[br][br]Disclosures: RA: Speaker, GlaxoSmithKline. Nothing to Disclose: GW, SM, AR, ZS, AT, AA, AV 2012-06-25T12:30:00 Theater B 2012-06-25T00:00:00 1899-12-30T12:30:00 1324 275 1922 OR28-6 OR36-01 Monday 1606 2012


1603 ENDO12L_OR29-1 ORAL SESSION: What[apos]s New in Diagnosis [amp] Treatment of GH Dysfunction? (11:15 AM-12:45 PM) AEZS-130 (Macimorelin)-Stimulated GH Test: Validation of a Novel Test for the Diagnosis of Adult Growth Hormone Deficiency (AGHD) Jose M Garcia, Ronald S Swerdloff, Christina Wang, Michael Kyle, Mark Kipnes, Beverly MK Biller, David Cook, Kevin CJ Yuen, Vivien Bonert, Adrian Dobs, Mark Molitch, George Merriam MEDVAMC, Baylor College of Medicine, Houston, TX; Harbor-UCLA Medical Center and Los Angeles Biomedical Research Institute, Torrance, CA; Radiant Research, Inc, Chicago, IL; DGD Clinic, San Antonio, TX; Massachusetts General Hospital/Harvard Medical School, Boston, MA; Oregon Health [amp] Science University, Portland, OR; Cedars-Sinai Medical Center, Los Angeles, CA; Johns Hopkins Medical Institutions, Baltimore, MD; Northwestern University, Chicago, IL; VA Puget Sound Health Care System/University of Washington, Seattle and Tacoma, WA In the absence of panhypopituitarism and low serum IGF-I levels, the diagnosis of AGHD requires confirmation with a GH stimulation test. The insulin tolerance test (ITT) is the gold standard, but because it induces hypoglycemia, it is contraindicated in patients with coronary artery disease, seizures, and in the elderly.[br]The growth hormone releasing hormone (GHRH) with L-Arginine (L-ARG) test may be used as a safer GH stimulation test for diagnosing AGHD; but GHRH was removed from the US market in 2008, increasing the need for an alternative to the ITT.[br]AEZS-130 is a novel, orally-active growth hormone secretagogue that causes GH secretion by activating the ghrelin receptor. The objective of this clinical trial was to determine the diagnostic efficacy and safety of AEZS-130 in the diagnosis of AGHD.[br]This multicenter open label study was originally designed as a cross-over trial of oral AEZS-130 (0.5 mg/kg) vs GHRH+L-ARG in AGHD patients and in controls, matched for BMI, estrogen status, gender and age. After 43 AGHD patients and 10 controls had been tested, GHRH became unavailable. The study was completed by testing 10 more AGHD patients and 38 controls with AEZS-130 alone.[br]Of the 53 AGHD subjects enrolled, 52 received AEZS-130, and 50 who had confirmed AGHD prior to study entry were included in this analysis, along with 48 controls. Two AGHD subjects could not be matched due to the combination of young age, high BMI and estrogen use.[br]Mean peak GH levels in AGHD patients and controls following AEZS-130 administration were 2.36 (range 0.03-33) ng/mL and 17.71 (range 10.5-94) ng/mL, respectively. The ROC plot analysis yielded an optimal GH cut-point of 2.7 ng/mL, with 82% sensitivity, 92% specificity and a 13% misclassification rate. Obesity (BMI[gt]30) was present in 58% of cases and controls, and peak GH levels were inversely associated with BMI in controls.[br]Adverse events (AE) were seen in 19 of 52 AGHD patients (37%) and in 10 of 48 (21%) controls following AEZS-130. In contrast, 26 of 43 (61%) AGHD subjects and 3 out of 10 (30%) controls experienced AEs with L-ARG+GHRH. Adverse events were generally mild or moderate in severity. Only one drug-related serious AE (SAE) was reported in a control subject receiving AEZS-130 (2.1%), whose ECG post-dosing showed QT prolongation and non-specific T wave abnormalities that were asymptomatic and resolved spontaneously within 24 hours. In conclusion, this study showed that AEZS-130 is safe and effective in diagnosing AGHD.[br][br]Sources of Research Support: This study was funded by AEterna Zentaris GmbH.[br][br]Disclosures: JMG: Investigator, Aeterna Zentaris Inc. RSS: Investigator, Aeterna-Zentaris. MK: Investigator, Aeterna-Zentaris. MK: Investigator, Aeterna-Zentaris. BMKB: Investigator, Æterna Zentaris GmbH; Consultant, Æterna Zentaris GmbH. DC: Investigator, Æterna Zentaris GmbH. KCJY: Investigator, Æterna Zentaris GmbH. VB: Investigator, Aeterna-Zentaris. AD: Investigator, Aeterna-Zentaris. MM: Investigator, Aeterna Zentaris, Inc, Novo Nordisk, Eli Lilly & Company. GM: Investigator, Aeterna-Zentaris. Nothing to Disclose: CW 2012-06-25T11:15:00 Theater A 2012-06-25T00:00:00 1899-12-30T11:15:00 584 276 1923 OR29-1 OR21-01 Monday 1607 2012


1604 ENDO12L_OR29-2 ORAL SESSION: What[apos]s New in Diagnosis [amp] Treatment of GH Dysfunction? (11:15 AM-12:45 PM) Audit of Pituitary Dysfunction after Traumatic Brain Injury: Caution in Interpretation of Glucagon Stimulation Test in Diagnosis of GH and ACTH Deficiency Carmen Tenorio Jimenez, Mari Niemik, Aysha Malik, Debbie Papadopoulou, Timothy E Ham, David Baxter, David J Sharp, Tony Goldstone Imperial College Healthcare NHS Trust, London, UK; Imperial College London Hammersmith Campus, London, UK; Imperial College London Hammersmith Hospital, London, UK [bold]Introduction.[/bold] Pituitary dysfunction after TBI has received increasing recognition in recent years because of its potential contribution to morbidity. Differences in diagnostic endocrine tests and diagnostic criteria may explain the variation in the prevalence of pituitary dysfunction between studies. [bold]Objectives[/bold]. Audit of the prevalence of pituitary dysfunction, and the validity of the glucagon stimulation test (GST) as a diagnostic tool for GH and ACTH deficiency after TBI. [bold]Patients and Methods.[/bold] 191 patients (143 males), mean [plusmn] SD age 40.7 [plusmn] 15.4yr attended initial assessment in the TBI outpatient clinic between July 2009 and Aug 2011. Median time since TBI was 0.27yr (74.8% [lt]1yr). 22% had an absolute contraindication for insulin tolerance testing (ITT) and 38.7% had a relative contraindication. Patients had basal pituitary function tests and 150 (78.6%) had a GST for diagnosis of GH and ACTH deficiency. Where clinically appropriate, patients had a GHRH-Arginine test for confirmation of GH deficiency, a Metyrapone suppression test (MST) for confirmation of ACTH deficiency, and a water deprivation test (WDT) for confirmation of diabetes insipidus. [bold]Results.[/bold] There were no patients with TSH deficiency or diabetes insipidus, but one patient had syndrome of inappropriate ADH. 2/183 had asymptomatic hyperprolactinaemia and 3/185 had gonadotrophin deficiency. 36/150 had a GST peak cortisol [lt]350 nmol/L (24.0%), but only 1/25 (4%) had confirmation of ACTH deficiency by failing a MST. None of the 16 patients with GST peak cortisol [gt]350 nmol/L failed a MST. 39/150 had a GST peak GH [lt]3 mcg/L (26.0%) and 63/150 peak GH [lt]5 mcg/L (42.0%), but only 3/30 (10%) had GH deficiency confirmed by failing a GHRH-Arginine test. Patients with confirmed GH deficiency had worse ratings of Quality of Life Questionnaires and have been commenced on GH replacement with symptomatic improvement. [bold]Conclusions.[/bold] The GST had a high false positive rate in diagnosing both GH and ACTH deficiency in patients after TBI. This is likely related to the low prevalence of true pituitary dysfunction in this group. However detection of pituitary dysfunction after TBI remains clinically important, while GH replacement is beneficial. Despite the high prevalence of contraindications to an ITT and associated resource implications, caution must be made not to over-diagnose GH and particularly ACTH deficiency after TBI by relying on a GST alone and second line confirmatory tests must be performed for both axes.[br][br]Nothing to Disclose: CTJ, MN, AM, DP, TEH, DB, DJS, TG 2012-06-25T11:30:00 Theater A 2012-06-25T00:00:00 1899-12-30T11:30:00 2308 276 1924 OR29-2 OR21-01 Monday 1608 2012


1605 ENDO12L_OR29-3 ORAL SESSION: What[apos]s New in Diagnosis [amp] Treatment of GH Dysfunction? (11:15 AM-12:45 PM) A Phase 1 Trial of Safety, Pharmacokinetics (PK) and Pharmacodynamics (PD) of a Single Dose of a New Human Growth Hormone Analogue (VRS-317) for Monthly Subcutaneous Administration in Adults with Growth Hormone Deficiency Timothy Bailey, BMK Biller, Gerard S Conway, Mark Kipnes, Vera Popova-Brkic, Kevin CJ Yuen, Eric Humphriss, George M Bright, Jeffrey L Cleland AMCR Institute, Escondido, CA; Massachusetts General Hospital, Boston, MA; University College London Hospitals, London, UK; DGD Clinic, San Antonio, TX; University Clinic Center, Belgrade, Serbia; Oregon Health Sciences University, Portland, OR; Versartis, Inc, Redwood City, CA; Versartis, Inc, Redwood City, CA; Versartis, Inc, Redwood City, CA VRS-317, a novel long acting rhGH fusion protein, is under study in 50 adults with GHD in a 60-day, double-blind, randomized, placebo(PBO)-controlled, single ascending dose study of 0.05, 0.10, 0.20, 0.40 and 0.80 mg VRS-317/kg (ClinicalTrials.gov NCT01359488). Preliminary results from an ongoing trial are reported here. VRS-317 is [sim] 5 times the mass of rhGH due to the addition of N- and C-terminal amino acids to extend the rhGH half-life. In monkeys, VRS-317 has complete bioavailability, rapid absorption, a half-life of [sim]110 hr, and produces a sustained IGF-I response for one month after a single dose.[br]Initially, subjects took daily rhGH (min. 28 days, dose range 0.2-1.2 mg/d) until serum IGF-1 SD score was stable in the range of -1.5 and +1.5 and then withdrawn from hGH until the IGF-I SDS was [lt] -1 and had fallen by [ge] 0.75. The subjects were observed for 48 hrs upon receiveingVRS-317 or PBO. PK, PD (IGF-I) and paired fasting/post-prandial glucose were measured pre-dose and at various times over 30 days after a single SC dose of VRS-317 or PBO. At abstract submission, safety data were available for 28 subjects and PK/PD for 24 VRS-317 or PBO-treated subjects. PK/PD subjects (15M, 9F) had a mean (SD) age of 46 (12) yrs and BMI of 32 (7) kg/m[sup]2[/sup].[br]VRS-317 achieves a Tmax 2-3 days after a SC dose and has a long circulating half-life potentially sufficient for monthly dosing. The mean maximal increases in IGF-I SDS were 0.33, 0.32, 0.96* and 1.32** in the PBO, 0.05, 0.10 and 0.20 mg/kg/month dosing groups, respectively (*p = 0.012, ** p = 0.0005 (vs.PBO)). The percentages of subjects with IGF-I SDS above pre-VRS-317 levels for the initial two weeks were 16, 66 and 100% for the 0.05, 0.10 and 0.20 mg/kg/month groups, respectively. These single VRS-317 doses are equivalent to 0.33, 0.66 and 1.32 [mu]g hGH/kg/day (typical AGHD dosing range for daily rhGH is 2-12 [micro]g/kg/d).[br]There were no drug-related serious adverse events, withdrawals after dosing or unexpected, related adverse events or injection site lipoatrophy. Mean fasting glucose, post-prandial glucose and change from fasting to postprandial showed no significant post-dosing changes. No safety laboratory signals were observed.[br]In summary, in this trial of a single SC dose of VRS-317 in adults with GHD, graded responses of IGF-1 generation were safely achieved at doses lower than those typically used with daily rhGH over the course of one month.[br][br]Disclosures: MK: Investigator, Aeterna-Zentaris. EH: Employee, Versartis, Inc. GMB: Employee, Versartis, Inc. JLC: Chairman, Versartis, Inc. Nothing to Disclose: TB, BMKB, GSC, VP-B, KCJY 2012-06-25T11:45:00 Theater A 2012-06-25T00:00:00 1899-12-30T11:45:00 631 276 1925 OR29-3 OR21-01 Monday 1609 2012


1606 ENDO12L_OR29-4 ORAL SESSION: What[apos]s New in Diagnosis [amp] Treatment of GH Dysfunction? (11:15 AM-12:45 PM) A Phase 2, Multiple-Dose, Open-Label, Parallel-Group, Active Controlled, Safety, Tolerability, Pharmacokinetic and Pharmacodynamic Study of ACP-001 in Adult Patients with Growth Hormone Deficiency (AGHD) Charlotte Hoybye, Andreas FH Pfeiffer, Diego Ferone, David Gilfoyle, Michael Beckert, Jens Sandahl Christiansen Karolinska University Hospital, Stockholm, Sweden; Charit[eacute] University Hospital, Berlin, Germany; San Marino University Hospital, Genova, Italy; Ascendis Pharma A/S, Hellerup, Denmark; Aarhus University Hospital, Aarhus C, Denmark ACP-001 is designed as a long-acting pro-drug that releases unmodified human Growth Hormone (hGH). The carrier-linked pro-drug is absorbed into the blood compartment, where it acts as a circulating hGH reservoir, releasing the hGH over a defined period of time. In a previous Phase 1 study, ACP-001 was safe and well tolerated and demonstrated pharmacodynamic effects (IGF-I) at least comparable to daily hGH injections.[br]Objectives[br]The objectives of this study were to investigate 1) safety and tolerability of ACP-001 under multiple-dose conditions in adult patients with Growth Hormone Deficiency and 2) pharmacokinetics (PK) and pharmacodynamics (PD) over a 4-week period, in comparison to daily hGH.[br]Method[br]Study population: 37 male and female patients with verified Growth Hormone Deficiency and hGH replacement therapy for at least 3 months. Following a wash-out period, patients were randomized to one of three dose-levels of ACP-001 (equivalent to 0.02, 0.04 and 0.08 mg hGH/kg/week) injected weekly, or to daily hGH (0.04 mg/kg/week) over 28 days.[br]Safety, immunogenicity, pharmacodynamic and pharmacokinetic parameter samples were taken throughout the entire period, with full PK and PD in week one and week four and a follow-up at Day 42.[br]Results[br]All dose-levels of ACP-001 were safe and well tolerated. Most frequent AEs were headache and fatigue. Two SAEs occurred in one patient, judged as not drug-related. A total of nine patients experienced injection site reactions, the most frequent being mild erythema and reported across all treatment groups. No lipoatrophy at the injection site was observed. No treatment emergent antibodies occurred. Data showed PK and PD dose-response of the different dose levels of ACP-001. IGF-I levels increased in a dose-dependent manner and demonstrated a similar response for ACP-001 0.04 mg/kg/week compared to the corresponding dose of daily hGH (0.04 mg/kg/week).[br]Conclusion[br]ACP-001 was safe and well tolerated in this Phase 2 study in adult patients with Growth Hormone Deficiency. Pharmacokinetic results support a once-weekly dosing. IGF-I increased in a dose-dependent manner and comparable to daily hGH, indicating that ACP-001 is at least as potent as the corresponding dose of daily hGH.[br][br]Disclosures: DG: Employee, Ascendis Pharma A/S. MB: Consultant, Ascendis Pharma A/S. Nothing to Disclose: CH, AFHP, DF, JSC 2012-06-25T12:00:00 Theater A 2012-06-25T00:00:00 1899-12-30T12:00:00 633 276 1926 OR29-4 OR21-01 Monday 1610 2012


1607 ENDO12L_OR29-5 ORAL SESSION: What[apos]s New in Diagnosis [amp] Treatment of GH Dysfunction? (11:15 AM-12:45 PM) Once-Weekly, CTP-Modified hGH (MOD-4023) Is Effective in Growth Hormone Deficient Adults: A Phase II, Dose and Frequency Finding Study Vera Popovic, Miklos I Goth, Peter Vanuga, Juraj Payer, Marija Pfeifer, Martin Bidlingmaier, Eyal Udi Fima University Clinical Center, Belgrade, Serbia; University of Belgrade, Belgrade, Serbia; Military Hospital, Budapest, Hungary; National Institute of Endocrinology and Diabetology, Lubochna, Slovakia (Slovak Republic); Comenius University, Bratislava, Slovakia (Slovak Republic); University Medical Centre Ljubljana, Ljubljana, Slovenia; Medizinische Klinik Campus Innenstadt, Munich, Germany; Prolor-Biotech, Ltd, Nes Ziona, Israel [italic][bold]Objective:[/bold] [/italic]Growth Hormone (GH) replacement therapy currently requires daily injections, which may cause poor compliance, inconvenience and distress for patients. CTP-modified hGH (MOD-4023) is being developed for once-weekly administration in Growth Hormone Deficient (GHD) adults and children. The present study evaluated the safety, tolerability, pharmacokinetics (PK) and pharmacodynamics (PD) of three doses of MOD-4023 in GHD adults.[br][italic][bold]Design and methods:[/bold] [/italic]39 GHDA patients (33 males and 6 females) who had IGF-1 levels within [plusmn]2 SDS during daily hGH replacement were randomized and switched to MOD-4023 administered as once-weekly subcutaneous injections to evaluate safety, tolerability and PK/PD profile with 3 doses. The doses were 30%, 45% or 100% of each patient[apos]s cumulative weekly hGH dose. The study was comprised of two stages. Stage I included an optimization period and 4 weeks of once-weekly MOD-4023. Stage II is an optional 16 week extension period of once weekly MOD-4023 administration to collect further safety information and confirm the results obtained in Stage I. Here we present the results of Stage I.[br][italic][bold]Results:[/bold] [/italic]MOD-4023 was well-tolerated and demonstrated efficacy at all 3 doses. MOD-4023 treatment resulted in a dose-dependent response of maximal IGF-1 concentration. In the majority of patients from all cohorts, IGF-I levels were maintained within [plusmn]2 SDS during the four weeks of treatment with MOD-4023. IGF-1 levels did not exceed the upper limit of +2 SDS in any patients. In two treatment cohorts (45% and 100% of the weekly cumulative hGH dose), the mean IGF-1 values were comparable to those obtained with daily hGH at steady state. No drug-related Serious Adverse Events were reported during the study. The adverse effects reported (mainly headaches) were consistent with known hGH related side-effects, and were mostly mild. MOD-4023 was not immunogenic.[br][italic][bold]Conclusions:[/bold] [/italic]Once-weekly, repeated doses of long-acting MOD-4023 were shown to be safe and well tolerated in adult GHD patients. IGF-I levels were maintained within the normal range in most MOD-4023 treated patients for the entire 4 weeks.[br]Based on the positive results of Stage I, an estimated target dose range for the subsequent Phase III study has been established and the 16-week extension study was initiated to further support the estimated dose range and to provide additional safety information.[br][br]Disclosures: MB: Medical Advisory Board Member, Prolor-Biotech Ltd. EUF: Employee, Prolor-Biotech, Ltd. Nothing to Disclose: VP, MIG, PV, JP, MP 2012-06-25T12:15:00 Theater A 2012-06-25T00:00:00 1899-12-30T12:15:00 1177 276 1927 OR29-5 OR21-01 Monday 1611 2012


1608 ENDO12L_OR29-6 ORAL SESSION: What[apos]s New in Diagnosis [amp] Treatment of GH Dysfunction? (11:15 AM-12:45 PM) Pharmacokinetic Modeling of Oral Octreotide (Octreolin[trade]) in Healthy Volunteers and Dosing Regimen Optimization for Acromegaly Patients Shmuel Tuvia, William G Kramer, Dori Pelled, Sam L Teichman, David L Kleinberg, Shlomo Melmed, Roni Mamluk Chiasma, Jerusalem, Israel; Kramer Consulting, North Potomac, MD; New York University, New York, NY; Cedars Sinai Medical Center, Los Angeles, CA [bold]Background:[/bold] Somatostatin analogs including octreotide are the first-choice pharmacotherapy for treating acromegaly patients. Octreotide achieving a threshold level after parenteral administration results in suppression of GH levels leading to reduced IGF-1 levels. We have developed an oral formulation of unmodified octreotide (Octreolin) using the Transient Permeability Enhancer technology. Octreolin pharmacokinetic[apos]s (PK) were compared to subcutaneous (SC) therapeutic dosing of injected octreotide in healthy volunteers (HV).[br][bold]Objective: [/bold]To design an Octreolin dose regimen for acromegaly patients using PK results in HV.[br][bold]Methods: [/bold]Octreotide plasma concentrations were measured over 12-24 hours after a single oral or SC dose of octreotide to 74 HV. Time frames for octreotide therapeutic concentrations ([ge] 0.5 ng/mL and [ge] 1 ng/mL) were also estimated. Estimates and variability of these metrics after oral or SC octreotide were assessed using a bootstrap analysis.[br][bold]Results:[/bold] Plasma octreotide concentrations after 3, 10 and 20 mg Octreolin were proportional to the dose [AUC[sub]inf[/sub] (2.0[plusmn]0.3 h[times]ng/mL, 8.0[plusmn]0.8 h[times]ng/mL and 15.0[plusmn]4.2 h[times]ng/mL, respectively)]. Octreolin capsule (20 mg) or SC injection (0.1 mg) showed comparable AUC[sub]inf[/sub](16.2[plusmn]1.25 h[times]ng/mL, 12.1[plusmn]0.45 h[times]ng/mL, respectively) and Cmax (3.77[plusmn]0.25 ng/mL, 3.97[plusmn]0.19 ng/mL, respectively), but the plasma octreotide concentrations increased more slowly after oral than SC (median Tmax of 2.7 h and 0.6 h, respectively). Plasma octreotide concentrations remained above the two therapeutic thresholds for 7.7 hours and 5.9 hours (range, 0.67[ndash]23.7 h and 3.92[ndash]7.92 h, respectively) after Octreolin dose and for 6.0 hours and 3.9 hours (range, 0.33[ndash]11.7 h and 2.67[ndash] 4.92 h, respectively) after the injectable administration. Bootstrap analysis suggested that 20 mg oral octreotide given [italic]b.i.d[/italic] every 12 hours should be comparable to 0.1 mg octreotide SC injection administered [italic]t.i.d[/italic]. (every 8 hours), with respect to maintaining octreotide concentrations above the therapeutic threshold. Consequently, a dose regimen of Octreolin 20 mg [italic]b.i.d[/italic] with an option for dose escalation would be feasible for testing effects on GH and IGF-1 in acromegaly patients[italic].[/italic][br][bold]Conclusions[/bold]: Systemic exposure to orally administrated octreotide was similar to that of a therapeutic dose of SC injection in HV. Bootstrap analysis indicated a comparable time with concentrations above the therapeutic threshold for [italic]b.i.d. [/italic]dosing of oral octreotide and for parenteral octreotide.[br][br]Disclosures: ST: Employee, Chiasma, Inc. WGK: Consultant, Chiasma. DP: Employee, Chiasma. SLT: Employee, Chiasma. DLK: Medical Advisory Board Member, Novartis Pharmaceuticals; Researcher, Novartis Pharmaceuticals; Principal Investigator, Novartis Pharmaceuticals. SM: Medical Advisory Board Member, Chiasma. RM: Employee, Chiasma. 2012-06-25T12:30:00 Theater A 2012-06-25T00:00:00 1899-12-30T12:30:00 1380 276 1928 OR29-6 OR21-01 Monday 1612 2012


1609 ENDO12L_OR30-1 ORAL SESSION: Growth [amp] Puberty (11:15 AM-12:45 PM) Baseline Gene Expression Associated with Growth Response to Growth Hormone (GH) in Children with GH Deficiency (GHD): The PREDICT Long-Term Follow-Up Study Adam Stevens, Regis Coutant, Gerhard Binder, Benoit Destenaves, Pierre Chatelain, Peter Clayton University of Manchester, Manchester, UK; La Trevaresse Che Fontrousse, Aix en Provence, France; CHU d[apos]Angers, H[ocirc]pital Robert Debr[eacute], Angers, France; University Children[apos]s Hospital, Tuebingen, Germany; Merck Serono SA, Geneva, Switzerland; Universit[eacute] Claude Bernard, Bron, France; University of Manchester, Manchester, UK Background: The PREDICT long-term follow-up study investigates relationships between conventional biomarkers, genetic polymorphisms, gene expression, and long-term auxological changes in GH treatment-na[iuml]ve prepubertal children with GHD during GH therapy.[br]Objective: To investigate the association between baseline gene expression and Year 1 growth response to GH therapy in children with GHD.[br]Patients: Prepubertal, treatment-na[iuml]ve children with classic idiopathic GHD.[br]Methods: Height velocity (HV) after 1 year of GH therapy (median dose 33 [micro]g/kg/day) was categorized by quartile and correlated with basal gene expression (W0), measured using the Affymetrix Human Genome U133 Plus 2.0 Array. Analysis of variance (ANOVA) corrected for false discovery rate (q-value) was used to determine differential gene expression between quartiles. Gene ontology was assessed using the hypergeometric statistical test. Biological network inference was performed using Ingenuity Pathway Analysis software, along with Agilent Literature Search and Reactome Functional Interaction Cytoscape Plug-ins. Predictive modeling was performed using K-nearest neighbor classification, with 1-level cross-validation and variables selected by shrinking centroids.[br]Results: In the W0 samples (n=71), differences in the expression of 526 probes correlated with the lowest quartile of HV (ANOVA, q[lt]0.2). All differences in expression were [gt]1.1-fold (range [ndash]2.2 to +1.5). Gene ontology and inferred pathway analysis identified multiple networks of genes associated with an enrichment of metabolic functions; the cell cycle; and pathways related to apoptosis, cardiovascular system development, and growth (q[lt]0.05). Changes in gene expression within the insulin growth factor-1 pathway were correlated with the lowest quartile of HV after 1 year (p[lt]0.05), including [italic]FOXO3[/italic] (+1.3-fold), [italic]JAK1 [/italic](+1.2-fold), [italic]NRAS[/italic] ([ndash]1.2-fold), and [italic]CSNK2A1[/italic] (+1.2-fold). Predictive modeling identified a group comprising 26 gene-expression probes, which had a positive predictive value for poor growth responders of 0.48 and a negative predictive value of 0.86.[br]Conclusions: Using baseline gene-expression profiles, this work has identified candidate biological pathways associated with low growth response over the first year of GH therapy in prepubertal children with GHD. Baseline gene-expression profiles may add value to a predictive model for individualized GH treatment based on auxological parameters, serum biomarkers, and genetic polymorphisms.[br][br]Sources of Research Support: Merck Serono S.A. [ndash] Geneva, Switzerland, a branch of Merck Serono S.A., Coinsins, Switzerland, an affiliate of Merck KGaA, Darmstadt, Germany.[br][br]Disclosures: GB: Investigator, Novo Nordis, Pfizer, Inc.; Speaker, Eli Lilly & Company, Ferring Pharmaceuticals, Ipsen, Novo Nordisk, Pfizer, Inc. BD: Employee, Merck Serono S.A. PC: Investigator, Merck Serono S.A.; Consultant, Merck Serono S.A.; Speaker, Merck Serono S.A. PC: Investigator, Merck Serono S.A.; Speaker, Merck Serono S.A. Nothing to Disclose: AS, RC 2012-06-25T11:15:00 362 2012-06-25T00:00:00 1899-12-30T11:15:00 605 277 1929 OR30-1 OR29-01 Monday 1613 2012


1610 ENDO12L_OR30-2 ORAL SESSION: Growth [amp] Puberty (11:15 AM-12:45 PM) Incidence of Pathology Detection and Cost of Screening in the Evaluation of Children with Short Stature Stephanie R Sisley, Marcela Vargas Trujillo, Philippe Backeljauw Cincinnati Children[apos]s Hospital Medical Center, Cincinnati, OH; Cincinnati Children[apos]s Hospital Medical Center, Cincinnati, OH [bold]Background:[/bold] The idiopathic short stature (ISS) consensus statement recommends extensive screening laboratory testing in short patients for whom history and exam do not suggest a specific diagnosis (1). However, no study has analyzed the clinical utility and cost-effectiveness of such comprehensive screening in otherwise asymptomatic short children.[br][bold]Aims:[/bold] 1) Determine the incidence of pathology found during routine screening of otherwise healthy short children. 2) Determine adherence to the ISS consensus statement diagnostic guidelines. 3) Determine the cost per identified diagnosis from comprehensive screening evaluation.[br][bold]Methods:[/bold] Retrospective review of 1,373 short stature referrals to a pediatric endocrinology clinic between 2008 and 2011. Exclusion criteria: 1) height [gt] 3[sup]rd[/sup] %-ile, 2) prior height velocity [lt] 5cm/yr, or 3) findings on history or exam warranting a specific work up. 250 subjects were included for analysis. Age at presentation, gender, parental heights, 12-point review of systems, physical exam, testing results (CBC, ESR, BMP, TSH, Free T4, T4, IGF-1, IGFBP-3, TTG, IgA, chromosomes, urinalysis, bone age), and working diagnosis were collected.[br][bold]Results: [/bold]72% of patients were male with a mean age of 9.97[plusmn]4.32 yrs; 58% were pre-pubertal. Mean height SDS was -2.52[plusmn]0.52, mean weight SDS was -2.25[plusmn]1.14, and mean BMI was -0.82[plusmn]1.3 SDS. Pre-evaluation height velocity averaged 6.5 cm/yr (n=101 with growth records). Most represented normal variants of growth; 6% were born SGA (15/250). 18% (46/250) were labeled ISS with normal test results. The incidence of newly diagnosed pathology was 1.2% (n=3/250). One patient was diagnosed with celiac disease. Two other subjects had abnormal test results, but neither was confirmed due to lack of follow-up. One patient had elevated TTG and one had increased IGF-1, respectively indicative of celiac disease and an IGF-1 receptor defect. Only 9.2% of the patients had all tests done as recommended in the ISS consensus paper. To test the 250 patients, the cost was $344,244, yielding $114,748 per diagnosis. If the full ISS consensus statement testing guidelines had been adhered to, the cost would have been $147,340/new diagnosis.[br][bold]Conclusions: [/bold]The incidence of new disease found during routine, comprehensive screening in asymptomatic short children is very low. We suggest that healthy short children do not warrant non-directed, comprehensive testing and that future guidelines should include patient-specific testing.[br][br](1) Cohen P, et al. J Clin Endocrinol Metab 2008; 93(11):4210-4217.[br][br]Sources of Research Support: NIH Grant 5T32ES010957-10 awarded to SRS; NIH Grant 1F32DK091077-01A1 awarded to SRS.[br][br]Nothing to Disclose: SRS, MVT, PB 2012-06-25T11:30:00 362 2012-06-25T00:00:00 1899-12-30T11:30:00 1843 277 1930 OR30-2 OR29-01 Monday 1614 2012


1611 ENDO12L_OR30-3 ORAL SESSION: Growth [amp] Puberty (11:15 AM-12:45 PM) Intranasal Administration of Human Growth Hormone (CP024) and Induction of IGF-1 in Healthy Volunteers Faron Michael Jordan, Lisbeth Illum, Stephen Shalet, Gareth King, Andrew Lester Lewis Critical Pharmaceuticals, Nottingham, UK; Christies Hospital, Manchester, UK; BioCity Nottingham, Nottingham, UK [bold]Rationale[/bold][br]Patient adherence to growth hormone replacement therapy is estimated to be as low as 36-49%[sup]1[/sup] with 70% of patients unhappy with daily injection and 30% considering stopping treatment (Frost [amp] Sullivan). Efficacy is related to treatment adherence (Acerin 2007) and consequently suboptimal adherence reduces efficacy and increases healthcare costs. All of the presently available marketed formulations of growth hormone require subcutaneous injection and an intranasally delivered formulation that increases patient convenience and ease of use should increase adherence and treatment success.[br][bold]Nasal Human Growth Hormone Formulations[/bold][br]CriticalSorb[trade] is an advanced nasal delivery system that facilitates the absorption of macromolecules across biological membranes. We have used this technology to develop CP024, a nasal spray formulation of human growth hormone (hGH), able to achieve bioavailabilities of 20-43% relative to subcutaneous injection in rats, rabbits and cynomolgus monkeys, which is higher than any previously reported[sup]2,3[/sup]. These formulations have been shown to be well tolerated in a 14 day repeat intranasal dose toxicity study. CriticalSorb[trade] itself is already used in a number of marketed products for oral and intravenous administration and was shown to be well tolerated in a six month repeat intranasal dose toxicity study. Furthermore, the nasal application of growth hormone provides peak plasma levels over a period of 2-3 hours similar to that expected with endogenous growth hormone secretion in children. A Phase 1 study was carried out to evaluate the bioavailability and bioactivity of two CP024 formulations administered intranasally to healthy volunteers relative to a subcutaneous injection of Omnitrope[reg].[br][bold]Trial Design and Results[/bold][br]Single centre, open label, five-way crossover in eight healthy volunteers. Endogenous GH secretion was suppressed by an infusion of octreotide.[br]The results showed that CriticalSorb significantly and reproducibly enhanced the absorption of hGH across the nasal mucosa after intranasal administration. The formulations were also well tolerated with highly reproducible pharmacokinetics. More importantly however, IGF-1 was strongly induced and the induction was similar to that seen after a subcutaneous injection of Omnitrope[reg]. This is the first report of IGF-1 induction following intranasal administration of hGH[sup]4[/sup]. A second trial is underway to investigate the dose response relationship for CP024 and IGF-1 induction.[br][br]1. F. Haverkamp, L. Johansson, H. Dumas, S. Langham, M. Tauber, D. Veimo, F. Chiarelli. Observations of nonadherence to recombinant human growth hormone therapy in clinical practice. Clin Ther, 30(2) (2008) 307-316. 2. H.R. Costantino, L. Illum, G. Brandt, P.H. Johnson, S.C. Quay, Intranasal delivery: Physicochemical and therapeutic aspects, Int. J. Pharm. 337 (2007) 1-24. 3. Y.H. Cheng, A.M. Dyer, I. Jabbal-Gill, M. Hinchcliffe, R. Nankervis, A. Smith, P. Watts. Intranasal delivery of recombinant human growth hormone (somatropin) in sheep using chitosan-based powder formulations. Eur J Pharm Sci. 2005 Sep;26(1):9-15. 4. T. Laursen, B. Grandjean, J.O.L. J[oslash]rgensen, J.S. Christiansen. Bioavailability and bioactivity of three different doses of nasal growth hormone (GH) administered to GH-deficient patients: comparison with intravenous and subcutaneous administration. Eur J Endocrinol (1996) 135: 309-15.[br][br]Sources of Research Support: Critical Pharmaceuticals, The Wellcome Trust.[br][br]Nothing to Disclose: FMJ, LI, SS, GK, ALL 2012-06-25T11:45:00 362 2012-06-25T00:00:00 1899-12-30T11:45:00 1559 277 1931 OR30-3 OR29-01 Monday 1615 2012


1612 ENDO12L_OR30-4 ORAL SESSION: Growth [amp] Puberty (11:15 AM-12:45 PM) A Large Number of Genes Involved in Growth Plate Chondrogenesis Contribute to Human Height Variation Julian C Lui, Ola Nilsson, Yingleong Chan, Cameron D Palmer, Anenisia C Andrade, Joel N Hirschhorn, Jeffrey Baron National Institute of Child Health and Human Development, Bethesda, MD; Karolinska Institutet and Karolinska University Hospital, Stockholm, Sweden; Boston Childen[apos]s Hospital, Boston, MI; Broad Institute, Cambridge, MI; Harvard Medical School, Boston, MI Previous meta-analysis of genome-wide association studies (GWAS) has identified 180 loci that influence adult height (Nature 467: 832-8, 2010). However, as with most GWAS, the data typically do not pinpoint a single gene at each locus but instead identify a set of contiguous genes, only one of which presumably modulates height.[br]We reasoned that many of the causative genes within these loci influence height because they are expressed in and function in the growth plate. We therefore used microarray analysis in mice and rats to identify [ldquo]GP-array genes[rdquo] within these loci that show 1) high expression in growth plate cartilage compared to other tissues; 2) expression that is spatially regulated across different growth plate zones, suggesting a role in chondrocyte differentiation; or 3) expression that is temporally regulated in the growth plate, suggesting a role in growth plate senescence. We found that the GWAS loci were strongly enriched in these GP-array genes (8.5% in GWAS loci vs 3.2% in the whole genome, P [lt] 0.0001) and that the GWAS genes that lie closest to the height-associated SNPs showed particularly strong enrichment (15.7%, P [lt] 0.0001), validating this approach. To identify additional growth plate genes from the GWAS list, we used bioinformatic analysis to find genes for which mutations cause a growth plate phenotype in mice or humans. The genes in the GWAS list, and particularly those genes closest to the height-associated SNPs, showed strong enrichment for genes with a human and mouse growth plate phenotype (both P [lt] 0.0001), validating this bioinformatic approach. Together, these expression and functional approaches implicated 78 genes from the GWAS loci, including multiple genes involved in the IHH-PTHrP negative feedback loop, BMP signaling, C-type natriuretic peptide signaling, and GH-IGF1 signaling. Many identified genes have not previously been implicated in human growth plate regulation.[br]In summary, our analysis provides strong evidence that human height variation is due in large part to genes that act locally in the growth plate to influence chondrogenesis. These genes are strong candidates for causing skeletal dysplasias, syndromic short stature, and isolated short stature of unknown etiology. The identified genes also suggest new regulatory pathways in the growth plate, inviting functional investigation.[br][br]Nothing to Disclose: JCL, ON, YC, CDP, ACA, JNH, JB 2012-06-25T12:00:00 362 2012-06-25T00:00:00 1899-12-30T12:00:00 835 277 1932 OR30-4 OR29-01 Monday 1616 2012


1613 ENDO12L_OR30-5 ORAL SESSION: Growth [amp] Puberty (11:15 AM-12:45 PM) A Loss-of-Function Mutation in [italic]TACR3 [/italic]Gene in a Patient with Gonadotropin-Dependent Precocious Puberty Ana Claudia S Reis, Noel D Sekoni, Karla M Ishikawa, Carla Y Tarumi, Carlos E Martinelli, Jr, Margaret Castro, Rona S Carroll, Ursula B Kaiser, Sonir R Antonini School of Medicine of Rebeir[atilde]o Preto - University of S[atilde]o Paulo, Rebeir[atilde]o Preto, Brazil; School of Medicine of Rebeir[atilde]o Preto - University of S[atilde]o Paulo, Rebeir[atilde]o Preto, Brazil; Brigham and Women[apos]s Hospital and Harvard Medical School, Boston, MA [bold]Introduction:[/bold] Whether idiopathic gonadotropin-dependent precocious puberty (GDPP) is a monogenic disorder remains elusive. To date, mutations in genes involved in GnRH regulation were found in only two GDPP patients, harboring [italic]KISS1[/italic] and [italic]KISS1R[/italic] mutations. Neurokinin B (NKB) has recently emerged as a new regulator of GnRH secretion, co-expressed with kisspeptin in the hypothalamus. Inactivating mutations in NKB and in its receptor (NK3R) cause hypogonadotropic hypogonadism. Interestingly, animal studies suggest both inhibitory and stimulatory effects of NKB on GnRH secretion, depending on the steroid hormone milieu. We investigated the presence of mutations in the genes encoding NKB ([italic]TAC3[/italic]) and its receptor ([italic]TACR3[/italic]) in GDPP patients.[br][bold]Subjects and Methods:[/bold] Coding and boundary regions of [italic]TAC3 [/italic]and [italic]TACR3[/italic] were sequenced in 96 patients (93F/3M) with GDPP confirmed by basal and/or stimulated LH and normal CNS MRI. Puberty onset was at 5.9[plusmn]2 in girls and 8[plusmn]1 yrs in boys. A family history of GDPP was present in 13.5%. Functional analysis of the variants identified was performed [italic]in vitro[/italic] using HEK-293 cells to test cellular expression, binding affinity, and IP accumulation.[br][bold]Results:[/bold] Two new [italic]TACR3[/italic] variants were identified in patients without a family history of GDPP, but not in 110 healthy controls. A variant c.1321 C[gt]T (p.Arg441Cys) was found in a patient presenting at 6.7 yrs with thelarche, pubarche, and a pubertal LH (1.5 IU/L; IFMA). [italic]In silico[/italic] analysis predicted this variant to be pathological. Compared to WT NK3R, Arg441Cys NK3R exhibited a reduction in cellular expression levels, reduced ligand binding affinity, and impaired NKB-stimulated IP accumulation. A second novel [italic]TAC3R[/italic] variant, c.918 G[gt]A (p.Met306Ile), was present in a patient presenting at 6.8 yrs with thelarche, pubarche, and a pubertal GnRH-stimulatedLH (9.4 IU/L). [italic]In silico [/italic]analysis predicted this variant to be benign which was confirmed [italic]in vitro[/italic]. Cellular expression, ligand binding affinity, and NKB-stimulated IP accumulation and did not differ between Met306Ile and WT NK3R. No variants were found in the [italic]TAC3[/italic] gene.[br][bold]Conclusion: [/bold]We report the first [italic]TACR3[/italic] mutation in a patient with GDPP. Intriguingly, the novel Arg441Cys NK3R mutant significantly impairs receptor signaling. The exact role of NKB in GnRH physiology remains unclear; however, our results broaden the spectrum of human phenotypes associated with loss-of-function [italic]TACR3[/italic] mutations to include GDPP in addition to hypogonadotropic hypogonadism.[br][br]Sources of Research Support: - FAPESP and CNPq - Brazil (SRA). - CAPES - Brazil (ACSR). - Eunice Kennedy Shriver NICHD/NIH through cooperative agreement U54 HD28138 as part of the Specialized Cooperative Centers Program in Reproduction and Infertility Research (UBK) by R01 HD61577 (UBK).[br][br]Nothing to Disclose: ACSR, NDS, KMI, CYT, CEM, MC, RSC, UBK, SRA 2012-06-25T12:15:00 362 2012-06-25T00:00:00 1899-12-30T12:15:00 1519 277 1933 OR30-5 OR29-01 Monday 1617 2012


1614 ENDO12L_OR30-6 ORAL SESSION: Growth [amp] Puberty (11:15 AM-12:45 PM) Characterization of a Large Cohort of Patients with Idiopathic Central Hypogonadism (ICH) Domenico Vladimiro Libri, Elena Guarducci, Elisa Pignatti, Elisabetta Maioli, Fabiana Guizzardi, Roberta Asci, Paolo Duminuco, Csilla Krausz, Manuela Simoni, Mohamad Maghnie, Antonio Agostino Sinisi, Luca Persani, Marco Bonomi Istituto Auxologico Italiano IRCCS, Milano, Italy; University of Florence, Firenze, Italy; University of Modena and Reggio Emilia, Modena, Italy; EO Galliera, Genova, Italy; University of Federico II, Napoli, Italy; 2nd University of Naples, Napoli, Italy ICH is a rare disease, 5-fold more prevalent in males (M) than in females (F), characterized by a complex pathogenesis. It appears frequently in a sporadic form, although several familial cases have been reported in which ICH follows different modes of inheritance. This finding, together with the description of numerous pathogenetic gene variants and the generation of several animal models, justify the idea of a strong genetic component at the basis of ICH. ICH may indeed be associated to several other morphogenetic or inborn defects, such as the osmic defects that identify the Kallmann syndrome (KS). KS includes about 40% of total ICH cases and have been generally considered as a distinct subgroup of ICH. However, the description of several pedigrees including relatives affected either with isolated osmic defects or KS or normoosmic ICH (nICH) justifies the emerging idea of ICH as a complex genetic disease characterized by variable expressivity and penetrance. Here we present the results of a large national ICH series. Among the 271 total index cases, 118 had KS and 153 nICH. The number of familial cases in the two categories was similar (15.2% KS and 16.9% nICH), but the prevalence of associated malformations or cryptorchidism was higher in KS (22% or 35% versus 7.8% or 13.3% in nICH, respectively). Genetic analysis allow us to identify a contributing genetic defects in about 35% of the patients, with a major involvement of KAL-1 (9.2%), FGFR1 (8.9%) and PROKR2 (7.7%) and a rare involvement of variant in the other candidate genes. These variations were absent in 200 control alleles. They were found on a single allele in 87.3% of the cases, whereas biallelic variations affecting either GnRHR or PROKR2 genes were found in 5.4% and digenic defects in the remaining 7.3%. These digenic defects were quite heterogeneous involving elements on different pathways and were equally distributed among KS and nICH patients. In conclusion, although accumulating evidences indicate the existence of common pathogenic mechanisms for KS and nICH, the combination of hypogonadism with associated malformations or cryptorchidism appears more frequent in KS. Systematic analyses of candidate genes indicate that the pathogenesis of both phenotypes is still largely unknown, but a multiple gene involvement may be seen in both cases.[br][br]Sources of Research Support: The financial support of funds for Young Investigators from the Italian Minister of Health (Grant no. GR-2008-1137632) and IRCCS Istituto Auxologico Italiano IRCCS (Ricerca Corrente Funds 05C701) is gratefully acknowledge.[br][br]Nothing to Disclose: DVL, EG, EP, EM, FG, RA, PD, CK, MS, MM, AAS, LP, MB 2012-06-25T12:30:00 362 2012-06-25T00:00:00 1899-12-30T12:30:00 1540 277 1934 OR30-6 OR29-01 Monday 1618 2012


1615 ENDO12L_MON-1 POSTER SESSION: Pregnancy: Uterine-Trophoblast Dialogs (1:30 PM-3:30 PM) Uterine Decidualization Timing and Magnitude Depend Critically on the Coregulator, Repressor of Estrogen Receptor Activity (REA) Yuechao Zhao, Sunghee Park, Milan K Bagchi, Robert N Taylor, Benita S Katzenellenbogen University of Illinois at Urbana-Champaign, Urbana, IL; Emory University School of Medicine, Atlanta, GA Successful implantation and maintenance of pregnancy require the differentiation and transformation of uterine endometrial stromal cells into morphologically and functionally distinct decidualized cells. Although estrogen and progesterone hormones are known to be required for decidualization, the roles of steroid receptor coregulators in the process of decidualization remain largely unknown. To investigate the specific role of the coregulator, Repressor of Estrogen receptor Activity (REA), in the decidualization of human endometrial stromal cells, we first monitored changes in the level of REA during the decidualization process, and we then investigated the impact of reducing the level of REA on the time course and magnitude of decidualization. Human endometrial stromal cells were obtained from normal cycling women at early proliferative phase. The cells were cultured in vitro with exposure to a hormone cocktail containing estradiol, progesterone and 8-bromo-cAMP, and the cells were monitored morphologically and biochemically over a 10 day period. We found that the expression of REA decreased to ca. 50% in stromal cells as differentiation progressed. Consistent with this finding, we observed that stromal cell differentiation was accelerated and strongly enhanced after the reduction of cellular REA by siRNA knockdown, as indicated both by cell morphology analysis and the increased level of prolactin and progesterone receptor, two well-known markers of decidualization. These findings in human endometrial stromal cells were consistent with our observations on decidualization made in mice with heterozygous deletion of REA. An artificial decidualization response model was used to examine the role of REA in stromal differentiation [italic]in vivo. [/italic]Both morphological and gene regulation analyses revealed an enhanced decidualization response in REA heterozygous mice compared with wild type animals. Importantly, the expression of signal transducer and activator of transcription 3 (STAT3), and of the cytokine interleukin-11 (IL-11), essential regulators of decidualization, were enhanced by the decreased level of REA in both human and mouse endometrial stromal cells. Together, these data reveal that REA normally serves as a repressor-like brake on endometrial stromal cell decidualization. The findings highlight that steroid hormone receptors and their coregulators coordinate their actions to provide precise control of the timing and magnitude of uterine decidualization.[br][br]Sources of Research Support: NIH U54 HD055787 as part of the Eunice Kennedy Shriver NICHD/NIH Centers Program in Reproduction and Infertility Research.[br][br]Nothing to Disclose: YZ, SP, MKB, RNT, BSK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 783 291 1951 MON-1 PO31-01 Monday 1619 2012


1616 ENDO12L_MON-2 POSTER SESSION: Pregnancy: Uterine-Trophoblast Dialogs (1:30 PM-3:30 PM) The Role of Bmp Signaling during Embryo Implantation Caterina Clementi, Mark A Edson, Michael J Large, Ruihong Chen, Vesa Kaartinen, John P Lydon, Francsco J DeMayo, Martin M Matzuk Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; University of Michigan, Ann Arbor, MI To achieve a healthy pregnancy, the embryo and the uterus must establish a cross talk that will be essential throughout gestation. The implantation of the embryo takes place in a limited window of time when the uterus is receptive and the embryo has reached the proper stage of development. This crucial synchrony is controlled by the sex hormones through the regulation of the expression of many downstream factors.[br]Several members of the Bone Morphogenetic Protein (BMP) family are expressed in the pregnant uterus of mice. In particular, Bmp2 has been shown to be required to achieve the implantation of the embryo(1). BMPs signal through heterodimers of BMP type 1 and type 2 serine/threonine kinase receptors. A BMP ligand can bind different receptors in different contexts. While the type 2 receptors are essential for recognizing the ligands, the type 1 receptors determine the specificity of the intracellular response. Three type 1 receptors, ALK2, ALK3, and ALK6, are known to mediate BMP signaling, and are all expressed in the pregnant uterus.[br]To study the implantation process, we investigated the effects of deleting each of these receptors in the uterus. Although Alk6 null mice are sterile, this receptor is not required for the decidual reaction that the uterus accomplishes upon embryo attachment(2). We now demonstrate that Alk2 and Alk3 are both essential for mouse fertility. The deletion of Alk3 recapitulates the fertility defects observed in the Bmp2 cko mice; we suggest that Alk3 and Bmp2 are the receptor-ligand pair that regulates the invasion of the embryo into the endometrim and the following uterine decidualization. On the other hand, the deletion of Alk2 does not affect the ability of the embryo to implant; however, without the expression of Alk2 in the uterus, the decidual response initiates but is not maintained and the implanted embryos die at 5.5 dpc. Interestingly, the Alk2 specific ligand Bmp7, is dispensable during decidualization. These data show that the BMP signaling is required to regulate different steps of pregnancy. The characterization of these different models is a valuable tool to dissect the mechanisms that regulate the early pregnancy stages.[br]The importance of these findings can be fully appreciated if we consider that, in the United States, about 10 percent of women (6.1 million) ages 15-44 have difficulty getting pregnant or staying pregnant. In particular, 75% of the lost pregnancies are due to failure at the implantation stage.[br][br](1) Lee KY, Jeong JW, Wang J, Ma L, Martin JF, Tsai SY, Lydon JP, DeMayo FJ. Mol Cell Biol. 2007 Aug;27(15):5468-78. Epub 2007 May 21. (2) Edson MA, Nalam RL, Clementi C, Franco HL, Demayo FJ, Lyons KM, Pangas SA, Matzuk MM. Mol Endocrinol. 2010 Jun;24(6):1251-66. Epub 2010 Apr 2.[br][br]Sources of Research Support: NIH Grant HD32067.[br][br]Nothing to Disclose: CC, MAE, MJL, RC, VK, JPL, FJD, MMM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2297 291 1952 MON-2 PO31-01 Monday 1620 2012


1617 ENDO12L_MON-3 POSTER SESSION: Pregnancy: Uterine-Trophoblast Dialogs (1:30 PM-3:30 PM) CRH-R1 and CRH-R2 Stimulates Estradiol but Inhibits Progesterone Production in Cultured Human Placental Trophoblasts Via Different Signaling Pathways Lu Gao, Xin Ni Second Military Medical University, Shanghai, China Estrogen and progesterone play pivotal roles in the regulation of pregnancy and parturition. Placenta-derived corticotrophin-releasing hormone (CRH), as well as its family members urocortin (UCN) I, II and III are involved in the mechanisms controlling human pregnancy. Previously, we observed that CRH is able to modulate estradiol (E[sub]2[/sub]) and progesterone (P[sub]4[/sub]) production in human placental cells. In the present study, we conducted experiments to determine the roles of CRH receptors, termed CRH-R1 and CRH-R2, as well as their downstream signaling pathways responsible for the regulation of E[sub]2[/sub] and P[sub]4[/sub] production in placenta. We found that either CRH or UCNI antibody dose-dependently reduced E[sub]2 [/sub]while increased P[sub]4 [/sub]concentration in cell culture media. CRH-R1 antagonist antalarmin and CRH-R2 antagonist astressin2b reduced E[sub]2[/sub] but increased P[sub]4[/sub] production, which was consistent with knockdown of CRH-R1 and CRH-R2 using siRNA transfection, respectively. Because of the absence of CRH-R1 specific agonist, CRH-R2 knockdown cells were treated with CRH, and a dose-dependent increase of GTP-bound G[alpha]s protein expression and cAMP concentration was observed, which could be blocked by antalarmin. Application of AC inhibitor SQ22536 and PKA inhibitor H89 blocked CRH-induced E[sub]2[/sub] production in these cells. CRH also stimulated the expression of phosphorylated PLC-[beta]3 (pPLC-[beta]3), which was totally abolished by transfection with a small inhibitory peptide of Gq protein, GP-2A. PLC inhibitor U73122 and PKC inhibitor chelerythrine chloride (CHE) could block CRH-induced E[sub]2[/sub] production and restore P[sub]4[/sub] production. To verify the CRH-R2 coupled signaling pathways, UCNIII, the specific CRH-R2 agonist was added into cell culture. The level of GTP-bound G[alpha]i but not G[alpha]s protein increased, followed by the decrease of cAMP concentration, which was blocked by astressin2b. UCNIII also resulted in an increase of pPLC-[beta]3 expression. The increase of E[sub]2 [/sub]and decrease of P[sub]4[/sub] production caused by UCNIII were both blocked by U73122 and CHE, but not by SQ22536 and H89. In conclusion, activation of G[alpha]s protein by CRH-R1 and its downstream AC-cAMP-PKA signaling predominates the CRH-induced E[sub]2[/sub] production, whereas activation of G[alpha]q protein by both CRH receptors and downstream PLC[beta]3-PKC signaling concurrently contributes to E[sub]2[/sub] increase and P[sub]4[/sub] decrease by CRH treatment. The local change in progesterone/estrogen ratio caused by endogenous CRH and peptides might be important for control of human pregnancy and parturition.[br][br]Sources of Research Support: Natural Science Foundation of China 30900509, 30800391, 81170596.[br][br]Nothing to Disclose: LG, XN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1160 291 1953 MON-3 PO31-01 Monday 1621 2012


1618 ENDO12L_MON-4 POSTER SESSION: Pregnancy: Uterine-Trophoblast Dialogs (1:30 PM-3:30 PM) Corticotropin-Releasing Hormone Modulates Output of IL-6, IL-8 and MCP-1 in Primary Human Myometrial Smooth Muscle Cells Chen Xu, Jie Liu, Xingji You, Stephen Wood, Donna M Slater, David M Olson, Xin Ni Second Military Medical University, Shanghai, China; University of Alberta, Edmonton, Canada; University of Calgary, Calgary, Canada; University of Calgary, Calgary, Canada Corticotropin-releasing hormone (CRH) has been implicated to link to [quot]a placenta clock[rdquo] controling gestational length and triggering the parturition in human. Recent evidence indicates that inflammatory stimulated process may be a key mechanism of human labour. We hypothesized that CRH may induce a cascade of events leading to parturition through the inflammatory signaling in myometrium. The myometrial smooth muscle cells were isolated from lower segment (LS) and upper segment (US) biopsies obtained from pregnant women who were not undergoing labour at term and then cultured. Cultured cells were treated with increasing concentration of CRH for 24h in absence and presence of the specific antagonists of two CRH receptors (CRHR1 and CRHR2). The medium supernatants were then collected for ELISA mass determination for IL-6, IL-8 and MCP-1 output. It was found that CRH treatment increased IL-6, IL-8 and MCP-1 output in both US and LS myometrial cells in a dose-dependent manner. CRH at 10[sup]-6[/sup] mol/L significantly stimulated IL-8 output by 4 fold (P[lt]0.01) in LS while by 2.5 fold (P[lt]0.01) in US. CRH significantly increased IL-6 output by 2-fold (p[lt]0.01) in the LS, but had no significant effect in the US. MCP-1 output in both regions was stimulated 2-3 fold by CRH (10[sup]-6[/sup]M) (p[lt]0.01). In LS, the stimulatory effects of CRH on IL-6, IL-8 and MCP-1 production was blocked by CRHR1 antagonist. Knockdown of CRHR1 by small RNA interference resulted in loss of stimulatory effect of CRH. In US, CRH stimulation of IL-8 production was blocked by CRHR1 antagonist, whereas its stimulation of MCP-1 was reversed by CRHR2 antagonist. These effects were confirmed in CRHR1 knockdown or CRHR2 knockdown cells. For IL-6 production, stimulatory effect of CRH in US was blocked by CRHR1 antagonist. Knockdown of CRHR1 in US cells caused inhibition of IL-6 production upon CRH treatment, suggesting that CRHR1 stimulated whereas CRHR2 inhibited IL-6 output in US. Our results indicate that CRH up-regulate the pro-inflammatory cytokines, such as IL-6, stimulate the production of chemokines, IL-8 and MCP-1, thereby impacting the recruitment of leukocytes to the uterus, resulting in inflammatory reactions and trigger of the uterus for labor and delivery.[br][br]Sources of Research Support: Natural Science Foundation of China (No. 30971090 [amp]30811120433), CIHR [amp] AIHS PreHot in Canada.[br][br]Nothing to Disclose: CX, JL, XY, SW, DMS, DMO, XN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2025 291 1954 MON-4 PO31-01 Monday 1622 2012


1619 ENDO12L_MON-5 POSTER SESSION: Pregnancy: Uterine-Trophoblast Dialogs (1:30 PM-3:30 PM) The microRNA (miR)-17-92 Cluster Regulates Key Placental Genes To Repress Human Trophoblast Differentiation Premlata Kumar, Carole R Mendelson UT Southwestern Medical Center, Dallas, TX Mononuclear cytotrophoblast (CYT) cells of the human placenta proliferate rapidly and subsequently fuse and differentiate to form the multinucleated syncytiotrophoblast (SYN) layer with concomitant induction in aromatase/[italic]hCYP19[/italic] and human chorionic gonadotropin ([italic]hCG[beta][/italic]) expression. To identify differentially expressed miRNAs during human trophoblast differentiation, we performed miRNA microarray analysis of total RNA from CYT and SYN cells. We observed that members of the miRNA-17-92 cluster and its paralog miR-106a-363 were significantly downregulated upon SYN differentiation. Using TargetScan analysis, we identified several predicted mRNA targets of these miRNAs; these included CYP19 mRNA itself, and GCM1, a transcription factor critical for labyrinthine trophoblast development and [italic]CYP19[/italic] expression. Downregulation of the miR-17-92 cluster during SYN differentiation was confirmed using TaqMan-based RT-qPCR. Importantly, overexpression of miR-19b and miR-106a, members of the miR-17-92 cluster, in cultured human trophoblast cells significantly decreased hCYP19 mRNA and protein expression, suggesting hCYP19 mRNA as a target of miR-19b. Overexpression of miR-19b also inhibited expression of GCM1 protein and hCG[beta] mRNA in cultured human trophoblasts. Luciferase reporter assays in JEG3 cells cotransfected with hCYP19- and GCM1-3[apos]UTR-luciferase reporters revealed that hCYP19 and GCM1 mRNAs are direct targets of miR-19b. Mutagenesis of the putative miRNA binding sites in the hCYP19 and GCM1 3[apos]-UTRs confirmed these interactions. GCM1 knockdown in cultured human trophoblasts resulted in decreased [italic]hCYP19[/italic] and [italic]hCG[beta][/italic] expression, substantiating its essential role in trophoblast differentiation. The protooncogene, c-myc, has been reported to induce miR-17-92 expression. Interestingly, c-myc expression dramatically declined upon differentiation of human SYN in culture. Moreover, adenovirus-mediated overexpression of c-myc in cultured human trophoblasts caused upregulation of the miR-17-92 cluster and associated inhibition of [italic]hCYP19[/italic] and [italic]hCG[beta][/italic] expression. Thus, we have established that the c-myc-induced miR-17-92 cluster is highly expressed in undifferentiated, proliferating CYT. Furthermore, these miRNAs target GCM1 and hCYP19 expression and may act to functionally repress human trophoblast differentiation. Conversely, during SYN differentiation, the decline in c-myc causes a decrease in miR-17-92 expression allowing for induction of genes critical for placental differentiation.[br][br]Sources of Research Support: NIH-R01-DK031206.[br][br]Nothing to Disclose: PK, CRM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1786 291 1955 MON-5 PO31-01 Monday 1623 2012


1620 ENDO12L_MON-6 POSTER SESSION: Pregnancy: Uterine-Trophoblast Dialogs (1:30 PM-3:30 PM) Characterization of Gestationally Regulated Estrogen Receptor [alpha] (ESR1) Isoforms in the Pregnant Myometrium Robert C Moore, Ryeung Lee Sang, Christina Kachulis, Arvind Suresh, Evan J Easton, Steve N Caritis, Jennifer C Condon, Pancharatnam Jeyasuria University of Pittsburgh, Pittsburgh, PA Estrogen (E2) signaling through estrogen receptor [alpha] (ER[alpha]) regulates the timing of parturition by increasing the expression of contractile associated proteins (CAPs) as term approaches. ER[alpha] is expressed in multiple tissues during pregnancy, however the high circulating levels of E2 throughout mammalian pregnancy preclude effective signaling at the ligand level. We suggest that the differential expression of two ER[alpha] splice variants (ER[alpha]46 and ER[Delta]7) and the full-length ER[alpha]66 in the pregnant myometrium regulates E2 action in a tissue specific and gestationally regulated manner. We speculate that E2 action in the pregnant mouse and human uterus is limited across gestation despite elevated E2 levels due to the inhibitory action of the alternate ER[alpha] isoforms. At term in both the mouse and human a switch in ER[alpha] isoforms results in the full length ER[alpha]66 isoform becoming the predominant form, thereby promoting an increased estrogenic pro-contractile response. Western analysis of the human myometrium identifies ER[alpha]66, to principally reside in the cytoplasm whereas the truncated ER[alpha]46 (skips Exon1) and the splice variant ER[Delta]7 (skips exon 7) are primarily observed in the nuclear fraction. Myometrial tissue isolated from pregnant women at 30 weeks gestation and term demonstrate a distinct down regulation of the inhibitory isoforms as term approaches. Quantification of the ER[alpha] isoforms through RPA further confirms the decreased levels of ER[Delta]7 at term in the pregnant human myometrium. Similarly in the pregnant mouse uterus the primary isoform observed in the nuclear fraction is ER[alpha]46 isoform, which is replaced at term by ER[alpha]66 isoform. Progesterone treated animals demonstrate a delay in labor associated with a significant decrease in uterine ER[alpha]66 and a prolonged presence of the uterine ER[alpha]46 isoform at E19. siRNA analysis confirmed that in the absence of ER[alpha]66, ER[alpha]46 has the capacity to regulate CAP gene expression. We suggest that alternative splicing via gestationally regulated splicing factors such as SF2/ASF and hnRNPA1 regulate differential uterine ER[alpha] isoform expression across gestation. Specifically we have observed the splice factor SF2 highly expressed in both the pregnant mouse and human uterus with increased expression earlier in gestation. In conclusion we believe that alternative splicing mechanisms lead to alternatively spliced ER[alpha] isoforms, which play an important role in the maintenance of myometrial quiescence.[br][br]Sources of Research Support: HD047905.[br][br]Nothing to Disclose: RCM, RLS, CK, AS, EJE, SNC, JCC, PJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 976 291 1956 MON-6 PO31-01 Monday 1624 2012


1621 ENDO12L_MON-7 POSTER SESSION: Pregnancy: Uterine-Trophoblast Dialogs (1:30 PM-3:30 PM) Caffeine and Its Primary Metabolite Paraxanthine Reduce 11[beta]-Hydroxysteroid Dehydrogenase Type 2 Expression in Human Trophoblast Cells through the Adenosine A[sub]2B[/sub] Receptor Saina Sharmin, Haiyan Guan, Andrew Williams, Kaiping Yang Western University, London, Canada; Western University, London, Canada; Lawson Health Research Institute, London, Canada Maternal caffeine consumption is associated with increased risk of fetal growth restriction (FGR), but the underlying molecular mechanisms are unknown. There is evidence that reduced placental 11[beta]-hydroxysteroid dehydrogenase type 2 (11[beta]-HSD2) is link to FGR, we hypothesized that caffeine induces FGR partly through down regulating placental 11[beta]-HSD2. As a first step in examining this hypothesis, we studied the effects of caffeine on placental 11[beta]-HSD2 activity and expression using our established primary human trophoblast cells as an in vitro model system. Given that maternal serum concentrations of paraxanthine (the primary metabolite of caffeine) were greater in women who gave birth to FGR infants than to appropriately grown infants, we also studied the effects of paraxanthine. Our main findings were: [bold](1)[/bold] both caffeine and paraxanthine decreased placental 11[beta]-HSD2 activity, protein and mRNA in a concentration-dependent manner; [bold](2)[/bold] this inhibitory effect was mediated by the adenosine A[sub]2B[/sub] receptor, since siRNA-mediated knockdown of the expression of this receptor prevented caffeine- and paraxanthine-induced inhibition of placental 11[beta]-HSD2; and [bold](3)[/bold] it is known that adenosine receptors signal through cAMP, we demonstrated that caffeine and paraxanthine inhibition of placental 11[beta]-HSD2 was achieved through decreases in cAMP, because forskolin (an activator of adenyl cyclase) abrogated this inhibitory effect. Taken together, these findings reveal that placental 11[beta]-HSD2 is a novel molecular target through which caffeine and its metabolite may induce FGR. They also uncover a previously unappreciated role for the adenosine A[sub]2B[/sub] receptor signaling in regulating placental 11[beta]-HSD2, and consequently fetal development.[br][br]Sources of Research Support: Canadian Institutes of Health Research.[br][br]Nothing to Disclose: SS, HG, AW, KY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 819 291 1957 MON-7 PO31-01 Monday 1625 2012


1622 ENDO12L_MON-8 POSTER SESSION: Pregnancy: Uterine-Trophoblast Dialogs (1:30 PM-3:30 PM) [italic]Plac1 [/italic](Placenta-Specific 1) Deletion Is Associated with Postnatal Hydrocephalus and Decreased Viability Suzanne Michelle Jackman, Xiaoyuan Kong, Michael Fant University of South Florida, Tampa, FL; University of South Florida, Tampa, FL; University of South Florida, Tampa, FL Background: [italic]Plac1[/italic] is a recently discovered X-linked gene that maps to a region of the X chromosome important in placental and fetal development. We have previously demonstrated that [italic]Plac1[/italic] is paternally imprinted and essential for normal placental development. [italic]Plac1[/italic] mutants inheriting the mutant allele from the mother are associated with placentomegaly and IUGR.[br]Objectives: The aim of this study was to examine the postnatal phenotype associated with [italic]Plac1 [/italic]silencing.[br]Methods: [italic]Plac1[/italic] was deleted in murine ES cells and bred against a C57BL/6 background. Timed matings were established between wild type (WT) or hemizygous males and heterozygous (het) or WT females. Placentae were obtained at various gestational ages and genotyped using a PCR-based strategy. Offspring were genotyped at weaning (21 days) and observed for 4 months, postnatally.[br]Results: The initial analysis of the [italic]Plac1[/italic] mutant mouse was based on a WT male x het female breeding scheme. At weaning, [italic]Plac1[/italic] deletion was associated with fewer than expected KO males (23%) based on analysis of 88 mice. Similarly, when a KO male x het female breeding scheme was used no [italic]Plac1[/italic] homozygous females were identified although the number of mice available from this breeding scheme was limited (n=8). By contrast, when embryos were genotyped prior to birth all genotypes were present at or close to their expected numbers (n=69) suggesting that the observed decrease in viability was due to adverse events that occurred during or shortly after delivery. Interestingly, 10-15% of the X[italic][sup]Plac1[/sup][/italic]X females (mutant maternal allele) and 20% of the surviving X[italic][sup]Plac1[/sup][/italic]Y (KO) males developed lethal hydrocephalus at 4-8 weeks. By contrast, XX[italic][sup]Plac1[/sup][/italic] hets that inherited the mutation from the paternal allele did not exhibit this postnatal phenotype suggesting that it was due to [italic]Plac1[/italic]-dependent events at the placental level. Lastly, [italic]Plac1[/italic] mutant mice that survived and did not develop hydrocephalus exhibited normal postnatal growth.[br]Conclusions: [italic]Plac1[/italic] is a paternally imprinted gene essential for normal placental and embryonic growth. Interestingly, [italic]Plac1[/italic] KO mice exhibit decreased viability postnatally but not prenatally. Additionally, [italic]Plac1[/italic] hets (X[italic][sup]Plac1[/sup][/italic]X) and[italic] Plac1[/italic] KO[apos]s exhibit an increased prevalence of postnatal hydrocephalus whereas hets derived from a mutant paternal allele do not. Collectively, these observations suggest that [italic]Plac1[/italic]-dependent alterations in placental development/function influence CNS development and viability.[br][br]Sources of Research Support: March of Dimes #6-FY09-503.[br][br]Nothing to Disclose: SMJ, XK, MF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2013 291 1958 MON-8 PO31-01 Monday 1626 2012


1623 ENDO12L_MON-9 POSTER SESSION: Pregnancy: Uterine-Trophoblast Dialogs (1:30 PM-3:30 PM) Estrogen-Related Receptor [gamma] (ERR[gamma]) Regulation of Voltage-Gated K[sup]+[/sup] Channels and Kallikrein-Kinin System (KKS) during Human Trophoblast Differentiation Yanmin Luo, Premlata Kumar, Carole R Mendelson University of Texas Southwestern Medical Center, Dallas, TX The placenta is essential for the establishment and maintenance of pregnancy. Reduced oxygenation of the placenta may lead to preeclampsia, a hypertensive disorder that affects 6-8% of all pregnancies in the U.S. and is a leading cause of maternal and neonatal morbidity and mortality. Aromatase/[italic]CYP19[/italic], the critical enzyme in estrogen biosynthesis, is highly expressed in syncytiotrophoblast of human placenta. Previous studies from our lab (1) revealed that ERR[gamma], which is rapidly upregulated during differentiation of human syncytiotrophoblast in culture, serves a critical regulatory role in the associated induction of [italic]CYP19[/italic] gene expression. Both the induction of ERR[gamma] and [italic]CYP19[/italic] are dependent upon a critical O[sub]2[/sub] tension. Recent studies of ERR[gamma] null mice have uncovered unexpected roles in regulation of K[sup]+[/sup] channel subunits, K[sup]+[/sup] homeostasis and genes of the KKS in cardiac, gastric and renal tissues (2). The objective of this study was to investigate the expression of voltage-gated K[sup]+[/sup] channel genes and KKS during differentiation of human trophoblasts in culture and the roles of ERR[gamma] and O[sub]2[/sub] tension in their regulation. To define the mechanisms for trophoblast differentiation in human placenta, we have utilized a model system wherein mononuclear cytotrophoblast cells isolated from mid-gestation human placenta spontaneously fuse and differentiate in culture to form syncytiotrophoblast-like cells. The expression levels of K[sup]+[/sup] channel subunits KCNQ1, KCNE1, KCNE3, KCNE5 and the kallikrein KLK1, analyzed by qRT-PCR, increased during the differentiation of human trophoblast cells in a 20% O[sub]2[/sub] environment. Expression levels of KCNQ1, KCNE3 and KLK1 were markedly reduced when the cells were cultured in a hypoxic environment (2% O[sub]2[/sub]), as was the case for [italic]CYP19[/italic] and ERR[gamma]. Upon transduction of human trophoblast cells with lentiviral vectors expressing shRNAs to knock down ERR[gamma], expression of KCNQ1 was significantly decreased along with the expression of [italic]CYP19[/italic], while expression of the other K[sup]+[/sup] channel genes were unaffected. These findings suggest that ERR[gamma] may mediate O[sub]2[/sub]-dependent expression of genes involved in K[sup]+[/sup] homeostasis during human trophoblast differentiation. Moreover, hypoxia-related defects in ERR[gamma] expression could possibly play a role in the pathogenesis of preeclampsia.[br][br](1) Kumar P, Mendelson CR. Mol Endocrinol 2011; 25:1513-1526. (2) Alaynick WA et al. Mol Endocrinol 2010; 24:299-309.[br][br]Sources of Research Support: NIH-R01-DK31206.[br][br]Nothing to Disclose: YL, PK, CRM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1788 291 1959 MON-9 PO31-01 Monday 1627 2012


1624 ENDO12L_MON-10 POSTER SESSION: Pregnancy: Uterine-Trophoblast Dialogs (1:30 PM-3:30 PM) Transcription Factor FOXF1 Is Critical for Human Trophoblast Differentiation Stuart Handwerger, Cherie Kessler University of Cincinnati, Cincinnati, OH The transcription factors and signaling molecules that are critical for the differentiation of human trophoblast cells are poorly understood. Earlier studies from our laboratory demonstrated that the transcription factor FOXF1 (forkhead box F1, also known as FREAC1), which is important in the regulation of the differentiation of lung and other tissues, is markedly induced during the in vitro differentiation of human mononuclear cytotrophoblast cells (CTB) to a multinucleated syncytiotrophoblast cell (STB) phenotype. Since these studies suggest a possible role for FOXF1 in placenta differentiation, we subsequently examined whether silencing of FOXF1 blocks the syncytialization of CTB and attenuates the expression of STB-specific genes, such as placental lactogen (hPL) and chorionic gonadotropin (hCG). A highly enriched fraction of enzymatically dispersed human CTB that differentiate spontaneously to a STB phenotype was exposed for 3 days to a lentivirus that expresses a FOXF1 shRNA and green fluorescent protein (GFP) or to a control lentivirus that expresses GFP but not the FOXF1 shRNA. Immunohistochemical studies (IHC) with an anti-desmoplakin antibody, which stains cell membranes, were used to assess the extent of syncytialization; and real-time PCR analyses were utilized to assess the effects of FOXF1 silencing on hPL, hCG, human growth hormone-variant (hGH-V) and pregnancy-specific glycoprotein 1 (PSG1) mRNA levels. Preliminary studies indicated that approximately 90 percent of the FOXF1 shRNA-treated and control cells expressed GFP after 3 days of culture, indicating nearly complete uptake of the lentiviruses. Approximately 60 percent of the CTB exposed to the empty lentivirus fused to a multinucleated syncytium with most STB containing 4-5 nuclei. In contrast, only about 20 percent of the CTB exposed to the FOXF1 shRNA formed a syncytium; and most of these STB contained only 2-3 nuclei per cell. In addition, the cells exposed to the shRNA expressed 40-50 percent less of each of the STB marker genes than the cells exposed to the control lentivirus. Taken together, these findings strongly suggest a critical role for FOXF1 in human villous CTB differentiation to a STB phenotype. FOXF1 induces both the fusion of CTB to form a syncytium and the expression of STB-specific genes.[br][br]Sources of Research Support: NIH HD065339.[br][br]Nothing to Disclose: SH, CK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 580 291 1960 MON-10 PO31-01 Monday 1628 2012


1625 ENDO12L_MON-11 POSTER SESSION: Pregnancy: Uterine-Trophoblast Dialogs (1:30 PM-3:30 PM) Increasing Estradiol (E2) Level Increases Cell Death and Decreases Cell Proliferation in an HTR-8/SVneo Cytotrophoblast Cell Line Debra F Skafar, Laura Terlecky, Anthony Imudia, Brian A Kilburn, D Randall Armant Wayne State University School of Medicine, Detroit, MI; Wayne State University School of Medicine, Detroit, MI Inappropriate elevation of E2 levels in the first trimester of pregnancy has deleterious effects on trophoblast function in a baboon model. To determine the underlying mechanism responsible for this counterintuitive observation, growth and survival activities of a human first trimester cytotrophoblast cell line, HTR-8/SVneo, were assessed after culture for 8 or 24 h in the presence or absence of E2. Both ERalpha and ERbeta were expressed in the cell line according to western blotting. Cell death, measured by COMET and TUNEL assay, increased concomitantly with E2 concentration and was attenuated by the E2 antagonist ICI182,780 (p[lt]0.05). The ERbeta-selective ligand, diarylpropionitrile, was without effect, suggesting a mechanism involving ERalpha. Cell proliferation, examined by Ki67 staining, was decreased by E2 (p[lt]0.05). These data suggest that elevated E2 contributes to failed remodeling of the uterine arteries by reducing trophoblast cell number through diminished growth and poor survival.[br][br]Sources of Research Support: The Intramural Research Program of NICHD, by NIH grant HD045966 and by a Research Stimulation Grant from Wayne State University.[br][br]Nothing to Disclose: DFS, LT, AI, BAK, DRA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 774 291 1961 MON-11 PO31-01 Monday 1629 2012


1626 ENDO12L_MON-12 POSTER SESSION: Pregnancy: Uterine-Trophoblast Dialogs (1:30 PM-3:30 PM) The Role of Osteopontin in Early Pregnancy Biserka Mulac Jericevic, Tamara Nikolic, Sandra Sucurovic Medical School, University of Rijeka, Rijeka, Croatia The uterus is a mucosal tissue that undergoes massive steroid-hormone promoted remodeling during pregnancy to support conceptus survival and development. These changes are accompanied by the differentiation and proliferation of a unique, transient uterine natural killer (uNK) cell lineage. In the human, rat and mouse uterus, uNK cells are terminally differentiated cells reaching their highest numbers in mid gestation and are specifically recruited to placental attachment sites where intensive formation of new maternal blood vessels take place. The robust vascular development that occurs during implantation and early placentation is of paramount importance for the development of a receptive endometrium suitable for implantation. It is known that mouse uNK cells express angiogenic molecules, and we found that sialoprotein osteopontin (OPN) is made by granulated (uNK) cells.[br]Osteopontin (OPN) is a secreted extra cellular protein involved in many physiological and pathological processes including vascular remodeling. OPN is expressed during early pregnancy, and OPN null mice manifest decreased pregnancy rates during midgestation when compared with wild-type counterparts. Differentiation of endometrial stromal cells and formation of new maternal blood vessels at the time of embryo implantation are critical processes for the establishment and maintenance of gestation. In non-immune cells OPN signals mainly trough [alpha][sub]v[/sub][beta]3 integrin, and in immune cells trough CD44.[br][alpha][sub]v[/sub][beta]3 has been shown to be highly expressed at the time of embryo attachment, and aberrant expression of [alpha][sub]v[/sub][beta]3 is associated with infertility.[br]We hypothesized that OPN/[alpha][sub]v[/sub][beta]3 signaling mechanisms control of the angiogenic factors important for the vascularization process during early pregnancy. To test this hypothesis mouse and human uterine tissues were used. Our results suggest that that OPN paracrine signaling pathway significantly contributes to angiogenesis during early pregnancy.[br][br]Sources of Research Support: This research is supported by The Croatian Ministry of Science, Education and Sports (to BMJ).[br][br]Nothing to Disclose: BMJ, TN, SS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1205 291 1962 MON-12 PO31-01 Monday 1630 2012


1627 ENDO12L_MON-13 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Dihydrotestosterone Treatment in Mice Induces a Persistent Polycystic Ovary Syndrome Phenotype E Leonie AF van Houten, Piet Kramer, Anke Mcluskey, Bas Karels, Axel PN Themmen, Jenny A Visser Erasmus Medical Center, Rotterdam, Netherlands Polycystic ovary syndrome (PCOS), the most common endocrine disorder in women in their reproductive years, is defined by two out of the following three criteria: hyperandrogenism, oligo/anovulation, and polycystic ovaries. Affected women are often obese and insulin-resistant. In order to study the interaction between ovarian and adipose function we recently developed a mouse PCOS model through chronic exposure to dihydrotestosterone (DHT). Here, we studied whether the PCOS phenotype persists after withdrawal of DHT treatment, which would more closely resemble PCOS in women.[br]Prepubertal female mice were treated with a 90-day continuous release pellet containing DHT or vehicle. After 90 days of treatment, mice were sacrificed (chronically-treated group, n=17), or were allowed a 30-day wash-out period before sacrifice (persistent group, n=17). Vaginal smears were taken to detect estrous cycle abnormalities. Ovaries were collected to determine the presence of cystic follicles. To determine an effect of DHT treatment on metabolism, body weight was measured, an Intraperitoneal Glucose Tolerance Test (IPGTT) (n=8-9 per group) and an Insulin Tolerance Test (ITT) (n=8-9 per group) were performed as a measure for glucose intolerance and insulin resistance.[br]After 90 days, DHT-treated mice were acyclic, their ovaries contained antral follicles with a cyst-like structure and an increased number of atretic follicles, bodyweights were higher, and chronically DHT-treated mice were glucose intolerant.[br]Mice of the persistent group remained anovulatory, their ovaries still contained cyst-like follicles and an increased number of atretic follicles. In contrast to the chronically DHT-treated mice, DHT-treated mice of the persistent group had an increased number of healthy small pre-antral and small antral follicles compared to placebo-treated mice. Bodyweight and weights of fat depots remained significantly increased in the persistent group and mice remained glucose intolerant. Insulin sensitivity did not appear to be affected in the DHT-treated mice, although the counter regulatory mechanisms after insulin administration were impaired in both the chronically DHT-treated and persistent group.[br]In conclusion, we confirmed that chronic DHT-treatment induces a PCOS-like ovarian and metabolic phenotype in mice, which remained even after a 30-day wash-out period. This suggests that in mice after a 90-day DHT treatment period a persistent PCOS phenotype is induced.[br][br]Nothing to Disclose: ELAFvH, PK, AM, BK, APNT, JAV 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1505 292 1963 MON-13 PO31-02 Monday 1631 2012


1628 ENDO12L_MON-14 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Effects of Dehydroepiandrosterone on Female Reproduction in Mice Osamu Suzuki, Minako Koura, Yoko Noguchi, Kozue Uchio-Yamada, Junichiro Matsuda National Institute of Biomedical Innovation, Ibaraki, Japan [Aim] Strain differences in in vitro fertilizability still constitute a serious problem in mouse reproduction. To improve the in vitro fertilizability of mouse oocytes, we examined the effects of implanting time-release pellets of dehydroepiandrosterone (DHEA), a testosterone precursor, in female mice on oocyte fertilizability. [Methods] DHEA pellets (0.25, 1.5, or 5 mg/pellet, 21-day release form, Innovative Research of America) or corresponding placebo pellets were implanted subcutaneously in 9-week-old female 129X1/SvJJmsSlc (129x1) mice. After 21-day treatment, superovulation was induced in these females by hormonal injections. Then, in vitro fertilization was conducted using the oocytes from these females and epididymal sperm from Slc:ICR male mice. Embryos were cultured in vitro for 96 h. Ovarian and body weights, ovulation rates, and in vitro development of the embryos were recorded. [Results and discussion] Neither ovarian nor body weights were significantly different between DHEA and placebo-treated females in any doses. DHEA suppressed the ovulation rates in 1.5 and 5 mg groups (DHEA versus placebo: 21.0 [plusmn] 2.6 versus 32.5 [plusmn] 2.8 and 19.9 [plusmn] 1.4 versus 27.1 [plusmn] 1.6, respectively, n=10, p[lt]0.05), but not in the 0.25 mg group (26.6 [plusmn] 3.2 versus 24.8 [plusmn] 2.4). DHEA did not affect the percentages of 2-cell embryo formation in any doses (50-60 %). In the 0.25 mg group, DHEA treatment tended to increase a blastocyst formation rate (1.8 [plusmn] 0.8% versus 0.4 [plusmn] 0.4%, not significant). But the treatment suppressed the rate in 1.5 mg group (0.0 [plusmn] 0.0% versus 3.1 [plusmn] 0.7%, p[lt]0.05) and did not affect them in 5 mg group (1.3 [plusmn] 0.9% versus 0.8 [plusmn] 0.6%). These results suggests that DHEA at low doses (0.25 mg pellets for 21-days in this study) might have a potential to improve in vitro fertilizability of mouse oocytes. Because high doses of DHEA showed the adverse effect on oocytes, a more detailed study is needed to determine the optimal dose, age, and duration of DHEA treatment.[br][br]Sources of Research Support: This work was supported by a grant from the Ministry of Health, Labour, and Welfare of Japan.[br][br]Nothing to Disclose: OS, MK, YN, KU-Y, JM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1526 292 1964 MON-14 PO31-02 Monday 1632 2012


1629 ENDO12L_MON-15 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Developmental Programming: Gestational Exposure to Excess Testosterone, by Its Androgenic Action, Disrupts Maternal Steroidal and Metabolic Environment in Sheep Bachir A Abi-Salloum, Almudena Veiga-Lopez, David H Abbott, Vasantha Padmanabhan University of Michigan, Ann Arbor, MI; University of Wisconsin, Madison, WI Altered maternal environment reprograms fetal development culminating in postnatal endocrine and metabolic disorders. In sheep, prenatal testosterone (T) treatment leads to low birth weight offspring, postnatal catch-up of growth, as well as adult reproductive and metabolic defects. We tested the hypotheses that the impact of gestational (G) T treatment on altered fetal developmental trajectory involves disruption of maternal steroidal and metabolic milieu and that these disruptions are facilitated via androgenic actions of T and altered maternal insulin homeostasis. The study included four treatment groups: control (C; n=12), gestational T (GT, twice weekly i.m. injections of 100 mg of T propionate from day 30 to 90 of gestation; n=12), GT plus an androgen antagonist, flutamide, 15 mg/kg/day orally (GTF; n=10), GT plus an insulin sensitizer, Rosiglitazone, 0.11 mg/kg/day orally (GTR; n=10). Weekly blood samples were collected for assessment of insulin, glucose, maternal steroids (all animals for progesterone [P] and insulin, and a subset [n=6/group], on days 30, 51, 72, and 90 of gestation, for other steroidal measures by liquid chromatography[ndash]mass spectrometry). In addition, fasting insulin and glucose levels were determined on day 90 of gestation (end of treatment). Repeated measures analyses (Mixed model) was performed to test time and treatment effects and their interaction, with weight of ewes and number of fetuses adjusted as covariates, and Bonferroni correction made for multiple comparisons. GT treatment increased (P[lt]0.05) maternal insulin/glucose ratio, T, and androstenedione (A) and decreased P from days 30 to 90 of gestation with no effects on cortisol, estradiol, estrone, and pregnenolone. At gestational day 90 (end of treatment), maternal levels of T (12.0[plusmn]2.4 ng/ml), A (180.0[plusmn]19.3 pg/ml), P (16.9[plusmn]1.3 ng/ml), and insulin (15.6[plusmn]1.8 [micro]U/ml) were higher (P[lt]0.05) in the GT group relative to controls (T: detection limit of assay, A: 100.0[plusmn]0.0, P: 26.6[plusmn]2.8, insulin: 10.6[plusmn]0.6). Co-treatment with flutamide negated the effects of GT on insulin (9.9[plusmn]0.7), and P (22.3[plusmn]2.4). Co-treatment with insulin sensitizer also negated effects of GT on insulin (11.8[plusmn]1.3) as well as A (140.0[plusmn]8.0) but not P (15.3[plusmn]1.6). These findings suggest that impact of GT on fetal developmental trajectory is facilitated by androgenic actions of T via disrupted maternal insulin homeostasis and reduced progestogenic support to the fetus.[br][br]Sources of Research Support: NIH Grant PO1 HD44232 awarded to VP.[br][br]Nothing to Disclose: BAA-S, AV-L, DHA, VP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1365 292 1965 MON-15 PO31-02 Monday 1633 2012


1630 ENDO12L_MON-16 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Naturally Occurring Hyperandrogenism and Intermittent Menstrual Cycles in Female Rhesus Monkeys David H Abbott, Daniel A Dumesic, Kenneth C Lewis, Amber K Edwards, Kim Wallen, Mark E Wilson, Jan D Wagner, Susan E Appt University of Wisconsin, Madison, WI; University of Wisconsin, Madison, WI; University of California, Los Angeles, CA; OpAns LLC, Durham, NC; Yerkes National Primate Research Center, Atlanta, GA; Emory University, Atlanta, GA; Wake Forest School of Medicine, Winston-Salem, NC An epigenetic mimic of polycystic ovary syndrome (PCOS) is induced by [italic]in utero[/italic] testosterone (T) exposure in female macaque monkeys (1). Progress in treating women with PCOS has been limited by incomplete knowledge of PCOS pathogenesis and absence of naturally occurring PCOS in animal models. A female macaque monkey, however, with naturally occurring hyperandrogenism, anovulation and polyfollicular ovaries, accompanied by insulin resistance and increased adiposity (2), suggests non-experimental origins for PCOS in nonhuman primates. As part of a larger study, circulating serum concentrations of T, androstenedione (A) and estradiol (E) were determined by LC/MS/MS from a single, morning blood sample obtained from 120 healthy, ovary-intact, adult female rhesus monkeys ([italic]Macaca mulatta[/italic]) that were not pregnant or nursing an infant, and were under sedation for somatometric measurements. Computerized records provided menstrual histories. The monkeys were of prime reproductive age (8.3[plusmn]0.3 years, mean[plusmn]SEM) and represented a typical spectrum of body weight (7.4[plusmn]0.2 kg; max. 12.5, min. 4.6 kg). Females were identified as having normal (n=99) or high T levels (n=21; [gt]1SD of the overall mean, 0.31 ng/ml, closely resembling an earlier rhesus monkey criterion (1)). In the high T group alone, anogenital distance, a measure of fetal testosterone exposure (3), positively correlated (p[lt]0.015, r[sup2]=0.27) with combined androgen (T+A) levels, suggesting that high adult androgen levels may reflect modest [italic]in utero[/italic] T exposure in female rhesus monkeys. While oligomenorrhea was not pronounced in the high T group, four ([sim]19%) females in this group exhibited intermittent menstrual cycles ([gt]34 days (4)), being otherwise unremarkable, compared to 10 ([sim]10%) females in the normal T group exhibiting intermittent cycles. Taken together, our initial results suggest [sim]3% incidence of hyperandrogenism with intermittent menstrual cycles in adult female rhesus monkeys, without accompanying high BMI. Ongoing studies in cynomolgus ([italic]M. fascicularis[/italic]) and an additional rhesus monkey population will further clarify naturally occurring PCOS-like traits in female macaque monkeys.[br][br](1) Xu N et al., PLoS ONE 2011; 6:e27286. (2) Arifin E et al., Vet Pathol. 2008; 45:512. (3) Herman RA et al., Horm Behav. 2000; 38:52. (4) Abbott DH et al., Am J Primatol 2009; 71:776.[br][br]Sources of Research Support: NIH grants P51 RR000167-48S1 (DHA), P51 RR000165 (YNPRC) and P40 RR021380 (JDW).[br][br]Disclosures: DHA: Ad Hoc Consultant, Viamet Pharmaceuticals, Inc.; Recipient Award, Viamet Pharmaceuticals, Inc. KCL: Chief Executive Officer, OpAns, LLC; Recipient Award, Viamet Pharmaceuticals, Inc. AKE: Researcher, Viamet Pharmaceuticals, Inc. Nothing to Disclose: DAD, KW, MEW, JDW, SEA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1927 292 1966 MON-16 PO31-02 Monday 1634 2012


1631 ENDO12L_MON-17 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) BMI-Increasing Alleles in PCOS Susceptibility Yvonne V Louwers, Blanca M Herrera, Lisette Stolk, Stephen Franks, Andre G Uitterlinden, Joop SE Laven, Mark I McCarthy Erasmus Medical Center, University Medical Center, Rotterdam, Netherlands; University of Oxford, Oxford, UK; Erasmus Medical Center, University Medical Center, Rotterdam, Netherlands; Imperial College London, Hammersmith Hospital, London, UK; Churchill Hospital, Oxford, UK [bold]Introduction [/bold]The majority of women with Polycystic Ovary Syndrome (PCOS) suffer from overweight or obesity, for which several genetic variants have been identified by genome wide association studies (GWAS). Aim of this study was to determine to which extent these variants contribute to risk of PCOS. Since individual effects are modest, we also evaluated the combined effect of these variants on PCOS susceptibility using a genetic risk score calculation.[br][bold]Methods[/bold] Polymorphisms were selected in loci (indicated by closest or most likely candidate gene) which were genome wide significantly associated with Body Mass Index (BMI): BDNF; FAIM2; ETV5; FTO; GNPDA2; KCTD15; MC4R; MTCH2; NEGR1; SEC16B; SH2B1; and TMEM18. PCOS subjects were recruited from the United Kingdom (636 cases and 799 controls) and The Netherlands (523 cases and 2695 controls). PCOS was diagnosed according to the 2003 Rotterdam criteria. The UK cases had an average BMI of 26.0 kg/m[sup]2[/sup] and the controls ([plusmn] 10.9 SD) had a BMI of 25.94 kg/m[sup]2[/sup] ([plusmn] 4.57 SD), whereas the Dutch cases had a average BMI of 27.44 ([plusmn] 6.4 SD) and the controls had a BMI of 26.75 kg/m[sup]2[/sup] ([plusmn] 4.07 SD). Association analysis was carried out within each cohort using the additive genetic model followed by a fixed-effects meta-analysis. The genetic risk score was calculated to determine the risk of developing PCOS for individuals carrying several BMI-increasing alleles.[br][bold]Results [/bold] The SH2B1 variant was significantly associated with PCOS in the Dutch cohort (OR=1.28, 95%CI 1.12-1.47, p=0.0001), but in the meta-analysis did not reach the threshold for significance. None of the other variants, including FTO, showed association with PCOS. The 3% higher BMI of the Dutch controls compared to BMI of the controls from the United Kingdom could influence these results. Genetic Risk Score calculation (GRS) in the total dataset demonstrated that individuals carrying more than 18 BMI-increasing alleles (representing 2% of the population) had a 2.0-fold increased risk of developing PCOS (95%CI=1.1-3.5, p= 0.02).[br][bold]Conclusion[/bold] If we account for BMI, limited additional impact for BMI-increasing alleles on PCOS risk exists. Although the combined presence of more than 18 BMI-increasing alleles shows promising results towards an increased risk of developing PCOS, larger replication efforts are needed to explore this finding further.[br][br]Nothing to Disclose: YVL, BMH, LS, SF, AGU, JSEL, MIM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1921 292 1967 MON-17 PO31-02 Monday 1635 2012


1632 ENDO12L_MON-18 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Anti-Mullerian Hormone May Contribute to Anovulation in PCOS Via Inhibition of Human Granulosa Cell Growth Nafi Mehmet Dilaver, Delecia Goes, Suman Rice, Helen Diane Mason St Georges, University of London, London, UK; St Georges, University of London, London, UK Anti-Mullerian hormone (AMH) is produced by fetal Sertoli cells and causes regression of the Mullerian ducts by a wave of apoptosis during sexual differentiation. AMH and its receptor have now also been localised to granulosa cells (GC) in the ovary postnatally. The precise role of AMH within the ovary is unknown, but it appears to inhibit follicle progression by reducing FSH receptor and aromatase expression and for this reason it is absent in dominant follicles. During folliculogenesis the highest expression is in small healthy antral follicles: the group from which the dominant follicle will be selected. We hypothesised therefore that AMH could not be having a similar apoptotic effect on the granulosa cells within these follicles, to that seen in the Mullerian duct, assuming the same receptor and signalling system is present. Interestingly, although AMH production by GC from small follicles from normal ovaries was [lt]2 ng/ml, from ovulatory polycystic ovaries (PCO) it was 7 ng/ml and from anovulatory PCO, 20 to 40 ng/ml. This suggests a possible role in inhibition of folliculogenesis in polycystic ovary syndrome (PCOS) and we further hypothesised that an inhibitory effect of AMH on cell growth may also be exerted at these higher concentrations. In order to test the effect of AMH on GC growth, KGN cells (granulosa cell line) were cultured with a range of AMH doses from 0.5 to 50ng/ml over eight days. Medium was replenished on days 2, 4 and 6 and cell number was measured on each consecutive day using the MTT assay. The presence of AMHRII and the SMAD signalling system in KGN cells was demonstrated. AMH had no effect on cell growth between 0.5 and 5ng/ml, but was inhibitory of cell growth at days 6, 7 and 8 at 10ng/ml, with further inhibition at 20 and 50ng/ml (ANOVA, p [lt] 0.005). This indicates that at levels produced in normal follicles, AMH would not be inhibitory of cell growth, whereas in anovulatory PCOs the intrafollicular levels could prevent GC proliferation further contributing to anovulation.[br][br]Sources of Research Support: Thomas Addison Scholarship.[br][br]Nothing to Disclose: NMD, DG, SR, HDM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 937 292 1968 MON-18 PO31-02 Monday 1636 2012


1633 ENDO12L_MON-19 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Action of Vitamin D in the Human Ovary: A Novel Effect on Aromatase Suman Rice, Joanna Rachel Anderson, Christiana Georgiou, Henrietta Philippa Brain, Gul Bano, Helen Diane Mason St George[apos]s University of London, London, UK; St George[apos]s University of London, London, UK; Stoke Mandeville Hospital, Aylesbury, UK; St George[apos]s Hospital, London, UK There is increasing awareness of the essential role of vitamin D in many areas of normal physiological functioning. Interestingly, studies have also pointed to a fundamental role of vitamin D (VD) in reproductive function (eg vitamin D-deficient female rats have reduced fertility not correctable by calcium supplementation)(1). We were the first to demonstrate the presence of VD receptor (VDR) in the human ovary by immunohistochemistry, which led us to investigate VDs effects on human granulosa cells (GC) in culture. To our surprise 1,25-dihydroxyvitamin D3 (VD3) inhibited basal and LH-stimulated oestradiol production by GCs dose-dependently, but only once cells had acquired LH receptors, leading us to speculate that VD has a role in ovulation. Women with PCOS are more likely to suffer from VD deficiency and one trial demonstrated improved ovulation frequency with vitamin D supplementation. We hypothesised that VD insufficiency contributes to the aetiology of anovulatory PCOS in these women by affecting aromatase expression. We first determined that the granulosa cell line used (KGN) expressed VDR using standard PCR. Cells were then cultured with forskolin (to mimic the effect of LH in stimulating cAMP)[plusmn]VD3 at 0.02, 0.2, 2 and 20nM, with testosterone (5x10-7M) as an aromatase substrate. After 48h, reverse transcribed RNA was quantified for aromatase mRNA using real-time PCR and normalised against L19. To investigate aromatase promoter II (PII) activity, cells were transfected with a PII-specific reporter construct as well as Renilla expression vector as a transfection control. After serum-starvation, cells were treated as described above and luciferase reporter assays carried out. As expected, forskolin up-regulated aromatase mRNA expression 50-fold and interestingly VD3 reduced this stimulation by half but only at the lowest two doses used with the highest two doses having no effect. This was a direct effect on PII as seen by the reduction in forskolin-stimulated PII activity by VD3 at 0.02 [amp] 0.2nM but not 2 [amp] 20nM. Serum levels less than 50nM (20ng/ml) of 25-hydroxy VD indicate deficiency, with severe deficiency at [lt]12.5nM (2). Hence our in vitro data showed that low levels of VD3 (correlating to serum levels indicative of VD deficiency) substantially reduced the cAMP-driven aromatase expression, which in theory could result in impaired folliculogenesis/ovulation in women with PCOS.[br][br](1)Kwiecinski GG, Petrie GI, De Luca HF [apos]1,25-Dihyroxyvitamin D3 restores fertility in vitamin-D deficient female rats[apos] (1989) Am.J.Physiol. 256:E483-487. (2)The Endocrine Society Clinical Guidelines (2011) [apos]Evaluation, Treatment and Prevention of Vitamin D deficiency[apos].[br][br]Sources of Research Support: St George[apos]s University of London.[br][br]Nothing to Disclose: SR, JRA, CG, HPB, GB, HDM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 928 292 1969 MON-19 PO31-02 Monday 1637 2012


1634 ENDO12L_MON-20 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Adrenocortical Steroid Response to ACTH in Different Phenotypes of Non-Obese Polycystic Ovary Syndrome Nese Cinar, Ayla Harmanci, Duygu Yazgan Aksoy, Kadriye Aydin, Bulent Okan Yildiz Hacettepe University School of Medicine, Ankara, Turkey [bold]Background and aim: [/bold]Adrenal androgen excess is frequently observed in PCOS. We aimed to determine whether adrenocortical steroid response to ACTH varies among patients with different phenotypes of PCOS according to Rotterdam criteria, women with hyperandrogenism only and healthy women.[br][bold]Methods: [/bold]A total of 143 non-obese patients with PCOS (age: 22.2[plusmn]4.1y, BMI:22.5[plusmn]3.1kg/m[sup]2[/sup]), 24 women with hyperandrogenism only and 39 control women were recruited. The groups were matched for age and BMI. Hyperandrogenism (H) was defined as having hirsutism (mF-G score [ge] 6) and/or free androgen index (FAI) [gt] 95% percentile of healthy population. Among women with PCOS, 50 had H, oligo-anovulation (O), and polycystic ovaries (P) (HOP), 32 had H and O (HO), 23 had H and P (HP), and 14 had O and P (OP). Control group was comprised of healthy women without H, O or P. At baseline, total testosterone (T), SHBG, and DHEAS levels were measured. All participants underwent ACTH stimulation test and serum 17-hydroxprogesterone (17-HP), androstenedione (A4), DHEA and cortisol levels were determined at 0, 30 and 60 min. Areas under the curve (AUC) for 17-HP, A4, DHEA and cortisol responses were calculated.[br][bold]Results: [/bold]T, FAI and DHEAS, and basal and AUC values for 17-HP and A4 were significantly and similarly higher in PCOS and H groups than controls (p[lt]0.05 for all) whereas three groups did not differ for basal or AUC values of DHEA and cortisol. Within the PCOS group, 3 hyperandrogenic subphenotypes (HOP, HO, and HP) compared to non-hyperandrogenic subphenotype (OP) had significantly and similarly higher T, FAI, DHEAS and AUC values for 17-HP and A4 (p[lt]0.05). All subphenotypes had similar basal and AUC values for DHEA and cortisol. [br][bold]Conclusion: [/bold]Non-obese patients with PCOS and women with H only have similar adrenal responses that differ from age- and BMI-matched healthy women with higher basal DHEAS, and higher 17-HP and A4 levels both basally and in response to ACTH. Even though H was defined by the presence of hirsutism and/or increased FAI in this study, all three hyperandrogenic subphenotypes exhibited similar adrenal androgen secretion patterns with higher DHEAS, and higher 17-HP and A4 responses to ACTH stimulation compared to non-hyperandrogenic subphenotype. Similar DHEA levels basally and in response to ACTH in hyperandrogenic, non-hyperandrogenic and control women despite higher DHEAS levels might suggest differences in expression of DHEA-sulfotransferase activity.[br][br]Nothing to Disclose: NC, AH, DYA, KA, BOY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1478 292 1970 MON-20 PO31-02 Monday 1638 2012


1635 ENDO12L_MON-21 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Effects of Genetic Background on Insulin Resistance and Fertility in a Polycystic Ovarian Syndrome Mouse Model Abigail Rebecca Dowling, Laura Beth Nedorezov, Joseph Scott Marino, XiaoLiang Qiu, Jennifer Wooten Hill University of Toledo, College of Medicine, Toledo, OH; University of Toledo, Toledo, OH Polycystic ovarian syndrome (PCOS) is the most common endocrine disorder in reproductive age women, and is characterized by hyperandrogenemia, insulin resistance, and reproductive abnormalities. Which of these factors triggers the pathogenesis of PCOS remains unclear. A mouse model of PCOS has been developed using chronic administration of dehydroepiandrosterone (DHEA) to a prepubertal female in the BALB/cByJ strain (1). We hypothesized that the PCOS phenotype would be more pronounced on the diabetes-prone C57/Bl6 background. C57Bl6 and BALB/cByJ female mice at 25 days of age were injected daily with DHEA (6mg/kg body weight) or vehicle for 20 consecutive days. In order to determine which strain more closely mimics PCOS; body weight, body composition, food intake, fasting insulin, glucose tolerance, serum estradiol, testosterone, and insulin were all compared to the control groups of each strain. Hyperandrogenemia induced glucose intolerance in the C57Bl6 but not the BALB/cByJ strain. The fasting insulin levels were dramatically different between the strains of mice, but not between the treated and control mice. Both strains exhibited accelerated puberty and irregular cycles. Analysis of gene and protein expression was then performed for genes known to be linked to PCOS and/or that differ between the strains, including HSD17[beta], POMC, ACVR2A, SGTA, Progesterone receptor, insulin receptor, Cyp17, Cyp11a, StAR, Bcl2, C4b, Gpd1, LHreceptor, Ncam1, Mbp, and Adh1. Among the differences seen, the expression of Fem1B, a gene associated with PCOS in humans, was upregulated in whole ovary of the C57BL6 strain, but not the BALB/c strain. This gene plays a role both in sex determination and glucose tolerance in mice, and thus provides a potential pathway for the induction of insulin resistance in PCOS. Nevertheless, the similarity of the reproductive phenotype in these two strains suggests that susceptibility to glucose dyregulation per se is not a causal factor in the development of PCOS in this model.[br][br]Luchetti, CG et al., Journal of Reproductive Immunology 2004; 64: 59.[br][br]Nothing to Disclose: ARD, LBN, JSM, XQ, JWH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1542 292 1971 MON-21 PO31-02 Monday 1639 2012


1636 ENDO12L_MON-22 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Visfatin Increases IGF-1-Induced Progesterone and Estradiol Secretion in Human Granulosa Cells Maxime Reverchon, Marion Cornuau, Christelle Rame, Dominique Royere, Fabrice Guerif, Joelle Dupont Institut National de la Recherche Agronomique, Nouzilly, France; Centre Hospitalier Universitaire, Tours, France Visfatin, known as a pre-B cell colony-enhancing factor or Nicotinamide phosphoribosyltransferase (Nampt) is a 52-kDa protein recently identified as an adipokine. It is predominantly expressed in and secreted from visceral adipose tissue, exerting a number of insulin mimetic and antagonistic effects. Plasma visfatin concentrations are elevated in the majority of cases of human obesity but also in insulin resistance states, type 2 diabetes mellitus, and polycystic ovary syndrome (PCOS). To date, no role of this adipokine in reproductive functions has been clearly described yet. In the present study, we identified visfatin in human ovarian follicles. We also investigated the effects of human recombinant visfatin on steroid production and on the activation of various signaling pathways in granulosa cells from women undergoing in vitro fertilization and in KGN cells.[br]By RT-PCR and immunoblotting we showed that visfatin is expressed in human granulosa cells and less abundantly in theca cells. By immunohistochemistry we confirmed these results and found also visfatin in human oocyte and cumulus cells. Recombinant human visfatin (10 ng/ml) significantly increased IGF-1 (10-8M)-induced progesterone and estradiol secretions as determined by radioimmunoassay. In contrast, it did not affect steroid secretion in the absence or in the presence of FSH (10-8M) in primary human granulosa and KGN cells. In these cells, recombinant human visfatin also increased IGF-1-induced thymidine incorporation and activated rapidly MAPK ERK1/2, MAPK P38 and Akt phosphorylation but not AMPK phosphorylation under basal conditions (no IGF-1 or FSH). We are now investigating whether visfatin expression is regulated by various insulin sensitizers in human granulosa cells.[br]Taken together, these results confirm the presence and biological activity of visfatin in human granulosa cells. It increases IGF-1-induced steroidogenesis and cell proliferation in primary human granulosa cells and KGN cells. Moreover, it also able to activate several signaling pathways such as MAPK ERK1/2 and p38 and Akt in primary human granulosa cells.[br][br]Nothing to Disclose: MR, MC, CR, DR, FG, JD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 499 292 1972 MON-22 PO31-02 Monday 1640 2012


1637 ENDO12L_MON-23 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Activating G-Protein [alpha]-Subunit (GNAS) Mutation (R201C) in a Virilizing Ovarian Leydig Cell Tumor That Responded to GnRH-Analogue Administration Beatriz Lecumberri, Laura Yebenes, Intza Garin, Jesus Solera, Karen Heath, David Hardisson, Guiomar Perez de Nanclares, Luis Felipe Pallardo La Paz University Hospital, Madrid, Spain; La Paz University Hospital, Madrid, Spain; Txagorritxu Hospital, Vitoria, Spain; La Paz University Hospital, Madrid, Spain Amino-acid substitution at codon 201 or 227 in G-protein alpha-subunit (GNAS) leads to permanent activation of Gs[alpha] subunit and accelerated cAMP production in the absence of stimulatory hormone. These activating mutations are responsible for the McCune-Albright Syndrome and have also been identified in sporadic pituitary and thyroid adenomas, pancreatic cystic neoplasms, adrenocortical lesions, liver, colorectal, renal, and thyroid tumors, and in myelodysplastic syndromes. However, up to this date, there are only 3 reports of GNAS-activating mutations (R201C) in ovarian Leydig Cell Tumors (LCT) (1). All of them were younger than 53, and only in the youngest a response to GnRH-analogue could be demonstrated. LCT are rare gonadal neoplasms that account for less than 0.2% of all ovarian tumors and lead to hyperandrogenism in about 50% of patients. We report a 72-year-old woman with a virilizing LCT harbouring an activating-GNAS mutation (R201C) that responded to LHRH administration. She consulted in 2003 because of a 1-year history of progressively worsening signs of virilization. Extremely high serum testosterone levels were confirmed (12.038 and 30.000 pg/ml, NR: 0,2-2 pg/ml) together with suppressed gonadotropin levels. Ultrasound revealed a 10-mm tumor in the left ovary. Histopathological study after total hysterectomy and bilateral oophorectomy performed in April 2003, described a well-encapsulated left 10 mm LCT, with several plump rod-shaped intracytoplasmic crystals of Reinke[apos]s crystals. Tumor cells showed intense and diffuse cytoplasmic immunoreactivity for [alpha]-inhibin. Postoperatively testosterone levels instantly dropped into the normal range, and virilization slowly regressed. Nine years later the patient remains asymptomatic and cured. She is also a heterozygous carrier of the hemochromatosis H63D mutation and shows a nodular enlargement of her left adrenal gland and a deletion in CYP21A1P pseudogene. Our findings suggest that the activating-GNAS R201C mutation and subsequent cAMP increase may play a significant role in the pathogenesis of virilizing LCT through the stimulation of both gonadal steroid production and tumor development, retaining their responsiveness to gonadotropins. Testosterone levels seem to correlate with age. An active search for GNAS R201C mutation in all LCT might reveal new useful genotype-phenotype correlations. We propose to include virilizing LCT in the widening spectrum of GNAS-related tumors.[br][br](1) Fragoso MC et al., J Clin Endocrinol Metab 1998;83:2074-2078.[br][br]Nothing to Disclose: BL, LY, IG, JS, KH, DH, GPdN, LFP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 956 292 1973 MON-23 PO31-02 Monday 1641 2012


1638 ENDO12L_MON-24 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Prenatal Androgen Exposure Imprinting Establishes Gender Identity Mohammed NMN Ahmed, Abdulraof NMN Almahfouz, Abdulrahman NMN Al Nuaim King Faisal Specialist Hospital [amp] Research Center, Riyadh, Saudi Arabia Background:[br]The underpinning of gender identity is a complex interplay of genetic, hormonal, neuronal [amp] social expectations. It has been difficult to dissect the importance of genetic versus hormonal impact on gender identity.Controversy prevails whether prenatal androgen exposure predict gender identity or not.[br]Aim:[br]We report a case of Congenital Adrenal Hyperplasia with 46, XX karyotype presenting in adult life w/complete virilization [amp] established male identity. Findings here indicate gender identity may be engraved in brain early in life, regardless of genetic compliment [amp] how puberty unfolds.[br]Case Summary:[br]An 18-yr old person w/ambiguous genetalia was initially raised as a female. However, at pubertal age amenorrhea, lack of breast development, excess body hair, were noted. By this time there was well established male sexual identity w/necessary legal work completed by pt. [amp] family. Pt was lost to FU till age 27 when person presented to Urology for reconstructive genital surgery [amp] was referred to Endocrine. O/E: Phenotypic male, muscular, fully breasted beard, mustache,[amp] thick body hair, short statured (ht: 145 cm),worked as male clerk.Pt. had ambiguous external genetalia, prominent labial folds, 5 cm clitoromagaly, w/redundant foreskin, [amp] corona, [amp] urogenital sinus w/5 cm long vagina. Genitogram: cervix, hypoplastic uterus, tubes [amp] vagina identified.US/CT Pelvis: hypoplastic uterus, thin endometrium, Cervix, rudimentary ovaries [amp] tubes. CT Abdomen:Symmetrically enlarged adrenal glands. Karyotype: 46, XX. Hormonal Evaluation: Baseline values 17-OH-Progesterone: 538nmol/l(nl [lt]6.7), 17-OH- Prgenenalone: 278 ng/dl (nl, 53-357), cortisol 202nmol/l(RR:170-536), Post Synacthen stimulation values 649/4550/371 respectively. Serum ACTH 138 ng/l (nl, 5-60). Serum testosterone 20.19 nmol/l (nl, male 9.9-27.8), DHEAS 18.65 umol/l (nl, 4.2-11.5), E2 149 pmol/l(nl, 46-774),FSH 0.8/LH [lt]0.1 IU/l. Pt. insisted on corrective surgery. Mgmt plan: Psychological Support, Surgery recommended augmentation phaloplasty, hysterectomy, vaginoplasty w/UG sinus to continue as voiding source.[br]Conclusions:[br]Prenatal androgen exposure in a genetic female predicts male gender identity. Pts. w/CAH [amp] complete virilization have high risk of being diagnosed late in life. For 46, XX pts. w/established male gender identity it is appropriate to augment male gender identity, undertake hysterectomy/oopherectomy, as has been practiced in different cultures in Middle East [amp] West.[br][br]Nothing to Disclose: MNMNA, ANMNA, ANMNAN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 449 292 1974 MON-24 PO31-02 Monday 1642 2012


1639 ENDO12L_MON-25 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) The Manifestation of Polycystic Ovary Syndrome after the Use of Isotretinoin for Treatment of Severe Acne in a Young Female Pooja Raghavan, Genevieve DeKiel, Charles Katz Mount Carmel Health, Columbus, OH; Mount Carmel Health, Columbus, OH Isotretinoin is commonly used in the treatment of severe cystic acne, and acne which is refractory to other treatment modalities. Although infrequent, metabolic disturbances such as hypertriglyceridemia and elevations in total cholesterol levels have been reported side effects of this medication. Side effects such as these may contribute to the development of insulin resistance in some patients.[br]A 14-year old female with no significant past medical history presented to an outpatient clinic for treatment of severe diffuse cystic acne that was not responsive to treatment with topical medications or oral antibiotics. The patient had normal vital signs including blood pressure measurements, and body mass index was measured as 23.2 kg/m2. Physical exam revealed severe cystic acne over the entire face and upper chest. Laboratory studies, including chemistry panel and liver function tests, were within the normal range. The patient reported starting menses at age 12, and menstrual periods had been irregular since onset. Isotretinoin was prescribed for a 12-week course initially. Six months after the completion of this treatment course, therapy with isotretinoin was repeated for a total of 20 weeks. The patient reported a slight improvement in acne at the end of two treatment courses with isotretinoin. She also reported coarse hair growth underneath the chin and on the lower abdomen. Continued menstrual irregularities and hirsutism until the age of 21 prompted an appointment with a physician. Biochemical studies were checked at this visit, which revealed an elevated LH to FSH ratio, elevated testosterone levels, and glucose intolerance. A transvaginal ultrasound revealed the appearance of polycystic ovaries. The diagnosis of PCOS was made according to the Rotterdam criteria.[br]Patients with PCOS have been reported to have similar lipid abnormalities to those seen in patients being treated with isotretinoin. Insulin resistance has been implicated in the pathogenesis of PCOS. It has been suggested that a combination of both genetic and environmental factors lead to manifestation of PCOS. Thus, clinicians should be aware of potentials agents that could cause this to occur. While isotretinoin is used for the treatment of severe acne in female patients, it may be causing metabolic derangements that worsen the biochemical abnormalities seen in PCOS. This may be causing the syndrome to manifest at an earlier age in patients with a genetic predisposition to PCOS.[br][br]Nothing to Disclose: PR, GD, CK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1687 292 1975 MON-25 PO31-02 Monday 1643 2012


1640 ENDO12L_MON-26 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) The Effects of a Resveratrol Derivative on Regulatory Behaviors and Reproductive Health in Female Long-Evans Rats Kimberly Fabick, Crystal Blake, Merritt Andrus, Edwin Lephart Brigham Young University, Provo, UT; Brigham Young University, Provo, UT; Brigham Young University, Provo, UT Phytoestrogens are naturally occurring compounds found in plants that are structurally similar to estrogens and can bind to mammalian estrogen receptors. Resveratrol is the most studied polyphenolic molecule with positive board health implications [that is associated with the French Paradox (i.e. low incidence of heart-related disease in spite of consuming a high calorie/high fat diet)]. In this study, we examined the resveratrol derivative, 4-acetoxy resveratrol (4AR). The purpose of this study was to determine, in any, side effects were associated with the administration of 4AR subcutaneously (via injections) in high doses to intact adult female Long-Evans rats over a variety of parameters. Twenty-one female Long-Evans rats were divided into three groups (age/body weight matched) at 50 days of age, n = 7 per treatment group: 1) dimethyl sufoxide (DMSO-vehicle) control, 2) 20 mg/Kg 4AR and 3) 90 mg/Kg 4AR. Then, the 4AR treatments were injected once a day for 21 consecutive days starting at 70 days of age (total volume per injection 0.1 cc). All animals were placed on a phytoestrogen-low diet (10 ppm) to eliminate dietary sources of phytoestrogens 2 weeks prior and throughout the experiment. Each rat was weighed daily, and food and water intake was monitored twice during the injection period (i.e. days 8-9 [amp] days 20-21). Before (60-68 days of age) and during the injection schedule the estrous cycle was monitored for eight days (days 10-18 during 4AR injections) in each animal. At 90 days of age, all rats were tested in the Porsolt forced-swim test (PFST) to index depressive-like behaviors. At 91 days of age all animals were sacrificed. Body weight was recorded, trunk blood, white adipose (WAT; measured to 0.1 grams) and hypothalamic [amp] pre-optic area tissues (for aromatase activity, fmol/hr/mg protein, via the tritiated water assay) were collected. The data was analyzed by ANOVA followed by pairwise comparisons (Tukey[apos]s), alpha = p [lt] 0.05. 4AR treatmens at 20mg/Kg or 90 mg/Kg had not effect on weight gain patterns, WAT deposition, hypothalamic aromatase activity, estrous cycle patterns, or depressive-like behaviors via the PFST. The obtained results suggest that 4AR is safe at high doses in intact female Long-Evans rats that have wide health applications, such as, anti-cancer, cardiovascular protection, anti-aging, anti-inflammatory, anti-oxidant and dermatological products.[br][br]Sources of Research Support: Funding providing, in part, by BYU TTO 19-221560 grant.[br][br]Nothing to Disclose: KF, CB, MA, EL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 509 292 1976 MON-26 PO31-02 Monday 1644 2012


1641 ENDO12L_MON-27 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Mitochondrial Steroidogenic Activity Requires Lipid-Mediated Unfolding of 3[beta]HSD2 Maheshinie Rajapaksha, James L Thomas, Himangshu S Bose Mercer University School of Medicine, Savannah, GA; Mercer University School of Medicine, Macon, GA; Mercer University School of Medicine, Savannah, GA Proteins conformation is determined by the energetic information specified within the sequence and involves a variety of intermediate states with decreasing free energies. Inner mitochondrial membrane (IMM) proteins that do not undergo cleavage of the N-terminal sequence, activity occurs in the absence of a mitochondrial receptor. For example human 3-beta hydroxysteroid dehydrogenase type2 (3[beta]HSD2), the IMM resident responsible for catalyzing two extremely important steps in steroid metabolism: the conversion of pregnenolone to progesterone and dehydroepiandrostene (DHEA) to Androstenedione. Conversion requires that 3[beta]HSD2 serves as a dehydrogenase and isomerase. The bifunctionality results from a conformational change, but the trigger of this change remains unknown. Using Fluorescence Resonance Energy Transfer (FRET), we found that 3[beta]HSD2 interacted with the charged phospholipid, Dipalmitoylphosphatidylglycerol (DPPG), and by doing so became less stable as indicated by the decrease in thermal denaturation (Tm), from 51.8[deg] C to 48[deg] C. The [alpha]-helical content decreased with the charged vesicles or a mixture of charged and uncharged (Dipalmitoylphosphatidylcholine, DPPC) vesicles without affecting the [beta]-sheet conformation. With the exception of the N-terminal 20 amino acids, mixed vesicles protected 3[beta]HSD2 from trypsin digestion. However, protein incubated with DPPC was only partially protected. The lipid-mediated unfolding completely supports the model in which a cavity forms between the [alpha]-helix and [beta]-sheet. Finally, the presence of charged vesicles enhanced 3[beta]HSD2 activity, suggesting that interaction with the lipid promotes the conformational changed needed for full enzyme activity. As 3[beta]HSD2 lacks a receptor, opening the conformation may stabilize the protein.[br][br]Sources of Research Support: National Institutes of Health HD057876 awarded to HSB.[br][br]Nothing to Disclose: MR, JLT, HSB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 258 292 1977 MON-27 PO31-02 Monday 1645 2012


1642 ENDO12L_MON-28 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Prepuberty: A More Vulnerable Target Period for Diethylhexyl Phthalate (DEHP) and Dibutyl Phthalate (DBP) To Affect Puberty Timing Compared to Neonatal Period Di Wu, Jialei Hu, Guizhen Du, Ling Song, Xinru Wang Institute of Toxicology, Nanjing Medical University, Nanjing, China; School of Public Health, Nanjing Medical University, Nanjing, China Epidemiological data showed that diethylhexyl phthalate (DEHP) and dibutyl phthalate (DBP) exposure may lead to precocious puberty. We hypothesized that besides neonatal period the time-window preceding puberty (prepuberty) may be a secondary target period for DEHP and DBP. To test our hypothesis newborn female Sprague-Dawley rats were cross-fostered and injected subcutaneously with three concentrations of DEHP or DBP (0.5mg/kg, 5mg/kg and 50 mg/kg) during neonatal period (postnatal day 1-5 P1-P5) and prepuberty (P25-P30) (10 female rats per group). We measured anogenital distance (AGD) and body weight at birth and on P35. AGD values were normalized to the cubed root of body weight for analysis. Ages of eye opening and vagina opening were recorded. Postpubertal estous cyclicity was examined based on vaginal cytology. Animals were perfused at proestrus following two complete cycles or on the 15th day after vagina opening. A blood sample was collected from the anesthetized rat just prior to the perfusion. Ovarian mass was measured and serum estrodial (E2), luteinizing hormone (LH) levels were detected. Effects of treatment and treating age on each endpoint were evaluated using two-way ANOVA followed by post hoc analysis when indicated by a significant interaction. The results showed that the interaction of treatment and treating age was significant on vagina opening age (p[lt]0.001), E2 level (p[lt]0.05), body weight on P35 (p[lt]0.001) and normalized AGD (p[lt]0.001). The main effects of treatment were significant on vagina opening age (p[lt]0.001), body weight on P35 (p[lt]0.001), ovarian weight/body weight index (p[lt]0.0001) and normalized AGD (p[lt]0.01). In detail, the DBP treatment at three concentrations all advanced vagina opening age. 50 mg/kg DBP group was heavier than the control. 5 mg/kg, 50 mg/kg DBP and 50 mg/kg DEHP groups had bigger ovarian weight/body weight index than the 0.5 mg/kg DEHP group. 5 mg/kg DEHP and control groups had longer normalized AGD than 0.5 mg/kg DEHP group. The main effects of treating age were significant on E2 level (p[lt]0.001), body weight on P35 (p[lt]0.05) and nearly significant on LH levels (p=0.055) with P26-P30 treating age groups having significantly higher E2 and LH levels, lower body weight. These data indicated that prepuberty is a target period during which reproduction maturation may be more vulnerable to DBP compared to neonatal period. It may be due to the less time to compensate the effects caused by the endocrine disruptors.[br][br]Sources of Research Support: National Natural Science Foundation of China 81102147.[br][br]Nothing to Disclose: DW, JH, GD, LS, XW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1202 292 1978 MON-28 PO31-02 Monday 1646 2012


1643 ENDO12L_MON-29 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Copy Number Variation (CNV) Polymorphisms in the [italic]CNTNAP4[/italic] and [italic]ESR1[/italic] Genes: Positive Association with Successful Aging Might Exclude the Carriers as Targets for Questionable Therapeutic Interventions Leonid Iakoubov, Malgorzata Mossakowska, Magdalena Owczarz, Monika Puzianowska-Kuznicka GeneCona, LLC, Vallejo, CA; International Institute of Molecular and Cell Biology, Warsaw, Poland; Medical Center of Postgraduate Education, Warsaw, Poland; Mossakowski Medical Research Centre, Warsaw, Poland [bold]BACKGROUND:[/bold] Last developments in HRT underscored the necessity to search for target sub-populations where therapy-related benefits will outweigh risks. Markers that are associated with successful aging might help to narrow down this search by the [italic]a priori[/italic] exclusion of their carriers from the pool of candidates for therapy as already having a rather beneficial prognosis. Here we investigated if genetic variants in [italic]CNTNAP4[/italic] and [italic]ESR1[/italic] genes could be such a markers.[br][bold]METHODS:[/bold] A report on female-specific association with aging for a CNV variant in the [italic]CNTNAP4[/italic](GC6del/del) in discovery and in replication studies is provided in a companion abstract (Iakoubov et al.) for general population and in the sub-population of carriers with a CNV variant in [italic]ESR1[/italic](GC2ins/ins). Here, the frequency of alternative variants of the two examined genes, GC6ins/ins and GC2del, were studied using TaqMan qPCR in female samples originated from Eastern European Caucasians. The frequency was compared between female age groups 65-75 (n=325) and 80-90 (n=311). Statistical significance was evaluated using Fisher[apos]s 2x2 exact test.[br][bold]RESULTS:[/bold] The GC2del variant of [italic]ESR1[/italic] was present in 30% of females age 65-75 and in 38% of age 80-90. The difference was statistically significant: OR=1.46, CI(1.04-2.02), Fisher exact P=0.02901.[br]The GC6ins/ins variant of [italic]CNTNAP4[/italic] was present in 13% of females age 65-75 and in 29% of age 80-90. The difference was highly statistically significant: OR=2.41, CI(1.62-3.58), Fisher exact P=0.00001.[br][bold]CONCLUSION:[/bold] Carriers of [italic]ESR1[/italic](GC2del) and [italic]CNTNAP4[/italic](GC6ins/ins) CNV variants have higher than average chances of successful aging till the age 80+. With such a beneficial prediction, these sub-populations should be excluded from being a potential target for therapeutic interventions like HRT.[br]This is specifically related to carriers of [italic]CNTNAP4[/italic](GC6ins/ins) variant who comprise about 13% among females aged [sim]60 years old. About two times elevated frequency of this variant among females aged 80+ justifies that all 65-75 years old female carriers of this genotype will successfully make it to 80+ with no need of innovative therapeutic interventions.[br][br]Nothing to Disclose: LI, MM, MO, MP-K 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 549 292 1979 MON-29 PO31-02 Monday 1647 2012


1644 ENDO12L_MON-30 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Regulation of Ghrelin Receptor mRNA Expression by Estradiol in the Hypothalamus of the Female Mouse Heather M Dungan Lemko, Charlotte E Lee, Jeffrey M Zigman, Joel K Elmquist, Carol F Elias University of Texas Southwestern Medical Center, Dallas, TX In the hypothalamus, the circuits controlling metabolism and fertility lie side by side and often overlap. Sex steroids derived from the gonads can act on hypothalamic neurons to alter energy storage and metabolism. Conversely, metabolic hormones produced by gut, pancreas, and adipose tissues act in the hypothalamus to regulate the function of the gonads. One such hormone is the peptide ghrelin. When reserves are low and a meal is expected, the stomach produces ghrelin which interacts with its receptor in the hypothalamus to stimulate feeding and mobilization of energy. Because previous studies have indicated that an estrogen-responsive element is present in the promoter region of the ghrelin receptor gene ([italic]GHSR[/italic]), we reasoned that the sex steroid estradiol could alter the effects of ghrelin signaling in the hypothalamus by regulating the expression of [italic]GHSR[/italic]. To test this theory, we first used [italic]in situ[/italic] hybridization to visualize the expression patterns of [italic]GHSR [/italic]mRNA in the brains of mice that were either ovariectomized to remove all sex steroids or treated with a chronic infusion of estradiol. Our initial observations were in accord with previously published expression patterns and we determined that, in most nuclei, [italic]GHSR[/italic] expression was not affected by chronic estradiol exposure. However, we did observe that [italic]GHSR[/italic] expression in the caudal arcuate nucleus of the hypothalamus (Arc) was robustly stimulated in the presence of estradiol. We then collected tissue punches from the caudal Arc and the anteroventral periventricular nucleus (AVPV) of ovariectomized or estradiol-treated mice. The AVPV was targeted along with the Arc because both nuclei are sites of [italic]GHSR[/italic] expression where estrogen receptors are also present. We used quantitative real-time PCR (qPCR) to determine the relative mRNA expression levels of [italic]GHSR[/italic] in these nuclei. As suggested by our [italic]in situ[/italic] data, qPCR analysis revealed a three-fold increase in [italic]GHSR[/italic] expression in the caudal Arc of estradiol-treated mice compared to mice not exposed to sex steroids. In contrast, [italic]GHSR[/italic] expression in the AVPV was not altered by the presence or absence of estradiol. This study suggests that sex steroids can regulate ghrelin signaling in specific regions of the hypothalamus, but further work is required to understand how estradiol and ghrelin signaling interact in the hypothalamus.[br][br]Sources of Research Support: NIDDK fellowship F32-DK085834; NICHD grant R01-HD61539; Foundation for Prader-Willi Research grant.[br][br]Nothing to Disclose: HMDL, CEL, JMZ, JKE, CFE 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 547 292 1980 MON-30 PO31-02 Monday 1648 2012


1645 ENDO12L_MON-31 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) Novel Maternal Neuropeptides: Amylin and Tuberoinfundibular Peptide 39 (Parathyroid Hormone 2) Arpad Dobolyi Semmelweis University and the Hungarian Academy of Sciences, Budapest, Hungary We recently detected amylin, a previously known pancreatic peptide in the maternal preoptic area (1). Amylin-containing neurons were activated by pup exposure and preoptic amylin levels correlated with maternal behavior in rats (2). In the present study, we investigated the interaction of amylin and tuberoinfundibular peptide 39 (TIP39). The expression of TIP39 has been described in 2 different sites in the brain (3). Recently, we identified an additional site of TIP39 expression in the posterior intralaminar complex of the thalamus (PIL) where TIP39 expression is significant only in postpartum mothers. A function of TIP39 in the regulation of prolactin release has been suggested. An antagonist of the parathyroid hormone 2 receptor, the receptor of TIP39, effectively eliminated suckling-induced prolactin release (4). In the present study, we report that in addition to the arcuate, peri- and paraventricular nuclei involved in the regulation of prolactin release, TIP39 neurons in the PIL also project to the medial preoptic area, an area whose lesion leads to the absence of maternal behaviors. A retrograde tracer (cholera toxin beta subunit) injected into the medial preoptic area labeled TIP39 neurons in the PIL. In turn, an anterograde tracer (biotinyliated dextran amine) injected into the PIL labeled fiber terminals in the preoptic area. The characteristic distribution pattern of anterogradely labeled fibers in the medial preoptic nucleus, parts of the medial preoptic area and the ventral subdivision of the bed nucleus of the stria terminalis correlated with that of amylin neurons. This distribution pattern was also similar to that of TIP39 and the parathyroid hormone 2 receptor in the preoptic area. Results of double labeling immunohistochemical and in situ hybridizational studies suggest that amylin neurons receive input from the PIL, which may be a thalamic relay center distributing suckling-related information towards different maternal centers of the hypothalamus.[br][br](1) Dobolyi A, Central amylin expression and its induction in rat dams. J Neurochem 2009; 111:1490-1500. (2) Szabo ER, Cservenak M, Dobolyi A, Amylin is a novel neuropeptide with potential maternal functions. FASEB J. 2011; Sep 29. Epub ahead of print. (3) Dobolyi A, Ueda H, Uchida H, Palkovits M, Usdin TB, Anatomical and physiological evidence for involvement of TIP39 in nociception. Proc Natl Acad Sci USA 2002; 99:1651-6. (4) Cservenak M, Bodnar I, Usdin TB, Palkovits M, Nagy GM, Dobolyi A, Tuberoinfundibular peptide of 39 residues is activated during lactation and participates in the suckling-induced prolactin release. Endocrinology 2010; 151:5830-40.[br][br]Sources of Research Support: Bolyai Grant of the Hungarian Academy of Sciences; OTKA NNF85612 and K100319 research grants awarded to AD.[br][br]Nothing to Disclose: AD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 19 292 1981 MON-31 PO31-02 Monday 1649 2012


1646 ENDO12L_MON-32 POSTER SESSION: Ovarian Regulation [amp] PCOS (1:30 PM-3:30 PM) A Novel 17-[beta]-Estradiol Prodrug Restores Diurnal Tail Skin Temperature Rhythm in Ovariectomized Rats: Potential Use To Treat Menopausal Hot Flushes Istvan Merchenthaler, Malcolm Lane, Katalin Prokai-Tatrai, Laszlo Prokai University of Maryland, Baltimore, MD; University of North Texas Health Science Center at Fort Worth, Fort Worth, TX; University of North Texas Health Science Center at Fort Worth, Forth Worth, TX [bold][italic]Background:[/italic][/bold] The major menopausal symptom women seek help for are hot flushes. Estrogen therapy has been the treatment of choice to alleviate these symptoms; however, many women cannot take estrogens. Therefore, there is a huge unmet need for novel, efficacious, and safe therapies. Para-quinol of 17[beta]-estradiol (DHED, based on its chemical structure) is selectively converted to 17[beta]-estradiol (E2) in the rodent brain, however, such conversion does not occur in the peripheral tissue including the uterus, breast, or the pituitary gland. Thus, DHED[apos]s action is restricted to the brain. Currently, only two animal models of hot flushes are available. One is the ovariectomized (OVX) morphine-addicted rat model in which estradiol and DHED reduce the morphine withdrawal-induced tail skin temperature (TST) rise. The other model is based on OVX-induced loss of TST decreases during the dark phase. Estrogens have been shown to maintain a rhythmic diurnal rhythm of TST in OVX rats similarly to sham-operated rats (Berendsen et al. 2001; Deecher et al. 2007). The current studies were designed to test whether DHED can also restore diurnal rhythm of TST, i.e., decrease TST in the dark phase in OVX rats.[br][bold][italic]Methods:[/italic][/bold] Rats were OVX-ed and implanted with a telemetric device sc with its thermistor just below the skin of the tail. Rats were allowed to recover for one week before TST was measured continuously. Baseline was measured for one week, then, the animals were treated orally either with DHED (50 ug/kg), ethynyl estradiol (EE, 200 ug/kg) or vehicle for another week. At the end of the experiments, rats were euthanized and their brains, uteri and pituitary glands removed and processed for [italic]in situ[/italic] hybridization and RT-PCR to evaluate the expression of estrogen-regulated genes in the pituitary and uterus.[br][bold][italic]Results:[/italic][/bold] Similarly to EE, DHED restored the OVX-induced loss of diurnal TST rhythm to that seen in intact, cycling rats. The effect of EE and DHED was rapid, i.e., following the first dosing in the morning, TST in the dark phase was already reduced. Importantly, DHED, contrary to EE, did not stimulate the uterus (wet weight) or the expression of galanin in the anterior pituitary while stimulating the expression of progesterone receptor in the preoptic area of the hypothalamus.[br][bold][italic]Conclusion: [/italic][/bold]The use of a CNS-specific DHED may provide a novel and promising treatment for women who suffer from menopausal hot flushes without causing side effects in the periphery including the uterus.[br][br]1. Merchenthaler I, Funkhouser JM, Carver JM, Lundeen SG, Ghosh K, Winneker RC. Maturitas 30:307-316, 1998. 2. Berendsen HHG, Weekers AHJ, Kloosterboer HJ. Eu J Pharm 419:47054, 2001. 3. Deecher DC, Alfinito PD, Leventahl L, Cosmi S, Johnston GH, Merchenthaler I, Winneker R. Endocrinology 148:1376-1383, 2007.[br][br]Sources of Research Support: NIH grants 1RO1AG031535-01A2, 3R01AG031535-01A2S1, and AG031535.[br][br]Nothing to Disclose: IM, ML, KP-T, LP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1860 292 1982 MON-32 PO31-02 Monday 1650 2012


1647 ENDO12L_MON-33 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Comparison of Mass Spectrometry vs Immunoassay for Testosterone, and Correlation with Dihydrotestosterone, Estradiol and Other Characteristics in a Population-Based Cohort of Older Men Bu Beng Yeap, Helman Alfonso, Stephen A Paul Chubb, David J Handelsman, Leon Flicker University of Western Australia, Perth, Australia; Fremantle Hospital, Fremantle, Australia; University of Western Australia, Perth, Australia; University of Western Australia, Perth, Australia; Royal Perth and Fremantle Hospitals, Perth, Australia; University of Sydney, Sydney, Australia Introduction[br]Reduced circulating testosterone (T) may be a causative factor for health problems in older men, or an epiphenomenon. Immunoassays for T are imprecise, and the roles of dihydrotestosterone (DHT) and estradiol (E2) are even less clear. We compared T results from two assays and the relationship of T with DHT and E2 in older men.[br]Methods[br]We studied community-dwelling men aged 70-89 years, who had early morning blood sampling. T was measured by immunoassay (IA) (Immulite 2000, Diagnostic Products Corp) and by liquid chromatography-tandem mass spectrometry (LC-MS) using a previously validated method [1]. DHT and E2 were measured by LC-MS [1].[br]Results[br]In 4,037 men aged (mean[plusmn]SD) 75.3[plusmn]4.1 years, mean T measured by IA was 15.2[plusmn]5.9 nmol/L compared with LC-MS 12.9[plusmn]5.1 nmol/L (mean difference +21%). There was greater bias towards the lower end of the range of T values. Exclusion of men receiving androgens or anti-androgen therapy, and those reporting orchidectomy or prostate cancer, left 3,692 men for further analysis. Using IA, 5.2% of men had T[lt]8 nmol/L, compared to 11.2% with LC-MS. Using LC-MS, men with T[lt]8 nmol/L had lower levels of DHT (0.83 vs 1.58 nmol/L, difference=-0.75, 95%CI=-0.81,-0.70) and E2 (56.6 vs 77.8, difference=-21.2, 95%CI=-23.5,-19.0) vs men with T[ge]8 nmol/L. LC-MS T was moderately correlated with E2 (r=0.48) and DHT (r=0.69). In multivariate regression models, age (odds ratios [OR]=1.22, 1.60 and 1.88 for 75-79, 80-84 and 85+ yrs vs [le]74, all p[lt]0.05), BMI (OR=2.07 and 4.29 for 25-30 and [gt]30 vs [le]25 kg/m[sup]2[/sup], all p[lt]0.001), never-smoking status (OR=1.40, p[lt]0.001) and diabetes (OR=1.91, p[lt]0.001) were associated with T in the lowest quintile of values. Age (OR=1.44, 1.92 for 75-79, 80-84 vs [le]74 yrs, all p[lt]0.001), BMI (OR=1.84, 2.93 for 25-30, [gt]30 vs [le]25 kg/m[sup]2[/sup], all p[lt]0.001) and diabetes (OR=1.96, p[lt]0.001) were associated with lower DHT. Only age (OR=1.30, 1.55 for 75-79, 80-84 vs [le]74 yrs, all p[lt]0.001) and diabetes (OR=1.34, p[lt]0.01) were associated with lower E2.[br]Conclusions[br]Compared with LC-MS, this method for IA overestimated T in older men. LC-MS should be used to estimate accurately the prevalence of low T in ageing men. While low levels of T are associated with lower DHT and E2, there is limited correlation of E2 with T. Diabetes is associated with lower T, DHT and E2, while BMI is associated with lower T and DHT but not E2. Additional studies are needed to clarify whether reduced DHT or E2 levels predict health outcomes independently of T.[br][br][1] Harwood DT, Handelsman DJ. Development and validation of a sensitive liquid chromatography[ndash]tandem mass spectrometry assay to simultaneously measure androgens and estrogens in serum without derivatization. Clinica Chimica Acta 2009; 409: 78-84.[br][br]Sources of Research Support: This work was supported by funding from the National. Health and Medical Research Council (NHMRC) of Australia Project Grants 279408, 379600, 403963, 513823 and 634492), and by a Clinical Investigator Award to B.B.Y. from the Sylvia and Charles Viertel Charitable Foundation, New South Wales, Australia.[br][br]Nothing to Disclose: BBY, HA, SAPC, DJH, LF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 886 293 1983 MON-33 PO36-01 Monday 1651 2012


1648 ENDO12L_MON-34 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Longitudinal Changes in Testosterone over 5 Years in Community-Dwelling Men Andre B Araujo, Zumin Shi, Sean A Martin, Gary A Wittert New England Research Institutes, Inc, Watertown, MA; University of Adelaide, Adelaide, Australia There are few population-based studies reporting longitudinal changes in testosterone (T) levels, and limited information on risk factors for longitudinal change in T. We examined changes in serum T levels assessed by liquid chromatography mass spectrometry (LC-MS/MS) among a cohort of 1,588 men aged 35-80y at baseline with available data at 2 clinic visits separated by 5 years. Excluded from analyses were men on medications (N=102) or with medical conditions (N=45) known to affect hormones, and men with pathological laboratory values (N=59), leaving 1,382 for analysis. Multivariate linear regression was used to assess the association between baseline predictors as well as change in predictors and annualized change in T. All models were minimally adjusted for baseline age. At baseline, mean age was 54[plusmn]11y, 21% were unmarried, 19% were current smokers, 30% had BMI[ge]30kg/m[sup]2[/sup], 48% had waist[ge]100cm, and 8% were depressed. Mean baseline T was 16.2[plusmn]1.4nmol/L, with 3%[lt]8nmol/L, 36% 8-15nmol/L, and 61%[gt]15nmol/L. Mean T at follow-up was 15.6[plusmn]1.4nmol/L, representing a non-significant T change of -0.13nmol/L/y (95% CI: -0.62, 0.41) or an approximate decline of -0.80%/y. Baseline variables significantly associated with annualized T changes were: being unmarried (-0.18nmol/L/y vs. married, p[lt].01), central obesity (+0.13nmol/L/y vs. non-central obesity, p[lt].01), BMI[ge]30kg/m[sup]2[/sup] (+0.22nmol/L vs. [lt]25kg/m[sup]2[/sup], p[lt].01), and current smoking (-0.17nmol/L/y vs. non-smoker, p[lt].01). The effects of smoking status was driven by decreasing T in men who quit smoking (-0.17nmol/L/y vs. non-smoker at both time points, p[lt].01). The effects of central obesity and high BMI were driven by changes in status between baseline and follow-up, with for instance, a T change of -0.25nmol/L/y in men who became centrally obese (waist[ge]100cm) and a T change of +0.20nmol/L/y in men who became non-centrally obese, both compared to men who were not centrally obese at both time points (both p[lt].01). In addition, men who were depressed at both time points had a significantly greater T decline (-0.28nmol/L/y vs. men without depression at both time points, p[lt].01). In conclusion, this study strongly suggests that (i) T decline is not an inevitable part of aging and (ii) variability in T changes are largely explained by smoking behavior and intercurrent changes in health status, particularly obesity and depression.[br][br]Sources of Research Support: This study was supported by the National Health and Medical Research Council of Australia (NHMRC grant 627227).[br][br]Nothing to Disclose: ABA, ZS, SAM, GAW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1960 293 1984 MON-34 PO36-01 Monday 1652 2012


1649 ENDO12L_MON-35 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Low Testosterone as an Independent Predictor of Mortality in Men with Chronic Liver Disease Mathis Grossmann, Rudolf Hoermann, Linsey Gani, Irene Chan, Ada Cheung, Jeffrey Zajac, Paul Gow, Peter Angus University of Melbourne, Heidelberg, Australia; Austin Health, Heidelberg, Australia; Austin Health, Heidelberg, Australia Objective: To examine the prevalence and prognostic implications of low serum testosterone in men with chronic liver disease.[br]Design: We conducted an observational study at a tertiary referral centre.[br]Patients and measurements: Baseline serum testosterone was measured in 171 men presenting to the Victorian Liver Transplant Unit for liver transplant evaluation. Patients were followed to liver transplant, or death.[br]Results: Sixty-one percent of men had a low total testosterone (TT, [lt]10 nmol/L), and 90% of men had a low calculated free testosterone (cFT, [lt]230 pmol/L). During the available observation time (median 8 months, interquartile range 4-14 months), 56 men (33%) died, and 63 (37%) received a liver transplant. Fifty-two (30%) survived without a transplant. Median time to death was 8 months (range 2-13), and to liver transplant 8 months (4-14). Baseline low TT and cFT both (p[lt]0.0001) predicted mortality. Moreover, in a Cox proportional hazard model, both low total (p= 0.02) and free testosterone (p= 0.007) remained predictive of death independently of established prognostic factors, such as the Model for End Stage Liver Disease (MELD) score, and serum sodium levels. A decrease in TT by 1 nmol/L and in cFT by 10 pmol/L was associated with an 8% increase in mortality.[br]Conclusions: Low testosterone levels are common in men with severe liver disease and predict mortality independent of MELD, the standard score used to prioritize the allocation of liver transplants.[br][br]Sources of Research Support: This work was supported by grants from the National Health and Medical Research Council of Australia ((#400417, #1006407).[br][br]Nothing to Disclose: MG, RH, LG, IC, AC, JZ, PG, PA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 140 293 1985 MON-35 PO36-01 Monday 1653 2012


1650 ENDO12L_MON-36 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Frequent Occurrence of Hypogonadism in Male Veterans with Type 2 Diabetes Mellitus Sheila Pinkson, Zandra Perez-Cadena, Maureen Koops, Devjjit Tripathy University of Texas Health Science Center, San Antonio, TX; ALM VA Hospital, San Antonio, TX [bold]Background and aims[/bold]: Although testosterone levels decline by 0.5-2% per year after 3[sup]rd[/sup] decade, the effect of T2DM on testosterone levels is not clear. Given the recent general awareness of low testosterone levels in the United States, the aim of this study was to characterize the prevalence of low testosterone levels in a relatively homogeneous population of middle aged male US veterans with T2DM. [bold]Methods[/bold]: We examined the prevalence of low testosterone levels in 113 male veterans, 95 participating in the VA Diabetes trial and 18 T2DM subjects attending the VA diabetes clinic who had total and free testosterone, LH, FSH, and prolactin levels analyzed. Based on VA laboratory standards hypogonadism was defined as total testosterone [lt]300ng/dl.[br][bold]Results:[/bold] The mean age was 54 [plusmn] 0.6yrs (range 34 to 65yrs), BMI was 31.8 [plusmn] 0.5 kg/m[sup]2[/sup] and mean HBA[sub]1c[/sub] was 8.2 [plusmn] 0.1%. The mean total testosterone was 370 [plusmn]17 ng/dl and mean free testosterone was 10.7 [plusmn]0.4 ng/dl. Forty-one percent (45/113) of patients hypogonadal with total testosterone levels [lt]300ng/dl and 21 (19%) had testosterone levels [lt]200ng/dl. Of those patients with low testosterone, 74% had low/normal FSH (5.6[plusmn]0.5) and 26% had elevated FSH (17.1[plusmn]0.4). Amongst subjects with normal total testosterone levels, 24% had elevated FSH (13.3[plusmn]0.8) or subclinical hypogonadism. There was an inverse correlation between BMI and total testosterone (r = -0.443; p[lt]0.005) as well as with free testosterone (r=-0.383; p[lt]0.005). When subjects were stratified into quartiles of BMI, plasma testosterone levels progressively declined with increasing BMI (p[lt]0.05 for trend). On the other hand, when subjects were classified into 4 age groups- [lt]50 yr, 50-54yrs, 55-59 and [gt]60 yrs, no such trend in testosterone levels were observed. The total testosterone correlated with hematocrit levels (r = 0.197; p = 0.04). There was no relationship between age (r=0.138, p=ns), duration of diabetes (r=0.25,p=ns) or HbA[sub]1c[/sub] levels (r=0.053,p=ns) and total/free testosterone.[br][bold]Conclusion:[/bold] The prevalence (41%) of hypogonadism in veterans with T2DM much greater than the age related decline in testosterone levels in non-diabetic individuals. Hypergonadotropic hypogonadism (26%) and subclinical hypogonadism (24%) were also frequently seen. Increasing BMI was the most important predictor of low testosterone levels in veterans. Age, duration of diabetes and glycemic control do not impact the total testosterone levels in T2DM.[br][br]Sources of Research Support: ALM VA Hopspital , Bartter Research Unit. NIH.[br][br]Nothing to Disclose: SP, ZP-C, MK, DT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1369 293 1986 MON-36 PO36-01 Monday 1654 2012


1651 ENDO12L_MON-37 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Low Sex Hormone Binding Globulin and Testosterone Associated with Increased Risk of Metabolic Syndrome in Non-Obese Asian Men, but Not Obese Asian Men Troy Hai Puar, Joan Joo Khoo, Yuan Tud Chen Changi General Hospital, Singapore, Singapore Introduction:[br]Lower concentrations of sex hormone binding globulin (SHBG) and serum total testosterone (TT) are associated with increased risk of metabolic syndrome in Caucasian men. We studied if this association similarly affects both obese and non-obese Asian men.[br]Methods:[br]We conducted a cross-sectional study of 353 Asian men, 90.9% Chinese, mean age 51.6[plusmn]9.3 years. Serum levels of TT, SHBG, triglycerides (TG), high-density lipoprotein (HDL), fasting plasma glucose (FPG) were measured, along with waist circumference (WC) and body mass index (BMI). Metabolic syndrome (MetS) was defined using the National Cholesterol Education Program Adult Treatment Panel III Criteria for Asian population with WC [ge] 90cm. Obesity was defined for the Asian population as BMI [gt]28kg/m2.[br]Results:[br]In the entire study population, univariate analysis (after adjusting for age) showed that both SHBG and TT were inversely associated with increased risk of MetS, odds ratio (OR) 0.96 (95% CI: 0.94-0.98, p[lt]0.01) and 0.90 (0.86-0.94, p[lt]0.01) respectively. A similar association was found in non-obese Asian men in whom the OR of SHBG and TT in relation to MetS was 0.97 (95% CI: 0.95 [ndash] 0.99, p=0.03) and 0.93 (95% CI: 0.87 [ndash] 0.99, p=0.02) respectively. However, in obese men, no significant association was found between either SHBG or TT and MetS, OR 0.99 (95% CI: 0.96 [ndash] 1.02, p=0.47) and 0.99 (95% CI: 0.91 [ndash] 1.08, p=0.84).[br]Conclusion:[br]While low SHBG and TT have been associated with increased risk of MetS in Caucasian men, this has not been studied in an Asian population. Furthermore, we found an association of low SHBG and TT with increased risk of MetS in non-obese Asian men, but not obese Asian men, emphasizing the importance of assessing metabolic risk even in non-obese men with low SHBG and TT. Future prospective studies need to be done to demonstrate the effect of low SHBG and TT in increasing the risk of developing MetS in Asian men with normal weight.[br][br]Nothing to Disclose: THP, JJK, YTC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1429 293 1987 MON-37 PO36-01 Monday 1655 2012


1652 ENDO12L_MON-38 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Epidemiology of Diagnosed Hypogonadism and Testosterone Use in a Nationally Representative Study of Ambulatory Care Andre B Araujo, May H Yang, Varant Kupelian, Susan A Hall, Deborah C Brander, Kishore L Lakshman New England Research Institutes, Inc, Watertown, MA; Prima-CARE Medical Center, Fall River, MA; Partner[apos]s Health Care Inc, Boston, MA Little is known about the epidemiology of hypogonadism (HG) and testosterone (T) use in broad, unselected populations. We examined the epidemiology of HG and T use among men [ge]18y using weighted patient clinic visit data collected in the National Ambulatory Medical Care Survey, a national probability survey of US office-based physicians. Binary outcomes were HG (ICD-9: 257.1-257.9) and prescription T. The relationships of outcomes to demographics and comorbidity were quantified with odds ratios (OR) and 95% confidence intervals (CI) obtained via logistic regression models (minimally adjusted for age). The complex sampling design was accounted for by using sampling weights in the analysis. The analysis sample included 49,469 patient visits (mean age: 56y), which represented over 1.5 million visits to ambulatory care by adult men in the US from 2005-09. There were 147 visits (0.3%) with a code of HG, with most (144 or 96%) coded as ICD-9 257.2. The 147 visits represent a weighted count of 4,900 visits coded with HG. T was prescribed at 260 (0.5%) visits (weighted N=8,196). Gel formulations increased significantly (P=.005) over time, displacing injectable formulations with no net change (p=.06) in T use overall. Of the 147 visits with HG, 90 (55%) received a T prescription. T was prescribed in 170 (0.4%) visits without a code for HG. A code for HG was more likely in obese patient visits (adjusted OR=2.20, 95% CI: 1.07-4.51) or diabetes patient visits (adjusted OR=1.89, 95% CI: 1.09-3.26) but less likely in cancer patient visits (adjusted OR=0.30, 95% CI: 0.11-0.82). T prescription was more likely in patient visits with obesity (adjusted OR=1.97, 95% CI: 1.08-3.59), depression (adjusted OR=2.00, 95% CI: 1.19-3.37), and hyperlipidemia (adjusted OR=1.72, 95% CI: 1.17-2.53). The frequency of HG and T use in US ambulatory care is relatively low and appears to vary among patients with and without medical comorbidity. These estimates of [ldquo]visit prevalence[rdquo] are limited by the fact that they are based on visits (not patients), reliance on ICD-9 codes (vs. blood sampling/symptom assessment), and the fact that only up to 3 diagnoses were codable. T was prescribed at many visits without documentation of HG, which may represent undercoding of HG in clinical practice. T was prescribed in half of the visits where HG was coded, which when compared to the limited number of prior studies, is a relatively sizeable proportion.[br][br]Sources of Research Support: Abbott Laboratories.[br][br]Nothing to Disclose: ABA, MHY, VK, SAH, DCB, KLL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2020 293 1988 MON-38 PO36-01 Monday 1656 2012


1653 ENDO12L_MON-39 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Physician-Related Characteristics and Health Care Utilization in Association with Diagnosed Hypogonadism and Testosterone Use in a Nationally Representative Study of Ambulatory Care Andre B Araujo, May H Yang, Varant Kupelian, Susan A Hall, Deborah C Brander, Kishore M Lakshman New England Research Insistitutes, Inc, Watertown, MA; Prima-CARE Medical Center, Fall River, MA; Partner[apos]s Health Care Inc, Boston, MA Our understanding of physician characteristics and health care utilization (HCU) patterns associated with the diagnosis of hypogonadism (HG) or testosterone (T) prescription is limited. We described a large set (N=18) of physician-related characteristics and HCU variables in patient visits where a diagnosis of HG was made or where T was prescribed among men [ge]18y in the US using weighted patient clinic visit data collected from 2005-09 in the National Ambulatory Medical Care Survey, a national probability survey of US office-based physicians. Binary outcomes were HG (ICD-9: 257.1-257.9) and prescription T. The relationships of exposures to outcomes were quantified with odds ratios (OR) and 95% confidence intervals (CI) obtained via logistic regression models (minimally adjusted for age and number of comorbidities). The complex sampling design was accounted for by using sampling weights in the analysis. The analysis sample included 49,469 patient visits (mean age: 56 y) during the study period. There were 147 visits (0.3%) with a code of HG and 260 (0.5%) with T prescriptions. T was prescribed most often in visits to general and family practice (33%) and [ldquo]other[rdquo] specialties (29%), followed by internal medicine (24%) and urology (16%). Of the factors examined, only PSA testing (adjusted OR=3.15, 95% CI: 1.70-5.84) and weight reduction counseling (adjusted OR=2.23, 95% CI: 1.02-4.86) were more likely to occur during visits where HG was diagnosed. Glucose (adjusted OR=2.59, 95% CI: 1.06-6.35) and urinalysis (adjusted OR=1.92, 95% CI: 1.15-3.23) testing were more likely and tobacco education (adjusted OR=0.12, 95% CI: 0.03-0.57) was less likely to occur during visits where T was prescribed. Although PSA testing was more common in visits with T prescriptions, it was not significant (adjusted OR=1.61, 95% CI: 0.83-3.10). Physician specialty, time spent with the physician, and other screening tests (e.g., rectal exam, cholesterol, imaging) or health education were not related to HG diagnosis or T prescription. In the ambulatory care setting, diagnosis of HG or T prescribing was associated with various HCU factors, although it is unclear whether diagnosis/prescribing in the context of HG is triggered by symptoms or coincident with laboratory testing for other reasons.[br][br]Sources of Research Support: Abbott Laboratories.[br][br]Nothing to Disclose: ABA, MHY, VK, SAH, DCB, KML 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1862 293 1989 MON-39 PO36-01 Monday 1657 2012


1654 ENDO12L_MON-40 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) The Association of Sleep Restriction and Sleep Quality with Sex Hormone Levels in Community-Dwelling Aging Men Elizabeth A Suarez, Rebecca P Piccolo, Gary A Wittert, Peter Y Liu, Donald L Bliwise, H Klar Yaggi, Andre B Araujo New England Research Institutes, Inc, Watertown, MA; University of Adelaide, Adelaide, Australia; Woolcock Institute of Medical Research, University of Sydney, Sydney, Australia; Emory University School of Medicine, Wesley Woods Health Center, Atlanta, GA; Yale University School of Medicine, New Haven, CT Experimental laboratory studies of young, healthy men have demonstrated a detrimental effect of sleep restriction on circulating waking testosterone (T) levels. Similar effects of reduced sleep quality and efficiency on T have also been suggested. The relationship between sleep and sex hormones has been given limited consideration in older community-dwelling men. We examined waking sex hormone levels in a population-based sample of men in relation to self-reported sleep habits. Data were obtained from the Boston Area Community Health (BACH) Survey, a population-based survey of 5,503 racially/ethnically diverse men and women. Men were aged 27-79 years at time of data collection (2002-2005). Those on hormone therapy or taking other medications that affect hormone levels were excluded from this analysis. Sex hormone levels were measured from non-fasting serum samples collected in the early morning. Sleep habits were obtained via self-report for two measures, restricted sleep and low sleep quality. Restricted sleep was defined as less than 5 hours per night (collected as a binary variable), and poor sleep quality was defined as restless sleep over the past week in the presence of prescription or over-the-counter sleep aids (collected by medication inventory). Generalized linear models were used to calculate adjusted geometric mean hormone levels, modeling continuous log-transformed sex hormone variables as the outcome with each dichotomous sleep predictor independently and controlling for potential confounders chosen by backward selection. All models were minimally adjusted for age and self-rated health, in addition to other demographic and clinical factors as necessary. The mean age of the 1,801 men ([plusmn]SEM) in the analysis was 46.5[plusmn]0.5 years. Mean hormone levels were 441.0[plusmn]8.2 ng/dL (total T), 9.3[plusmn]0.2 ng/dL (free T), 33.4[plusmn]0.6 nmol/L (SHBG), and 5.4[plusmn]0.1 IU/L (LH). Free T was 12% lower among those with restricted sleep (p=0.03) and 24% lower among those with poor sleep quality (p=0.04). No significant relationships were found between restricted sleep or low sleep quality and total T, SHBG, or LH after controlling for potential confounders. Poor sleep quality is an independent predictor of free T levels in community-dwelling men. Whether improvements in sleep duration and quality would beneficially affect hormone levels is unknown.[br][br]Sources of Research Support: This study was supported by awards R21MD006769 from the National Institute on Minority Health and Health Disparities and U01DK056842 from the National Institute of Diabetes and Digestive and Kidney Disorders. The content is solely the responsibility of the authors and does not necessarily represent the official views of NIMHD, NIDDK, or NIH.[br][br]Disclosures: DLB: Consultant, Ferring Pharmaceuticals. Nothing to Disclose: EAS, RPP, GAW, PYL, HKY, ABA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1586 293 1990 MON-40 PO36-01 Monday 1658 2012


1655 ENDO12L_MON-41 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Effect of Testosterone Replacement on Response to Sildenafil Citrate in Men with Erectile Dysfunction: A Randomized, Placebo-Controlled Trial Matthew Jonathan Spitzer, Shehzad Basaria, Maithili N Davda, Thomas G Travison, Amanda Paley, Beth Kaplan, Norman A Mazer, Philip E Knapp, Samson Hanka, Jagadish Ulloor, Anqi Zhang, Katie Orwoll, Richard Eder, Lauren Collins, Nurahmed Mohammed, Shalender Bhasin Boston University School of Medicine and Boston Medical Center, Boston, MA; Boston University School of Public Health, Boston, MA; Hoffman LaRoche, Basel, Switzerland [bold]Background:[/bold] Erectile dysfunction (ED) and low testosterone levels frequently co-occur in men. It is unclear whether the addition of testosterone to a phosphodiesterase-5-inhibitor improves erectile response.[br][bold]Methods:[/bold] In this randomized, double-blind, placebo-controlled trial, men age 40 to 70 years old with Erectile Function Domain (EFD) scores of [le]25 on the International Index of Erectile Function (IIEF), total testosterone [lt]330ng/dL (measured by LC-MS/MS) and/or free testosterone [lt]50pg/mL, were randomized to either 10-g testosterone or placebo gel for 14-weeks after they had received sildenafil for a period of 3-7 weeks. The primary outcome was change in EFD score. Secondary outcomes included changes in other domains of the IIEF and Male Sexual Health Questionnaire (MSHQ), frequency of total and successful sexual encounters, ED-Quality-of-Life (ED-QOL), marital-intimacy, vitality, and affectivity-balance. Sample mean changes and 95% confidence intervals (CI) were computed, and a Student[apos]s T-test was used to assess for differences by treatment arm.[br][bold]Results:[/bold] Testosterone and placebo groups were similar at baseline. Mean serum total testosterone at screening was 248ng/dL and 254ng/dL, which increased to 649ng/dL and 338ng/dL on sildenafil plus testosterone and sildenafil plus placebo, respectively. Administration of sildenafil alone was associated with substantial increases in EFD score (mean change for both arms 7.7; 95%CI 6.5 to 8.8) as well as in other domains of IIEF, MSHQ, frequency of total and successful sexual encounters, and ED-QOL. However, after randomization, the change in EFD score did not differ significantly between testosterone plus sildenafil (mean change 1.7) and placebo plus sildenafil (mean change 0.4; difference 1.3; 95%CI -0.8 to 3.5; [italic]P[/italic]=0.23) arms. Similarly, changes with testosterone vs. placebo gel in other domains of the IIEF, frequency of total and successful sexual encounters, ED-QOL, marital-intimacy, vitality, and affectivity-balance did not differ between the two groups. Sensitivity analyses of men with baseline testosterone [lt]250ng/dL did not reveal significant differences in EFD scores between arms.[br][bold]Conclusions:[/bold] Combined administration of testosterone plus sildenafil was not superior to sildenafil plus placebo in improving erectile function in men with ED and low testosterone levels.[br][br]Nothing to Disclose: MJS, SB, MND, TGT, AP, BK, NAM, PEK, SH, JU, AZ, KO, RE, LC, NM , SB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1750 293 1991 MON-41 PO36-01 Monday 1659 2012


1656 ENDO12L_MON-42 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Cardiovascular Safety and Testosterone Replacement Therapy in Male Hypogonadism Including Men with Type 2 Diabetes and Cardiovascular Disease Jonathan Charles Brooke, David Stuart McLaren, Deborah J Walter, Vakkat Muraleedharan, Thomas Hugh Jones Barnsley Hopsital, NHSFT, Barnsley, UK; University of Sheffield, Sheffield, UK Testosterone replacement therapy (TRT) has recently been shown to have beneficial effects on CVD risk factors including insulin resistance, dyslipidaemia, obesity, hypertension and pro-atherogenic states. Nevertheless, the use of TRT remains controversial in men with cardiovascular disease.[br]A long-term retrospective audit of 401 (47.4% Type 2 diabetes (T2D), 34.0% CVD) hypogonadal men (age 58.7[plusmn]14.3years; mean baseline testosterone 7.16[plusmn]2.59nmol/l) receiving physiological TRT (84% testosterone gels) for up to 27 years (mean 5.41 years) in normal clinical practice was carried out to evaluate TRT[apos]s safety. BMI, waist circumference, blood pressure (BP), hemoglobin (Hb), hematocrit (HCT), lipid profile, liver function, testosterone, estradiol and PSA levels were monitored at 3, 6 and 12 months and yearly thereafter. Data from the most recent visits represent the primary endpoint. Hospital admissions, major adverse cardiovascular events (MACEs), mortalities and prostate-related outcomes were recorded.[br]TRT was associated with a reduction in total cholesterol (-0.28mmol/l, p=0.004), non- fasting triglycerides (-0.244mmol/l, p=0.04) and liver transaminases (ALT and AST) at the primary endpoint. HDL cholesterol fell by 0.053mmol/l (p=0.044). Hb increased by 0.72g/dl (p=0.002), HCT by 0.035 (p=0.011), PSA by 0.26g/l (p=0.008) and estradiol 128.45[plusmn]44.58 at baseline vs 157[plusmn]66.89pmol/l. At the end of the study PSA levels were comparable to those of eugonadal males and were not grossly elevated. No significant changes in BMI, waist circumference or BP were observed. Safety outcomes: Polycythaemia (HCT [gt]0.52) developed in 22 patients. 3 new cases of BPH and 2 prostate carcinomas were diagnosed, but this is no more than expected in this population which included aging males. 4 patients were diagnosed with T2D during the study, and 24 MACEs (4 MI, 8 angina, 5 TIAs, 2 CVAs, 4 CABG, 1 CCF) were identified. Over the course of the study there were 54 hospital admissions and 2 deaths which are no higher than expected in this morbid population.[br]This audit confirms the cardioprotective role of physiological doses of testosterone and demonstrates that in clinical practice TRT has beneficial effects on cardiovascular risk factors including circulating lipid and cholesterol levels. This is the largest and most comprehensive study of TRT safety to date, representing over 1642 patient years of TRT. TRT was not associated with an increase in prostate carcinoma, MACEs or mortality.[br][br]Nothing to Disclose: JCB, DSM, DJW, VM, THJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2226 293 1992 MON-42 PO36-01 Monday 1660 2012


1657 ENDO12L_MON-43 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Effect of Testosterone Administration on Hepatic Fat and Insulin Resistance in Older Men in a Placebo-Controlled Randomized Trial Grace Huang, Shalender Bhasin, Elizabeth Tang, Adam Aakil, Stephan Anderson, Hernan Jara, Maithili Davda, Thomas Travison, Shehzad Basaria Boston University School of Medicine, Boston, MA [bold]Background: [/bold]Non-alcoholic fatty liver disease is highly prevalent and is associated with insulin resistance and proatherogenic dyslipidemia. Androgen receptor knock-out male mice display hepatic steatosis and insulin resistance, suggesting that AR signaling may regulate hepatic fat. However, the effects of testosterone on hepatic fat in men are unknown.[br][bold]Objective: [/bold]To determine the effects of 6 months of testosterone administration on hepatic fat in older men with mobility limitation and low serum total and free testosterone levels who were participating in a placebo-controlled randomized trial (The TOM Trial).[br][bold]Methods: [/bold]209 men were randomized in the parent trial to either placebo or 10 g testosterone gel daily for 6 months. Magnetic resonance imaging (MRI) scans of the liver were obtained in 73 men (36 in the placebo and 37 in the testosterone group; mean age 74 years), and hepatic fat was determined using volumetric method. Fasting glucose and insulin levels were measured and HOMA[sub]IR[/sub] was calculated.[br][bold]Results: [/bold]Baseline liver volumes (1583 [plusmn] 363 ml vs 1522 [plusmn] 271 ml, p= 0.42), age, BMI, HOMA[sub]IR[/sub], fasting lipid profile and liver function tests were not significantly different in the testosterone and placebo arms. Serum testosterone concentrations increased from 250 [plusmn] 72 ng/dl and 229 [plusmn] 58 ng/dl to 632 [plusmn] 363 ng/dl and 272 [plusmn] 153 ng/dl in the testosterone and placebo groups, respectively. After 6 months of testosterone administration, the changes in liver volume were not different between the two groups (p-value = 0.5). Similarly, the change in HOMA[sub]IR[/sub] was not significantly different between the groups. Multiple regression analysis did not show any relationship between the change in testosterone concentrations and change in liver volume.[br][bold]Conclusion: [/bold]Testosterone administration in older men with mobility limitation and low testosterone was not associated with a reduction in hepatic fat. Larger clinical trials are needed to determine whether testosterone reduces liver fat and outcomes in men with nonalcoholic hepatic steatosis.[br][br]Nothing to Disclose: GH, SB, ET, AA, SA, HJ, MD, TT, SB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1704 293 1993 MON-43 PO36-01 Monday 1661 2012


1658 ENDO12L_MON-44 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Long-Term Effects of Testosterone Replacement Therapy on Cardiovascular Risk Factors in Hypogonadism, Including Men with Cardiovascular Disease and/or Type 2 Diabetes David Stuart McLaren, Jonathan Charles Brooke, Deborah J Walter, Vakkat Muraleedharan, Thomas Hugh Jones Barnsley Hospital NHSFT, Barnsley, UK; University of Sheffield, Sheffield, UK Testosterone deficiency is common in men with cardiovascular disease (CVD) and/or Type 2 Diabetes (T2D). Several studies have shown that testosterone replacement therapy (TRT) improves key cardiovascular risk factors such as insulin resistance, glycemic control, dyslipidaemia, obesity, hypertension. Low testosterone is also associated with non-alcoholic liver disease (NAFLD), a condition associated with insulin resistance and CVD.[br]This retrospective audit of 401 hypogonadal men from normal clinical practice (47.4% T2D, 34% CVD) reviewed the effects of long-term (mean 5.41 years; 1642 patient years) physiological TRT i.e. maintaining testosterone levels within the mid to upper normal range on BMI, waist circumference, blood pressure (BP), lipid profile, liver function, testosterone and estradiol levels, with measurements being taken at 3, 6 and 12 months and yearly thereafter. Data from the most recent visit represents the primary endpoint.[br]Testosterone was brought up from a baseline mean level of 7.16[plusmn]2.59nmol/l to 17.60[plusmn]9.82nmol/l at the primary endpoint and oestradiol increased by 28.96pmol/l(P[lt]0.005). Total cholesterol (-0.28mmol/l, p=0.004), triglycerides (-0.244mmol/l, p=0.040) and LDL (-0.19 mmol/l, p=0.033), HDL (-0.053mmol/l,p0.044) all showed a significant reduction at the primary endpoint. Liver transaminases ALT (-4.96iu/l,p=0.015) and AST (-3.34iu/l,p=0.017) were reduced. A reduction in waist circumference approached significance (p=0.059). No significant reduction was seen in weight, BMI, or blood pressure. In men with T2D HbA1c[gt]7.0% (n=151) a significant fall in HbA1c was observed at 3 (8.65 v 7.65%,p[lt]0.001 and at primary endpoint 6.74%). There were few changes required in glycemic drugs and only 2 subjects required the addition of insulin.[br]This audit supports a long-term improvement of lipid parameters although found as shown in clinical trials a small lowering of HDL-cholesterol. The decrease in liver transaminases is suggestive that testosterone improves hepatic steatosis. A clinical relevant reduction in HbA1c was observed in poorly controlled men with T2D. TRT has an overall beneficial effect on reducing cardiovascular risk factors in hypogonadal men.[br][br]Nothing to Disclose: DSM, JCB, DJW, VM, THJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2177 293 1994 MON-44 PO36-01 Monday 1662 2012


1659 ENDO12L_MON-45 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) A Preliminary Trial of Testosterone Replacement for Fatigue in Male Hypogonadal Patients with Advanced Cancer Diana Engineer, Gina Cardwell, Eduardo Bruera, J Lynn Palmer, Egidio del Fabbro, Jose Garcia Michael E Debakey Veterans Affairs Medical Center, Houston, TX; Baylor College of Medicine, Houston, TX; St Luke[apos]s Episcopal Hospital, Houston, TX; Baylor College of Medicine, Houston, TX; University of Texas MD Anderson Cancer Center, Houston, TX [bold]Background:[/bold] Patients with advanced cancer frequently identify fatigue as their primary impairment. Hypogonadism may be a significant, potentially treatable, contributor to fatigue in male patients with advanced cancer. Previous studies have shown a high prevalence of hypogonadism in this population.[br][bold]Objective:[/bold] The primary endpoint was to evaluate the effect of testosterone replacement therapy on fatigue in hypogonadal male patients with advanced cancer, measured by the Functional Assessment of Cancer Therapy-Fatigue subscale (FACIT-F) at day 29.[br][bold]Design:[/bold] Randomized, double-blinded placebo controlled study conducted at the MDVAMC and UT MD Anderson Cancer Center. Patients with advanced cancer, bioavailable testosterone (BT) level [lt]70ng/dL and ECOG [lt]3 were included in the study. Those with any contraindication for testosterone therapy, Hgb[lt]9, or other medical causes of fatigue (CHF, COPD, CKD) were excluded. Eligible subjects were randomized to testosterone treatment or placebo. Testosterone was administered IM using a weight-based dose titrated every 14 days to goal BT 70-270ng/dL.[br][bold]Results:[/bold] Of the 48 subjects that met our criteria, 19 were randomized to testosterone and 24 to placebo. While no statistical significance was noted in age, there was a significant difference in race (p 0.02). No statistically significant differences in FACIT-F subscale scores were found between arms using a one-sided t test, although differences were in the expected direction (-1.7[plusmn]12 for placebo, 3.5[plusmn]7.7 for testosterone, p=0.105). Similar results were found for the FACIT-F total score with a trend for testosterone to improve scores (-5.5[plusmn]19 for placebo, 3.9[plusmn]14 for testosterone, p=0.09). One subject died in the testosterone arm and this was considered unrelated to the treatment. Adverse events were similar between groups.[br][bold]Conclusion: [/bold]Testosterone administration in hypogonadal, male patients with advanced cancer may improve fatigue and overall quality of life. Larger interventional studies to determine the degree of benefit and safety in this population are warranted.[br][br]Sources of Research Support: Study funded by the American Cancer Society. We acknowledge the support of MDACC and the Dept of VA.[br][br]Disclosures: JG: Investigator, Aeterna Zentaris Inc. Nothing to Disclose: DE, GC, EB, JLP, EdF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 585 293 1995 MON-45 PO36-01 Monday 1663 2012


1660 ENDO12L_MON-46 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Quality of Life, Testosterone and Abdominal Obesity in 779 Young and 598 Aging Men. Effects of Randomized Testosterone Treatment on Quality of Life in 38 Aging Men with Relative Hypogonadism Dorte Glintborg, Torben Leo Nielsen, Louise Frederiksen, Kristian Wraae, Claire Gudex, Kim Brixen, Marianne Andersen Odense University Hospital, Odense, Denmark [bold]Introduction[/bold]: Quality of life (SF36) may be impaired in male hypogonadism.[br][bold]Aim: [/bold]To evaluate the association between quality of life, hypogonadism, and abdominal obesity.[br][bold]Design:[/bold] Two population-based cross-sectional studies in 779 young (20[ndash]29 years) and 598 ageing (60-74 years) men and a randomized, double-blinded, placebo-controlled study of six months testosterone therapy (gel) in 38 ageing men with low-normal bioavailable testosterone ([lt] 7.3 nmol/l) and waist circumference [gt] 94 cm were performed.[br][bold]Methods:[/bold] SF36, testosterone, and clinical evaluation. Lean body mass (LBM) and total fat mass were established by dual x-ray absorptiometry and visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) were measured by magnetic resonance imaging. In the intervention study, [Delta]-values were calculated as post treatment level minus pre-treatment level. Responders were defined as testosterone treated patients with [Delta]LBM [gt] 0 kg, n=14. Data were presented as median (interquartile range).[br][bold]Results:[/bold] In young men (n=779), low-normal bioavailable testosterone and increased waist circumference were associated with decreased SF36 physical domain scores.[br]Testosterone treatment was associated with increased LBM and decreased SAT. In responders (n=14), the MH-score significantly increased during testosterone treatment [84 (76-92) vs. 96 (88-100)], p=0.02. [Delta]LBM was positively associated with [Delta]VT (r= 0.57), [Delta]RE (r= 0.56), and [Delta]MCS (r= 0.47), (n=14, all p[lt]0.05). [Delta]SF36 outcomes were not associated with [Delta]testosterone. At the ENDO meeting additional data on SF36, testosterone levels, and body composition in elderly men will be presented.[br][bold]Conclusion:[/bold] SF36 physical domain scores were associated with low-normal bioavailable testosterone and increased waist circumference in young men. Increased quality of life during testosterone treatment was associated with improved body composition, whereas no associations were found with [Delta]testosterone levels in ageing men with low-normal bioavailable testosterone.[br][br]Sources of Research Support: Novo Nordisk Foundation, WADA, Antidoping Denmark.[br][br]Nothing to Disclose: DG, TLN, LF, KW, CG, KB, MA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 796 293 1996 MON-46 PO36-01 Monday 1664 2012


1661 ENDO12L_MON-47 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Long-Term Testosterone Treatment with Injectable Testosterone Undecanoate in Hypogonadal Men with Inflammatory Bowel Diseases (M. Crohn and Colitis ulcerosa) Farid Saad, Ahmad Haider, Winfried Kurtz, Mahmoud Nasser, Louis Gooren Bayer Pharma AG, Berlin, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates; Private Practice, Bremerhaven, Germany; Klinikum Bremerhaven, Bremerhaven, Germany; University of Aleppo, Aleppo, Syrian Arab Republic; VUMC Amsterdam, Amsterdam, Netherlands Objective: Testosterone (T) has anti-inflammatory effects. T treatment has been found to be beneficial in rheumatoid arthritis and chronic obstructive pulmonary disease. We had reported effects of two years of T treatment in a small group of hypogonadal men with Crohn[apos]s disease (Haider A et al., Horm Mol Biol Clin Invest 2010; 2(3): 287[ndash]292).[br]Methods and Design: 48 men with Crohn[apos]s disease and 2 men with Colitis ulcerosa with T [le] 12 nmol/L from 2 centers in Bremerhaven, Germany and Damascus, Syria received treatment with parenteral testosterone undecanoate on day 1, after 6 weeks and thereafter every 12 weeks for up to 60 months. 12 hypogonadal men of similar age with Crohn[apos]s disease who did not receive T served as a control group. The Crohn[apos]s Disease Activity Index (CADI) was assessed as an indicator of the severity of the disease every 3 months. In addition, highly sensitive C-reactive protein (hsCRP) and leukocyte count as markers of inflammatory activity were measured.[br]Results: T levels at baseline were 2.83 [plusmn] 0.34 ng/mL in the T group and 3.1 [plusmn] 0.11 in the control group. During treatment, T increased to 5.27 [plusmn] 0.46 and remained stable at 3.01 [plusmn] 0.1 in the control group. hsCRP (mg/dL) levels at baseline were 16.45 [plusmn] 9.97in the T-treated group vs 7.43 [plusmn] 0.92 in the control group. They decreased to 3.82 [plusmn] 2.46 after 60 months in the T-treated group and increased to 9.25 [plusmn] 1.61 in the control group. The CADI decreased in the T-treated group from xxx [plusmn] xxx to xxx [plusmn] xx (p=xxx). In the control group, the CADI decreased from 220.61 [plusmn] 49.1 to 72.78 [plusmn] 2.64 in the treated group and increased from 196.82 [plusmn] 7.17 to 213.18 [plusmn] 14.19 in the control group. Leukocyte count decreased from 13.17 [plusmn] 2.7 to 5.94 [plusmn] 0.73 in the treated group and remained unchanged in the control group (from 11.43 [plusmn] 1.34 to 11.08 [plusmn] 1.46).[br]Conclusion: Normalization of T in hypogonadal men with Crohn[apos]s disease led to improvements of the CADI, hsCRP and a reduction of leukocytes.The mechanism of this improvement may be through anti-inflammatory and immunosuppressive effects of testosterone, reducing chronic inflammation of the intestinal wall in CD.[br][br]Haider A et al., Horm Mol Biol Clin Invest 2010; 2(3): 287[ndash]292.[br][br]Sources of Research Support: Data entry and statistical analysis were supported by Bayer Pharma AG.[br][br]Disclosures: FS: Employee, Bayer Schering Pharma. AH: Principal Investigator, Bayer Schering Pharma. LG: Speaker, Bayer Schering Pharma. Nothing to Disclose: WK, MN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 492 293 1997 MON-47 PO36-01 Monday 1665 2012


1662 ENDO12L_MON-48 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Treatment of Patients with a Late Diagnosis of Klinefelter Syndrome with Testosterone Undecanoate for a Duration of up to 5 Years Farid Saad, Ahmad Haider, Gheorghe Doros, Louis Gooren Bayer Pharma AG, Berlin, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates; Private Practice, Bremerhaven, Germany; School of Public Health, Boston University, Boston, MA; VUMC Amsterdam, Amsterdam, Netherlands Introduction: Males suffering from Klinefelter[apos]s Syndrome (KS) experience an increased hospitalization rate from a variety of disorders, some caused by hypogonadism, and some linked to the syndrome [italic]per se[/italic], others not readily explained, maybe socioeconomic. The degree of hypogonadism is variable with the vast majority below the normal range. Mortality in KS is significantly increased, with excess deaths due to diabetes, cardiovascular, respiratory, and gastrointestinal diseases. KS is markedly underdiagnosed with less than 10% of cases identified by puberty and less than 20% ever diagnosed during life.[br]In the present study we describe a cohort of men suffering from KS, diagnosed at advanced age. The majority of the patients had been referred by an orthopedic surgeon after a diagnosis of osteoporosis to be checked for hypogonadism.[br]Methods: Open-label, single-center, cumulative, prospective registry study of 22 middle-aged men with testosterone levels between 2.1 and 3.5 ng/mL (mean: 3.08 [plusmn] 0.46). 21 men were studied for at least 2 years, 18 for 3 years, 11 for 4 and 8 for at least 5 years. They received parenteral testosterone undecanoate 1000 mg/12 weeks after an initial interval of 6 weeks.[br]Results: After 5 years the following changes were observed: Testosterone (ng/mL) increased from 3.08 [plusmn] 0.46 to 4.86 [plusmn] 0.47 ng/mL (p[lt]0.0001). Body weight (kg) decreased from 105.05 [plusmn] 11.18 (minimum: 70, maximum: 139) to 90.88 [plusmn] 9.76 (p[lt]0.0001 vs baseline). Waist circumference (cm) declined from 104.32 [plusmn] 6.39 to 96.50 [plusmn] 6.63 (p[lt]0.0001 vs baseline). The mean T-score increased from -2.91 [plusmn] 0.31 (min -3.90, max -2.60) to -1.50 [plusmn] 0.28 (p[lt]0.0001 vs baseline). Glucose levels remained unchanged with mean levels below 100 mg/dL at all time-points. Total cholesterol decreased from 240.95 [plusmn] 18.89 to 177.75 [plusmn] 16.69 (p[lt]0.0001), LDL from 124.09 [plusmn] 46.4 to 64.13 [plusmn] 18.13 (p[lt]0.0001) and triglycerides from 235.09 [plusmn] 22.07 to 180.75 [plusmn] 7.59 (p[lt]0.0001). HDL did not change significantly.[br]Conclusions: Raising serum testosterone to normal in patients with a late diagnosis of Klinefelter[apos]s syndrome improved body composition including bone mineral density. Other features of the metabolic syndrome were also improved.[br][br]Sources of Research Support: Data entry and statistical analysis were supported by Bayer Pharma AG.[br][br]Disclosures: FS: Employee, Bayer Schering Pharma. AH: Principal Investigator, Bayer Schering Pharma. GD: Coinvestigator, Bayer Schering Pharma. LG: Speaker, Bayer Schering Pharma. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1070 293 1998 MON-48 PO36-01 Monday 1666 2012


1663 ENDO12L_MON-49 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Testosterone Treatment with Injectable Testosterone Undecanoate Sustainably Improves Erectile Function, Urinary Function and Quality of Life in Elderly Hypogonadal Patients Farid Saad, Ahmad Haider, Gheorghe Doros, Louis Gooren Bayer Pharma AG, Berlin, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates; Private Practice, Bremerhaven, Germany; School of Public Health, Boston University, Boston, MA; VUMC Amsterdam, Amsterdam, Netherlands [bold]Objectives:[/bold] Hypogonadism is often associated with erectile dysfunction (ED) and lower urinary tract symptoms (LUTS) and impaired Quality of Life. We studied long-term effects of testosterone treatment in elderly hypogonadal men treated with parenteral testosterone undecanoate.[br][bold]Methods:[/bold] A cumulative study of 255 men (mean age: 60.6 [plusmn] 8.0 years) with testosterone levels [le] 3.50 ng/mL were treated with injectable testosterone undecanoate for up to 60 months. Injections were administered with an initial 6-week interval (loading dose) followed by 12-week intervals.[br][bold]Results:[/bold] The International Index of Erectile Function (IIEF) increased from 21.13 [plusmn] 4.63 at baseline to 24.83 [plusmn] 3.8 after 60 months, most pronounced over the first 24 months but still slowly progressive thereafter. The International Prostate Symptoms Score (IPSS) improved from 6.73 [plusmn] 4.21 to 2.83 [plusmn] 1.25 (p[lt]0.0001 vs baseline with significant changes over the previous year up to 48 months). As an objective measurement, residual bladder volume decreased from 46.61 [plusmn] 22.74 mL to 19.74 [plusmn] 6.25 mL (p[lt]0.0001 vs baseline with significant changes over the previous year up to 48 months). Quality of life was assessed by the Aging Males[apos] Symptoms score (AMS). AMS improved from 55.01 [plusmn] 10.2 to 17.35 [plusmn] 0.55 (p[lt]0.0001 vs baseline) reaching a plateau after 24 months.[br]Inflammation seems to play a role in both erectile and urinary function. As measures of inflammation, highly sensitive C-reactive protein (hsCRP) and leukocyte count were assessed. hsCRP decreased from 6.29 [plusmn] 7.96 mg/L to 1.03 [plusmn] 1.87 (p[lt]0.0001 vs baseline) with a plateau after 36 months. Leukocyte count decreased from 8.06 [plusmn] 2.98 x10[sup]9[/sup]/L to 5.74 [plusmn] 0.81 (p[lt]0.0001 vs baseline).[br][bold]Conclusions:[/bold] Sustainable and progressive improvement of IIEF and IPSS was remarkable in conjunction with improvement of body composition, lipid metabolism, inflammatory markers and blood pressure. We assume that ischemia, impairment of nitric oxide (NO) production and inflammation play a role in the etiology of ED and LUTS which often occur in a parallel pattern. Testosterone treatment in hypogonadal men may reduce inflammation and may have direct effects on NO mechanisms and reduction of ischemia resulting in improvement of erectile and voiding capacities.[br][br]Sources of Research Support: Data entry and statistical analysis were supported by Bayer Pharma AG.[br][br]Disclosures: FS: Employee, Bayer Schering Pharma. AH: Principal Investigator, Bayer Schering Pharma. GD: Coinvestigator, Bayer Schering Pharma. LG: Speaker, Bayer Schering Pharma. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1074 293 1999 MON-49 PO36-01 Monday 1667 2012


1664 ENDO12L_MON-50 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Side-Effect Profile of Long-Term Treatment of Elderly Hypogonadal Men with Testosterone Undecanoate Farid Saad, Ahmad Haider, Gheorghe Doros, Louis Gooren Bayer Pharma AG, Berlin, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates; Private Practice, Bremerhaven, Germany; Public School of Health, Boston University, Boston, MA; VUMC Amsterdam, Amsterdam, Netherlands [bold]Introduction:[/bold] Testosterone therapy for hypogonadal men has been used for decades. However, there are still concerns regarding the safety of this treatment, particularly in elderly men.[br][bold]Methods:[/bold] Prospective registry study of 255 men (mean age 60.6 [plusmn] 8.0 years), with testosterone levels between [le] 3.5 ng/ml. They received parenteral testosterone undecanoate 1000 mg at day 1, week 6 and every 12 weeks thereafter for up to 66 months.[br][bold]Results:[/bold] After 60 months the following changes were observed:[br]Erythropoiesis: haemoglobin increased from14.44 [plusmn] 0.72 to 14.99 [plusmn] 0.45 g/dl (p[lt]0.0001 vs baseline). Haematocrit increased from 43.22 [plusmn] 2.84 to 48.78 [plusmn] 1.7% (p[lt]0.0001 vs baseline). Four patients had haematocrit levels [gt] 52% which resolved without intervention.[br]Prostate: PSA increased from 1.77 [plusmn] 0.96 to 1.82 [plusmn] 0.96 ng/ml (p[lt]0.0001 vs baseline) with a plateau after 24 months. Prostate volume increased from 28.51 [plusmn] 11.2 to 30.23 [plusmn] 12.4 ml (p[lt]0.0001 vs baseline). 3/255 patients were diagnosed with prostate cancer following elevated PSA ([lt] 4 ng/mL) at 18 weeks of treatment. Tumour grade was T2 in all three and Gleason score 3+3 in two and 3+2 in one patient, resp. They all underwent radical prostatectomy. The proportion was 1.18% with an incidence of 30.334 per 10.000 patient years. For comparison: in the PLCO trial with a 7-year follow-up, the proportion of prostate cancer was 7.35% with an incidence of 116 per 10.000 patient years [1]. [ndash] The International Prostate Symptom score (IPSS) improved from 6.73 [plusmn] 4.21 to 2.83 [plusmn] 1.25 (p[lt]0.0001).[br]Liver enzymes: aspartate transaminase (AST) decreased from 43.05 [plusmn] 17.29 to 20.16 [plusmn] 3.21 U/L (p[lt]0.0001 vs baseline) reaching a plateau after 24 months, alanine transaminase (ALT) from 43.89 [plusmn] 18.11 to 20.54 [plusmn] 3.92 U/L (p[lt]0.0001 vs baseline) with a plateau after 36 months.[br][bold]Conclusions:[/bold] The incidence of 3/255 patients with prostate cancer does not suggest an increased risk of prostate cancer in elderly men on long-term testosterone treatment. Long-term treatment with testosterone undecanoate with monitoring according to the guidelines is acceptably safe.[br][br](1) Andriole G et al., New Engl J Med 2009; 360(13): 1310-1319.[br][br]Sources of Research Support: Data entry and statistical analysis were supported by Bayer Pharma AG.[br][br]Disclosures: FS: Employee, Bayer Schering Pharma. AH: Principal Investigator, Bayer Schering Pharma. GD: Coinvestigator, Bayer Schering Pharma. LG: Speaker, Bayer Schering Pharma. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1071 293 2000 MON-50 PO36-01 Monday 1668 2012


1665 ENDO12L_MON-51 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Long-Term Testosterone Treatment in Hypogonadal Men Improves All Components of the Metabolic Syndrome Aksam Yassin, Youssef El Douaihy, Ridwan Shabsigh, Aiman Yassin, Farid Saad Segeberger Kliniken, Norderstedt, Germany; Dresden International University, Dresden, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates; Maimonides Medical Center, Brooklyn, NY; Klinikum Braunschweig, Braunschweig, Germany; Bayer Pharma AG, Berlin, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates Introduction: Metabolic syndrome as well as all its individual components are associated with testosterone deficiency. We hypothesize that long-term testosterone therapy improves the metabolic syndrome.[br]Materials and Methods: Since November 2004, 261 patients diagnosed with LOH in our center in Germany were entered into our registry study for long term testosterone replacement therapy. Men with a total testosterone of [le]3.5 ng/mL and symptoms of erectile dysfunction (IIEF-5[lt] 21) met the inclusion criteria. Long acting testosterone undecanoate 1000 mg injections were administered at day 1, after 6 weeks and every 3 months thereafter. The mean follow up time was 4.25 years. Demographic data were collected at the baseline. Hormones, lipids, glucose and blood pressure were measured at every visit or every other visit. We used the International Diabetes Federation definition to define whether patients had the metabolic syndrome (MetS). MetS was considered in patients with central obesity and any two of the following: raised triglycerides, reduced HDL cholesterol, raised blood pressure, and raised fasting plasma glucose. Mean fasting glucose, triglycerides, and HDL were calculated at every T and linear regression was used to study their variation over time. Total testosterone and prevalence (%) of the MetS were plotted against time.[br]Results: we noticed a significant drop in the prevalence of the MetS from 56% to 30% after 57 months. There was a statistically significant drop of triglycerides, glucose, and the mean arterial pressure (p=0.00; [beta]=-0.77, p=0.00; [beta]=-0.67, and p=0.00; [beta]=-0.78respectively) and an increase in HDL (p=0.05; [beta]=0.53). In addition, we noticed a significant drop of 11 cm in the mean waist circumference.[br]Conclusion: All components of the MetS improved significantly and over the full observation period upon normalization of testosterone levels.[br][br]Sources of Research Support: Data entry was supported by Bayer Pharma AG.[br][br]Disclosures: AY: Speaker, Bayer Schering Pharma. RS: Speaker, Lilly USA, LLC. FS: Employee, Bayer Schering Pharma. Nothing to Disclose: YED, AY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 460 293 2001 MON-51 PO36-01 Monday 1669 2012


1666 ENDO12L_MON-52 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Sustained Improvement of Features of the Metabolic Syndrome upon Normalization of Serum Testosterone in Hypogonadal Men: Follow-Up up to 5 Years Farid Saad, Ahmad Haider, Gheorghe Doros, Louis Gooren Bayer Pharma AG, Berlin, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates; Private Practice, Bremerhaven, Germany; School of Public Health, Boston University, Boston, MA; VUMC Amsterdam, Amsterdam, Netherlands Objectives: Hypogonadal men tend to increase body weight, fat mass and develop features of the metabolic syndrome. Long-term effects of normalization of testosterone in hypogonadal men on weight, waist circumference and lipid metabolism, liver functions upon treatment with parenteral testosterone undecanoate were studied.[br]Methods: A cumulative registry study of 255 men (mean age: 60.6 [plusmn] 8.0 years), with testosterone levels between 0.14 [ndash]3.51 ng/mL with late onset hypogonadism (LOH).[br]Results: A remarkable progressive and sustained decline of body weight and waist circumference over 5 years was observed. Fasting glucose decreased from 103.38 [plusmn] 14.44 mg/dL to 97.54 [plusmn] 2.34 (p[lt]0.0001). The proportion of patients who had glucose levels [ge] 100 mg/dL decreased from 45% at baseline to 16% at 60 months. HbA[sub]1c[/sub] was measured in 125 patients and decreased from 6.94 [plusmn] 1.55% to 6.01 [plusmn] 1.41%. Total cholesterol decreased from 281.58 [plusmn] 39.8 mg/dL to 188.12 [plusmn] 11.31 (p[lt]0.0001), LDL from 163.79 [plusmn] 41.44 mg/dL to 109.84 [plusmn] 35.41 (p[lt]0.0001), triglycerides from 276.16 [plusmn] 51.32 mg/dL to 189.78 [plusmn] 11.33 (p[lt]0.0001). HDL was stable over the first 2 years (62 mg/dL at baseline and 63.26 at 24 months) and then declined to 52.45 at 60 months (p[lt]0.0001 vs baseline). Mean systolic blood pressure declined from 153.55 [plusmn] 17.6 mm Hg to 137.74 [plusmn] 10.92 (p[lt]0.0001) and diastolic blood pressure from 93.49 [plusmn] 11.21 mm Hg to 79.61 [plusmn] 7.35 at 60 months (p[lt]0.0001). At baseline, 91% of men had a systolic blood pressure of [ge] 130 mm Hg which declined to 80% after 60 months. At baseline, 75% of men had a diastolic blood pressure of [ge] 85 mm Hg declining to 22% at 5 years.[br]Conclusions: Normalization of serum testosterone leads to a sustained improvement of all components of the metabolic syndrome.[br][br]Sources of Research Support: Data entry and statistical analyses were supported by Bayer Pharma AG.[br][br]Disclosures: FS: Employee, Bayer Schering Pharma. AH: Principal Investigator, Bayer Schering Pharma. GD: Coinvestigator, Bayer Schering Pharma. LG: Speaker, Bayer Schering Pharma. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 452 293 2002 MON-52 PO36-01 Monday 1670 2012


1667 ENDO12L_MON-53 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) [ldquo]Late Onset Hypogonadism[rdquo] Is Not an Isolated Condition [mdash] Comorbidities in Elderly Hypogonadal Men Presenting or Referred to Urological Institutions in Germany Ahmad Haider, Aksam Yassin, Louis Gooren, Farid Saad Private Practice, Bremerhaven, Germany; Segeberger Kliniken, Norderstedt, Germany; Dresden International University, Dresden, Germany; Gulf Medical University School of Medicine, Ajman, United Arab Emirates; VUMC Amsterdam, Amsterdam, Netherlands; Bayer Pharma AG, Berlin, Germany; Gulf Medical University School, Ajman, United Arab Emirates Introduction: Serum testosterone declines with aging, not primarily determined by calendar age per se but rather by factors impairing the health of aging men, such as obesity, metabolic syndrome, diabetes mellitus and other diseases. We determined concurrent diseases in two cohorts of mainly elderly men with so-called late onset hypogonadism (LOH).[br]Methods: In two separate cumulative registry studies following identical protocols, two cohorts of 516 mainly elderly men were analyzed for concurrent diseases. Cohort A (Dr. Haider, Bremerhaven, Germany) consisted of 255 men, cohort B (Dr. Yassin, Norderstedt, Germany) of 261 men. These men had either sought urological consultation or had been referred by other disciplines because of suspected hypogonadism. All men received treatment with injections of long-acting testosterone undecanoate.[br]Results: The following comorbidities were encountered:[br]Cardiology: hypertension: A: 40%, B: 45%; coronary artery disease: A: 16%, B: 13%; condition post myocardial infarction: A 15%, B: [lt]1%.[br]Internal Medicine: Diabetes mellitus: A: 31%, B: 26%; dyslipidemia: A: 18%, B: 33%.[br]Gastroenterology: inflammatory bowel disease: A: 16%, B: [lt]1%.[br]Urology: chronic prostatitis: A: 38%, B: 11%.[br]Dermatology: psoriasis: A: 5%, B: [lt]1%.[br]Endocrinology: Klinefelter[apos]s syndrome: A: 9%, B: 2%. In addition, there were a total of 14 patients with a history of maldescensus testis and 19 patients with a history of unilateral or bilateral orchiectomy following testicular cancer.[br]Orthopedics: osteoporosis: A: 14%, B: 6%.[br]Conclusions: 1) The majority of middle-aged to elderly patients with hypogonadism have one or more comorbidities. For adequate treatment, hypogonadal men should be examined for concurrent diseases. Testosterone administration may be a significant element in their treatment. 2) With progression of their age elderly men will suffer increasingly from ailments and hypogonadism may be an element, so far not often diagnosed. Testosterone treatment may contribute to a better quality treatment. 3) 60/516 men had conditions which cannot be categorized as [ldquo]LOH[rdquo]. Klinefelter[apos]s syndrome may still have been undiagnosed, and a history of maldescensus testis may be unknown. The term [ldquo]LOH[rdquo] should be used with caution, and the general term [ldquo]hypogonadism[rdquo] may be preferable.[br][br]Sources of Research Support: Data entry was supported by Bayer Pharma AG.[br][br]Disclosures: AH: Principal Investigator, Bayer Schering Pharma. AY: Speaker, Bayer Schering Pharma. LG: Speaker, Bayer Schering Pharma. FS: Employee, Bayer Schering Pharma. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1082 293 2003 MON-53 PO36-01 Monday 1671 2012


1668 ENDO12L_MON-54 POSTER SESSION: Male Reproductive Endocrinology (1:30 PM-3:30 PM) Hormone Replacement Therapy in Young Men with Hypogonadism: Quality of Life According IPSS, AMS and IIEF Scores Marta Snajderova, Karim El Balouly, Vladimir Sobotka, Daniela Zemkova, Michael Urban, Jiri Heracek Charles University, 2nd Faculty of Medicine and University Hospital Motol, Prague, Czech Republic; Charles University, 3rd Faculty of Medicine and University Hospital Kralovske Vinohrady, Prague, Czech Republic [bold]Objectives: [/bold]Male hypogonadism has significant effects on the fertility, sexual function and general health of patients. Testosterone (T) replacement is the most effective treatment for sexual dysfunction. We evaluated quality of life by questionnaires, IPSS (International Prostate Symptom Score), the AMS (Aging Male Score) and IIEF (International Index of Erectile Function) during continuous long-term hormone replacement therapy (HRT) by testosterone.[br][bold]Methods: [/bold]During the period 2005-2010, we investigated 23 patients (aged 22-36 years) with hyper- or hypogonadotropic hypogonadism treated by Sustanon[reg] i.m. (Organon) every 3 weeks or Nebido[reg] i.m. (Bayer Schering Pharma AG) every 3 months. The mean follow-up period was 3.8 years (2 months - 5 years). Quality of life by questionnaires IPSS, IIEF and AMS was evaluated before the onset of HRT and at the end of follow-up.[br][bold]Results: [/bold]The average IIEF score of the set improved by 15%, markedly for erectile and orgasmic scores (pretreatment/treatment 13/19; and 3/6). The average value of IPSS did not change (4/5) during the follow-up period. AMS declined by 42% (36/21). In the majority of subjects no side effects of HRT were present. Only one patient (4%) experienced depression requiring hospitalization, and in another two (9%) men strong sweating and flushing occurred.[br][bold]Conclusions: [/bold]The best improvement during long-term male HRT was found in AMS. Erectile and orgasmic scores improvement was detected as well. This work presents the results of long-term comprehensive care for patients with impaired sexual development. Long-term monitoring in interdisciplinary collaboration (andrologist [amp] endocrinologist) may help to clarify the impact of T replacement therapy on quality of life and on development of androgen-dependent organs.[br][br]Sources of Research Support: The project IGA MZ CR NS9983.[br][br]Nothing to Disclose: MS, KEB, VS, DZ, MU, JH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 417 293 2004 MON-54 PO36-01 Monday 1672 2012


1669 ENDO12L_MON-55 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) Long-Term Follow-Up of Hormonal Gonadal Function of 46,XX Patients with Disorder of Sex Development (DSD) Due to Abnormalities of Gonadal Determination Patricia Sales Marques da Cruz, Rosana Barbosa Silva, Elaine Maria Frade Costa, Mirian Yumi Nishi, Berenice Bilharinho Mendonca, Sorahia Domenice Hospital das Cl[iacute]nicas da Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil 46,XX DSD due to abnormalities of gonadal determination (AGD) is a rare condition. This particular disorder allows us to analyze the impact of the lacking Y chromosome genes on male gonadal tissue in 46,XX DSD patients with AGD. [bold]Aim-[/bold] To describe the follow-up of the gonadal function of 46,XX DSD patients due to AGD ([italic]SRY[/italic]+ or [italic]SRY[/italic] negative). [bold]Subjects- [/bold]Nineteen 46,XX ovotesticular DSD (OTDSD) and 9 46,XX testicular DSD (TDSD) patients were studied. Complete bilateral gonadectomy was done in 9 OTDSD. Conservative ovarian tissue surgery was done in 2 OTDSD girls and testicular tissue was preserved in 8 OTDSD male patients. All the 15 OTDSD patients studied and 3 ambiguous TDSD patients was [italic]SRY [/italic]negative. Five TDSD patients with male genitalia were [italic]SRY[/italic]+. [bold]Results-[/bold] Puberty started spontaneously at age of 11 to 14 yrs in 14 OTDSD (5 of them after conservative gonadal surgery) and 7 TDSD. Four patients (2 OTDSD and 2 TDSD) were at pre pubertal age. The 2 girls had spontaneous menarche; one girl had a premature ovarian failure at 18 yrs old and the other has LH and FSH normal levels and menstrual cycle at 20 yrs old. Ten males (3 OTDSD and 7 TDSD), who developed spontaneous puberty need testosterone substitution. [bold]Discussion-[/bold] The presence of [italic]SRY[/italic] in 46,XX TDSD patients was critical in determining testicular hormones secretion and genital virilization during fetal life. However, independently of the genetic mechanism that determine the testicular differentiation in the 46,XX fetal gonad, this testicular tissue undergoes a degenerative process and a progressive failure of hormonal secretion. Models of [italic]Sry[/italic]+ XX male mice demonstrated that their testes were defective in supporting the later phases of spermatogenesis and probably insufficient to induce adequate Sertoli cell function. These studies suggest that Y chromosomal genes may be crucial for spermatogenesis and for somatic cell function. [bold]Conclusion-[/bold] In this cohort of 46,XX DSD patients, the fetal testicular hormonal function of all [italic]SRY+ [/italic]TDSD patients supported the development of the internal and external male genitalia. However, a progressive loss of testicular hormonal secretion was observed in all [italic]SRY+[/italic] and [italic]SRY [/italic]negative TDSD patients, and in the half of 46,XX [italic]SRY [/italic]negative OTDSD patients. These findings reinforce the hypothesis that Y chromosomes genes may be important in the maintenance of the testicular cells and not only in the spermatogenesis.[br][br]Nothing to Disclose: PSMdC, RBS, EMFC, MYN, BBM, SD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 972 294 2005 MON-55 PO37-01 Monday 1673 2012


1670 ENDO12L_MON-56 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) Individualizing Care for Patients with Gonadal Dysgenesis: A Case Series Jessica Crawford, Lynda Jacks, Lefkothea Karaviti, David Roth, Jennifer Bercaw-Pratt, Xiomara Santos, Jennifer Dietrich Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX Introduction: Gonadal dysgenesis (GD) occurs in 1/4500 infants, and may present with ambiguous genitalia. We sought to explore the anatomic findings associated with GD.[br]Background: The diagnosis, management and treatment for patients with disorders of sexual differentiation is based on genetics, physical exam findings, laboratory evaluation and family discussion. There are no clear guidelines regarding the predictability of anatomic findings associated with specific disorders of sexual development (DSD).[br]Objective: To review patients with GD followed in a Gender Medicine Clinic (GMC) to determine if any consistencies in physical exam findings, imaging studies or surgery may aide in diagnosis and treatment. Our hypothesis was that no anatomic findings are universally associated with a diagnosis of GD.[br]Clinical Course: A retrospective chart review of a GMC database was performed between 2008-2011, and data regarding anatomic findings was collected. Descriptive statistics were performed.[br]Results: Chart review yielded 37 patients with a diagnosis of GD. Of these, 35.1% (N=13) were assigned male gender and 64.9% (N=24) female gender. One individual in the male category had an unclear diagnosis of vanishing testis, and was excluded. Two individuals in the female category were excluded for possible diagnosis of partial androgen insensitivity or 46XX without other findings except delayed puberty and premature ovarian insufficiency. All 12 male patients included in the anatomic evaluation carried a diagnosis of mixed GD (46XY, 46XO/46XY, 46XX/47XXY). Of these, 30% presented with ovotestis, 30% with M[uuml]llerian remnants, 58.3% with abnormal penile length, 60% with hypospadias, chordee or micropenis and 25% with an abnormal appearing scrotum. In the female group, 0.9% had Swyer syndrome, 68.1% had Turner syndrome and 2.3% had mixed GD. Both patients with Swyer had a uterus, normal external genitalia and intrabdominal testes. All Turner[apos]s syndrome patients had normal appearing internal and external genitalia. Of patients with mixed GD (46XY, 46XX or 45XO/46XY), 40% had no evidence of M[uuml]llerian structures, 1 individual had a persistent urogenital sinus, 1 had a uterine remnant, and 1 had a vaginal remnant.[br]Conclusions: There were no anatomic findings consistently predictive of a specific diagnosis in the spectrum of GD.[br]Future Endeavors: We plan to expand this database, as ultimately, a larger cohort will be needed to make specific recommendations.[br][br]Nothing to Disclose: JC, LJ, LK, DR, JB-P, XS, JD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 351 294 2006 MON-56 PO37-01 Monday 1674 2012


1671 ENDO12L_MON-57 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) Long-Term Cross-Sex Hormone Therapy Changes Body Composition and Glucose Metabolism in Trans Women Katrien Wierckx, Youri Taes, Eva Van Caenegem, Greet Roef, Ruige Johannes, Jean-Marc Kaufman, Guy T[apos]Sjoen University Hospital Ghent, Ghent, Belgium [bold]Introduction: [/bold]Sex reassignment surgery and hormone replacement therapy drastically alters the hormonal status of male-to-female transsexual persons (trans women). As sex-steroids play an important role in body composition and glucose metabolism, long-term effects of anti-androgen and oestrogen treatment on glucose metabolism are important to assess in these individuals.[br][bold]Design: [/bold]Cross-sectional study, with 27 trans women before therapy and 31 subjects at least 1 year after sex reassignment surgery and on average 8.5 years on cross-sex hormone therapy, compared to 54 and 62 age-, weight- and height-matched control males, respectively.[br][bold]Methods: [/bold]Body composition (total and regional fat distribution) was determined using dual-energy X-ray absorptiometry (DXA) and peripheral quantitative computed tomography (pQCT). Commercial immunoassays were used to measure serum concentrations of insulin, leptin, estradiol, LH, testosterone and SHBG.[br][bold]Main results: [/bold]Baseline trans women were not different from control men with regard to serum sex steroid status, body composition and indices of glucose metabolism (fasting glucose, fasting insulin, HOMA-IR and QUICKI). However, trans women with on average 8.5 years of cross-sex hormone treatment presented lower total lean body mass, higher total body fat mass and subcutaneous fat area (proximal forearm and calf) compared to controls (all [italic]p[/italic][lt]0.001). Additionally, a higher limbfat/trunkfat ratio was observed in trans women ([italic]p[/italic][lt]0.001) as limb fat mass (mainly SC fat) was markedly higher in trans women. Trans women had lower serum fasting glucose but higher leptin (all [italic]p[/italic][lt]0.001). Also, fasting insulin, HOMA-IR and QUICKI were lower than in control men indicating higher insulin sensitivity in trans women, after adjustment for leptine ([italic]p[/italic]=0.036,[italic] p[/italic]=0.039 and [italic]p[/italic]=0.009, respectively).[br][bold]Conclusions: [/bold]Long term cross sex hormone administration in trans women results in different body composition with more fat mass, although insulin sensitivity compared to age-, height- and weight-matched male controls is higher after adjustment for fat mass.[br][br]Nothing to Disclose: KW, YT, EVC, GR, RJ, J-MK, GT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 451 294 2007 MON-57 PO37-01 Monday 1675 2012


1672 ENDO12L_MON-58 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) Low Bone Mass Is Prevalent in Transsexual Women before the Start of Cross-Sex Hormonal Therapy and Gonadectomy Eva Van Caenegem, Youri Taes, Katrien Wierckx, Sara Vandewalle, Greet Roef, Jean-Marc Kaufman, Thomas Schreiner, Guy T[apos]Sjoen Ghent University Hospital, Ghent, Belgium; Ghent University Hospital, Ghent, Belgium; Oslo University Hospital, Oslo, Norway Context:[br]Cross-sex hormonal therapy and sex reassignment surgery (SRS, including gonadectomy) in transsexual persons will to lead to an altered pattern of fat distribution and changes in bone mass and size.[br]Objective:[br]To examine body composition, bone mass and geometry in male-to-female transsexual persons (transsexual women) before cross-gender sex steroid exposure and SRS in comparison to age- and height-matched control men.[br]Design:[br]A prospective intervention study with 42 transsexual women (median age 31 years) with age-and height-matched control men. Grip strength, using a hand dynamometer, physical activity, areal bone mineral density (aBMD), using bone densitometry (DXA), bone geometry and volumetric bone mineral density (vBMD), using peripheral quantitative computed tomography, were measured, before the start of cross-sex hormonal therapy and SRS.[br]Results:[br]No differences were observed in weight, height, BMI, smoking and physical activity scores or fracture prevalence between transsexual women and controls. Body composition was also similar without significant differences in lean and fat mass, muscle mass and strength and waist and hip circumference. Prevalence of osteoporosis was higher in transsexual women (21%) compared to control men (3.4%) (p[lt] 0.05). They had a lower aBMD at the lumbar spine, hip and femoral neck (all p[le]0.01) and lower trabecular and cortical vBMD at the radius and tibia (all p[lt]0.05) compared to control men. A significantly thinner radial and tibial cortex (both p[le]0.016), possibly due to a smaller periosteal and larger endosteal circumference (n.s.), was also found in transsexual women compared to control men.[br]Positive associations were found between physical activity, in particular participation in sports, and aBMD at the lumbar spine, total hip and femoral neck in transsexual women (all p[lt]0.05), but not in control men.[br]Conclusions:[br]Transsexual women before the start of hormonal therapy appear to have a lower bone mass compared to control men. These baseline differences in bone mass might be related to physical activity. These findings have an impact on the interpretation of bone mineral density in the clinical management of transsexual women.[br][br]Nothing to Disclose: EVC, YT, KW, SV, GR, J-MK, TS, GT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 755 294 2008 MON-58 PO37-01 Monday 1676 2012


1673 ENDO12L_MON-59 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) High Frequency of Non-Prescription Hormone Treatment in Mexican Transgender Population with High Risk for HIV: Results from Patients in the First Free Care Clinic in Latin America Claudia Ramirez Renteria, Hamid Vega, Gabriela Hernandez, Victor Rodriguez Clinica Especializada Condesa, Health Department of Mexico City, Mexico City, Mexico; Clinica Especializada Condesa, Health Department of Mexico City, Mexico City, Mexico [bold]Background[/bold]: Health care for transgender people is still controversial in Latin America. The cost of medical care is prohibitive for most people in Mexico, specialized treatment is rarely requested and hormones are available without prescription. Low income patients are susceptible to dangerous behaviors in order to afford treatment, increasing HIV and STD risk. In 2009 our facility became the first totally free, government-funded, multidisciplinary transgender clinic in Latin America, providing medical, endocrinological and psychiatric care.[br][bold]Methods: [/bold]Clinical histories from July 2009 to January 2012 were reviewed for complete demographic information, including HIV status. Non parametric statistical tests were performed for comparisons, p values [lt] 0.05 were considered significant.[br][bold]Results[/bold]: At initial evaluation of 215 cases, 89.3% were male to female (MtF) and 137 (63.7%) had used hormones without prescription. The patients that used hormones before had been doing so intermittently for a median of 3 years (Interquartile ranges [IR] of 6 months to 11 years), they were significantly older (33 vs 24.5 years, p[lt]0.001), had higher frequency of HIV diagnosis (31.4% vs 9.1, p=0.01) and had a lesser scholarity -of 9 years or less (34.3% vs 19.7%, p=0.03)-. On the other hand, lipid profile was better on these patients; they had lower total cholesterol (169 vs 196 ng/dL, p=0.02) and LDL (91 vs 110 ng/dL, p=0.03) with no significant differences in body max index, triglycerides or HDL.[br][bold]Conclusions:[/bold] Our current data suggests that risk factors for HIV infection such as education and psychiatric support have significant differences between groups, also these risk factors are associated with the higher frequency use of non-prescription hormones. This cohort[apos]s characteristics are different from other international reports, mainly due to the availability of diverse over-the-counter hormonal treatments. However, it represents a large, high risk proportion of the transgender population in Mexico. Further investment, education and legislation in these areas is definitely needed in order to change the risk profile for the Mexican transgender population.[br][br]Nothing to Disclose: CRR, HV, GH, VR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 251 294 2009 MON-59 PO37-01 Monday 1677 2012


1674 ENDO12L_MON-60 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) Self-Reported Female Genital Sensitivity and Anatomy in a Dutch Speaking Population: Implications for Feminizing Surgery Guy Bronselaer, Nina Callens, Justine M Schober, Heino FL Meyer-Bahlburg, Griet De Cuypere, Guy T[apos]Sjoen, Martine Cools, Piet Hoebeke University Hospital Ghent and Ghent University, Ghent, Belgium; University Hospital Ghent and Ghent University, Ghent, Belgium; Hamot Medical Centre, Erie, PA; College of Physicians and Surgeons of Colombia University, New York, NY; University Hospital Ghent and Ghent University, Ghent, Belgium; University Hospital Ghent and Ghent University, Ghent, Belgium [italic][underline]Background:[/underline][/italic] Although (cosmetic) genitoplasty procedures for feminizing genitalia in women with a disorder of sex development (DSD) have much improved lately, little is known about self-perceived genital anatomy and long-term sensitivity. Moreover, the impact of the underlying DSD condition in determining psychosexual outcome cannot be underestimated. Therefore, self-reported judgment of anatomy and sensation and comparison between patients with or without a history of genital surgery are of utmost importance, but no normative data, based on a large sample, exist to date. [italic][underline]Method:[/underline][/italic] 728 women with no history of genital surgery (18-69 yrs) rated the sexual pleasure, discomfort, intensity of orgasm and effort required for achieving orgasm in specified areas around the clitoris and within the vagina, as well as appearance, size and position of clitoris and vagina (Dutch translation of the Self-Assessment of Genital Anatomy and Sexual Function- Female (1)). Anatomical areas were compared for the functional ratings by repeated measures ANOVA. [italic][underline]Results:[/underline][/italic] Anatomically, 45.2% of the sample described their clitoris as [apos]moderate-sized and raised[apos], and 43.4 % as [apos]small and raised[apos]. 88.2% reported their vagina to be [apos]adequate for sexual penetration[apos]. Sexual pleasure and orgasm was strongest and effort to attain orgasm and discomfort lowest when stimulating the clitoris and sides of the clitoris. Ratings of overall clitoral sensitivity to sexual stimulation gave higher values of sexual pleasure and discomfort for stimulation by the partner(p[lt]0.001). Orgasm intensity was higher and effort to achieve orgasm lower for clitoral stimulation by self compared to partner stimulation (p[lt]0.001). For vaginal sensitivity, scores for sexual pleasure, discomfort, orgasm intensity and effort increased with increasing vaginal depth. Decreased orgasmic sensitivity was indicated by lower scores for the vagina (on all depth levels) than for the clitoris (p[lt] 0.001). [italic][underline]Conclusion:[underline][/italic] The SAGAS-F discriminates well between various genital areas in terms of erotic sensitivity, when administered to genitally unoperated women varying in age, parity status and socio-economic level. The clitoris itself appeared to be most sensitive, which seems to be consistent with maximum nerve density in this area. Clitoral surgery could disrupt neurological pathways and compromise erotic sensation and pleasure. Data on sexual function and sensation should form part of the long-term follow-up of genitoplasty procedures.[br][br](1)Schober,J.M. et al, BJU Int 2004; 94:589-594.[br][br]Sources of Research Support: FWO Vlaanderen.[br][br]Nothing to Disclose: GB, NC, JMS, HFLM-B, GDC, GT, MC, PH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1275 294 2010 MON-60 PO37-01 Monday 1678 2012


1675 ENDO12L_MON-61 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) Monogenic Disorders in 46, XY Disorders of Sex Development with Genital Anomalies: Comprehensive Genetic Study in Japan Mie Hayashi, Tomohiro Ishii, Fumiko Kato, Ayuko Suwanai, Masaki Takagi, Keiko Homma, Hiroshi Asanuma, Maki Fukami, Hirofumi Oohashi, Tomonobu Hasegawa Keio University School of Medicine, Tokyo, Japan; Keio University School of Medicine, Tokyo, Japan; Keio University School of Medicine, Tokyo, Japan; National Research Institute for Child Health and Development, Tokyo, Japan; Saitama Children[apos]s Medical Center, Saitama, Japan [bold]Background[/bold]: Eleven genes have been proven to be responsible for monogenic disorders in 46,XY disorders of sex development (DSD) with genital anomalies. As far as we know, no comprehensive genetic study has been reported including possible candidate genes.[br][bold]Objective[/bold]: The objectives of this study was to evaluate the frequency of monogenic disorders in Japanese patients having 46,XY DSD with genital anomalies.[br][bold]Subjects and Methods[/bold]: We analyzed 93 Japanese index patients with 46,XY DSD with genital anomalies for the following eighteen responsible or candidate genes by PCR direct-sequence; [italic]SRY, NR5A1, DAX1, DHH, LHX9, WT1, MAMLD1, CBX2, GATA4, DMRT1, SOX9, [/italic][italic]AR[/italic], [italic]SRD5A2[/italic], [italic]STAR[/italic], [italic]CYP11A1[/italic], [italic]CYP17A1[/italic], [italic]HSD3B2[/italic] and [italic]HSD17B3[/italic]. We studied pathogenicity of putative novel mutations [italic]in vitro [/italic]and [italic]in silico[/italic].[br][bold]Results[/bold]: We identified ten non synonymous sequence variations; [italic]S[/italic][italic]F1[/italic] p.C55F, p.D257fsX39, [italic]SF1[/italic] p.V424del, [italic]SRY[/italic] p.R75S, [italic]MAMLD1[/italic] p.Q419_Q421del[italic], AR[/italic] p.A597T, AR p.D604N, AR p.Q825K, [italic]SRD5A2[/italic] p.N193S/R227E, [italic]SRD5A2[/italic] p.R103X/R246Q. Seven of ten mutations were recurrent. None of the other 3 variations (NR5A1 p.C55F, MAMLD1 p.Q419_Q421del and AR p.D605N) were detected in normal 150 male controls. Functional analyses demonstrated all these three were loss-of-function mutations. The frequency of the monogenic mutations was 11% [95% Confidential Interval (CI) 5.3-18.9%]; [italic]SF1, AR[/italic] 3.3% [0.67-9.1%], [italic]SRD5A2[/italic]; 2.2% [0.26-7.6%], [italic]SRY[/italic] and [italic]MAMLD1[/italic], 1.1% [0.27-9.1%].[br][bold]Conclusions[/bold]: We first described the frequency of monogenic disorders was 11%(95% confidence interval, 5.3 to 18.8) in 46, XY DSD with genital anomalies in [italic]SRY, NR5A1, DAX1, DHH, LHX9, WT1, MAMLD1, CBX2, GATA4, DMRT1, SOX9, [/italic][italic]AR[/italic], [italic]SRD5A2[/italic], [italic]STAR[/italic], [italic]CYP11A1[/italic], [italic]CYP17A1[/italic], [italic]HSD3B2[/italic] and [italic]HSD17B3 [/italic].[br][br]Nothing to Disclose: MH, TI, FK, AS, MT, KH, HA, MF, HO, TH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1159 294 2011 MON-61 PO37-01 Monday 1679 2012


1676 ENDO12L_MON-62 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) Ten Novel Mutations in the [italic]NR5A1[/italic] Gene Cause Disordered Sex Development in 46,XY and Ovarian Insufficiency in 46,XX Individuals Nuria Camats, Amit V Pandey, Monica Fernandez-Cancio, Pilar Andaluz, Marco Janner, Nuria Toran, Primus E Mullis, Antonio Carrascosa, Laura Audi, Christa E Fluck University Children[apos]s Hospital Bern, Bern, Switzerland; Hospital Universitari Vall d[apos]Hebron, Barcelona, Spain; Hospital Universitari Vall d[apos]Hebron, Barcelona, Spain [bold]Introduction.[/bold][br]Steroidogenic factor-1 (SF-1), encoded by the [italic]NR5A1[/italic] gene, is a nuclear receptor which regulates many aspects of adrenal and reproductive development and function. [italic]NR5A1[/italic] mutations have been detected in 46,XY individuals with disorders of sex development (DSD) but apparently normal adrenal function and in 46,XX women with normal sexual development yet primary ovarian insufficiency (POI). Our aim was to study a group of 100 46,XY DSD and 2 POI patients for [italic]NR5A1[/italic] mutations and its impact on phenotype, tissue histology and [italic]in vivo[/italic] versus [italic]in vitro[/italic] function.[br][bold]Methods.[/bold][br]Clinical, biochemical, histological, genetic and functional analyses were performed. Patients were referred from different centres in Spain (65 46,XY DSD), Switzerland (2 POI) and Turkey (35 46,XY DSD). Histologic studies were performed. Genetic studies involved the sequencing and analysis of the 5[apos]UTR region and the 7 exons with their flanking intronic sequences. Functional analyses were performed in steroidogenic adrenal NCI-H295R and non-steroidogenic HEK293 cells. Cells were transiently transfected with wild-type and mutant SF-1 and known SF-1 regulated promoter luciferase reporter constructs. Finally, phenotype-genotype and structure-function correlations were assessed.[br][bold]Results.[/bold][br]Ten novel heterozygote [italic]NR5A1[/italic] mutations were detected (5 missense, 1 nonsense, 3 frameshift mutations and 1 duplication); these were scattered throughout exons 2 to 5. Clinical phenotype varies not only widely with specific mutations but also with patients harboring an identical SF-1 mutation. Histologic studies of testes showed vacuolization of Leydig cells due to fat accumulation with age. Promoter transactivation assays revealed that SF-1 mutations located in the DNA binding domain of the protein grossly reduced transactivation activity while mutations in other domains showed mixed results. Results were also variable according to cell systems and specific promoters analysed. In line with high variability in functional assays, we were also not able to find structure-function correlation when using bioinformatic tools.[br][bold]Conclusions.[/bold][br][italic]NR5A1[/italic] mutations are frequently found in 46,XY DSD individuals (9%) and manifest with a broad phenotype. Testes histology is characteristic for fat accumulation and degeneration over time similar to findings observed in patients with lipoid congenital adrenal hyperplasia (due to [italic]StAR[/italic] mutations). Genotype-structure-function-phenotype correlation remains still elusive.[br][br]Nothing to Disclose: NC, AVP, MF-C, PA, MJ, NT, PEM, AC, LA, CEF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 496 294 2012 MON-62 PO37-01 Monday 1680 2012


1677 ENDO12L_MON-63 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) A Novel Homozygous Missense Mutation in Steroidogenic Factor 1 Causes Complete 46,XY Sex Reversal and Asplenia Ehud Banne, Yotam Kaufman, Ariella Weinberg-Shukron, Abdulsalam Abulibdeh, Yackov Berkun, Ephrat Levy-Lahad, David Zangen Hadassah Hebrew University Medical Center, Jerusalem, Israel; Hebrew University Medical School, Jerusalem, Israel; Shaare Zedek Medical Center, Jerusalem, Israel; Hadassah Mount Scopus, Hadassah Hebrew University Medical Center, Jerusalem, Israel [bold]Background[/bold]: Steroidogenic Factor 1 (SF-1) is important for steroidogenic and nonsteroidogenic tissue differentiation. Most reported SF-1 mutaions are heterozygous and result in mild to severe Disorders of Sex Development (DSD). We describe a unique clinical presentation of a novel homozygous SF-1 mutation.[br][bold]Patient and Methods:[/bold] Following routine ultrasonographic studies for abdominal pain revealling asplenia and the absence of a uterus in a 13.5 prepubertal usually healthy girl; karyotype was found to be 46XY. Further imaging, adrenocortical function tests, testicular histology studies and molecular sequencing of the StAR and SF-1 genes were performed.[br][bold]Results[/bold]: CT studies confirmed asplenia, absence of uterus and ovaries, with gonads (testis) present in the inguinal regions. Peripheral blood smear revealed Howell Jolly bodies and pokilocytosis characterizing asplenia. Electrolytes, aldosterone as well as basal and ACTH stimulated cortisol, and 17[ndash]OH-progesterone were normal. Testicular histopathology revealed absence of germinal cells, no preneoplastic markers and very few Leidig cells. A homozygous G to A mutation in a cross species conserved region in exon 4 of the SF1 gene was identified, resulting in a non tolerable (SIFT program) R103Q amino acid substitution.[br][bold]Conclusions: [/bold]This novel homozygous SF-1 mutation is the first XY-DSD case worldwide associated with human asplenia, mimicking the asplenia in SF-1 knockout mice. The specific absence of germinal precursors and Leidig cells indicates that the crucial SF-1 mutation prevents their differentiation but enables the presence and secretory function of Sertoli cells. The variety of phenotypes in SF-1 heterozygous mutations singles out this [ldquo]clean[rdquo] homozygous presentation illustrating the important role of the F1 box domain of SF-1 in severe DSD and asplenia. Asplenic and alternatively DSD patients should be screened for the reciprocal childhood asymptomatic condition.[br][br]Nothing to Disclose: EB, YK, AW-S, AA, YB, EL-L, DZ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1186 294 2013 MON-63 PO37-01 Monday 1681 2012


1678 ENDO12L_MON-64 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) Clinical, Molecular and Functional Characteristics of a Novel Mutation in the [italic]CYP17A1[/italic] Gene in Patients with Combined 17[alpha]-Hydroxylase/17,20-Lyase Deficiency Yoo-Mi Kim, Chang-Woo Jung, Minji Kang, Gu-Hwan Kim, Beom Hee Lee, Jin-Ho Choi, Jung Hun Ohn, Ju Hee Lee, Jung Hee Kim, Seong Yeon Kim, Han-Wook Yoo Asan Medical Center, Seoul, Republic of Korea; Asan Medical Center, Seoul, Republic of Korea; Asan Medical Center, Seoul, Republic of Korea; Seoul National University College of Medicine, Seoul, Republic of Korea [bold]Purpose:[/bold] Combined 17[alpha]-hydroxylase/17,20-lyase deficiency is a rare autosomal recessive form of congenital adrenal hyperplasia presenting with hypertension and sexual infantilism. This disorder is caused by defects in P450c17, encoded by the [italic]CYP17A1[/italic] gene. The goal of this study is to describe the clinical and molecular characteristics of five patients with 17[alpha]-hydroxylase/17,20-lyase deficiency, and investigate functional alterations of the CYP17A1 mutants.[br][bold]Subjects and methods:[/bold] Five patients were evaluated for primary amenorrhea and hypertension. The median age at diagnosis was 20.0 years (range, 11-48 years). Karyotypes were 46, XY in three patients and 46, XX in the other patients. In all patients, serum gonadotropin, ACTH, progesterone, and 11-deoxycorticosterone levels were elevated. However, testosterone and dehydroepiandrosterone sulfate (DHEA-S) levels were low. The coding regions of the [italic]CYP17A1[/italic] gene were amplified by PCR and directly sequenced. Mutant cDNA was constructed by site-directed mutagenesis. Wild-type and mutant [italic]CYP17A1[/italic] cDNA was inserted into expression vector, pcDNA3.1-V5/His-P450c17, and transiently expressed in COS-7 cells. The 17[alpha]-hydroxylase and 17,20-lyase activities were assayed by examining the conversions of progesterone to 17-hydroxyprogesterone and 17-hydroxypregnenolone to DHEA using radioimmunoassay.[br][bold]Results:[/bold] In subject 1, sequencing of the [italic]CYP17A1[/italic] identified a compound heterozygosity consisting of p.H373L and p.W406L. Subject 2 was found to be homozygous for p.H373L mutation. The other three patients were heterozygous for p.Y329fs. To assess the functional consequences of the novel p.W406L mutation, COS-7 cells were transfected with the expression vector pcDNA3.1-V5/His-P450c17 containing either wild-type or mutant [italic]CYP17A1[/italic] cDNA. The p.W406L mutant protein expressed in COS-7 cells showed a complete loss of 17[alpha]-hydroxylase as well as 17,20-lyase activities compared to wild-type protein.[br][bold]Conclusions: [/bold]This is the first study describing the clinical and molecular characteristics of 17[alpha]-hydroxylase/17,20-lyase deficiency in Korea. The novel P450c17 mutation p.W406L abolished both enzyme activities. The complete loss of both 17[alpha]-hydroxylase and 17,20-lyase activities in p.W406L indicates that this locus is essential for enzyme activity.[br][br]Nothing to Disclose: Y-MK, C-WJ, MK, G-HK, BL, J-HC, JHO, JHL, JHK, SYK, H-WY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1145 294 2014 MON-64 PO37-01 Monday 1682 2012


1679 ENDO12L_MON-65 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) A Large Insertion in the Promoter Region of the Androgen Receptor Gene: A Rare Genetic Cause of Partial Androgen Insensitivity Syndrome Andresa DS Rodrigues, Natalia P Pareira, Mirian Nishi, Karla F Melo, Mariana Funari, Sorahia Domenice, Ivo JP Arnhold, Berenice Bilharinho Mendonca, Elaine MF Costa Faculdade de Medicina da Universidade de Sao Paulo, Sao Paulo, Brazil Over 400 mutations in the Androgen Receptor gene (AR) causing Androgen Insensitivity syndrome (AIS) have been described. The frequency of deletions or insertions in the AR coding region is around 5-10%. In our cohort of Brazilian patients with AIS, missense or nonsense mutations were identified in all patients with the complete form (CAIS) and in 70% of those with partial form (PAIS), suggesting that the genetic alterations, responsible for the phenotype, might be located in AR non-coding regions or in other genes related to androgen receptor activity. Objective: to analyze the AR non-coding region in a family with PAIS and normal sequence in the coding region. Patients: The family is composed of 9 PAIS affected members with typical X-linked inheritance. The AR entire coding region had been previously studied in 5 of the 9 patients and no mutation was identified. However, the identification of 21 CAG repeats in exon 1 in all affected members strongly suggested that the AR was involved in the etiology of PAIS in this family. Methods: cDNA obtained from mRNA extracted from skin fibroblasts culture was directly sequenced in one affected male (propositus) and qPCR was carried out using the TaqMan assay with Hs00171172 and CYC, GAPDH, PGK-1 and B2M as endogenous genes. mRNA from skin fibroblasts from a normal individual was used as control. Direct sequencing of the AR promoter region in genomic DNA of the propositus was performed. Results: No allelic variants in AR cDNA were found. qPCR analysis identified a very low expression (10%) of the AR compared with the control. Direct sequencing of the promoter region indicated an approx. 1000 pb insertion at [ndash] 285 pb position. However, the conventional direct sequencing technique was not able to read the base sequence of the fragment because it was rich in A/T. Discussion: We describe a large Brazilian family with 9 PAIS affected members without mutation in the coding region and a strong evidence of implication of the AR in the etiology of the disease. Normal cDNA of the propositus ruled out splice site alterations. On the other hand, the detection of low expression of the AR led us to analyze the promoter region of the gene. Conclusion: This is the first report of a large insertion in the AR promoter region causing PAIS identified in a Brazilian family with 9 affected members. The complete characterization of the new AR promoter region alteration will expand the repertoire of mutations in the AR gene.[br][br]Nothing to Disclose: ADSR, NPP, MN, KFM, MF, SD, IJPA, BBM, EMFC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 756 294 2015 MON-65 PO37-01 Monday 1683 2012


1680 ENDO12L_MON-66 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) Screening for Microdeletions in the AZF Region of the Y Chromosome in Patients with Disorders of Sex Development Due to 45,X/46,XY Chromosome Abnormalities Mirian Yumie Nishi, Priscila Tamires Lima Santos, Elaine Maria Frade Costa, Berenice Bilharinho Mendonca, Sorahia Domenice Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil The etiology of the disorders of sex development (DSD) in patients with 45,X/46,XY karyotype and variants is not yet completely understood. The presence of Yq microdeletions in men with azoospermia or severe oligospermia are frequent, but recently these deletions have also been identified in subjects with Klinefelter syndrome (KS) and with 45,X/46,XY mosaicisms. It has been hypothesized that deletions of the Azoospermia factor region (AZFa, AZFb and AZFc sub-regions), located at Yq, might predispose to Y loss. Deletions of AZFc, which includes the [italic]DAZ[/italic] genes, are a common cause of spermatogenic failure. [bold]Objective[/bold]: to screen Yq microdeletions in Brazilian patients with DSD due to 45,X/46,XY chromosomal abnormalities. [bold]Patients and[/bold] [bold]Methods[/bold]: Twenty-seven subjects with 45,X/46,XY or with 45,X/46,X,idic(Y) karyotypes were selected, 16 with mixed gonadal dysgenesis (MGD) and 11 with Turner syndrome (TS). DYZ3 (centromere) and seven [italic]loci[/italic] covering AZF regions were screened using PCR in DNA from blood. The [italic]loci[/italic] DYS280 and [italic]UTY[/italic] are located in AZFa; DYS216, DYS231 and DYS224 in AZFb; [italic]DAZ[/italic] and [italic]PPP1R12BP1[/italic] in AZFc regions. [bold]Results[/bold]: Yq microdeletions were detected in 6 (22%) patients: 3 with MGD and 3 with TS. Regarding MGD patients, in one it spans at least 6 Mb (DYS216 to [italic]PPP1R12BP1[/italic]), in the second 4.5 Mb (DYS231 to [italic]PPP1R12BP1[/italic]), and in the third one as so as in the 3 patients with TS they span at least 3 Mb ([italic]DAZ[/italic] to [italic]PPP1R12BP1[/italic]). [bold]Discussion[/bold]: The Yq deletions identified in DNA of these patients involve the AZFb and AZFc regions. The longest deletions of Yq, containing AZFb and AZFc regions, were identified in two MGD patients with male phenotype. The AZFc region was deleted in all 6 patients. Likely, the deletions of AZFc region are the most common identified in patients with idiopathic infertility due to oligozoospermia or complete absence of germ cells as in KS. In 45,X/46,XY patients, there are a few reports identifying deletions in AZFb beyond in the AZFc region as in the present study (1,2). Alvarez-Nava et [italic]al[/italic].(3) studying gonadal tissues from 45,X/46,XY patients, identified a higher incidence of Yq microdeletions in dysgenetic gonads than in blood. [bold]Conclusion[/bold]: Extensive studies are needed to establish the exact association between specific Yq microdeletions and the various degrees of gonadal dysgenesis in patients with DSD due to 45,X/46,XY chromosome abnormalities and to confirm the role of this mechanism for the formation of 45,X cell lines in TS and in MGD patients.[br][br](1)Siffroid JP et al., Human Reprod 2000; 15:2559. (2)Patsalis PC et al., Am J Med Genet 2005; 135:145. (2)Alvarez-Nava F et al., Fert Steril 2008; 89:458.[br][br]Sources of Research Support: CNPq 305743/2011-2 awarded to MBB.[br][br]Nothing to Disclose: MYN, PTLS, EMFC, BBM, SD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1452 294 2016 MON-66 PO37-01 Monday 1684 2012


1681 ENDO12L_MON-67 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) Excess DAX1 Leads to XY Ovotesticular DSD in Mice by Inhibiting SF1 Activation of the Testis Enhancer of [italic]Sox9[/italic] Louisa M Ludbrook, Pascal Bernard, Stefan Bagheri-Fam, Ryohei Sekido, Dagmar Wilhelm, Robin Lovell-Badge, Vincent Russel Harley Prince Henry[apos]s Institute of Medical Research, Clayton Melbourne, Australia; Monash University, Clayton Melbourne, Australia; Monash University, Clayton Melbourne, Australia; MRC National Institute for Medical Research, Mill Hill, London, UK; University of Queensland, St Lucia, Australia Human [italic]DAX1[/italic] duplications cause Dosage-Sensitive Sex reversal (DSS) whereby chromosomally XY individuals can develop as females, due to gonadal dysgenesis. We show that embryonic testes from XY [italic]Dax1[/italic]-overexpressing transgenic mice are underdeveloped and expression of the key testis-promoting gene [italic]Sox9[/italic] is reduced. In XY [italic]Sox9[/italic] heterozygotes, where testis development is usually normal, [italic]Dax1[/italic] overexpression results in ovotestes, suggesting a DAX1-SOX9 antagonism. Using reporter mice, [italic]Dax1[/italic] overexpression reduced activation of [italic]TES[/italic], the testis enhancer of [italic]Sox9[/italic], known to integrate the activity of several transcription factors. Increasing levels of DAX1 specifically antagonised SF1-mediated activation of both mouse and human [italic]TES in vitro[/italic], providing a likely mechanism for DSS.[br][br]Sources of Research Support: NHMRC Program Grants 334314 and 546517 (VH), Australian Postgraduate Award (LL) and Monash University Travel Award (LL), and the UK Medical Research Council (U117512772) (RLB).[br][br]Nothing to Disclose: LML, PB, SB-F, RS, DW, RL-B, VRH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 27 294 2017 MON-67 PO37-01 Monday 1685 2012


1682 ENDO12L_MON-68 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) Delay in the Onset of Puberty of Intrauterine Growth Retarded Female Rats Cannot Be Rescued with Hypernutrition after Birth Toshiya Matsuzaki, Ganbat Gereltsetseg, Takeshi Iwasa, Riyo Kinouchi, Hiroshi Nakazawa, Minoru Irahara The University of Tokushima Graduate School, Tokushima, Japan Perinatal undernutrition is known to disturb reproductive development, in particular by delaying the onset of puberty in certain species. Using a rat model, we studied whether hypernutrition after birth can rescue the delayed onset of puberty in intrauterine undernourished female rats. Pregnant rats were divided into two groups: the maternal normal nutrition (mNN, n = 8) and maternal undernutrition (mUN, n = 9) groups. In the mUN group, dams received 50% of the daily food intake of the mNN group from day 15 of pregnancy until delivery. Pups from both the mNN and mUN dams were then separated into two groups, based on their postnatal feeding conditions: control-normal nutrition (control-NN), control-hypernutrition (control-HN), IUGR-normal nutrition (IUGR-NN), and IUGR-hypernutrition (IUGR-HN). Litter sizes of the hypernutrition groups were controlled to five pups per dam, and normal nutrition groups to 12-13 pups per dam. From postnatal day 30, pups were inspected daily for vaginal opening (VO). The age of VO in the IUGR-NN group was 35.7 [plusmn] 2.4 days (mean [plusmn] SD), which was significantly delayed compared to that of the control-NN group (33.8 0.8 days). The age of VO in the IUGR-HN group was 35.5 [plusmn] 2.3 days, which was significantly delayed compared to that of the control-HN group (33.5 [plusmn] 0.8 days). Interestingly, the age of VO did not differ between the IUGR-NN and IUGR-HN groups. In conclusion, maternal undernutrition delays puberty in female offspring, and this delay in puberty cannot be rescued with hypernutrition after birth.[br][br]Nothing to Disclose: TM, GG, TI, RK, HN, MI 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 298 294 2018 MON-68 PO37-01 Monday 1686 2012


1683 ENDO12L_MON-69 POSTER SESSION: Sex Determination [amp] Differentiation (1:30 PM-3:30 PM) Early Postnatal Lipopolysaccharide Exposure: Effects on Adult Male Rat LH Response to Homotypic Stress Hiroshi Nakazawa, Toshiya Matsuzaki, Takeshi Iwasa, Riyo Kinouchi, Ganbat Gereltsetseg, Minoru Irahara The University of Tokushima Graduate School, Tokushima, Japan Early-life immune stress possibly has long-lasting effects, called programming effects, on physiological responses to stress in adulthood; there may be a critical window during which such a challenge can induce long-lasting alterations. There have been few reports regarding this phenomenon in reproductive function. Here we report on induction by early-life LPS injection of long-lasting alterations in adult LH response to homotypic immune stress in male rats. Prior to this study, we investigated developmental changes in LH response to LPS, since immune challenge during the stress hyporesponsive period can induce long-lasting effects on physiological functions. Rat serum LH concentrations were decreased by LPS (100 [mu]g/kg) injection on postnatal days 15 and 25, but not 10, suggesting that the period prior to postnatal day 10 is the stress hyporesponsive period for LH. Serum LH concentrations and body weight were decreased by adult LPS (400 [mu]g/kg) injection in rats given saline or LPS (100 [mu]g/kg) on postnatal day 25, but not in rats given LPS (100[mu]g/kg) on postnatal day 10. Expression of hypothalamic IL-1[beta] and TNF-[alpha] mRNA, which suppress serum LH during immune stress, were equally increased in these groups by adult LPS (400 [mu]g/kg) injection. The present data suggest that the period prior to postnatal day 10 is the critical window in which immune stress can induce long-lasting alterations in the LH response, and that IL-1[beta] and TNF-[alpha] might not be involved, in this experimental model, in this alteration in rats given LPS on postnatal day 10.[br][br]Nothing to Disclose: HN, TM, TI, RK, GG, MI 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 297 294 2019 MON-69 PO37-01 Monday 1687 2012


1684 ENDO12L_MON-85 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Meal Timing and Composition Influence Ghrelin Levels, Appetite Scores and Weight Loss Maintenance in Overweight and Obese Adults Daniela Jakubowicz, Oren Froy, Julio Wainstein, Mona Boaz Wolfson Medical Center, Tel Aviv University, Holon, Israel; The Hebrew University of Jerusalem, Rehovot, Israel; Wolfson Medical Center, Tel Aviv University, Holon, Israel; Wolfson Medical Center, Tel Aviv University, Holon, Israel [bold]Background: [/bold]Although dietary restriction often results in initial weight loss, the majority of obese dieters fail to maintain their reduced weight. Diet-induced weight loss results in compensatory increase of hunger, craving and decreased ghrelin suppression that encourage weight regain. A high protein and carbohydrate breakfast may overcome these compensatory changes and prevent obesity relapse.[br][bold]Methods: [/bold]In this study 193 obese (BMI 32.2[plusmn]1.0 kg/m[sup]2[/sup]), sedentary non diabetic adult men and women (47[plusmn]7years) were randomized to a low carbohydrate breakfast (LCb) or an isocaloric diet with high carbohydrate and protein breakfast (HCPb). Anthropometric measures were assessed every 4 weeks. Fasting glucose, insulin, ghrelin, lipids, craving scores and breakfast meal challenge assessing hunger, satiety, insulin and ghrelin responses, were performed at baseline, after a diet Intervention period (week 16) and after a follow-up period (week 32).[br][bold]Results: [/bold]At week16, groups exhibited similar weight loss:-15.1[plusmn]1.9 kg in LCb group vs.13.5[plusmn]2.3 kg in HCPb group, p=0.11. From week 16 to week 32, LCb group regained 11.6 [plusmn]2.6 kg, while the HCPb group lost additional 6.9[plusmn]1.7 kg. Ghrelin levels were reduced after breakfast by 45.2% and 29.5% following the HCPb and LCb, respectively. Satiety was significantly improved and hunger and craving scores significantly reduced in the HCPb group vs. the LCb group.[br][bold]Conclusion: [/bold][br]A high carbohydrate and protein breakfast may prevent weight regain by reducing diet-induced compensatory changes in hunger, cravings and ghrelin suppression.[br]To achieve long-term weight loss, meal timing and macronutrient composition must counteract these compensatory mechanisms which encourage weight regain after weight loss.[br][br]Nothing to Disclose: DJ, OF, JW, MB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 17 295 2020 MON-85 PO27-03 Monday 1688 2012


1685 ENDO12L_MON-86 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) The Effects of Sham Feeding and GLP-1 Receptor Blockade on Subsequent Food Intake in Patients with Gastric Bypass Surgery Jason G Barrera, Leslie M Baum, Stephen C Woods, David A D[apos]Alessio, Marzieh Salehi University of Cincinnati, Cincinnati, OH; University of Cincinnati, Cincinnati, OH Roux-en-Y gastric bypass (GB) surgery is associated with significant and durable weight loss. While the mechanisms by which GB affects energy balance are not entirely clear, studies of non-surgical (NS) subjects suggest that preprandial neural and hormonal anticipatory signals may influence energy homeostasis and food intake. These effects may be amplified in GB subjects.[br]To test this hypothesis, we measured ad libitum caloric intake of a mixed meal in groups of GB (N=5) and nonsurgical (NS, n=8) subjects 3 hours after ingestion of a liquid test meal (Ensure plus, 300 kcal) on three separate days: a) control (saline), b) sham feeding (chewing and expectorating a peanut butter sandwich, 255 kcal) preceding the test meal, and c) following 4-hour iv infusion of the glucagon-like peptide-1 receptor (GLP-1r) antagonist exendin (9-39) (Ex9, 750 pg/kg*min). Surgical patients were older and thinner (age, 50 vs. 34 yrs; BMI, 30 vs. 35 kg/m[sup]2[/sup]) and were on average 3.4 years out from surgery (range 2-5 years). Subjects had access to food for 45 min and were blinded to the measurement of food intake during the post-study lunch. Caloric intake was compared to control for the sham feeding and Ex9 studies separately, using two-way repeated measures ANOVA.[br]Caloric intake was significantly lower in GB vs. NS subjects (p[lt]0.05); this difference was similar following saline (GB 630; NS 1116 kcal), sham feeding (GB 481; NS 823 kcal), and Ex9 infusion (GB 552; NS 926 kcal) (p[lt]0.05 for all). Sham feeding was associated with a lower food intake at lunch (p=0.029) in both GB (saline 696; sham 481 kcal) and NS (saline 1074; sham 823 kcal) subjects. There was a trend toward reduced total caloric intake in Ex9- vs. saline-treated subjects (p=0.072) with no significant difference between NS (saline 1116; Ex9 926 kcal) vs. GB (saline 630; Ex9 552 kcal) subjects.[br]These data indicate that caloric intake at subsequent meals is partly accounted for by neural responses to sham feeding as well as preprandial GLP-1 action. Although subjects with GB had a lower baseline caloric intake, their responses to sham feeding and GLP-1 receptor blockade were not different from those of control subjects, indicating that these factors are not magnified in GB subjects and thus may not account for GB-associated weight loss.[br][br]Sources of Research Support: Grants from National Institutes of Health, DK083554 (MS) and in part by USPHS, UL1 RR026314 from the National Center for Research Resources, NIH.[br][br]Nothing to Disclose: JGB, LMB, SCW, DAD, MS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1613 295 2021 MON-86 PO27-03 Monday 1689 2012


1686 ENDO12L_MON-87 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Rapid and Strong Suppression of Human Acylated Ghrelin Serum Concentrations during Infusion of Des-Acyl Ghrelin in Obese Diabetic Subjects Behiye Ozcan, Sebastian JCMM Neggers, Anne Reifel Miller, H-C Yang, Patrick Delhanty, Soraya Allas, Thierry Abribat, A J van der Lely Erasmus University Medical Centre Rotterdam, Rotterdam, Netherlands; Aliz[eacute] Pharma SA, Ecully, France; Lilly Corporate Center, Indianapolis, IN Ghrelin, a gut hormone, which can be found in two forms in the body. As acylated form (AG) and unacylated form (UAG). UAG sometimes agonizing, sometimes antagonizing the effects of AG. In particular, UAG is able to prevent the hyperglycemic effects of ghrelin, when administered concomitantly, in healthy volunteers and there are sereval reports on the anti-diabetogenic potentials of UAG (1-5).[br]Aim[br]To assess the effects of a continuous overnight infusion of two doses of UAG (3[amp]10 [mu]g UAG/kg.hr-1) versus placebo in a double blind cross-over study on AG levels, as well on glucose and insulin response to a standard breakfast meal (SBM) in 8 overweight patients with type 2 diabetes with acceptable metabolic control (HbA1c 6.5-7.5%).[br]Methods[br]During these 3 admissions, in randomized order, all subjects received either placebo, or UAG 3 [mu]g/kg.hr-1 or UAG 10 [mu]g/kg.hr-1 as a continuous venous infusion. The admission were divided by a wash-out period of [gt]7 days. Serum glucose were assessed at regular intervals and via a continuous glucose monitor. Continuous infusion of either placebo or UAG started at 22.00 until 13.00 hrs the next day. In the morning between 8.00-8.15 hrs. a standard breakfast meal (SBM) was used. SBM consisted out of 714 kcal (17% proteins; 46 % fat; 37 % carbohydrates).[br]Results[br]Morning AG levels (before SBM) dropped significantly from 21.01[plusmn]8.9 (mean[plusmn]SD) to 3.0[plusmn]6.7, to 1.4[plusmn]3.2 pg/ml after placebo, 3 and 10 mcg/kg.hr-1 UAG infusion, respectively. After breakfast AG levels drop from 14.03[plusmn]9.4, 0.8[plusmn]1.8, to 0.8[plusmn]1.8 pg/ml after placebo, 3 and 10 mcg/kg.hr-1 UAG infusion, respectively.[br]UAG levels, increased from 105.9[plusmn]31.4, 10,998[plusmn]2,623, 12,085[plusmn]1,616 pg/ml after placebo, 3 and 10 mcg/kg.hr-1 UAG infusion, respectively.[br]During continuous glucose monitoring (CGMS), post-SBM, overnight infusion of 10 mcg/kg.hr UAG significantly decreased glucose levels. The AUC decreased from 1618, 1601, to 1540mmol/3 hrs of placebo, 3 and 10 mcg UAG infusion.[br]A significant decrease in peak serum glucose levels after SBM during 10 mcg UAG infusion was observed.[br]So, UAG is a potent inhibitor of ghrelin levels in obese human subjects with mild diabetes. Additionaly, UAG improves postprandial glycemia, especially in those subjects in whom preprandial acylated ghrelin levels are high, which makes UAG (analogs) strong potential candidates for the development of drugs in the treatment of metabolic disorders or other conditions in which a disturbed AG/UAG ratio has been found.[br][br]1 Kiewiet RM, van Aken MO, van der Weerd K, Uitterlinden P, Themmen AP, Hofland LJ, de Rijke YB, Delhanty PJ, Ghigo E, Abribat T, van der Lely AJ 2009 Effects of acute administration of acylated and unacylated ghrelin on glucose and insulin concentrations in morbidly obese subjects without overt diabetes. Eur J Endocrinol 161:567-573. 2 Gauna C, Kiewiet RM, Janssen JA, van de ZB, Delhanty PJ, Ghigo E, Hofland LJ, Themmen AP, van der Lely AJ 2007 Unacylated ghrelin acts as a potent insulin secretagogue in glucose-stimulated conditions. AmJPhysiol EndocrinolMetab 293:E697-E704. 3 Broglio F, Gottero C, Prodam F, Gauna C, Muccioli G, Papotti M, Abribat T, van der Lely AJ, Ghigo E 2004 Non-acylated ghrelin counteracts the metabolic but not the neuroendocrine response to acylated ghrelin in humans. Journal of Clinical Endocrinology [amp] Metabolism 89:3062-3065. 4 Gauna C, Meyler FM, Janssen J, Delhanty PJD, Abribat T, Van Koetsveld P, Hofland LJ, Broglio F, Ghigo E, van der Lely AJ 2004 Administration of acylated ghrelin reduces insulin sensitivity, whereas the combination of acylated plus unacylated ghrelin strongly improves insulin sensitivity. Journal of Clinical Endocrinology and Metabolism 89:5035-5042. 5 Gauna C, Delhanty PJD, Hofland LJ, Janssen J, Broglio F, Ross RJM, Ghigo E, van der Lely AJ 2005 Ghrelin stimulates, whereas des-octanoyl ghrelin inhibits, glucose output by primary hepatocytes. Journal of Clinical Endocrinology and Metabolism 90:1055-1060.[br][br]Disclosures: ARM: Employee, Eli Lilly & Company. H-CY: Employee, Eli Lilly & Company. Nothing to Disclose: BO, SJCMMN, PD, SA, TA, AJvdL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 74 295 2022 MON-87 PO27-03 Monday 1690 2012


1687 ENDO12L_MON-88 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Importance of Brain Reward Circuitry in Adult Patients with Acquired Structural Hypothalamic Damage: A Functional Neuroimaging Study Joanne Powell, Caroline Steele, Ian A Macfarlane, Jason CG Halford, John PH Wilding, Jo A Harrold, Daniel J Cuthbertson, Kumar SV Das, Mohsen Javadpour, Graham J Kemp, Andrej Stancak, Christina Daousi University of Liverpool, Liverpool, UK; University of Liverpool, Liverpool, UK; The Walton Centre for Neurology and Neurosurgery, Liverpool, UK; University of Liverpool, Liverpool, UK [bold]Background: [/bold]Weight gain and obesity are common long-term consequences of hypothalamic damage due to hypothalamic tumors. The hypothalamus and brainstem are the two principal homeostatic brain areas responsible for regulating feeding behavior and body weight, whereas the corticolimbic, paralimbic and higher cortical brain regions process environmental cues and the reward properties of food. We hypothesised that reward-related brain regions may assume a more dominant role in appetite control in hypothalamic obesity (HO).[br][bold]Design and Subjects: [/bold]9 patients with HO [body mass index BMI[plusmn]SD 37.7[plusmn]5.4 kg/m[sup]2[/sup]; age 47[plusmn]15 y], 10 age-matched obese controls (OC) [BMI 38[plusmn]6 kg/m[sup]2[/sup]], 7 patients who remained weight-stable following hypothalamic insult (HWS) [BMI 26.9[plusmn]2.3 kg/m[sup]2[/sup], age 57[plusmn]17 y], and 10 non-obese controls (NOC) [BMI 26[plusmn]3kg/m[sup]2[/sup]] were studied. Functional magnetic resonance imaging (FMRI) was performed in the fasted state, one hour and 3 hours after consumption of a test meal, whilst subjects were presented with a series of visual stimuli. Each block of images portrayed either high calorie foods, low calorie foods or objects. FMR images were processed with Statistical Parametric Mapping software, followed by analysis of covariance to test for significant regional blood oxygen level-dependent (BOLD) response changes.[br][bold]Results: [/bold]Pictures of high calorie foods produced significantly greater BOLD activation in HO and OC subjects compared to NOC in the caudate head ([italic]P[/italic]=0.008), middle frontal gyrus ([italic]P[/italic]=0.021), insula ([italic]P[/italic]=0.032), cingulate gyrus ([italic]P[/italic]=0.03) and anterior cingulate gyrus ([italic]P[/italic][lt]0.001). Mean neuronal activation in the mid-insula in HWS patients was lower than in the other 3 groups (HWS=-0.042, HO=0.237, NOC=0.186, OC=0.175; [italic]P[/italic]=0.02). Activation of the lingual gyrus in HWS was also lower than all other 3 groups (HWS=1.309, HO=1.633, NOC=1.698 and OC=1.609; [italic]P[/italic]=0.025).[br][bold]Conclusions: [/bold]During viewing of high-calorie food-related stimuli, weight-stable patients with hypothalamic damage showed significantly less brain activation in regions linked to processing of interoceptive inputs such as the mid-insula and in regions linked to reward value for food such as the lingual gyrus. We have provided the first evidence that neural pathways that guide reward-related behavior in response to food may play a more important role in controlling eating behavior in patients with HO than in those who do not experience significant weight gain after hypothalamic damage.[br][br]Nothing to Disclose: JP, CS, IAM, JCGH, JPHW, JAH, DJC, KSVD, MJ, GJK, AS, CD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1246 295 2023 MON-88 PO27-03 Monday 1691 2012


1688 ENDO12L_MON-89 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Pro-Inflammatory wnt5a and Anti-Inflammatory sFRP5 Are Differentially Regulated by Nutritional Factors in Obese Human Subjects Dominik M Schulte, Nike Mueller, Katrin Neumann, Frank Oberhaeuser, Michael Faust, Heike Guedelhoefer, Wilhelm Krone, Matthias Laudes Universit[auml]tsklinikum Schleswig-Holstein, Campus Kiel, Kiel, Germany; Uniklinik K[ouml]ln, Cologne, Germany Background: Obesity is associated with macrophage infiltration of adipose tissue. These inflammatory cells affect adipocytes not only by classical cytokines but also by the secreted glycopeptide wnt5a. Healthy adipocytes on the other site are able to release the specific wnt5a inhibitor sFRP5. This protective effect, however, was found to be diminished in obesity. The aim of the present study was to examine (1) whether obese human subjects exhibit increased serum concentrations of wnt5a and (2) whether wnt5a and/or sFRP5 serum concentrations in obese subjects can be influenced by caloric restriction.[br]Methodology: 23 obese human subject (BMI 44.1[plusmn]1.1 kg/m2) and 12 age- and sex-matched lean controls (BMI 22.3[plusmn]0.4 kg/m2) were included into the study. Obese subjects were treated with a very-low-calorie-diet (approximately 800 kcal/d) for 12 weeks. Body composition was assessed by impedance analysis, insulin sensitivity was estimated by the leptin-to-adiponectin ratio and wnt5a and sFRP5 serum concentrations were measured by ELISA. sFRP5 expression in human adipose tissue biopsies was further determined on protein level by immunohistology.[br]Principal findings: Pro-inflammatory wnt5a was not measurable in any serum sample of lean control subjects. In patients with obesity, however, wnt5a became significantly detectable consistent with low-grade inflammation in such subjects. Caloric restriction resulted in a weight loss from 131.9[plusmn]4.0 to 112.3[plusmn]3.2 kg in the obese patients group. This was accompanied by a significant decrease of the leptin-to-adiponectin ratio, indicating improved insulin sensitivity. Interestingly, these metabolic improvements were associated with a significant increase in serum concentrations of the anti-inflammatory factor and wnt5a inhibitor sFRP5.[br]Conclusions/Significans: Obesity is associated with elevated serum levels of pro-inflammatory wnt5a in humans. Furthermore, caloric restriction beneficially affects serum concentrations of anti-inflammatory sFRP5 in such subjects. These findings suggest a novel regulatory system in low grade inflammation in obesity which can be influenced by nutritional therapy.[br][br]Nothing to Disclose: DMS, NM, KN, FO, MF, HG, WK, ML 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 13 295 2024 MON-89 PO27-03 Monday 1692 2012


1689 ENDO12L_MON-90 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Omentin-1 Secretion and Its Response to Acute and Chronic Energy Deprivation and to Leptin Administration in Men Ole-Petter Riksfjord Hamnvik, Bindiya Thakkar, Konstantinos Nikolaos Aronis, Christos Socrates Mantzoros Brigham and Women[apos]s Hospital and Harvard Medical School, Boston, MA; VA Boston Healthcare System and Harvard Medical School, Boston, MA; Boston Medical Center, Boston, MA; Beth Israel Deaconess Medical Center, Boston, MA Aim: Omentin-1 is an adipocytokine secreted predominantly by the stromal vascular cells of visceral adipose tissue (1). Omentin-1 has been shown to regulate several important metabolic functions, including insulin-stimulated glucose uptake in adipocytes (1). Levels of omentin-1 are inversely correlated with BMI and leptin (2) and one study has reported increased omentin-1 levels after medical weight loss (2). We investigated the circadian patterns of omentin-1 secretion, the effects of short-term fasting or long-term calorie deficit after bariatric surgery and the effect of leptin administration on omentin-1 levels.[br]Methods: 8 healthy men were admitted for 72 hours under three different conditions: Fed state, fasting state with placebo administration, or fasting state with metreleptin administration SC. Blood samples were collected every 15 minutes for the last 24 hours of each admission. In a separate study, blood was collected from 12 patients undergoing bariatric surgery before surgery and after 3 and 6 months. Levels of omentin-1 were measured using ELISA. The circadian secretion pattern was examined for an underlying periodicity using nonlinear four-parameter cosine OLS regression. The omentin-1 levels between the 3 conditions were compared using a Hierarchical Mixed-Effects Linear Modeling approach. The omentin-1 levels over time after bariatric surgery was examined using repeated measures ANOVA.[br]Results: Omentin-1 levels showed no circadian variation (p=0.92). Omentin-1 levels in the fed, fasting, and fasting+leptin states were 261, 208 and 331 ng/mL respectively (p=0.14). Mean (standard error of the mean, SEM) omentin-1 levels before bariatric surgery and after 3 and 6 months were 304 (28), 332 (36) and 273 (31) ng/mL respectively. Subgroup analysis by type of surgery did not find any change in omentin-1 levels in either type of surgery (laparoscopic gastric banding (p=0.65) or Roux-en-Y (p=0.63)).[br]Conclusions: There is no circadian variation in secretion of omentin-1, and levels of omentin-1 are not altered by acute or chronic energy deficiency, by type of bariatric surgery or by administration of leptin.[br][br](1) Yang RZ et al. Identification of omentin as a novel depot-specific adipokine in human adipose tissue: possible role in modulating insulin action. Am J Physiol Endocrinol Metab 2006;290:E1253-61. (2) Moreno-Navarrete JM et al. Circulating omentin concentration increases after weight loss. Nutr Metab (Lond) 2010;7:27.[br][br]Sources of Research Support: This study was supported by National Institutes of Health [ndash] National Center for Research Resources grant M01-RR-01032 (Harvard Clinical and Translational Science Center) and grant number UL1 RR025758. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Center for Research Resources or the National Institutes of Health. Funding was also received from the National Institute of Diabetes and Digestive and Kidney Diseases grants 58785, 79929 and 81913, and AG032030. Amylin Pharmaceuticals, Inc. supplied metreleptin for this study but had no role in the study design; conduct of the study; collection, management, analysis, and interpretation of the data; or the preparation, review, or approval of the manuscript.[br][br]Nothing to Disclose: O-PRH, BT, KNA, CSM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1006 295 2025 MON-90 PO27-03 Monday 1693 2012


1690 ENDO12L_MON-91 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Different Growth Hormone, Ghrelin and Peptide YY[sub]1-36[/sub] Secretion after Oral Glucose in Women and Men Elena Outeirino, Jesus Garcia-Buela, Lara Pena, Susana Sangiao-Alvarellos, Sonia Pertega-Diaz, Teresa Martinez-Ramonde, Fernando Cordido University Hospital A Coruna, A Coruna, Spain; University Hospital A Coruna, A Coruna, Spain; University Hospital A Coruna, A Coruna, Spain; University Hospital A Coruna, A Coruna, Spain; University of A Coruna, A Coruna, Spain Sexual dimorphism of GH secretion is unclear in the human. There is evidence that oral glucose (OG) administration initially decreases and subsequently stimulates GH secretion.[br][bold]Objective:[/bold] Our aim was to study fasting GH concentrations and their response to OG administration in obese and healthy women and men, in order to elucidate the mecanism of sexual dimorphism of GH secretion and the possible contribution of ghrelin.[br][bold]Participants and Methods:[/bold] We included obese and healthy subjects, 33 women and 11 men. After an overnight fast, 75g of oral glucose were administered; glucose, insulin, ghrelin and PYY[sub]1-36[/sub] were obtained at baseline and during 300 minutes.[br][bold]Results:[/bold] Fasting GH ([micro]g/l) was higher in women than men; 1.3[plusmn]0.3 vs 0.2[plusmn]0.1, p=0.009, for women and men respectively. The area under the curve (AUC) between 0 and 150 min of GH ([micro]g/l[middot]min) was higher in women than men; 98.2[plusmn]25.9 vs 41.5[plusmn]28.6, p=0.002, for women and men respectively. The AUC of total ghrelin (pg/ml[middot]min, mean[plusmn]SEM) between 0 and 150 min was borderline significantly higher in women than men; 128562.3[plusmn]8335.9 vs 98839.1[plusmn]7668.6, p=0.069, for women and men respectively. Several initial time points were higher in women than men. Glucose, insulin and PYY[sub]1-36 [/sub]was similar in women and men after OG. There was a positive correlation between the AUC of total ghrelin and GH between 0 and 150 min (p=0.011).[br][bold]Conclusions:[/bold] Fasting and initial GH secretion is higher in women than men, in contrast peak and late GH secretion is similar. Sexual dimorphism in the regulation of GH secretion probably involves ghrelin.[br][br]Nothing to Disclose: EO, JG-B, LP, SS-A, SP-D, TM-R, FC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 559 295 2026 MON-91 PO27-03 Monday 1694 2012


1691 ENDO12L_MON-92 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Changes in Appetite Regulatory Peptides Following Gastric Bypass in Adolescents Janet Chuang, Todd Jenkins, David D[apos] Alessio, Thomas Inge Cincinnati Children[apos]s Hospital Medical Center, Cincinnati, OH; Cincinnati Children[apos]s Hospital Medical Center, Cincinnati, OH; University of Cincinnati, Cincinnati, OH Background: Roux en Y gastric bypass (RYGB) is effective in producing prolonged and significant weight loss. One of the mechanisms likely responsible is the change in the secretion of gut peptides that regulate energy balance. Prior studies in adults demonstrated enhanced postprandial release of the satiety signaling hormones peptide YY (PYY) and glucagon like peptide-1 (GLP-1) and suppression of orexigenic ghrelin. RYGB is now being increasingly offered in severely obese adolescents. It is unknown how RYGB may affect key GI hormones related to energy balance in adolescents.[br]Objective: Our aim was to describe changes in serum GLP-1, PYY, and ghrelin concentrations in adolescents after undergoing RYGB.[br]Methods: 14 subjects (12-21 years of age) were recruited from a cohort of adolescents undergoing bariatric surgery at Cincinnati Children[apos]s Hospital. They were studied before and 2 weeks, 3 months, and 1 year post- RYGB. Pre- and post-prandial serum profiles of GLP-1, PYY, and ghrelin were measured during a 90 minute meal tolerance test (MTT, 474 kcal Pediasure).[br]Results: Of the 56 planned MTTs, 28 were completed. Nine subjects had a baseline MTT, 4 had a 2 week MTT, 6 had a 3 month MTT, and 9 had a 1 year MTT. One year following surgery, body mass index declined, on average, by 33%. GLP-1 profiles significantly differed over time (p[lt]0.01). Specifically, peak levels were greater at the 3 month and 1 year visits compared to baseline. Postprandial PYY secretion trajectories were similar over time; however, PYY values at each post-op assessment were significantly greater than at baseline (p[lt]0.05 for each). Postprandial ghrelin secretion was found to differ by study visit (p=0.04).[br]Conclusion: This pilot data in adolescents suggests that alterations in the regulation of the satiety signaling hormones may occur in adolescents following RYGB. The current data justify further detailed investigations with larger cohorts to examine how weight loss operations may affect energy balance pathways in adolescents.[br][br]Nothing to Disclose: JC, TJ, DD, TI 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1803 295 2027 MON-92 PO27-03 Monday 1695 2012


1692 ENDO12L_MON-93 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Decreased Adiponectin in Mexican-American Versus Non-Latino White Subjects Is Not Explained by Differences in Adiposity or Diet Rocio I Pereira University of Colorado Anschutz Medical Campus, Aurora, CO; Denver Health, Denver, CO Decreased adiponectin is associated with insulin resistance and predicts type 2 diabetes. We have previously reported decreased adiponectin in Mexican Americans (MA) compared to non-Latino white (W) patients with hypertension (1), suggesting a contribution of decreased adiponectin to higher incidence of diabetes in MA. However, environmental factors leading to ethnic disparities in adiponectin have not been identified. We hypothesized that ethnic differences in dietary habits are associated with decreased adiponectin in MA.[br]We measured adiponectin in otherwise healthy MA and W, lean, overweight, and obese, men and women. The two ethnic groups were matched for BMI and gender. Adiposity was measured by DEXA scan, and dietary habits were recorded using the Block Food Frequency questionnaire. Adiponectin was measured by RIA. The homeostatic model of insulin resistance HOMA2 IR measure was calculated from fasting glucose and insulin levels.[br]Subjects in the matched groups included 30 participants- 5 men and 10 women in each ethnic group (age 34[plusmn]7). The two groups were well matched for BMI (kg/m[sup]2[/sup], 28.3[plusmn]0.8 in MA vs. 27.4[plusmn]1.0 in W). There were no significant group differences in adiposity (% body fat, fat mass, FFM mass, waist circumference). Adiponectin was lower in MA compared to W (5.7[plusmn]0.9 vs. 10.1[plusmn]1.1 [micro]g/ml, p=0.005).[br]The MA group had a lower consumption of seed nuts (0.3 vs. 0.9 meat equivalents, p=0.03), a lower percentage of calorie intake from fat (32.6 vs. 35.1%, p=0.043), and lower percentage of calorie intake from alcohol (1.0 vs. 5.0%, p=0.006). There were no group differences in total caloric intake, protein, fat, or carbohydrate intake, or intake of fiber, fruit servings, or grain servings. In addition, there were no group differences in intake of dietary fats. Controlling for age, gender, and dietary differences, adiponectin remained significantly correlated to MA ethnicity (adjusted R[sup]2[/sup]=0.476, p=0.006). HOMA2 IR was higher in the MA group though the group difference did not reach statistical significance. Furthermore, adiponectin was not correlated with HOMA2 IR in our subject groups.[br]We conclude that adiponectin is decreased in healthy MA compared to W individuals. Neither differences in adiposity nor differences in dietary habits explained observed ethnic group differences in adiponectin. Additional research is needed to identify environmental factors contributing to decreased adiponectin in MA populations.[br][br](1)Pereira RI et al., BMC Endocrine Disorders 2011; 11:13.[br][br]Sources of Research Support: NIH/NCRR Colorado CTSI UL1 RR025780; NIH/NCRR K23 RR22238.[br][br]Nothing to Disclose: RIP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1871 295 2028 MON-93 PO27-03 Monday 1696 2012


1693 ENDO12L_MON-94 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Human Memory CD4 T Cell Activation and HIV-1 Production Are Enhanced by Primary Human Subcutaneous Preadipocytes, Mediated in Part by IL-2 and IL-6 Jacob Couturier, Ashok Balasubramanyam, Dorothy E Lewis University of Texas Health Science Center at Houston, Houston, TX; Baylor College of Medicine, Houston, TX T cell and macrophage infiltration into adipose tissue is elevated during HIV infection, as it is in metabolic diseases such as obesity, insulin resistance, and diabetes. A major proportion of adipose tissue-resident T cells are memory CD4 T cells, which are the primary hosts for HIV infection, as well as the main cellular reservoir during latent infection. Many of these memory CD4 T cells within adipose tissue are in an activated state, suggesting that adipose tissue is a stimulatory environment conducive to promoting HIV replication. As both memory CD4 T cell activation and HIV replication are highly susceptible to regulation by proinflammatory cytokines and chemokines, including those produced by preadipocytes and mature adipocytes (e.g., TNF[alpha], IL-6, RANTES, MCP-1), we tested the hypothesis that memory CD4 T cell activation and HIV replication could be enhanced by preadipocytes. Short-term transwell (no contact) co-culture experiments were performed between in vitro HIV-infected, resting memory CD4 T cells (purified from peripheral blood) and primary human subcutaneous preadipocytes. Flow cytometry staining for T cell activation markers such as CD69 and CD25 showed that the preadipocytes robustly activated memory CD4+ T cells by three days of co-culture, in a manner dependent on the presence of IL-2. Blocking antibodies against IL-6 (in the presence of IL-2) mitigated CD69 expression by approximately one-third, indicating a combinatorial role for IL-6 and IL-2 in T cell activation by preadipocytes. Blocking antibodies against TNF[alpha] did not reduce memory CD4+ T cell activation by preadipocytes, nor did blocking antibodies against T cell chemokine receptors (CCR5, CXCR4, or CXCR3) or their respective ligands (RANTES, MIP-1, MCP-1, or SDF-1). Lastly, production of primary strains of HIV (as measured by intracellular and extracellular p24) was enhanced by preadipocytes concomintant with T cell activation, and was also mediated by IL-2 and IL-6. As preadipocytes are principal producers of IL-6 in adipose tissue, these findings suggest that adipose tissue preadipocytes are important regulators of T cell activation and HIV replication within adipose tissue.[br][br]Nothing to Disclose: JC, AB, DEL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1952 295 2029 MON-94 PO27-03 Monday 1697 2012


1694 ENDO12L_MON-95 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Endotoxin Reveals Human Adipose Genes Suppressed by Obesity [italic]In Vivo[/italic] Rachana Shah, Christine C Hinkle, Nehal N Mehta, Mary Putt, Muredach P Reilly Children[apos]s Hospital of Philadelphia, Philadelphia, PA; University of Pennsylvania, Philadelphia, PA; University of Pennsylvania, Philadelphia, PA Adipose inflammation is a crucial link between obesity and its related metabolic complications. Human experimental endotoxemia is a controlled model for the study of inflammatory responses [italic]in vivo[/italic]. We hypothesized that adipose gene expression during endotoxemia would approximate the changes observed with obesity-related adipose inflammation and reveal candidates involved in metabolic processes. Utilizing microarray analysis of human subcutaneous adipose tissue before and during endotoxemia we identified genes suppressed by evoked inflammation and validated the expression changes in human obesity.[br]Healthy volunteers (n=14) were subject to a high-dose (3 ng/kg) endotoxin challenge; adipose biopsies were taken at 0, 4, 12, and 24 hours for microarray analysis. A priority list of 68 downregulated genes ([lt]67% of baseline expression) was validated by RT-PCR in an independent sample of adipose tissue from healthy subjects (n=7) after a more physiologically relevant low-dose (0.6 ng/kg) endotoxin challenge. Expression of validated genes was screened in adipose tissue of lean and obese humans (n=11/group), and the cellular source of the genes was probed in cultured human adipocytes and monocyte/macrophages.[br]Endotoxin (3 ng/kg) resulted in suppression of expression in 353 unique genes. In the low-dose endotoxin validation, 22 (32%) of the 68 prioritized genes were downregulated. Of the validated genes, 50% (11/22) were downregulated in primary human adipocytes after treatment with endotoxin, and 32% (7/22) showed decreased expression in macrophages after polarization to the inflammatory M1 phenotype, leaving 4/22 (18%) with likely other cellular source. Comparing expression in obese vs lean human adipose, 50% (11/22) were confirmed to have decreased expression ([lt]50% of lean) in obese human subjects (p[lt]0.05). Functional classification of these genes reveals involvement in transcriptional regulation, endocrine signaling, cell adhesion, molecular clock, and immune regulation.[br]Exploration of gene expression changes in adipose tissue during human endotoxemia revealed suppression of many genes with potential implications in obesity and metabolic dysfunction. The cellular source of these genes includes adipocytes, infiltrating macrophages, and other adipose tissue cells. A majority of the candidates were also suppressed in endogenous human obesity, suggesting a biologic role in the pathophysiology of obesity-related adipose inflammation.[br][br]Nothing to Disclose: RS, CCH, NNM, MP, MPR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1968 295 2030 MON-95 PO27-03 Monday 1698 2012


1695 ENDO12L_MON-96 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) TRAIL (TNF-Related Apoptosis-Inducing Ligand) Regulates Human Adipocyte Metabolism by Caspase-Mediated Cleavage of PPAR[gamma] Pamela Fischer-Posovszky, Michaela Keuper, Ingrid Wernstedt Asterholm, Philipp E Scherer, Klaus-Michael Debatin, Martin Wabitsch Ulm University Medical Center, Ulm, Germany; University of Texas Southwestern Medical Center, Dallas, TX; Ulm University Medical Center, Ulm, Germany [bold] Background:[/bold][br]TNFalpha and other members of the tumor necrosis factor family affect adipose tissue metabolism and contribute to the obesity-related inflammation of adipose tissue. Here, we sought to identify the effects of TRAIL (TNF related apoptosis inducing ligand) on human fat cell biology.[br][bold] Methods: [/bold] SGBS adipocytes were used as an [italic] in vitro [/italic] model for human adipocyte biology. Expression of TRAIL and TRAIL receptors was studied in adipose tissue of fasted and re-fed mice and also in human adipose tissue samples.[br][bold] Results: [/bold] In SGBS adipocytes TRAIL inhibited insulin-stimulated glucose uptake in a dose dependent manner ([sim] 45 % at TRAIL 100 ng/ml and 10 nM insulin). Insulin-stimulated [italic]de novo[/italic] lipogenesis was inhibited to a comparable extent ([sim] 50 % at TRAIL 100 ng/ml and 10 nM insulin). By binding to TRAIL-receptor 2 (TRAIL-R2), TRAIL activates cleavage of caspase-8 and -3, which in turn cleaves and inactivates PPARgamma resulting in reduced expression of lipogenic genes such as Glut-4, FASN, and ACC. Most interestingly, TRAIL-R2 and its mouse homologue DR5 are regulated upon acute and chronic energy imbalance in murine and human adipose tissue.[br][bold] Summary and Conclusions: [/bold] Taken together we discovered a so far unknown function of the death ligand TRAIL in regulating human adipocyte metabolism. Our results imply that blocking TRAIL signaling in adipocytes might provide a new strategy to prevent obesity-induced insulin resistance. Furthermore, our findings have important implications for cancer research since agonistic TRAIL receptor antibodies are currently tested as anti-cancer agents.[br][br]Sources of Research Support: PFP receives a Margarete von Wrangell fellowship financed by the Ministry of Science, Research and the Arts in Baden-Wuerttemberg and the European Social Fund.[br][br]Nothing to Disclose: PF-P, MK, IWA, PES, K-MD, MW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1485 295 2031 MON-96 PO27-03 Monday 1699 2012


1696 ENDO12L_MON-97 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Basal and Stimulated Leptin Secretion in African Americans and Caucasians Chimaroke Edeoga, Sotonte Ebenibo, Samuel Dagogo-Jack University of Tennessee Health Science Center, Memphis, TN Background: Leptin regulates body weight, and ethnic disparities in circulating levels have been suggested. However, definitive studies are lacking. Also, responses to leptin secretagogues have not been well characterized.[br]Subjects and Methods: We studied 292 (137 white, 155 African American {AA}) nondiabetic subjects (age 18-65 years) in our Pathobiology of Prediabetes in a Bi-racial Cohort (POP-ABC) study[sup]1[/sup]. Baseline assessments in POP-ABC included physical exam, OGTT, body fat (DEXA), fasting leptin, insulin, cortisol, lipids, and other analytes. Dynamic leptin secretion was assessed using the method of Dagogo-Jack et al,[sup] 2[/sup] with a reduced dose (2 mg) of dexamethasone (dex). Leptin data were compared across gender and ethnicity, and interactions with body fat and other variables were analyzed using ANOVA and linear regression.[br]Results: Among women, AA had higher mean ([plusmn]SEM) fasting leptin (50.6 [plusmn] 3.33 vs. 39.5 [plusmn] 3.51 ng/ml, P=0.001), but male values were similar in AA and whites(12.4 [plusmn] 2.06 vs. 11.1 [plusmn] 1.4 ng/ml). BMI and total fat mass also were higher in AA women than whites. However, the relation between fasting leptin and total fat was similar in AA and whites (r=0.81, P [lt]0.0001). Peak leptin response to dex (% baseline) was 227 [plusmn] 11.6 % in AA and 205 [plusmn] 6.6% in whites (P=0.065), and similar in men vs women (217 [plusmn] 9.1% vs. 204 [plusmn] 6.7%). The delta leptin responses were higher in AA men than whites (P=0.05). From sensitivity analyses 29 Subjects (15AA, 14 whites) were [gt]90[sup]th[/sup] percentile and 74 subjects (38 AA, 36 whites) were [lt]25[sup]th[/sup] percentile responders to leptin secretagogue. Univariate analysis showed interactions between delta leptin response vs. fasting leptin ([lt]0.0001), total fat mass (P[lt]0.0001), BMI (P[lt]0.0001), HOMA-B (P=0.0027), HOMA-IR(P[lt]0.0001), triglycerides (P=0.0078), fasting glucose (P=0.027), CRP (P=0.027) and systolic blood pressure (P=0.037). Age, race and gender did not predict leptin responses. In a multivariate model, only fasting leptin emerged as a significant predictor (r= -0.26, P=0.037).[br]Conclusion: Our data do not indicate significant ethnic differences in basal leptin secretion in relation to fat mass. Also, dynamic leptin secretion in response to secretagogues did not differ by gender, but were modestly greater in AA men than whites. Dynamic leptin secretory capacity is inversely related to baseline leptin, glucose, insulin, systolic BP, triglycerides and adiposity, and positively related to insulin sensitivity.[br][br]1Dagogo-Jack S, et al. Pathobiology of Prediabetes in a Biracial Cohort (POP-ABC): Design and Methods. Ethn Dis 21:33[ndash]39, 2011. 2Dagogo-Jack S, et al. Robust leptin secretory responses to dexamethasone in obese subjects. J Clin Endocrinol Metab 82:3230-3233, 1997.[br][br]Sources of Research Support: Grants from the NIH(R01 DK067269 and MO1 RR00211) and the American Diabetes Association.[br][br]Nothing to Disclose: CE, SE, SD-J 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2298 295 2032 MON-97 PO27-03 Monday 1700 2012


1697 ENDO12L_MON-98 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Real-Time Redox Response of Human Preadipocytes to Fatty Acids Carlyle Rogers, Barbara Davis, Jacques Robidoux East Carolina University Brody School of Medicine, Greenville, NC Failure of adipose tissue to adapt to chronic positive energy imbalances may cause or expedite the onset of systemic insulin resistance due to fatty acid (FA) efflux to extra-adipose organs. Although high FA levels cause lipotoxicity in cell types involved in fuel homeostasis, there is an ingrained notion that preadipocytes are immune to this response. As this notion was challenged in rodent models[italic], [/italic]we quantified the response of human preadipocytes to a physiological mixture of FA (oleate, palmitate, stearate, linoleate) within the physiological FA to albumin ratio. We chose preadipocytes because their loss could cause a failure in adipose tissue storage capacity.[br]Since our preliminary data suggested that insulin-resistant individuals have a reduced preadipocyte antioxidant capacity, we tested the impact of FA on their redox status. Experiments using our novel continuous cytometry method showed that the maximal rate of mitochondrial superoxide accumulation occurred within seconds after FA exposure. However, the mitochondrial reactive oxygen species (ROS) handling system was able to buffer this surge as no increase in cytosolic H[sub]2[/sub]O[sub]2[/sub] was measured within 12 hrs. Nevertheless, FA at concentrations above 600 uM led to an increase in cytosolic H[sub]2[/sub]O[sub]2[/sub] from 18 hrs, which ultimately led to the death of 35% of cells. These events were prevented by the use of mitochondria selective scavengers of superoxide and H[sub]2[/sub]O[sub]2[/sub].[br]Although no overt oxidative stress occurred at either of our low concentrations or within 12 hours, we still observed the activation of the prototypical sirtuin 1-forkhead box O (FOXO) stress-dependent pathway as early as 3 hours post exposure. Activation of this pathway was, like the toxic effects, prevented by blocking mitochondrial ROS accumulation. Interestingly, inhibition of sirtuins exacerbated the toxic effects of FA. Altogether, this study suggests that activation of a mitochondria-FOXO pathway could protect preadipocytes from lipotoxicity and prevent a reduction in the preadipocyte pool.[br][br]Sources of Research Support: American Heart Association Mid-Atlantic Affiliate Predoctoral Fellowship; East Carolina Metabolic and Obesity Institute.[br][br]Nothing to Disclose: CR, BD, JR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 211 295 2033 MON-98 PO27-03 Monday 1701 2012


1698 ENDO12L_MON-99 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) High Glucose (HG) Induced Inflammation and Intracellular Lipid Accumulation in Human Bone Marrow-Derived Mesenchymal Stem Cells (MSCs) Is Reduced by Mitochondrial Antioxidant Manganese Superoxide Dismutase (MnSOD) Up-Regulation Sabyasachi Sen, Cyril Chou, Mary Young, Young-Cheul Kim, Nagendra Yadava Baystate Medical Center, Springfield, MA; Pioneer Valley Life Science Institute, Springfield, MA; University of Massachusetts, Amherst, MA [bold]Background: [/bold]Primary MSCs can differentiate into mature adipocytes, osteocytes or chondrocytes. Behavior of MSCs in HG concentrations is largely unknown. In HG, we noted increased intracellular lipid accumulation (confirmed by Oil Red O stain). We studied gene expression (qRT-PCR) of HG exposed MSCs. Based on our observations we postulated that HG is associated with increased reactive oxygen species (ROS) accumulation particularly in mitochondria that promoted inflammation and adipogenic differentiation of MSCs. Therefore, intra-cellular antioxidant upregulation could counteract the ROS accumulation, reduce associated inflammation and adipogenic differentiation of MSC in HG. [bold]Methods: [/bold]We exposed MSCs to HG (25mM) and normal glucose (NG, 5.5 mM) and Oil Red O positivity with changes in gene expression were noted. ROS generation was analyzed in cytosol and mitochondria using DCF-DA and Mitosox-Red stain, respectively. We interrogated mitochondrial respiration (Seahorse) and complexes 1 [amp] 2 protein by BN-PAGE and SDS-PAGE. Next we transduced hMSCs with Adenovirus containing MnSOD, CAT or control gene GFP (green florescent protein) genes at 100 MOI, to over-express these specific genes prior to HG exposure. In-vitro co-culture of adipocytes and transduced MSCs (3:1 ratio) were set up in NG and HG. [bold]Results: [/bold]HG increased lipid accumulation (3.5 fold), increased ROS accumulation in both cytosol and mitochondria. HG increased adipogenic gene expression of Leptin (10-fold), Perilipin (4-fold), PPARG (16 fold) while it reduced bone formation markers Alkaline Phosphatase (4 fold), osteocalcin (1.6 fold) and osteopontin (4 fold) mRNAs. HG increased TNF[alpha] (Tumor Necrosis Factor), IL6 (interleukin6) and Endothelin1, inflammatory marker gene expressions, significantly. Mitochondrial complex analyses indicated impaired mitochondrial respiration and suppressed complex 1 protein expression in HG exposed MSCs. MnSOD much more than CAT up-regulation, reduced adipogenic and inflammatory gene expressions and rescued suppressed Complex-1 and reduced HG associated mitochondrial ROS accumulation (Mitosox-Red). Co-culture of mature adipocytes and MnSOD upregulated MSC reduced lipid droplets in adipocytes. [bold]Conclusions: [/bold]Upregulation of mitochondrial anti-oxidant (MnSOD[gt]CAT) effectively reduced inflammation, adipogenic differentiation markers and improved mitochondrial respiration in HG. To address HG associated obesity, in-vivo studies using MnSOD upregulated MSCs is being conducted.[br][br]Sources of Research Support: Collaborative Biomedical Research Grant.[br][br]Nothing to Disclose: SS, CC, MY, Y-CK, NY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1284 295 2034 MON-99 PO27-03 Monday 1702 2012


1699 ENDO12L_MON-100 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) The Effects of Metformin on Cultured Human Mammary Adipose Tissue Sully Fernandez, Paul A Volden, Maxwell N Skor, Feenalie Patel, Julie Park, Matthew J Brady, Suzanne D Conzen University of Chicago, Chicago, IL; University of Chicago, Chicago, IL The exponential increase in obesity in the United States is a major public health concern. In addition to an enhanced risk for type 2 diabetes and cardiovascular disorders, human obesity is associated with increased rates of certain cancers including breast cancer. Recently, the anti-diabetic drug metformin was shown to have a potential role in breast cancer prevention. In this study, we have begun to investigate the contribution of obesity to a pro-tumorigenic local mammary adipose tissue microenvironment and the potential reversal of this effect by metformin. Otherwise-discarded human breast adipose tissue was collected from five patients undergoing elective reduction mammoplasty. Following resection, the adipose tissue was immediately minced and placed in 10% FBS-containing DMEM in the absence or presence of 10mM metformin for 24 hours. Tissue was then washed thoroughly and cultured in serum-free media with 0.1% BSA for 8 hr. The resulting culture media was collected for eventual analysis of secreted adipokines. The adipose tissue was then rinsed thoroughly and incubated with varying concentrations of insulin (0-50 nM) for 10 minutes. Adipocytes were lysed by sonication in protein homogenization buffer, centrifuged to remove debris and nuclei, and stored at -80oC. Batched lysates were analyzed by Western immunoblotting using the following antibodies: Anti- phospho-specific Akt Ser 473 and total-Akt, anti- phospho-specific Thr 172 AMP-kinase (AMPK) and total AMPK, and anti- phospho-specific Ser211 glucocorticoid receptor (GR) and total GR. In these preliminary studies, high patient BMI ([gt]30) was associated with enhanced basal phosphorylation of AMPK, while in leaner subjects, pAMPK was low at baseline and metformin resulted in increased AMPK phosphorylation. Interestingly, in all patient samples (regardless of BMI), pre-incubation of the primary adipose tissue with metformin resulted in significantly reduced insulin-stimulated phosphorylation of Akt at Ser473. These results suggest that, in addition to a directly antagonistic effect of metformin on human breast cancer cell proliferation, metformin may also have a previously unidentified role in affecting the mammary adipose tissue microenvironment. The effects of metformin-induced changes on mammary adipose tissue biology could play an important role in breast cancer biology and deserves further investigation.[br][br]Sources of Research Support: DOD Fellowship W81XWH-11-1-014901;NIH Grant R01CA148814.[br][br]Nothing to Disclose: SF, PAV, MNS, FP, JP, MJB, SDC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2055 295 2035 MON-100 PO27-03 Monday 1703 2012


1700 ENDO12L_MON-101 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Macrophage Removal Increases Human Preadipocyte Differentiation in Omental Adipose Tissue Li Fen Liu, John M Morton, Lorna L Tolentino, Sam Cushman, Edgar G Engleman, Tracey L McLaughlin Stanford University, Palo Alto, CA; National Institutes of Health, Bethesda, MD [bold]Objective:[/bold] Macrophage infiltration in human adipose tissues is associated with obesity and insulin resistance. Impaired adipocyte differentiation may also contribute to insulin resistance, but factors determining adipogenic potential are unclear. The aim of the current study was to investigate whether macrophages in play a role in depot-dependent differences in adipocyte differentiation by culturing human preadipocytes in the presence or absence of[bold] CD45+ cells.[/bold][br][bold]Methods:[/bold] Abdominal subcutaneous and omental adipose tissues were obtained from nine subjects undergoing bariatric surgery. After isolation of stromalvascular cells (SVCs), immune cells including macrophages (CD45-positive cells) were removed by immune magnetic separation in omental visceral adipose tissues. Preadipocytes isolated from subcutaneous adipose tissues (SAT), CD45-positive and negative (CD45 +, CD45-) omental adipose tissues were differentiated into adipocytes [italic]in vitro[/italic] for 14 days. Adipogenesis was determined by measuring the number of differentiated adipocytes by immunostaining, Oil-Red O extraction and gene expression by qRT-PCR.[br][bold]Results:[/bold] Adipogenesis as determined by Oil Red extraction (SAT 78 [plusmn]5.6 vs. omental 6.7 [plusmn]2.1) and the percentage of differentiated adipocytes (SAT 60.0 [plusmn]14.7 vs. omental 13.9 [plusmn]7.3) were significantly lower (p[lt]0.001) in omental adipose tissue. Macrophage removal significantly improved the percentage of adipocyte differentiation in CD45- preadipocytes in omental adipose tissue (29.8 [plusmn]13.23,p[lt]0.001)). These changes in differentiation were associated with increased expression of adiponectin, a marker of terminal differentiation (0.06 [plusmn]0.07 to 0.35[plusmn]0.21, p=0.05) and decreased IL6, marker of inflammation, (0.85 [plusmn]0.22 to 0.017 [plusmn]0.15 p[lt]0.001) in CD45- omental adipose cells. PPAR[gamma] and CEBP[beta], markers of early adipocyte differentiation, were not significantly altered in omental adipose tissue after CD45 depletion.[br][bold]Conclusions: [/bold]Our data demonstrate a clear depot-dependent difference in human preadipocyte dfferentiation. The mechanism of this effect is unclear. However, given that macrophage removal improved adipocyte differentiation and increased adiponectin expression, it is plausible that inflammation impairs terminal differentiation and fat uptake in obese humans.[br][br]Nothing to Disclose: LFL, JMM, LLT, SC, EGE, TLM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2239 295 2036 MON-101 PO27-03 Monday 1704 2012


1701 ENDO12L_MON-102 POSTER SESSION: Human Adipose Biology [amp] Weight Regulation (1:30 PM-3:30 PM) Association of Insulin Resistance and Glp-2 Secretion in Obesity: A Pilot Study Daniela Tezoto, Bruno Geloneze, Marcelo Miranda Lima, Jose Carlos Pareja, Claudio Rodrigues Saddy Coy, Antonio Ramos Calixto, Sylka Rodovalho Geloneze, Daniela Oliveira Magro University of Campinas, Campinas, Brazil; University of Campinas, Campinas, Brazil [bold]Objective: [/bold]Glugagon like peptide 2 (GLP-2) is a potent intestinotrophic hormone. Our hypothesis is that insulin sensitivity is influenced by increased absorption of nutrients mediated this hormone. The objective of this pilot study was to determine whether GLP-2 secretion relates to insulin sensitivity (IS) in obese subjects.[br][bold]Methods: [/bold]Twenty four obese subjects [body mass index (BMI) 40.0[plusmn]3.0 Kg/m[sup]2[/sup] (mean[plusmn]standard deviation)] were included, nine of which were male, age 43[plusmn]8 years. Twelve subjects had type 2 diabetes, all treated with oral anti-diabetic agents only. The subjects were submitted to standard meal tolerance test (MTT) for dosage of the curves: glucose, insulin and GLP-2. The insulin sensitivity was measured by HOMA-IR and OGIS derived from the MTT. Spearman linear correlations and partial correlations were obtained.[br][bold]Results: [/bold]There was an inverse relationship between the GLP-2 secretion and IS: HOMA-IR correlated with GLP-2 AUC (R=0.504; p=0.012) and OGIS correlated with GLP-2 incremental AUC (R=-0.54; p=0.054). The correlation persisted controlling for BMI.[br][bold]Conclusion: [/bold]We found an association of GLP-2 secretion and insulin resistance. The comprehension of the underlying mechanisms may provide future directions in the pharmacological manipulation of incretins and in the treatment of obesity and related metabolic disorders.[br][br]Sources of Research Support: The present study was funded by FAPESP, Sao Paulo, Brazil (Protocol n. 2009/50430-6).[br][br]Nothing to Disclose: DT, BG, MML, JCP, CRC, ARC, SRG, DOM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1645 295 2037 MON-102 PO27-03 Monday 1705 2012


1702 ENDO12L_MON-103 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Insulin Secretion in Mice Deficient in the Slc12a2 Gene Saeed Alshahrani, Mauricio Di Fulvio Wright State University - Boonshoft School of Medicine, Dayton, OH Insulin secretion from pancreatic [beta]-cells is a complex process involving the integration and interaction of multiple external and internal stimuli. The most well known mechanism whereby glucose induces insulin secretion involves the ATP-dependent K+-channel pathway. Glucose metabolism within the [beta]-cell increases the ATP/ADP ratio closing K+-channels and thus depolarizing the plasma membrane. This activates voltage-dependent Ca2+-channels inducing Ca2+ influx and triggering insulin secretion from stored granules. However, in addition to this cationic pathway, glucose also stimulates Cl[ndash] efflux. This contributes to [beta]-cell[apos]s electrical and secretory activities. The electrogenic efflux of Cl[ndash] in response to glucose is possible because [beta]-cells maintain the intracellular chloride concentration ([Cl[ndash]]i) above electrochemical equilibrium due to the functional presence in these cells of the Na+K+2Cl[ndash] co-transporter, a protein encoded by the Slc12a2 gene. This co-tranporter is the target of loop-diuretics such as bumetanide or furosemide. Accordingly, these drugs are known to impair insulin secretion in response to glucose. However, our results show that glucose-induced insulin secretion is far from being impaired in mice carrying heterozygous or homozygous inactivation of the Slc12a2 gene. Transgenic mice lacking one or both functional alleles of this gene have better fasting glycemia and improved glucose tolerance when compared to WT. Further, mice deficient in the Slc12a2 gene are more sensitive to glucose when compared to WT littermates whereas overall insulin sensitivity is maintained in these mice, regardless of genotype. Together, or results suggest for the first time that i) the Slc12a2 gene is not required for glucose-induced insulin secretion, ii) additional mechanisms are involved in regulating [Cl[ndash]]i in [beta]-cells and, iii) hemiexpression of the Slc12a2 gene in these mice may have a positive impact in glucose homeostasis.[br][br]Sources of Research Support: This work was supported by funds awarded to MDiF by the American Diabetes Association (Grant 1-10-JF-14).[br][br]Nothing to Disclose: SA, MDF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 433 296 2038 MON-103 PO13-01 Monday 1706 2012


1703 ENDO12L_MON-104 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Pancreatic Deletion of 12-Lipoxygenase Protects Against Diet-Induced Glucose Intolerance Sarah A Tersey, Jerry L Nadler, Raghavendra G Mirmira Indiana University School of Medicine, Indianapokus, IN; Eastern Virginia Medical School, Norfolk, VA There is accumulating evidence that high fat diets (HFDs) lead to both obesity and insulin resistance. In this setting, pancreatic islets hypersecrete insulin during a morphologic process known as adaptive hyperplasia. It is becoming clear that progressive defects in insulin secretion and/or adaptive hyperplasia drive the transition from normoglycemia to overt diabetes mellitus. Proinflammatory cytokines released by dysfunctional adipose tissues and macrophages in the setting of HFD and obesity are thought to signal to the islet [beta] cell, activating intracellular pathways that impinge upon insulin production and secretion. 12-lipoxygenase (12-LO) catalyzes the oxygenation of cellular poly-unsaturated fatty acids to form lipid inflammatory mediators, mainly 12-HETE. 12-LO(LoxP/LoxP) mice were crossed with Pdx-Cre mice to create a pancreas specific deletion of 12-LO (pLOKO) starting at the inception of pancreas development. pLOKO mice exhibited no differences in weight, growth, or glucose tolerance. pLOKO and their wildtype (WT) counterparts were then fed a normal chow (NC) or a high fat diet (HFD, 42% calories from fat) for 12 weeks to induce insulin resistance and glucose intolerance. pLOKO mice placed on NC had normal glucose and insulin tolerance, identical to their WT counterparts. Body weight gain was similar between WT and pLOKO mice on NC. Following HFD feeding, pLOKO mice gained weight similarly to WT mice. Dual-energy X-ray absorptiometry showed that body fat distribution was indistinguishable between WT and pLOKO mice on a HFD. Moreover, pLOKO mice on a HFD showed similar insulin resistance to WT mice on a HFD. Strikingly, however, pLOKO mice fed a HFD showed glucose tolerance that was significantly better than WT mice on the same diet, and comparable to mice on NC. Islets from pLOKO mice and their WT counterparts were incubated with and without a mixture of cytokines (IL-1beta, TNF-alpha, and IFN-gamma) for 18 hours and glucose-induced insulin-secretion was measured. Glucose-stimulated insulin secretion in islets isolated from pLOKO mice remained protected from cytokine-induced dysfunction after 18 hours of incubation with cytokines. Taken together, these data suggest that 12-LO and its products may play a central role in metabolic stress-induced inflammatory dysfunction in islet beta cells, and that inhibition of 12-LO may serve as a viable approach to preserve islet beta cell function in type 2 diabetes.[br][br]Nothing to Disclose: SAT, JLN, RGM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1634 296 2039 MON-104 PO13-01 Monday 1707 2012


1704 ENDO12L_MON-105 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Treating Diabetes with an Implanted Bioartificial Pancreas: Efficacy Depends on the Biomaterial Nicholas E Simpson, Michelle M Corrado, Srujana Neelam, Ansuya Deosaran, Mark J Beveridge University of Florida, Gainesville, FL Introduction: Type 1 diabetes (T1D) results from an autoimmune destruction of the insulin producing pancreatic beta-cells. An approach to treat T1D uses an implantable device combining cellular components with a non-biologic matrix (e.g., alginate) to take advantage of the cells[apos] ability to sense glucose and secrete insulin according to need; a bioartificial pancreas. This approach allows for restoration of metabolic homeostasis while potentially avoids the need for chronic immunosuppression. We previously demonstrated that alginate composition affects cell growth. We present here the therapeutic consequence of the cell-entrapping biomaterial of a bioartificial pancreas.[br]Methods: Female C3H/heN mice were rendered diabetic by one i.v. injection of alloxan (62.5 or 75 mg/kg), and body mass and blood glucose levels were monitored. After establishment of diabetes (blood glucose [gt]300 mg/dl), the bioartificial pancreas was implanted in the peritoneal cavity of the mice. The bioartificial pancreas consisted of TC-tet cells entrapped in one of two forms of gelled alginate (one alginate of high mannuronic acid content, LVM; one with high guluronic acid content, LVG) in a macroconstruct disc. Fasting glucose levels and body masses were monitored for one month. Blood insulin levels at the end of the study were measured and statistical tests between experimental groups and controls were performed.[br]Results: Alloxan induced T1D. Untreated animals all died rapidly, with a dose-dependent time to death (62.5 mg/kg all dead by day 15; 75 mg/kg all dead by day 5). Animals receiving the highest alloxan dose, and treated with the bioartificial pancreas all survived (n=16). The blood glucose of those receiving the LVG construct (n=9) maintained elevated ([gt]400 mg/dl) glucose levels. Those receiving the construct made with LVM (n=7) all had fasting blood glucose levels return to the normal glycemic range. Insulin levels measured at the conclusion of the study indicated that LVM construct recipients differed significantly from LVG-recipients (p[lt]0.001) but were similar to normal controls (p[gt]0.23); LVG recipient insulin levels were similar to diabetic mice (p[gt]0.76).[br]Conclusions: The choice of entrapping biomaterial has a profound effect on the function of the implanted bioartificial pancreas. This study demonstrates clear in vivo efficacy of a bioartificial pancreas in recipient diabetic animals (here a syngeneic model), provided an appropriate entrapping material is used.[br][br]Sources of Research Support: NIH NIDDK DK47858.[br][br]Nothing to Disclose: NES, MMC, SN, AD, MJB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1612 296 2040 MON-105 PO13-01 Monday 1708 2012


1705 ENDO12L_MON-106 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) miR-24-1 Targets MEN1 To Enhance Beta Cell Expansion in Pancreatic Islets Jyothi Vijayaraghavan, Elaine C Maggi, Judy S Crabtree Louisiana State University Health Sciences Center, New Orleans, LA In recent years it has been estimated that the global prevalence of type 2 diabetes (T2D) is approximately 150 million with projections suggesting that the number may double by 2025. T2D results due to peripheral insulin resistance and inadequate pancreatic beta cell mass to compensate for the increased physiological demand. In non-diabetic states, pancreatic islets can expand to enhance insulin secretion through various pathways to meet increased metabolic demand. One such pathway involves the reduction of menin levels during pregnancy or obesity resulting in beta-cell expansion via its effects on the cell cycle inhibitors p18ink4c and p27kip1(1,2). Elevated prolactin levels are responsible for reduced menin levels during pregnancy, but the same is not true for obesity where prolactin levels remain essentially unchanged. This suggests that other mechanisms exist to control menin levels and we propose that microRNAs (miRNAs) may contribute to the process of adaptive beta cell expansion in obesity. miRNAs are small, non-coding RNAs that regulate the expression of their mRNA targets by degrading them and/or inhibiting their translation. One such miRNA is miR-24-1 that has been observed to be upregulated in pancreatic insulinoma cells and has a seed sequence with homology to the 3[apos]UTR of MEN1. Here, we show that miR-24-1 interacts with the 3[apos] UTR of MEN1 and that the presence of miR-24-1 suppresses the levels of menin in insulinoma cells. Furthermore, beta cell expansion is an early phenotype in mice lacking menin in the pancreatic islets, and the lack of menin causes a significant decrease in miR-24-1 expression(3). This suggests that miR-24-1 could regulate beta cell expansion in the pancreatic islets by controlling the availability of menin, and that menin may impact the production of miR-24-1 via a negative feedback mechanism. Further, miR-24-1 may also be an important regulator of T2D pathogenesis because in addition to its proposed role in regulating menin, miR-24-1 has also been implicated in peripheral insulin resistance through its targeting of p38(4,5). We believe our findings will expand our understanding of the underlying mechanisms of diabetes pathogenesis and reveal novel therapies for its treatment.[br][br](1)Karnik et al., Science 2007; 318:806-9. (2)Karnik et al., Proc Natl Acad Sci U S A. 2005;102:14659-64. (3)Crabtree et al., Proc Natl Acad Sci U S A. 2001;98:1118-23. (4)Keren et al., Mol Cell Endocrinol 2006;252:224-30. (5)Thai et al., J Biol Chem 1998;273:14285-92.[br][br]Nothing to Disclose: JV, ECM, JSC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1005 296 2041 MON-106 PO13-01 Monday 1709 2012


1706 ENDO12L_MON-107 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Glucose Intolerance in Diet-Induced Obese, Adult-Onset Isolated GH-Deficient Mice Cannot Be Explained by Alterations in [beta]-Cell Mass Jose Cordoba-Chacon, Manuel D Gahete, Naveen Pokala, Raul M Luque, Rhonda D Kineman University of Illinois at Chicago, Chicago, IL; Jesse Brown Veterans Affairs Medical Center, Chicago, IL; University of Cordoba, Cordoba, Spain Mouse models of developmental GH deficiency (GHD) or GH resistance are dwarf and obese but have low circulating insulin levels, which may be due in part to improved systemic insulin sensitivity. However, mice with global or [beta]-cell specific knockout of the GH receptor (GHR) become glucose intolerant in response to high-fat (HF) feeding which is associated with impaired [beta]-cell expansion, suggesting GH also serves a direct role in maintaining [beta]-cell function/proliferation (1,2). Since circulating GH levels fall with age and weight gain, our laboratory has recently examined the metabolic impact of adult-onset isolated GHD (AOiGHD) in mice. We have previously reported that mean body weight of AOiGHD mice did not differ dramatically from controls. However, like developmental models of GHD and GH resistance, AOiGHD mice show improved insulin sensitivity and low circulating insulin levels, despite an increase in fat/lean mass (3). Also like the developmental dwarf models, AOiGHD mice become glucose intolerant in response to HF feeding, where insulin levels remain inappropriately low, as compared to obese controls. In the current study we investigated if the impaired insulin response observed in HF-fed AOiGHD mice was due to impaired function and/or expansion of the [beta]-cell population. We observed that expression of INS2, the primary insulin transcript in mice, was reduced in whole pancreatic extracts of AOiGHD mice, while expression of glucagon and somatostatin were increased. In order to determine if the reduction in insulin expression was due to a reduction in the number of [beta]-cells, formalin-fixed paraffin embedded sections of the pancreas from LF- and HF-fed control and AOiGHD mice were immunostained for insulin and mean islet size, islet number/pancreas, frequency of islet size and [beta]-cell mass/pancreas were assessed using image analysis software. The total number of islets/pancreas were significantly reduced in LF- and HF-fed AOiGHD mice, which could be attributed to a reduction in the number of small islets (1-30 cells/islet). However, this did not translate into a change in total [beta]-cell mass. Taken together, our results demonstrate that impaired glucose tolerance observed in diet-induced obese AOiGHD mice is not due to a deficit in [beta]-cell numbers. Therefore, in contrast to developmental GHD and GH resistant states, a reduction in circulating GH levels in adults may be more important in maintaining [beta]-cell function.[br][br](1) Liu JL, et al., Disruption of growth hormone receptor gene causes diminished pancreatic islet size and increased insulin sensitivity in mice. 2004. Am J Physiol Endocrinol Metab; 287(3):E405-13. (2) Wu Y, et al., Growth hormone receptor regulates [beta] cell hyperplasia and glucose-stimulated insulin secretion in obese mice. 2011. J Clin Invest; 121(6):2422-6. (3) Luque RM, et al., Metabolic impact of adult-onset, isolated, growth hormone deficiency (AOiGHD) due to destruction of pituitary somatotropes. 2011. PLoS One.; 6(1):e15767.[br][br]Sources of Research Support: Fundaci[oacute]n Alfonso Martin Escudero (J.C-C), Fundaci[oacute]n Caja Madrid (M.D.G.), , BFU2010-19300, RYC-2007-00186, CIBERObn (MICINN/FEDER), BIO-139, CTS-5051 (Junta de Andaluc[iacute]a) (R.L.M), Department of Veterans Affairs Merit Award and NIH R21AG031465 and R01DK088133 (R.D.K.).[br][br]Nothing to Disclose: JC-C, MDG, NP, RML, RDK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 447 296 2042 MON-107 PO13-01 Monday 1710 2012


1707 ENDO12L_MON-108 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) The Diabetes-Associated Gene [italic]Glis3[/italic] Regulates Beta Cell Differentiation In Utero Via [italic]Neurog3[/italic] Transactivation but Modulates Beta Cell Function in Adults Via Non-[italic]Neurog3-[/italic]Mediated Control of Insulin Production Yisheng Yang, Benny Chang, Lawrence Chan Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX [italic]Glis3[/italic], a member of the Kruppel-like family of transcription factors, is highly expressed in pancreatic beta cells. [italic]GLIS3[/italic] mutations were found to cause sporadic neonatal diabetes. Recent genome-wide association studies (GWAS) in adult populations identified [italic]GLIS3[/italic] also as a candidate gene for type 1 diabetes, and as a gene that is associated with type 2 diabetes. Moreover, the [italic]GLIS3[/italic] locus is linked to altered fasting glucose level in healthy children and adolescents. To gain insight into the functional roles of [italic]Glis3[/italic], we generated[italic] Glis3[/italic]null mice, which died neonatally with severe hyperglycemia and ketoacidosis. Using [italic]in vivo[/italic] and [italic]in vitro[/italic] analyses, we show that [italic]Glis3[/italic] is involved in the differentiation of endocrine progenitor cells through direct and indirect transcriptional control of [italic]Neurog3[/italic] expression, which is essential to islet formation. To determine if sustained [italic]Glis3[/italic] expression is required for the normal functioning of beta cells in adult animals, we generated conditional [italic]Glis3[/italic] knockout ([italic]Glis3[sup]fl/fl[/sup]/Pdx1Cre[sup]ERT+[/sup]) [/italic]mice and used tamoxifen to induce [italic]Glis3[/italic] deletion in adult mice. Tamoxifen-mediated beta cell-specific inactivation of [italic]Glis3[/italic] induces diabetes acutely by downregulating insulin expression, leading to hyperglycemia and subsequently enhanced beta cell apoptosis, an effect that appears to be largely independent of [italic]Glis3[apos]s[/italic] [italic]Neurog3[/italic] transactivating function. We conclude that [1] [italic]Glis3[/italic] controls fetal islet differentiation via [italic]Neurog3 [/italic]which perturbation causes neonatal diabetes, and [2] [italic]Glis3[/italic] expression is required for normal pancreatic beta cell function in adults, which impairment leads to defective insulin production and diabetes.[br][br]Sources of Research Support: This research was supported by the Diabetes and Endocrinology Research Center (P30DK079638), the Betty Rutherford Chair for Diabetes Research from St. Luke[apos]s Episcopal Hospital and the Charles and Barbara Close Foundation.[br][br]Nothing to Disclose: YY, BC, LC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 418 296 2043 MON-108 PO13-01 Monday 1711 2012


1708 ENDO12L_MON-109 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Caerulin Precursor Fragment (CPF-AM1): A Novel Non-Toxic Insulin-Releasing Peptide Isolated from the Skin Secretion of the Clawed Frog, [italic]Xenopus amieti[/italic] Opeolu O Ojo, John M Conlon, Peter R Flatt, Yasser HA Abdel-Wahab University of Ulster, Coleraine, UK; United Arab Emirates University, Al-Ain, United Arab Emirates We reported the isolation and structural characterization of CPF-AM1 and other peptides from the skin secretion of [italic]Xenopus amieti. [/italic]This study investigated the insulin-releasing activities of synthetic CPF-AM1 using clonal pancreatic cell line, BRIN-BD11 and Swiss TO mice with diet-induced insulin resistance. Acute insulin-release studies were performed in Krebs Ringer bicarbonate buffer supplemented with 5.6mM or 16.7mM glucose, purified synthetic peptide (0 [ndash] 3[micro]M) and known modulators of insulin secretion. Insulin-release was measured by radioimmunoassay. Membrane potential and intracellular calcium ([Ca[sup]2+[/sup]]i) were measured by flourometric assay using FLEXstation[sup]TM[/sup]. Degradation of CPF-AM1 by plasma enzymes was investigated using reversed-phase HPLC and MALDI-TOF spectrometer. At 5.6mM glucose, CPF-AM1 significantly stimulated insulin-release over concentration range of 30nM (1.4-fold, P[lt]0.05) to 3[micro]M (3.4-fold, P[lt]0.001) without beta-cell cytotoxicity. Without extracellular calcium, the stimulation was reduced by 25.3%. CPF-AM1(1[micro]M)-induced stimulatory effects were inhibited by co-incubation with 50[micro]M verapamil (57.4%, P[lt]0.001) and 300[micro]M diazoxide (51.8%, P[lt]0.001). Insulin-secretion increased by 3.3-fold with KCl (30mM, 16.7mM glucose) and CPF-AM1 (1[micro]M). At 5.6mM glucose and 1[micro]M CPF-AM1, co-incubation with 200[micro]M IBMX and 200[micro]M tolbutamide caused 1.2-fold and 1.1-fold increase in insulin-release respectively. The peptide induced membrane depolarization (2.6-fold) and increased [Ca[sup]2+[/sup]][sub]i[/sub] (2.0-fold) at 5.6mM glucose. [italic]In vivo[/italic], intraperitoneal administration of CPF-AM1 (75nmol/kg bw) with 18mmol/kg glucose significantly enhanced insulin-release (1.5-fold) and improved glucose tolerance by 28% (n=6, P[lt]0.05). CPF-AM1 is resistant to degradation by plasma proteolytic enzymes up to 8hr. The study showed that CPF-AM1 is a novel peptide with potential for development into a new antidiabetic drug.[br][br]Nothing to Disclose: OOO, JMC, PRF, YHAA-W 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 175 296 2044 MON-109 PO13-01 Monday 1712 2012


1709 ENDO12L_MON-110 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Plasma Taurine-Conjugated Bile Acid Concentrations Are Elevated in Type 2 Diabetes Marlene Wewalka, Florencia Halperin, Mary-Elizabeth Patti, Sander M Houten, Allison B Goldfine Joslin Diabetes Center, Harvard Medical School, Boston, MA; Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA; Academic Medical Center, Amsterdam, Netherlands Bile acids (BA) are essential for lipid metabolism and newly recognized as important signaling molecules in glucose and energy homeostasis, in part mediated by activation of peripheral G-protein coupled receptors and regulation of hepatic gluconeogenesis. Differences in BA profiles in patients with impaired glucose tolerance (IGT) and type 2 diabetes mellitus (T2D) remain incompletely understood. We performed 75g oral glucose tolerance tests (OGTT) in 99 persons with one or more risk factors for T2D, and classified groups based on glucose tolerance (normal glucose tolerance [NGT], n=62; IGT, n=25; and T2D n=12). Fasting BA (cholic acid [CA], chenodeoxycholic acid [CDCA], deoxycholic acid [DCA], ursodeoxycholic acid [UDCA], muricholic acid [MCA] and their respective taurine [T] and glycine [G] conjugates were measured by high-performance liquid chromatography tandem mass spectrometry, and quantified using deuterium-labeled internal standards. Group comparison was performed by ANOVA with post-hoc Tukey correction. All variables were logarithmically transformed if not normally distributed.[br]T2D subjects were older ([italic]P[/italic]=0.02), had higher systolic [italic](P[/italic]=0.007) and diastolic ([italic]P[/italic]=0.02) blood pressure, as well as higher fasting glucose and insulin concentrations, higher hemoglobin A1c, HOMA-IR and Disposition Index (all [italic]P[/italic][lt]0.001). IGT subjects were intermediate in these variables. Body mass index, total cholesterol and triglyceride levels were similar between groups.[br]Plasma total BA and total glycine-conjugated BA were higher in T2D, but these differences did not reach statistical significance. Total taurine-conjugated BA (total T-BA) ([italic]P[lt][/italic]0.01), and its subsets of T-UDCA ([italic]P[/italic]=0.03) and T-DCDA/T-DCA ([italic]P[/italic]=0.03) differed between groups, with higher concentrations in T2D than NGT, and intermediate in IGT. T-CA trended higher ([italic]P[lt][/italic]0.06). Simple regression revealed a positive correlation between total T-BA and fasting glucose (R=0.37, [italic]P[/italic][lt]0.0001), total T-BA and 2-hour glucose after OGTT (R=0.31, [italic]P[/italic]=0.002) and total T-BA and HOMA-IR (R=0.26, [italic]P[/italic]=0.01), as well as a negative relationship between total T-BA and oral Disposition Index (R=-0.36, [italic]P[/italic]=0.001).[br]Fasting taurine-conjugated BA concentrations are higher in T2D and intermediate in IGT, compared to NGT. Taurine-conjugated BA correlate with fasting and post-load glycemia. Further study is needed to elucidate whether this pattern of taurine-BA conjugation may be targeted for novel therapeutic approaches to treat T2D.[br][br]Sources of Research Support: NIH DK062948 (M.-E.P), DK060837 (M.-E.P and A.B.G), Juvenile Diabetes Research Foundation (M.-E.P and A.B.G), K23-DK02795 and American Diabetes Association Career Development Award 06-CD-07 (A.B.G),Marietta Blau Grant ICM-2010-02797 (M.W).[br][br]Nothing to Disclose: MW, FH, M-EP, SMH, ABG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1739 296 2045 MON-110 PO13-01 Monday 1713 2012


1710 ENDO12L_MON-111 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Apelin Regulates Pancreatic [beta]-Cell Homeostasis Song Han, Ella W Englander, George H Greeley, Jr University of Texas Medical Branch, Galveston, TX Apelin is a small regulatory peptide cloned from bovine stomach extracts based upon its ability to activate the APJ receptor. Apelin exerts several metabolic effects including regulation of lipolysis and maintenance of insulin sensitivity. Apelin and APJ are expressed in pancreatic [beta]-cells and apelin has been shown to influence insulin secretion. Whether apelin affects pancreatic [beta]-cell homeostasis is not known. The aim of this study, therefore, was to define the influence of apelin on pancreatic [beta]-cell homeostasis. [bold]Methods:[/bold] Adult male mice were treated with either vehicle or synthetic pyro-apelin-13 (25 ug/injection, 2x/day for 4 days, IP). Pancreases were harvested, cleared of extraneous tissue and weighed. Pancreases were fixed and sectioned. For determination of islet size and mass, every islet in three pancreatic sections for each mouse were immunostained for insulin, N= 5-7 mice/group. Individual islet areas and total pancreatic section areas were measured. [bold]Results:[/bold] Pancreatic weights (mean [plusmn] SEM) were unaffected by apelin treatment (vehicle: 299[plusmn]15 vs. apelin-treated: 295[plusmn]9 mg). Apelin treatment increased the fractional islet area (% of pancreas, vehicle: 1.1[plusmn]0.15 vs. apelin-treated: 1.9[plusmn]0.40), [beta]-cell mass (BCM) (vehicle: 3.0[plusmn]0.4 vs. apelin-treated: 5.5[plusmn]1.2 mg) and islet density (number of islets/mm[sup]2 [/sup]pancreas area) (vehicle: 2.9[plusmn]0.4 vs. apelin-treated: 3.8[plusmn]0.40) significantly (P[lt]0.05). Mean islet size is increased but not significantly in apelin-treated mice (vehicle: 3602[plusmn]531 vs. apelin-treated: 4441[plusmn]443 um[sup]2[/sup]). [bold]Conclusions and Discussion:[/bold] Our findings imply that apelin exposure increases fractional islet area and BCM by increasing islet density and islet size. Apelin may increase BCM by stimulating [beta]-cell proliferation, reducing apoptosis or by both mechanisms. These results indicate that apelin is a potential islet and [beta]-cell growth factor that can be exploited to treat pancreatic endocrine insufficiency.[br][br]Nothing to Disclose: SH, EWE, GHG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 209 296 2046 MON-111 PO13-01 Monday 1714 2012


1711 ENDO12L_MON-112 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) The Mitogen-Activated Protein Kinase p38 Acutely Regulates the Translation of iNOS mRNA in Islet [beta] Cells in Response to Cytokine Stress Yurika Nishiki, Andrew T Templin, Bernhard Maier, Raghavendra G Mirmira Indiana University School of Medicine, Indianapolis, IN In types 1 and 2 diabetes, production of proinflammatory cytokines acutely activate inducible nitric oxide synthase (iNOS) to generate nitric oxide, which is thought to cause islet beta cell dysfunction and impairments in insulin secretion. Although the regulation of iNOS mRNA transcription has been studied extensively, the post-transcriptional mechanisms remain poorly defined. It is known that the mitogen-activated kinases (MAPKs) such as p38, JNK, and ERK are responsive to cytokine stress, but their roles (if any) in translation are unknown. Additionally, our prior studies (1) showed that in beta cells iNOS mRNA translation is dependent upon the action of deoxyhypusine synthase, an enzyme that is necessary for the activity of the translational factor eIF5A. To determine whether MAPKs are linked to translational regulation via eIF5A, we studied iNOS mRNA translation in INS-1 (832/13) beta cells and rat islets. When beta cells were challenged by cytokine stress, we observed rapid phosphorylation and translocation of NFkB p65 into the nucleus, followed by a 3-4 order of magnitude induction of iNOS mRNA. These events are not inhibited by treatment of cells with any MAPK inhibitors. However, a striking 60% inhibition of iNOS protein was observed in the presence of the p38 MAPK inhibitor PD169316. Whereas cytokines induced a 2-fold increase in eIF5A hypusination, pretreatment with PD169316 abrogated this effect, coincident with the block in iNOS protein production. To assess directly the possible translational regulation of iNOS mRNA, we performed polyribosomal profiling experiments using sucrose gradient sedimentation of ribosome associated RNA from INS-1 cells. Under cytokine stress, iNOS mRNA was associated with polyribosomes indicating active iNOS translation. Upon treatment with PD169316 we observed a shift of the iNOS mRNA from polyribosomal fractions into the monoribosomal fractions, indicating a reduction in translation initiation and/or elongation. We conclude that in response to cytokine stress, p38 MAPK acutely promotes the translation of iNOS mRNA, possibly via the regulation of eIF5A hypusination and activity.[br][br](1) Maier B et al., J Clin Invest. 2010;120(6):2156[ndash]2170.[br][br]Sources of Research Support: NIH, R01, DK60581.[br][br]Nothing to Disclose: YN, ATT, BM, RGM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2192 296 2047 MON-112 PO13-01 Monday 1715 2012


1712 ENDO12L_MON-113 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Proinflammatory Cytokines Acutely Impair Islet Beta Cell Oxygen Consumption Rate by Inhibition of Mitochondrial Aconitase Activity Adeola Adewola, Bernhard Maier, Raghavendra Mirmira Indiana University School of Medicine, Indianapolis, IN The release of insulin by the islet beta cell is dependent on the generation of ATP via glycolytic and tricarboxylic acid (TCA) cycle pathways. In the pathogenesis of both type 1 and type 2 diabetes, impaired insulin release by the beta cell is thought to be caused by proinflammatory cytokines, which interfere with beta cell production of ATP through mechanisms that are not well defined. To clarify potential mechanisms of impaired ATP production in the presence of cytokines, we used real-time metabolic flux analysis to monitor changes that acutely follow exposure of beta cells to proinflammatory cytokines (TNF-alpha, IFN-gamma, IL-1beta). This type of analysis involves simultaneous measurements of extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) to assess the state of glycolysis and mitochondrial respiration, respectively. Rat INS-1 (832/13) beta cells were pre-incubated with cytokines for 4 hours and then subjected to metabolic flux analysis. Whereas prolonged (more than 24 h) exposure to cytokines resulted in very severe impairments in both ECAR (glycolysis) and OCR (mitochondrial respiration), short-term (4 h) cytokine challenge had no effect on glucose-stimulated ECAR but resulted in an inhibition in glucose-stimulated OCR. Published studies suggest that nitric oxide or peroxynitrite can impair enzymes of the TCA cycle by S-nitrosylation or nitration, respectively. Consistent with the possibility that NO was the offending molecule, inhibition of inducible nitric oxide synthase with L-NMMA or blocking its mRNA translation by inhibiting deoxyhypusine synthase (with GC7) restored the impairment in glucose-stimulated OCR in response to cytokines. Supplementation with 1 mM isocitrate (a product of actonitase activity) also completely restored OCR in the presence of cytokines, suggesting that mitochondrial aconitase is a direct or indirect target of NO. We conclude that proinflammatory cytokines acutely disrupt beta cell mitochondrial respiration via production of NO, which subsequently causes inhibition of mitochondrial aconitase activity.[br][br]Sources of Research Support: NIH grant R01 DK083583 and JDRF.[br][br]Nothing to Disclose: AA, BM, RM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2266 296 2048 MON-113 PO13-01 Monday 1716 2012


1713 ENDO12L_MON-114 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Cell-Autonomous Function of Bmal1 Is Required To Preserve [beta]-Cell Function Jeongkyung Lee, Mi-Sun Kim, Rongying Li, Victoria Y Liu, Zhe Fang, David L Nelson, Loning Fu, David D Moore, Ke Ma, Vijay K Yechoor Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; The Methodist Hospital Research Institute, Houston, TX Circadian misalignment, with shift work, leads to diabetes, obesity and metabolic syndrome. We have shown that genetic disruption of the circadian clock by global deletion of Bmal1, a non-redundant core clock gene in mice, leads to diabetes secondary to [beta]-cell failure due to defects in [beta]-cell stimulus-secretion coupling and glucose-stimulated insulin secretion (GSIS). To investigate if this impairment is due to a loss of cell-autonomous function of Bmal1in [beta]-cells, we generated mice with a disrupted [beta]-cell clock ([beta]-Bmal1-/-) by crossing floxed Bmal1 (Bmal1 F/F) and Rat insulin promoter-cre (Rip-cre) transgenic mice.[br][beta]-Bmal1-/- mice display absence of nuclear Bmal1 protein only in [beta]-cells, but not in SCN or other tissues and have an intact central clock, demonstrated by normal wheel running activity. [beta]-Bmal1-/- mice develop glucose intolerance with blunted GSIS, demonstrating that the cell-autonomous function of Bmal1 is required for normal [beta]-cell function. The blunted GSIS is secondary to an upregulation of Ucp2, increased mitochondrial uncoupling and consequent impairment of glucose-induced mitochondrial hyperpolarization and ATP production. Inhibition of Ucp2 in isolated islets rescues the impaired GSIS. These changes are due, in part, to increased accumulation of reactive oxygen species resulting from decreased expression of the anti-oxidant genes in [beta]-Bmal1-/- islets. Chromatin immunoprecipitation reveals Bmal1 occupancy in the promoters of these genes, suggesting direct regulation.[br]We then tested if [beta]-cell clock is required to preserve [beta]-cell function with circadian misalignment by exposing [beta]-Bmal1-/- and control Bmal1 F/F mice to either normal 12 hr light/dark (LD [ndash] lights-6AM to 6PM) cycle or alternating phase advance and delay of LD cycle (3days with normal LD alternating with 4 days with a 6 hr shift; lights-12:01AM to 12:01PM). Interestingly, control mice display blunting of first phase insulin secretion, suggesting [beta]-cell dysfunction, after 8 wks of circadian misalignment, while there was significant worsening of glucose tolerance and abrogation of first phase insulin secretion in [beta]-Bmal1-/- mice.[br]Taken together, the cell-autonomous function of the [beta]-cell clock is required for preserving [beta]-cell function under normal and circadian disrupted conditions. Understanding the underlying mechanisms would lead to insights, of potential therapeutic impact, into the pathophysiology of [beta]-cell failure and diabetes.[br][br]Nothing to Disclose: JL, M-SK, RL, VYL, ZF, DLN, LF, DDM, KM, VKY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1841 296 2049 MON-114 PO13-01 Monday 1717 2012


1714 ENDO12L_MON-115 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Morphometric Analysis of Langerhans Islets in Mice Deficient in the [italic]Slc12a2[/italic] Gene Elizandra de Sousa da Silva, Saeed Alshahrani, Mauricio Di Fulvio Wright State University, Dayton, OH Mice deficient in different genes, commonly called knockout mice (KO), have been widely used to determine the functional impact that genes may have in the organism and to explore their function at the organismal level. Mice with homozygous deficiency in the [italic]Slc12a2[/italic] gene exhibit unique endocrine and behavioral characteristics such as relatively flat glucose tolerance curves, borderline hypoglycemia, hyperphagia, hypoinsulinemia, food aggression and low physical development. The [italic]Slc12a2[/italic] gene encodes for a membrane protein, known as the Na[sup]+[/sup]K[sup]+[/sup]2Cl[sup][ndash][/sup] co-transporter 1. [italic]Slc12a2[/italic] plays a key role in cell volume regulation and in keeping the intracellular chloride concentration above electrochemical equilibrium in almost all mammalian cells studied so far. In spite of the very well known function of this gene product, virtually nothing is known regarding the role of [italic]Slc12a2[/italic] in endocrine function. To gain insights into the potential mechanisms involved in altered glucose homeostasis observed in these transgenic mice, the number of [beta] cells per islet and the islet density were morphometrically determined in pancreas[apos] slides of [italic]Slc12a2[/italic][sup]KO[/sup] and [italic]Slc12a2[/italic][sup]WT[/sup] mice immunostained against insulin. Our results show that [italic]Slc12a2[/italic][sup]KO[/sup] islets have significantly more [beta] cells than [italic]Slc12a2[/italic][sup]WT[/sup] mice. Further, our analysis revealed higher islet density and lower pancreas mass in [italic]Slc12a2[/italic][sup]KO[/sup] mice when compared to [italic]Slc12a2[/italic][sup]WT[/sup] littermates. These anatomical changes correlated with increased insulin response post-glucose load. Therefore, our results suggest that mice deficient in the [italic]Slc12a2[/italic] gene have an increased insulin secretory capacity explaining their unusual glucose homeostasis profile and behavior.[br][br]Sources of Research Support: This work was supported by funds awarded to MDiF by the American. Diabetes Association (Grant 1-10-JF-14).[br][br]Nothing to Disclose: EdSdS, SA, MDF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1981 296 2050 MON-115 PO13-01 Monday 1718 2012


1715 ENDO12L_MON-116 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Transcriptional Activation of Reg2 and Reg3[beta] Genes by Glucocorticoids in Rodent Pancreatic Acinar and Insulinoma Cells Chen Luo, Bing Li, Min Wang, Jun-Li Liu McGill University Health Centre, Montr[eacute]al, Canada; China Pharmaceutical University, Nanjing, China Reg family genes are expressed in pancreatic acinar and ductal cells and involved in pancreatitis and islet [beta]-cell growth. Among the 7 encoded proteins, Reg1 and Reg3[delta] (INGAP) stimulate the proliferation or regeneration of the islet cells(1). Conversely, their level of expression is activated by nutrients, cytokines and other hormones but inhibited by glucocorticoids(2). We have reported that overexpressed Reg2 and Reg3[beta] protect mouse [beta]-cells against streptozotocin-induced damage(3, 4); however, the mechanism of their gene regulation has been elusive. (1) In order to explore transcriptional regulation, we transfected pGL3-Basic, luciferase reporter genes driven by 1.8-kb promoters of mouse origin into rat acinar derived AR42J cells. The promoter expression of both genes was significantly [italic]activated[/italic] for a maximum 25 fold by dexamethasone (DEX) in a dose-dependent manner; similar response was not seen using the promoters of Reg3[alpha] and Reg3[delta]. Promoter deletions revealed putative glucocorticoids responsive elements within -270/-675 bp of Reg2 and -288/-1051 bp of Reg3[beta] promoters respectively. To validate its biological relevance, the levels of endogenous Reg3[beta] mRNA and protein in AR42J cells were both increased significantly by DEX; Reg2 was not expressed in the rat cells; in contrast, DEX caused a drastic [italic]decrease[/italic] in the level of Reg1. Finally, unlike Reg1 gene being activated by nicotinamide(5, 6), DEX-induced Reg2 and Reg3[beta] promoter activation was dose-dependently [italic]inhibited[/italic] by this inhibitor of poly(ADP-ribose) polymerase (Parp). Our results for the first time indicated a significant and direct [italic]stimulation[/italic] of Reg2 and Reg3[beta] genes by glucocorticoids, and the effect was [italic]abolished[/italic] by Parp inhibitor. The secretion of glucocorticoids, Reg1 and Reg3[beta] are all activated in response to inflammation (such as in pancreatitis), prominent stimulation of specific Reg genes by glucocorticoids may constitute a synergistic anti-inflammatory mechanism. (2) In mouse insulinoma MIN6 cells, DEX stimulated the transcription of Reg3[beta] promoter in a dose-dependent manner, but not that of Reg2, 3[alpha] or 3[delta], indicating strict cell- and promoter-specific effects. While IL-6 and nicotinamide alone had no direct effect, coculture of IL-6 and DEX produced a drastic synergism on Reg3[beta] gene transcription, which was abolished by the addition of nicotinamide. The result indicates that endogenous Reg3[beta] could be activated in MIN6 cells under inflammatory conditions to protect the [beta]-cells.[br][br]1. Liu JL, et al. Endocr Metab Immune Disord Drug Targets 2007; 8:1. 2. Zenilman ME, et al. Ann Surg 1997; 225:327. 3. Liu L, et al. Growth Factors 2010; 28:370. 4. Xiong X, et al. Am J Physiol Endocrinol Metab 2011; 300:E669. 5. Terazono K, et al. J Biol Chem 1988; 263:2111. 6. Akiyama T, et al. Proc Natl Acad Sci USA 2001; 98:48.[br][br]Sources of Research Support: Canadian Institutes of Health Research MOP-84389; Canadian Diabetes Association OG-3-11-3469-JL; Chercheurs-boursier of Fonds de la recherche en sante Quebec;China Scholarship Council 2010706006.[br][br]Nothing to Disclose: CL, BL, MW, J-LL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1063 296 2051 MON-116 PO13-01 Monday 1719 2012


1716 ENDO12L_MON-117 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Lineage Tracing of Beta Cells during Dedifferentiation to Duct Epithelium: Involvement of Insulin+ Progenitor Cells Christine A Beamish, Brenda J Strutt, David J Hill Western University Canada, London, Canada; Lawson Research Institute, London, Canada; Western University Canada, London, Cape Verde Regeneration of pancreatic beta cells as a strategy for the reversal of diabetes is dependent on an understanding of the origins of beta cells and their plasticity. Developmentally, islets arise from cells in the pancreatic duct epithelium. Dedifferentiation of [beta]-cells [italic]in vitro[/italic] back to a more proliferative duct phenotype could be induced and expanded before redifferentiation to an insulin-producing population. We hypothesize that mouse islet dedifferentiation to duct epithelium will involve participation of the [beta]-cell.[br]We have developed [italic]in vitro[/italic] protocols for the culture of neonatal mouse islets on collagen in the presence of EGF and cholera toxin to an epithelial phenotype. Genetic analysis by RT-PCR amplification and immunofluorescent confocal microscopy were used to determine differentiation states. We have employed a transgenic mouse strain, RIPCre;Z/AP, that allows tagging of [beta]-cells with a human placental alkaline phosphatase (HPAP) reporter protein to track the fate of [beta]-cells during culture and dedifferentiation.[br]Islets plated for dedifferentiation lost all morphological, genetic, and protein markers of mature function, including insulin and Pdx1 expression. Proliferation increased significantly. Markers of duct cells (Carbonic anhydrase, Cytokeratin19 (CK19)) and ductal precursors (Hnf6, Hnf1[beta], Sox9) were expressed. Expression of the beta cell reporter (HPAP) was decreased drastically after four weeks in culture, and these cells were not proliferative. The few remaining HPAP[sup]+[/sup] (former [beta]) cells however, gained CK19 expression. Recently, rare insulin[sup]+[/sup] progenitor cells have been reported, identified by the absence of a glucose transporter (Glut2). Glut2[sup]-[/sup]Ins[sup]+[/sup] cells from pancreata of RIPCre;Z/AP[sup]+/+[/sup] mice were isolated using flow cytometry, and cultured for dedifferentiation. After four weeks, a large number of the progenitor cells had differentiated into CK19[sup]+[/sup] epithelium while maintaining expression of HPAP; Glut2[sup]+[/sup]Ins[sup]+[/sup] [beta]-cells did not exhibit the same pattern.[br]These data indicate that CK19[sup]+[/sup] duct epithelial cells arose from isolated neonatal mouse islets in our [italic]in vitro[/italic] model. It remains to be elucidated whether the HPAP[sup]+[/sup]CK19[sup]+[/sup] cells seen are derived from dedifferentiated mature [beta]-cells, or alternately from the activation of inherent insulin[sup]+[/sup] progenitor cells, as well as the relative contributions of each population to endocrine pancreas plasticity.[br][br]Nothing to Disclose: CAB, BJS, DJH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2326 296 2052 MON-117 PO13-01 Monday 1720 2012


1717 ENDO12L_MON-118 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Human Amylin Transgenic Mice Show Elevated Core Body Temperature and Increased Energy Expenditure Jacqueline F Aitken, Christopher S Walker, Gordana Prijic, Sarah Johnson, Garth JS Cooper University of Auckland, Auckland, New Zealand; University of Auckland, Auckland, New Zealand; University of Manchester, Manchester, UK [bold]Introduction.[/bold][br]Human amylin (hA), also called Islet Amyloid Polypeptide (IAPP) is a 37 amino acid protein that is co-secreted with insulin from the pancreatic islet [beta]-cells upon stimulation with glucose (1,2). Aggregation of hA/IAPP into small soluble [beta]-sheet-containing oligomers is linked to islet [beta]-cell degeneration and the pathogenesis of type 2 diabetes (3).[br]We have previously reported the characterisation of an hA/IAPP transgenic mouse line, which overexpresses human amylin in the pancreas and spontaneously develops diabetes (4).[br]Amylin/IAPP reduces food intake after peripheral or central administration. It functions as both a short-term satiation signal and also has long-term anorectic and weight reducing actions. Studies have reported that amylin/IAPP may increase energy expenditure and increase body temperature, although the influence of amylin/IAPP on body temperature is still questionable.[br][bold]Purpose.[/bold][br]This study sought to investigate whether there was a relationship between expression of the human amylin/IAPP protein, and metabolic and physiological parameters such as core body temperature, energy expenditure, weight, blood glucose and selected hormone levels in hA/IAPP transgenic mice.[br][bold]Methods.[/bold][br]Blood Glucose, body weight, core body temperature were measured weekly and indirect calorimetry was performed in approximately 100 day old hA/IAPP transgenic and control littermates using the Comprehensive Laboratory Animal Monitoring System (CLAMS). Food intake, activity, O[sub]2[/sub] consumption (VO[sub]2[/sub]), CO[sub]2[/sub] production (VCO[sub]2[/sub]) were measured and respiratory quotients and energy expenditures were calculated. Serum hormone levels, including insulin, amylin, leptin and adiponectin, were measured in control and transgenic mice.[br][bold]Results and Conclusions.[/bold][br]Weight, blood glucose levels, core body temperatures, food intake, activity, respiratory quotient (RQ) and energy expenditure results are presented along with serum hormone and liver glycogen levels in the hemizygous hA/IAPP transgenic mice, compared with their non-transgenic littermates.[br]Hemizygous hA/IAPP transgenic mice had significantly higher core body temperatures and energy expenditure compared to their non-transgenic littermates, independent of the diabetic state of the transgenic animal.[br]Adiponectin levels in serum were significantly lower and the liver contained increased glycogen content in the hemizygous hA/IAPP transgenic mice compared to their control littermates.[br][br](1)Cooper,GJS et al.,Proc.Natl.Acad.Sci.(USA) 1987; 84:8628. (2)Westermark,P et al.,Proc.Natl.Acad.Sci.(USA) 1987; 84:3881. (3)Cooper,GJS et al.,Diabetologia 2010; 53:1011. (4)Aitken,JF,Loomes,KM et al.,Diabetes 2010; 59:161.[br][br]Sources of Research Support: Endocore Research Trust; Maurice and Phyllis Paykel Trust; New Zealand Ministry of Science and Innovation; New Zealand Lotteries Commission.[br][br]Nothing to Disclose: JFA, CSW, GP, SJ, GJSC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 527 296 2053 MON-118 PO13-01 Monday 1721 2012


1718 ENDO12L_MON-119 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Examining the Role of Connective Tissue Growth Factor (CTGF) in beta-Cell Replication and Pancreatic Islet Transplantation Uma Gunasekaran, Maureen A Gannon Vanderbilt University Medical Center, Nashville, TN CTGF (connective tissue growth factor) is a secreted protein that modulates TGF-beta, BMP, and Wnt signaling. In the pancreas, CTGF is expressed in embryonic beta cells, ductal epithelium, and pancreatic vasculature but is silenced in beta cells, after birth. In other cell types, CTGF plays a role in cell migration and cell proliferation, as well as vasculogenesis. We previously showed that CTGF null mutant mouse embryos have increased alpha cell mass and a concomitant reduction in beta cell mass due to decreased beta cell proliferation compared to wild-type embryos. We hypothesize that CTGF over-expression in adult mice will increase proliferation and subsequently beta cell mass. To test this hypothesis, we generated mice in which CTGF over-expression in beta cells is doxycycline-dependent. We have already shown that CTGF over-expression in embryos causes an increase in endocrine mass and beta cell proliferation. CTGF expression was induced with doxycycline in 6-7 week old mice for various time periods (1-10 weeks). Glucose tolerance tests revealed that CTGF over-expression has no effect on glucose homeostasis at any time point examined compared to control littermates. Currently, samples of pancreata are being examined histologically for the effect of CTGF over-expression on beta cell proliferation and mass and islet vascularization. In addition, CTGF has been induced in the setting of islet transplantation to determine whether this factor would be beneficial for beta cell survival and islet revascularization.[br][br]Sources of Research Support: JDRF Grant 1-2011-592; NRSA 1F32DK092001-01.[br][br]Nothing to Disclose: UG, MAG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1616 296 2054 MON-119 PO13-01 Monday 1722 2012


1719 ENDO12L_MON-120 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Regulation of Insulin Secretion by Vesicular Nucleotide Transporter Mediated ATP Signaling Jessica Catherine Geisler, Kathryn Lynn Corbin, Craig Nunemaker, Chien Li University of Virginia, Charlottesville, VA; University of Virginia, Charlottesville, VA Type 2 diabetes (T2D) is a multifactorial disorder characterized by dysregulated insulin secretion and insulin resistance in peripheral tissues. T2D may be caused by a number of lifestyle factors, yet obesity is the primary risk factor in individuals with a genetic predisposition. Consuming a high fat diet leads to excessive energy intake and consequent obesity. Currently the mechanisms underlying [beta] cells dysfunction associated with excessive nutrient intake and obesity remain elusive. We have found elevated levels of vesicular nucleotide transporter (VNUT) in MIN6 cells, a mouse [beta] cell line, treated with high fat media. Likewise, we also discovered increased VNUT expression in pancreatic islets isolated from mice fed a high fat diet. VNUT is a recently identified vesicular transporter in adrenal chromaffin cells and is responsible for transporting ATP into secretory vesicles for storage and release. It has been shown that ATP is present in insulin secretory vesicles and that ATP can regulate insulin release via purinergic signaling; however, the mechanism of ATP accumulation into insulin secretory vesicles remains unknown. To probe the functional role of elevated VNUT expression in pancreatic [beta] cells, we generated a VNUT overexpressing lentivirus. Overexpressing VNUT in [beta] cells resulted in enhanced ATP release, which was measured by luciferase assay. Furthermore, overexpression of VNUT in isolated mouse islets resulted in elevated basal insulin levels, but reduced sensitivity to glucose-stimulated insulin secretion (GSIS), as determined by an insulin enzyme-linked immunosorbent assay. Moreover, pretreating islets with suramin, a purinergic receptor antagonist, completely reversed enhanced basal insulin secretion induced by VNUT overexpression. On the other hand, knocking down (KD) VNUT expression with a VNUT-specific short hairpin RNA (shRNA) lentivirus suppressed ATP release from [beta] cells. Intriguingly, we saw a dramatic suppression of both basal and GSIS in VNUT shRNA lentivirus infected islets, yet treating VNUT KD islets with ATP significantly recovered insulin secretion. This suggested that reduced insulin secretion observed in VNUT KD islets is due, at least in part, to decreased extracellular ATP signaling. In summary, our study reveals that VNUT is expressed on insulin secretory vesicles in pancreatic [beta] cells and plays an important and novel role in ATP release which consequently modulates insulin secretion through purinergic signaling.[br][br]Nothing to Disclose: JCG, KLC, CN, CL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 996 296 2055 MON-120 PO13-01 Monday 1723 2012


1720 ENDO12L_MON-121 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Transgenic Activin-A Regulates Adult Pancreatic Islet Gene Expression and Protects Against Hyperglycemia Ezra M Wiater, Wylie W Vale The Salk Institute for Biological Studies, San Diego, CA Activin-A is a member of the activin sub-family within the TGF-beta superfamily of growth factors, which also includes the BMP and TGF-beta families. Activins are widely expressed hormones, with important roles in reproduction, through their actions on FSH, and numerous other actions in development and in adult homeostasis. Mice with loss or over-expression of activin-A, activin receptors, or activin signal transducing Smad proteins exhibit developmental defects in the formation of pancreatic islets and disruptions in adult beta-cell mass, consistent with activin-A effects in pancreas and islet development. However, the actions of activin-A in adult pancreas are less clear.[br]In order to investigate activin-A effects in adult pancreas, we generated beta-cell specific doxycycline-inducible transgenic mice to allow over-expression of activin-A or the soluble activin binding protein follistatin in adult pancreatic beta-cells. We confirmed that doxycycline treatment resulted in increased expression of the human activin-A transgene in mouse islets. Induced transgenic expression of activin-A modulated the expression of endogenous pancreatic target genes in isolated primary islets and in intact pancreas, confirming that transgenic activin-A was bioactive. Effects of induced activin-A transgene expression in transgenic islets, such as the induction of the activin target gene Pax4, were similar to effects of treating primary islets with purified activin-A.[br]To determine the effects of activin-A on adult pancreatic beta-cell mass, we relied on a model of chemically-induced beta-cell damage using the beta-cell specific toxin streptozotocin. Activin-A over-expression following streptozotocin challenge protected adult mice from hyperglycemia, compared to wild-type mice or transgenic mice expressing the activin antagonist follistatin. In intact mice, induction of activin-A increased the expression of Pax4 and other pancreatic genes involved in specification and proliferation of beta -cells, as well as other islet specific genes.[br]Together, these data are consistent with activin-A having a role in the regulation of adult pancreatic islet cell types and improve beta-cell function by targeting multiple islet cell types and facilitating promotion of beta-cell proliferation or protection of beta-cell mass. These activin-A effects in adult pancreas could be exploited in the development of therapies to remedy beta-cell deficit, including type 1 and type 2 diabetes.[br][br]Sources of Research Support: The Clayton Medical Research Foundation, Inc. and a Sanofi Discovery Grant. WV was a senior CMRF, Inc. Investigator.[br][br]Disclosures: WWV: Wylie Vale was a co-founder, consultant, equity holder, membero of the Board of Directors and Scientific Advisory Board., Acceleron Pharma, Neurocrine Biosciences. Nothing to Disclose: EMW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2062 296 2056 MON-121 PO13-01 Monday 1724 2012


1721 ENDO12L_MON-122 POSTER SESSION: Islet Biology [amp] Insulin Secretion (1:30 PM-3:30 PM) Activin B Regulates Islet Composition and Function but Not Whole-Body Glucose Homeostasis or Insulin Sensitivity Lara Bonomi, Melissa L Brown, Nathan Ungerleider, Meghan E Muse, Martin M Matzuk, Alan L Schneyer UMass-Amherst, Springfield, MA; Baylor College of Medicine, Houston, TX Activins A and B are members of the TGF[beta] superfamily that have at least partially distinct activities [italic]in vivo[/italic]. The roles of activin A and B in regulating pancreatic islet function and whole body glucose homeostasis is receiving increased attention, with activin A having been demonstrated to enhance glucose stimulated insulin secretion (GSIS) in rat and human islets. Both activins utilize ACVR2B and ALK4 receptors for signaling, but activin B can also use ALK7. ALK7 deficient mice have elevated insulin concentrations suggesting that activin B opposes previously reported positive effects of activin A on glucose-stimulated insulin release. We hypothesized that activin B null mice (BBKO) would have enhanced glucose tolerance and their islets should secrete more insulin. We tested this hypothesis by analyzing glucose homeostasis, insulin production, and islet function and composition in activin B null mice. There were no differences in fasting glucose or insulin serum concentrations, nor in glucose tolerance or insulin sensitivity comparing weight-matched young or older BBKO and wildtype males. Similarly, there were no significant differences in insulin secretion comparing islets from wildtype or BBKO males at either age. However, BBKO islets were more sensitive to activin A, myostatin (MSTN), and follistatin (FST) treatments so that activin A and FST inhibited and MSTN enhanced glucose stimulated insulin secretion. While islet size was not different between the genotypes, [beta]-cell mass and the proportion of [alpha]-cells/islet were significantly reduced in BBKO islets. [italic]Pdx1[/italic] expression was significantly increased in BBKO islets as was [italic]MafB[/italic] while [italic]MafA[/italic] was reduced. Taken together, these results indicate that activin B does not antagonize activin A to influence whole body glucose homeostasis but does alter islet response to activin, MSTN and FST. In addition, activin B appears to modulate [alpha]-cell proliferation, [alpha]- to [beta]-cell transdifferentiation, and/or [beta]-cell survival since BBKO islets had an increased proportion of [beta]-cells/islet but overall reduction in [beta]-cell mass. Our results also show for the first time that MSTN can stimulate GSIS. However, this was only observed in BBKO islets, perhaps due to their increased sensitivity to ligands signaling through the activin receptor pathway resulting from the absence of activin B.[br][br]Sources of Research Support: National Institutes of Health, NIDDK R01 grant R01DK075058.[br][br]Nothing to Disclose: LB, MLB, NU, MEM, MMM, ALS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1603 296 2057 MON-122 PO13-01 Monday 1725 2012


1722 ENDO12L_MON-123 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Body Weight and Indices of Adiposity Are Similarly Affected by the Two Common Types of Fructose-Containing Sugars, Sucrose and High-Fructose Corn Syrup, When Consumed as Part of a Normal Diet Zhiping Yu, Joshua Lowndes, Von Nguyen, Stephanie Sinnett, Shannon Weston, James M Rippe Rippe Lifestyle Institute, Celebration, FL It has been postulated that excess fructose consumption may be obesigenic, particularly when consumed with glucose. Sucrose and high fructose corn syrup (HFCS) are the primary sources of fructose in the American diet. Despite acute studies showing them to have equivalent metabolic effects, few data exist on the longer-term metabolic effects of these two sugars when consumed at levels typical of the general population.[br]As part of their usual diet, sixty-four individuals (42.2 [plusmn] 11.7 years) consumed low-fat milk sweetened with either HFCS or sucrose in an amount that the added sugar consisted of 8%, 18% or 30% of total caloric intake for ten weeks. These levels represent the 25th, 50th and 95th percentile levels of fructose consumption in the American diet.[br]There were no significant increases in body mass at any level of added sugar consumption (p[gt]0.05). BMI, body fat percentage, waist circumference and fat mass also were unchanged in any group (p[gt]0.05). None of these measures were influenced by the type or amount of added sugar consumed.[br]HFCS and sucrose have comparable effects and do not cause changes in body weight or indices of adiposity when consumed at up to the 95th percentile population consumption level.[br][br]Sources of Research Support: Corn Refiners Association.[br][br]Nothing to Disclose: ZY, JL, VN, SS, SW, JMR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 131 297 2058 MON-123 PO28-01 Monday 1726 2012


1723 ENDO12L_MON-124 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Central Adiposity and Metabolic Profile Are Similarly Affected by the Two Common Types of Fructose-Containing Sugars, Sucrose and High-Fructose Corn Syrup, When Consumed as Part of a Normal Diet Joshua Lowndes, Zhiping Yu, Stephanie Sinnett, Von Nguyen, Shannon Weston, James Rippe Rippe Lifestyle Institute, Celebration, FL It has been postulated that excess fructose consumption may be more problematic than similar amounts of other sugars with respect to promoting obesity, particularly in the abdominal region, and result in a greater worsening in metabolic profile. Sucrose and high fructose corn syrup (HFCS) are the primary sources of fructose in the American diet, and despite acute studies showing them to have equivalent metabolic effects, few data exist on the longer-term metabolic effects of these two sugars when consumed at levels typical of the general population.[br]As part of their usual diet, sixty-four individuals (42.2 [plusmn] 11.7 years) consumed low-fat milk sweetened with either HFCS or sucrose in an amount that the added sugar consisted of 8%, 18% or 30% of total caloric intake for ten weeks. These levels represent the 25th, 50th and 95th percentile population consumption levels of fructose in the American diet. Abdominal (trunk) fat was measured via iDEXA (GE).[br]There were no significant changes in fasting glucose at any level of added sugar consumption (p[gt]0.05). Body mass, body fat and abdominal fat were unchanged in any group as were triglycerides (p[gt]0.05). Importantly none of these measure were influenced by the type or amount of added sugar consumed.[br]Sucrose and HFCS have comparable effects and do not cause metabolic abnormalities or increased abdominal fat when consumed at up to the 95th percentile population consumption level.[br][br]Sources of Research Support: Corn Refiners Association.[br][br]Nothing to Disclose: JL, ZY, SS, VN, SW, JR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 132 297 2059 MON-124 PO28-01 Monday 1727 2012


1724 ENDO12L_MON-125 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Peripheral Oxytocin Treatment Reduces Food Intake and Ameliorates Obesity, Fatty Liver and Glucose Intolerance in Diet-Induced Obese Mice Yuko Maejima, Yusaku Iwasaki, Takeshi Arai, Udval Sedbazar, Darambazar Gantulga, Toshihiko Yada Jichi Medical University School of Medicine, Shimotsuke, Japan; National Institute for Physiological Science, Okazaki, Japan Recent reports suggest a role of oxytocin (Oxt) in regulation of energy metabolism. We have examined acute and subchronic effects of Oxt on food intake and body weight (BW) in male and female mice.[br]Intraperitoneal (IP) injection of Oxt (400 [micro]g/kg BW) decreased food intake in male and female mice fed standard chow. IP injection of Oxt also decreased food intake in high fat diet-induced obese (DIO) male mice. Suppression of food intake by IP injection of Oxt was not altered by intracerebroventricular injection of a melanocortin receptor blocker, SHU9119. IP injection of Oxt increased c-FOS expression in several brain regions including paraventricular nucleus (PVN), arcuate nucleus (ARC), and nucleus tractus solitaries (NTS). These results suggested that IP Oxt injection induced anorexia via signaling to these brain regions and independently of melanocortin system.[br]Daily subcutaneous (SC) injection of Oxt (1,600 [micro]g/kg/day) for 17 days in male DIO mice reduced food intake for 6 days and BW for the entire treatment period and additional 9 days after terminating Oxt treatment.[br]Oxt infusion (1,600[sub] [micro][/sub]g/kg/day) by SC implanted osmotic mini pumps for 13 days in male and female DIO mice reduced food intake and BW. Oxt infusion also decreased visceral fat mass, adipocyte size, respiratory quotient in light phase, and ameliorated fatty liver and impaired glucose tolerance. Oxt infusion failed to affect normal blood pressure and locomotor activity observed in the male DIO mice. These metabolic effects of Oxt in male mice were also observed in female DIO mice but to a smaller extent, while the effect to improve glucose tolerance was greater in female than male DIO mice. Altogether, subchronic peripheral Oxt treatment reduced food intake and visceral fat mass, and ameliorated obesity, fatty liver and impaired glucose tolerance. Peripheral administration of Oxt provides a new therapeutic avenue for treatment of obesity, hyperphagia and type2 diabetes in both sexes.[br][br]Nothing to Disclose: YM, YI, TA, US, DG, TY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 135 297 2060 MON-125 PO28-01 Monday 1728 2012


1725 ENDO12L_MON-126 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Electron Microscopic Changes of Fatty Liver after Fenofibrate Treatment in OLETF Rats Mi-Kyoung Park, Ying Han, Su Kyoung Park, So-Young Park, Hyeongjong Koh, So Young Park, Duk Kyu Kim, Hye-Jeong Lee Dong-A University College of Medicine, Busan, Republic of Korea; Dong-A University College of Medicine, Busan, Republic of Korea The mitochondria in the hepatocyte oxidize fatty acids and generate energy. The deterioration of the mitochondrial function is understood as the important pathogenesis of obesity and diabetes. The fenofibrate induces proliferation of the peroxisomes and enhances fatty acid oxidation. Thus, we compared electron microscopic findings of the liver of obese rats before and after treatment of fenofibrate.[br]We used the Otsuka Long-Evans Tokushima Fatty(OLETF) rats(O) as the obese model (n=10) and the Long-Evans Tokushima(LETO) rats(L) as the control (n=10). Both of two models were divided to subgroup of the fenofibrate-treated(F) group and the control(C) group. All of four groups were fed liberally to 16 weeks of age. Since 17 weeks, the F group was fed with a standard chow and fenofibrate(30mg/kg/day) and C group was fed with a standard chow. All rats were fed chow food and water liberally. After 10 weeks of treatment, all of rats were sacrificed and the liver were gained and fixed immediately. We examined the hepatic mitochondria by transmission electron microscope.[br]Basal body weights at the age of 17 weeks were 587[plusmn]28g in O-C group (n=5), 587[plusmn]30g in O-F group (n=5), 455[plusmn]19g in L-C group, 457[plusmn]21g in L-F group. Final body weights at the age 28 weeks were 700[plusmn]33g in O-C group, 690[plusmn]36g in O-F group, 517[plusmn]31g in L-C group, 512[plusmn]30g in L-F group. Basal serum total cholesterol levels were 106[plusmn]16.4 mg/dL in L-C group, 95[plusmn]24.6mg/dL in L-F group, 299[plusmn]123.5mg/dL in O-C group, 172[plusmn]37.5mg/dL in O-F group. At the end of study, serum total cholesterol levels were 136[plusmn]20.8mg/dL in L-C group, 181[plusmn]36.5mg/dL in L-F group, 198[plusmn]22.9mg/dL in O-C group, 237[plusmn]36.6mg/dL in O-F group. Serum triglyceride levels were 40[plusmn]4.7mg/dL in L-C group, 43[plusmn]4.5mg/dL in L-F group, 93[plusmn]30.8mg/dL in O-C group, 105[plusmn]19.6mg/dL in O-F group at baseline and 50[plusmn]15.5mg/dL in L-C group, 44[plusmn]19.6mg/dL in L-F group, 247[plusmn]83.3mg/dL in O-C group, 298[plusmn]157.9mg/DL in O-F group at the end of the study. The hepatic mitochondria of the obese model showed destruction of spherical shape, swelling and inhomogeneity compared to the control. After 10 weeks of fenofibrate, the hepatic mitochondria were denser and compacter but the inhomogeneity was remained in O-F group compared with in O-C group. The peroxisomes were increased after fenofibrate therapy in O-F group.[br]In conclusion, in the obese model, the structure of hepatic mitochondria is deteriorated. The fenofibrate can reverse this change somewhat but not normalize.[br][br]Nothing to Disclose: M-KP, YH, SKP, S-YP, HK, SYP, DKK, H-JL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 741 297 2061 MON-126 PO28-01 Monday 1729 2012


1726 ENDO12L_MON-127 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Augmented Gastrointestinal Expression of Cytosolic Malic Enzyme (ME1) in Mice Pre-Disposes to Obesity Ahmed Al-Dwairi, Charles P Mercado, Hamdan Hamdan, John Mark P Pabona, Patricia A Wight, Rosalia CM Simmen, Frank A Simmen University of Arkansas for Medical Sciences, Little Rock, AR Cytosolic Malic Enzyme (ME1) converts malate to pyruvate, generating NADPH for de novo lipogenesis. Recently, adipose and liver ME1 has been correlated with susceptibility to obesity and type 2 diabetes. The role of intestinal ME1 in adiposity remains unexplored. We evaluated the effects of augmented ME1 expression in murine intestinal mucosa on body weight, adiposity, and serum hormone levels. Two lines of transgenic (Tg) mice were generated that express rat ME1 in the intestinal crypt and villus epithelium under the control of the murine villin promoter-enhancer. Intestinal expression of the rat ME1 transgene was confirmed by qRT-PCR, western blot, and immunohistochemistry. Male and female Tg [amp] wild type (WT) mice were fed a normal chow diet for 8 weeks beginning at weaning. Compared to WT males, Tg male mice had greater body weight at study termination (Tg line 1: 27.90 [plusmn] 0.41 g, P[lt]0.001 and Tg line 2: 25.88 [plusmn] 0.87 g; P=0.06 vs. WT: 23.71 [plusmn] 0.61 g). Liver weights in Tg line 1 differed from WT (1.48 [plusmn] 0.08 vs. 1.22 [plusmn] 0.04 g, P=0.016). Fat depot weights, and blood levels of glucose, insulin, leptin, gastric inhibitory peptide (GIP) and glucagon-like peptide 1 (GLP-1) did not differ between genotypes. However, expression of Proglucagon (precursor of GLP-1) in the ileums of Tg line 2 mice was reduced by [sim]50% (P=0.033) compared to WT. Colon crypts in males from Tg line 2 were deeper than those of corresponding WT mice (235.61 [plusmn] 8.47 vs. 200.58 [plusmn] 10.91 [micro]m, P=0.028). In contrast, no differences in body weight and hormone levels were apparent for WT and Tg females. To evaluate effects of augmented ME1 expression during diet-induced obesity, Tg and WT males were placed on a high fat diet for 15 weeks, post-weaning. Relative to WT mice, Tg line 1 had significantly increased gonadal fat pad weights (2.09 [plusmn] 0.10 g vs. 1.63 [plusmn] 0.17 g, P=0.007), and retroperitoneal fat pad weights (0.98 [plusmn] 0.06 g vs. 0.73 [plusmn] 0.11 g, P=0.073); although body weights did not differ. Analysis of serum hormones revealed an [sim]2-fold increase in GIP levels (P=0.006), and a tendency for an increase in leptin levels (P=0.06) in Tg line 1 compared to WT mice. Data highlight the importance of gastrointestinal ME1 as a determinant of obesity, and its potential as a target for interrupting the obesity-colon cancer connection. Trang Van and Letha McGarity provided valuable technical assistance.[br][br]Sources of Research Support: NIH R01CA136439, the Sturgis Foundation and the Arkansas Biosciences Institute-ACHRI.[br][br]Nothing to Disclose: AA-D, CPM, HH, JMPP, PAW, RCMS, FAS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 849 297 2062 MON-127 PO28-01 Monday 1730 2012


1727 ENDO12L_MON-128 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Thermogenesis in Adipose Tissue Explains the Body Composition of Genetically Lean and Obese Sheep Belinda A Henry, Ian R Young, Jonathan G Hickford, Iain J Clarke Monash University, Clayton, Australia; Lincoln University, Lincoln, New Zealand Body weight and adiposity are controlled by energy intake and energy expenditure. The latter comprises basal metabolic rate, physical activity and thermogenesis. Here, we sought to characterize the role of thermogenesis in sheep that have been selected through a number of generations to be either lean or obese (1). The cause of the lean or obese phenotype is unknown in these animals, for which body weight and food intake is similar (1). The aim of the present study was to measure post-prandial temperature in fat and muscle in order to ascertain if a difference in putative thermogenesis is the cause of the difference in body composition. Adult ewes (n=7 lean; n=8 obese) were studied in the non-breeding season, to avoid fluctuations in ovarian steroids. One lateral ventricle was cannulated and temperature sensing probes (Dataloggers) were implanted into the retroperitoneal fat and the skeletal muscle of the hind-limb. The dataloggers recorded temperature at 15min intervals. A rigid feeding regime (food available 1100-1600h) was imposed for 3 weeks prior to experimentation. Food intake was monitored throughout the experimental period. As previously reported (1), food intake was similar between lean and obese animals (lean: 20.6[plusmn]1.6 g/kg body weight vs obese: 16.7[plusmn]1.1 g/kg body weight). Lean and obese animals received either artificial cerebrospinal fluid or leptin (10[micro]g/h) from 0900-1500h. Following experimentation, body composition was measured by computerized tomography (CT) scan. Adiposity was greater (P[lt]0.001) in obese ewes (obese: 40.7[plusmn]2.6 vs lean: 18.3[plusmn]4.3 % adiposity). Prior to infusion, skeletal muscle temperature was similar in obese and lean animals, whereas fat temperature was higher (P[lt]0.01) in the lean group (obese: 0.86[plusmn]0.12 vs lean: 1.16[plusmn]0.1 [deg]C post-prandial change in temperature). Infusion of leptin increased (P[lt]0.05) temperature in fat and muscle tissues, but effects were similar between the lean and obese animals. Interestingly, leptin reduced food intake in obese animals only, indicating that these animals are not leptin-resistant. We conclude that innate differences in adiposity are due, at least in part, to differences in post-prandial thermogenesis in visceral fat.[br][br]1. Anukulkitch et al (2010) Neuroendocrinology 91: 223-238, 2010.[br][br]Nothing to Disclose: BAH, IRY, JGH, IJC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 853 297 2063 MON-128 PO28-01 Monday 1731 2012


1728 ENDO12L_MON-129 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Loss of Cytosolic Malic Enzyme (ME1) in Mice Confers an Endocrine Profile Favoring Decreased Risk of Obesity-Associated Colon Cancers Ahmed Al-Dwairi, Mark John P Pabona, Rosalia CM Simmen, Frank A Simmen University of Arkansas for Medical Sciences, Little Rock, AR Obesity and its associated hyperinsulinemia and hyperleptinemia are known risk factors for colon cancer. Cytosolic Malic Enzyme (ME1) generates NADPH for lipogenesis in liver and fat tissues, and is a major obesity and diabetes-associated gene. Soy protein based diets reduce ME1 gene expression in liver and fat; lower body fat content; and attenuate colon tumor incidence in rodent models. To evaluate the role of ME1 and its regulation by diet on endocrine and metabolic parameters implicated in obesity-associated colon cancers, we examined the effect of soy protein isolate (SPI)-supplemented diet on weight gain, hormonal status, and mRNA expression of lipogenic enzymes [ME1, Fatty Acid Synthase (FASN)] and proliferation-associated proteins (Ki-67, mTOR) in colons and small intestines of wild type (WT) and ME1 null (MOD-1 line) mice. Male WT and MOD-1 mice were weaned to a high fat (HF) diet containing casein (CAS-HF, control diet) or SPI (SPI-HF). Diets were fed for 5 wks. The SPI-HF-fed WT mice had a significant reduction in weight gain compared to the WT mice fed with CAS-HF (33.2 [plusmn] 0.79 vs. 29.3 [plusmn] 0.7 g, P[lt]0.001), whereas MOD-1 mice on both diets gained less weight (23.7 [plusmn] 0.61 g) than corresponding WT mice (P[lt]0.001). MOD-1 mice fed either diet had [sim]20% reduction in liver weight, [sim]50% reduction in gonadal and retroperitoneal fat weights (P[lt]0.01), and reduction in colon crypt depth (177 [plusmn] 9.28 vs. 141.3 [plusmn] 8.5 [micro]m, P[lt]0.01), compared to WT mice fed CAS-HF diet. SPI-HF-fed WT mice and MOD-1 mice fed either diet had 50-70% reductions in serum insulin and leptin levels (P[lt]0.001), [sim]50% reductions in colon mTOR and jejunum Ki-67 gene expression (P[lt]0.01), and [sim]25% reduction in jejunum FASN mRNA transcripts (P[lt]0.05), compared to CAS-HF-fed WT counterparts. However, colon and jejunum ME1 gene expression was unaffected by diet. SPI-HF diet increased serum adiponectin levels in WT mice. The leptin/adiponectin ratio was reduced by [sim]45% in SPI-HF-fed WT mice, and by [sim]75% in MOD-1 mice (both diets), when compared to WT mice fed CAS-HF (P[lt]0.001). Sera from MOD-1 mice decreased proliferation of HCT-116 human colon cancer cells, when compared to sera from WT mice (P[lt]0.05). The collective data suggest that the targeting of ME1, possibly by dietary factors with ME1-lowering activity such as SPI, may provide avenues to reduce obesity and its pro-tumorigenic endocrine environment, and thereby disrupt the obesity-colon cancer connection.[br][br]Sources of Research Support: SPI was provided by Solae, LLC; MOD-1 mice were from Merck [amp] Co., Inc.; supported by NIH R01CA136439, the Sturgis Foundation and the Arkansas Biosciences Institute-ACHRI.[br][br]Nothing to Disclose: AA-D, MJPP, RCMS, FAS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 854 297 2064 MON-129 PO28-01 Monday 1732 2012


1729 ENDO12L_MON-130 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Hyperprolactinemia Reduces Insulin Resistance and Prevents Visceral Adipose Tissue Hypertrophy in Obese Rats Xarubet Ruiz-Herrera, Maria Lemini, Fernando Lopez-Barrera, Edith Arnold, Gonzalo Martinez de la Escalera, Carmen Clapp, Yazmin Macotela Universidad Nacional Aut[oacute]noma de M[eacute]xico (UNAM), Quer[eacute]taro, Mexico Obesity is characterized by excessive accumulation of body fat. However, the metabolic impact of surplus fat depends on its localization. Increased storage of visceral adipose tissue is associated with bad metabolic outcomes, whereas an excess of subcutaneous adipose tissue can have neutral or even protective metabolic effects. Understanding how adipose tissue function is regulated within different depots is fundamental for developing effective therapies against obesity and its associated complications. Prolactin (PRL) is one factor that may regulate adipose tissue function and metabolic homeostasis. Human adipose tissue secretes PRL, and preadipocytes and adipocytes from mice visceral adipose tissue express higher levels of the PRL receptor compared to their subcutaneous counterparts. PRL can increase food intake in female rats, and reducing serum PRL with bromocriptine can result in weight loss in obese patients with prolactinomas. Also, PRL receptor knockout mice have reduced visceral adipose tissue mass compared to wild type animals. In the present work, we analyzed whether hyperprolactinemia (hyperPRL) influences insulin resistance and adipose tissue expansion in obese rats. Male Wistar rats were maintained with either a control diet (CD) or a high fat diet (HFD) and 4 weeks later, both groups were divided in half to form groups: CD, CD+hyperPRL, HFD and HFD+hyperPRL. Hyperprolactinemia was induced by placing subcutaneous osmotic pumps releasing PRL for 28 days, and PRL circulating levels were evaluated by the Nb2 bioassay. An insulin tolerance test was performed after 3 weeks of hyperPRL induction, and adipose tissues and serum were collected at the end of the following week. We found that hyperPRL significantly reduced HFD-induced insulin resistance and resulted in larger visceral and subcutaneous adipose depots in the HFD-fed animals. However, this was the result of increased hyperplasia in both subcutaneous and visceral fat but reduced hypertrophy specifically in visceral adipose tissue. HyperPRL had no effect on insulin sensitivity and adipose tissue expansion in CD-fed rats, or on food intake or body weight in rats under either CD or HFD. In conclusion, hyperprolactinemia could exert beneficial metabolic effects by modulating adipocyte growth and proliferation in obesity.[br][br]Sources of Research Support: Universidad Nacional Autonoma de Mexico (UNAM) (IB200411-22).[br][br]Nothing to Disclose: XR-H, ML, FL-B, EA, GMdlE, CC, YM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 865 297 2065 MON-130 PO28-01 Monday 1733 2012


1730 ENDO12L_MON-131 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Age at Onset of Diet-Induced Obesity Differentially Modifies the Metabolic Phenotype Observed in Adults Jose Cordoba-Chacon, Manuel David Gahete, Ana Isabel Pozo-Salas, Antonio Moreno-Herrera, Justo Pastor Castano, Rhonda Denise Kineman, Raul Miguel Luque University of C[oacute]rdoba, Instituto Maim[oacute]nides de Investigaci[oacute]n Biom[eacute]dica de C[oacute]rdoba (IMIBIC), and CIBER Fisiopatolog[iacute]a de la Obesidad y Nutrici[oacute]n, C[oacute]rdoba, Spain; University of Illinois at Chicago and Jesse Brown Veterans Affairs Medical Center, Research and Development Division, Chicago, IL Mice are commonly used as models to study the metabolic impact of diet-induced obesity (DIO). Many studies use high-fat (HF) diets to induce obesity, where the diet is introduced at 4 weeks of age, while others examine the impact of starting HF-feeding in adults. Since energy needs and sex hormone levels dramatically differ in the prepubertal vs. adult state, the current study was designed to determine if the age at onset of the obesity modifies the metabolic phenotype observed. To this end, male C57Bl/6J mice were fed a low-fat (LF, 10% kcal from fat) or HF (60% kcal from fat) diet starting during the prepubertal period (pDIO, 4 wks of age) or as adults (aDIO, 12 wks of age). When compared to LF-fed, age-matched controls, pDIO mice gained relatively more weight than aDIO mice, but absolute fat depot weights did not differ. Insulin levels were increased and glucose tolerance, as assessed by GTTs, was impaired by HF-feeding, but did not differ between pDIO (13 wks diet, 17 wks age) and aDIO (10 wks diet, 22 wks age). Interestingly, aDIO mice became more insulin resistant than pDIO mice, compared to their controls, as assessed by ITT performed 2 weeks following GTTs. The ability of pDIO mice to partially preserve insulin sensitivity, may be explained in part by an increase in both lean and fat mass, while only fat mass was increased in aDIO mice, as assessed by whole body NMR in separate sets of mice. Examination of circulating factors that may also contribute to these differences in insulin sensitivity revealed pDIO and aDIO displayed a rise in leptin, while circulating IGF-I and liver ALS mRNA were increased only in pDIO mice. A rise in IGF-I has been previously been reported in the pDIO model, where IGF-I may serve to improve insulin sensitivity. HF-feeding increased corticosterone levels, which reached significance in pDIO mice. Intriguingly, hypothalamic expression of CRF was reduced in pDIO mice, without changes in circulating ACTH, whereas aDIO mice displayed a clear increase in ACTH, without alterations in CRF expression, thus suggesting that aDIO mice may have defects in negative feedback regulation of adrenal-axis function. Taken together, these initial observations clearly demonstrate that the age at onset of obesity should be considered when designing studies to examine metabolic function in different physiologic/genomic states since it may produce important and specific changes in body composition, adrenal-axis function and insulin sensitivity.[br][br]Sources of Research Support: This work was supported by the following grants: Fundaci[oacute]n Alfonso Martin Escudero (J.C-C), Fundaci[oacute]n Caja Madrid (M.D.G.), Department of Veterans Affairs, NIH R01DK088133 (R.D.K.) BFU2010-19300, RYC-2007-00186, CIBERObn (MICINN/FEDER), BIO-139, CTS-5051 (Junta de Andaluc[iacute]a) (J.P.C; R.L.M).[br][br]Nothing to Disclose: JC-C, MDG, AIP-S, AM-H, JPC, RDK, RML 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1049 297 2066 MON-131 PO28-01 Monday 1734 2012


1731 ENDO12L_MON-132 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Postnatal Early Overnutrition Programs for Higher Liver Oxidative Stress, Liver Steatosis and Impairment of Insulin Signaling Pathway in the Adult Rat Offspring Juliana Gastao Franco, Ellen Paula Conceicao, Elaine Oliveira, Nayara Peixoto-Silva, Patricia Cristina Lisboa, Egberto Gaspar Moura State University of Rio de Janeiro, Rio de Janeiro, Brazil Postnatal early overfeeding (EO) is related to later development of overweight and other metabolic disorders (1,2). As oxidative stress is implicated in most human diseases, as obesity and diabetes, we decided to study some parameters related to oxidative stress and insulin signaling in liver from EO animals in adult life. To induce EO, litter size was reduced to 3 pups/litter (SL: small litter) and groups with normal litter size (NL:10 pups/litter) were used as control, at the third postnatal day. At least, 6 litters were studied per group. After weaning, rats had free access to standard diet and water. Body weight and food intake were monitored daily and male offspring were killed at 180 days-old. Significant differences had p[lt]0.05 or less. As expected, SL rats had hyperphagia, higher body weight and higher visceral fat mass at weaning and adulthood. At adulthood, postnatal EO programmed for lower catalase (-42%), superoxide dismutase (-45%) and glutathione peroxidase (-65%) activities in liver. The evaluation of liver injury in adult SL group showed lower nitrite content (-10%), higher liver and plasma malondialdehyde content (+25% and 1.1 fold-increase, respectively) that measures lipid peroxidation. No changes of total protein bound carbonyl or Cu/Zn SOD protein expression in liver were detected between the groups. Regarding insulin signaling pathway in liver, SL offspring showed lower IR[beta] (-66%), IRS1 (-50%), phospho-IRS1 (-73%), PI3-K (-30%) and Akt1 (-58%). Indeed, morphological analysis showed that SL rats presented focal areas of inflammatory cell infiltrate and lipid drops in their cytoplasm characterizing a microsteatosis. Thus, we evidenced that postnatal EO can program the oxidative stress in liver, maybe contributing for impairment of insulin signaling and non alcoholic liver steatosis.[br][br]1. Rodrigues AL et al, J Nutr Biochem 2011;22:109. 2. Conceicao EP et al, Horm Metab Res 2011;43:513.[br][br]Sources of Research Support: National Council for Scientific and Technological Development (CNPq), Coordination for the Enhancement of Higher Education Personnel (CAPES), and State of Rio de Janeiro Carlos Chagas Filho Research Foundation (FAPERJ).[br][br]Nothing to Disclose: JGF, EPC, EO, NP-S, PCL, EGM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1110 297 2067 MON-132 PO28-01 Monday 1735 2012


1732 ENDO12L_MON-133 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) An Obesity-Dependent Lactation Defect in the Viable Yellow Agouti Mouse Is Associated with Mammary Inflammation Darryl L Hadsell, Louise A Hadsell, Walter Olea Baylor College, Houston, TX Maternal obesity is known to delay lactogenesis in breast feeding women as well as negatively impact lactation in other species. Obesity is also understood to be associated with inflammation. Work with the viable yellow agouti (A[sup]vy[/sup]) mouse in our laboratory has documented a lactation defect in obese yellow A[sup]vy[/sup] females. The hypothesis for this study was that increased mammary infiltration of inflammatory macrophages is present with the obesity- dependent lactation defect in A[sup]vy[/sup] mice. At weaning, female a/A[sup]vy[/sup] mice (obese) and their a/a littermates (lean) were allowed ad-libtum access to standard chow until Obese mice reach over 40% body fat at mating. A cohort of a/A[sup]vy[/sup] mice was diet restricted (CR) to match the weight of Lean littermates, both of which had less than 16% body fat. At day 1 or 4 postpartum, dams were killed and mammary glands (MG) were harvested. Litter gain and pup survival was lower (P[lt]0.05) in Obese than Lean dams. This defect was rescued by preventing obesity in a/A[sup]vy[/sup] females with food restriction. Immunostaining for the macrophage marker F4/80 demonstrated that there was both an increase in macrophages and an increase in the presence of crown-like structures in mammary glands of obese mice. Double-labeling for F4/80 and iNOS showed that the percentage of iNOS-positive macrophages was higher in mammary tissue of obese dams. Real-time PCR on total mammary tissue RNA demonstrated that the mRNA for Lalba and Csn2 were decreased in obese dams while those for F4/80, Ccl2, and Tnf[alpha] were increased. These changes were prevented by preventing obesity through food-restriction. These results support the conclusion that the lactation defect and the presence of mammary inflammation in the A[sup]vy[/sup] mouse are due to maternal obesity.[br][br]Sources of Research Support: This project was funded by a grant from NICHD # 1 R21 HD056439-01A2/02.[br][br]Nothing to Disclose: DLH, LAH, WO 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1424 297 2068 MON-133 PO28-01 Monday 1736 2012


1733 ENDO12L_MON-134 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Strain-Related Differences in Response to Diet-Induced Obesity and PPAR-[gamma] Agonist Treatment Emily K Sims, Masayuki Hatanaka, Sarah A Tersey, Tatsuyoshi Kono, Dan R Moss, Natalie D Stull, Raghavendra G Mirmira, Carmella Evans-Molina Indiana University School of Medicine, Indianapolis, IN; Indiana University School of Medicine, Indianapolis, IN; Indiana University School of Medicine, Indianapolis, IN Previous literature has shown that C57BL/6J (BL/6J) mice exhibit greater susceptibility to diet-induced obesity (DIO) compared to C57BL/KsJ (BL/KsJ) mice. In the context of leptin deficiency, both strains develop obesity, yet glucose homeostasis and pancreatic islet function are significantly worse in BL/KsJ mice. We hypothesized that these distinct responses to DIO would be informative in understanding diabetes susceptibility and response to pharmacological therapies. To this end, BL/6J and BL/KsJ mice were fed normal chow (NC) containing 18% of calories from fat, high fat diet (HFD) with 42% of calories from fat, or HFD with 20mg/kg/day of pioglitazone (HFD+Pio) for a 12-wk period, beginning at 8 wks of age. At baseline, BL/KsJ mice had lower body weight, lean mass (assessed by dual-energy X-ray absorptiometry) and glucose tolerance compared to BL/6J mice. Percentage of body fat (%BF) and insulin sensitivity were similar among the groups. With HFD, BL/6J mice showed greater weight gain. Compared to HFD alone, HFD+Pio did not impact body weight or composition further, yet caloric intake was higher in this group. By contrast, BL/KsJ mice fed HFD had no significant increase in weight compared to NC, yet exhibited a significant increase in %BF. BL/KsJ mice fed HFD manifested a significant reduction in food and caloric consumption, and demonstrated increased movement. Interestingly, compared to NC and HFD, HFD+Pio significantly increased weight gain in BL/KsJ mice with a trend towards increased %BF. Respiratory quotients were measured by indirect calorimetry and found to be significantly decreased with HFD in both strains and increased with the addition of Pio. Results from glucose tolerance tests showed a significant increase in the area under the curve in BL/6J mice fed HFD. The addition of Pio significantly improved glucose tolerance and insulin sensitivity to levels indistinguishable from NC. Similar trends in glucose tolerance were observed in the BL/KsJ strain but differences did not reach statistical significance. In summary, meaningful metabolic differences were observed between BL/6J and BL/KsJ mice in response to HFD and Pio treatment. BL/KsJ were resistant to weight gain with HFD, which may in part be explained by effects on food intake and compensatory energy expenditure. Mechanistic islet, liver, and adipose-focused studies are ongoing with the goal of understanding differences in the responses of human populations to a typical Western diet.[br][br]Nothing to Disclose: EKS, MH, SAT, TK, DRM, NDS, RGM, CE-M 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1680 297 2069 MON-134 PO28-01 Monday 1737 2012


1734 ENDO12L_MON-135 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Transient Glucocorticoid Excess Causes Persisting Changes in Lean Body Mass and Fat Mass in Mice Fed a High-Fat Diet Hanna E Auvinen, Johannes A Romijn, Nienke R Biermasz, Mariette R Boon, Louis M Havekes, Johannes WA Smit, Patrick CN Rensen, Alberto M Pereira Leiden University Medical Center, Leiden, Netherlands Introduction In humans, Cushing[apos]s syndrome (CS) is associated with an increased incidence of the metabolic syndrome, increased cardiovascular morbidity and mortality, even after long-term correction of glucocorticoid (GC) excess.[br]Aim To evaluate the effects of transient overexposure to GC on the metabolic changes in the long-term in mice.[br]Methods Single housed male C57Bl/6J mice were fed either a low fat (LFD) or high fat diet (HFD) and given corticosterone (CORT) (50 microg/ml) or vehicle in the drinking water for 4 wks, followed by a washout period for 4 or 8 wks thereafter. Plasma circadian corticosterone levels were assessed at baseline and at wks 4, 8, and 12 after the start of exposure. Lipids, insulin, and glucose levels were measured after an overnight fast. Insulin sensitivity was assessed by hyperinsulenemic-euglycemic clamp at week 8 and 12 for the low fat diet fed mice and week 12 for the high fat diet fed mice. Lean and body- and fat mass were analyzed by DEXA.[br]Results CORT-treatment transiently increased plasma corticosterone by 37-, 13-, 3- and 13-fold (LFD), and 25-, 5-, 2-, and 9-fold (HFD) at 07.00h, 12.00h, 18.00h and 22.00h, respectively. At week 8, peak CORT levels at 18.00h were suppressed in both diet groups, returning to baseline levels at week 12. CORT-treatment increased food intake and plasma levels of insulin, trigycerides, free fatty acids and cholesterol in both diet groups. Abrogation of CORT normalized food intake, whereas body weight was increased in HFD fed mice but remained unchanged in LFD fed mice. At week 12, insulin was still significantly higher in CORT treated mice in both diet groups. There were no differences in the lean body or fat mass at week 8 and 12 weeks in LFD fed mice but HFD fed CORT mice had persistently lower lean body mass and higher fat mass. Hyperinsulinemic-euglycemic clamp at week 8 or 12 in CORT-treated mice vs controls indicated no changes in either diet group.[br]Conclusion In mice, transient CORT excess induces long-lasting changes in body composition only in the presence of HFD. These diet dependent effects of CORT might contribute to the persistent adverse cardiovascular risk profile as observed in patients treated for Cushing[apos]s syndrome.[br][br]Nothing to Disclose: HEA, JAR, NRB, MRB, LMH, JWAS, PCNR, AMP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2142 297 2070 MON-135 PO28-01 Monday 1738 2012


1735 ENDO12L_MON-136 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Autophagy Reverses Intracellular Lipid Accumulation in Skeletal Muscle Truong N Lam, Husnu E Kaynak, Mary Lim, Romain Harmancey, Heinrich Taegtmeyer University of Texas Medical School at Houston, Houston, TX Autophagy is a tightly regulated cellular housekeeping process involving the degradation and recycling of protein aggregates, or damaged cell organelles. Autophagy may also contribute to the mobilization of intracellular lipid stores ([quot]lipophagy[quot]). We previously reported that bariatric surgery leads to a dramatic reversal of triglyceride accumulation in skeletal muscle of morbidly obese patients (1, 2). We now proposed that the process of lipid clearance from muscle cells which accompanies weight loss surgery is linked to an increase activity of autophagy. In order to test our hypothesis, muscle biopsy samples were recovered from the vastus lateralis of 5 severely obese patients (BMI=46.7[plusmn]1.7 kg/m[sup]2[/sup]) before and 9 months after bariatric surgery. Triglycerides were extracted from biopsies and quantified using an enzymatic assay, and the expression level of autophagy markers (p62, LC3, Beclin1) was assessed by immunoblotting. In order to confirm a role for autophagy in lipid clearance from muscle cells, L6 rat myocytes were incubated for 5 days in presence of fatty acids (0.5 mM or 1 mM). The cells were then treated for 24 h with an inhibitor (200 nM bafilomycin A1) or an activator (1 [mu]M rapamycin) of autophagy, and the effects on intracellular triglyceride accumulation assessed by Oil Red O staining and enzymatic quantification. Among the 5 morbidly obese patients, 3 presented a reduction in intramyocellular triglyceride levels 9 months after bariatric surgery, while the 2 others displayed increased triglyceride levels. The expression of the polyubiquitin-binding protein p62 was inversely correlated to the changes in intramuscular triglyceride levels, suggesting that increased autophagic activity correlates with a reduction in lipid levels. In L6 myocytes, autophagy inhibition increased lipid accumulation and cell death, while the activation of the process was associated with a lower accumulation of triglycerides. In conclusion, increased autophagic activity is related to an increased clearance of lipids from muscle cells. We propose that increased lipophagy may contribute to the reversal of lipotoxic mechanisms in skeletal muscle of severely obese patients.[br][br](1) Leichman JG et al., Am J Med 2008; 121:966. (2) Algahim MF et al., Am J Physiol Heart Circ Physiol 2012; Epub ahead of print.[br][br]Nothing to Disclose: TNL, HEK, ML, RH, HT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2191 297 2071 MON-136 PO28-01 Monday 1739 2012


1736 ENDO12L_MON-137 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Metabolic Factors That Lead to Obesity in Mice with Complete Deletion of Somatotrope Leptin Receptors (Somatotrope Null LEPR Exon 1) Melody Lyn Allensworth-James, Angela Katherine Odle, Anessa C Haney, Gwen Vaughn Childs University of Arkansas for Medical Sciences, Little Rock, AR Leptin receptor signaling has been shown to be important for appetite control, energy expenditure, and development. Our lab is interested in the role of leptin in pituitary somatotrope regulation. Mice with somatotrope-specific deletion of exon 17 of the LEPRb isoform, which encodes the JAK binding site, grow normally but are growth hormone deficient (GHD) and obese as adults (Childs et al, 2011). The present study tested mice with somatotrope-specific deletion of exon 1 of LEPR to determine if loss of all isoforms of the receptor (Cohen et al, 2001) would produce a more significant phenotype. Homozygous floxed exon 1 LEPR mice (J. Friedman) were crossed with mice bearing Cre-recombinase driven by rGH promoter (R. Kineman; Luque et al, 2007). Organ genotyping showed no evidence of extra-pituitary Cre-recombinase. Somatotrope LEPR Exon 1-null deletion mutants grew normally but showed evidence of increased abdominal fat mass by MRI as early as 2 months, before they exhibited increased body weight. Percentages of immunolabeled growth hormone cells were reduced by 34.2% in deletion mutants. To evaluate the metabolic status of the mice before they became obese, mutant and littermate control male mice were placed in a Comprehensive Laboratory Animal Monitoring System (CLAMS; Oxymax, Columbus Instruments). At 2.5-4 months of age, pre-obese deletion mutant males had significantly higher respiratory quotients (p[lt].0342) and were less active in both dark (19% lower, p[lt].006) and light (16% lower, p[lt].017) cycles. Mutants ate 21% more food/feeding bout than controls during the dark cycle (p[lt].003), but not more food overall. Serum leptin was within the normal range for control mice. Sleep analyses showed that mutants sleep 30% less overall (p[lt].032) with no difference in the dark phase but 37% lower in the light phase (p[lt].019). In addition, serum assays (Millipore) showed 25% increases in adiponectin (p[lt].048) which is produced by adipocytes, and is a sign of adipogenesis. In addition, the lipogenic protein, glucose dependent insulinotropic peptide (GIP) was significantly increased (by 57%, p[lt].032). The proinflammatory cytokine, interleukin-6 (44%, p[lt].024) was lower, which correlates with higher adiponectin. The enhanced carbohydrate oxidation (RQ), lower activity and disrupted sleep are primary contributors to the increased fat mass. The findings indicate the importance of leptin signaling to somatotropes as they optimize body composition.[br][br]Childs GV et al., Endocrinology 2011; 152:69. Cohen P, et al., J Clin Invest 2001; 108:1113. Luque RM, Endocrinology 2007; 148:1946.[br][br]Sources of Research Support: 1R01HD059056 awarded to GVC; NCRR P20 RR020146; P30. NS047546 core facilities.[br][br]Nothing to Disclose: MLA-J, AKO, ACH, GVC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2198 297 2072 MON-137 PO28-01 Monday 1740 2012


1737 ENDO12L_MON-138 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Circadian Control of [italic]Leptin[/italic] Expression and Signaling Maintains Energy Homeostasis [italic]In Vivo[/italic] Xiaoqian Fang, Nicole M Kettner, Derek K Bolinger, David D Moore, Loning Fu Baylor College of Medicine, Houston, TX Circadian rhythms are biological rhythms that control most aspects of mammalian physiology. These rhythms are generated by an endogenous mechanism called the circadian clock. We have demonstrated that disruption of circadian rhythm abolishes energy homeostasis and induces obesity in mice independent of all previous identified risk factors for obesity, and that the expression of the key peripheral adiposity signal leptin is primarily controlled by the molecular clock in the fat tissue but not exogenous food cues. Disruption of circadian homeostasis in mice by circadian gene mutation or jet-lag abolishes the circadian homeostasis of leptin expression and induces Leptin resistance in mice, a key pathophysiological mechanism for human obesity. Using in vivo ChIP approach, we demonstrated that the molecular clock directly control the transcription of leptin in the fat tissue by rhythmically competing with other transcription factors to bind to the same promoter region on the leptin promoter. We propose that disruption of circadian control of Leptin expression and signaling is a major contributor to the diverse metabolic consequences of circadian disruption.[br][br]Sources of Research Support: USDA/ARS Grant 6250-51000-055 awarded to LFU. NIH Grant R01 CA137019-01A awarded to LFU.[br][br]Nothing to Disclose: XF, NMK, DKB, DDM, LF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2339 297 2073 MON-138 PO28-01 Monday 1741 2012


1738 ENDO12L_MON-139 POSTER SESSION: Obesity Topics I (1:30 PM-3:30 PM) Ghrelin Receptor[ndash]Deficient Rats Resist Diet-Induced Obesity Valerie Charbonneau, Harry MacKay, Alfonso Abizaid Carleton University, Ottawa, Canada Ghrelin, an orexigenic hormone peptide, is the endogenous ligand for the growth hormone secretagogue receptor (GHSR). Previous research has established ghrelin as a modulator in the regulation of appetitive behaviors and energy balance, and this research has included the use of genetically altered mice with mutations to either the ghrelin or the ghrelin receptor (GHSR) gene. Recently a rat model with a mutation to the GHSR gene has become available and the present project attempted to validate this GHSR KO rat model. To this end, GHRS KO rats and their wildtype (WT) littermates (n= 8/group) originally obtained from Transposagen Bio (Lexington, KY) were single housed and monitored for food intake and bodyweight for a period of 5 months. Metabolic parameters were examined and compared with wild type littermates. During the first month animals had access to standard chow, whereas the remaining four months the animals were exposed to a High fat diet (60% calories from fat). Our results show that when correcting for body weight, GHSR KO rats ate less than their WT littermates. This was observed regardless of the diet the animals consumed and in spite of body weight gain not being different between the two strains. Interestingly, and after four months of being fed a high fat diet, KO rats had slightly lower fasting glucose levels compared to WT controls and exhibited more efficient glucose clearance following a glucose challenge. As a whole, our data reinforces the hypothesis that the ghrelin receptor is a potential target to prevent diet induced obesity, and provide for the GHSR KO rat as a valid model for the study of ghrelin biology.[br][br]Nothing to Disclose: VC, HM, AA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 423 297 2074 MON-139 PO28-01 Monday 1742 2012


1739 ENDO12L_MON-154 POSTER SESSION: Vascular Biology (1:30 PM-3:30 PM) Leptin Induces Activation and Inflammation of Rat and Human Chemoreceptors in the Carotid Body and Brainstem Astrocytes Nephtali Marina, Vitaliy Kasymov, David Braude, Michael P Gilbey, Alexander V Gourine, Vidya Mohamed-Ali University College London, London, UK; University College London, London, UK BACKGROUND: Chronic hyperleptinemia in obesity is often associated with persistent elevation in muscle vasoconstrictor sympathetic activity which may contribute to the development of obesity-related hypertension. Activation of peripheral and central chemoreceptors in the carotid bodies and brainstem astroglial cells is well known to produce reflex increases in sympathetic tone and blood pressure (chemoreflex). Recent studies have shown that leptin and leptin receptors are normally expressed both in carotid body Type I cells (1) and in astroglial cells (2). We hypothesize that exaggerated chemoreflex activation induced by leptin may underlie increases in sympathetic tone and blood pressure in obesity. METHODS: To explore the above hypothesis we investigated the functionality of these receptors by measuring intracellular calcium responses to leptin in rat carotid body dissociated cultures and in primary cultured rat and human brainstem astrocytes loaded with Ca[sup]2+[/sup] indicator Fura-2. Since inflammation has been implicated in the pathogenesis of obesity-related hypertension, we used immunofluorescence to determine whether chronic leptin treatment is associated with increased levels of pro-inflammatory cytokines and their receptors in cultured rat carotid body Type I cells and brainstem astrocytes. RESULTS: Exogenous application of recombinant rat or human leptin evoked dose-dependent calcium responses in individual carotid body Type I cells and in rat and human brainstem astrocytes, respectively. Typically, [Ca[sup]2+[/sup]][sub]i[/sub] responses had long latency (approximately 500 sec.) and were characterized by sustained [Ca[sup]2+[/sup]][sub]i[/sub] oscillations (maximum duration of approximately 20 min). Immunocytochemical analysis revealed that chronic leptin treatment (24 hrs) significantly increased expression of Interleukin 1-[beta] (IL1-[beta]), tumor necrosis factor (TNF[alpha]) and IL1-Receptor I in carotid body Type I cells and induced a proinflammatory profile in human astrocytes (P[lt]0.05 vs control). CONCLUSIONS: These data suggest that leptin induces activation and inflammation of peripheral and central chemoreceptors in the carotid bodies and in the brainstem, respectively. The ability of chemoreceptors of the carotid body and brainstem to respond to changes in systemic adipokines may explain, to some extent, the association between obesity and hypertension.[br][br](1) Porzionato A et al., Brain Res 2011; 1385:56. (2) Hsuchou H et al., Brain 2009;132t 4):889.[br][br]Sources of Research Support: NM is a British Heart Foundation Intermediate Basic Science Research Fellow. AV is a Wellcome Trust Senior Research Fellow.[br][br]Nothing to Disclose: NM, VK, DB, MPG, AVG, VM-A 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2037 298 2075 MON-154 PO10-02 Monday 1743 2012


1740 ENDO12L_MON-155 POSTER SESSION: Vascular Biology (1:30 PM-3:30 PM) Oral Relaxin (RLX) Improves Functional and Cognitive Recovery in Post- Stroke Patients Paolo Milia, Marco Caserio, Bernardo Bigazzi, Daniele Bani, Mario Bigazzi, Tito Rastelli, Francesco Sonaglia, Benedetta Bigazzi Prosperius Institute, Umbertide, Italy; Prospetius Institute, Umbertide, Italy; Prosperius Institute, Florence, Italy; University of Florence, Florence, Italy; Prosperius Institute, Florence, Italy; Prosperius Tiberino Institute, Umbertide, Italy; Prosperius Tiberino Institute, Umbertide, Italy; Prosperius Institute, Florence, Italy [bold]Introduction: [/bold]Serious psyco-neurological disabilities are common consequences in stroke survivors. RLX, through NO synthesis, has protective effects on experimental ischemia in the heart (1) and brain (2) and can improve brain functions (3). Aim of the present trial is to evaluate the effects of RLX in the recovery of post-stroke patients.[br][bold]Materials/Methods: [/bold]36 post-stroke patients admitted to our Rehabilitation Unit, were randomly assigned to either study cohorts: 18 patients (age 64-79, M 56%) received rehabilitation and porcine RLX (Vitalaxin Plus[sup]TM[/sup],Sky BioHealth, USA), 40 mg/day (group RLX+R); 18 patients (age 64-78, M 50%) only underwent rehabilitation (group R). The trial was approved by the local ethical Committee. Patients, or legally accepted surrogates, provided informed consent prior to entry the study. There were no differences between the two groups in terms of risk factors, stroke syndromes and etiology. Analysis were done at admission, (T0), 20 (T1), and 40 (T2) days by using: Functional Independent Measure (FIM) for daily activity; Trail Making Test (TMT) for cognitive function; and modified Ranking Scale (mRS) for global function. We compared groups for categorical variables, with the chi-square test with Yate[apos]s correction or Fisher exact test when appropriate, and for continuous variables with the Mann and Withney U test, using the SPSS statistical analysis software.[br][bold]Results[/bold]: During the trial, the patients of the RLX+R group showed better general conditions and resistance to afford rehabilitation exercises. FIM score after 20 days (T1) showed no significant differences between the groups (78 vs. 69; n.s.), while after 40 days (T2) the patients of RLX+R group showed a better recovery (96 vs. 75; [italic]p[/italic][lt]0.001). Cognitive function, assayed by TMT score, was also improved: the RLX+R patients revealed a better performance at T1 (3.5 vs. 2; [italic]p[/italic][lt] 0.002) and even more at T2 (4 vs. 2; [italic]p[/italic][lt] 0.001). Accordingly, global function, assayed by mRS score, was improved in the RLX+R patients at both T1 (2.5 vs. 3; [italic]p[/italic][lt]0.001) and T2 (2 vs. 3; [italic]p[/italic][lt]0.001). RLX did not cause any negative side effects. [bold]Conclusions:[/bold] Treatment with RLX resulted very safe and efficacious, as it clearly improved the psyco-neurological recovery of post-stroke patients as well as their general conditions. We may speculate that RLX hormone could represent a new therapeutic and preventative drug to reduce the incidence and disability of neurovascular ischemic diseases.[br][br]1) Masini E., Bani D., Bello MG., Bigazzi M., Mannaioni PF., Bani Sacchi T. Relaxin counteracts myocardial damage induced by ischemia-reperfusion in isolated guinea pig hearts: evidence for an involvment of nitric oxide. Endocrinology, 138:4713-9, 1997. 2) Wilson BC, Connell B, Saleh TM. Relaxin-induced reduction of infarct size in male rats receiving MCAO is dependent on nitric oxide synthesis and not estrogenic mechanisms. Neurosci Lett, 393:160-4, 2006. 3) Smith CM, Ryan PJ, Hosken IT, Ma S, Gundlach AL. Relaxin-3 systems in the brain--the first 10 years. J Chem Neuroanat, 42:262-75, 2011. 4) Bigazzi M, Bani D, Bani Sacchi T. Relaxin: a possible future preventative therapy for cardiovascular disease in postmenopausal women and men? Climacteric, 4:137-43, 2001.[br][br]Nothing to Disclose: PM, MC, BB, DB, MB, TR, FS, BB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 475 298 2076 MON-155 PO10-02 Monday 1744 2012


1741 ENDO12L_MON-156 POSTER SESSION: Vascular Biology (1:30 PM-3:30 PM) IGFBP-5 Mediates High-Glucose Induced Cell Proliferation and Collagen Synthesis in Cardiac Fibroblast Seung Eun Song, Yong-Woon Kim, Jong-Yeon Kim, Kyu-Chang Won, Jae-Ryong Kim, So-Young Park College of Medicine, Yeungnam University, Daegu, Republic of Korea; College of Medicine, Yeungnam University, Daegu, Republic of Korea; College of Medicine, Yeungnam University, Daegu, Republic of Korea; College of Medicine, Yeungnam University, Daegu, Republic of Korea Hyperglycemia promotes the progression of heart disease mainly by excessive interstitial myocardial collagen accumulation, which can result in impaired diastolic and systolic function. Insulin-like growth factor-binding protein (IGFBP-5) induces production and deposition of collagen and fibronectin in adult lung fibroblasts. In addition, IGFBP-5 expression is associated with idiopathic pulmonary fibrosis. The present study examined the effect of high glucose on IGFBP-5 expression and whether IGFBP-5 mediates high glucose-induced collagen synthesis in heart. Cardiac fibroblasts were isolated from the heart of Sprague-Dawley young rats (1 to 3 day-old) and passages 2-3 were used for this experiment. High glucose increased cell proliferation in a dose dependent manner at 48 h. The mRNA expression of MMP2, MMP9, collagen III was significantly increased by high glucose in a time dependent manner. Increased cell number by high glucose was significantly inhibited by ERK inhibitors (PD98059). The mRNA expression and protein level of IGFBP-5 was significantly increased by high glucose in a dose- and time-dependent manner in cardiac fibroblast. Increased IGFBP-5 expression by high glucose was inhibited by PD98059. In addition, rmIGFBP5 increased cell proliferation in a dose- and time-dependent manner. The mRNA expression of collagen III was significantly increased by rmIGFBP5 in a time dependent manner. Increased cell proliferation by rmIGFBP5 was significantly inhibited by IGFBP-5 siRNA with high glucose at 48 h. IGFBP-5 was also increased in the heart of OLETF rats. These results suggest that high glucose-induced cell proliferation and production of collagen III is mediated by IGFBP-5 in cardiac fibroblast.[br][br]Sources of Research Support: This work was supported by the Korea Science and. Engineering Foundation grant (2010-0007389) funded by. the Korea government.[br][br]Nothing to Disclose: SES, Y-WK, J-YK, K-CW, J-RK, S-YP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 727 298 2077 MON-156 PO10-02 Monday 1745 2012


1742 ENDO12L_MON-157 POSTER SESSION: Vascular Biology (1:30 PM-3:30 PM) Water-Soluble Danshen Extracts Have Therapeutic Effects on Atherosclerosis Cheol Ryong Ku, Yoon Hee Cho, Zhen-Yu Hong, Miran Seo, Eun Jig Lee Severance Hospital Integrative Research Institute for Cerebral [amp] Cardiovascular Diseases, Yonsei University College of Medicine, Seoul, Republic of Korea; Northwestern University, Feinberg School of Medicine, Chicago, IL [bold][italic]Objective[/italic]: [/bold]Danshen, dried root of [italic]Salvia miltiorrhiza[/italic], is one kind of traditional Chinese medicine that has many effects on cardiovascular diseases. This study aimed to evaluate the mechanisms that involve the anti-atherogenic effect of Danshen extracts (DE), which has especially abundant water soluble compounds.[br][bold][italic]Research Design and Methods[/italic]: [/bold]With HPLC/time-of-flight mass spectrometry (HPLC-TOF-MS), the compositions of DE were analyzed. Rat vascular smooth muscle cells (VSMCs) and human umbilical vein endothelial cells (HUVECs) were treated with DE. The effect of DE on cell proliferation, wound healing, cell cycle, and reactive oxygen species were evaluated. For evaluation of DE effect on [italic]in vivo[/italic] condition, carotid balloon injury and tail vein thrombosis were induced to Sprague-Dawley (SD) rats. Inhibitory action of DE on platelet aggregation was confirmed by impedance aggregometer. At last, the effect of DE as an anti-atherogenic therapeutics was evaluated with the rabbit iliac artery stent model.[br][bold][italic]Results[/italic]: [/bold]HPLC-TOF-MS revealed that DE had prominent water-soluble components with insignificant lipid-soluble ones. DE inhibited the production of reactive oxygen species, migration, and proliferation of PDGF-BB stimulated VSMCs. Furthermore, DE prevented HUVECs from inflammation and apoptosis. DE prevented TNF-[alpha]-induced MAPK and NF-[kappa]B activations in HUVECs, leading to down-regulating VCAM-1. Both effects of DE were reconfirmed in carotid balloon injured rat model. DE treatment attenuated the platelet aggregation in [italic]in vivo[/italic] and [italic]ex vivo[/italic] conditions, confirmed by impedance aggregometer. Pretreatment of DE prevented tail vein thrombosis, induced by [kappa]-carrageenan. Finally, DE treated rabbits showed decreased in-stent restenosis in stented iliac arteries.[br][bold][italic]Conclusions[/italic]: [/bold]Our study demonstrats that DE modulates the key atherogenic events in VSMCs, endothelial cells, and platelets both on [italic]in vitro[/italic] and [italic]in vivo[/italic] conditions.[br][br]Nothing to Disclose: CRK, YC, Z-YH, MS, EJL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 346 298 2078 MON-157 PO10-02 Monday 1746 2012


1743 ENDO12L_MON-158 POSTER SESSION: Vascular Biology (1:30 PM-3:30 PM) Soluble Receptor Activator of Nuclear Factor kappa B (sRANKL) Is Decreased in HIV-Infected Individuals with Increased Coronary Plaque Burden and Coronary Artery Calcium Janice Jin Hwang, Jeffrey Wei, Suhny Abbara, Steven Grinspoon, Janet Lo Massachusetts General Hospital, Boston, MA; Massachusetts General Hospital, Boston, MA Objective: There is an increased prevalence of coronary artery disease amongst patients with HIV. Receptor activator of nuclear factor kappa B (RANKL) and its decoy receptor, osteoprotegerin (OPG), have been implicated as potential mediators of vascular calcification. We sought to investigate the relationship between coronary artery calcification as measured by coronary computed tomography and soluble RANKL (sRANKL) and OPG in asymptomatic HIV-infected men and controls.[br]Methods: 78 HIV-infected men (age 46.5 [plusmn] 6.5 years, duration of HIV 13.5[plusmn]6.1 years, CD4 T-lymphocytes 523 [plusmn] 282, 95% receiving antiretroviral therapy (ART), duration on ART 7.1[plusmn]4.6 years) and 32 healthy controls (age 45.4[plusmn]7.2 years) without any history of coronary artery disease and with similar cardiac risk factors were prospectively recruited to undergo coronary computed tomography to assess atherosclerosis, coronary artery calcium, and plaque burden. Soluble RANKL and OPG levels were measured via ELISA.[br]Results: sRANKL was lower in HIV-infected individuals compared to controls (median 2.52 pg/ml; IQR 1.08-3.98 pg/ml vs. median 3.33 pg/ml; IQR 2.44-4.64 pg/ml, P=0.01, respectively). Amongst HIV-infected individuals, sRANKL was negatively associated with duration of HIV (Spearman [rho] -0.31, P [lt] 0.005) and positively associated with CD4 counts ([rho] 0.29, P [lt] 0.01). Exclusion of the 4 HIV-positive ART-na[iuml]ve subjects from analysis had no significant impact on the results. sRANKL was negatively associated with plaque burden ([rho] -0.41, P [lt] 0.001) and Agatston calcium score ([rho] -0.30, P[lt] 0.01) in HIV-infected individuals; however, this relationship was not observed in healthy controls. Lower OPG levels were seen in individuals with plaque compared to no plaque (median 6.29 ng/ml, IQR 3.1-18.1 ng/ml vs median 11.16 ng/ml, IQR 4.47-42.8 ng/ml; P [lt]0.05).[br]Discussion: HIV-infected individuals with lower CD4 T-lymphocytes and longer duration of disease have lower sRANKL. Furthermore, sRANKL was also lower amongst HIV-infected individuals with greater calcified coronary plaque. These data suggest that the RANKL/OPG axis may play an important role in the development of coronary artery disease in HIV patients on long standing antiretroviral therapy.[br][br]Sources of Research Support: NIH K23 HL092792 awarded to JL; Bristol Meyers Squibb awarded to SG; K24 DK064545 awarded to SG; M01 RR01066-25S1; T32 DK007028-37.[br][br]Disclosures: SG: Principal Investigator, Theratechnologies; Investigator, Theratechnologies. Nothing to Disclose: JJH, JW, SA, JL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1675 298 2079 MON-158 PO10-02 Monday 1747 2012


1744 ENDO12L_MON-159 POSTER SESSION: Vascular Biology (1:30 PM-3:30 PM) Formoterol, a Highly [beta][sub]2[/sub]-Selective Adrenergic Agonist, Induces Profound Changes in Gene Expression Governing Hypertrophy, Myogenesis, Axonogenesis and Metabolism in Human Skeletal Muscle Paul Lee, Mark J Cowley, Jing Ting Zhao, Ken KY Ho Garvan Institute of Medical Research, Sydney, Australia; Princess Alexandra Hospital, Brisbane, Australia; University of Queensland, Brisbane, Australia [beta][sub]2[/sub]-agonists are classified as performance enhancing drugs that are banned in sports. We recently reported that formoterol, a highly selective [beta][sub]2[/sub]-agonist, enhances protein anabolism in humans (1). The molecular basis of the ergogenic benefits of formoterol in humans is not well understood.[br]We have investigated the effects of formoterol on gene expression in skeletal muscle obtained from biopsies in 8 subjects before and at the end of 1 week of oral formoteral treatment at a dose of 160 ug daily. Gene expression profiling was performed using two color Agilent 44K G4112F arrays with analyses focused on muscle growth, differentiation and function. Differentially expressed genes were identified using an empirical Bayes, moderated t test, with a false discovery rate [lt] 5%. Target genes were validated by quantitative RT-PCR.[br]A total of 2641 genes were differentially expressed following formoterol treatment, of which 1312 were up-regulated and 1329 down-regulated. Formoterol increased the mRNA abundance (2-6 fold) of genes encoding signalling proteins (eg NOR-1), transcriptional (eg E2F1) and myogenic (eg GREM2) factors of pathways for protein synthesis and cell growth. It stimulated IGFI and IGFII expression and repressed those of myostatin, calpains and atrogin, factors promoting proteolysis, apoptosis and atrophy. It increased the transcription of myelin and nerve growth factors (eg PMP22 and GDNF, [gt] 4 fold) and of stretch-sensing end-plates proteins (eg ANKRD1 and CHRND, [gt]20 fold), factors enhancing axonogenesis. In contrast, formoterol modestly reduced expression of genes governing oxidative (eg PGC1a, GPI, SDHB) and fat metabolism (eg LIPE). Formoterol repressed hydroxysteroid (11[beta]) dehydrogenase 1 by 2 fold, a gene activating local cortisol production. It enhanced expression of FNDC5 (1.3 fold), recently identified as a myokine inducing brown adipose tissue differentiation (2).[br]Formoterol induced a profound effect on global gene expression in skeletal muscle. It regulates gene expression of signalling proteins, differentiation, protein synthesis, growth, apoptosis, contractile function, metabolism and local hormone synthesis. From this first human study of muscle gene expression, we conclude that formoterol stimulates skeletal muscle growth, function and regulates local hormonal milieu, changes which may enhance physical performance and muscle repair.[br][br](1)Lee P et al., Annual Scientific Meeting Endocrine Society 2011 Abstract P1-472. (2)Bostr[ouml]m P et al., Nature. 2012; 481: 463-8.[br][br]Sources of Research Support: Paul Lee is supported by a postgraduate scholarship from NHMRC, Australia.[br][br]Nothing to Disclose: PL, MJC, JTZ, KKYH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 910 298 2080 MON-159 PO10-02 Monday 1748 2012


1745 ENDO12L_MON-160 POSTER SESSION: Vascular Biology (1:30 PM-3:30 PM) Comparative Proteomic Study of Cardiomyocyte Lipid Droplets Reveals That the Location of ATGL Is Involved in Heart Failure Huina Zhang, Linghai Li, Pingsheng Liu Institute of Biophysics, Beijing, China Excessive storage of lipid droplet (LD) within cardiac muscle due to aberration of lipid metabolism and lipid traffic has been linked to the development and progression of several common cardiomyopathies including ischemia-reperfusion injury, cardiac arrhythmia, and heart failure. The neutral lipid retention in intracellular LDs is accurately regulated by proteins on the lipid droplets. Therefore, identification and comparison of the myocardial LD proteome between normal and overload-induced failing hearts may provide useful insights to dissect the molecular mechanism of metabolic disorder in heart diseases. In the present work, we established a method to purify LDs from rat heart and identified 194 LD-associated proteins in normal rat heart through mass spectrometry analysis. Besides the well studied LD protein members including Perilipin family proteins, membrane traffic proteins and enzymes of lipid metabolism, our proteomic research also found 77 newly identified proteins on heart LDs, including muscle-specific resealing protein-dysferlin. Immunoblotting and immunoflurorescence were used to further verify the expression and location of dysferlin and its-associated proteins VCP, annexin 2, and caveolin 3. Moreover, using truncated dysferlin, we demonstrated that the CH2 domain of dysferlin was responsible for its LD targeting.[br]The comparative proteomic results exhibited the dynamic changes in LD-associated proteins in normal and failing heart. 12 proteins including Perilipin family proteins and triglyceride metabolic proteins were identified using mass spectrometry and some of them were validated by immunoblotting. Among these proteins, the level of ATGL on failing heart LD was dramatically decreased compare to normal heart while no change of the protein expression was detected. Further study indicated that OXPAT was involved in the different location of ATGL. In addition, overexpression of OXPAT and ATGL in heart by adenovirus ameliorated the cardic function under heart failure.[br]Our results suggest that LDs may be involved in membrane integrity of cardiomyocytes through dysferlin mediation, and increasing the location of ATGL on LD may be a possible treatment for failing heart.[br][br]Sources of Research Support: The Ministry of Science and Technology of China (Grant No. 2009CB919000; Grant No. 2010CB833703), the National Natural Science Foundation of China (Grant No. 30871229 and Grant No. 30971431).[br][br]Nothing to Disclose: HZ, LL, PL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 402 298 2081 MON-160 PO10-02 Monday 1749 2012


1746 ENDO12L_MON-161 POSTER SESSION: Vascular Biology (1:30 PM-3:30 PM) Transcriptional Regulation of Endothelin-1 Expression by Advanced Glycation End-Products in Human Aortic Endothelium Is Mediated Via NF-[kappa]B and AP-1 Christos Adamopoulos, Christina Piperi, Georgia Dalagiorgou, Antonios Nikolaou Gargalionis, Eleni Artemiou Kandaraki, Evanthia Diamanti-Kandarakis, Athanasios Georgiou Papavassiliou Medical School, University of Athens, Athens, Greece; Medical School, University of Athens, Athens, Greece [bold]Introduction:[/bold] Advanced Glycation End-products (AGEs) are produced by the non-enzymatic glycation of proteins, lipids and nucleic acids, resulting in an overload of highly reactive molecules of endogenous or exogenous (dietary) origin. Increased AGE levels in circulation and concomitant elevated tissue deposition have been associated with diabetic complications, atheromatosis, ageing and more recently with polycystic ovary syndrome pathogenesis. Interaction of AGEs with their receptor RAGE (Receptor for AGEs) activates intracellular signaling pathways which induce targeted gene expression in endothelium including upregulation of cell adhesion molecules and endothelin-1 (ET-1), implicated in vascular injury and endothelial dysfunction. [bold]Aim:[/bold] To explore the molecular mechanism of AGE-induced regulation of [italic]ET-1[/italic] gene/protein expression in human endothelial cells and investigate its functional relevance in normal rat vascular endothelium. [bold]Methods [amp] Materials:[/bold] Human aortic endothelial cells (HAECs) were treated with increasing concentrations (100, 200 ug/ml) of AGE-bovine serum albumin (AGE-BSA) in different time-points (24, 48, 72 h). The inducible expression of ET-1 and RAGE was studied by quantitative real-time polymerase chain reaction (real-time PCR) and flow cytometry. Transcription factor NF-kB (Nuclear Factor-kappaB) and AP-1 (Activator Protein-1) binding capacity was investigated by electrophoretic-mobility shift assay (EMSA). Functional relevance of AGE signaling in ET-1 regulation was investigated immunohistochemically in normal aortic endothelium of rats fed with low- or high-AGE content diet. [bold]Results:[/bold] Treatment of HAECs with AGE-BSA induced ET-1 transcription and protein expression in a time- and dose- dependent manner. This induction was mediated by activation and binding to [italic]ET-1[/italic] gene promoter of NF-kB and AP-1 transcription factors. RAGE expression in HAECs remained unchanged after AGE treatment. In addition, increased expression of AGEs, RAGE and ET-1 was observed in the aortic endothelium of normal rats fed with high-AGE diet compared with controls. [bold]Conclusion:[/bold] AGE-RAGE signaling induces ET-1 protein expression in endothelium through regulation of [italic]ET-1[/italic] gene promoter by the transcription factors, NF-kB and AP-1 constituting a molecular mechanism that potentially contributes to the characteristic endothelial dysfunction of obesity, diabetic microvascular complications, atherogenesis and polycystic ovary syndrome.[br][br]Sources of Research Support: This research has been co-financed by European Union (European Social Fund - ESF) and Greek national funds through the Operational Program [quot]Education and Lifelong Learning[quot] of the National Strategic Reference Framework (NSRF) - Research Funding Program: Heracleitus II. Investing in knowledge society through the European Social Fund.[br][br]Nothing to Disclose: CA, CP, GD, ANG, EAK, ED-K, AGP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 610 298 2082 MON-161 PO10-02 Monday 1750 2012


1747 ENDO12L_MON-162 POSTER SESSION: Vascular Biology (1:30 PM-3:30 PM) Influence of Renin-Angiotensin System on Myocardial Remodeling in Experimental Hyperthyroid Cardiomyopathy Yang-Seon Yi, Min Jung Bae, Won Jin Kim, Kyoung Im Cho, Seong Man Kim, Yun Kyung Jeon, Sang Soo Kim, Bo Hyun Kim, Yong Ki Kim, In Joo Kim Pusan National University School of Medicine, Busan, Korea; Maryknoll Medical Center, Busan, Korea; Pusan National University School of Medicine, Busan, Korea; Kim Yong Ki Internal Medicine Clinic, Busan, Korea [bold]BACKGROUND:[/bold] This study aimed to evaluate the effects of irbesartan, a renin-angiotensin system (RAS) blocking agent and dronedarone, which blocks multiple cardiac ion channels and [beta]-adrenoceptors, on the functional and structural manifestations of experimental hyperthyroid cardiomyopathy.[br][bold]METHODS:[/bold] A rat model of hyperthyroidism was established by daily intraperitoneal injections of L-thyroxine (T[sub]4[/sub], 100 mg/kg per day) for 28 days. Forty Sprague-Dawley rats were randomly divided into four groups (n = 10 each): control group, T[sub]4[/sub] group (T[sub]4[/sub] alone), T[sub]4 [/sub]plus irbesartan group (T[sub]4[/sub]-Irb, 30 mg/kg irbesartan), and T[sub]4[/sub] plus dronedarone group (T[sub]4[/sub]-Dro, 100 mg/kg dronedarone). All rats underwent echocardiography and cardiac chamber size and strain parameters were measured after 28 days of treatments. Cardiomyocyte diameter and cardiac fibrosis were measured using pathological analysis.[br][bold]RESULTS:[/bold] Heart rate was increased in T[sub]4[/sub] alone and T[sub]4[/sub]-Irb groups. When compared with control, T[sub]4[/sub] alone showed significantly reduced ejection fraction (EF; 72.1 [plusmn] 5.49 vs. 77.6 [plusmn] 5.01 %, P [lt]0.05) and LV longitudinal strain (LV strain; -16.0 [plusmn] 6.27 vs. -22.7 [plusmn] 5.19 %, P[lt]0.05). However, compared with control, T[sub]4[/sub]-Irb showed similar EF (77.6 [plusmn] 3.85 %.) and LV strain (-21.4 [plusmn] 1.84 %). Morphologically, T[sub]4[/sub] alone group showed significantly increased cardiomyocyte width (25.3 [plusmn]1.89 vs. 18.9[plusmn] 1.14 [mu]m in control, P [lt] 0.001) and fibrosis, which is prevented by irbesartan treatment (cardiomyocyte width; 18.90 [plusmn] 1.14 vs. 25.3 [plusmn]1.89 [mu]m in T[sub]4[/sub] alone, P [lt] 0.05). Although T[sub]4[/sub]-Dro showed no significant reduction in cardiomyocyte width and fibrosis compared with T[sub]4[/sub] alone, EF and LV strain was improved compared with T[sub]4[/sub] alone.[br][bold]CONCLUSIONS:[/bold] Irbesartan could significantly attenuate structural and functional remodeling induced by experimental hyperthyroid cardiomyopathy, which is not found with dronedarone with regulation of heart rate. These results may implicate the potent role of RAS in hyperthyroid cardiomyopathy beyond tachycardia.[br][br]Nothing to Disclose: Y-SY, MJB, WJK, KIC, SMK, YKJ, SSK, BHK, YKK, IJK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 78 298 2083 MON-162 PO10-02 Monday 1751 2012


1748 ENDO12L_MON-163 POSTER SESSION: Vascular Biology (1:30 PM-3:30 PM) Salvianolic Acid B Inhibits Proliferation in Vascular Smooth Muscle Cells through Regulating ROS Via Heme Oxygenase-1 Induction Hyun Jung Lee, Eun Jig Lee Yonsei University College of Medicine, Seoul, Korea; Yonsei University College of Medicine, Seoul, Korea; Northwestern University Feinberg School of Medicine, Chicago, IL The reactive oxygen species (ROS)-induced vascular smooth muscle cell (VSMCs) proliferation is involved in the development of atherosclerosis and restenosis. Also, ROS is triggered by multiple factors including cytokines such as growth factors such as platelet-derived growth factor (PDGF), which are produced by platelets, VSMCs, and ECs in the injured vascular wall. This study was designed to investigate the effects of salvianolic acid B (Sal B), one of the compounds in Danshen on VSMCs. Stimulating VSMCs with PDGF-BB to induce proliferation, migration and ROS generation was used to identify the roles of Sal B. Sal B treatment significantly inhibited PDGF-BB-induced cell proliferation and migration, and reduced intracellular ROS generation by PDGF-BB in a dose-dependent manner in VSMCs. Sal B significantly induced heme oxygenase-1 (HO-1) protein expression and inhibited PDGF-BB-induced phosphorylation of AKT by scavenging ROS. Furthermore, Sal B activated the translocation of the transcription factor nuclear factor-E2-related factor 2 (Nrf2) in VSMCs. Our data demonstrated that Sal B inhibits PDGF-BB-induced cell proliferation and migration in VSMCs by increasing the expression of HO-1 via Nrf2 activation and ROS scavenging. This suggests Sal B may be a promising agent to prevent atherosclerosis and restenosis.[br][br]Nothing to Disclose: HJL, EJL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 676 298 2084 MON-163 PO10-02 Monday 1752 2012


1749 ENDO12L_MON-164 POSTER SESSION: Vascular Biology (1:30 PM-3:30 PM) Inhibition of Endoplasmic Reticulum Stress by Vitamin D in Endothelial Cells Anna Szafran-Swiertlik, Syed H Jafri, Jessie Wang, Sophie Wang, Arshag D Mooradian, Michael J Haas University of Florida, Jacksonville, FL Damage to endothelial cells is regarded to play an important role in atherosclerosis. Pro-inflammatory cytokines, fatty acids, and hyperglycemia have been shown to disrupt endothelial cell barrier function as well as enhance the expression of chemoattractant molecules by inducing cellular stress. To determine whether or not vitamin D inhibits endoplasmic reticulum (ER) stress, human umbilical vein endothelial cells were treated with tunicamycin with or without vitamin D and several vitamin D-related analogs. When added together, vitamin D prevented ER stress. However, the inhibitory effect was much stronger when cells were pre-treated with vitamin D for 24-hours. Endoplasmic reticulum stress was not inhibited by 25-OH vitamin D[sub]3[/sub] or the non-calcemic vitamin D analog EB1089. However both ZK191784 and the vitamin D metabolite 24,25-dihydroxyvitamin D[sub]3[/sub] were as effective as vitamin D in preventing ER stress. Similar effects were observed when ER stress was induced by hyperglycemia. These results suggest that vitamin D has a protective effect on vascular endothelial cells.[br][br]Nothing to Disclose: AS-S, SHJ, JW, SW, ADM, MJH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1681 298 2085 MON-164 PO10-02 Monday 1753 2012


1750 ENDO12L_MON-165 POSTER SESSION: Vascular Biology (1:30 PM-3:30 PM) Elevated Fibroblast Growth Factor-21 Is Associated with Increased Carotid Intima-Media Thickness Wing Sun Chow, Michele Mae Ann Yuen, Aimin Xu, Carol Ho Yi Fong, Cheng Chen, Stephen Chi Wai Cheung, Ming Tak Chau, Nelson Ming Sun Wat, Rachel Lai Ching Wong, Karen Siu Ling Lam The University of Hong Kong, Hong Kong, Hong Kong; The University of Hong Kong, Hong Kong, Hong Kong; Queen Mary Hospital, Hong Kong, Hong Kong Objective: Elevated circulating concentrations of fibroblast growth factor-21 (FGF21), a recently recognised regulator of glucose and lipid metabolism, have been reported in type 2 diabetes, coronary artery disease, and in dialysis-dependent patients with end-stage renal failure. In this study, we investigated the relationship of serum FGF21 levels with carotid atherosclerosis in subjects with normal serum creatinine level.[br]Design and Methods: In 670 Chinese subjects, anthropometric and biochemical parameters, serum FGF21 concentrations and carotid intima media thickness (IMT) were measured. The relationships between serum FGF21 concentrations, various cardiometabolic risk factors, and IMT were analysed.[br]Results: Carotid IMT was measured in 285 men and 385 women mean age 58.2[plusmn]12.9 years). Men had higher IMT (0.73[plusmn]0.17 mm vs. 0.68[plusmn]0.14 mm, p[lt]0.001). Serum FGF21 was similar between men and women, being 248[159.6-368.2] ng/l and 251 [136.8-415.2] ng/l, median [Inter-quartile range] respectively, and correlated positively with age (p[lt]0.001), anthropometric parameters (p[lt]0.05), blood pressure, LDL, triglycerides, serum creatinine level, and negatively with HDL and estimated glomerular filtration rate (eGFR) (all p[lt]0.001). A significant positive correlation was found between serum FGF21 concentration and carotid IMT (r=0.32; p[lt]0.001) in women but not in men. This relationship remained significant even after adjusting for age, waist circumference, hypertension, fasting glucose, serum creatinine level, dyslipidaemia and smoking status (standardized beta=0.103; p=0.021). Using eGFR instead of serum creatinine level in the regression model did not alter the finding.[br]Conclusions: Our results suggested a positive correlation between circulating levels of FGF21 and carotid IMT, a marker of carotid atherosclerosis, being demonstrable in women. This relationship was independent of various cardiometabolic risk factors. These observations would support a role for FGF21 or FGF21 resistance in the pathogenesis of atherosclerosis.[br][br]Sources of Research Support: Collaborative Research Grant (HKU3/CRF/09) from the Hong Kong Research Grants Council.[br][br]Nothing to Disclose: WSC, MMAY, AX, CHYF, CC, SCWC, MTC, NMSW, RLCW, KSLL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 740 298 2086 MON-165 PO10-02 Monday 1754 2012


1751 ENDO12L_MON-166 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Two Mutations Causing Monogenic Diabetes Identified during Short Stature Investigation in Brazilian Families Milena G Teles, Ericka B Trarbach, Alexsandra C Malaquias, Alexander A Jorge Faculdade de Medicina da Universidade de S[atilde]o Paulo (FMUSP), Sao Paulo, Brazil The prevalence of monogenic diabetes (MD) varies among different populations. Since there are differences in prognosis and treatment of MD, a precise diagnosis is mandatory. To date, few mutations in GCK were described in Brazilian patients. Here we report two cases that were referred to our hospital for investigation of short stature and were diagnosed with monogenic diabetes. Case 1: a prepubertal 14-year-old boy with height of 130 cm (-3.5 SD), weight of 25.5 kg was evaluated for constitutional delay of growth and puberty. During follow-up, he started puberty at the age of 16 and his adult height was 161 cm. Laboratory evaluation showed abnormal fasting glucose [105-131 mg/dL (5.8 [ndash] 7.3 mmol/L)] and HbA1c levels (6.6 %). Serum anti-GAD, anti-IA2, anti-ICA were negative. Oral glucose tolerance test showed an increase of 55 mg/dL in glucose levels after 2 h. His father also had slightly elevated blood glucose levels as well as the paternal grandmother. A heterozygous missense mutation in exon 6 of GCK, c.571C[gt]T (p.R191W) was identified. The father and the grandmother also harbor the mutation, but not the mother who does not have hyperglycemia. In total, 19 family members were screened for p.R191W mutation and 11 had the mutation in heterozygous state. All affected patients had mild elevated glycemia. Case 2 is a 4-year-old boy with 95 cm (-1.5 SD) and 15.8 kg. During follow-up (3 years) he remained in the fifth percentile in the growth chart. Laboratorial tests revealed no alterations other than elevated glycemia [118 mg/dL (6.6 mmol/L)]. [beta]-cell antibodies were negative. His father, a paternal aunt and the paternal grandmother had also slightly elevated glycemia, whereas his mother had normal glycemia. GCK sequencing identified a heterozygous missense mutation in exon 5, c.661G[gt]A (p.E221K) in index patient as well in all family members with hyperglycemia. Even though already described in other ethnicities, the GCK mutations p.R191W and p.E221K were never identified in Brazilian families. The identification of GCK mutations in patients with hyperglycemia has implications for their clinical course and management. Hence, GCK mutation screening should be considered in patients with chronic asymptomatic hyperglycemia, with early age of onset, negative [beta] -cell antibodies as we observed in these patients investigated for short stature.[br][br]Nothing to Disclose: MGT, EBT, ACM, AAJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1330 299 2087 MON-166 PO47-01 Monday 1755 2012


1752 ENDO12L_MON-167 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Metreleptin Treatment in Acquired Generalized Lipodystrophy: Consistent Metabolic Effects in Three Patients Presenting with Distinct Autoimmune Conditions Nevin Ajluni, Adam Neidert, Carmen Tapiador, Barbara S Adams, Jean L Chan, Elif A Oral University of Michigan, Ann Arbor, MI; East Tennessee Children[apos]s Hospital, Knoxville, TN; Amylin Pharmaceuticals, Inc, San Diego, CA Background: Acquired generalized lipodystrophy (AGL) is a rare heterogeneous disorder characterized by fat loss, associated with severe insulin resistance, diabetes, dyslipidemia, and leptin deficiency. While metreleptin therapy has been shown to improve metabolic abnormalities in lipodystrophy patients, the effect in AGL patients with active autoimmune diseases warrants further characterization. Between 3/2009 and 10/2011, 3 pediatric patients with AGL and unusual presentations received metreleptin, an experimental medication, in an ongoing open-label expanded access program (sponsored by Amylin Pharmaceuticals, Inc.) at University of Michigan.[br]Clinical Cases: AGL-1 (9 y, female) presented with severe hypertriglyceridemia, uncontrolled diabetes and hyperphagia associated with active juvenile dermatomyositis. After 15 months of metreleptin, triglycerides decreased from 10,623 mg/dL to a low of 123 mg/dL (n[lt]150 mg/dl), and HbA1c decreased from 9.1% to a low of 7.2% (n[lt]6.3%) while daily insulin dose was reduced from 400 to 60 units. Functional capacity improved and muscle inflammation (evaluated by extremity MRI) was no longer present at 2 years while metabolic benefits were sustained.[br]AGL-2 (9 y, male) presented with autoimmune hepatitis and AGL. He did not have diabetes but had hypertriglyceridemia (354 mg/dL), hyperphagia leading to aggressive night-time food-seeking behavior, and markedly abnormal aminotransferases (ALT 419 U/L, n[lt]35 U/L; AST 208 U/L, n[lt]30 U/L). After 6 months of metreleptin, triglycerides decreased to 118 mg/dL and ALT/AST to 89/60 U/L. Importantly, food intake normalized and aggressive behavior resolved.[br]AGL-3 (16 y, female) presented with difficult to control diabetes and insulin resistance. She had a diagnosis of Graves[apos] disease prior to development of AGL. After 3 months of metreleptin, HbA1c decreased from 10.2% to 5.0%, triglycerides decreased from 1,845 to 324 mg/dL and aminotransferases decreased (ALT 259 to 40 U/L) with sustained effects at 9 months. Despite the detection of anti-GAD autoantibody at baseline, she was able to discontinue insulin ([gt]300 units/day) as well as pioglitazone.[br]Clinical Lessons: These cases demonstrated that the presentation of AGL can be quite heterogeneous and associated with diverse autoimmune diseases. Metreleptin therapy resulted in substantial improvements in metabolic abnormalities, liver function tests, and/or hyperphagia without evidence of worsening of the associated autoimmune disease.[br][br]Disclosures: JLC: Employee, Amylin Pharmaceuticals. EAO: Principal Investigator, Amylin Pharmaceuticals. Nothing to Disclose: NA, AN, CT, BSA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1096 299 2088 MON-167 PO47-01 Monday 1756 2012


1753 ENDO12L_MON-168 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Recurrent Diabetic Ketoacidosis in a Patient with Type B Insulin Resistance Syndrome Due to Suboptimal Insulin Therapy Manan Shah, Muhammad Mahmood, Mohammad Ansari, Vinaya Simha Texas Technical University Health Sciences Center, Odessa, TX [bold]Background: [/bold]Syndromes of extreme insulin resistance can result from either insulin receptor mutations (Type A) or circulating antibodies to insulin receptor (Type B). They are characterized by severe hyperglycemia and high insulin requirements, but unprovoked diabetic ketoacidosis (DKA) is not commonly reported. We report a patient with type B insulin resistance syndrome (IR) who developed recurrent severe unprovoked DKA even on insulin therapy.[br][bold]Clinical case[/bold]: A 30 year old African American lady with recent diagnosis of type 2 DM on Metformin therapy (HbA1c of 6.2%), presented to the hospital with mild DKA in association with urosepsis, which resolved with intravenous (iv) insulin and antibiotics. She was discharged on a basal/bolus regimen of glargine and aspart (about 50 units/day) after achieving satisfactory glucose control. She was readmitted after a month with DKA and HbA1c of 7.9%. She required considerably higher insulin doses to reverse ketosis, and was finally discharged after a week on 50 units of glargine twice a day and 30 units of aspart with each meal. However, she returned to the hospital in 2 weeks with severe DKA and hyperglycemia despite insulin therapy which had been increased to 600 units/day. An extensive workup for precipitating infections including pan-culture and lumbar puncture were negative, and she required iv insulin infusion rate of 75-100 units/h for over 5 days to reverse the metabolic decompensation. She was eventually started on Regular insulin U500 1cc four times a day (2000 units/day), and has remained free of ketoacidosis for the last 6 months. Serum C-peptide and insulin (both total and free) levels were markedly elevated during hospitalization (9.3 ng/ml, 1371 uU/mL and 865uU/mL, respectively). She was of lean build and had acanthosis over the neck and face raising suspicion of Type B IR, which was confirmed by detection of insulin receptor antibodies.[br][bold]Clinical lesson:[/bold] Unprovoked DKA can be a feature of Type B IR when insulin therapy is suboptimal. It has been shown previously that these subjects are very resistant to oxidative glucose disposal, and an insulin infusion rate of 1000mU/kg.min was necessary to stimulate glucose oxidation and suppress lipid oxidation. Seemingly high insulin therapy (500-1000 units/day) may not be effective in suppressing lipolysis and ketogenesis in these rare patients leading to unprovoked DKA.[br][br]Nothing to Disclose: MS, MM, MA, VS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1642 299 2089 MON-168 PO47-01 Monday 1757 2012


1754 ENDO12L_MON-169 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) A Diagnostic Dilemma of Hypoglycemia in End-Stage Liver Disease: Is It an Insulinoma? Abdulrahman Alkabbani, Revital Gorodeski-Baskin, Marwan Hamaty Endocrinology and Metabolism, Cleveland, OH [bold]Introduction[/bold]: Liver cirrhosis is associated with peripheral hyperinsulinemia due to decreased clearance of insulin. Spontaneous hypoglycemia in the setting of end-stage liver disease (ESLD) represents a diagnostic challenge due to co-existing hyperinsulinemia.[br][bold]Clinical case: [/bold]A 34-year old woman with ESLD secondary to Wilson[apos]s disease was evaluated for liver transplantation (Tx). She complained of frequent episodes of near fainting, usually a few hours after a meal. When symptomatic, she had a markedly elevated proinsulin of 220 pmol/L (3-20), mildly elevated C-Peptide 4.2 ng/mL (0.8-3.2), low plasma glucose (GLU) 46 mg/dL (65-100), and negative serum sulfonylurea screen [amp] insulin antibodies. ACTH stimulation test resulted in peak cortisol of 16.3 ug/dL (n= [gt]18) in the setting of a low albumin level 1.8 g/dL (3.5-5) and low pre-albumin 7 mg/dL (18-45). Creatinine was normal at 0.6 mg/dL (0.7-1.4), and she had a mild primary hypothyroidism with positive thyroid peroxidase antibodies. Liver transplantation was postponed despite the availability of a donor because the patient was deemed not a good candidate for liver Tx due to the questionable insulinoma under investigation.[br]The 72-hour fast test resulted in hypoglycemia (GLU [lt]55 mg/dL) at the 10[sup]th[/sup] hr and was aborted at the 15[sup]th[/sup] hr. Results at those 2 time points were as follows: GLU 53, 38 mg/dL; Insulin 9.1, 13.4 uU/mL (1-24); proinsulin 170, 130 pmol/L; c-peptide 2.7, 2.4 ng/mL; betahydroxybutyrate 0.02, 0.02 mmol/L, respectively. Insulin antibody was [lt] 5%, and serum sulfonylurea screen was negative. Pancreatic CT was negative. The cause of hypoglycemia was determined to be caused by ESLD based on the finding of significant elevation of insulin and proinsulin in association with only mild elevation of C-peptide, the latter not greatly affected by liver failure. Seventeen days later, she received an orthotopic liver Tx and required insulin therapy for a short period of time due to steroid use. She was not hypoglycemic on follow up visits off steroids, and maintained normal glucose values.[br][bold]Conclusion:[/bold] Hyperinsulinemia is a common manifestation of ESLD. Reduced hepatic gluconeogenesis, and reduced insulin degradation in ESLD causing hypoglycemia may represent a clinical and biochemical picture similar to insulinoma. C-peptide levels serve as an important differentiating factor, which were significantly lower than proinsulin and insulin in our patient. Expectedly, liver Tx resulted in resolution of hypoglycemia.[br][br]Nothing to Disclose: AA, RG-B, MH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2135 299 2090 MON-169 PO47-01 Monday 1758 2012


1755 ENDO12L_MON-170 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Pseudohypoglycemia with Hospital-Based Assisted Monitoring of Blood Glucose Gillian Boyd-Woschinko, Ronald Tamler The Mount Sinai Hospital, New York, NY [underline]Background:[/underline] Hypoglycemia in hospitalized patients is common and has serious implications. Clinicians base glycemia management on assisted monitoring of blood glucose (AMBG). We present a case that illustrates the pitfalls, differential diagnosis and management of pseudohypoglycemia with AMBG.[br][underline]Case:[/underline] A 34 year old woman with history of end-stage renal disease, HIV and recent myocardial infarction presented to the hospital with hypovolemic shock after dialysis. Her hemoglobin (Hb) was 3 g/dL (normal 11.7-15.0 g/dL), and she was admitted to the ICU. The patient received broad spectrum antibiotics, stress-dose glucocorticoids and multiple blood transfusions, and her Hb improved to 9 g/dL. No infection was found; antibiotics and steroids were stopped after a seven day course. The patient had no history of diabetes mellitus and was consuming a renal diet. Her whole blood glucose was 73 mg/dL (normal 60-120 mg/dL) by GEM [reg] analyzer in the ICU, but AMBG was 11mg/dL and [lt]10 mg/dL after transfer to the general wards. She experienced no diaphoresis, anxiety, or neurocognitive deficits. The patient was urgently given 50ml of IV Dextrose 50% and started on IV dextrose 10% infusion, with no change in AMBG or clinical status. ACTH-stimulation test was within normal limits. The consulted endocrine team noted on physical exam normal nutritional status, cyanotic and cold digits bilaterally, and an arterial-venous graft on the left forearm with a thrill. The team suspected pseudohypoglycemia due to impaired peripheral microcirculation and ordered an AMBG site change from the patient[apos]s fingers to her earlobes. Subsequent results were consistently 70-90 mg/dL without IV dextrose infusion.[br][underline]Conclusion[/underline]: While this particular vignette showcases a patient with vascular disease and recent hypovolemic shock that had unreliable AMBG from the finger tips, it enables a general overview of risk factors for pseudohypoglycemia and ways to achieve reliable glucose measurements. Clinicians must be mindful of this condition in order to avoid unnecessary tests and treatments.[br][br]Nothing to Disclose: GB-W, RT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1558 299 2091 MON-170 PO47-01 Monday 1759 2012


1756 ENDO12L_MON-171 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) Celexa Induced Hypoglycemia Mansi Mehta, Kenneth H Hupart NuHealth, Nassau University Medical Center, East Meadow, NY; St John[apos]s University, Queens, NY; Albert Einstein College of Medicine, Bronx, NY 56-year-old African-American male from nursing home was admitted to the hospital with an acute drop in his hemoglobin concentration to 5.6 mg/dL (baseline Hgb of 10 mg/dL). His past medical history was notable for type 2 diabetes controlled with diet, end-stage renal disease on hemodialysis (HD), hepatitis B and C, HIV (last viral load undetectable), hypertension and depression. His nursing home medications included aspirin, hydralazine, lisinopril, lamivudine, zidovudine, raltegravir and sertraline. Patient received packed red cell transfusion and an upper endoscopy/colonoscopy showed clean shallow ulcer at ileocecal valve. During his hospital course, patient became increasingly somnolent and sertraline was switched to citalopram 20 mg oral daily. A week later, he started complaining of sweating, tremors, anxiety and agitation with random glucose values ranging from 37 to 56 mg/dL (both before and after meals) that were confirmed with venous blood samples. Hypoglycemia was relieved with meals. Common causes of hypoglycemia were ruled out by thyroid and liver related tests, anti-insulin antibodies, adrenal insufficiency, insulinoma and surreptious abuse of sulfonylureas or insulin. His 72-hour fast was aborted, as he had low glucose of 45 at the end of 36 hours. Patient was experiencing new onset hypoglycemia and the only change made was addition of citalopram. Hence, because of rare reports of citalopram induced hypoglycemia this medication was discontinued, and within 24 hours patient[apos]s daily hypoglycemia episodes ceased.[br]Citalopram (Celexa) is a selective serotonin reuptake inhibitor (SSRI) commonly used in the treatment of depression in diabetics. Within the class of SSRI[apos]s hypoglycemia is rarely reported. We found 2 case reports of citalopram causing hypoglycemia and 2 cases reported of overdose. We were impressed by the fact that hypoglycemia occurred very soon after starting citalopram, and resolved very rapidly. However, the exact mechanism is still unclear. In a hospital setting, in a patient with multiple co-morbities it is difficult to pinpoint the exact etiology of hypoglycemia. And we should be aware about this important side effect of the drug as it is easily managed. Hence, our goal is to call attention to this clinical problem.[br][br]Nothing to Disclose: MM, KHH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2320 299 2092 MON-171 PO47-01 Monday 1760 2012


1757 ENDO12L_MON-172 POSTER SESSION: Diabetes [amp] Insulin Resistance Case Reports: Unusual Diagnostic Considerations [amp] Rare Birds (1:30 PM-3:30 PM) A Case of Artifactual Hypoglycemia in a Patient with Hyperleukocytosis Secondary to Alcoholic Hepatitis Pooja Raghavan, Dhyan Rajan, Christopher Tan, Chinedu Nweke, Salini C Kumar Mount Carmel Medical Center, Columbus, OH; Nassau University Medical Center, East Meadow, NY [bold]Background[/bold][br]Artifactual hypoglycemia (AH) is the presence of a falsely decreased laboratory measurement of blood glucose. Increased in vitro glycolysis by leukocytes has been well documented; however this occurs primarily in the setting of myeloproliferative disorders, leukemoid reactions, and leukemias. We present a case of AH in a patient with alcoholic hepatitis and hyperleukocytosis.[br][bold]Case[/bold][br]A 27-year old male with a history of alcohol abuse presented to the emergency department with severe abdominal pain. Physical examination was remarkable for right upper quadrant tenderness. Laboratory evaluation revealed marked leukocytosis of 46 K/mm3 with neutrophilia (84%); liver tests were consistent with alcoholic hepatitis. Throughout the course of the patient[apos]s hospital stay, serum blood glucose levels ranged from 24-69 mg/dl; while concurrent capillary blood sampling ranged from 90-120 mg/dl. The patient had no evidence of neuroglyopenic or adrenergic symptoms of hypoglycemia. He maintained a good appetite, and did not have diarrhea or vomiting. The patient was not on medications that cause hypoglycemia. His serum TSH and cortisol levels were normal. His leukocyte count remained elevated ranging from 32-50 K/mm3. A diagnostic evaluation for infection was negative. A bone marrow biopsy performed was negative for both myelodysplasic and myeloproliferative syndromes. Liver biopsy confirmed the presence of alcoholic hepatitis. He was treated conservatively; with a concomitant decrease in white blood cell counts. As the patient[apos]s leukocyte count normalized so did his serum glucose levels. It was later confirmed that blood samples were not collected in tubes containing antiglycolytic agents; and often up to two hours would elapse from phlebotomy to sample analysis.[br][bold]Conclusion[/bold][br]Artifactual hypoglycemia (AH) in patients with hyperleukocytosis secondary to alcoholic hepatitis is rare. Although severe liver disease may result in an interruption of gluconeogenesis and cause true hypoglycemia, AH should be considered in asymptomatic patients with hyperleukocytosis from alcoholic hepatitis. This artifact can be prevented by the utilization of glycolysis inhibitors and prompt delivery of the collected specimen to the laboratory. Clinicians should be cognizant of AH, as this awareness may prevent expensive and unnecessary diagnostic evaluations.[sup][1][/sup][br][br](1) Ybarra J, Isern J. Leukocytosis-Induced Artifactual Hypoglycemia. Endocrine Journal. 2003. 50 (4); 481-482.[br][br]Nothing to Disclose: PR, DR, CT, CN, SCK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1245 299 2093 MON-172 PO47-01 Monday 1761 2012


1776 ENDO12L_MON-191 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) A Retrospective Study on Metformin Efficacy in Neuropathic Pain Magdalena Szkudlinska, Amber Taylor, Prathima Guruguri, Emil Annabi, Theodore Price, Hussein Yassine University of Arizona, Tucson, AZ The oral hypoglycemic agent metformin is a long approved and widely used treatment for type 2 diabetes. While its mechanism has never been fully elucidated, recent research has found that metformin exhibits anti-oxidative properties and activates 5[apos] adenosine monophosphate-activated protein kinase (AMPK). We have recently shown that metformin and other AMPK activators reverse neuropathic pain in preclinical studies using rats and mice. Therefore, we hypothesized that metformin might decrease lumbar radiculopathy pain in humans.[br]Methods: We performed a chart review on over 2000 patients who sought care from a university pain specialist for lumbar radiculopathy between Jan 2008-Nov 2011 and were asked to fill out a standardized questionnaire regarding their pain qualities. Patients coincidentally taking metformin at the time of the visit were designated the treatment group and were matched with subjects having lumbar radiculopathy but not on metformin therapy. Patients with diabetic complications including pre-existing peripheral neuropathy were excluded. We matched the treatment and control groups on age, sex, BMI, socioeconomic status, and comorbidities; stroke, depression, diabetes, heart disease, and pain medications (in particular opioids, TCAs, and benzodiazepines). The outcomes were individual qualitative and quantitative pain scores, total pain experience, pain at the time of visit, and interference with daily functions. Responses on pain questionnaires were analyzed using Pearson chi-square and t-tests.[br]Results: We selected a total of 45 patients on metformin (age 59.3, BMI 33.2) and 58 controls (age 55.1, BMI 31.3), most of whom had diabetes. Mean number of pain medications taken were 2.5 in treatment group vs. 2.7 in controls (p=0.56), with mean onset of pain 9.9 years ago on metformin and 10.9 years ago for those not on therapy (p=0.64). Individual subtypes of pain were analyzed; both [ldquo]hot burning[rdquo] and [ldquo]aching[rdquo] were more likely to be observed in those not taking metformin vs. the treatment group (p=0.08 and 0.02 respectively). Those on metformin were less likely to experience interference with their working routine (p=0.10), interference with relations with others (p=0.09), and decreased enjoyment of life (p=0.02).[br]Conclusion: Metformin therapy is associated with decreased severity of neuropathic pain independent of diabetes status. This finding supports preclinical studies and warrant a prospective study of metformin on neuropathic pain.[br][br]Nothing to Disclose: MS, AT, PG, EA, TP, HY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1184 300 2112 MON-191 PO11-01 Monday 1780 2012


1777 ENDO12L_MON-192 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Saxagliptin vs Uptitrated Metformin Extended Release: Effect of Baseline HbA[sub]1c[/sub] on Efficacy and Safety Boaz Hirshberg, Vivian Fonseca, Gianmaria Minervini, Mark Donovan, Peter Ohman AstraZeneca, Wilmington, DE; Tulane University Health Sciences Center, New Orleans, LA; Bristol-Myers Squibb, Princeton, NJ In a phase IIIb trial of patients with type 2 diabetes and inadequate glycemic control on metformin extended release (MET XR) 1500 mg/d, adding saxagliptin (SAXA) 5 mg/d (SAXA + MET XR) for 18 weeks produced statistically significantly greater improvement in glycemic control than uptitrating MET XR to 2000 mg/d in the overall study ([italic]P[/italic][lt]0.0001) and was well tolerated (NCT00960076). Because efficacy and tolerability may vary with degree of glycemic deterioration, a post hoc subgroup analysis was conducted based on baseline (BL) HbA[sub]1c[/sub] ([lt]8% [n=114], 8% to [lt]9% [n=93], [ge]9% [n=75]). After 18 weeks, adjusted mean reductions from BL HbA[sub]1c[/sub] were greater with SAXA + MET XR vs uptitrated MET XR in the subgroups with BL HbA[sub]1c[/sub] of 8% to [lt]9% (adjusted mean difference [SAXA + MET XR [ndash] uptitrated MET XR] [95% CI]: [minus]0.51 [[minus]0.89, [minus]0.14]) and [ge]9% ([minus]0.86 [[minus]1.28, [minus]0.45]) and numerically greater in the subgroup with BL HbA[sub]1c[/sub] of [lt]8% ([minus]0.30 [[minus]0.63, 0.04]; last observation carried forward [LOCF] analysis). Results (LOCF) were similar for fasting plasma glucose ([lt]8%, [minus]6.07 [[minus]21.86, 9.73]; 8% to [lt]9%, [minus]20.98 [[minus]38.51, [minus]3.45]; [ge]9%, [minus]9.45 [[minus]28.92, 10.02]) and 2-h postprandial glucose ([lt]8%, [minus]10.00 [[minus]33.43, 13.43]; 8% to [lt]9%, [minus]37.10 [[minus]64.63, [minus]9.56]; [ge]9%, [minus]26.54 [[minus]57.15, 4.07]). A greater percentage of patients achieved HbA[sub]1c[/sub] [lt]7% at week 18 [LOCF] with SAXA + MET XR vs uptitrated MET XR in the BL HbA[sub]1c[/sub] [lt]8% subgroup (73.5% vs 46.2%; SAXA + MET XR [minus] uptitrated MET XR [95% CI], 27.3% [8.7%, 43.7%]) and a numerically higher percentage achieved HbA[sub]1c[/sub] [lt]7% in the BL HbA[sub]1c[/sub] 8% to [lt]9% subgroup (25.0% vs 11.6%; 13.4% [[minus]3.4%, 29.6%]). Few patients with BL HbA[sub]1c[/sub] [ge]9% achieved HbA[sub]1c[/sub] [lt]7% (7.5% vs 5.9%; 1.6 [[minus]13.6, 15.6]). [italic]P[/italic] values for a test of consistency of treatment effect by BL HbA[sub]1c[/sub] were [italic]P[/italic]=0.13 for HbA[sub]1c[/sub], [italic]P[/italic]=0.84 for fasting plasma glucose, and [italic]P[/italic]=0.28 for postprandial glucose. With SAXA + MET XR vs uptitrated MET XR, rates of [ge]1 adverse event (AE; lowest to highest BL HbA[sub]1c[/sub] subgroups, respectively) were 51.0% vs 52.3%, 49.0% vs 43.2%, and 57.5% vs 45.7%; rates of [ge]1 serious AE were 0 vs 3.1%, 0 vs 0, and 0 vs 2.9%; and rates of discontinuation owing to AEs were 0% vs 4.6%, 0% vs 0%, and 2.5% vs 8.6%. No treatment-related serious AEs occurred during the study. Adding SAXA 5 mg to MET XR 1500 mg/d was well tolerated and demonstrated numerically greater reduction in HbA[sub]1c[/sub] vs uptitrated MET XR 2000 mg/d independent of BL HbA[sub]1c[/sub].[br][br]Sources of Research Support: This study was supported by Bristol-Myers Squibb and AstraZeneca. Medical writing support for the preparation of this abstract was provided by Complete Healthcare Communications, Inc., with funding from Bristol-Myers Squibb and AstraZeneca.[br][br]Disclosures: BH: Employee, Astra Zeneca. VF: Researcher, GlaxoSmithKline, Novo Nordisk, Novartis Pharmaceuticals, Takeda, Eli Lilly & Company, Astra Zeneca, Sanofi-Aventis, Daiichi-Sankyo Pamlabs; Consultant, GlaxoSmithKline Novartis Pharmaceuticals, Takeda, Novo Nordisk, Sanofi-Aventis, Eli Lilly & Company, Daiichi-Sankyo Pamlabs; Speaker, GlaxoSmithKline, Novartis Pharmaceuticals, Takeda, Novo Nordisk, Sanofi-Aventis, Eli Lilly & Company, Daiichi-Sankyo, Pamlabs. GM: Contractor, Astra Zeneca. MD: Employee, Bristol-Myers Squibb. PO: Employee, Astra Zeneca. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1783 300 2113 MON-192 PO11-01 Monday 1781 2012


1778 ENDO12L_MON-193 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Frequency of Hypoglycemia: Patients Receiving Saxagliptin Plus Metformin vs. Sulfonylurea (SU) Plus Metformin in Real-World Clinical Practice Suellen M Curkendall, Bin Zhang, Gregory Lenhart, Erin E Thomson, Kelly F Bell, John P Graham Thomson Reuters, Washington, DC; Bristol-Myers Squibb, Plainsboro, NJ A randomized clinical trial comparing saxagliptin (a DPP-4 inhibitor) with glipizide (a sulfonylurea (SU)) as add-on therapy to metformin in patients with T2DM found that although both regimens had similar efficacy in improving glycemic control, the rate of hypoglycemia was much lower in the saxagliptin group. The current study used healthcare claims data to compare the rates of hypoglycemia in actual clinical practice in patients with T2DM on metformin who initiated treatment with saxagliptin vs. SU.[br]This was a retrospective analysis of commercially and Medicare-insured adults in the U.S. who were on metformin and added SU or saxagliptin from 08/01/2009-12/31/2010. Patients were excluded if they had any prescriptions for insulin or non-insulin antidiabetic drugs other than metformin and the study drugs (saxagliptin or SU) during the 6 month pre-period. A hypoglycemia event was defined as a diagnosis of hypoglycemia on an outpatient or emergency room insurance claim or principal diagnosis on a hospital claim or a glucagon injection in the outpatient setting. The rates of hypoglycemia (measured as per 100 person-years [PY]) were compared during 4 months following the initiation of saxagliptin or SU. To control for potential confounding, SU patients were matched to saxagliptin patients (5 to 1) using propensity scores and the rate ratio was further adjusted using a multivariate model.[br]A total of 1,567 and 21,025 patients qualified for the saxagliptin and SU cohorts, respectively. The saxagliptin patients were matched to 7,835 SU patients. The rate of hypoglycemia in the saxagliptin cohort was 1.74 per 100 PY vs. 6.68 in the SU cohort (rate ratio 0.31, 95% CI: 0.14 [ndash] 0.60) and 4.45 per 100 PY in the matched SU patients (rate ratio 0.39, 95% CI: 0.17 [ndash] 0.77). After multivariate adjustment, the rate ratio was 0.37, (95% CI: 0.19 [ndash] 0.74).[br]Analysis of this claims data demonstrated that saxagliptin had a lower risk of hypoglycemia than SU in patients with T2DM receiving metformin.[br][br]Disclosures: BZ: Employee, Bristol-Myers Squibb. KFB: Employee, Bristol-Myers Squibb. JPG: Employee, Bristol-Myers Squibb. Nothing to Disclose: SMC, GL, EET 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 737 300 2114 MON-193 PO11-01 Monday 1782 2012


1779 ENDO12L_MON-194 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Measures of [beta]-Cell Function and Insulin Sensitivity over Time in Patients with Type 2 Diabetes Receiving Dapagliflozin Versus Glipizide as Add-On Therapy to Metformin Katja Rohwedder, Anna Maria Langkilde, Nayyar Iqbal, Lisa Ying, Afshin Salsali AstraZeneca, Wedel, Germany; AstraZeneca, M[ouml]lndal, Sweden; Bristol-Myers Squibb, Princeton, NJ Dapagliflozin (DAPA), a selective inhibitor of sodium glucose cotransporter 2 (SGLT2), reduces plasma glucose by increasing renal glucose excretion. In this 52-wk study (N=814 [636 completers]) with a 52-wk extension (N=624 [432 completers]), patients with type 2 diabetes (mean baseline HbA1c=7.7%, diabetes duration=6.3 y) inadequately controlled with metformin (1500[ndash]2500 mg/d) received DAPA ([le]10 mg/d) or glipizide (GLIP, [le]20 mg/d) as add-on therapy (NCT00660907). The primary endpoint, adjusted mean change from baseline in HbA1c at wk 52, was identical with DAPA and GLIP, [ndash]0.52%. In this analysis, [beta]-cell function and insulin sensitivity (IS) were assessed by fasting insulin, proinsulin, and C-peptide levels and by homeostatic model assessment (HOMA-2), which has not been validated in the setting of urinary glucose loss with SGLT2 inhibitors. Adjusted mean change from baseline (95% CI) in fasting insulin, proinsulin, C-peptide, HOMA-2 [beta] (%[beta]) and HOMA-2 IS, was evaluated at wk 26, 52, 78 and 104 using longitudinal repeated measures analysis. At 104 wk, mean HbA1c was reduced more with DAPA, [ndash]0.32% ([ndash]0.42%, [ndash]0.21%) than with GLIP, [ndash]0.14% ([ndash]0.25%, [ndash]0.03%). Mean changes in fasting plasma glucose for DAPA and GLIP were [ndash]23.1 ([ndash]26.6, [ndash]19.9) and [ndash]19.0 ([ndash]22.3, [ndash]15.8) mg/dL at wk 52 and [ndash]20.2 ([ndash]23.8, [ndash]16.5) and [ndash]12.2 ([ndash]15.9, [ndash]8.5) mg/dL at wk 104, respectively. Fasting insulin decreased with DAPA at wk 52, [ndash]1.64 ([ndash]2.47, [ndash]0.82) and 104, [ndash]2.35 ([ndash]3.86, [ndash]0.85) [mu]IU/L, but increased with GLIP, 1.28 (0.44, 2.12) and 1.16 ([ndash]0.42, 2.74) [mu]IU/L. Proinsulin decreased at wk 52, [ndash]5.94 ([ndash]7.89, [ndash]3.99) and 104, [ndash]5.46 ([ndash]7.52, [ndash]3.39) pmol/L with DAPA and increased with GLIP, 5.32 (3.35, 7.29) and 4.74 (2.61, 6.88) pmol/L. C-peptide did not change with DAPA at wk 52, [ndash]0.08 ([ndash]0.20, 0.03) or wk 104, [ndash]0.14 ([ndash]0.28, [ndash]0.01) ng/mL, but increased with GLIP, 0.48 ([ndash]0.36, 0.59) and 0.35 (0.21, 0.49) ng/mL. HOMA-2 %[beta] increased more with GLIP, 28.7% (24.5%, 33.0%) than with DAPA, 15.0% (10.8%, 19.2%) at wk 52 and at wk 104,18.0% (13.3%, 22.8%) for GLIP vs 11.6% (7.1%, 16.2%) for DAPA. GLIP decreased HOMA-2 IS at wk 52, [ndash]4.0% ([ndash]6.0%, [ndash]2.0%) and 104, [ndash]1.4% ([ndash]4.4%, 1.6%), whereas DAPA increased HOMA-2 IS at wk 52, 6.0% (4.0%, 8.0%) and 104, 6.5% (3.6%, 9.4%). In conclusion, DAPA showed a greater durability of glycemic control than GLIP and lowered fasting proinsulin. In addition, DAPA led to improvement of IS and [beta]-cell function, whereas GLIP showed worsening of IS but improvement in [beta]-cell function.[br][br]Sources of Research Support: This research was funded by Bristol-Myers Squibb and AstraZeneca. Editorial support was provided by Richard Edwards, PhD, of Complete Healthcare Communications, Inc.[br][br]Disclosures: KR: Employee, Astra Zeneca. AML: Employee, Astra Zeneca. NI: Employee, Bristol-Myers Squibb. LY: Employee, Bristol-Myers Squibb. AS: Employee, Bristol-Myers Squibb. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1672 300 2115 MON-194 PO11-01 Monday 1783 2012


1780 ENDO12L_MON-195 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Efficacy and Safety of Saxagliptin Added to Metformin Extended-Release (XR) Versus Uptitrating Metformin XR in Patients with Type 2 Diabetes: Outcomes in Hispanic Patients Boaz Hirshberg, Vivian Fonseca, Gianmaria Minervini, Mark Donovan, Peter Ohman AstraZeneca, Wilmington, DE; Tulane University Health Sciences Center, New Orleans, LA; Bristol-Myers Squibb, Princeton, NJ Patients with type 2 diabetes not achieving glycemic control on open-label metformin XR 1500 mg/d were randomized to receive 18 weeks of double-blind saxagliptin 5 mg/d added to existing therapy or uptitration of metformin XR to 2000 mg/d (NCT00960076). Patients receiving saxagliptin (n=138) had significantly better improvements in HbA[sub]1c[/sub] than those uptitrating metformin XR (n=134; [italic]P[/italic][lt]0.0001) and treatment was well tolerated. To investigate outcomes in Hispanic patients, a post hoc analysis assessed efficacy, tolerability, and safety in this subgroup using last observation carried forward methodology. Baseline HbA[sub]1c[/sub] values for the saxagliptin group (n=84) and the uptitrated metformin XR group (n=88) were 8.41% and 8.28%, respectively; adjusted mean changes from baseline were -0.94% and -0.55%, respectively (difference: -0.38%; 95% CI: -0.63, -0.14). Differences in adjusted mean reductions in fasting plasma glucose and 2-h postprandial glucose for saxagliptin vs uptitrated metformin were -6.89 mg/dL (95% CI: -16.62, 2.85) and -18.47 mg/dL (95% CI: -34.21, -2.73), respectively. Numerically more Hispanic patients achieved HbA[sub]1c[/sub] [lt]7% with saxagliptin (35.7%) than uptitrated metformin XR (29.1%; difference: 6.6%; 95% CI: -8.0, 20.6). Incidences of any adverse event (AE) were similar in the saxagliptin (52.4%) and uptitrated metformin XR groups (44.7%); whereas 0 vs 4.5%, respectively, discontinued owing to AEs. Similar to the full study, this analysis suggests that in Hispanic patients with type 2 diabetes, saxagliptin 5 mg/d added to metformin XR is more effective than uptitrating metformin XR and is well tolerated.[br][br]Sources of Research Support: This study was supported by Bristol-Myers Squibb and AstraZeneca. Medical writing support for the preparation of this abstract was provided by Complete Healthcare Communications, Inc., with funding from Bristol-Myers Squibb and AstraZeneca.[br][br]Disclosures: BH: Employee, Astra Zeneca. VF: Researcher, GlaxoSmithKline, Novo Nordisk, Novartis Pharmaceuticals, Takeda, Eli Lilly & Company, Astra Zeneca, Sanofi-Aventis, Daiichi-Sankyo, Pamlabs; Consultant, GlaxoSmithKline, Novartis Pharmaceuticals, Takeda, Novo Nordisk, Sanofi-Aventis, Eli Lilly & Company, Daiichi-Sankyo, Pamlabs; Speaker, GlaxoSmithKline, Novartis Pharmaceuticals, Takeda, Novo Nordisk, Sanofi-Aventis, Eli Lilly & Company, Daiichi-Sankyo, Pamlabs. GM: Contractor, Astra Zeneca. MD: Employee, Bristol-Myers Squibb. PO: Employee, Astra Zeneca. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1784 300 2116 MON-195 PO11-01 Monday 1784 2012


1781 ENDO12L_MON-196 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Saxagliptin in Combination with Insulin for Type 2 Diabetes: Outcomes Stratified by Baseline HbA[sub]1c[/sub] and Body Mass Index Gianmaria Minervini, Anthony H Barnett, Bernard Charbonnel, John Monyak, Nayyar Iqbal AstraZeneca, Wilmington, DE; University of Birmingham and Heart of England National Health Science Foundation Trust, Birmingham, UK; Centre Hospitalier Universitaire de Nantes, Nantes, France; AstraZeneca, Wilmington, DE; Bristol-Myers Squibb, Princeton, NJ In a 24-wk placebo (PBO)-controlled phase 3b trial in patients with type 2 diabetes and inadequate response to insulin[plusmn]metformin, adding saxagliptin (SAXA) 5 mg/d statistically significantly improved glycemic control ([italic]P[/italic][lt]0.01) and was well tolerated (NCT00757588). To determine if treatment response varies with disease severity or weight, groups were stratified by baseline (BL) HbA[sub]1c[/sub] ([lt]8% [n=114], 8% to [lt]9% [n=191], [ge]9% [n=150]) and BMI (obese, [ge]30 kg/m[sup]2[/sup] [n=286]; nonobese, [lt]30 kg/m[sup]2 [/sup][n=169]) and analyzed by last observation carried forward. Changes from BL glycemic measures are presented as adjusted mean difference, SAXA[minus]PBO [95% CI] with [italic]P [/italic]values for consistency of treatment effect across subgroups. Reduction from BL HbA[sub]1c[/sub] at wk 24 was greater with SAXA vs PBO and similar across all BL HbA[sub]1c [/sub]categories ([lt]8%, [minus]0.41 [[minus]0.77, [minus]0.06]; 8% to [lt]9%, [minus]0.40 [[minus]0.66, [minus]0.13]; [ge]9%, [minus]0.42 [[minus]0.73, [minus]0.12];[italic] P[/italic]=0.99). Results generally showed numerically greater reductions with SAXA vs PBO for fasting plasma glucose (FPG [mg/dL]; [lt]8%, 3.04 [[minus]20.13, 26.21]; 8% to [lt]9%, [minus]2.70 [[minus]20.53, 15.12]; [ge]9%, [minus]10.86 [[minus]31.08, 9.37]; [italic]P[/italic]=0.75) and 2-h postprandial glucose (PPG [mg/dL]; [lt]8%, [minus]0.13 [[minus]35.28, 35.03]; 8% to [lt]9%, [minus]43.73 [[minus]69.30, [minus]18.16]; [ge]9%, [minus]0.89 [[minus]31.06, 29.27]; [italic]P[/italic]=0.49). More patients achieved HbA[sub]1c[/sub] [lt]7% with SAXA vs PBO in patients with BL HbA[sub]1c[/sub] [lt]8% (difference SAXA vs PBO, 23.0% [7.3%, 38.6%]) and 8% to [lt]9% (11.9% [2.9%, 20.9%]); few patients with BL HbA[sub]1c[/sub] [ge]9% achieved HbA[sub]1c[/sub][lt]7% (actual, 2.0% vs 2.1% [SAXA vs PBO]; difference [95% CI], [minus]0.1% [[minus]5.0%, 4.7%]). Across HbA[sub]1c[/sub] categories, rates of adverse events (AEs; SAXA vs PBO) were similar overall ([lt]8%: 48.7% vs 63.2%; 8% to [lt]9%: 57.1% vs 55.4%; [ge]9%: 62.7% vs 62.5%). In obese and nonobese patients, reduction in adjusted mean HbA[sub]1c[/sub] from BL at wk 24 was greater with SAXA vs PBO (obese, [minus]0.35 [[minus]0.57, [minus]0.12]; nonobese, [minus]0.50 [[minus]0.78, [minus]0.23];[italic] P[/italic]=0.39). Improvement was numerically greater with SAXA vs PBO for reduction from BL FPG (obese: [minus]0.31 mg/dL [[minus]12.22, 11.60]; nonobese: [minus]11.02 mg/dL [[minus]25.92, 3.88]; [italic]P[/italic]=0.27) and 2-h PPG (obese: [minus]25.80 mg/dL [[minus]44.43, [minus]7.17]; nonobese: [minus]19.15 mg/dL [[minus]41.49, 3.19]; [italic]P[/italic]=0.65) and percentage achieving HbA[sub]1c[/sub][lt]7% (obese: 10.2% [2.7%, 17.7%]; nonobese: 11.4% [1.8%, 21.0%]). Rates of AEs (SAXA vs PBO) were similar in obese (52.6% vs 56.7%) and nonobese (64.8% vs 63.9%) patients. SAXA 5 mg/d added to insulin[plusmn]metformin improves glycemic control across categories of BL HbA[sub]1c [/sub]and BMI and is well tolerated.[br][br]Sources of Research Support: Bristol-Myers Squibb and AstraZeneca. Medical writing support for the preparation of this abstract was provided by Complete Healthcare Communications, Inc., with funding from Bristol-Myers Squibb and AstraZeneca.[br][br]Disclosures: GM: Contractor, Astra Zeneca. AHB: Researcher, Bristol-Myers Squibb, Astra Zeneca, Novartis Pharmaceuticals, Boehringer Ingelheim, Eli Lilly & Company, Novo Nordisk, Sanofi-Aventis, Roche Pharmaceuticals, Takeda, GlaxoSmithKline, MSD; Advisory Group Member, Bristol-Myers Squibb, Astra Zeneca, Novartis Pharmaceuticals, Boehringer Ingelheim, Eli Lilly & Company, Novo Nordisk, Sanofi-Aventis, Roche Pharmaceuticals, Takeda, GlaxoSmithKline, MSD; Speaker, Bristol-Myers Squibb, Astra Zeneca, Novartis Pharmaceuticals, Boehringer Ingelheim, Eli Lilly & Company, Novo Nordisk, Sanofi-Aventis, Roche Pharmaceuticals, Takeda, GlaxoSmithKline, MSD. BC: Researcher, Bristol-Myers Squibb, Astra Zeneca; Advisory Group Member, Bristol-Myers Squibb, Astra Zeneca, Boehringer Ingelheim, GlaxoSmithKline, Merck & Co., Novartis Pharmaceuticals, Novo Nordisk, Roche Pharmaceuticals, Sanofi-Aventis, Takeda; Speaker Bureau Member, Bristol-Myers Squibb, Astra Zeneca, Boehringer Ingelheim , GlaxoSmithKline, Merck & Co., Novartis Pharmaceuticals, Novo Nordisk, Roche Pharmaceuticals, Sanofi-Aventis, Takeda. JM: Employee, Astra Zeneca. NI: Employee, Bristol-Myers Squibb. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1764 300 2117 MON-196 PO11-01 Monday 1785 2012


1782 ENDO12L_MON-197 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) The Gonadal Hormone Regulates Plasma Lactate Levels in Type 2 Diabetes Treated with and without Metformin Fang Liu, Ying Shen, Junling Tang, Taishan Zheng, Weiping Jia Shanghai Jiaotong University Affiliated Sixth People[apos]s Hospital, Shanghai Clinical Medical Center of Diabetes, Shanghai Institute for Diabetes, Shanghai Key Laboratory of Diabetes, Shanghai, China; Soochow Unversity Medical School, Soochow, China [bold]Objective[/bold] Our previous study showed there was gender difference of plasma lactate concentrations in type 2 diabetes. This study is to investigate the effect of sex hormone levels on plasma lactic acid levels in type 2 diabetes with and without metformin therapy. [bold]Methods[/bold] The fasting whole blood of 392 type 2 diabetic patients including metformin group (n=199) and non-metformin treatment group (n=193) were collected. Lactic acid(LA) was measured with enzyme-electrode assay. The sexogen levels including testosterone(T) and estradiol(E2) were measured with chemiluminescence microparticle immunoassay, Spearman or Pearson correlation and logistic regression analysis were performed for the associated factors of LA. [bold]Results[/bold] The LA level in metformin group was significantly higher than non-metformin group [(1.26[plusmn]0.43 vs. 1.14[plusmn]0.49) mmol/L,[italic] P[/italic][lt]0.01], and LA levels of females were significantly higher than those of males ([italic]P[/italic][lt]0.01). LA concentrations were positively correlated with E2, but negatively correlated with metformin and T levels ([italic]P[/italic][lt]0.01). Multivariate logistic regression analysis showed that gender, Creatinine(Cr), E2, metformin and T were independent influence factors to lactate levels. The analysis of subgroups demonstrated that the LA concentrations increased with the elevating of E2 ([italic]P[/italic][lt]0.05), but decreased with the rising of T ([italic]P[/italic][lt]0.05). [bold]Conclusions[/bold] Sex hormones play an important role on regulating plasma lactate levels in diabetic patients treated with metformin. The estradiol up-regulate, but testosterone tend to down-regulate lactate levels.[br][br]Sources of Research Support: NSFC grant 81070650.[br][br]Nothing to Disclose: FL, YS, JT, TZ, WJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1076 300 2118 MON-197 PO11-01 Monday 1786 2012


1783 ENDO12L_MON-198 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Cancer Risk in Diabetes Type 2 Patients Receiving Metformin. Systematic Review and Meta-Analysis Diego Espinoza-Peralta, Linda Liliana Munoz-Hernandez, Marcela Rodriguez-Flores, Liliana Velasco-Hidalgo, Francisco Javier Gomez-Perez Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Mexico City, Mexico; Instituto Nacional de Pediatria, Mexico City, Mexico Background: In the last years, is has been suggested that the regular use of metformin in diabetic patients is associated with lower cancer rates as compared with patients who do not receive this drug.[br]Objective: To evaluate the effect of the administration of metformin on the risk of any type of cancer, in patients with type 2 diabetes.[br]Search strategy: We performed a thorough search and meta-analysis of the following databases without time or date restrictions until July 2011 with MeSH terms [ldquo]Diabetes Mellitus, Type 2[rdquo], [ldquo]Therapy[rdquo], [ldquo]Metformin[rdquo] and [ldquo]Neoplasms[rdquo] and DeCS terms [ldquo]Diabetes mellitus tipo 2[rdquo], [ldquo]Metformina[rdquo], [ldquo]Neoplasias[rdquo] and [ldquo]Terapeutica[rdquo] for Latin American databases: Cochrane library, MEDLINE, Highwire Press, Scielo, Lilacs, Artemisa and Pubgle.[br]Selection criteria: We selected cohorts and nested case-control studies which evaluated the risk of cancer in type 2 diabetic patients taking metformin without any other condition that increased the risk of cancer. We did not find any clinical trial that evaluated this specific investigation question.[br]Data collection and information analysis: Two investigators independently selected the studies, evaluated quality using STROBE Statement, extracted data and obtained information for risk estimation for analysis with GRADE process for evidence synthesis.[br]Results: Seven studies were selected for relevance that reported risk in cancer incidence because information quality, reporting 32,405 patients. The use of metformin is associated with a risk reduction of all cancer (Random effect model: OR 0.62, CI 95% 0.54-0.70), and in a secondary analysis less risk in colon (Fixed effects model OR=0.46, CI 95% 0.29-0.63) and breast cancer (OR 0.58, CI 95% 0.29-0.88) and without effect in pancreatic cancer (Random effect model OR1.04, CI 95% 0.071-2.11), as compared with patients not receiving metformin or those who were taking other drugs for glucose control.[br]Conclusions: The regular use of metformin significantly reduces the risk of cancer, despite the heterogeneity of glucose-lowering drugs with which it was compared in all the different studies that were analyzed.[br][br]Nothing to Disclose: DE-P, LLM-H, MR-F, LV-H, FJG-P 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1129 300 2119 MON-198 PO11-01 Monday 1787 2012


1784 ENDO12L_MON-199 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Insulin Degludec Provides Similar Pharmacokinetic and Pharmacodynamic Responses in Black, White, and Hispanic/Latino Subjects with Type 2 Diabetes Marcus Hompesch, Linda Morrow, Elaine Watkins, Carsten Roepstorff, Henrik F Thomsen, Hanne Haahr Profil[reg] Institute for Clinical Research, Inc, Chula Vista, CA; Novo Nordisk A/S, Soeborg, Denmark; Novo Nordisk A/S, Aalborg, Denmark [italic]Objective[/italic]: Insulin degludec (IDeg) is a new-generation basal insulin that forms soluble multi-hexamers upon subcutaneous injection, resulting in an ultra-long action profile. This randomized, double-blind, two-period, cross-over trial investigated the pharmacokinetic and pharmacodynamic properties of IDeg at steady-state (SS).[br][italic]Methods:[/italic] The trial included 63 insulin-treated subjects (26 females and 37 males) with type 2 diabetes of different race and/or ethnicity (18 Black, 23 White, 22 Hispanic/Latino; mean age: 52 yrs, BMI: 32.4 kg/m[sup]2[/sup], A[sub]1c[/sub]: 8.2%). Subjects completed two 6-day treatment periods with 0.6 U/kg/day of IDeg or insulin detemir (IDet). At the end of each treatment period a 24-h euglycemic clamp was performed (Biostator; clamp blood glucose level 90 mg/dL).[br][italic]Results:[/italic] IDeg resulted in a flat and stable glucose-lowering effect extending beyond 24 h in all subjects across race/ethnic groups. The effect was evenly distributed between the first and second 12 h in all groups (AUC[sub]GIR,0-12h,SS[/sub]/AUC[sub]GIR,[tau],SS[/sub] ranging between 0.48 and 0.54). The total glucose-lowering effect of Ideg at SS (AUC[sub]GIR,[tau],SS[/sub]) was 1940, 1722, and 2286 mg/kg in Black, White, and Hispanic/Latino subjects, respectively. Pair-wise comparisons were not statistically significantly different (mean difference [95% CI]: Black vs. Hispanic -346 [-1088; 396]; Black vs. White 218 [-551; 986]; Hispanic vs. White 564 [-168; 1295] mg/kg). Pair-wise comparisons of total Ideg exposure (AUC[sub]Ideg,[tau],SS[/sub]) were also similar between the three groups with mean ratios [95% CI] of: Black vs. Hispanic 1.13 [0.95; 1.34], Black vs. White 1.10 [0.91; 1.31], and Hispanic vs. White 0.97 [0.82; 1.16] pmol[bull]h/L). There was no statistically significant difference in the race/ethnicity pattern between Ideg and Idet for AUC[sub]GIR,[tau],SS[/sub], p=0.85 or AUC[sub]ins,[tau],SS[/sub], p=0.58. Ideg was well tolerated and no safety issues were identified.[br][italic]Conclusions:[/italic] Ideg resulted in similar pharmacokinetic and pharmacodynamic responses across race and ethnicity in subjects with type 2 diabetes. Thus, the ultra-long pharmacological properties of Ideg are preserved across race and ethnic groups. While insulin doses must be adjusted on an individual basis, this trial supports the notion that differences in the response to Ideg should not be anticipated based on differences in race or ethnicity between patients.[br][br]Disclosures: MH: Employee, Profil® Institute for Clinical Research, Inc. LM: Employee, Profil® Instiute for Clinical Research Inc. EW: Employee, Profil® Institute for Clinical Research Inc. CR: Employee, Novo Nordisk. HFT: Employee, Novo Nordisk. HH: Employee, Novo Nordisk. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 572 300 2120 MON-199 PO11-01 Monday 1788 2012


1785 ENDO12L_MON-200 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Effects of Pioglitazone on Bone in Postmenopausal Women with Impaired Fasting Glucose or Impaired Glucose Tolerance Henry Bone, Robert Lindsay, Michael McClung, Alfonso Perez, Marsha Raanan, Robert Spanheimer Michigan Bone and Mineral Clinic, Detroit, MI; Helen Hayes Hospital, West Haverstraw, NY; Oregon Osteoporosis Center, Portland, OR; Takeda Global Research [amp] Development Center, Inc, Deerfield, IL; Takeda Pharmaceuticals North America, Inc, Deerfield, IL [bold]Background:[/bold] Meta-analyses of clinical studies have shown an increased incidence of peripheral fractures in women with type 2 diabetes mellitus (T2DM) taking pioglitazone (Pio), although the mechanism of this increase is not known.[br][bold]Purpose:[/bold] We assessed the effect of Pio on bone mineral density (BMD) and turnover in postmenopausal women with impaired fasting glucose (IFG) or impaired glucose tolerance (IGT).[br][bold]Methods:[/bold] A total of 156 postmenopausal women [le]70 years old (mean, 60 years) with documented IFG or IGT were randomized to receive Pio 30 mg QD (n=78), titrated to 45 mg QD after 1 month, or placebo (n=78) for 12 months, followed by a 6-month washout/follow-up. Participants took vitamin D and calcium supplements throughout the 18-month period. BMD measurements at baseline, 12 months, and 18 months included total proximal femur (primary endpoint), total body, femoral neck, lumbar spine, and radius by dual-energy x-ray absorptiometry. Percentage changes from baseline were compared between treatment groups using one-way analysis of covariance with baseline value as a covariate.[br][bold]Results:[/bold] Least square mean changes to month 12 in total proximal femur BMD were -0.69% for Pio and -0.14% for placebo (p=0.170); no statistically significant differences between Pio and placebo were seen in change from baseline to month 12 or from month 12 to 18 in any BMD measures. Bone remodeling markers (bone-specific alkaline phosphatase, osteocalcin, procollagen type I N-terminal propeptide, serum [beta]-C-terminal telopeptide of type I collagen, and urine N-telopeptide of type I collagen) demonstrated no significant effects of Pio compared with placebo. An increase in body weight was seen with Pio at month 12 (1.4 kg vs a decrease of 0.74 kg with placebo; p=0.003), largely accounted for by an increase in total body fat. Fractures confirmed through central adjudication occurred in one Pio and three placebo participants. Over the 18-month period, 1 Pio participant (1.3%) and 8 placebo participants (10.3%) progressed to overt T2DM based on American Diabetes Association criteria.[br][bold]Conclusions:[/bold] This study showed no evidence of BMD loss or altered bone metabolism in postmenopausal women with IFG or IGT receiving high-dose Pio, while demonstrating expected effects of Pio on weight and measures of glycemic control.[br][br]Sources of Research Support: Takeda Global Research and Development Center, Inc.[br][br]HB: Consultant, Amgen, Merck & Co, Takeda, Tarsa; Investigator, Amgen, Merck & Co., Nordic Bioscience A/S, Novartis Pharmaceuticals, Takeda, Tarsa; Speaker Bureau Member, Amgen. RL: Speaker, Lilly USA, LLC, Amgen, Novartis Pharmaceuticals; Investigator, Takeda; Consultant, Takeda, Lilly USA, LLC, Amgen. MM: Investigator, Amgen, Merck & Co., Takeda; Consultant, Amgen, USA, LLC, Merck & Co., Novartis Pharmaceuticals, Warner Chilcott Pharmaceuticals, Takeda; Speaker, Amgen, Lilly USA, LLC, Merck & Co., Novartis Pharmaceuticals, Warner Chilcott Pharmaceuticals. AP: Employee, Takeda. MR: Employee, Takeda. RS: Employee, Takeda. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1727 300 2121 MON-200 PO11-01 Monday 1789 2012


1786 ENDO12L_MON-201 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Insulin Degludec Does Not Increase Antibody Formation Compared to Insulin Glargine: An Evaluation of Phase 3a Clinical Trials Priscilla Hollander, Jiten Vora, Soren Can Tamer, Thue Johansen, Richard M Bergenstal Baylor Endocrine Center, Dallas, TX; International Diabetes Center, Minneapolis, MN; Novo Nordisk A/S, Soeborg, Denmark; Novo Nordisk A/S, Soeborg, Denmark; Royal Liverpool University Hospitals, Liverpool, UK [bold]Background[/bold][br]Insulin degludec (IDeg) is an ultra-long-acting basal insulin analog where the threonine in position B30 of human insulin has been omitted and the lysine in position B29 has been coupled to hexadecanedioic acid via a glutamic acid spacer. IDeg forms soluble multi-hexamers upon subcutaneous injection, resulting in a stable and consistent glucose-lowering effect. All insulin preparations cause some degree of antibody formation in humans, but this has little or no effect on the efficacy or safety of currently marketed products. Here we present antibody data from all phase 3a clinical trials in patients with type 1 (T1DM) or type 2 (T2DM) diabetes where insulin antibody development was analyzed for IDeg and compared to insulin glargine (IGlar).[br][bold]Methods[/bold][br]Insulin antibodies were measured in 6 randomized, controlled, open-label, treat-to-target trials (26[ndash]52 weeks duration): 2 trials in T1DM (IDeg: n=801; IGlar: n=315) and 4 trials in T2DM (IDeg: n=1734; IGlar: n=860). In all trials, IDeg and IGlar were dosed once daily. Insulin aspart was used as bolus insulin in the T1DM trials. Antibody levels (specific and cross-reacting to human insulin) were measured using specific radioimmunoassays and expressed as percent bound/total radioactivity (%B/T).[br][bold]Results[/bold][br]Across all trials, very few subjects developed IDeg- and IGlar-specific antibodies and only at very low levels. Furthermore, mean levels of antibodies cross-reacting to human insulin remained low for both groups throughout the duration of the trials for subjects with T1DM (10[ndash]15% B/T) and T2DM ([lt]6% B/T). For both IDeg and IGlar, Spearman correlation coefficients were calculated on an individual trial basis to investigate the influence of cross-reacting antibodies on change from baseline in HbA[sub]1c[/sub], and HbA[sub]1c[/sub] and total daily insulin dose at end-of-trial. All correlation coefficients were low indicating that there was no clinically relevant influence of insulin antibodies on HbA[sub]1c[/sub] or dose. Based on scatter plots, there was no difference in HbA[sub]1c[/sub] or total insulin dose at end-of-trial between subjects having cross-reacting antibody levels of [gt]10% B/T and those with levels of [lt]10% B/T. The adverse event profile in subjects with an antibody titer of [gt]10% B/T was generally similar to that of the overall trial population.[br][bold]Conclusion[/bold][br]The immunogenic responses to long-term treatment with insulin degludec and insulin glargine are similar and low, with no clinically relevant impact on HbA[sub]1c[/sub] or total daily insulin dose.[br][br]Disclosures: PH: Advisory Group Member, Novo Nordisk; Medical Advisory Board Member, Novo Nordisk; Researcher, Novo Nordisk. JV: Advisory Group Member, Novo Nordisk; Speaker Bureau Member, Novo Nordisk; Researcher, Novo Nordisk. SCT: Employee, Novo Nordisk. TJ: Employee, Novo Nordisk. Nothing to Disclose: RMB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 578 300 2122 MON-201 PO11-01 Monday 1790 2012


1787 ENDO12L_MON-202 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Effect of Pioglitazone Monotherapy on Body Composition among Asian Indian T2DM Patients Prashant Mathur, Poonam Punjabi, Mukul Mathur, Deepak Kumar Gupta, Jyoti Thanvi, Sandeep Kumar Mathur SMS Medical College, Jaipur, India; SMS Medical College, Jaipur, India; SMS Medical College, Jaipur, India; Jaipur College of Pharmacy, Jaipur, India; Central University of Rajasthan, Kishangarh, India; SMS Medical College, Jaipur, India [bold]AIM:[/bold][br]To study effect of pioglitazone mono-therapy on body composition in treatment naive Asian Indian T2DM patients.[br][bold]METHODS:[/bold][br]Thirty seven newly diagnosed T2DM patients (age 49.70 [plusmn] 9.62 yrs, M: F ratio 27:10) were treated with Pioglitazone 30 mg PO once daily for at least 6 months. Inclusion criteria were BMI[lt]30 and no contraindication to the drug. Changes in following parameters were compared at start and after 6 months of therapy: Plasma glucose, Lipid profile, BMI, HOMA-R, HOMA-B and body composition measured by dual-energy X-ray absorptiometry. Relationship between changes in parameters of body composition and changes in plasma glucose, insulin resistance, lipid profile and BMI after the therapy was studied.[br][bold]RESULTS:[/bold][br]Pioglitazone significantly decreased HbA1c (10.40 [plusmn] 2.02 to 7.41 [plusmn] 1.29), FPG (214.67 [plusmn] 56.61 to 122.67 [plusmn] 31.16), PPG (310.40 [plusmn] 79.44 to 180.13 [plusmn] 58.84), insulin resistance (2.60 [plusmn] 1.96 to 1.27 [plusmn] 1.01) (p[lt]0.001) and triglycerides HDL ratio (4.14 [plusmn] 2.29 to 3.30 [plusmn] 1.63). A significant increase in BMI (23.82 [plusmn] 2.81 to 25.01 [plusmn] 3.13), and HOMA-B (14.07 [plusmn] 11.66 to 29.81 [plusmn] 22.93) was observed. A significant increase (p[lt]0.001) in the fat content of all limbs and total body fat was observed. But there was no statistically significant change in the trunk composition. However head fat and lean mass increased significantly.[br]Increase in limb fat was found to be associated with decrease in FPG (p[lt]0.01), PPG (p[lt]0.01), insulin resistance (p[lt]0.01) and increase in BMI (p[lt]0.05). A significant association (p[lt]0.05) was found between decrease in triglyceride HDL ratio and increase in fat, lean [amp] total mass of head. No correlation was found between increase in beta cell function and increase in fat composition of limbs.[br][bold]CONCLUSIONS:[/bold][br]Pioglitazone alters body composition by increasing limb fat content, without altering truncal fat. Its glycemic response is associated with increase in limb fat. Its lipid response was found to be associated with increase in head fat content.[br][br]Nothing to Disclose: PM, PP, MM, DKG, JT, SKM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1901 300 2123 MON-202 PO11-01 Monday 1791 2012


1788 ENDO12L_MON-203 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Low Cardiovascular (CV) Risk Hazard Ratio Observed with Linagliptin in Type 2 Diabetes: Further Insights from a Predefined CV Meta-Analysis Odd-Erik Johansen, Dietmar Neubacher, Maximilian von Eynatten, Sanjay Patel, Hans-Juergen Woerle Boehringer Ingelheim, Asker, Norway; Boehringer Ingelheim, Biberach, Germany; Boehringer Ingelheim, Ingelheim, Germany; Boehringer Ingelheim, Bracknell, UK [bold]Background[/bold]: Incidence of cardiovascular (CV) events is increased in type 2 diabetes (T2D) but the potential of CV risk modulation with glucose lowering is debated. Linagliptin, a DPP-4 inhibitor, provides a meaningful reduction in hyperglycemia and also may have potential CV benefits[sup]1[/sup]. To further explore potential mechanisms behind this, we report additional relevant data from a large CV meta-analysis of linagliptin trial data.[br][bold]Methods[/bold]: In this prespecified meta-analysis from 8 Phase III randomized, controlled trials, CV events were prospectively adjudicated by a blinded independent expert committee. A composite primary endpoint of CV death, MI, stroke, and hospitalization for unstable angina pectoris (UAP) was used. Results are given as hazard ratio (HR) for time to first occurrence of any components of the primary endpoint using a Cox regression model for linagliptin vs total comparators with factors of study (grouped by design), treatment (linagliptin vs combined comparators), gender (female vs male), race (Asian vs White), and time since T2D diagnosis ([gt]5 vs [le]5 yr). Comparative data for conventional CV risk factors across treatment groups are reported descriptively.[br][bold]Results[/bold]: 3319 patients received linagliptin once daily (5 mg: 3159, 10 mg: 160) and 1920 comparator (placebo: 977, glimepiride: 781, voglibose: 162). The overall study cohort (N=5239) seemed representative of a general T2D population among which overall HR was higher in patients with longer duration of T2D (1.51 [95% CI: 0.75-3.04]) and reduced in females (0.36 [95% CI: 0.16-0.84]) and Asians [0.28 [95% CI: 0.10-0.82]). A significantly reduced risk for the primary endpoint after treatment with linagliptin was indicated by the HR (0.36 [95% CI: 0.17-0.74]). Between-group changes in cholesterol and systolic and diastolic blood pressure were similar for linagliptin- and comparator-treated patients. Linagliptin treatment was associated with a greater reduction in HbA1c (-0.6[plusmn]0.9%), fasting triglycerides (-11[plusmn]136 mg/dL), and no weight gain (0.01[plusmn]3.09 kg) in contrast to the total comparators (respectively -0.3[plusmn]1.0%, -6[plusmn]156 mg/dL, and 0.6[plusmn]3.42 kg).[br][bold]Conclusion[/bold]: Additional Cox regression analyses support the CV safety profile of linagliptin in a large phase III CV meta-analysis. The low CV HR observed with linagliptin cannot be fully explained by classical CV risk factors such as blood pressure and lipids. Further clinical outcome and mechanistic studies are underway to provide additional insights.[br][br](1) Johansen OE, Neubacher D, von Eynatten M, Patel S, Woerle HJ. Cardiovascular safety with linagliptin in patients with type 2 diabetes mellitus: a pre-specified, prospective, and adjudicated meta-analysis of a phase 3 program. Cardiovasc Diabetol. 2012, 11:3 doi:10.1186/1475-2840-11-3.[br][br]Sources of Research Support: Boehringer Ingelheim.[br][br]Disclosures: O-EJ: Employee, Boehringer Ingelheim. DN: Employee, Boehringer Ingelheim. MvE: Employee, Boehringer Ingelheim. SP: Employee, Boehringer Ingelheim. H-JW: Employee, Boehringer Ingelheim. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1973 300 2124 MON-203 PO11-01 Monday 1792 2012


1789 ENDO12L_MON-204 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Elderly Patients Experience a Lower Rate of Nocturnal Hypoglycemia with Insulin Degludec Than with Insulin Glargine: A Meta-Analysis of Phase 3a Trials Christopher H Sorli, Mark L Warren, Henriette Mersebach, Thue Johansen, David S Oyer Billings Clinic, Billings, MT; Physicians East, PA, Greenville, NC; Novo Nordisk A/S, Soeborg, Denmark; Northwestern University Feinberg School of Medicine, Chicago, IL [bold]Background[/bold][br]Choosing diabetes therapies for the elderly can be challenging as they may be particularly vulnerable to hypoglycemia, especially at night. Insulin degludec (IDeg), a new basal insulin that forms soluble multi-hexamers after sc injection, has an ultra-long, stable action profile with low hour-to-hour and day-to-day variability in glucose-lowering effect. IDeg has demonstrated less hypoglycemia than other current basal insulins. The objective of this analysis was to compare hypoglycemia rates with IDeg vs. insulin glargine (IGlar) in subjects aged [ge]65 years.[br][bold]Methods[/bold][br]A pre-planned meta-analysis evaluating patient level data from the IDeg 3a program using a negative binomial regression model was performed in a pooled population with type 1 (T1DM) or type 2 (T2DM) diabetes to compare hypoglycemia rates with IDeg vs. IGlar. Confirmed hypoglycemia was defined as self-reported PG [lt]56 mg/dL or a severe episode requiring assistance; nocturnal confirmed hypoglycemia had onset 00:01-05:59, inclusive. The analysis included patient level data from all open-label, randomized, treat-to-target, confirmatory 26- or 52-week trials with IDeg (n=2899) vs. IGlar (n=1431) dosed once daily in T1DM (2 trials) and T2DM (5 trials). Here we present the pre-planned sub-analysis in subjects aged [ge]65 yrs (IDeg, n=632; IGlar, n=283) using the same model.[br][bold]Results[/bold][br]Across trials, 21.1% of all subjects (6.4% of T1DM and 25.3% of T2DM) were aged [ge]65 years. Consistent with confirmatory analyses in the overall population, elderly subjects had fewer confirmed hypoglycemic episodes with IDeg than IGlar. Confirmed hypoglycemia rates were numerically lower in IDeg-treated elderly subjects (rate ratio (RR) IDeg/IGlar: 0.82 [95% CI: 0.66; 1.00]), approaching statistical significance. Nocturnal hypoglycemia rates were significantly 35% lower with IDeg (RR: 0.65 [95% CI: 0.46; 0.93]).[br][bold]Conclusion[/bold][br]Elderly subjects treated with IDeg experience a numerically lower rate of confirmed hypoglycemia vs. IGlar, consistent with overall population results. Importantly, elderly subjects experienced a significantly 35% lower rate of nocturnal hypoglycemia with IDeg vs. IGlar, confirming what has been seen throughout the IDeg clinical program. The lower rate of hypoglycemia associated with IDeg may be of particular benefit in the elderly population on insulin.[br][br]Disclosures: CHS: Speaker Bureau Member, Novo Nordisk. MLW: Advisory Group Member, Novo Nordisk; Speaker Bureau Member, Novo Nordisk; Research Funding, Novo Nordisk. HM: Employee, Novo Nordisk. TJ: Employee, Novo Nordisk. DSO: Advisory Group Member, Novo Nordisk; Speaker Bureau Member, Novo Nordisk. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 554 300 2125 MON-204 PO11-01 Monday 1793 2012


1790 ENDO12L_MON-205 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Positive Effects of Once Daily Incretin Use in the Very Insulin Resistant JaneFrances Chukwu, Irl Hirsch, Dace Trence University of Washington, Seattle, WA Insulin resistance is defined as the use of over 200 units of insulin daily to control blood glucose. This population can be challenging to manage, due to the high volumes of insulin required. In turn, this large amount of insulin can be associated with patient economic burden as well as difficulty in attaining and maintaining glycemic goals. We present a review of 13 insulin resistant individuals with liraglutide added to their insulin regimen, seen in an academic diabetes care clinic, followed for at least 6 months from liraglutide initiation.[br]As would be expected, the majority of patients (77%) had type 2 diabetes. while the rest type 1 diabetes. Mean weight of the subjects prior to initiation of liraglutide was 128.6kg, range 66.8-181.3kg. Ten individuals experienced weight loss, ranging from 0.45-14kg; mean loss for these patients was 4.5kg. Using all patients in an intention-to-treat analysis, the overall weight loss was significant with P-value of 0.0481.Patients were on an average of 2.3 units/kg of insulin (range 1.2-6.6units/kg) prior to initiation of liraglutide. On follow-up, this decreased to an average of 1.6 units/kg (range 0.8-2.6units/kg). Daily insulin dose specifically decreased in 7 patients, the decreases were significant, ranging from 11 to 60units (P-value of 0.0092). Only one person had a noted impact on blood pressure measurement, none had a noted change in lipid measurements. From a mean starting A1C of 8.9% (range 5.7 to11.1%), the A1C dropped to an average of 7.6% (range 5.6 to 9.0%), (p=0.0159). Using Pearson correlation coefficient, there was no correlation between weight loss and change in A1C (value-0.11378). Daily dose of liraglutide ranged from 1.2 to 1.8mg[br]In conclusion, in those with diabetes with severe insulin resistance, defined as the daily use of over 200 units of insulin, once daily incretin addition was effective in improving glucose control as defined by A1C. This suggests that incretin addition can be effective even in the severely insulin resistant, with decreased daily insulin requirement. Weight loss was noted, which could improve insulin sensitivity, but was limited to an average loss of 4.5 pounds. There was no correlation between weight loss and A1C. Specific mechanisms regarding liraglutide[apos]s beneficial effects in this patient population warrants further study.[br][br]Disclosures: IH: Investigator, Novo Nordisk. DT: Investigator, Novo Nordisk. Nothing to Disclose: JC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1366 300 2126 MON-205 PO11-01 Monday 1794 2012


1791 ENDO12L_MON-206 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Efficacy and Tolerability of Metformin Treatment Are Associated with Previously Unknown Genetic Polymorphism Lauma Zarina, Linda Tarasova, Inga Kempa, Marite Cirse, Liene Nikitina-Zake, Kristine Geldnere, Ilze Konrade, Aivars Lejnieks, Janis Klovins, Valdis Pirags Pauls Stradin[scaron] Clinical University Hospital, Riga, Latvia; Latvian Biomedical Research and Study Center, Riga, Latvia; University of Latvia, Riga, Latvia; Riga Stradin[scaron] University, Riga, Latvia [bold]Background. [/bold]Many patients with type 2 diabetes (T2D) are not reaching glycemic targets even by using different combinations of antidiabetic drugs, including insulin. On the other hand, intensified multidrug therapy is associated with increased risk of severe side effects and individualized selection of medication is essential. The aim of our study is to evaluate efficiency of personalized pharmacotherapy according to the genotype and phenotype of T2D patients.[br][bold]Methods/design. [/bold]95 treatment naive T2D patients were involved in prospective clinical trial. Genotyping by GoldenGate Genotyping Assay with VeraCode technology ([italic]Illumina, Inc[/italic].) of 192 SNP in previously reported genes influencing T2D treatment efficiency and tolerability (OCT1,2,3, MATE1,2, PMAT and others) was performed in all patients.[br][bold]Results[/bold]. First results of treatment efficiency and tolerability are currently available in 61 patients. Decrease of the mean HbA[sub]1c[/sub] from 7.7 [plusmn] 2.0% to 6.7 [plusmn] 0.8% and the mean BMI from 34.6 [plusmn] 5.9 kg/m[sup]2[/sup] to 34.1 [plusmn] 5.6 kg/m[sup]2[/sup] was measured. SNP located in region between genes SLC22A2 - SLC22A3 was associated with HbA[sub]1c[/sub] decrease after 3 month metformin therapy and the commonly carried A allele was more often detected in patients who responded to metformin therapy. Unadjusted association test (P = 9.29e-005, OR (CI 0.95) = 4.971[2.163-11.43]) identified association between genetic polymorphisms and response to metformin therapy. Association maintained statistical significance after Bonferroni correction (P=0.01793).[br][bold]Conclusions[/bold]. Genotyping of large number of known polymorphisms in genes responsible for efficiency of T2D therapy may help to individualize treatment schemes. Genetic polymorphisms in SLC22A2 - SLC22A3 gene region are associated with efficiency of short-term metformin treatment.[br][br]Sources of Research Support: Latvian National Reasearch Program in Biomedicine and Public Health 2009-2013.[br][br]Nothing to Disclose: LZ, LT, IK, MC, LN-Z, KG, IK, AL, JK, VP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1636 300 2127 MON-206 PO11-01 Monday 1795 2012


1792 ENDO12L_MON-207 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) A Retrospective Study Comparing Neutral Protamine Hagedorn Insulin with Glargine as Basal Therapy in Glucocorticoid-Associated Diabetes Mellitus in Hospitalized Patients Subarna M Dhital, Dawn B Davis, Yoram Shenker, Melissa Meredith University of Wisconsin School of Medicine and Public Health, Madison, WI [bold][underline]Objective:[/underline][/bold] To compare glycemic outcomes in subjects receiving Neutral Protamine Hagedorn insulin (NPH) versus glargine as basal insulin for management of hyperglycemia associated with glucocorticoid use in hospitalized patients.[br][bold][underline]Methods:[/underline][/bold] We conducted a retrospective review of electronic medical records in 120 adult subjects with hyperglycemia receiving high dose glucocorticoids in a university hospital. Powered at 90% to detect a difference of 27 mg/dL between the two treatment regimens, sixty consecutive subjects were selected in both the NPH and the glargine cohorts using appropriate inclusion and exclusion criteria. Baseline characteristics were assessed in each cohort. Glycemic outcomes were analyzed by comparing fasting blood glucose (BG), mean daily BG, median daily BG and the number of episodes of hypoglycemia per day. The cohorts were compared using unpaired t-test (with Welch[apos]s correction when variances were unequal).[br][bold][underline]Results:[/underline][/bold] NPH and glargine cohorts were similar with regards to age, sex, race, body mass index, hemoglobin A1c, serum creatinine and dose of glucocorticoids. Both cohorts were on high dose glucocorticoids with a majority on prednisone (62% vs. 73%; p=0.17). The majority of the subjects in both groups were managed by an inpatient diabetes management service ensuring a homogeneous management style. One notable difference in baseline characteristics was that the glargine cohort had significantly more subjects with type 1 diabetes (23% vs. 7%, p= 0.011). Glycemic outcomes were similar with regards to mean fasting BG (139[plusmn]55 mg/dL vs. 147[plusmn]57 mg/dL, p= 0.46), mean daily BG (170[plusmn]47 mg/dL vs. 171[plusmn]52 mg/dL, p= 0.89), median BG (166[plusmn]52 mg/dL vs. 166[plusmn]57 mg/dL, p= 0.99), and number of hypoglycemic episodes per day (0.2[plusmn]0.5 vs. 0.1[plusmn]0.3, p= 0.20). Subgroup analyses also did not reveal any significant difference in glycemic outcomes.[br][bold][underline]Conclusion:[/underline][/bold] NPH is commonly considered for hyperglycemia in patients using prednisone because the pharmacokinetics parallel one another (1). However, we have found that NPH and glargine appear to be equally efficacious as basal insulins in the management of hyperglycemia in hospitalized patients receiving high dose glucocorticoids.[br][br](1) Clore JN et al., Endocr Pract 2009; 15:469.[br][br]Nothing to Disclose: SMD, DBD, YS, MM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 406 300 2128 MON-207 PO11-01 Monday 1796 2012


1793 ENDO12L_MON-208 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Hypoglycemic Events in Hospitalized Patients with Diabetes Iraklii Buziashvili, Stanislaw Piotr Klek, Sukhminder Sahansra, Lawrence Ezra Shapiro Winthrop-University Hospital, Mineola, NY OBJECTIVE: Hypoglycemia is a common complication of insulin treatment in hospitalized patients. Several studies show that number and severity of inpatient hypoglycemic events correlate with hospitalization outcomes. Prediction and prevention of hypoglycemic events may be beneficial in decreasing morbidity, mortality and length of hospital stay. We characterized hypoglycemic events in patients with diabetes and examined whether age, renal or hepatic function, and white blood cell (WBC) count correlate with number and/or severity of hypoglycemic events. DESIGN AND PATIENTS: Retrospective study of 294 hypoglycemic events that occurred in 181 hospitalized patients with type 1 or 2 diabetes (1.62 events per patient) within a 3-month period between 01/01/2011 and 03/31/2011. Hypoglycemic event was defined as a point-of-care blood glucose (POC BG) less than 70 mg/dl. Data regarding age and gender of patients, time of hypoglycemic events, creatinine and albumin levels as well as WBC count were retrieved from hospital electronic medical records. Data are presented as means [plusmn] SD. RESULTS: Average age of patients was 72 [plusmn] 16.7 years, with 62% (183/294) of all hypoglycemic events occurring in patients older than 70 years. Average POC BG was equal to 54 [plusmn] 10.5 mg/dl. 73% (213/294) of hypoglycemic events were classified as mild (POC BG 50-69 mg/dl), 24% (71/294) as moderate (POC BG 30-49 mg/dl), and 3% (10/294) as severe (POC BG less than 30 mg/dl). 113 females had 66% (194/294) of all hypoglycemic events, while 68 males had the remaining 34% (100/294) of events. 71% (209/294) of hypoglycemic events were registered during scheduled times of POC BG measurements, with 29% (85/294) of events occuring before breakfast (7:01-9:00), 16% (47/294) before lunch (11:01-13:00), 14% (41/294) before dinner (16:01-18:00), and 12% (36/294) at bedtime (21:01-23:00). There was no correlation of the severity of hypoglycemia (POC BG) with age, creatinine level or WBC count, while there was a weak, but significant correlation with albumin level (r = 0.125, p [lt] 0.01). Most of the hypoglycemic events occurred in patients with normal creatinine (0.6-1.5 mg/dl, 52%) and albumin (3.1-5.0 mg/dl, 58%) levels, as well as normal WBC count (4.0-10.0 [times] 106 cells/ml, 63%). CONCLUSIONS: Elderly patients with diabetes have the highest risk of hypoglycemic events in the hospital. Most of the events are mild and occur in the morning. Albumin level may be an indicator of the severity of hypoglycemia.[br][br]Nothing to Disclose: IB, SPK, SS, LES 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 967 300 2129 MON-208 PO11-01 Monday 1797 2012


1794 ENDO12L_MON-209 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Glycemic Control Trends in U.S. Hospitals Curtiss B Cook, Sophie Bersoux, Gail L Kongable, Jianfen Shu Mayo Clinic Arizona, Scottsdale, AZ; Mayo Clinic Arizona, Scottsdale, AZ; The Epsilon Group, Charlottesville, VA; University of Virginia, Charlottesville, VA Inpatient hyperglycemia is a high visibility topic, but little is known about trends in glucose control in US hospitals. We used the Remote Automated Laboratory System-Plus (Medical Automation Systems, Charlottesville, VA) to extract intensive care unit (ICU) and non-intensive care unit (non-ICU) inpatient point-of-care bedside glucose (POC-BG) data from 126 hospitals for the period January to December 2009 and compared it to previously reported glycemic control data analyzed from the same hospitals collected January to December 2007 (1). Hospital characteristics in 2009 were: 36% [lt]200 beds, 22% 200 to 299 beds, 13% 300 to 399 beds, and 29% [ge]400 beds; 63% were urban, 28% rural, 9% academic, 33% were in the South, 29% in the Midwest, 22% in the West, and 16% in the Northeast[bold]. [/bold]There were a total of 12,541,929 POC-BG measurements from 1,010,705 patients analyzed for 2007 and 10,659,418 measurements from 656,206 patients analyzed for 2009. Between 2007 and 2009, the proportion of patient-day weighted mean POC-BG values that were [gt]180 mg/dl among non-ICU patients declined from 28% to 25% (p [lt].001). Between the 2 study periods severe hypoglycemia ([lt]50 mg/dL) decreased in the non-ICU from 0.92% to 0.82% (p [lt].001), but no significant changes were noted for ICU patients. For non-ICU data, patient-day weighted mean POC-BG levels decreased significantly within each subgroup of hospital size, type, and region (all p [lt].001). The overall decrease of patient-day weighted mean POC-BG for non-ICU in 2009 is 5 mg/dL compared with 2007. For ICU patients the overall patient-day weighted mean POC-BG levels increased slightly but significantly (1 mg/dL, p [lt].001). The ICU subgroups that experienced the most increase in glucose levels were hospitals with [ge]400 beds (increased 2 mg/dL), academic institutions (increased 5 mg/dL) and hospitals in the West (increased 5 mg/dL) (all p [lt]0.001). In this first analysis of glucose trends in a large sample of US hospitals, we conclude that glycemic control is improving in the non-ICU population, with little improvement observed in the ICU patients. Hospitals appear to be effectively achieving improvements in glucose control in non-ICU patients while actually decreasing prevalence of severe hypoglycemia. Ongoing analysis will determine if these trends continue, and study is required to determine how hospitals are achieving better glycemic control in the non-critically ill so that protocols can be standardized and disseminated.[br][br]1. Cook CB, Kongable GL, Potter DJ, Abad VJ, Leija DE, Anderson M. Inpatient glucose control: a glycemic survey of 126 U.S. hospitals. Journal of Hospital Medicine 2009 4: E7-E14.[br][br]Disclosures: CBC: Coinvestigator, The Epsilon Group. GLK: Employee, The Epsilon Group. JS: Consultant, The Epsilon Group. Nothing to Disclose: SB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 30 300 2130 MON-209 PO11-01 Monday 1798 2012


1795 ENDO12L_MON-210 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Rigorous Delineation of Normal Glucose Levels during Hospitalization Jonathan Ownby, Lawrence Phillips, Qi Long, Wenqiong Xue, Christine Jasien, Darin Olson, Anne Tomolo, Arun Mohan, Sandra Jackson, Nadine Dubowitz, Diana Barb, Mary Rhee, Phyllis Watson-Williams Emory University, Atlanta, GA Although mortality during hospitalization is increased by hyperglycemia and may be increased by aggressive treatment, there has been no rigorous identification of normal glucose levels during hospitalization. Since such information is needed to constitute a basis for assessment of risk and a target for management, we evaluated hospital glucose levels in 4,595 veterans from SC, GA, and AL who were hospitalized for at least 3 days, and had continuity of care for two years before and three years afterwards. The 1,173 patients without either a diagnosis of diabetes or use of a diabetes drug throughout this period were defined as normal (NL), and their glucose values were compared with those in 3,178 patients who had diabetes throughout (DIAB), 84 initially diagnosed with diabetes during hospitalization (HOSP DIAB), 160 diagnosed 1-3 yr afterwards (1-3YR DIAB), and random plasma glucose levels in 977 healthy adult volunteers who had normal glucose levels during an OGTT (NGT OUTPT). The hospital patients had mean age 64 yr, and were 97% male, 57% white, and 37% black; 6% had unknown race. Mean (95% CI) glucose values for each group were NL 115 (73-186), DIAB 175 (56-411), HOSP DIAB 180 (75-500), 1-3YR DIAB 143 (70-260), and NGT OUTPT 93 (73-124). The glucose values in the NL group were significantly different from those in the other groups (all p[lt]0.0001), using either t tests (with only the highest, lowest, and median values from each patient, to limit bias from differences in numbers of samples), or linear mixed models (using all glucose values). NL patients had median glucose 104, 75th percentile 127, and 95th percentile 186 mg/dl, all higher than in NGT OUTPTs, who had median glucose 91, 75th percentile 99, and 95th percentile 124 mg/dl [ndash] glucose elevation presumably from the stress of illness and hospitalization. Among the 84 HOSP DIAB patients, only 60% had one glucose value above 186 mg/dl (95th percentile of the NL group), and only 40% had two glucose values above 186 mg/dl [ndash] possibly indicating inappropriate diagnoses. Conclusion: During hospitalizations, patients who most likely do not have diabetes exhibit glucose levels which are higher than in healthy normal glucose tolerant outpatients. The 95th percentile of [ldquo]normal[rdquo] hospital glucose is 186 mg/dl; a diagnosis of diabetes could be considered in patients who have glucose values above this level, and as a first objective, management could be aimed at keeping glucose values below this level.[br][br]Sources of Research Support: Veterans Administration Health Services Research and Development award IIR 07-138 awarded to LSP.[br][br]Nothing to Disclose: JO, LP, QL, WX, CJ, DO, AT, AM, SJ, ND, DB, MR, PW-W 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1322 300 2131 MON-210 PO11-01 Monday 1799 2012


1796 ENDO12L_MON-211 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Glycemic Control in the Intensive Care Unit: Lower Targets and Risk of Hypoglycemia, the Westchester Medical Center Experience Maria I Davila, Mallika Bhat, Guy Valiquette New York Medical College, Valhalla, NY Hyperglycemia in critically ill patients has been associated with increased mortality and complications. In 2004 Goldberg [italic]et al[/italic].(1) obtained tight glycemic control without significant hypoglycemia using an insulin protocol with target blood glucose of 100-139mg/dl. In 2009 the NICE-SUGAR study(2) showed that intensive glucose control increased mortality with significant increase in severe hypoglycemic events. Since NICE-SUGAR, a target of 140-180mg/dl has become the standard of care.[br]This study assesses glucose control in Westchester Medical Center Intensive Care Units (ICU) and the safety of GlucoCare System. We hypothesize that a target of 100-139mg/dl does not cause significant hypoglycemia. The GlucoCare System is a computerized system used to assist the nursing staff to adjust intravenous (IV) insulin therapy according to the 2004 Yale insulin protocol.[br]Methods: We obtained records from the GlucoCare system of all patients started on IV insulin in the ICU from July to December 2010. We reviewed charts retrospectively to assess the patient population, insulin requirements, hypoglycemic events, comorbidities, complications and APACHE II score.[br]Results: We reviewed 114 charts. The average patient was 69 years of age, 68% male, 32% known diabetics, admitted for cardiothoracic surgery. The average blood glucose level while on GlucoCare was 138.7mg/dL (107-176mg/dL). There were a total of 20 episodes of hypoglycemia of which 10 were on the same 5 patients. Of the hypoglycemic episodes, 12 ranged from 61-70mg/dL, 5 ranged from 51-60mg/dL and only 3 episodes ranged from 40-50mg/dL. There were no hypoglycemic episodes below 47 mg/dL. All hypoglycemic events were treated with 12.5-25g glucose IV and resolved without sequelae. 50% of the patients were discharged home, 42% to a rehabilitation center and there were 7 deaths, of which 4 had hypoglycemic events during their hospital course, none of which were at time of death. The average APACHE II score was 9.38 (2-21) and in those who died, was from 10-21.[br]Conclusion: Glucose control in the intensive care unit is crucial to improve patient outcome, morbidity and mortality. Our study had no hypoglycemic events less than 40mg/dl, contrary to the NICE SUGAR study who had 6.8% incidence of severe hypoglycemia in the intensive treatment group. We conclude that a glucose target range of 100-139mg/dl can be safely achieved with the use of a computerized system protocol, like GlucoCare.[br][br]1 - Goldberg PA et al., Diabetes Care 2004; 27:461. 2 - The NICE-SUGAR Study Investigators., N Engl J Med 2009; 360:1283.[br][br]Nothing to Disclose: MID, MB, GV 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1234 300 2132 MON-211 PO11-01 Monday 1800 2012


1797 ENDO12L_MON-212 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) LOGIC-Insulin Algorithm-Guided Versus Nurse-Directed Blood Glucose Control during Critical Illness: The LOGIC-1 Single-Center Randomized Controlled Clinical Trial Tom Van Herpe, Dieter Mesotten, Pieter J Wouters, Jeroen Herbots, Evy Voets, Jo Buyens, Bart De Moor, Greet Van den Berghe KU Leuven, Leuven, Belgium; KU Leuven, Leuven - Heverlee, Belgium Background: Tight blood glucose control in critically ill patients is difficult and labor intensive, resulting in poor glycemic control efficacy and an increased hypoglycemia rate. The LOGIC-Insulin computerized algorithm has been developed to assist nurses in titrating insulin to maintain blood glucose levels between 80-110 mg/dL and to avoid severe hypoglycemia ([lt]40 mg/dL).[br]Methods: In the LOGIC-1 randomized, single-center trial, 300 patients (heterogeneous mix of post-cardiac surgery, other surgical and medical ICU admissions) were randomized, by concealed computer allocation, to nurse-directed glycemic control (Nurse-C) or to algorithm-guided blood glucose control (LOGIC-C).[br]Results: The baseline characteristics of 151 Nurse-C patients and 149 LOGIC-C patients did not differ (age: 62[plusmn]14 y vs 65[plusmn]15 y, 61.6% vs 59.1% male, 21.2% vs 21.5% diabetes, APACHE-II: 21 (IQR 14-33) vs 19 (IQR 14-32)). Study time was 1.9 (IQR 1.1-3.7) days for Nurse-C and 1.9 (IQR 1.2-4.7) days for LOGIC-C (P=0.42). The mean blood glucose levels did not differ between the 2 groups (Nurse-C: 106[plusmn]11 mg/dL, LOGIC-C: 106[plusmn]9 mg/dL, P=0.36). The Glycemic Penalty Index, a marker of efficacy of blood glucose control, decreased from 12.3 (IQR 8.2-18.5) in Nurse-C to 9.8 (IQR 6.0-14.5) in LOGIC-C (P[lt]0.0001). The HyperGlycemic Index was also lower in LOGIC-C (2.5 (IQR 1.2-4.4) mg/dL), compared with Nurse-C (4.2 (IQR 1.5-7.3) mg/dL) (P=0.0028). The proportion of study time spent in the target range of 80-110 mg/dL was higher for LOGIC-C (68.5[plusmn]16.7%) than for Nurse-C patients (60.1[plusmn]18.8%) (P=0.00016). The mean daily difference between the min and the max blood glucose level, a marker of glycemic variability, was lower in LOGIC-C (31 (IQR 24-45) mg/dL) than in Nurse-C (37 (IQR 27-46) mg/dL) (P=0.045). The proportion of hypoglycemic measurements [lt]40 mg/dL was decreased (Nurse-C 0.13%, LOGIC-C 0%, P=0.015), but not when considered as a proportion of patients (Nurse-C 3.3%, LOGIC-C 0%, P=0.060). However, the interval between blood glucose measurements, a marker of workload, was slightly shorter in LOGIC-C (2.2[plusmn]0.4 h), compared with Nurse-C (2.5[plusmn]0.5 h) (P[lt]0.0001).[br]Conclusions: The computerized algorithm LOGIC-Insulin improved the efficacy of blood glucose control without increasing the rate of hypoglycemia in comparison with blood glucose control by the expert Leuven nursing team.[br][br]Sources of Research Support: FWO Senior Clinical Investigator (DM), IWT-TBM-100793, IOF-HB/10/039 and FWO-G.0557.08; LOGIC-1 ClinicalTrials.gov number, NCT01420302[br][br]Nothing to Disclose: TVH, DM, PJW, JH, EV, JB, BDM, GVdB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 945 300 2133 MON-212 PO11-01 Monday 1801 2012


1798 ENDO12L_MON-213 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Management of Glucocorticoid-Induced Hyperglycemia (GIH) in Hospitalized Patients with Acute Exacerbation of COPD (AECOPD) Shadi Abdelnour, Diana Engineer, Chuck Baimbridge, Elena Ruocco, Ron Giveon, Prasad Manian, Glenn R Cunningham University of Nevada School of Medicine, Las Vegas, NV; Baylor College of Medicine, Houston, TX; St Luke[apos]s Episcopal Hospital, Houston, TX [bold]Introduction[/bold]: Glucocorticoids (GC[apos]s) decrease insulin sensitivity and increase gluconeogenesis, which can cause hyperglycemia. GC[apos]s frequently are prescribed for patients with AECOPD and other medical conditions; however, there are no randomized trials or evidence-based guidelines for managing GIH. We are conducting a randomized prospective pilot study with two protocols, GL (Glargine/Lispro) and GLN (Glargine/Lispro/NPH). [bold]Objectives[/bold]: [underline]Primary[/underline]: To compare average daily glucose (ADG) readings on days 2-5 of the two protocols. [underline]Secondary[/underline]: To compare the incidence of hyperglycemia ([gt]180 mg/dL) and severe hypoglycemia ([lt]50 mg/dL) with each protocol. [bold]Methods[/bold]: In both arms, patients who are hyperglycemic [fasting glucose (FG) [gt]125 mg/dL or random glucose (RG) [gt]200 mg/dL] or known to have diabetes are started on lispro 0.2 units/kg/day divided between 3 meals and glargine 0.2 units/kg/day. The glargine dose is increased daily by 10% and 20% if FG levels are 141-200 mg/dL and [gt]200 mg/dL, respectively. The hyperglycemic effects of GC[apos]s initially are treated with 0.1 unit of insulin/kg/10 mg of prednisone or its equivalent with a maximum of 0.4 units of insulin/kg per 40 mg of prednisone or its equivalent. Patients receive either additional lispro in the GL protocol or NPH in GLN protocols. Lispro and NPH doses in the GL and GLN arms are increased daily by 10% and 20% if the preprandial glucose readings are 141-200 mg/dL and [gt]200 mg/dL, respectively. The target FG and RG levels are 90-140 mg/dL and [lt]180 mg/dL, respectively. Point of care glucose levels are obtained before each meal, at bedtime and at 2 AM. [bold]Results[/bold]: Nineteen patients have been enrolled. The ADG level was 172.7[plusmn]51.4 mg/dL and 150.2[plusmn]48 mg/dL for day 2 and 5, respectively in the GL arm, and 156.1[plusmn]45.8 mg/dL and 136.5[plusmn]34.2 mg/dL in the GLN arm. There were 1/232 (n/total n) and 0/178 of glucose values [lt]50 mg/dL in GL and GLN arms, respectively. There were 88/232 (38%) and 52/178 (29%) glucose values [gt]180 mg/dL in GL and GLN arms, respectively. No significant differences between the two protocols in ADG, change in ADG between Day 2 and 5, or in percent of glucose levels [lt]50 mg/dL or [gt]180 mg/dL have been noted. [bold]Conclusion[/bold]: Initiation and adjustment of insulin doses based on weight, dose of GC and glucose level seem to provide gradual improvement in ADG readings between day 1 and day 5, and to be a relatively safe and efficient means for improving glucose control.[br][br]Disclosures: GRC: Advisory Group Member, Abbott Laboratories, Endo Pharmaceuticals Inc., GlaxoSmithKline; Speaker, Abbott Laboratories, Merck & Co. Nothing to Disclose: SA, DE, CB, ER, RG, PM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 286 300 2134 MON-213 PO11-01 Monday 1802 2012


1799 ENDO12L_MON-215 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Fasting and Postprandial Glucose Metabolism on Basal-Bolus Versus Conventional Basal Insulin Therapy in Insulin-Na[iuml]ve Type 2 Diabetes Subjects Ravikumar Balasubramanian, Jean Gerrard, Roy Taylor Newcastle University, Newcastle-upon-Tyne, UK Objective: To test the hypothesis that an intensive basal bolus insulin regimen (mealtime insulin lispro + NPH insulin at bedtime) will be superior to a conventional insulin regimen (NPH insulin twice daily) in suppressing postprandial glucose excursions in insulin-na[iuml]ve adults with poorly controlled type 2 diabetes.[br]Research Design and Methods: 7 subjects with poorly controlled type 2 diabetes (age 30-70 years; 5M/2F; [ge] 2 years duration of diabetes; no history of weight loss or ketonuria at diagnosis; HbA1c [gt]7.5 %) on maximal oral hypoglycaemic therapy were recruited. After a 4-week run-in period, postprandial metabolism was studied using a standard liquid mixed meal test. All oral hypoglycaemic therapy was then stopped and each subject randomized to a 12-weeks of either (i) a conventional insulin regimen (CIR) with NPH insulin twice daily or (ii) a basal bolus regimen (BBR): meal-time insulin lispro + NPH basal insulin at bedtime. After 12 weeks, subjects crossed over to the alternate insulin regimen for a further 12 weeks. Individual dose adjustments were made in all subjects during both treatment periods by standardized weekly visits. Repeat postprandial metabolism study days were conducted at the end of each period of insulin therapy.[br]Results: Mean baseline HbA[sub]1c [/sub]was high pre-treatment (9.8 [plusmn] 1.1 %) and decreased significantly following both insulin regimes, but did not differ between the two treatment periods (CIR: 7.8 [plusmn] 0.7 % vs. BBR: 7.8 [plusmn] 0.8 % p=NS). Mean fasting glucose at baseline was 11.7 [plusmn] 1.0 mmol/L and this decreased significantly following both treatment periods, with a greater decrease following CIR compared to the BBR (8.0 [plusmn] 0.9 vs. 10.2 [plusmn] 0.9 mmol/L; p= 0.014). However, mean peak postprandial glucose (14.3 [plusmn] 1.0 vs. 14.6 [plusmn] 1.3 mmol/L, CIR and BBR respectively, p=NS) as well as mean postprandial glucose AUC (4444 [plusmn] 515 vs. 4815 [plusmn] 486 mmol/L*6h, CIR and BBR respectively, p=NS) were similar following both treatment arms.[br]Conclusions: Conventional and intensive basal bolus regimens were equally effective in improving metabolic control in insulin-na[iuml]ve type 2 diabetes subjects. Fasting glucose levels were slightly better controlled on the conventional regimen and contrary to our proposed hypothesis, postprandial glucose metabolism did not differ between the two groups. In type 2 diabetes, pre-meal insulinisation is an important determinant of post-prandial glucose control.[br][br]Sources of Research Support: Lilly UK Research Grant to RT.[br][br]Disclosures: RT: Research Funding, Eli Lilly & Company. Nothing to Disclose: RB, JG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 454 300 2135 MON-215 PO11-01 Monday 1804 2012


1800 ENDO12L_MON-216 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Safety and Efficacy of a Peri-Operative Protocol for Patients with Diabetes Treated with Continuous Subcutaneous Insulin Infusion Marilyn Augustine, Sandra Sobel, Melissa Ditmore, Amy C Donihi, David Miller, Jodie Reider, Patrick Forte, Mary Korytkowski University of Pittsburgh Medical Center, Pittsburgh, PA; University of Pittsburgh Medical Center, Pittsburgh, PA; University of Pittsburgh School of Pharmacy, Pittsburgh, PA; University of Pittsburgh Medical Center, Pittsburgh, PA; University of Pittsburgh School of Medicine, Pittsburgh, PA As the use of continuous subcutaneous insulin infusion (CSII) via insulin pumps for glycemic management in patients with type 1 and type 2 diabetes (DM1 and DM2) becomes more frequent, the number of CSII treated patients who are admitted for surgical procedures also rises. Currently there is no standardized approach for peri-operative glycemic management (PGM) of this group of patients. To address this, a PGM protocol (PGMP) was developed and implemented at our institution. The primary goal of this project was to investigate the safety and efficacy of a standardized PGMP for patients who use CSII when they are admitted for a surgical procedure.[br]All patients identified as using CSII at time of admission to the Same Day Surgery Unit between June and December 2011 were included in this analysis. Twenty-one patients using CSII underwent 28 surgeries during the 29 week period. These patients had a mean age of 51.0 [plusmn] 11.8 years and mean BMI of 32.8 [plusmn] 7.0 kg/m[sup]2[/sup]; 60.7% were female. Their mean duration of diabetes was 27.6 [plusmn] 12.4 years, with mean A1C 7.4 [plusmn] 1.0%. Mean basal insulin dose was 34.0 [plusmn] 17.8 units per day. Fourteen surgeries were on individuals with DM1, 11 on those with DM2, one with transplant related DM, and 2 with unknown type. Surgeries included 7 GI, 4 thoracic, 4 vascular, 3 neurological, 3 ENT, 3 orthopedic, 2 urological, and 2 dermatological procedures.[br]Patients were continued on CSII in 20 procedures (71.4%) and converted to IV insulin in 6 (21.4%). One patient did not receive any insulin during surgery, and one received IV insulin in addition to CSII. Patients who were continued on only CSII had shorter surgical duration (77.1 [plusmn] 55.6 vs. 253.8 [plusmn] 137.8 minutes), less likelihood of general anesthesia (55.0% vs. 87.5%) and lower pre- and post-operative blood glucoses (176.7 [plusmn] 61.2 and 162.7 [plusmn] 45.2 vs. 216.7 [plusmn] 67.0 and 208.4 [plusmn] 69.9 mg/dl) compared to patients whose surgical glycemic management was not CSII alone.[br]Six patients had their route of insulin delivery changed in the PACU. There were no CSII malfunctions noted peri-operatively. One patient experienced hypoglycemia preoperatively with no occurrence of intra- or post-operative hypoglycemia. Continuation of CSII did not change decision to admit for any patient.[br]A PGMP for patients using CSII is both safe and effective and helps reduce variability in clinical management of these patients.[br][br]Disclosures: MK: Principal Investigator, Sanofi-Aventis. Nothing to Disclose: MA, SS, MD, ACD, DM, JR, PF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 653 300 2136 MON-216 PO11-01 Monday 1805 2012


1801 ENDO12L_MON-217 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) To Evaluate the Safety of Insulin Glargine in Pregnancy Sameer Ansar, Sadia Mian, Matthew Hebdon, Ved V Gossain Michigan State University, East Lansing, MI Aim:[br]Although Insulin Glargine (IG) is being used in pregnancy, it has not been approved by the FDA and the data on its safety is limited. IG has been labeled as pregnancy category C. We performed this study to evaluate the safety of IG in pregnant women with diabetes.[br]Methods:[br]We reviewed the charts of women with gestational diabetes, type 1 diabetes and type 2 diabetes who were treated with IG before and during pregnancy. The recorded data included maternal and fetal morbidity and mortality.[br]Results:[br]From the maternal and fetal center at Sparrow Hospital (a 733-bed tertiary care hospital in Lansing, Michigan), 92 women were identified during the period of 2000-2009 who were treated with IG. 32 (35%) had gestational diabetes, 13 (14%) had T1DM, and 47 (51%) had T2DM. 18 women were on IG before pregnancy and 74 were started on IG during pregnancy. The mean HbA1c was 7.7 [plusmn] 2.37, 7.1 [plusmn] 1.88, and 6.3 [plusmn] 1.38 in first, second and third trimester. 31% of the woman had hypoglycemic episodes of various severities. No maternal deaths were reported.[br]One pregnancy resulted in intrauterine fetal death. 22% of the patients had preterm deliveries. Rate of caesarean section were 45% and mean gestational age was 36.2 [plusmn] 4.38 weeks. 12 % of the patients had macrosomia (defined as birth weight [gt]4000 grams), 2 % had shoulder dystocia, and 7% had neonatal hypoglycemia.[br]Discussion:[br]The data from our study compares favorably to the outcome from prior studies with pregnant patients on NPH insulin where intrauterine deaths were 1.7%, mean age at gestation was 37[plusmn]0.7, and fetal macrosomia was present in 15% of newborns. 7 % of the neonates had shoulder dystocia and 19% had hypoglycemia (1).[br]Since we completed this study others have reported the outcomes of pregnancies in women treated with IG. There has been only one prospective study (Negrato et al) comparing IG with NPH insulin in patients with gestational and pregestational diabetes.(2). They reported a decrease in adverse maternal and neonatal outcome with IG compared with NPH treated patients.[br]A review article by Kevin M Pantolone et al in Endocrine Practice looked at 23 reports 1001 pregnancies found no identifiable, consistent, adverse maternal or fetal outcome with the use of IG(3).[br]Conclusion:[br]Insulin Glargine in pregnancy appears to be safe during pregnancy in terms of maternal and fetal morbidity and mortality. The outcome of this study compares favorably with outcomes of prior studies using older insulins such as NPH insulin.[br][br](1)Pollex E, Moretti M, The annals of Pharmacology 2001;45(1)L1-8. (2) Negrato CA, Rafacho A, Negrato G, et al. Glargine vs NPH insulin therapy in pregnancies complicated by diabetes: an observational cohort study. Diabetes Res Clin Pract 2010;89:46[ndash]51. (3) Kevin M. Pantalone, Charles Faiman, and Leann Olansky MD. Endocrine Practice Vol No.3 may/June 2011.[br][br]Nothing to Disclose: SA, SM, MH, VVG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2224 300 2137 MON-217 PO11-01 Monday 1806 2012


1802 ENDO12L_MON-218 POSTER SESSION: Diabetes Pharmacotherapy (1:30 PM-3:30 PM) Safety and Efficacy of Linagliptin in Type 2 Diabetes (T2D) Patients with Common Renal and Cardiovascular Risk Factors Maximilian von Eynatten, Yan Gong, Angela Emser, Hans-Juergen Woerle Boehringer Ingelheim, Ingelheim, Germany [bold]Background[/bold]: Vascular complications are the leading cause of morbidity and mortality in T2D. Hypertension and/or microalbuminuria predict renal and cardiovascular (CV) outcome. This large pooled analysis from a global development program evaluated the safety and efficacy of linagliptin in this particularly high-risk population.[br][bold]Methods[/bold]: Data from 6 randomized, double-blind, placebo (PBO) controlled, phase 3 trials were analyzed (N=3119). Patients with microalbuminuria (urine albumin-to-creatinine ratio [UACR] 30-300 mg/g) and hypertension (systolic BP [ge]140 and/or diastolic BP [ge]90 mmHg; and/or taking antihypertensive drug; and/or history of hypertension) at baseline were eligible for this analysis. Patients treated with monotherapy (18-24 wks, depending on study design) received linagliptin 5 mg once daily or PBO. Patients treated with combinations of agents received linagliptin as initial combination with pioglitazone (PIO), 24 wks; add-on to metformin (MET), 24 wks; add-on to SU, 18 wks; or add-on to MET+SU, 24 wks. Efficacy endpoints were change from baseline in HbA1c and fasting plasma glucose (FPG) at 18 or 24 wks. Descriptive analysis of safety included incidence and severity of adverse events (AE), organ-specific AE, hypoglycemia episodes, and CV events.[br][bold]Results[/bold]: Of 3119 patients, 512 had both microalbuminuria and hypertension per screening criteria: 366 for linagliptin and 146 for PBO. Baseline demographics and characteristics were similar between groups: mean [SD] age, 59.5 [10.1] yrs; BMI, 29.9 [5.1]; HbA1c, 8.3 [0.9]%; and FPG, 176.9 [48.6] mg/dL. Proportions of patients with eGFR (MDRD) [ge]90, 60-[lt]90, and 30-[lt]60 mL/min were 51.8%, 40.6%, and 7.6%. Linagliptin significantly lowered HbA1c: -0.57% at 18 wks and -0.65% at 24 wks (vs 0% and -0.05% for PBO); [italic]P[/italic][lt]0.0001 at each time. Adjusted mean (SE) change in FPG from baseline (linagliptin-placebo) was -10.6 (3.9) mg/dL at 18 wks ([italic]P[/italic]=0.0074) and -21.3 (4.9) mg/dL at 24 wks ([italic]P[/italic][lt]0.0001). The AE and serious AE rates were similar for linagliptin vs PBO (AE 62.6% vs 62.3%; serious AE 4.1% vs 6.2%). Hypoglycemia increased in trials with SU: 19.8% for linagliptin vs PBO 5.9%, but was [lt]1% for linagliptin as monotherapy or add-on to MET or PIO.[br][bold]Conclusion[/bold]: In T2D patients with hypertension and microalbuminuria, linagliptin was well tolerated and achieved significant improvements in glycemic control. Linagliptin may support long-term preventive strategies to reduce risk of CV events and declining renal function.[br][br]Sources of Research Support: Boehringer Ingelheim.[br][br]Disclosures: MvE: Employee, Boehringer Ingelheim. YG: Employee, Boehringer Ingelheim. AE: Employee, Boehringer Ingelheim. H-JW: Employee, Boehringer Ingelheim. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1416 300 2138 MON-218 PO11-01 Monday 1807 2012


1803 ENDO12L_MON-225 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Epidemiology of Adverse Reaction to Statins in Routine Care Settings Huabing Zhang, Jorge Plutzky, Stephen Skentzos, Fritha Morrison, Perry Mar, Maria Shubina, Alexander Turchin Peking Union Medical College Hospital, Chinese Academy of Medical Sciences [amp] Peking Union Medical College, Beijing, China; Brigham and Women[apos]s Hospital, Boston, MA; Brigham and Women[apos]s Hospital, Boston, MA; Partners HealthCare System, Boston, MA; Partners HealthCare System, Boston, MA Background[br]Statins prevent cardiovascular events and their discontinuation increases mortality. In clinical trials, the frequency of adverse reactions is similar in statin and placebo groups. However, in routine care statins are commonly stopped due to adverse reactions. Systematic data on adverse reactions to statins in routine practice are limited.[br]Design[br]We conducted a retrospective cohort study of adverse reactions to statins at outpatient practices affiliated with two academic hospitals. All adult patients who received a statin prescription between 01/01/2000 and 12/31/2008 were included. Information on statin adverse reactions was obtained from a combination of structured EMR entries and analysis of electronic provider notes by validated natural language processing software. The first reported adverse reaction to a statin was studied for every patient.[br]Results[br]Among 104,957 patients who were prescribed statins during the study period, adverse reactions were documented for 18,763 (17.9%). More than a quarter (27.2%) of adverse reactions were myalgias or myopathy; other musculoskeletal and connective tissue symptoms were the second most common category (14.4%).[br]Over half (11,068/59.0%) of the patients who had an adverse reaction had the statin discontinued at least temporarily. Of these, 6,502 (58.7%) patients were re-challenged with a statin over the subsequent 12 months. The majority of patients who were re-challenged (92.1%) were still taking a statin 12 months after the adverse reaction. Among the 2,136 patients who were re-challenged with the same statin to which they had an adverse reaction, 1,511 (70.7%) were on the same statin 12 months later, 980 of them on the same or higher dose.[br]After adjustment for confounders, patients who had a mild ([ge] 3-fold) CK or LFT elevation were at a higher risk both for statin discontinuation and not being re-challenged after discontinuation. However, among patients who were re-challenged, neither CK nor LFT elevation was predictive of long-term discontinuation.[br]Conclusions[br]In this large retrospective study adverse reactions to statins were common and often led to their discontinuation. However, the majority of patients who are re-challenged were able to take statins (even the same ones to which the original adverse reaction was ascribed) long-term. This suggests that many of the adverse reactions may have other etiologies, are tolerable or are specific to individual medications rather than to the entire class of statins.[br][br]Sources of Research Support: Grant from the National Library of Medicine (1RC1LM010460).[br][br]Nothing to Disclose: HZ, JP, SS, FM, PM, MS, AT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 636 301 2139 MON-225 PO08-02 Monday 1808 2012


1804 ENDO12L_MON-226 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Plasma PCSK9 Levels Are Related with Markers of Cholesterol Synthesis in Familial Combined Hyperlipidemia Martijn CGJ Brouwers, Robert J Konrad, Thomas M van Himbergen, Aaron Isaacs, Seiko Otokozawa, Jason S Troutt, Ernst J Schaefer, Marleen MJ van Greevenbroek, Anton FH Stalenhoef, Jacqueline de Graaf Maastricht University Medical Centre, Maastricht, Netherlands; Eli Lilly and Company, Indianapolis, IN; Human Nutrition Research Center on Aging at Tufts University, Boston, MA; Erasmus Medical Centre, Rotterdam, Netherlands; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands [bold]Objective: [/bold]Familial combined hyperlipidemia (FCHL) is the most prevalent, complex genetic dyslipidemia in western society. FCHL patients are characterized by elevated plasma apolipoprotein B and triglyceride levels against a background of (abdominal) obesity and insulin resistance (1). Two recent independent studies showed that patients with FCHL have elevated plasma levels of proprotein convertase subtilisin kexin type 9 (PCSK9), an important regulator of LDL receptor expression, and markers of cholesterol synthesis (2,3). Both PCSK9 expression and cholesterol synthesis are downstream effects of hepatic activation of sterol regulatory element binding protein 2 (SREBP2). The present study was conducted to study the relationship between plasma PCSK9 and markers of cholesterol synthesis in FCHL.[br][bold]Methods: [/bold]Markers of cholesterol synthesis (squalene, desmosterol, lathosterol), cholesterol absorption (campesterol, sitosterol, cholestanol) and PCSK9 levels were measured in plasma of FCHL patients (n=103) and their normolipidemic relatives (NLR; n=240).[br][bold]Results: [/bold]Plasma PCSK9, lathosterol and desmosterol levels were higher in FCHL patients than their NLR (p[lt]0.001, age and sex adjusted). Heritability calculations demonstrated that approximately 35% of the variance in PCSK9 levels could be explained by additive genetic effects (p[lt]0.001). Significant age and sex adjusted correlations were observed for the relationship between PCSK9 and lathosterol, both unadjusted and adjusted for cholesterol, in the overall FCHL population (p[lt]0.001). None of the unadjusted cholesterol absorption markers were associated with PCSK9 levels. Multivariate regression analyses, with PCSK9 as the dependent variable, showed that the regression coefficient for FCHL status decreased by 25% (from 0.8 to 0.6) when lathosterol was included. Nevertheless, FCHL status remained an independent contributor to plasma PCSK9 levels (p[lt]0.001).[br][bold]Conclusion: [/bold]The present study confirms the previously reported high and heritable PCSK9 levels in FCHL patients. Furthermore, we now show that high PCSK9 levels are, in part, explained by plasma lathosterol, suggesting that SREBP2 activation partly accounts for elevated PCSK9 levels in FCHL.[br][br](1) Brouwers MC et al. Nat Rev Endocrinol 2012; in press. (2) Brouwers MC et al. Clin Sci 2011; 121: 397. (3) Van Himbergen TM et al. Arterioscler Thromb Vasc. Biol 2010; 30: 113.[br][br]Disclosures: RJK: Employee, Eli Lilly & Company. JST: Employee, Eli Lilly & Company. Nothing to Disclose: MCGJB, TMvH, AI, SO, EJS, MMJvG, AFHS, JdG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 698 301 2140 MON-226 PO08-02 Monday 1809 2012


1805 ENDO12L_MON-227 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Favorable Effect of Lycopene Consumption on HDL-C Serum Levels. A Randomized, Open-Label, Single Blind, Clinical Trial Daniel Cuevas-Ramos, Paloma Almeda-Valdes, Emma Chavez-Manzanera, Griselda Brito-Cordova, Oscar Perez-Mendez, Francisco J Gomez-Perez Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Mexico City, Mexico [bold]Introduction: [/bold]Atherosclerosis complicated with myocardial infarction is one of the main mortality causes in Mexico. The increment of high density lipoproteins cholesterol (HDL-C) levels reduces cardiovascular risk. Previous non-randomized studies (1) have related licopene consumption in tomatoes with increment of HDL-C. [bold]Methods: [/bold]This is a longitudinal, comparative, randomized, open-label, and single blind clinical trial. Initially, complete lipid profile was measured for screening interested individuals (n=432). Those with low HDL-c (men [lt]40 mg/dl, and women [lt]50 mg/dl) and normal triglycerides ([lt]150 mg/dl) that fulfilled the study[apos]s selection criteria started a run-in period of two weeks with isocaloric diet (n=55). After completion, participants were randomized to receive 300g of cucumber (control group) or uncooked tomato. Cucumber was selected because it doesn[apos]t contain licopene, and its preparation is similar. A total of 52 individuals completed the follow-up. Three patients were eliminated due to poor study compliance (n=2), and gastric intolerance to the tomato (n=1). Lipid profile was measured at baseline and at the end of the study. In addition, anthropometric measures, caloric consumption, physical activity, smocking and alcohol consumption, simple sugars, and omega 3 acids, were quantified at baseline and every week throughout the study. [bold]Results: [/bold]52 subjects (women=41, 78%) with a mean age of 42[plusmn]15.5 years and mean BMI of 27.6[plusmn]5 kg/m2 were included. Similar level of HDL-C were observed in both the control group (n=26, 36.8[plusmn]7.2 mg/dl) and the tomato group (n=26, 36.5[plusmn]7.5 mg/dl, p=0.90) at baseline. After one month, a significant increment in HDL-C levels was identified in tomato group vs the control group (41.6[plusmn]6.9 vs. 35.8[plusmn]7.3 mg/dl, p=0.008). The increment range was from 1 to a maximum of 12 mg/dl (mean of 5.0[plusmn]2.8 mg/dl). A lineal regression model adjusted for age, gender, waist to hip ratio, BMI, triglycerides, simple sugars, alcohol, physical activity and omega 3 with delta of HDL-C (final-basal) as dependent variable, showed an independent association of tomato consumption with the increment in HDL-C (beta=5.9, t=6.3, p[lt]0.0001). [bold]Conclusions: [/bold]Tomato consumption (300 g = 2 medium tomatoes per day, equivalent to 30mg of licopene, approximately) during one month produced a significant mean increment of 5 mg/dl in HDL-C.[br][br](1) Blum A et al., Clin Invest Med 2006;29:298.[br][br]Nothing to Disclose: DC-R, PA-V, EC-M, GB-C, OP-M, FJG-P 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 109 301 2141 MON-227 PO08-02 Monday 1810 2012


1806 ENDO12L_MON-228 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Novel Mechanism for Cardiovascular Risk Reduction Induced by Acarbose Andres Leon-Suarez, Victoria Valles, Liliana Munoz-Hernandez, Griselda Brito-Cordova, Francisco Javier Gomez-Perez Instituto Nacional de Ciencias M[eacute]dicas y Nutrici[oacute]n, Salvador Zubir[aacute]n, Mexico City, Mexico [bold]Introduction:[/bold] Although a decrease in postprandial hyperglycemia induced by acarbose has been postulated as a mechanism for CV risk reduction, there is no compelling evidence about it.[sup]1, 2[/sup] Since in patients with insulin resistance, an increased in formation of highly atherogenic oxidizable particles like small and dense LDL has been proven, we explored the effects of acarbose on oxidized LDL (ox-LDL) in patients with glucose intolerance.[sup].3[/sup][br][bold]Objective:[/bold] To evaluate the changes in postprandial lipemia and LDL oxidation with acarbose in untreated patients with glucose intolerance as possible mechanisms for cardiovascular risk reduction.[br][bold]Methods:[/bold] Twenty patients were included in the study (14 women, 6 men) and were randomly assigned into 2 treatment groups: acarbose (8 patients) or placebo (12 patients). To decrease intolerance, drug dosage was escalated (25mg weekly) to a maximum daily dose of 150mg. Patients followed an isocaloric diet with no modification of physical activity. Standard mixed meal loads were given before and after 3 months of treatment. Glucose, insulin, triglycerides, total cholesterol (TC) and free fatty acids (FFA) were measured at 0,5,10,20,30,60,120,180,240 and 300 min. by conventional methods. ApoB and ox-LDL were measured at 0 and 300 min. by nephelometry and ELISA assays, respectively. Delta refers to final-basal levels.[br][bold]Results:[/bold] The delta of AUC of glucose showed a higher reduction in patients with acarbose vs. placebo (-3103.75 vs. -1235, p=0.03). AUC TC (50451.25 vs 56376.25, p=0.05) was lower in the acarbose group. We also found a post-treatment reduction in FFA (119 vs. 140.6 mEq/L, p=0.02) in the acarbose group vs. placebo. In addition, the delta for ApoB (-1.25 mg/dl vs. 6.65, p=0.03) and ox-LDL (-7.5 vs 0.5 mU/L) showed a higher reduction with acarbose vs. control group.[br][bold]Conclusions:[/bold] We observed significant lower increases of glucose, TC, FFA, ApoB and ox-LDL in the acarbose group. These findings suggest that the reduction of cardiovascular risk by acarbose could be explained by a switch to chylomicrons (less atherogenic) over VLDL resulting in a decreased formation and oxidation of small and dense LDL particles (more atherogenic).[br][br](1) Chiasson JL et. al. JAMA. 2003: 290, 486-494. (2) S. D. Poppitt. Nutrition [amp] Food Science, 2005, 1, 23-34. (3) R.M. Krauss, P.W. Siri. Med Clin N Am. 2004: 88, 897-909.[br][br]Nothing to Disclose: AL-S, VV, LM-H, GB-C, FJG-P 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1013 301 2142 MON-228 PO08-02 Monday 1811 2012


1807 ENDO12L_MON-229 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Lipoprotein Subfractions by Ion Mobility in Lean and Obese Children: Correlations with Gender, Adiposity, Inflammation and Puberty Matthew R Benson, Jobayer Hossain, Michael Caulfield, Dawn F Shalhout, Ligeia Damaso, Samuel Gidding, Nelly Mauras Nemours Children[apos]s Clinic, Jacksonville, FL; University of Florida College of Medicine, Jacksonville, FL; Alfred I Dupont Hospital for Children, Wilmington, DE; Nichols Institute, San Juan Capistrano, CA; Alfred I Dupont Hospital for Children, Wilmington, DE [bold]Background:[/bold] Lipoproteins comprise a heterogenous group of particles that differ in size, density, lipid and apolipoprotein concentration[bold]. [/bold]Measurement of small, atherogenic lipid subfractions may provide an additional biomarker to predict risk of cardiovascular disease in adults but this is not well studied in children. Ion Mobility Analysis is a novel assay designed to more accurately characterize circulating lipid and lipoprotein subfractions.[br][bold]Objective:[/bold] To establish normative data for lipoprotein subfractions in healthy lean children and to compare their data with a group of obese children pre-selected with normal glucose, blood pressure and plasma cholesterol and triglyceride concentrations. Subjects were part of a bigger study on inflammatory markers (1).[br][bold]Subjects and Methods:[/bold] Fasting blood samples in 162 children, mean age 12 [plusmn] 3yrs (7.0 [ndash] 18.9 yrs), both prepubertal and pubertal, 62% males, 75 lean (BMI[bold]: [/bold]18.6 [plusmn] 6.6 kg/m[sup]2[/sup]), 87 obese (BMI[bold]:[/bold] 31.7 [plusmn] 5.4) were analyzed using an ion mobility assay (2). Correlation of lipoprotein subfractions with anthropometric and laboratory markers was performed. Principal component analysis was used to avoid using correlated variables.[br][bold]Results:[/bold] Normative data for subfractions of LDL, HDL, VLDL and apolipoproteins were obtained in healthy children. Lean children had higher HDL-L (76%), HDL-S (13%), and HDL-total (27%) and also LDL-VS (17%) as compared to obese (p [lt] 0.01) and lower LDL-M (-30%, p [lt] 0.01) and M+S (-21%, p = 0.02) as well as LDL-total (-13%, p =0.035). In both groups the LDL-component was higher in males than females (p [lt]0.003) and pubertal vs. prepubertal children (p [lt] 0.01). Prepubertal children had a higher HDL-component than pubertal ones (p [lt] 0.004). Adjusting for gender and pubertal status LDL-and HDL-components were significantly correlated (positively and negatively correspondingly) with obesity (p [lt] 0.004).[br][bold]Conclusions:[/bold] Despite normal triglycerides and cholesterol measured with standard assays, ion mobility analysis showed significant differences in lipid and apolipoprotein subfractions between lean and obese children, even those prepubertal. Long-term, prospective follow up may better characterize the predictability of lipid subfractions for future cardiovascular disease risk in children.[br][br](1) Mauras N, et al., J Clin ENdocrinol Metab 2010; 95:1060. (2) Caulfield MP, et al., Clin Chem 2008; 54:1307.[br][br]Sources of Research Support: Thrasher Research Fund. Quest Diagnostics Laboratories.[br][br]Disclosures: MC: Employee, Quest Diagnostics. DFS: Employee, Quest Diagnostics. NM: Principal Investigator, Quest Diagnostics. Nothing to Disclose: MRB, JH, LD, SG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 629 301 2143 MON-229 PO08-02 Monday 1812 2012


1808 ENDO12L_MON-230 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Low-Dose Weekly Chloroquine Is Associated with Favorable Effects on Lipid Metabolism without Significant Influence on Insulin Resistance Lawrence S Lee, Melvin KS Leow, Ying Xu, Yin Bun Cheung, Nicholas I Paton National University of Singapore, Singapore, Singapore; Tan Tock Seng Hospital, Singapore, Singapore; Singapore Clinical Research Institute, Singapore, Singapore Background: Chloroquine is an antimalarial and immunomodulating drug. Daily doses of chloroquine/hydroxychloroquine increased insulin sensitivity and improved lipid metabolism at high doses given daily. We investigated whether low dose chloroquine given weekly could also be effective for treating metabolic syndrome.[br]Methods: We recruited 1516 healthy volunteers for a randomized placebo-controlled clinical trial of chloroquine to prevent influenza (CHIP Study) (1). Half were administered chloroquine base 300mg (chloroquine phosphate 500mg) daily for 7 days, followed by 300mg weekly for 10 more weeks. The other half were on placebo. We obtained fasted plasma samples from a subset. Samples were assayed for total cholesterol (CHOL), High-Density Lipoprotein-cholesterol (HDL-C), triglycerides (TG), insulin and glucose. Low-Density Lipoprotein-cholesterol (LDL-C) was calculated using the Friedewald equation. Homeostatic model assessment insulin resistance (HOMA-IR) and beta-cell function (HOMA-B) were evaluated using the HOMA2 calculator.[br]Results: 270 volunteers with fasting samples were recruited, 134 on chloroquine. One hundred and eighty-two were male, mean age was 27.8 years and body mass index was 23. kg/m2. After 12 weeks, TG in the chloroquine group was 11% lower relative to placebo group (95% CI -20% to 0%, P = 0.047). The CHOL:HDL-C ratio was 7% lower (95% CI: -11% to -1%, P = 0.013). There was a trend towards lower (-5%) LDL-C (95% CI: -11% to +1%, P = 0.075). There were no significant differences in HDL-C, insulin, glucose and HOMA indices between chloroquine and placebo.[br]Discussion [amp] Conclusion: Low dose chloroquine given weekly had modest but significant effects on lowering TG and CHOL:HDL-C. This cheap, safe and convenient treatment should be investigated further for efficacy in ameliorating metabolic syndrome and mitigating cardiovascular risk.[br][br](1) Paton et al., Lancet Infectious Diseases 2011; 11(9):655.[br][br]Sources of Research Support: National Medical Research Council Singapore Grant NMRC/H1N1R/003/2009.[br][br]Nothing to Disclose: LSL, MKSL, YX, YBC, NIP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 84 301 2144 MON-230 PO08-02 Monday 1813 2012


1809 ENDO12L_MON-231 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Metabolic Impact of Chronic Cannabis Smoking Sara Sable, Ranganath Muniyappa, Marilyn Huestis, Ronald Ouwerkerk, Ahmed M Gharib, Mary Walter, Amber Courville, Andrea Mari, Sara Aney, Gail Hall, Kong Y Chen, Nora D Volkow, George Kunos, Monica C Skarulis NIDDK, National Institutes of Health, Bethesda, MD; NIDA, National Institutes of Health, Baltimore, MD; NIDDK, National Institutes of Health, Bethesda, MD; NIDDK, National Institutes of Health, Bethesda, MD; National Research Council, Padova, Italy; NIAAA, National Institutes of Health, Bethesda, MD; NIAAA, National Institutes of Health, Rockville, MD [bold]Background:[/bold] Endocannabinoid activation of peripheral cannabinoid (CB1) receptors disrupts energy balance, promotes lipogenesis and induces hepatic steatosis, insulin resistance, glucose intolerance, and dyslipidemia. [Delta]-9-Tetrahydrocannabinol the primary psychoactive component of [italic]Cannabis Sativa[/italic] activates peripheral and central CB1 receptors. Cannabis is the most common illicit drug in the US, however, little is known of the metabolic effects of chronic cannabis smoking in healthy adults.[br][bold]Objective:[/bold] To determine if chronic cannabis smoking is associated with hepatic steatosis, insulin resistance, reduced [beta]-cell function, or dyslipidemia in healthy individuals.[br][bold]Methods: [/bold]This case-control study included cannabis smokers (n=30; women=12, men=18; mean age (SD): 27[plusmn]8 yr) and control subjects (n=30) matched for age, gender, ethnicity, and BMI. Hepatic fat (HF) content was measured by MRI spectroscopy; adiposity by DEXA (% total body fat, TBF), and abdominal MRI (visceral fat, VF and total abdominal fat, TAF); Insulin sensitivity by the quantitative insulin sensitivity check index (QUICKI), oral glucose insulin sensitivity (OGIS) index, adipocyte insulin resistance index (AIRI), and free fatty acid (FFA) suppression during an oral glucose tolerance test (OGTT); [beta]-cell function by OGTT modeling; and energy intake and Healthy Eating Index (HEI) by Diet History Questionnaire.[br][bold]Results: [/bold]Self-reported cannabis use was significant [years of use (median, range): 9.5 (2-38) yr; [ldquo]joints[rdquo]/day: 6 (1.3-30)]. Energy intake was similar while HEI was lower in cannabis smokers. There were no significant group differences in HF, TBF, or VF. Cannabis smokers had lower plasma HDL cholesterol (49[plusmn]14 vs. 55[plusmn]13 mg/dl, p=0.02), but fasting levels of glucose, insulin, total cholesterol, LDL cholesterol, triglycerides, or free fatty acids were not significantly different between the groups. All surrogate measures of insulin sensitivity or [beta]-cell function did not differ between the groups. Fasting levels of ghrelin, leptin, adiponectin, PYY, or GLP-1 were not different between smokers and controls.[br][bold]Conclusions: [/bold]Chronic cannabis smoking was not associated with hepatic steatosis, visceral adiposity, insulin resistance, or metabolic dysfunction. Several factors may contribute to our findings including desensitization of CB1 receptors, the anti-inflammatory and other salutary effects of CB2 receptor stimulation, and the modulatory effects of other phytocannabinoids, such as cannabidiol.[br][br]Nothing to Disclose: SS, RM, MH, RO, AMG, MW, AC, AM, SA, GH, KYC, NDV, GK, MCS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1724 301 2145 MON-231 PO08-02 Monday 1814 2012


1810 ENDO12L_MON-232 POSTER SESSION: Lipids (1:30 PM-3:30 PM) IGFBP-2 Is Associated with a Longitudinal Decline in Triglycerides in Type 2 Diabetes Ram Prakash Narayanan, Bo Fu, Anthony Payton, Simon G Anderson, Julie E Hudson, Robert L Oliver, Kirk W Siddals, John P New, Adrian Hugh Heald, William ER Olier, John M Gibson The University of Manchester, Salford, UK; The University of Manchester, Manchester, UK; The University of Manchester, Manchester, UK; The University of Manchester, Manchester, UK; The University of Manchester, Manchester, UK The insulin-like growth factor system is strongly implicated in diabetes and cardiovascular disease. IGF binding protein-2 (IGFBP-2) has a preferential affinity for IGF-II. Low concentrations of IGFBP-2 are associated with the metabolic syndrome and with elevated fasting triglyceride levels and hypertension, but the longitudinal effects of IGFBP-2 on serum triglycerides and blood pressure are not known. We studied the associations of circulating IGFBP-2 with longitudinal trends in triglycerides and blood pressure in type 2 diabetes.[br]Plasma IGFBP-2 measurements were performed on samples collected in 2002-03 in 489 individuals (294 males) with type 2 diabetes from Salford, Northwest England. We used a commercial ELISA (RayBio Inc) with intra-assay and inter-assay coefficients of variation (CV) of [lt]10% and [lt]12% respectively. Longitudinal clinical data were derived from a comprehensive integrated database of primary care and hospital records. Some serum triglyceride measurements may have been non-fasting. Linear mixed-effect regression analysis in STATA 10SE was used to model the relationship between baseline log-transformed IGFBP-2 concentration and longitudinal changes in serum triglycerides, systolic and diastolic blood pressure.[br]Mean age of the cohort was 62.9 years (95% CI 62.0-63.9). Mean IGFBP-2 concentration was 326.3 ng/ml (95% CI 308.3-344.2). Elevated baseline log-IGFBP-2 was associated with a decrease in serum triglycerides over an 8-year period ([beta] -0.35, [95% CI -0.54 to -0.15], p[lt]0.001).[br]Elevated log IGFBP-2 also predicted a longitudinal decrease in diastolic blood pressure ([beta] -1.14, [95% CI -2.2 to -0.07], p=0.037), but no effect was observed with respect to systolic blood pressure over the 8-year period.[br]The associations for IGFBP-2 were independent of gender, age, diabetes duration and concentrations of IGF-I, IGF-II, IGFBP-1 and IGFBP-3 and adjusted for the use of HMG CoA inhibitors and fibrates (for association with triglycerides) and with ACE inhibitors, diuretics and beta blockers (for systolic and diastolic blood pressure) over any duration within the study period.[br]This study demonstrates for the first time that circulating IGFBP-2 predicts a longitudinal decline of triglycerides and diastolic blood pressure in type 2 diabetes, and would suggest a persisting effect on metabolic risk factors. Therapeutic modulation of IGFBP-2 concentration may merit exploration as a means to decrease cardiovascular risk in individuals with type 2 diabetes.[br][br]Nothing to Disclose: RPN, BF, AP, SGA, JEH, RLO, KWS, JPN, AHH, WERO, JMG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1575 301 2146 MON-232 PO08-02 Monday 1815 2012


1811 ENDO12L_MON-233 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Generational Differences in Total and HDL Cholesterol and in (Weight*Height/HDL) of South Asian Immigrants Living in New Jersey Richard Ro, Era Caterina Murzaku, Mridula William, Silpa Gadiraju, Romona Kersellius, Sirisha Jonnalagadda, Asra Majeed, Deval Bhatt, Glen Monteiro, Louis Amorosa UMDNJ-Robert Wood Johnson Medical School, New Brunswick, NJ; UMDNJ-School of Public Health, Newark, NJ The South Asian population of New Jersey is third highest in the United States. Though premature coronary artery disease is clinically obvious in young South Asian male immigrants, cardiovascular (CV) risk factors have not been specifically defined by NHANES for this group. Health statistics of South Asians are generally included in the category of Asian and Pacific Islanders but this ethnic pool has marked variation in cardiovascular outcomes, which preclude analyses of a specific subgroup. We have had the opportunity to study metabolic factors that contribute to the CV risks in the South Asian population in New Jersey. We obtained lipid, glycemic, blood pressure (BP) and anthropometric measures in 436 male and female South Asians attending festive events. We categorized the data by age ([le]45 years versus [gt]45 years) and by sex. The average ages [all data are mean[plusmn] 2SEM(n)] of younger vs. older men were: 35 [plusmn]2(82) vs. 59[plusmn]2 (115) respectively. The comparable ages in women were 32[plusmn]2 (113) vs. 57[plusmn]2(114), respectively. Total and HDL cholesterol (mg/dl) for these age and sex groups were: 183[plusmn]7 and 37[plusmn]2 vs. 207[plusmn]7 and 44[plusmn]2 for men and 168[plusmn]6 and 49[plusmn]3 vs. 213[plusmn]6 and 53[plusmn]2 for women. Total cholesterol and HDL-C in men clearly differ from NHANES cholesterol values and trends observed in the general population. HDL-C in both sexes correlated with weight and height (-0.37 and -0.31, p [lt]0.01 for both). The data infer that body mass is a determinate of the HDL-C value, despite the fact that BMI did not differ between any of the sex or age categories.[br]We previously noted that the expression (weight/HDL-C) predicted increased therapeutic insulin requirements in patients immediately following coronary artery bypass graft surgery. (1) Post-operative CABG patients, including South Asians, are known to be markedly insulin resistant. We determined that a similar expression (weight*height/HDL-C) calculated from the data in our South Asian population correlated with systolic BP (0.12, p [lt]0.05) and hemoglobin A1c (0.15, p[lt]0.05). Systolic BP and A1c are commonly linked to insulin resistance. (Weight*height/HDL-C) sharply differentiated South Asians grouped by age and sex (F=48, one way ANOVA) better than any other measured metabolic variable. This simple mathematical expression in South Asians that encompasses components of metabolic risk might be a marker of risk for premature coronary artery disease.[br][br](1) William M et al., Abstracts AACE Annual Meeting, 2012.[br][br]Nothing to Disclose: RR, ECM, MW, SG, RK, SJ, AM, DB, GM, LA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 700 301 2147 MON-233 PO08-02 Monday 1816 2012


1812 ENDO12L_MON-234 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Hyperlipidemia Metabolic Control and Cardiac Events during Civil Unrest Saad Sakkal Metabolic Care Center, Aleppo, Syrian Arab Republic Objective: Civil unrest is stressful to all. Stress effect metabolic control in general, but no or rare studies showed the influence of civil unrest on Hyperlipidemia control and cardiac events in diabetes patients. We quantitated the effect during the recent Syrian civil unrest.[br]Methods: In our Diabetes clinic patients are seen quarterly. In each visit we review H[amp]P, laboratory parameters and undercurrents psychosocial factors.[br]In 200 patients we determined means difference in Cholesterol, TG, TG/HDL ratio and HgA1c as well as cardiac events(MI, CHF, revascularization or change in angina pattern)during 2 distinct comparable periods: for the 6 months before the civil unrest and after (July to December 2010, 2011).We excluded new onset diabetes patients and other new onset illnesses.[br]Results: In the period before civil unrest July to December 2010 There was a drop in Bp: 8mm systolic/7mm diastolic, FBS: 37mg/dl, 2hpp:62, HgA1c (2.5%) and insulin dose by 22% from baseline. At the same time there was a drop in Cholesterol: 22mg/dl (214 to 192), Triglycerides: 59(225 to 166), TG/HDL ratio: 1.31(5.11 to 3.8) implying less inflammation, and cardiac events by %50(6 to3). In the period during civil unrest July to December2011 there was a rise in Bp (26/8) FBS (47), 2hpp (128), HgA1C (2%) and insulin dose by 25%. At the same time there was an increase in Cholesterol: 17(188 to 205), TG: 115(152 to 267), TG/HDL ratio: 1.72(3.6 to 5.32) and cardiac events rate by %60(5 to 8).Expense increased from $3200 to 4800.[br]Conclusion: Diabetes Glucose, lipids, Bp, and cardiac control are affected by civil unrest, probably as a result of stress, lack of health resources or other factors. We need more studies for diabetes, Bp, Lipids, and cardiac status during unfortunate civil unrest events.[br][br]Nothing to Disclose: SS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 154 301 2148 MON-234 PO08-02 Monday 1817 2012


1813 ENDO12L_MON-235 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Statin-Related Rhabdomyolysis in a VA Patient Population Harshit R Shah, John C Jennings Loma Linda University, Loma Linda, CA; Loma Linda University - VA, Loma Linda, CA All statins (St) are reported to cause rhabdomyolysis (Rbm, ICD9: 728.88). The incidence varies with individual statins and is increased with certain combinations of St with other medications. The FDA has recently recommended restricting the use of high dose simvastatin (S) due to a substantially higher incidence of Rbm at the 80mg (compared with 20mg) dose of S in the SEARCH trial (1,2). Since that trial randomized patients to 2 doses of S irrespective of LDL-C and thus does not reflect the actual clinical use of S we determined the incidence of St associated Rbm in a VA population at high risk for CAD with multiple co-morbidities. We identified 102 cases of Rbm which required hospitalization in 100 patients between 7/1/2006-6/30/2011 of which 52 cases were associated with the use of lipid lowering agents. There were a total of 85534 patients with 427670 person years of St, fibrate, of combination use with the greatest use of S compared with the other St., Rbm (cases/10000 patient years (95% CI) occurred in association with each St as well as with fibrate monotherapy: S 0.68 (0.44-1.02), rosuvastatin 3.02 (0.98-7.07), lovastatin 1.21 (0.14-4.38), fluvastatin 11.73 (1.42-42.37), pravastatin 1.54 (0.03-8.6), gemfibrozil (G) 1.61 (0.19-5.84), and fenofibrate 0. Rbm occurred with each dose of S: 10mg 0.41 (0.04-1.48), 20mg 0.65 (0.28-1.28), 40mg 0.78 (0.35-1.49), and 80mg 0.78 (0.25-1.84). Compared with S monotherapy the incidence of Rbm increased with combination of S+G (600mg bid): 20mg 3.91 (0.47-14.15), 40mg 6.07 (1.25-17.76), and 18.16 (7.3-37.41).[br][underline][italic]Conclusions:[br][/italic][/underline]When used in a clinical setting where all statins were used and where dose was determined by titration based upon LDL-C there does not appear to be an increase incidence of Rbm with S compared with other statins or an apparent dose-response between 10 and 80 mg of S. On the other hand when S is used in combination with gemfibrozil there is a dose response increase in the incidence of Rbm. These results suggest that the restrictions on the use of S should be limited to combination therapy with S and G which should continue to be contraindicated for all dose combinations.[br][br]1) SEARCH study collaborative group. SEARCH trial Am Hart J 2007; 154:815-23, 823.e1-823.e6. 2) FDA : Limit use of 80mg Simvastatin. 6/8/2011.[br][br]Nothing to Disclose: HRS, JCJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 129 301 2149 MON-235 PO08-02 Monday 1818 2012


1814 ENDO12L_MON-236 POSTER SESSION: Lipids (1:30 PM-3:30 PM) The Relationship between Serum Circulating IGF-1 and Liver Fat in the United States Population Shauna S Runchey, Edward J Boyko, George N Ioannou, Kristina M Utzschneider University of Washington, Seattle, WA; VA Puget Sound Health Care System, Seattle, WA Serum insulin like growth factor-1 (IGF-1) is synthesized in the liver and increases insulin sensitivity when administered exogenously. Nonalcoholic fatty liver disease (NAFLD) is associated with insulin resistance and in two studies has been associated with lower IGF-1 levels. We aimed to determine whether serum IGF-1 levels are associated with the presence of NAFLD independently of potential confounders such as obesity and insulin resistance. We used data from the Third National Health and Nutrition Examination Survey (NHANES III; 1988-1994), selecting those subjects who had both a fasting blood draw and ultrasound examination (n=14,797). After excluding subjects with hepatitis C, significant alcohol intake, pregnancy or those taking glucose-lowering medications, data on 4571 adults (2540M/2031F, mean[plusmn]SE age 42[plusmn]0.5 y, BMI 26[plusmn]0.2 kg/m[sup]2[/sup]) were analyzed. Measures included fasting serum IGF-1, IGF binding protein-3 (IGFBP-3), glucose, insulin and Hgb A1C% as well as liver fat grade by ultrasound (normal, mild, moderate or severe). HOMA-IR was calculated as a measure of insulin resistance. Using bivariate linear models and chi-square analyses, we found that a higher level of IGF-1 was associated with female sex, younger age, race/ethnicity other than Mexican/Hispanic, lower BMI, lower waist circumference, lower HOMA-IR and lower Hgb A1C% (all P[lt]0.001). In a univariate logistic regression analysis, increasing serum IGF-1 level was associated with a lower risk of NAFLD (OR 0.75, 95% CI 0.66-0.84 for a 1 standard deviation increase in IGF-1). However, in multivariate logistic regression models, serum IGF-1 levels were not independently associated with presence of liver fat after adjustment for age, sex, race/ethnicity and BMI (OR 0.90, 95% CI 0.80-1.03) or waist circumference (OR 0.91, 95% CI 0.80-1.03). Adjustment for HOMA and Hgb A1C% did not significantly change the relationship between IGF-1 and liver fat. IGFBP-3 level was not associated with presence of liver fat in any model. Contrary to previous reports, we conclude that in a large population of adults, the association observed between IGF-1 levels and liver fat is confounded by adiposity and waist circumference. We further conclude that previous findings of a relationship between IGF-1 levels and liver fat cannot be explained by insulin resistance. IGF-1 is unlikely to play a role in the pathophysiology of NAFLD but instead appears as a correlate of conditions associated with NAFLD.[br][br]Sources of Research Support: Department of Veterans Affairs.[br][br]Nothing to Disclose: SSR, EJB, GNI, KMU 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1963 301 2150 MON-236 PO08-02 Monday 1819 2012


1815 ENDO12L_MON-237 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Predictors of Diastolic Dysfunction in Patients with Metabolic Syndrome Meghan Businaro, Mateusz Jasielec, Rami Doukky, Nicole Woodrick, Marcin Wiesyk, Lynne T Braun, Annabelle Volgman, Lynda H Powell, Rasa Kazlauskaite Rush University Medical Center, Chicago, IL; Rush University Medical Center, Chicago, IL; Rush University Medical Center, Chicago, IL Metabolic syndrome (MetS), diastolic dysfunction and diastolic heart failure are linked (1-3). Since there is no specific therapy for diastolic heart failure(4,5), it is important to identify risk factors related to diastolic dysfunction in patients with MetS for prevention of diastolic heart failure.[br]In a cross-sectional study, we recruited patients with MetS from clinics, affiliated with a university hospital. Patients with a history of symptomatic heart disease or diabetes were excluded. All patients underwent echocardiographic evaluation, and measurements of waist girth, blood pressure, fasting glucose, and lipids. Categorical definition of diastolic dysfunction includes a combination of abnormal diastolic parameters (6). In order to determine whether metabolic biomarkers differentially affect diastolic parameters, we selected E/A and E/e[apos], and tissue Doppler parameters e[apos] and a[apos], indicators of myocardial relaxation. Multivariable linear and logistic regression models were utilized to determine the individual MetS biomarkers most predictive of cardiac diastolic parameters and epicardial fat. For each dependent variable, the final model selected was a model that minimized the Akaike information criterion out of a set of candidate models composed of all possible combinations of the blood pressure, waist girth, triglyceridemia, glycemia, and HDL-cholesterol, controlling for age.[br]Among 98 asymptomatic participants with MetS (mean age 52[plusmn]8 years, 75% women, 65% ethnic minority), 36% had grade 1 diastolic dysfunction. The only predictor of diastolic dysfunction was diastolic blood pressure (OR=2.07, p=0.01). The best model to predict E/A ratio (R2=0.36) included triglyceridemia (beta=-0.09, p=0.02), HDL-c (beta=-0.05, p=0.10), and diastolic blood pressure (beta=-0.07, p[lt]0.01). Predictor of E/e[apos] (R2=0.15) was systolic blood pressure (beta=0.06, p[lt]0.05). Predictor of e[apos] (R2=0.36) was diastolic blood pressure (beta=-0.67, p[lt]0.01). Predictor of a[apos] (R2=0.08) was triglyceridemia (beta=0.49, p=0.02). We concluded that diastolic dysfunction may affect one-third of patients with MetS. Individual metabolic biomarkers (hypertension and dyslipidemia) are associated with impairment of diastolic parameters. It is imperative to perform a longitudinal study, exploring the effects of metabolic and lifestyle factors on the of diastolic parameters in MetS.[br][br]1. de las Fuentes L, et al. European Heart Journal 2007;28:553-9. 2. Aijaz B,et al. Mayo Clinic Proceedings 2008;83:1350-7. 3. Lam CSP, et al. Circulation 2011;124:24-30. 4. Owan TE, et al. N Engl J Med 2006;355:251-9. 5. de Groote P, et al. Arch Cardiovasc Dis 2008;101:361-72. 6. Paulus WJ, et al. European Heart Journal 2007;28:2539-50.[br][br]Sources of Research Support: William G. McGowan Charitable Fund.[br][br]Nothing to Disclose: MB, MJ, RD, NW, MW, LTB, AV, LHP, RK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2364 301 2151 MON-237 PO08-02 Monday 1820 2012


1816 ENDO12L_MON-238 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Non-HDL-Cholesterol/HDL-Cholesterol Ratio Is Better Than Apolipoprotein B/Apolipoprotein A1 Ratio To Identify Metabolic Syndrome in Korean Population Se Won Kim, Yoon Young Cho, Sun-Mi Park, Hye Jeong Kim, Ji Cheol Bae, Sun Wook Kim, Jae Hoon Chung, Jae Hyeon Kim Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea [bold]Objective[/bold][br]The ratio of apolipoprotein B (apoB) to apolipoprotein A1 (apoA1) has been reported to be associated with the metabolic syndrome (MetS) and insulin resistance. Non-HDL-cholesterol (non-HDL-C) is regarded as a surrogate marker for apoB in routine clinical practice. However, it is unclear whether the ratio of non-HDL-C to HDL-cholesterol (HDL-C) can replace apoB/apoA1 ratio to identify MetS. We aimed to compare the predictive value for the MetS between apoB/apoA1 ratio and non-HDL-C/HDL-C ratio.[br][bold]Methods[/bold][br]We performed a retrospective study of 48703 Korean adults (mean age, 51.0 years) who participated in a routine health screening examination. Anthropometry, blood pressure, fasting glucose, fasting insulin, CRP, lipid profiles, apoB and apoA1 were measured.[br][bold]Results[/bold][br]MetS, as defined by a modification of NCEP-ATP III criteria, was present in 24.6% of men and 12.0% of women. Among the components of the MetS, HDL-C showed the strongest correlation with apoB/apoA1 ratio ([italic]r [/italic]= -0.326, [italic]P [/italic][lt]0.001 in men, and [italic]r [/italic]= -0.595, [italic]P [/italic][lt]0.001 in women) and non-HDL-C/HDL-C ratio ([italic]r [/italic]= -0.714, [italic]P[/italic] [lt]0.001 in men, and[italic] r [/italic]= -0.697, [italic]P[/italic] [lt]0.001 in women). The odds for MetS were significantly higher in the highest compared with the lowest apoB/A1 ratio quartiles, after adjustment for confounding variables, in both men (odds ratio [OR]= 3.14, 95% CI=2.86-3.44) and women (OR= 8.17, 95% CI=6.71-9.95). As a function of the quartile of non-HDL-C/HDL-C levels, the ORs for the MetS were 9.23 (95% CI=8.26-10.32, [italic]P[/italic] [lt]0.001) in men and 19.56 (95% CI=15.16-25.23, [italic]P[/italic] [lt]0.001) in women, in the highest compared with the lowest quartiles. To compare the predictive value for MetS between these two lipid ratios, we analyzed the ROC curves of apoB/apoA1 ratio and non-HDL-C/HDL-C ratio. ROC analysis showed that the AUCs of non-HDL-C/HDL-C ratio (0.76 [95% CI, 0.75-0.77] in men and 0.84 [95% CI, 0.83-0.85] in women) were significantly higher than those of apoB/apoA1 ratio (0.66 [95% CI, 0.65-0.67] in men and 0.76 [95% CI, 0.75-0.77] in women). The associations of insulin and HOMA-IR were significantly stronger with non-HDL-C/HDL-C ratio than those with apoB/apoA1 ratio in both men and women.[br][bold]Conclusion[/bold][br]Our findings indicate that non-HDL-C/HDL-C ratio is a better marker than apoB/apoA1 ratio for identifying the MetS among Koreans.[br][br]Nothing to Disclose: SWK, YYC, S-MP, HJK, JCB, SWK, JHC, JHK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1364 301 2152 MON-238 PO08-02 Monday 1821 2012


1817 ENDO12L_MON-239 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Evaluation of Markers of Vascular Inflammation, Obesity and Insulin Resistance in Obese Children Reem Kheetan, Yoram Elitsur, Ronald Stanek, Abid Yaqub, Nalini Santanam Marshall University, Huntington, WV; Marshall University, Huntington, WV Objective:[br]The objective of our research was to study the levels of oxidized LDL(Ox-LDL), a marker of vascular inflammation in obese children and to evaluate its relationship with other markers of obesity and insulin resistance.[br]Materials and Methods:[br]In a cross sectional study, 28 obese and 16 non-obese children between the ages 8 and 18 yrs were prospectively recruited. Morphological measurements including height, weight and waist circumference was obtained and plasma levels of Oxidized LDL and Retinol Binding Protein (RBP-4, a marker for insulin resistance and obesity) were measured from all participants. BMI was calculated according to the CDC standards. Obesity was defined as having a body mass index (BMI) above 95th percentile for age and sex. Serum lipids, A1c, insulin and glucose levels were measured in obese children. HOMA-IR and HOMA-IS was calculated in obese children.[br]Results:[br]Oxidized LDL levels were found to be significantly higher in obese versus non-obese children (p[lt]0.001). Oxidized LDL correlated significantly in a positive manner with children[apos]s waist circumference (p=0.03), BMI (p=0.001), BMI percentile (p[lt]0.001) and BMI SDS (p[lt]0.001) scores. There was positive correlation found between Oxidized LDL and RBP-4 (p=0.02). There was no significant correlation found between Oxidized LDL and serum lipids, glucose, insulin, A1c, HOMA-IR and HOMA-IS in obese children group.[br]Conclusion:[br]Ox-LDL, a biomarker of vascular inflammation was found to be elevated in obese children as compared to non-obese children in our study population. Ox-LDL was positively correlated with various indices of obesity and with RBP-4. This provides evidence for obesity pre-disposing to vascular inflammation at an earlier age in an individual life.[br][br]1. Barlow SE, Dietz WH, 1998 Obesity evaluation and treatment: Expert Committee recommendations. The Maternal and Child Health Bureau, Health Resources and Services Administration and the Department of Health and Human Services. Pediatrics 102: E29. 2. Weiss R, Dziura J, Burgert TS, et al, 2004 Obesity and the metabolic syndrome in children and adolescents. N Engl J Med 350: 2362-2374. 3. U.S. Preventive Services Task Force, 2006 Screening and interventions for overweight in children and adolescents: recommendation statement. Am Fam Physician 73: 115-119.[Soft Break]4. Ogden CL, 2004 Defining overweight in children using growth charts. Md Med 5: 19-21. 5. Bundak R, Furman A, Gunoz H, et al, 2006 Body mass index references for Turkish children. Acta Paediatr 95: 194-198.[br][br]Nothing to Disclose: RK, YE, RS, AY, NS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1858 301 2153 MON-239 PO08-02 Monday 1822 2012


1818 ENDO12L_MON-240 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Magnetic Resonance Spectroscopy Quantification of NAFLD in Obese Adolescents on Vitamin D Therapy Carisse M Orsi, Jane L Lynch, Daniel E Hale University of Texas Health Science Center at San Antonio, San Antonio, TX [bold]Background and Aims:[/bold] Pediatric obesity is associated with non-alcoholic fatty liver disease (NAFLD), Type 2 diabetes and hypertension. Low serum 25-hydroxyvitamin D3 [25(OH)D] may contribute the risk for NAFLD and insulin resistance. This prospective longitudinal pilot study evaluates the relationship of 25(OH)D and fatty liver using Magnetic Resonance Spectroscopy (MRS) repeat measurements to quantify liver fat following Vitamin D replacement and normalization over six months.[br][bold]Materials and Methods:[/bold] We studied 9 obese (body mass index [gt]95%), Hispanic adolescents between the ages of 10 and 15 years with a diagnosis of NAFLD ([gt]5% fat) measured by MRS and 25(OH)D insufficiency. Other causes of fatty liver were excluded by measuring the following labs: Hepatitis B and C profile, anti-nuclear antibody, anti-LKM antibody, anti-smooth muscle antibody, alpha-1 antitrypsin level, ceruloplasmin, iron, and total iron binding capacity in each patient. All subjects were given a minimum of 2000 IU of Vitamin D for a period of 6 months and doses were increased as needed based on serum levels of 25(OH)D at monthly visits. Measurements of fatty liver, 25(OH)D, insulin, fasting plasma glucose, lipid profiles, and iPTH were measured pre- and post- treatment as well as anthropomorphic measurements of body mass index (BMI), waist-hip ratio were evaluated at monthly intervals.[br]Results: 12 subjects were evaluated and there were 3 screen fails. Each screen fail was due to having [lt]5% fat on MRS. All 9 subjects completed the study and maintained stable BMI (pre: 36.8 kg/m2 vs. post: 37.6kg/m2, P=0.29) and plasma glucose (pre: 88.9 mg/dL vs. post: 89.1mg/dL P=0.72) over the 6-month trial. 25(OH)D levels improved with required dosages ranging from 2000-8000 IU per day (pre: 18.9 ng/dl vs. post: 39.2 ng/dl, P[lt]0.01). MRS baseline levels were significantly elevated at 31.2% and decreased to 23.1% (P=0.07) over 6 months while the alanine aminotransferase (ALT) levels slightly trended upwards (pre: 66.7 U/L vs. post: 75.8 U/L, P=0.69).[br][bold]Conclusions:[/bold] Normalization of serum 25(OH)D levels in obese, Hispanic adolescents led to decreased MRS liver fat while maintaining BMI in this pilot study. No significant effect was observed in lipid profiles, blood pressure, waist-to-hip ratios and fasting insulin. The change in NAFLD was not consistent with serum ALT measurements. Further studies are needed using MRS measures to provide accurate measures of liver fat.[br][br]Sources of Research Support: Lester and Liesel Baker Foundation Award; Institute for Integration of Medicine and Science Clinical and Translational Science Award at the UTHSCSA; Christus Santa Rosa Pediatric Children[apos]s Hospital; CHART center.[br][br]Nothing to Disclose: CMO, JLL, DEH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 845 301 2154 MON-240 PO08-02 Monday 1823 2012


1819 ENDO12L_MON-241 POSTER SESSION: Lipids (1:30 PM-3:30 PM) A High Normal TSH Level Is Associated with an Atherogenic Lipid Profile in Euthyroid Non-Smokers with Newly Diagnosed Asymptomatic Coronary Heart Disease Wanjia Xing, Chenggang Wang, Aihong Wang, Xiaomei Yang, Jiajun Zhao, Chunxiao Yu, Jin Xu, Yinglong Hou, Ling Gao Provincial Hospital Affiliated to Shandong University, Jinan, China; Shandong University of Traditional Chinese Medicine, Jina, China; Provincial Hospital Affiliated to Shandong University, Jinan, China; Qianfoshan Hospital, Jinan, China; Provincial Hospital Affiliated to Shandong University, Jina, China Introduction: A patient[apos]s lipid profile may be influenced by thyroid function, but the detailed mechanism remains unclear. Increasing evidence suggests that TSH may exert extra-thyroidal effects. The goal of this study was to evaluate the relationship between serum TSH levels and the lipid profile in euthyroid non-smokers with newly diagnosed asymptomatic coronary heart disease (CHD).Methods: A retrospective study was conducted including 406 euthyroid non-smokers (187 males and 219 females, age ranged 45-88 years) newly diagnosed with asymptomatic CHD from 2004 to 2010 in Jinan, China.Lipid parameters and the levels of TSH, thyroid hormone were determined. Patients were divided into low-normal TSH subgroup (G1, TSH 0.3-0.99 mIU/L), a moderate- normal TSH subgroup (G2, TSH 1.0-1.89 mIU/L) and a high-normal TSH subgroup (G3, TSH 1.9-4.8 mIU/L). Regression analysis and odds ratios (ORs) were used to assess the influence of parameters on the lipid profile and to estimate the risks of dyslipidemia.Results: In these study subjects, the ORs of TSH with respect to the risk of hypercholesterolemia and hypertriglyceridemia were 1.640 (P=0.002) and 1.349(P=0.017), respectively. The TC level increased by approximately 0.42 mmol/L in G2 and by 0.43 mmol/L in G3 (P[lt]0.01). The serum level of LDL-C was approximately 1.11 times higher in the G2 subgroup than in the G1 subgroup (P[lt]0.05). The prevalences of hypercholesterolemia in G2 and G3 were 4 times than that in G1 (P[lt]0.01 in both). For hypertriglyceridemia, the prevalence was approximately 2-folds greater in G2 and G3 than that in G1 (P[lt]0.05 and P[lt]0.01, respectively). The TSH level, even within the normal range, was positively linearly correlated with total cholesterol(TC), non-high density lipoprotein cholesterol (non-HDL-C) and triglycerides (TG) (B=0.017, 0.026 and 0.027, respectively, P[lt]0.01).With each 1mIU/L increase in the level of TSH, the levels of TC, TG and non-HDL-C will increase by 1.010, 1.064, and 1.062mmol/L, respectively: TSH levels were correlated in a linear positive manner with the TC, non-HDL-C and TG levels in euthyroid non-smokers with newly diagnosed asymptomatic CHD. TSH might exert adverse effects on the lipid profile and might represent a risk factor for hypercholesterolemia and hypertriglyceridemia in the context of CHD.Further studies are needed with a much greater and well-matched number of individuals to estimate the real impact of this association and its clinical significance.[br][br]Sources of Research Support: This research is supported by National Natural Science Foundation of China(81170794, 30971409), Natural Science Foundation (ZR2009CZ009) and the international cooperation grant (2011) of Shandong Province of china.[br][br]Nothing to Disclose: WX, CW, AW, XY, JZ, CY, JX, YH, LG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 908 301 2155 MON-241 PO08-02 Monday 1824 2012


1820 ENDO12L_MON-242 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Preliminary Observations about Statin Therapy in Heart Transplant Recipients with Muscular Dystrophy Ekta Singh, Sudhir Kushwaha, Walter K Kremers, Yogish C Kudva Mayo Clinic, Rochester, MN; Mayo Clinic, Rochester, MN; Mayo Clinic, Rochester, MN [bold]Background[/bold]: Statin use is standard practice after heart transplantation (HT) to prevent/delay cardiac allograft vasculopathy. HT recipients with muscular dystrophy (MD) represent a special cohort, since statins are generally avoided in myopathies.[br][bold]Methods: [/bold]We reviewed the HT database at Mayo Clinic Rochester, MN from January 1996 through December 2011, and studied the medical record for documentation of muscle-related complications from statin use in HT recipients with MD.[br][bold]Results: [/bold]4 patients (all males) with MD underwent HT between Jan.1996 and Dec. 2011. Two (50%) patients had Becker[apos]s MD, one had Emery Dreifuss MD, and one had an unclassifiable, poorly defined MD. The median age at the time of HT was 29.5 years (25-36). All patients had minimal myopathic symptoms, and were fully functional prior to HT.[br]All patients received pravastatin post-HT with the dose titrated to achieve an LDL-C goal of [le] 100 mg/dL. The mean dose was 45 mg (20-80). One patient was changed to Simvastatin 40 mg due to suboptimal LDL-C control. The patients tolerated pravastatin well, without any functional impairment for a median duration of 54 months (13-96). The median LDL-C during therapy was 84 mg/dL (70-119).[br]One patient developed progressive weakness 7 years after HT, and 1 year after changing treatment to simvastatin, prompting statin discontinuation, that was followed by mild improvement in symptoms. One patient experienced myalgias 2 years after HT, after pravastatin dose was increased to 40 mg. One patient reported myalgias on pravastatin 40 mg 8 years after HT, prompting its discontinuation that did not result in any symptomatic improvement. One patient reported no adverse events 1 yr after HT, on pravastatin 20 mg. There were no serious adverse effects, including rhabdomyolysis. EMG was not performed on any patient. Creatine kinase levels fluctuated widely, and did not correlate with patients[apos] muscular symptoms.[br]With the exception of 1 case of mild subclinical hypothyroidism, all patients had normal thyroid function. Three patients were on Sirolimus, and one on Tacrolimus-based immunosuppression. There was no reported use of medications interfering with statin metabolism.[br][bold]Conclusion: [/bold]Long-term, medium-dose pravastatin therapy is well tolerated by patients with MD undergoing HT. Creatine kinase is not a reliable predictor of adverse muscle-related events. Since these patients are rare, a multi-center effort is required to study them.[br][br]Nothing to Disclose: ES, SK, WKK, YCK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1884 301 2156 MON-242 PO08-02 Monday 1825 2012


1821 ENDO12L_MON-243 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Metabolic Effects of Walnuts in Patients with Prediabetic Metabolic Syndrome Mustafa Kemal Balci, Bengi Balci, Pinar Hoda Mediterranean University, Antalya, Turkey; Mediterranean University, Antalya, Turkey Background:[br]The relationship between dietary contents and Metabolic Syndrome (MetS) are well-known. With a well structured diet and lifestyle changes, MetS prevelance can be significantly decreased.[br]Objectives:[br]In our study, we aimed to demonstrate consumption of 10 gr walnuts per day for 3 months in patients with prediabetic MetS diagnosed according to NCEP ATP III criterias.[br]Methods:[br]The present study includes 60 newly diagnosed patients with prediabetic MetS defined by NCEP ATP III criterias, who are 33 females and 27 males. Participants are randomly separated to two groups, as 30 in healthy nutrition (HN) group and 30 in specified healthy nutrition (SHN) group. The only diet difference between HN and SHN group was 10 gr walnuts intake per day in SHN group. Individuals were evaluated for body weight, waist circumference, blood pressure and lipid profiles (Total cholesterol, LDL cholesterol, HDL cholesterol and triglyseride levels) at the beginning of the trial and after the 3 months period.[br]Results:[br]In HN group; at the beginning of the trial and after the 3 months period, Body weight; 88,9[plusmn]16.1 and 76,3[plusmn]13.1 kg (p[lt]0.001), waist circumference; 108[plusmn]13.8 and 95.8[plusmn]10.3 (p[lt]0.001), systolic blood pressure; 146[plusmn]12 and 142[plusmn]8 mmHg (p[lt]0.002), diastolic blood presure; 84[plusmn]8 and 79 [plusmn]7(p[lt]0.003) have been measured. There weren[apos]t any changes in Total cholesterol, HDL cholesterol, Triglyseride and LDL cholesterol levels.[br]In SHN group; Body weight 90.8[plusmn]12 and 79.1[plusmn]10.9 kg (p[lt]0.0001), waist circumference 109.5[plusmn]11.8 and 97.4[plusmn]9.9 cm (p[lt]0.0001), Systolic blood pressure 147[plusmn]14 ve140[plusmn]9 (p[lt]0.0001), diastolic blood pressure 85[plusmn]7 and 79[plusmn]6 mmHg, triglyseride 278[plusmn]231 and 210 [plusmn]141 mg/dl(p[lt]0.021), HDL cholesterol 36[plusmn]9 and 43[plusmn]14 mg/dl (p[lt]0.004) have been measured.[br]Conclusion:[br]Overall, in SHN group; triglyseride levels decreased and HDL cholesterol levels increased after the 3 months. We need further studies to determine the effects of walnuts in long term.[br][br]Nothing to Disclose: MKB, BB, PH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1482 301 2157 MON-243 PO08-02 Monday 1826 2012


1822 ENDO12L_MON-244 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Association between Fibroblastic Growth Factor 21 and Selenoprotein P and Nonalcoholic Fatty Liver Disease According to Obesity Status Ji A Seo, Chai Ryoung Eun, Hyunjoo Cho, Ho Cheol Hong, Hae Yoon Choi, Yoon Jung Kim, Joo Hyung Kim, Sae Jeong Yang, Hye Jin Yoo, Hee Young Kim, Sin Gon Kim, Kyung Mook Choi, Sei Hyun Baik, Dong Seop Choi, Chol Shin, Nan Hee Kim Korea University Medical School, Seoul, Republic of Korea; Korea University Medical School, Seoul, Republic of Korea [bold]Background[/bold]: The role of hepatokines, such as fibroblastic growth factor (FGF) 21 and selenoprotein P (SeP), in metabolic diseases has been mostly studied in obese population. This study aimed to investigate whether the associations between nonalcoholic fatty liver disease (NAFLD) and the circulating levels of FGF21 and SeP were different according to the body mass index (BMI) in Korean adults.[br][bold]Methods[/bold]: Among 3,264 participants of the Korean Genome Epidemiology Study, 80 subjects with normal weight (BMI[lt]23 kg/m[sup]2[/sup]) were matched for age and sex to 240 overweight/obese subjects (BMI [ge]23 kg/m[sup]2[/sup]). They were further classified according to the presence/absence of NAFLD within each group. Subjects with diabetes, history of viral hepatitis or alcohol drinking [ge]140 g/wk were excluded. NAFLD was diagnosed by computed tomography using the liver attenuation index (LAI), the difference between mean hepatic and splenic attenuation score [LAI [lt]5 HU (NAFLD), LAI [ge]5 HU (no-NAFLD)]. Serum FGF21 and SeP were measured using the commercial human ELISA kits.[br][bold]Results[/bold]: Serum FGF21 and SeP levels were positively correlated with BMI [r=0.11 (p=0.043) and r=0.22 (p=[lt]0.001)], waist circumference [r=0.15 (p=0.007) and r=0.29 (p=[lt]0.001)], diastolic blood pressure [r=0.17 (p=0.002) and r=0.16 (p=0.004)], triglyceride [r=0.27 (p=[lt]0.001) and r=0.13 (p=0.023)], and homeostasis model assessment (HOMA-IR) [r=0.14 (p=0.011) and r=0.14 (p=0.014)]. Only in normal weight group, FGF21 and SeP levels were higher in subjects with NAFLD than in those without after adjusting for age and sex [mean(se); log of FGF21, 5.29(0.09) vs. 5.64(0.09), p=0.008, log of SeP 1.44(0.14) vs. 2.10(0.14), p=0.001 in no-NAFLD vs NAFLD, respectively]. The odds ratios (ORs) of FGF21 and SeP for the presence of NAFLD were increased in normal weight group [OR(95% CI), FGF21 4.76(1.62-14.01); SeP 2.51(1.36-4.65)] after adjusting for age, sex, BMI, triglyceride, alanine transaminase (ALT) and HOMA-IR. But this association was not seen in overweight/obese group [OR(95% CI), FGF21 1.67(0.99-2.81); SeP 1.22(0.86-1.72)].[br][bold]Conclusion[/bold]: Increased serum FGF21 and SeP levels were highly associated with NAFLD even after adjusting for the major risk factors of NAFLD. This association was evident only in normal weight Koreans.[br][br]Nothing to Disclose: JAS, CRE, HC, HCH, HYC, YJK, JHK, SJY, HJY, HYK, SGK, KMC, SHB, DC, CS , NHK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 803 301 2158 MON-244 PO08-02 Monday 1827 2012


1823 ENDO12L_MON-245 POSTER SESSION: Lipids (1:30 PM-3:30 PM) The Characteristics of the Subjects with Very Low Level of LDL Cholesterol and the Risk of Intracerebral Hemorrhage Sang Ah Lee, Gwanpyo Koh, Dae Ho Lee Jeju National University School of Medicine and Jeju National University Hospital, Jeju, Korea Hypocholesterolemia are found frequently, but the clinical implication of the hypocholesterolemia has not been studies a lot. Some studies revealed an association between hypocholesterolemia and intracerebral hemorrhage (ICH). We aimed to evaluate the clinical characteristics of the subjects with very low serum low density lipoprotein cholesterol (LDL-C) and compare the risk of ICH by various clinical parameters. From our hospital records, we evaluated the clinical characteristics of subjects with LDL-C 40 mg/dL or less (very low LDL-C group). Then, we evaluated the risk of ICH in this very low LDL-C group and in subjects with low LDL-C group ([lt] 70 mg/dL). Among a total of 34415 subjects who were presented to the laboratory to measure serum LDL-C, 250 subjects had serum LDL-C 40 mg/dL or less that we defined as very low level. Nearly half of the subjects had very low LDL-C level without history of statin use, usually due to alcohol intake or various chronic illnesses including liver diseases, end stage renal disease (ESRD). ICH occurred in 3 subjects with very low LDL-C, all of whom had no history of statin use. Among various clinical factors, ESRD tended to be related with ICH in subjects with serum LDL-C less than 70 mg/dl. In conclusions, we found about 1% of subjects whose LDL-C was measured in a hospital have LDL-C 40 mg/dL or less. Among them, half of the subjects had no history of hypolipidemic therapy. ICH incidence was not related with LDL-C level or statin use.[br][br]Nothing to Disclose: SAL, GK, DHL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1200 301 2159 MON-245 PO08-02 Monday 1828 2012


1824 ENDO12L_MON-246 POSTER SESSION: Lipids (1:30 PM-3:30 PM) Metabolic Effects of Metreleptin Treatment in Familial Partial Lipodystrophy (FPL) Elif A Oral, Adam Neidert, Bashir Hakim, Stephan Miller, Kevin Shan, Boman Irani, Karen Lutz, Jean L Chan University of Michigan, Ann Arbor, MI; Amylin Pharmaceuticals, Inc, San Diego, CA Previous studies have shown that patients with lipodystrophy (LD) and leptin deficiency significantly improve glycemic control and lipid levels when treated with metreleptin (recombinant-methionyl human leptin, an investigational drug). However, low leptin levels ([lt]7-12 ng/mL in earlier studies) are not uniformly observed, especially in patients with partial LD. Here we report data on the effect of metreleptin treatment in a cohort of FPL patients who were diagnosed clinically, without using a leptin threshold as an eligibility criterion.[br]Nineteen FPL patients with diabetes and/or high triglyceride (TG) levels (18F/1M, age 46[plusmn]12y) were enrolled in an ongoing open-label expanded access protocol (sponsored by Amylin Pharmaceuticals, Inc.) at University of Michigan between Mar 2009 and Oct 2011. Baseline (BL) A1C was 7.9[plusmn]1.6%, TG level 264[plusmn]188 mg/dL, and leptin level 14.6[plusmn]11.2 ng/mL (mean[plusmn]SD, range 1.4-43 ng/mL). As of the data cut-off, 11 and 8 patients had completed 6 and 12 mo study visits, respectively. Patients received an average daily dose of 7.3[plusmn]2.0 mg metreleptin/day by QD or BID SC injection. Concomitant diabetes medications were kept constant or adjusted down to avoid hypoglycemia.[br]Overall, A1C and TG improved with metreleptin treatment. Eight of 19 patients had both elevated A1C [ge]7% and TG [ge]200 mg/dL at BL while some patients presented with only one abnormality, necessitating subgroup analyses. In patients with A1C [ge]7% (14/19, mean 8.5[plusmn]1.3%), A1C decreased by -0.4[plusmn]0.6% at 6 mo (n=10) and 1.0[plusmn]0.7% at 12 mo (n=7, range 4.7, +0.4%). In those with TG [ge]200 mg/dL (12/19, mean 348[plusmn]188 mg/dL), TG decreased by -31[plusmn]28 mg/dL at 6 mo (n=6) and -184[plusmn]127 mg/dL at 12 mo (n=5, range -644, +129 mg/dL). Metabolic benefits were observed across a range of BL leptin levels (1.4-23 ng/mL). While on metreleptin, 2 of 12 patients on oral antidiabetic agents (OAD) were able to discontinue an OAD, and 4 of 9 patients on insulin reduced their insulin dose by 20% or more.[br]Metreleptin was generally well tolerated. Nausea (26%) and injection site reactions (47%) were the most commonly reported adverse events related to treatment. No serious adverse events were assessed as related to treatment. Metreleptin treatment in this cohort of FPL patients diagnosed clinically without selection for low leptin levels resulted in clinically meaningful improvements in A1C and TG, suggesting that measuring leptin levels is not needed prior to initiating metreleptin treatment in LD patients.[br][br]Sources of Research Support: Amylin Pharmaceuticals, Inc.[br][br]Disclosures: EAO: Investigator, Amylin Pharmaceuticals; Consultant, Amylin Pharmaceuticals. SM: Employee, Amylin Pharmaceuticals; Stockholder, Amylin Pharmaceuticals. KS: Employee, Amylin Pharmaceuticals; Stockholder, Amylin Pharmaceuticals. BI: Employee, Amylin Pharmaceuticals; Stockholder, Amylin Pharmaceuticals. KL: Employee, Amylin Pharmaceuticals; Stockholder, Amylin Pharmaceuticals. JLC: Employee, Amylin Pharmaceuticals; Stockholder, Amylin Pharmaceuticals. Nothing to Disclose: AN, BH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1752 301 2160 MON-246 PO08-02 Monday 1829 2012


1825 ENDO12L_MON-254 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Physiologic Elevation of Endogenous GH/IGF-I Levels in a Mouse Model Does Not Accelerate Carcinogen-Induced Mammary Gland Tumorigenesis Manuel D Gahete, Jose Cordoba-Chacon, Chike Anadumaka, Daniel D Lantvit, Raul M Luque, Steven M Swanson, Rhonda D Kineman University of Illinois at Chicago, Chicago, IL; University of Illinois at Chicago, Chicago, IL; University of Cordoba, Cordoba, Spain Several epidemiologic studies suggest that circulating levels of GH/IGF-I are positively correlated with breast cancer risk. These observations coupled with the fact that mouse models of developmental GH/IGF-I deficiency or resistance are less susceptible to breast cancer development, have led to the conclusion that GH/IGF-I axis plays a role in breast carcinogenesis. However, no studies to date have shown that high physiologic levels of circulating GH/IGF-I can promote mammary tumorigenesis. We have previously developed and characterized an unique genetically engineered mouse model (HiGH mice) with elevated endogenous GH/IGF-I output within the physiologic range due to a GH-cell specific knock-out of the IGF-I and insulin receptors. Consistent with the anabolic effects of GH/IGF-I axis, HiGH mice are slightly bigger than controls and exhibit elevated insulin levels and insulin resistance; however, reproductive function is intact and prolactin levels are comparable to controls. Of note, the relative elevation of plasma IGF-I found in HiGH mice (20-40%) is comparable to the 31% increase in circulating IGF-I levels shown to increase breast cancer risk in premenopausal women and, therefore, we have used the HiGH mice to test the hypothesis that elevated GH/IGF-I levels (either directly or indirectly through alterations in metabolic function) sensitize the mammary gland to chemical-induced mammary gland tumor formation. Our data support the positive role of the GH/IGF-I axis on mammary gland development, as 6-week-old HiGH females exhibit more developed mammary glands than age-matched controls. Since moderate elevation of plasma IGF-I has been shown to not affect mammary gland development, these data suggest that GH acting directly or indirectly through stimulation of local IGF-I production in the mammary gland is responsible for the observed phenotype. Strikingly, our data indicate that mammary tumor formation 25 weeks after carcinogen (DMBA) administration is not increased in mice with endogenous elevation of the GH/IGF-I axis. In fact, there is a non-significant reduction in tumor multiplicity and a significant delay in tumor latency. In addition, the number of mice that develop large tumors ([gt]1cm) was reduced in HiGH mice. Therefore, our results suggest that a developmental elevation in GH/IGF-I levels within the physiological range hastens mammary gland development but does not accelerate carcinogen-induced tumorigenesis in the mouse mammary gland.[br][br]Sources of Research Support: Fundacion Caja Madrid (M.D.G.); Fundacion Alfonso Martin Escudero (J.C.C); RYC-2007-00186, BFU2010-19300, CIBERObn (MICINN/FEDER), (R.L.M).[br][br]Nothing to Disclose: MDG, JC-C, CA, DDL, RML, SMS, RDK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 901 302 2161 MON-254 PO43-01 Monday 1830 2012


1826 ENDO12L_MON-255 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Deletion of Dicer and Pten in Mice Causes High-Grade Serous Cancers of the Female Reproductive Tract That Originate from the Fallopian Tube Jaeyeon Kim, Donna M Coffey, Chad J Creighton, Zhifeng Yu, Shannon M Hawkins, Martin M Matzuk Baylor College of Medicine, Houston, TX; The Methodist Hospital and Weill Medical College of Cornell University, Houston, TX High-grade serous carcinoma is the most common and deadliest ovarian cancer, causing 70% of all ovarian cancer deaths. To our dismay, it remains unknown how this cancer initiates and metastasizes before killing the woman. In the last decade, however, a hypothesis has emerged that the fallopian tube [ndash] not the ovary [ndash] is the primary origin of high-grade serous ovarian cancer. In our study, we provide direct evidence to the fallopian tube origin of this deadly serous cancer. Using mice carrying Amhr2-driven Cre recombinase, we have conditionally disabled in the reproductive tract two critical genes: Dicer, an essential gene for microRNA synthesis, and Pten, a tumor suppressor that negatively regulates the PI3K pathway. These double knockout (DKO) mice develop high-grade serous carcinomas that arise from the fallopian tube. After spreading to engulf the ovary, these serous cancers aggressively metastasize throughout the abdominal cavity [ndash] such as the mesentery, the pancreas, and the peritoneal membrane, most notably overlying the diaphragm [ndash] inducing ascites and ultimately killing 100% of the mice by 13 months. Besides the phenotypic and histological resemblance to human cancer, global gene expression analyses using microarrays show that these mouse serous carcinomas highly express numerous known markers of high-grade serous ovarian cancer in humans, also demonstrating molecular similarities. In addition, surgically removing ovaries, unilaterally or bilaterally, does not prevent these DKO mice from developing serous carcinomas, but after removing the fallopian tubes and leaving the ovaries intact, the DKO mice fail to produce the tumors [ndash] confirming the fallopian tube origin of the serous carcinomas. Finally, analysis of the DKO mice at early time points reveals that these serous carcinomas begin in the stroma of the fallopian tube, suggesting that the cancers arise through a mesenchymal-to-epithelial transition (MET), a novel mechanism of carcinoma initiation. This study thus presents a new paradigm for the origin and initiation of high-grade serous ovarian cancer.[br][br]Sources of Research Support: National Cancer Institute; Ovarian Cancer Research Fund; Baylor College of Medicine Partnership; and National Institute of Health Ruth L. Kirschstein National Research Service Award.[br][br]Nothing to Disclose: JK, DMC, CJC, ZY, SMH, MMM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1316 302 2162 MON-255 PO43-01 Monday 1831 2012


1827 ENDO12L_MON-256 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Defining the Roles of Cellular Senescence in Ovarian Tumorigenesis Michael D Fountain, Stephanie A Pangas, Keiichi I Nakayama, Philipp Kaldis, Martin M Matzuk Baylor College of Medicine, Houston, TX; Kyushu University, Fukuoka, Japan; Institute of Molecular and Cellular Biology, Proteos, Singapore; Baylor College of Medicine, Houston, TX Ovarian cancer is the most lethal gynecologic malignancy and the fifth most common cause of cancer death in women. Based on previous studies, defects in cellular senescence have been implicated as critical to understanding the pathogenesis of cancer. The ability of senescent cells to cease division is an important response to block the proliferation of cancer cells. Moreover, dysfunction in senescence pathways can lead to a decreased ability for cells to be removed and/or allow for aberrant proliferation. Our lab has been studying ovarian cancer in a mouse model lacking inhibin [alpha] ([italic]Inha)[/italic]. These [italic]Inha[/italic] null mice exhibit dysregulation in important cell-cycle complexes and cellular senescence regulators, and develop fatal ovarian granulosa cell tumors. S-phase kinase protein 2 (SKP2), an E3-ubiquitin ligase, marks p27 and p21 for degradation. Degradation of these tumor suppressors allow for the up-regulation of CDK2 and the subsequent G1 to S-phase transition, generating proliferation and cell growth. Our microarray data showed that multiple genes encoding cell senescence regulators are upregulated in [italic]Inha[/italic] null granulosa cells prior to cancer formation. This information lends the hypothesis that cellular senescence pathways regulated specifically by SKP2, and proteins upstream (MYC family members) and downstream (cyclin/CDK complexes), control the development of granulosa cell cancers. [bold]The central hypotheses are that: 1) Cellular senescence in the ovary controls the balance of apoptosis, proliferation, and differentiation of ovarian cancer; and 2) Alterations in key components of senescence (i.e., SKP2, cyclinA2/E1/E2/CDK2) can influence ovarian tumorigenesis. [/bold]To test these hypotheses, we plan to: (1) Elucidate the effects of cellular senescence on tumorigenesis and pathophysiology of gonadal tumors, and (2) Delineate the molecular mechanisms of cellular senescence in gonadal tumorigenesis. Double knockout (DKO) female mice are being created to study cellular senescence and cell-cycle regulation in granulosa cell proliferation and differentiation of gonadal tumorigenesis [italic]in vivo[/italic].[br][br]Sources of Research Support: This research is supported by NIH grant HD32067 from the Eunice Kennedy Shriver National Institute of Child Health and Human Development.[br][br]Nothing to Disclose: MDF, SAP, KIN, PK, MMM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2264 302 2163 MON-256 PO43-01 Monday 1832 2012


1828 ENDO12L_MON-257 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Nuclear Receptor CAR Overcomes Wnt/[beta]-Catenin Induced Liver Growth Arrest and Promotes Liver Tumorigenesis Bingning Dong, Ju-Seog Lee, David D Moore Baylor College of Medicine, Houston, TX; The University of Texas MD Anderson Cancer Center, Houston, TX Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide. The limited effectiveness of standard chemotherapeutic approaches makes HCC the third leading cause of cancer related mortality (1-4). Aberrant activation of Wnt/[beta]-catenin contributes to 34% of human HCC (5). However, [beta]-catenin activation alone is not sufficient to induce liver cancer in mouse models, indicating that other genetic events are need for tumorigenesis (6). Interestingly, about 80% of the tumors produced in the widely used diethylnitrosamine (DEN; carcinogen and initiator) plus phenobarbital (PB; promoter) mouse liver tumor model carry activating mutations in [beta]-catenin, while tumors from mice treated with DEN alone do not (7). The nuclear receptor CAR is one of the primary regulators of drug metabolism and is essential for PB tumor promotion (8). Thus, we hypothesize that CAR and [beta]-catenin coordinately promote liver tumorigenesis.[br]To explore the functional interaction between [beta]-catenin and CAR, we combined liver specific activation of both CAR (induced by TC, a CAR ligand) and [beta]-catenin (AdCre-mediated deletion of inhibitory exon 3 in ctnnb1[sup]loxP/loxP(ex3)[/sup] mice). We measured liver size, hepatocyte proliferation, and gene expression at different time points. Our data shows that activation of [beta]-catenin alone initially promotes liver growth but eventually leads to p53 response and cellular senescence. Activation of CAR alone also induces rapid, but limited liver growth. Remarkably, combined activation of CAR and [beta]-catenin induces uncontrolled liver growth (up to 25% liver/body weight ratio) which results in lethality, despite the maintenance of normal liver function. CAR activation overcomes the blockade to [beta]-catenin induced via induction of MDM2 and inactivation of the p53 dependent oncogene induced senescence pathway. In mice infected with lower titer of AdCre, the lethal phenotype of whole liver growth is absent, long-term activation of CAR and [beta]-catenin is sufficient to drive liver tumorigenesis. Thus, we conclude that CAR activation overcomes [beta]-catenin induced liver growth arrest and coordinately promotes liver tumorigenesis. Microarray and bioinformatic comparison studies demonstrate that our novel liver tumor model closely recapitulates human HCC with [beta]-catenin activation. Thus, our model provides a powerful tool to study the mechanisms, prevention, and treatment of [beta]-catenin associated liver cancer.[br][br](1). El-Serag, H.B., Marrero, J.A., Rudolph, L., and Reddy, K.R. 2008. Diagnosis and treatment of hepatocellular carcinoma. Gastroenterology 134:1752-1763. (2). Llovet, J.M., Ricci, S., Mazzaferro, V., Hilgard, P., Gane, E., Blanc, J.F., de Oliveira, A.C., Santoro, A., Raoul, J.L., Forner, A., et al. 2008. Sorafenib in advanced hepatocellular carcinoma. N Engl J Med 359:378-390. (3). Pellicano, R., El-Serag, H.B., and Rizzetto, M. Hepatocellular carcinoma at the cutting edge of science and practice. Minerva Med 101:391-393. (4). Severi, T., van Malenstein, H., Verslype, C., and van Pelt, J.F. Tumor initiation and progression in hepatocellular carcinoma: risk factors, classification, and therapeutic targets. Acta Pharmacol Sin 31:1409-1420. (5). de La Coste, A., Romagnolo, B., Billuart, P., Renard, C.A., Buendia, M.A., Soubrane, O., Fabre, M., Chelly, J., Beldjord, C., Kahn, A., et al. 1998. Somatic mutations of the beta-catenin gene are frequent in mouse and human hepatocellular carcinomas. Proc Natl Acad Sci U S A 95:8847-8851. (6). Harada, N., Miyoshi, H., Murai, N., Oshima, H., Tamai, Y., Oshima, M., and Taketo, M.M. 2002. Lack of tumorigenesis in the mouse liver after adenovirus-mediated expression of a dominant stable mutant of beta-catenin. Cancer Res 62:1971-1977. (7). Aydinlik, H., Nguyen, T.D., Moennikes, O., Buchmann, A., and Schwarz, M. 2001. Selective pressure during tumor promotion by phenobarbital leads to clonal outgrowth of beta-catenin-mutated mouse liver tumors. Oncogene 20:7812-7816. (8). Qatanani, M., and Moore, D.D. 2005. CAR, the continuously advancing receptor, in drug metabolism and disease. Curr Drug Metab 6:329-339.[br][br]Sources of Research Support: This work is supported by NIH R01 grant (DK-46546).[br][br]Nothing to Disclose: BD, J-SL, DDM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2118 302 2164 MON-257 PO43-01 Monday 1833 2012


1829 ENDO12L_MON-258 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Regulation of Prostate Cancer by a Steroid-Sulfating Sulfotransferase Nooshin Mirkheshti, Soyoung Kim, Rosario Mendez-Meza, Komal Arora, Marlo Nicolas, Sherry Dodd, Barbara Christy, Chung Song, Bandana Chatterjee University of Texas Health Science Center at San Antonio, San Antonio, TX; University of Texas Health Science Center at San Antonio, San Antonio, TX; South Texas Veterans Health Care System, San Antonio, TX Sulfation of DHEA (dehydroepiandrosterone) restricts its oxidative conversion to androstenedione, thus limiting testosterone and dihydrotestosterone production in the androgen biosynthesis pathway. DHEA sulfation in the prostate is mediated by the sulfotransferase SULT2B1b and thus, the prostate androgen pool may be partly regulated by the level and activity of SULT. In keeping with this, we observed significant acceleration in the proliferation rate of SULT2B1b-knocked down prostate cancer cells upon supplementation of the steroid-stripped media with DHEA. Hence, we postulated that low SULT expression, and an attendant anticipated rise in the tissue androgen level, would promote prostate cancer in a clinical setting. Here we report that the SULT2B1b level is markedly reduced in the prostate cancer tissue compared to its level in the matched non-cancer adjacent tissue. The cancer associated SULT reduction was evident from immunohistochemical as well as immunoblot analyses of multiple prostatectomy specimens using an in-house produced antibody that is specific to SULT2B1b. We further show that this SULT isoform is induced in multiple human prostate cancer cell lines and in the mouse prostate by calcitriol (1,25-dihydroxy vitamin D[sub]3[/sub]) as a result of transcriptional stimulation by the D[sub]3[/sub] receptor (VDR) and its RXRa partner. The VDR/RXRa heterodimer binds to a directly repeated DNA sequence in the upstream [italic]SULT [/italic]promoter, as revealed by chromatin immunoprecipitation, gel mobility shift and DNAse1 footprinting assays. Point mutations in the D[sub]3[/sub]-responsive cis element abrogated SULT induction. These results suggest that the elevated prostate SULT2B1b level may partly account for the calcitriol-mediated inhibition of prostate cancer [ndash] an effect that has been widely reported in cell culture and xenograft tumor models. Reduction of the prostate androgen pool by inhibiting CYP17A1, the rate limiting enzyme in androgen biosynthesis, has proved to be a viable strategy for inhibiting advanced prostate cancer that fails conventional chemotherapy. However, development of resistance against CYP17A1 inhibition is commonly observed. We suggest that the DHEA sulfation pathway possibly offers an alternative target that can be exploited to impede disease progression. Additionally, the SULT2B1b level in biopsied specimens may serve as a marker for prostate cancer prognosis. Studies are under way in our laboratory to explore these possibilities.[br][br]Sources of Research Support: A VA Merit Review grant, by NIH and by a supplemental funding from Translational Technology Resource at UTHSCSA.[br][br]Nothing to Disclose: NM, SK, RM-M, KA, MN, SD, BC, CS, BC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1307 302 2165 MON-258 PO43-01 Monday 1834 2012


1830 ENDO12L_MON-259 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Dickkopf 4 Positively Regulated by the Thyroid Hormone Receptor and Inhibited Cell Migration Chen-Hsin Liao, Kwang-Huei Lin Chang Gung University, Taoyuan, Taiwan Thyroid hormone (T3) mediates cellular growth, development, and differentiation by binding to the nuclear thyroid hormone receptor (TR). Recent studies suggest that long-term hypothyroidism is associated with human hepatocellular carcinoma (HCC) independent from other major HCC risk factors. DKK4, a secreted protein, antagonizes the Wnt signal pathway. Previously, we demonstrated that T3 plays a suppressor role by inducing DKK4 expression in HCC cells at both the mRNA and protein levels. Moreover, Over-expression of DKK4 in HCC cells leading to antagonize canonical Wnt signaling pathway. However, the underlying T3-regulated DKK4 mechanism is still unknown. The 5[apos]promoter region was serial deleted and reporter assay was performed to localize the T3-response element (TRE). An atypical direct repeated TRE between nucleotides [ndash]1645 and [ndash]1629 confers T3 responsiveness to the DKK4 gene. This region was further confirmed by chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assay (EMSA) assays. Further, stable DKK4 over-expression in SK-Hep-1 cells decreased cell invasion and metastasis potential in vivo as well as in vitro by reducing the matrix metalloproteinase-2 (MMP-2) expression. Taken together, this study suggests that DKK4, which is directly regulated by T3, has an inhibitory function in cell invasion, migration, and tumor progression.[br][br]Sources of Research Support: This work was supported by grants from Chang-Gung University, Taoyuan, Taiwan (CMRPD 34013, NMRP 140511) and from the National Science Council of the Republic of China (NSC 94-2320-B-182-052).[br][br]Nothing to Disclose: C-HL, K-HL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 120 302 2166 MON-259 PO43-01 Monday 1835 2012


1831 ENDO12L_MON-260 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) EW-7197, a Novel ALK5 Inhibitor Inhibited Lung Metastasis of Breast Cancer Cells Both [italic]In Vitro[/italic] and Mouse Model [italic]In Vivo[/italic] Seung W Kim, Sol J Kim, Sang A Park, Min J Kim, Jee Y Son, Dae-Kee Kim, Yhun Y Sheen Ewha Womans University, Seoul, Korea We have synthesized new ALK5 inhibitor, EW-7197 and investigated their effects of on breast cancer cells as well as tumor bearing MMTV/cNeu transgenic mouse, 4T1 or 4T1-Luc cell-orthotopic implanted mouse models. The level of [beta]-casein or the number of metastatic nodule in the lung of the breast tumor bearing MMTV/cNeu transgenic mice or 4T1 cell-orthotopic implanted mice, respectively, were decreased by EW-7197 treatment. The treatment of the EW-7197 also inhibited the luciferase activity in the lung of 4T1-Luc cell-orthotopic implanted mouse compared to that of untreated animal. The levels of MMP2 and MMP9 were decreased in the lung and breast tumor of the EW-7197 treated mice. To determine the inhibitory effect of EW-7197 on TGF-[beta]1-induced transcriptional activation, we performed luciferase reporter gene assay in breast cancer cells. EW-7197 effectively blocked TGF-[beta]1-induced luciferase activity in the cells in a dose-dependent manner. TGF[beta] signaling, the phosphorylation of smad2/3 was decreased by EW-7197. TGF[beta]-stimulated mesenchymal markers of epithelial mesenchymal transition (EMT) were inhibited by ALK5 inhibitor concomitant treatment. Furthermore, EW-7197 rescued TGF-[beta][ndash]induced loss of E-cadherin in NMuMG cells. And also, EW-7197 showed higher inhibition on cell motility and cell invasion based on wound healing assay or invasion assay than SB-505124 and LY-2157299. In summary, EW-7197 has anti-metastatic effect on breast cancer cells [italic]in vitro[/italic] as well as breast cancer models [italic]in vivo[/italic]. This data strongly argue that the ALK5 inhibitor have anti-metastasis effects via inhibiting ALK5 activity [italic]in vivo[/italic] and [italic]in vitro[/italic].[br][br]Nothing to Disclose: SWK, SJK, SAP, MJK, JYS, D-KK, YYS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1267 302 2167 MON-260 PO43-01 Monday 1836 2012


1832 ENDO12L_MON-261 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Identification of Preoperative Diagnostic Biomarkers for Thyroid Cancer Arturs Abols, Kristine Ducena, Diana Andrejeva, Janis Vilmanis, Zenons Narbuts, Juris Tars, Aija Line, Valdis Pirags Latvian Biomedical Research and Study Centre, Riga, Latvia; University of Latvia, Riga, Latvia; Pauls Stradins Clinical University Hospital, Riga, Latvia; Riga Eastern Clinical University Hospital, Riga, Latvia; University of Latvia, Riga, Latvia Currently, the cytological examination of the fine needle aspiration biopsy (FNAB) is the standard pre-operative diagnostic method for discriminating between benign and malignant thyroid nodules, however in [sim]20% of cases the results are non-informative due to the lack of conclusive morphological evidence of malignancy.The aim of the current study is to identify miRNAs that could serve as molecular biomarkers of thyroid cancer and could improve the diagnostic performance of FNAB. In order to identify candidate miRNAs that are overexpressed in thyroid cancer we determined miRNA expression profiles in the training set of 4 specimens of papillary thyroid carcinomas (PTC), 3 follicular thyroid carcinomas (FTC), 2 medullary carcinomas (MTC) and 2 anaplastic thyroid cancers (ATC), 4 benign adenomas and pooled relatively normal thyroid tissues as a reference using Exiqon[apos]s microRNA Array, v.11.0. This resulted in the identification of 11 miRNAs (miR- 1, miR-133a, miR-133b, miR-146b-5p, miR-146a, miR-182, miR-181a, miR-153, miR-155 and miR- PlusE1031) that were overexpressed in one or more of thyroid cancer subtypes comparing with normal thyroid tissues and benign nodules. The candidate miRNAs were tested in an independent cohort of samples (M=35; B=35; N=30) byqRT-PCR that showed that 3 of them (miR-146b-5p; miR-181a; miR-155) where statistically significantly overexpressed in cancers compared to benign nodules and normal tissues (p[lt]0,05). ROC curve analysis showed that AUC for these miRNAs as individual markers for discriminating between malignant vs benign nodules ranged from 0,61 to 0,70. Next, we applied a multivariate logistic regression analysis to develop a biomarker model combining these miRNAs and 6-gene mRNA expression signature that we have identified previously. This resulted in the multiplex model that was based on the expression analysis of LGALS3, TFF3, BIRC5, CDH1, DPP4, miR-146b-5p, miR-155 and miR-153 and had AUC of 0,96; p[lt]0,0001; CI 95%=0,91-0,99. In conclusion, this study resulted in the discovery of 8 biomarker model that outperforms the currently known biomarkers of thyroid malignancies and clearly warrants feasibility studies on FNAB specimens and the validation study in the multi-centre setting.[br][br]Sources of Research Support: Latvian Science Foundation grant 2009-2013.[br][br]Nothing to Disclose: AA, KD, DA, JV, ZN, JT, AL, VP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 814 302 2168 MON-261 PO43-01 Monday 1837 2012


1833 ENDO12L_MON-262 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Development and Validation of a LC-MSMS Method for the Simultaneous Determination of Urinary Steroids for Etiologic Diagnosis of Essential Hypertension and/or Metabolic Syndrome Fidel Allende, Sandra Solari, Carmen E Campino, Cristian A Carvajal, Rene Baudrand, Carlos F Lagos, Carolina Valdivia, Gareth Owen, Andrea Vecchiola, Carlos E Fardella School of Medicine, Faculty of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile; School of Medicine, Faculty of Medicine, Pontificia Universidad Catolica de Chile, Santiago, Chile; Faculty of Biological Sciences, Pontificia Universidad Catolica de Chile, Santiago, Chile; Pontificia Universidad Catolica de Chile, Santiago, Chile Cortisol (F) homeostasis is important in maintaining blood pressure and its dysregulation could be implied in hypertension (HT) and metabolic syndrome (MetS). F availability is modulated by two enzymes; 11[beta]-HSD1 which preferentially converts the inactive cortisone (E) to active F, while 11[beta]-HSD2 transforms F to E, protecting the mineralocorticoid receptor from erroneous activation by F. In contrast, the 5[alpha] and 5[beta] reductases (with 3 [alpha]-HSD) inactivate cortisol to its tetrahydrometabolites: Allo-THF, THF and THE. A subtle and local F increase could be detected by measuring 24 h urine metabolites being liquid chromatography tandem mass spectrometry (LC/MSMS) the preferred reference method. [bold]Aim:[/bold] Develop and validate using FDA parameters a LC/MSMS method for the simultaneous determination of F, E, Allo-THF, THF and THE in a clinical laboratory for better understanding HT and/or MetS pathogenesis. [bold]Methods:[/bold] Steroid-free urine enriched with known concentrations of each analyte were used for method optimization and validation. Steriods were extracted from 1 mL of urine, using D4-Cortisol as internal standard, before analysis by LC/MSMS (ABSCIEX, API4000). For clinical validation, urine from 83 HT patients and 77 controls (NT) were analyzed. The 11[beta]-HSD2 activity was evaluated by the F/E ratio and the 11[beta]-HSD1 by (AlloTHF+THF)/THE ratio. Activity was considered altered when the ratio exceeded mean+2SD of NT. [bold]Results:[/bold] The quantification range was 1-200 ng/mL with [gt]89% recovery for all analytes. Limit of detection, lower limit of quantification, coefficient of variation and accuracy were 0.2 ng/mL, 1.0 ng/mL, [lt]10% and 87-104% respectively. Methodology allowed the quantification of all 5 steroids in clinical samples. High F/E ratio was found in 6/83 HT (7%) suggesting a deficit in 11[beta]-HSD2 activity. Of the 77 HT with normal F/E ratio, 13 (17%) showed elevated (Allo-THF +THF)/THE ratio suggesting a 11[beta]-HSD1 overactivity. [bold]Conclusion:[/bold] Our LC/MSMS method is accurate, precise and reproducible and in accordance with FDA guidelines. Good sensitivity, recovery and matrix effect was obtained for the simultaneous measurement of the five analytes. This method allows the assessment of 11[beta]-HSD2 and 11[beta]-HSD1 activity in a single analytical run, thus providing an innovative tool in clinical endocrinology to explain the etiology of misclassified essential hypertension and/or metabolic syndrome.[br][br]Sources of Research Support: FONDEF D08I1087, FONDECYT 1100356 IMII P09/016-F grants. CFL and CAC are CONICYT PhD fellows.[br][br]Nothing to Disclose: FA, SS, CEC, CAC, RB, CFL, CV, GO, AV, CEF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 238 302 2169 MON-262 PO43-01 Monday 1838 2012


1834 ENDO12L_MON-263 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Associations between Body Mass Index and Thyroid Cancer Pathology Hye Jeong Kim, Se Won Kim, Yoon Young Cho, Sun-Mi Park, Ji Young Joung, Ji Cheol Bae, Jae Hyeon Kim, Sun Wook Kim, Jae Hoon Chung Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea [bold]Background: [/bold]Epidemiological studies have demonstrated that excess weight increases the risk of thyroid cancer. However, excess weight is associated with prognostic factors for thyroid cancer is uncertain. We evaluated the relationships between body mass index (BMI) and clinicopathological features of papillary thyroid cancer (PTC).[br][bold]Methods: [/bold]We reviewed the medical records of 2,064 patients who underwent total thyroidectomy for PTC. Demographic data, clinicopathologic features and metabolic parameters were recorded. BMI grouping (underweight or normal weight, overweight and obesity) were based on standardized categories set by WHO. Logistic regression models were used to assess the relation between BMI and clinicopathologic features of PTC.[br][bold]Results: [/bold]Positive associations were identified between BMI and tumors larger than 1cm (p[lt]0.0001), and between BMI and extrathyroidal invasion (p=0.017). Adjusted odd ratios (AORs) in Overweight (25.0-29.9 kg/m[sup]2[/sup]) and obesity ([ge]30 kg/m[sup]2[/sup]) for tumors larger than 1cm were 1.46 (95% CI, 1.18-1.81) and 2.05 (95% CI, 1.25-3.34), respectively. AORs in Overweight and obesity for extrathyroidal invasion were 1.27 (95% CI, 1.02-1.58) and 1.66 (95% CI, 1.01-2.72), respectively. No significant results were obtained between BMI and multifocality, lymph node metastasis, or distant metastasis.[br][bold]Conclusion: [/bold]Higher BMI is associated with tumor size and extrathyroidal invasion of PTC. This study lends support to the hypothesis that excess weight is associated with aggressive thyroid cancer.[br][br]Nothing to Disclose: HJK, SWK, YYC, S-MP, JYJ, JCB, JHK, SWK, JHC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1039 302 2170 MON-263 PO43-01 Monday 1839 2012


1835 ENDO12L_MON-264 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) NRAS Gene Mutation Is Associated with Papillary Thyroid Carcinoma Earlier Development Aline Carolina De Nadai Silva, Natassia Elena Bufalo, Lucas Leite Cunha, Marcio Jose Silva, Laura Sterian Ward University of Campinas, Campinas, Brazil; University of Campinas, Campinas, Brazil Environmental, dietary factors, and genetic changes have been implicated in the increased incidence of thyroid cancer observed all over the world in the last decades. The oncogenetic bases of this cancer have been extensively studied, and a number of mutually exclusive genetic alterations that activate the MAPK pathway are demonstrated in about 2/3 of the papillary thyroid carcinomas (PTC). The mutated genes that affect these pathways encode intracellular signal transducers BRAF and RAS. Mutations of the RAS genes are found in both benign and malignant thyroid tumors, suggesting that these mutations may occur early in the process of thyroid cell transformation, whereas BRAF mutation is associated with the morphological and functional alterations characteristic of the PTC phenotype. In order to better understand the relationship between these genes and PTC arising, we used SNP TaqMan Genotyping technique to identify BRAF V600E, NRAS and HRAS codon 61 mutations in 248 thyroid nodules patients (207 females and 41 males, 46.5 [plusmn] 14.24 years old) including 88 malignant nodules (86 PTC and 02 follicular carcinomas) and 159 benign nodules (99 hyperplastic nodules and 60 follicular adenomas). All patients were submitted to a standard routine diagnostic protocol and follow up management. BRAF and NRAS mutants were observed in 50 and 03 PTC, 01 and 0 follicular carcinomas, 0 and 09 benign nodules, respectively. None of the cases was mutated for HRAS gene. The inheritance of V600E BRAF gene was exclusive of PTC cases who were distinguished from benign nodules (p [lt]0.001) with a sensitivity = 58.62%, specificity = 100%, positive predictive value (PPV) = 100%, negative predictive value (NPV)= 81.83% and was not associated to PTC features of aggressiveness or outcome. NRAS neither appeared useful as a diagnostic marker, nor a prognostic marker. However, we observed that individuals who inherited the homozygous wild type (TT) NRAS codon 61 gene developed differentiated thyroid cancer (DTC) later (47.15 [plusmn] 14.18 years) than individuals who inherited a CT NRAS genotype (34.08 years [plusmn] 10:32), suggesting the occurrence of a disease anticipation phenomenon of around 12 years (p [lt]0.00186). In conclusion, we confirmed the literature concerning the importance of BRAF and RAS mutations and demonstrated that the inheritance of a heterozygous genotype in NRAS gene is related with the age of PTC diagnosis.[br][br]Sources of Research Support: FAPESP, CAPES.[br][br]Nothing to Disclose: ACDNS, NEB, LLC, MJS, LSW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1844 302 2171 MON-264 PO43-01 Monday 1840 2012


1836 ENDO12L_MON-265 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Growth Hormone Granulation Pattern and Somatostatin Receptor Subtype 2A Correlate with Postoperative Somatostatin Receptor Ligand Responses Jessica Brzana, Chris Yedinak, Sakir Gultekin, Johnny Delashaw, Maria Fleseriu Oregon Health [amp] Science University, Portland, OR; Oregon Health [amp] Science University, Portland, OR; Oregon Health [amp] Science University, Portland, OR; Oregon Health [amp] Science University, Portland, OR Introduction: Acromegaly is associated with serious morbidity and mortality if not well controlled. Somatostatin receptor ligands (SRLs), a mainstay of medical therapy, have predominant affinity for SSRT2. Therefore, we assessed whether tumor aggressiveness correlates with SRL sensitivity using defined tumor markers.[br]Methods: Retrospective review of 86 consecutive acromegaly surgeries (2006-2011: 70 patients, 11/86 cases were excluded for inadequate data). Immunohistochemical staining patterns: sparsely granulated, densely granulated, mixed GH-PRL and SSRT2 positivity (+) were correlated with clinicopathologic features, tumor recurrence, and somatostatin receptor ligand treatment response. Two-tailed t test, univariate ANOVA and bivariate correlation was performed using PAWS 18.[br]Results: Immunohistochemical staining: 22 densely granulated, 14 sparsely granulated, 23 mixed GH-PRL. SSTR2+: 20/20 (100%), 6/12 (50%), 19/20 (95%), respectively in each group, p[lt]0.001. Tumor size at diagnosis: densely granulated, sparsely granulated, mixed GH-PRL (19.2[plusmn]10.6mm, 21.6[plusmn]8.1mm, vs. 14.9[plusmn]7.8mm, p=0.033). Sparsely granulated tumors were diagnosed at younger age than densely granulated and mixed adenomas (41[plusmn]13y vs 52[plusmn]16y, and 49[plusmn]16y, p=0.146). Female prevalence: sparsely granulated [gt] mixed GH-PRL [gt] densely granulated (79%, 78% vs. 54%, (r=-.300, p=.034). Surgical cure was similar for all groups (sparsely granulated 50%, densely granulated 55%, mixed GH-PRL 52%). Tumor recurrence was higher for sparsely granulated vs. densely granulated (28.6% vs 13.6%, p=0.27). Response rates to SRL: 87% densely granulated, 66% sparsely granulated, 63% mixed GH/PRL (r=-663, p[lt]0.001). Overall response to SRLs was 13/16 (81%) in SSTR2+, and 0/2 in SSTR2[ndash] tumors (p= 0.016).[br]Discussion: Sparsely granulated adenomas are significantly larger at diagnosis, exhibit lower SSTR2 positivity and lower response to SRLs. Overall, patients with SSTR2+ tumors responded better than SSTR2[ndash] tumors to SRLs treatment. Taken as a whole, the response rate of SSTR2+ tumors to somatostatin receptor ligands in our study (81%) was much higher than in the unselected acromegaly population (20-30%) (p[lt]0.0001).[br]Conclusions: Detailed knowledge of immunohistochemical SSTR2+ status and tumor GH granularity could predict somatostatin receptor ligand response. These immunoreactive markers should be assessed routinely on surgical specimens to assess SRLs responsiveness and tumor aggressiveness.[br][br]Nothing to Disclose: JB, CY, SG, JD, MF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 781 302 2172 MON-265 PO43-01 Monday 1841 2012


1837 ENDO12L_MON-266 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Imatinib Associated with Dacarbazine and Capecitabine for Metastatic Adrenocortical Carcinoma Treatment Joao E Bezerra, Paulo M Hoff, Ana O Hoff, Madson Q Almeida, Antonio M Lerario, Rachel S Riechelmann, Tulio E Pfiffer, Ana C Latronico, Berenice Bilharinho Mendonca, Maria Candida BV Fragoso ICESP, S[atilde]o Paulo, Brazil; Hospital das Cl[iacute]nicas da Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Background: Adrenocortical carcinoma (ACC) is a rare aggressive tumor associated with poor outcomes. Mitotane is usually the first therapeutic option for advanced disease. Cisplatin-based chemotherapy has been the treatment of choice for progressive disease. Encouraging results come from the combination of mitotane, cisplatin, etoposide and doxorubicin (EDP-M). However, this regimen offers significant toxicity. Imatinib, a selective inhibitor of bcr-abl, PRGFR [alpha] and c-kit, has been tested as single agent in a small cohort of ACC patients with no apparent benefit. However, promising outcomes were seen in a phase I trial of imatinib associated with dacarbazine (DTIC) and capecitabine (unpublished data). Methods: All consecutive ACC patients treated with this regime were included. Imatinib was given at 400mg/day continuously. DTIC 330mg/m2 on D1 to D5 and capecitabine 1000mg/m2 bid from D1 to D14 was given every 3 weeks. Because of limiting hematologic toxicity in the first two patients, capecitabine doses were reduced to 850mg/m2 bid. Six cycles of treatment were preplanned. Results: Since November 2010, 6 female patients, age ranging from 26 to 63 years, were treated. Four patients had had progressive disease with mitotane besides adequate therapeutic serum levels. Three of them had also been previously treated with EDP-M. Mitotane were maintained during imatinib in five patients. Three patients received only one cycle of therapy for the following reasons: grade 4 febrile neutropenia, grade 4 liver toxicity and progressive disease. No other grade 3/4 toxicity was reported. One patient completed the six preplanned cycles achieving complete response. Two patients are still been treated, but partial response and stable disease, respectively, after six weeks. Conclusion: In this small cohort, half of patients benefited from imatinib together with DTIC and capecitabine. Based on this result, we are conducting a phase II trial of this combination. The molecular target of imatinib in ACC is not clear and remains to be determined.[br][br]Nothing to Disclose: JEB, PMH, AOH, MQA, AML, RR, TEP, ACL, BBM, MCBVF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2114 302 2173 MON-266 PO43-01 Monday 1842 2012


1838 ENDO12L_MON-267 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Ribonucleotide Reductase Large Subunit (RRM1) Gene Expression May Predict Adjuvant Mitotane Efficacy in Adrenocortical Cancer: [italic]In Vivo[/italic] and [italic]In Vitro[/italic] Experiments Massimo Terzolo, Marco Volante, Martin Fassnacht, Ida Rapa, Antonina Germano, Silvia De Francia, Silviu Sbiera, Fulvia Claudia Daffara, Paola Sperone, Giorgio Vittorio Scagliotti, Bruno Allolio, Mauro Papotti, Alfredo Berruti University of Turin, Orbassano, Italy; University Hospital W[uuml]rzburg, W[uuml]rzburg, Germany PURPOSE.[br]Mitotane is the reference systemic therapy for adrenocortical carcinoma (ACC), but its mechanism of action and possible predictors of treatment response remain poorly defined. Our aim was to evaluate the gene expression of ribonucleotide reductase large subunit 1 (RRM1) and excision repair cross-complementation group 1 (ERCC1) in ACC as potential biomarkers for clinical outcome and response to mitotane.[br]EXPERIMENTAL DESIGN.[br]Forty-five and 47 tissue samples from two cohorts (Orbassano, Italy; Wurzburg, Germany) of completely resected ACC were centrally analyzed using Real Time PCR for RRM1 and ERCC1 expression. Fifty-four patients received surgery alone and 38 received adjuvant mitotane after surgery. Clinical and pathological features were highly comparable in the two series. H295R and SW-13 ACC cell lines were also used for pharmacological tests.[br]RESULTS.[br]ERCC1 gene expression was not associated to clinical outcome. In contrast, high RRM1 gene expression was associated to shorter disease-free and overall survival at both univariate and multivariate analysis. In patients with low RRM1 gene expression adjuvant mitotane significantly reduced the risk of recurrence [HR, 0.37 (0.15-0.87); p=0.016] thus prolonging disease-free survival, whereas this effect was lost in patients with high RRM1[HR, 1.07 (0.57-2.00); p=0.83]. In vitro mitotane induced strong up-regulation of RRM1 transcription (up to 25-fold increase) in mitotane-insensitive SW-13 cells but not in mitotane-sensitive H295R cells.[br]CONCLUSION.[br]Our in vivo and in vitro data indicate that RRM1 gene expression is functionally associated to mitotane sensitivity and may represents a marker predicting response to adjuvant mitotane in ACC patients.[br][br]Nothing to Disclose: MT, MV, MF, IR, AG, SDF, SS, FCD, PS, GVS, BA, MP, AB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 950 302 2174 MON-267 PO43-01 Monday 1843 2012


1839 ENDO12L_MON-268 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Influence of Smoking History on Gene Expression Profiles That Predict Breast Cancer Behavior James L Wittliff, Sarah A Andres, Ted S Kalbfleisch University of Louisville, Louisville, KY In contrast to previous studies that focused on cigarette smoking and risk of breast cancer occurrence, our emphasis is on the influence of tobacco smoking on breast cancer risk of recurrence and progression. Our goal is to combine knowledge of lifestyle behavior (smoking history) and molecular phenotype of the breast lesion to improve assessment of risk of recurrence. Genes, such as NAT2, NAT1, CYP1A1, COMT, BRCA1 and BRCA2, are reported to play a role the relationship between tobacco smoking and breast cancer risk (e.g., (1)). We also examined superoxide dismutase (SOD) which has been implicated in estrogen receptor negative breast cancer. Gene expression was investigated by microarray of RNA of carcinoma cells isolated by laser capture microdissection from 247 de-identified breast tissue biopsies and correlated with clinical outcome using our extensive IRB-approved database. Cross-platform validation confirmed correlations between gene expression analyzed by qPCR and microarray. Elevated NAT1 expression levels in breast cancers of confirmed smokers (n=66) were correlated with increased disease-free survival (DFS: P=0.08, 95% CI=0.91-4.79) and overall survival (OS: P[lt]0.04, 95% CI=1.06-7.27), which was not observed in breast cancers of non-smokers (n=99). Expression of either CYP1A1, COMT or SOD2 was equivalent in the breast cancers of both smokers and non-smokers. Interestingly, expression of BRCA1 was diminished in breast cancers from patients with a smoking history compared to those of nonsmokers. However, expression of either SOD1 or BRCA2 (at below median levels) was associated with improved survival outcomes of non-smoking breast cancer patients (SOD1: DFS P=0.01, 95% CI=0.18-0.67 and OS P=0.02, 95% CI=020-0.84; BRCA2: DFS P=0.01, 95% CI=0.22-0.80 and OS P=0.01, 95% CI=0.19-0.80), which was not observed in patients with a smoking history. Collectively, results from this investigation illustrate that although smoking may not be an independent factor determining breast cancer progression, exposure of a patient to tobacco smoke combined with certain molecular phenotypes of breast carcinoma may alter clinical behavior of this disease.[br][br](1) Morabia A, Environ Mol Mutagen 2002; 39:89.[br][br]Sources of Research Support: In part by a grant from the Phi Beta Psi Charity Trust.[br][br]Nothing to Disclose: JLW, SAA, TSK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2269 302 2175 MON-268 PO43-01 Monday 1844 2012


1840 ENDO12L_MON-269 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Adipokines Serum Profile Helps To Identify Patients under Higher Risk of Developing Differentiated Thyroid Cancer Marjory Alana Marcello, Antonio Calixto, Ana Carolina Junqueira Vasques, Bruno Geloneze, Laura Sterain Ward University of Campinas, Campinas, Brazil; University of Campinas, Campinas, Brazil Although there are solid evidences that obesity is a risk factor for the development of differentiated thyroid carcinoma (DTC), the mechanisms involved in obesity-related thyroid neoplasia remain poorly understood. Because recent studies have suggested that thyroid cancer is associated with insulin resistance and adipokines expression, we used ELISA assays to quantify serum adiponectin, leptin, resistin and ghrelin levels in DTC patients and healthy individuals, investigating their potential role as markers of susceptibility to DTC. We studied 92 (81 females and 11 males, 46.09+13.80 years old) DTC patients and 105 (92 females and 13 males, 47.13+6.65 years old) healthy controls, carefully matched for age, gender and body mass index (BMI) and classified into subgroups: eutrophic (BMI:18.5-24.9; 34 and 40 subjects, respectively in each group), overweight (BMI:25.0=29.9; 31 and 39 subjects) and obese (BMI[gt]30.0; 23 and 24 subjects). Both serum adiponectin and leptin levels were higher in controls (4.535+2.88[micro]g/ml and 14.180+14.660ng/ml, respectively) than in patients (2.505+0.9766 [micro]g/ml, p[lt]0.0001 and 9.940+3.202ng/ml, p=0.0013, respectively). On the contrary, resistin levels were higher in DTC patients (14.570+2.993ng/ml) than in controls (8.105+6.431ng/ml, p[lt]0.0001). Ghrelin levels did not differ significantly between groups (p=0.8657). A multiple logistic regression analysis confirmed low resitin levels as an independent risk factor for DTC (OR=1.13; p=0.0194) whereas adiponectin (OR=0.890929; p[lt]0.0001) and leptin (OR=0.984974; p[lt]0.0001) higher levels were independent protective factors. Adiponectin remained as a protective factor (OR=0.890787; p[lt]0.0001) and resistin as a risk factor (OR=1.04; p[lt]0.0001) among eutrophic individuals. In overweight individuals, adiponectin and leptin also acted as protective factors (OR=0.871282; p[lt]0.0001 and OR=0.985764; p[lt]0.0001, respectively). The same was observed among obese individuals: OR=0.884865; p[lt]0.0001 and OR=0.984101; p[lt]0.0001, respectively. In conclusion, our data suggest that the reduction in serum adiponectin and leptin protective levels may be linked to the development of DTC. In addition, higher levels of resistin, an adipokine related to insulin resistance, may be a crucial factor of risk for DTC in eutrophic individuals. Adipokynes serum profile may be useful in the identification of populations who may benefit more from preventive measures aiming to reduce weight and insulin resistance.[br][br]Sources of Research Support: CNPq.[br][br]Nothing to Disclose: MAM, AC, ACJV, BG, LSW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 966 302 2176 MON-269 PO43-01 Monday 1845 2012


1841 ENDO12L_MON-270 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Inflammatory Cytokines CXCL1 and -2 Are Regulated by Progesterone and Calcitriol in Cancer Cells Leyla Kavandi, Michael Aaron Collier, Huyen Nguyen, Viqar Syed Uniformed Services University of the Health Sciences, Bethesda, MD Chronic inflammation has been shown to play a critical role in growth of cancers. NF-[kappa]B is a transcriptional factor that regulates diverse biological processes, including cell proliferation, survival and inflammation. We have previously shown inhibition of endometrial and ovarian cancer cell growth with progesterone and calcitriol. In the present study, we evaluated the effect of progesterone and calcitriol on the expression of inflammatory genes. Using RT-PCR array of inflammatory cytokine and receptor genes, we found two inflammatory cytokines, CXCL1 and -2 (GRO-A and -B), downregulated by both treatments. PCR array results were validated by Western blotting. Knockdown of NF-kB resulted in a reduced expression of CXCL1 and -2 and the inhibitory effect of progesterone and calcitriol on the expression of cytokines was abrogated in NF-kB-silenced cancer cells. Silencing of IkB[alpha] increased the expression of CXCL1 and -2 in cancer cells, which can be attributed to the increased activation of NF-kB-p65, caused by the lack of its inhibitor. Progesterone and calcitriol-induced inhibition was abolished in IkB[alpha]-knockdown cells. Our results demonstrate that suppression of IkB[alpha] phosphorylation by progesterone and calcitriol contributes to the reduced expression of CXCL1 and -2. We investigated the effect of CXCL1 and -2 downregulation on metastasis-promoting genes and found a marked inhibition of CXCR4, COX2 and matrix metalloproteinases (MMP2 and MPP-9). Overall, our results indicate that progesterone and calcitriol downregulate NF-kB in cancer cells, leading to the suppression of metastasis, thus providing the molecular basis for the treatment of endometrial and ovarian cancer patients with progesterone and calcitriol.[br][br]Nothing to Disclose: LK, MAC, HN, VS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 46 302 2177 MON-270 PO43-01 Monday 1846 2012


1842 ENDO12L_MON-271 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Wild-Type TRP53 (p53) Promotes Ovarian Cancer Cell Survival Lisa K Mullany, Zhilin Liu, Kwong-Kwok Wong, Erin R King, JoAnne S Richards Baylor College of Medicine, Houston, TX; The University of Texas MD Anderson Cancer Center, Houston, TX Low-grade and high-grade epithelial ovarian cancers (EOCs) have a different pathogenesis, molecular phenotype and responses to conventional chemotherapy. The tumor repressor protein 53 ([italic]Trp53[/italic]; or [italic]p53[/italic]) is mutated in almost all high-grade serous adenocarcinomas (96%) whereas low-grade tumors express elevated levels of wild type [italic]Trp53.[/italic] Compared to human low-grade invasive serous ovarian carcinomas, EOCs in mutant mice similarly express elevated levels of wild type [italic]Trp53[/italic]. To investigate the functions of TRP53 in the mutant mouse ovarian surface epithelial (OSE) cells, [italic]Trp53[/italic] was conditionally deleted in [italic]Pten/Kras [/italic]mice. Gene profiling analyses using RNA prepared TRP53+ ([italic]Pten/Kras[/italic] ([italic]Trp53+[/italic])) and TRP53- ([italic]Pten/Kras(Trp53[/italic]-)) OSE cells revealed that hundreds of genes are regulated by TRP53. In the TRP53 +cells, wild type TRP53 controls or enhances the expression of genes regulating proliferation, DNA repair and mitotic activity and markedly decreases genes with tumor suppressor functions. Rather than activating cell cycle arrest or apoptosis, wild type TRP53 in the [italic]Pten/Kras(Trp53+)[/italic] OSE cells promotes the formation of papillary-like structures, cell migration, adhesion and invasion. By contrast, cells lacking [italic]Trp53[/italic] exhibit a less aggressive phenotype and gene expression profiles more like control OSE cells. Thus, we have unveiled [underline]a novel role for wild-type TRP53 as a major promoter of ovarian cancer cell survival, differentiation and migration. These results provide a new paradigm: wild type TRP53 at low levels of activity does not preferentially induce apoptotic or senescent related genes in the [italic]Pten/Kras[/italic]([italic]Trp53+[/italic]) cells.[/underline] To test this paradigm, purified TPR53+ positive and TRP53- negative mouse OSE cells and a low-grade human EOC cell line were exposed to nutlin-3a, a small molecule that disrupts the interactions of TRP53 with MDM2, and increases TRP53 activity dramatically. In TRP53+ mouse and human OSE cells, nutlin-3a stimulated TRP53 activity and markedly induced cell cycle arrest and apoptotic genes but reduced DNA repair and survival genes. Thus, TRP53 controls global, molecular changes that are dependent not only on the level of wild type TRP53 expression but also on its activation status. Nutlin-3a provides a promising therapeutic small molecular for managing low-grade ovarian cancer and other cancers where wild-type TRP53 is expressed.[br][br]Nothing to Disclose: LKM, ZL, K-KW, ERK, JSR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 279 302 2178 MON-271 PO43-01 Monday 1847 2012


1843 ENDO12L_MON-272 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Inhibition of the Phosphoinositide 3-Kinase/mTOR Pathway and Influence on the Viability of Human PTEN-Deficient Lipoma Cells Franziska Kassner, Gordian Schmid, Franziska Wilhelm, Antje Korner, Antje Garten, Wieland Kiess University Hospital Leipzig, Hospital for Children and Adolescents, Leipzig, Germany [bold]Background:[/bold] The PTEN Hamartoma Tumor Syndrome (PHTS) is a rare genetic disease with a wide range of symptoms due to mutations in the [italic]PTEN[/italic] gene, which codes for a phosphatase involved in the mTOR signaling pathway. The development of lipomatosis is one symptom of PHTS, which can lead to severe complications due to obstructions of the airway or digestive system. Rapamycin treatment, as so far only available therapeutical option besides surgery, has proved to be only partially successful.[bold][br]Aims:[/bold] We aimed to test [italic]in vitro[/italic] whether pharmacological inhibition of AKT, phosphoinositide(PI) 3-kinase or mTOR leads to reduced viability or the induction of apoptosis in human PTEN-deficient lipoma cells.[bold][br]Methods:[/bold] Cells from a lipoma of a patient with PHTS were established and maintained in long term culture. Cell differentiation was determined by Nile Red lipid staining and cell counting. Viability was assessed using the WST-1 assay and apoptosis was measured flow-cytometrically after annexin V/propidium iodide staining.[bold][br]Results: [/bold]Lipoma cells had a lifespan of 91 population doublings with a doubling time of 25 h. They preserved a capacity for adipocyte differentiation of 55.1[plusmn]4.2% for 29 population doublings as measured by the rate of lipid accumulating cells. PTEN mRNA and protein levels were decreased compared to controls and a constitutive phosphorylation of the kinase AKT was ascertained. Rapamycin as inhibitor of mTOR complex 1 decreased viability by 43.4[plusmn]1.9% and adipocyte differentiation by 72.7[plusmn]5.4% (100 nmol/L). However, no induction of apoptosis could be detected. Rapamycin analoga had a smaller effect on cell viability at the same concentration. mTOR complex 1/2 inhibitors pp242 and WYE-354 (both 1 [micro]mol/L) decreased cell viability by 47.1[plusmn]3.6% and 56.5[plusmn]3.0%, whereas the PI3-kinase inhibitor LY294002 (100 [micro]mol/L) had a greater effect with 77.9[plusmn]4.6% reduction. A slight induction of apoptosis by 7.9 [plusmn] 1.7 % was seen after incubation with LY294002 (100 [micro]mol/L), which increased to 61.8 [plusmn] 2.1% at 500 [micro]mol/L LY294002 compared to control. Perifosine, an AKT inhibitor currently involved in clinical phase III trials, reduced cell viability by 96.4[plusmn]0.6% and induced apoptosis by 84.5[plusmn]0.9% (100 [micro]mol/l).[bold][br]Conclusion:[/bold] Inhibition of AKT by perifosine decreased lipoma cell viability to the greatest extent and induced significant apoptosis. Perifosine may be of therapeutical value for patients with PHTS.[br][br]Sources of Research Support: German Research Council - KFO 152 [ldquo]Atherobesity[rdquo] to W.K., the LIFE program (Leipzig Interdisciplinary Research Cluster of Genetic Factors, Clinical Phenotypes and Environment) to W.K., the IFB (Integrated Research and Treatment Center AdiposityDiseases)to G.S., BMBF (Federal Ministry for Education and Research)to W.K.;the Kompetenznetz Adipositas-Verbund LARGE (TP01) and Kompetenznetz Diabetes to W.K.[br][br]Nothing to Disclose: FK, GS, FW, AK, AG, WK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 340 302 2179 MON-272 PO43-01 Monday 1848 2012


1844 ENDO12L_MON-273 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Multifunctional Transcription Factor TFII-I Is an Activator of BRCA1 Function Michihiro Tanikawa, Osamu Wada-Hiraike, Akira Shirane, Katsutoshi Oda, Kei Kawana, Shunsuke Nakagawa, Tetsu Yano, Shiro Kozuma, Yuji Taketani The University of Tokyo, Tokyo, Japan; Teikyo University, Tokyo, Japan Background:The TFII-I is a multifunctional transcriptional factor known to bind specifically to several DNA sequence elements and to mediate growth factor signalling. A microdeletion at the chromosomal location 7q11.23 encoding TFII-I and the related family of transcription factors may result in the onset of Williams-Beuren syndrome, an autosomal dominant genetic disorder characterised by a unique cognitive profile, diabetes, hypertension, anxiety, and craniofacial defects. Hereditary breast and ovarian cancer susceptibility gene product BRCA1 has been shown to serve as a positive regulator of SIRT1 expression by binding to the promoter region of SIRT1, but cross talk between BRCA1 and TFII-I has not been investigated to date. Methods:A physical interaction between TFII-I and BRCA1 was explored. To determine pathophysiological function of TFII-I, its role as a transcriptional cofactor for BRCA1 was investigated. Results:We found a physical interaction between the carboxyl terminus of TFII-I and the carboxyl terminus of BRCA1, also known as the BRCT domain. Endogenous TFII-I and BRCA1 form a complex in nuclei of intact cells and formation of irradiation-induced nuclear foci was observed. We also showed that the expression of TFII-I stimulates the transcriptional activation function of BRCT by a transient expression assay. The expression of TFII-I also enhanced the transcriptional activation of the SIRT1 promoter mediated by full-length BRCA1. Conclusion:These results revealed the intrinsic mechanism that TFII-I may modulate the cellular functions of BRCA1, and provide important implications to understand the development of breast cancer.[br][br]Nothing to Disclose: MT, OW-H, AS, KO, KK, SN, TY, SK, YT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1156 302 2180 MON-273 PO43-01 Monday 1849 2012


1845 ENDO12L_MON-274 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Expression of a Constitutively Active Prolactin Receptor Causes Histone Trimethylation of the p53 Gene in Breast Cancer Dunyong Tan, Ameae M Walker Jishou University, Jishou, China; University of California, Riverside, Riverside, CA Background: Prolactin (PRL) is a pituitary polypeptide hormone characterized by multiple biological actions including stimulation of growth in the prostate, and formation of secretory alveoli and stimulation of milk protein gene expression in the mammary gland. PRL exerts its effect by dimerizing its receptor (PRLR) on the plasma membrane, and regulating gene expression through the JAK-Stat signal pathway. We have previously described a natural variant of the PRLR in which the S2 subdomain of the extracellular domain is missing (Delta S2). Delta S2 PRLRs are dimerized in the absence of PRL and have constitutive activity in the promotion of breast cancer cell growth. Enhancer of Zeste homologue 2 (EZH2), as one of the histone modifying enzymes, is a key factor regulating gene expression by epigenetic modification. We hypothesized that these constitutive activated Delta S2 PRLRs played a pathogenic role in breast cancer in part through alterations in the expression of EZH2 and the trimethylation of histone 3 on lysine 27 (H3K27Me3). Materials and methods: In order to verify the clinical significance and to establish the link between Delta S2 PRLR expression and epigenetic change, EZH2, H3K27Me3 and Delta S2 PRLR were detected in both normal and cancerous human breast tissue. Also, overexpression of Delta S2 PRLR in breast epithelial cells was achieved by infection with adenovirus carrying the cDNA. Western blot, chromatin immunoprecipitation (ChIP assay) and acid histone extraction were applied to detect expression of EZH2 as well as trimethylation of histone 3, respectively. Results: In breast tissue, higher EZH2 expression and higher H3K27Me3 were found associated with higher Delta S2 expression in breast cancer samples. In breast epithelial cells, overexpression of Delta S2 PRLR increased EZH2 methyltransferase mRNA and protein, induced EZH2 methyltransferase recruitment to chromatin, increased the trimethylation of histone 3 on lysine 27 (H3K27Me3), and decreased expression of the p53 Gene. Conclusion: Delta S2 PRLR plays an important pathogenic role in breast cancer through epigenetic modification. Elevated expression of the Delta S2 PRLR, achieved by alternate splicing of the pre-mRNA of the full length form is a new mechanism contributing to human breast cancer.[br][br]Sources of Research Support: National Natural Science Foundation of China Grant 81172497; Jishou University Research Grant jsdxkyzz101010.[br][br]Nothing to Disclose: DT, AMW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1363 302 2181 MON-274 PO43-01 Monday 1850 2012


1846 ENDO12L_MON-275 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Deleterious Cross Talk between Estrogen and Thyroid Signaling in Breast Cancers Reema S Wahdan-Alaswad, Zeying Fan, Bolin Liu, Dhalia Ercan, Susan M Edgerton, Ann D Thor University of Colorado, Aurora, CO; Dana-Farber Cancer Institute, Boston, MA Long term, exogenous steroid hormone use promotes breast cancer, especially in post-menopausal women. The role of thyroid hormone (TH) and its receptor (TR) in breast cancer is less well defined. Up to 2/3 of women with breast cancer reportedly have clinical or subclinical thyroid disease. Estrogen receptor (ER) and TR are members of a nuclear receptor superfamily that binds to low molecular weight ligands, estrogen (E) and TH, respectively. They transduce signals and regulate gene transcription, through hormone response elements with significant structural homology. Patients on long term TH who develop breast cancer are most frequently postmenopausal, with ER+ tumors.[br]We studied over 750 archival, chemotherapy untreated node negative (LN-) breast cancer patients who were followed for a mean of 10.2yr, of which 56 received TH. TH administration was significantly associated with a worse disease free and overall survival (DFS and OS) in all patients, as well as ER+ but not the ER- patient subsets. For example, in ER+, LN- patients the 5 yr DFS was 91% in TH untreated patients, as compared to 69% in patients who received TH. Similarly, at 10 years the DFS was 84% in TH untreated, and 54% in TH treated patients. Our data suggests a deleterious interaction between tamoxifen and TH. The 5 year DFS was only 50%, in ER+ LN- patients receiving both tamoxifen and TH. These outcomes could not be explained on the basis of clinical or histologic features, and they remained significant using both univariate and multivariate statistical models. With hypothesized that cross talk between estrogenic and thyroid signaling, possibly involving ER at the tamoxifen binding site, may be responsible for these data.[br]We have compared several forms of TH, to determine if they had similar effects on ER+ breast cancer (BC) cells in vitro. Each significantly enhanced the growth of ER+, but not ER- BC cell lines. Combinatorial treatment with estrogen (E) and T3 or T4 were particularly potent in inducing ER positive breast cancer cell proliferation, in a dose and time dependent manner. TH resulted in more cell proliferation in ER+, p53 mutant cells (like T47D) as compared to ER+, p53 competent cells (like MCF7). TH upregulated Cyclins A, B, and E, E2F1 and activated MAPK/ERK1/2 but not AKT signaling. TH significantly abrogated tamoxifen associated growth inhibition, whereas this deleterious interaction was not seen with ICI 182,780 (fulvestrant) or the aromatase inhibitor Aromasin.[br][br]Sources of Research Support: Mary Kay Ash Foundation (AT, DE).[br][br]Nothing to Disclose: RSW-A, ZF, BL, DE, SME, ADT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1742 302 2182 MON-275 PO43-01 Monday 1851 2012


1847 ENDO12L_MON-276 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Metformin, Glucose Metabolism and Oncogenic Signaling in Breast Cancers Reema S Wahdan-Alaswad, Zeying Fan, Susan M Edgerton, Xin-Sheng Deng, Bolin Liu, Nicole Spoelstra, Jennifer K Richer, Ann D Thor University of Colorado Denver Anschutz Medical Campus, Aurora, CO Background: Chronic diseases associated with metabolic dysregulation include obesity and diabetes, which are independently associated with an increased risk and worse prognosis for breast cancer.[br]Hypothesis:We hypothesize that maintenance of glucose homeostasis may reduce breast carcinogenesis and promotion[br]Results: We have studied the effects of glucose, as a mitogenic and stress inducing agent, at various concentrations (at 5, 7. 10 or 17mM) on breast cancer cell lines. Hyperglycemia ([gt]5mM) promotes cell growth and colony formation, especially in luminal A and triple negative (as compared to HER2+) breast cancer cells. Lower glucose concentrations ([lt]5mM) activate cell stress pathways through key intermediate molecules, including cell surface receptors (EGFR, ER-alpha, IGF1R), signaling intermediates (Akt, mTor, AMPkinase, Stat3, MAPk, IRS2, Smads 2 and 3, and cell cycle regulators Cyclins A, B, D and E.[br]The majority of aforementioned biological and signaling effects were abrogated or significantly reduced by the addition of metformin to the media. Of interest, the EC50 of metformin was dependent on the glucose concentration. In general, the higher the glucose level, the higher the EC50 of metformin. To study early and late metformin associated gene expression changes, we used expression arrays on RNA derived from 5 breast cancer cell lines, grown at various glucose concentrations, for 6, 12 and 24 hours in vitro. Metformin resulted in more frequent shifts in gene expression when it was administered to cells grown at 5mM glucose, as compared to higher levels. In general, it downregulated more genes than it upregulated. Furthermore, metformin reduced mammosphere formation as well as the stem cell subpopulation (determined by the expression of CD44 (by one log scale) in flow cytometry. Metformin also inhibited cell motility in a panel of triple negative breast cancer cell lines, where it showed little inhibition of motility in luminal A type cells.[br]Conclusions: These data support a role for glucose in breast cancer mitogenesis, stem cell maintenance, and signaling. We believe that further study of metformin[apos]s effects on various molecular subtypes of breast cancers, both in vitro and in vivo, may provide significant and translationally relevant insights into breast cancer treatment paradigms and the use of metformin as an anti-breast cancer agent.[br][br]Sources of Research Support: The Komen Breast Cancer Foundation (ADT, JR ZF, SME, NS).[br][br]Nothing to Disclose: RSW-A, ZF, SME, X-SD, BL, NS, JKR, ADT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2138 302 2183 MON-276 PO43-01 Monday 1852 2012


1848 ENDO12L_MON-277 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Early Results of an [italic]In Vivo[/italic] Clinical Study Utilizing an Integrated FNA Biopsy Tool with Elastic Light Scattering Spectroscopy for the Evaluation of Thyroid Nodules Jennifer E Rosen, Hyunsuk Suh, Ilona D Goukassian, Eladio Rodriquez-Diaz, Ousama A[apos]amar, Irving J Bigio, Stephanie L Lee Boston Medical Center, Boston, MA; Boston University, Boston, MA; Boston Medical Center, Boston, MA [bold]Background: [/bold]Thyroid cancer is the most common endocrine malignancy with the fastest growing rate of all cancers. The current gold standard, fine-needle aspiration biopsy (FNAB), yields approximately 10-25% of indeterminate results, leading to thyroidectomy for diagnosis. Elastic scattering spectroscopy (ESS) is a new, minimally invasive optical-biopsy technique, mediated by a fiber-optic probe, which is sensitive to cellular and subcellular features (1). We assessed the potential to incorporate an ESS probe into preoperative transcutaneous aspiration biopsy to differentiate benign from malignant thyroid nodules.[br][bold]Methods:[/bold] We built a miniaturized ESS probe to fit through a 23-gauge biopsy needle and tested it with IRB approval on 42 patients undergoing ultrasound-guided FNAB of thyroid nodules. ESS data was collected from three distinct locations within the thyroid nodule. Cytology was our gold standard, except for indeterminate cytology when we used final post-surgery pathology.[br][bold]Results:[/bold] All patients tolerated the procedure well; the additional time required was less than 30 seconds. Spectral appearance differed between solid and liquid portions of the thyroid nodule. Only one patient had an insufficient biopsy (2.3%) and spectra could be collected on all patients. Examination of cytology obtained with the integrated ESS-FNA device compared to standard FNA biopsy showed similar numbers and types of cells. A waveform signature could discriminate benign from malignant disease.[br][bold]Conclusion:[/bold] It is feasible to collect cytologic material and ESS data real-time during ultrasound-guided FNAB using a miniaturized combined ESS-biopsy needle probe in the clinical setting. The cytologic specimen is adequate compared to conventional FNA biopsy, and the ESS data from this miniaturized optical probe is of comparable quality to the standard ESS probe used in the diagnosis of other malignancies. Preliminary analysis reveals a unique waveform signature that can differentiate benign from malignant thyroid nodules. In addition, the real time ESS data may allow improved targeting of biopsies and potentially reduce the numbers of insufficient biopsies. With the collection of further data, an algorithm using ESS collected using our novel miniaturized ESS biopsy probe could potentially be used as an [italic]in-situ[/italic] real time minimally invasive adjunct to conventional FNA cytology to immediately improve diagnosis and prevent unnecessary surgery.[br][br](1) Suh H et al., Annals Surg Oncol 2011;18(5):1300.[br][br]Sources of Research Support: Coulter Foundation Grant; BU-Fraunhofer Alliance for Medical Devices, Instrumentation and Diagnostics.[br][br]Nothing to Disclose: JER, HS, IDG, ER-D, OA, IJB, SLL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1506 302 2184 MON-277 PO43-01 Monday 1853 2012


1849 ENDO12L_MON-278 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Cytokine Modulation of Perlecan/HSPG2 Expression in Desmoplastic Stromal Cells and Prostate Cancer Cell Lines Curtis Robert Warren, Daniel D Carson, Mary C Farach-Carson Rice University, Houston, TX Perlecan/HSPG2 is the most abundant heparan sulfate (HS) proteoglycan in bone marrow stroma, where prostate cancer (PCa) cells find a metastatic niche in disease progression, but is typically low in normal stroma of glandular tissue such as the prostate. HS is essential to growth factor signaling in PCa cell growth and angiogenesis, and Perlecan expression increases locally in PCa-associated fibroblasts of the prostate after wounding or when cancer is present. The Perlecan protein and associated HS chains play a key role in tumor growth processes such as growth factor signaling and angiogenesis. We hypothesized that the desmoplastic response that increases perlecan expression in local stroma during PCa growth is driven by similar cell signaling processes in both the bone marrow and the prostate gland. To test this, we studied perlecan regulation in PCa cell lines undergoing epithelial-mesenchymal transdifferentiation, as well as bone marrow stromal and prostate stromal cell lines. An [italic]in silico[/italic] analysis of conserved transcription factor binding sites in the proximal promoter region of the perlecan gene led to the hypothesis that growth factors and cytokines present in the tumor microenvironment activate transcription factors leading to perlecan promoter activation. Using the 2.6 kb human perlecan proximal promoter region to drive a luciferase reporter, we found activation of the perlecan promoter by inflammatory cytokines in various PCa and stromal cell lines. Deletion analysis of the proximal promoter region in reporter assays demonstrated that the cytokine-responsive cis-element is encoded in the distal portion of the 2.6 kb promoter region, where two conserved putative NF[kappa]B binding sites reside. Real-time RT-PCR and dot-blot based analysis of mRNA and protein extracts, respectively, confirmed an increase in steady state levels of both transcript and protein after 72 hours of cytokine treatment. Ongoing studies include mutation of the perlecan promoter construct to identify the active cis element(s) responding to cytokine treatment and further characterization of the intracellular mechanism of perlecan transcriptional activation in cell lines and primary bone marrow stromal cells. Disruption of paracrine signaling interactions between PCa cells and surrounding stroma that increase perlecan expression during desmoplasia is expected to limit the stromal response and inhibit disease progression toward lethality.[br][br]Sources of Research Support: NIH/NCI P01 CA098912.[br][br]Nothing to Disclose: CRW, DDC, MCF-C 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2057 302 2185 MON-278 PO43-01 Monday 1854 2012


1850 ENDO12L_MON-279 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - I (1:30 PM-3:30 PM) Assay-Specific Effects of Age, Hypertension and Diabetes on Late-Night Salivary Cortisol in Overweight and Obese Individuals Susmeeta T Sharma, Smita B Abraham, Lynnette K Nieman Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD [bold]Background: [/bold]Late night salivary cortisol (LNSC) has been proposed as one of the screening tests for the diagnosis of Cushing[apos]s syndrome (CS). Abnormal LNSC results have been reported in elderly men with hypertension (HTN) and diabetes (DM) without a known diagnosis of CS. However, reference ranges are based on data from young, lean and healthy individuals and normative data in other populations are lacking.[br][bold]Objective: [/bold]To evaluate the effects of age [gt]55 years and presence of DM or HTN on the likelihood of having an abnormal Esoterix Radioimmunoassay (RIA) or Mayo tandem mass spectrometry (LC-MS/MS) LNSC.[br][bold]Methods: [/bold]Retrospective analysis of LNSC results from a prior cross-sectional prospective study that evaluated the specificity of screening tests for CS in overweight and obese subjects (1). Chi-square test was used to analyze difference between proportions.[br][bold]Results: [/bold]None of the 387 subjects (112 men) with available LNSC data had CS on follow-up. LNSC results were available for the RIA assay in 254 subjects, LC-MS/MS assay in 290 subjects, and both in 157 subjects. Forty (15.7%; 11 men) subjects had an abnormal RIA result while 22/290 (7.6%; 4 men) subjects had an abnormal result by LC-MS/MS. Compared to younger individuals, older subjects aged [gt]55 years were more likely to have an abnormal result by RIA (18.3% vs. 14.5%, P=0.4420) and LC-MS/MS (10.8% vs. 5.9%, P=0.1298) but these trends were not statistically significant. The likelihood of an abnormal RIA result increased with the presence vs absence of HTN (20.8% vs. 8.7%, P=0.0091) and DM (23.2% vs. 13.0%, P=0.0468). The presence of all co-morbidities (age [gt]55 years, HTN and DM) further increased the probability of an abnormal RIA result (11/28=39.3%) compared to subjects without this combination (29/226=12.8%) (P=0.0003). By contrast, there was no significant difference in the proportion of abnormal LC-MS/MS results in subjects with or without DM (5.3% vs. 8.4%, P=0.3733) or HTN (7.9% vs. 7.3%, P=0.8476) or the combination of co-morbidities (age [gt]55 years, DM and HTN, 12.5% vs. 7.6%; P=0.2755).[br][bold]Conclusion: [/bold]LC-MS/MS measurement of LNSC may be more accurate than RIA in elderly patients with DM and HTN, leading to less false positive results. Studies are needed to define the appropriate assay-specific diagnostic thresholds for LNSC in this population.[br][br](1) Baid SK et al., J Clin Endocrinol Metab 2009; 94:3857-3864.[br][br]Sources of Research Support: This work was funded in part by the intramural program of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health.[br][br]Nothing to Disclose: STS, SBA, LKN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1840 302 2186 MON-279 PO43-01 Monday 1855 2012


1851 ENDO12L_MON-280 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Characterization of Tumor Growth and Metastasis of Seven Unique Thyroid Cancer Cell Lines in Two [italic]In Vivo[/italic] Mouse Models of Advanced Thyroid Cancer Gregory Lund, Laura Pike, Xia Jing, Kevin Bauerle, Bryan Haugen, Rebecca Schweppe University of Colorado Anschutz Medical Campus, Aurora, CO Patients with advanced and metastatic thyroid cancer have a poor prognosis, especially those with bone metastases. Our aim was to develop and characterize more clinically relevant in vivo thyroid cancer models that will better allow for the study of advanced thyroid cancer. We have characterized the behavior of 7 unique thyroid cancer cell lines with different oncogenic mutations in both an orthotopic and distant metastatic mouse model of advanced thyroid cancer. Four anaplastic (8505C -BRAF V600E, SW1736 -BRAF V600E, C643 -NRAS Q61K, HTh74 -no known mutations) and three papillary thyroid cancer cell lines (BCPAP -BRAF V600E, TPC1 -RET/PTC1, K1 -BRAF V600E/PIK3CA E542K) were used to determine tumor establishment and growth in the orthotopic and metastatic models. All cell lines were engineered to express luciferase in order to monitor tumor growth and metastases in real time using bioluminescence imaging. In the orthotopic model, 5x10[sup]5[/sup] thyroid cancer cells are injected directly into the right thyroid lobe of an athymic nude mouse. Intrathyroidal tumor growth was robust in the cell lines 8505C (12/12 mice), BPCAP (11/12), HTh74 (9/9) and K1 (10/10). The remaining cell lines showed low or no growth in this model: SW1736 (0/8), C643 (1/8) and TPC1 (0/8). One limitation of the orthotopic model is that it does not produce consistent distant metastases. We therefore, also developed an intracardiac injection model that leads to predictable distant metastases. Thyroid cancer cells (1x10[sup]5[/sup]) are injected into the left ventricle of an athymic nude mouse. We have now shown that this model results in the development of clinically relevant metastases in 4/7 cell lines (8505C, BCPAP, C643, HTh74) with varying frequency based on cell type. 8505C and HTh74 cells produce metastatic tumors in nearly all of the mice. 8505C tumors form most readily in the bone but also form in the brain and soft tissue. HTh74 tumors form predominantly in the lungs and bone. BCPAP tumors variably formed (70% of mice) in the brain, bone, and soft tissue. Only 1/7 mice injected with C643 cells formed a distant spine metastasis. In summary, we have developed a novel metastatic in vivo model and characterized the growth of 7 different anaplastic and papillary thyroid cancer cell lines in two clinically relevant in vivo mouse models, providing new preclinical models to study novel therapies in advanced thyroid cancer.[br][br]Nothing to Disclose: GL, LP, XJ, KB, BH, RS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2015 303 2187 MON-280 PO43-02 Monday 1856 2012


1852 ENDO12L_MON-281 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) SRC-1 Potentiates Tumor Angiogenesis through Up-Regulation of HIF1a -Mediated VEGFa Expression Li Qin, Yixiang Xu, Zhen Feng, Yelin Wu, Yan Xu, Lan Liao, Jianming Xu Baylor College of Medicine, Houston, TX Steroid receptor coactivator 1 (SRC-1) belongs to the p160 coactivator family which also contains SRC-2 (TIF2) and SRC-3 (AIB1/pCIP/ACTR/RAC3) (1-3). These coactivators interact with nuclear receptors and other transcription factors to up-regulate gene expression (1-8). Accumulated evidence indicates that SRC-1 implicates in breast cancer progression and metastasis by promoting transcription of multiple genes important for tumor cell epithelial-mesencymal transition (EMT), migration, invasion and endocrine therapy resistance (6, 9-17). However, its role in tumor angiogenesis has not been examined. In our recent studies using SRC-1[sup]-/-[/sup]/PyMT and WT/PyMT mammary tumor cell lines established in our lab, we found that injection of SRC-1[sup]-/-[/sup]/PyMT cells to matrigel plug formed under the back skin in SCID mice induced less blood vessel formation compared with that of WT/PyMT tumor cells. Meanwhile, VEGFa mRNA level is significantly reduced in mammary tumors from SRC-1[sup]-/-[/sup]/PyMT transgenic mice. On the contrary, in SRC-1-overexpressed mammary tumors from MMTV/TVA/PyMT mice, expression of VEGFa dramatically increased. In agreement with these findings, in vitro studies revealed that VEGFa mRNA levels are much lower in two of the SRC-1[sup]-/-[/sup]/PyMT tumor cell lines compared with WT/PyMT cell lines. Knockdown of SRC-1 in WT/PyMT cells reduced VEGFa levels, while adenovirus-mediated re-expression of SRC-1 in SRC-1[sup]-/-[/sup]/PyMT cells significantly potentiated VEGFa expression. Furthermore, promoter analyze showed that SRC-1 interacts with HIF-1a to potentiate VEGFa promoter activities. We also found that both SRC-1 and HIF-1a are associated with the VEGFa promoter region by ChIP assays. These results indicate that SRC-1 can promote breast cancer progression and metastasis through enhancing tumor angiogenesis. Targeting SRC-1 may be a feasible strategy for inhibiting breast tumor angiogenesis and metastasis.[br][br]1. Onate SA, et al. Science 270(5240):1354-7. 2. Torchia J, et al. Nature 1997;387:677-84. 3. Xu J et al. Mol Endocrinol 2003;17:1681-92. 4. Kim HJ, et al. J Biol Chem 273(44):28564-7. 5. Xu J et al. Science 1998;279:1922-5. 6. Xu J, et al. Nat Rev Cancer. 9(9):615-30. 7. Na SY, et al. J Biol Chem 1998;273:10831-4. 8. Lee SK, et al. J Biol Chem 1998;273:16651-4. 9. Wang S, et al. Proc Natl Acad Sci U S A 2009;106:151-6. 10. Qin L, et al. Cancer Res 2009;69:3819-27. 11. Qin L, et al. Cancer Res 2011;71:1742-51. 12. Fleming FJ, et al. J Clin Pathol 2004;57:1069-74. 13. Myers E, et al. Br J Cancer 2004;91:1687-93. 14. Han JS, et al. BMC Cancer;10:629. 15. Redmond AM, et al. Clin Cancer Res 2009;15:2098-106. 16. McIlroy M, et al. Cancer Res;2010,70:1585-94. 17. McCartan D, et al. Cancer Res. 2012,72(1):220-9.[br][br]Sources of Research Support: NIH Grant CA112403, CA119689 and DK58242 to J. Xu, American Cancer Society Scholar Award (RSG-05-082-01-TBE) to J. Xu.[br][br]Nothing to Disclose: LQ, YX, ZF, YW, YX, LL, JX 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2041 303 2188 MON-281 PO43-02 Monday 1857 2012


1853 ENDO12L_MON-282 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) ER[beta]1 Represses Breast Cancer EMT by Destabilizing EGFR Christoforos Thomas, Gayani Rajapaksa, Fotis Nikolos, Ruixin Hao, Anne Katchy, Maria Bondesson, Savitri Krishnamurthy, Francisco Esteva, Jan-Ake Gustafsson University of Houston, Houston, TX; The University of Texas MD Anderson Cancer Center, Houston, TX Although epithelial to mesenchymal transition (EMT) is associated with specific breast cancer phenotypes, such as the basal-like and claudin-low tumors, there is a lack of validated markers, which can distinguish tumors with poor clinical outcome and high metastatic potential. Estrogen receptor beta (ER[beta]) has been associated with better survival in triple-negative breast cancers. However, it is still unclear if ER[beta] by influencing invasion can predict metastasis in breast cancer. Here we show that wild-type ER[beta] (ER[beta]1) inhibits EMT and invasion in basal cells when they grow either in vitro or in vivo in zebrafish. This correlates with an ER[beta]1-mediated upregulation of miR-200a-b-429 and the subsequent repression ZEB1 and SIP1, which results in increased expression of E-cadherin. Downregulation through ubiquitylation and degradation of the basal marker EGFR was found to be involved in the ER[beta]1-mediated repression of EMT since induction of EGFR expression or treatment with EGF abolished the ability of ER[beta]1 to sustain the epithelial phenotype. This inverse correlation of ER[beta]1 with EGFR and its positive association with epithelial markers was additionally observed in breast cancer specimens. These results strengthen the association of ER[beta]1 with the regulation of EMT and propose the receptor as potential crucial marker in predicting survival and metastasis in breast cancer.[br][br]Nothing to Disclose: CT, GR, FN, RH, AK, MB, SK, FE, J-AG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2104 303 2189 MON-282 PO43-02 Monday 1858 2012


1854 ENDO12L_MON-283 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Molecular Profiling of Advanced Thyroid Carcinomas Naifa L Busaidy, Nadia N Megahed, Maria E Cabanillas, Katherine S Hale, Camilo Jimenez, Mouhammed A Habra, Michael Kupferman, Elizabeth Grubbs, Gordon Mills, Michelle D Williams University of Texas - MD Anderson Cancer Center, Houston, TX; University of Texas - MD Anderson Cancer Center, Houston, TX; University of Texas - MD Anderson Cancer Center, Houston, TX; University of Texas - MD Anderson Cancer Center, Houston, TX; University of Texas - MD Anderson Cancer Center, Houston, TX [bold][underline]Introduction:[/underline][/bold] Local regional recurrence and distant metastases occur in up to 30% of thyroid cancer patients. Although some tumors remain avid for radioactive iodine, alternative therapies are needed for progressive patients. Molecular profiling of thyroid tumors for alterations known in other tumor types may allow for the detection of directed targets for therapy.[br][bold][underline]Methods[/underline]:[/bold] Tumors of follicular origin were obtained from our tumor bank. DNA was extracted from formalin-fixed paraffin embedded tumor tissue and was analyzed for 145 mutations/alterations in 30 genes by mass spectrometry based DNA analysis (Sequenom Inc, San Diego, CA).[br][bold][underline]Results[/underline][/bold]: We analyzed tumor tissue from 51 patients with advanced or recurrent thyroid carcinomas of follicular origin. Tumor tissue tested included 32 (62%) primary tumors, 15 (29%) locoregional recurrences, and 5 (10%) from distant metastatic sites. Two of the 30 genes analyzed showed mutations including an nRAS (Q61R) mutation in 1 of 3 follicular/hurthle cell tumors and BRAF (V600E) in 18 of 30 (60%) papillary thyroid carcinomas, 1 of 7 (14%) poorly differentiated and 1 of 6 (17%) anaplastic thyroid carcinomas. By sites tested, BRAF V600E was present in 67% primary tumors, 70% of the recurrences, but only 20% of the distant metastatic sites. Interestingly, 33% of PTC patients younger than 45 years at diagnosis had a positive BRAF mutation versus 68% of those 45 years or older. BRAF mutation was not associated with lower RAI avidity (p=1.0), nor with lower survival (p=0.9).[br][bold][underline]Conclusion: [/underline][/bold]Advanced thyroid cancers harbor prevalent BRAF mutations beginning in tumorigenesis. However, BRAF mutations alone do not differentiate outcomes in these advanced thyroid carcinoma patients, even though older patients were two times more likely to have a BRAF mutation and to be RAI nonavid. Moreover, the lower rate of BRAF mutation in distant metastatic disease and poorly differentiated tumors emphasizes the need for continued tumor analysis for yet identified tumorigenic factors.[br][br]Sources of Research Support: Kleberg Center for Molecular Markers at The University of Texas - M.D. Anderson Cancer Center.[br][br]Nothing to Disclose: NLB, NNM, MEC, KSH, CJ, MAH, MK, EG, GM, MDW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2284 303 2190 MON-283 PO43-02 Monday 1859 2012


1855 ENDO12L_MON-284 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Higher TSH Level Is the Risk Factor for Differentiated Thyroid Carcinoma (A Large Population-Based Study) Hee Kyung Kim, Jee Hee Yoon, Soo Jung Kim, Ho-Cheol Kang Chonnam National University Medical School, Gwangju, Korea [bold]BACKGROUND: [/bold]Higher TSH level has been reported to be associated with differentiated thyroid cancer (DTC). To validate the association, we compared the TSH levels obtained from patients with DTC and those from general population whose TSH level were within normal range.[br][bold]METHODS:[/bold] The case group includes 1,759 patients with DTC who underwent thyroid surgery at Chonnam National University Hwasun Hospital. General population (n=1,548) who participated the Thyroid Disease Prevalence Study was used as a healthy control group. A total of subjects were divided into four groups of similar size according to their TSH levels, with the first quartile used as the reference.[br][bold]RESULTS:[/bold] The mean TSH level of the case group was significantly higher than control group (1.95 [plusmn] 0.9 mIU/L vs. 1.62 [plusmn] 0.8 mIU/L, [italic]p[/italic] [lt] 0.001), and it was significantly associated with DTC risk. The multiple logistic regression, after controlling of age, sex, family history of thyroid cancer, showed that the odds ratios and 95% confidence interval for increasing quartiles of TSH level were 1.27 (1.03-1.57), 1.55 (1.25-1.92) and 2.21 (1.78-2.74), respectively. No significant differences were observed in mean TSH levels between tumor stages and tumor sizes.[br][bold]CONCLUSION:[/bold] Higher TSH level within normal range is the independent risk factor for DTC, and it may contribute to the initiation step of thyroid carcinogenesis. TSH levels in patients with thyroid nodules may be used as the diagnostic adjunct in the identification of high-risk patients who require further investigation and/or surgical intervention.[br][br]Nothing to Disclose: HKK, JHY, SJK, H-CK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 506 303 2191 MON-284 PO43-02 Monday 1860 2012


1856 ENDO12L_MON-285 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) IGF-I Signaling Enhances Levels of Choline Kinase in Breast Cancer Cells: Implications for Developing Noninvasive Biomarker of Response to IGF1R Targeted Therapy Huy T Donguyen, Deepali Sachdev, Joseph C Weber University of Minnesota, Minneapolis, MN; University of Minnesota, Minneapolis, MN Insulin-like growth factor I (IGF-I), a peptide growth factor, signals through the type I insulin-like growth factor receptor (IGF1R) which is a transmembrane receptor tyrosine kinase. Multiple drugs targeting IGF1R are being tested in clinical trials. Biomarkers that enable the selection of patients with IGF driven tumors or the monitoring of response to this therapy have not been yet been identified. Previously, we found that total choline (tCho) metabolites decreased in MCF-7 xenograft tumors in mice twenty-four hours following administration of an inhibitory antibody against IGF1R. The tCho was measured by [sup]1[/sup]H magnetic resonance spectroscopy (MRS) and included three species of choline containing compounds, choline, phosphocholine (pCho), and glycerophosphocholine. This suggests that noninvasive measurement of tCho may be a useful measure of response to IGF1R targeted therapy. The objective of the current study is to investigate whether there is a mechanistic link between IGF signaling and choline phospholipid metabolism to understand if decreased tCho levels could be a specific marker of inhibition of IGF signaling in cancer cells.[br]In this study, we examined the effect of IGF signaling on levels of choline kianse (CK). Choline kinase (ChoK) generates pCho, one of the species measured by MRS in our prior in vivo study. MCF-7, MDA-MB-231 and MDA-MB-435A/LCC6 breast cancer cells were treated with IGF-I for 10 minutes, 4h or 24h and levels of ChoK were examined by western blotting. IGF-I signaling upregulated ChoK levels at 24h in MCF-7 cells but not in MDA-MB-231 and MDA-MB-435A/LCC6. In MCF-7 cells, IGF signaling activates the mitogen-activated protein kinase (MAPK). However, in MDA-MB-231 and MDA-MB435A/LCC6 cells phosphorylation of p44/p42 MAPK are unaffected by IGF-I signaling or IGF1R inhibition. These data suggest that IGF-I induced upregulation of ChoK may be via a MAPK dependent mechanism. We next investigated the effect of AVE1642, an inhibitory antibody against IGF1R, on IGF-I stimulated upregulation of ChoK. IGF-I induced upregulation of ChoK was blocked byAVE1642.[br]These results provide molecular evidence for the observed decrease of tCho after IGF1R inhibition in xenograft tumors. This study is clinically significant as it provides evidence for developing decreased tCho, following IGF1R targeted drugs, as a specific biomarker for inhibition of IGF signaling.[br][br]Sources of Research Support: This work was supported by ACS grant IRG-58-001-49 IRG#6 (D.S) and a philanthropic gift from the Atwaters. We thank Immunogen for providing AVE1642 and isotype matched control antibody.[br][br]Nothing to Disclose: HTD, DS, JCW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1785 303 2192 MON-285 PO43-02 Monday 1861 2012


1857 ENDO12L_MON-286 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) High Expression of Claudin-1, HBME-1, p16 and CK19 in Papillary Thyroid Carcinoma Mi-Kyung Kim, Hye-Soon Kim, Sang-Yoon Kim, Ho Chan Cho, Sun-Young Kwon, Eui Dal Jung Keimyung University School of Medicine, Daegu, Republic of Korea; Keimyung University School of Medicine, Daegu, Republic of Korea; Catholic University of Daegu, School of Medicine, Daegu, Republic of Korea Disruption of the cell-to-cell junction with changes in the expression of the junctional proteins is the hallmark of cancer invasion and metastasis. The claudins, a family of transmembrane proteins associated with tight junctions, are critical for maintaining cell-to-cell adhesion in sheets of epithelial cells and differentially expressed in malignant tumors as compared to the corresponding healthy tissues. Altered expression of claudin-1 has been reported in several tumor types, including endometrial, papillary renal cell and colonic carcinoma, and increased claudin-1 mRNA levels have been observed in papillary thyroid carcinoma (PTC). However, we do not know the role of claudin-1 in the carcinogenesis and progression of PTC. Therefore, in this study, we investigated the expression patterns of cluadin-1 between PTC and benign nodule and the relationship with other biomarkers HBME-1, p16, CK19 in Korean. This study included 239 cases with thyroid nodules who were resected surgically at Keimyung University Dongsan Medical Center from 2002 to 2011. 207 patients had PTC and 32 patiens had benign nodule. We examined claudin-1, HBME-1, p16, CK19 and [beta]-catenin expression using immunohistochemical (IH) staining. The IH scores of claudin-1, HBME-1, p16, CK19 showed a significant difference between PTC and benign thyroid nodule (p [lt] 0.0001 respectively). But the IH score of [beta]-catenin did not show a significant difference between two groups (p = 0.842). In addition, the IH scores of claudin-1, HBME-1, CK19, p16 and [beta]-catenin were not associated with vascular invasion, regional lymph node metastasis, multicentricity, extrathyroidal extension, distant metastasis.[br]In conclusion, these results suggest that higher expressions of claudin-1, HBME-1, p16 and CK19 were found in PTC than benign thyroid nodule. Therefore, we suggest they may represent novel tumor markers for PTC.[br][br]Nothing to Disclose: M-KK, H-SK, S-YK, HCC, S-YK, EDJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 494 303 2193 MON-286 PO43-02 Monday 1862 2012


1858 ENDO12L_MON-287 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Multiplicity as a Prognostic Factor of Papillary Thyroid Carcinoma Jung Min Kim, Jong Chul Won, Kyung Soo Ko, Byoung Doo Rhee Sanggye Paik Hospital, Inje University College of Medicine, Seoul, Korea Backgrounds: Mutiplicity of PTC are not an unusual finding, although the origin of these foci is unclear. Either intraglandular metastases from a single dominant tumor or unrelated neoplastic clones were definitively proven as the means by which multicentric PTC form. In addition, there is insufficient clinical information concerning multicentric PTC presentation, prognosis, and long-term follow-up studies after treatment. Multiplicity of papillary thyroid carcinoma (PTC) has not been considered as an independent prognostic factor from a variety of tumor staging systems.[br]Aims: To evaluate whether that the presence of mulciplicity would be associated with tumor recurrence in PTC patients.[br]Methods: A total 249 PTC patients at a single institution who underwent total thyroidectomy and node dissection were retrospectively reviewed; the mean follow-up period was 2.8 years. Postoperative radioactive iodide ablation for thyroid remnant was performed after surgery for most patients.[br]Results: Of all the PTC cases reviewed, 85 cases (34%) were categorized as multicentric PTC. Compared with patients with unifocal PTC, multicentric PTC patients demonstrated higher cervical lymph node metastasis and tumor recurrence. Multiplicity was also significantly associated with tumor recurrence; 6% vs. 1% with and without multiplicity, respectively (P = 0.022 by log-rank test). However, this association was lost on multivariate analysis adjusting for conventional clinicopathological predictors of recurrence.[br][br]Nothing to Disclose: JMK, JCW, KSK, BDR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 56 303 2194 MON-287 PO43-02 Monday 1863 2012


1859 ENDO12L_MON-288 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Prolactin Gene Expression in Primary Central Nervous System Tumors Graziella Alebrant Mendes, Julia FS Pereira-Lima, Maria Beatriz Kohek, Geraldine Trott, Marlise Di Domenico, Carolina Garcia Soares Leaes, Nelson Pires Ferreira, Miriam da Costa Oliveira Universidade Federal de Ci[ecirc]ncias da Sa[uacute]de de Porto Alegre, Porto Alegre, Brazil Introduction: Prolactin (PRL) is a hormone synthesized in both the pituitary gland and extrapituitary sites. It has been associated with the occurrence of neoplasms and, more recently, with central nervous system neoplasms. The aim of this study was to evaluate prolactin expression in primary central nervous system tumors through quantitative real-time PCR and immunohistochemistry. Materials and methods: Cross-sectional study of a sample of 70 patients with anatomopathological diagnosis of primary central nervous system tumor who underwent neurosurgery. This study was approved by the Research Ethics Committee of the Federal University of Health Sciences of Porto Alegre (UFCSPA), all patients signed an informed consent form, and the authors signed a term of confidentiality. After surgery, small tumor samples were quickly frozen in liquid nitrogen and stored in freezer at -80[deg]C for subsequent prolactin evaluation through quantitative real-time PCR and the remaining material was fixed in formalin and embedded in paraffin for prolactin assessment by immunohistochemistry. Results: Patient mean age was 49.1 years (SD 15.43), and females accounted for 70% of the sample. The most frequent subtype of histological tumor was meningioma (61.5%), followed by glioblastoma (22.9%). Twenty cases (28.6%) showed prolactin expression by immunohistochemistry, most of them females (18 cases, 90%). Statistical significance was obtained by relating gender (18 females versus 2 males) to immunohistochemistry PRL positivity (p = 0.043). Quantitative real-time PCR did not show any prolactin expression. Conclusions: There is no data in the current literature about PRL gene expression by quantitative real-time PCR in normal samples of the CNS and there are no studies analyzing PRL gene expression in different types of CNS tumors either by conventional PCR or by quantitative real-time PCR. Despite the presence of prolactin expression by immunohistochemistry, the lack of its expression by quantitative real-time PCR indicates that its presence in primary tumors in central nervous system is not a reflex of local production. Thus, further studies are needed regarding the origin of this PRL identified with the immunohistochemistry technique and its true role in the pathogenesis of central nervous system tumors.[br][br]Nothing to Disclose: GAM, JFSP-L, MBK, GT, MDD, CGSL, NPF, MdCO 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 67 303 2195 MON-288 PO43-02 Monday 1864 2012


1860 ENDO12L_MON-289 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Association of Decreased Insulin Sensitivity and Hyperinsulinemia with Increased Risk of Urinary Bladder, Cervix, Lung, Head and Neck Cancers S K Singh, A Gahlot, M Pandey, S Pradhan Institute of Medical Sciences, Varanasi, India; Institute of Medical Sciences, Varanasi, India; Institute of Medical Sciences, Varanasi, India [bold]Introduction:[/bold] Studies focussing causal relationship between hyperinsulinemia, insulin resistance with cancer is limited and variable. Few studies have shown association of dysglycemia, hyperinsulinemia with cancers of gastrointestinal tract, breast, prostate and endometrium. Present study was designed to study association of hyperinsulinemia, insulin sensitivity and dysglycemia with hitherto unreported urinary bladder, cervix, lung, head and neck cancers.[br][bold]Methods:[/bold] We analyzed 100 patients with cancers involving urinary bladder, cervix, lung, head and neck cancers and 100 healthy controls. After detailed clinical examination, fasting and 2 hour post oral (75 gm) glucose, concentrations of serum glucose, plasma insulin and C-peptide were measured. Insulin resistance/sensitivity by homeostatic model assessment (HOMA-IR) and quantitative insulin sensitivity check index (QUICKI), Insulin Sensitivity Index (ISI-Gutt) was calculated using mathematical tools. The statistical analysis was done using software SPSS for windows (Version 16). Multivariate logistic regression models estimated age and BMI adjusted relative risks as odds ratios (ORs).[br][bold]Results:[/bold] Mean fasting insulin, fasting C-peptide and HOMA-IR was 20% (p=0.037), 73% (p=0.001), 24% (p=0.025) higher in cancer group respectively. Mean QUICKI was 10% (p[lt]0.001) and ISI-Gutt was 28% (p[lt]0.001) lower in cancer group in comparison to controls. Among subjects with higher C-peptide tertiles (second and third), compared with lowest tertile (first), there was significantly increased risks of cancer with OR of 2.55 (95% confidence interval [CI] = 1.13-5.76, P=0.023); and 3.54 (95% CI = 1.62-7.71, P=0.001); respectively. A similar pattern was observed with HOMA-IR (OR = 2.57, 95% CI = 1.07- 6.13; P =.03) for highest vs. lowest tertile. The cancer risk was significantly higher in lowest tertile (third) of QUICKI and ISI-Gutt as compared to highest tertile (first) with OR of 2.40 (95% CI = 1.21- 5.78; P =0.04) and 5.80 (95% CI = 2.46- 13.67; P [lt]0.001) respectively.[br][bold]Conclusion:[/bold] Elevated fasting plasma insulin and C-peptide was associated with a higher risk of urinary bladder, cervix, lung, head and neck cancers. Of various indices of insulin resistance and sensitivity, whole body insulin sensitivity index was most significantly associated with cancer risk in a reciprocal manner.[br][br]Nothing to Disclose: SKS, AG, MP, SP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 968 303 2196 MON-289 PO43-02 Monday 1865 2012


1861 ENDO12L_MON-290 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Estrogen Mediates Pro-Tumorigenic Activity of Prostate Carcinoma-Associated Fibroblasts and Mast Cells Via CXCL12 Renea A Taylor, Stuart J Ellem, Luc Furic, Ola Larsson, Mark Frydenberg, John Pedersen, Grant Buchanan, Eleanor Need, Andrew Trotta, Gail P Risbridger Monash University, Clayton, Australia; Monash University, Clayton, Australia; Basil Hetzel Institute, Adelaide, Australia; Karolinska Institutet, Stockholm, Sweden Prostate tumour stroma plays a major role in carcinogenesis and recent studies show a consistent association between loss of stromal androgen receptor (AR) and poor clinical outcome in patients. Significantly, there is a direct link between AR and estrogen receptor [alpha] (ER[alpha]) expression, with ER[alpha] promoting adverse effects in the prostate. This study examined stromal ER[alpha] within the tumor microenvironment.[br]Gene microarrays of patient-matched primary carcinoma-associated fibroblasts (CAFs) and normal prostatic fibroblasts (NPFs) showed loss of AR with an elevation of ER[alpha] expression (ER[beta] and PR levels were low/unchanged). This imbalance of AR/ER[alpha] expression in tumor stroma was confirmed by immunohistochemistry on radical prostatectomy specimens. To assess steroid receptor function, cells were transfected with AR-targeted and/or ER-targeted luciferase reporters, which revealed that overall AR responsiveness was markedly diminished in CAFs compared to NPFs, whereas the ER[alpha] response was maintained. Thus, in the tumor microenvironment, there is a shift to estrogen/ER[alpha] dominance. To assess the molecular consequences of this, we compared our CAF array data to publically available estrogenic gene signatures, identifying CXCL12 as a major estrogen driven target gene in CAFs (aka SDF-1; a multi-potent chemokine involved in inflammation, ECM degradation and angiogenesis). To investigate a role of CXCL12, we examined its influence on mast cells, having previously identified these cells in murine estrogen-induced prostate inflammation and malignant transformation. Mast cells were increased in the peritumoral region of prostate cancer in patient tissues, while, [italic]in vivo[/italic], mast cell migration was stimulated by CAF condition media, this being CXCL12 mediated (shown via recombinant CXCL12 and CXCR4 inhibitor, AMD3100). Mast cells also expressed ER[alpha], with estrogen directly stimulating mast cell proliferation and production of additional cytokines and chemokines such as CXCL5 and CCL20, reported as pro-tumorigenic in the prostate.[br]Overall, these data show that the prostatic tumor microenvironment is dominated by estrogen signalling (via ER[alpha] and the absence of AR). This stimulates specific cytokine and chemokine expression, including CXCL12, promoting the recruitment of mast cells to the tumor site, which further alter the local microenvironment under the influence of estrogen. Collectively, we present a potential mechanism for the pro-tumorigenic actions of prostatic tumor stroma.[br][br]Nothing to Disclose: RAT, SJE, LF, OL, MF, JP, GB, EN, AT, GPR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1358 303 2197 MON-290 PO43-02 Monday 1866 2012


1862 ENDO12L_MON-291 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) ER[beta] Inhibits Lung Cancer Cell Growth by Inducing Cell Cycle Arrest and Apoptosis Fotis Nikolos, Gayani Rajapaksa, Igor Bado, Christoforos Thomas, Jan-Ake Gustafsson University of Houston, Houston, TX Non-small cell lung cancer (NSCLC) is the leading cause of cancer deaths in the United States and worldwide. There is an urgent need for development of new therapeutic approaches and discovery of novel therapeutic targets. Recent studies have suggested that estrogens may play a role in stimulating NSCLC development and progression. Estrogen receptors, the mediators of biological effects of estrogens and anti-estrogens are very important for the regulation of cell proliferation and differentiation in various human tissues. The purpose of this study is to investigate the role of estrogen receptor beta (ER[beta]1) in NSCLC initiation and progression. We analyzed cell cycle progression, apoptosis and expression of cell cycle regulatory proteins in control and ER[beta]1-expressing lung cancer cells. Here we show that induction of ER[beta]1 expression inhibited growth in H1299 NSCLC cells. Treatment of these cells with 17[beta]-estradiol or 3[beta]-Adiol further decreased cell proliferation and induced apoptosis. ER[beta]1 was found to inhibit cell growth by altering the expression of cell cycle regulatory proteins, including cyclin-D1, cyclin-E, p27 and cdk2. These results suggest that, by regulating cell cycle and apoptosis, ER[beta]1 affects NSCLC cell growth. Thus, ER[beta]1 could be an important player in biology of lung cancer and a potentially useful component in the clinical management of the disease.[br][br]Nothing to Disclose: FN, GR, IB, CT, J-AG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1668 303 2198 MON-291 PO43-02 Monday 1867 2012


1863 ENDO12L_MON-292 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Thyroid Hormone-Responsive Protein Spot 14 (S14) Accelerates Mammary Tumor Formation and Enhances Cell Proliferation in MMTV-Neu Mice Elizabeth A Wellberg, Michael C Rudolph, Andrew Lewis, Steven M Anderson University of Colorado Anschutz Medical Campus, Aurora, CO Thyroid Hormone Responsive Protein Spot 14 (S14) plays a role in regulating enzymes involved in lipid synthesis in the liver, adipose tissue, and the lactating mammary gland. S14 null mice suffer a lactation defect characterized by a reduction in [italic]de novo[/italic] synthesized fatty acids (C[le]16) present in milk, which results in a reduced growth rate for suckling pups. The mechanism through which S14 influences fatty acid synthesis is not clear. In human breast tumors, high levels of fatty acid synthase (FASN) expression and activity are often associated with aggressive tumors. Additionally, high levels of S14 and FASN expression in breast tumors are associated with a poor clinical prognosis for women. Published data suggest a link between S14, FASN and breast cancer prognosis, yet the studies to date are correlative. We hypothesized that S14 enhances mammary tumor growth and metastasis by enhancing FASN activity and increasing de novo fatty acid synthesis in the breast epithelium. We have developed a mouse model to causatively link S14 expression and mammary tumor progression and metastasis. Transgenic mice were created that express S14 under control of the mouse mammary tumor virus (MMTV) promoter. These mice were crossed with the well-characterized MMT-Neu mammary tumor mouse model. MMTV-Neu/MMTV-S14 mice (Neu/S14) developed tumors in 237.5 days, which was significantly faster than control MMTV-Neu (Neu) mice, which developed tumors in 259 days in this study (p=0.011). Although differences in the activation of the ErbB2/Neu signaling pathway were not observed between groups in late-stage tumors, cell proliferation was higher in tumors from Neu/S14 mice compared to Neu controls. We also observed more branching and alveolar bud-like structures in non-tumor bearing glands of Neu/S14 mice that were not seen in the mammary glands of Neu controls, suggesting S14 influences early events in tumorigenesis. Studies are ongoing to characterize the aqueous and lipid metabolite profile of single and bi-transgenic tumors, and to determine if high S14 expression in mammary tumors correlates with increased lung metastasis.[br][br]Sources of Research Support: EAW is supported by DoD BCRP Fellowship BC098051; MCR is supported by DoD BCRP Fellowship BC810596.[br][br]Nothing to Disclose: EAW, MCR, AL, SMA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2022 303 2199 MON-292 PO43-02 Monday 1868 2012


1864 ENDO12L_MON-293 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Role of Monocyte Activation Markers in Patients with Localized and Metastatic Breast Cancer. Preliminary Results Franco Lumachi, Filippo Marino, Giordano B Chiara, Stefano MM Basso, Francesco Fallo University of Padova, School of Medicine, Padova, Italy; University of Padova, School of Medicine, Padova, Italy; S Maria degli Angeli Hospital, Pordenone, Italy; University of Padova, School of Medicine, Padova, Italy [bold]Background[/bold]: The decrease in use of hormone replacement therapy among postmenopausal women has been suggested as an explanation for the decrease in breast cancer (BC) incidence registered in recent years. However, a number of patients with small BC may develop aggressive disease, and thus the traditional histopathological characteristics are unable to correctly evaluate the biological behavior of BC. Each tumor is not an isolated entity, but is associated with several host cells, such as endothelial cells, fibroblasts, and immune cells. Monocyte activation plays a role in immune response against cancer, and contributes to cancer cells aggressiveness. The aim of this study was to assess the role of monocyte activation, showed using three monoclonal antibodies against human CD64 (clones 22 and 32.2) and CD163 (clone Mac2-158), in differentiating patients with localized and metastatic BC.[br][bold]Patients and Methods[/bold]: Sixteen women (median age 47 years, range 32-76 years) with confirmed invasive ductal carcinoma and positive axillary lymph nodes (pN1) who were not currently receiving chemotherapy (cases), and 18 age- and pT-matched pN0 women (controls) were enrolled in the study. The absolute neutrophil and monocyte count was obtained with routine hematological assay, while polymorphonuclear (PMN) CD64 index, monocyte CD64 index (as direct marker of monocyte activation), and monocyte CD163 index were assayed using a specific commercial kit. Each sample (50 [mu]L) was prepared and analyzed by flow cytometry, according to the manufacturer[apos]s guidelines.[br][bold]Results[/bold]: The absolute neutrophil and monocyte counts did not differ ([italic]p[/italic]=NS) between groups (cases [italic]vs[/italic]. controls), as well as PMC CD64 index (7.5[plusmn]2.3 [italic]vs.[/italic] 8.5[plusmn]2.1, [italic]p[/italic]=0.19). The monocyte CD64 index was significantly higher in patients with metastatic BC (10.5[plusmn]2.1 [italic]vs[/italic]. 8.4[plusmn]1.9, [italic]p[/italic]=0.004), while monocyte CD163 index was lower (2451[plusmn]256 [italic]vs[/italic]. 2711[plusmn]365, [italic]p[/italic]=0.024).[br][bold]Conclusions[/bold]: Usually, PMN CD64 index increases during infections in response to specific cytokines. An elevated monocyte CD64 index suggests that monocyte vs. neutrophil activation may have a role in mediating the response to the presence of metastases, while a lower monocyte CD163 index suggests a scavenger receptor shedding during monocyte activation. These preliminary data confirm the usefulness of monocyte activation markers measurement in patients with metastatic BC.[br][br]Goodale D et al., Cytometry B Clin Cytom 2009; 76: 107. Lumachi F et al., Cytokines 2010; 50: 229.[br][br]Nothing to Disclose: FL, FM, GBC, SMMB, FF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1214 303 2200 MON-293 PO43-02 Monday 1869 2012


1865 ENDO12L_MON-294 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) The Hormones BNP and FSH in Ovarian Cancer Larn Hwang, Kouros Motamed, Chao Hsiao, Vuong Trieu Biomiga Diagnostics, Fountain Valley, CA; Autotelic Lab, Fountain Valley, CA Background: CA125 is regarded as a serum tumor marker for monitoring response to chemotherapy, detecting disease recurrence, as well as distinguishing malignant from benign pelvic masses. However, it is not an appropriate diagnostic biomarker as the majority of healthy women with high levels of CA125 do not have cancer. This study evaluated CA125 along with the hormones FSH and BNP in relation to hypertension. Methods: Serum samples collected at time of diagnosis of ovarian cancer were tested using rapid and quantitative point-of-care (POC) devices for blood biomarkers (CA125, FSH, and BNP) and the data was evaluated using JMP9 statistical analysis software. Results: 41 ovarian cancer patients were analyzed for CA125 prior to surgery of which 3 (7%), 11 (27%), and 27 (66%) were clear cell carcinoma, cystadenocarcinoma and adenocarcinoma, respectively. The dominant histological subtype was serous (29 of 41, 71%), followed by endometrioid (5 of 41, 12%) and mucinous (4 of 41, 10%). Seventeen patients exhibited CA125 levels of less than 130 U/ml (low CA125 group) and 24 patients exhibited CA125 levels of greater than 130 U/ml (high CA125 group). The cutoff resulted in the clean separation of the mucinous subtype (5 of 5) into the high CA125 group and the endometrioid subtype (4 of 4) into the low CA125 group, p = 0.026, Pearson. The hypertensive group exhibited significantly higher incidence of high CA125 (17 of 20, 85%, vs. 7 of 21, 33%, p = 0.0008, Chi-square. In the serous adenocarcinoma group, FSH level was higher (median=151.6 mU/ml) vs. normal controls (median of 13.4 mU/ml, p = 0.01, Wilcoxon). Moreover, incidence of BNP [gt] 25 pg/ml was higher for patients (14 of 19, 74%) vs. normal controls (3 of 10, 30%, p = 0.02, Chi-square). FSH progressively increased from normal controls, to normotensive patients, to hypertensive patients with median FSH values of 13.4, 79.3, and 232.2, respectively. The same was not observed for BNP. These data suggest the BNP and FSH hormones play a role(s) in ovarian cancer and should be examined further. Conclusions: This small retrospective study suggested that hypertension modulates the CA125 level as well as the hormone FSH, but not BNP in ovarian cancer. Elevated FSH and BNP levels in ovarian cancer should be examined further as biomarkers for ovarian cancer. The POC device described here could facilitate this application by making biomarker testing more readily deployable during clinical trials.[br][br]Nothing to Disclose: LH, KM, CH, VT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1376 303 2201 MON-294 PO43-02 Monday 1870 2012


1866 ENDO12L_MON-295 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Comparison of Thyroglobulin Doubling Time with Classical Prognostic Factors in Patients with Differentiated Thyroid Cancer in Pakistan Population Asma Ahmed, Zain Bhutta, Saira Furqan, Safia Awan, Najmul Islam The Aga Khan University Hospital (AKUH), Karachi, Pakistan Background:[br]Several classical prognostic factors (age, gender [amp] TNM staging) have been proposed for papillary thyroid cancer (1). Recently, there had been some studies showing relationship of thyroglobulin doubling time with prognosis of thyroid cancer (2). We compared these different prognostic factors of thyroid cancer in our population.[br]Methods:[br]Records of patients with thyroid cancer attending Endocrine clinics of tertiary care hospital at Karachi, Pakistan between 1999-2011 were retrieved and analyzed retrospectively. Tg doubling time was computed using Tg values measured during routine follow-up. Patients were followed for a mean of 6.45 years and a median of 6 years. The mean age of the patients at the time of initial presentation was 39 years.[br]Results:[br]Of the 207 patients, 165 patients had differentiated carcinoma. Out of these, 71 patients had 4 or more values of stimulated thyroglobulin levels with negative antibody levels. These patients were grouped into two main categories of positive and negative doubling time (median -1.4300). Most of the patients 54 (76%) fell in to the category of negative doubling time in comparison to only 17 patients (24%) who had positive doubling time. Among all the prognostic factors TNM staging was found to be better predictor of distant metastasis (p=0.001) and persistence of disease (p=0.004). Age[ge]50 years was found to be associated with distant metastasis (p=0.007) and recurrence of the disease (p=0.057). Other factors like gender and thyroglobulin doubling time did not show any association with overall cure, persistence and recurrence rate (p=NS).[br]Conclusion:[br]TNM staging still remains strong prognostic factor in Pakistani population with differentiated thyroid cancer. Age cut off of 50 years instead of 45 was found to be associated with distant metastasis and recurrence of disease. Interstengly, our study did not show any association of thyroglobulin time with prognosis of thyroid carcinoma.[br][br](1)Cooper et al., Revised American Thyroid Association management guidelines for patients with thyroid nodules and differentiated thyroid cancer.Thyroid 19:1167-1214. (2)Akira et al.,Prognostic impact of serum thyroglobulin doubling time under thyrotropin suppression in patients with papillary thyroid carcinoma who underwent total thyroidectomy. Thyroid 21;2011.[br][br]Nothing to Disclose: AA, ZB, SF, SA, NI 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1585 303 2202 MON-295 PO43-02 Monday 1871 2012


1867 ENDO12L_MON-296 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) SDHB-Related Metastatic Pheochromocytomas/Paragangliomas (SDHB-MET) Lack a Therapeutic Target for Tyrosine Kinase Inhibitors Joey Matro, Marsha Merrill, Hana Turkova, Thanh-Truc Huynh, Uma Shankavaram, Stefan Grebe, Electron Kebebew, Maria Merino, Antonio Tito Fojo, Karel Pacak National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; Mayo Clinic, Rochester, MN; University of the Philippines-Diliman, Quezon City, Philippines; University of Santo Tomas, Manila, Philippines Pheochromocytomas (PHEOs)/paragangliomas (PGLs) are rare tumors of the chromaffin cells. Ten genes have been identified in familial cases and of these, mutations in succinate dehydrogenase B (SDHB) are the most aggressive. No satisfactory treatment is currently available and prognosis remains poor. Although useful in other malignancies, the tyrosine kinase inhibitor sunitinib has not appeared very effective based on our clinical observations at our institution, and published case reports have been controversial. Sunitinib targets a number of receptor tyrosine kinases, of which vascular endothelial growth factor receptor 2 (VEGFR2) is thought to be the most important because its inhibition may lead to decreased angiogenesis that might in turn impair tumor growth.[br]The purpose of this study was to investigate possible mechanisms for the lack of efficacy of sunitinib in metastatic SDHB-related PHEOs/PGLs (SDHB-MET). We performed RT-PCR of target receptors including VEGFR-1, -2, and -3; platelet derived growth factor (PDGFR) A and B; colony stimulating factor 1 receptor (CSF1R) and v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog (KIT). Compared to normal human adrenal medulla (NAM), primary PHEOs/PGLs, including those with SDHB mutations, had higher levels of expression of the various sunitinib targets. In VEGFR2, for example, fold changes of 1.56 to 10.97 relative to NAM were observed in the four different primary PHEO/PGL groups we tested. However, the SDHB-MET expression profile was similar to that found in NAM, with the exception of KIT, which was downregulated for all tumor types. VEGFR2 immunohistochemistry (IHC) showed a similar result, indicating a lack of upregulation of VEGFR2 in the vasculature of SDHB-MET. IHC for the neo-angiogenic marker CD31 demonstrated a low density in the SDHB-MET compared to that observed in a positive control, renal cell carcinoma.[br]In conclusion, the lack of efficacy of sunitinib in the SDHB-MET setting may be attributable to the absence of the tyrosine kinase receptors sunitinib targets, including VEGFR2, on tumor vessels.[br][br]Nothing to Disclose: JM, MM, HT, T-TH, US, SG, EK, MM, ATF, KP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2214 303 2203 MON-296 PO43-02 Monday 1872 2012


1868 ENDO12L_MON-297 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Impact of Ribonucleotide Reductase Large Subunit (RRM1) Gene Expression on Metabolization and Activity of Mitotane in Adrenocortical Cancer Cell Lines Silvia De Francia, Antonina Germano, Ida Rapa, Marco Volante, Federica Laino, Eleonora Duregon, Alfredo Berruti, Mauro Papotti, Massimo Terzolo University of Turin, Orbassano (TO), Italy BACKGROUND[br]Mitotane (o,p[apos]DDD) is the reference therapy for adrenocortical carcinoma (ACC) and should undergo metabolization in o,p[apos]DDE and o,p[apos]DDA to exert its anti-proliferative activity. We have previously shown that there is a link between RRM1 gene expression and o,p[apos]DDD activity in an adjuvant setting. Aim of this study was to assess whether RRM1 gene expression is correlated to the bioavailability and cytotoxic activity of o,p[apos]DDD and its metabolites in ACC cell lines.[br]METHODS[br]The ACC cell lines SW-13 and H295R were incubated with o,p[apos]DDD, o,p[apos]DDE, o,p[apos]DDA at therapeutic concentrations, and cell viability (evaluated using the WST-1 method) was correlated with RRM1 gene expression (evaluated using real time[ndash]PCR). The intracellular concentrations of o,p[apos]DDD and o,p[apos]DDE were evaluated in cells lysates using HPLC-UV method in both cell lines.[br]RESULTS[br]In H295R cells, o,p[apos]DDD, o,p[apos]DDE and o,p[apos]DDA showed a similar cytotoxic effect (IC50 of 32[micro]M, [lt]1[micro]M, 292 [micro]M, respectively) and RRM1 gene expression was not influenced by any drug. In SW-13 cells, o,p[apos]DDD and o,p[apos]DDE were effective at high concentrations only (IC50 of 829[micro]M,122[micro]M, respectively), while o,p[apos]DDA showed a greater cytotoxic activity (IC50 292[micro]M). An up-modulation of RRM1 mRNA was induced by o,p[apos]DDD and o,p[apos]DDE, whose intracellular concentrations were lower in SW-13 than in H295R cells. The higher sensitivity of SW-13 cells to o,p[apos]DDA was associated with the loss of up-modulation of RRM1 mRNA. Moreover, RRM1 gene silencing in SW13 cells increased the intracellular concentrations of o,p[apos]DDD and o,p[apos]DDE, and their cytotoxic activity as well.[br]CONCLUSIONS[br]The present data suggest that RRM1 gene expression may affect both pharmacokinetic and anti-neoplastic activity of o,p[apos]DDD in SW-13 cells.[br][br]Nothing to Disclose: SDF, AG, IR, MV, FL, ED, AB, MP, MT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 497 303 2204 MON-297 PO43-02 Monday 1873 2012


1869 ENDO12L_MON-298 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Expression of Growth Hormone, Prolactin and Their Receptors in the National Cancer Institute[apos]s 60 Cell-Line Panel (NCI60) Elahu Sustarsic Gosney, Riia K Junnila, James Herpy, John J Kopchick Ohio University, Athens, OH; Ohio University, Athens, OH; Ohio University Heritage College of Osteopathic Medicine, Athens, OH In 2012, an estimated 577,190 Americans will die of cancer, making it a leading cause of death in the United States (1). There will also be over 1.6 million new cases of cancer this year. While hundreds of tumor suppressors and oncogenes have been identified, inherited mutations account for a mere 5-10% of all cancers (1). An emerging concept is that a permissive environment must exist that leads to or permits the initiation and progression of carcinogenesis (2). There is a great need to identify the biological processes that establish this permissive environment. The dysregulation of hormones that control cell growth, proliferation and apoptosis may be an important factor and is thought to contribute to carcinogenesis. Many cancer cells express and respond to a wide range of hormones, including growth hormone (GH), which was first detected in human cancer tissue in 1972 (3). In more recent years, accumulating evidence has implicated GH, prolactin (PRL) and insulin/IGF signaling in carcinogenesis. A recent GWAS study of breast cancer found the GH pathway to have the 3rd highest association with breast cancer susceptibility (4). The National Cancer Institute has developed a human cell panel of 60 different tumor-derived cell lines representative of nine types of cancer. Our goal is to identify cancer cell lines in the NCI60 panel that respond to GH, and to treat those cells with GHR and PRL receptor (PRLR) antagonists. We have determined the expression of major components of the GH and PRL signaling pathways in these cell lines including GH, GHR, PRL and PRLR (RNA and protein from all NCI60 cell lines has been provided by NCI). A variety of receptor expression patterns have been detected, including cell lines that express high levels of GHR alone, PRLR alone, both or neither. In some cell lines, these receptors are accompanied by expression of GH or PRL, which opens intriguing questions into the role of autocrine/paracrine activity in these tumor cells. In addition, we have found expression of specific GH and GHR variants in some of the cell lines. Cell lines that express high levels of GHR, PRLR, or both will be treated with GHA and monitored for changes in growth rates. Cell lines that respond will be further analyzed to determine which intracellular signaling pathways are involved. We hope this will allow us to eventually identify specific types of human cancer that might respond to targeted therapeutic interventions that block GHR or PRLR signaling.[br][br](1) American Cancer Society. Cancer Facts [amp] Figures 2012 Atlanta: American Cancer Society, 2012. (2) Holly JM and Perks CM. Cancer as an endocrine problem Best Pract Res Clin Endocrinol Metab 22: 4: 539-550, 2008. (3) Beck C and Burger HG. Evidence for the presence of immunoreactive growth hormone in cancers of the lung and stomach. Cancer 30: 1: 75-79, 1972. (4) Menashe I, Maeder D, Garcia-Closas M, Figueroa JD, Bhattacharjee S, Rotunno M, Kraft P, Hunter DJ, Chanock SJ, Rosenberg PS and Chatterjee N. Pathway Analysis of Breast Cancer Genome-Wide Association Study Highlights Three Pathways and One Canonical Signaling Cascade Cancer Res 2010.[br][br]Sources of Research Support: This work was supported in part by the State of Ohio[apos]s Eminent Scholar Program that includes a gift from Milton and Lawrence Goll, by the AMVETS, by the Diabetes Research Initiative at Ohio University, and by NIH grants DK083729, AG031736.[br][br]Nothing to Disclose: ESG, RKJ, JH, JJK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1956 303 2205 MON-298 PO43-02 Monday 1874 2012


1870 ENDO12L_MON-299 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) The Role of Steroid Receptor, Steroidogenic Enzymes and Prostatic Acid Phosphatase Immunohistochemical Expression in the Diagnosis and Prognosis of Prostate Cancer Dara Hope Cohen, Shen Yao, Yasuhiro Nalamura, Keely McNamara, Hironubu Sasano, Alexander Kirschenbaum, Alice C Levine Mount Sinai School of Medicine, New York, NY; Tohoku University, Sendai, Japan; Mount Sinai School of Medicine, New York, NY [bold]Background: [/bold]Prostate cancer (PCa) is often a multifocal, multiclonal disease with a variable clinical course. Although Gleason score and prostate specific antigen (PSA) are widely used as diagnostic and prognostic markers, they do not reliably predict aggressivity and metastatic potential of PCa. Recent studies have demonstrated steroidogenic enzyme expression in some metastatic PCa clones leading to intratumoral androgen synthesis. In addition, multiple reports demonstrate that prostatic acid phosphatase (PAP) may be a better predictor of bone metastases than PSA.[br][bold]Case/Methods[/bold]: A 70 year-old male presented with a right prostatic nodule noted on exam, a normal PSA and a biopsy demonstrating PCa. The patient underwent a laparoscopic radical prostatectomy and final pathology demonstrated a high grade PCa Gleason 9 (5+4), as well as a smaller, more differentiated lesion, Gleason 7 (4+3). One year later, the patient was found to have an increasing PSA. PET scan demonstrated a lesion in the pleura. Biopsy of the pleural lesion revealed a papillary carcinoma of unknown origin (PSA staining negative) which was subsequently confirmed to be metastatic PCa by positive staining for PAP. Serum PSA levels initially fell after resection of the pleural lesion but then began to rise and the patient was treated with anti-androgen therapy. PET scan detected a solitary 4 cm right pelvic lymph node which was resected and stained negatively for PSA but positive for PAP and AR. Immunohistochemical staining of the primary lesions, the pleural met and lymph node for PSA, PAP, AR, steroidogenic enzymes, ER-alpha, ER-beta, MUC1, VEGF and Glut1 were compared.[br][bold]Results: [/bold]Immunohistochemical features of the pleural and lymph mets were most consistent with the more aggressive primary PCa clone (Gleason 9) including positive staining for PAP, AR, MUC1, ER-beta, Glut1 and VEGF, with minimal PSA immunoreactivity. In addition, both the more aggressive primary clone and the pleural and lymph node mets demonstrated expression of steroidogenic enzymes including Type I 5 alpha reductase and HSD3B1, although expression was more pronounced in metastatic lesions.[br][bold]Conclusion:[/bold] Metastatic PCa may have minimal expression of PSA and additional staining for PAP may be needed to confirm the diagnosis. In addition to PAP, expression of steroidogenic enzymes, ERbeta, MUC1, Glut1 and VEGF in primary PCa clones may help predict biologic and metastatic behavior and guide therapy choices.[br][br]Nothing to Disclose: DHC, SY, YN, KM, HS, AK, ACL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 205 303 2206 MON-299 PO43-02 Monday 1875 2012


1871 ENDO12L_MON-300 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Yin Yan Role of Estrogenic Signaling in Human Malignant Cell Growth: Concomitant Upregulation of Estrogen Receptor-, 12-, 15-Lipoxygenase and 1a-Hydroxylase 25-Hydroxyvitamin D mRNA Expression Dalia Somjen, Fortune Kohen, Sara Katzburg, Orli Sharon, Esther Knoll, Naftali Stern Tel Aviv Sourasky Medical Center and Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel; The Weizmann Institute of Science, Rehovot, Israel Estrogen receptors (ERs) are expressed in various cancerous cells outside the reproductive system. Some cancer types also express 1a- hydroxylase 25- hydroxyvitamin D (1OHase) mRNA such that the 1,25(OH)[sub]2[/sub]D[sub]3 [/sub](1,25) formed can attenuate tumor cell proliferation via autocrine/paracrine effects. Finally, lipoxygenase (LO) enzyme expression has been detected in pancreatic and other cancer cell types, and their products 12 and 15hydroxyeicosatetraenoic acid (HETEs) can enhance tumor cell survival. We investigated whether E[sub]2[/sub], biochainin A (BA), the carboxy- derivative of BA (cBA) and dihydrotestosterone (DHT) affect these systems in adrenal (H295R), colon (320D), ovarian (A2870) and prostate (C4-24) carcinoma cell lines. All cell types expressed ERa, ERb mRNA, 1OHase and 15LO mRNA and produce 15HETEs. There was a general abundance of ERb mRNA expression, such that the ratio of ERb to ERa mRNA was 1.8:1, 30:1, 292:1, 71:1 in H295R, 320D, A2870, and C4-24 cells, respectively. Only DHT and cBA increased ERa in 320D and H295R, while BA decreased it in 320D. DHT increased ERb in A2780, 320D and H295R and inhibited it in C4-24. E[sub]2[/sub] stimulated ERb in A2780 and in C4-24. BA inhibited ERb in A2780 and H295R and stimulated it in 320D and C4-24. cBA inhibited ERb in A2780 and C4-24 and stimulated it in 320D and H295R. 1OHase was inhibited by all hormones in C4-24 whereas in 320D all compounds except BA stimulated it. In A2780 BA inhibited 1OHase and DHT and cBA stimulated in. In 320D all hormones except BA stimulated 1OHase while in H295R E[sub]2[/sub] and BA inhibited it and cBA stimulated. In H295R all hormones inhibited 15HETEs while in A2780 all hormones except DHT stimulated it. In C4-24 all hormones except BA stimulated 15HETEs and in 320D DHT inhibited it while BA and cBA stimulated it. In conclusion several human cancer cell lines express mRNA for ERs, 1OHase and LOs which are modulated by estrogens and DHT. This might form the basis for the use of cancer type-specific estogen/androgen agonists/antagonists and affinity drug targeting, especially via ERb, whose blockade was shown by our group to induce cancer cell apoptosis [italic]in vitro[/italic] (J. Med Chem. 50: 6405- 6410, 2007). These results may suggest that endogenous estrogens may affect thyroid cancer cell growth via opposing pathways: cell growth acceleration via induction of ER and LO expression, in association with the induction of 1OHase thus facilitating the synthesis of 1, 25 which is known to inhibit cell proliferation.[br][br]Nothing to Disclose: DS, FK, SK, OS, EK, NS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 341 303 2207 MON-300 PO43-02 Monday 1876 2012


1872 ENDO12L_MON-301 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Heterozygous Deletion in [italic]Foxd3[/italic] Alters Morphology of Interstitial Cells of Cajal of the Gastrointestinal Tract with Megaesophagus, in Mice Eva Szarek, Matthew F Starost, Mones Abu Asab, Louis Dye, Evan R Ball, Anelia Horvath, Maria Nesterova, Constantine A Stratakis Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, MD; National Institutes of Health (NIH), Bethesda, MD; Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, MD; Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, MD Forkhead transcription factor, [italic]Foxd3[/italic], plays a critical role during embryonic development by controlling lineage specification of neural crest cells and represses melanocyte development. Understanding the molecular mechanisms of [italic]Foxd3[/italic] is crucial for manipulating stem cells for the treatment of disease. Recently, our group identified genetic defects in [italic]FOXD3[/italic] in patients with non-[italic]KIT/PDGFR[/italic] A gastrointestinal stromal tumors (GISTs), or wild-type (WT)-GISTs. GISTs are rare tumors of the gastrointestinal tract (GIT) and arise from interstitial cells of Cajal (ICCs), pacemaker cells of the GIT. WT-GISTs are identified by an absence of c-[italic]KIT[/italic] or [italic]PDGFRA[/italic] gene mutations. For normal development, ICCs and melanocytes require c-[italic]KIT[/italic]. An increase in c-[italic]KIT[/italic] activity can lead to hyperpigmentation and GISTs. [italic]foxd3[/italic] mutant zebrafish reveal a hyperpigmenting phenotype, also observed in patients with GISTs. Given the lack of information on WT-GISTs we utilized a mouse model with altered [italic]Foxd3[/italic] expression. [italic]Foxd3[sup]-/-[/sup] [/italic] mouse embryos fail just after implantation, at approximately 6.5 days (d), because the cells of the inner cell mass cannot be maintained [italic]in vitro[/italic], and blastocyst-derived stem cell lines cannot be established. Therefore, we used [italic]Foxd3[sup]+/- [/sup][/italic] mice that were mated with wild-type breeders. Mice at 6, 12 and 15-months (mo) of age were presented for necropsy, with specific focus on the GIT. Additionally, a section of the ileum was saved for electron microscopy (EM) studies; this was used to examine the morphology of the ICCs. Fewer ICCs were observed in the ileum in EM sections from [italic]Foxd3[sup] +/-[/sup] [/italic] mice. We found no abnormalities at 6- and 12-mo in the GIT, at the histological level. However, at 15-mo, [italic]Foxd3[sup] +/-[/sup][/italic] mice presented with megaesophagus. If [italic]Foxd3[/italic] plays a role in the function of ICCs, then loss of that function could potentially contribute to weakening of the esophageal wall, or poor contractility, with the potential for food to become lodged in the esophageal lumen causing it to dilate. We are currently elucidating the role of [italic]Foxd3[/italic] in ICC development in both mice and zebrafish.[br][br]Nothing to Disclose: ES, MFS, MAA, LD, ERB, AH, MN, CAS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2064 303 2208 MON-301 PO43-02 Monday 1877 2012


1873 ENDO12L_MON-302 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Novel Hematopoietic Neoplasms in Prkar2a-Deficient Mice Emmanouil Saloustros, Sisi Liu, Lina Gugliotti, Anelia Horvath, Kitman Tsang, Maria Nesterova, Herbert C Morse III, Constantine A Stratakis National Institute of Child Health and Human Development, Bethesda, MD; National Institute of Allergy and Infectious Diseases, Bethesda, MD Background: Histiocytic sarcoma (HS) is an aggressive hematological neoplasm that responds poorly to therapy. The molecular etiology and pathology of this disease remain unclear, hampering the development of an effective therapy, and there remains a need for more, and more realistic, animal models. Lymphoproliferative disorders have been reported in mice deficient for the Prkar1a gene coding for the regulatory subunit type 1A of protein kinase A (PKA), but nothing is known about the role of type II PKA regulatory subunits in hematologic malignancies.[br]Methods: Mice deficient for the Prkar1a and Prkar2a alleles have been previously reported (1,2) and were kept on a mixed genetic background (C57BL/129Sv). Mice were crossed to create Prkar2a+/- and Prkar2a-/-. Mice were phenotyped at the age of 3-6-9-12-18 months or when they exhibited signs of advanced disease. Tissues were collected for histological and molecular analysis.[br]Results: Unexpectedly, mice deficient for the Prkar2a allele(s) developed lymphomas, with significant higher incidence compared to the Prkar1a+/- mice [8/13 (61.5%) of the Prkar2a+/- and 5/8 (62.5%) of the Prkar2a-/- vs 3/21(14.2%) of the Prkar1a+/-, Fisher[apos]s exact test p=0.02]. Histologic studies of sections stained with H[amp]E revealed a variety of pathologic changes in lymphoid and non-lymphoid tissues. Lesions characteristic of histiocytic sarcoma (HS) were found in spleen and abdominal lymph nodes, sometimes associated with leukemic infiltrates in lung and bone marrow (BM). Abnormal megakaryopoiesis in spleen and BM was associated with cells actively phagocytosing red cells, others with a pseudo Gaucher-like appearance and increased immature forms. Erythropoiesis was reduced in BM and spleen. Vascular thrombi were associated with schistocytosis. Accumulations of mostly mature plasma cells and some plasmablasts were present in the splenic red pulp together with large, active germinal centers. Cells with characteristics of diffuse large B cell lymphoma often occupied expanded splenic follicles and lymph nodes. Kidneys exhibited degenerative changes in glomeruli and tubules. Median age at presentation was 18 months (range 16-25 months).[br]Conclusions: Our data indicate that R2a may potentially be involved in the development of malignant lesions of the hematologic lineage.[br][br](1) Kirschner LS et al.,Cancer Res. 2005;65:4506. (2)Burton KA et al., Proc Natl Acad USA 1997;94:11067.[br][br]Nothing to Disclose: ES, SL, LG, AH, KT, MN, HCM, CAS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2139 303 2209 MON-302 PO43-02 Monday 1878 2012


1874 ENDO12L_MON-303 POSTER SESSION: Biomarkers, Metastasis [amp] Tumor Progression, [amp] Tumorigenesis - II (1:30 PM-3:30 PM) Potential Role of COX-2 Inhibitor Celecoxib as a Molecular Targeted Therapy in Fibrous Dysplasia and Other cAMP/PKA-Induced Tumors Emmanouil Saloustros, Sisi Liu, Nisan Bhattacharyya, Edward Mertz, Maria Nesterova, Maria De Luz Sierra, Keil Meg, Sergey Leikin, Constantinos Petrovas, Micheal Collins, Constantine A Stratakis National Institute of Child Health and Human Development, Bethesda, MD; National Institute of Dental and Craniofacial Research, Bethesda, MD; National Institute of Child Health and Human Development, Bethesda, MD; National Institute of Allergy and Infectious Diseases, Bethesda, MD Background: Mice double heterozygous for prkar1a and prkaca mutated alleles (prkar1a+/-prkaca+/-) develop multiple tail bone tumors. In these tumors with defective cyclic AMP-dependent protein kinase (PKA) regulation, prostaglandin E2 (PGE2) leads to increase of cAMP, activation of Wnt signaling through the inflamasome, and tumor growth (1). Histological analysis showed similarities between bone tumors from prkar1a+/-prkaca+/- mice and patients with fibrous dysplasia (FD). We sought to study the effect of PGE2 release inhibition, using the selective cyclooxygenase-2 (COX-2) inhibitor celecoxib, in cAMP-induced mice and human bone lesions.[br]Methods: Mice were fed with either standard (n=11) or celecoxib supplemented diet (1,5 gr/kg of NIH-31 diet) (n=11) for 6 weeks. Calcein (10 ug/gr) was administrated subcutaneously 16 and 7 days before sacrifice. Tumors were collected for histological and molecular analysis. Based on calcein fluorescence observed under microscope, the apposition rate of cortical bone at periosteal side was measured. A primary cell culture was established from a tumor of patient with an aggressive form of FD that, like the tumors from the mice, showed abundant cartilage formation. Cells were exposed to different concentrations of celecoxib and cell viability was assessed using the MTT assay. A flow cytometry assay was applied for the simultaneous assessment of the viability (aqua-viability dye) and cycle progression (BrdU, Ki-67 and 7AAD) of the cells in the absence or presence of celecoxib. [br]Results: Celecoxib reduced the rate of tumor growth by two-fold as measured by calcein labeling (p=0.016) and the disorganization of bone material. Treatment reduced PKA enzymatic activity in the tumors as measured by HPLC (4.28 vs 3.37 CMP/ug protein, p=0.07) whereas cAMP levels and total PDE activity did not change. The expression of all inflammasome-related genes (ets-1, caspase-1, il1b and cox2) were down-regulated both at the mRNA and protein level. Similar to the mice, celecoxib demonstrated activity in cells from human FD tumor with IC50 of 50 uM. FACS analysis confirmed the effectiveness of the medication most probably through a cytotoxic mechanism.[br]Conclusion: Celecoxib demonstrated activity in cAMP tumors from both mice and humans. Since local tumor control is essential for patients with FD and other cAMP-induced tumors, celecoxib may merit consideration as the first molecularly targeted therapy in this setting.[br][br]Almeida MQ et al., Hum Mol Genet 2011;20:165.[br][br]Nothing to Disclose: ES, SL, NB, EM, MN, MDLS, KM, SL, CP, MC, CAS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1640 303 2210 MON-303 PO43-02 Monday 1879 2012


1875 ENDO12L_MON-313 POSTER SESSION: Endocrine Nurses[apos] Society (1:30 PM-3:30 PM) Nurse-Led Telephone Consultation Offers Patients with Graves Disease a Satisfying Means of Follow-Up Care Louise A Breen, Dulmini Kariyawasam, Barbara M McGowan, Jake K Powrie, Paul V Carroll Guy[apos]s and St Thomas[apos] NHS Foundation Trust, London, UK Traditionally follow-up care for patients with Graves[apos] Disease (GD) is undertaken via Face to Face consultation in the Secondary Care Setting. First-line treatment in the UK is with anti-thyroid drug therapy which requires patients to attend outpatient review at 4-12 weekly intervals for monitoring of thyroid function tests.[br]The frequency of outpatient reviews can be onerous for patients. The use of the Telephone Consultation is becoming more common place in secondary care and was viewed as a model of care that would provide flexibility and convenience to the patient with GD whilst maintaining patient satisfaction.[br]We evaluated patient experience and satisfaction of the new Telephone Clinic and compared this to traditional Face to Face follow-up. Picker Institute validated questions were used to assess patient experience.[br]Results: Patients agreed they understood the explanations given on the rationale for treatment: Telephone 100% (n=34) versus Face to Face 98% (n=52). Patients agreed they were aware of their management plan: Telephone 97% (n=34) versus Face to Face 89% (n=42). Patients agreed they knew who to contact if they had any concerns: Telephone 97% (n=34) versus Face to Face 68% (n=37). Overall general satisfaction was high with no negative responses recorded: Telephone Satisfaction: Excellent 57% (n=20), Very Good 29% (n=10), Good 11% (n=4). Face to Face Satisfaction: Excellent 46% (n=25), Very Good 30% (n=16), Good 18% (n=10) and Fair 6% (n=3).[br]The results demonstrate Telephone Consultation meets the needs of patients with GD and is not inferior to traditional Face to Face follow-up in terms of patient satisfaction. Further evaluation of the clinical effectiveness of Telephone Consultation is required.[br][br]Nothing to Disclose: LAB, DK, BMM, JKP, PVC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2012 304 2211 MON-313 PO100-01 Monday 1880 2012


1876 ENDO12L_MON-314 POSTER SESSION: Endocrine Nurses[apos] Society (1:30 PM-3:30 PM) Detecting Post-Surgical Recurrence in a Patient with Cushing Disease: A Clinical Nurse[apos]s Perspective Kelley J Moloney, William H Ludlam, Jennifer U Mercado, Marc R Mayberg Swedish Neuroscience Institute, Seattle, WA [bold]Objective:[/bold] To illustrate the complexities of identifying patients with post-surgical refractory Cushing[apos]s disease (CD), we present a patient with conflicting biochemical test results that was later proven to have a recurrence. As clinical decision makers with increasing patient responsibilities in the US healthcare system, the role of the nurse practitioner has become increasingly important in diagnosing recurrent CD.[br][bold]Case presentation: [/bold]A 22-year-old female was initially diagnosed with CD five years earlier. At initial presentation, biochemical tests revealed elevated urinary free cortisol (UFC; 126.5 [mu]g/day, normal [lt]80) and plasma ACTH (61 pg/mL, normal [lt]48), and a positive DEX/corticotropin-releasing hormone test (Dex/CRH; serum cortisol 6.0 [mu]g/dL 15 min after CRH). A 1.1-cm hypoenhancing lesion in the pituitary was identified on MRI. The patient underwent surgical removal of the tumor and tissue was confirmed as corticotroph adenoma. Post-operative serum cortisol levels decreased to 1.1 [mu]g/dL, and the patient reported clinical improvements [ndash] including a 55-lb weight loss, suggesting complete tumor removal and remission of CD. Within four years of surgery, the patient experienced increased weight, acne, fatigue, and proximal muscle weakness. A positive Dex/CRH test (serum cortisol 7.9 [mu]g/dL 15 min after CRH) was reported; however, UFC remained within normal limits (38 [mu]g/day, normal [lt]50). The patient underwent repeat pituitary surgery and corticotroph adenoma was removed. Following surgery, the patient[apos]s serum cortisol decreased to 0.9 [mu]g/dL, consistent with complete removal of the tumor. The patient is currently in remission.[br][bold]Discussion: [/bold]Although UFC testing is commonly used in the diagnosis of CD, recent clinical practice guidelines confirm that normal UFC results should not be cause to dismiss the diagnosis of CD if clinical suspicion is high.[sup]1[/sup] Moreover, diagnosis of CD recurrence should not be based solely on UFC test results. In addition to surgery, treatment choices for recurrent CD may soon include novel medical options (e.g., pasireotide, mifepristone) that may improve patients[apos] symptoms and quality of life.[br][bold]Conclusion:[/bold] This case illustrates that, as clinical decision makers, nurse practitioners should be aware of diagnostic limitations of testing methods, and utilize multiple overlapping biochemical tests to establish CD recurrence. Clinical signs and symptoms should also be heeded [ndash] even with conflicting biochemical test results.[br][br]Nieman et al. J Clin Endocrinol Metab 2008;93:1526-1540.[br][br]Disclosures: WHL: Speaker, Pfizer, Inc., Novartis Pharmaceuticals, Ipsen; Medical Advisory Board Member, Corcept Therapeutics, Novartis Pharmaceuticals, Ipsen, Endo Pharmaceuticals Inc. Nothing to Disclose: KJM, JUM, MRM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1694 304 2212 MON-314 PO100-01 Monday 1881 2012


1877 ENDO12L_MON-315 POSTER SESSION: Endocrine Nurses[apos] Society (1:30 PM-3:30 PM) Monitoring Patient Improvement Parameters Following Medical Treatment of Cushing Disease: A Clinical Case Report Chris Yedinak, Jessica Brzana, Maria Fleseriu Oregon Health [amp] Science University, Northwest Pituitary Center, Portland, OR; Oregon Health [amp] Science University, Northwest Pituitary Center, Portland, OR [bold]Background[/bold]: In the largest Phase III study in patients with Cushing[apos]s disease (CD), pasireotide reduced urinary free cortisol (UFC) in the majority of patients. Improvement in clinical signs and symptoms of CD are important outcomes and particularly relevant to effective nursing care. We report a patient case from this study that illustrates improvements in both biochemical and clinical measures.[br][bold]Case presentation[/bold]: A 24-year-old male patient presented with signs and symptoms of CD in 2004. The patient was obese (149.2 kg; BMI=45.47) and had refractory hypertension (160/85 mmHg) and impaired glucose tolerance treated with glypizide. He had suffered several non-traumatic vertebral compression fractures. Initial screening showed elevated 24-hour UFC (262 [micro]g) and Dex/CRH results (cortisol=24.3 [mu]g/dL at 15 min; ACTH=115 pg/mL). MRI revealed an 8-mm pituitary tumor. He underwent surgery and had a 2-year remission; his HbA1c was reduced (4.8 mg/dL) and antiglycemics were halted. He had recurrent elevated 24-hour UFC (589 [micro]g) and enrolled in the study (randomized to pasireotide 600 [micro]g sc BID). After 3 months, his 24-hour UFC decreased to 320 [micro]g and the dose was increased to 900 [micro]g BID. Biochemical tests normalized toward the end of the 12-month study (UFC=85 [micro]g; morning ACTH=33 ng/L; serum cortisol=19 [micro]g/dL). Additionally, he experienced a 17-kg weight reduction over the last 2 months of the study and reduced blood pressure (141/85 mmHg) despite discontinuation of two antihypertensive medications. Vertebral BMD improved by 6% over baseline; femoral BMD was unchanged. Improvements in memory, irritability, healing, and bruising began after 3 months of treatment, as evaluated by the CushingQoL questionnaire. The patient reported mild adverse events (AEs) related to study drug, including mild pruritis and erythema at injection site (resolved by study week 2), nausea (resolved when he was instructed to eat immediately after injection), and transient fasting hyperglycemia ([le]300 mg/dL). He responded to treatment with metformin 500 mg BID and this treatment was halted after 2 weeks. Pasireotide was continued at the same dose. At study end, his HbA1c was 6.3 mg/dL.[br][bold]Conclusion: [/bold]In addition to biochemical parameters, clinical signs and symptoms are important measures of CD treatment success. This case illustrates successful long-term medical treatment of CD with pasireotide, with clinical improvements and effective control of AEs, including hyperglycemia.[br][br]Nothing to Disclose: CY, JB, MF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1775 304 2213 MON-315 PO100-01 Monday 1882 2012


1878 ENDO12L_MON-316 POSTER SESSION: Endocrine Nurses[apos] Society (1:30 PM-3:30 PM) A 12-Week Music Aerobic Exercise Increases Peripheral Blood Lymphocytes and Regulatory T Cell Differentiation Kuender D Yang, Shio-Ling Lai, Ya-Tien Lo, Lin Wang, Li-Wei Lin, Shu-Hui Yeh Chang Bing Show Chwan Memorial Hospital, Changhua, Taiwan; Central Taiwan University of Science and Technology, Taichung, Taiwan; Chang Gung University, Kaohsiung, Taiwan; HungKuang University, TaiChung, Taiwan Moderate exercise that causes maximal oxygen consumption between 50% and 70% has long been shown to promote cardiopulmonary functions and perceptional health. We have previously shown that Tai-chi Chuan exercise, a moderate exercise, promoted functional mobility and immune functions among diabetic patients. It remains to be determined whether different moderate exercises have different effects on immune functions. This study was conducted to investigate whether a regular music aerobic exercise (MAE) had similar or different effects on blood leukocyte counts and lymphocyte differentiation before and after a 12-week exercise. Forty middle-age women were recruited for this study, thirty eligible for regularly weekly MAE schedule were enrolled into the experimental group and completed pre- and post-test. The other 10 participants who were not eligible to the MAE schedule were enrolled into the parallel control group without exercise. Results showed that the MAE exercise for 12 weeks did not change platelets or red blood cells but selectively increased lymphocytes (p = 0.02). In analyses of lymphocyte subpopulations, we found that the MAE significantly increased CD4, CD8, CD4CD25 and CB8CD56 cells. The MAE also enhanced the Treg cell polarization with enhanced Foxp3 but not T-bet, Gatat-3 or ROR[gamma]T expression. The parallel control group without exercise revealed no change of leukocyte counts or lymphocyte polarization. To our knowledge, this is the first study to show different moderate exercises have a common effect on enhancement of Treg cell polarization, but varied effects on lymphocyte distribution. It may be worthwhile to encourage moderate exercise for middle-age adults as well as diabetic patients to promote immunity.[br][br]Nieman DC (2000) Exercise effects on systemic immunity. Immunol Cell Biol. 78:496[ndash]501. Pagels P, Raustorp A, Archer T, Lidman U, Alricsson M (2011) Influence of moderate, daily physical activity upon body composition and blood lipid profile in Swedish adults. J Phys Act Health Sep 13. Pahkala K, Heinonen OJ, Simell O, Viikari JS, R[ouml]nnemaa T, Niinikoski H, Raitakari OT (2011) Association of physical activity with vascular endothelial function and intima-media thickness. Circulation 124:1956-1963. Pollock ML, Wilmore JH (1990) Exercise in Health and Disease: Evaluation and prescription for prevention and rehabilitation. 2nd ed. WB Saunders, Philadelphia, pp 380-386. Shore S, Shinkai S, Rhind S, Shephard RJ (1999) Immune responses to training: how critical is training volume? J Sports Med Phys Fitness 39:1-11. Simpson RJ, Florida-James GD, Whyte GP, Guy K (2006) The effects of intensive, moderate and downhill treadmill running on human blood lymphocytes expressing the adhesion/activation molecules CD54 (ICAM-1), CD18 (beta2 integrin) and CD53. Eur J Appl Physiol. 97:109-21. Suzuki K, Nakaji S, Yamada M, Totsuka M, Sato K, Sugawara K (2002) Systemic inflammatory response to exhaustive exercise: cytokine kinetics. Exerc Immunol Rev 8:6-48. Trovati M, Giovanni A (1992) Moderate exercise increase platelet function in type 1 diabetes without severe angiopathy and in good control. Diabetes Care 15:1742-1746. Wang C, Collet JP, Lau J (2004) The effect of TCC exercise on health outcomes in patients with chronic conditions: a systematic review. Arch Intern Med 164:493-501. Weltman A, Weltman J, Rutt R, Seip R, Levine S, Snead D, Kaiser D, Rogol A (1989) Percentages of maximal heart rate, heart rate reserve, and VO2 peak for determining endurance training intensity in sedentary women. Int J Sports Med 10:212-216. Woods JA, Ceddia MA, Wolters BW, Evans JK, Lu Q, McAuley E (1999) Effects of 6 months of moderate aerobic exercise training on immune function in the elderly. Mech Ageing Dev 109:1-19. Yeh SH, Chuang H, Lin LW, Hsiao CY, Eng HL (2006) Regular tai chi chuan exercise enhances functional mobility and CD4CD25 regulatory T cells. Br J Sports Med 40:239-243. Yeh SH, Chuang H, Lin LW, Hsiao CY, Wang PW, Yang KD (2007) Tai Chi Chuan exercise decreases HbA1c level along with increase of regulatory T cells and decrease of cytotoxic T cell population in Type 2 diabetic patients. Diabetes Care 30:716-8. Yeh SH, Chuang H, Lin LW, Hsiao CY, Wang PW, Liu RT, Yang KD (2009) Regular Tai Chi Chuan exercise improves T cell helper function of patients with type 2 diabetes mellitus with an increase in T-bet transcription factor and IL-12 production. Br J Sports Med 43:845-50. Yu HR, Chen RF, Hong KC, Bong CN, Lee WI, Kuo HC, Yang KD (2005) IL-12-independent Th1 polarization in human mononuclear cells infected with varicella-zoster virus. Eur J Immunol 35:3664-3672.[br][br]Sources of Research Support: This study was supported by a grant NSC 100-2314-B-166-004 from the National Science Council, Taiwan; and a grant RA 11027 from Chang Bing Show Chwan Memorial Hospital, Taiwan. Appreciation is extended to all the staff and participants, whose devotion and cooperation made this study possible.[br][br]Nothing to Disclose: KDY, S-LL, Y-TL, LW, L-WL, S-HY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1630 304 2214 MON-316 PO100-01 Monday 1883 2012


1879 ENDO12L_MON-317 POSTER SESSION: Endocrine Nurses[apos] Society (1:30 PM-3:30 PM) Self-Perception of Cognitive Function and Associated Quality of Life (QoL) among Patients with Active Acromegaly (AA), Controlled Acromegaly (CA), and Non-Functional Pituitary Adenoma (NFPA) Chris Yedinak, Maria Fleseriu Oregon Health [amp] Science University, Northwest Pituitary Center, Portland, OR; Oregon Health [amp] Science University, Northwest Pituitary Center, Portland, OR [bold]Background:[/bold] Pituitary adenoma (PA) represents 15% of brain tumors. Cognitive dysfunction has been documented in patients with hormonally active PA, although etiology remains unclear. Herein, we characterize and compare cognitive function and QoL among patients with AA, CA, and NFPA.[br][bold]Methods: [/bold]Using a modified, disease-specific FACT-Cog questionnaire, patients rated their own perception of ability in learning, concentration and distractibility, mental agility, memory and recall, and verbal recall on a scale from 1 (no dysfunction) to 6 (very dysfunctional). All AA and CA patients were post-TSH resection; all NFPA patients were pre-treatment. Patients with AA or CA (control defined as normal IGF-1 for age after surgery and/or serotonin receptor ligand [SRL] therapy with nadir GH[lt]1ng/ml after glucose suppression) had biochemical and/or pathologic confirmation of a growth-hormone[ndash]secreting PA.[br][bold]Findings: [/bold]We surveyed 27 patients with acromegaly (10 AA [4 on SRL]; 17 CA [7 on SRL]) and 14 patients with NFPA. The mean FACT-Cog score for each group showed an overall mild-moderate level of perceived cognitive dysfunction (2.8/6). Prevalence of cognitive dysfunction was 54%, 57%, and 69% in the CA, AA, and NFPA groups, respectively. All 14 NFPA patients reported slowed mental agility ([italic]P[/italic]=0.001) vs 6/17 (35%) CA patients and 5/10 (50%) AA patients ([italic]P[/italic]=0.017). Among AA and CA patients, there was no significant correlation between overall cognitive dysfunction, age at diagnosis, estimated duration of symptoms, cure, SRL use, sex, or hypogonadism. There was no significant difference between patients with NFPA, AA, or CA with respect to perceived ability to learn, mental agility, or verbal recall. However, CA patients had higher QoL scores than either AA patients ([italic]P[/italic]=0.014) or NFPA patients ([italic]P[/italic]=0.022).[br][bold]Conclusion: [/bold]All PA patient groups demonstrated mild-moderate perceived cognitive dysfunction. While there was no significant difference in overall perceived cognitive dysfunction between patients with CA, AA, and NFPA, QoL was rated significantly higher among patients with CA vs those with either AA or NFPA. Further study is needed to compare these results with a culture-, age-, and sex-matched cohort without PA. A longitudinal study may also clarify cognitive changes and treatment benefits among patients with PA.[br][br]Nothing to Disclose: CY, MF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1780 304 2215 MON-317 PO100-01 Monday 1884 2012


1880 ENDO12L_MON-325 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Hematopoietic Progenitors (HP) and Erythropoietin (EPO) Regulate Mesenchymal Commitment and Lead to Decreased Osteogenesis in a Thalassemia Mouse Model Zhiwei Yang, Maybel Yau, Olga Myszko, Adele L Boskey, Stefano Rivella, F Patrick Ross, Maria G Vogiatzi Weill Cornell Medical College, New York, NY; Hospital for Special Surgery, New York, NY Background: Thalassemias are congenital anemias associated with high rates of osteoporosis as a result of decreased bone formation. These diseases are accompanied by increased expansion of defective erythroids and high serum EPO. Since the genetic defect lies in hematopoiesis we hypothesized that the ineffective erythropoiesis in thalassemia affects bone mesenchymal differentiation and osteogenesis.[br]Qbjective: To determine mechanisms by which ineffective erythropoiesis leads to bone loss using the th3/+ mouse model of [beta] thalassemia intermedia, which has the hematological and bone abnormalities seen in patients.[br]Methods: Study groups wt and th3/+ (n=6/group). Mesenchymal stem cells (MSC) were cultured with osteogenic and adipogenic media. MSC cocultures with bone marrow (BM) hematopoietic progenitors (HP) from wt and th3/+ (two chamber system). MSC cultures with a recombinant EPO antibody or an IgG1 control. Osteogenesis was assessed by determining CFU-Ob and Runx2, bone sialoprotein, osterix, osteocalcin expression by qPCR.[br]Results: i) Compared to wt, MSC cultures from th3/+ showed decreased CFU-Ob and ostegenic gene expression). Ii) Osteogenesis was decreased when MSCs were exposed to early HPs (CD4- CD8- B220- CD11b- Gr-1- CD49b- BM cells) from th3/+ mice compared to wt regardless of MSC origin (th3/+ vs. wt). iii) Lin- HP from th3/+ showed similar effect on MSC differentiation. iv) MSC cultures treated with EPO showed a reversal of decreased osteogenesis in th3/+ but not wt, suggesting that EPO acts in a paracrine/autocrine fashion to mediate MSCs changes in thalassemia.[br]Conclusions: Bone loss in th3/+ involves changes in MSC commitment towards decreased osteogenesis. Thalassemia HPs regulate MSC fate. While further studies are under way to determine the specific HPs that are responsible for this effect, our data indicate that Lin- multipotent progenitors are involved. In addition, our data suggest that EPO affects MSC differentiation in th3/+ in a paracrine/autocrine fashion. These findings are being confirmed with additional in vitro experiments. Overall, our results indicate that erythropoiesis in thalassemia decreases osteogenesis through mechanisms involving mesenchymal interactions between HPs and EPO.[br][br]Sources of Research Support: NHLBI.[br][br]Nothing to Disclose: ZY, MY, OM, ALB, SR, FPR, MGV 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1320 305 2216 MON-325 PO04-01 Monday 1885 2012


1881 ENDO12L_MON-326 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) A Mouse Model of Double Heterozygosity for Protein Kinase A Regulatory Subunits Promotes Differentiation of cAMP-Induced Bone Tumors Emmanouil Saloustros, Kitman Tsang, Matthew Starost, Edward Mertz, Sisi Liu, Elena Makareeva, Nadella Kiran, Maria Nesterova, Sergey Leikin, Constantine A Stratakis National Institute of Child Health and Human Development, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institute of Child Health and Human Development, Bethesda, MD Background: The PRKAR1A gene coding for the regulatory subunit type 1A of protein kinase A (PKA) is mutated in Carney complex (CNC) patients, but how tumors form is not well understood. Mice heterozygous for a null allele of prkar1a develop a variety of tumors overlapping those of CNC patients. Deletion of a catalytic subunit allele prkaca+/- on the prkar1a+/- background unexpectedly increased the number and aggressiveness of bone tumors without development of schwannomas or thyroid lesions. In this study we tested the role of type II PKA regulatory subunits in tumors of the bone and other tissues.[br]Methods: Double heterozygous mice (prkar1a+/-prkar2a+/- and prkar1a+/-prkar2b+/-) were generated by crossing the single heterozygous mice for the prkar1a, prkar2a and prkar2b mutated alleles. The mice were phenotyped at the age of 3, 6, 9, 12 and 18 months and compared to prkar1a+/- mice. Tumors were collected for histology, microscopy and molecular analysis.[br]Results: Both pairs of double heterozygous mice developed bone tumors; later onset of tumors was seen in the prkar1a+/-prkar2b+/- mice. Interestingly, mice knockout or heterozygous for the type II PKA regulatory subunits did not develop tumors neither in bone nor in other tissues. Histology showed abnormal proliferation of a fibroblastoid-like cell with more apoptosis in the bone lesions from double heterozygous mice. Polarized microscopy and Raman micro-spectroscopy revealed that bone material growing in tumors was immature and poorly organized in prkar1a+/- mice (disorganized collagen and osteocytes and undermineralized matrix). Prkar2a+/- deletion on the prkar1a+/- background rescued local organization and mineralization but worsened global organization of cortical tumor bone. The prkar1a+/-/prkar2b+/- deletion rescued both global and local organization and mineralization, resulting in formation of mature bone. Tumors from double heterozygous mice showed higher PKA II activity, mediated by prkacb over-expression. Gene expression analysis and differentiation experiments are ongoing.[br]Conclusion: Prkar1a is major tumor-suppressor gene in the bone. However, prkar2b or prkar2a deficiencies in addition to prkar1a lead to better differentiation of bone tumors, indicating a compensating role for the type II regulatory subunits in cAMP/PKA-induced bone tumors related to PRKAR1A deficiency.[br][br]Nothing to Disclose: ES, KT, MS, EM, SL, EM, NK, MN, SL, CAS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1601 305 2217 MON-326 PO04-01 Monday 1886 2012


1882 ENDO12L_MON-327 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Osteoblasts Derived from MKP-1 Null Mice Exhibit Age-Specific Sexual Dimorphism in the Effects of PTH on IGF-1 mRNA Expression Tanuka Datta, Chandrika D Mahalingam, Abdul B Abou-Samra, Nabanita S Datta Wayne State University School of Medicine, Detroit, MI Parathyroid hormone (PTH) and insulin like growth factor-1 (IGF-1) are important skeletal regulators. PTH increases the production of IGF-1 in osteoblasts in vitro and in vivo; and the anabolic action of PTH requires paracrine and autocrine effects of IGF-1. We have previously demonstrated that PTH up-regulates MAPK phosphatase-1 (MKP-1), an enzyme responsible for dephosphorylation and inactivation of MAPKs, in the process of bone formation. Age-related sexual dimorphism, with osteopetrosis in males and osteopenia in females was shown in MKP-1 knockout (KO) mice. In growing MKP-1 KO animals PTH anabolic effect was attenuated. Disparate PTH effects on in vitro mineralization were observed in primary osteoblasts derived from MKP-1 KO male and female mice. Since both PTH and IGF-1 regulates ERK-MAPK, osteoblast cell proliferation, differentiation and bone formation, we examined if MKP-1is involved in PTH regulation of IGF-1 and bone homeostasis. In this study we evaluated the expression of IGF-1 mRNA in primary calvarial osteoblasts isolated from growing (2-3 week) and older (14-16 week) wild type (WT) and MKP-1 null male and female mice. Calvarial osteoblasts were isolated via sequential collagenase digestion and differentiated with ascorbic acid and b-glycerophosphate with or without PTH. IGF-1 expression was analyzed by real-time PCR. In osteoblasts derived from growing (2-3 week) mice PTH stimulated IGF-1 expression 3- fold (p[lt]0.02) in WT and 5-fold (p[lt]0.002) in KO males, compared to respective untreated controls. In growing females PTH increased IGF-1 expression 4-fold (p[lt]0.04) and 2-fold (p[lt]0.04) in WT and KO osteoblasts respectively compared to untreated cells. Using osteoblasts derived from older (14-16 week) animals PTH treatment showed 6.6-fold (p[lt]0.002) increase in IGF-1 expression in male KO mice. In contrast, IGF-1 expression was down-regulated 2-fold following PTH treatment (p[lt]0.01; 50% of untreated cells) in osteoblasts from KO females. In this age group osteoblasts derived from either male or female WT mice showed no significant differences in IGF-1 expression with and without PTH treatment. These data provide evidence that the effects of PTH on IGF-1 are regulated by MKP-1 and that IGF-1 may be involved in age-specific sexually dimorphic bone phenotype of the MKP-1 KO mice. We propose a cross-talk between MKP-1 and IGF-1 in PTH signaling and bone anabolic action.[br][br]Sources of Research Support: Funding: NIDDK to NSDatta.[br][br]Nothing to Disclose: TD, CDM, ABA-S, NSD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 383 305 2218 MON-327 PO04-01 Monday 1887 2012


1883 ENDO12L_MON-328 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Sex-Specific Rapid Membrane Responses of Rat Costochondral Chondrocytes to 17[beta]-Estradiol Are Estrogen Receptor [alpha]- and [beta]-Dependent Khairat Bahgat El Baradie, Yun Wang, Barbara Boyan, Zvi Schwartz Georgia Institute of Technology, Atlanta, GA; Georgia Institute of Technology, Atlanta, GA Specific receptors for estrogen have been found in both male and female articular cartilage, growth plate and fracture callus. Our work using rat costochondral growth-plate chondrocytes as a model shows rapid increases in PKC activity only in cells from female rats in response to 17[beta]-estradiol (E2). PKC activation is not affected by the nuclear estrogen receptor (ER) antagonist ICI 182780, but can be blocked by antibodies to ER[alpha] and antibodies to ER[beta]. This suggests that non-traditional membrane-dependent mechanisms are involved. The aim of the present study was to investigate which ERs are involved in sex-specific rapid membrane responses to E2 and to investigate if male and female chondrocytes have the same signaling pathway downstream of the receptor. To establish the receptors involved; PKC activity was measured in confluent chondrocytes isolated from both male and female rats that were treated with either DPN (specific activator of ER[beta]), PPT (specific activator of ER[alpha]) or THC (specific activator of ER[alpha] and inhibitor of ER[beta]). To establish the pathway involved in E2-dependent PKC activation, PKC activity was measured in confluent chondrocytes isolated from female rats that were treated with E2 [plusmn] GDP[beta]S (G-protein inhibitor), U73122 (inhibitor of PLC) and AACOCF3 (inhibitor of cytosolic phospholipase A2 [cPLA2]). To determine if the downstream signaling pathway was functional in male cells, male and female cells were treated with PLA2 activating peptide (PLAA) and the PLC activator m-3M3FBS and PKC activity was measured. DPN and PPT induced PKC activity in female cells but not in male cells, while THC had no effect in either type of cells. PKC activation in female cells depended on G protein, PLC and PLA2, based on inhibition by GDP[beta]S, U73122 and AACOCF3. Although activation of PKC by 17[beta]-estradiol was observed only in female chondrocytes, downstream activators PLAA peptide and m-3M3FBS induced a dose-dependent increase in PKC activity in both male and female cells. These results suggest that the rapid membrane response of rat chondrocytes to 17[beta]-estradiol is both ER[alpha] and ER[beta] dependent in female chondrocytes. Although male chondrocytes did not exhibit a rapid response, the downstream components of the signaling pathway were present and active. The different expression levels of ERs or the ability of these receptors to form a complex, might contribute to the sex-specific rapid membrane response between male and female chondrocytes.[br][br]Nothing to Disclose: KBEB, YW, BB, ZS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2172 305 2219 MON-328 PO04-01 Monday 1888 2012


1884 ENDO12L_MON-329 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Characterization of a Truncation Mutation in IRS1 Protein and Its Effect on Skeletal Development Anyonya R Guntur, Victoria E DeMambro, Clifford J Rosen Maine Medical Center Research Institute, Scarborough, ME IRS1 is a critical adaptor protein that is involved in controlling signaling pathways downstream of insulin and IGF-1. We have recently reported on a spontaneous one-nucleotide deletion in the [italic]Irs1[/italic] gene in an inbred C3H mouse strain that results in a truncated protein. C3-[italic]Irs1[sup]sml[/sup]/Irs1[sup]sml [/sup][/italic]are small, lean mice that exhibit reductions in aBMD, femoral cortices, trabecular BV/TV, and serum IGF-1 compared to controls[sup]([bold]1[/bold])[/sup]. Since strain background has been shown to be a modifier of bone phenotypes, we then backcrossed the [italic]Irs1[sup]sml[/sup][/italic] mutation onto a B6 background (n10). B6-[italic]Irs1[sup]sml[/sup]/Irs1[sup]sml[/sup][/italic](B6[italic][sup]sml/sml[/sup][/italic]) male and female mice were than phenotyped by DEXA, [micro]CT and immunohistological techniques and compared to their control littermates. Much like their C3 counterparts, B6[italic][sup]sml/sml[/sup][/italic] mice are small, lean mice with reduced aBMD and a 20% reduction in IGF-1 serum levels. Surprisingly, [micro]CT analysis of the femur revealed that while these mice have reduced cortices, they have no reduction in trabecular bone volume as seen in the C3H strain. Histological and morphological observations showed that the long bones are shorter in the B6[italic][sup]sml/sml[/sup][/italic] compared to the +/+ controls. We observed that the growth plate of the B6[italic][sup]sml/sml[/sup][/italic] was shorter when compared to the +/+ control (postnatal day1). We then performed cell proliferation studies at 18.5dpc to identify potential downstream signaling pathways, preliminary observations indicate that the B6[italic][sup]sml/sml[/sup][/italic] mice have more proliferating cells at this stage in the growth plate. Utilizing antibodies against Col II and Col X proteins, we observed that collagen protein expression in the growth plate was normal in B6[italic][sup]sml/sml[/sup]. [/italic]Using an antibody specific to the C-terminus we probed the chondrocytes and osteoblasts in the long bones and skull base for IRS1 protein. We observed no binding in the B6[italic][sup]sml/sml[/sup][/italic] mice suggesting that they do not express the full length protein. To identify if the truncated protein can be expressed in vitro, we cloned the mutant form of IRS1 and expressed it transiently in MC3T3E1 preosteoblasts and HEK cells. We observed that this truncated form of the protein is present and is responsive to insulin/IGF-1 treatment. Further studies will need to be performed to see if this is due to the fact that the mutant protein retains an intact pleckstrin homology domain. This mouse model will be important for studying the effects of attenuated IR and IGF-1 signaling on bone and also the effects of loss of signaling on whole body metabolism.[br][br](1)DeMambro et al.,JOE 2010.,241-252.[br][br]Nothing to Disclose: ARG, VED, CJR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 723 305 2220 MON-329 PO04-01 Monday 1889 2012


1885 ENDO12L_MON-330 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) The Synergistic Effects of 1[alpha],25(OH)[sub]2[/sub]D[sub]3[/sub] and BMP-2 on Osteoblast Mineralization Are Mediated through Both VDR and Pdia3 Jiaxuan Chen, Jung H Park, Christopher R Dosier, Subhendu De, Asia A Bailey, Robert E Guldberg, Barbara D Boyan, Zvi Schwartz Georgia Institute of Technology, Atlanta, GA 1[alpha],25-Dihydroxyvitamin D3 (1[alpha],25(OH)[sub]2[/sub]D[sub]3[/sub]) and bone morphogenetic protein-2 (BMP-2) have been commonly used to stimulate osteoblastic differentiation of many cell types. 1[alpha],25(OH)[sub]2[/sub]D[sub]3[/sub] regulates osteoblasts through both the classical vitamin D receptor (VDR) mediated genomic pathway and through protein disulfide isomerase family A, member 3 (Pdia3) mediated rapid responses. However, the role of VDR and Pdia3 in osteoblast differentiation by 1[alpha],25(OH)[sub]2[/sub]D[sub]3 [/sub]and interaction between 1[alpha],25(OH)[sub]2[/sub]D[sub]3 [/sub]and BMP-2 are not well known. In this study, we treated wild type, Pdia3-silenced and VDR-silenced pre-osteoblastic MC3T3-E1 cells with either BMP-2, 1[alpha],25(OH)[sub]2[/sub]D[sub]3, [/sub]or both and measured osteoblast marker expression in 2D culture and mineralization in a 3D poly [epsilon]-caprolactone scaffold model. Quantitative PCR was used to measure gene expression, while micro-CT, scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) were used to analyze the mineralized volume, the morphology and the chemical composition, respectively. We found that 1[alpha],25(OH)[sub]2[/sub]D[sub]3[/sub] and BMP-2 caused a synergistic increase in osteoblast marker expression, while silencing of both receptors attenuated this effect. A similar synergistic increase was found with mineralized volume in the 3D model. Silencing of Pdia3 resulted in no response to both factors, while silencing of VDR caused a significant decrease in mineralized volume after 1[alpha], 25(OH)[sub]2[/sub]D[sub]3[/sub] treatment. SEM also showed a unique morphology of mineralized matrix in the group treated with both factors. XPS data showed that combined treatment increased calcium and phosphate content and distribution in wild type cells. These data indicate a synergistic cross talk between 1[alpha],25(OH)[sub]2[/sub]D[sub]3[/sub] and BMP-2 toward osteogenesis, as both VDR and Pdia3 regulated mineralization and also mediated the effect of 1[alpha], 25(OH)[sub]2[/sub]D[sub]3[/sub] and BMP-2.[br][br]Sources of Research Support: Price Gilbert, Jr. Foundation.[br][br]Nothing to Disclose: JC, JHP, CRD, SD, AAB, REG, BDB, ZS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1917 305 2221 MON-330 PO04-01 Monday 1890 2012


1886 ENDO12L_MON-331 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) PLAA Is a Cell-Surface Protein in Osteoblasts and Is Required for Mediating 1[alpha],25(OH)2D3-Induced Rapid PKC Activation Maryam Doroudi, Barbara D Boyan, Zvi Schwartz Georgia Institute of Technology, Atlanta, GA; Georgia Institute of Technology, Atlanta, GA 1[alpha],25-dihydroxy vitamin D3 (1,25D3) regulates osteoblasts via two different mechanisms: vitamin D receptor (VDR)-dependent gene transcription and rapid membrane-signaling via VDR as well as protein disulfide isomerase, family A, member 3 (Pdia3). In membrane-mediated signaling, ligand binding to Pdia3 inside caveolae causes a rapid increase in phospholipase A2 (PLA2) activity leading to release of arachidonic acid and PLC-dependent PKC[alpha] activation. Previous studies using a 21 amino acid PLAA peptide as the activator of PLA2 demonstrated that this peptide mimics the effects of 1,25D3 on osteoblasts. PLA2 inhibitors blocked the effect of PLAA peptide on PLA2, suggesting that the full-length protein acts upstream of PLA2, supporting the hypothesis that PLAA links the Pdia3 receptor complex with PLA2. This study examined the role of PLAA in mediating 1,25D3-dependent PKC activation in osteoblasts. Wild type and PLAA-silenced (shPLAA) MC3T3-E1 osteoblasts were treated with either vehicle or 1,25D3; activities of PLA2 and PKC as well as release of PGE2 were measured. Wild type MC3T3-E1 osteoblasts were treated with either vehicle or 1,25D3 and chemically cross-linked with a non-cleavable, membrane-impermeable crosslinker BS3. Whole cell lysates were analyzed by Western blot. Subconfluent cultures of MC3T3-E1 cells immunostained against PLAA, caveolin-1 (Cav-1) and Pdia3 were imaged using confocal microscopy. Whole cell lysates were collected for immunoprecipitation and caveolae isolation and analyzed by Western blot. PLAA, Pdia3, and Cav-1 were detected in plasma membranes and caveolae. Pdia3-immunoprecipitated samples were positive for PLAA only after 1,25D3 treatment. Cav-1 was detected when immunoprecipitated with anti-Pdia3 and anti-PLAA in both vehicle and 1,25D3 treated cells. These observations were confirmed by immunohistochemistry. 1,25D3 failed to activate PLA2 and PKC or cause PGE2 release in shPLAA cells. PLAA-antibody successfully blocked the PLAA protein and consequently suppressed PKC activity in MC3T3-E1 cells. Crosslinking studies confirmed the localization of PLAA on the extracellular face on the plasma membrane in untreated MC3T3-E1 cells. Taken together, our results suggest that PLAA is an important mediator of 1,25D3 rapid membrane mediated signaling. 1,25D3 initiates changes bringing Pdia3 into proximity with PLAA, and aiding in transducing the signal from caveolae to the plasma membrane.[br][br]Sources of Research Support: Price Gilbert, Jr. Foundation and children[apos]s Healthcare of Atlanta.[br][br]Nothing to Disclose: MD, BDB, ZS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1942 305 2222 MON-331 PO04-01 Monday 1891 2012


1887 ENDO12L_MON-332 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) The Calcemic and Calciuric Response to Oral Calcium Loading Does Not Differ by Reproductive Stage in Women Adapted to a Low Calcium Intake Inez Schoenmakers, Landing MA Jarjou, Gail R Goldberg, Duangporn Harnpanich, Kit Tsoi, Ann Prentice MRC Human Nutrition Research, Cambridge, UK; MRC Laboratories, Keneba, Gambia [bold]Introduction[/bold]: In pregnancy and early lactation parathyroid hormone (PTH) concentrations may be suppressed and uncertainty exists about how calcium (Ca) metabolism is regulated, particularly when Ca-intake is low. In this pilot study, we investigated the calcaemic and calciuric response after an oral Ca-load in pregnant (P), lactating (L) and non-pregnant and non-lactating (NPNL) Gambian women with a low habitual Ca-intake.[br][bold]Methods[/bold]: 30 fasting women (n=10/group) received 1g elemental oral Ca (CaCO[sub]3[/sub]). Blood, plasma (p) and urine (u), was collected pre-dose and at 2, 4 hours (h) and 3h, respectively and analysed (blood: ionised Ca (iCa); p: total Ca (tCa), phosphate (P), creatinine (Cr), PTH, 1,25(OH)[sub]2[/sub]vitamin D (1,25(OH)[sub]2[/sub]D), osteocalcin (OC), [szlig] C-terminal cross-linked telopeptide of type 1 collagen ([szlig]CTX), cyclic adenosine monophosphate (cAMP); u: Ca, P, Cr, cAMP). The fractional excretion of Ca (Ca[sub]e[/sub]), P (P[sub]e[/sub]) and nephrogenic cAMP (NcAMP) were calculated. Within and between group differences were analysed by ANOVA with Scheff[eacute] post-hoc tests.[br][bold]Results: [/bold]Baseline concentrations pPTH and pOC were significantly lower and NcAMP and p1,25(OH)[sub]2[/sub]D higher in P women. In L women, pPTH and p[szlig]CTX were elevated. Groups did not differ in iCa, pP, Ca[sub]e[/sub], P[sub]e[/sub], uCa/Cr and uP/Cr.[br]Post-loading, iCa and ptCa concentrations increased and pPTH, NcAMP and [szlig]CTX decreased in all groups. In all groups uCa/Cr significantly increased and uP/Cr and P[sub]e[/sub] decreased from 2 to 4h post-loading. Ca[sub]e[/sub] only increased in P women. The magnitude of change did not differ between groups for any analyte.[br][bold]Conclusion:[/bold] In pregnant women with a habitually low calcium intake pPTH is suppressed, NcAMP and p1,25(OH)[sub]2[/sub]D are elevated and bone formation is inhibited. Pregnancy induced absorptive hypercalciuria is not observed. In lactating women, pPTH and bone resorption were elevated. Pregnant, lactating and NPNL women responded to a similar extent to Ca-loading. This indicates that pregnant, lactating and NPNL women adapted to a low Ca intake have similar rates of intestinal Ca absorption and extent of renal Ca conservation. The physiological changes in Ca and bone metabolism occurring in pregnancy and lactation appear not to lead to a further increase in Ca absorption and renal Ca reabsorption as observed in Western women with Ca intakes close to recommendations.[br][br]Sources of Research Support: Medical Research Council.[br][br]Nothing to Disclose: IS, LMAJ, GRG, DH, KT, AP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 919 305 2223 MON-332 PO04-01 Monday 1892 2012


1888 ENDO12L_MON-333 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Vitamin D Receptor Promoter Genotype Is Predictive of Changes in Fibroblast Growth Factor 23 with Treatment of X-Linked Hypophosphatemic Rickets Mary D Ruppe University of Texas Medical School at Houston, Houston, TX X-linked hypophosphatemic rickets (XLH) is the most common genetic form of hypophosphatemic rickets with an incidence of 3.9-5 per 100,000 live births. Resulting in lower extremity deformities, bone or joint pain, short stature and dental abnormalities, XLH is characterized by abnormal regulation of phosphate homeostasis and vitamin D metabolism with fibroblast growth factor 23 (FGF23) being the leading contributor to the abnormal phosphate homeostasis. Recent work has shown that patients with XLH have an increase in FGF23 once on therapy. The factors that contribute to this increase in FGF23 are poorly understood. The objective of this study was to create a prediction model for treatment associated changes in FGF23. At outpatient clinic visits, a history was taken, a clinical examination was performed and laboratory evaluation was obtained. Data on mutational status, inheritance pattern, kindred membership, sex, growth velocity, medication doses along with measurements of serum calcium, creatinine, phosphorus, alkaline phosphatase, intact PTH, FGF23 and urinary TMP/GFR were obtained. FGF23 levels were measured using an intact assay (Kainos, Japan). DNA was isolated. The haplotype structure of the human VDR promoter region was determined by genotyping rs4516035 (rs45) and rs11568820 (rs11) polymorphisms. A mixed effects model was developed to explore the relationships between the change in FGF23 with treatment and familial, clinical and laboratory parameters. The model utilized clustering at the level of the subject. Data from 22 subjects who had FGF23 levels obtained both on and off treatment were included in the analysis. During the study period, each subject visited the clinic between 2 and 13 times. The data was analyzed using STATA version 10.0 (StataCorp LP, College Station, TX). Utilizing the developed statistical model, there was significant relationship between the genotype of rs11 (p=0.009) and not that of rs45 (p= 0.319) and the fold change in FGF23. Additionally, there was an association between the serum phosphorus (p= [lt] 0.0001) and iPTH (p= 0.02) with the change in FGF23. There appeared to be no relationship between phosphorus dose, calcitriol dose, other laboratory parameters, growth parameters or inheritance factors. This data highlights the complex factors that underlie the regulation of FGF23 in a clinical setting and adds to our understanding of this process.[br][br]Nothing to Disclose: MDR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2327 305 2224 MON-333 PO04-01 Monday 1893 2012


1889 ENDO12L_MON-334 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Growth Plate Chondrocytes Dedifferentiate in Monolayer but Redifferentiate in High-Density Pellet Culture, Sequentially Expressing Resting, Proliferative, and Hypertrophic Zone Markers Stephan Stanislaw Spath, Anenisia C Andrade, Michael Chau, Ola Nilsson Karolinska Institutet and University Hospital, Stockholm, Sweden Longitudinal bone growth occurs at the growth plate, which is located near the ends of long bones. At the growth plate, cartilage expansion, through chondrocyte proliferation, hypertrophy and matrix production results in elongation of the bone. This process is orchestrated by the interaction of a large number of endocrine and paracrine factors. The study of these complex interactions has been limited by the lack of well characterized chondrocyte culture models and that growth plate chondrocytes cultured in mono-layer rapidly de-differentiate into fibroblast-like cells with high expression of Collagen type I (Col1) and minimal expression of chondrocytic markers, Collagen type II (Col2) and X (Col10).[br]Dedifferentiated rat growth plate chondrocytes in monolayer were trypsinized and centrifuged to form high-density pellets. The cells rapidly (within 3 days) redifferentiated into Col2-expressing chondrocytes as evidenced by a more than 200-fold increase in Col2 expression, up to a level of expression similar to that of growth plate chondrocytes[italic] in vivo[/italic]. We then assessed expression of newly identified resting (Sfrp5) and proliferative zone (Prelp) markers. With prolonged culture, these chondrocytes underwent a reproducible differentiation program characterized by an early rise in expression of resting zone marker Sfrp5, followed by a rise in the proliferation marker Prelp, and finally an increase in hypertrophic zone markers Indian hedgehog and Col10 expression.[br]Our findings suggest that growth plate chondrocytes cultured in high-density pellets undergo a sequential differentiation program similar to growth plate chondrocytes [italic]in vivo[/italic]. These findings may thus help to improve [italic]in vitro[/italic] studies aimed at clarifying the interactions of endocrine and paracrine factors in the regulation of longitudinal bone growth at the growth plate.[br][br]Sources of Research Support: This work was supported by grants from the Swedish Research Council (K2007-52X-20316-01-4), the Stockholm County Council, the Swedish Society of Medicine, HKH Kronprinsessan Lovisas F[ouml]rening f[ouml]r Barnasjukv[aring]rd, S[auml]llskapet Barnav[aring]rd, Stiftelsen Frimurare Barnhuset i Stockholm, Wera Ekstr[ouml]m fond f[ouml]r Pediatrikf[ouml]rskning, Erik and Edith Fernstr[ouml]ms foundation, M[auml]rta och Gunnar V. Philipsons Stiftelse and the Karolinska Institute.[br][br]Nothing to Disclose: SSS, ACA, MC, ON 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1879 305 2225 MON-334 PO04-01 Monday 1894 2012


1890 ENDO12L_MON-335 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Evidence for Leptin-Mediated Bone Mass Regulation Via the Hypothalamus in Humans Using fMRI Pouneh K Fazeli, Elizabeth A Lawson, Laura M Holsen, Hang Lee, Jill M Goldstein, Anne Klibanski Massachusetts General Hospital/Harvard Medical School, Boston, MA; Brigham and Women[apos]s Hospital, Harvard Medical School, Boston, MA; Massachusetts General Hospital, Boston, MA Leptin, a hormone produced by subcutaneous adipose tissue, is a central regulator of bone mass and appetite. In animal models, the bone-regulating actions of leptin are mediated through the ventromedial hypothalamus, and the appetite-regulating actions through the arcuate nucleus. Women with anorexia nervosa (AN) have markedly decreased leptin levels and bone mineral density (BMD) in the setting of low weight, and peripheral leptin levels are positively associated with BMD. Whether the bone mass-regulating actions of leptin are mediated through the hypothalamus in humans is unknown. We hypothesized that the strong association between leptin levels and BMD is mediated by hypothalamic activation in response to appetite stimuli using a novel functional magnetic resonance imaging (fMRI) paradigm. We studied 16 women: 11 with AN (mean age: 22.2 [plusmn] 2.6 (SD) years) and 5 healthy controls (HC) (21.8 [plusmn] 1.3 years). We measured fasting leptin levels, BMD of the lumbar spine, lateral spine and hip by dual energy x-ray absorptiometry. Fasting fMRI was done to measure hypothalamic activation in response to viewing high-calorie foods vs. objects. Women with AN had lower BMI vs. HC (17.6 [plusmn] 1.2 vs. 22.2 [plusmn] 1.6; p=0.002). BMD was lower at the lumbar spine (0.84 [plusmn] 0.09 g/cm[sup]2[/sup] vs. 1.13 [plusmn] 0.05 g/cm[sup]2[/sup]; p[lt]0.0001), lateral spine (0.63 [plusmn] 0.09 g/cm[sup]2[/sup] vs. 0.79 [plusmn] 0.03 g/cm[sup]2[/sup]; p[lt]0.001) and hip (0.79 [plusmn] 0.09 g/cm[sup]2[/sup] vs. 1.02 [plusmn] 0.09 g/cm[sup]2[/sup]; p[lt]0.01) vs. HC. Leptin levels (AN: 2.8 [plusmn] 1.3 vs. 12.1 [plusmn] 4.9; p=0.002) and hypothalamic activation (p=0.04) were lower in AN vs. HC. In the entire group, BMD was associated with leptin levels at the lumbar spine (R=0.70 ([beta]=0.021); p[lt]0.003), lateral spine (R=0.54 ([beta]=0.011); p=0.03) and hip (R=0.64 ([beta]=0.015); p[lt]0.01). After controlling for hypothalamic activation, the relationship between leptin and BMD was no longer significant at the lateral spine ([beta]=0.007; p=0.2) and hip ([beta]=0.01; p=0.2) and attenuated at the lumbar spine ([beta]=0.018; p=0.02), suggesting that hypothalamic activation may mediate the relationship between leptin and BMD at these sites. This attenuation could not be accounted for by collinearity between leptin and hypothalamic activation (R=0.49; p=0.06). We conclude that the association between leptin and bone mass in humans may be mediated through hypothalamic activation in response to appetite stimuli. Future directions will include prospective studies to assess the impact of changes in leptin levels with weight changes on these parameters.[br][br]Nothing to Disclose: PKF, EAL, LMH, HL, JMG, AK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1128 305 2226 MON-335 PO04-01 Monday 1895 2012


1891 ENDO12L_MON-336 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Children with Higher Urinary Calcium Excretion within the Normal Physiologic Range: A Vulnerable Group Whose Bone Health May Benefit from an Increased Fruit [amp] Vegetable Intake Lijie Shi, Lars Libuda, Eckhard Schonau, Lynda Frassetto, Thomas Remer Research Institute of Child Nutrition, Rheinische Friedrich-Wilhelms University Bonn, Dortmund, Germany; University of Cologne, Cologne, Germany; University of California, San Francisco, CA Reduced bone mineral density (BMD) and bone mass have been observed in children with idiopathic hypercalciuria. Whether urinary calcium excretion at the higher end of the normal physiologic range can influence bone health in healthy children independent of dietary intake is unknown. Urinary calcium was quantified in 603 24-h urine samples from 154 healthy children and adolescents who had [ge]3 urine collections and parallel 3-day weighed dietary records during the 4 years preceding proximal forearm bone analyses by peripheral quantitative computed tomography (pQCT). Urinary potential renal acid load (uPRAL) was determined according to urine ionogram by subtracting measured quantitatively important mineral cations from nonbicarbonate anions. Urinary calcium excretion was significantly associated with volumetric (v)BMD (P = 0.04), almost significantly with cortical bone mineral content (BMC) (P = 0.05), but not with cortical cross-sectional area (CSA) (P = 0.09), total CSA (P = 0.3), or Strength-Strain Index (p = 0.8) in the total population sample. Stratified analyses based on the median split of uPRAL showed that calcium excretion was negatively associated with vBMD (p = 0.007), cortical BMC (p = 0.001), and cortical CSA (p = 0.004) in those children with higher uPRALs, but not in those with low uPRALs (p [gt] 0.3). In conclusion, long-term higher calciuria within the physiological range predicts reduced diaphyseal bone mass and bone density particularly in healthy children and adolescents with long-term unfavorable higher dietary acid load, i.e., with lower fruit and vegetable intake.[br][br]Sources of Research Support: DFG, RE 753/7-1.[br][br]Nothing to Disclose: LS, LL, ES, LF, TR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 246 305 2227 MON-336 PO04-01 Monday 1896 2012


1892 ENDO12L_MON-337 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) BMP2 Enhances Osteoblast Phenotypes of Cryopreserved Bone Marrow-Derived Mesenchymal Stem Cells Isolated from Aging Population Mattabhorn Phimphilai, Peraphan Pothachareon, Chanakarn Phornphutkul, Prachya Kongtawelert Chiangmai University, Chiangmai, Thailand; Chiangmai University, Chiangmai, Thailand; Chiangmai University, Chiangmai, Thailand [bold]Introduction: [/bold]Osteoporosis in aging is partly caused by the reduction of osteoblast differentiation and activity which entails the reduction of bone formation and aged-related bone loss. Moreover, the decline in bone formation may lead to a failure of fracture healing in these patients who are at risk for fractures. To promote bone formation, the combination use of mesenchymal stem cells (MSC) and potent osteoinductive factors has been proposed. Cryopreservation of the MSC has been proved for preserving osteogenic differentiation of the cells as well as BMP2, a potent osteoinductive factor, has been shown to stimulate osteogenic differentiation of the MSC. However, the effect of BMP2 in cryopreserved MSC derived from aging population remains further explored.[br][bold]Objective: [/bold]To determine the differentiation toward osteoblast of cryopreserved MSC isolated from aging population with adenovirus-carrying BMP2 gene (AdBMP2).[br][bold]Method:[/bold] Bone marrow from 4 young adults and 8 aging donors were collected. Bone marrow-derived MSC were isolated by plastic adhesion technique, and were characterized by cell surface markers including CD34, CD45, CD90 and CD105 using flow cytometry. The isolated cells were cryopreserved at -80 [sup]o[/sup]C for 3 months. AdBMP2 was used to introduce BMP2 gene into the cells; the BMP2 protein was determined by western blot analysis. The osteoblast specific genes including alkaline phosphatase (ALP), collagen type 1 (COL1) and osteocalcin (OCN) were analyzed by real-time PCR; the mineralization was determined by alizarin-red S staining.[br][bold]Result:[/bold] Mononuclear cells isolated from either young adults or aging donors showed similar cell surface markers, CD105+ and CD90+ but CD34- and CD45-, confirming mesenchymal stem cells (MSC). Cryopreserved MSC maintained the cell surface characters of the MSC. BMP2 protein was detected only in AdBMP2 infected-MSC, indicating the enhancement of BMP2 production in the BMP2-gene transferred MSC. Comparing to control, AdBMP2 enhanced the mRNA level of ALP, COL1 and OCN, as well as the mineralization in similar extent in the MSC isolated either from young or aging donors. In addition, AdBMP2 enhanced osteoblast-specific genes expression and mineralization in cryopreserved MSC derived from aging population in the similar manner to those in the primary culture.[br][bold]Conclusion:[/bold] BMP2 enhances differentiation toward osteoblast of the cryopreserved bone marrow-derived mesenchymal stem cells isolated from aging population.[br][br]Nothing to Disclose: MP, PP, CP, PK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 352 305 2228 MON-337 PO04-01 Monday 1897 2012


1893 ENDO12L_MON-338 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Evidence That Ras Signaling Pathway Promotes Skeletal Healing in a Mouse Model of Tail Vertebra Drill-Hole Subburman Mohan, Jon Wergedal, Hongrun Yu Jerry L Pettis VA Memorial Medical Center, Loma Linda, CA; Loma Linda University, Loma Linda, CA Fracture healing is impaired in 5-10% of patients with seriously delayed union or non-union bone fractures. In order to find effective therapies to promote healing of non-union fractures, it is important to identify the cellular processes and signaling pathways that contribute to regeneration of skeletal tissues. In a previous study, we demonstrated that healing of tail vertebra hole (TVH) varied significantly among different inbred strains of mice, and these differences were largely genetically determined with an estimated heritability of 72%. Using [italic]in vivo[/italic] Micro-CT to accurately quantify the extent of bone healing at different times after drill-hole, we determined in this study that the extent of skeletal healing was several fold greater in the MRL/MpJ mice, a good healer of several soft tissues compared to C57BL/6J mice, a poor healer. Histomorphometric studies revealed that the healing of TVH in MRL/MpJ mice primarily occurs via intramembranous ossification as there was no evidence of cartilage in the drill-hole. Whole genome microarray analysis of healing tissue of MRL/MpJ mice at day 5 and day 10 after drill-hole revealed strong up-regulation of several osteoblast-specific genes (type I collagen, periostin, osterix) compared to day 0 and uninjured vertebra. In addition, we found that a number of genes related to the [underline]ra[/underline]t [underline]s[/underline]arcoma (Ras) signaling pathway were differentially expressed in the healing tissue of MRL/MpJ mice at both day 5 and day 10 after drill-hole compared to day 0. To determine the role for the Ras signaling pathway in rapid healing of TVH, we tested the local effects of an established chemical inhibitor of Ras signaling, S-trans, trans-farnesylthiosalicylic (FTS). FTS was injected subcutaneously into in the drill-hole region of MRL/MpJ mice for 10 days. At day 30, 45% of the drill hole was filled by new bone in the vehicle treated control mice, while only 27% of the drill hole was regenerated in the FTS treated mice. Based on our findings, we conclude that rapid healing of TVH in MRL/MpJ mice occurs primarily via intramembranous bone formation route and is mediated in part via activation of the Ras signaling pathway.[br][br]Nothing to Disclose: SM, JW, HY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2045 305 2229 MON-338 PO04-01 Monday 1898 2012


1894 ENDO12L_MON-339 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Regulation of PC2 and 7B2 Expression during Osteocyte Differentiation Adam N Lick, Matthew Prideaux, Lynda F Bonewald, Iris Lindberg University of Maryland-School of Medicine, Baltimore, MD; University of Missouri at Kansas City-School of Dentistry, Kansas City, MO The neuroendocrine protein 7B2 plays a prominent role in the regulation of prohormone convertase 2 (PC2) activity, an enzyme involved in the production of various peptide hormones. 7B2 interacts with proPC2 to prevent an inactivating aggregation process; conversely, the 7B2 carboxyl-terminal (7B2-CT) peptide inhibits mature PC2 activity. It has been recently discovered that osteoblasts and osteocytes in bone secrete fibroblast growth factor 23 (FGF-23), a protein involved in the regulation of bone mineralization and blood phosphate levels. Recent data from our collaborators indicate that 7B2 and PC2 may contribute to physiological FGF-23 cleavage. In order to investigate this idea in a cell culture model, we utilized a variety of bone cell lines and Western-blotted for PC2 and 7B2 expression under both non-differentiating and differentiating conditions. Only one bone cell line out of the five tested, IDG-SW3, exhibited immunoreactivity for both 7B2 and PC2. While secreted levels of immunoreactive PC2 remained constant during the three weeks of differentiation, secreted 7B2 levels began to diminish after two weeks, when these cells begin to express mature osteocyte markers such as sclerostin. Concurrently with the diminution of secreted 7B2, stored, intracellular proPC2 and 7B2 began to appear in the cells. Interestingly, we also observed immunoreactivity for a high molecular weight PC2 and 7B2-CT immunoreactive (ir) complex in cell extracts at the two-week differentiation time point. The presence of 7B2-CT-ir together with mature PC2-ir in the blots implies active inhibition of enzyme, and correlates with our finding of a total lack of PC2 activity in the secretion medium at this time point. The involvement of PC2 in FGF-23 processing is supported by our observation that recombinant FGF-23 can be efficiently cleaved in vitro by recombinant PC2. These data provide further support for the idea that PC2 can serve as an FGF-23-cleaving enzyme in osteoblasts and osteocytes, and also indicate that the inhibitory C-terminus of 7B2 assists in the control of PC2 activity in differentiating osteocytes.[br][br]Sources of Research Support: NIH Grant DK49703 awarded to IL.[br][br]Nothing to Disclose: ANL, MP, LFB, IL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2166 305 2230 MON-339 PO04-01 Monday 1899 2012


1895 ENDO12L_MON-340 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) 24,25-Dihydroxyvitamin D Promotes the Osteogenic Maturation of Human Mesenchymal Stem Cells through the Regulation of p63 Kevin M Curtis, Kristina K Aenlle, Ketian Chen, Bernard A Roos, Guy A Howard Veterans Affairs Medical Center, Miami, FL; University of Miami Miller School of Medicine, Miami, FL; University of Miami Miller School of Medicine, Miami, FL; University of Miami Miller School of Medicine, Miami, FL; University of Miami Miller School of Medicine, Miami, FL Bone homeostasis and repair are regulated by vitamin D and its metabolites, of which 1[alpha],25-dihydroxyvitamin D (1,25OHD) has been considered the most biologically active and relevant. Vitamin D metabolites include 24,25-dihydroxyvitamin D (24,25OHD), a major metabolite with heretofore unappreciated activity. We present here results showing that 24,25OHD regulates human mesenchymal stem cell (hMSC) osteogenic maturation.[br]Our data establishes that 24,25OHD inhibits hMSC proliferation, increases alkaline phosphatase activity and mineralization, and decreases 1[alpha]-hydroxylase expression, potentially decreasing the ability to convert 25OHD to 1,25OHD. In addition, 24,25OHD but not 1,25OHD induced mineralization in the absence of dexamethasone -- commonly used for osteogenic induction and known to up-regulate the VDR. This data supports a role for 24,25OHD during osteogenic maturation, suggesting that previously published studies using dexamethasone are skewed toward a 1,25OHD response.[br]Currently, we don[apos]t understand how 24,25OHD and 1,25OHD differentially mediate this process. During murine development, the transcription factor p63 is required for skeletal formation (1). In addition, our work (2) demonstrates that p63 is integral for 1,25OHD receptor, VDR expression in hMSC. Therefore, since p63 regulates 1,25OHD/VDR, the question is raised as to its regulation of 24,25OHD actions and overall osteogenic maturation. p63 gene expression is complex, leading to the formation of TA- and [Delta]Np63 isoforms, and three splice variants ([alpha],[beta],[gamma]). The p63 gene products can activate or repress growth and differentiation, however their role(s) during osteogenic maturation are not established.[br]Our studies demonstrate that during the progression of immature hMSC toward the osteogenic phenotype, there is a switch from TAp63[alpha],[beta] to TAp63[gamma] expression. The addition of 1,25OHD up-regulated the [Delta]Np63 isoforms, while 24,25OHD increased TA/[Delta]Np63[gamma] mRNA expression and TAp63[alpha] protein expression. These results link 1,25OHD up-regulation of [Delta]Np63, required for terminal differentiation (3), and 24,25OHD up-regulation of p63[gamma], known to regulate VDR expression (4). These results support a generalized paradigm implicating p63 as a key player during hMSC osteogenic maturation and possibly bone repair in vivo. We hypothesize that the actions of vitamin D on osteogenic maturation and bone repair are due to a regulatory relationship between p63 gene products and unique 24,25OHD/1,25OHD effects.[br][br](1) Yang A, Schweitzer R, Sun D, et al., p63 is essential for regenerative proliferation in limb, craniofacial and epithelial development. Nature, 1999. 398(6729): p. 714-8 (PMCID: 10227294). (2) Chen K, Roos BA, Howard GA: Hepatocyte Growth Factor (HGF) and 1,25-dihydroxyvitamin D Together Stimulate Human Bone Marrow-Derived Stem Cells Toward the Osteogenic Phenotype by HGF-induced Up-Regulation of VDR. BONE 2012 (Under Revision). (3) Yang A, Kaghad M, Wang Y, et al., p63, a p53 homolog at 3q27-29, encodes multiple products with transactivating, death-inducing, and dominant-negative activities. Mol Cell, 1998. 2(3): p. 305-16 (PMCID: 9774969). (4) Kommagani R., Caserta TM., and Kadakia MP. Identification of vitamin D receptor as a target of p63. Oncogene (2006) 25, 3745-3751.[br][br]Sources of Research Support: Veterans Affairs Merit Review Awards (GH, BR).[br][br]Nothing to Disclose: KMC, KKA, KC, BAR, GAH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 685 305 2231 MON-340 PO04-01 Monday 1900 2012


1896 ENDO12L_MON-341 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Genetics of Familial Osteoporosis and Idiopathic Hypercalciuria Katherine Tighe Stanley, Kevin Le, Murray Favus University of Chicago, Chicago, IL; University of Chicago, Chicago, IL Family history plays a major role in the development of osteoporosis (OP) and idiopathic hypercalciuria (IH). Bone mineral density (BMD) is highly heritable, contributing 57 to 92% of peak BMD in premenopausal twin pairs. Familial osteoporosis (FOP) not associated with aging or menopause and without a specific etiology has been described. Although population-based GWAS or candidate gene association studies have defined multiple genes with small contributions to BMD, FOP likely involves different genes with larger effects. IH may also cause low BMD and increased fracture risk but differs from OP by pathophysiology and treatment. A few population-based candidate gene association studies have failed to show conclusive associations with IH. Most studies using genetic associations with kidney stones as a marker of hypercalciuria leave out many subjects with IH and low bone density but without stone formation. We hypothesize that the familial forms of IH and FOP result from mutations in the protein coding regions in a limited number of genes. The study aim is to identify genetic variants associated with OP or IH in families using exome sequencing. Study subjects are selected along with their families from the University of Chicago Bone Clinic based on FOP or IH and family history of low BMD with or without IH. DNA is isolated for exome sequencing. Subjects are phenotyped using BMD, 24 hour urine Ca excretion, and other pertinent blood and urine testing. Nine subjects from two families have been studied to date. Probands have either OP or hypercalciuria. One family has a proband with osteoporosis and hypercalciuria and several members with low BMD as well as one with kidney stones and another who is now known to be hypercalciuric. In this family, the proband has a femoral neck T score of -2.9 and 24 urine calcium excretion of 245 mg calcium per gram urine creatinine (normal [lt]140). Her 3 sisters have T scores ranging from -1.1 to -2.1. Several other family members have had fractures and await BMD testing. Several subjects in the second family have low BMD and hypercalciuria. In this family, the proband is a premenopausal woman with a Z score of -2.1 and 24 hour urine calcium of 401 mg Ca/24 hours (normal[lt] 250). Her mother and twin sons all have low bone mass with Z scores of -2.1-3.1. In conclusion, low bone density and hypercalciuria are sufficiently common among family members to permit exome sequencing to determine genetic associations with these traits.[br][br]Disclosures: MF: Consultant, CVS/Caremark. Nothing to Disclose: KTS, KL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1761 305 2232 MON-341 PO04-01 Monday 1901 2012


1897 ENDO12L_MON-342 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Identification of Peptides That Bind Mouse Epiphyseal Chondrocytes by Phage Display Crystal Sao Fong Cheung, Julian Chun Kin Lui, Jeffrey Baron National Institute of Child Health and Human Development, Bethesda, MD Growth plate cartilage is susceptible to a wide variety of genetic, endocrine, metabolic, and iatrogenic disorders that can impair bone growth and cause bone deformity. To develop improved experimental, diagnostic, and therapeutic approaches, we sought to identify peptides with high binding affinity for growth plate cartilage.[br]We used a phage display library, in which 12-amino acid peptide variants encoded by random DNA sequences are expressed and displayed at the N-terminus of the minor coat protein pIII of bacteriophage. The phage library was incubated with cultured epiphyseal chondrocytes isolated from 6-days-old mice. After repeated washing, bound phage were eluted, amplified, and taken through two additional binding/amplification cycles to enrich the pool in favor of cartilage-binding sequences. Twenty phage clones were randomly picked, and their ability to bind cultured chondrocytes was assessed by ELISA. Multiple phage clones demonstrated increased affinity to cultured chondrocytes, when compared to a phage lacking the peptide-encoding insert (insertless phage). For three of these clones, we performed confirmatory test of cartilage-binding affinity. Each of these selected phage was mixed in equal amounts with the insertless phage, and the mixture was incubated with cultured chondrocytes, washed, and eluted. DNA from eluted phage was PCR-amplified and subjected to gel electrophoresis. Compared to the input mixture, the output exhibited an increased ratio of insert-containing to insertless DNA band intensity, indicating inserted phage was better at binding chondrocytes. The selected phage did not show any preferential binding to cultured adult mouse hepatocytes in comparison to insertless phage, suggesting a tissue-specific interaction. DNA sequencing revealed that two of the peptides have substantial amino acid sequence similarity. Further investigation is required to determine the peptide binding affinity, tissue specificity, and molecular targets.[br][br]Nothing to Disclose: CSFC, JCKL, JB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 997 305 2233 MON-342 PO04-01 Monday 1902 2012


1898 ENDO12L_MON-343 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Des-A Analogs of 1[alpha],25-Dihydroxy-19-Norvitamin D3 Agnieszka Glebocka, Lori A Plum, Hector L DeLuca University of Wisconsin, Madison, WI The steroid hormone 1[alpha],25-dihydroxyvitamin D3 [1[alpha],25-(OH)2D3]is the active metabolite of vitamin D3 which exerts its control over a multitude of biological processes related to calcium and phosphate homeostasis, cell proliferation and differentiation, and immune regulation. Unfortunately, the therapeutic application of 1[alpha],25-(OH)2D3 is limited by induction of hypercalcemia. The need for vitamin D analogues with more selective biological profiles has led to the synthesis of more than three thousand analogues of 1[alpha],25-(OH)2D3. Most of these compounds have structural modifications in the side chain and A-ring; there is also increasing number of modifications in the CD-rings and limited number in triene system. Herein, we report the synthesis and biological evaluation of Des-A analogs of 1[alpha],25-dihydroxy-19-norvitamin D3, developed to study the role of A-ring in the biological activity of 1,25-(OH)2D3. Interestingly, few of these new, nonsteroidal compounds show substantial binding to Vitamin D Receptor and one of them is characterized by very selective activity on intestinal Ca absorption.[br][br]Nothing to Disclose: AG, LAP, HLD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2029 305 2234 MON-343 PO04-01 Monday 1903 2012


1899 ENDO12L_MON-344 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) 1-Desoxy and 3-Desoxy Analogs of 2MD and Its 20R-Epimer: Synthesis and Biological Activity Izabela K Sibilska, Rafal R Sicinski, Lori L Plum, Hector F DeLuca University of Warsaw, Warsaw, Poland; University of Wisconsin, Madison, WI During our ongoing structure-activity studies in the vitamin D area we obtained (20S)-1[alpha],25-dihydroxy-2-methylene-19-norvitamin D3. This analog, called 2MD, is characterized by significantly enhanced calcemic activity and is currently evaluated as a potential drug for osteoporosis. Therefore, it was of interest to synthesize also its 1-desoxy and 3-desoxy analogs and examine their biological action. The aim of these studies was not only to evaluate a role of the 3-hydroxyl group but also to solve an intriguing problem: can 1-desoxy vitamin be hydroxylated in vivo in the allylic C-1 position despite the lack of the exomethylene moiety at C-10? A new synthetic path has been elaborated leading to the desired A-ring phosphine oxide synthons that started from the commercially available achiral compound, 1,4-cyclohexanedione monoethylene acetal. The Wittig-Horner reaction of the known protected 25-hydroxy Grundmann ketone and the corresponding phosphine oxides gave 3-desoxy-2-methylene-19-nor-1[alpha],25-(OH)2D3 and 2-methylene-19-nor-25-OH-D3. Analogous coupling of (20S)-25-hydroxy Grundmann ketone provided the title compounds. The biological activity of the synthesized vitamin D analogs was evaluated.[br][br]Nothing to Disclose: IKS, RRS, LLP, HFD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 767 305 2235 MON-344 PO04-01 Monday 1904 2012


1900 ENDO12L_MON-345 POSTER SESSION: Bone Biology [amp] Metabolic Bone Disorders (1:30 PM-3:30 PM) Estrogen Facilitates Osteoblast Differentiation by Upregulating BMP-4 Signaling Fumio Otsuka, Yoshinori Matsumoto, Mariko Takano-Narazaki, Tomoko Miyoshi, Kenichi Inagaki, Kanako Ogura-Ochi, Hirofumi Makino Okayama University Graduate School, Okayama, Japan Estrogens are key factors for determining bone mass in postmenopausal women by maintaining balance of osteoblastic bone formation and osteoclastic bone resorption. Recent studies have shown impaired proliferation and osteogenic activity of mesenchymal stem cells after estrogen depletion. Estrogen administration enhances bone formation in vivo. These findings suggest that estrogen also facilitates osteogenesis in addition to its protective effect by suppressing osteoclast-induced bone resorption. However, the precise mechanism of estrogen-induced bone formation remains unclear. Here we investigated the cellular mechanism by which estrogen interacts in the process of osteoblastic differentiation regulated by BMP-4 using mouse MC3T3-E1 cells that express estrogen receptors (ER) and BMP-4. The osteoblastic cell line MC3T3-E1 established from a C57BL/6 mouse calvarium has the capacity to differentiate into osteoblasts and osteocytes, resulting in calcified bone tissue in vitro. MC3T3-E1 cells express BMP-4, the expression of which decreases during normal differentiation. In the present study, it was revealed that estradiol enhanced BMP-4-induced Runx2, osterix, ALP and osteocalcin expression in MC3T3-E1 cells. BMP-4-induced mineralization shown by Arizalin red staining was also facilitated by estrogen treatment. It was also found that estrogen upregulated BMP-4-induced Smad1/5/8 phosphorylation, BRE-Luc activity and Id-1 mRNA expression. The expression of BMPRII was increased by estrogen and that of inhibitory Smad6/7 was decreased by estrogen action in MC3T3-E1 cells. Of note, the enhanced expression of osterix, ALP and osteocalcin mRNAs induced by BMP-4 and estrogen was reversed in the presence of an ER antagonist. Given that membrane-impermeable estrogen also upregulated BMP-4-induced expression of osteoblastic markers, non-genomic ER activity is involved in the mechanism by which estrogen enhances BMP-4-induced osteoblast differentiation in MC3T3-E1 cells. On the other hand, the expression of ER[alpha] and endogenous BMP-4 was suppressed by BMP-4 treatment regardless of the presence of estrogen, implying the presence of a negative feedback loop for osteoblast differentiation. Collectively, estrogen is functionally involved in the process of osteoblast differentiation regulated by BMP-4 through upregulating BMP sensitivity of MC3T3-E1 cells.[br][br]Nothing to Disclose: FO, YM, MT-N, TM, KI, KO-O, HM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1483 305 2236 MON-345 PO04-01 Monday 1905 2012


1901 ENDO12L_MON-346 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Prevalence and Associations of Vitamin D Deficiency in Medical Outpatients and Inpatients Marianne Ghobrial, Katayoun Edalat-Parsi, Evelina Svrdlan, Ivance Pugoy, Abhishek Sawant, Paul K Mills, Soe Naing UCSF Fresno Center for Medical Education and Research, Fresno, CA; Community Regional Medical Center, Fresno, CA; Fresno Medical Education Program, University of California, San Francisco, Fresno, CA Aim:[br]To determine the prevalence of vitamin D deficiency (VDD) and associations between VDD and clinical parameters in medical outpatients (OP) and inpatients (IP)[br]Method:[br]This retrospective study was conducted at a community hospital in Fresno, California. The medical OP and IP, who had serum 25 hydroxyvitamin D [25(OH)D] measured from July 2009 to June 2011 for any reason, were included in the study.[br]Results:[br]We studied a total of 1548 patients: 536 OP (mean age 55years, 62% female, 55% Hispanics, mean 25(OH)D 19.4mg/dl) and 862 IP (mean age 59years, 49% female, 41% Hispanics, mean 25(OH)D 16.3mg/dl). 16.4% of OP and 30.4% of IP had severe VDD ([lt]10mg/dl) whereas 56.6% OP and 70.1% IP had VDD ([lt]20). In both groups, those with VDD were younger (OP 54 vs 57years;IP 58 vs 63), had higher body weight (OP 188 vs 181lbs;IP 175 vs 162), lower total calcium (OP 9.3 vs 9.5mg/dl;IP 8.5 vs 8.8), higher HbA1c (OP 7.2 vs 6.6%;IP 6.8 vs 6.0), lower hemoglobin (Hb) (OP 12.2 vs 12.8mg/dl;IP 10.3 vs 10.8), higher triglyceride (OP 170 vs 135mg/dl;IP 139 vs 121), higher LDL (OP 103 vs 93mg/dl;IP 82 vs 75), higher TSH (OP 5.5 vs 2.1mU/L;IP 4.4 vs 3.8), higher PTH (OP 129 vs 113mg/dl;IP 219 vs 141), than those without VDD. In multivariate regression analysis, the presence of DM (OR 4.25; 95% CI 1.46-12.39), higher LDL (OR 1.025; 95% CI 1.00-1.04) and lower Hb (OR 0.61; 95% CI 0.44-0.85) levels in OP and younger age (OR 0.969; 95% CI 0.945-0.994), lower albumin (OR 0.374; CI 0.198-0.708) and higher PTH (OR 1.003; CI 1.000-1.006) levels in IP were independently associated with VDD.[br]Discussion:[br]Prevalence of VDD is surprisingly high though there is abundant sunlight in Fresno at Central California (36.7[deg] N latitude and 267 total days with sun). Old age is considered to be one of the major risk factors for VDD in general population. However, in our study, those with VDD were significantly younger than those without. The prevalence of VDD was much higher and mean 25(OH)D level was significantly lower in IP than in OP, that may suggest that acute medical conditions or exacerbation of chronic diseases may associate with higher risk of VDD.[br]Conclusion:[br]VDD was highly prevalent in both medical OP and IP in this sunny region. Prevalence of severe VDD was about 2 times higher in IP than in OP. The presence of DM, elevated LDL and low Hb levels were significantly associated with VDD in OP whereas younger age, lower albumin and higher PTH levels were independently correlated with VDD in IP.[br][br]Nothing to Disclose: MG, KE-P, ES, IP, AS, PKM, SN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1173 306 2237 MON-346 PO07-01 Monday 1906 2012


1902 ENDO12L_MON-347 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Vitamin D Deficiency in Middle East Nishanth Sanalkumar, Priya Sreekumaran Nair, Srinivas Bontha Al Zahra Hospital, Sharjah, United Arab Emirates; Dr Sunny Medical Center, Sharjah, United Arab Emirates Despite ample sunshine,Vitamin D deficiency is reported from several countries in middle east(1). We describe the prevalence of vitamin D deficiency in patients who were seen in the outpatient clinics of Al Zahra Hospital, Sharjah, UAE over a period of 14 months.[br]Methods: We analyzed the demographic profile of all patients who had 25-hydroxy vitamin D (25[OH]D) levels tested at the Al Zahra hospital, Sharjah from January 2010 to February 2011.Severity of vitamin D deficiency was determined based on the serum 25[OH]D level.Total 25[OH]D level in serum was measured using Diasorin chemiluminescence assay (normal range 75-200 nmol/L).Vitamin D deficiency was defined as 25[OH]D levels below 50 nmol/L (20 ng/mL). Levels between 50-75 nmol/L (20-30 ng/mL)was taken as vitamin D insufficiency and above 75 nmol/L (30 ng/mL), normal. Patients with 25[OH]D levels below 25 nmol/L (10 ng/mL) were considered to have severe vitamin D deficiency.The mean 25[OH]D levels in different nationalities, age groups and in men and women were compared.Statistical analysis was done using SPSS 10.0 statistical software.[br]Results: Serum 25[OH]D levels were available for 3449 patients. Mean age([plusmn]SD) of the population tested was 38.6 [plusmn]13.4 years. Male to female ratio was 1:3.More than 60 nationalities were represented and were divided into 5 groups [ndash] UAE nationals, Non UAE Arabs, Caucasians, South Asians, Others.The mean ([plusmn]SD) 25[OH]D level was 30.34 [plusmn] 18.09 nmol/L. Only 111 patients (3.2%)had normal vitamin D levels. Vitamin D insufficiency was present in 399 patients (11.6%) and deficiency in 2939 patients (85.2%). Nearly half of all the patients tested had severe vitamin D deficiency (n 1629; 47.2%).The 25 [OH]D level was low in both sexes, but it was significantly lower in females compared with males (29.5[plusmn]18.3 vs 33.5[plusmn]16.9 nmol/L).Mean 25[OH]D level was below normal in all nationalities, but was lowest in UAE nationals (27.29nmol/L) and highest in Caucasians (39.97 nmol/L).Vitamin D level was low across all ages. It was relatively better in the first decade (45.8nmol/L).Mean 25[OH]D levels were higher in January to June period compared with July-December.[br]Conclusion: Vitamin D deficiency, including severe vitamin D deficiency, is very common in Sharjah.Daily supplementation with 400-800 IU of vitamin D is recommended for prophylaxis(2), but higher doses may be needed to keep the 25[OH]D levels above 75 nmol/L.[br][br](1)The Middle East and Africa Regional Audit. The Epidemiology, cost and burden of Osteoporosis in 2011. www.iofbonehealth.org. (2)Holick MF, Binkley NC, Heike A. Evaluation, Treatment and Prevention of Vitamin D deficiency. J Clin Endocrinol Metab July 2011, 96(7) 1911-30.[br][br]Nothing to Disclose: NS, PSN, SB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 677 306 2238 MON-347 PO07-01 Monday 1907 2012


1903 ENDO12L_MON-348 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Maternal Vitamin D Status during the Course of Pregnancy in Thai Women Natthinee Charatcharoenwitthaya, Tongta Nanthakomon, Charintip Somprasit, Athita Chanthasenanont, La-or Chailurkit, Boonsong Ongphiphadhanakul Faculty of Medicine, Thammasat University, Pathumthani, Thailand; Faculty of Medicine, Thammasat University, Pathumthani, Thailand; Ramathibodi Hospital, Mahidol University, Bangkok, Thailand [bold]Background[/bold] Increasing evidence suggests that attaining optimal vitamin D status during pregnancy may decrease adverse pregnancy outcomes as well as susceptibility to many chronic diseases in the offsprings. Although Thailand is sunshine abundant, vitamin D inadequacy is common. The objectives of the present study are to evaluate the prevalence of vitamin D inadequacy, and its predictive factors, during the course of pregnancy in Thai women. Pregnancy outcomes in relation to vitamin D status were also assessed.[br][bold]Methods[/bold] A total of 120 pregnant women were enrolled at the first antenatal visit. The inclusion criteria were age 18-40 years, gestational age less than 14 weeks, and no disease known to affect calcium and vitamin D metabolism. Clinical data and blood samples were obtained 3 times, i.e., at the first visit, in the second trimester and in the third trimester of pregnancy. The 25 hydroxyvitamin D (25OHD) concentrations were measured using liquid chromatography tandem mass spectrometry. Vitamin D inadequacy and deficiency were defined as the levels of 25OHD [lt] 30 and [lt] 20 ng/mL, respectively.[br][bold]Results[/bold] The prevalence of vitamin D inadequacy was 83.3% in the first trimester and significantly decreased to 30.9% and 27.4% in the subsequent trimesters (p[lt]0.0001). The significant predictors of vitamin D inadequacy were not taking prenatal vitamin (OR=10.67; p[lt]0.0001) and not taking vitamin fortified milk (OR 2.74; p[lt] 0.001). Intake of at least 400 IU/d of vitamin D from prenatal vitamin significantly increased the 25OHD levels by 12.3[plusmn]6.5 ng/mL (p[lt]0.0001) from baseline and could prevent vitamin D deficiency in the 2[sup]nd[/sup] and 3[sup]rd[/sup] trimesters. The risk of vitamin D inadequacy was significantly lower in women taking [ge]400 IU/d than women taking [lt]400 IU/d of vitamin D from prenatal vitamin (OR 0.22; p[lt]0.01). No differences in the 25OHD levels and prevalence of vitamin D inadequacy were found between women taking 400 IU/d and 800 IU/d of vitamin D from prenatal vitamin (p=0.20). There were no associations between vitamin D inadequacy and adverse pregnancy outcomes.[br][bold]Conclusions[/bold] Vitamin D inadequacy is highly prevalent in pregnant Thai women especially in the first trimester. Supplementation of prenatal vitamin prior conception and during pregnancy is recommended. The dose of vitamin D at 400 IU/d is high enough to prevent vitamin D deficiency but inadequate to prevent vitamin D inadequacy even at 800 IU/d in pregnant Thai women.[br][br]Nothing to Disclose: NC, TN, CS, AC, L-oC, BO 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 357 306 2239 MON-348 PO07-01 Monday 1908 2012


1904 ENDO12L_MON-349 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Prevalence of Vitamin D Deficiency and Associated Factors in Turkey Ilhan Satman, Nese Colak Ozbey, Harika Boztepe, Sibel Kalaca, Sema Genc, Bulent Canbaz, Beyhan Omer, Faruk Alagol, TURDEP-II Study Group Istanbul University, Istanbul Faculty of Medicine, Istanbul, Turkey; Marmara University Faculty of Medicine, Istanbul, Turkey; Istanbul University, Istanbul Faculty of Medicine, Istanbul, Turkey Aim: Serum 25OH Vit-D (25OHD) is considered to be the best indicator of overall Vit-D status of an individual. However, the threshold level of Vit-D deficiency may differ, and its determinants are less clear among populations. The objective of this study was to determine the prevalence of Vit-D deficiency, to examine the factors to affect having reasonable level of 25OHD, and to define if there is a population-specific threshold level indicating Vit-D deficiency.[br]Methods: We used data from recently completed, population-based, cross-sectional TURDEP-II survey; including 9,154 adult Turkish people (mean[plusmn]SD; 45.3[plusmn]15.4 yrs., 65% female). The study was conducted in randomly assigned 540 urban/rural centers in Turkey from Jan 15 to June 11, 2010.[br]Results: Among female, mean 25OHD was lower, and mean PTH was higher than males (25OHD; 8.9[plusmn]6.8 vs 12.1[plusmn]7.7 ng/mL, PTH 40.0[plusmn]24.9 vs 33.6[plusmn]17.1 pg/mL, p[lt]0.001). Approximately 60% of the population had severe/moderate Vit-D deficiency (1). From winter to summer, mean 25OHD increased from 9.6[plusmn]6.9 to 12.5[plusmn]8.5 ng/mL (p[lt]0.001), the rate of increase was more remarkable among male (29% vs 45%). Mean PTH inversely correlated with 25OHD (r=-0.146, p[lt]0.001). The optimum cut-off level of 25OHD at suppressed PTH (mean-2SD: [le]25 pg/mL) was defined as 7.8 ng/mL with 65% sensitivity and 50% specificity by ROC analysis. Multiple logistic regression model indicated that male gender, total chol. and IGF-1 were positively; in contrast eGFR and PTH were inversely associated with having 25OHD [ge]20 ng/mL. A separate analysis in female revealed that age, and central obesity were positive factors; whereas less than formal education, current/past smoking, and past alcohol use were inverse factors significantly associated with having reasonable 25OHD. Similarly, male gender, triglycerides, total chol, FT4, 25OHD, IGFBP3 and hs-CRP were positive; in contrast age, and BMI were negative factors significantly associated with suppressed PTH. In female parity was a positive factor; whereas age, current/past smoking, and known diabetes were negative factors significantly associated with having suppressed PTH.[br]Conclusion: The prevalence of Vit-D deficiency was remarkably common in the adult, particularly female population of Turkey. Lifestyle and general health factors may be related with having reasonable 25OHD and/or suppressed PTH levels. The impact of Vit-D deficiency on general health and on chronic disease management needs to be further evaluated.[br][br]The Endocrine Society. J Clin Endocrinol Metab 2011; 96:1911.[br][br]Sources of Research Support: 1. The Society of Endocrinology and Metabolism, Turkey (SEMT). 2. Istanbul University Scientific Research Fund.[br][br]Nothing to Disclose: IS, NCO, HB, SK, SG, BC, BO, FA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1224 306 2240 MON-349 PO07-01 Monday 1909 2012


1905 ENDO12L_MON-350 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Evaluation of Response to Treatment with 50,000 IU of Vitamin D3 in Deficient Patients Tatiane Vilaca, Marilia Camargo, Marise Lazaretti-Castro Universidade Federal de S[atilde]o Paulo - UNIFESP, S[atilde]o Paulo, Brazil Background: Inadequate vitamin D status has been associated with an increased risk of several diseases, including osteoporosis, cardiovascular disease, diabetes, cancer, multiple sclerosis and infections. Serum 25-hydroxyvitamin D is the classic marker of vitamin D status and is determined by environmental (season, latitude, sunlight, diet), demographic (ethnicity, body mass index) and genetic (polymorphism in metabolism and transport genes) factors. (1) However, the features that modify the response to vitamin D supplementation remain unclear. The Endocrine Society suggests the use of 50,000 IU of vitamin D2 or D3 weekly over eight weeks for the treatment of deficient adults to achieve a blood level of 25-hydroxyvitamin D above 30 ng/mL. (2)[br]Objective: The aim of the present study was to evaluate the response to supplementation with 50,000 IU of vitamin D3 in deficient patients.[br]Patients and Methods: Twenty-five osteoporotic female patients with vitamin D deficiency (25-hydroxyvitamin D [lt] 20 ng/ml) received eight capsules of 50,000 IU of vitamin D3 for treatment. All capsules were manufactured at the same place. The patients were instructed to take one capsule a week for eight weeks. All patients reported appropriate adherence to treatment and none reported any gastrointestinal problems.[br]Results: After treatment, serum levels of 25-hydroxyvitamin D were reevaluated. More than half of the patients (13/25) did not achieve the levels recommended by the Endocrine Society ([gt] 30 ng/ml). Both groups of patients (successful treatment and unsuccessful treatment) had similar body mass index values and initial pretreatment levels of 25(OH)-vitamin D. The group with the worse response was older.[br]Discussion: A number of authors have reported considerable inter-individual variation in serum 25-hydroxyvitamin D response to supplementation with identical doses. In the present study, this variation was found using the dose recommended by the Endocrine Society. The data suggest that the standard dose may not be enough to treat all patients and that reevaluation of vitamin D levels after treatment should always be considered.[br][br]1 Mason RS et al. European Journal of Clinical Nutrition (2011) 65, 986[ndash]993. 2 Holick MF el al. J Clin Endocrinol Metab. 2011 Jul;96(7):1911-30.[br][br]Sources of Research Support: S[atilde]o Paulo Research Foundation (FAPESP 2008/555677).[br][br]Nothing to Disclose: TV, MC, ML-C 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1229 306 2241 MON-350 PO07-01 Monday 1910 2012


1906 ENDO12L_MON-351 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Gender-Dependent Skeletal Effects of Vitamin D Deficiency in Young Generation Jung Soo Lim, Kyoung Min Kim, Yumie Rhee, Sung-Kil Lim Yonsei University College of Medicine, Seoul, Korea; Yonsei University College of Medicine, Seoul, Korea; Yonsei University College of Medicine, Seoul, Korea [bold]Context: [/bold]The major health threats caused by vitamin D deficiency in young generation have not been fully elucidated.[br][bold]Objective: [/bold]To investigate skeletal and nonskeletal effects of vitamin D deficiency and study the optimal level of serum 25(OH)D in young people.[br][bold]Design and Setting: [/bold]The Fourth Korea National Health and Nutrition Examination Surveys (KNHANES IV) conducted in 2008-9.[br][bold]Participants: [/bold]4276 people (1926 men and 2350 women) aged 10[ndash]40 yrs selected 16 administrative districts of South Korea.[br][bold]Main outcome measures:[/bold] Age-specific changes in bone mineral density (BMD) according to serum 25(OH)D.[br][bold]Results: [/bold]Serum 25(OH)D was less than 25 nmol/l in 18.8% of participants, 25 to [lt]50 nmol/l in 50.0%, 50 to [lt]75 nmol/l in 27.0%, and 75 nmol/l or greater in 4.2%. Vitamin D deficiency was more frequent in women than in men. There were gender differences in the skeletal effects of vitamin D deficiency. In men between 10[ndash]22 yrs old, BMD was significantly higher in the vitamin D sufficient group, and in men between 23[ndash]40 yrs old, a positive correlation between serum 25(OH)D and BMD was observed. However, in women, we could not find significant differences in BMD according to vitamin D level. Vitamin D deficiency in young generation had no remarkable effects on most nonskeletal parameters or on the prevalence of concomitant diseases except rheumatoid arthritis.[br][bold]Conclusions: [/bold]Vitamin D plays an essential role in skeletal health in young people. Moreover, the presence of gender-dependent skeletal effects is an important observation of this study. Reassurance of serum 25(OH)D up to 20[ndash]30 ng/ml or higher is necessary, especially during the modeling phase in men.[br][br]Sources of Research Support: This work was supported by a National Research Foundation of Korea (NRF) grant funded by the Korean government (MEST) (No.20110001024).[br][br]Nothing to Disclose: JSL, KMK, YR, S-KL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1167 306 2242 MON-351 PO07-01 Monday 1911 2012


1907 ENDO12L_MON-352 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Vitamin D Status of Upper Socioeconomic Status Subjects of South Indians Living in High Altitude (Bengaluru) Harinarayan Venkata Chittari, Shalini Joshi, Leena Appicatlaa, Nalini Arunachalam Bangalore Fortis Hospital, Bangalore, India [bold]Background: [/bold]Vitamin D status of south Indian population living in Tirupati (altitude 300 feet above mean sea level) Andhra Pradesh has been documented [sup]1,2[/sup]. Subjects living in near about the same latitude but higher altitude (Bangalore) have not been documented.[br][bold]Aim:[/bold] To study the vitamin D status in healthy upper socioeconomic subjects living in Bangalore, South India (altitude 3000 feet above mean sea level).[br][bold]Methods:[/bold] Healthy males (n=150) and Females (n=606) were studied for their serum albumin, creatinine, calcium (S.Ca), phosphorous (S.Phos), alkaline phosphatase (SAP), 25 OH vitamin D and ntact-PTH (PTH) status.[br][bold]Results:[/bold] (values are mean [plusmn] SEM). Age of males and females were 50[plusmn]1.44; 51[plusmn]0.6 yrs. All subjects had S.Ca (mg/dl), S.Phos (mg/dl), and SAP (IU/L) in the normal range: Males 9.21[plusmn]0.05; 3.67[plusmn]0.05; 92.65[plusmn]5.01 and females 9.16[plusmn]0.02; 3.74[plusmn]0.02; 86.79[plusmn]1.34 respectively. All subjects had Vitamin D deficiency. Vitamin D (ng/ml) levels were 12.69[plusmn]0.55 in males and 13.72[plusmn]0.38 females and PTH(pg/ml) 45.11[plusmn]1.67 males and 50.71[plusmn]0.97 in females. PTH correlated positively with SAP (r = 0.15;P[lt]0.04); negatively with vitamin D (r = - 0.3;P[lt]0.001), S.Cal (r = - 0.12;P[lt]0.02); S.phos (r = - 0.22;P [lt]0. 001). There was a positive correlation between age and vitamin D (0.2;P[lt]0.001); S.Albumin with S.Cal (r=0.3;P[lt]0.001).Whole group was categorized into vitamin D deficiency ([lt]20); insufficiency (20-30); and normal ([gt]30) and analyzed. Vitamin D deficiency, insufficiency and sufficiency (mean [plusmn] SEM in brackets) were found in 86 (10.72[plusmn]0.41), 13.3(24.32[plusmn]0.72) and 0.67(34.10) percent of males and 78.6 (9.88[plusmn]0.23), 14.85(23.48[plusmn]0.28) and 6.55 (37.36[plusmn]1.30) percent of females. The corresponding PTH values (pg/ml) in Vitamin D deficiency, insufficiency and sufficiency were 45.78[plusmn]1.85; 41.11[plusmn]3.81 and 39.00 in males and 53.21[plusmn]1.10; 43.14[plusmn]2.49 and 37.78[plusmn]2.52 in females. There was statistically significant difference (P[lt]0.05) between the groups.[br][bold]Conclusions: [/bold]This study documents high prevalence of hypovitaminosis D in south Indian urban population with upper socioeconomic status living in high altitude. Altitude did not have any significant additional effect on the prevalence of vitamin D status in this study group. Hypovitaminosis D presents as osteopenia and can accelerate osteoporosis.[br][br]1. Harinarayan CV, Ramalakshmi T, Prasad UV, Sudhakar D. Vitamin D status in Andhra Pradesh: a population based study. Indian J Med Res. 2008;127(3):211-8. 2. Harinarayan CV, Ramalakshmi T, Prasad UV, Sudhakar D, Srinivasarao PVLN, Sarma KVS, Kumar EGTV. High prevalence of low dietary calcium, high phytate consumption and vitamin D deficiency in healthy south Indians. Am J Clin Nutr 2007;85:1062-67.[br][br]Nothing to Disclose: HVC, SJ, LA, NAB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1491 306 2243 MON-352 PO07-01 Monday 1912 2012


1908 ENDO12L_MON-353 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Unexpected Low Levels of 25-Hydroxyvitamin D in a Subtropical Area of Brazil: Analysis of 59,812 Samples Rosa Paula Mello Biscolla, Cynthia Brandao, Odete Hirata Nakamura, Milena Teles, Jose de Sa, Valdemir Malechco Carvalho, Jose GH Vieira Grupo Fleury, S[atilde]o Paulo, Brazil [bold]Introduction: [/bold]Vitamin D deficiency and insufficiency have been defined as a 25(OH)D of less than 20 ng/mL and of 21-29 ng/mL, respectively (1). In the literature the prevalence of 25(OH)D deficiency and insufficiency varies among the studies from 20-100%. In Brazil, mainly a tropical country, two studies carried out in the city of Sao Paulo (23o 34[apos]latitude, a subtropical region) had shown a prevalence of low mean levels of 25-hydroxyvitamin D, 25(OH)D, in elderly people (11ng/mL) in spring and in young resident physicians specially in wintertime (24.4 ng/mL) (2,3). [bold]Objective: [/bold]The aim of the study was to evaluate vitamin D status in a population living in a subtropical area of Brazil. [bold]Patients and Methods: [/bold]Records from 25(OH)D measurements were extracted from Fleury Laboratory Database (the largest private laboratory in Brazil). All data were de-identified before analysis. A total of 59,812 samples (45,600 females and 14,212 males) collected from adults between January 2010 and July 2011 was analyzed. The mean age was 50.47 [plusmn] 14.0 years (18-70). 25(OH)D concentrations were determined by LC-MS/MS. [bold]Results:[/bold] Fifty nine percent of 25(0H)D values were below 30 ng/mL. The mean levels of 25(OH)D were lower after winter (25.9 ng/mL) and statistically different from that observed after summer (35.5 ng/mL; [italic]p[/italic][lt]0.001), a 27% reduction. A significant decrease in the mean 25(OH)D was observed with age in the female group, but not in the male group. [bold]Conclusion:[/bold] Although Brazil is mainly a tropical country, high prevalence of vitamin D insufficiency was observed in the city of Sao Paulo.[br][br](1) Holick MF et al., J Clin Endocrin Metab 2011; 96(7):1911. (2) Maeda SS et al., BMC Endocrine Disorders 2010; 10:12. (3) Maeda SS et al., Braz J Med Biol Res 2007;40:1653.[br][br]Nothing to Disclose: RPMB, CB, OHN, MT, JdS, VMC, JGHV 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 868 306 2244 MON-353 PO07-01 Monday 1913 2012


1909 ENDO12L_MON-354 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Vitamin D and Prostate Cancer Subhashini Yaturu, Sonya Zdunek, Barbara Youngberg Stratton VA Medical Center, Albany, NY; Stratton VA Medical Center, Albany, NY; Stratton VA Medical Center, Albany, NY Prostate cancer is the second common cancer in men worldwide and is the 2[sup]nd[/sup] leading cause of cancer deaths in men in the United States. Vitamin D is very important for normal physiology and is considered to have anticancer properties. Anticancer activities of vitamin D have been suggested to act mainly through its nuclear receptor or vitamin D receptor. In this retrospective study we compared the vitamin D levels in subjects with Prostate cancer with that of age matched men without prostate cancer. Study subjects include 480 in each group with a mean age of 73 and a mean creatinine of 1.05 and 1.15. In contrast to the belief, we did not find a significant difference in 25, (OH) vitamin D as well as 1, 25, (OH) vitamin D levels. Levels of 25, (OH) vitamin D were 28.4 [plusmn] 0.54 in subjects with prostate cancer and 28.05 [plusmn] 0.62 in subjects without prostate cancer. Levels of 1, 25, (OH) vitamin D were 47.2 [plusmn] 6.8 in subjects with prostate cancer and 47.1 [plusmn] 7.11 in subjects without prostate cancer. Vitamin D levels correlate with age ([italic]r[/italic] = 0.11 [italic]P[/italic] =0.01) and negative association with BMI ([italic]r[/italic] = 0.11 [italic]P[/italic] =0.01), glucose and presence of diabetes[italic] r[/italic] = 0.11 [italic]P[/italic] =0.001). We conclude that low vitamin D levels may not have a causal effect but simple association with age.[br][br]Sources of Research Support: Salary support by Stratton VA Medical Center.[br][br]Nothing to Disclose: SY, SZ, BY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1556 306 2245 MON-354 PO07-01 Monday 1914 2012


1910 ENDO12L_MON-355 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) ROC Curve Analysis for Evaluation of Cutoff Levels of Vitamin D Deficiency with Different Methodologies Ana M Sequera, Gabriela F Ruibal, Eleonora Nunez Chavarria, Hilda Farelo, Anabella Odriozola, Maria P Fosatti, Delia Pelletier, Paula Esteban, Silvia Corti, Maria Aguet, Cecilia Aguirre, Fernanda Aseguinolaza, Gustavo Benitez, Sofia Coniglio, Erich Fradinger, Romina Fiorenzano, Ana Fontanazza, Maria E Gonzalez, Nanci Gutierrez, Carolina Justo, Maria Lescurat, Marcela Miro, Silvana Montero, Hugo Molinas, Clelia Paredes, Adriana Pittau, Amelia Reverendo, Cintia Tarifa, Samuel Barcudi, Ana Canteli, Cristina Calderon, Gisella Di Prieto Reveand, Angel Galassi, Valeria Garcia, Julieta Gimenez, Laura Guerrero, Nadia Kogovsek, Gabriela Munoz, Marcela Niro, Gabriel Ramos, Mariana Rocca, Natalia Rossi, Cecilia Fenili, Mirta Gurkinkiel, Andrea Kozak Argentine Society for Endocrinology and Metabolism, Buenos Aires, Argentina Hypovitaminosis D, both in young adults and elderly and mainly in big cities, has an important incidence in the general population. The increase in number of patients evaluated for assessing Vitamin D deficiency have led to the development of automatic systems, faster and less laborious. This leads to the verification of the application of the consensus cutoff levels agreed for levels of 25-OH Vitamin D (VIT D) in these new methodologies. Objective: To compare and establish by ROC Curve analysis the consensus cutoff values for deficiency of VIT D accepted for RIA DIASORIN and the cutoff limits with greater sensitivity(S) and specificity(Sp) in the other systems evaluated.[br]Materials and Methods: 254 serum samples from healthy adults (18 - 45 years) without VIT D intake were included. Subjects with severe organic insufficiencies, or those on medication for bone metabolism alterations or VIT D were excluded.[br]Samples were aliquoted and stored at -20[ordm]C during the study. VIT D was tested by three methods measuring total VIT D: radioimmunoassay (RIA, DIASORIN); chemiluminiscence (CLIA, ADVIA Centaur, Siemens Diagnostics), and electrochemiluminiscence (ECLIA, Cobas e411, Roche Diagnostics). Samples were collected from three geographical regions: North (n:43) 26[deg]-27[deg] South(S) latitude, Centre (n:102), 31[deg]-36[deg] S latitud and South (n:109) 38[deg]-51[deg] S latitude. Cutoff levels with best S and Sp were calculated by ROC curve analysis. Samples with VIT D values [le] 20.0 ng/ml by RIA were interpreted as true positives.[br]Results: VIT D values (median and range) by RIA were 23.0 (6.2-94.0) ng/ml; by CLIA were 19.7 (4.9-49.8) ng/ml and by ECLIA were 22.5 (3.0-60.4) ng/ml. Significant differences were found between RIA and CLIA and between CLIA y ECLIA (p[lt] 0.05, Friedman, Dunn test).[br]ROC Curve analysis showed that samples with VIT D [le] 20.0 ng/ml by RIA would have by CLIA a cutoff of 19.62 ng/ml (S: 83.3 %, Sp: 56.5, AUC: 0.727) and by EQLIA a cutoff of 19.60 ng/ml (S: 70.8 %, Sp: 81.7, AUC: 0.818).[br]Conclusions: Our results showed that a cutoff of 19.6 ng/ml for CLIA and ECLIA would be equivalent to the value of 20 ng/ml used for define VIT D deficiency. Although the election of automatic systems against manual RIA is advisable for definition of VIT D deficiency, we consider that more studies are required for establish their utility in the follow up of patients under supplementation treatment.[br][br]Nothing to Disclose: AMS, GFR, ENC, HF, AO, MPF, DP, PE, SC, MA, CA, FA, GB, SC, EF , RF, AF, MEG, NG, CJ, ML, MM, SM, HM, CP, AP, AR, CT, SB, AC , CC, GDPR, AG, VG, JG, LG, NK, GM, MN, GR, MR, NR, CF, MG, AK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1097 306 2246 MON-355 PO07-01 Monday 1915 2012


1911 ENDO12L_MON-356 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Young Adult Male Body Composition at a Presence of Vitamin D Insufficiency Ieva Gailyte, Lina Lasaite, Gintautas Kazanavicius, Romualdas Tomas Preiksa Lithuanian University of Health Sciences, Kaunas, Lithuania; Lithuanian University of Health Sciences, Kaunas, Lithuania Population studies show that vitamin D level is related to body mass index, waist circumference and it is negatively related to parathyroid hormone (PTH). But it[apos]s relation to body composition is still controversial and not completely clear. The objective of the study was to measure the severity of vitamin D deficiency and PTH concentration in young healthy men in the period of winter and to establish the relationship with body composition in these men.[br]Methods and patients. Young healthy men from different regions of Lithuania, who were serving as military conscripts for one year were recruited for the study. 123 of them (age 18-27 years) agreed to participate. Fasting overnight blood samples were obtained in winter period. The 25(OH)D3 and PTH levels were measured by an electrochemoliuminescent immunological assay [quot]ECLIA[quot] ([bdquo]Roche Elecsys 1010/2010[ldquo]). Measuring of body composition was done by Jawon Medical BODYPASS X-SCAN BIA analyzer.[br]Results. More than a half of young healthy men in winter period had vitamin D insufficiency (12-30 ng/ml), 43% had deficiency ([lt]12 ng/ml), and only 1.7% had sufficient recommended concentration ([gt]30 ng/ml).[br]PTH concentration was significantly higher in those persons who have vitamin D deficiency than in those who have higher, but still not sufficient vitamin D concentration (2.6[plusmn]1.2 vs. 2.9[plusmn]1.0 pg/ml; p=0.02).[br]In persons with higher vitamin D concentration weight (73.6[plusmn]7.4 vs. 77.9[plusmn]8.8 kg; p=0.01), body mass index (22.6[plusmn]1.8 vs. 23.5[plusmn]2.3 kg/m[sup2]; p=0.048), lean body mass (61.6[plusmn]5.5 vs. 64.4[plusmn]6.8 kg; p=0.02) and body water (44.3[plusmn]3.9 vs. 46.3[plusmn]4.9 kg; p=0.021) were significantly higher than in persons with lower vitamin D concentration. But there was no significant difference in fat body mass between persons who have higher and lower vitamin D concentration.[br]Significant correlations were detected between vitamin D concentration and weight (r=0.244; p=0.009), body mass index (r=0.222; p=0.018), lean body mass (r=0.240; p=0.010), body water (r=0.240; p=0.010).[br]Conclusion. In this study almost al healthy young men in winter period had vitamin D deficiency or insufficiency and only 1.7% of them had sufficient recommended vitamin D concentration.[br]Vitamin D was negatively correlated with PTH.[br]Relations between Vitamin D concentration and body mass index and body composition in healthy not-obese young persons could be different from obese and overweight persons.[br][br]Nothing to Disclose: IG, LL, GK, RTP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1036 306 2247 MON-356 PO07-01 Monday 1916 2012


1912 ENDO12L_MON-357 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Vitamin D3 and Androgen Status in Men Juraj Payer, Adriana Banarova, Peter Jackuliak, Tomas Koller University Hospital and Medical Faculty of Comenius University, Bratislava, Slovakia (Slovak Republic) Objective: 25-OH vitamin D3 (D3) possibly affects testosterone production in Leydig cells. Although pathogenesis of this relationship is not well understood, in some cross sectional studies the level of D3 correlates positively with total testosterone (TT) and free androgen index (FAI). In the European Male Ageing Study positive association of D3 with secondary and compensated hypogonadism was confirmed. In other study supplementation of D3 led to increase of testosterone levels in men.[br]Aim of the study: was to determine the relationship between D3 levels and levels of TT, luteninizing hormone (LH), follicle-stimulating hormone (FSH) and FAI in men with acute coronary syndrome. Secondary aim was to divide measured values into quartiles of physiological range.[br]Methods: In 38 men with average age 72 years admitted to 5th Department of Internal Medicine with acute coronary syndrome we determined levels of D3 by chromatograhy, TT, LH and FSH by electrochemiluminescence and calculated FAI (TT/SHBG x100). Patients with severe liver disease and malignancy were excluded. Patients did not receive antiandrogen therapy and vitamin D3 supplements. The correlation statistic was done by MedCalc system.[br]Results: The average level of TT was 12.93 nmol/l (interval 3.22-24.15 nmol/l), FAI 32.33% (14- 47.26%), FSH 9.03 IU/l (2.12-45.14), LH 7.92 IU/l (2.26-34.26) and D3 21 ug/l (4 [ndash] 52.7 ug/l). We did not confirmed correlation of D3 with neither TT (r: -0.03; p: 0.86), nor FAI (r: -0.13; p: 0.43) by statistical methods. We also did not confirm correlation of D3 with FSH or LH. By dividing measured values into quartiles of physiological range we figured out that 100% of patients had levels of D3 vitamin in 2 lowest quartiles, 84% of patients had TT in 2 lowest quartiles and 97% of patients had FAI in 2 lowest quartiles of physiological range.[br]Conclusion: In our study we did not confirme correlation of D3 with TT, FAI, FSH and LH by statistical methods. Limitations of our study were small and specific sample (patients with acute coronary syndrome) and high average age. By dividing patients into quartiles of physiological range we can conclude that most patients with low levels of TT and FAI had also low levels of D3. More investigation is necessary to explain the relationship between D3 and androgen status in men.[br][br]Nothing to Disclose: JP, AB, PJ, TK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1262 306 2248 MON-357 PO07-01 Monday 1917 2012


1913 ENDO12L_MON-358 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Association between Vitamin D Status and Physical Function in the Severely Obese Aftab Khattak, Tomas Ahern, Emer O[apos]Malley, Dunlevy Colin, Kilbane Mark, Donal O[apos]Shea, McKenna Malachi St Columcille[apos]s Hospital, Loughlinstown, Ireland; St Vincent[apos]s University Hospital, Elm Park, Ireland; St Vincent[apos]s University Hospital, Elm Park, Ireland; St Vincent[apos]s University Hospital, Elm Park, Ireland [italic]Introduction[/italic][br]Severe obesity (BMI[gt]40kg/m[sup]2[/sup]) has increased five-fold in prevalence over the past 15 years in the USA. Vitamin D deficiency is associated with physical dysfunction in the elderly. Obese people commonly have low serum 25hydroxyvitamin D (25OHD) due to sequestration in adipose tissue or reverse causation. We tested the hypothesis that poor vitamin D status is associated with physical inactivity and physical dysfunction in severely obese people.[br][italic]Methods[/italic][br]We determined 25OHD concentrations, recorded the hours per week spent exercising and measured the times taken to complete tests of physical function in 254 severely obese people. The tests consisted of ascending and descending a step 50 times and walking 500 m. Based on the recent Institute of Medicine report we defined vitamin D status as follows: at risk of deficiency (25OHD [lt]30 nmol/L); within range of requirement (25OHD 30-50 nmol/L); and above sufficiency limit (25OHD [ge]50 nmol/L). Mann-Whitney U, Kruskall-Wallis and Spearman analyses were performed. Data are presented as means[plusmn]SD and correlation coefficients (r).[br][italic]Results[/italic][br]The average 25OHD and BMI for the entire cohort were 35.9[plusmn]15.1 nmol/L and 51.1[plusmn]8.4 kg/m[sup]2[/sup] respectively. 72.5% completed the 500m walk test and 81.0% the 50 step test. Serum 25OHD was higher in 50 step test completers (36.8[plusmn]13.9 nmol/L) than in non-completers (29.7[plusmn]13.0 nmol/L, p=0.003).[br]Forty percent were at risk of being deficient, 40% were within range of requirement and 20% were above the sufficiency limit. Weekly exercise time increased with improving vitamin D status: 1.4[plusmn]2.5, 1.4[plusmn]2.0 and 3.1[plusmn]3.4 hours respectively (p=0.008). In test completers 500 m walk time decreased with improving vitamin D status: 7.0[plusmn]1.4, 6.8[plusmn]1.1 and 6.3[plusmn]1.1 minutes respectively (p[lt]0.001). Similarly 50 step time decreased with improving vitamin D status: 124[plusmn]29, 113[plusmn]25 and 114[plusmn]23 seconds respectively (p=0.039). Serum 25OHD correlated significantly with exercise time (r=0.17, p=0.02), with 500 m walk time (r=-0.37, p[lt]0.001) and with 50 step time (r=-0.16, p=0.04).[br][italic]Conclusions[/italic][br]In this cohort of severely obese people the prevalence of hypovitaminosis D was 40%. Vitamin D status had a strong relationship with physical inactivity and physical dysfunction in the severely obese. We conclude that vitamin D deficiency may contribute toward the risk of obesity related complications and suggest that studies exploring the effects of vitamin D supplementation in this cohort are warranted.[br][br]Sources of Research Support: The Irish Heart Foundation and the Irish Health Research Board in the form of unrestricted research grants.[br][br]Nothing to Disclose: AK, TA, EO, DC, KM, DO, MM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1264 306 2249 MON-358 PO07-01 Monday 1918 2012


1914 ENDO12L_MON-359 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Relation of Vitamin-D Status with Insulin Resistance and Systemic Inflammation in Individuals with Normal Glucose Tolerance, Prediabetes and Diabetes: An Eastern India Perspective Deep Dutta, Anirban Sinha, Indira Maisnam, Ankit Shrivastava, Sujoy Ghosh, Satinath Mukhopadhyay, Subhankar Chowdhury Institute of Post-Graduate Medical Education and Research, Calcutta, India Introduction: Vitamin-D (Vit-D) deficiency may have an important role in the pathogenesis of diabetes by acting through promoting lipogenesis, enhancing [beta]-cell loss due to increased systemic inflammation and through increased circulating levels of PTH. This study is an initial part of the study to evaluate the role of correcting Vit-D deficiency in prevention of progression of prediabetes to diabetes (CTRI/2011/091/000192).[br]Methods: 200 individuals of prediabetes (PD) [gt]30 years age, not on medication with impaired fasting glucose (IFG) and/or impaired glucose tolerance (IGT) on 2 occasions a week apart, were selected and compared to 100 age and BMI matched patients of diabetes (DM) and 50 normal individuals (NI) by population screening. Serum vit-D was estimated using chemiluminescent microparticle immunoassay (CLIA) (Architect i1000, Abbott,USA). Serum fasting insulin and hs-cRP was estimated using CLIA (Immulite-1000,USA).[br]Results: 76%(76/100), 72%(144/200) and 62%(31/50) of individual of DM, PD and normal glucose tolerance (NGT/NI) had vitamin-D deficiency/insufficiency. There was significant difference in waist hip ratio (0.84[plusmn]0.07, 0.9[plusmn]0.07, 0.93[plusmn]0.04; P=0.018) and waist height ratio (0.54[plusmn]0.05, 0.57[plusmn]0.07, 1.36[plusmn]0.32; P[lt]0.001) among NI, PD and DM groups. Serum 25(OH)vit-D level was not significantly different among the 3 groups (P=0.1). hs-cRP was significantly different among NI, PD and DM groups (1.28[plusmn]1.7, 4[plusmn]3.1 and 7.46[plusmn]3.6mg/L; P[lt]0.001). In individuals with PD, there was significant difference in insulin resistance (HOMA2-IR: P=0.04; QUICKI Insulin Sensitivity Index: P=0.03) but not in estimated pancreatic beta cell function (HOMA2[beta]; P=0.21) and hs-cRP (P=0.7) among those with 25(OH)vit-D[lt]10,11-20,21-30 and [gt]30ng/ml. After adjusting for BMI, there was an inverse correlation between vit-D and HOMA2-IR (r=-0.26; P=0.01), and positive correlation with QUICKI (r=0.29; P=0.007) among individuals with PD but not in NI and DM groups. There was inverse correlation between vit-D and hs-cRP among individuals with DM (r=-0.47;P=0.07) but not in NI and PD group.[br]Conclusion: Vit-D deficiency was more common in individuals with prediabetes and diabetes as compared to normal individuals. Vit-D deficiency predicts insulin resistance among individuals with prediabetes but not in normal individuals or patients with diabetes. Inverse correlation between systemic inflammation (hs-cRP) and vit-D was seen only in patients with diabetes.[br][br]Nothing to Disclose: DD, AS, IM, AS, SG, SM, SC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 734 306 2250 MON-359 PO07-01 Monday 1919 2012


1915 ENDO12L_MON-360 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Modulation of [italic]CYP2R1 [/italic]Expression: A Potential Mechanism for Determining Vitamin D Requirements in Aging and Obesity Jeffrey D Roizen, Isaac Sasson, Michael A Levine The Children[apos]s Hospital of Philadelphia, Philadelphia, PA; University of Pennsylvania Perelman School of Medicine, Philadelphia, PA Vitamin D insufficiency is a worldwide public health problem that leads to rickets in children and increased risk of fracture in the aging population. Vitamin D status is assessed by measuring circulating levels of 25(OH)-vitamin D (25OHD) a metabolite reflecting the first step in activation of vitamin D obtained either through the diet or produced in the skin in response to UV light. Although low levels of 25OHD generally indicate inadequate supply of vitamin D, we hypothesized that circulating 25OHD might also be influenced by altered expression of [italic]CYP2R1[/italic], which encodes the principle hepatic vitamin D 25-hydroxylase. To test this hypothesis, we assessed expression of [italic]Cyp2r1[/italic] mRNA in livers of obese and aging mice, two models of vitamin D insufficiency. We used a mouse model of diet-induced obesity in which mice were fed a high-fat diet (30% of intake) ad libitum from 3 weeks of age to replicate the effects of the [ldquo]western[rdquo] diet on human physiology. Control mice were fed a standard-fat diet (10% fat). Both diets contained normal amounts of vitamin D. Significant differences in weight were observed after 10 weeks of diet (lean vs obese groups: 20.4 [plusmn] 1.9 vs 30.8 [plusmn] 0.7 gms at 13 weeks; 23.4 [plusmn] 2.3 vs 34.6 [plusmn] 4.4 gms at 15 weeks; 23.6 [plusmn] 2.1 vs 34.3 [plusmn] 4.3 gms at 21 weeks; and 22.7 [plusmn] 0.5 vs 48.8 [plusmn] 3.6 gms at 24 weeks (mean [plusmn] SD). Two-way ANOVA was significant for effects of age and diet (p[lt]0.001), and post-test Bonferroni showed significant (p [lt] 0.001) differences between lean and obese weights at all ages. We used RT-qPCR to measure the relative abundance of hepatic [italic]Cyp2r1[/italic] mRNA normalized to [beta] actin mRNA. There was a positive trend for increasing relative abundance of [italic]Cyp2r1[/italic] mRNA for lean mice from 13 to 24 weeks (1 [plusmn] 0.16, 1.09 [plusmn] 0.21, 1.26[plusmn]0.25, 1.62[plusmn]0.38) and no change in relative abundance of [italic]Cyp2r1[/italic] mRNA in obese mice (0.87[plusmn]0.35, 0.91[plusmn]0.17, 0.93[plusmn]0.26, 0.91[plusmn]0.17, mean [plusmn] SD). There were significant differences between the lean and obese mice by two-way ANOVA (p [lt] 0.01), with age effects not reaching significance (p=0.15). There was a decrease in the relative abundance of [italic]Cyp2r1[/italic] mRNA in obese mice relative to lean mice that reached significance (p [lt] 0.05, Bonferroni post-test analysis) by 24 weeks of age. These results suggest that hepatic expression of the [italic]Cyr2r1[/italic] gene increases with aging in lean mice, but not in obese mice. We propose that decreased expression of hepatic [italic]Cyp2r1[/italic] may account in part for lower circulating levels of 25OHD in obesity.[br][br]Nothing to Disclose: JDR, IS, MAL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2111 306 2251 MON-360 PO07-01 Monday 1920 2012


1916 ENDO12L_MON-361 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Effect of Fat on Serum 25-Hydroxyvitamin D Levels after a Single Oral Dose of Vitamin D: A Double-Blind Randomized Placebo-Controlled Study Fabiana Viegas Raimundo, Maria Augusta Lang, Luciano Scopel, Natalia Marcondes, Gustavo Adolpho Moreira Faulhaber, Tania Weber Furlanetto HCPA, Porto Alegre, Brazil; UFRGS, Porto Alegre, Brazil Introduction: Vitamin D supplements are useful to prevent and treat vitamin D deficiency. As this molecule is hydrophobic its oral absorption could vary according to the fat content of the meal it is consumed with, although some specialists believe there is no evidence to recommend its ingestion with food.[br]Objective: To evaluate serum 25-hydroxyvitamin D [25(OH)D]levels after the oral intake of a single dose of cholecalciferol, with three different content of fat meals, or placebo.[br]Subjects and Methods: A double-blind placebo controlled trial included sixty-four healthy medical residents of a university hospital in Porto Alegre, latitude 30[sup]0[/sup], Brazil, divided in four groups. Three groups received 50,000 IU of cholecalciferol with 490 to 550calories meals containing different lipid contents: Group 1 (G1): 0g, Group 2 (G2): 15g, Group 3(G3): 30g, and one group (G4) received placebo, according to randomization. Serum 25-hydroxyvitamin D [25(OH)D], parathyroid hormone (PTH), serum total calcium, albumin, magnesium and creatinine levels; urinary calcium, magnesium and creatinine levels were measured at baseline, and after 14 days.[br]Results: Mean serum 25(OH)D levels were low in all groups (G1= 17.84[plusmn]6.78ng/mL; G2=14.03[plusmn]5.68ng/mL; G3= 15.16[plusmn]6.03ng/mL; G4=14.31[plusmn] 5.39 ng/mL). After 2-weeks, vitamin D given during breakfast increased mean variation of serum 25(OH)D levels, when compared to placebo: G1(5.28 [plusmn] 3.24 ng/mL)x G4(1.14[plusmn] 2.00ng/mL), p[lt]0.001; G2(8.59 [plusmn] 2.71ng/mL)x G4, p[lt]0.000; G3(8.64[plusmn] 2.98ng/mL)x G4, p[lt]0.000. Nevertheless, the intake of fat during vitamin D administration increased further the variation of mean serum 25(OH)D levels: G1XG2, p[lt]0.007; and G1XG3, p[lt]0.008. Serum PTH and 25(OH)D levels were inversely correlated (r=-0.237; p=0.008).[br]Conclusion: After two weeks, a single oral dose of vitamin D was effective in increasing mean serum 25(OH)D levels when given with food, especially if it contained fat. These findings can have important implications to define the adequate dietary intake of vitamin D.[br][br]Sources of Research Support: Funding: Fundo de Incentivo [agrave] Pesquisa e Eventos, Hospital de Cl[iacute]nicas de Porto Alegre, Porto Alegre, RS, Brazil; (NCT01519986).[br][br]Nothing to Disclose: FVR, MAL, LS, NM, GAMF, TWF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1058 306 2252 MON-361 PO07-01 Monday 1921 2012


1917 ENDO12L_MON-362 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Lipodystrophy Does Not Seem To Affect Vitamin D Plasma Levels in HIV-Infected Patients Treated with Combined Antiretroviral Therapy Paula Freitas, Eva Lau, Ana Cristina Santos, Maria Joao Matos, Flora Correia, Jorge Pereira, Esteban Martinez, Antonio Sarmento, Jose Luis Medina, Davide Carvalho Centro Hospitalar S[atilde]o Jo[atilde]o, Faculty of Medicine University of Porto, Porto, Portugal; Faculty of Medicine of Porto, Porto, Portugal; Centro Hospitalar S[atilde]o Jo[atilde]o, Porto, Portugal; Hospital Clinic, University of Barcelona Medical School, Barcelona, Spain; Centro Hospitalar S[atilde]o Jo[atilde]o, Faculty of Medicine University of Porto, Porto, Portugal [bold]Introduction:[/bold] Subcutaneous tissue is responsible for 90% of vitamin D synthesis. Its deficiency is prevalent in HIV-infected patients and combined antiretroviral therapy (cART) by its association with lipodystrophy may contribute to this situation. [bold]Aims[/bold]: To compare vitamin D plasma levels in HIV infected patients with or without lipodystrophy (clinically and FMR defined) and according to body composition distribution (no lipodystrophy; isolated central fat accumulation; peripheral lipoatrophy; mixed forms of lipodystrophy). [bold]Methods:[/bold] Cross-sectional study. DXA body composition, metabolic parameters and 25-OH-vitamin D were evaluated.in 133 HIV1-infected patients on cART.[bold] Results:[/bold] No differences in vitamin D median according to the presence of lipodystrophy, body composition, gender, NRTI (nucleoside reverse transcriptase inhibitor) or NNRTI (non-nucleoside reverse transcriptase inhibitor) treatment, smoking status, CDC class or viral load were observed. Patients under PI (protease inhibitor) had higher levels of vitamin D [20.0 ng/mL (13.0-28.0) vs 15.0 ng/mL (7.0-23.0), p=0.027]. When we evaluated vitamin D according to 3 different plasma levels groups ([lt]20; 21-30; [gt]30 ng/mL), also no differences were found according to the presence of lipodystrophy, body composition, gender, age, HIV infection duration, PI, NNRTI or NRTI treatment, fat mass (total, upper limbs, lower limbs, trunk), non-fat mass or total bone mineral density (TBMD). Patients with longer duration antiretroviral therapy had higher levels of vitamin D [1) 7.0 (4.0 to 9.0), 2) 5.0 (2.0 to 10.0), 3) 10.0 (7.0 to 11.0), p = 0.020]. No significant associations were found between vitamin D and age, HIV infection or ART duration, CD4, TNF-a, total cholesterol, HDL-C, LDL-C, non-HDL-C, triglycerides, apo B, apo A1, Lp(a), A1c, insulin, HOMA, QUICKI, leptin, adiponectin, fat mass (total, upper limbs, lower limbs, trunk), non-fat mass or TBMD. [bold]Conclusion:[/bold] Lipodystrophy does not seem to influence vitamin D levels, although patients with longer duration of antiretroviral therapy had higher levels.[br][br]Nothing to Disclose: PF, EL, ACS, MJM, FC, JP, EM, AS, JLM, DC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 115 306 2253 MON-362 PO07-01 Monday 1922 2012


1918 ENDO12L_MON-363 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Vitamin D Supplementation and Global Proteomic Profiling: A Randomized, Double-Blind, and Placebo-Controlled Clinical Trial in Overweight/Obese Young African Americans Haidong Zhu, Jim Mobley, Dongquan Chen, Dehuang Guo, Samip J Parikh, Wendy Bollag, Yanbin Dong Georgia Health Sciences University, Augusta, GA; University of Alabama at Birmingham, Birmingham, AL; University of Alabama at Birmingham, Birmingham, AL; Georgia Health Sciences University, Augusta, GA; Georgia Health Sciences University, Augusta, GA Backgrounds:[br]Although vitamin D is considered to be involved in a broad range of bodily functions beyond bone health, the biological effects of vitamin D supplementation need to be defined. We sought to examine the impact of vitamin D supplementation on the serum proteome by means of mass spectrometry in overweight/obese African Americans having vitamin D insufficiency. We also aimed to identify new serum biomarkers of vitamin D supplementation using proteomic profiling.[br]Methods:[br]A randomized, double-blind, placebo-controlled 16-week clinical trial was conducted in 57 overweight/obese young African-American adults (Clinicaltrial.org ID: NCT01141192). Participants were randomly assigned to either the placebo or vitamin D (oral 60,000 IU/month equivalent to [sim]2,000 IU/day) group. Sera were analyzed by LC-ESI-MS/MS.[br]Results:[br]Following 16 weeks of vitamin D3 intervention (n = 22, 31[plusmn]10 years of age), serum 25(OH)D concentrations increased from 35.4[plusmn]11.3 at baseline to 103.7[plusmn]31.5 nmol/L at posttests (p[lt]0.0001). A total of 346 distinct serum proteins were identified per subject with [ge]99% confidence, and [ge] 2 peptides per protein. Statistical analyses revealed 22 proteins that were either increased or decreased in abundance with greater than 1.5 fold changes by vitamin D supplementation. Among them, the top ones included keratin type I cytoskeletal 10 (5.00 fold, p=0.005), keratin type II cytoskeletal 1 (4.82 fold, p=0.005), titin (2.45 fold, p=0.009), D4 zinc and double PHD fingers family 1, (2.43 fold, p=0.021), T-Dicer (2.33 fold, p=0.020), kinesin family member 22 (-2.22 fold, p=0.026), zinc finger protein 536 (2.10 fold, p=0.011), glutamine synthetase (2.00 fold, p=0.041), [alpha]-kinase 3 (1.83 fold, p=0.038), hyaluronan mediated motility receptor (1.73 fold, p=0.043), serine/threonine-protein kinase MRCK-[beta] (1.73 fold, p=0.058), WD repeat-containing protein 52 (-1.69 fold, p=0.035), [alpha]-1,6-mannosylglycoprotein 6-[beta]-N-acetylglucosaminyltransferase A (1.69 fold, p=0.039), and semaphorin-3 (1.52 fold, p=0.056). Placebo (n=23, 31 [plusmn] 2 years of age) did not show any effects on these proteins.[br]Conclusions:[br]Our data provide a possible basis for defining the broader biological effects of vitamin D supplementation. The present study also demonstrates that protein mass profiling may be an effective tool for the discovery of possible novel mechanisms and biomarkers of vitamin D repletion. Future replication studies and dose-responsive clinical trials are warranted.[br][br]Nothing to Disclose: HZ, JM, DC, DG, SJP, WB, YD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 981 306 2254 MON-363 PO07-01 Monday 1923 2012


1919 ENDO12L_MON-364 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Increased 24 Hydroxylation Does Not Explain Low Plasma Vitamin D Levels in Obese Adolescents Noelle S Larson, Candace Percival, Jill Emerick, David W Larson, Karen Vogt, Ronald Horst, Merrily Poth Walter Reed National Military Medical Center, Bethesda, MD; Uniformed Services University of the Health Sciences, Bethesda, MD; Heartland Assays, Ames, IA [bold]Background:[/bold] Obesity has been shown to be associated with low plasma levels of 25 hydroxy vitamin D in both adult and pediatric populations. The reason for this apparent vitamin D deficiency is not clear. We hypothesized that adipose, known to be a highly metabolically active tissue, might play an active role in the breakdown of vitamin D, contributing to low 25 hydroxy vitamin D levels observed in obese individuals. Since the primary pathway for vitamin D metabolism is through 24 hydroxylation, we used measurement of plasma 24,25 dihydroxy vitamin D levels as a potential marker of such a process.[br][bold]Methods:[/bold] We measured two vitamin D metabolites (25 hydroxy and 24,25 dihydroxy vitamin D) in the plasma of 23 obese and 9 normal weight adolescents, age 10-18 years. Subjects filled out questionnaires regarding typical intake of dairy and vitamin D containing supplements as well as typical sun exposure patterns. We gathered anthropometric measurements, blood pressure, HOMA and lipid data for all subjects.[br][bold]Results:[/bold] Our data confirmed that mean 25 hydroxy vitamin D levels are lower in obese individuals than healthy weight controls (p = 0.03). The plasma concentration of 25 hydroxy vitamin D correlated with BMI percentile (p=0.03). Dietary intake was not different between the two groups, but we found a strong correlation between 25 hydroxy vitamin D level and sun exposure score (p=0.007). We did find a statistically significant relationship between the ratio of 24,25 dihydroxy: 25 hydroxy vitamin D and BMI (p=0.03). However, contrary to our hypothesis, this ratio was lower in the obese group and 24,25 hydroxy D levels were positively correlation with 25 hydroxy vitamin D level (p=0.0009, r[sup]2[/sup] = 0.31) in all subjects.[br][bold]Conclusions:[/bold] We could not confirm our hypothesis that lower 25 hydroxy vitamin D levels in overweight adolescents are a result of increased 24 hydroxylation. However, we cannot rule out the possibility that adipose is able to catabolize vitamin D in a manner not reflected by plasma 24,25 dihydroxy vitamin D. Our data also suggest that the differences we observed in plasma vitamin D levels in our population may be at least partly due to differences in sun exposure.[br][br]Nothing to Disclose: NSL, CP, JE, DWL, KV, RH, MP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1885 306 2255 MON-364 PO07-01 Monday 1924 2012


1920 ENDO12L_MON-365 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) The Relationship between Vitamin D Status and Body Composition in Pre- or Early Pubertal Nonobese Children Young Ah Lee, Ji Young Kim, Min Jae Kang, Shin Mi Kim, Seung Joon Chung, Choong Ho Shin, Sei Won Yang Seoul National University Children[apos]s Hospital, Seoul, Republic of Korea; Seoul Red Cross Hospital, Seoul, Republic of Korea Background: The 2010 Dietary reference intakes for Koreans lowered adequate intake (AI) of vitamin D to 200 IU/day in childhood due to lack of evidence. We evaluated the prevalence of vitamin D deficiency and risk factors for low vitamin D status. We also investigated the relationship between vitamin D status and body composition and bone mineral density (BMD) in pre- or early pubertal nonobese children. We purposed to provide evidence for determining Korean AI of vitamin D.[br]Methods: A total 100 nonobese children (9.3 [plusmn] 1.9yrs, 45 boys, 71 prepuberty) living in Seoul, Korea were enrolled in wintertime (n = 38, December, 2010 to March, 2011) and summertime (n = 62, June to September, 2011). Z score of the bone mineral content (Z_BMC), fat mass (Z_FM), lean tissue mass (Z_LTM), and total body (Z_TB) and L-spine 1-4 (Z_L1-4) BMD using Dual-energy X-ray absorptiometry were calculated according to the reference for Korean children. Daily calcium (mg/day) and vitamin D intake (U/day) were evaluated by dietitian. The serum level of 25(OH)D below 20ng/mL was defined as vitamin D deficiency. Questionnaires were used to assess regular physical activity and outdoor activities (hrs/week).[br]Results: The prevalence of vitamin D deficiency was 47.4% in winter and 17.7% in summertime. In wintertime, vitamin D status negatively associated with vitamin D intake ([italic]P [/italic]= 0.019) and less outdoor activities ([italic]P[/italic] = 0.015), after adjusting for sex and puberty. About half of children with vitamin D intake below 800 IU/day were vitamin D deficient in winter. The level of 25(OH)D negatively correlated with Z_BMC ([italic]P[/italic] = 0.023), Z_TB ([italic]P[/italic] = 0.018), and Z_L1-4 ([italic]P [/italic]= 0.043), after controlling for sex, puberty, Z_FM, physical activity and the percentage of the adequate calcium intake. However, the Z_FM and Z_LTM had no significant relationship with the level of 25(OH)D.[br]Conclusions: About half of pre- and early pubertal nonobese children were vitamin D deficient in wintertime. Considering the effect of vitamin D status on bone health in childhood, the importance of adequate vitamin D intake and outdoor activities needs to be emphasized. Current recommendation for AI of vitamin D (200IU/day) in Korean children seems not to be enough to prevent vitamin D deficiency, especially in wintertime.[br][br]Nothing to Disclose: YAL, JYK, MJK, SMK, SJC, CHS, SWY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1328 306 2256 MON-365 PO07-01 Monday 1925 2012


1921 ENDO12L_MON-366 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Is There a Relationship between Vitamin D and Thyroid Function in Obese Minority Children? Megan Kathleen Long, Ping Zhou, Jessica May Gold, Leslie Lam, Morri Markowitz Albert Einstein College of Medicine, Bronx, NY; Children[apos]s Hospital at Montefiore, Bronx, NY; Children[apos]s Hospital at Montefiore, Bronx, NY [bold]Background: [/bold]Vitamin D deficiency is often found in obese adolescents. Elevated TSH levels with a euthyroid state has also been linked with obesity. However, studies examining the relationship between vitamin D levels and thyroid function tests in obese pediatric populations are lacking.[br][bold]Objective[/bold]: To evaluate the relationship between 25-hydroxy vitamin D (abbreviated 25(OH)D) levels and thyroid function tests in an obese pediatric population.[br][bold]Methods[/bold]: This is an IRB-approved, retrospective study. Data from 333 obese patients, between the ages of 6 and 21 and of Hispanic or African American descent were included in the study. These subjects were referred to our endocrine clinic for evaluation of obesity, but were otherwise generally healthy. Data for 25(OH)D (mean 16.091[plusmn]9.78, range 5.33-90.80ng/mL), TSH (mean 1.92[plusmn]1.10, range 0.04-8.92 uU/mL), free T4 (1.28[plusmn]0.24, range 0.70-2.53 ng/dL), and anthropometric measures were abstracted. BMI and BMI z-score were calculated. Statistical analyses performed included determination of Pearson correlations and comparisons of means stratified by TSH level. For the latter, subjects were divided into low TSH (n=47, TSH [lt]1.0) and high TSH (n=16, TSH [gt]4.0) groups.[br][bold]Results[/bold]: 25(OH)D levels were negatively correlated with age (r= -0.21, p[lt]0.0001), height (r=-0.26, p[lt]0.0001), BMI (r=-0.21, p[lt]0.0001), and BMI z-score (r=-0.14, p=0.0071). There were no significant correlations between 25(OH)D and thyroid function tests. TSH was positively correlated with BMI z-score (r=0.152, p=0.0089). Differences in mean 25(OH)D levels between the high and low TSH groups were not statistically significant. However, BMI z-scores of the two groups were statistically different, with a mean BMI z-score of 2.17 for the low TSH group and 2.41 for the high TSH group (p=0.0466).[br][bold]Conclusions[/bold]: Our results support the previous findings that both vitamin D deficiency and TSH levels are associated with the degree of obesity. However, a direct association between thyroid function tests and 25(OH)D levels was not found in our obese children.[br][br]Garanty-Bogacka, B, Syrenicz, M, et al. Serum 25-hydroxyvitamin D (25-OH-D) in obese adolescents. Endokrynol Pol. 2011;62(6):506-11. Nader et al. Relationships Between Thyroid Function and Lipid Status or Insulin Resistance in a Pediatric Population. Thyroid, 2010; 20(12):1334-1339. Roos, Annemieke, Bakker, Stephan J. L, et al. Thyroid Function Is Associated with Components of the Metabolic Syndrome in Euthyroid Subjects. J Clin Endocrinol Metab, February 2007, 92(2):491[ndash] 496. Roth, CL, Elfers, C, et al. Vitamin d ddeficiency in obese children and its relationship to insulin resistance and adipokines. J Obes. 2011;2011:495101. Epub 2011 Dec 29.[br][br]Nothing to Disclose: MKL, PZ, JMG, LL, MM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1022 306 2257 MON-366 PO07-01 Monday 1926 2012


1922 ENDO12L_MON-367 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Association between Vitamin D Levels and Disease Severity in Patients with Obstructive Sleep Apnea Syndrome Yavuz Yalcin, Dilek Berker, Oya Topaloglu, Turkan Mete, Bulent Ciftci, Selma Firat Guven, Ayse Arduc, Bercem Aycicek Dogan, Serdar Guler Numune Education and Research Hospital, Ankara, Turkey; Atat[uuml]rk Chest Diseases and Thoracic Surgery Education and Research Hospital, Ankara, Turkey [bold]Objectives: [/bold]Obstructive sleep apnea syndrome (OSAS) is a widespread disorder characterized by recurrent, partial or complete episodes of apnea due to upper airway tract obstruction during sleeping period. Although there is some data about vitamin D levels in sleep disorders the issue has not been studied well in OSAS. Thus, we aimed to investigate the association between OSAS, disease severity and vitamin D levels in this study.[br][bold]Materials:[/bold] One-hundred fifty OSAS patients (50 with mild, 50 with moderate, 50 with severe) and 32 non-OSAS controls who visited Ankara Atat[uuml]rk Chest Disease Hospital for polysomnography from September 2010 to November 2011 were recruited to the study. Serum calcium, phosphorus, parathyroid hormone (PTH), and 25-hydroxy- Vitamin D levels were evaluated.[br][bold]Results: [/bold]Age, gender and body mass index were similar in both groups[bold]. [/bold]No differences were found between OSAS patients and controls with regard to calcium, phosphorus and PTH levels. There was also no difference between the two groups in terms of Vit D levels. But, the patients with severe OSAS had significantly lower levels of vitamin D as compared with other groups (p[lt]0.01). Also, number of patients with vitamin D levels less than 10 [micro]d/dL were found higher in OSAS patients than the controls (p[lt]0.02). We demonstrated that cut-off value for Apnea-hypopnea index (AHI) was 19.3 in patients with vitamin D levels less than 10 [micro]d/dL (55.8% sensitivity and 71.8% spesificity).[br][bold]Conclusion:[/bold] Our study demonstrated that Vitamin D deficiency may be frequent in patients with OSAS. Further studies are required to evaluate the effect of vitamin D supplementation on AHI in patients with vitamin D deficiency and OSAS.[br][br]Nothing to Disclose: YY, DB, OT, TM, BC, SFG, AA, BAD, SG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1580 306 2258 MON-367 PO07-01 Monday 1927 2012


1923 ENDO12L_MON-368 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Serum Sclerostin Levels Following Vitamin D and Calcium Supplementation Lisa Ceglia, Bess Dawson-Hughes, Anastassios G Pittas Tufts Medical Center, Boston, MA; Tufts University, Boston, MA [bold]Background[/bold]: Sclerostin is an osteocyte-derived protein that inhibits bone formation via inhibition of the Wnt pathway. Recent data suggest that circulating levels of sclerostin are associated with impaired bone remodeling and metabolic bone disease. Whether vitamin D and calcium supplementation may affect serum sclerostin levels has not been studied.[br][bold]Objective:[/bold] To examine the effect of 16-weeks of vitamin D[sub]3[/sub] supplementation with or without calcium on serum sclerostin concentration in adults at high risk for diabetes and early-diabetes, a population that has been reported to exhibit high levels of sclerostin.[br][bold]Methods[/bold]: In a double-masked, placebo-controlled trial, 92 adults with pre-diabetes were randomized in a 2x2 factorial design, to vitamin D[sub]3[/sub] 2000 IU daily or calcium carbonate 400 mg twice daily for 16 weeks. Serum sclerostin level was measured by ELISA.[br][bold]Results[/bold]: Participants had a mean age of 57 years, 51% were women, and 78% were white. Mean body mass index was 32 kg/m2, hemoglobin A1c was 5.9%, and baseline 25-hydroxyvitamin D level (25OHD) was 24.5 ng/mL. Mean change in 25OHD was 6.2 [plusmn] 1.0 ng/mL in the vitamin D group vs. -6.3 [plusmn] 1.0 ng/mL in the no-vitamin D group. There was no significant vitamin D [times] calcium interaction on change in sclerostin (p=0.16). After adjusting for several clinical factors including age, BMI, race and baseline sclerostin level, the mean change in sclerostin (pmol/L) did not differ between vitamin D vs. no vitamin D group (adjusted mean 0.89 [plusmn] 1.2 [n=46] vs. 0.09 [plusmn] 1.2 [n=46] respectively; p=0.58) There was also no significant difference in adjusted mean sclerostin level with calcium vs. no calcium supplementation (0.35 [plusmn] 1.2 [n=45] vs. 0.58 [plusmn] 1.2 [n=47] respectively; p=0.87).[br][bold]Conclusion[/bold]: In adults at high risk for diabetes, vitamin D and calcium supplementation did not have an effect on serum sclerostin level. Larger studies are needed to verify these results.[br][br]Sources of Research Support: The study was supported by R01DK76092 (to AGP), UL1 RR025752 (to Tufts University) and USDA Cooperative Agreement no. 58-1950-4-401 (to BDH).[br][br]Nothing to Disclose: LC, BD-H, AGP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2273 306 2259 MON-368 PO07-01 Monday 1928 2012


1924 ENDO12L_MON-369 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Does Insulin-Resistant Type 2 Diabetes Alter Vitamin D Status? Kamal AS Al-Shoumer, Vasanthy S Nair, Aida H Ali Faculty of Medicine, Kuwait University, Jabriyah, Kuwait; Mubarak Al-Kabeer Hospital, Jabriyah, Kuwait [bold]INTRODUCTION: [/bold][br]In the past several years, vitamin D has received an enormous renewal of attention, at various health levels. Recent literature suggests a link between vitamin D deficiency and several disorders including insulin resistant diabetes mellitus. Insulin resistance and impaired insulin action have also been found to be prevalent in vitamin D deficiency. Data on vitamin D status in Kuwaiti subjects with insulin resistant type 2 diabetes are not known.[br][bold]SUBJECTS [amp] METHODS:[/bold][br]We studied 69 Kuwaiti subjects with insulin resistant type 2 diabetes and 60 normal control matched for age and sex. Patients were included in the study if they are either treated with diet or oral antidiabetic agents, but not with insulin. After overnight fasting, blood was collected for the measurement of parameters of glycemia (glucose, insulin and HbA1c), parameters of calcium homeosatasis (corrected serum calcium, phosphate, alkaline phosphatase (ALP)), serum 25-hydroxyvitamin D (25OHD) and parathyroid hormone (PTH) level.[br][bold]RESULTS: [/bold][br]Patients had been diabetic for a median of 6 year (range, 0.5-20.5 years). They had significantly elevated fasting glucose (P=0.001), insulin (P=0.0003) and HbA1c (0.005) than controls. Patients and controls had similar levels of serum corrected calcium and ALP, whereas serum phosphate was significantly lower in the patients compared with controls (patients versus controls, mean [plusmn]SEM, 1.1[plusmn]0.02 vs. 1.23[plusmn]0.02 mmol/l, P= 0.0001). Patients and controls had similar 25OHD levels (25.4[plusmn]2.1 vs. 21.6[plusmn]2.0 nmol/l, P=0.09) and the levels in both were in the deficiency range ([lt]50nmol/l) compared with WHO reference values. PTH levels were similar in the patients and controls. 25OHD correlated negatively with PTH (r= - 0.4, P=0.02), but did not demonstrate any relation with fasting glucose, insulin, BMI, HbA1c, corrected ca, phosphate, ALP or duration of diabetes.[br][bold]CONCLUSION:[/bold][br]Kuwaiti subjects (diabetic and normal controls) had similar vitamin D concentrations and the levels were in the deficiency range. Insulin resistance type 2 diabetes does not seem to have an influence on vitamin D level in Kuwaiti subjects.[br][br]Nothing to Disclose: KASA-S, VSN, AHA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 219 306 2260 MON-369 PO07-01 Monday 1929 2012


1925 ENDO12L_MON-370 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) Declining UVB Irradiation and the Vitamin D Status of German Type 1 Diabetes Patients Johannes Langer, Marissa Penna-Martinez, Markus Wallasch, Hanns Ackermann, Klaus Badenhoop University Hospital Frankfurt am Main, Frankfurt am Main, Germany; German Federal Environmental Agency, Langen, Germany; University Hospital Frankfurt am Main, Frankfurt am Main, Germany [bold]Background:[/bold] Type 1 diabetes (T1D) is an autoimmune disease caused by the destruction of insulin-producing pancreatic beta cells. A possible risk factor for T1D are low cholecalciferol (25(OH)D[sub]3[/sub]) concentrations. The 25(OH)D[sub]3[/sub] concentration depends on alimentary intake but mostly on human skin production that is influenced by exposure to UVB radiation (290-315nm). Our aim was to investigate whether UVB radiation has changed in the past years and if this influences patients[apos] 25(OH)D[sub]3[/sub] concentrations. [bold]Methods:[/bold] We analysed the 25(OH)D[sub]3[/sub] concentration of 287 in- and outclinic patients (age 3917.83 years, male n = 137; female n = 150) with T1D from 2004 to 2007 at the University Hospital Frankfurt am Main. Additionally, daily UVB levels of this period were obtained by the German Federal Environmental Agency (Langen) for the Frankfurt region. Wilcoxon-Mann-Whitney test and Spearman[apos]s rho were used for statistical analyses. [bold]Results[/bold]: 25(OH)D[sub]3[/sub] levels of patients with T1D were in summer (Apr-Oct) significantly higher than in winter (Nov-Mar) season (17.0 ng/ml versus 13.0 ng/ml, p = 8 x 10[sup]-3[/sup]). Furthermore, the 25(OH)D[sub]3 [/sub]concentration of T1D patients obtained in summer 2004 to 2007, declined significantly (22.5 ng/ml versus 16.0 ng/ml; correlation coefficient = rho =-0.24, p = 2 x 10[sup]-3[/sup]) whereas it remained stable in winter (11.5 ng/ml versus 13.0 ng/ml; rho = -0.03, p = 0.73) during the same period. During this observation period, UVB radiation declined significantly in summer (rho = -0.21, p [lt] 10[sup]-6[/sup]) but no change was found in winter (rho = -0.07, p = 0.12). Finally, we detected a significant correlation between the 25(OH)D[sub]3[/sub] concentration and UVB radiation (rho = 0.56, p = 7 x 10[sup]-3[/sup]). [bold]Conclusion:[/bold] Our results suggest that decreasing UVB radiation in summer possibly due to changes in stratospheric ozone layers and caused by alteration of solar activity may lead to declining vitamin D levels in German patients with T1D.[br][br]Nothing to Disclose: JL, MP-M, MW, HA, KB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1759 306 2261 MON-370 PO07-01 Monday 1930 2012


1926 ENDO12L_MON-371 POSTER SESSION: Vitamin D Deficiency [amp] Metabolic Outcomes (1:30 PM-3:30 PM) 25-OH-Vitamin D and Procalcitonin Levels after Correction of Acute Hyperglycemia Nalan Metin Aksu, Duygu Yazgan Aksoy, Canan Akman, Hakki Yilmaz, Meltem Akkas, Bulent Okan Yildiz, Mehmet Mahir Ozmen, Aydan Usman Hacettepe University, Ankara, Turkey; Hacettepe University, Ankara, Turkey; Etlik Ihtisas Research and Training Hospital, Ankara, Turkey; Yildirim Beyazit Diskapi Research and Training Hospital, Ankara, Turkey [bold]BACKGROUND AND AIM: [/bold]Hyperglycemia is a frequently encountered complication of DM and in the absence of metabolic decompensation is a common finding in the Emergency Department (ED) We aimed to evaluate the 25 OH Vit D [25(OH)D] and procalcitonin (PCT) levels during hyperglycemia and after normalization of blood glucose.[br][bold]METHODS: [/bold]Eighty eight patients over the age of 18 years who presented with acute hyperglycemia to Hacettepe University Department of Emergency Medicine were included. Euglycemia was obtained within 6-12 hours and serum samples were taken from the patients on admission and 6 hours after normalization of blood glucose. Along with plasma glucose plasma 25(OH)D and PCT levels were measured using ELISA.[br][bold]RESULTS: [/bold]There were 88 (45 male) patients with a median age of 60.0[plusmn]13.9 years. Serum 25(OH)D levels increased in all patients after normalization of blood glucose and serum PCT levels decreased in whole group. This decrease was independent of type of diabetes or presence of infection.[br][bold]CONCLUSION:[/bold] We demonstrated an increase in 25(OH)D after normalization of blood glucose along with a decrease in PCT in patients with hyperglycemia. This effect was independent of the type of diabetes and presence of infection. Further studies are needed to evaluate the faster link between metabolic abnormalities, Vit D, PCT and inflammation.[br][br]Nothing to Disclose: NMA, DYA, CA, HY, MA, BOY, MMO, AU 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1132 306 2262 MON-371 PO07-01 Monday 1931 2012


1927 ENDO12L_MON-372 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) Analytical Performance Characteristics of Two New Automated Immunoassays for 25 Hydroxy Vitamin D Earle W Holmes, Jean Garbincius, Kathleen M McKenna Loyola University Stritch School of Medicine, Maywood, IL; Loyola University Medical Center, Maywood, IL A marked increase in the demand for 25 hydroxy vitamin D (25OHD) testing has led to the development of several new immunoassays for the measurement of 25OHD in un-extracted serum. We have compared 25OHD results measured by the Abbott Architect (ARC) and the Siemens Centaur2 (CT2) immunoassays with total 25OHD results determined by liquid chromatography/tandem mass spectrometry(LCMS) (Quest Diagnostics, Wood Dale, IL 60191). The study sample consisted of 163 randomly selected clinical specimens (123 f, median age: 54yr and 40 m, median age: 59 y) submitted for 25OHD testing between 3/2 and 3/7/11. 98 of the specimens contained only 25OHD3 and 65 contained both 25OHD3 and 25OHD2.[br]The results of linear regression analysis of the total 25 OHD results measured by each immunoassay (y) as compared to the total 25OHD (25OHD2+25OHD3) by LCMS (x) were as follows: ARC: r2=0.41; SE= 7.8ng/mL; slope=0.47+/-0.44; intercept= 11.4+/- 1.54 ng/mL.CT2: r2=0.76; SE= 8.7ng/mL; slope=1.12+/- 0.05; intercept= -8.3+/- 1.73 ng/mL. 33, 45, and 71 of the subjects in the study population had 25OHD results that indicated Vitamin D deficiency ([lt]20 ng/mL) when measured by LCMS, ARC, and CT2, respectively. The magnitudes of the absolute biases between the total 25 OHD results determined by LCMS and results determined by either of the immunoassays for the 25OHD2-containing specimens showed an under-recovery of 25OHD2 by the ARC and an over-recovery of 25OHD2 by the CT2. The differences between the total 25OHD results measured by LCMS and ARC or CT2 exceeded the maximum recommended total allowable error (+/- 25%) for 25OHD measurements in 40%, or 48%, respectively, of the clinical samples that were tested.[br]Our results showed that both of these new FDA-approved immunoassays were characterized by high degrees of random variability and significant constant biases relative to an established LCMS method, inaccurate measurement of 25OHD2, and a failure to meet even a minimum quality standard for analytical bias for many of the clinical specimens we tested. The inaccuracy of these immunoassays at 25OHD levels in the lower part of the analytical measuring range would have led to a marked overestimation of the prevalence of vitamin D deficiency in our study sample. The widespread use of new analytical methods with poor analytical quality may confound efforts to establish reference values for 25OHD in health and efforts to evaluate the role of vitamin D insufficiency as a risk factor in disease.[br][br]Nothing to Disclose: EWH, JG, KMM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1138 307 2263 MON-372 PO07-02 Monday 1932 2012


1928 ENDO12L_MON-373 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) Determinants of 25OHD[sub]3[/sub] Plasma Half-Life Kerry S Jones, Shima Assar, Ann Prentice, Inez Schoenmakers MRC International Nutrition Group, Fajara, Gambia; MRC Human Nutrition Research, Cambridge, UK Vitamin D status is usually assessed by measurement of the plasma concentration of 25-hydroxyvitamin D (25OHD) but this is a function of input from dietary intake and cutaneous synthesis and usage. We developed a method to measure the plasma half-life ([italic]t[sub]1/2[/sub][/italic]) of 25OHD[sub]3 [/sub](1) and investigated factors that may predict 25OHD[sub]3[/sub] [italic]t[sub]1/2[/sub][/italic] in 2 groups representing a broad range of known drivers of vitamin D metabolism.[br]Healthy adult males from Cambridge, UK and Keneba, The Gambia (each n=18) received an oral dose of [sup]2[/sup]H labelled 25OHD[sub]3[/sub] on day 1 (d1). Blood samples were collected before the dose on d1 and after on d6, 9, 21, 24, 27, 30 and 33. Tracer concentration was measured by UPLC-MS/MS. Plasma markers of vitamin D, calcium (Ca) and phosphate (P) metabolism and dietary Ca intake were measured. A series of univariant and multivariant models with stepwise backwards elimination of non-significant variables were used to examine the predictors of 25OHD[sub]3[/sub] [italic]t[sub]1/2[/sub][/italic]. A country*variable interaction term was used to allow for country differences. Predictors were selected on the basis of significance in univariant models and biological plausibility. Data are presented for countries combined.[br]Geometric mean (95% confidence interval) for biochemical parameters were: total 25OHD ([italic]t[/italic]25OHD) 42.5 (35.3-51.1) nmol/l; 1,25(OH)[sub]2[/sub]D 147.1 (134.3-161.0) pmol/l; parathyroid hormone (PTH) 40.6 (35.4-46.5) ng/l; plasma calcium corrected for albumin ([italic]p[/italic]Ca[sub]ALB[/sub]) 2.35 (2.33-2.37) mmol/l; plasma P ([italic]p[/italic]P) 1.04 (1.00-1.08) mmol/l. Means (SD) were: ionised Ca ([italic]i[/italic]Ca) 1.12 (0.03) mmol/L; Ca intake 722 (426) mg/d and 25OHD[sub]3[/sub] [italic]t[sub]1/2[/sub][/italic] 15.1 (3.1) d. Significant differences were found between countries in [italic]t[/italic]25OHD, 1,25(OH)[sub]2[/sub]D (both p[lt]0.0001), [italic]p[/italic]Ca (p 0.022), PTH (p 0.006) and Ca intake (p [lt]0.0001), but not [italic]p[/italic]P and 25OHD[sub]3[/sub] [italic]t[sub]1/2[/sub][/italic].[br]In univariant and multivariant analyses 25OHD[sub]3[/sub] [italic]t[sub]1/2[/sub][/italic] was consistently and significantly predicted by [italic]p[/italic]Ca[sub]ALB[/sub] (p 0.0272 and 0.0018), [italic]p[/italic]P (0.0364 and 0.0373) and PTH (0.0378 and 0.0276), after correction for country. [italic]t[sub]1/2[/sub][/italic] was not significantly predicted by [italic]t[/italic]25OHD, 1,25(OH)[sub]2[/sub]D, [italic]i[/italic]Ca or Ca intake. The association between [italic]t[sub]1/2[/sub][/italic] and both [italic]p[/italic]Ca[sub]ALB[/sub] and PTH was negative, and positive for [italic]p[/italic]P.[br]In conclusion, associations of 25OHD[sub]3[/sub] [italic]t[sub]1/2[/sub] [/italic]with PTH and [italic]p[/italic]P are consistent with current knowledge. The finding that [italic]p[/italic]Ca[sub]ALB[/sub] was negatively associated with 25OHD[sub]3[/sub] [italic]t[sub]1/[/sub][/italic][sub]2[/sub] and the absence of an association with [italic]i[/italic]Ca and Ca intake contrasts with current knowledge of determinants of vitamin D metabolism and requires further investigation.[br][br](1) Jones KS et al., Br J Nutr 2011 epub.[br][br]Sources of Research Support: UK Medical Research Council.[br][br]Nothing to Disclose: KSJ, SA, AP, IS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1432 307 2264 MON-373 PO07-02 Monday 1933 2012


1929 ENDO12L_MON-374 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) Method Agreement of New 25-OH Vitamin D Immunoassays vs LC-MS/MS Brian Keevil, Steven Lockhart, Laura Owen, Stefan Lorenz University Hospital of South Manchester, Manchester, UK; AbbottGmbH[amp]Co KG, Wiesbaden-Delkenheim, Germany [bold]Aim[/bold]. New automated immunoassays have been launched in 2011 to address the increasing demand of vitamin D (25(OH)D) testing. The new ARCHITECT 25-OH Vitamin D assay (Abbott) and the new Roche Vitamin D total assay were compared against LC-MS/MS.[br][bold]Methods[/bold]. Method agreement and correlation were evaluated on 300 specimens. The in-house LC-MS/MS was used as reference method. Recovery of 25(OH)D2 on the immunoassays was determined based on samples with high vitamin D2/D3 ratio ([gt]10) based on LC-MS/MS measurement.[br][bold]Results[/bold]. Passing-Bablok Correlation of Architect with LC-MS/MS was 0.90x + 1.16 (Pearson correlation = 0.89). Passing-Bablok Correlation of Roche with LC-MS/MS was 0.85x + 0.21 (Pearson correlation = 0.90). In the difference plot, results of the Roche assay dropped below the mean for values exceeding 60 ng/mL. This was not seen for the Architect assay. The mean recovery of 25(OH)D2 was 72.4% (95% CI: 58.6% to 86.2%) for Architect and 77.5% (95% CI: 63.1% to 91.8%) for Roche. The in-house LC-MS/MS was compared to other LC-MS/MS by assessment of DEQAS data and was found to read in average 10% higher than the LC-MS/MS mean of DEQAS participants (n = 15 DEQAS samples).[br][bold]Conclusions[/bold]. Both automated 25-OH Vitamin D immunoassays correlate well with LC-MS/MS and the vitamin D2 cross reactivities are acceptable and comparable.[br][br]Sources of Research Support: Study funded by AbbottGmbH[amp]Co.KG.[br][br]Disclosures: SL: Collaborator, Abbott Laboratories. Nothing to Disclose: BK, SL, LO 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 483 307 2265 MON-374 PO07-02 Monday 1934 2012


1930 ENDO12L_MON-375 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) A Novel Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS) Method for the Ultra-Sensitive Detection of 1,25-(OH)[sub]2[/sub] Vitamin D2 and D3 in Serum Samples Michael Jarvis, Adam Latawiec, Michal Weinstock AB SCIEX, Concord, Canada (For Research Use Only. Not for Use in Diagnostic Procedures.) Owing to the relatively poor ionization efficiency of 1,25-(OH)[sub]2[/sub]-Vitamin D3, and the very low concentration of this compound in plasma, the intrinsic sensitivity is quite low when this compound is measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The challenge is to improve the high sensitivity and selectivity of LC-MS/MS analysis, without resorting to chemical derivatization strategies, by using ultra-sensitive mass spectrometric detection.[br]In the work presented here, an ultra-sensitive LC-MS/MS method has been developed for the analysis of 1,25-(OH)[sub]2[/sub]-Vitamin D3 (DHVD) in serum. The method employs a 2D chromatographic separation followed by MRM detection of the DHVD species as their lithium adducts, which allows for low levels of detection without the use of derivatizing agents thereby greatly simplifying the analytical workflow. Online sample clean-up of the protein precipitated sample is accomplished using a POROS column, which minimizes the need for offline, manual sample preparation. Analytical chromatographic separation is accomplished using dual Phenomenex Onyx Monolithic C-18 columns to enhance the separation of complex biological matrices while providing extremely low back pressures. The flows in the preparative and analytical columns are regulated by the use of an auxilary Valco 10-port valve assembly.[br]Using a simple acetonitrile protein precipitation step followed by centrifugation, the method readily achieves a limit of detection of 10 pg/mL of 1,25-(OH)[sub]2[/sub]-Vitamin D3 in serum. To achieve this level of sensitivity, it was important to enhance the ionization efficiency of the target analyte by forming the lithium adduct, and to also use an ultra-sensitive tandem mass spectrometer. The linear dynamic range for the method ranged from 0 to 250 pg/mL in serum. The method has been demonstrated to be robust and reproducible, and the target analyte is effectively separated from other matrix componenents with minimal interferences in the ion source.[br][br]Disclosures: MJ: Employee, AB Sciex. AL: Employee, AB Sciex. MW: Employee, AB Sciex. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 969 307 2266 MON-375 PO07-02 Monday 1935 2012


1931 ENDO12L_MON-376 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) Integration of an Automated Sample Preparation Workstation for the Analysis of 25-OH Vitamin D by LC-MS/MS Michael Jarvis, Ruth Zhang, John Snider, Li Liu, Michal Weinstock AB SCIEX, Concord, Canada; Beckman Coulter, Indianapolis, IN (For Research Use Only. Not for Use in Diagnostic Procedures.) Liquid chromatography-tandem mass spectrometry technology provides laboratories with a powerful tool for robust, accurate, sensitive detection of a wide variety of analytes. However, in the absence of complete method automation, results are susceptible to human error at many different stages, including preparation of calibration standards, sample preparation, and data processing. Not only does method automation eliminate human error, thus increasing reproducibility, it also eliminates subjectivity during data processing, and furthermore has the potential to save a great deal of time.[br]In the work presented here, an LC-MS/MS method for the analysis of 25-OH Vitamin D2 and D3 has been developed, making use of commercially available plasma calibrators and controls. In addition to manual preparation, all steps of sample processing could be automated using a BioMek NX[sup]P[/sup] platform. The sample preparation consisted of a liquid-liquid extraction in hexane, followed by evaporation to dryness under N[sub]2[/sub] gas, and reconstitution in 50:50 v/v MeOH and H[sub]2[/sub]O.[br]Samples were loaded in test tube format and the final samples were prepared in a 96-well plate format, and then transferred to the LC-MS/MS system for analysis of 25-OH Vitamin D2 and D3. The LC-MS/MS data acquisition, processing, and reporting were performed using the Cliquid[sup][reg][/sup] software. Upon completion of data acquisition, the software automatically performed quantitation for the target analytes included in the method, and automatically generated and printed out reports. The reproducibility of the automated protocol versus manual protocol was assessed by preparing and analyzing replicates of each calibration standard. The measured CVs were at least equivalent between protocols over the entire concentration range covered by the assay. The method displayed good linearity for all analytes, with R[gt]0.999.[br][br]Disclosures: MJ: Employee, AB Sciex. RZ: Employee, Beckman Coulter. JS: Employee, Beckman Coulter. LL: Employee, Beckman Coulter. MW: Employee, AB Sciex. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2079 307 2267 MON-376 PO07-02 Monday 1936 2012


1932 ENDO12L_MON-377 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) 25 OH Vitamin D Testing by Three Methods: Do They Identify the Same Level of Deficiency or Sufficiency? Ana M Sequera, Gabriela Ruibal, Eleonora Nunez Chavarria, Hilda Farelo, Anabella Odriozola, Maria Fosatti, Delia Pelletier, Paula Esteban, Silvia Corti, Maria Aguet, Cecilia Aguirre, Fernanda Aseguinolaza, Gustavo Benitez, Sofia Coniglio, Erich Fradinger, Romina Fiorenzano, Ana Fontanazza, Maria Gonzalez, Nanci Gutierrez, Carolina Justo, Maria Lescurat, Marcela Miro, Silvana Montero, Hugo Molinas, Clelia Paredes, Adriana Pittau, Amelia Reverendo, Cintia Tarifa, Samuel Barcudi, Ana Canteli, Cristina Calderon, Gisella Di Prieto Reveand, Angel Galassi, Valeria Garcia, Julieta Gimenez, Laura Guerrero, Nadia Kogovsek, Gabriela Munoz, Marcela Niro, Gabriel Ramos, Mariana Rocca, Natalia Rossi, Cecilia Fenili, Mirta Gurkinkiel, Andrea Kozak Argentine Society for Endocrinology and Metabolism, Buenos Aires, Argentina Methods available for 25 OH Vitamin D (VIT D) can recognize both D2 and D3 (total VIT D) or only D3 and present different analytical characteristics. However, there[apos]s only one consensus value for VIT D sufficiency ([gt] 30 ng/ml) or deficiency ([lt] 20 ng/ml)[br]The aim of our work was to identify or evaluate degrees of VIT D deficiency during winter, comparing results tested with three available analytical methods.[br]Materials and Methods: 254 serum samples from healthy adults (18 - 45 years) without Vit D intake, were included. Subjects with severe organic insufficiencies, or those on medication for bone metabolism alterations or VIT D were excluded.[br]Samples were aliquoted and stored at -20[ordm]C during the study. VIT D was tested by three methods measuring total VIT D: radioimmunoassay (RIA,DIASORIN); chemiluminiscence (CLIA, ADVIA Centaur, Siemens Diagnostics), and electrochemiluminiscence (ECLIA, Cobas e411, Roche Diagnostics). Samples were divided into three geographical regions: North (NZ) (n:43) 26[deg]-27[deg] South latitude, mean temperature (T[deg]) 16[deg]C; Centre (CZ) (n:102), 31[deg]-36[deg] South latitude, mean T[deg] 9[deg]C; and South (SZ) (n:109) 38[deg]-51[deg] South latitude, mean T[deg] 3[deg]C. Results: % Samples with values of VIT D [le] 20 ng/ml (expressed median and range) were: in NZ, RIA 20.9 % samples, median 17 (10.0- 20.0) ng/ml; CLIA 45.5.% samples, median: 16.1 (13.0-19.2) ng/ml and ECLIA 16.7 % samples, median 15.9 (10.0-19.1) ng/ml. In CZ, RIA 35.2 %samples, median 15.5 (6.20- 20.0) ng/ml, CLIA 33.3.% samples, mean 18.1 (11.1-19.7) ng/ml and ECLIA 37.9 % samples, median 17.1 (8.4-19.8) ng/ml. In SZ, RIA 51.4 % samples, median 15.0 (7.00- 20.0) ng/ml, CLIA 67% samples, median 14.8 (8.4-20.0) ng/ml and ECLIA 42.2 % samples, mean 14.3 (3.00-19.6) ng/ml. Comparison between all samples with VIT D [le] 20 ng/ml by all systems showed significant differences between RIA and ECLIA and between CLIA and ECLIA (p[lt] 0.05, Friedman, Dunn test). Samples [ge] 30 ng/ml by RIA showed greater dispersion of values in other methods (p[lt] 0.05, RIA vs CLIA and RIA vs EQLIA Friedman, Dunn test). Bland-Altman analysis showed concordance between RIA and CLIA and RIA and ECLIA.[br]Conclusions: Levels of VIT D sufficiency or deficiency shouldn[apos]t be inferred from latitude or season, they must be measured. Analytical systems available do not characterize patients in the same way, so method-dependent cutoff levels should be determined or verified.[br][br]Nothing to Disclose: AMS, GR, ENC, HF, AO, MF, DP, PE, SC, MA, CA, FA, GB, SC, EF , RF, AF, MG, NG, CJ, ML, MM, SM, HM, CP, AP, AR, CT, SB, AC , CC, GDPR, AG, VG, JG, LG, NK, GM, MN, GR, MR, NR, CF, MG, AK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1109 307 2268 MON-377 PO07-02 Monday 1937 2012


1933 ENDO12L_MON-378 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) Clinical and Genetic Analysis of Patients with Vitamin D-Dependent Rickets Type 1: Evidence of a Non-[italic]CYP27B1[/italic] Encoded Vitamin D 1[alpha]-Hydroxylase? Yufei Shi, Minjing Zou, Roua A Al-Rijjal, Iffet Bircan, Sema Akcurin, Brian Meyer, Erdem Durmaz Akdeniz University School of Medicine, Antalya, Turkey; King Faisal Specialist Hospital [amp] Research Centre, Riyadh, Saudi Arabia Vitamin D-dependent rickets type 1 (VDDR-I), also known as pseudovitamin D deficiency rickets or vitamin D 1[alpha]-hydroxylase deficiency, is an autosomal recessive disorder characterized by early onset of rickets with hypocalcemia. Mutations in the [italic]CYP27B1 [/italic]gene, which encodes vitamin D 1[alpha]-hydroxylase, are the genetic basis of the disease. We investigated [italic]CYP27B1[/italic] mutation in 7 patients and their parents from 4 separate families by sequencing analysis of entire coding region of the [italic]CYP27B1[/italic]gene. All the patients had clinical and radiographic features of rickets with characteristic laboratory abnormalities of VDDR-I: hypocalcemia, hypophosphotemia, high serum alkaline phosphatase and parathyroid hormone (PTH), and low serum 1, 25-dihydroxyvitamin D[sub]3[/sub] [1, 25-(OH)[sub]2[/sub]D[sub]3[/sub]]. Sequencing analysis identified biallelic [italic]CYP27B1[/italic] mutations in all the patients and monoallelic mutations in their parents, indicating an autosomal recessive inheritance. One patient from the first family was compound heterozygous for c.1166G[gt]A (p.R389H in exon 7) and a novel nonsense mutation c.1079 C[gt]A, (p.S360X in exon 6). Two patients from the second family were homozygous for a novel splice donor site mutation in intron 1 (IVS1+2 T[gt]G) causing partial retention of the intron 1 and a shift in the reading frame. Both novel mutations lead to the complete loss of 1[alpha]-hydroxylase activity. Four patients from families 3 and 4 were homozygous for a previously reported insertion mutation (c.1325-1333 ins CCCACCC, P442fsX466 in exon 8), which is devoid of 1[alpha]-hydroxylase activity. Interestingly, one patient who was presented with severe hypocalcaemia and seizures at 4 months of age as a result of P442fsX466 mutation improved spontaneously since 11 years of age and does not need regular treatment. Her lab tests showed normal serum calcium and 1, 25(OH)[sub]2[/sub]D, elevated serum PTH and hypophosphotemia after refusing to take medication for 12 months. Her bone density scan (DEXA) was normal. However, her brother who is carrying the same mutation could not tolerate one week calcitriol withdrawal. These results indicate that although there is a good genotype-phenotype correlation in VDDR-I, some patients may recover at least partially from the loss of [italic]CYP27B1[/italic] function during puberty, probably due to 1[alpha]-hydroxylase activity exerted by another enzyme not encoded by [italic]CYP27B1[/italic] or compensation through epigenetic mechanisms.[br][br]Nothing to Disclose: YS, MZ, RAA-R, IB, SA, BM, ED 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 119 307 2269 MON-378 PO07-02 Monday 1938 2012


1934 ENDO12L_MON-379 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) Dose[ndash]Response Relationship between Vitamin D Supplementation and Calcium Absorption in Postmenopausal Women Vinod Yalamanchili, John Christopher Gallagher, Lynette M Smith Creighton University Medical Center, Omaha, NE; UNMC College of Public Health, Omaha, NE Background: 1,25 dihydroxyvitamin D (1,25 OH2D3) is the major hormone that controls intestinal calcium absorption. Serum 1,25 OH2D3 decreases when serum 25 hydroxyvitamin D (25OHD) falls below [lt]10ng/ml but it is not known at what serum 25OHD level malabsorption of calcium can occur. Based on cross sectional data, one study suggested that a threshold for normal calcium absorption occurred at a serum 25OHD level of 30 ng/ml but in another study it was 5-10ng/ml. We performed a longitudinal study of different doses of vitamin D on absorption to measure the effect on calcium absorption and look for a threshold effect.[br]Methods: 163 Caucasian women, ages 57-85 years, were randomized to different doses of vitamin D3 - 400, 800, 1600, 2400, 3200, 4000, 4800 IU/day or placebo. Calcium intake was increased to 1200-1400mg/day from an average of 691 mg/day. Main inclusion criteria was that subjects had serum 25OHD level [lt] 20ng/ml. Exclusion criteria were medical illness or medications known to affect calcium absorption. Calcium absorption was measured at baseline and after 12 months using a single isotope method with 100 mg calcium and 5 microcuries Ca45, serum Ca45 was corrected for weight or BMI. Serum 25OHD and 1,25 OH2D3 were measured by immunoassay (Diasorin). We used multivariate regression that included age, dietary calcium intake, weight, baseline and final calcium absorption and serum 25OHD.[br]Results: Mean baseline serum 25OHD was 15.6 ng/ml (39nmol/L) and increased as a quadratic function plateauing at 45ng/ml on the 4800IU dose. In the longitudinal study, calcium absorption increased on vitamin D but was more highly correlated with final serum 25OHD (p=0.009) than with vitamin D dose (p=0.033). The increase in calcium absorption was approximately 18 percent between serum 25 OHD levels of 15 to 60 ng/ml and there was no evidence of a plateau. In the baseline analysis serum 25OHD was divided into 5ng/ml groups between 3-20ng/ml and as serum 25OHD decreased below 5ng/ml there was a decrease in serum 1,25 OH2D3 but not in calcium absorption.[br]Conclusions: The results show that vitamin D increases calcium absorption. The association with vitamin D dose was inconsistent and absorption was more clearly related to the final serum 25OHD level. No threshold level of serum 25OHD for calcium absorption was found at baseline or longitudinally suggesting that the threshold for decreased calcium absorption occurs at a very low serum 25OHD [lt] 5ng/ml.[br][br]Sources of Research Support: National Institutes on Aging.[br][br]Nothing to Disclose: VY, JCG, LMS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1705 307 2270 MON-379 PO07-02 Monday 1939 2012


1935 ENDO12L_MON-380 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) The Effect of Vitamin D3 Supplementation on Glycemic Control in Type 2 Diabetic Patients Having Suboptimal Vitamin D Level, in an Open-Label, Randomized Controlled Trial Mansour Khater Alzahrani King Abdulaziz Medical City, Riyadh, Saudi Arabia Background: Recently, there is accumulating evidence to suggest that altered vitamin D and calcium homeostasis may play a role in the development of type 2 DM.[br]Objective: The aim of the current study was to evaluate and validate the effect of vitamin D supplementation on glycemic control in the form of Glycated hemoglobin and fasting blood sugar in type 2 DM.[br]Material and Methods: Randomized controlled clinical trial. The study trial passed through two phases. Phase one: screening of diabetic patients to estimate the prevalence of suboptimal vitamin D level. Phase two: random allocation of diabetic patients with suboptimal vitamin D level (detected in phase one) to either vitamin D3 supplement (intervention or treated group) or to no vitamin D3 supplement (control group).[br]Results: In the current study, 248 type 2 diabetic patients had been screened for vitamin D deficiency. The great majority of diabetic patients had suboptimal level of vitamin D (98.4%). Two hundred Diabetic patients out of 248 with suboptimal level of vitamin D were equally and randomly assigned to vitamin D supplementation group or control group. The duration of follow up was three months, from randomization till the end of the study. The comparison between different parameters before and after treatment with cholecalciferol in the treated group showed that vitamin D level had been increased significantly after treatment, from 24.96 nmol/L to 82.80 nmol/L), P[lt]0.001. Diastolic blood pressure has been decreased significantly after treatment, from 76.43 mm/hg to 73.25 mm/hg, p=0.021. There was no statistically significant difference in other compared parameters in the treated group, after three months of treatment with cholecalciferol.[br]Conclusions: The administration of vitamin D did not significantly improve type 2 DM indicators (glycosylated haemoglobin and fasting blood glucose), lipid profile of type 2 diabetic patients and their BMI in three months.[br][br]Sources of Research Support: King Abdullah International Medical Research Center; King Abdulaziz City for Science and Technology.[br][br]Nothing to Disclose: MKA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 181 307 2271 MON-380 PO07-02 Monday 1940 2012


1936 ENDO12L_MON-381 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) Vitamin D[sub]3[/sub][apos]s Effect on Calcium Absorption in Dialysis Patients Mohsen Zena, Laura Armas, Robert Heaney, Susan Dowell, Richard Lund, Lisa Moffatt, June Bierman, Karen Rafferty Creighton University Medical Center, Omaha, NE [bold]Background:[/bold] Vitamin D supplementation in the form of ergocalciferol or cholecalciferol has been a source of much interest in the general population as well as in patients with disease. The KDIGO guidelines make vitamin D supplementation recommendations for patients on dialysis, i.e. that they should be made vitamin D sufficient because of the need for 25(OH)D for use by other systems and cells in the body. While calcitriol or its analogues are widely used to replace systemic levels of 1,25(OH)[sub]2[/sub]D and native vitamin D has been shown to be effective in raising 25(OH)D levels in dialysis patients, little is known on the safety of vitamin D supplementation in dialysis patients or its effects on calcium absorption. We report here the effect of weekly oral 20,000 IU cholecalciferol for 12 weeks on calcium absorption.[br][bold]Study Design: [/bold]Prospective study with [ldquo]pre and post[rdquo] testing to evaluate native vitamin D supplementation on intestinal absorption of calcium in dialysis patients.[br][bold]Results: [/bold]Two calcium absorption tests were done using methods previously described by Heaney et al. One at baseline and the second after 12 weeks of 20,000 IU weekly of vitamin D[sub]3[/sub] supplementation. 25(OH)D, 1,25(OH)[sub]2[/sub]D, PTH, calcium, albumin, and phosphorus, were measured at baseline and [sim] biweekly throughout 12 weeks of Vitamin D[sub]3[/sub] treatment. 30 stage 5 CKD patients, requiring dialysis, completed the study [16 males, 14 females, mean age 54.5 yrs (SD 11.3)], mean height 169 cm (SD 9), mean weight 199 lbs (SD 56), and mean BMI 32 (SD 9). Race was self reported as 24 African American, 5 Caucasian, and 1 American Indian. Eight subjects had diabetes. Subjects had been on dialysis for an average of 5.4 yrs (SD 4.6). Reasons for dialysis were: 8 diabetes, 15 hypertension, and 7 other renal diseases. 23 subjects were on calcitriol analogues. Mean 5 hr absorption fraction was 13% (SD 8) at baseline and 12% (SD 7) after vitamin D supplementation. There was no change in calcium absorption between the 2 measurements. 25(OH)D levels were 16.8 ng/mL (SD 9.3) at baseline and 51.6 ng/mL (SD 15.8) after 12 weeks of treatment. This was significantly different (P [lt]0.001, Wilcoxon signed ranks test).[br][bold]Conclusion: [/bold]Supplementation with cholecalciferol at levels shown to increase calcium absorption in healthy adults has no effect on calcium absorption in patients on dialysis. The clinical implication is that risk of hypercalcemia is negligible in dialysis subjects supplemented with cholecalciferol.[br][br]Nothing to Disclose: MZ, LA, RH, SD, RL, LM, JB, KR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 892 307 2272 MON-381 PO07-02 Monday 1941 2012


1937 ENDO12L_MON-382 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) Vitamin D Binding Protein Levels in Female Patients with Primary Hyperparathyroidism X Wang, SA Shapse, S Wei, D Sukumar, J Ghosh RWJMS-UMDNJ, New Brunswick, NJ; Rutgers University, New Brunswick, NJ [bold]Introduction:[/bold] Vitamin D deficiency is common in patients with primary hyperparathyroidism (PHPT) but the mechanisms of vitamin D deficiency remains unclear. Vitamin D binding protein (VDBP) is the major transport protein for vitamin D metabolites in plasma, and approximately 88% of 25-hydroxyvitamin D (25OHD) in the circulation is bound to VDBP. It has recently been suggested that urinary loss of VDBP in patients with type 1 DM may contribute to their low levels of serum 25OHD. We therefore hypothesized that low levels of VDPB might also contribute to the apparent 25 OHD deficiency in PHPT patients.[br][bold]Subjects and Methods[/bold]: Twenty-five female patients with PHPT and twenty five age and BMI-matched healthy female subjects were enrolled in the study. PHPT was diagnosed in patients with both elevated serum calcium ([gt]10.2 mg/dl) and iPTH ([gt]66 pg/ml) levels. Both controls and PHPT patients were free of liver and renal diseases. VDBP was assayed in serum sample by ELISA. Data is expressed as mean [plusmn] SD.[br][bold]Results[/bold]: There is no significant difference in age (55.5[plusmn]8.8 [italic]vs[/italic] 56.4[plusmn]9.0) and BMI (31.0[plusmn]6.0 vs 32.3[plusmn]8.8 kg/m[sup]2[/sup]) between the two groups (P[gt]0.05). There is significant difference in calcium (10.8[plusmn]0.5 [italic]vs[/italic] 9.4[plusmn]0.7mg/dl) and iPTH (133[plusmn]82 [italic]vs[/italic] 36[plusmn]4.6 pg/ml) between the two groups (P[lt]0.001). 25OHD is lower in female PHPT patients (19.5[plusmn]7.0 ng/ml) than normal controls (25.2[plusmn]7.8 ng/ml, P =0.0109). VDBP in female PHPT patients is significantly lower compared to control subjects (36.1[plusmn]4.6 mg/dl [italic]vs [/italic]41.0[plusmn]3.2mg/dl P[lt]0.001). VDBP inversely correlates with PTH (r=-0.29), Ca (r = -0.43) (p [lt] 0.05) and shows a positive trend with 25OHD (p= 0.055).[br][bold]Conclusion and discussion[/bold]: Both serum 25OH D and VDBP levels were lower in female patients with PHPT compared to controls. We suggest that the low VDBP levels might be one of the mechanisms contributing to low vitamin D levels in female PHPT patients. The etiology of the decreased VDBP and its relationship to PTH and 25OHD levels require further investigation.[br][br]Nothing to Disclose: XW, SAS, SW, DS, JG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1961 307 2273 MON-382 PO07-02 Monday 1942 2012


1938 ENDO12L_MON-383 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) Variations in Parathyroid Hormone Concentrations in Patients with Low Vitamin D Bhanu Teja Paturi, Amal Shibli Rahhal University of Iowa Hospitals and Clinics, Iowa City, IA In Vitamin D deficiency parathyroid hormone starts increasing when the 25 hydroxyvitamin D (25OHD) drops below 30 ng/dl and PTH is often used to define vitamin D sufficiency. However in clinical practice, we have observed that the PTH is not always elevated in patients with low vitamin D levels and that the time to PTH normalization after correction of vitamin D deficiency varies widely between patients. The objective of this study was to examine variations in PTH levels in patients with low vitamin D levels and to examine how PTH levels changed following correction of the vitamin D deficiency.[br]We conducted a retrospective cohort study of all adults (age [ge] 18 years) with a 25OHD level [lt] 30 ng/ml seen in our clinic at the University of Iowa over a four year period. We excluded patients with no available baseline PTH values, as well as patients with concomitant conditions that may be associated with abnormal PTH levels, including abnormal kidney function (GFR[lt]60), hypercalcemia, hyperparathyroidism, hypoparathyroidism and rickets. We then examined variability in baseline PTH levels and the changes in PTH after correction of the vitamin D deficiency/insufficiency. The mean age of our cohort was 55.5 years, mean daily calcium intake was 1002.8 mg, 25OHD- 20.5 ng/ml, PTH-59.8 pg/ml. After treatment with vitamin D (average duration 10 months), the mean 25OHD increased from 20.5 to 40.7 ng/ml (p [lt] 0.01) and the mean PTH decreased from 59.8 to 56.2 pg/ml (p 0.04). 73 patients (70% of the cohort) had a normal PTH concentration at baseline. Patients with a normal baseline PTH did not experience a significant change in their PTH after normalization of the 25OHD. However, in patients with an elevated baseline PTH, the PTH level decreased from 103.3 to 72.3 pg/ml after normalization of the 25 OHD (p 0.03). Of the 31 patients who had an elevated PTH at baseline, follow up data was available on 22. Only10 patients (45.5%) experienced immediate normalization of the PTH as their 25OHD increased above 30 ng/dl, while the PTH remained elevated in the rest for up to 2 years.In this group of patients with a 25OHD concentration [lt] 30 ng/dl, up to 70% had a normal baseline PTH level. In patients with an elevated baseline PTH, the hyperparathyroidism seems to [ldquo]lag behind[rdquo] correction of the vitamin D status in more than 50 % of the patients. These findings suggest that PTH may not be helpful in defining vitamin D [ldquo]sufficiency[rdquo] and guiding vitamin D therapy in clinical practice.[br][br]Nothing to Disclose: BTP, ASR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 265 307 2274 MON-383 PO07-02 Monday 1943 2012


1939 ENDO12L_MON-384 POSTER SESSION: Vitamin D: Assays, Clinical Trials [amp] Physiologic Functions (1:30 PM-3:30 PM) Association between Vitamin D Receptor Polymorphisms and Haplotypes with 25-Hydroxyvitamin D Levels in Girls from South Brazil Betania Rodrigues Santos, Luis Paulo Gomes Mascarenhas, Fabiola Satler, Margaret Cristina da Silva Boguszewski, Poli Mara Spritzer Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil; Universidade Federal do Paran[aacute], Curitiba, Brazil; Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil Vitamin D deficiency is highly prevalent in some pediatric and adolescent populations. Vitamin D receptor (VDR) gene polymorphisms have been associated with vitamin D levels. We determined the prevalence of vitamin D deficiency in girls from southern Brazil and investigated whether the genotypic distribution of the BsmI, ApaI and TaqI polymorphisms of the VDR gene and their haplotypes were associated with vitamin D levels. Cross-sectional study including 234 apparently healthy girls aged 7 to 18 years. Height and weight were measured for calculation of body mass index (BMI) percentiles for age. The subjects were classified as normal weight (BMI [lt] 85 percentile), overweight (85 percentile [le] BMI [le] percentile 95) or obese (BMI [gt] 95 percentile). Plasma levels of 25-hydroxyvitamin D [25(OH)D] were assessed and 25(OH)D status was classified as sufficient ([ge] 30 ng/mL), insufficient (20-29 ng/mL) or deficient ([lt] 20 ng/mL). Participants were genotyped for ApaI (rs7975232), TaqI (rs731236) SNPs, both by polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) analysis, and BsmI (rs1544410) SNP, by real-time PCR with allelic discrimination assays. The median BMI percentile was 61.96 (33.29-84.99). The frequency of overweight/obesity was 24.9%. Circulating levels of 25(OH)D ([ge] 30 ng/mL) were adequate in 9.4%; insufficient in 54.3% (20-29 ng/mL); and deficient in 36.3% ([lt] 20 ng/mL). Genotype frequencies were GG = 47.0%, GA = 41.5%, and AA = 11.5% for BsmI; GG = 16.7%, GT = 52.6%, and TT = 30.8% for ApaI; TT = 46.2%, TC = 44.9% and CC = 9.0% for TaqI. The BsmI (G[rarr]A) polymorphism was in partial linkage disequilibrium with the ApaI (G[rarr]T) polymorphism (|D[apos]| = 0.964; r[sup]2[/sup]=0.330), and in almost complete linkage disequilibrium with the TaqI (T[rarr]C) polymorphism (|D[apos]| = 0.919; r[sup]2[/sup] =0.807). The ApaI (G[rarr]T) polymorphism was also in partial linkage disequilibrium with the TaqI (T[rarr]C) polymorphism (|D[apos]| = 0.970; r[sup]2[/sup]=0.319). The presence of the wild genotype of BsmI, ApaI and TaqI SNPs and the GGT haplotype were significantly associated with lower 25(OH)D levels, even after adjustment for season at the time of the blood collection, BMI percentiles and age. 25-hydroxyvitamin D deficiency and insufficiency were highly prevalent in this sample. The BsmI, ApaI and TaqI wild variants of the VDR gene, as well as the GGT haplotype, were associated with lower vitamin D levels.[br][br]Sources of Research Support: Brazilian National Institute of Hormones and Women[apos]s Health.[br][br]Nothing to Disclose: BRS, LPGM, FS, MCdSB, PMS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 216 307 2275 MON-384 PO07-02 Monday 1944 2012


1940 ENDO12L_MON-386 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Atypical Cause of Hoarseness in a Patient with Thyroid Nodules Manchin Chang, Hosai N Hesham, Edina Paal, Shikha Khosla, Eric Nylen Veterans Affairs Medical Center, Washington, DC; George Washington University, Washington, DC; Veterans Affairs Medical Center, Washington, DC; Veterans Affairs Medical Center, Washington, DC [bold]Background[/bold]: Hoarseness is an ominous symptom in patients with suspicion of thyroid malignancy(1-2). We present a case of a female with benign bilateral thyroid nodules presenting with persistent hoarseness.[br][bold]Case[/bold]: A female patient presented with hoarseness of several months duration and thyroid nodules found on CT scan, later confirmed by ultrasound. A fine needle aspiration revealed adenomatoid nodules. Persistent hoarseness prompted an ENT evaluation, revealing right vocal cord paralysis (VCP). MRI and a CT were consistent with right VCP. Surgical biopsy of the vocal cords showed stromal edema with subtle evidence of increased hyaluronic acid in the lamina propria and chronic inflammation that was predominantly perivascular and involving minor salivary glands. Focal venulitis was also noted. The epithelium showed no abnormality. In view of the histology and family history of systemic lupus erythematous (SLE), targeted laboratory evaluation was done and showed strong positivity for ANA, ds DNA, SM, and RNP (negative C3, SSA, SSb and anticardiolipin antibodies). The diagnosis of SLE was suggested. The patient was started on corticosteroids and plaquenil with improvement of symptoms. A laryngeal stroboscopic study at two months post-treatment showed persistent paralysis of the right vocal cord.[br][bold]Discussion[/bold]: Isolated laryngeal involvement of SLE is a rare finding. VCP also has been described in cases of drug induced lupus(3). Cases of bilateral VCP, can present with severe airway obstruction(4). Though histologic evaluation may be performed, in suspected SLE, specific histopathologic changes have not been reported. However, vasculitis, when present, may suggest the diagnosis of SLE. Laryngeal electromyography evaluation has shown denervation and reinnervation(5). There is little guidance in the literature directing the treatment of VCP(6), although it has been reported that improvement can be seen with systemic corticosteroids(7). This case highlights the importance of thorough evaluation of VCP in the setting of thyroid nodule(s). Although a thyroid malignancy is often suspected, the presence of a benign thyroid biopsy does not preclude the need for further evaluation, especially if there are still concerning symptoms.[br][br](1)Sugitani I et al., Endocrine Journal 1999; 46(1), 209-216. (2)Randolph GW et al., Surgery 2006; 139(3):357-62. (3)Hari CK et al., J Laryngol Otol 1998;112(9):875-7. (4)Karim A et al., Chest 2002; 121(3):990-3. (5)Lee JH et al., Am J Phys Med Rehabil 2008; 87(1):68-70. (6)Hughes M et al., Mod Rheumatol 2009; 19:441-442. (7)Ozcan KM et al., J Clin Rheumatol 2007; 13:278-279.[br][br]Nothing to Disclose: MC, HNH, EP, SK, EN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 650 308 2276 MON-386 PO56-03 Monday 1945 2012


1941 ENDO12L_MON-387 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Recurrent Goiter as a First Manifestation of Malignant Melanoma Marek Ruchala, Ewelina Szczepanek-Parulska, Katarzyna Piatek, Przemyslaw Majewski, Rafal Czepczynski, Jerzy Sowinski Poznan University of Medical Sciences, Poznan, Poland; Poznan University of Medical Sciences, Poznan, Poland Metastases to the thyroid are rare findings and gain little attention in the diagnostics of thyroid nodules. The most frequently reported primary sites are clear cell renal, bronchial, and breast carcinoma. Herewith we report a case of a patient with thyroid metastasis from malignant melanoma (MM), which was the first manifestation of neoplastic disease.[br]54-year-old woman was admitted to the endocrine outpatient clinic due to recurrent goiter and suspicion of anaplastic cancer on the basis of recent fine-needle aspiration biopsy (FNAB) results. She noticed a growing lump on the neck 3 months before. The lesion in the right thyroid bed was 1.5 cm in size, firm and highlighted the dermal layer. Patient[apos]s history revealed subtotal thyroidectomy due to nodular goiter 14 months earlier. The histopathological diagnosis was follicular adenoma. Ultrasound examination revealed hypoechogenic lesion of irregular borders in the right thyroid bed as well as enlarged nuchal lymph nodes. Sonoelastographic examination demonstrated decreased elasticity of the lesion and lymph nodes. Thyroid scintiscan revealed no Tc-99m uptake in the right thyroid lobe. During sonographic examination, a brownish skin lesion was noticed in the hairy skin of the head in the left temporal area. The dermatologists suggested the possibility of MM. FNAB was repeated. The results confirmed the diagnosis of MM metastases to the thyroid and lymph nodes. The biopsy of the cutaneous lesion was nondiagnostic. The PET scan indicated multiple foci of increased uptake in the whole body, suggesting advanced metastatic disease. The patient died within 2 months due to massive brain metastases.[br]Our report and review of the literature suggests that in any patient with a history or suspicion of non-thyroidal malignancy, a new thyroid mass might indicate metastasis. FNAB may help to discriminate between thyroid primary and metastatic neoplasms. Preoperative diagnosis of the metastasis and staging of the neoplastic disease helps to avoid unnecessary thyroidectomy. The MM metastases to the thyroid:[br]1. are rarely isolated and often constitute an evidence of disseminated disease[br]2. might be the first symptom and lead to diagnosis of MM[br]3. are indicative of a poor long-term survival[br]4. on cytology may mimic primary thyroid anaplastic cancer, if no suggestion about the MM suspicion is given to the pathologist[br]5. were rarely reported in the literature and our patient constitutes the 18th described case.[br][br]Nothing to Disclose: MR, ES-P, KP, PM, RC, JS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1511 308 2277 MON-387 PO56-03 Monday 1946 2012


1942 ENDO12L_MON-388 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) A Patient with Pendred Syndrome, High Thyroglobulin, and High I-131 Uptake after Total Thyroidectomy Elizabeth A Sedlis Singer, Nidhi R Narayan, Kent P Friedman, Susan B Zweig New York University Langone Medical Center, New York, NY; New York University Langone Medical Center, New York, NY [underline]Background[/underline]: Pendred syndrome (PS) is an autosomal recessive disorder characterized by hypothyroidism, goiter, partial defect in organification of iodide, and sensorineural deafness (1). A few studies have shown high thyrocyte iodide uptake in PS patients (1,2). We present a case of a PS patient with a very elevated thyroglobulin (Tg) and I-131 uptake after total thyroidectomy. [underline]Case[/underline]: A 26 year old woman with PS, diagnosed with hypothyroidism at age 6, was treated with levothyroxine until age 14. At age 25, an enlarging goiter caused dysphagia, and a total thyroidectomy was performed. Pre-op labs showed a TSH 1.6 mIU/L (0.4-4.5 mIU/L), free thyroxine (FT4) 0.6ng/dl (0.8-1.8ng/dl), FT4 by dialysis 0.8ng/dl (1.1-2.4ng/dl), total T4 4.3mcg/dl (4.8-11mcg/dl), thyroperoxidase (TPO) and Tg antibody (Ab) negative. Neck ultrasound (US) revealed an enlarged, hyperemic, and multinodular thyroid without discrete nodules. Pathology showed 3 foci of micro-papillary thyroid carcinoma. One month post-op, TSH was elevated (86 mIU/L) and Tg was very elevated (204 ng/ml) with negative Tg antibody. Neck US showed no remnant tissue; however withdrawal whole body scan (WBS) showed multifocal thyroid bed activity with 12% uptake. To aid in future monitoring, the patient received 75mCi of I-131. Post-therapy WBS revealed no change from prior and levothyroxine was started. [underline]Discussion[/underline]: PS is caused by mutations in the gene SLC26A4 which codes for pendrin, a glycoprotein involved in ion transport (3). In the thyroid, pendrin is located on the apical membrane of thyrocytes, and is thought to regulate the efflux of iodide into the follicular lumen (3). Mutations in the gene lead to a partial defect in iodide organification. Detection of the organification defect is done by performing a perchlorate test (1). Extent of goiter and hypothyroidism vary and tend to occur if nutritional iodide intake is low (3). Treatment of PS may include thyroid hormone replacement or surgery for symptomatic goiter. [underline]Conclusion:[/underline] We describe a PS patient with an incidentally found papillary thyroid cancer. Although no remnant thyroid tissue was seen on post-op US, the patient had elevated Tg levels and high I-131uptake in the neck on post-therapy WBS. We postulate this occurred because a small amount of residual thyroid tissue containing an organification defect, due to a mutation in pendrin, was stimulated by high TSH, leading to elevated Tg and an increased amount of iodide within the remaining thyrocytes.[br][br](1)Asakura Y et al., Am J Med Genet Part A. 2010; 152A:1793-7. (2)Palos F et al., J Clin Endocrinol Metab. 2008; 93:267-77. (3)Bizhanova A et al., Mol Cell Endocrinol. 2010; 322: 83-90.[br][br]Nothing to Disclose: EASS, NRN, KPF, SBZ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 583 308 2278 MON-388 PO56-03 Monday 1947 2012


1943 ENDO12L_MON-389 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) EEG Changes Due to Steroid Treatment of Hashimoto Encephalitis Omar M Salim, Khalid M Salim, Ralph DeSilva, Himat Pandy, Mohamad M Horani Mercy Gilbert Medical Center, Gilbert, AZ; Alsham Endocrinology, Chandler, AZ A 67 year old female was hospitalized for worsening confusion and memory loss over the past 2-3 weeks. She experienced episodes of poor comprehension, slow reaction time, sequences of unawareness and doing nonsensical activitiies.[br]The patient[apos]s past medical history includes hypertension, diabetes, and hypothyroidism. TSH levels have been elevated in the past.[br]MRI revealed two areas of periventricular white matter disease in subcortical distribution, though these areas appear old.[br]A preliminary EEG revealed that background activity was disrupted. Triphasic waves were seen, suggestive of metabolic encephalopathy. Partieto-occipital sharps were seen infrequently.[br]She was discharged after a spinal tap which showed normal WBC levels and elevated CSF protein 102 l (15 - 60 mg/100 mL). Her TSH levels were high at 38.6 (0.4 - 4 [mu]IU/mL). Her anti-thyroglobulin antibody titers were very high, 1032 ([lt] 20 IU/mL). Considering the acute confusion, abnormal EEG, elevated CSF protein, and abnormal anti-thyroglobulin antibody, the patient was treated with high dose oral steroids (prednisone, 60 mg/day).[br]Almost a month later, a second EEG was taken on suspicion of Hashimoto[apos]s encephalitis. Background activity was well-formed. Prior sharps were absent. No persistent asymmetry was seen. Overall, background activity significantly improved. Encephalopathy was completely resolved and background activity was within normal range. There was a remarkable improvement in cognition and function.[br]Hashimoto[apos]s encephalitis is an idiopathic neurological disorder associated with thyroid autoimmunity. It usually occurs in the 5th decade of life and in a subacute type possibly associated with cognitive dysfunction, inappropriate behavior, or memory loss. Clues to this diagnosis include elevated antithyroid antibodies (independent of thyroid status), abnormal TSH values, and exclusion of other causes of acute mental status changes. MRIs are generally normal. Nonspecific EEG abnormalities are seen in 90-98% of patients. After glucocorticoid treatment, a rapid reversal in patient[apos]s confusion and neurological debility can be affected.[br]Hashimoto[apos]s encephalitis should be considered in a patient with autoimmune thyroid disease accompanied with neuropsychiatric symptoms. There are generally normal MRIs. EEG characteristics usually have nonspecific abnormalities. Including it as a differential diagnosis is vital as glucocorticoid treatment could easily alleviate the mental status afflictions.[br][br]Nothing to Disclose: OMS, KMS, RDS, HP, MMH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 31 308 2279 MON-389 PO56-03 Monday 1948 2012


1944 ENDO12L_MON-390 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Is There a Role for Glucocorticoid Therapy in the Treatment of Acute Suppurative Thyroiditis (AST)? Deepa Mittal, Mrinalini Kulkarni-Date, Aneesh George Seton Family of Hospitals, Austin, TX Background: The thyroid gland is relatively resistant to infection, thus acute suppurative thyroiditis is rare. Traditional treatment has included antibiotics, surgical drainage, or thyroidectomy. The role of glucocorticoid therapy has not been clearly described.[br]Clinical Case: A 52 year -old man with history of non small cell lung carcinoma on palliative chemotherapy presented with rapid onset of neck swelling and pain. He was afebrile with a diffusely enlarged and tender thyroid gland. Laboratory data showed white cell count of 35K (4.5-11 K), ESR 112 mm/hr (0-20mm/hr), normal TSH, free T4, and subsequent blood cultures were negative. CT scan of the neck revealed diffuse heterogenous thyroid enlargement, greater in right lobe than left, and edema of anterior neck tissue, retropharyngeal space and carotid sheath. No focal abscess was identified on CT or ultrasound. ENT consultation was obtained, and a flexible laryngoscopy revealed inflammation of the retropharyngeal soft tissue, epiglottis, false cord and arytenoids. The patient was treated with IV glucocorticoids and IV broad-spectrum antibiotics. Glucocorticoids were discontinued on hospital day five and he was discharged on oral antibiotics on day six. The patient, however, returned the next day for increasing neck pain and swelling. He underwent ultrasound guided thyroid FNA and was again started on IV antibiotics and IV steroids. FNA culture grew methicillin-sensitive staph aureus, and pathology showed innumerable neutrophills. Based on the acuity of presentation, culture data, and pathology report, a diagnosis of AST was made.[br]A barium swallow study done to evaluate for occult piriform sinus tract as a cause of infective thyroiditis was negative. The patient was treated for a total of 21 days of antibiotics which included 14 days of IV antibiotics, and glucocorticoids were tapered over 21days. No surgical drainage was performed as there was no evidence of abscess. The patient had complete resolution of his thyroiditis.[br]Discussion: Though rare, a diagnosis of AST must be considered in presentation of anterior neck pain and swelling in the setting of normal thyroid function. The treatment approach of AST has been based on small case series and expert opinions, with few reports or data on the use of glucocorticoids. Our case will add to the scant literature on the possible role of glucocorticoid therapy in treatment of AST.[br][br]Paes et al. Acute Bacterial Suppurative Thryoiditis: A Clinical Review and Expert Opinion. Thyroid. 2010 Mar;20(3):247-55.[br][br]Nothing to Disclose: DM, MK-D, AG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2225 308 2280 MON-390 PO56-03 Monday 1949 2012


1945 ENDO12L_MON-391 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Dramatic Elevation of Serum Calcitonin Levels with Calcium Loading Test in C-Cell Hyperplasia Sujani Poonuru, Brian G Rose Medical College of Wisconsin, Milwaukee, WI [bold]Background:[/bold] Serum calcitonin (CT) is a very sensitive marker of medullary thyroid carcinoma (MTC). There are no well established serum CT cutoffs for distinguishing between MTC and C-cell hyperplasia (CCH), which is considered a preneoplastic lesion in familial forms of MTC.[br][bold]Clinical Case:[/bold] A 75-year old Caucasian male was referred to endocrinology clinic for an elevated serum CT level of 55.1 pg/ml (0.0-8.4 pg/ml). CT was measured by his dermatologist when he presented with intrascapular pruritis secondary to concerns of possible nostalgia paresthetica. He was otherwise asymptomatic. His past medical history was significant for hypertension and obstructive sleep apnea. He denied being on a proton pump inhibitor. He was a nonsmoker. Family history was negative for multiple endocrine neoplasia-2A and nostalgia paresthetica. Physical examination was unremarkable. Ultrasonogram of the thyroid showed a 5mm ovoid, solid, hypoechoic nodule without calcification in the superior left lobe. The lobes were otherwise homogenous. Fine needle aspiration of the nodule was unsatisfactory for interpretation due to acellularity. A needle rinse of the biopsy for CT was not properly processed. His RET proto-oncogene was negative. Calcium stimulated CT test was performed by infusion of calcium gluconate (2.5 mg/kg) over 3 minutes. During the infusion, he experienced transient nausea and hypotension. An EKG showed new onset atrial fibrillation. The baseline CT was 42.6 pg/ml and increased to 1111 pg/ml, 1240 pg/ml, 1070 pg/ml and 971pg/ml at 1, 3, 5 and 10 minutes respectively. Subsequently a total thyroidectomy and central neck dissection was performed and surgical pathology was reported as nodular CCH of the thyroid gland, and 5 of 5 lymph nodes were negative for malignancy. He remains in atrial fibrillation 7 months post calcium infusion.[br][bold]Conclusion: [/bold]The patient had a dramatic 29 fold increase from baseline to peak calcitonin via standard intravenous calcium infusion. Previous studies support a significant risk of MTC with elevated basal CT levels. Pentagastrin stimulated CT of greater than 100 pg/ml has been recommended as a cut-off to perform thyroid surgery. Levels of greater than 1000 pg/ml in the Constante et al series had a positive predictive value for diagnosis of MTC of 100% (1). Calcium stimulation may lead to greater CT stimulation than pentagastrin. There are no clear recommendations with respect to cut-offs for calcium stimulated CT.[br][br]1.Giuseppe Costante et al., J Clin Endocrinol.2007; 92(2):450[ndash]455.[br][br]Nothing to Disclose: SP, BGR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 122 308 2281 MON-391 PO56-03 Monday 1950 2012


1946 ENDO12L_MON-392 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Paraneoplastic Hyperthyroidism in a 34-Year-Old Man with Metastatic Testicular Choriocarcinoma Danielle H Lewis, Bobby Huda, Paul Carroll Guy[apos]s and St Thomas[apos] Hospitals NHS Foundation Trust, London, UK Elevated [beta]HCG can cause hyperthyroidism due to the ability of [beta]HCG to stimulate TSH receptors. In pregnancy, this phenomenon can result in hyperemesis gravidarum and thyrotoxicosis. Rarely some malignancies can produce excessive [beta]HCG.[br]A 34 year old man presented with a month history of dyspnoea and haemoptysis. There was an episode of pleurisy 2 months previously while abroad. One month previously there was some haemoptysis while working at high altitude. He also had minor lumbar back pain, constipation and minimal weight and appetite loss. There was no past medical or drug history. Examination was unremarkable except for bilateral gynaecomastia and lower back tenderness. He was clinically euthryoid.[br]Chest xray revealed multiple pulmonary metastases and testicular ultrasound showed features suggestive of seminomas. Diagnosis of probable testicular choriocarcinoma was made and his care transferred to our hospital.[br]Blood tests showed TSH[lt]0.01mlU/L, FT4 31.6pmol/L, FT3 9.2pmol/L. [beta]HCG was 777,289IU/L (alpha subunit 59.40IU/L), LDH 1820IU/L and Alpha foetoprotein 214.2klU/L. These results continued to climb before the commencement of chemotherapy to TSH[lt]0.01mlU/L, FT4 72.3pmol/L, FT3 15.7pmol/L. [beta]HCG peaked at 1,547,070IU/L and LDH 1909IU/L. Abdominal CT scan showed liver and lung metastasis and left para-aortic soft tissue mass invading retroperitoneal vascular structures. Thyroid antibodies were negative.[br]Chemotherapy (but no anti-thyroid medication) was commenced. He remained clinically euthryoid throughout treatment. There was a dramatic reduction in [beta]HCG with a corresponding reduction in free thyroid hormones. He received 3 courses of chemotherapy which initially showed good [beta]HCG and thyroid hormone response. However,[beta]HCG levels began to rise and he developed brain metastases. His care was transferred to another hospital where he is currently undergoing high dose chemotherapy and stem cell transplant.[br]Paraneoplastic hyperthyroidism is a rare but recognised complication of germ cell tumours. High levels of [beta]HCG act on TSH receptors stimulating excess thyroid hormone production. It is important to recognise that anti-thyroid medication is seldom required. Tumour specific chemotherapy reduces [beta]HCG production and, consequently there is less stimulation of thyroid hormone receptors and reduction in thyroid hormone synthesis. Interestingly, free thyroid hormones (as well as [beta]HCG) can act as tumour markers for progression of the malignancy.[br][br]Nothing to Disclose: DHL, BH, PC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1297 308 2282 MON-392 PO56-03 Monday 1951 2012


1947 ENDO12L_MON-393 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Nonautoimmune Hyperthyroidism in a Family with Gain-of-Function [italic] TSHR [/italic] Gene Mutation Caecilie Larsen, Lefkothea P Karaviti, Roy E Weiss, Alexandra M Dumitrescu Univ of Chicago, Chicago, IL; Univ of Chicago, Chicago, IL; Texas Children[apos]s Hospital, Houston, TX Nonautoimmune hyperthyroidism due to germline gain of function mutations in the [italic] thyrotropin receptor (TSHR) [/italic] gene is a rare condition. To date, only 32 cases have been reported. Among them, 20 had familial nonautoimmune hyperthyroidism.[br]The propositus, a 4.5 year old boy, presented with low body weight, difficulty gaining weight and hyperdynamic precordium, without obvious goiter. His mother underwent radioiodide treatment at age 18 years, followed 6 months later by thyroidectomy for presumed Graves disease and is now on thyroid hormone replacement. An older brother of the propositus also was noted to have low body weight. The father and a younger brother were unaffected. Thyroid function tests (TFTs) of the propositus and his older brother were abnormal, with high total T4 of 16.9 and 21.9 [mu]g/dl (5-11.6) respectively, high T3 of 381 and 580 ng/dl (80-200) and high FT4 index of 25.4 and 30 (6-10.5) together with undetectable TSH. In addition, the two affected boys had elevated 24-hour radioiodide uptake of 67 % and 90%, respectively (normal 15-25%) and mild thyroid enlargement on ultrasound.[br]Sequencing the [italic] TSHR [/italic] gene of the propositus revealed a heterozygous mutation in codon 431, GGC to AGC, resulting in the substitution of the normal glycine with serine G431S, in the first transmembrane domain. His affected brother harbored the same mutation, which was inherited from the mother, thus indicating that she was previously misdiagnosed with Graves disease. The two affected boys are currently being treated with methimazole, with the plan to undergo radioiodine ablation.[br]This mutation was previously reported in a family with nonautoimmune hyperthyroidism (1). Functional studies showed that expression of the mutant TSHR resulted in a greater cAMP accumulation and greater inositol phosphate formation compared with the normal TSHR, and that the mutant TSHR had increased TSH binding compared to the normal TSHR.[br]Conclusion: We report 2 boys with familial nonautoimmune hyperthyroidism due to a heterozygous gain of function mutation in the [italic] TSHR [/italic] gene inherited from their mother, who was previously misdiagnosed with Graves disease and underwent thyroid ablation. Even though nonautoimmune hyperthyroidism is a very rare condition, it should be considered in the differential diagnosis when evidence for autoimmunity is lacking. Genetic confirmation is required as it has important implications in the choice of treatment and for diagnosis of other family members.[br][br](1) Beibermann H at al., JCEM 2001; 86:4429.[br][br]Sources of Research Support: NIH Grant DK15070 awarded to SR, and DK091016 awarded to AMD. University of Southern Denmark medical student grant awarded to CL.[br][br]Nothing to Disclose: CL, LPK, REW, AMD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2017 308 2283 MON-393 PO56-03 Monday 1952 2012


1948 ENDO12L_MON-394 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Transient Neonatal Hypothyroidism in a Boy with Unbalanced Translocation t(8;16) Luciana Antunes de Almeida Secchi, Juliana Forte Mazzeu, Mara Santos Cordoba, Iris Ferrari, Helton Estrela Ramos, Francisco de Assis Rocha Neves University of Bras[iacute]lia, Bras[iacute]lia, Brazil; University of Bras[iacute]lia, Bras[iacute]lia, Brazil; University of Bras[iacute]lia, Bras[iacute]lia, Brazil; Federal University of Bahia, Salvador, Brazil Background: One of the remaining challenges in thyroid diseases physiopathology is etiologic diagnosis of congenital or neonatal hypothyroidism. Unlike permanent congenital hypothyroidism, in which about 15% of the patients have defects in one of the various genes involved in hormone synthesis, identification of genetic abnormalities causing the transient form of the disease is extremely rare.[br]Clinical case: A boy born after an uneventful full term pregnancy had ocular hypertelorism, deviated nasal septum, choanal atresia, umbilical hernia, hypospadia, imperforate anus and bradycardia. Neonatal screening test for congenital hypothyroidism was negative (TSH 7.09 [mu]U/ml, cut-off level 20 [mu]U/ml; total T4 16.4 [mu]g/dl, reference range 4.5-22.20 [mu]g/dl). In the third month of life severe hypothyroidism was diagnosed (serum free T4 undetectable; serum TSH 4.97 [mu]U/ml, reference range 0.49-4.67 [mu]U/ml). Levothyroxine reposition was then initiated and six weeks later thyroid function tests were normal. At the chronological age of seven months his bone age was compatible with that of a one-month-old baby, what corroborates the diagnosis of congenital hypothyroidism. When the boy reached the age of four, thyroid hormone replacement therapy was discontinued. Now aged 11, he remains euthyroid and growing steadily, without any signs of puberty. He has mental retardation, dysarthria and impaired walking. Chromosomal analysis after G-banding defined the patient[apos]s karyotype as 46,XY,der(8)t(8;16)(q24.3;q22)mat,16qh+. Array-CGH with patient[apos]s DNA revealed a [sim]80 kb terminal deletion on chromosome 8q24.3qter and a [sim]21 Mb duplication on chromosome 16q22qter. A balanced translocation was found in other three phenotypically normal family members (the patient[apos]s mother, one maternal uncle and the maternal grandfather). One deceased maternal uncle had phenotypic similarities with the patient, including hypothyroidism.[br]Conclusions: Only chromosomal imbalances involving DUOX/DUOXA genes, mapped to chromosome 15, have previously been described in association with transient neonatal hypothyroidism. So, this is the first report of association of a chromosomal imbalance involving chromosomes other than chromosome 15 and the transient form of the disease. This description contributes to the definition of the phenotype of the unbalanced translocation t(8;16) that has never been described before and opens new avenues for understanding the physiopathology of transient congenital hypothyroidism.[br][br]Nothing to Disclose: LAdAS, JFM, MSC, IF, HER, FdARN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 772 308 2284 MON-394 PO56-03 Monday 1953 2012


1949 ENDO12L_MON-395 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Familial Dysalbuminemic Hyperthyroxinemia with a Coexisting TSH-Secreting Pituitary Adenoma Marisabel Bravo, Rafael Arrojo E Drigo, Violet Lagari-Libhaber, Antonio Bianco, Brian Won-Sik Kim, Atil Kargi University of Miami Miller School of Medicine, Miami, FL Background: Patients with familial dysalbuminemic hyperthyroxinemia (FDH) are described as having elevated plasma concentrations of total T4 (T4) and free T4 (FT4) measured by chemiluminescence immunoassay (CLIA) and normal TSH and T3, whereas FT4 measured by direct dialysis is normal. Clinical Case: A 51 year old Hispanic woman was evaluated for elevated TSH ranging between 5.1-6.2 uIU/ml (reference 0.49-4.67). While her T4 was above the measurable range, and FT4 by CLIA was increased to 3-4.9 ng/dl (reference range 0.71-1.85), her FT4 by direct dialysis was normal at 1.9 ng/dl (reference 0.8-2.7). Record review revealed that five years prior to presentation, the TSH and FT4 (CLIA) had been normal, whereas the T4 was already elevated. Thyroxine binding protein assay demonstrated a marked increase in T4 binding to albumin consistent with FDH. Strikingly, thyroid function tests and binding assay in the patient[apos]s sister were also consistent with FDH. Workup of the rising TSH/T4/FT4(CLIA) in the setting of FDH included TPO antibodies (52.3 IU/ml, 5-34) and heterophile antibodies (negative). MRI of the sella revealed a pituitary macroadenoma measuring 2.4x3.4x2.2cm invading the right cavernous sinus. To examine the possibility of a TSH secreting pituitary adenoma versus FDH with superimposed Hashimoto[apos]s thyroiditis, a short T3 suppression test was performed with liothyronine 100mcg daily. This led to normalization but not suppression of TSH to 1.74 uIU/ml on the 8th day of treatment. Interestingly, FT4 remained elevated at 3.1 ng/dl. Alpha subunit and the calculated alpha:TSH molar ratio were elevated. Since the tumor was deemed inoperable, a trial of octreotide LAR was initiated during which TSH decreased from [sim]6.0 uIU/ml to 3.9 uIU/ml, FT4 assay decreased from [sim]4 ng/dl to 2.5 ng/dl, and T4 decreased from [gt]25 mcg/dl to 19 mcg/dl (reference range 4.5-12). However, the FT4 by direct dialysis was 2.3 ng/dl, slightly higher than prior to treatment. After 5 months of treatment, the tumor decreased from 3.4cm to 2.7cm suggestive of TSH secreting pituitary adenoma. Treatment was self-discontinued for 4 months after which TSH increased to 9.86 uIU/ml along with the FT4 (CLIA) 4.6 ng/dl, T4 [gt]25 mcg/dl, and T3 2.69 ng/ml (reference 0.79-1.49). FT4 by direct dialysis was now elevated at 4.2ng/dl, consistent with TSH secreting pituitary adenoma. Conclusion: This is the first reported case of FDH with a coexisting TSH-secreting pituitary adenoma.[br][br]Nothing to Disclose: MB, RAED, VL-L, AB, BW-SK, AK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1676 308 2285 MON-395 PO56-03 Monday 1954 2012


1950 ENDO12L_MON-396 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Thyroid Hormone Resistance: A Clinical Rather Than Genetic Diagnosis? Deepashree Gupta, Uzma Khan University of Missouri-Columbia, Columbia, MO [bold]Background[/bold]: To describe clinical presentation of thyroid hormone resistance and to emphasize that the currently available mutation analysis may not screen for all causative mutations necessitating a high index for clinical suspicion.[br][bold]Clinical Case[/bold]: 40 year old Caucasian male presented with a history of palpitations of 10 years duration. PCP started levothyroxine for high TSH but patient developed worsening palpitations and tremor. He was referred to Endocrinology and was noted to be clinically hyperthyroid. TSH: 12.58 (0.34-4.9), FT4: 4.18 (0.59 - 1.17) and FT3: 26.0 (2.0 -4.4). RAIU and scan: 6 h uptake- 74 % and 24 h uptake- 72 %. Uptake was uniform with enlargement of thyroid gland - right lobe more than left. He was diagnosed with TSH dependent hyperthyroidism. Pituitary MRI was negative for an adenoma. TPO and TSI antibodies were negative - [lt]0.3 and 89% respectively. Review of old labs showed mildly elevated TSH (5-7 range) without associated FT4.[br]Levothyroxine was discontinued and methimazole and propranolol started. Patient reported significant symptomatic improvement but TSH increased to 58.6 from 12.5 at initial clinic visit 5 months ago and thyroid gland began rapidly enlarging causing compressive symptoms. Thyroid gland enlargement was confirmed by thyroid ultrasound. At first clinic visit, thyroid ultrasound showed right lobe 5.4 x 2.2 x 2.8 cm, left lobe 4.9 x 1.8 x 1.8 cm, isthmus 14 mm, mildly increased vascularity, no discrete nodules. 5 months later the right lobe measured 7.3 x 3.1 x 3.7 cm, left lobe measured 6.1 x 2.3 x 2.6 cm, isthmus 17 mm with increased blood flow. Thyroid FNA and flow cytometry were benign. Mutation analysis for Resistance to Thyroid Hormone (RTH mutation by Quest) was negative. CT scan neck, chest and abdomen showed enlarged thyroid ([sim] 7 cm) but no evidence of pancreatic or lung mass.[br]Patient was referred to thyroid hormone resistance specialist and methimazole gradually tapered off. He and his family members are currently being screened for a novel mutation causing thyroid hormone resistance.[br][bold]Conclusion[/bold]: Thyroid hormone resistance is a group of rare disorders characterized by reduced end-organ responsiveness to thyroid hormone (TH). Symptoms usually worsen if an attempt to normalize TH is made either by radio-active iodine ablation or anti-thyroid medications. Commercial testing does not screen for all mutations and if diagnosis is suspected clinically, further testing is required to assess for novel mutations.[br][br]Nothing to Disclose: DG, UK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1254 308 2286 MON-396 PO56-03 Monday 1955 2012


1951 ENDO12L_MON-397 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Liothyronine Titration in the Treatment of Goiter in Thyroid Hormone Resistance Delia Viisoreanu, Monica Rodriguez Virginia Mason Medical Center, Seattle, WA Background: Every other day administration of liothyronine has been shown to be effective for the treatment of goiter in thyroid hormone resistance. Previously, suppression of TSH to 0.1 mU/L or less was needed prior to significant decrease in goiter size (1). This required liothyronine doses of 250 mcg every other day.[br]Clinical case: A 24 year old woman presented with symptoms of hyperthyroidism consisting of tachycardia, hand tremor, and goiter. She was found to have elevated free T4 at 2.3 ng/dL (0.7 - 1.9 ng/dL) and elevated free T3 at 6.7 pg/mL (2.3 - 4.2 pg/mL) with high thyroglobulin at 760 ng/mL (2 - 35 ng/mL), and inappropriately normal TSH at 1.6 mU/L (0.5 - 4.7 mU/L). MRI of the pituitary was normal. She had normal ratio of alpha subunit to whole TSH, which made TSH-producing pituitary adenoma unlikely. She also had negative thyroid stimulating immunoglobulins. She was diagnosed with thyroid hormone resistance. Her entire thyroid receptor beta gene was sequenced and no mutations were found. Therefore, it is possible that her thyroid hormone resistance is caused by a mutation in one of the cofactors that interact with the thyroid hormone receptor, as described previously (2). Because thyroid hormone resistance is associated with a higher risk of autoimmune thyroid disease, her anti-thyroglobulin and anti-thyroid peroxidase antibodies were measured, but were found to be undetectable.[br]She was treated with atenolol 50 mg daily for tachycardia, palpitations, and tremor with improvement in her symptoms. For her goiter, liothyronine was started at 25 mcg every other day, and was uptitrated. She had significant decrease in her goiter with liothyronine 175 mcg every other day, which resulted in reduction of her TSH to 0.6 mU/L. [br]Conclusion: This case shows that lower doses of liothyronine than previously reported are already effective in the treatment of goiter in thyroid hormone resistance. This occurred in the setting of TSH suppression to only 0.6 mU/L in our patient. We suggest that liothyronine should be titrated to effect regardless of TSH level because this may result in a lower liothyronine dose and thus reduce the risk of thyrotoxicosis.[br][br](1) Refetoff S, Anselmo J. Regression of a large goiter in a patient with resistance to thyroid hormone by every other day treatment with triiodothyronine. Thyroid 2004;14:71-74. (2) Weiss RE, Hayashi Y, Nagaya T, Petty KJ, Murata Y, Tunca H, Seo H, Refetoff S. Dominant inheritance of resistance to thyroid hormone not linked to defects in the thyroid hormone receptor alpha or beta genes may be due to a defective cofactor. J Clin Endocrinol Metab 1996;81:4196-4203.[br][br]Nothing to Disclose: DV, MR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 790 308 2287 MON-397 PO56-03 Monday 1956 2012


1952 ENDO12L_MON-398 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Diagnosing Resistance to Thyroid Hormone in a Patient with History of Graves Disease Sravanthi Nagavalli, Anthony Yin, Silvana Marisa Giannelli, Andrew George Gianoukakis Harbor - UCLA Medical Center, Torrance, CA [bold]Background[/bold]: Resistance to thyroid hormone (RTH) is characterized by elevated thyroid hormone (TH) levels with a normal or slightly elevated TSH suggestive of target tissue hypo responsiveness to TH. RTH can coexist with Graves[apos] disease (GD) which represents a diagnostic challenge as illustrated by the case below.[br][bold]Clinical Case[/bold]: Thirty year old Korean female presented with palpitations. On evaluation, patient had a goiter and tachycardia. FT4 = [gt]6 ng/dl (n: 0.73- 1.95 ng/dl), T3= 749 ng/dl (n: 79-149 ng/dl) and TSH=0.01 [micro]IU/dl (n: 0.34-5.6 [micro]IU/dl). I123 uptake was uniform, 82% at 2 hours, and 81% at 24 hours. TSI were not ordered. Therapy with Methimazole (MMI) was initiated with a presumptive diagnosis of hyperthyroidism due to GD. After 7 weeks, TSH=0.04 [micro]IU/dl, T3=344 ng/dl and FT4=2 ng/dl. Radioactive iodine (RAI) ablation with 29 mCi I131 was performed. MMI was resumed post ablation. Four months after ablation, MMI was discontinued as TSH=8.7 [micro]IU/dl, T3=126 ng/dl and FT4= 1.1 ng/dl. One month later the TSH normalized to 1.55 [micro]IU/dl, T3 and FT4 were high at 164 ng/dl and 2.93 ng/dl respectively. Over the next 3 years TSH continued in the normal to mildly elevated range (1.5 to 7.46 [micro]IU/dl) while T3, FT4 and Total T4 (TT4, n: 6.1-12.2 mcg/dl) remained slightly elevated at 130-182 ng/dl, 1.85-2.93 ng/dl and 14.4-17.6 mcg/dl respectively. The patient was asymptomatic and not treated with levothyroxine. Alternative diagnoses including RTH, TSH secreting pituitary tumor, excess TBG and familial dysalbuminemic hyperthyroxinemia (FDH) were considered. MRI brain showed an incidental pars media cyst. T3 suppression testing resulted in a suppression of TSH from 3.24 to 0.13 [micro]IU/dl and decrease in FT4 from 2.2 to 1.3 ng/dl. The [alpha] subunit was 0.6ng/ml (n [lt] 1.5 ng/ml) with corresponding TSH=2.9 [micro]IU/dl and TT4=16.1. SHBG=13 nmol/L (n: 14-102).Thyroid binding globulin=22.4 mcg/ml (n: 13.5-30.9 mcg/ml). FT4 by equilibrium dialysis=3.6 ng/dl (n: 0.8-2.7 ng/dl). TSH secreting tumor and binding protein abnormalities were ruled out. RTH was highly suspected. THR[beta] gene sequencing revealed a novel mutation S350L. Thus, the diagnosis of RTH was made in this patient after RAI ablation for GD.[br][bold]Conclusion[/bold]: RTH and GD can coexist. Biochemical evidence suggestive of RTH may not be apparent until the [quot]confounding effects[quot] of GD hyperthyroidism are eliminated. TSH secreting pituitary tumor, excess TBG and FDH must be considered in the differential diagnosis and excluded.[br][br](1)Weiss R, Refetoff S. Resistance to Thyroid Hormone. Endocrine [amp] Metabolic Disorders 2000; 1:97-108.[br][br]Nothing to Disclose: SN, AY, SMG, AGG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 705 308 2288 MON-398 PO56-03 Monday 1957 2012


1953 ENDO12L_MON-399 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Resistance to Thyroid Hormone [ndash] A Novel Mutation Bonnie Goldsmith, Daniel Hsia, Lefkothea Karaviti Baylor College of Medicine, Houston, TX Introduction: Resistance to thyroid hormone (RTH) is a syndrome of reduced sensitivity to thyroid hormone (TR). In the majority or cases it is caused by mutations of the thyroid hormone receptor beta ([italic]TR[beta][/italic]) gene.[br]Background: Resistance to thyroid hormone was first described by Refetoff, DeWind, and DeGroot in 1967.[br]Context: We report the case of a 5 year old female with thyromegaly, high serum T4 and T3 concentrations with non-suppressed TSH, consistent with RTH. There is no family history of thyroid problems or goiter.[br]Clinical Course: The patient was brought to medical attention at 3 years of age because of hyperactivity prompting thyroid testing. On presentation to our clinic, she had mild thyromegaly. Thyroid function test results were: T3 214 ng/dl (90-260 ng/dl), FT4 by dialysis 4.6 ng/dl (1.0-2.4 ng/dl) and TSH 2.3 mIU/L (0.32-5.0 mIU/L) with negative thyroid autoantibodies. Iodine uptake was 33% at 24 hours (10-25%). Based on the elevated T4 with non-suppressed TSH, we suspected RTH.[br]Outcomes: Given the above results, we sequenced the [italic]TR[beta][/italic] gene. A missense mutation was identified. It consists of a G to T transversion (c.793) in exon 8, which predicts the replacement of aspartate (GAC) by tyrosine (TAC) at codon.265 (D265Y). This novel mutation is predicted to be damaging according to the polyphen model. The mutation is located at the proximal end of the ligand binding domain and involves the mutagenic cluster 3. Thus, the mutation most likely affects T3-binding to the TR[beta] molecule. Inability to bind T3 can cause dysregulation of the pituitary thyroid axis as well as reduced sensitivity to TH at the peripheral tissues level.[br]Conclusions: We have identified a novel mutation in the TR[beta] gene which is expected to be responsible for the clinical phenotype of RTH. Further studies are underway to evaluate the structure and function of the receptor in light of this mutation. In addition, studies will be undertaken to determine whether this mutation affects other members of the family or occurred de novo.[br][br]Nothing to Disclose: BG, DH, LK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1815 308 2289 MON-399 PO56-03 Monday 1958 2012


1954 ENDO12L_MON-400 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Noncompliance? Not Necessarily. A Case of Sevelamer Inhibition of Levothyroxine Absorption Christopher Wong, Kevin Codorniz, Sergio Infante Loma Linda University, Loma Linda, CA; Loma Linda University, Loma Linda, CA; Loma Linda University, Loma Linda, CA [bold]Background[/bold][br]The link between thyroid hormone and renal function has been well documented, as hypothyroidism is known to blunt kidney size in congenital cases, leading to a higher incidence of congenital renal abnormalities. Elevated TSH has also been associated lower eGFR and a higher rate of CKD.[br][bold]Purpose[/bold][br]Hypothyroidism is common among patients with end stage renal disease (ESRD), and management of both conditions can be difficult for both physicians and patients. Many medical conditions, drugs, and even foods can impair absorption of Levothyroxine, leading to refractory hypothyroidism. It is important to investigate all causes of refractory disease before attributing it to medication non-compliance. We present a case of phosphate binder inhibition of Levothyroxine absorption and subsequent improvement in symptoms after administration of levothyroxine sublingually.[br][bold]Case[/bold][br]Patient is a 30 YO Hispanic male with a history of ESRD secondary to obstructive uropathy, on HD since age 18, with thyroidectomy and parathyroidectomy in 2006 due to a parathyroid adenoma. Patient had several admissions for pericardial and pleural effusions. Among other medications, he was taking sevelamer 4000mg TID and liothyronine 0.05mg daily. He maintained that he had been compliant with his thyroid medication despite frequently elevated TSH, and persistent symptoms. On exam he presented with central obesity, loss of distal portion of eyebrows, and depressed mood. He was subsequently started on levothyroxine sublingually and avoided phosphate binders in the morning. Within one month, TSH levels had decreased substantially from the 600[apos]s to the 20[apos]s and free T4 was now in normal range. Symptoms also improved; including reduction of pericardial effusion and greater hemodynamic stability during HD. He also reported increased physical activity and improved mood.[br][bold]Discussion[/bold][br]Conditions that impair levothyroxine absorption include: chronic gastritis and celiac disease; as well as medications such as cholestyramine, ferrous sulfate, and sucralfate. Research has shown that phosphate binders including sevelamer decrease intestinal bioavalibility of levothyroxine. The amount of time in years using phosphate binders is correlated with increasing TSH levels. It is possible that absorption can be improved by taking phosphate binders and levothyroxine at different times in the day, or using calcium acetate if possible since it has been shown to affect absorption less.[br][br]Nothing to Disclose: CW, KC, SI 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1442 308 2290 MON-400 PO56-03 Monday 1959 2012


1955 ENDO12L_MON-401 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) Supervised Weekly Dosing of Levothyroxine To Demonstrate and Circumvent Pseudomalabsorption Chaitanya Kumar Mamillapalli, Deepika Nallala, Michael Jakoby Southern Illinois University School of Medicine, Springfield, IL In healthy and non-pregnant adults with hypothyroidism, a thyroxine dose of approximately 1.6 [mu]g/kg/d is usually adequate to ameliorate symptoms and restore biochemical euthyroidism. While dose requirements may be increased by factors that limit absorption (reduced gastric acidity), increase excretion (nephrotic syndrome), or accelerate metabolism (e.g. carbamazepine) of thyroxine, medical non-compliance is the most common etiology of treatment failure. We report a case in which pseudomalabsorption was clearly demonstrated by directly observed, weekly administration of thyroxine in the office.[br]A 25 year-old woman was referred for evaluation of persistent clinical and biochemical hypothyroidism despite prescription of very high doses of thyroxine and liothyronine. Thyroidectomy had been performed to manage Graves[apos] thyrotoxicosis and goiter with compressive symptoms. Despite reported compliance, the patient experienced several months of persistently elevated TSH, undetectable T4, and symptoms of hypothyroidism such as fatigue and constipation. Evaluation for celiac sprue was unrevealing. The patient was educated to take thyroxine with water and on an empty stomach remote from food and mineral supplements. When confronted with the possibility of non-compliance, she changed primary care providers. At time of consultation, she was reportedly taking Levoxyl[trade] brand thyroxine 600 [mu]g daily and Cytomel[trade] brand liothyronine 25 [mu]g daily.[br]Examination was notable for a well-healed low neck incision and deep tendon reflex delay. TSH was 133 mIU/L (ref. range 0.40-4.00), and total T4 was undetectable. The patient[apos]s pharmacy confirmed that she had not been filling her prescriptions. Weekly administration of 616 [mu]g of Synthroid[trade] brand thyroxine (seven 88 [mu]g pills) in the office was offered as alternative therapy, and the patient agreed. After 4 weeks, TSH fell to 4.74 mIU/L, and she acknowledged improvement of hypothyroidism-related symptoms.[br]Weekly and directly observed thyroxine therapy confirmed medical non-compliance as the etiology of persistent hypothyroidism in our patient[apos]s case and resulted in significant biochemical and clinical improvements. In direct comparisons, the biochemical and clinical effects of daily or weekly administration of equivalent thyroxine doses are indistinguishable. Supervised weekly administration of thyroxine should be considered to confirm pseudomalabsorption and circumvent non-compliance in treatment of hypothyroidism.[br][br]Nothing to Disclose: CKM, DN, MJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1095 308 2291 MON-401 PO56-03 Monday 1960 2012


1956 ENDO12L_MON-402 POSTER SESSION: Thyroid Disease Unusual Case Reports (1:30 PM-3:30 PM) A Case of L-T4 Malabsorption Caused by a Proton Pump Inhibitor and Solved by a Novel Formulation of L-T4 Roberto Vita, Salvatore Benvenga University of Messina, School of Medicine, Messina, Italy; University of Messina, School of Medicine, Messina, Italy; University Hospital, AOU Policlinico G Martino, Messina, Italy Interference of nonabsorbable and absorbable anti-ulcer drugs on the intestinal absorption of LT4 has been reported. Interference of PPI on LT4 absorption is attributable to poor solubilization of LT4 in nonacid pH. The typical approach is to increase the daily dose of L-T4, but this would expose the patient to the risk of iatrogenic hyperthyroidism if the patient decreases or quits the PPI. In an in vitro test, a new formulation of LT4 in soft gel capsules (Tirosint[reg] capsule, IBSA, Switzerland) was much more consistent in releasing the hormone regardless of changes in the pH compared to either brand or generic L-T4 tablet (1).[br]A woman with Hashimoto[apos]s thyroiditis-associated subclinical hypothyroidism (serum TSH [gt] 6.8 mU/L) had been treated, by others, with brand LT4 tablet (100 mcg/d). Because of failure of TSH to become fully normal ([ge] 4.0 mU/L), the daily dose had been progressively increased to 125 and 150 mcg/d, with serum TSH levels of 2.4 and 0.6 mU/L. She was referred to us for investigating possible malabsorption of LT4. At history, the lady disclosed gastroesophageal reflux, for which she used to take a PPI (pantoprazol) almost daily. No other drugs were taken. We switched the LT4 150 mcg/d tablet to 125 mcg/d capsule for 2 months and 100 mcg/d for another 2 months, while maintaining pantoprazol. Serum TSH was lower than under the equivalent regimens with the tablet: 0.46 (vs. 2.4) and 2.4 (vs. [gt]4.0 mU/L). Upon switching back to the tablet (100 mcg/d) while maintaining pantoprazol, serum TSH bounced to 3.2 mU/L two months after this switch. We also evaluated the intestinal absorption of LT4 acutely, i.e. by administering 600 mcg LT4 in tablet or capsule with 240 mL water. In either occasion, we measured serum T4 at baseline and various times up to 4 h, and then corrected the postdose serum levels for baseline. The two curves of serum T4 showed better absorption indices for the capsule vs. tablet (AUC0-4h = 16240 vs. 12710 [+27.8%]), with a maximum absorption (Cmax) of 108 vs. 73, and time of maximum absorption (Tmax) at 120 min vs. 180 min.[br]This case demonstrates the practical application expected on the said in vitro data, namely an anticipated good absorption of LT4 in soft gel capsules because dissolution (and therefore availability at the intestinal mucosa) is negligibly affected by changes in the ambient pH.[br][br](1) Pabla D et al, Eur J Pharm Biopharm. 2009; 72:105.[br][br]Nothing to Disclose: RV, SB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1106 308 2292 MON-402 PO56-03 Monday 1961 2012


1957 ENDO12L_MON-403 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Analytical Performance and Clinical Utility of a TSH Receptor Bioassay for Thyroid-Stimulating Autoantibodies George J Kahaly, Tanja Diana, Johannes C Leschik, Yunsheng Li, Paul D Olivo Gutenberg University Medical Center, Mainz, Germany; Diagnostic Hybrids, Inc, Athens, OH [bold]Background and aims: [/bold]A bioassay based on a chimeric TSH receptor (TSHR) for measurement of thyroid stimulating autoantibodies (TSAb) has been recently introduced. Here we compared the analytical performance of this bioassay with an automated electrochemiluminescence TSHR binding inhibiting immunoglobulin (TBII) assay and evaluated its clinical utility using dilution studies.[br][bold]Methods: [/bold]Limit of Detection (LoD) and Quantitation (LoQ) were measured using a stimulating human monoclonal antibody (M22). Serial dilutions were performed on 237 serum samples of 40 hyperthyroid patients with Graves[apos] disease prior to and during antithyroid drug treatment (ATD). The titer was defined as the first dilution step below the cut-off for each assay.[br][bold]Results: [/bold]LoB was 77% (percentage specimen to reference ratio) and 0.5 IU/L for the TSAb bioassay and the TBII assay, respectively. LoD and LoQ of the bioassay were 156 fold (0.02 ng/ml vs. 3.125 ng/ml) and 300 fold lower (0.02 ng/ml vs. 6 ng/ml) than for the TBII assay. Half maximal effective concentration of M22 in the bioassay (0.20 ng/ml) was 813-fold lower than in the TBII assay (162.7 ng/ml). In dilution studies there was a 21.8%, 41.8%, 22.8% and 13.8% difference of positive samples between the TSAb and TBII assays at dilutions 1:3, 1:9, 1:27 and 1:81, respectively (P[lt]0.003). The higher the dilution step, the higher was the positive TSAb/TBII ratio. In a subset of 85 samples with titer [ge]4, the TBII assay showed a dilution pattern with more rapid signal reduction than the TSAb assay. In 19 of 40 (47.5%) patients, TSAb titers were more informative than undiluted TSAb values. TSAb titers at all time points were 1.2 dilution steps higher than TBII titers (P[lt]0.001). A linear decrease of the TSAb titer was noted in the responders to ATD (euthyroidism); in contrast an increase was observed in the non-responders (persistent hyperthyroidism or relapse after dropping ATD). Baseline mean TSAb titers were 4.0[plusmn]1.8 vs. 2.7[plusmn]1.3 (P=0.027) and mean TBII titers were 2.9[plusmn]1.3 vs. 1.7[plusmn]0.8 (P=0.006) in the non-responders and responders to ATD, respectively. At week 12 of ATD therapy and thereafter, all differences of titers between responders and non-responders were highly significant (P[lt]0.001) for TSAb and TBII.[br][bold]Conclusions: [/bold]The chimeric TSHR bioassay has a higher analytical performance than the binding assay. Serial serum dilution analysis was clinically useful and prognostically informative in as many as 50% of patients with Graves[apos] disease.[br][br]Sources of Research Support: Research funding from Diagnostic Hybrids, Inc., Athens, OH, USA.[br][br]Disclosures: GJK: Research Funding, Diagnostic Hybrids, Inc. YL: Employee, Diagnostic Hybrids, Inc. PDO: Employee, Diagnostic Hybrids, Inc. Nothing to Disclose: TD, JCL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 690 309 2293 MON-403 PO40-01 Monday 1962 2012


1958 ENDO12L_MON-404 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Circulating IgGs May Modulate IGF-I Receptor Activity in a Subset of Euthyroid Patients with Graves Ophthalmopathy Aimee J Varewijck, Anita Boelen, Clementine JJ van Zeijl, Steven WJ Lamberts, Wilmar M Wiersinga, Leo J Hofland, Joseph AMJL Janssen Erasmus Medical Center, Rotterdam, Netherlands; Academic Medical Center, Amsterdam, Netherlands [italic]Context [/italic]There is a close relationship between TSH binding inhibitory immunoglobulins (TBII) and the activity of Graves Ophtalmopathy (GO). The IGF-I receptor (IGF-IR) has been proposed to be the second major autoantigen that might play a role in the causal relationship between autoimmunity and GO. In addition, it has previously been suggested that that a subset of Graves[apos] IgG contains antibodies that stimulate the IGF-IR.[br][italic]Objective[/italic] To study the relationship between TBII and circulating IGF-I bioactivity in euthyroid patients with GO.[br][italic]Methods [/italic]Cross-sectional study of 62 euthyroid Graves[apos] patients (TSH [lt]4.0 mU/L and free T4 [lt]23.0 pmol/L), who were referred to our combined thyroid[ndash] eye clinic for GO (m: 8, f: 54; age: 49.6 [plusmn] 1.8 yrs). IGF-I bioactivity was determined by the IGF-I KIRA assay and total IGF-I by immunoassay in serum samples. TBII was measured by immunoassay (Trak). For IgG depletion of serum, protein G magnetic beads were used.[br][italic]Results[/italic] Geometric mean TBII was 4.3 U/L (range: 0.9-163.0). IGF-I bioactivity was low-normal (mean: 248.1[plusmn]12.7 pmol/L; Z-score: -1.6[plusmn]0.1 SD), while total IGF-I was normal (mean: 23.5[plusmn]1.5 nmol/L; Z-score: 0.6[plusmn]0.2 SD).[br]Overall, there was no relationship between total IGF-I and age (r=-0.18, p=0.17), nor between IGF-I bioactivity and age (r=0.084, p=0.52). There was a significant inverse relationship between TBII and total IGF-I (r= -0.26, p=0.05), but not for IGF-I bioactivity (r=-0.17, p= 0.19).[br]In the highest tertile of TBII, we found a strong positive relationship between IGF-I bioactivity and age (r=0.56, p=0.009) and between TBII and T3 (r=0.56, p=0.01), while such relationships were absent in the lowest two tertiles. Moreover, in the highest tertile, there was no relationship between total IGF-I and age (r=0.16, p=0.48).[br]Depletion of IgGs from serum of 5 GO patients with the highest TBII resulted in a decrease in IGF-I bioactivity in 2 patients whereas it increased in 2 patients. In one patient, and in 2 healthy controls, IgG depletion did not affect IGF-I bioactivity.[br][italic]Conclusions[/italic] The strong positive correlations between IGF-I bioactivity and age and between TBII and T3 in the highest tertile of TBII, point towards stimulating activity of these immunoglobulins. This suggests that in a subset of euthyroid patients with GO circulating IgGs modulate IGF-IR activity.[br][br]Sources of Research Support: J.A.M.J.L. Janssen has received an unrestricted grant from Novo Nordisk A/S (Alphen aan de Rijn, the Netherlands).[br][br]Nothing to Disclose: AJV, AB, CJJvZ, SWJL, WMW, LJH, JAMJLJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1510 309 2294 MON-404 PO40-01 Monday 1963 2012


1959 ENDO12L_MON-405 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Long-Term Effects of High-Dose Methylprednisolone Pulse Therapy on Thyroid-Associated Ophthalmopathy Xiaomei Liu, Bingyin Shi, Li Qin, Shu Wang First Affiliated Hospital of Xi[apos]an Jiaotong University School of Medicine, Xi[apos]an, China; First Affiliated Hospital of Xi[apos]an Jiaotong University School of Medicine, Xi[apos]an, China Objective To report the long-term effects and safety of high-dose intravenous methylprednisolone pulse(IVMP)therapy on Thyroid Associated Ophthalmopathy (TAO). Design Prospective clinical trial. Methods 52 TAO patients were treated with high-dose IVMP therapy alone between Oct.2006 and Nov.2010, and their clinical manifestation and indices before, during and after therapy were recorded in details, with a 12-57months (mean 28.3months) follow-up. Results The cure-improve rate is 96.77% 1 month after treatment on ophthalmodynia, 89.59% on photophobia, 89.13% on lacrimation, 73.33% on eyelid swelling, 43.25% on diplopia, and the cure-improve rate at end of the follow-up is 90.33% on ophthalmodynia, 85.42% on photophobia, 82.61% on lacrimation, 80.00% on eye swelling, 83.79% on diplopia. Meanwhile, the visual acuity, eyeball activity, proptosis as well as the CAS was significantly improved after therapy, only two patients who associated with other clinical conditions suffered from liver injury during treatment or observation, and 4 patients had put on weight slightly. Conclusions High-dose IVMP therapy is a very effective, safe and well-tolerated treatment for TAO and associated with nearly no side-effects.[br][br]Nothing to Disclose: XL, BS, LQ, SW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 344 309 2295 MON-405 PO40-01 Monday 1964 2012


1960 ENDO12L_MON-406 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Plasma Exchange in the Pre-Operative Management of Severe Thyrotoxicosis Yavuz Yavuz, Nilubol Naris, Leitman F Susan, Celi S Francesco NIDDK-National Institutes of Health, Bethesda, MD; NCI-National Institutes of Health, Bethesda, MD; Clinical Center-National Institutes of Health, Bethesda, MD Untreated severe thyrotoxicosis carries a high risk of morbidity and mortality. In cases where medical treatment of hyperthyroidism in Graves[apos] disease is not tolerated, surgery is the recommended approach; however, preoperative optimization of thyrotoxicosis is mandatory to minimize the risk of intra- and post-operative complications. Plasma exchange is a short term treatment for thyrotoxicosis that reduces plasma protein-bound thyroid hormones and TSH receptor antibodies. We describe two patients with impending thyroid storm due to Graves[apos] disease who could not tolerate medical therapy. Plasma exchange was successfully used as a preparative therapy prior to thyroidectomy in both patients.[br]Case 1: A 38 y.o. female presented with weight loss, palpitations, irritability and an 80 gram goiter. She developed a severe skin reaction to methimazole and severe hepatotoxicity to propylthiouracil (PTU), with liver enzymes [gt] 4-fold upper normal limits. Thyroid stimulating immunoglobulin (TSI) titer was 4.8 (normal [ge]1.3 TSI index). PTU was discontinued and [sup]131[/sup]I 10 mCi was given, but 3 weeks later her clinical and biochemical status acutely worsened. She underwent 5 single-volume alternate-day plasma exchanges, resulting in clinical and biochemical improvement: fT4 decreased from [gt]8.0 to 3.2 ng/dL (n.r. 0.8-1.9) and T3 decreased from 1302 to 214 ng/dL (n.r. 90-215). Two weeks later, thyroid profile again worsened, with a Wartofsky-Burch score of 15. An additional exchange was performed, and she underwent thyroidectomy without complications.[br]Case 2: A 44 y.o. female with a 6-year history of Graves[apos] disease presented with worsening thyrotoxic symptoms, anemia, atrial fibrillation, and congestive heart failure. TSI titer was 6.0. She had not tolerated methimazole in the past due to intractable abdominal cramping, and on PTU continued to have atrial fibrillation with palpitations, exertional dyspnea, weakness, failure to thrive, and mild hepatotoxicity. Her Wartofsky-Burch score was 20. In preparation for thyroidectomy, she underwent two daily 1.5-volume plasma exchanges, with a decrease in FT4 from [gt]8.0 to 2.0 ng/dL and a decrease in T3 from 602 to 129 ng/dL. Postoperatively, her atrial fibrillation and cardiomyopathy resolved.[br]Plasma exchange is an effective, low-risk approach to rapidly optimize thyroid management prior to surgical thyroidectomy when thyroid storm is imminent and standard medical management is either ineffective or not tolerated.[br][br]Sources of Research Support: This work was supported by the Intramural Research Program of NIDDK-NIH.[br][br]Nothing to Disclose: YY, NN, LFS, CSF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1898 309 2296 MON-406 PO40-01 Monday 1965 2012


1961 ENDO12L_MON-407 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Association between Autoimmune Thyroid Diseases and Celiac Disease. The Tip of the Iceberg of the Autoimmune Polyendocrine Syndrome? Susana Mallea-Gil, Marta Aparicio, Karina Bertini, Carolina Ridruejo, Viviana Ercoli, Silvia Gimenez, Adriana Andrush, Vielka Cedeno-Arauz, Sandra Perez-Martinez, Noel Aldabe, Mariel Iriarte, Pablo Galarza, Adrian Perusco, Silvana Roveto, Estela Rivero, Carolina Ballarino Hospital Miltar Central, Buenos Aires, Argentina; Instituto de Investigaciones M[eacute]dicas Lanari, Buenos Aires, Argentina Patients with autoimmune thyroid diseases (AITDs) are likely to develop celiac disease (CD) among other autoimmune disorders. The frequency of this association in adults is about 1.2- 4.8%. CD symptoms are considerably different according to the onset age, and can vary from very severe to asymptomatic forms, the latter more frequent in adults.[br][bold]Objectives: [/bold]To determine the frequency of CD in adults patients with AITDs in one single medical center in Buenos Aires, Argentina, during the period January-December 2011. To assess the genetic profile and the presence of 21-hydroxylase antibody in the patients with AITDs-CD association.[br][bold]Material and method[/bold]: We studied a consecutive cohort of 242 patients. The following antibodies were screened: Tiroperoxidase by MEIA, TSH receptor by Radioreceptor assay, IgA Endomisium by IFI, IgA and IgG transglutaminase by ELISA and gliadin by ELISA. In 4 with positive CD, genetic studies: Class II HLA typification by PCR-SSOP and 21-hydroxylase by RIA were performed.[br][bold]Results[/bold]: In 242 patients with AITDs: 80% had Hashimoto[apos]s thyroiditis and 20% had Graves[apos] disease, 81.5 % were female, mean age: 50.8 years. Six patients were affected with previously diagnosed CD. Six out of 242 patients had positive CD serological testing. All of them (female) agreed to undergo endoscopy and duodenal biopsies. Histopathological signs of CD were found in 4 patients. Tiredness, alopecia, osteoporosis, infertility and anemia were found as predominant symptoms in our patients. Fourteen out of 248 AITDs patients had an autoimmune illness other than CD, none of them were positive for the AITDs-CD association. In 3/4 patients with this association, 21-hydroxylase antibody was positive, suggesting Autoimmune Polinedocrine Syndrome Type II (APS II) and 1 was negative (APS III). In patients with APS II, HLA DRB1* 04 was present in two of them, HLA DRB1*03:01 in one and HLA DRB1*08 in one, HLA DQB1*02:01 was present in two patients and HLA DQB1*03:02 in all of them. The patient with APS III was carrier of HLA DRB1*04, HLA DQB1*02:01 and HLA DQB1*03:02.[br][bold]Conclusions[/bold]: The frequency of the AITDs-CD association in our adult patients was 3.7%. CD symptoms were silent in our patients previously diagnosed with AITDs. We found no differences in HLA typing between patients with APS II and III, similar to our previous study in some Argentine families. Positive 21-hydroxylase antibody was present in some of these patients, so asymptomatic adrenal autoimmunity was detected.[br][br]Nothing to Disclose: SM-G, MA, KB, CR, VE, SG, AA, VC-A, SP-M, NA, MI, PG, AP, SR, ER , CB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 235 309 2297 MON-407 PO40-01 Monday 1966 2012


1962 ENDO12L_MON-408 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Radioiodine Therapy for Men with Graves Disease: Efficacy Analysis and Outcomes in Drug Na[iuml]ve Patients and Those on Antithyroid Drugs C Shivaprasad, KM Kumar Prasanna M S Ramaiah Medical College and Hospital, Bangalore, India; PRIMER, CDEC, HBR Layout, Bangalore, India [bold]Objective[/bold]: The objective of this retrospective study was to compare the efficacy of empirical doses of radioiodine between drug na[iuml]ve patients and patients who had previously received anti-thyroid drugs(ATD) in male patients with Graves[apos] disease(GD).[br][bold]Methods:[/bold] The clinical outcome of 335 male patients with GD treated with I131 from 1998-2008 was evaluated. 187 patients were drug na[iuml]ve and 148 had received ATD before I131. Mean duration of ATD therapy before I131 was 23 months. Patients were treated with a single empirical dose of radioiodine(RAI) based on the uptake of RAI and thyroid size. Following RAI treatment, thyroid status was monitored every 2-3 months during first year and 3-6 monthly thereafter. All the patients in study group were followed up for at least 12 months after treatment.[br][bold]Results:[/bold] The overall success rate of single dose of RAI was 80.2%(275/335). The cumulative rate of hypothyroidism at 1 year was 65%. In drug na[iuml]ve patients the mean dose of I131 was 8.57[plusmn]2.86 mCi. The overall success rate of single dose of RAI was 91.4%. The cumulative rate of hypothyroidism till the last follow up date was 78.1%. 13.3% of the patients remained euthyroid till the last follow-up. Median duration for hypothyroidism was 5 months (2-81 months) and mean duration was 10.23 months. Worsening of ophthalmopathy was seen in 6.9% of patients.Cumulative rates of hypothyroidism between 1-6 months and 6-12 months were 55.1% and 12.2% respectively. In patients who had received ATD, the mean dose of I131 was 8.7 [plusmn]2.98 mci. The overall success rate of single dose of RAI was 82.3%. The cumulative rate of hypothyroidism till the last follow up date was 69.7%. 12.8 % remained euthyroid till last follow-up. Median duration for hypothyroidism was 5 months(2-170 months) and mean duration was 13.42 months. Worsening of ophthalmopathy was seen in 9.5% of patients. Cumulative rates of hypothyroidism between 1-6 months and 6-12 months were 44.6% and 16.9% respectively.[br][bold]Conclusions:[/bold] Drug na[iuml]ve men with GD treated with I131 had higher remission rate and cumulative rate of hypothyroidism compared to patients who had previously received ATD. They became hypothyroid earlier and had higher cumulative rate of hypothyroidism between 1-6 months after i131 compared to the latter. Patients who had received ATD had higher rate of worsening of ophthalmopathy and higher cumulative rates of hypothyroidism between 6-12 months after I131 compared to drug na[iuml]ve patients.[br][br]Nothing to Disclose: CS, KMKP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 817 309 2298 MON-408 PO40-01 Monday 1967 2012


1963 ENDO12L_MON-409 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Siglec1 (Sialic Acid-Binding Immunoglobulin-Like Lectin1) as a Novel Predictive Marker for Recurrence of Graves Disease Koshi Hashimoto, Emi Ishida, Takuya Tomaru, Atsushi Ozawa, Nobuyuki Shibusawa, Tetsurou Satoh, Masanobu Yamada, Masatomo Mori Gunma University Graduate School of Medicine, Maebashi, Japan [Background] Therapy using anti-thyroid drugs against Graves[apos] disease is the most common and the first choice in Japan and the remission rate of is about 90%. On the other hand, the recurrence rate is also high, which is a weakness of the therapy. However, to date, there are no reliable tests to predict the recurrence or relapse of the disease. To search for a novel diagnostic marker to predict the recurrence of Graves[apos] disease, we performed DNA microarray and examined gene expression in leukocytes of both a frequently relapsing patient with Graves[apos] disease (R) and a patient in long-term remission after the cessation of medication (non-R). We found that Sialic acid-binding immunoglobulin-like lectin1 (Siglec1) gene expression was significantly increased in the R patient. To analyze whether Siglec1 may be a predictive marker for the recurrence of Graves[apos] disease, we evaluated Siglec1 mRNA levels in the leukocytes of Graves[apos] disease patients. [Patients and Methods] Seventy-seven patients diagnosed as having Graves[apos] disease at the thyroid clinic in Gunma University Hospital have been included in the study since 2006. RNA was extracted from total venous blood followed by reverse-transcription (RT). We evaluated Siglec1 mRNA levels by quantitative-RT-PCR using the Siglec1 specific Taqman[reg] probe. [Results] Siglec1 mRNA levels in the R patients were significantly higher than those in the non-R patients. Based on the ROC curve prepared with Siglec1 gene expression levels and the history of recurrence, we set 255 gene copies of Siglec1 as a cutoff value. Employing this cutoff value, we divided the patients into two groups; high and low-copy number groups and examined the history of recurrence. By the chi-square test, the Siglec1 gene copy number was significantly related to recurrence (p[lt]0.0001, sensitivity 82.6%, specificity 90.7%). However, no relationship between TSH receptor antibody titer (binding rate) and recurrence was observed in the current study. Since, Siglec1 gene expression levels are not related to serum TSH levels or thyroid hormone levels, the mRNA levels are not affected by patient[apos]s thyroid status. [Conclusion] Siglec1 gene expression levels in the leukocytes may be a novel predictive marker for recurrence.[br][br]Nothing to Disclose: KH, EI, TT, AO, NS, TS, MY, MM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 885 309 2299 MON-409 PO40-01 Monday 1968 2012


1964 ENDO12L_MON-410 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Short [ldquo]On[rdquo] Followed by Rapid [ldquo]Half-Cut[rdquo] of Methimazole (MMI) for Initial Treatment of Graves Disease (GD) Yuji Tanaka, Ayumi Kurihara, Naoki Edo, Hirotsugu Kawamoto, Koji Morita, Hisanori Dambara National Defense Medical College, Tokorozawa, Japan; National Defense Medical College, Tokorozawa, Japan [bold]Background:[/bold] MMI, which is classified as a first line treatment of GD in Japan, is not necessarily used as such in other countries, partly because it has some serious adverse effects (WBC, liver or skin). Although they are known to be dose- and time- dependent, it is not wise to simply reduce the dose or duration, because failure in initial treatment could increase the risk of thyroid storm or [ldquo]drop-out[rdquo] from treatment. Through efforts to overcome these problems, we have developed a method that can more safely induce initial relief of thyrotoxicosis.[br][bold]Patients: [/bold]Diagnosis was made upon FT4, TSH and TBII/TSAb. Both fresh (drug na[iuml]ve) and recurrent cases were employed. Pregnant or heart failure was excluded.[br][bold]Methods: [/bold]MMI was administered as follows: (1) initial dose was calculated by a simple formula: (FT4-1)^0.5x(body weight(kg))/5 (mg/day) and was started, (2) then the dose was cut in half (or 15 mg/day maximal) at day 7. Clinical symptoms (including side effects), FT4, TSH, CBC and blood chemistry were followed.[br][bold]Results: [/bold]41 patients were assigned (F33, M8 mean 43yo). Data at first visit were FT4 5.9+-2.6(ng/dl) and MMI 22+-5.5(mg/day). At one week, FT4 decreased to 60+-20%, and the reduced dose of MMI was 9.5+-3.0(mg/day). At four weeks, FT4 was 43+-18%. About symptoms, 63% of patients improved in one week and 83% in four weeks. Four patients needed to increase MMI (by approx. 50%). In terms of side effects, 6 had slight itchiness, 3 had rash, 1 had general fatigue and 2 had edema or muscle spasm. Serious side effects such as agranulocytosis, liver dysfunction were not observed, and all could continue MMI.[br][bold]Discussion:[/bold] At the initial phase of GD treatment, our method improves FT4 and symptoms successfully without serious side effects. We speculate the reason was: 1. the amount calculated from FT4 were sufficient for MMI to exert its initial action after delivery into the gland; 2. the concentration of MMI in plasma that determines its side effects were low enough during the [ldquo]risky[rdquo] period because the dose was cut in half at day 7. One might criticize that our method needs more time to normalize FT4 than conventional ways, but this slowness on the other hand decreases the risk of edema or muscle spasm, which sometimes confuse treatment course.[br][bold]Conclusion:[/bold] By using our [ldquo]On [amp] Half[rdquo] method, MMI should become much safer and more effective tool to initiate treatment of GD.[br][br]Nothing to Disclose: YT, AK, NE, HK, KM, HD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 293 309 2300 MON-410 PO40-01 Monday 1969 2012


1965 ENDO12L_MON-411 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Autoimmune Thyroid Disease, Insulin Resistance and Cardiovascular Risk Factors Celestino Neves, Camila Dias, Miguel Pereira, Carmo Palmares, Oksana Sokhatska, Marta Alves, Davide Carvalho, Luis Delgado, Jose L Medina S[atilde]o Jo[atilde]o Hospital, Faculty of Medicine, University of Porto, Porto, Portugal; S[atilde]o Jo[atilde]o Hospital, Faculty of Medicine, University of Porto, Porto, Portugal; S[atilde]o Jo[atilde]o Hospital, Faculty of Medicine, University of Porto, Porto, Portugal Objective: To examine whether Graves[apos] disease (GD) and autoimmune thyroiditis (AIT) are associated with insulin resistance and other cardiovascular (CV) risk factors. Subjects and Methods: We recorded thyroid function tests, BMI, insulin resistance markers comprising the Homeostasis Model Assessment for insulin resistance (HOMA-IR and HOMA-B), the Quantitative Insulin Sensitivity Check Index (QUICKI), HISI (Hepatic Insulin Sensitivity Index), WBISI (Whole-Body Insulin Sensitivity Index), IGI (Insulinogenic Index) and the levels of total cholesterol (TC), HDL, LDL-cholesterol, triglycerides (TG), apolipoprotein B (ApoB), ApoA1, lipoprotein (a) (Lp[a]), homocysteine, CRP (C-reactive protein), folic acid and vitamin B12 levels, in a total of 213 patients, 93.4% female, with a BMI of 28.8[plusmn]4.9 Kg/m2 and a mean age of 46.8[plusmn]15.2 years. There were 48 patients with GD and 180 patients with AIT. The patients with GD and with AIT were treated, in order to normalize T3, T4 and TSH levels. A 75-g OGTT was performed in the morning (before 11 AM), and blood samples were obtained every 30 min for 120 min for measurements of plasma glucose, insulin, and C-peptide. Statistical analysis was performed with ANOVA and Pearson[apos]s correlations test. Results are expressed as mean [plusmn] SD. A two-tailed p value [lt] 0.05 was considered significant. Results: There were no significant differences between PCR, Lp(a), homocysteine and insulin-resistance levels in patients with AIT and GD. We found that patients with AIT had significantly higher levels in LDL (128.1[plusmn]30.9 vs 117.6[plusmn]32.2 mg/dl, p=0.42) and ApoB (99.7[plusmn]24.1 vs 88.9[plusmn]29.2 ml/dl, p=0.007) Within the AIT group we found significant correlations between anti-TPO and TC (R=0.17; p=0.02), HDL (R=-0.36; p=0.01), LDL (R=0.17; p=0.03), and ApoA1 (R=-0.45; p[lt]0.01). We also found correlations between HDL and HOMA-B (R=0.15; p[lt]0.05) and HOMA-IR (R=-0.20; p[lt]0.01), and between Lp(a) and anti-TG (R=-0.17; p=0.03). In the GD group we found significant correlations between PCR and ApoB (R=0.36; p=0.02), HOMA-IR (R=0.39; p[lt]0.001), and WBISI (R=-0.35; p=0.02). In this group, HOMA-IR was correlated with TG (R=0.35; p=0.02), and ISI with anti-TG (R=0.52; p[lt]0.001). Conclusion: In our study, patients with AIT had significantly higher levels of LDL and ApoB than patients with GD. Treated autoimmune thyroid disease is intrinsically linked to variables of insulin resistance and other CV risk factors.[br][br]Nothing to Disclose: CN, CD, MP, CP, OS, MA, DC, LD, JLM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1323 309 2301 MON-411 PO40-01 Monday 1970 2012


1966 ENDO12L_MON-412 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Hyperprolactinemia: A Risk Factor of Thyroid Disease in Patients with Prolactinoma Muyesser Sayki Arslan, Oya Topaloglu, Esra Tutal, Basak Karbek, Bekir Ucan, Ilknur Ozturk Unsal, Askin Gungunes, Evrim Cakir Ozkaya, Nujen Colak Bozkurt, Mustafa Ozbek, Erman Cakal, Mustafa Sahin, Tuncay Delibasi Diskapi Yildirim Beyazit Training and Research Hospital, Ankara, Turkey; School of Medicine, Ankara University, Ankara, Turkey [bold]Background: [/bold]Hyperprolactinemia has been reported to be associated with increased prevalence of ultrasonographic thyroid abnormalities and thyroid autoantibodies. The causal relationship between hyperprolactinemia and thyroid diseases has not yet well defined. The aim of this investigation was to evaluate the effects of hyperprolactinemia on thyroid function, volume and nodularity in a mild iodine-deficient area.[br][bold]Patients and Methods: [/bold]Fifty-eight (58) hyperprolactinemic subjects, 18 men and 40 women, aged 38.9[plusmn]10.6 years with newly diagnosed prolactinoma (mean prolactin 98.7[plusmn]11.4 mg/l) were recruited from our outpatient clinic. Fifty-seven healthy subjects, without any known disease, of similar age, gender and body mass index were included as a control group. Serum prolactin, thyroid stimulating hormone (TSH), free tri-iodothyronine (free T3) and thyroxine (free T4), thyroidal microsome (anti-TPO) and antithyroglobulin antibodies levels were evaluated and the thyroid volume was calculated with the using of ultrasound examination (Hitachi EUB-7000 HV).[br][bold]Results: [/bold]TSH levels were significantly lower in patients with prolactinoma than in controls (1.84 [plusmn]0.99 uIU/ml vs 2.26[plusmn]1.24 uIU/ml, respectively, p=0.007). Frequencies of positive anti-TPO were significantly (p=0.001) higher in prolactinoma subjects whereas there were no significant difference in antithyroglobulin antibodies (p=0.25). Also the percentage of patients thyroid nodules were significantly higher in patients with prolactinoma than in controls (9.2% vs 32.6%, respectively, p=0.05). There was no thyroid cancer according to FNA results. Positive correlation was found between thyroid volume and prolactin level (r =0.544, p=0.001). Prolactin has significant effect on total volume according to step-wise multiple linear regression analysis (adjusted R square is 0.268).[br][bold]Conclusion: [/bold]The results suggest that patients with prolactinoma have significantly increased thyroid volume, thyroid autoimmunity and nodule prevalance. Our data indicate that hyperprolactinemia is a risk factor for thyroid autoimmunity and nodular thyroid disease.[br][br]Nothing to Disclose: MSA, OT, ET, BK, BU, IOU, AG, ECO, NCB, MO, EC, MS, TD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 758 309 2302 MON-412 PO40-01 Monday 1971 2012


1967 ENDO12L_MON-413 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Presence of Adenosine A2a Receptor in Thyrocytes and Its Involvement in the VEGF Expression Induced by Graves IgG Lin Zhang, Nannan Sun, Meng Ren, Ling Gao, Jiajun Zhao Provincial Hospital Affiliated to Shandong University, Jinan, China; Shandong Academy of Clinical Medicine, Jinan, China; Provincial Hospital Affiliated to Shandong University, Jinan, China Background:[br]Graves[apos] disease (GD) is an autoimmune disease. Thyroid angiogenesis is proved to be correlated with thyroid function and goiter volume in GD. A central role of A2a receptor (A2aR) has been reported in many extrathyroidal tumors. However, it was not clear that whether A2aR exists in thyroid and whether, if expression, there is an association with autoantibodies of GD in angiogenesis. Our aim was to detect the presentation of A2aR in thyroid and its possible role in the induction of VEGF, the most potent angiogenic factor, under stimulation of GD IgG.[br]Methods:[br]A2aR presentation on both FRTL-5 cells and primary human normal thyrocytes was confirmed by PCR, western-blotting, immunohistochemistry and immunofluorescence, respectively. Cells were treated with CGS21680 (A2aR specific agonist) or crude IgG from the serum of healthy individual and untreated GD patients in the present or absent of ZM241385 (A2aR specific antagonist) for 48h. VEGF expression was tested by both RT-PCR and ELISA. The upstream regulators of VEGF such as PGC-1[alpha], HIF-1 [alpha] and p-Akt were evaluated by western blotting, respectively. Serum TRAb levels and goiter volumes of the patients were measured respectively.[br]Results:[br](1) Thyrocytes [italic]per se[/italic] had mRNA and protein expression of A2aR. The proteins located on the cell membrane. (2) The VEGF splicing isoforms, 188, 164, 144, 120, and 189, 165, 145, 121 were detected, respectively. VEGF188 and 164 were enhanced by approximate 2-fold (both[italic] p [/italic][lt] 0.05) when stimulation with 1 [mu]M CGS21680, accompany with a 23.3% increase in secreted VEGF contents and a 0.89% increase in intracellular VEGF contents (both [italic]p[/italic][lt]0.05). (3) Compared with the normal control and IgG from GD patients with smaller goiter, IgG from the patients with goiter volume larger than 40 cm[sup]3[/sup] increased the expression of VEGF188, 164, 144 and 120 by approximate 2-fold ([italic]p[/italic][lt] 0.05 for all) while ZM241385 crippled the effects. (4) Three VEGF transcription regulators were tested. CGS21680 increased nuclear PGC-1 [alpha] to about 2 times and GD IgG to about 1.5 times ([italic]P [/italic][lt] 0.05 for both). No influence was observed on HIF-1[alpha] and p-Akt. (5) The serum TRAb levels of GD patients were positively correlated with their thyroid volumes (R = 0.69, P[lt]0.01).[br]Conclusion:[br]Functional A2aR was expressed in thyrocytes and had the potential to modulate the expression of VEGF. GD IgG induced VEGF expression can be partly attribute to A2aR activation.[br][br]Sources of Research Support: National Natural Science Foundation of China(30672748).[br][br]Nothing to Disclose: LZ, NS, MR, LG, JZ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1190 309 2303 MON-413 PO40-01 Monday 1972 2012


1968 ENDO12L_MON-414 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Quercetin Inhibits CXC Chemokine Ligand 10 Secretion in Graves Ophthalmopathy Sena Hwang, Hyun Jung Lee, Dong Yeob Shin, Eun Jig Lee Brain Korea 21 Project for Medical Science, Institute of Endocrine Research, and Severance Integrative Research Institute for Cerebral [amp] Cardiovascular Disease, Seoul, Korea CXC chemokine ligand 10 (CXCL10) plays a pivotal role in the initial phases of autoimmune thyroid disease. A natural product quercetin has been shown to inhibit the inflammation process in Graves[apos] opthalomopathy (GO). However, there were no data about the effect of quercetin on secretion of CXCL10 in GO.[br]Experiments were carried out in primary cultured orbital fibroblasts from GO. ELISA was used to measure CXCL10 levels in culture supernatant. We have tested the role of quercetin on CXCL10 secretion in GO after simulation of this chemokine secretion with IFNg and TNF[alpha]. A peroxisome proliferator activated receptor (PPAR) [alpha] agonist, fenofibrate, a known inhibitor of CXCL10 secretion was also studied for comparison.[br]A dose-dependent inhibition by quercetin was observed on the cytokines-stimulated secretion of CXCL10 in GO cells. The potency of the quercetin used was maximum on the secretion of CXCL10 reaching about 67% of inhibition, comparison to 83% by fenofibrate. Quercetin was stronger inhibitor of CXCL10 secretion in GO than fenofibrate.[br]Our study shows that quercetin is potent inhibitor of the secretion of CXCL10, suggesting that quercetin may be involved in the modulation of the immune response in Graves[apos] disease.[br][br]Nothing to Disclose: SH, HJL, DYS, EJL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1529 309 2304 MON-414 PO40-01 Monday 1973 2012


1969 ENDO12L_MON-415 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) FOXP3 Polymorphisms Are Associated with Autoimmune Thyroid Disease Risk in Turner Syndrome Patients Bianca Bianco, Ieda TN Verreschi, Kelly C Oliveira, Caio Parente Barbosa, Monica VN Lipay Federal University of S[atilde]o Paulo, Sao Paulo, Brazil; Faculdade de Medicina do ABC, Santo Andre, Brazil [bold]BACKGROUND/AIM: [/bold]Turner Syndrome (TS) have increased risk for autoimmune diseases and the most common appear to be thyroid abnormalities. Recently, studies have associated [italic]FOXP3[/italic] gene, which regulates the activation of T cell and its functions, with homeostasis of the immune system and the development of autoimmune diseases. Here, we hypothesized a possible relationship between thyroid abnormalities and the[italic] FOXP3[/italic] polymorphisms in TS patients.[br][bold]METHODS OF STUDY:[/bold] Case-control study consisting of 84 Brazilian TS women (aged 4.8-55.5y) and a control group of 171 healthy fertile women (mean age 35.7[plusmn]4.9y) without history of autoimmune diseases. In TS group, 21.4% (n=18) had Hashimoto[apos]s thyroiditis. [italic]FOXP3[/italic] polymorphisms (rs3761548, rs3761549, rs2232368, rs2232366 and rs2280883) were identified by [italic]TaqMan[/italic] PCR. Chi-square was used to compare allele and genotype frequencies between groups and the association between the combined genotypes was assessed by haplotype analysis. Statistical significant values were corrected by Bonferroni method.[br][bold]RESULTS: [/bold]Single-marker analysis revealed a significant association between rs3761549 polymorphism and TS (p=0.005, OR=2.42, CI 95%=1.33-4.42). When we compared only TS with Hashimoto[apos]s thyroiditis and controls there was a statistically significant difference (p=0.003, OR=3.96, CI 95%=1.62-9.67). Considering the rs2280883 polymorphism, no difference was found between the TS and controls. Comparing only TS with Hashimoto[apos]s thyroiditis and controls, a statistically significant difference was observed (p=0.029, OR=0.22, 95% CI = 0.06 to 0.74), however, the odds ratio does not show such significance. Considering rs3761548 polymorphism, no difference was found between TS and controls, even considering only patients with Hashimoto[apos]s thyroiditis. Similar results were found to rs2232368 and rs2280883 polymorphisms. Combined genotype analysis identified a haplotype [quot]ATGAG[quot] associated with autoimmunity in TS (4.3% in cases and 1.0% in controls), p=0.016. In contrast, haplotype [quot]CCTGA[quot] was associated with lower risk of autoimmunity (53.5% in cases and 64.6% in controls), p=0.015. After Bonferroni correction, only rs3761549 polymorphism remains statistically significant.[br][bold]CONCLUSION:[/bold] To our knowledge, this is the first study that associated [italic]FOXP3[/italic] polymorphisms and thyroid abnormalities in TS. The results showed association between rs3761549 polymorphism and Hashimoto[apos]s thyroiditis in Brazilian TS patients tested.[br][br]Sources of Research Support: CAPES for granting student Bianca Bianco a postdoctoral scholarship.This work was supported by the grants No. 2009/05250-0 from FAPESP.[br][br]Nothing to Disclose: BB, ITNV, KCO, CPB, MVNL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 49 309 2305 MON-415 PO40-01 Monday 1974 2012


1970 ENDO12L_MON-416 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) The Prevalence of Thyroid Disorders and the Positive Rates of Thyroid Autoantibodies in North Korean Refugees in South Korea Compared to South Koreans Joo Hyung Kim, Sin Gon Kim, Dong Seop Choi, Sei Hyun Baik, Kung Mook Choi, Nan Hee Kim, Ji A Seo, Hee Young Kim, Hye Jin Yoo, Sae Jung Yang, Yoon Jung Kim, Hae Yoon Choi, Chae Ryung Eun, Nam Hoon Kim, Jae Hee Ahn Korea University Hospital, Seoul, Republic of Korea Thyroid dysfunction is one of the most common endocrine disorders and its prevalence and pattern depend on ethnic and environmental factors including iodine intake. North Korean refugees are the best subjects to identify the influence of environmental factors in disease outcome because they have the same genetic background with South Koreans, but at the same time they have been exposed to different environments for a long time. This study is aimed to examine and compare the prevalence patterns of thyroid disorders between North Korean refugees(NKR) in South Korea and South Koreans(SK).[br]The intended sample was a total of 442 North Korean refugees aged 30 or more residing in Seoul. A total of 661 SK who were age and gender matched to NKR counterpart were also selected from a group who visited Korean University Medical Center for health check-up. Serum thyrotropin(TSH), free thyroxine(FT4) and thyroid autoantibodies (anti-thyroperoxidase antibody[TPOAb] and anti-thyroglobulin antibody[TgAb]) were measured from each population.[br]The mean serum TSH levels were not significant differences between two groups. But serum TSH levels were higher in SK males than NK males (median value 1.64 vs. 1.39). The prevalence of subclinical hyperthyroidism was similar between two groups (NKR vs. SK; 1.6% vs. 1.5%, p=0.918) and was not significantly different by age or sex. And the prevalence of subclinical hypothyroidism also was similar (NKR vs. SK; 7.5% vs. 8.9%, p=0.402). But in male, the prevalence of subclinical hypothyroidism was higher in SK than NKR, probably due to a smaller sample size in NKR male (NKR vs SK; 2.1%[2/95] vs. 10.2%[17/166], p=0.015). Overall, the positive rate of autoantibodies (either TPOAb or TgAb) was significantly higher in SK (23%) than NKR(9.4%)(p=[lt]0.001). Especially, the positive rate of TgAb in SK females was high. There was no overt hyperthyroidism or hypothyroidism in both groups.[br]The positive rates of autoantibodies were higher in SK than NKR. However, overall prevalence of subclinical thyroid dysfunction was similar between two groups. Further study is needed to find out why the prevalence of thyroid dysfunction was not different despite of different positive rate of autoantibodies.[br][br]Nothing to Disclose: JHK, SGK, DC, SHB, KMC, NHK, JAS, HYK, HJY, SJY, YJK, HYC, CRE, NHK, JHA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1501 309 2306 MON-416 PO40-01 Monday 1975 2012


1971 ENDO12L_MON-417 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Association of MICA Alleles with Autoimmune Thyroid Disease in Korean Children Won Kyoung Cho, Ji Hoon Kim, Kyoung Soon Cho, So Hyun Park, Seung Hoon Hahn, Min Ho Jung, Hee-Baeg Choi, Tai-Gyu Kim, Byung Kyu Suh College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea; College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea; College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea Backgrounds: Major histocompatibility complex (MHC) class I chain-related gene A (MICA) located only 46kb centromeric of the HLA-B gene, has been identified within the class I region on chromosome 6. Many reports relating the association of MICA polymorphism with diseases have been published. However, data regarding the association with autoimmune thyroid disease (AITD) were lacking. In the present study, we aimed to assess the MICA polymorphism and the association with AITD in Korean children. Methods: Seventy-two patients with AITD [Hashimoto disease (HD): 31, Graves disease (GD): 41] were recruited. We analyzed MICA polymorphism by PCR-SSP, and compared with those of 69 normal control. Results: In AITD, the allele frequencies of MICA*010([italic]P[/italic]=0.003) was higher and those of MICA*004([italic]P[/italic]=0.018) and MICA*008([italic]P[/italic]=0.004) were lower than control. In GD, those of MICA*010([italic]P[/italic]=0.004) was higher than control and those of MICA*008([italic]P[/italic]=0.003) was lower than control. Between HD and control group, there were no significant differences in allele frequencies. In AITD, MICA*010-HLA-A*02, MICA*010-HLA-B*46, MICA*010-HLA-Cw*01 and MICA*010-HLA-DR*08 were the most common two-locus haplotypes of MICA and HLA. When compared the relative strength of disease associations of MICA*010 and HLA-B*46 with diseases, those of two alleles were differ. The patients who did not have thyroid-assocated opthalmopathy (TAO) showed higher frequencies of MICA*010 and lower frequencies of MICA*008 compared to normal control group. Conclusion: It is possible that MICA*010 is positively associated with AITD, independent of HLA-B*46 and negatively associated with TAO in Korean children.[br][br]Nothing to Disclose: WKC, JHK, KSC, SHP, SHH, MHJ, H-BC, T-GK, BKS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 521 309 2307 MON-417 PO40-01 Monday 1976 2012


1972 ENDO12L_MON-418 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) [italic]TYK2[/italic] Polymorphisms in Turner Syndrome Bianca Bianco, Ieda TN Verreschi, Kelly Cristina Oliveira, Carla Peluso, Caio Parente Barbosa, Monica VN Lipay Universidade Federal de S[atilde]o Paulo, S[atilde]o Paulo, Brazil; Faculdade de Medicina do ABC, Santo Andre, Brazil [bold]Background: [/bold]Turner Syndrome (TS) have increased risk for autoimmune diseases and the most common appear to be thyroid abnormalities. Tyrosine kinase 2 ([italic]TYK2[/italic]) is a gene that have crucial importance in the etiology of autoimmune and inflammatory diseases. Many polymorphisms of the [italic]TYK2[/italic] gene have been identified, and recently, a number of case[ndash]control studies were conducted to investigate the association of rs34536443 and rs2304256 polymorphisms with autoimmune and inflammatory diseases, with conflicting results. Here, we hypothesized a possible relationship between Hashimoto[acute]s disease and [italic]TYK2[/italic] polymorphisms in TS patients.[br][bold]Methods:[/bold] We performed a case-control study comprising 100 TS women (aged 4.8-55.5y) and 217 fertile women (mean age 39.7[plusmn]3.2y) without history of autoimmune diseases, as controls. In TS group, 21.4% (n=18) had Hashimoto[acute]s disease. [italic]TYK2[/italic] polymorphisms (rs34536443/C-G/Pro-Ala at exon 23 and rs2304256/A-C/Val-Phe at intron 18) were identified by [italic]TaqMan[/italic] real time PCR. The results were analyzed statistically and a p-value [lt]0.05 was considered significant.[br][bold]Results: [/bold]Single-marker analysis revealed that [italic]TYK2[/italic] rs34536443 was significantly associated with protection against thyroid autoimmunity, even considering only TS patients with Hashimoto[acute]s disease, since the polymorphic allele was more frequent in controls than in cases (p=0.012, polymorphic allele is present in 2.5% of Turner Syndrome and 8% in the control group). Considering rs2304256 polymorphism no association was found (p=0.423), even considering only patients with HD. All groups were in Hardy-Weinberg equilibrium to both polymorphisms studied. Haplotype analysis of two [italic]TYK2 [/italic]polymorphisms identified a haplotype [apos][apos]CC[apos][apos] associated protection against thyroid autoimmunity (p=0.028, frequency of 8% in Turner Syndrome patients and 40% in control group).[br][bold]Conclusion: [/bold]This is the first study to report a possible association between [italic]TYK2[/italic] polymorphisms and autoimmune thyroid disease in Turner Syndrome. These findings require replication in other populations but suggest the [italic]TYK2[/italic] rs34536443 polymorphism is associated with protection against Hashimoto[acute]s disease in Brazilian Turner syndrome patients studied.[br][br]CAPES for granting student Bianca Bianco a postdoctoral scholarship. This work was supported by the grants No. 2009/05250-0 from FAPESP.[br][br]Sources of Research Support: Oliveira RM, et al. Y chromosome in Turner syndrome: review of the literature. Sao Paulo Med J 2009;127, 373. Bianco et al. The possible role of genetic variants in autoimmune-related genes in the endometriosis development. Hum Immunol 2011 [Epub ahead of print]. J[oslash]rgensen KT, et al. Autoimmune diseases in women with Turner[apos]s syndrome. Arthritis Rheum 2010;62:658-66. Tao JH, Zou YF, Feng XL, Li J, Wang F, Pan FM, Ye DQ. Meta-analysis of TYK2 gene polymorphisms association with susceptibility to autoimmune and inflammatory diseases. Mol Biol Rep 2011;38(7):4663-72.[br][br]Nothing to Disclose: BB, ITNV, KCO, CP, CPB, MVNL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 938 309 2308 MON-418 PO40-01 Monday 1977 2012


1973 ENDO12L_MON-419 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Detection of Thyroid-Blocking and Stimulating Immunoglobulins in Patients with Autoimmune Thyroid Disease Paul D Olivo, Jaekyung Kim, Tanja Diana, Rebecca Klasen, Michael Kanitz, Yunsheng Li, George J Kahaly Diagnostic Hybrids, Inc, Athens, OH; Gutenberg University Medical Center, Mainz, Germany [bold]Background and aims: [/bold]Novel bioassays were developed for the detection of thyroid-stimulating (TSAb) and thyroid-blocking autoantibodies (TBAb). Both bioassays are based on a cell line (CHO-MC4) expressing a chimeric TSH-receptor (TSHR). These bioassays were used to measure the functional activity of anti-TSHR autoantibodies in patients with autoimmune thyroid disease (AITD).[br][bold]Methods[/bold]: TBAb was measured in a newly developed assay in which induction of luciferase by a set concentration of bovine TSH (bTSH) is compared to the luciferase induced by bTSH in the presence of patient sera. Results are expressed as percent inhibition. Positive percent inhibition indicates blocking activity; negative percent inhibition indicates stimulatory activity. Fifty patients with AITD were tested in both the TSAb and TBAb assays. TSAb was measured with an FDA-cleared test (Thyretain[reg], DHI, Athens, OH) according to the manufacturer[apos]s recommendation. Results were expressed as specimen-to-reference ratio as a percentage (SRR %).[br][bold]Results[/bold]: Three hundred and one normal control sera were tested in the TBAb bioassay to set the assay cut-off. Results ranged from -59% to +38% with a mean of -4% [plusmn] SD (standard deviation) of 21%. Using two SD from the mean the cut-off was 40%. A retrospective cross-sectional trial using 50 control sera and 50 sera from patients with AITD was performed. The control sera ranged from [ndash]16% to +37 with a mean of 6% [plusmn] 11%. Sera from patients with AITD had results that ranged from -157% to +108%. Sera that were positive TSAb bioassay had negative % inhibition of -28% to -157% in the TBAb bioassay. Fifteen of 50 (30%) patients (seven hypothyroid, two with serum baseline TSH above 100 mU/L, nine female, 13 with autoimmune thyroiditis and two with Graves[apos] disease, aged 3 months to 78 years) had positive inhibition [gt]40%, ten of which were [gt]80%. Of these 15 TBAb positive patients, 11 were positive for thyroid binding inhibiting immunoglobulins (automated electrochemiluminescence TBII assay, Elecsys TRAb, Roche). Review of the clinical history of these TBAb positive patients revealed the TBAb results were consistent with the clinical presentation.[br][bold]Conclusions:[/bold] Bioassays for thyroid-stimulating and blocking antibody are useful for evaluating patients with AITD. Identifying the functional type of anti-TSHR autoantibody that is present in these patients can help explain the clinical presentation and improve the management of these patients.[br][br]Disclosures: PDO: Employee, Diagnostic Hybrids, Inc. JK: Employee, Diagnostic Hybrids, Inc. RK: Employee, Diagnostic Hybrids, Inc. YL: Employee, Diagnostic Hybrids, Inc. GJK: Research Funding, Diagnostic Hybrids, Inc. Nothing to Disclose: TD, MK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1456 309 2309 MON-419 PO40-01 Monday 1978 2012


1974 ENDO12L_MON-420 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Correlation of 25 OH D3 Levels with Thyroid Autoantibodies in Greek Population Evangelia Zapanti, Maria Prokopiou, Georgia Kassi, Nektarios Benetatos, Helen Theohari, Helen Karga, Argyris Papadakis, Maria Christou, Athanasios Karahalios Laiko Hospital, Athens, Greece; Alexandra General Hospital, Athens, Greece The role of vitamin D as an immune modulator is now well recognized. Besides bone mineral omeostasis, vitamin D deficiency has been associated with several autoimmune diseases.The nature of the interaction between vitamin D and the immune system became evident after the discovery of vitamin D receptor (VDR) in immune cells such as activated CD4+,CD8+ T cells, B cells and APCs. In vitro studies have shown that vitamin D supports a THh1 shift to Th2 in CD4+ cells, however, in vivo studies have failed to prove a predominance of Th2 immunity. Apparently, the in vivo role of vitamin D on the immune system is more complex.[br]The aim of this study is to assess the levels of vitamin D in respect to thyroid autoimmunity status in Greek population[br]Method[br]A total of 1997 subjects consulting for various non-thyroid related reasons, aged 16-88 (median age 62 y) were recruited in an outpatient endocrine clinic over a period of 3 years. Fasting blood samples were drawn for the estimation of 25OHD3, anti-Tg and anti TPO antibodies levels. In another sample of 51 patients suffering Graves disease, aged 55[plusmn]12 y, TRAB and 25 OH D3 levels were estimated. Chemofluorescence was used to estimate antibody levels.[br]Statistics[br]Linear correlation and linear regression analyses. Non-detectable levels were replaced by values corresponding to 2 SD below detection levels[br]Results[br]Anti Tg and Anti TPO levels fluctuate between non-detactable to 30,000 IU/ml and 4,500 IU/ml respectively. A strong positive correlation was detected between anti TG and anti TPO values (r=0.458, P[lt]0.001, n=1997). No correlation was found between 25 OH D3 and anti TG or anti TPO. Strong positive correlation was detected between TRAB and 25 OH D3 levels in the Graves patient sample (r=0.356, P[lt]0.01, n=52).[br]Conclusion[br]The positive correlation between 25OHD3 levels and TRAB suggests a role of Vitamin D in GRAVES disease (Th2 immunity related).[br][br]Nothing to Disclose: EZ, MP, GK, NB, HT, HK, AP, MC, AK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1270 309 2310 MON-420 PO40-01 Monday 1979 2012


1975 ENDO12L_MON-421 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Incidental Diffuse Thyroid [sup]18[/sup]F-FDG Uptake Is Favorable Prognostic Factor in Advanced Breast Cancer Yang Seon Lee, Won Jin Kim, Min Jung Bae, Ji Hyun Kang, Bo Gwang Choi, Yun Kyung Jeon, Snag Soo Kim, Bo Hyun Kim, Yong Ki Kim, In Ju Kim Pusan National Univeristy Hospital, Busan, Korea; Kim Young Ki Internal Medicine Clinic, Busan, Korea [bold]Background and Aim: [/bold]Patients with breast cancer have a relatively high prevalence of diffuse thyroid fluorodeoxyglucose (FDG) uptake related to thyroid autoimmunity. It is postulated that the presence of thyroid autoimmunity has prognostic implications for breast cancer. The aim of this study was to evaluate the prognostic values of incidental diffuse thyroid F-18 FDG uptake in breast cancer.[br][bold]Patients and Methods:[/bold] We evaluated 564 patients that underwent breast surgery for primary breast cancer between January 2006 and December 2009. All patients had available FDG PET-CT imaging more than two times. Patients were divided into 2 groups according to the diffuse thyroid FDG uptake.[br]Main outcome measure was disease-free survival of breast cancer.[br][bold]Results: [/bold]Of the 564 patients, 108 (19.1%) patients showed diffuse thyroid uptake on preoperative and/or postoperative PET-CT. The median follow-up period was 36.0 months (range, 1.0[ndash]77.0 months). There was higher levels of serum TSH in patients with thyroid FDG uptake compared with in those without uptake ([italic]P [/italic]= 0.001). Both TPO and Tg antibody titers were higher in patients with thyroid FDG uptake than those without uptake ([italic]P [/italic][lt] 0.001in both). Of the 108 patients with thyroid FDG uptake, 5 patients had a recurrence of breast cancer, whereas 85 patients without uptake had a recurrence (log rank statics = 12.28, [italic]P [/italic][lt] 0.001). The association of diffuse thyroid FDG uptake with tumor recurrence was not significant in multivariate analysis in patients with early stage (HR = 0.26; 95% CI: 0.06[ndash]1.10; [italic]P[/italic] = 0.067). However, the association of diffuse thyroid FDG uptake with breast cancer recurrence was remained statistical significance in multivariate analysis adjusting for several prognostic variables (HR = 0.19; 95% CI: 0.57[ndash]0.62; [italic]P[/italic] = 0.006).[bold]Conclusions:[/bold] Incidental diffuse thyroid FDG uptake related to autoimmune thyroiditis was independently favorable prognostic factor for advanced breast cancer. These findings support evidence that thyroid autoimmunity might be associated with a beneficial effect on outcome of breast cancer.[br][br]Nothing to Disclose: YSL, WJK, MJB, JHK, BGC, YKJ, SSK, BHK, YKK, IJK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 349 309 2311 MON-421 PO40-01 Monday 1980 2012


1976 ENDO12L_MON-422 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Primary Radioactive Iodine Therapy Is Superior to Delayed Therapy for Thyrotoxicosis Saad Sakkal Metabolic Care Center, Aleppo, Syrian Arab Republic Objective: Radioactive iodine has been commonly used in controlling thyrotoxicosis for years. But, when is the best time to use it? At diagnosis or after quieting down the acute florid symptoms with thiamazole/PTU?[br]Methods: We prospectively compared two different timing of using iodine to effect clinical manifestations: either at the time of diagnosis or after using Methimazole/PTU first to quiet down the clinical presentation followed by radioactive iodine within 3 months. Thyrotoxicosis symptoms were controlled by Atenolol in both groups from the first day of diagnosis and both were strictly advised to raise the head of bed to prevent ophtalmopathy. We report the results of the first 200 patients who were assigned alternatively to either method. Clinical Effects measured included Pulse, Bp, weight, thyroid tests and subjective feeling of palpitation, tiered, hot, headache, and sleep. In addition, we determined final recovery from thyrotoxicosis or complications of therapy (including CHF, AF, ophtalmopathy, or neurologic complications).[br]Results: There was no significant difference in Symptoms number improved, sleep or weight at one month, but T4 normalization was better in anti-thyroid drugs therapy (%45 Vs65) and not different at 6 months(%70 Vs. 69). However patients reported much more satisfaction with primary radioactive iodine therapy than delayed therapy (%80 VS%60) because of its faster recovery/return to normal life, ease/convenience and much less expense involved at 6 months (16000 Vs29000). In general there was also less clinical symptoms (feeling of palpitation, fatigue, hot or headache/cerebral symptoms) at 6 months (%11 Vs32).Better recovery at 24 months (%83 Vs71) and less repeat of I* or persistent need for PTU/Methimazole (%10 Vs23). Also complications were much less with primary therapy (%12 Vs19). All differences were significant at P value of 0.001 or more. Pulse and Bp control was well controlled with Atenolol.[br]Conclusion: Primary radioiodine therapy is much better choice in controlling thyrotoxicosis than delayed therapy after PTU/Methimazole use. It has better control of Pulse, Bp, palpitation, fatigue, warm feeling and cerebral symptoms at 6 months. It has better recovery rate, less repeated radioiodine dose, less persistent need for PTU/Thiamazole at 2 years and much less expense and complications.[br][br]Nothing to Disclose: SS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 148 309 2312 MON-422 PO40-01 Monday 1981 2012


1977 ENDO12L_MON-423 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Determining Autoantibodies to Thyroid Peroxidase and Thyroglobulin in a Pediatric Population Liliana Karina Silvano, Alejandra Paez, Gabriela Maria Sobrero, Mariana Ochetti, Gisel Schvab, Cintia Tariffa, Graciela Testa, Adriana Boyanovsky, Silvia Edith Martin, Mirta Beatriz Miras, Liliana Noemi Munoz Hospital de Ni[ntilde]os de la Santisima Trinidad, C[oacute]rdoba, Argentina [bold]Introduction and Aim.[/bold][br]Autoantibodies to thyroid Peroxidase and Thyroglobulin are markers of thyroid autoimmunity and can be quantified by different assays. It is necessary to establish reference values for each method in paediatric populations for an effective use in the study of autoimmune diseases (AD). Aims: To determine the cut-off levels of TPOAb and TgAb for two methods in healthy children. To establish the diagnostic concordance and the prevalence in different AD.[br][bold]Materials and Method.[/bold][br]We determined TPOAb and TgAb in 616 samples using ECLIA-Roche and CLIA-Siemens. We analyzed 229 healthy children and 387 patients with associated autoimmune diseases. Cut-off levels, diagnostic concordance and prevalence were determined.[br][bold]Results.[/bold][br]The levels of TPOAb and TgAb (IU/ml) in percentile 95, 97 and 97.5 were: TPOAb CLIA 11.8, 17.6 and 18.2; TPOAb ECLIA 26.1, 31.7 and 43.1; TgAb CLIA 10.0, 21.7 and 22.6; TgAb ECLIA 35.0, 51.3 and 85.7 respectively. TPOAb and TgAb were more prevalent in Hyperthyroidism, Turner Syndrome and Diabetes Mellitus. We observed significative differences in the diagnostic concordance of TgAb measurement (p=0.0003). The highest TSH levels were observed with positive autoantibodies for each method. (p[lt]0.01).[br][bold]Conclusions.[/bold][br]Our results allowed us to establish cut-off levels for the methods used in paediatric patients and confirmed the high prevalence of these antibodies in autoimmune diseases.[br][br]Nothing to Disclose: LKS, AP, GMS, MO, GS, CT, GT, AB, SEM, MBM, LNM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1936 309 2313 MON-423 PO40-01 Monday 1982 2012


1978 ENDO12L_MON-424 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Serum Thyroid-Stimulating Hormone Levels Are Associated with Lipid Parameters Independent of Thyroid Hormones in Euthyroid Subjects Furong Wang, Yinyin Tan, Chenggang Wang, Xu Zhang, Jiajun Zhao Provincial Hospital Affiliated to Shandong University, Jinan, China; Shandong University of Traditional Chinese Medicine, Jinan, China; Shandong University of Traditional Chinese Medicine, Jinan, China [bold]Context and Objective:[/bold] Dyslipidemia in thyroid dysfunction has always been attributed to changes of thyroid hormone levels. We hypothesized that thyroid-stimulating hormone (TSH) may play an important role in lipid metabolism independent of thyroid hormones. This question was addressed by studying the relationship of serum TSH levels with lipid profiles after controlling for FT3, FT4, TT3, TT4 and non-thyroid factors relevant to lipid metabolism in euthyroid Chinese subjects.[br][bold]Design and setting: [/bold]We conducted a cross-sectional study in subjects attending a routine health check-up at Provincial Hospital affiliated to Shandong University, China.[br][bold]Main outcome measures: [/bold]Serum lipid levels and thyroid functions were measured. General linear analysis was performed to determine if the impact of TSH on serum lipid levels is independent of thyroid hormone levels. Moreover, path analysis, an evolutionary multivariable regression technique commonly used in clinical epidemiology, was conducted to test whether there is a direct and/or indirect effect between serum TSH and TC levels. The odds ratios (Ors, 95% CI) for hypercholesterolemia in relation to the TSH categories were calculated using an adjusted logistic regression model.[br][bold]Results: [/bold]A total of 3,664 euthyroid subjects were finally analyzed. There was a significant linear trend towards higher TC ([italic]P[/italic] = 0.021) and TG ([italic]P[/italic] = 0.001) levels with increasing serum TSH levels within the reference range (0.27-5.5 mIU/L). These associations remained significant after adjusting for thyroid hormone levels, age, smoking, et al. Subgroup analysis showed the association between TSH and TC was stronger among older persons than younger individuals. Most importantly, the total effect of TSH on TC levels (total effect [sub]TC, TSH[/sub] = 0.05253; [italic]P[/italic] [lt] 0.05) includes a direct effect on TC levels (direct effect [sub]TC, TSH[/sub] = 0.05979; [italic]P[/italic] [lt] 0.05) and an indirect effect via thyroid hormones acting as an intermediate. Compared with subjects in the lower part of the reference range (TSH level = 0.27-0.61 mIU/L), the adjusted OR for hypercholesterolemia was 3.239 (95% CI = 1.392-7.538; [italic]P[/italic] [lt] 0.05) for those in the upper category (TSH level = 4.61-5.5 mIU/L).[br][bold]Conclusions:[/bold] The variation in normal TSH levels is related to the lipid components and hypercholesterolemia in euthyroid subjects, and it is independent of serum thyroid hormone levels. Our study suggests a potential new physiological role of TSH and the importance of controlling TSH in hypothyroid subjects.[br][br]Nothing to Disclose: FW, YT, CW, XZ, JZ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1325 309 2314 MON-424 PO40-01 Monday 1983 2012


1979 ENDO12L_MON-425 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Cardiac Autonomic Function in Patients with Subclinical Hypothyroidism Pramila Kalra, KM Kumar Prasanna, Vikram K Yeragani M S Ramaiah Medical College and Teaching Hospital, Bangalore, India; PRIMER, Bangalore, India; Wayne State University School of Medicine, Detroit, MI [bold]Background: [/bold]Patients with subclinical hypothyroidism may have higher incidence of coronary heart disease. [bold]Aim and objectives[/bold]: To compare linear and nonlinear measures of cardiac repolarization lability using beat-to-beat R-R intervals and QT intervals derived from the surface electrocardiogram during supine posture in patients with subclinical hypothyroidism and age and sex matched controls. [bold]Subjects and Methods[/bold]: We recruited 57 women (in the age range of 18-50 years) with subclinical hypothyroidism after an informed consent. Age and sex matched controls were also recruited. Patients who were pregnant or lactating, women on oral contraceptives or hormone replacement therapy, smokers and alcoholics, patients with body mass index (BMI) [gt] 30kg/m[sup]2[/sup], subjects on steroid therapy, high dose beta blockers, amiodarone, lithium were excluded from the study. Tests for cardiac autonomic function and vascular profiling were conducted in the morning time after the subjects had a light breakfast and they were advised not to drink any caffeinated beverages at least for 3 hours prior to the test. We compared linear measures of R-R interval variability using the surface electrocardiogram during supine posture. Spectral analysis was done to obtain VLF (very low frequency:0.003-0.04 Hz), LF(low frequency :0.04-0.15Hz) and HF(high frequency: 0.15-0.4Hz) components.The high frequency component represents vagal regulation (parasympathetic) and low frequency represents both parasympathetic and sympathetic regulation. The fractal dimension assessed the convolutedness and dimensionality of the heart rate time series. [bold]Results[/bold]: We recruited 57 women with a mean age of 31.7[plusmn] 8.8 years and 39 controls with mean age of 29.7[plusmn]7.1years(p=0.12).The high frequency component of RR(msec)/HR(bpm) was lower in cases as compared to controls(p=0.014). The ratio of LF/HF of R-R interval which is an index of sympathovagal tone was significantly more in cases compared to controls (p=0.037). The fractal dimension of RR interval was higher in cases compared to controls (p=0.021). [bold]Conclusions[/bold]: Patients with subclinical hypothyroidism have a decreased cardiac vagal function and possibly increased cardiac sympathetic activity.[br][br]Nothing to Disclose: PK, KMKP, VKY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 999 309 2315 MON-425 PO40-01 Monday 1984 2012


1980 ENDO12L_MON-426 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Peripheral Tissue Biomarkers in Resistance to Thyroid Hormone Carla Moran, Nadia Schoenmakers, Catherine Mitchell, Laura Watson, Greta Lyons, Phillippa Raymond-Barker, Kevin Taylor, David Halsall, VKK Chatterjee University of Cambridge, Cambridge, UK; Addenbrookes Hospital, Cambridge, UK Thyroid hormones (TH) regulate several peripheral tissue pathways including lipid metabolism and deiodinase enzyme activity and have also recently been reported to modulate trace element (copper, Cu) levels in rodents and humans (1). To assess their utility as tissue biomarkers of thyroid hormone action, we measured fasting circulating lipid and Cu levels, together with TH and its metabolite reverse T3 (rT3), in subjects with genetically-defined (defective TRbeta) Resistance to Thyroid Hormone, thyrotoxicosis and healthy controls.[br]Plasma copper levels were markedly raised in thyrotoxic patients (n=22, mean 21.8 [micro]mol/L) compared to RTH subjects (n=58, mean 17.07[micro]mol/L, p 0.05 x 10[sup]-3[/sup]) and healthy controls (n=107, 17.5[micro]mol/L, p=0.0012) and fell significantly following treatment (post-treatment mean 18.6 [micro]mol/L, p=0.048).[br]Baseline rT3 levels were significantly higher in the thyrotoxic patients (n=29, 94.6 ng/dl) than in RTH subjects (n=82, 68.1 ng/dl; p=0.012 x 10[sup]-2[/sup]) and healthy controls (n= 32, 24.4 ng/dl; p=0.009 x 10[sup]-8[/sup]) and declined markedly following treatment (preRx 62.4 ng/dl, postRx 25 ng/dl (p=0.03). RTH subjects exhibited lower FT3 levels than thyrotoxic subjects (13.3pmol/L vs 32.9 pmol/L; p=0.019 x 10[sup]-12[/sup]), such that their FT4/FT3 ratio is higher (2.8 vs 1.96; p=0.013 x 10[sup]-4[/sup]) but FT3/rT3 ratio is lower (0.21 vs 0.33; p=0.0151 x 10[sup]-5[/sup]) than in hyperthyroidism.[br]Comparison of lipid profiles showed significantly higher total cholesterol (5.18 mmol/L vs 4.85 mmol/L; p= 0.036), and LDL cholesterol (3.32 vs 2.93; p=0.003), but lower HDL (1.19 vs 1.49; p=0.06 x 10[sup]-6[/sup]) and higher triglycerides (1.50 vs 1.05; p=0.008 x 10[sup]-3[/sup]) in RTH subjects (n=93) versus healthy controls (n=113), resulting in higher cholesterol:HDL ratios in this disorder (4.63 vs 3.34; p= 0.018 x 10[sup]-7[/sup]).[br]Our observations, showing a lack of comparable elevation in circulating Cu, free T3 and rT3 in RTH versus thyrotoxicosis, substantiate the utility of these biomarkers in quantifying peripheral tissue refractoriness to TH action in RTH. Subjects with RTH exhibit mixed dyslipidaemia, with elevated total and LDLc but also raised triglycerides and low HDLc, similar to the adverse lipid profile seen in conventional hypothyroidism. Insulin resistance (2) and reduced vascular compliance (3) are recognised in RTH; we are determining whether their coexistence with dyslipidaemia and steatosis defines a subset of RTH patients at excess cardiovascular risk.[br][br](1) Mittag et al (2012) Serum copper as a novel biomarker for resistance to thyroid hormone. Biochemical Journal. Epub ahead of print; PMID 22220593. (2) Mitchell, C.S. et al (2010) Resistance to thyroid hormone is associated with raised energy expenditure, muscle mitochondrial uncoupling and hyperphagia. J Clin Invest, 120: 1345-54. (3) Owen, P. et al (2009) Augmentation index in resistance to thyroid hormone. Clin Endocrinol, 70: 650-54.[br][br]Sources of Research Support: Biomedical Research Centre; Wellcome Trust.[br][br]Nothing to Disclose: CM, NS, CM, LW, GL, PR-B, KT, DH, VKKC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2048 309 2316 MON-426 PO40-01 Monday 1985 2012


1981 ENDO12L_MON-427 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Thyroid-Stimulating Hormone, Independent of Thyroid Hormones, Is Positively Associated with the Serum Cholesterol Level in Patients with Coronary Heart Disease Chao Xu, Ling Gao, Jiajun Zhao Institute of Endocrinology, Shandong Academy of Clinical Medicine, Jinan, China; Institute of Endocrinology, Shandong Academy of Clinical Medicine, Jinan, China [bold]Background:[/bold] The relationship between thyroid stimulating hormone (TSH) and the lipid profile is contradictory because few studies have excluded the potential influence of the thyroid hormones (TH)[sup](1-6)[/sup]. Interestingly, [italic]in vivo[/italic] and [italic]in vitro[/italic] research by our laboratory on the function of TSH has shown that TSH, independent of TH, can upregulate the expression of HMGCR (the rate-limiting enzyme in cholesterol synthesis) and increase the cholesterol content in the liver[sup](7,8)[/sup]. Therefore, we hypothesized that TSH, independent of TH, would be positively associated with the serum cholesterol level.[br][bold]Methods:[/bold] A total of 1302 CHD patients diagnosed by coronary angiography were retrospectively studied. Thyroid function of the patients was measured twice, before hospitalization and the second day after hospitalization. The prevalence and distribution of thyroid dysfunction were analyzed first. To assess the impact of TSH on serum lipids, Pearson[apos]s correlation analysis was performed after adjustments for classic factors and TH. To calculate the extent of the effect of TSH on the serum cholesterol level, the partial least squares method and additional statistical methods were used.[br][bold]Results:[/bold] To ensure our study to have considerable strength, we used more stringent exclusion criteria. We ruled out the individuals with euthyroid sick syndrome and the patients enrolled our study were clinically stable. After the exclusions, a total of 568 patients (270 males and 298 females with a mean age of 63.56[plusmn]11.376 years) were selected. The prevalence of thyroid dysfunction among the patients was 18.66%, and the prevalence of hypothyroidism (15.32%) was higher than that of hyperthyroidism (3.34%). The prevalence of hypercholesterolemia was 13.5%, showing a linear and significant increase with elevations of the serum TSH levels (Pearson[apos]s Chi-squared test, linear trend 0.010, [italic]p[/italic] [lt] 0.05). Even after adjusting for confounding factors, such as sex, age, smoking status, fasting plasma glucose levels and TH, a significant positive impact of TSH on the serum total cholesterol (TC) level was revealed (r=0.095, [italic]p[/italic]=0.036). Each 1 mIU/L increase in the TSH level was linked to a 0.015580712 mmol/L elevation of the serum TC value.[br][bold]Conclusions:[/bold] In conclusion, we found a novel direct clinical effect of TSH that TSH, independent of TH, can increase the TC level in CHD patients. Although this is a very small effect, it is biologically and clinically significant.[br][br]1. Walsh JP et al., Arch Intern Med 2005; 165:2467-2472. 2. Roos A et al., J Clin Endocrinol Metab 2007; 92: 491-496. 3. A. IQBAL1 et al., J Intern Med 2006; 260: 53[ndash]61. 4. Neves C et al., Rev Port Cardiol 2008; 27; 1211-1236. 5. Cappola AR et al., JAMA 2006; 295: 1033-1041. 6. Vierhapper H et al., Thyroid 2000;10:981-984. 7. Tian L et al., Hepatology 2010; 52; 1401-1409. 8. Zhang W et al., J Cell Mol Med 2009;13; 4636-4642.[br][br]Sources of Research Support: The National Natural Science Foundation of China 81170794,30971409;the Natural Science Foundation ZR2009CZ009.[br][br]Nothing to Disclose: CX, LG, JZ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1163 309 2317 MON-427 PO40-01 Monday 1986 2012


1982 ENDO12L_MON-428 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) TSH, FT3 and FT4 Were Not Associated with Changes in Body Composition in HIV-Infected Patients on Combined Antiretroviral Therapy Paula Freitas, Rita Bettencourt-Silva, Ana Cristina Santos, Maria Joao Matos, Jorge Pereira, Antonio Sarmento, Jose Luis Medina, Davide Carvalho Centro Hospitalar S[atilde]o Jo[atilde]o, Faculty of Medicine, Porto, Portugal; Faculty of Medicine, Porto, Portugal; Centro Hospitalar S[atilde]o Jo[atilde]o, Porto, Portugal; Centro Hospitalar S[atilde]o Jo[atilde]o, Faculty of Medicine, Porto, Portugal [bold]Introduction: [/bold]Minor changes in thyroid function could impact on body composition of euthyroid HIV non-infected patients. The HIV-infected patients on combined antiretroviral therapy (cART) have changes in body composition, namely lipodystrophy. [bold]Aims:[/bold] To compare the levels of TSH, FT4 and FT3 in euthyroid patients with and without lipodystrophy (clinically and FMR defined) and in four groups of body composition (no lipodystrophy; isolated abdominal prominence; isolated peripheral lipoatrophy and mixed forms of lipodystrophy). [bold]Patients and methods:[/bold] Cross-sectional study of 305 HIV-1 infected patients on cART. We evaluated body composition by Dual-energy X-Ray absorptiometry (DXA), metabolic parameters and thyroid function. Fat mass ratio (FMR) was evaluated as the % of the trunk fat mass/% of the lower limbs fat mass. [bold]Results:[/bold] No differences were found between the median of TSH, FT4 and FT3 according to the presence of clinical lipodystrophy and defined by FMR and the four groups of body composition. The TSH was positively correlated with age (r=0.134; p=0.013) and negatively with CD4 cells count (r= -0.138; p=0.01). The FT4 was positively correlated with HIV infection duration (r=0.109; p=0.046) and negatively with diastolic blood pressure (DBP) (r= -0.121; p=0.026), total cholesterol (r= -0.142; p=0.009) and LDL-cholesterol (r= -0.177; p=0.001). The FT3 was positively correlated with HIV infection duration (r=0.109; p=0.048) and negatively with age (r= -0.121; p=0.027), systolic blood pressure (SBP) (r= -0.162; p=0.003) and DBP (r= -0.119; p=0.031). No statistically significant associations were observed between TSH, FT4 and FT3 and body mass index, cART duration, waist circumference, thigh circumference, glucose, HDL-cholesterol, triglycerides, leptin, adiponectin and fat mass (total, upper limbs, lower limbs and trunk). [bold]Conclusions:[/bold] No differences were observed between TSH, FT4 and FT3 values according to body composition. FT3 and FT4 were negatively associated with blood pressure unlike to that found in the HIV non-infected population.[br][br]Nothing to Disclose: PF, RB-S, ACS, MJM, JP, AS, JLM, DC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 113 309 2318 MON-428 PO40-01 Monday 1987 2012


1983 ENDO12L_MON-429 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Evaluation of Hearing Functions in Patients with Thyrotoxicosis before and after Therapy Ayse Arduc, Dilek Berker, Serhat Isik, Serpil Allusoglu, Ayse Iriz, Yasemin Tutuncu, Bercem Aycicek Dogan, Yavuz Yalcin, Serdar Guler Ministry of Health, Ankara Numune Research and Training Hospital, Ankara, Turkey; Ministry of Health, Ankara Numune Research and Training Hospital, Ankara, Turkey Introduction: Hearing functions may be affected by thyroid diseases. It is commonly known that hypothyroidism can be associated with hearing impairment. However, the relationship between thyrotoxicosis and auditory system has been investigated in a few studies. Thus we investigated hearing ability in patients with thyrotoxicosis before and after therapy.[br]Materials-Method:[br]53 normal-hearing patients with newly diagnosed thyrotoxicosis [31 Graves disease (58.5%),[bold] 9 [/bold]toxic multinodular goitre (17.0%), 13 thyroiditis (24.5%)) and 29 healthy control subjects were included in the study. Patients with histories of systemic disease, central nervous system pathology, ototoxic drug-substance use, acoustic trauma were excluded. Audiometry at 250, 500, 1000, 2000, 4000, 6000 hertz was performed in the patients and controls. The test repeated after therapy in aeuthyroid state in thyrotoxic group.[br]Results: There were no significant differences between the ages and genders of the patient and control groups (p=0.7 and p=0.329, respectively). Free triiodothyronine(FT3), free thyroxine (FT4), antithyroglobulin (anti TG) antithyroperoxidase (anti TPO), anti thyroid stimulating hormone (TSH) receptor antibody (TRAB) levels were higher, TSH level was significantly lower in the patient group (p[lt]0.05). One ear was taken into account (106 ears) hearing theresholds of thyrotoxic patients were significantly higher than controls at 250, 500, 2000, 4000, 6000 hertz. (p[lt]0, 05). A significant difference was not found between the two groups at 1000 frequency (p[gt]0, 05). Audiometric thresholds at 250, 500, 1000 frequencies showed significant decrase after the treatment (p [lt]0,05). A correlation was established between pretreatment levels of anti TPO and 250, TRAB and 250, 500,, FT3 [ndash]FT4 and 250, 500, frequencies.[br]Conclusion: Our findings revealed that hearing ability is impared in patients with thyrotoxicosis. After therapy, hearing has been improved especially at low frequencies. Additional studies are necessary for clarifying the causes of hearing impairments in thyrotoxicosis patients.[br][br]Nothing to Disclose: AA, DB, SI, SA, AI, YT, BAD, YY, SG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2318 309 2319 MON-429 PO40-01 Monday 1988 2012


1984 ENDO12L_MON-430 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Renal Tubular Creatinine Secretion Changes in Thyroid Disease Jubi E de Haan, Marinus A van den Dorpel, Arie Berghout Maasstad Ziekenhuis, Rotterdam, Netherlands [bold][italic]Introduction [/italic][/bold]Serum creatinine levels are decreased in hyperthyroidism and increased in hypothyroidism, suggesting changed glomerular filtration rate (GFR). Usually creatinine levels return to baseline after treatment. This effect may result from alterations in renal hemodynamics or a direct effect of thyroid hormone on tubular transporters as studied in animals. We hypothesize that thyroid hormone enhances renal tubular creatinine secretion (RTCS).[br][bold][italic]Methods [/italic][/bold]Patients with hyperthyroidism (TSH[lt]0.05mU/L and FT4[gt]24pmol/L) or hypothyroidism (TSH[gt]12mU/L) were eligible and were subjected to clearance studies twice: before treatment and at three months of euthyroidism. Clearance of inuline (C[sub]in[/sub]) and creatinine (C[sub]cr[/sub]) were determined after an overnight fast. C[sub]in[/sub] was determined using the continuous infusion method using sinistrin (Inutest[reg], Fresenius Kabi, Austria) and concomitant C[sub]cr[/sub] using timed urine collection. RTCS was denominated as the ratio between C[sub]cr[/sub] and C[sub]in[/sub] (C[sub]cr[/sub]/C[sub]in[/sub]).[br][bold][italic]Results [/italic][/bold]Seven patients with hyperthyroidism due to Graves[apos] disease (all females) and three patients with autoimmune hypothyroidism (2 males) were included; two hyperthyroidism patients withdrew and were excluded. In the remaining five hyperthyroid subjects TSH (0.02[plusmn]0.01 and 1.15[plusmn]1.00mU/l), FT[sub]4 [/sub](57.0[plusmn]25 and 12.2[plusmn]2.4pmol/l), total T[sub]3[/sub] (6.8[plusmn]3.0 and 1.7[plusmn]0.37pmol/l) were changed (all p[lt]0.05) before and after treatment respectively. Serum creatinine (37[plusmn]6 and 53[plusmn]8[mu]mol/l) increased and GFR (169[plusmn]21 and 147[plusmn]17ml/min/1.73m[sup]2[/sup]) decreased (NS). In the hypothyroid patients TSH (51[plusmn]17 and 3.63[plusmn]3.41mU/l), FT[sub]4[/sub] (5.9[plusmn]3.5 and 14.3[plusmn]3.6pmol/l), total T[sub]3[/sub](1.5[plusmn]0.26 and 1.7[plusmn]0.2pmol/l) changed before and after treatment. Serum creatinine (85 [plusmn]28 and 75[plusmn]20[mu]mol/l) decreased and GFR (155[plusmn]24 and 186[plusmn]39ml/min/1.73m[sup]2[/sup]) increased(NS). We observed no changes in the fractional excretion of sodium, chloride, phosphate, calcium and uric acid. After treatment of hyperthyroidism C[sub]cr[/sub]/C[sub]in[/sub] ratio fell from 2.1[plusmn]1.80 to 0.91[plusmn]0.27 (p=0.06), but after correction of hypothyroidism C[sub]cr[/sub]/C[sub]in[/sub] remained stable (0.88[plusmn]0.16 vs 0.92[plusmn]0.31. Pooled analysis of the change in C[sub]cr[/sub]/C[sub]in[/sub] ratio in both groups was near significant (1.65[plusmn]1.5 vs 0.88[plusmn]0.28;p=0.07) compared to euthyroidism.[br][bold][italic]Conclusion [/italic][/bold]We demonstrated a near significant change in RTCS in a small group of hyperthyroid and hypothyroid patients, supporting our rationale that renal tubular function might be influenced by thyroid hormone.[br][br]Nothing to Disclose: JEdH, MAvdD, AB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 680 309 2320 MON-430 PO40-01 Monday 1989 2012


1985 ENDO12L_MON-431 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Effects of Repeated Thyroid Hormone Withholding (THW) on Lipids, Weight (Wt), Waist Circumference (WCc) and High-Sensitivity Serum C-Reactive Protein (hsCRP) in Patients (pats) with Differentiated Thyroid Carcinoma (DTC) Following Total Thyroidectomy (TT) and Radioiodine Ablation (RA): Preliminary Results Doina Piciu, Andra Piciu, Robert J Marlowe, Alexandru Irimie I Chiricuta Institute of Oncology, Cluj-Napoca, Romania; Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania; Spencer-Fontayne Corporation, Jersey City, NJ; I Chiricuta Institute of Oncology, Cluj-Napoca, Romania Radically treated DTC pats traditionally undergo periodic THW to enhance follow-up testing sensitivity for disease status. THW may lead to dyslipidemia and Wt gain, but the extent and persistence of these disturbances are not well documented in modern literature. We assessed the effects of repeated THW on these variables. Since hypothyroidism has cardiovascular effects, we also examined changes in serum hsCRP, an inflammatory biomarker for cardiovascular risk.[br]Methods: We included 30 adult DTC pats (mean age 51.2 yrs, 86.6% female) given TT, RA and suppressive hormone therapy. A first THW took place from TT to RA ([quot]THW1[quot]: 4 wks); a second THW took place before the 6-month post-RA follow-up exam ([quot]THW2[quot]: 2 wks); and a 3rd THW ([quot]THW3[quot]: 2 wks) is scheduled for the 1 yr post-RA exam. We report preliminary data after two THWs. We measured serum TSH, total and HDL cholesterol (chol), apolipoprotein A1 (APL A1), triglycerides, Wt, WCc and hsCRP. Measurements occurred under euthyroidism before TT ([ldquo]baseline,[quot] [quot]BL[rdquo]), under hypothyroidism after THW1 and THW2 and under suppressive thyroid hormone doses ([ldquo]EU1,[rdquo] immediately before THW2). Our Ethics Committee approved the study; patients provided informed consent.[br]Results: At BL/THW1/EU1/THW2, mean TSH was 1.8/66.2/0.22/44.7 mIU/L (normal values, NV: 0.4-4.2 mIU/L) and mean total chol was 217/338/229/364 mg/dL (NV [lt]200 mg/dL); EU1:BL ratio was 1.05. Total chol was [gt]200-300 mg/d in 10%/40%/10%/20% of pats, [gt]300 mg/dL in 0%/60%/0%/80%. Mean HDL chol was 61.2/32.3/55.3/12.8 mg/dL; EU1:BL ratio was 0.9. HDL was beneath protective levels (60 mg/dL) in 13%/100%/0%/100% of pats. Mean triglycerides were 142.4/226.2/173/262 mg/dL (NV [lt]150 mg/dL); EU1:BL ratio was 1.21. Triglycerides were [gt]NV in 0%/96%/3%/100% of pats. Mean APL A1 was 2.03/4.2/2.41/4.9 g/L (NV: 1.04-2.25 g/L); EU1:BL ratio was 1.18. Mean hsCRP was 1.02/3.3/0.97/4.1 mg/L (low cardiac risk value [lt]1mg/L); EU1:BL ratio was 0.95. HsCRP was [gt]1 mg/L in 3%/100%/10%/100% of pats at BL/THW1/EU1/THW2. Mean Wt gain from BL to EU1 was 3.63 kg. At BL/THW1/EU1/THW2, body mass index was [gt]30 kg/m2 (obese) in 7%/10%/10%/13% of pats; mean WCc was 91.3 cm/94.9 cm/94.7 cm/98.3 cm.[br]Conclusion: Wt gain and WCc increase seemed to persist after repeated THW; THW-related dyslipidemias and elevation of the inflammatory biomarker hsCRP seemed to be transient.[br][br]Sources of Research Support: Genzyme Corporation, a Sanofi Company and the manufacturer of recombinant human TSH, supported the services of one of the authors, an independent English-language medical editor, Robert J. Marlowe, Spencer-Fontayne Corp., Jersey City, NJ, in the editorial development of this abstract.[br][br]Disclosures: DP: Advisory Group Member, Genzyme Corporation. RJM: Editor, Genzyme Corporation. Nothing to Disclose: AP, AI 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 95 309 2321 MON-431 PO40-01 Monday 1990 2012


1986 ENDO12L_MON-432 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Acute Severe Uncomplicated Hypothyroidism Is Not Associated with Hyponatremia Even with Excessive Water Intake: A Prospective Study in Thyroid Cancer Patients Fahad Almogbel, Jaber Faifi, Awad Alqahtani, Waleed Hashem, Muhammad M Hammami King Faisal Specialist Hospital [amp] Research Center, Riyadh, Saudi Arabia; King Faisal Specialist Hospital [amp] Research Center, Riyadh, Saudi Arabia; Alfaisal University, Riyadh, Saudi Arabia We have previously shown that hyponatremia is uncommon (3.9%) in severe short-term hypothyroidism in a retrospective cohort of thyroid cancer patients prepared for radioiodine therapy (1). It is not known if the nausea and stress associated with radioiodine administration together with the recommended increase in fluid intake would increase the prevalence and degree of hyponatremia.[br]We prospectively measured serum sodium levels in 171 (77% females) consecutive thyroid cancer patients before and after isolation for radioiodine treatment and studied their correlation with demographic and clinical parameters. The study was approved by the institution IRB and all patients signed an informed consent. Mean (SD) age was 40.1 (14.7) year, creatinine level 81.7 (22.0) mmol/l, TSH level 138.9 (93.4) mU/l, estimated 3-day water intake 10 (6.6) L, pre-isolation Na 140.0 (2.2) mEq/l, and post-isolation Na 138.1 (3.1) mEq/L (p[lt]0.0001 for pre and post-isolation Na, paired t test). There was no significant correlation between post-isolation Na level and TSH level (r =0.10, p =0.90) or estimated 3-day water intake (r =0.08, p =0.32). However, there was significant correlation between post-isolation Na level and age (r =-0.26, p =0.001), creatinine level (r =-0.27, p[lt]0.0001), and pre-isolation Na level (r =0.17, p =0.03). Further, the pre-post isolation drop in Na level was significantly more in females (mean difference 1.41, p =0.01). Mild hyponatremia (Na [lt]135 to [ge]130 mEq/l) was present in only 13 patients (7.6%) and moderate/severe hyponatremia (Na [lt]130 mEq/L) in 4 (2.3%). Out of the 17 patients with hyponatremia, 4 had lung metastasis, 5 were on diuretics, and 5 had elevated creatinine levels.[br]We conclude that 1) hyponatremia is uncommon in acute severe uncomplicated hypothyroidism, 2) Na level doesn[apos]t need to be routinely monitored in thyroid cancer patients undergoing radioiodine treatment unless they have lung metastasis, are on diuretics, or have elevated creatinine levels, and 3) acute hypothyroidism should not be included in the differential diagnosis of the inappropriate ADH syndrome.[br][br]1)Baajafer FS etal., J Endocrinol. Invet.1999;22:35-39.[br][br]Nothing to Disclose: FA, JF, AA, WH, MMH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1098 309 2322 MON-432 PO40-01 Monday 1991 2012


1987 ENDO12L_MON-433 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) TSH Response to Oral Glucose Tolerance Test in Patients with Obesity Duygu Yazgan Aksoy, Nese Cinar, Fatma Ucar, Mustafa Unal, Jale Karakaya, Bulent Okan Yildiz, Miyase Bayraktar Hacettepe University, Ankara, Turkey; Etlik Ihtisas Research and Training Hospital, Ankara, Turkey; Etlik Ihtisas Research and Training Hospital, Ankara, Turkey; Hacettepe University, Ankara, Turkey BACKGROUND AND AIM: Elevated TSH concentrations in association with normal or slightly elevated free T4 and/or free T3 levels have been consistently found in obese subjects, but the mechanisms underlying these thyroid hormonal changes are still unclear.Thyroid functions affect glucose metabolism. We aimed to evaluate the changes in TSH to oral glucose load in patients with obesity.[br]METHODS: Fifteen patients with the diagnosis of obesity (all female mean age 34.2[plusmn]8.3) and 16 healty age and sex matched controls were included. Age, sex, weight, body mass index (BMI), waist, fasting plasma glucose (FPG), fasting insulin (FI), lipid, thyroid stimulating hormone (TSH), free thyroxine (fT4), resting energy expenditure (REE), body composition and HOMA were measured. All patients went under 30 min oral glucose tolerance test (OGTT). Glucose, insulin and TSH were measured at 30, 60, 90 and 120. minutes. Area under curve (AUC) for glucose, insulin and TSH were calculated.[br]RESULTS: Weight, BMI, FPG, FI, TSH, REE, HOMA, basal TSH, basal glucose and insulin levels were higher in patients with obesity. After OGTT glucose and insulin levels were higher in obese group in all minutes. AUCGLU, AUCINS were higher in obese group. TSH was higher in obese groups in all minutes. AUCTSH was significantly higher in patients with obesity. TSH decreased in a stepwise fashion significantly after OGTT in both groups. Compared to basal TSH, TSH at 30, 60, 90 and 120. minutes were significantly lower in both groups. In pairwise analysis decrease in TSH was more evident in obese group. After adjusting according to basal TSH, the decrease showed similiar pattern in both groups. TSH corelated with glucose at 60 and 90 min (rs=609, p=0.016 and rs=0.880, p[lt]0.001) in obese group. AUC GLU and AUCTSH correlated also in the obese group (rs=0.538, p=0.047).[br]CONCLUSIONS: TSH is higher in patients wiht obesity. After OGTT, TSH decreased in a similar fashion in both groups. This reflects a rapid association with TSH and glucose metabolism which might result in an emphasis of thyroid gland in metabolic functions.[br][br]1. de Moura Souza A, et al., Eur J Endocrinol 2011;165:11. 2. Langer P, et al.,Endoc Reg 2000; 34:145. 3. Kok P, et al., J Clin Endocrinol Metab 2005; 90, 6185.[br][br]Nothing to Disclose: DYA, NC, FU, MU, JK, BOY, MB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1500 309 2323 MON-433 PO40-01 Monday 1992 2012


1988 ENDO12L_MON-434 POSTER SESSION: Autoimmune Thyroid Disease [amp] Thyroid Hormone Action (1:30 PM-3:30 PM) Measurement of Total Thyroxine and Triiodothyronine in Human Serum by Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry Jacob Farris, Yasamin Rahmani, Hans Cooper, Matthew Gatcombe, Julianne Cook Botelho, Hubert W Vesper Centers for Disease Control and Prevention, Atlanta, GA Triiodothyronine (T3) and thyroxine (T4) levels in blood are important for the diagnosis and treatment of hypo- and hyperthyroidism. It is estimated that more than 12 percent of the U.S. population will develop a thyroid condition during their lifetime. Although existing tests to measure total T3 and T4 are widely used in research and clinical diagnosis, inaccurate test results prevent optimal patient care and public health activates. Total serum T4 and T3 measurements are routinely performed using immunoassays, an approach with high sensitivity, but which is prone to specificity issues for many analytes. A highly specific procedure for total T3 and T4 in human serum has been developed involving isotope dilution (ID) paired with liquid chromatography tandem mass spectrometry (LC-MS/MS).[br]The method measures total endogenous T3 and T4 in serum using an automated solid phase extraction to isolate T3 and T4 form the serum matrix, followed by chromatographic separation of T3 and T4 from interfering compounds and measurement of these analytes by tandem mass spectrometry. All extraction procedures are performed in an automated manner using well plates. Internal standards (triiodothyronine-[sup]13[/sup]C[sub]6 [/sub]and thyroxine-[sup]13[/sup]C[sub]12[/sub]) are spiked into the serum material to assure accuracy and increase robustness of the assays. The measurement range of this method is 25.0- 250 ng/dL for T3 and 2.50-15.0 [micro]g/dL for T4. Measurements are performed using an API4000 mass spectrometer coupled with a TurboV electrospray ionization source and a Shimadzu HPLC system. The HPLC separation is achieved on a C18 column with a gradient of water and acetonitrile in 0.1% formic acid. Quantitation by selective reaction monitoring (SRM) of T4, T3, and internal standards is performed in the positive ion mode. A quantitation and confirmation ion peak is used for the analytes and internal standards for analysis. Method performance parameters such as sensitivity, repeatability, recovery, robustness and accuracy have been assessed.[br][br]Nothing to Disclose: JF, YR, HC, MG, JCB, HWV 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2251 309 2324 MON-434 PO40-01 Monday 1993 2012


1989 ENDO12L_MON-435 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) Thyroid Cancer Survivorship: Quality of Life, Mood and Fear of Recurrence Kristen Rasmussen, Kristin Kilbourn, Madiha Abdel-Maksoud, Joshua Klopper University of Colorado Denver, Denver, CO; University of Colorado School of Public Health, Aurora, CO; University of Colorado School of Medicine, Aurora, CO [bold]Purpose[/bold]: Research regarding the quality of life of thyroid cancer survivors is limited, and when performed often utilizes a comparison group based upon a [ldquo]normal healthy population[rdquo]. Previous studies have shown mixed results, with some thyroid cancer survivors having a worse quality of life compared to control population and others with no difference. There is even less data regarding thyroid cancer survivors[apos] fear of recurrence. Investigation surrounding fear of recurrence (FOR) in thyroid cancer survivors is important to determine ways to decrease this fear and increase psychological quality of life.[br][bold]Methods:[/bold] This is a cross-sectional study, involving an online, self-report measure examining psychological variables in thyroid cancer survivors as compared to patients with thyroidectomy but a resultant histopathology showing no cancer using the Center for Epidemiologic Studies Depression Survey. Additionally, the Fear of Cancer Recurrence scale and Functional Assessment of Cancer Therapy General (FACT-G) measures were used in the thyroid cancer cohort. This study captured patients who have had surgery in the past 10 years.[br][bold]Results:[/bold] The study enrolled 72 females (56 thyroid cancer survivors, 16 controls) and 9 males (8 cancer survivors, 1 control). The average time from surgery was 2.96 (SD=1.92) years. There was no difference in mood between thyroid cancer survivors and patients with thyroidectomy but no cancer. This was unchanged using multiple regression analysis for age, gender race or education. For thyroid cancer survivors, the average FOR score was 17.9 (SD=6.48). FOR was found to be significantly related to emotional (r = -.0484, p[lt].001), physical (r = -.0365, p[lt].01), and overall well-being (r = -.0311, p[lt].05). FOR showed no significant relationship to knowledge of disease. FOR was not related to age, time since thyroid surgery, or gender.[br][bold]Discussion:[/bold] FOR is commonly reported among cancer survivors; however, research regarding this construct is limited in thyroid cancer survivors. Thyroid cancer survivors are shown to experience FOR, which is related to a decrease in emotional, physical, and overall well being. FOR did not show a relationship to knowledge of disease in the population examined. The relationship to decreased well-being indicates that FOR needs to be further examined in thyroid cancer survivors in order to create an intervention or therapeutic solution to decrease FOR and improve overall functioning in thyroid cancer survivors.[br][br]Sources of Research Support: Cancer League of Colorado.[br][br]Nothing to Disclose: KR, KK, MA-M, JK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1859 310 2325 MON-435 PO41-02 Monday 1994 2012


1990 ENDO12L_MON-436 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) Evaluation of Quality of Life (QoL) with the ThyPRO Questionnaire in Patients with Disease-Free Differentiated Thyroid Carcinoma (DTC) Massimo Giusti, Lorenzo Mortara, Francesca Cecoli, Giorgia Pera, Valeria Caorsi, Francesco Minuto IRCCS AOUSan Martino-IST, Genoa, Italy [bold]Background [/bold]DTC patients (pts) vary widely in terms of illness perception and attention should be paid to QoL in older pts, those with recent history of disease, severe staging on diagnosis and heavier radioiodine treatment. Psychological evaluation during long-term follow-up improves QoL scores, which reach the same levels as those recorded in pts with a history of thyroid surgery for benign pathology (BP)(1) or in general population(2). A 13-scale version of the newly developed thyroid-specific ThyPRO questionnaire has recently been proposed for use in pts with BP(3). [bold]Aim [/bold]This study compared two self-rated questionnaires[ndash]the well-validated Kellner Symptom Questionnaire (KSQ) and the newly developed ThyPRO questionnaire[ndash]in terms of their ability to evaluate QoL in a cohort of cured DTC pts. [bold]Patients [/bold]We studied 110 disease-free DTC pts (age 24-88 yr; 81f, 9m) and 74 controls(age 21-88 yr; 61f, 13m) with a history of thyroid surgery for BP. Low KSQ and ThyPRO scores indicate a better QoL. [bold]Results [/bold]No difference was noted between the groups in age, male-to-female ratio, time since thyroid surgery (DTC 8.3[plusmn]0.7 yr, PB 6.8[plusmn]0.8 yr; [plusmn]SEM), use of psychotropic drugs (DTC n=18, BP n=16) or scores on the Billewicz scale (BS). As expected, significantly (p[lt]0.0001) lower TSH and higher thyroid hormone levels were found in DTC pts than in controls. No difference in the scores on the KSQ (anxiety, depression, somatisation, hostility) and ThyPRO scales (goitre, hyperthyroid, hypothyroid and eye symptoms, tiredness, cognitive impairment, anxiety, emotional susceptibility, impaired social, daily and sex life, cosmetic complaints) was found between DTC pts and controls. The mean KSQ (DTC 6.3[plusmn]0.4, BP 6.6[plusmn]0.5) and ThyPRO (DTC 19.4[plusmn]1.3, BP 20.3[plusmn]1.5) scores were similar in the two groups and a significant correlation was found between the mean scores on the two questionnaires (p[lt]0.0001) and between the anxiety scales (P[lt]0.0001). In both groups, the BS and KSQ somatisation scales were significantly (p ranging from =0.02 to [lt]0.0001) related to the ThyPRO somatic scales. No correlation was found between age and KSQ or ThyPRO scales. [bold]Conclusions [/bold]The ThyPRO can be used in DTC pts to evaluate QoL, and yields results similar to those obtained with KSQ. Several years after the diagnosis of DTC, disease-free pts presented scores similar to subjects with a history of thyroidectomy for BP. In these two groups was not documented a age-related decline in QoL by the ThyPRO questionnaire.[br][br](1)Giusti M et al.,J Zhejiang Univ Sci B. 2011; 12:163. (2)Pelttari H. et al., Clin Endocrinol (Oxf)2009; 70:493. (3)Watt T et al., Eur J Endocrinol. 2010; 162:161.[br][br]Nothing to Disclose: MG, LM, FC, GP, VC, FM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 299 310 2326 MON-436 PO41-02 Monday 1995 2012


1991 ENDO12L_MON-437 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) High Level of Distress in Long-Term Survivors with Differentiated Thyroid Carcinoma: Results of Rapid Screening Using the Distress Thermometer Sean Roerink, Mischa de Ridder, Judith Prins, Angelique Huijbers, Hans de Wilt, Henri Marres, Han Repping-Wuts, Nike Stikkelbroeck, Henri Timmers, Ad Hermus, Romana Netea-Maier Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands Background: Cancer patients are at increased risk for distress. The Distress Thermometer (DT) and problem list (PL) are short-tools validated and recommended for distress screening in oncologic patients.[br]Objective: To investigate the level of distress and problems experienced by differentiated non-medullary thyroid carcinoma (DTC) survivors, using the DT and PL and whether this correlates with clinical and demographical variables.[br]Participants and design: All 205 DTC patients, under follow-up at the outpatient clinic of our academic hospital, were asked to fill in the DT and PL, hospital anxiety and depression scale (HADS), illness cognition questionnaire (ICQ) and an ad hoc questionnaire. Receiver Operator Characteristic analysis (ROC) was used to establish the optimal cut-off score on DT according to HADS. Correlations of questionnaires scores with data on diagnosis, treatment and follow-up collected from medical records were analyzed.[br]Results: Of the 159 respondents, 145 agreed to participate (118 in remission, median follow-up 7,2 years (range 3 months-41 years)). Of these, 34.3% rated their distress score [ge]5, indicating clinically relevant distress according to ROC analysis. Patients reported physical (86,1%) over emotional problems (76%) as sources of distress. DT scores correlated with HADS scores and ICQ subscales. No significant correlations were found between DT scores and clinical or demographical characteristics except for employment status.[br]Conclusion: Prevalence of distress is high among patients with DTC even after long-term remission and cannot be predicted by clinical and demographical characteristics. DT and PL are useful screening instruments for distress in DTC patients and could easily be incorporated into daily practice.[br][br]Nothing to Disclose: SR, MdR, JP, AH, HdW, HM, HR-W, NS, HT, AH, RN-M 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1301 310 2327 MON-437 PO41-02 Monday 1996 2012


1992 ENDO12L_MON-438 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) Serologic Thyroid Antibody and Serum TSH Level as Predictors for Thyroid Malignancy in Patients with Indeterminate or Nondiagnostic Results of Fine-Needle Aspiration Biopsy Joo Hyung Kim, Sin Gon Kim, Dong Seop Choi, Sei Hyun Baik, Kyung Mook Choi, Nan Hee Kim, Ji A Seo, Hee Young Kim, Hye Jin Yoo, Sae Jung Yang, Yoon Jung Kim, Hae Yoon Choi, Chae Ryung Eun, Nam Hoon Kim, Jae Hee Ahn Korea University Hospital, Seoul, Republic of Korea Fine-needle aspiration biopsy (FNAB) is accepted as a gold standard method for evaluating thyroid nodules. However, there is a limitation of indeterminate cytologic results needing surgery. Therefore, other preoperative markers of predicting of malignancy are urgently needed. Among the markers, role of thyroid antibody and thyroid stimulating hormone (TSH) in indeterminate thyroid nodule are still uncertain. We aimed to evaluate the role of thyroid antibody and TSH level in patients with indeterminate or nondiagnostic cytology.[br]From January 2006 to December 2009, the medical records of 325 patients who had indeterminate or nondiagnostic FNAB results and subsequently underwent thyroidectomy in our hospital were retrospectively reviewed. Clinical data and untrasound (US) features was collected. TSH level, thyroglobulin antibody (TgAb) and thyroid peroxidase antibody (TPOAb) were measured. The final diagnosis as thyroid malignancy was determined by histology after surgery.[br]According to the category with indeterminate (follicular neoplasm and atypia of undetermined significance or atypical cells) and nondiagnostic cytology, malignancy rate were 16.3% (20/123), 59% (62/105) and 28% (28/97), respectively. Age and family history of thyroid cancer were similar between two groups (benign vs.malignant). Female gender and US features of small size, multiplicity and irregular margin were associated with malignant nodules (p[lt]0.05). US features of iso- or mixed echogenicity, nonexistent calcification and cystic or mixed content were associated with benign nodules (p[lt]0.01). Malignant nodules had a higher rate of positive thyroid antibodies (62% vs. 38%; p[lt]0.0001). The mean TSH level was not significantly different between two groups. But, as TSH level was higher, the percentage of malignancy was significantly increased ([lt]0.17 [micro]IU/ml, 0.17-1.17 [micro]IU/ml, 1.18-2.01 [micro]IU/ml, 2.02-4.05 [micro]IU/ml and 4.05 [micro]IU/ml[lt]; 15.8%, 30.3%, 32.8%, 40.7% and 47.1%, respectively; p=0.023). In the logistic regression analysis, positive thyroid antibody was significantly associated with thyroid cancer (odds ratio [OR] = 12.89, 95% CI 2.50-66.52, p=0.002). Our results show that thyroid antibody is an independent predictor for malignancy in thyroid nodule with indeterminate or nondiagnostic cytology, and elevated TSH level is associated with cancer risk. We suggest that thyroid antibody and TSH level can be helpful to make a decision on management of indeterminate thyroid nodule.[br][br]Nothing to Disclose: JHK, SGK, DC, SHB, KMC, NHK, JAS, HYK, HJY, SJY, YJK, HYC, CRE, NHK, JHA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 307 310 2328 MON-438 PO41-02 Monday 1997 2012


1993 ENDO12L_MON-439 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) Does Residual Thyroid Function in Hashimoto[apos]s Influence Thyroid Cancer Development? Rodis Paparodis, Kalpana Naraharisetty, Juan C Jaume School of Medicine and Veterans Affairs Medical Center, University of Wisconsin-Madison, Madison, WI [bold]Introduction:[/bold] Thyroid cancer treatment implies destruction of thyroid tissue. Hashimoto thyroiditis (HT) does just that. Hence, HT should protect against thyroid cancer. However, the relationship between HT and thyroid cancer remains controversial. We recently unraveled an association between euthyroid HT and differentiated thyroid cancer (DTC) (1). We therefore wonder if [underline]residual thyroid function[/underline] is needed for DTC to arise in patients with HT.[br][bold]Methods[/bold]: We studied how the presence of [underline]residual thyroid function[/underline] influences the development of DTC in patients with HT by analyzing data from a cohort of 56 consecutive patients who underwent thyroidectomy with pre-operative HT diagnosis. Some of these patients also had DTC, so we divided them in two groups based on the presence or absence of thyroid cancer. We first estimated the [underline]residual thyroid function[/underline] in these patients by subtracting the thyroid hormone replacement dose (as a function of TSH) from the calculated thyroid hormone requirement by weight. We then compared the [underline]residual thyroid function[/underline] in patients with HT alone to those concomitantly affected by HT and DTC.[br][bold]Results:[/bold] Out of 56 patients in our cohort, 43 were found to have HT alone, and 13 were noted to have both HT and DTC. Prior to surgery, the replacement dose in patients with HT alone, was [bold][italic]significantly[/italic][/bold] higher (lesser [underline]residual thyroid function[/underline]) compared to those with HT and DTC, 1.41 mcg/kg (range 0.19-3.29) and 1.06 mcg/kg (range 0.25-1.79) respectively. The median TSH was also higher in patients with HT alone, 2.55 uIU/mL (range 0.04-25.3), compared to the ones with HT and DTC, 1.81 uIU/mL (range 0.11-7.86). However, the disease duration was not significantly different in patients with HT alone, 4.00 years (range 0.083-29), compared to patients with both HT and DTC, 3.83 years (range 0.67-20).[br][bold]Conclusions: [/bold]HT causes T cell mediated destruction of thyroid epithelium, which in turn decreases the overall endogenous production of thyroid hormones. Once complete destruction has taken place, it is assumed that no functional thyroid epithelium remains. DTC arises from the thyroid epithelium which normally traps iodine for production of thyroid hormones. We observed [underline]residual thyroid function[/underline] in patients with HT when DTC was present. The presence of functional thyroid epithelium may explain the unexpected co-existence of HT and DTC.[br][br]1. K. Todorova-Koteva, A. Staii and J. C. Jaume*. Non-Clinical (Euthyroid) Hashimoto Thyroiditis as a Risk Factor for Thyroid Cancer. The Endocrine Society. 2010. San Diego, CA. *Presidential Award Winner.[br][br]Sources of Research Support: NIH. VA.[br][br]Nothing to Disclose: RP, KN, JCJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2140 310 2329 MON-439 PO41-02 Monday 1998 2012


1994 ENDO12L_MON-440 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) Is There an Association between Hashimoto Thyroiditis and Thyroid Cancer? Sara Ahmadi, Melinda S Moore, Daniel H Freeman, Jacob Oommen, Vicente A Resto, Judy L Ramirez, Maria D Vasquez, Randall James Urban, Ladale K St Clair University of Texas Medical Branch, Galveston, TX; University of Texas Medical Branch, Galveston, TX; University of Texas Medical Branch, Galveston, TX; University of Texas Medical Branch, Galveston, TX; University of Texas Medical Branch, Galveston, TX [bold]Background: [/bold]The association between inflammation and cancer is well established but the association between Hashimoto[apos]s thyroiditis (HT, also known as chronic lymphocytis thyroiditis) and thyroid cancer remains controversial since Lindsay first published a possible association in 1995.[br][bold]Method: [/bold]Our 12 month pilot case-control study aims to investigate the association between HT and thyroid cancer and to determine if the presence of HT is associated with a more favorable cancer staging and/or more surgical complications. Patients 18-80 years old undergoing thyroidectomy for any reason at UTMB from June 2011 forward will be enrolled. Patients with a history of radiotherapy to head or neck or radioactive iodine therapy are excluded. Informed consent is obtained and patient[apos]s blood is drawn prior to surgery to measure anti-TPO and inflammatory markers. The diagnosis of HT will be made based on histopathology finding and antiTPO level.[br][bold]Preliminary results: [/bold]Statistical analysis was performed using the Fisher[apos]s exact test and the chi-square test. A total of 15 patients (age 44+/-18, 12 Female, 3 Male) have been recruited. Preoperative hormone levels are as follows: TSH= 0.01-123 uIU/ml with SD=38.3, Free T4= 0.07-7 ng/dL with SD=1.8. Six patients (40%) were found to have thyroid cancer (4 Papillary thyroid carcinoma, 1 Medullary thyroid carcinoma and 1 Hurthle cell carcinoma). Tumor diameter in six patients with thyroid cancer was 2.7+/-2.0 cm.Four patients (26%) had HT reported in surgical path and just one had thyroid cancer (PTC). The association between HT and thyroid cancer was not statistically significant (P-Value =0.6). In six patients with cancer the presence of HT was not associated with favorable prognostic factors (vascular capsular invasion P-Value=0.2, Extra-thyroidal invasion P-Value =0.3, Lymph node metastasis P-Value 0.1). The presence of HT was not associated with more surgical complications (recurrent laryngeal nerve injury P-Value=0.3, hypocalcemia P-value=0.6). None of the patients had infection or excessive bleeding as the complication of surgery.[br][bold]Conclusion: [/bold]Preliminary results have shown no association between thyroid cancer and HT, between the presence of HT and better prognostic factors or increasing complication of surgery.[br][br]Nothing to Disclose: SA, MSM, DHF, JO, VAR, JLR, MDV, RJU, LKSC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1147 310 2330 MON-440 PO41-02 Monday 1999 2012


1995 ENDO12L_MON-441 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) Analysis of Alterations in Epigenetic Control of Gene Expression in Medullary Thyroid Carcinoma Cosimo Durante, Diego Russo, Marialuisa Sponziello, Amelie Boichard, Mariavittoria Dima, Sophie Leboulleux, Martin Schlumberger, Jean-Michel Bidart, Giuseppe Damante, Sebastiano Filetti Universit[agrave] di Roma Sapienza, Rome, Italy; Universit[agrave] di Catanzaro Magna Graecia, Catanzaro, Italy; Institut de Canc[eacute]rologie Gustave Roussy, Villejuif, France; Institut de Canc[eacute]rologie Gustave Roussy, Villejuif, France; Universit[agrave] di Udine, Udine, Italy Epigenetic control of gene expression has shown to play a major influence in the development and progression of many cancer types. Here we investigated the mRNA expression of several proteins belonging to various families involved in epigenetic control of transcription in a series of 56 medullary thyroid carcinomas (MTC). They included 13 familial MTCs, all hosting a RET mutation, and 43 sporadic forms, 20 of which with a RET somatic mutation. Gene expression profiles in cancer tissues were assessed by quantitative real-time RT-PCR using micro fluidic cards (Taqman Low Density Arrays, TLDA, Applied Biosystem) in an ABI PRISM 7900HT Sequence Detection System (Applied Biosystems). Examination of transcripts levels of Histone deacetylases 2-11, Histone acetyltransferases (HDAC) EP300, KAT3A, KAT2B, KAT2A, KAT5, MYST3, MYST4, histone methyl transferases EZH2, WHSC1, SMYD3, SMYD4, PRDM5, PRDM14, CARM1, EHMT1, PRDM2 Histone demethylases KDM1A, KDM2A, KDM3A, KDM4A, KDM5B, and DNA methyltransferases DNMT1, DNMT3A, DNMT3B revealed a light significant increase of [italic]HDAC7, KDM5B, DNMT1, 3A, 3B, SMYD3 and EHMT1[/italic] expression in the more aggressive diseases (N1,M1). The strongest increase (more than 2 fold) was observed for EZH2 gene expression. Interestingly, upregulation of this partner of polycomb group proteins has been described in a broad range of hematopoietic and solid neoplasia, associated with poor prognosis. Significant light differences were also detected in the transcripts of HDAC3, 6, 7, 8 and 10, EP300, SMYD4 and EHMT1, KAT2B, KDM1A, 2A, 4A 4C and 5B, DNMT3A and 3B genes, when comparing the RET-mutated (mainly at codon 918) with the RET-wild type tumors. In conclusion, these preliminary data demonstrate that altered expression of elements of epigenetic pathways, derived in some case by oncogenic RET activation, is associate with a more aggressive behaviour of MTCs. In particular, the histone methyltransferase EZH2 may represent a useful prognostic biomarker, as well as a new potential therapeutic target for these tumors.[br][br]Nothing to Disclose: CD, DR, MS, AB, MD, SL, MS, J-MB, GD, SF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1534 310 2331 MON-441 PO41-02 Monday 2000 2012


1996 ENDO12L_MON-442 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) The Natural History of RET Codon 790 Mutation in Medullary Thyroid Cancer: French National Study Helene Bihan, Arnaud Murat, Marinos Fysekidis, Francoise Borson-Chazot, Eric Baudin, Catherine Cardot-Bauters, Anne-Paule Gimenez-Roqueplo, Pierre-Jean Guillausseau, Jean-Marie Pochart, Isabelle Raingeard, Elisabeth Requeda, Jean-Louis Sadoul, Philippe Thieblot, Yves Resnick, Regis Cohen Avicenne Hospital, Bobigny, France; Nantes University Hospital, Nantes, France; Lyon University Hospital, Bron, France; Institute Gustave Roussy, Villejuif, France; Lille University Hospital, Lille, France; Georges Pompidou European Hospital, Paris, France; Lariboisi[egrave]re Hospital, Paris, France; Institute Jean Godinot, Reims, France; Montpellier University Hospital, Montpellier, France; Sud Francilien Hospital, Corbeuil-Essonne, France; Nice University Hospital, Nice, France; Clermont-Ferrand University Hospital, Clermont-Ferrand, France; Caen University Hospital, Caen, France [bold]Background:[/bold] Mutations of the [italic]RET[/italic] protooncogene are responsible for Multiple Endocrine Neoplasia (MEN) type 2. The strong genotype-phenotype correlation and the aggressiveness of the tumor related mutation dictates the decision for the age of thyroidectomy, but in the absence of adequate evidence on less common mutations stricter guidelines for prophylactic thyroidectomy may be applied as a mean of precaution. [br][bold]Patients and Methods:[/bold] We report 78 carriers of the 790 codon intracellular [italic]RET[/italic] gene mutation from 13 french centers and clustered data of 19 families. [br][bold]Results:[/bold] From 85 mutation carriers, 7 were excluded for insufficient data. The average age at diagnosis was 36.1 years [plusmn] 20.2, and 39 were women. Fifty three patients had thyroidectomy for medullary cancer at the age of 5 to 82 years, with an average age of 38 years. TNM staging was as follows: T0N0M0 in 19 patients, T1N0M0 in 24, T1N1M0 in 6, T2N1M0 in 2, T3N1M0 in 2. Patients with no adenopathy were younger than those operated and classified as N1 (mean age 33 (range 5 to 76) vs. 50 (32 to 82) years, p[lt] 0.004). In the N0 group, 84% were considered cured after surgery (42/52) with a mean follow up of 6.8 years based on the calcitonin levels; whereas in the N1 group, 5 had multifocal disease and 3 out 9 were cured.[br]Twenty five patients did not have surgery (ages 1.5 to 78 years): 12 patients had no follow up, 11 had normal basal calcitonin levels out of the other 13. The other 2 patients with an abnormal calcitonin levels had a mean follow up of 3.1 years and the surgery was delayed. Age and gender were not significant predictors for remission. Only one patient had phaeochromocytoma.[br][bold]Conclusion:[/bold] Among the carriers of a mutation in codon 790 of the RET gene, about 1/3 had no thyroidectomy, 1/3 had surgery without tumor evidenced, and 1/3 underwent thyroidectomy with discovery of a tumor. The absence of distant metastases and the slow evolution in unoperated patients are in favour of a less aggressive form of RET mutation. We suggest that change in calcitonin levels or procalcitonin assessment could be interesting topics to precise in future studies regarding the aggressiveness of this mutation.[br][br]Nothing to Disclose: HB, AM, MF, FB-C, EB, CC-B, A-PG-R, P-JG, J-MP, IR, ER, J-LS, PT, YR, RC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1265 310 2332 MON-442 PO41-02 Monday 2001 2012


1997 ENDO12L_MON-443 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) Barriers to Genetic Testing of Families with Inherited Cancer Syndromes: Example of Inherited Medullary Thyroid Carcinoma Aysha E Inankur, M Sara Rosenthal, Clifton Iler, Kristy S Deep, Anna V Marino, Kenneth B Ain University of Kentucky, Lexington, KY; Veterans Affairs Medical Center, Lexington, KY; University of Kentucky, Lexington, KY; University of Kentucky Medical Center, Lexington, KY Introduction: In familial cancer syndromes with defined germline genetic etiologies, identification of an index case should lead to family screening for further case-finding and early treatment. In familial medullary thyroid cancers (FMTC), associated with a mutation of the RET proto-oncogene, such screening has become standard-of-care (1) and is ethically mandated (2). The relative ease of prophylactic thyroidectomy in affected family members, contrasted to mastectomy and oophorectomy with BRCA mutations or colectomy in inherited polyposis syndromes, suggests that genetic screening should be easily accepted for FMTC; however, uptake of this testing is far less than 100%. This suggests significant barriers to testing. We present qualitative data regarding barriers to RET mutation screening in an at-risk Appalachian kindred.[br]Methods: Semi-structured phone [amp] face-to-face interviews (IRB approved [amp] HIPAA consent obtained from index patient) were conducted with 23 family members and 37 letters sent. Investigation identified 75 at-risk members, of which 5 were already deceased. Letters were written at 8[sup]th[/sup] grade reading level, with enclosures for distribution to relatives, and follow-up phone calls made.[br]Results: Barriers were encountered 47 times during structured interviews. Major emergent themes included: interpersonal family conflict, screening cost, inadequate insight, inadequate comprehension and distrust of the medical team. Over 16 months after initiating screening, only 22 individuals had been tested, 10 had declined testing, and 8 could not be contacted directly. Thirteen members tested positive while 37 remained at risk. Nearly 40 hours of unreimbursed physician and staff time were required in these efforts to notify at-risk family members.[br]Conclusions: Physicians face multiple barriers to effect genetic screening in order to fulfill their duty to warn and deliver healthcare to kindreds with genetic cancers. Cost of testing, educational level, and communication within the family influences whether members comply with testing. The family[apos]s social and cultural landscape affects barriers to screening. Despite significant unreimbursed time and effort, a majority of at-risk members remain unscreened. This presents a public health and ethical dilemma possibly requiring involvement of public health resources, as has been done for tuberculosis control (3). Alternatively, it could prove more cost-effective to implement national genetic testing of newborns (4).[br][br](1) Kloos RT, Eng C, Evans DB, Francis GL, Gagel RF, Gharib H, Moley JF, Pacini F, Ringel MD, Schlumberger M, Wells SA, Jr. 2009 Medullary thyroid cancer: management guidelines of the American Thyroid Association. Thyroid 19:565-612. (2) Rosenthal MS, Pierce HH 2005 Inherited medullary thyroid cancer and the duty to warn: revisiting Pate v. Threlkel in light of HIPAA. Thyroid 15:140-145. (3) Sepkowitz KA 1994 Tuberculosis and the health care worker: a historical perspective. Ann Intern Med 120:71-79. (4) Rosenthal MS, Diekema DS 2011 Pediatric ethics guidelines for hereditary medullary thyroid cancer. Int J Pediatr Endocrinol 2011:847603.[br][br]Nothing to Disclose: AEI, MSR, CI, KSD, AVM, KBA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1657 310 2333 MON-443 PO41-02 Monday 2002 2012


1998 ENDO12L_MON-444 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) Comparison of Real-Time, High-Resolution Ultrasonography of the Vocal Folds with Videolaryngostroboscopy in Patients before and after Thyroid Surgery Marek Dedecjus, Zbigniew Adamczewski, Jan Brzezinski, Marek Ruchala, Andrzej Lewinski Medical University of Lodz, Polish Mother[apos]s Memorial Hospital - Research Institute, Lodz, Poland; Medical University of Lodz, Polish Mother[apos]s Memorial Hospital - Research Institute, Lodz, Poland; Poznan University of Medical Sciences, Poznan, Poland Purpose: The aim of the study was to evaluate the functionality of vocal folds by real-time, high-resolution ultrasonography and to compare the results with a golden standard in vocal folds imaging [ndash] videolaryngostroboscopy.[br]Patients and Methods: The study group comprised 82 patients (60 females and 22 males),qualified to thyroidectomy. All the patients had the ultrasound examination and videolaryngostroboscopy performed before, two days after, and six months after the thyroid operation. We used high resolution US-imaging to identify vocal folds, and subsequently a pulsed Doppler and Doppler gate to quantify the tissue displacement velocity in the vibrating vocal fold section.[br]Results: Videolaryngostroboscopy revealed unilateral vocal fold paralysis in eight patients. Vocal fold dysfunction was diagnosed in other twelve subjects. In simultaneously performed US-examination, changes in vocal fold displacement velocity (VFDV) were observed in twenty five patients. In eight patients subjects, VFDV was below 30 cm/s [ndash] the patients with vocal fold paralysis, diagnosed in videolaryngostroboscopy. In further 17 cases, we observed VFDV decrease by 50%, comparing to preoperative values. Both US-imaging and videolaryngostroboscopy examination, performed after the three month follow-up, confirmed the transitory character of most of the above-mentioned pathologies.[br]Conclusions: US-imaging of the vocal folds correlated with videolaryngostroboscopy results, while being a minimally invasive, easily reproducible and inexpensive method of examining vocal folds functionality. Thanks to many recording options, it may become a perfect tool for an early identification of postoperative vocal folds dysfunction with its later monitoring in patients after thyroid surgery.[br][br]Nothing to Disclose: MD, ZA, JB, MR, AL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1532 310 2334 MON-444 PO41-02 Monday 2003 2012


1999 ENDO12L_MON-445 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) Vestigial Thyroid Tissue Identified in Pre-Treatment Iodine-123 Whole-Body Scans in Patients with Follicular-Derived Thyroid Cancer: Diagnosis and Management of Lingual Thyroid and Thyroglossal Duct Remnants Kerri A Kissel, Craig S Cochran, James Reynolds, Francesco S Celi National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD; National Institutes of Health, Bethesda, MD Lingual thyroid and thyroglossal ducts are congenital anomalies that result from the persistence of thyroid tissue past embryologic migration of the primodordial thyroid from the neotongue, passing anterior to the hyoid bone on its path to the final anatomical position of the thyroid gland. Thyroid remnant ablation by Iodine 131 ([sup]131[/sup]I) is one mainstay of therapy for Follicular-Derived Thyroid Cancer (FDTC). Vestige thyroid tissue along the thyroglossal duct are rarely identified during diagnostic or post-therapy Nuclear Medicine scans. Here we present two cases that illustrate two rare thyroid related congenital anomalies and their clinical relevance in the management of FDTC.[br]Case 1: A 25 year old female with multinodular goiter underwent fine needle aspiration biopsy of the dominant nodule in the left thyroid lobe. Cytology demonstrated architectural and focal atypia. Subsequently, the patient underwent thyroidectomy. Final diagnosis was consistent with papillary thyroid cancer with pretracheal lymph node metastasis. No vascular invasion was noted. The patient underwent human recombinant TSH-stimulated pretreatment [sup]123[/sup]I whole body scan that revealed two focal areas of uptake in the cervical region cephalic to the thyroid cartilage, consistent with thyroglossal duct. This was confirmed with SPECT image reconstruction and CT. The patient underwent uncomplicated treatment with 150 mCi [sup]131[/sup]I.[br]Case 2: A 54 year old male with history of stage III, T3NxMx PTC underwent pretreatment [sup]123[/sup]I whole body scan while on thyroid hormone withdrawal. The scan demonstrated an intense focus in the upper cervical region. The area of intense uptake was located at the level of the chin in the oropharynx. SPECT image reconstruction and CT localized the area of uptake to the base of the tongue, adjacent to the epiglottis, consistent with lingual thyroid without evidence of thyroglossal duct cyst. The patient was initially treated with a low dose 10 mCi [sup]131[/sup]I to ablate the lingual thyroid to avoid airway compromise before undergoing high-dose [sup]131[/sup]I treatment. The patient tolerated the low dose of [sup]131[/sup]I well without airway compromise.[br]The cases contrast the varying clinical presentations of vestigial thyroid residues along the thyroglossal duct during management of FDTC. Of note, the discovery of a lingual thyroid and its ablation with low-dose [sup]131[/sup]I likely prevented airway complications due to acute radiation-induced inflammation and edema of the lingual thyroid adjacent to the epiglottis.[br][br]Sources of Research Support: This work was supported by the Intramural Research Programs of NIDDK and NICHD.[br][br]Nothing to Disclose: KAK, CSC, JR, FSC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2185 310 2335 MON-445 PO41-02 Monday 2004 2012


2000 ENDO12L_MON-446 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) Metastasis to the Thyroid Gland: Report of a Large Series from the Mayo Clinic, Rochester Livia Hegerova, Marcio L Griebeler, Jordan Reynolds, Michael Henry, Hossein Gharib Mayo Clinic Rochester, Rochester, MN; Mayo Clinic Rochester, Rochester, MN; Mayo Clinic Rochester, Rochester, MN; Cleveland Clinic, Cleveland, OH Introduction: Metastases to the thyroid gland are not as unusual as previously believed. Autopsy series have demonstrated metastases to be more common than primary thyroid malignancies. We report our experience with a large group of patients with thyroid metastasis, confirm the accuracy of fine needle aspiration (FNA) in diagnosing metastasis, and identify the most common primary tumors in the thyroid gland.[br]Methods: This study reviews all thyroid FNAs performed at Mayo Clinic-Rochester from 1980-2010. We identified 97 patients with a metastatic solid neoplasm of the thyroid gland. Retrospective analysis included the frequency and types of malignant lesions, clinical course and survival after thyroid involvement. Statistical analysis was performed using Kaplan-Meier survival estimates.[br]Results: Frequent primary tumor sites included kidney (23%), lung (22%), head and neck (12%), breast (11%), esophagus (9%), skin (7%), neuro (5%) and uterus (3%). The most common tumor types were adenocarcinoma and squamous cell carcinoma. FNA cytology detected metastatic malignancy in 91 of 97 patients. The sensitivity for detecting metastases was 94%, and the specificity was 100%. Six cases were diagnosed incorrectly as follicular neoplasm or papillary thyroid carcinoma after FNA. Two patients had thyroid involvement before a primary malignancy was identified.[br]The average age at primary diagnosis was 59; average age at the time of discovery of thyroid metastasis was 63. The time from diagnosis of primary tumor to metastasis to the thyroid gland was most considerable for renal cell carcinoma (mean 113 months). 41 patients underwent thyroid resection with an average metastatic tumor size of 3 cm. Median survival in all patients with metastases was 20 months (1-228 months). Patients who underwent thyroid resection had a median survival of 30 months (3-171 months) while survival in patients without thyroid surgery was 12 months (1-228 months, p=0.09). 25% of patients were alive at 5-years after metastases.[br]Conclusions: Our experience over the last 30 years confirms that FNA remains a sensitive and specific method to detect metastases to the thyroid. The most common primary origin of metastases, in order, was from kidney, lung, and head and neck. In any patient with a history of a malignancy, a new thyroid mass should be promptly evaluated for recurrent malignancy as early diagnosis and surgical resection resulted in a non-statistically significant increased median survival.[br][br]Nothing to Disclose: LH, MLG, JR, MH, HG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 711 310 2336 MON-446 PO41-02 Monday 2005 2012


2001 ENDO12L_MON-447 POSTER SESSION: Thyroid Cancer: Survivorship Issues [amp] Management (1:30 PM-3:30 PM) A Non-Surgical Approach to the Treatment of Metastatic Follicular Thyroid Cancer (FTC) in a Medically High-Risk Patient Jennifer Zaitz, Norman H Ertel, Maya Raghuwanshi, John J Shin UMDNJ-New Jersey Medical School, Newark, NJ; Veterans Affairs, East Orange, NJ [bold]Introduction: [/bold][br]Follicular thyroid cancer is treated with total thyroidectomy followed by radioactive iodine (RAI) ablation of residual thyroid tissue post operatively. We have not identified published accounts of RAI thyroid ablation as a primary treatment for metastatic FTC in a patient who is not a surgical candidate; we present such a case.[br][bold]Case: [/bold]An 84-yr-old male with a history of COPD, requiring home O2, underwent a CT scan of his chest as part of an evaluation of a lung nodule and was found to have thyroid nodules. These nodules, evaluated by ultrasound and nuclear imaging, measured 2.3x2.6cm and 3.1x2.1cm in size and were photopenic on thyroid scan. The patient underwent FNA; the cytology was suspicious for follicular neoplasm. The patient was evaluated as high risk for surgery and no further action was taken.[br]Six months later, a CT scan of his chest showed an enlarging soft tissue mass posterior to the right kidney and the lung nodules had increased in size. The retro-renal mass was biopsied and identified as metastatic FTC. An I-131 scan showed uptake in the thyroid, left chest, right shoulder and posterior to the right kidney. A 25mCi dose of RAI was given since he could not undergo surgery.[br]Two months later the patient presented with severe hip pain. An MRI showed additional metastatic lesions to the thoracic spine, ileum and acetabulum; he was treated with 3,000 cGy of external radiation with symptom relief until six months later, when new pain developed in his thoracic spine. Higher dose RAI as a palliative measure for pain and prevention of spinal cord compression was considered. In preparation, the patient was treated with prednisone to decrease the likelihood of thyrotoxicosis and lithium to concentrate the RAI within thyroid tissue. Following treatment with an additional 200mCi his pain resolved and he was started on levothyroxine therapy to suppress his TSH to [lt] 1. At one year follow-up, he is pain free.[br][bold]Conclusion: [/bold][br]High dose RAI ablation may be a useful palliative treatment in thyroid cancer in patients in whom surgery is too high risk. In our patient with stage IVB FTC, his five year survival rate is 50%. He was at high risk for cord compression as a result of metastatic disease in his spine that could worsen his quality of life and shorten his survival. Since this patient had a remarkable reduction in pain and is currently thriving, primary palliative thyroid ablation may have role in similar patients.[br][br]Sampson, E., Brierley, J. D., Le, L. W., Rotstein, L. and Tsang, R. W. (2007), Clinical management and outcome of papillary and follicular (differentiated) thyroid cancer presenting with distant metastasis at diagnosis. Cancer, 110: 1451[ndash]1456. doi: 10.1002/cncr.22956. Kitamura, Yutaka et al. (1999) Immediate Causes of Death in Thyroid Carcinoma: Clinicopathological Analysis of 161 Fatal Cases. JCEM, 84: 4043-4049.[br][br]Nothing to Disclose: JZ, NHE, MR, JJS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1769 310 2337 MON-447 PO41-02 Monday 2006 2012


2002 ENDO12L_MON-449 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Treatment of Cushing Syndrome with Mifepristone: Impact over Time on Insulin Sensitivity Amisha Wallia, Kathleen Colleran, Coleman Gross, Mark Molitch Feinberg School of Medicine, Northwestern University, Chicago, IL; University of New Mexico Health Sciences Center, Albuquerque, NM; Corcept Therapeutics, Menlo Park, CA [bold]Hypothesis:[/bold] Our goal was to examine whether the improvement in insulin sensitivity seen with mifepristone (glucocorticoid receptor antagonist) is independent of changes in body weight (WT) and waist circumference (WC). [bold]Methods:[/bold] Adults with Cushing[apos]s syndrome, who completed SEISMIC (a study examining the safety and efficacy of mifepristone) with hypertension alone (C-HT) (n=11) or with diabetes/impaired glucose tolerance (C-DM) (n=17) [amp] who had evaluable data at all major study efficacy visits (wks 6, 10, 16, [amp] 24) were included in these analyses. WT, WC, fasting insulin [amp] glucose (GLU) were measured, [amp] 75g 2 hr oral GLU tolerance tests (oGTT) with insulin levels were performed. Insulin sensitivity (Matsuda Index, MI) was calculated for all C-DM not on insulin (n=9) [amp] C-HT (n=11). Statistical analysis included paired t-tests and linear mixed models for continuous variables. Data are shown as mean [plusmn] SEM. [bold]Results:[/bold] From baseline to wk 6 in C-DM, there was a -1.69[plusmn]0.81% (p=0.09) change in WT, a -0.74[plusmn]1.02% (p=0.42) change in WC, [amp] a marked change of -19.49[plusmn]4.59% (p=0.001) in GLU Area Under the Curve (AUC) on oGTT. From wk 6 to 24, there were marked changes in WT (-7.11[plusmn]1.77%, p=0.014) and WC (-8.01[plusmn]1.20%, p=0.003) with a relatively smaller change in GLU-AUC (-14.6%[plusmn]6.01%, p=0.550). Regression analysis confirmed a nearly identical continuous linear best fit model for both changes in WT (r=-0.998, p=0.0001) [amp] WC (r=-0.981, p=0.003); a two-phase model (0-6wks, [amp] 6-24wks) was a much better fit than a linear model (p=0.005 for model difference) for change in GLU-AUC, suggesting a biphasic effect on AUC with the first phase being independent of changes in WT or WC. In C-HT [amp] C-DM not on insulin, further evaluation of insulin sensitivity (MI) over time showed that more than 2/3 of the improvement in MI occurred between wk 0 [amp] wk 6 (64.92%[plusmn]21.96, p=0.018) as compared to an non-significant change from wk 6 to 24 (31.33%[plusmn]9.86, p=0.15). Concurrently, at wk 6 there were minimal changes in WT (-1.16%[plusmn]0.69%, p=0.11) [amp] WC (-1.19%[plusmn]0.74% p=0.08) as compared to significant changes seen between wks 6 [amp] 24 (WT -6.69%[plusmn]1.42%, p=0.002) (WC -6.41%[plusmn]1.27%, p=0.006). [bold]Conclusion:[/bold] Mifepristone use was associated with an early improvement in insulin sensitivity prior to any significant change in WT or WC, likely due to direct glucocorticoid receptor blockade. Later in the course of treatment, the change in WT and WC had additional effects in increasing insulin sensitivity.[br][br]Disclosures: AW: Coinvestigator, Corcept Therapeutics. KC: Coinvestigator, Corcept Therapeutics. CG: Employee, Corcept Therapeutics. MM: Consultant, Corcept Therapeutics; Investigator, Corcept Therapeutics. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1016 311 2338 MON-449 PO02-01 Monday 2007 2012


2003 ENDO12L_MON-450 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Predicting and Managing Hypertension and Hypokalemia in Cushing Syndrome during Mifepristone Therapy Richard J Auchus, Michael J McPhaul, James W Findling, Coleman Gross University of Michigan, Ann Arbor, MI; University of Texas Southwestern Medical Center, Dallas, TX; Medical College of Wisconsin, Milwaukee, WI; Corcept Therapeutics, Menlo Park, CA Hypertension and hypokalemia are frequent manifestations of Cushing[apos]s syndrome (CS), because cortisol activates both the glucocorticoid receptor (GR) and the mineralocorticoid receptor (MR). We analyzed blood pressure (BP) and potassium (K) data from the SEISMIC trial of 50 CS patients treated with the GR antagonist mifepristone (MIFE, 300-1200 mg/d) to determine prevalence, predictive factors, and approaches to management.[br]At baseline, 5 and 18 patients required spironolactone or K supplements, respectively, to maintain normokalemia, as specified by the protocol. During the trial, spironolactone was administered to 14 patients (25-400 mg/d) and K supplements to 33 (10-420 mEq/d) for hypokalemia. One or more episodes of hypokalemia were noted in 20 participants, 16 of whom were normokalemic at final study visit (80%). Those who required treatment for K wasting had higher urinary free cortisol (UFC) values at baseline as well at peak (week 10) than those who did not.[br][underline]Baseline[/underline][br]Mean[plusmn]SD = 1371[plusmn]3523 vs 317[plusmn]228 nmol/24h [497[plusmn]1277 vs 115[plusmn]83 [micro]g/24h][br]Median [range, IQR] = 289 [59-19739, 155-1017] vs 266 [85-1032, 229-324] nmol/24h[br][underline]Wk 10[/underline][br]Mean[plusmn]SD = 3668[plusmn]6094 vs 1103[plusmn]1003 nmol/24h [1330[plusmn]2209 vs 400[plusmn]364 [micro]g/24h][br]Median [range, IQR] = 938 [41-23480, 286-4417] vs 749 [32-2630, 312-2131] nmol/24h[br](p=NS for all)[br]Patients who required K treatment prior to starting MIFE had higher UFCs than those treated after starting MIFE (mean[plusmn]SD = 2194[plusmn]4654 vs 383[plusmn]387 nmol/24h [795[plusmn]1687 vs 139[plusmn]140 [micro]g/24h], p=0.03). Baseline UFC was [gt]500 nmol/24h (180 [micro]g/24h) in 14/35 participants who required treatment for hypokalemia but only 2/17 participants who did not.[br]Systolic BP (SBP) rose during the trial in 18 participants, 10 of whom had clinical evidence of mineralocorticoid excess. Overall, the mean change in SBP was -2.5 mmHg during the trial (p=NS) in the 40 patients with hypertension, but changes in antihypertensive therapies among 19 diabetic subjects confounded analysis of predictive factors. In repeated measures mixed models, UFC was positively correlated with SBP and negatively correlated with serum K (p = 0.02 for both).[br]We conclude that hypertension and hypokalemia are common in CS patients requiring GR antagonist therapy with UFC [gt]500 nmol/24h. These patients require concomitant treatment with a MR antagonist and/or K supplements, which control BP and K in the majority of cases. Clinical findings of MR activation are less predictable in patients with UFC [lt]500 nmol/24h.[br][br]Sources of Research Support: Corcept Therapeutics.[br][br]Disclosures: RJA: Investigator, Corcept Therapeutics, Novartis Pharmaceuticals. MJM: Investigator, Corcept Therapeutics, Novartis Pharmaceuticals. JWF: Investigator, Corcept Therapeutics, Novartis Pharmaceuticals; Consultant, Corcept Therapeutics, Novartis Nutrition, Inc. CG: Employee, Corcept Therapeutics. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 965 311 2339 MON-450 PO02-01 Monday 2008 2012


2004 ENDO12L_MON-451 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Receptor Activator of Nuclear Factor Kappa-B Ligand (RANKL) and Osteoprotegerin (OPG) Levels in Patients with Cushing Syndrome Zhanna Evgenjevna Belaya, Natalja Vjacheslavovna Dragunova, Alexander Victorovich Iljin, Liudmila Yakovlevna Rozhinskaya, Galina Afanasjevna Melnichenko The National Research Center for Endocrinology, Moscow, Russian Federation Osteoclasts are of monicytes origin. Autoimmune disease, to treat which glucocorticods (GC) are frequently prescribed for, might contribute to the stimulation of bone resorbtion which is followed by a profound decrease in bone formation in patients with exogenous hypercortisolism. In patients with endogenous Cushing[apos]s syndrome (CS), we can investigate the effects of excess GC on bone metabolism, minimizing other confounding factors. The goal of the study was to analyze the influence of CS on the key molecules of osteoclasogenesis (sRANKL, OPG).[br]Materials and methods: From 40 patients with active CS and 40 healthy matched individuals, fasting blood samples were taken and analyzed on the markers of bone remodeling (osteocalcin (OK), C-telopeptide fragments of type-1 collagen- (sCTx)) with automated ECLIA (Cobas e601) and sRANKL, OPG assayed by ELISA. The level of urinary free cortisol (24h UFC) was measured in all the patients by an immunochemiluminescence assay (extraction with diethyl ether) on Vitros ECi. The participants were questioned on their recent low traumatic fractures and then a thoracic and lumbar X-Ray was performed to reveal any vertebral fractures. The bone mineral density (BMD) was measured by DXA (Lunar) in patients with CS.[br]Results: The patients with CS (Me-30 (Q25 [ndash]Q75: 26-40) years old) did not differ from the healthy control (28 (24-38)) in age p=0.34, body mass index p=0.24 or sex (77,5% females). None of the healthy individuals, but 18 (45%) of the patients with active CS (24h UFC [ndash] 2575 (1184-4228) nmol/l), had low traumatic fractures with a mild reduction of BMD Z-score Neck -1,2 (-0,8- -1,9); L1-L4 -2,1 (-1 - -2,6).OC level was significantly lower in patients with CS (p[lt]0.0001), sCTx did not differ from the healthy control (p=0.59). RANKL level was significantly suppressed in patients with CS (0,08 (0,07-0,09)) pmol/l vs healthy individuals 0,1 (0,08-0,13) pmol/l p[lt]0.001 but OPG level did not differ (p=0.14). The OC and sRANKL levels significantly correlated with 24 h UFC R= -0.47 and R= -0.45 respectively (p[lt]0.05). Patients with CS and fractures had a significantly higher level of 24hUFC (p=0.05) and a lower level of sRANKL (p=0,045) and OC (p=0.019) vs patients without fractures.[br]Conclusions: Patients with CS have a profound suppression of sRANKL and OC that might reflect the lack of osteoblasts and their activity, whereas sCTx remains unchanged because of some different signals evoking bone resorption activity in patients with CS.[br][br]Sources of Research Support: The Presidential Grant No. MK-6978.201.[br][br]Nothing to Disclose: ZEB, NVD, AVI, LYR, GAM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1114 311 2340 MON-451 PO02-01 Monday 2009 2012


2005 ENDO12L_MON-452 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Hypercoagulable State in Cushing Syndrome Is Reversible after Remission Achievement Tina Dusek, Ivana Kraljevic, Izet Aganovic, Darko Kastelan Zagreb University School of Medicine, Zagreb, Croatia Hypercoagulability is a commonly described complication in patients with Cushing[apos]s syndrome (CS). Increased risk of thromboembolism has been reported both in patients with endogenous cortisol hypersecretion and in those treated with glucocorticoids. Recent clinical studies have indicated various abnormalities of different coagulation and fibrinolysis parameters which may be responsible for that phenomenon. It is still unknown whether and for how long the risk of thromboembolism is increased after the remission of CS. The issue is important in order to determine the appropriate duration of thromboprophylaxis. The present study was designed to assess the long-term effects of surgical remission of CS on coagulation and fibrinolytic markers. Eighteen consecutive patients with active CS (16 women, 2 men; age 38.6[plusmn]13.7 yr) were enrolled in the study. The control group comprised 18 age and sex matched healthy individuals (14 women, 4 men; age 40.3[plusmn]13.6 yr). Fifteen patients had corticotroph pituitary adenoma and three had cortisol producing adrenal adenoma. Coagulation and fibrinolysis markers were measured at enrolment (1-30 days before the surgery) and 6 months after achievement of disease remission. Six months after remission achievement, the CS patients had lower levels of factors II (P [lt] 0.001), V (P = 0.02), XI (P = 0.04) and XII (P [lt] 0.001), protein C (P [lt] 0.001), protein S (P = 0.001), antithrombin (P = 0.03), antithrombin Ag (P = 0.008), plasminogen (P = 0.03) and C1 inhibitor (P = 0.001), and consequently the aPTT values were shorter than at enrolment (P = 0.001). Levels of PAI-1 and factors VII, VIII and IX tended to normalize, but the differences in those parameters were not significant between the two measurements. No differences were found in haemostatic and fibrinolytic markers between the patients in remission and the control group, except for the factor XII (P = 0.02) and protein C (P = 0.004) which were lower in the remission group. We conclude that six months after achievement of CS remission, the risk of thromboembolism is the same as in the normal population.[br][br]Nothing to Disclose: TD, IK, IA, DK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1112 311 2341 MON-452 PO02-01 Monday 2010 2012


2006 ENDO12L_MON-453 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Screening for Cushing Syndrome in Overweight, Type 2 Diabetic Hospitalized Patients in the Absence of Specific Signs or Symptoms of Hypercortisolism Natalia Volkova, Maria Antonenko, Lilia Ganenko Rostov State Medical University, Rostov on Don, Russian Federation; Municipal Hospital No 4, Rostov on Don, Russian Federation Objective: diagnosis of Cushing[apos]s Syndrome (CS) is primarily based on the signs and symptoms of the hypercortisolism. The data of recent studies showed a huge number of patients with CS displayed only obesity or type 2 diabetes mellitus (T2DM). The aim of the study has been to determinate the prevalence of CS in overweight inpatients with T2DM devoid of specific clinical signs of hypercortisolism. Methods: 133 overweight inpatients with T2DM were studied. Noone had evidence of clinical hypercortisolism. A first screening test was the 1-mg overnight dexamethasone suppression test (DST) (cutoff 50nmol/l). A second confirmatory step was performed in patients with impaired 1-mg DST and included midnight plasma cortisol concentration ([lt]207nmol/l), plasma cortisol circadian rhythm ([lt]50), 24-h urinary free cortisol ([lt]180mcg/24h). Patients with impaired 1-mg DST with at least one abnormal confirmatory test result passed to the third step. The goal was to determine the source of hypercortisolism. It included morning plasma ACTH concentration (5-46pg/ml), 8-mg DST (autonomous secretion [gt]27,7nmol/l) and imaging studies. Results: among 133 patients, 22 had impaired 1-mg DST. Among 22, 4 patients droped of the study and 18 were further evaluated. 4 patients were considered as false positives of the 1-mg DST. 14 exhibited at least one additional abnormal result of the second step tests. 1 patient refused next investigations. Definitive CS was diagnosed in 2 patients (pituitary microadenoma, 1,5 % of the whole series). Definitive diagnosis remains to be established in 2 patients. The first one had bilateral adrenal adenomas with autonomous cortisol secretion without ACTH supression. The second one had biochemical signs of ACTH-dependent CS (probably, pituitary origin according 8-mg DST test result) but nothing was found on pituitary MRI. 9 patients had autonomous cortisol secretion according test results of 2, 3 steps, but nothing was found on imaging studies. The repeated tests results in 3 month didn[apos]t show autonomous cortisol secretion in 5 patients, remaining 4 patients are under investigation now. Conclusion: a relatively high prevalence of CS in our study argued, that establishing diagnosis of CS on specific clinical features, other patients with mild CS are out of sight. Probably, further studies are needed to evalute biochemical differences between normal subjects and CS patients, and, based on potential findings, to work out a rationale systematic screening.[br][br]Nothing to Disclose: NV, MA, LG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1893 311 2342 MON-453 PO02-01 Monday 2011 2012


2007 ENDO12L_MON-454 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) The Role of a Glucocorticoid Receptor [beta] SNP in Human Disease Christine M Jewell, Lisa Murphy, Stavros Garantziotis, John A Cidlowski NIEHS/National Institutes of Health, Research Triangle Park, NC; SRA International Inc, Durham, NC; NIEHS/National Institutes of Health, Research Triangle Park, NC Glucocorticoids are important mediators in the pathogenesis of cardiovascular disease and metabolic syndrome and exert their effects through glucocorticoid receptors (GR), which are expressed in most cells of the human body and regulate the expression of multiple downstream genes. Alternative splicing of the hGR gene (NR3C1) results in two isoforms; hGR[alpha] which is the dominant, active form and hGR[beta] which is a dominant negative inhibitor of hGR[alpha]. Mutations in both hGR[alpha] and hGR[beta] can cause an altered response to glucocorticoids. A single nucleotide polymorphism (SNP) in the 3[apos]UTR of hGR[beta] (hGR9[beta]A3669G) changes a destabilizing ATTTA motif to GTTTA and [italic]i[/italic][italic]n vitro[/italic] assays have demonstrated that this mutation stabilizes hGR[beta] leading to increased hGR[beta] levels. This increase in hGR[beta] may contribute to the observed phenotype of glucocorticoid resistance and the increased risk of adverse cardiovascular events in Caucasian populations. However, the incidence of the hGR9[beta] SNP and other GR SNPs has not been determined in non-Caucasian populations. Our study investigates the functional relevance of the hGR9[beta] SNP in individuals from the NIEHS Environmental Polymorphism Registry (EPR) which is a racially diverse DNA repository of over 15,000 participants. The goal of our study is: 1) to examine a racially diverse population for GR SNPs, 2) to examine the responsiveness of SNP carriers to glucocorticoids using a modified dexamethasone suppression test, and 3) to investigate the role of GR SNPs in steroid responsiveness by comparing gene expression profiles of macrophages exposed [italic]ex vivo[/italic] to glucocorticoids. Genotyping from [sim]4,000 EPR participants revealed that Black/African Americans have a lower incidence of carrying hGR9[beta]A3669G on both alleles than Caucasians (0.4% vs. 2.7%). In addition, the modified dexamethasone suppression test revealed that not only are some carriers resistant to dexamethasone, but that baseline levels of cortisol were increased in hGR9[beta]A3669G carriers as compared to non-carriers (13.5[plusmn]0.98 vs. 11.0[plusmn]0.96, p=0.06). Finally, a genome wide microarray study on macrophages isolated from three wild type participants revealed that although there is great variability in gene regulation among individuals, 447 gene probes (p[lt]0.001) were commonly regulated after dexamethasone treatment. We seek to determine how genetic variations of the glucocorticoid receptor contribute to cell type specific glucocorticoid signaling in both health and disease.[br][br]Sources of Research Support: This research was supported by the Intramural Research Program of the NIH,National Institute of Environmental Health Sciences.[br][br]Nothing to Disclose: CMJ, LM, SG, JAC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 370 311 2343 MON-454 PO02-01 Monday 2012 2012


2008 ENDO12L_MON-455 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Long-Term Cortisol Levels Measured in Scalp Hair Are Associated with Type 2 Diabetes and Cardiovascular Disease in the Elderly Laura Manenschijn, Laura A Schaap, Natasja M van Schoor, Geeske MEE Peeters, Paul Lips, Jan W Koper, Elisabeth FC van Rossum Erasmus Medical Center, Rotterdam, Netherlands; VU University Medical Center, Amsterdam, Netherlands; University of Queensland, Brisbane, Australia; VU University Medical Center, Amsterdam, Netherlands Introduction:[br]Stress is thought to be one of the main factors negatively impacting health, resulting in an increased incidence of cardiovascular disease, diabetes mellitus and obesity. The effects of stress are predominantly mediated by cortisol. Long-term pathologically elevated cortisol levels, as seen in Cushing[apos]s Syndrome, are leading to insulin resistance, dyslipidemia, increased visceral fat and hypertension, resulting in an increased cardiovascular risk. The effects of slightly elevated cortisol levels in the normal population have been previously studied by using serum and saliva cortisol measurements. These measurements are of one time point and provide no information about long-term cortisol levels. The recently developed method to measure cortisol in scalp hair provides to opportunity to measure long-term cortisol levels. We hypothesized that long-term slightly elevated hair cortisol levels are associated with cardiometabolic diseases.[br]Methods:[br]Hair samples were collected in a subgroup of 330 participants, randomly selected from a large population-based cohort study (Longitudinal Aging Study of Amsterdam). Participants with glucocorticoid use were excluded. Cortisol levels were measured in 3 cm hair segments, corresponding roughly with a period of 3 months. Cortisol was extracted with methanol and cortisol levels were measured with ELISA. Data concerning cardiovascular disease (CVD), type 2 diabetes mellitus (DM), stroke, peripheral arterial disease (PAD), cancer, osteoporosis and non-specific lung diseases were collected.[br]Results:[br]After exclusion of glucocorticoid users, hair cortisol levels were measured in 283 participants (median age 75 years, range 65-85; 66.1% women). Hair cortisol levels were significantly lower in women than in men (21.02 pg/mg hair vs. 26.29 pg/mg hair, p[lt]0.001). Hair cortisol levels were significantly higher in participants with a history of stroke (p=0.03), CVD (p=0.03), DM (p=0.02) and PAD (trend, p=0.06), but not in patients with a history of cancer, chronic lung disease or osteoporosis.[br]Conclusion:[br]Long-term cortisol levels are elevated in older adults with type 2 diabetes and cardiovascular disease, but not in those with cancer or chronic lung diseases. This suggests that elevated cortisol levels are specifically associated with cardiometabolic diseases and not with chronic diseases in general.[br][br]Nothing to Disclose: LM, LAS, NMvS, GMEEP, PL, JWK, EFCvR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1189 311 2344 MON-455 PO02-01 Monday 2013 2012


2009 ENDO12L_MON-456 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Glucocorticoid Sensitivity as a Predictor of Clinical Course in Nephrotic Syndrome Steven Ghanny, Divya Khurana, Natia Pantsulaia, Maushumi Assad, Aristotle Panayiotopoulos, Sheila Perez, Josef Michl, Duanjun Tan, Felicitas Lacbawan, Anil Mongia, Morris Schoeneman, Amrit Bhangoo, Svetlana Ten Infants[apos] and Children[apos]s Hospital at Maimonides and State University of New York (SUNY) Downstate, Brooklyn, NY; State University of New York (SUNY) Downstate, Brooklyn, NY; State University of New York (SUNY) Downstate, Brooklyn, NY; State University of New York (SUNY) Downstate, Brooklyn, NY [bold]Background:[/bold][br]Nephrotic syndrome(NS) is a heterogeneous group of disorders characterized by proteinuria, edema, hypoalbuminemia and dyslipidemia. The glomerular filtration barrier dysfunction seen in NS may be secondary to an immune disorder or due to intrinsic podocyte defects. In the case of intrinsic defects, glucocorticoid treatment is not effective, while in cases of immune mediated inflammation, there is a heterogeneous response to treatment.[br][bold]Objective[/bold]:[br]Can an in-vitro functional study to the glucocorticoid receptor correlate with clinical response to steroid treatment?[br][bold]Methods:[/bold][br]We studied 11 patients with nephrotic syndrome who were referred to pediatric endocrinology. The researchers were blinded to the clinical course of the patients. Glucocorticoid sensitivity was determined using functional study of the glucocorticoid receptor using a monocyte Fluorinated-Dexamethasone(F-Dex) binding assay to evaluate differential binding of F-Dex to the glucocorticoid receptor versus control.[br][bold]Results:[/bold][br]Out of 11 patients with nephrotic syndrome: 2 were found to be resistant, 4 had normal sensitivity and 5 had increased sensitivity to steroids. Patients who were found to be resistant to steroids were found to have minimal to no response to steroid therapy, while the patients who were found to have increased sensitivity to steroids responded very well to low doses of steroids.[br][bold]Conclusions:[/bold][br]Determination of glucocorticoid sensitivity can help predict steroid treatment response in patients with NS. This knowledge can help clinicians implement proper treatment early in the patients[apos] course and can help slow disease progression.[br][br]Nadir SJ, Saleem N, Amin F, Mahmood KT. Steroid Sensitive Nephrotic Syndrome in Paediatrics.Pak J Pharm Sci. 2011 Apr;24(2):207-10.[br][br]Nothing to Disclose: SG, DK, NP, MA, AP, SP, JM, DT, FL, AM, MS, AB, ST 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2293 311 2345 MON-456 PO02-01 Monday 2014 2012


2010 ENDO12L_MON-457 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Glucocorticoid Sensitivity as a Predictor of Asthma Management Steven Ghanny, Natia Pantsulaia, Maushumi Assad, Aristotle Panayiotopoulos, Sheila Perez, Divya Khurana, Josef Michl, Duanjun Tan, Felicitas Lacbawan, Hansoon Lee, Emily Concepcion, Amrit Bhangoo, Svetlana Ten Infants and Children[apos]s Hospital at Maimonides and State University of New York (SUNY) Downstate, Brooklyn, NY; State University of New York (SUNY) Downstate, Brooklyn, NY; State University of New York (SUNY) Downstate, Brooklyn, NY; State University of New York (SUNY) Downstate, Brooklyn, NY [bold]Background: [/bold][br]Recent data has indicated that polymorphisms in the glucocorticoid receptor gene and/or the ACTH receptor gene can influence the cause asthma. Knowledge of glucocorticoid sensitivity can be important to the management of many inflammatory diseases such as asthma, in which steroid treatment is used. The choice of corticosteroids therapy in asthmatics can be individualized as per their glucocorticoid sensitivity.[br][bold]Objective:[/bold][br]To use a monocyte Dexamethasone binding assay in asthmatic patients to predict individual glucocorticoid sensitivity.[br][bold]Methods:[/bold][br]We studied 4 patients with severe persistent asthma with multiple flare-ups on inhaled steroids +/- oral steroids. A functional study of the glucocorticoid receptor using a monocyte Fluorinated-Dexamethasone(F-Dex) binding assay was used to evaluate differential binding of F-Dex to the glucocorticoid receptor versus control. Researchers were blinded to the clinical course of the patient.[br][bold]Results:[/bold][br]One patient who was treated with inhaled steroids was found to have undetctable morning ACTH and cortisol, even after ACTH and CRH stimulation. This patient was found to have increased sensitivity to steroids on F-Dex binding assay. The other 3 patients were found to have normal morning ACTH and cortisol, but demonstrated to be resistant to steroids on F-Dex binding assay. All patients had normal glucocorticoid receptor number. This degree of resistance correlated well with asthma severity and management.[br][bold]Conclusions:[/bold][br]Determination of glucocorticoid sensitivity can help predict steroid treatment response in asthmatic patients. This knowledge can help clinicians implement proper treatment early in the patients course and can help prevent morbidity associated with poorly controlled asthma.[br][br]Tsartsali L, Papadopoulos M, Lagona E, Papadimitriou A, Kanaka-Gantenbein C, Louizou E, Kastania A, Priftis KN, Chrousos G. Association of Hypothalamic-Pituitary-Adrenal Axis-Related Polymorphisms With Stress in Asthmatic Children on Inhaled Corticosteroids.Neuroimmunomodulation. 2012;19(2):88-95.[br][br]Nothing to Disclose: SG, NP, MA, AP, SP, DK, JM, DT, FL, HL, EC, AB, ST 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2300 311 2346 MON-457 PO02-01 Monday 2015 2012


2011 ENDO12L_MON-458 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Shift Work at Young Age Is Associated with Elevated Long-Term Cortisol Levels and Body Mass Index Laura Manenschijn, Rulanda G van Kruysbergen, Frank H de Jong, Jan W Koper, Steven WJ Lamberts, Elisabeth FC van Rossum Erasmus Medical Center, Rotterdam, Netherlands; Arbo Unie Nijverdal, Nijverdal, Netherlands Introduction:[br]The incidence of obesity and other features of the metabolic syndrome is increased in shift workers. This may be due to a misalignment between the internal circadian rhythm and the behavioral rhythm. The stress hormone cortisol could play a role in this phenomenon, because it is secreted in a circadian rhythm, and long-term elevated cortisol levels lead to components of the metabolic syndrome. Our aim was to study changes in long-term cortisol levels due to shift work.[br]Methods:[br]This study consists of two parts. Part 1 (pilot study): Hair samples were collected in 33 shift workers and 89 day workers. Height and weight were measured, and BMI was calculated [1]. Part 2: Hair samples were collected in 100 males working in shifts or only during the day. Hair samples were collected together with height, weight, hip and waist circumference and questionnaires concerning diet, exercise and perceived stress. Cortisol was extracted from the hair samples with methanol and cortisol levels were measured using ELISA.[br]Results:[br]Part 1: Hair cortisol levels were higher in shift workers than in day workers: 47.32 pg/mg hair (95% CI=38.37-58.21) vs. 29.27 pg/mg hair (95% CI=26.18-33.73) (p[lt]0.001). When divided in age groups based on the median age, elevated cortisol levels were present particularly in younger shift workers: 48.53 pg/mg hair (95%CI=36.56-64.29) vs. 26.42 pg/mg hair (95% CI=22.91-30.55) in younger day workers (p[lt]0.001). BMI was increased in younger shift workers as well: 27.2 (95%CI=25.5-28.8) vs. 23.7 (95% CI=22.8-24.7) in younger day workers (p=0.001). Hair cortisol and BMI were positively correlated ([beta]=0.26; p=0.005) [1]. The results of the second part will be presented at the congress.[br]Conclusion:[br]Shift work at young adult age is associated with elevated long-term cortisol levels and increased BMI. Elevated cortisol levels and BMI may contribute to the increased cardiovascular risk found in shift workers.[br][br][1] Manenschijn L et al., J Clin Endocrinol Metab 2011; 96 (11): E1862-E1865.[br][br]Disclosures: SWJL: Scientific Board Member, Eli Lilly & Company; Speaker, Eli Lilly & Company. Nothing to Disclose: LM, RGvK, FHdJ, JWK, EFCvR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 484 311 2347 MON-458 PO02-01 Monday 2016 2012


2012 ENDO12L_MON-459 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Age-Dependent Increase in the Expression/Activity of 11[beta]-HSD1 in Muscle and Key Metabolic Tissues May Contribute to Sarcopenia Stuart Andrew Morgan, Lianne Abrahams, Gareth G Lavery, Zaki K Hassan-Smith, Paul M Stewart University of Birmingham, Birmingham, UK The pathophysiological effects of glucocorticoid (GC) excess (Cushing[apos]s syndrome) are similar to the aging phenotype. As such, we hypothesise that age-related changed in body composition (central obesity, reduced bone density, reduced muscle mass and skin thinning), and resultant chronic disease (type 2 diabetes, osteoporosis, sarcopenia and cardiovascular disease) may be caused by increased GC exposure with age. However, circulating GCs show little change with advancing age.[br]Within key metabolic tissues, murine 11[beta]-hydroxysteroid dehydrogenase type 1 (11[beta]-HSD1) converts 11-dehydrocorticosterone to the active GC, corticosterone and thus amplifies local GC action. We have previously shown 11[beta]-HSD1 expression and activity to increase with age in primary osteoblasts and skin, which may underpin age-related osteoporosis and skin thinning respectively.[br]Using C57b/6 mice (8 weeks vs. 112 weeks) we found 11[beta]-HSD1 activity to increase with age in several key metabolic tissues including: liver (2-fold, p[lt]0.05), epididymal adipose (2.7-fold, p[lt]0.05), quadriceps muscles (2.1-fold, p[lt]0.01) and soleus muscles (1.7-fold, p[lt]0.05). This was paralleled by increased mRNA expression of 11[beta]-HSD1 in epididymal adipose tissue (2.3-fold, p[lt]0.05), quadriceps muscles (6.3-fold, p[lt]0.05), tibialis anterior muscles (8-fold, p[lt]0.001) and soleus muscles (16-fold, p[lt]0.001). Furthermore, a decrease in grip-strength was observed (0.74-fold, p[lt]0.05) with age, paralleled by an age-dependent increase in the mRNA expression of the key atrophy markers MAFbx-1 (4-fold, p[lt]0.001) and MuRF-1 (1.9-fold, p[lt]0.05) in quadricep muscles. Critically, old (112-weeks) 11[beta]-HSD1 knockout mice had improved grip strength compared to old wild-type mice (1.15-fold, p[lt]0.05), suggesting GCs generated intracellularly contribute to age-related sarcopenia.[br]In summary, the expression/activity of 11[beta]-HSD1 increases with age in several key metabolic tissues, and this may accelerate sarcopenia and contribute to reduced healthy lifespan.[br][br]Nothing to Disclose: SAM, LA, GGL, ZKH-S, PMS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1435 311 2348 MON-459 PO02-01 Monday 2017 2012


2013 ENDO12L_MON-460 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Glucocorticoid and Pre-Receptor Regulation of Metabolic Function in Human Liver Maryam Nasiri, Iwona J Bujalska, Beverly A Hughes, Angela E Taylor, Paul M Stewart, Laura L Gathercole, Jeremy W Tomlinson University of Birmingham, Birmingham, UK The potent effects of glucocorticoids (GCs) on carbohydrate metabolism are well described, however, their actions upon lipid metabolism are less well understood. Patients with GC excess develop central obesity, insulin resistance and hepatic steatosis in up to 20% of cases. In rodent hepatocytes, GCs enhance insulin stimulated lipogenesis and decrease [beta]-oxidation, but studies on human primary hepatocytes have not been performed. Endogenous GCs are inactivated [italic]via [/italic]a series of enzymes including 5[alpha]-reductases (type 1 [5[alpha]R1] and 2 [5[alpha]R2]). In addition, 5[alpha]R1 and 2 generate dihydrotestosterone (DHT) from testosterone (T) and importantly both isoforms are highly expressed in human liver. We propose that GCs modulate lipid homeostasis within the human liver and that their action is regulated by 5[alpha]R activity.[br]Human hepatocytes were incubated with cortisol (0-1000nM, 24h) in the presence and absence of insulin. [italic]De novo[/italic] lipogenesis (DNL) was measured by 1-[[sup]14[/sup]C] acetate incorporation in triglyceride. The human hepatoma derived C3A cell line was used to determine the impact of 5[alpha]R over-expression on DNL. Gene expression was quantified by real-time PCR and 5[alpha]R activity by a combination of gas and liquid chromatography/mass spectroscopy.[br]In human primary hepatocytes, cortisol caused a dose dependent decrease in lipogenesis in the absence of insulin, (100% [ctrl], 80.93[plusmn]2.8% [250nM], 74.62[plusmn]4.5% [1000nM], p[lt]0.05). As expected, insulin treatment stimulated lipogenesis (100% [ctrl], 123.61[plusmn]10.1% [5nM insulin], p[lt]0.05). Interestingly, with increasing doses of cortisol, the incremental response to insulin increased (23.61[plusmn]10.7% [ctrl], 43.9[plusmn]12.7% [250nM], 66.13[plusmn]9.8% [1000nM], p[lt]0.05).[br]In C3A cells, cortisol decreased lipogenesis in a dose-dependent manner as in primary hepatocytes. 5[alpha]R2 over-expression was confirmed by real-time PCR ([gt]1000-fold) and functional activity demonstrated through increased DHT generation and cortisol clearance. In the absence of cortisol, 5[alpha]R2 transfection did not alter rates of DNL. However, in the presence of cortisol, 5[alpha]R2 completely restored rates of lipogenesis to those of untreated controls (100% [ctrl], 61.9[plusmn]7.6% [1000nM cortisol] vs. 103.8[plusmn]8.8% [5[alpha]R2+1000nM cortisol], p[lt]0.05)[br]This is the first study to define the impact of GCs upon DNL in human liver, with little evidence to suggest GC-mediated hepatic insulin resistance. Furthermore we have demonstrated the ability of 5[alpha]R to regulate the metabolic actions of GCs in the human liver.[br][br]Nothing to Disclose: MN, IJB, BAH, AET, PMS, LLG, JWT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1541 311 2349 MON-460 PO02-01 Monday 2018 2012


2014 ENDO12L_MON-461 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) [italic]Nfil3[/italic] Is a Glucocorticoid-Regulated Gene Required for Glucocorticoid-Induced Apoptosis in T-Cells Timothy James Cole, Kheng Tan, Judy Ng, Kirstyn Tamara Carey Monash University, Clayton, Australia; University of Melbourne, Parkville, Australia Glucocorticoids (GCs) have essential roles in the regulation of development, integrated metabolism, immune and neurological responses, acting primarily via the glucocorticoid receptor (GR). In most cells, GC treatment down regulates GR mRNA and protein levels via negative feedback mechanisms. In GC-treated thymocytes however, GR protein levels are maintained at a high level, increasing sensitivity of thymocytes to GCs triggering apoptosis termed Glucocorticoid-Induced Cell Death (GICD). CD4[sup]+[/sup]CD8[sup]+[/sup] double positive thymocytes and thymic NKT cells in particular are hypersensitive to GICD. Although GICD is exploited via the use of synthetic GC analogues in the treatment of hematopoietic malignancies, the downstream molecular pathway of GICD is not well understood. To explore GICD in thymocytes we have performed whole genome expression microarray analysis in mouse GR null vs wildtype thymus RNA following three hours of dexamethasone treatment. Identified and validated direct GR targets included [italic]P21[/italic] and [italic]Bim[/italic], in addition to an important transcriptional regulator [italic]Nfil3 [/italic]that has previously been associated with GICD and is essential for NK cell development [italic]in vivo[/italic]. Immunohistochemical staining of NFIL3 in whole thymus localised NFIL3 primarily to the medullary region and double immunolabelling co-localised NFIL3 to apoptotic cells and macrophages. Promoter analysis revealed a putative glucocorticoid response element [sim]5kb upstream of the [italic]Nfil3[/italic] promoter that was confirmed to bind GR by ChIP. Knock-down of [italic]Nfil3[/italic] mRNA levels to 20% of normal using specific siRNAs abrogated GICD indicating that Nfil3 is required for normal GICD in Ctll-2 T cells.[br][br]Nothing to Disclose: TJC, KT, JN, KTC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1371 311 2350 MON-461 PO02-01 Monday 2019 2012


2015 ENDO12L_MON-462 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Ligand-Induced Glucocorticoid Resistance Involves the Formation of a Novel Transcriptional Repression Complex Sivapriya Ramamoorthy, John A Cidlowski NIEHS/National Institutes of Health, Research Triangle Park, NC Glucocorticoids have been exploited in the treatment of a variety of inflammatory diseases, hematological cancers and autoimmune diseases, however subpopulations of the patients are often resistant to this steroid therapy. Unfortunately the molecular determinants of glucocorticoid resistance remain elusive. Since the level of glucocorticoid receptor (GR) is directly correlated to the magnitude of the glucocorticoid response, the level of GR is an important determinant of glucocorticoid response. One documented mechanism involved in the regulation of GR level is ligand induced down regulation of GR, but the process of homologous down regulation is still poorly understood. In the present study we have investigated the molecular mechanism responsible for the down regulation of endogenous GR mRNA. Time course studies following ligand treatment showed a rapid decrease in steady state level of GR mRNA in several human, mouse and rat cell lines as well as in various mouse tissues. A 50-70% maximal decrease in GR mRNA was observed and the down-regulation was sustained for 8-12 hours after ligand treatment. Furthermore we show that functional GR is required for this rapid repression of GR mRNA. Chromatin immunoprecipitation analysis reveals that GR binds in a ligand dependent manner to exon 6 and 8 of the GR gene and ligand induces prolonged receptor occupancy at these GR coding regions. We have also identified a functional nGRE in exon 6 of the GR gene, which might be instrumental in the active repression. Ligand induced repression of the GR gene is mediated by a NCoR1 containing repression complex at GR transcription start site. We also demonstrate that a long-range interaction occurring between this intragenic response element and the transcription start site mediated by chromatin looping is essential for constitutive expression of GR in the absence of ligand. With the addition of hormone GR is recruited to the intragenic elements leading to the disruption of the long-range chromatin loop and the exchange of an activation complex for a NCoR1 containing repression complex. This novel mechanism of repression also implies that by acquiring an intragenic regulatory element for auto-regulation, GR concentration can be coordinately down regulated with excess ligand, regardless of the various combinatorial associations of tissue specific transcription factors.[br][br]Sources of Research Support: This research was supported by the Intramural Research Program of the NIH, National Institute of Environmental Health Sciences.[br][br]Nothing to Disclose: SR, JAC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 368 311 2351 MON-462 PO02-01 Monday 2020 2012


2016 ENDO12L_MON-463 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) ACTH/cAMP-Stimulated Phosphorylation of Diaphanous 1 Regulates Cortisol Biosynthesis in H295R Human Adrenocortical Cells Donghui Li, Marion B Sewer University of California San Diego, La Jolla, CA Cortisol is synthesized by the successive action of enzymes localized in mitochondria and endoplasmic reticulum. Thus, efficient transfer of intermediates between these organelles is essential for optimal steroid production. Our lab previously reported that adrenocorticotropin (ACTH) and dibutyryl cAMP (Bt2cAMP) stimulate mitochondrial trafficking, which is integral for cortisol and dehydroepiandrosterone secretion from human adrenocortical cells. We identified RhoA and its effector diaphanous 1 (DIAPH1) as critical for regulating mitochondrial movement. To further probe the role of DIAPH1, we carried out mass spectrometric analysis which revealed that DIAPH1 is a phosphoprotein that forms a complex with several proteins, including A-kinase anchor protein 13 (AKAP13), oxysterol binding protein-related protein (ORP2), [beta]-tubulin, kinesin, and dynamin 1. By generating a phospho-specific DIAPH1 antibody against T759, we found that ACTH/cAMP signaling stimulates the phosphorylation of DIAPH1 through the activation of ERK pathway, and regulates the interaction of DIAPH1 with its binding partners. We found that the interaction of DIAPH1 with kinesin, [beta]-actin, ORP2, and AKAP13 is dependent on DIAPH1 phosphorylation. Furthermore, alanine substitution at T759 renders DIAPH1 unable to facilitate mitochondrial movement and cortisol production. Notably, the T759A mutant increased stability, suggesting a role for phosphorylation in regulating DIAPH1 degradation. Taken together, our results demonstrate that phosphorylated DIAPH1 at T759 is key for cortisol production by controlling ACTH/cAMP-stimulated mitochondrial movement and protein complex containing DIAPH1.[br][br]Sources of Research Support: This study is supported by NIH DK094151 to MBS.[br][br]Nothing to Disclose: DL, MBS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2267 311 2352 MON-463 PO02-01 Monday 2021 2012


2017 ENDO12L_MON-464 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) ORP2 Regulates Glucocorticoid Production and Cholesterol Metabolism in H295R Human Adrenocortical Cells Tamara Escajadillo, Donghui Li, Marion Sewer California State University Dominguez Hills, Dominguez Hills, CA; University of California, La Jolla, CA Steroid hormones, such as glucocorticoids, are regulators of many physiological responses. They are produced from cholesterol through a metabolic pathway that occurs in the mitochondrion and the endoplasmic reticulum, and is therefore dependent on interorganelle transfer. We have identified a macromolecular complex containing RhoA and diaphanous 1 (DIAPH1), a RhoA effector that regulates microtubule-dependent mitochondrial movement. Additional components of this complex include two members of the oxysterol-related binding protein (ORP) family, ORP2 and ORP10. Oxysterol-related binding proteins comprise a 12 member mammalian gene family of lipid transport proteins. Based on the identification of ORP2 as a DIAPH1 binding partner, we sought to determine the functional significance of ORP2 on steroidogenic gene expression and hormone production, in wild type and ORP2 knockdown H295R human adrenocortical cells. We found that ORP2 silencing resulted in an increase in the mRNA and protein expression of multiple genes, including cytochrome P450 (CYP) 11A1, CYP17A1, the melanocortin 2 receptor, and steroidogenic acute regulatory protein (StAR). In addition to genes required for cortisol biosynthesis, ORP2 suppression also led to an increase in the mRNA expression of cholesterol binding proteins Niemann Pick Type C1 and 2 (NPC1 and NPC2). Interestingly, ORP2 knockdown reduced the cAMP-stimulated secretion of cortisol, but resulted in an accumulation of cellular free cholesterol. We conclude that ORP2 modulates cortisol production by regulating the expression of genes required for cholesterol homeostasis and steroid hormone biosynthesis.[br][br]Nothing to Disclose: TE, DL, MS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2335 311 2353 MON-464 PO02-01 Monday 2022 2012


2018 ENDO12L_MON-465 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Differences in the Measurement of Cortisol in Males and Females Laura J Owen, Finlay MacKenzie, Brian G Keevil UHSM, Manchester, UK; UKNEQAS, Birmingham, UK [bold]Introduction[/bold][br]Methodological differences in the immunoassay measurement of cortisol are apparent when reviewing External Quality Assessment scheme data. A bimodal distribution is often observed, particularly for samples from female subjects. It has been postulated that some immunoassays are inefficient at removing cortisol from its binding proteins and therefore have the potential to under-recover from female samples.[br]We investigated the results obtained from two immunoassays and compared them to a liquid chromatography tandem mass spectrometry assay (LC-MS/MS).[br][bold]Methods[/bold][br]155 routine serum samples for cortisol analysis were analysed by three methods and results analysed for males and females separately. The two immunoassays utilised were the Roche E170 immunoassay and the Abbott Architect immunoassay. The LC-MS/MS assay used 20[mu]L serum to which 40[mu]L zinc sulphate was added. Following a brief vortex 100[mu]L internal standard (D4 cortisol) was added. The samples were vortexed and centrifuged then subsequently analysed on a Waters Acquity LC and Quattro Premier tandem mass spectrometer. The calibration of the LC-MS/MS assay was aligned to reference materials 192 [amp] 193.[br][bold]Results[/bold][br]The Abbott immunoassay gave a good overall comparison to LC-MS/MS; LC-MS/MS = 1.06 x Abbott [ndash] 7 nmol/L. The Roche immunoassay tended to over-estimate the cortisol; LC-MS/MS = 0.81 Roche + 19.[br]The Roche immunoassay gave a more consistent comparison to the LC-MS/MS assay between males and females than the Abbott assay. This is consistent with data from the UK NEQAS for Steroid Hormones. The comparison between the Abbott assay was different in males and females.[br][bold]Discussion[/bold][br]Users of immunoassays need to be aware of the different results that can be seen depending on the immunoassay used. Many routine assays underestimate cortisol in female subjects and this may affect result interpretation. This may be particularly important when using a generic cut-off e.g. for synacthen testing.[br][br]Nothing to Disclose: LJO, FM, BGK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 555 311 2354 MON-465 PO02-01 Monday 2023 2012


2019 ENDO12L_MON-466 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Novel Corticosteroid-Binding Globulin Variant That Lacks Cortisol-Binding Activity in a Compound Heterozygous Carrier of Other [italic]SERPINA6[/italic] Polymorphisms Lesley Ann Hill, Dimitra A Vassiliadi, Marc Simard, Aikaterini Pavlaki, Ilias Perogamvros, Dimitrios Hadjidakis, Geoffrey Lewis Hammond Child [amp] Family Research Institute, Vancouver, Canada; Attikon University Hospital, Athens University Medical School, Athens, Greece [bold]Background:[/bold] Corticosteroid-binding globulin (CBG), a serpin proteinase inhibitor family member encoded by [italic]SERPINA6[/italic], is the principal plasma binding protein for cortisol, regulating its bioavailability. Genetic CBG deficiencies that alter the cortisol-binding affinity or production of CBG are being increasingly identified. We report here on a 56-yr-old woman referred for possible secondary adrenal insufficiency, on the basis of low serum morning cortisol levels and normal plasma ACTH levels. She was suspected to suffer from a CBG deficiency, based on normal urinary and plasma free cortisol levels and a lack of consistent clinical symptoms. We therefore set out to characterize [italic]SERPINA6[/italic] polymorphisms that might contribute to a CBG deficiency in this patient.[br][bold]Methods:[/bold] Serum and DNA samples from the patient and immediate family members were obtained. The five [italic]SERPINA6[/italic] exons were PCR amplified and subjected to DNA sequencing, serum CBG level was measured by ELISA and cortisol-binding capacity assay. Recombinant CBG variants were produced to test further the effect of non-synonymous [italic]SERPINA6[/italic] single nucleotide polymorphisms on CBG production/function.[br][bold]Results:[/bold] A novel heterozygous c.1282G[gt]C transversion in exon 5 of [italic]SERPINA6[/italic], resulting in a p.Trp393Ser (W371S, CBG Athens) substitution, was identified in the proband. She was also heterozygous for two previously identified, non-synonymous SNP encoding CBG S224A and D367N (CBG Lyon) variants. The presence of both CBG D367N and W371S variants resulted in no measurable CBG cortisol-binding activity in the proband[apos]s serum despite normal CBG levels by ELISA. The proband[apos]s father and four children were heterozygous for CBG D367N, whereas the mother was heterozygous for CBG W371S [ndash] all of whom had [sim]50% lower plasma CBG cortisol-binding capacity levels. The proband[apos]s sister did not inherit the CBG D367N or W371S variants, but was heterozygous for CBG S224A and had a normal plasma CBG cortisol-binding capacity. Pedigree analysis revealed that CBG W371S segregates with Ala224, and that CBG D367N and W371S segregate separately. Recombinant CBG D367N and W371S variants exhibited no cortisol-binding activity.[br][bold]Conclusion:[/bold] We have identified a novel CBG W371S variant that lacks steroid-binding activity, which can be attributed to the fact that Trp371 directly interacts with cortisol in the binding site. The proband is the first compound heterozygous individual identified with two different cortisol-binding deficient CBG variants.[br][br]Sources of Research Support: Canadian Institutes of Health Research grant MOP-111102 awarded to GLH.[br][br]Nothing to Disclose: LAH, DAV, MS, AP, IP, DH, GLH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1850 311 2355 MON-466 PO02-01 Monday 2024 2012


2020 ENDO12L_MON-467 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) The Methyl CpG Binding Protein 2 (MeCP2) and Histone Deacetylase 1 (HDAC1) Participate in Glucocorticoid Receptor (GR)-Mediated Corticotropin-Releasing Hormone Gene (CRH) Down-Regulation Shreyas Bhave, Dharmendra Sharma, Elaine Gregg, Rosalie Uht University of North Texas Health Science Center, Fort Worth, TX; University of North Texas Health Science Center, Fort Worth, TX The hypothalamic [ndash] pituitary [ndash] adrenal (HPA) axis is a critical component of the stress response. Stress increases HPA activity, which results in increased CRH levels. It is well known that glucocorticoids down regulate crh gene expression; however the mechanism by which they do so is still poorly understood. Previous studies by Miller et al strongly suggest that HDAC1 plays a role in crh down regulation (1). In addition, McGill et al have shown that methylated CpG binding protein 2 (MeCP2) plays a role in crh repression and is present in the region of methylated CpG islands at the crh proximal promoter (2). MeCP2 is known to form a complex with HDAC1 and Sin3A. Thus, we asked whether or not GR, HDAC1, MeCP2 and Sin3A could form a complex that would participate in crh down regulation. To answer this question we used rat hypothalamic IVB cell line, used in the Miller et al report. The suitability of this line for the present study was determined, in part, by immunocytochemistry (ICC). The cells express HDAC1 and MeCP2 immunoreactivity. To determine whether GR, HDAC1 and MeCp2 could form a complex, coimmunoprecipitation was performed. Indeed the data indicated that such a complex was possible, and furthermore that GR interacts with HDAC1 and MeCp2 in the presence of Dexamethasone (Dex). To determine if the putative complex could play a role in regulating crh expression, ChIP assays were performed. Dex treatment increased occupancy of GR and MeCP2 in the region of the crh promoter. This finding prompted us to determine whether DNA methylating enzymes were also present in this region. Indeed, Dex treatment led to increased DNMT3b, but not DNMT1 and DNMT3a occupancy. The combination of increased MeCP2 and DNMT3b and the reports by McGill and Elliot strongly suggest that DNA methylation is requisite for GR-mediated crh down regulation (2) (3). Taken together; the results strongly suggest that a GR/MeCP2/HDAC1/Sin3A complex interacts with CpG islands methylated by DNMT3b to down regulate crh expression.[br][br](1) Miller Let al., Physiol Behav. 2011; 104(2):312. (2) McGill BE et al., Proc Natl Acad Sci U S A. 2006; 103(48):18267. (3) Elliott E et al., Nat Neurosci. 2010; 13(11):1351.[br][br]Sources of Research Support: NIH 1 R01 MH082900-01 awarded to RMU.[br][br]Nothing to Disclose: SB, DS, EG, RU 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 655 311 2356 MON-467 PO02-01 Monday 2025 2012


2021 ENDO12L_MON-468 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) Correlation between Nuclear Translocation of Transducer of Regulated CREB Activity (TORC) and Steroidogenic Acute Regulatory Protein (StAR) Transcription in the Adrenal of Rats Subjected to Restraint Stress or ACTH Injection Ying Liu, Victoria Poon, Ana Coello, Mark Olah, Lorna Smith, Greti Aguilera National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD Stimulation of glucocorticoid secretion during stress depends on increases in plasma ACTH leading to activation of melanocortin type 2 receptors, coupled to adenylyl cyclase/protein kinase A-dependent pathways. We have recently shown that ultradian corticosterone pulses induced by a small ACTH dose are associated with transcriptional pulses of steroidogenic proteins including StAR, P450 side chain cleavage and melanocortin receptor associated protein (MRAP). In addition to cyclic-AMP responsive element binding protein (CREB), SF1, C/EBP and GATA, the CREB coactivator, Transducer of Regulated CREB activity (TORC), has been implicated in the regulation of StAR transcription. To address the involvement of TORC in transcriptional initiation of StAR, we examined the time-relationship between nuclear translocation of TORC and induction of StAR transcription, by measuring StAR heteronuclear (hn) RNA in the adrenal zona fasciculata of rats subjected to restrain stress or ACTH injection. Restraint stress increased plasma ACTH significantly by 3 min, reaching maximal levels ([sim]400 pg/ml) at 15 min, and started to decline after 30 min. StAR hnRNA levels rose rapidly to near maximal levels at 7 min, remaining at levels about 3-fold over basal between 15 and 30 min, and started to decline at 60 min. ACTH injection (5mg, ip) increased plasma ACTH to levels in the range of those with restraint ([sim]600pg/ml at 5 min), but declined faster returning to near basal by 30 min. StAR hnRNA levels were unchanged at 5min, peaked at 15 min and declined to near basal by 60min, parallel to the decrease in plasma ACTH. For both restraint stress and ACTH injection, the increases in StAR hnRNA were preceded by decreases in cytoplasmatic and increases in nuclear dephospho-TORC 2 (3 min for restraint and 5 min for ACTH). Nuclear TORC2 levels were maximal from 7 to 30 min with stress and 5 min with ACTH injection, before declining to basal by 120 and 60 min, respectively. With restraint, nuclear phospho-CREB levels first increased at 7min, (maximum at 15 min) and slowly declined near basal levels by 120 min, while following ACTH injection levels were already maximal at 5min and had declined to basal by 60 min. The time relationship between nuclear translocation of TORC and changes in StAR hnRNA supports the involvement of the co-activator in the transcriptional initiation of StAR, and probably other steroidogenic proteins.[br][br]Nothing to Disclose: YL, VP, AC, MO, LS, GA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2046 311 2357 MON-468 PO02-01 Monday 2026 2012


2022 ENDO12L_MON-469 POSTER SESSION: Glucocorticoid Actions (1:30 PM-3:30 PM) APP695 Inactivates Steroid Synthesis in the Brain Meenakshi Bose, Manoj Prasad, Kevin J Pawlak, Jasmeet Kaur, Randy M Whittal, Himangshu S Bose Eastside High School, Gainesville, FL; Mercer University School of Medicine, Savannah, GA; University of Alberta, Edmonton, Canada Brain synthesizes not only most of the steroids, but also provides signal for different body function. Alzheimer[apos]s Disease is characterized by loss of neurons and synapses in the cerebral cortex and certain subcortical regions. Large numbers of senile plaques are characteristic of Alzheimer due to the deposition of proteins called amyloid-[beta] (A[beta]). A[beta] begins as a protein called Amyloid Precursor Protein (APP) which is expressed widely in different tissues, but is mainly concentrated in synapses of neurons. A[beta] also controls cholesterol transport and steroid synthesis in the brain. I have identified that APP695 synthesis was inhibited by puromycin and EDTA. Puromycin inhibits protein translation and EDTA arrests translocation through ER-lumen junction. Fractionation of stably expressing APP695 in COS-1 and SH-SY5Y (brain) cells, showed APP695 in ER and mitochondria, suggesting a possible relationship between APP695 and the Mitochondria-Associated-Membranes (MAM). By native gradient gel electrophoresis, I have identified a large APP695 complex. Mass Spectrometric analysis ishowed three heat shock proteins complexes-Hsp 60, Hsp 70, and Hsp 56 supporting my observations of large quantities of APP695 is associated with the mitochondria. Heat Shock Protein, Hsp 70, prevents protein folding during post-translational import into mitochondira, which is interesting in the case of APP695, because after proteolytic cleavage of APP695 by [beta]- and [gamma]- secretase forms Amyloid [beta], the protein misfolds to form a largely [beta]-pleated sheet, which is insoluble and aggregates extra cellularly. In summary, I conclude that Hsp70 helps in activation of APP695 unfolding and thus inactivates steroid synthesis in the brain.[br][br]Sources of Research Support: National Institutes of Health HD057876.[br][br]Nothing to Disclose: MB, MP, KJP, JK, RMW, HSB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 430 311 2358 MON-469 PO02-01 Monday 2027 2012


2023 ENDO12L_MON-470 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Evidence in Support of the Mismatch Hypothesis of Psychopathology in Cushing Patients after Long-Term Remission Nikolaos P Daskalakis, Nienke R Biermasz, Jitske Tiemensma, E R de Kloet, Alberto M Pereira Leiden University Medical Center, Leiden, Netherlands; Mount SInai School of Medicine, New York, NY; Leiden/Amsterdam Center for Drug Research - Leiden University Medical Center, Leiden, Netherlands Background: We have recently shown in an animal model that early-life adversity interacts with later-life stress in a manner that supports the neurodevelopmental mismatch hypothesis (1), which proposes enhanced risk for physical and mental disease in case of a mismatch between the expected, according to the degree of early-life adversity, and the actual environment.[br]Aim: to investigate the interaction of early life stress (assessed by the semi-structured interview Childhood Trauma Questionnaire) and a history of prolonged excessive exposure to cortisol in adulthood (patients in remission of the Cushing[apos]s syndrome).[br]Participants and Methods: Patients in long-term remission after treatment for Cushing[apos]s syndrome (n=60) but with persistent cognitive dysfunction and psychopathology (2,3). Healthy controls (n=37) matched for age, gender and education. Patients and controls were assessed in a sensorimotor gating test (prepulse inhibition of acoustic startle) that is interpreted as a sign of psychosis susceptibility.[br]Results: Patients treated for Cushing[apos]s syndrome had normal levels of prepulse inhibition. Early-life stress was equally prevalent in patients and controls. Early-life stress in childhood negatively affected the prepulse inhibition of controls (a significant 33% reduction, p=0.032) but did not have any effect in Cushing[apos]s patients.[br]Conclusion: Humans with early-life stress history display a better outcome in a sensorimotor gating test when exposed to cortisol excess in adulthood, in a manner that supports the mismatch hypothesis.[br][br](1)Daskalakis NP et al. Physiology [amp] Behavior (in press). (2)Tiemensma J et al., J Clin Endocrinol Metab. 2010;95:E129-41. (3)Tiemensma J et al., J Clin Endocrinol Metab. 2010;95:2699.[br][br]Nothing to Disclose: NPD, NRB, JT, ERdK, AMP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1516 312 2359 MON-470 PO03-01 Monday 2028 2012


2024 ENDO12L_MON-471 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Prevalence of Occult Cushing Syndrome in Overweight Patients Devoid of Specific Clinical Signs and Symptoms of Hypercortisolism Natalia Volkova, Maria Antonenko, Lilia Ganenko Rostov State Medical University, Rostov on Don, Russian Federation; Municipal Hospital No 4, Rostov on Don, Russian Federation Objective: the aim of the study has been to evaluate the prevalence of CS in overweight patients in the absence of specific clinical symptoms and sings of hypercortisolism.[br]Methods: 45 overweight patients were studied. Not one person had clinical features of hypercortisolism. A first screening test was the 1-mg overnight dexamethasone supression test (DST) using cutoff 50 nmol/l of cortisol supression. A second confirmatory step was performed in patients with impaired 1-mg DST and included midnight plasma cortisol concentration ([lt] 207nmol/l), plasma cortisol circadian rhythm ([lt] 50), 24-h urinary free cortisol ([lt] 180mcg/24h). Patients with impaired 1-mg DST with at least one abnormal confirmatory test result passed to the third step, the aim of which was to determine the source of hypercortisolism. It consisted of morning plasma ACTH concentration (5-46 pg/ml), 8-mg DST (autonomous secretion [gt] 27,7nmol/l) and imaging studies.[br]Results: among 43, seven patients had impaired 1-mg DST. 1 patient refused to take part in next investigations. Though 6 patients were further evaluated. 2 patients were considered as false positives of the 1-mg DST. 4 exhibited at least one additional abnormal result of the second step tests. 1 patient droped of the study. Definitive CS was diagnosed in 2 patients (4,4% of the whole series). The first one had Cushing disease (pituitary microadenoma). The second one had surgically proven adrenal adenoma. 1 patient had autonomous cortisol secretion according test results of 2, 3 steps, but nothing was found on imaging studies. The repeated tests results in 3 month didn[apos]t show autonomous cortisol secretion in this patient. The study is being continued.[br]Conclusion: preliminary results of our research show a relatively high prevalence of CS in overweight patients devoid of specific clinical signs of hypercortisolism. These findings underlines the critical necessity to finish study and get statistically proven results. Further studies are needed to compare prevalence of CS without specific clinical features between simply obese patients and obese patients with type 2 diabetes mellitus.[br][br]Nothing to Disclose: NV, MA, LG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2182 312 2360 MON-471 PO03-01 Monday 2029 2012


2025 ENDO12L_MON-472 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Development of a Clinical Prediction Score To Determine Pretest Likelihood for Endogenous Cushing Syndrome Alaa Monjed, Stan VanUum, Lisa-Ann Fraser St Joseph[apos]s Health Care, Western University, London, Canada [bold]BACKGROUND[/bold]: Cushing[apos]s syndrome (CS) is a rare disease that represents a large group of heterogeneous symptoms and signs resulting from excessive tissue exposure to high levels of glucocorticoids. Its incidence has been estimated to range from 0.7 to 2.4 per million people per year. As the clinical features of CS lack sensitivity and specificity, screening is performed in most subjects in whom the diagnosis is even remotely considered. However, none of the screening tests have ideal sensitivity or specificity. Improving clinical decision tools to determine pretest likelihood could potentially help clinicians decide when ordering screening tests would be most appropriate.[br][bold]OBJECTIVE:[/bold] To develop a clinical prediction score for estimating the pretest clinical probability of endogenous CS.[br][bold]DESIGN and METHODS[/bold]: A retrospective medical record review was conducted on all patients who underwent a 1 mg Dexamethasone suppression test between December 2003 and June 2009 in our clinic. We determined the presence or absence of clinical factors with high likelihood ratios for CS. We explored two clinical predictive scoring systems consisting of the following scores for each clinical factor (Model 1; Model 2- scores based on previously published likelihood ratios): easy bruising (1;10), facial plethora (1;3), proximal myopathy (1;8), purple striae (1;3), weight gain (1;2) and hypertension (1;4) with maximum scores of 6 and 30 points for Model 1 and 2, respectively.[br]SPSS, version 19 (Ireland) was used to perform data analysis. Two separate logistic regression models (one for each scoring model) were done with the dependent variable being final diagnosis of CS and the independent variable being the model score.Odds ratios are reported as per unit increase in model score.[br][bold]RESULTS[/bold]: We reviewed 115 charts, 19 were excluded (1 unclear diagnosis, 2 on Carbamazepine and Phenytoin, 4 CrCl[lt]60, 1 alcoholism, and 11 incomplete data). CS was diagnosed in 7 out of 96 patients (7.3%), 3 adrenal, 3 pituitary and 1 ectopic CS. Individually, none of the independent clinical variables were found to be predictive of CS. However, the OR of having CS using Model 1 was 3.37 (95%CI 1.53-7.43, p=0.003) and using Model 2 was 1.20 (95%CI 1.07-1.35, p=0.002).[br][bold]CONCLUSION[/bold]: Scoring systems that assess pretest likelihood may become helpful when deciding who to screen for CS. However, studies in larger populations are required to improve modeling, and develop clinically useful receiver operating curves.[br][br](1)Elamin MB, Murad MH, Mullan R, Erickson D, Harris K, Nadeem S, Ennis R, Erwin PJ, Montori VM. Accuracy of diagnostic tests for Cushing[apos]s syndrome: a systematic review and metaanalyses. J Clin Endocrinol Metab. 2008 May;93(5):1553-62. Epub 2008 Mar 11. (2)Valassi E, Santos A, Yaneva M, T[oacute]th M, Strasburger CJ, Chanson P, Wass JA, Chabre O, Pfeifer M, Feelders RA, Tsagarakis S, Trainer PJ, Franz H, Zopf K, Zacharieva S, Lamberts SW, Tabarin A, Webb SM; ERCUSYN Study Group. The European Registry on Cushing[apos]s syndrome: 2-year experience. Baseline demographic and clinical characteristics. Eur J Endocrinol. 2011 Sep;165(3):383-92. Epub 2011 Jun 29. (3)Nieman LK, Hall JE. Handbook of Diagnostic Endocrinology. Edition 2003. (4)Nieman LK, Biller BM, Findling JW, Newell-Price J, Savage MO, Stewart PM, Montori VM. The diagnosis of Cushing[apos]s syndrome: an Endocrine Society Clinical Practice Guideline. J Clin Endocrinol Metab. 2008 May;93(5):1526-40. Epub 2008 Mar 11.[br][br]Nothing to Disclose: AM, SV, L-AF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 524 312 2361 MON-472 PO03-01 Monday 2030 2012


2026 ENDO12L_MON-473 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Evaluation of Cushing Disease Remission Based on Serum Cortisol Dynamic Early after Transsphenoidal Surgery Fabiola Costenaro, Ticiana C Rodrigues, Guilherme Rolin, Mauro A Czepielewski Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, Brazil; Hospital de Cl[iacute]nicas de Porto Alegre, Porto Alegre, Brazil Context: Cushing[apos]s disease (CD) remission and recurrence predictors are matter of debate.[br]Objective: to evaluate the serum cortisol dynamic after transsphenoidal pituitary surgery (TSS)in predicting remission and recurrence of CD. Design and patients: a cohort of 103 CD patients from a referral center was prospectively analyzed at 111 TSS in 6.0 [plusmn] 4.8 years of follow-up.[br]Intervention: twenty patients (20 surgeries) received routine glucocorticoids in transoperative and had serum cortisol measured at 10-12 days after TSS (Protocol I). Eighty-six patients (91 surgeries)had serum cortisol measured at each 6h in the first 24h, at 48h and 10-12 days after TSS and glucocorticoid administration only if adrenal insufficiency (Protocol II).[br]Main outcomes: remission (defined as clinical hypercortisolism absence plus serum cortisol [lt] 3mcg/dl (82.8 nmol/liter) in overnight dexamethasone test and/or normal free urinary cortisol)during follow-up and recurrence (loss of remission criteria at least a year after TSS).[br]Results: we observed 80% of remission after first TSS, and 8% of the patients had recurrence. Serum cortisol nadir [lt] or equal 3.5 mcg/dl (96.6 nmol/liter) at 48h after TSS had specificity of 100% and sensitivity of 73%, and serum cortisol nadir [lt] or equal 5.7 mcg/dl (157.3 nmol/liter) at 10-12 days after surgery had specificity of 100% and sensitivity of 92% in predicting surgery remission.[br]Conclusion: at the time of the hospital discharge the cortisol nadir at 48h PO could properly predicting surgery remission. Recurrence could not be predicted. These results state the importance of serum cortisol values after TSS in predicting CD remission.[br][br]Sources of Research Support: FIPE (Fundo de Incentivo a Pesquisa do Hospital de Cl[iacute]nicas de Porto Alegre).[br][br]Nothing to Disclose: FC, TCR, GR, MAC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1215 312 2362 MON-473 PO03-01 Monday 2031 2012


2027 ENDO12L_MON-474 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Constant Light Disrupts the Circadian Rhythm of StAR and Its Regulatory Factors SF-1 and DAX1 in Rat Adrenal Glands Francesca Spiga, Shin Y Park, Jamie J Walker, Zidong Zhao, Stafford L Lightman University of Bristol, Bristol, UK The circadian rhythm of glucocorticoid hormone secretion from the adrenal cortex is regulated by the suprachiasmatic nucleus (SCN) via modulation of both hypothalamic-pituitary-adrenal (HPA) hormonal activity and direct sympathetic adrenal innervation. Disruption of normal SCN activity by exposure to constant light abolishes circadian glucocorticoid activity.[br]In the rat, the corticosterone circadian rhythm is associated with circadian expression of steroidogenic acute regulatory (StAR) protein and melanocortin type 2 receptor (MC2R) transcripts in the adrenal cortex. In this study we have investigated whether circadian expression of StAR is paralleled by circadian rhythm in SF-1 and DAX1 (StAR co-activator and co-repressor, respectively) expression and circadian expression of the MC2R accessory protein (MRAP). Furthermore, we investigated the effect of constant light on StAR, SF-1, DAX1, MC2R and MRAP.[br]Adult male rats were exposed to either a normal light-dark (LD) cycle or constant light (LL) for 5 weeks. Plasma and adrenal glands were collected either every 4 hours across the 24 hour cycle for LD rats, or at 9:00 AM and 5:00 PM for LL rats. Plasma ACTH and corticosterone levels were measured at these time points along with adrenal mRNA and protein levels of StAR, SF-1, DAX1, and MC2R and MRAP mRNA levels.[br]Circadian changes in adrenal StAR mRNA and protein levels and in MC2R mRNA in LD rats were paralleled by circadian expression of SF-1 mRNA and protein and MRAP mRNA (higher levels at the onset of the dark phase), and DAX1 mRNA and protein (higher levels at the onset of the light phase). However, exposure to constant light resulted in a disruption of circadian ACTH and corticosterone rhythms, accompanied by a loss of StAR, SF-1 and DAX1 mRNA and protein circadian rhythm and reversed MC2R and MRAP circadian rhythm.[br]Our data suggest that circadian expression of StAR is regulated by circadian expression of SF-1, DAX1 and MRAP. Furthermore, we show that a loss of the corticosterone circadian rhythm in rats exposed to constant light is associated with a disruption of the circadian rhythm of StAR, SF-1 and DAX1, and a reversed circadian rhythm of MC2R and MRAP. We propose that these changes reflect responses to the altered ACTH levels in these LL rats.[br][br]Nothing to Disclose: FS, SYP, JJW, ZZ, SLL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 239 312 2363 MON-474 PO03-01 Monday 2032 2012


2028 ENDO12L_MON-475 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Post-Surgical Recovery in Patients with Cushing Syndrome: Comparison of Patient Perceptions and Endocrinologist Perspectives Brent S Abel, Nicola N Neary, Karen Campbell, Lynnette K Nieman National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD; Research Foundation, Plymouth, MA Objective: The impaired health-related quality of life in active Cushing[apos]s syndrome (CS) improves after surgical remission, but does not normalize in all. In an open-ended survey distributed through the Cushing[apos]s Support and Research Foundation (CSRF), 84% of 94 patients described their recovery process as negative (1). The goal of this study was to better characterize the patient experience of recovery, and to compare it to physician (MD) perspectives.[br]Methods: We developed two web-based surveys: one for patients, promoted through the CSRF and the other for MDs, promoted through adult endocrinology programs in the US and UK, and the Pituitary Society. No personally-identifiable information was collected; the study was determined to be exempt from IRB review. Unpaired t-tests were used for comparisons. Median results are shown.[br]Results: 389 patients and 60 MDs completed the surveys. The self-reported etiologies of CS were pituitary (68%, n=199/291), adrenal (27%, n=80), and other (5%, n=12); remission occurred after resection of a specific lesion without bilateral adrenalectomy. MDs saw 10 patients each year with surgically-treated CS (IQR 5-20, range 0-100, n=58). MDs believed patients took 9 months (IQR 6-12, range 0-60, n=51) to discontinue cortisol replacement medication (CRM). Patients reported slightly less time (p=0.04) to discontinue CRM (8 months, IQR 5-13, range 0-42, n=142). 19% (n=5/27) of MDs described patients[apos] overall recovery experience after successful surgery as positive, 22% (n=6) negative, and 59% (n=16) mixed. Patients reported similar rates (p=0.84): 24% (n=64/264) described their experience as positive, 15% (n=39) negative and 61% (n=161) mixed. MDs thought the time to full recovery after successful surgery was 12 months (IQR 9-18, range 0-60, n=45). Patients completed the survey 36 months (IQR 17-76, range 3-405) after surgery. However, only 91 of 172 patients not on CRM (53%) reported full recovery, at 16 months after surgery (IQR 11-24, range 2-70, n=91), while 47% felt they had not fully recovered.[br]Conclusions: Patient-defined recovery takes longer than physicians realize; research is needed to identify factors that contribute to this degree of long-term impairment.[br][br](1) BS Abel et al. [ldquo]Post-surgical recovery in patients with Cushing[apos]s syndrome: Results of an open-ended survey[rdquo] P3-634, ENDO 2010.[br][br]Sources of Research Support: The intramural program of NICHD, NIH.[br][br]Nothing to Disclose: BSA, NNN, KC, LKN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1104 312 2364 MON-475 PO03-01 Monday 2033 2012


2029 ENDO12L_MON-476 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Reevaluation of the 4mg Intravenous Dexamethasone Suppression Test for Differentiation of Cushing Disease from Pseudo-Cushing Syndrome Migueline Nouvel, Muriel Rabilloud, Veronique Raverot, Chloe Pignard, Julien Vouillarmet, Charles Thivolet, Emmanuel Jouanneau, Francoise Borson-Chazot, Michel Pugeat, Gerald Raverot Groupement Hospitalier Lyon Est, Bron, France; Hospices Civils de Lyon, Lyon, France; Groupement Hospitalier Lyon Est, Bron, France; Groupement Hospitalier Lyon Sud, Pierre Benite, France; Groupemnt Hospitalier Lyon Est, Bron, France; Universit[eacute] Lyon 1, Facult[eacute] de Medecine Lyon-Est, Lyon, France [bold]Context[/bold]: Differentiating Cushing[apos]s disease (CD) from pseudo-Cushing[apos]s syndrome (PCS) is one of the most challenging problems since no biological test is yet perfect. Almost 30 years ago our group demonstrated the interest of 4mg i.v. dexamethasone suppressing test (DST) to differentiate obese patients to CD.[br][bold]Objective[/bold]: To reevaluate the diagnostic accuracy of the 4mg i.v. DST in carefully selected patients with PCS and CD[br][bold]Design[/bold]: Patients recruited from November 2008 to July 2011 were retrospectively studied.[br]The criteria for PCS were: presence of clinical and biochemical (Urinary Free Cortisol and/or 1mg DST) features compatible with CS, normal pituitary MRI and at least one year of clinical and biochemical follow-up. Diagnosis of CD has been confirmed by pathology in 29 patients operated, 3 patients with abnormal MRI were medically treated.[br]Patients underwent 4 mg dexamethasone infusion according to Abou-Samra [italic]et al[/italic] (1). The diagnosis of CD is based on absence of ACTH and cortisol suppression at 8 am on day 2.[br][bold]Results[/bold]: 68 patients (54F/14M), 32 with CD and 36 with PCS were included. Age and BMI were similar between groups but hirsutism, proximal amyotrophy, and vascular weakness were significantly more frequent in the CD group (p[lt]0.001).[br]Midnight plasma cortisol, 8 am cortisol and ACTH after 4mg DST were respectively associated with a 95.4% [86.5-99], 90.5% [81.8-96.7] and 98.4% [92.1-99.6]diagnostic accuracy. Cortisol and ACTH threshold used in our endocrine units to affirm the CD (respectively 100 nmol/L and 5 ng/l) yielded more accurate sensibility (93,8%) and specifity (86.1%) than midnight plasma cortisol cut-off of 207 nmol/l retained by the Endocrine and European Societies which was associated with a 96.8% of sensibility and 58.1% of specificity.[br][bold]Conclusion[/bold]: 4mg i.v. DST is a very easy and accurate test to distinguish CD from PCS and deserve a place in the CD diagnosis assessment.[br][br](1)Abou Samra AB, J Clin Endocrinol Metab. 1985 Jul;61(1):116-9.[br][br]Nothing to Disclose: MN, MR, VR, CP, JV, CT, EJ, FB-C, MP, GR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 436 312 2365 MON-476 PO03-01 Monday 2034 2012


2030 ENDO12L_MON-477 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) A Single Center Experience of Thromboembolism in Cushing Syndrome Selcuk Dagdelen, Safak Akin, Kadriye Aydin, Tomris Erbas Hacettepe University, School of Medicine, Ankara, Turkey Background: Hypercortisolism is a prothrombotic state. Clinical importance of the hypercoagulable state in Cushing[apos]s syndrome has recently been recognized.[br]Aim: To document the prevalance, risk factors, morbidity and mortality for thromboembolism in patients with Cushing[apos]s Syndrome (CS).[br]Materials and Methods: Consecutive patients diagnosed with endogenous CS of benign origin at our tertiary referral center (Hacettepe University Hospitals) between January 2000 and October 2010 were evaluated in a retrospective observational study. A total of 61 patients (49 female, 12 male) were included. Management of CS was conducted in compliance with standardized protocols by 5 consultants and 3 fellows of endocrinology department, without any routine anticogaulant prophylaxis.[br]Results: The mean age of the pateints were 40.8 [plusmn] 12.5. Cushing[apos]s disease (pituitary adenoma) was diagnosed in 49 patients. Nine patients were diagnosed as having adrenal adenoma, and three ectopic CS. Overall 7 thromboembolic events (prevalance: 11 %) were observed in 6 patients (5 ACTH-independent, 1 ACTH-dependent CS). All of the thromboembolic events were observed in female patients. Clinical presentation was as follows: Four events of pulmonary embolism (PE), one event of jugulary venous embolism, and two events of peripheral arterial embolism. Two of those events were complicated by in-hospital mortality. Only one of the ACTH-independent CS patients developed two episodes of thromboembolic events firstly peripheral arterial embolism which required amputation, an then PTE after adrenal surgery. One patient developed PE after transsphenoidal surgery.Two patients developed PE following the vascular catheterization.[br]Conclusion: Cushing[apos]s syndrome-associated (venous, and also arterial) thromboembolism could not be underestimated. Morbidity and mortality in our single-center 10-years experience without any prohylaxis against thromboembolism strongly suggest the necessity for thrombo-prophylaxis in CS.[br][br]Nothing to Disclose: SD, SA, KA, TE 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1842 312 2366 MON-477 PO03-01 Monday 2035 2012


2031 ENDO12L_MON-478 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Efficacy and Safety of Ketoconazole in ACTH-Dependent Cushing Syndrome Susmeeta T Sharma, Lynnette K Nieman Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD Background: Severe hepatic impairment may limit the adjunctive use of ketoconazole (KTZ) in Cushing[apos]s syndrome (CS). We evaluated its long-term efficacy and safety.[br]Methods: Retrospective chart review of ACTH-dependent CS patients (pts) on KTZ from 1982-2011. Pts were classified as ectopic ACTH syndrome (EAS) or Cushing[apos]s disease (CD) based on pathology, or as [apos]occult[apos] if testing suggested EAS but no tumor was found. Primary outcome data [urine free cortisol (UFC), serum cortisol (F), alkaline phosphatase (AP), AST, ALT] were collected before initiation, before dose change and on the final dose and analyzed as percentage of upper limit of normal (%ULN). Pre- to post-KTZ change was analyzed using Wilcoxon signed rank, McNemar[apos]s or paired Student[apos]s t-test. Differences across multiple doses were evaluated using Friedman[apos]s test followed by post-hoc test.[br]Results: Seventy-five pts (CD=36, EAS=25, Occult=14) received KTZ. Mean age at diagnosis was 38[plusmn]15 years, 52 were women. Nine pts were treated more than once (gap[ge]1 year) for N=85 KTZ treatments. Baseline (BL) median (IQR) UFC was 586.7 %ULN (299.4-1943.7), AP 67.2 %ULN (55.6-88.7), AST 58.8 %ULN (45.2-73.5) and ALT 81.8 %ULN (56.8-109.8). Initial KTZ dose was 200-1200 mg/d; maximum dose was 1800 mg/d with 0-13 dose changes per pt. UFC and F values were lower on the final dose compared to BL (P[lt]0.0001). 69.6% (39/56) pts achieved normal UFC, 27 (48.2%) on monotherapy, no difference based on BL UFC or diagnosis (EAS vs. CD) (P=NS). Systolic BP, diastolic BP and fasting plasma glucose decreased (P[lt]0.0001), fewer pts had hypertension (59.5 vs. 72.6%, P=0.0021) and diabetes (32.1 vs. 38.8%, P=0.0066) on KTZ. Potassium levels increased (P=0.049), weight remained unchanged (P=NS). AP, AST, and ALT increased significantly (P=0.0006-0.0416). Adverse events (AEs) included hepatic toxicity [n=39, 14 with severe transaminitis (AST or ALT [gt]3xULN)], gastrointestinal symptoms (GI, n=35), adrenal insufficiency, rash and fatigue. KTZ was stopped due to hepatic toxicity (n=18, 21.2%), surgical or spontaneous remission (n=40), GI (n=3), and persistent hypercortisolism (n=3). Duration on KTZ was shorter (median 14 vs. 310 days) when stopped due to AEs (P=0.0001). Final dose (median 1200, IQR=800-1200 mg/d) was similar in pts with or without hepatic toxicity (P=NS).[br]Conclusion: KTZ effectively treats hypercortisolism but leads to hepatic dysfunction in substantial number of pts; liver function tests should be monitored closely.[br][br]Sources of Research Support: This work was funded in part by the intramural program of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health.[br][br]Nothing to Disclose: STS, LKN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1319 312 2367 MON-478 PO03-01 Monday 2036 2012


2032 ENDO12L_MON-479 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Drawings Reflect a New Dimension of the Psychological Impact of Long-Term Remission of Cushing Syndrome Jitske Tiemensma, Nikolaos P Daskalakis, Else M van der Veen, Steven Ramondt, Stephanie K Richardson, Elizabeth Broadbent, Johannes A Romijn, Alberto M Pereira, Nienke R Biermasz, Adrian A Kaptein Leiden University Medical Center, Leiden, Netherlands; Leiden University Medical Center, Leiden, Netherlands; University of Auckland, Auckland, New Zealand [underline]Context and objective[/underline]: Drawings can be used to assess perceptions of patients about their disease. We aimed to explore the utility of the Drawing Test and its relation to illness perceptions, quality of life, and clinical disease severity in patients after long-term remission of Cushing[apos]s syndrome.[br][underline]Design[/underline]: Cross sectional study.[br][underline]Subjects[/underline]: We included 47 patients with long-term remission of Cushing[apos]s syndrome. Patients completed the Drawing Test, the Illness Perception Questionnaire-Revised, the Short-Form 36, the EuroQoL-5D, and the CushingQoL. The Cushing[apos]s syndrome severity index (CSI) was scored based on medical records.[br][underline]Results[/underline]: Characteristic of the drawings were strongly associated with the CSI and severity ratings of health professionals (all P[lt]0.02). In addition, patients perceived a dramatic change in body size during the active state of the disease compared to the healthy state before disease. Patients reported that their body does not completely return to the original size ([italic]i.e.[/italic] before disease) after treatment. There were no clear associations between characteristics of the drawings and quality of life (QoL) or illness perceptions. This indicates that drawings and QoL or illness perceptions do not share multiple common properties and measure different aspects/dimensions of the disease process.[br][underline]Conclusion[/underline]: Drawings reflect a new dimension of the psychological impact of long-term remission of Cushing[apos]s syndrome, because drawings do not share common properties with parameters of quality of life or illness perceptions, but do represent the clinical severity of the disease. The assessment of drawings may enable doctors to appreciate the perceptions of patients with long-term remission of Cushing[apos]s syndrome, and will lead the way in dispelling idiosyncratic beliefs.[br][br]Nothing to Disclose: JT, NPD, EMvdV, SR, SKR, EB, JAR, AMP, NRB, AAK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 291 312 2368 MON-479 PO03-01 Monday 2037 2012


2033 ENDO12L_MON-480 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Comparison of Parameters Defining Cortisol Diurnal Rhythm as Measured by Liquid Chromatography Tandem Mass Spectrometry and Immunoassay Miguel Debono, Angela Taylor, Wiebke Arlt, Richard J Ross University of Sheffield, Sheffield, UK; University of Birmingham, Birmingham, UK Background: Accurate measurements are critical for the diagnosis and treatment of endocrine disorders. Traditionally, hormones are measured by immunoassay(IA), but IA is compromised by cross reactivity and matrix effects leading to incorrect diagnosis. In contrast, Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) quantifies hormones accurately with comparable sensitivity to IA and increased specificity for steroids. We have previously reported a detailed analysis of the cortisol diurnal rhythm IA (1), and now report comparison with LC-MS/MS.[br]Methods: 20mins sampling, 24h cortisol profiles in 28 healthy volunteers were measured by: IA using a heterogeneous competitive magnetic separation assay on Bayer Immuno 1 system (intraassay CV 3[ndash]5%, interassay CV of 5[ndash]8%, and sensitivity 0.2 [mu]g/dl or 5.5 nmol/l); and LC-MS/MS using a Waters Xevo mass spectrometer with Acquity uPLC system (limit of quantitation 0.01[mu]g/dl or 0.3 nmol/l, and relative standard deviation [lt]15% at low and [lt]10% at medium and high cortisol levels).[br]Results: LC-MS/MS measured higher cortisol concentrations especially at peak cortisol levels: Passing-Bablok regression LC-MS/MS =0.2[mu]g/dl or 5.37nmol/l(95%CI 0.02 [ndash] 0.31[mu]g/dl or 0.68 [ndash] 8.52nmol/l) + 1.01(95%CI 0.99 [ndash] 1.04)(IA) and Bland-Altman analysis bias was 0.3[mu]g/dl or 7.3nmol/l(2SD -0.7 [ndash] 1.25[mu]g/dl or -20 to 34.5 nmol/l). Correlation between assays was strong(r=0.99,p[lt]0.001).[br]Key physiological cortisol parameters were defined(Geometric mean(10th [ndash] 90th percentiles)) for each assay: (LC-MS/MS vs IA)[br][bull]24-hour mean cortisol:6.4[mu]g/dl or 176nmol/l(2.8 [ndash] 13.1[mu]g/dl or 78 [ndash] 361nmol/l) vs 5.9[mu]g/dl or 164nmol/l(2.7 - 12.4[mu]g/dl or 74 [ndash] 341nmol/l)[br][bull]AUC(0-24hrs):170[mu]g/dl.h or 4697nmol/l.hr(129 - 220[mu]g/dl.h or 3560 [ndash] 6075nmol/l.h) vs 163[mu]g/dl.h or 4490nmol/l.hr(131 - 204[mu]g/dl.h or 3614 [ndash] 5632nmol/l.h)[br][bull]Peak cortisol*:21.5[mu]g/dl or 594nmol/l(15 - 35[mu]g/dl or 423 [ndash] 959) vs 18[mu]g/dl or 497nmol/l(15 - 21[mu]g/dl or 419 [ndash] 578nmol/l)[br][bull]Time of peak*:07:52(05:54 [ndash] 09:06) vs 08:01(07:02 [ndash] 09:00)[br][bull]Trough cortisol:0.8[mu]g/dl or 23nmol/l(0.4 - 1.6[mu]g/dl or 12 [ndash] 45nmol/l) vs 1[mu]g/dl or 27nmol/l(0.5 - 2.2[mu]g/dl or 13 [ndash] 61nmol/l)[br]*Average of individual peaks[br]Conclusions: This, first reported, detailed analysis of the cortisol circadian rhythm by LC-MS/MS reveals significantly higher peak cortisol concentrations at the time of maximum cortisol secretion in the early hours of the morning. These results will be important both in diagnosis and treatment of adrenal insufficiency.[br][br]Debono et al., J Clin Endocrinol Metab. 2009 May; 94(5):1548-54.[br][br]Nothing to Disclose: MD, AT, WA, RJR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1086 312 2369 MON-480 PO03-01 Monday 2038 2012


2034 ENDO12L_MON-481 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Diagnostic Utility of the Glucagon Stress Test in the Assessment of ACTH Reserve[hellip].When the Insulin Tolerance Test Is Contraindicated Jaimini Cegla, Ben Jones, Lata Seyani, Mandy Donaldson, Anthony P Goldstone, Tricia Tan Imperial College Healthcare NHS Trust, London, UK; Imperial College London, London, UK; MRC Clinical Sciences Centre, Imperial College London, London, UK Aim: The insulin tolerance test (ITT) is contraindicated in many patients with suspected ACTH deficiency and alternative tests are used to make the diagnosis. The aim of this study was to investigate the diagnostic performance of the glucagon stress test (GST) compared to the overnight metyrapone test (OMT) in patients who have contraindications to the ITT.[br]Methods: This was a retrospective dual-site study of two diagnostic tests. Data from 78 patients with suspected ACTH deficiency and contraindications to ITT were reviewed. Of these, 40 had suffered traumatic brain injury, 36 had organic pituitary disorders and 2 were classified as [apos]other[apos]. All underwent OMT and GST. Adrenal sufficiency was defined as 11-deoxycortisol greater than 200 nmol/L on OMT and cortisol greater than 440 nmol/L on GST, as per local reference ranges. Receiver-operating characteristic (ROC) analysis was performed to identify the thresholds for GST.[br]Results: There was marked discrepancy between the proportion of patients failing the OMT (39%) and GST (89%). From our data, a GST peak cortisol cut off of [ge]349 nmol/L would provide the combination of optimal sensitivity (71%) and specificity (57%), compared to sensitivity (88%) and specificity (11%) using the cortisol cut off of [ge]440 nmol/L.[br]Conclusions: We conclude that the GST overdiagnoses ACTH deficiency and is a poor test of ACTH reserve. The OMT should be used in preference to the GST to assess the HPA axis when ITT is contraindicated or resource implications limit its availability.[br][br]Nothing to Disclose: JC, BJ, LS, MD, APG, TT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1486 312 2370 MON-481 PO03-01 Monday 2039 2012


2035 ENDO12L_MON-482 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) A Revolution in the Diagnosis of Adrenal Disorders: Rapid Simultaneous Quantitation of Several Serum Steroids by Means of Tandem Mass Spectrometry (MSMS) Najiba Lahlou, Aurelie Canicio, Marc Roger, Jean Guibourdenche, Jerome Bertherat, Xavier Bertagna, Eric Clauser COCHIN Hospital, Paris, France; COCHIN Hospital, Paris, France Context: The biological diagnosis of adrenal disorders is currently based on the quantification of serum steroids using automated immunoassays (AIA). These direct assays often lack the required accuracy and precision and interlaboratory surveys show that performance characteristics of MSMS are far better than those of AIA.[br]Method: After liquid chromatography of serum extracts, steroids were quantitated on Waters QuattroPremier by either focusing several informative steroids for the diagnosis of congenital adrenal hyperplasia (CAH), or by establishing a complete steroid profile using the multisteroid-CHS-kit from PerkinElmer.[br]Subjects: Group1: 10 subjects with suspected adrenal tumour (hirsutism and abnormal ultrasound imaging); Group2: 26 neonates with suspected CAH (positive screening and/or hypoglycemia); Group3: 10 preterm neonates positive at neonatal screening; Group4: 10 female neonates with isolated clitoris enlargement.[br]Results: Group1: the complete profile including glucocorticoids, mineralocorticoids, and androgens identified in a single run the pathological secretory pattern, with increased levels of DHEA, androstenedione and cortisol precursors. Group2: 19 neonates had normal levels of cortisol and cortisol precursors, precluding the diagnosis of CAH, 6 neonates had huge excess of 17-hydroxyprogesterone, with cortisol levels, suggesting a 21-hydroxylase defect, 1 neonate had increased 11-deoxycortisol and androstenedione production, while cortisol level was low normal, data consistent with the diagnosis of 11-hydroxylase blockade. Group3: the multisteroid assay showed moderately elevated 17-hydroxyprogesterone relative to term neonates, DHEA-sulfate levels were very high, and cortisol levels were normal. In the same subjects, the steroid profile was normal at 3 months of age. Group4: the profile of 10 serum steroid was normal, precluding a disorder of sex steroid secretion.[br]Conclusion: Rapid multisteroid assays in a single run using MSMS allow confirming or rejecting the diagnosis of adrenal tumour or CAH within a few minutes. That at the same time avoids unnecessary therapy, shortens the delay of treatment starting, and cuts the investigational costs.[br][br]Nothing to Disclose: NL, AC, MR, JG, JB, XB, EC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1495 312 2371 MON-482 PO03-01 Monday 2040 2012


2036 ENDO12L_MON-483 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Clinical Application of Methods Used in the Calculation of Serum Free Cortisol (SFC) Levels in Healthy Subjects and the Critically Ill: Strengths and Limitations Laure Sayyed Kassem, Katia ElSibai, Joumana Chaiban, Baha M Arafah University Hospitals Case Medical Center, Case Western Reserve University, Cleveland, OH; Louis Stokes Cleveland Veterans Medical Center, Cleveland, OH Recent data demonstrated the serious limitations of serum total cortisol measurements in the critically ill population where hypoproteinemia is common. In such cases, measurements of SFC levels using the equilibrium dialysis (ED) method provide reliable data on adrenal function. Given the technical difficulties with the latter measurement, alternative approaches were considered and include direct ultrafiltration technique or calculating SFC levels using Coolens' formula that accounts for changes in transcortin levels. A more recent method published by Dorin incorporates changes in transcortin and albumin as well. There are no published data to validate these methods in the critically ill.[br][bold]Methods[/bold]: In the current investigation, we assess the accuracy of calculated SFC measurements using either formula in predicting ED-measured SFC levels in 122 ambulatory subjects with normal HPA function (amb-NL-HPA) and 117 critically ill patients (CI) with NL HPA. All subjects underwent low dose cortrosyn stimulation, and all available time points (n=828) were included in the analysis. Pearson correlation coefficient (r) between the calculated and measured SFC along with Bland-Altman analysis were performed.[br][bold]Results[/bold]: Both formulas provided similar results in amb-NL-HPA subjects: calculated SFC were higher (p[lt]0.01) and correlated well (r: 0.7-0.8) with measured values. Similar findings were observed in CI subjects with albumin[gt]2.5 g/dL. In contrast, both equations clearly underestimated SFC values in hypoalbuminemic CI subjects, resulting in values lower than actually measured (p[lt]0.001). The correlation coefficients between calculated and measured values in the hypoproteinemic group were lower (Coolens: r= 0.571; and Dorin: r=0.590 and data Log-transformation did not improve the correlation between the two. Bland Altman analysis of all groups done at each time point yielded very wide limits of agreement demonstrating a weak relationship between calculated and measured SFC values.[br][bold]Conclusions[/bold]: The data demonstrate that both equations provide reasonable, albeit imperfect estimates of measured SFC levels in amb-Nl-HPA subjects. However, both equations fail to predict SFC levels accurately in the clinically relevant setting of hypoalbuminemic CI subjects where total serum cortisol is not a reliable indicator of HPA function. While these equations may be useful in population-based studies, their use in individual clinical settings may not be always applicable.[br][br](1)Coolens J et al.,J Steroid Biochem 1987;26:197-202. (2)Dorin RI et al.,Clin Biochem 2009;42(1-2):64-71. (3)Hamrahian AH et al.,N Engl J Med 2004;350(16):1629-38. (4)Arafah BM,J Clin Endocrinol Metab 2006;91 (10):3725-45.[br][br]Nothing to Disclose: LSK, KE, JC, BMA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1807 312 2372 MON-483 PO03-01 Monday 2041 2012


2037 ENDO12L_MON-484 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Determining the Utility of the 60 Minute Sample in the Short Synacthen Test Ketan Dhatariya, Aditi Chitale, Patrick Musonda, Alan McGregor University of East Anglia, Norwich, UK; Norfolk and Norwich University Hospitals NHS Foundation Trust, Norwich, UK; King[apos]s College London School of Medicine, London, UK [bold]Context[/bold]: The short synacthen test (SST) was introduced over 45 years ago as a faster and safer alternative to the gold standard insulin tolerance test for the assessment of primary adrenal insufficiency. Previous work has supported the use of the 30 minute sample due to excellent correlation being documented between the 30 minute sample and the peak cortisol response to hypoglycaemia in the gold standard ITT. Despite this, and the widespread use of the SST, there remains no consensus on sampling times for the measurement of serum cortisol that best determines adrenal reserve. A Recent national survey in the UK suggest a variety of sampling times across dozens of institutions.[br][bold]Objective[/bold]: To establish whether there is any value in measuring serum cortisol at 60 minutes following administration of synacthen.[br][bold]Design: [/bold]Retrospective data analysis of 500 SST results measuring 0, 30 and 60 minute cortisol levels after administration of 250[micro]g of synacthen.[br][bold]Setting: [/bold]2 large urban National Health Teaching Hospitals in the UK[br][bold]Patients or Other Participants[/bold]: Individuals thought to have primary or secondary adrenal insufficiency.[br][bold]Intervention: [/bold]Administration of 250[micro]g of synacthen.[br][bold]Main outcome measures[/bold]: A retrospective case notes analysis was undertaken. 0, 30 and 60 minute cortisol levels were looked at to see how many people who had adrenal insufficiency at the 30 minute sample but in whom the 60 minute sample showed adequate adrenal reserve.[br][bold]Results: [/bold]The results from 324 people were analysed. The median age was 51 years (IQR 37, 65), 61% were female (IQR 56% to 66%). In 7% of people at one institution, and in 11% from the other, the 30 minute cortisol level was [apos]insufficient[apos] (i.e. [lt]550 nmol/L), but had risen to [ge]550 nmol/L at the 60 minute test. All individuals who were insufficient at 60 minutes were also insufficient at 30 minutes.[br][bold]Conclusions: [/bold]These results suggest that a significant proportion of people undergoing a SST may be inappropriately diagnosed as having adrenal insufficiency if the 60 minute sample is not measured. We suggest that the 60 minute sample is measured in all individuals having a SST to prevent unnecessary over-diagnosis of adrenal insufficiency. To our knowledge this study is the first of its kind to support the use of the 60 minute sample alongside the 0 and 30 minute samples in the conventional SST. Confirmation of our findings using a larger dataset of subjects prospectively and from multiple centres would strengthen the value of our study.[br][br]Nothing to Disclose: KD, AC, PM, AM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 11 312 2373 MON-484 PO03-01 Monday 2042 2012


2038 ENDO12L_MON-485 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Measurement of Cortisol in Scalp Hair Can Be a New Tool in the Diagnosis and Follow-Up of Patients with Addison Disease and (Cyclic) Cushing Syndrome Laura Manenschijn, Jan W Koper, Erica LT van den Akker, Loek J de Heide, Frank H de Jong, Richard A Feelders, Steven WJ Lamberts, Elisabeth FC van Rossum Erasmus Medical Center, Rotterdam, Netherlands; Erasmus Medical Center, Rotterdam, Netherlands; Medical Center Leeuwarden, Leeuwarden, Netherlands Introduction[br]The diagnosis of Cushing[apos]s Syndrome (CS) and in particular cyclic CS can be complicated. Standard screening tests for CS are the measurement of cortisol in 24-hours urine collections and in midnight saliva. In case of cyclic CS, results of these tests can be normal in between periods of hypercortisolism. The development of a method to measure cortisol in scalp hair provides the opportunity to investigate historical cortisol levels of months to years ago, with each cm of hair corresponding to a period of one month. This method has been well validated in healthy individuals and might contribute enormously in the diagnosis and follow up of patients with (cyclic) CS. Furthermore, in patients with Addison[apos]s Disease (AD), the measurement of historical cortisol levels could provide useful information concerning the disease course and effect of hydrocortisone replacement therapy. Our aim was to study whether hair cortisol levels correspond with clinical course in patients with (cyclic) CS and AD.[br]Methods[br]Hair samples were collected from 14 patients with CS, 5 cyclic CS patients and 3 AD patients. Cortisol was extracted from these samples with methanol and cortisol levels were measured using an ELISA. A group of 96 healthy non-obese individuals were used as a control group.[br]Results[br]Cortisol levels were significantly elevated in CS patients (p[lt]0.0001) and decreased in patients with AD (p=0.008) compared to healthy individuals. Hair cortisol timelines of patients with CS, cyclic CS and AD corresponded with clinical course.[br]Conclusion[br]Scalp hair can be used to evaluate the clinical course in patients with CS and AD and provides valuable information about previous cortisol exposure. This can contribute significantly in the diagnosis of cyclic CS.[br][br]Nothing to Disclose: LM, JWK, ELTvdA, LJdH, FHdJ, RAF, SWJL, EFCvR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 191 312 2374 MON-485 PO03-01 Monday 2043 2012


2039 ENDO12L_MON-486 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) The Detection of Cortisol in Human Sweat: Implications for Measurement of Cortisol in Hair Evan Russell, Gideon Koren, Michael Rieder, Stan Van Uum University of Western Ontario, London, Canada; University of Western Ontario, London, Canada BACKGROUND: Hair cortisol analysis is an effective measure of chronic stress. Cortisol is assumed to enter the hair via serum, sebum, and sweat (1), however the extent to which sweat contributes to hair cortisol content is unknown (2). Since individuals have various levels of perspiration, personal hygiene, and hair washing frequencies, it would be important to assess to what extent sweat affects hair cortisol concentrations.[br]OBJECTIVES: To determine if cortisol is present in sweat, and if its concentration would correlate with salivary cortisol concentrations. Further, to determine if exposures to a hydrocortisone solution with a sweat-like cortisol concentration would affect hair cortisol concentrations, and if washing procedures could normalize hair cortisol concentrations.[br]DESIGN and METHODS: Sweat and saliva samples were collected from 17 subjects after a period of intensive exercise, and analyzed by salivary ELISA. Subsequently, an in vitro test on hydrocortisone exposure was conducted. Residual hair samples were immersed in a 50ng/ml hydrocortisone solution for periods lasting 15 minutes to 24 hours, followed by a wash or no-wash conditions. Hair cortisol content was determined using a modified salivary ELISA protocol.[br]RESULTS: Sweat cortisol concentrations were 74.62[plusmn]41.51ng/ml (mean[plusmn]SD) and ranged from 8.16-141.7ng/ml, with the highest concentrations found in samples collected in the morning. Sweat cortisol concentrations significantly correlated with salivary cortisol concentrations (r=0.55, P [lt] 0.05) and with the log transformed time of day (r = 0.66, P [lt] 0.01). Hair exposure to a 50ng/ml hydrocortisone solution for 60 minutes or more resulted in significantly increased hair cortisol concentrations (P [lt] 0.01). Washing did not affect immersion-increased hair cortisol concentration.[br]CONCLUSION: Human sweat contains cortisol that likely contributes to hair cortisol content. Subjects with prolonged sweating at the time of hair collection may have increased hair cortisol concentrations that cannot be effectively decreased with conventional washing procedures.[br][br](1) Pragst F et al., Clin Chim Acta 2006; 370:17[mdash]49. (2) Russell E et al., Psychoneuroendocrinology 2011; doi:10.1016/j.psyneuen.2011.09.009.[br][br]Nothing to Disclose: ER, GK, MR, SVU 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1230 312 2375 MON-486 PO03-01 Monday 2044 2012


2040 ENDO12L_MON-487 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Amelioration of Non-Classical 11-Hydroxylase Deficiency with Roux-en-Y Gastric Bypass Ronald Pyram, Manish Parikh, Gul Bahtiyar, Alan Sacerdote Woodhull Medical [amp] Mental Health Center, Brooklyn, NY; State University of New York (SUNY) Downstate Medical Center, Brooklyn, NY; New York University School of Medicine, New York, NY It is well known that interventions that improve insulin sensitivity (IS), including Roux-en-Y gastric bypass (R), often ameliorate polycystic ovarian syndrome (PCOS). It is less generally appreciated that non-classic adrenal hyperplasia (NCAH) is also associated with IR and that several interventions that increase IR, including metformin, thiazolidinediones, and lifestyle alterations ameliorate the biochemical and clinical abnormalities of NCAH, just as they do in PCOS.[br]Our patient is a 39 year old deaf/mute woman with a past medical history of morbid obesity, Type 2 DM, hyperlipidemia, and male pattern alopecia with hirsutism due to non-classical 11-hydroxylase deficiency, sarcoidosis, and hyperthyroid Graves[apos] disease. On pioglitazone 45 mg/day her serum 11-deoxycortisol by liquid chromatography tandem mass spectrometry (LC-MS/MS) fell from 198 ng/dl ([lt] 51) to 26 ng/dl. Five weeks after undergoing R and off pioglitazone her serum 11-deoxycortisol was still normal at [lt]40 ng/dl. Her weight had fallen from 127.9 kg to 111.6 kg and her BMI had fallen from 44.18 kg/m2 to 39.54 kg/m2. Concurrently, her alopecia improved and her menses normalized.[br]We conclude that R may, like other interventions that improve IS, be an effective means of treating non-classic 11-hydroxylase deficiency.[br][br]Nothing to Disclose: RP, MP, GB, AS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 376 312 2376 MON-487 PO03-01 Monday 2045 2012


2041 ENDO12L_MON-488 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Ashwagandha Root in the Treatment of Non-Classical Adrenal Hyperplasia Amir Kalani, Gul Bahtiyar, Alan Sacerdote Woodhull Medical Center, Brooklyn, NY; State University of New York (SUNY) Downstate Medical Center, New York, NY; New York University School of Medicine, New York, NY; St George[apos]s University School of Medicine, St George[apos]s, Grenada [italic]Withania somnifera[/italic], commonly known as ashwagandha, is a traditional Aryuvedic medicinal plant reported to be beneficial for a variety of medical conditions owing to its antioxidant, anti-inflammatory, immune-modulating, and antistress properties. Anwer et al (2008) have also shown that [italic]Withania somnifera[/italic] (WS) increases insulin sensitivity in rats. Non-classical adrenal hyperplasia (NCAH) has been shown to be associated with insulin resistance (IR) and interventions which reduce IR, including metformin, thiazolidinediones, and weight loss ameliorate NCAH. Consequently, we hypothesized that WS could be beneficial in a patient with a non-classical adrenal hyperplasia.[br]In this case report, a 57-year-old female with non-classical congenital adrenal hyperplasia due to both 3-[beta]-ol dehydrogenase deficiency and aldosterone synthase deficiency took 400 mg WS orally twice daily for 6 months. Following this treatment there was a 31% reduction in 18-hydroxycorticosterone by mass spectrometry from 55 to 38 ng/dL (9-58), a 66% reduction in 17-hydroxypregnenolone by HPLC/mass spectrometry from 460 to 155 ng/dL (53-357), a 69% reduction in corticosterone by liquid chromatography/tandem mass spectrometry from 2416 to 748 ng/dL (130-820), and a 55% reduction in 11-deoxycortisol by liquid chromatography, tandem mass spectrometry from 89 to 40 ng/dL (12-158). The biochemical improvement was accompanied by a reduction in the amount of scalp hair loss.[br]We conclude that ashwagandha root could be used as a novel natural remedy for congenital adrenal hyperplasia if double blind, randomized control trials confirm our findings.[br][br]Anwer T, Sharma M, Pillai KK, Iqbal M. Effect of Withania somnifera on insulin sensitivity in non-insulin-dependent diabetes mellitus rats. Basic Clin Pharmacol Toxicol. 2008 Jun;102(6):498-503.[br][br]Nothing to Disclose: AK, GB, AS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 22 312 2377 MON-488 PO03-01 Monday 2046 2012


2042 ENDO12L_MON-489 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Secondary Hyperaldosteronism from Segmental Renal Artery Stenosis Responding to Partial Nephrectomy Kristin K Clemens, Andrew Fuller, Stephen E Pautler, Robert H Reid, Stan Van Uum Schulich School of Medicine and Dentistry, London, Canada; Schulich School of Medicine and Dentistry, London, Canada; Schulich School of Medicine and Dentistry, London, Canada Introduction[br]Hypertensive hypokalemia may arise from primary hyperaldosteronism or from secondary hyperaldosteronism in the setting of an activated renin-angiotensin system. We present a patient with hypertension, hypokalemia and secondary hyperaldosteronism from segmental renal artery stenosis.[br]Clinical Case[br]A 50 year old lady was referred for fatigue, parasthesia, weakness, and progressive hypertension and hypokalemia requiring previous hospitilization. Past trials of amlodipine and bisoprolol had not controlled her blood pressure, and diuretic therapy exacerbated her hypokalemia. On atacand 32 mg daily, her blood pressure was between 150-190/80 mm Hg and she required 20-40 meq of daily potassium supplementation to keep her serum level above 3.5 mmol/L.[br]On our assessment her blood pressure was 153/86 mm Hg with a heart rate of 66 beats per minute. BMI was 30.9 kg/m[sup]2[/sup]. She had an otherwise normal exam with no evidence of hypertensive retinopathy, and no bruits over her carotid, renal or femoral arteries.[br]Her bloodwork on atacand revealed a non-suppressed aldosterone of 285 pmol/L (96-522 pmol/L) with a renin of 5.2 ng/L (1.1[ndash]20.2 ng/L). Aldosterone secretion was in the normal range on 24 hour urine collections.[br]A lack of blood pressure response to the addition of another antihypertensive medication together with increasing potassium requirements raised our clinical suspicion of renovascular disease. A CT renal angiogram demonstrated a diminutive, atretic branch of the left main renal artery supplying an atrophic left lower pole with a solitary collecting system. She had impaired left kidney function at 31.5% of predicted on renogram. She was started on eplerenone therapy and titrated to a dose of 50 mg daily.[br]The patient underwent a laparoscopic partial nephrectomy of the atrophic left lower pole without complication. Final pathology was consistent with atrophic tubules and interstitial fibrosis. Post operatively, her potassium normalized and her supplementation was discontinued. On low dose atacand her blood pressure was 130/80.[br]Conclusion[br]When patients present with progressive hypokalemic hypertension, clinical suspicion of renal artery stenosis should remain high. Surgical intervention can be curative for many patients.[br][br]Nothing to Disclose: KKC, AF, SEP, RHR, SVU 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 652 312 2378 MON-489 PO03-01 Monday 2047 2012


2043 ENDO12L_MON-490 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Bilateral Cervical and a Functional, Occult, Intrapericardial Paraganglioma (PGL) in a Patient with SDHC Mutation Bindiya T Magoon, Arthur Tischler, Hassan Rastegar, Richard Wein, Carl Heilman, James Udelson, Michael England, Ronald M Lechan Tufts Medical Center, Boston, MA; Tufts Medical Center, Boston, MA; Tufts Medical Center, Boston, MA; Tufts Medical Center, Boston, MA; Tufts Medical Center, Boston, MA; Tufts Medical Center, Boston, MA; Tufts Medical Center, Boston, MA Background: PGL3 or SDHC mutations present almost exclusively as head and neck PGLs. Only rare cases have been described in literature where they have been associated with adrenal pheochromocytomas or abdominal PGLs. We describe a patient with a SDHC mutation with bilateral cervical paragangliomas and a functional, intrapericardial paraganglioma.[br]Case: A 32 yr. old male with difficult to control hypertension on multiple antihypertensive medications reported episodic headaches, sweating and palpitations for several years. On presentation, he had hoarseness of voice, slurred speech, impaired tongue movement and difficulty swallowing with 115 lb weight loss over a period of 6 months. On physical examination, he had a moderately elevated BP with multiple left cranial nerve palsies. MRI of his neck showed a right carotid body tumor and a left glomus jugulare tumor. Biochemical profile revealed elevated 24 h urinary dopamine (2204 mcg, nl 52-480), norepinephrine (491 mcg, nl 15-100), and normetanephrine (2725 mcg, nl 35-482). PanCT and MIBG scan were negative for other paragangliomas. Octreoscan showed uptake on both sides of the neck in the region of the known PGLs and in the lower retrosternum, suggestive of a lesion overlying the heart. Cardiac MRI showed a 6 cm mass in the atrioventricular groove. The patient underwent uneventful surgery with dexmedetomidine as an anesthetic coadjuvant. A highly vascularized, 6 cm tumor adherent to the right atrium, ventricle and aortic wall was excised. Pathology revealed a tyrosine hydroxylase and chromogranin A-positive paraganglioma with neuromelanin pigmentation that was SDHB immunostaining negative. Genetic screening revealed a c.397 C[gt]T, p133 R[gt]X nonsense mutation on chromosome 1. Catacholamine levels normalized following surgery.[br]Conclusions: Mediastinal and cardiac PGLs are relatively rare tumors with less than 100 cases described in literature. With the availability of genetic testing for SDH mutations, a strong association between cardiac paragangliomas and SDHB and SDHD mutations has been recognized. This case report indicates that patients with SDHC mutations may also develop cardiac paragangliomas, confirming the recent single case report by Vandy et al (1). Since head and neck PGLs rarely secrete catacholamine, patients with SDHC mutations and evidence for catecholamines excess should undergo radiologic screening for paragangliomas in the chest and abdomen.[br][br]1)Vandy FC et al. Synchronous carotid body and thoracic paraganglioma associated with germline SDHC mutation. J of Vasc Surg 2011 53(3)805-7.[br][br]Nothing to Disclose: BTM, AT, HR, RW, CH, JU, ME, RML 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1017 312 2379 MON-490 PO03-01 Monday 2048 2012


2044 ENDO12L_MON-491 POSTER SESSION: Adrenal Disorders (1:30 PM-3:30 PM) Risk of Iatrogenic Cushing Syndrome and Adrenal Suppression in Antiretroviral Regimes Including CYP450 3A4 Inhibitors Frances McManus, Beth White, Alisdair MacConnachie, Colin Perry Western Infirmary, Glasgow, UK; Gartnavel General Hospital, Glasgow, UK A 60 year old female from Zimbabwe with an eight year history of HIV infection presented with malaise and facial swelling. She had been established on antiretroviral treatment (Kivexa (lamivudine/abacavir) and Kaletra (lopinivir/ritonavir)) since shortly after her initial diagnosis.[br]One month earlier she had received an injection of intramuscular triamcinolone acetate to treat an inflammatory polyarthropathy. She was also prescribed inhaled salmeterol xinafoate 50mcg/fluticasone propionate 500mcg per dose, one puff twice daily for asthma.[br]On examination, she displayed classical features of glucocorticoid excess with striae, proximal myopathy and centripetal obesity. She was hypertensive, but not hyperglycaemic.[br]24 hour urine collection revealed no detectable cortisol. A short synacthen test revealed an undetectable baseline cortisol rising to 53mmol/L at 30 minutes.[br]The working diagnosis was iatrogenic Cushing[apos]s syndrome, with concurrent adrenal suppression secondary to exogenous steroids in combination with the protease inhibitor (PI) ritonavir, a potent CYP 3A4 pathway inhibitor. Ritonavir was discontinued and due to her adrenal suppression, low dose steroid replacement was initiated (prednisolone 2.5mg daily).[br]While intramuscular and inhaled or intranasal steroid use rarely lead to iatrogenic Cushing[apos]s syndrome in the normal population, this case illustrates the clinical significance that inhibition of the CYP 3A4 pathway has on the metabolism of exogenous steroid preparations. Ritonavir is a PI used in the treatment of HIV infection to [ldquo]boost[rdquo] other PIs via inhibition of cytochrome P450 3A4 activity. Boosted PIs are used commonly as part of antiretroviral regimens. In Glasgow, 33% of patients treated for HIV infection are on a regimen which includes them. Caution should be exercised before prescribing steroid treatment in these patients, even via routes thought to have less risk of causing adrenal suppression, and consideration given to adrenal suppression during intercurrent illness.[br][br]Nothing to Disclose: FM, BW, AM, CP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 855 312 2380 MON-491 PO03-01 Monday 2049 2012


2045 ENDO12L_MON-492 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) The Ins and Outs of OTX2: Assessment of Intrinsic and Extrinsic Functions in Pituitary Organogenesis Amanda H Mortensen, Thomas Lamonerie, Sally A Camper University of Michigan, Ann Arbor, MI; University of Nice, Nice, France Patients heterozygous for mutations in the [italic]OTX2[/italic] transcription factor gene have variable features that can include cranio-facial and ocular malformations and pituitary-specific defects. Those with pituitary anomalies have a range of endocrinological manifestations that include ectopic pituitary, hypopituitarism, isolated growth hormone deficiency, and multiple hormone deficiencies. Phenotypic variability also occurs among patients with the same mutation, implying that variation in other genes influences the clinical presentation (Reviewed in 1). Mice heterozygous for [italic]Otx2[/italic] loss of function also exhibit a range of phenotypes that are influenced by genomic variation (2,3). To understand the mechanism whereby OTX2 deficiency influences pituitary development we established the expression pattern of OTX2 in mouse pituitary ontogeny (4) and generated tissue-specific knockouts. At e10.5 OTX2 is expressed in both the ventral diencephalon, which gives rise to the posterior lobe (neurohypophysis), and Rathke[apos]s pouch, the primordium for the adenohypophysis (anterior lobe) and intermediate lobe. Subsequently, OTX2 expression is restricted to the posterior lobe and intermediate lobe. No OTX2 protein is detected by immunohistochemistry in newborn mouse pituitary glands. We hypothesize that OTX2 deficiency causes anterior pituitary hypoplasia by affecting inductive signaling in the ventral diencephalon. To test this idea we generated a neural ectoderm-specific knockout by crossing [italic]Otx2[/italic][sup]flox/flox[/sup] transgenic mice (5) with [italic]Nkx2.1-Cre[/italic] transgenic mice (6). The [italic]Otx2[/italic][sup]flox/flox[/sup], [italic]Nkx2.1-Cre[/italic] mice have poor invagination of the ventral diencephalon at e12.5, and Rathke[apos]s pouch is smaller. These preliminary results support the idea that [italic]Otx2[/italic] expression in the neural-ectoderm is important for induction of Rathke[apos]s pouch [ndash] an extrinsic effect. OTX2 is involved in WNT, FGF and BMP signaling in other tissues, and all of these pathways are required in the neural ectoderm for normal growth of Rathke[apos]s pouch (7,8). To investigate whether OTX2 also has an intrinsic role in Rathke[apos]s pouch, we crossed the [italic]Otx2[/italic]flox/flox mice with [italic]Pitx2-cre[/italic] transgenic mice (9). These complimentary tissue-specific knockouts will decipher the intrinsic and extrinsic roles of [italic]Otx2[/italic] on Rathke[apos]s pouch growth and hormone producing-cell differentiation. This work will establish the molecular mechanism of OTX2 action in pituitary gland development and the etiology of hypopituitarism in human patients.[br][br](1) McCabe MJ et al., Best Pract Res Clin Endocrinol Metab. 2011; 25:115-24. (2) Matsuo I et al. Genes Dev. 1995 9:2646-58. (3) Hide T et al., Development. 2002;129:4347-57. (4) Mortensen AH et al, Physiol Genomics. 2011; 43:1105-16. (5) Fossat N et al., EMBO Rep. 2006; 7:824-30. (6) Xu Q et al. J Comp Neurol. 2008; 506:16-29. (7) Brinkmeier et al., Dev Biol. 2007; 311:396-407. (8) Davis SW, Camper SA. Dev Biol. 2007; 305:145-60. (9) Liu C, et al., Development. 2002;129:5081-91.[br][br]Sources of Research Support: NIH R37HD30428, R01HD34283.[br][br]Nothing to Disclose: AHM, TL, SAC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1014 313 2381 MON-492 PO20-01 Monday 2050 2012


2046 ENDO12L_MON-493 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) LHX4 Dysfunction Causes Pituitary Hypoplasia by Permitting Ectopic Expression of the Cell Cycle Inhibitor p21 Peter Gergics, Michelle L Brinkmeier, Sally A Camper University of Michigan, Ann Arbor, MI Pituitary gland development requires synchronous regulation and function of homeodomain transcription factors including LHX4, LHX3, OTX2, PITX2, PROP1 and POU1F1 (1). Defects in these genes cause profound pituitary hypoplasia. Little is known about the mechanisms underlying pituitary hypoplasia, although there are examples of reduced proliferation and increased apoptosis (2, 3, 4, 5). Abnormalities in cell cycle regulators such as cyclins, cyclin dependent kinase inhibitors, and the retinoblastoma tumor suppressor gene (Rb) can cause pituitary hypoplasia or hyperplasia (6), but the links between these key cell cycle regulators and the transcription factors that affect cell proliferation and growth are unknown. To clarify the mechanism of cell cycle misregulation underlying pituitary hypoplasia we examined expression of a variety of cell cycle regulators in [italic]Lhx4[/italic], [italic]Lhx3[/italic], and [italic]Pitx2[/italic] mutant mice. Each of these genetic defects causes profound hypoplasia of the pituitary primordium, Rathke[apos]s pouch. [italic]Lhx4[/italic] mutants exhibit a striking expansion in expression of p21 (CDKN1A), a cyclin-dependent kinase inhibitor that normally regulates cell cycle progression and cellular senescence. Normally p21 expression is restricted to the sites of pouch closure, in a region of cell death, while broad, ectopic expression is detected in [italic]Lhx4[/italic] mutants. The domain of ectopic p21 expression is also marked by phosphorylated-[gamma]-histone 2A.X (P-[gamma]H2A.X), a senescence marker. Cyclin D1 (CCND1) normally drives the G[sub]1[/sub]-S transition and is expressed in a mutually exclusive pattern with p21. Cyclin D1 expression is reduced in [italic]Lhx4[/italic] mutants. Alterations in CCND2/E, pRB, Ki67, and p57 expression in [italic]Lhx4[/italic] mutants were less striking than p21, P-[gamma]H2A.X, and cyclin D1. Functional studies in the pituitary cell line [alpha]T3-1 show that LHX4 suppresses p21 transcription via evolutionarily conserved sequences near the p21 promoter. Our studies support the idea that [italic]Lhx4[/italic] regulates the cell cycle in early pituitary formation by restricting p21 expression. Failure to do this leads to the reduced proliferation, senescence and apoptosis, explaining the pituitary hypoplasia and reduced function. We observed reduced Cyclin D1 expression in [italic]Lhx3[/italic] and [italic]Pitx2[/italic] mutants, suggesting common mechanisms of hypoplasia. These studies are yielding a better understanding of how expansion of early pituitary progenitor cells is controlled and how these mechanisms relate to regulation of differentiated cell growth (7).[br][br]1. Kelberman D et al., Endocr Rev, 2009; 7:790-829. 2. Raetzman LT et al., Development, 2002; 18:4229-4239. 3. Ellsworth BS et al., Dev Biol, 2008; 1:118-129. 4. Charles MA, Suh H et al., Mol Endocrinol. 2005;19:1893-903. 5. Ward RD et al, Mol Endocrinol. 2006;20:1378-90. 6. Quereda V, Malumbres M. J Mol Endocrinol 2009; 42:75-86. 7. Bilodeau S et al., Mol Cell Biol, 2009; 7:1895-1908.[br][br]Sources of Research Support: NIH R01HD34283 and R37HD30438.[br][br]Nothing to Disclose: PG, MLB, SAC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1255 313 2382 MON-493 PO20-01 Monday 2051 2012


2047 ENDO12L_MON-494 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) [italic]OTX2[/italic] Gene Mutations Are Rare in Patients with Combined Pituitary Hormone Deficiency and/or Midline Cerebral Defects Nathalia Novaretti, Sabrina S Paulo, Antonio C Santos, Ayrton C Moreira, Carlos E Martinelli, Jr, Maki Igarashi, Tsutomu Ogata, Maki Fukami, Margaret Castro, Sonir R Antonini School of Medicine of Rebeir[atilde]o Preto - University of S[atilde]o Paulo, Rebeir[atilde]o Preto, Brazil; School of Medicine of Rebeir[atilde]o Preto - University of S[atilde]o Paulo, Rebeir[atilde]o Preto, Brazil; National Research Institute for Child Health and Development, Tokyo, Japan; Hamamatsu University School of Medicine, Hamamatsu, Japan [bold]Background: [/bold]OTX2 controls variable development processes, including anterior brain, eye and pituitary formation. [italic]OTX2[/italic] mutations have been frequently identified in patients presenting with anophthalmia, microphthalmia and coloboma spectrum. On the other hand, [italic]OTX2[/italic] mutation was identified in a single patient with combined pituitary hormone deficiency (CPHD) without midline cerebral defects (MCD) or eye abnormality.[br][bold]Objective: [/bold]To investigate the presence of [italic]OTX2[/italic] mutations in patients with CPHD presenting with different degrees of MCD.[br][bold]Patients and Methods: [/bold]We evaluated 117 Patients with CPHD and/or MCD from a single University center, including 61 with CPHD without MCD, 32 with septo-optic dysplasia (SOD), 15 with holoprosencephaly (HPE) and 9 with isolated anomaly of corpus callosum (ACC). All patients underwent clinical, laboratory and CNS MRI evaluation. [italic]OTX2[/italic] coding and boundary regions were sequenced. [italic]In silico[/italic] analysis: Polyphen and Fruitfly Splice Site Prediction. Transactivating activity of a mutant OTX2 was examined [italic]in vitro[/italic] by luciferase assays using reporter vectors containing the promoter sequences of IRBP, HESX1, POU1F and GNRH1.[br][bold]Results: [/bold]Mean age (years) at diagnosis was 5.8 (0.7 [ndash] 14.4) in SOD patients, 9 (6.5 [ndash] 11.7) in patients ACC, and 10.7 (1.2 [ndash] 27) in CPHD without MCD patients. HPE was antenatally diagnosed in most of patients. Endocrine deficiencies (CPHD with or without ADH deficiency or isolated GH deficiency-GHD) were found in 69%, 67%, and 53% in SOD, ACC, and HPE patients, respectively. A new[italic] OTX2 [/italic]variant c.757G[gt]A (p.A253T) was identified in a CPHD patient presenting with mild ocular hypotelorism and normal visual function, hypoplastic anterior pituitary and ectopic posterior pituitary. [italic]In vitro[/italic] analysis showed no differences between the wild-type OTX2 and Ala253Thr OTX2. The new [italic]OTX2[/italic] variant c.444G[gt]C (p.P148) was identified in a SOD patient and the intronic variant c.*19T[gt]A was identified in a SOD patient and in a CPHD patient. None of these 3 new [italic]OTX2[/italic] gene variants were found in 120 Brazilian healthy controls neither they have been described in the 1000genomes project. [italic]In silico[/italic] analysis showed that none of the new [italic]OTX2[/italic] variants are likely to impair splicing.[br][bold]Conclusions: [/bold]Three new [italic]OTX2[/italic] variants were identified only in patients with CPHD and/or MCD. [italic]In vitro[/italic] and [italic]in silico[/italic] analysis revealed that these variants are likely to be non functional variants. Therefore, mutations in [italic]OTX2[/italic] gene appear to be a rare cause of CPHD and/or MCD.[br][br]Sources of Research Support: FAPESP - Brazil. CNPq - Brazil.[br][br]Nothing to Disclose: NN, SSP, ACS, ACM, CEM, MI, TO, MF, MC, SRA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1947 313 2383 MON-494 PO20-01 Monday 2052 2012


2048 ENDO12L_MON-495 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Genetic Screening in a Cohort of 2030 Patients with Congenital Hypopituitarism; Current Knowledge and Future Directions Kyriaki S Alatzoglou, James PG Turton, Daniel Kelberman, Marc J McCabe, Louise C Gregory, Emma A Webb, David EG McNay, Kathryn S Woods, Ameeta Mehta, Mehul T Dattani UCL Institute of Child Health, London, UK; National Institute for Medical Research, London, UK; UCL Institute of Child Health, London, UK [bold]Background[/bold]: Congenital hypopituitarism (CH) encompasses a spectrum of phenotypes ranging from isolated to multiple pituitary hormone deficiencies with/without associated midline or ocular defects and/or structural pituitary abnormalities. Known genetic factors have so far explained a relatively small percentage of cases with varying results depending on the cohort screened. We analysed the results of genetic screening in a large cohort of patients with CH performed in our centre.[br][bold]Patients and Methods: [/bold]Over 15 years we screened samples from 2366 individuals, referred from national and international centres, including 2032 patients with CH and 334 unaffected family members. Our cohort included 540 patients with combined pituitary hormone deficiencies (CPHD), 522 with variable hypopituitarism, 540 with septo-optic dysplasia (SOD) and its variants (26%), 368 with isolated GH deficiency (IGHD) (18%) and 62 with midline clefts including holoprosencephaly (3%) In 17.4% (n=362) there was a positive family history. According to phenotype, patients were screened for mutations in known genetic factors including [italic]HESX1, SOX2, SOX3, OTX2, LHX3/LHX4, SIX3, SHH, GLI2, CHD7, FGF8/FGFR1, KAL-1, PROK2/PROKR2, PROP1, POU1F1, GH1, GHRHR.[/italic][br][bold]Results: [/bold]In patients with SOD [italic]HESX1[/italic] mutations were identified in 1.3% (7/540), whilst changes in [italic]SOX3[/italic] dosage were rare, identified in 6 of the probands. Mutations in [italic]SOX2[/italic] (n=13) and [italic]OTX2[/italic] (n=4, including deletions) were also rare in patients with SOD, but accounted for almost 28% and 4.5% respectively of patients with severe eye phenotypes (13/45 and 4/45). Variations in [italic]FGF8 [/italic]were identified in less than 1% whilst [italic]PROKR2[/italic] variations were the commonest (2% of screened patients), but their contribution to phenotype remains to be established. In patients with CPHD, [italic]PROP1[/italic] mutations were identified in 18 patients (3.3%) and [italic]POU1F1[/italic] mutations in 12 (2.2%), with higher percentages in familial cases. Genetic changes ([italic]GH-1[/italic], [italic]GHRHR[/italic]) were identified in 34 patients (9%) with IGHD.[br][bold]Conclusion:[/bold] With the current methods of screening, known genetic factors account for 5% of cases of CH (n=106); this percentage increases in familial cases and if specific phenotypes are taken into account. Other yet unidentified factors are therefore likely to be implicated in the etiology of CH, either genetic or environmental, or a combination of both. Next generation sequencing in combination with careful phenotyping will be critical in the identification of further factors.[br][br]Nothing to Disclose: KSA, JPGT, DK, MJM, LCG, EAW, DEGM, KSW, AM, MTD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1908 313 2384 MON-495 PO20-01 Monday 2053 2012


2049 ENDO12L_MON-496 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Variations in [italic]PROKR2[/italic], but Not [italic]PROK2[/italic], Are Associated with Hypopituitarism and Septo-Optic Dysplasia Mark James McCabe, Carles Gaston-Massuet, Louise Cheryl Gregory, Vaitsa Tziaferi, Kyriaki S Alatzoglou, Oualid Sbai, Philippe Rondard, Koh-Hei Masumoto, Mamoru Nagano, Yasufumi Shigeyoshi, Marija Pfeifer, Tony Hulse, Charles Buchanan, Nelly Pitteloud, Juan-Pedro Martinez-Barbera, Mehul T Dattani UCL-Institute of Child Health, London, UK; UCL-Institute of Child Health, London, UK; Universite Montpellier, Montpellier, France; School of Medicine, Kinki University, Osaka, Japan; University Medical Centre, Ljubljana, Slovenia; Guy[apos]s and St Thomas[apos] Hospital, London, UK; King[apos]s College Hospital, London, UK; University of Lausanne, Lausanne, Switzerland [bold]Background and Aims: [/bold]Loss-of-function mutations in [italic]PROK2 [/italic]and [italic]PROKR2 [/italic]have been implicated in Kallmann syndrome (KS), characterized by hypogonadotropic hypogonadism and anosmia. Recent evidence suggests overlapping phenotypes/genotypes between KS and congenital hypopituitarism (CH) including septo-optic dysplasia (SOD). We therefore aimed to screen a cohort of patients with complex forms of CH with and without SOD for mutations in [italic]PROK2 [/italic]and [italic]PROKR2 [/italic]and study in detail the hypothalamic-pituitary structure and function in the [italic]Prokr2[sup]-/-[/sup][/italic] mouse model.[br][bold]Methods:[/bold] DNA from 421 patients with CH/SOD was amplified by PCR and directly sequenced for [italic]PROK2[/italic] and [italic]PROKR2[/italic]; the largest such screen to date. Functional testing of novel variants was conducted [italic]in vitro[/italic] by intracellular Ca[sup]2+[/sup] mobilization and phospho-inositol turnover assays. [italic]In situ[/italic] hybridization was performed using probes for various hormone markers to analyse hypothalamo-pituitary development in the [italic]Prokr2[sup]-/-[/sup][/italic] mice.[br][bold]Results:[/bold] Five [italic]PROKR2 [/italic](no [italic]PROK2[/italic]) variants were detected predominantly in heterozygosity in ten patients with SOD/CH, including hormone deficiencies ranging from isolated GHD to panhypopituitarism and diabetes insipidus. These included the novel p.G371R (n=1) and the previously reported p.A51T (n=1), p.R85L (n=1), p.L173R (n=4), and p.R268C (n=3). These patients were clear of variations in any known KS or CH/SOD genes. Surprisingly, one patient with SOD was heterozygous for the functionally significant p.L173R variant whereas the phenotypically unaffected mother was homozygous.[br]The p.G371R change led to no loss of [italic]in vitro[/italic] function whereas p.R85L, p.L173R and p.R268C are known functionally deleterious variants. Pituitary and hypothalamic development and terminal cell differentiation were predominantly normal apart from a reduction in LH in the [italic]Prokr2[sup]-/- [/sup][/italic]mice.[br][bold]Conclusions:[/bold] Our data show that loss of function variants in heterozygosity or homozygosity can be associated with KS (shown previously), CH and SOD; there are no obvious genotype-phenotype correlations. Thus the role of PROKR2 in the etiology of these disorders is unclear and the variants identified herein are unlikely to be implicated in isolation in these disorders. Other genetic or environmental modifiers may also impact on the etiology.[br]Patients with sequence variations in [italic]PROKR2 [/italic]therefore represent a unique cohort worthy of careful further investigation that may uncover a possible oligogenic basis not only to KS, but also to CH and SOD.[br][br]Nothing to Disclose: MJM, CG-M, LCG, VT, KSA, OS, PR, K-HM, MN, YS, MP, TH, CB, NP, J-PM-B , MTD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1472 313 2385 MON-496 PO20-01 Monday 2054 2012


2050 ENDO12L_MON-497 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Pathophysiological Mechanism of Dominant Pit-1 Mutations: (R271W and K216E) Sumito Dateki, Fredric E Wondisford, Sally Radovick Johns Hopkins University School of Medicine, Baltimore, MD; Johns Hopkins University School of Medicine, Baltimore, MD [bold]Context: [/bold]Pituitary-specific transcription factor-1 (Pit-1) is a POU-homeodomain transcription factor that induces differentiation of GH-, PRL-, and TSH-producing cells in the anterior pituitary. Pit-1 usually binds to multiple sites on target genes, and dimerization of Pit-1 on DNA elements is thought to be important for high-affinity DNA binding. Although most Pit-1 mutations are autosomal recessive and affect normal DNA binding, dominant negative Pit-1 mutations that bind DNA normally also have been reported.[br][bold]Objective:[/bold] To understand the nature of Pit-1 complex on DNA to gain insight into pathophysiological mechanisms of two dominant Pit-1 mutations (R271W and K216E) found in humans with combined pituitary hormone deficiencies (CPHD).[br][bold]Methods:[/bold] Electrophoretic mobility shift assays (EMSA) were performed to investigate the DNA binding properties of these two mutant proteins on multiple Pit-1 binding sites in the GH and PRL promoters. The target sequences are as follows; GH-1 (5[apos]-CTATACATTTATTCATGG-3[apos]), GH-2 (5[apos]-CTTCTAAATTATCCATCA -3[apos]), PRL-1P (5[apos]-GATTATATATATTCATGA -3[apos]), PRL-1D (5[apos]-GCATTAAAAAATGCATTT -3[apos]). Proteins for EMSAs were obtained by T7 TNT-coupled reticulocyte lysate and His-tagged protein expression and purification systems.[br][bold]Results: [/bold]Wild-type Pit-1 protein bound to GH-1 and PRL-1P elements as either a monomer or a dimer, but bound to GH-2 and PRL-1D elements as a monomer only. The R271W mutation was predicted to affect Pit-1 dimerization, because this residue makes an important POU[sub]s[/sub] domain contact in the dimer structure. Indeed, the R271W mutant protein bound to the GH-1, PRL-1P and 1D sites as a monomer only, as reported previously. This preserved DNA binding may underlie its ability to act as a dominant-negative mutation, as it would be predicted to block dimer formation on Pit-1 target gene elements. The K216E mutant protein displayed markedly enhanced DNA binding as a dimer on the GH-1 element, which may explain its dominant activity, since K216E would be expected to bind preferentially to DNA over the WT protein in the heterozygous state. However, the K216E mutant protein did not bind to the GH-2 and PRL-1D or PRL-1P elements.[br][bold]Conclusion: [/bold]The results indicate the complexity of DNA binding properties of the mutant proteins on multiple target elements. Differential binding may explain the variable effects of Pit-1 mutations on the expression of target genes reflected in the heterogeneity of biochemical phenotypes of patients with CPHD.[br][br]Nothing to Disclose: SD, FEW, SR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2098 313 2386 MON-497 PO20-01 Monday 2055 2012


2051 ENDO12L_MON-498 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Missense Sequence Variants in [italic]CHD7[/italic] Associated with Hypopituitarism Louise Cheryl Gregory, Mark J McCabe, Evelien Gevers, Joanna Baker, Maria Caimari, Mehul T Dattani UCL Institute of Child Health, London, UK; Great Ormond Street Hospital for Children, London, UK; Kent and Canterbury Hospital, Canturbury, UK; Hospital Universitario Son Espases, Palma de Mallorca, Spain CHARGE syndrome is a multisystem autosomal-dominant disorder consisting of coloboma of the eye, heart malformations, atresia of the choanae, retardation of growth and development, genital abnormalities and ear abnormalities. The ATP-dependent chromatin remodeler, chromodomain helicase DNA binding protein-7 (CHD7), is associated with CHARGE and is mutated in up to 65% of patients. [italic]CHD7[/italic] is essential for forebrain development and is expressed in the pituitary and the developing hypothalamus in early embryogenesis. CHD7 and SOX2 interact in neural development, with the latter being implicated in septo-optic dysplasia. [italic]CHD7[/italic] mutations are usually [italic]de novo[/italic] and in heterozygous form. KS and/or hypopituitarism and CHARGE syndrome are known to have overlapping phenotypes. We hypothesised that mutations in the [italic]CHD7[/italic] gene may be implicated in hypopituitarism. We aimed to screen 102 patients with hypopituitarism, including two patients with additional features of CHARGE, for mutations in [italic]CHD7[/italic]. PCR amplification and direct sequencing analysis were used to screen the gene. We identified two heterozygous sequence variants in [italic]CHD7[/italic] in two unrelated patients with CHARGE syndrome. The first missense variant in exon 4, c.2194C[gt]G, p.P732A, is predicted to be functionally deleterious and was identified in a male with severe short stature (-4.9SDS), Tetralogy of Fallot, hearing loss, micropenis, abnormally shaped ears, a squint and hypermetropia, trigonocephaly, a small occipital meningocele, severe growth hormone deficiency (GHD) and mild secondary hypothyroidism. MRI showed a small anterior pituitary (AP), ectopic posterior pituitary (EPP) and a thin corpus callosum (CC). This variant has previously been described in a patient with CHARGE but was not identified in 1500 controls, nor in an additional 100 CHARGE or hypopituitarism patients. A second intronic variant c.IVS35+6T[gt]C, was identified in a male with a full spectrum of CHARGE features in addition to GHD, GnD, TSHD and ACTHD plus a thin CC, cavum septum pellucidum, EPP and a small AP on MRI. It is predicted to affect splicing and was not detected in 1095 controls nor in any of our other patients. In both cases, the mother was the heterozygous carrier. We present two variants in the [italic]CHD7[/italic] gene, one of which is novel, in two unrelated patients with hypopituitarism and features of CHARGE syndrome. The variants may contribute to hypopituitarism in our patients, but are not causative in isolation.[br][br]Nothing to Disclose: LCG, MJM, EG, JB, MC, MTD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 377 313 2387 MON-498 PO20-01 Monday 2056 2012


2052 ENDO12L_MON-499 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Overexpression of Activated Notch in Pou1F1-Expressing Cells Leads to Mild Defects in the Post-Natal Pituitary Gland Leonard YM Cheung, Robin Lovell-Badge, Karine Rizzoti, Paul Le Tissier National Institute for Medical Research, London, UK The Notch signaling pathway is a conserved signal transduction pathway, acting through ligand binding to the eponymous transmembrane receptors (Notch 1-4 in mammals). In the developing pituitary gland Notch signaling is required for differentiation of several endocrine cell types (1, 2). Moreover, transgenic overexpression of Notch1 in Pou1F1-expressing cells leads to growth retarded mice with loss of differentiated hormone cell types (4), and persistent Notch signalling in other pituitary cell lineages leads to delayed differentiation (5, 6). Notch signaling components are also expressed in the adult pituitary gland, including the putative stem/progenitor cells (7). Our studies have investigated the effects of activated Notch signaling in differentiated cells of the Pou1F1-lineage through persistent expression of activated Notch2.[br]Using isolated growth hormone (GH) and prolactin cell populations, we have found that components of the Notch signaling pathway are expressed in these differentiated cell populations in the adult. Further analysis demonstrates that several components of the Notch pathway are differentially expressed between these closely related cell types, with higher expression in GH cells, suggesting that this signalling pathway may play a role in normal somatotroph function. We have then overexpressed constitutively activated Notch2 in somatotrophs using GH-Cre; ROSA26-NICD2 animals but found no dramatic effects on normal growth. Earlier activation from e14.5 in the Pou1F1 lineage using Pou1F1-Cre transgenic mice does not dramatically affect the morphology of the pituitary of newborn animals, or the weight of transgenic mice from juveniles (3-weeks-old) to adults (14-weeks-old). Using radioimmunoassay to measure total pituitary hormone content, we find a mild reduction in GH content in both males and females, as well as minor effects on prolactin and TSH content. This is in contrast to previous studies with activation of Notch signaling in the Pou1F1 lineage resulting in severe growth retardation as a result of failure of GH cell differentiation. The difference in phenotype may result from a later activation of Notch signaling in our model or a super-activation in the previous studies. Our studies suggest that persistent Notch expression in Pou1F1-expressing cells does not have a dramatic effect on the post-natal proliferation or function of differentiated cells of this lineage.[br][br](1) Akimoto et al. Cell Tissue Res. 2010; 340:509. (2) Raetzman et al. Dev Biol. 2007; 304: 455. (3) Raetzman et al. Dev Biol 2004; 265: 329. (4) Zhu et al. Genes Dev. 2006; 20: 2739. (5) Raetzman et al. Mol Endocrinol. 2006; 20: 2898. (6) Goldberg et al. Dev Biol. 2011; 358; 23. (7) Chen et al. Mol Endocrinol. 2006; 20: 3293.[br][br]Nothing to Disclose: LYMC, RL-B, KR, PLT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 978 313 2388 MON-499 PO20-01 Monday 2057 2012


2053 ENDO12L_MON-500 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Three-Dimensional Studies of [italic]Prop1[/italic]-Expressing Cells in the Rat Pituitary Just before Birth Hideji Yako, Takako Kato, Saishu Yoshida, Yukio Kato, Naoko Kanno Graduate School of Agriculture, Meiji University, Kawasaki, Japan; Reproduction and Endocrinology, Kawasaki, Japan; School of Agriculture, Meiji University, Kawasaki, Japan The pituitary lobe is composed of the anterior, intermediate and posterior lobes. The embryonic anterior pituitary progresses daily to differentiate into five types of hormone-producing cells and some types of non-hormone producing cells under spatio-temporal regulation by signal molecules and transcription factors. Recently we demonstrated that progression of differentiation starts in the lateral region of the rat pituitary primordium of embryonic day (E) 13.5 (Yako[italic] et al.[/italic] 2011).[br]In this study, we analyzed heterogeneous property of the cells in the rat pituitary of E21.5, just before birth, by immunohistochemistry using antibodies against three marker proteins (SOX2; marker of stem/progenitor cell, PROP1 (Prophet of Pit1); pituitary-specific transcription factor of heritable gene for combined pituitary hormone deficiency (CPHD), PRX (PRX1 and PRX2); paired-related homeodomain proteins which were identified as novel transcription factor of the pituitary.[br]Three-dimensional quantification of in the anterior pituitary lobe of E21.5 shows an unequal distribution of developing cells in reference to three marker proteins. In the parenchyma, the number of cells positive to PROP1, PRX or SOX2 was larger than in lateral region than in medial region. They are consisting of SOX2-expressing stem/progenitor cells (PROP1[sup]-[/sup]/PRX[sup]-[/sup]/SOX2[sup]+[/sup]; SOX2-only, PROP1[sup]+[/sup]/PRX[sup]-[/sup]/SOX2[sup]+[/sup], PROP1[sup]-[/sup]/PRX[sup]+[/sup]/SOX2[sup]+[/sup] and PROP1[sup]+[/sup]/PRX[sup]+[/sup]/SOX2[sup]+[/sup]) and cells only expressing PRX (PROP1[sup]-[/sup]/PRX[sup]+[/sup]/SOX2[sup]-[/sup]). Triple immunostaining for PRX, vascular endothelial cell marker PECAM and LAMININ demonstrated that most of the PRX-only cells are not pituitary origin but are associated with vasculogenesis in the anterior pituitary. On the other hand, cells lining marginal cell layer were mostly the stem/progenitor cells, while PRX-only cells were absent.[br]In conclusion, the present study indicates that differentiation of cells progresses unequally faster in the lateral and dorsal regions than medial and ventral regions in the parenchyma and vasculogenesis (PRX-only cells) progresses faster in the lateral region than in the medial region.[br][br]Yako H et al., Cell and Tissue Research, 346: 339-346. 2011.[br][br]Nothing to Disclose: HY, TK, SY, YK, NK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1351 313 2389 MON-500 PO20-01 Monday 2058 2012


2054 ENDO12L_MON-501 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) The Effect of PRL-Derived Vasoinhibin in Mammary Cell Proliferation and Apoptosis in Mice Tsukasa Watanabe, Takahiro Yoshida, Midori Maehara, Michiyo Ishida, Toshio Harigaya Meiji University, Kawasaki, Japan Prolactin (PRL) is a 23kDa peptide hormone, produced mainly in the anterior pituitary gland, and it is well known to play a key role in the development, differentiation, and milk synthesis during pregnancy and lactation. Furthermore, PRL has also expressed in non-pituitary tissues including mammary gland (MG). Moreover, N-terminal PRL fragment, vasoinhibins, arises from partial proteolysis of the native PRL has an antagonistic function to native PRL. PRL is angiogenic but vasoinhibin is antiangiogenic. The MG is exocrine gland which secretes milk and undergoes dramatic changes in growth, involution, remodeling, and keep function to satisfy the temporary demands of the young for milk. For example, in the pregnancy, MG is developed for lactation, and mammary epithelium cells (MECs) cause large-scale cell proliferation. At the weaning, MECs are going into apoptosis and MG returns to the virgin stage. From these backgrounds, we hypothesize that the intact PRL is contributed to angiogenesis on the pregnant and lactating MG and vasoinhibin contributed to anti-angiogenesis on MG weaning.[br]In the present study, we detected the results as follows; (1)Vasoinhibin was incorporated into day 16 pregnant MG fragments in vitro. (2)Cyclin D1 mRNA expression was decreased in day 16 pregnant MG fragments by vasoinhibin. (3)Apoptotic positive cells were increased in day 16 pregnant MG fragments by vasoinhibin. (4) Apoptotic positive cells were significantly increased in human breast cancer cell line (Michigan Cancer Foundation-7:MCF7) by vasoinhibin.[br]These results suggested that vasoinhibin was incorporated into MG in tissue culture. This incorporated vasoinhibin involved to effect directly on MECs and then increased apoptosis positive cells in pregnant mouse MG and MCF7. It was also suggested that the vasoinhibin suppressed the cell growth of MECs.[br][br]Nothing to Disclose: TW, TY, MM, MI, TH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1335 313 2390 MON-501 PO20-01 Monday 2059 2012


2055 ENDO12L_MON-502 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) The Natural History of Macroprolactinemia Naoki Hattori, Takashi Ishihara, Akira Shimatsu Ritsumeikan University, Kusatsu, Japan; Kobe City General Hospital, Kobe, Japan; National Hospital Organization Kyoto Medical Center, Kyoto, Japan Macroprolactinaemia is a condition where macroprolactin (MW[gt]150kDa) is predominantly present in the sera. It is one of the causes of hyperprolactinaemia and the prevalence is reportedly 10-25% in patients with hyperprolactinaemia. Macroprolactin is mainly a complex of PRL with immunoglobulin G (IgG), especially anti-PRL autoantibodies. The natural history of macroprolactinaemia is currently unknown. It is unclear when macroprolactinaemia develops in life and whether the PRL status returns to normal later in life. It is also unclear whether individuals with macroprolactinaemia and normal initial serum PRL levels (normoprolactinaemia) eventually develop hyperprolactinaemia. The aim of this study was to examine the natural history of macroprolactinaemia.[br][bold]Subjects and methods[/bold]: Six hundred and fifty-four hospital workers (520 women and 134 men, aged 37.4 [plusmn] 9.7 years) participated, including 27 subjects with macroprolactinaemia (21 women and 6 men) and 627 control subjects (499 women and 128 men). Macroprolactin and serum PRL concentrations were evaluated over a 4-year period. The ratio of macroprolactin was examined by the polyethylene glycol (PEG) method and gel filtration chromatography. IgG-bound PRL and anti-PRL autoantibodies were examined by protein G and [sup]125[/sup]I-PRL binding studies, respectively.[br][bold]Results[/bold]: Over the 4 years of the study all 27 macroprolactinaemic subjects had persistent macroprolactinaemia, while none of the 627 control subjects developed macroprolactinaemia. In subjects with macroprolactinaemia, the ratios of PEG-precipitable PRL and IgG-bound PRL did not significantly change, but [sup]125[/sup]I-PRL binding ratios significantly increased. The number of macroprolactinaemic subjects, whose sera contained anti-PRL autoantibodies, increased from 13 to 21. As a whole, total and free serum PRL concentrations did not significantly change over the 4-year period. However, hyperprolactinaemia developed in 5 of the 18 macroprolactinaemic subjects who were initially normoprolactinaemic along with an increase in anti-PRL autoantibody titres. One of the 9 macroprolactinaemic subjects who were initially hyperprolactinaemic showed a decrease in serum PRL concentrations, which occurred concomitantly with a decrease in the anti-PRL autoantibody titre.[br][bold]Conclusions[/bold]: Macroprolactinaemia may develop prior to middle age and is likely a chronic condition leading to hyperprolactinaemia.[br][br]Nothing to Disclose: NH, TI, AS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 162 313 2391 MON-502 PO20-01 Monday 2060 2012


2056 ENDO12L_MON-503 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Novel Gene Product of TAC3 in Somatolactin and Prolactin Regulation in Grass Carp Pituitary Cells: Receptor Specificity and Signal Transductions Guangfu Hu, Mulan He, Wendy KW Ko, Anderson OL Wong The University of Hong Kong, Hong Kong, Hong Kong TAC3 is a member of tachykinin gene family and its gene product neurokinin B (NKB) has recently emerged as a key factor regulating reproductive function via the kisspeptin/GnRH system. Unlike mammals, the biological functions of NKB in lower vertebrates have not been characterized. Using grass carp as an animal model, we sought to examine the regulatory actions of TAC3 gene products at the pituitary level in modern day bony fish. As a first step, the structural identity of grass carp TAC3 was established by molecular cloning. Unlike the TAC3 genes in mammal and birds, which encode a single mature peptide for NKB, the carp TAC3 encodes two mature peptides, including NKB and a novel peptide that has not been reported previously. Since the novel peptide is structurally related to NKB, it was named NKB-related peptide (NKBRP). In grass carp, TAC3 gene was found to be expressed mainly in the hypothalamus, and to a lower extent in the pituitary. At the pituitary level, TAC3 transcript was detected in somatotrophs and lactotrophs but not in gonadotrophs. In carp pituitary cells, NKB and NKBRP were both effective in stimulating prolactin (PRL) and somatolactin [alpha] (SL[alpha]) secretion and mRNA expression. Using receptor-specific agonists and antagonists for different tachykinin receptor subtypes, we have demonstrated for the first time that the stimulation effects of the two TAC3 gene products on PRL and SL[alpha] secretion and mRNA expression were mediated via pituitary NK2 receptor (NK2R) and NK3 receptor (NK3R), respectively. Using a pharmacological approach, NK3R activation by carp NKB and NKBRP could induce SL[alpha] secretion and mRNA expression via the cAMP/PKA and PLC/IP3/PKC pathways and secondarily coupling of Ca[sup]2+[/sup]/CaM/CaM K-II cascades. Apparently, the Ca[sup]2+[/sup]-dependent mechanisms could be initiated by two functional components, namely [Ca[sup]2+[/sup]][sub]e[/sub] entry via L-type Ca[sup]2+[/sup] channels (VSCC) and [Ca[sup]2+[/sup]][sub]i[/sub] mobilization of IP3-sensitive Ca[sup]2+ [/sup]stores. In the case of PRL release and gene expression, NK2R activation at the pituitary level could also activate cAMP/PKA, PLC/IP[sub]3[/sub] and Ca[sup]2+[/sup]/CaM/CaM K-II cascades, but PKC was not involved. These results, as a whole, demonstrated for the first time that similar to NKB, NKBRP, the novel gene product of carp TAC3, could induce SL[alpha] and PRL expression in the carp pituitary via different tachykinin receptor subtypes couple to overlapping and yet distinct signal transduction mechanisms.[br][br]Nothing to Disclose: GH, MH, WKWK, AOLW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 338 313 2392 MON-503 PO20-01 Monday 2061 2012


2057 ENDO12L_MON-504 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Regulatory Role of Melatonin and BMP-4 in Prolactin Secretion by Rat Pituitary Lactotrope Cells Kanako Ogura-Ochi, Fumio Otsuka, Naoko Tsukamoto, Tomoko Miyoshi, Eri Nakamura, Masaya Takeda, Kenichi Inagaki, Toshio Ogura, Yasumasa Iwasaki, Hirofumi Makino Okayama University Graduate School, Okayama, Japan; Kochi University, Nankoku, Japan Melatonin (N-acetyl-5-methoxytryptamine) is secreted from the pineal gland. It has been recognized that increase in melatonin at night plays a key role as biological clock, which also affects immune system, antioxidant actions, and secretion of various hormones and growth factors. However, the effect of melatonin on prolactin secretion and the regulatory mechanism in the anterior pituitary remain uncertain. We here studied the regulatory role of melatonin in prolactin production using rat pituitary lactotrope GH3 cells by focusing on the pituitary bone morphogenetic protein (BMP) system, which has been known to induce prolactin production and lactotrope cell proliferation. In GH3 cells, melatonin MT2 receptor was predominantly expressed compared with MT1 receptor. Melatonin and its analog, ramelteon, significantly suppressed basal as well as forskolin-induced prolactin secretion and the level of prolactin mRNA expression in a concentration-responsive manner with reduction of cAMP synthesis. Of note, the inhibitory effects of melatonin and ramelteon were reversed by treatment with a selective MT2 receptor antagonist, luzindole, suggesting functional involvement of MT2 action in the suppression of prolactin release. Moreover, melatonin and ramelteon also inhibited BMP-4-induced prolactin secretion and its mRNA expression by inhibiting BMP-induced Smad1/5/8 phosphorylation and the transcription of BMP-target gene Id-1 partly through the activation of AKT signaling pathway. It was thus suggested that melatonin MT2 activity also acts as an inhibitor for prolactin secretion through suppression of BMP-4 actions in lactotrope cells. Considering that BMP-4 increased the expression levels of MT2 mRNA in lactotrope cells, the sensitivity for endogenous melatonin could also be upregulated by BMP-4. Collectively, a regulatory interaction of melatonin and BMP-4 in prolactin production was uncovered in pituitary lactotorope cells. Attempt to upregulate MT2 bioavailability may be clinically applicable for the treatment of pituitary prolactinomas when the tumors are resistant to dopamine agonists.[br][br]Nothing to Disclose: KO-O, FO, NT, TM, EN, MT, KI, TO, YI, HM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1144 313 2393 MON-504 PO20-01 Monday 2062 2012


2058 ENDO12L_MON-505 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Characterization of Lactotrope Intracellular Calcium Response to TRH and Dopamine in Acute Pituitary Slices from Lactating and Cycling Mouse Luis Miguel Rendon, Tatiana Fiordelisio Universidad Nacional Aut[oacute]noma de M[eacute]xico, Mexico City, DF, Mexico Hypothalamic TRH and/or DA trigger or inhibit lactotrophs to synthesize and secrete prolactin, hormone that has an important role in lactogenic activity, lipidic metabolism and some effects in the immune system. Different doses of TRH produce characteristic signalling patterns of intracellular calcium [Ca2+]i. It has been proposed before that lactotrophs respond to stimuli in an heterogeneous way, this variation depends on its localization in the pituitary. However, these studies were done in primary culture conditions, where the cells lose their interaction with others and their natural physiologic environment. With the main purpose of determining if a physiological plasticity exists in these cells that allows to cover the changes in the demand of hormonal secretion, we use acute pituitary slices from lactating and cycling mouse and intracellular Ca2+ imaging to examine the spatial distribution of lactotrophs and their Ca2+ signaling patterns elicited by different TRH concentrations and dopamine.[br]Adult female Balb-C mouse were euthanized. Pituitary gland was removed and embedded in agar 3% and coronal slices of 130 [micro]m of thickness were obtained. Slices were incubated with fluo-4 AM with saline saturated with 95% O2 and 5% CO2. Image sequences at 510 nm emission were obtained with a stereomicroscope and cooled camera CCD. TRH was bath-applied for 30s at concentrations 0.1 nM to 100 nM. Finally, separate dopamine 2 [mu]M and high potassium solution, to evaluate viability, were applied.[br]The responsive cells to TRH and DA were counted in two different regions, the central region and lateral region of the gland and their patterns of response of [Ca2+]I were plotted and analyzed individually.[br]With this protocol we found that in the central region, like in the lateral region of the pituitary gland, the number of responding cells increases proportionally to doses of TRH, being major at 100 nM. Moreover, the number of cells with [Ca2+]i activity in response to TRH is minor in lateral region in comparison with the central region. We found that the lactotrophs [Ca2+]I response produced by TRH present higher amplitude in cells in the central region of hyphophysis. Finally, in comparison with previous studies in males, we find that in female, the lactotrophs show a more homogeneous distribution in the lobe, suggesting increased cell proliferation and plasticity of the pituitary gland because of physiological differences in lactation and oestrus cycle.[br][br]Nothing to Disclose: LMR, TF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2093 313 2394 MON-505 PO20-01 Monday 2063 2012


2059 ENDO12L_MON-506 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Haloperidol-Induced Hyperprolactinemia Counteracts Joint Inflammation in Adjuvant-Induced Arthritis in Rats Guadalupe Ledesma-Colunga, Norma Adan, Andres Quintanar-Stephano, Sthepanie Thebault, Gonzalo Martinez de la Escalera, Carmen Clapp Universidad Nacional Aut[oacute]noma de M[eacute]xico, UNAM, Quer[eacute]taro, Mexico; Universidad Aut[oacute]noma de Aguascalientes, Aguascalientes, Mexico Prolactin (PRL) may play a protective role against the progression of rheumatoid arthritis (RA), an autoimmune disease characterized by joint inflammation. PRL exerts immunoregulatory effects, and it is elevated in the circulation of patients with RA. Moreover, breast-feeding, which is accompanied by high circulating PRL levels, can reduce the risk of RA. Here, we investigated the effect of hyperprolactinemia on joint inflammation by using the adjuvant-induced RA model in rats. The subcutaneous placement of tablets containing haloperidol, a dopaminergic D2 antagonist resulting in the pituitary secretion of PRL, was used to induce hyperprolactinemia. Haloperidol treatment initiated 3 days before the injection of complete Freund[apos]s adjuvant (CFA) counteracted arthritis development, evaluated by the progressive increase in hind paw swelling, pain, expression of proinflammatory mediators (iNOS, TNF-[alpha], IL-1[beta], IFN-[gamma], IL-6, MMP-3, MMP-9, MMP-13), and the reduction of body weight occurring within 10 to 21 days after inoculation. Haloperidol produced hyperprolactinemia only during the first 14 days after treatment, suggesting that high levels of circulating PRL during the early stages of CFA-induced arthritis are enough to protect against the progression of the disease for at least 21 days. In support of PRL mediating haloperidol protection, chronic hyperprolactinemia induced by placing two anterior pituitary glands under the kidney capsule prevented CFA-induced hind paw pain and proinflammatory mediator expression. This protection was blocked by the D2 agonist cabergoline, which lowered the levels of circulating PRL. These findings support haloperidol-mediated hyperprolactinemia as a promising therapeutic strategy for controlling the progression of RA.[br][br]Sources of Research Support: UNAM grant IN200312.[br][br]Nothing to Disclose: GL-C, NA, AQ-S, ST, GMdlE, CC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 866 313 2395 MON-506 PO20-01 Monday 2064 2012


2060 ENDO12L_MON-507 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Prolactin Promotes Differentiation of Breast Cancer Cells through Suppression of CK5 Takahiro Sato, Lynn M Neilson, Hallgeir Rui Thomas Jefferson University, Philadelphia, PA Prolactin (PRL) is a critical regulator of normal growth, development, and differentiation of breast epithelia. While prolactin has been shown to promote mammary tumorigenesis, accumulating evidence also supports a role for PRL in maintaining breast cancer cell differentiation. A principal signaling mediator of PRL, Stat5, promotes differentiation of human breast cancer cells [italic]in vitro[/italic] (1). In addition, active Stat5 is an independent favorable prognostic marker, and loss of active Stat5 is associated with increased risk of tamoxifen therapy failure in breast cancer patients (2). Progesterone (PG) has recently been shown to increase a basal-like CK5 positive population in luminal breast cancer cell lines. These PG-induced CK5-positive cell populations are quiescent, therapy-resistant, and have tumor-initiating capability, indicating that they represent a progenitor/cancer stem cell population (3). We now report that PRL can counteract the PG induced increase in the CK5 population in luminal breast cancer cells. PRL suppresses the population of CK5 positive cells both [italic]in vitro[/italic] and [italic]in vivo[/italic]. We further report that PG is a potent inducer of BCL6, and that BCL6 induction was required for PG-induction of CK5 positive cells based on BCL6 shRNA-mediated knockdown. BCL6 has been shown to be critical for the inhibition of differentiation of germinal center B-cells and for the maintenance of leukemia initiating cells (4). We and others have also shown that basal levels of BCL6 are rapidly suppressed by PRL in breast cancer cells, and BCL6 is expressed during early pregnancy, when other reproductive hormones such as PG and Estrogen are elevated (5, 6). We now report that PRL is capable of suppressing not only basal levels of BCL6 but also the robust induction of BCL6 by PG. We propose that PRL prevents PG-induced CK5 positive progenitor cell expansion through a mechanism that involves suppression of PG-induced BCL6. These opposing effects of PRL and PG may have important implications for drug responsiveness of breast cancer.[br][br](1) Sultan, A.S., et al., Stat5 promotes homotypic adhesion and inhibits invasive characteristics of human breast cancer cells. Oncogene, 2005. 24(5): p. 746-60. (2) Peck, A.R., et al., Loss of nuclear localized and tyrosine phosphorylated Stat5 in breast cancer predicts poor clinical outcome and increased risk of antiestrogen therapy failure. J Clin Oncol, 2011. 29(18): p. 2448-58. (3) Kabos, P., et al., Cytokeratin 5 positive cells represent a steroid receptor negative and therapy resistant subpopulation in luminal breast cancers. Breast Cancer Res Treat, 2011. 128(1): p. 45-55. (4) Duy, C., et al., BCL6 enables Ph+ acute lymphoblastic leukaemia cells to survive BCR-ABL1 kinase inhibition. Nature, 2011. 473(7347): p. 384-8. (5) Tran, T.H., et al., Prolactin inhibits BCL6 expression in breast cancer through a Stat5a-dependent mechanism. Cancer Res, 2010. 70(4): p. 1711-21. (6) Logarajah, S., et al., BCL-6 is expressed in breast cancer and prevents mammary epithelial differentiation. Oncogene, 2003. 22(36): p. 5572-8.[br][br]Nothing to Disclose: TS, LMN, HR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1689 313 2396 MON-507 PO20-01 Monday 2065 2012


2061 ENDO12L_MON-508 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) The Post-Pubertal Decline in Pulsatile Growth Hormone Secretion in Mice Is Exacerbated by Increased Adiposity Following High-Fat Feeding Lili Huang, Frederik J Steyn, Hwee Y Tan, Terasa Y Xie, Johannaes D Veldhuis, Chen Chen University of Queensland, Brisbane, Australia; Mayo Clinic, Rochester, MN The transition from puberty to adulthood coincides with a reduction in circulating levels of growth hormone (GH). A pathological decline in post-pubertal GH secretion is detrimental to attainment of peak lean muscle and bone mass, and promotes adiposity and susceptibility for the development of obesity and diabetes. It is thus essential that we identify the factors that may advance the post-pubertal decline in GH secretion.[br]To characterize the impact of dietary induced weight gain and increased adiposity on the post-pubertal decline of GH secretion, we first characterized pulsatile GH secretion in C57Bl/6J mice maintained on a standard diet at 12 and 16 weeks of age. We then assessed pulsatile GH secretion in mice at 12 weeks of age following 4 weeks of high fat feeding. In addition, circulating levels of leptin, insulin, glucose, free fatty acids (FFAs) and triglycerides were assessed.[br]We observed a significant decline in pulsatile GH secretion in mice from 12 to 16 weeks of age. This was characterized by a significant decline in total (725[plusmn]136 vs 365[plusmn]68.3ng/ml per 6h, p[lt]0.05), pulsatile (594[plusmn]129 vs 326[plusmn]68.5ng/ml per 6h, p[lt]0.05), and the mass of GH secreted per burst (143[plusmn]17.0 vs 101[plusmn]24.2ng/ml, p[lt]0.05). High fat feeding resulted in a significant increase in adiposity (as reflected by increased gonadal fat mass (0.40[plusmn]0.05 vs 1.21[plusmn]0.15g; p[lt]0.05) and leptin levels (5.24[plusmn]0.88 vs 22.1[plusmn]1.95ng/ml, p[lt]0.05)), and the development of glucose intolerance and hyperinsulinemia (0.74[plusmn]0.20 vs 1.82[plusmn]0.35ng/ml, p[lt]0.05). The increased adiposity correlated with a significant decline in total (873[plusmn]160 vs 442[plusmn]36.3ng/ml per 6h, p[lt]0.05), pulsatile (732[plusmn]140 vs 414[plusmn]38.8ng/ml per 6h, p[lt]0.05) and basal GH secretion (141[plusmn]32.4 vs 26.7[plusmn]6.02ng/ml per 6h, p[lt]0.05) by 12 weeks of age, similar levels as that in 16 weeks of age on a standard diet. In addition, impaired GH secretion following an increase in adiposity coincided with an elevation in circulating levels of insulin and developing glucose intolerance but not an elevation in circulating levels of FFAs.[br]Observations suggest that dietary induced weight gain and increased adiposity is associated with an impairment of pulsatile GH secretion in mice, and the advance in post-pubertal decline in GH secretion. Given the established role of GH on post-pubertal somatic development, we anticipate that reduced GH secretion as a consequence of dietary induced weight gain and increased adiposity will have long-term ramifications on adult health.[br][br]Sources of Research Support: This work was supported by the Australian National Health and Medical Research Council (NHMRC) and The University of Queensland. L. Huang receives an overseas postgraduate research scholarship from China Scholarship Council (CSC) and subsidy scholarship from University of Queensland.[br][br]Nothing to Disclose: LH, FJS, HYT, TYX, JDV, CC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 884 313 2397 MON-508 PO20-01 Monday 2066 2012


2062 ENDO12L_MON-509 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Development of a Tissue-Specific Leptin Knockout Mouse: Validation and Unexpected Challenges Angela Katherine Odle, Melody Lyn Allensworth-James, Anessa C Haney, Noor Akhter, Gwen Vaughn Childs University of Arkansas for Medical Sciences, Little Rock, AR Thought to primarily be an adipokine, leptin is known to regulate appetite and energy expenditure via receptors in the hypothalamus. However, researchers now report extra-adipocyte sources of leptin with the potential for additional functions such as paracrine or developmental regulation. One of these sources is the pituitary somatotrope and, in order to determine a function for somatotrope leptin, we developed the first floxed leptin model with the use of Cre-loxP technology. Exon 3 of the Lep gene was targeted in a floxed leptin mouse line (FVB-P129). To target pituitary somatotropes, we crossed homozygous floxed leptin (Lepfl/fl) mice with a line of mice in which Cre expression is driven by the rat growth hormone promoter (rGHp-cre, Luque et al, 2007). To validate the functionality of the knockout, we used an adenovirus promotor-driven line to essentially produce a global knockout of leptin (FVB/N-Tg(EIIa-Cre)C5397Lmgd/J; Jackson Laboratory).[br]Extensive genotyping has been performed on the Lepfl/fl, Cre-GH+/- deletion mutants and Lepfl/fl controls to assess the specificity of the knockout. Sixteen tissue samples were collected from F2 and F3 generation homozygous deletion mutants and controls in the Cre-GH, Lepfl/fl line. Unexpected extra-pituitary excision was found in several organs of Cre+ mice, including fat. About 55% of the F3 generation had either one or both alleles of floxed leptin excised in tail snips. To test this further, the line was regenerated using fresh Lepfl/fl founders and fresh Cre-GH mice. Using this system, the regenerated F2 generation has shown only 32% extra-pituitary excision, which suggests that breeding may reduce the non-selective Cre-recombinase expression. As the Cre-GH line has been used extensively without evidence of any extra-pituitary Cre-Recombinase protein production (Luque et al, 2007) or gene excision (Childs et al, 2011), we hypothesize that the floxed leptin gene carried by our mice is very easily excised and may have been the target of extra-pituitary GH (Harvey and Hull, 1977). To get a more selective knockout, we are currently crossing the Lepfl/fl mice with mice in which Cre expression is under the control of the rat growth hormone releasing hormone receptor promoter (FVB(Cg)-Tg(Ghrhr-cre)3242Lsk/J, Yin et al, 2008). Because of the different susceptibilities of floxed genes to Cre excision, these studies emphasize the importance of organ genotyping to check selectivity of Cre-recombinase expression.[br][br]Nothing to Disclose: AKO, MLA-J, ACH, NA, GVC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2256 313 2398 MON-509 PO20-01 Monday 2067 2012


2063 ENDO12L_MON-510 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Neuronal Marker Expression (NF68) in Anterior Pituitary Cells from Lactating, Adrenalectomized and Orchidectomized Rats Avelino Guardado, Tatiana Fiordelisio Facultad de Ciencias, UNAM, Mexico City, Mexico The anterior pituitary (AP) is responsible for systemic hormonal regulation and requires a complex regulation system which integrates signals from different origins, including hypothalamic releasing and inhibiting factors, hormonal factors from target organs, neuronal factors (GABA, dopamine, noradrenaline, etc), and factors derived from the AP itself by means of autocrine and paracrine secretion. In response to all these factors, pituitary secreting cells react with a series of changes, like cytoskeletal remodeling, which control hormone synthesis and release. It has been observed that intermediate filaments regulate the arrangement of other cytoskeletal elements in several cell types, and that they serve as an anchoring site for a great number of enzymes. Neurofilaments (NFs) are intermediate filament proteins from the neuronal cytoskeleton that help determine axonal caliber and influence the speed of vesicular transport. Originally considered exclusive of neuronal cells, NFs have been observed in subpopulations of somatotrophs, lactotrophs, gonadotrophs and tyrotrophs in adenohypophysis. In order to study the possible relation of NFs in the regulation of AP secretion, we have performed double inmunostainings of primary cell cultures of two months old male and female rat adenohypophysis which were adrenalectomized, castrated, or lactating, in order to provoke a physiological demand for certain hormones. All females were in estrous. Monoclonal mouse anti-NF68 and the F(ab[apos]) fragment of mouse anti-IgG cyanine 5 (Cy5) conjugated was used for NFs, and rabbit IgG anti-[beta]FSH, anti-PRL, anti-[beta]TSH, anti-GH and the F(ab[apos]) fragment of rabbit anti-IgG fluorescein isothiocyanate (FITC) conjugated for hormonal inmunostaining. Image acquisition was done with a CCD camera under fixed parameters for each condition. Analysis was made by cell counts identifying the total number of hormone- and NF-positive cells, as well as double positive and double negative cells, comparing data obtained in different hormones, treatments and sexes. Our results show an increase in NF expression in response to treatment (except in the case of adrenalectomies and corticotroph NF expression) represented by an increase in the percentage of double positives in the sample. This increase may be related with a way of controlling secretion by means of cytoskeletal reorganization (a role described for NFs in neurons) or by regulating ion channel function (a role also proposed for NFs in neurons).[br][br]Nothing to Disclose: AG, TF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 775 313 2399 MON-510 PO20-01 Monday 2068 2012


2064 ENDO12L_MON-511 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Negative Regulation of Neuromedin U mRNA Expression in the Rat Pars Tuberalis by Melatonin Administration Sayaka Aizawa, Mai Nagasaka, Takafumi Sakai, Ichiro Sakata Saitama University, Saitama, Japan NeuromedinU (NMU) is a gut-brain peptide hormone containing 25 amino-acid residues in humans and 23 amino-acid residues in mice and rats. NMU produced in the gastrointestinal tract is involved in the regulation of smooth-muscle contractions, local blood flow, and ion transport in the gut. In addition, intracerebroventricular injection of NMU decreases food intake, along with a concomitant transient increase in core body temperature, gross motor activity, and oxygen consumption. In rats, NMU is highly expressed in the pars tuberalis (PT), which is part of the anterior pituitary gland surrounding the median eminence as a thin cell layer. Although central NMU is believed to play a physiologically important role in energy homeostasis, the regulatory mechanism of NMU mRNA expression in the PT is still unclear. Therefore, in this study, we examined the regulation of NMU mRNA expression in the rat PT.[br]We used male Wistar rats, 8 weeks of age and housed in 12-h light and 12-h dark conditions, and collected tissues in the light phase (Zeitgeber time [ZT] 6) and dark phase (ZT18). To examine the diurnal expression of NMU, in situ hybridization (ISH) and quantitative PCR (qPCR) were performed. ISH showed that NMU mRNA-expressing cells were widely distributed throughout the PT. NMU mRNA expression in the PT showed a circadian rhythm, with the level of NMU mRNA expression approximately 2-times higher at ZT6 than at ZT18. Because it has been reported that [alpha]GSU and TSH[beta] mRNA expression in the PT is regulated by melatonin through the melatonin receptor type 1 (MT1), we hypothesized that melatonin also may have an effect on NMU mRNA expression in the PT. For this experiment, melatonin tube was implanted under the skin for 7 days. Melatonin administration suppressed NMU mRNA expression by 80% in the PT at ZT6. Next, we examined NMU receptor 1 (NMUR1) and 2 (NMUR2) mRNA expression by qPCR, which showed that NMUR2 mRNA was expressed in the central nervous system. Significant levels of NMUR2 mRNA were found in the third ventricle ependymal cell layer (EC), followed by the arcuate nucleus (Arc) and the SC. These results suggest that NMU mRNA expression in the rat PT is negatively regulated by melatonin via MT1, possibly by inducing diurnal rhythmic expression of NMU mRNA. Because the PT is located at the base of the brain and bathed in cerebrospinal fluid (CSF), it may be suggested that NMU produced in the PT is secreted into the CSF, and it acts on NMUR2 in the EC, Arc, and SC.[br][br]Nothing to Disclose: SA, MN, TS, IS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 643 313 2400 MON-511 PO20-01 Monday 2069 2012


2065 ENDO12L_MON-512 POSTER SESSION: Pituitary Hormones [amp] Development (1:30 PM-3:30 PM) Stimulatory Regulation of Thyroid-Stimulating Hormone (TSH) [beta] Subunit mRNA Expression and TSH Production by Glutamine and Glutamic Acid in the Rat Pars Tuberalis Ichiro Sakata, Sayaka Aizawa, Mai Nagasaka, Takafumi Sakai Saitama University, Saitama, Japan The pars tuberalis (PT) is a part of the anterior pituitary gland, surrounding the median eminence as a thin cell layer. Studies have shown that most of the hormone-producing cells in the rat PT are TSH-producing cells (PT-TSH cells), which express high levels of melatonin receptor type 1 (MT1). We have previously shown that TSH[beta] and [alpha]GSU mRNA expression in the PT exhibits diurnal variations and is significantly suppressed by melatonin administration, suggesting that PT-TSH plays a physiologically important role as a photoperiodic mediator. However, the regulation mechanism of PT-TSH expression and secretion, especially the role of stimulatory regulation factors, is still obscure. We performed a whole-genome expression analysis of PT tissues in an attempt to identify factors that may be involved in the regulation of PT-TSH.[br]We used male Wister rats, 8 weeks of age and housed in 12-h light and 12-h dark conditions, and collected tissues at zeitgeber time 6. Because the PT tissue is too small to remove manually, laser microdissection was performed to collect the PT; mRNA expression was analyzed using the DNA microarray method. The microarray data showed high mRNA expression levels of KA2, which is an ionotropic glutamic acid receptor (iGluR). The level of KA2 mRNA expression was higher than that of MT1 mRNA expression. This observation was confirmed by qPCR and in situ hybridization (ISH). Because it has been reported that glutamic acid is locally synthesized and acts on glutamic acid receptors, we next measured the expression levels of factors involved in glutamic acid synthesis by qPCR. Amino acid transporter A2, glutaminase (Gls), and Gls2 mRNA was highly expressed in the PT as compared to that in the PD. Thus, we examined the effects of glutamine and glutamic acid on TSH[beta] mRNA expression in the PT by using slice cultures and ISH; 2-h and 4-h treatment with L-glutamine and L-glutamic acid significantly stimulated TSH[beta] expression in PT slices, but the effect of L-glutamic acid was faster and stronger than that of L-glutamine. TSH secretion from the PT into the medium was also stimulated by glutamic acid. In contrast, treatment with glutamine and glutamic acid did not alter [alpha]GSU mRNA expression in the PT, or TSH[beta] and [alpha]GSU mRNA expression in the PD. These results suggest that glutamine taken up by PT cells is converted to glutamic acid by Gls, finally inducing TSH[beta] mRNA expression and TSH production through KA2 in an autocrine and/or paracrine manner.[br][br]Nothing to Disclose: IS, SA, MN, TS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 746 313 2401 MON-512 PO20-01 Monday 2070 2012


2066 ENDO12L_MON-524 POSTER SESSION: Nuclear Receptors: Coactivators [amp] Corepressors (1:30 PM-3:30 PM) Effect of Aerobic Exercise on Leptin and PGC-1[alpha] Gene Expression in a Unique Cohort of Morbidly Obese Polynesians with Type 2 Diabetes Irum Hayat, David Rowlands, Isabelle Lys, William Sukala, Birinder Cheema, Bernhard Breier, Murray Leikis, Jeremy Krebs, Rachel Page Massey University, Wellington, New Zealand; Charles Darwin University, Darwin, Australia; University of Western Sydney, Penrith South, Australia; Capital and Coast District Health Board, Wellington, New Zealand; Southern Cross University, East Lismore, Australia; Wellington Hospital, Wellington, New Zealand; Charles Darwin University, Auckland, New Zealand Incidence of type 2 diabetes (T2D) and obesity is rapidly escalating in New Zealand, particularly in Polynesians (1). Aerobic (AER) exercise can improve body composition (2), glycaemic control (2), and alter circulating metabolic hormones in individuals with T2D (3). PGC-1[alpha] is a key regulator in energy metabolism, controlling adaptive thermogenesis and glucose/fatty acid metabolism (4,7), and has been implicated in the pathogenesis of T2D (4,5,6,7). We have a unique cohort of morbidly obese Polynesian New Zealanders with T2D that have undergone intensive exercise training in the SPIRIT study (8). We are interested in examining the impact of exercise on energy metabolism in the skeletal muscle (SM) of this unique cohort.We aim to investigate the impact of 16 week AER exercise in SPIRIT study cohort on a) circulating leptin hormone levels and b) SM gene expression of PGC-1 and associated energy metabolism coregulators.[br]9 participants (mean BMI 45.0 [plusmn] 6.5 kg/m2; mean waist circumference 131.9 [plusmn] 13.5 cm) underwent 16 weeks of AER training. Blood and SM samples were obtained at pre (0 weeks) and post (16 weeks) exercise intervention. Blood plasma samples were analysed for circulating levels of leptin. Gene expression of PGC-1[alpha], FABP4, FABP5, PPAR[gamma], UCP2, UCP3 and leptin receptors were investigated in SM samples using Illumina mRNA microarray. ANOVA and posthoc analysis was performed.[br]There were no significant changes in mean serum leptin levels. Changes in SM mRNA expression for PGC-1[alpha] and coregulators were: PGC-1[alpha](1.2 fold increase; p [lt]0.01); PPAR[gamma] (1.1 fold increase; p[lt]0.05); UCP2 (1.3 fold increase; p [lt]0.001), UCP3 (1.2 fold increase; p [lt]0.05) FABP5 (1.2 fold increase; p [lt]0.001) and leptin receptor (1.1 fold increase; p [lt]0.05). There was no statistically significant change in SM mRNA expression of FABP4.[br]Although there was no change in circulating leptin hormone, the SM leptin receptor showed a significant increase in mRNA expression. This could indicate that 16 weeks of AER exercise improves leptin sensitivity in this morbidly obese cohort. The preliminary results demonstrate that AER training has increased the expression of several key regulatory genes involved in energy metabolism. These results support our hypothesis that metabolic adaptation is occurring in the SM of the SPIRIT study participants even though there is no improvement in their glycaemic control.[br][br]1. Ministry of Health, 2008, A portrait of health: key results of the 2006/2007 New Zealand Survey. Wellington. 2. Sigal, RJ et al., Diabetes Care 2004; 27 (10):2518-2539. 3. McMurray RG et al., Sports Med; 2005, 35 (5):393-412. 4. Liang H et al; Adv Physiol Educ; 2006, 3:145-151. 5. Fink, NB et al; J Clin Inves; 2006,116(30): 615-622. 6. Liu C et al; Acta Biochim Biophys Sin; 2011, 1-10. 7. Koves T., et al; Cell Metab; 2008; 7(1):45-56. 8. Sukala, W et al; Eur J App Physiol; 2012; 112:317-325.[br][br]Nothing to Disclose: IH, DR, IL, WS, BC, BB, ML, JK, RP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1341 314 2402 MON-524 PO23-01 Monday 2071 2012


2067 ENDO12L_MON-525 POSTER SESSION: Nuclear Receptors: Coactivators [amp] Corepressors (1:30 PM-3:30 PM) The Silencing Mediator for Retinoic Acid and Thyroid Hormone Receptor (SMRT) Coregulator Utilizes Distinct Domains to Bind to Estrogen Receptor-[alpha] Bound to Agonist or Antagonist Vaishali Chaubal, Margaret C Pace, Tong Gao, Kendall W Nettles, Carolyn L Smith Baylor College of Medicine, Houston, TX; The Scripps Research Institute, Jupiter, FL [underline]S[/underline]ilencing [underline]m[/underline]ediator of [underline]r[/underline]etinoic acid and [underline]t[/underline]hyroid hormone receptor (SMRT) is a transcriptional coregulator that interacts with the ligand binding domain (LBD) of antagonist-bound ER[alpha] via its two C-terminal CoRNR box motifs also termed interacting domains (IDs). An additional CoRNR box motif in the SMRT N-terminus mediates a ligand-independent interaction with the ER[alpha] DNA binding domain (DBD) of ER[alpha] (MCB 30:1434, 2010). SMRT regulates the transcriptional activity of E2-bound ER[alpha], either positively or negatively, in a gene-specific manner. GST pull-down assays demonstrate moderate binding of ER-LBD to the ID region of SMRT. However, the ability of the SMRT[Delta]ID mutant, which lacks both C-terminal CoRNR box motifs, to stimulate ER[alpha]-dependent gene expression suggested that an additional domain within SMRT could promote E2-stimulated ER[alpha] activity. The SMRT-ID domain does not interact with ER[alpha] AF-1. However, full-length SMRT co-immunoprecipitated with the ER[alpha] AF-1 domain and reduced the transcriptional activity of a GAL4 DBD-AF1 fusion protein measured by luciferase reporter assay, indicating a physical interaction of SMRT with the N-terminus of ER[alpha]. Further evidence that SMRT interacts with the AF-1 domain was obtained in a modified mammalian two-hybrid assay in which SMRT promoted the interaction of an isolated AF1 fragment with VP16-LBD. Moreover, the AF-1 domain modulated ligand-dependent SMRT interactions such that VP16 fused to full-length ER[alpha] was stimulated to bind to GAL4 DBD-SMRTaa2059-2352 (the ID region) to a greater extent by 4-hydroxytamoxifen (4HT) than E2 while VP16-LBD binding to this SMRT fusion protein was stimulated by only E2. Thus the AF-1 domain can interact with SMRT and modulate ligand-dependent interactions of SMRT with full-length ER[alpha]. However, the ER[alpha] AF-1 domain was not required for SMRT[Delta]ID to stimulate E2-dependent ER[alpha] transcriptional activity, as this SMRT deletion mutant was able to coactivate GAL4-LBD. In GST pull-down assays E2 but not 4HT stimulated the interaction of GST-LBD with a SMRT fragment encompassing amino acids (aa) 1243-2103, but not with a shorter SMRT aa1589-2103 region. Taken together these results indicate that the aa1243-1589 region of SMRT binds to the ER[alpha] LBD in an E2-dependent manner, and suggests a model in which the transcriptional activity of ER[alpha] interacting with this subdomain of SMRT is stimulated, while 4HT-stimulated interactions with the SMRT ID region result in repression of ER[alpha]-dependent gene expression.[br][br]Nothing to Disclose: VC, MCP, TG, KWN, CLS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2258 314 2403 MON-525 PO23-01 Monday 2072 2012


2068 ENDO12L_MON-526 POSTER SESSION: Nuclear Receptors: Coactivators [amp] Corepressors (1:30 PM-3:30 PM) Regulation of SMRT Coregulator Isoform Expression by Estrogen Receptor Ligands Julia K Blackmore, Vaishali Chaubal, Carolyn L Smith Baylor College of Medicine, Houston, TX The nuclear receptor corepressor (NCoR) and silencing mediator of retinoic acid and thyroid hormone receptor (SMRT) coregulators were identified through their ability to bind to and inhibit the basal activity of unliganded type II nuclear receptors, while more recently we have shown that SMRT can coactivate or corepress estrogen receptor-[alpha] (ER[alpha]) activity in a gene-specific manner. Previously, estradiol (E2) was shown to down-regulate NCoR protein expression via proteasome-dependent degradation ([italic]PNAS[/italic] 102:13153, 2005). The effect of E2 on expression of SMRT protein is unclear, although this steroid reduces SMRT mRNA expression ([italic]Acta Physiologica[/italic] 197:187, 2009). In MCF-7 cells at least two major forms of SMRT are expressed, full length SMRT[alpha] and the SMRT[beta] splice variant which lacks amino acids encoded by exons 2-6 and therefore the majority of the coregulator[apos]s first repression domain. The ability of estrogen and antiestrogen to regulate these proteins[apos] expression was examined in MCF-7 cells. When cultured in media containing stripped FBS, as little as 6h of E2 treatment increased expression of SMRT[beta] but not SMRT[alpha] protein. The selective estrogen receptor modulator (SERM) 4-hydroxytamoxifen (4HT) also increased SMRT[beta] expression to an extent similar to E2 treatment. Depletion of ER[alpha] by siRNA-mediated technology blocked E2 regulation of SMRT[beta] expression. Moreover, the ICI 182,780 pure ER[alpha] antagonist reduced SMRT[beta] protein in MCF-7 cells cultured in estrogen replete serum and blocked E2-induction of this corepressor isoform, indicating an ER[alpha]-dependent effect. Measurement of SMRT mRNA levels by a Panomics QuantiGene Plex 2.0 assay that assesses the expression of all SMRT isoforms indicated SMRT mRNA was decreased by 24h of either E2 or 4HT treatment. The mRNA reduction suggests that E2 and 4HT increase SMRT[beta] protein expression by a post-transcriptional mechanism. Moreover, similar E2 or 4HT induction of SMRT[beta] protein expression was observed for the MDA-MB-134-VI breast cancer cell line, but not for ZR-75-1, BT-474 or MDA-MB-361 breast cancer cells. All these cell lines are ER[alpha] positive implicating additional cell-type specific factors in regulating SMRT[beta] expression by E2 and SERMs. Collectively, these results demonstrate that ER ligands can directly control transcription through regulation of ER transcriptional activity as well as through altering the expression of SMRT and consequently the activity of a larger spectrum of SMRT-regulated transcription factors.[br][br]Sources of Research Support: DK53002.[br][br]Nothing to Disclose: JKB, VC, CLS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1940 314 2404 MON-526 PO23-01 Monday 2073 2012


2069 ENDO12L_MON-527 POSTER SESSION: Nuclear Receptors: Coactivators [amp] Corepressors (1:30 PM-3:30 PM) SMRT Regulates Glucocorticoid Action in Adipocytes Stelios Mantis, Xuan Han, Michael Landeche, Ronald N Cohen University of Chicago, Chicago, IL; University of Chicago, Chicago, IL Increased glucocorticoid action in adipocytes has been linked to systemic metabolic abnormalities including insulin-resistant diabetes, obesity, dyslipidemia, and hypertension. However, circulating levels of glucocorticoids are not clearly altered in patients with the metabolic syndrome. Glucocorticoid action is regulated by the glucocorticoid receptor (GR), a member of the nuclear hormone receptor (NR) superfamily. NRs recruit coactivators in the presence of ligand, and recruit corepressors either in the absence of ligand or in the presence of antagonists. Although multiple studies have suggested that coactivators are important for GR action, the role of corepressors is much less clear. The two most important nuclear receptor corepressors are the silencing mediator of retinoid and thyroid hormone receptors (SMRT) and the nuclear receptor corepressor (NCoR). To investigate the role of corepressors in GR function in the adipocyte, we created SMRT +/- heterozygous knock-out mice. These mice exhibit normal adiposity on a chow diet, but gain excess weight when placed on a high-fat diet. To investigate the role of SMRT in glucocortioid action, we isolated adipose tissue from SMRT +/- and WT mice and treated it with and without dexamethasone. RNA was isolated, and we performed quantitative RT-PCR of the dexamethasone-responsive gene GILZ. Interestingly, dexamethasone-mediated GILZ expression was 50% higher in adipose tissue derived from SMRT +/- mice compared to WT controls. To determine if glucocorticoid-responsive changes in gene expression were due to direct recruitment of SMRT by the GR, we performed chromatin immunoprecipitation (ChIP) experiments, taking advantage of the well-characterized glucocorticoid response element (GRE) in the lipin gene. In fact, SMRT was recruited to the GRE, and this recruitment was dramatically up-regulated in the presence of dexamethasone. Thus, GR recruits SMRT in the presence of hormone, and SMRT then modulates glucocorticoid-mediated transcription. These data suggest that SMRT is a key regulator of metabolism in adipocytes, and that alterations in SMRT expression may dictate sensitivity to glucocorticoids in vivo.[br][br]Sources of Research Support: NIH R01 DK078125.[br][br]Nothing to Disclose: SM, XH, ML, RNC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1660 314 2405 MON-527 PO23-01 Monday 2074 2012


2070 ENDO12L_MON-528 POSTER SESSION: Nuclear Receptors: Coactivators [amp] Corepressors (1:30 PM-3:30 PM) Investigation of Estrogen Receptor [alpha] [mdash] Coregulator Complexes Bound to Natural Human Gene Promoter/Enhancer(s) Ross A Hamilton, Charles E Foulds, David M Lonard, Jun Qin, Bert W O[apos]Malley Baylor College of Medicine, Houston, TX The composition of coregulators (CoRs) recruited to estrogen receptor [alpha] (ER[alpha]) bound to its cognate DNA-binding site in a cell remains unknown, as chromatin immunoprecipitation (ChIP) assays have at best only revealed single ER[alpha]-CoR interactions. Only by the use of estrogen response element (ERE) DNA pulldown assays can detailed mechanistic proteomic information be ascertained.[br]To complement our ongoing analysis of ERE pulldown assays employing multiple copies of artificial canonical EREs, recombinant ER[alpha], and nuclear extracts (NEs) as a source of associated transcriptional CoRs, we have initiated studies to determine the CoR complexes formed on 300-400 base pair natural human genomic fragments containing EREs from two well-characterized ERa target genes- TFF1 (pS2) and GREB1. From testing candidate CoRs observed bound to artificial EREs by Western blotting, we found that the first ERE of the GREB1 proximal promoter (ERE1) bound a similar subset of CoRs with E2-liganded ER[alpha] as seen on artificial EREs. Unexpectedly, the first ERE of the pS2 proximal promoter (ERE1) only showed SRC/p160 family members recruited with E2-liganded ER[alpha]. Mass spectrometry of GREB1 and pS2 complexes revealed that distinct CoR components are bound to the different ERE-containing fragments. We have found on artificial EREs that kinase activities from NE are recruited to E2-liganded ER[alpha] complexes, resulting in phosphorylation of many, but not all, CoRs. Interestingly, the GREB1 ERE1, but not the pS2 ERE1, supports similar CoR complex phosphorylation. Mutagenesis of putative transcription factor binding sites, in addition to the ERE, should reveal how CoRs are recruited with ER[alpha] to the GREB1 fragment. Finally, we are assaying CoR composition on the most upstream ERE in the pS2 enhancer region (ERE3), and this data will be compared with the pS2 promoter proximal ERE1.[br][br]Sources of Research Support: This work was supported by grants HD08818 and NURSA from the National Institutes of Health.[br][br]Nothing to Disclose: RAH, CEF, DML, JQ, BWO 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2249 314 2406 MON-528 PO23-01 Monday 2075 2012


2071 ENDO12L_MON-529 POSTER SESSION: Nuclear Receptors: Coactivators [amp] Corepressors (1:30 PM-3:30 PM) Phosphorylation as a Potential Activation Mechanism for DNA-Bound Steroid Receptor-Coregulator Complexes Charles E Foulds, Suzanna L Bailey, Qin Feng, Ross A Hamilton, David M Lonard, Jun Qin, Bert W O[apos]Malley Baylor College of Medicine, Houston, TX The composition of coregulators (CoRs) recruited to a steroid receptor (SR) bound to its cognate DNA-binding site in a cell remains unknown, as chromatin immunoprecipitation (ChIP) assays can at best only detail a single SR-CoR interaction. Only by the use of hormone response element (HRE) DNA pulldown assays can detailed mechanistic proteomic information be ascertained.[br]We have designed HRE pulldown assays employing multiple copies of canonical estrogen or progesterone DNA response elements (EREs or PREs), recombinant SR (ERs or PRs), and nuclear extracts (NEs) as a source of associated transcriptional CoRs. We determined the CoR complexes formed on HREs- employing both naked and nucleosome-bound HREs.[br]Using both unbiased (mass spectrometry) and candidate-based (Western blotting) approaches, we found at least 16 CoRs with E2-liganded ERa on 4xEREs. Interestingly, both coactivators (SRCs) and corepressors (RIP140) are stably bound with E2-liganded ERa, whereas antagonists such as 4-HT recruit different corepressors (HDACs, SMRT, NCoR). Based on immunodepletion experiments, SRC-3 is the primary SRC/p160 family member for ERa-mediated [italic]in vitro[/italic] transcription, and TRIM24/TIF1a acts as a coactivator in our [italic]in vitro[/italic] transcription assays by being recruited to E2-liganded ERa only on nucleosomal templates (and not to PRs). Mechanistically, we found that HRE-SR-CoR complexes are stable after washes (resistant to high salt and urea) until fast phosphorylation events that appear to remove corepressors, converting [apos]poised[apos] complexes likely representing a gene [apos]OFF[apos] state, to ones [apos]activated[apos] for gene transcription. Furthermore, we observe phosphorylated residues of both SRC-3 and ERa published to be transcriptionally activating. We have identified the kinase that phosphorylates SRCs and ERa on EREs and have found that it plays a role in shifting CoRs complexes to the [apos]activated[apos] state.[br][br]Sources of Research Support: This work was supported by grants HD08818 and NURSA from the National Institutes of Health.[br][br]Nothing to Disclose: CEF, SLB, QF, RAH, DML, JQ, BWO 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1771 314 2407 MON-529 PO23-01 Monday 2076 2012


2072 ENDO12L_MON-530 POSTER SESSION: Nuclear Receptors: Coactivators [amp] Corepressors (1:30 PM-3:30 PM) Steroid Receptor Coregulator-2 Is Critical for Murine Mammary Morphogenesis and Human Breast Tumor Cell Proliferation Ramakrishna Kommagani, Maria M Szwarc, Francesco J DeMayo, John P Lydon Baylor College of Medicine, Houston, TX Steroid receptor coregulator-2 (SRC-2) is a member of the p160/SRC family of coregulators. Mouse studies demonstrate that these coregulators exert diverse regulatory effects [italic]in vivo[/italic]. Importantly, deregulation of these coregulators is recognized as a key causal factor in the etiopathogenesis of many mammalian target tissues. In the case of the mammary gland, SRC-1 and SRC-3 exert specialized roles not only in mammary morphogenesis but also in mammary tumorigenesis and metastasis. In contrast, the role of SRC-2 in mammary development and tumorigenesis has not been similarly investigated. To address this insufficiency, we used cre/loxP technology to generate a progesterone receptor (PR)[sup]cre/+[/sup]/SRC-2[sup]f/f[/sup] (SRC-2[sup]d/d[/sup]) bigenic mouse in which SRC-2 is selectively ablated in cells that express the progesterone receptor (PR). Absence of SRC-2 in PR positive cells in the mammary gland results in the inability of progesterone to induce mammary ductal side-branching and alveologenesis. Neither estrogen-driven epithelial ductal elongation at puberty nor prolactin-induced differentiation of the mammary epithelium in the adult requires SRC-2 function, highlighting a distinct coregulator role for SRC-2 in progesterone-dependent mammary proliferation. This conclusion is further supported by our recent molecular studies which show that SRC-2 is essential for the induction a wide-spectrum of molecular targets which mediate progesterone-initiated mitogenesis in the mammary epithelium. Significantly, one of these targets is receptor activator of nuclear factor kappaB ligand (RANKL), a cytokine that is essential for progesterone-dependent expansion of the mammary stem cell population during normal mammary morphogenesis and for progestin-promotion of mammary tumorigenesis. Moreover, expression studies reveal that SRC-2 levels are markedly elevated along with SRC-3 in both human and murine mammary tumor tissue, suggesting that the coregulator properties of SRC-2 may extend to mammary tumor promotion. To test this assertion, cell culture studies show that knockdown of SRC-2 using siRNA methods results in a marked decrease in the proliferation of T47D breast cancer cells in response to the progestin R5020. Together, these studies demonstrate that SRC-2 is required for progesterone-dependent mammary morphogenesis and that tight control of SRC-2 levels are essential to avoid aberrant proliferative responses that contribute to mammary tumor progression in both human and mouse.[br][br]Sources of Research Support: This research is supported by National Institutes of Health grants (U54 HD-07495) and (CA-077530).[br][br]Nothing to Disclose: RK, MMS, FJD, JPL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1133 314 2408 MON-530 PO23-01 Monday 2077 2012


2073 ENDO12L_MON-531 POSTER SESSION: Nuclear Receptors: Coactivators [amp] Corepressors (1:30 PM-3:30 PM) Steroid Receptor Coactivator-1 (SRC-1) and SRC-2 from Mouse Brain Physically Interact with Mouse Progestin Receptor Wing Yee Wan, DaEun Im, Kelsey L Diederich, Didem Vardar-Ulu, Jennifer K Hood-DeGrenier, Marc J Tetel Wellesley College, Wellesley, MA; Wellesley College, Wellesley, MA; Wellesley College, Wellesley, MA The steroid hormone, progesterone, acts in brain to influence a variety of events, including development and reproduction. Nuclear receptor coactivators are necessary for full ligand-dependent transcriptional activity of progestin receptors (PR) in vitro. Studies from our lab and others reveal that members of the p160 family of nuclear receptor coactivators, including steroid receptor coactivator-1 (SRC-1) and SRC-2, act in the hypothalamus to modulate PR-dependent reproductive behavior in female rodents. In the present studies, we used pull-down assays to test the hypothesis that SRC-1 and SRC-2 from female mouse hypothalamic whole cell extracts physically associate with mouse PR in a ligand-dependent manner. Using GST-tagged mouse PR-A and PR-B expressed in baculovirus cells, we found that SRC-1 from hypothalamic tissue physically interacted more with PR-A in the presence of agonist R5020 (mean ratio of hypothalamic SRC-1/1% input[plusmn]SEM, 1.4[plusmn]0.1) than in the absence of ligand (0.6[plusmn]0.1) or in the presence of the selective PR modulator, RU486 (0.6[plusmn]0.1; p[lt]0.001). Similarly, SRC-2 from hypothalamus associated more with PR-A when bound to agonist (1.3[plusmn]0.1) than in the absence of ligand (0.4[plusmn]0.1) or when bound to RU486 (0.3[plusmn]0.1; p[lt]0.001). In contrast to mouse PR-A, neither SRC-1 nor SRC-2 from hypothalamus interacted with mouse PR-B in a ligand-dependent manner. These findings indicate that SRC-1 and SRC-2 from mouse hypothalamus interact with PR-A, but not PR-B, in a ligand-dependent manner. The results from these studies, using biologically-relevant brain tissue, suggest that SRC-1 and SRC-2 from the hypothalamus interact with mouse PR in an isoform-specific manner. Further investigation of these interactions between coactivators from brain and PR will provide insights into the differential mechanisms of PR isoforms in brain and may allow for the identification of novel cofactors involved in the steroid receptor complex.[br][br]Sources of Research Support: This research was supported by NIH Grant R01 DK61935 awarded to MJT.[br][br]Nothing to Disclose: WYW, DI, KLD, DV-U, JKH-D, MJT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1248 314 2409 MON-531 PO23-01 Monday 2078 2012


2074 ENDO12L_MON-532 POSTER SESSION: Nuclear Receptors: Coactivators [amp] Corepressors (1:30 PM-3:30 PM) Genomic Recruitment of the p160 Family of Steroid Receptor Coactivators (SRCs) in Prostate Cancer Cells Bin He, Rainer Lanz, Chuandong Geng, Sue Anne Chew, Vijay Kumar Eedunuri, Bert W O[apos]Malley, Nicholas Mitsiades Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Adrienne Helis Malvin Medical Research Foundation, New Orleans, LA [bold]Background:[/bold] In castration-resistant prostate cancer (CRPC), androgen receptor (AR) signaling persists despite low circulating testosterone levels. Overexpression of the p160 Steroid Receptor Coactivators (SRCs), that mediate AR transcriptional activity, is one of the mechanisms contributing to CRPC recurrence. Recently, SRC2 gene amplification was found in a subset of human prostate carcinomas (PC), suggesting a role as an oncogene. It is critical to define the precise function of each SRC and their respective contribution to AR transcriptional activity in PC cells. However, there is no comprehensive unbiased genome-wide analysis of SRC chromatin binding in PC. In this study, using the combination of massive parallel sequencing platforms with chromatin immunoprecipitation (ChIP-Seq), we investigated the role of each SRC in the transcriptional regulation of PC cells.[br][bold]Methods:[/bold] LNCaP cells cultured in RPMI1640 medium supplemented with 10% FBS (which leads to [sim]80% of maximal AR activity, based on AR target gene expression) were fixed in freshly-prepared 1% formaldehyde. ChIP, Illumina ChIP-Seq library construction, DNA sequencing, MACS peak calling, and bioinformatic analysis were performed.[br][bold]Results:[/bold] We identified 39,845 AR, 13,567 SRC1, 25,948 SRC2, and 4,214 SRC3 binding sites. Among SRCs, SRC2 has the highest number of binding sites (FDR 0.45%), while SRC3 has the lowest number, even though the MACS cutoff for SRC3 was less stringent (FDR 11.53%). Interestingly, 94.4%, 87.8% and 89.4% of the SRC3 binding sites overlap with those of AR, SRC1 and SRC2, respectively. AR and SRC binding to select sites was confirmed by ChIP-PCR. Nevertheless, only about half of AR binding sites overlap with any SRC binding site. We are currently exploring the significance of the SRC-independent AR binding sites by integrating these results with gene expression datasets from LNCaP cells treated with AR siRNA.[br][bold]Conclusions:[/bold] The p160 SRCs play important roles in AR transcriptional activity in PC. There is significant overlap between AR, SRC1, SRC2 and SRC3 genomic binding events. Among the three p160 SRCs, SRC2 is recruited to the largest number of binding sites in LNCaP cells, while SRC3 is recruited to a significantly smaller number of binding sites. We propose that SRC2 plays a major role in PC and could be an effective therapeutic target for treatment of CRPC.[br][br]Sources of Research Support: The authors acknowledge the joint participation by Adrienne Helis Malvin Medical Research Foundation through its direct engagement in the continuous active conduct of medical research in conjunction with Baylor College of Medicine. B.H. is supported by NIH/NIDDK grant 5K01DK081446. N.M. is a Dan L. Duncan Scholar and a Caroline Wiess Law Scholar at Baylor College of Medicine and his work is also supported by the Conquer Cancer Foundation of the American Society of Clinical Oncology (ASCO) Career Development Award and by the Prostate Cancer Foundation.[br][br]Nothing to Disclose: BH, RL, CG, SAC, VKE, BWO, NM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1874 314 2410 MON-532 PO23-01 Monday 2079 2012


2075 ENDO12L_MON-533 POSTER SESSION: Nuclear Receptors: Coactivators [amp] Corepressors (1:30 PM-3:30 PM) Small Molecule Protein Kinase C Inhibitors as Promising Therapeutic Agents for Prostate Cancer Treatment Vijay Kumar Eedunuri, Diego Jacinto Bedoya, Sue Anne Chew, Ashesh Shah, Bin He, David Lonard, Bert W O[apos]Malley, Nicholas Mitsiades Adrienne Helis Malvin Medical Research Foundation, New Orleans, LA; Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX [bold]Background:[/bold] The Steroid Receptor Coactivators (SRCs) of the p160 family are key pleiotropic [ldquo]master regulators[rdquo] of transcription factors necessary for cancer cell proliferation, survival, metabolism and metastasis. The androgen receptor (AR) plays crucial roles in prostate cancer (PC) growth and disease progression. Depletion of SRCs can suppress AR signaling and cellular proliferation of PC cells, suggesting that the SRCs could be therapeutic targets in PC. We have previously reported that protein kinase C (PKC) family members can phosphorylate SRC-3 (AIB1/NCoA3) and prevent its proteasome-mediated degradation. In this study, we explored the impact on SRC expression and AR function of three PKC small molecule inhibitors (SMIs): Bisindolylmaleimide I (BIM), midostaurin (PKC412) and Sangivamycin.[br][bold]Methods:[/bold] MTT assay was used to quantify cell viability of PC cell lines after treatment with PKC SMIs and the combination of these SMIs with other existing anticancer agents. To determine the impact of the PKC SMIs on SRC-3 protein and mRNA levels, we used immunoblotting and RT-qPCR, respectively.[br][bold]Results:[/bold] Sangivamycin, BIM and PKC412 successfully decreased the growth and cell viability of LNCaP, 22Rv1 and LAPC4 cells. Treatment with these PKC SMIs depleted SRC-3 protein, leading to disruption of AR signaling, as reflected by decreased expression of the AR target genes KLK3 (PSA), TMPRSS2 and FKBP5, and enhanced the anticancer activity of the antiandrogen MDV3100. Sangivamycin treatment resulted in increased Akt phosphorylation. Combination of this SMI with the Akt inhibitor MK2206 resulted in greater than additive activity against 22Rv1 cells.[br][bold]Conclusions:[/bold] PKC SMIs can post-translationally destabilize SRC-3 protein, disrupting AR signaling and decreasing PC cell growth and viability. Thus, PKC inhibitors are promising agents for the treatment of PC.[br][br]Sources of Research Support: The authors acknowledge the joint participation by Adrienne Helis Malvin Medical Research Foundation through its direct engagement in the continuous active conduct of medical research in conjunction with Baylor College of Medicine. B.H. is supported by NIH/NIDDK grant 5K01DK081446. N.M. is a Dan L. Duncan Scholar and a Caroline Wiess Law Scholar at Baylor College of Medicine and his work is also supported by the Conquer Cancer Foundation of the American Society of Clinical Oncology (ASCO) Career Development Award and by the Prostate Cancer Foundation.[br][br]Nothing to Disclose: VKE, DJB, SAC, AS, BH, DL, BWO, NM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1876 314 2411 MON-533 PO23-01 Monday 2080 2012


2076 ENDO12L_MON-534 POSTER SESSION: Nuclear Receptors: Coactivators [amp] Corepressors (1:30 PM-3:30 PM) Roles of Tyrosine Residues in Multiple Functions of p54nrb Liangliang Liu, Wayne Hung, Xuesen Dong Vancouver Prostate Centre, Vancouver, Canada The RNA binding protein p54nrb exerts multiple functions and participates in a variety of nuclear processes, including transcription initiation, RNA processing, and DNA repair. p54 binds steroid receptors and further recruits the Sin3A/HDAC transcription repressor complex to inhibit downstream gene transcription initiation. p54 also forms a protein complex with the spliceosome to play essential roles in pre-mRNA splicing. These multiple functions of p54 could regulate gene transcription both positively and negatively, but the molecular mechanism is unclear.[br]Our previous work demonstrates that protein phosphatase-1 dephosphorylates p54 at serine and threonine residues, alters p54 protein associations and therefore modulates p54 transcription corepression and RNA splicing activities. In this report, we further demonstrate that the tyrosine residue 267 is also important for the many functions of p54. Using luciferase reporter assays, we show that p54wt induces 50% decreases in both the androgen receptor (AR) and the progesterone receptor (PR) transactivation. Mutation of tyrosine 267 into phenylalanine, p54(Y267F), results in 2 and 6 fold increases in PR and AR transactivation, respectively. In our RNA splicing assays, we show that p54wt induces 30-40% increases of both ARwt and ARv7 (an AR splice variant) transcripts, while p54(Y267F) results in a 70% decrease of ARv7 transcripts. In addition, we show that p54(Y267F) decreases protein interactions with the HDAC1 corepressor, and U1A and U2AF RNA splicing factors when compared to p54wt.[br]We conclude from these data that tyrosine residue 267 plays important roles in regulating the multiple functions of p54.[br][br]Nothing to Disclose: LL, WH, XD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2161 314 2412 MON-534 PO23-01 Monday 2081 2012


2077 ENDO12L_MON-535 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) The Nuclear Receptor DNA Binding Project: A New Web Resource Frances M Sladek, Bin Fang, Daniel Mane-Padros, Eugene Bolotin, Tao Jiang, Thomas Girke University of California, Riverside, CA; University of California, Riverside, CA; University of California, Riverside, CA; University of California, Riverside, CA; Children[apos]s Hospital Oakland Research Institute, Oakland, CA Despite 25 years of intense investigation of nuclear receptors (NRs), much remains to be learned about exactly what DNA sequences these transcription factors bind and where. The post-genomic era of biomedical research also necessitates a more global approach to this important issue. Here we present the Nuclear Receptor DNA Binding Project that establishes an interactive web portal to comprehensive data on NR binding motifs, direct target genes and single nucleotide polymorphisms (SNPs) in NR binding sites. Primary data are generated using a high throughput assay termed protein binding microarrays (PBMs). PBMs contain up to a million spots of double-stranded DNA to which full length nuclear receptors in a physiologically relevant environment are added (crude nuclear extracts from mammalian cells, in the presence or absence of ligand). This high throughput DNA binding assay allows for the generation of large amounts of data on NR DNA binding specificity, as well as affinity, that can be used to help predict new target genes and distinguish overlapping DNA binding specificity among NRs. The PBM data are cross referenced with publicly available genome-wide location studies (ChIP-seq) as well as expression profiling studies (e.g., Transcriptomine). The PBM technology is also used to identify SNPs in NR response elements that alter the binding of the NR to DNA (affinity altering SNPs or aaSNPs). aaSNPs are coupled with data from Genome Wide Association Studies (GWAS) and the Genotype-Tissue Expression (GTEx) database to predict altered susceptibility to disease. Identification of aaSNPs in NR binding sites will help reveal the relevance of certain genetic differences between individuals and may help elucidate health disparities. We establish the pipeline with the first set of NRs -- HNF4, RXR, COUPTF and the PPARs -- and present the searchable web-based tools and datasets associated with the PBM data. The Nuclear Receptor Binding Project is linked to the Nuclear Receptor Signaling Atlas (NURSA, www.nursa.org), the Cistrome Project (www.cistrome.org) and the Transcription Factor Encyclopedia (Tfe, http://www.cisreg.ca/cgi-bin/tfe/home.pl). The goal is ultimately to send all 48 human NRs (with a variety of ligands) through the pipeline and establish a publicly available on-line resource that will help fast-track identification of direct NR target genes and pave the way for personalized medicine.[br][br]Sources of Research Support: This work is funded by NIH grants to FMS and TJ (R21MH087397 and 1R01DK094707).[br][br]Nothing to Disclose: FMS, BF, DM-P, EB, TJ, TG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2130 315 2413 MON-535 PO23-02 Monday 2082 2012


2078 ENDO12L_MON-536 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) Cistromes of Human CAR Reveal Novel Regulation Mechanisms Rui Xiao, Stephen Ayers, David D Moore Baylor College of Medicine, Houston, TX; Methodist Hospital Research Institute, Houston, TX Constitutive androstane receptor (CAR) belongs to the NR1I subgroup of nuclear receptor family and is closely related to other two members of the subgroup vitamin D receptor (VDR) and Pregnane X receptor (PXR). CAR and PXR are considered as [ldquo]xenobiotic sensors[rdquo], but are also activated by endobiotics. Transcriptional activity of CAR can be activated by agonist ligands or non-ligand activator, such as phenobarbital. Studies have shown that CAR plays important roles in xenobiotic/endobiotic metabolism, tumorigenesis, glucose/lipid metabolism, etc. Like many nuclear receptors, CAR directly binds to DNA to regulate target gene expression by forming heterodimers with RXR[alpha]. We used an in vivo biotinylation system to determine the cistromes of human CAR (hCAR) in the presence of various ligands in HepG2 cells. Stable clones that express biotin ligase BirA and hCAR were established. Expression of endogenous known CAR target genes was strongly induced by agonists and suppressed by inverse agonists. Biotin-mediated ChIP assay (biotin-ChIP) showed very robust CAR binding at known CAR response elements. Genome-wide biotin-ChIP with next generation sequencing indicates that different ligands change the cistromes of CAR, and that inverse agonists do not generally reduce DNA binding of CAR. Pathway analysis of target genes identified with CAR cistromes and microarray expression data reveals both known and potential novel functions of CAR. Motif analysis shows that CAR predominantly binds to direct repeat 4 motifs, and that several known transcription factor motifs are enriched in CAR binding regions including HNF4, CEBPA, AP1, FOXA, etc. Surprisingly, cistromes of CAR significantly overlap with those of RXRA/HNF4A/CEBPB/JUND determined in HepG2 cells that do not express detectable hCAR. ChIP/qPCR confirms that CAR either binds to regions that are preloaded with these transcription factors, or induces [italic]de novo[/italic] and synergistic recruitments of these TFs at others regions in response to ligand treatment. Our results suggest that CAR coordinates with other transcription factors in gene transcription regulation in gene-specific manners.[br][br]Nothing to Disclose: RX, SA, DDM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1883 315 2414 MON-536 PO23-02 Monday 2083 2012


2079 ENDO12L_MON-537 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) Nuclear Receptor Constitutive Androstane Receptor CAR as a Target for Cancer Therapy Nan Wu, Bingning Dong, David D Moore Baylor College of Medicine, Houston, TX Constitutive androstane receptor CAR (NR1I3) is a nuclear receptor with constitutive transactivation function (1) and is highly expressed in the liver and the small intestine. CAR is essential in a variety of physiological functions, such as metabolism of xenobiotics, bilirubin, and bile acids (2, 3). Short-term CAR activation promotes liver growth and DNA synthesis (2) but long-term activation of CAR results in tumorigenesis. To understand the role of CAR in viral liver tumorigenesis, we used the mice with transgenic expression of the hepatitis B virus x antigen (ATX mice) to determine CAR promotion of hepatocellular carcinoma (HCC) (4, 5). We injected mutagen diethylnitrosamine (DEN) 25mg/kg to 10-12 day old male ATX mice, and sacrificed mice at 2 month and 4 month. At 4 month, we found DEN-treated ATX mice had developed tumor nodules and mRNA expression of the CAR target gene (CYP2B10) increased to 3.5 folds in ATX mice treated with DEN compared with ATX mice treated with PBS. It demonstrates CAR activity is induced during tumorigenesis. To characterize the molecular mechanism of CAR function in tumorigenesis of ATX mice, we searched for CAR targets of cell cycle genes. ICR mice (wild type mice of ATX mice) were treated with a single dose of TCPOBOP (3 mg/kg, i.p.) and six injections of 3-hydroxy-5[plusmn]androstanol (100 mg/kg, Steraloids, Newport, RI), dissolved in corn oil, 72, 48, 24 and 1 h before TCPOBOP (a mouse CAR agonist) and 24, 48 h afterwards. TCPOBOP alone, androstanol alone or oil, was given to three additional groups of mice respectively. Mice were sacrificed 72 hours after TCPOBOP treatment. We found androstanol inhibited TCPOBOP-induced drug metabolic genes (CYP2B10, CYP3A11 and CYP2C29). Besides drug metabolic genes, FoxM1 mRNA was induced to 9 folds by TCPOBOP and then was inhibited to 35% of mRNA expression in TCPOBOP group by androstanol treatment. FoxM1 stimulates proliferation by promoting S-phase and M-phase entry and contributes to both tumor initiation and progression. This suggests that CAR plays a positive role in DEN-initiated tumorigenesis and inhibition of CAR activation by androstanol is a potential target for cancer therapy.[br][br](1) Baes M et al., Mol Cell Biol 1994; 14:1544. (2) Wei P et al., Nature 2000; 407(6806):920. (3) Huang W et al., J Clin Invest 2004; 113:137. (4) Tsai WL et al., Oncogene 2010; 29(16):2309. (5) Feitelson MA et al., Cancer Lett 2009; 286(1):69.[br][br]Nothing to Disclose: NW, BD, DDM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1802 315 2415 MON-537 PO23-02 Monday 2084 2012


2080 ENDO12L_MON-538 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) Profiling of Nuclear Receptor Expression in Breast Cancer[ndash]Associated Fibroblasts Identifies RORA, THRB, VDR and PPARG as Potential Mediators within the Tumor Microenvironment Kevin C Knower, Ashwini Chand, Natalie Eriksson, Kiyoshi Takagi, Yasuhiro Miki, Hironobu Sasano, Geoff Lindemann, Jane Visvader, George Muscat, Evan R Simpson, Colin D Clyne Prince Henry[apos]s Institute, Melbourne, Australia; University of Queensland, Brisbane, Australia; Tohoku University Graduate School of Medicine, Sendai, Japan; The Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia The interaction between breast tumour epithelial and stromal cells is vital for initial and recurrent tumour growth. The cancer-associated stroma is reprogrammed to provide the most opportune environment for tumour proliferation and metastasis. The extent of this reprogramming is poorly understood. Nuclear receptors (NRs) are intracellular transcription factors that directly regulate gene expression in response to lipophilic molecules. In the cellular context NRs regulate cell proliferation, differentiation and apoptosis mainly via the transcriptional regulation of target genes and as major points of convergence of multiple signal transduction pathways. NRs such as Estrogen Receptor-[alpha] (ER[alpha]) and Progestin Receptor (PR) are the most important indicators of breast tumour type and stage, response to treatment and prognosis. However, little is known about the status of these and the complete family of NRs in tumour associated stroma. In the current study, all forty-eight NRs in normal breast adipose fibroblasts (BAFs) and breast cancer-associate fibroblasts (CAFs) were screened for differing expression profiles.[br]Nuclear Receptor Low Density Taqman Arrays were used to compare the gene expression profiles in a collection of primary cultured CAFs (n=9) and BAFs (n=7). StatMiner Analytical software was used to perform Non-Parametric (Wilcoxon) tests while geNorm selected the most stable endogenous controls used for normalisation. Initial analyses revealed that thirty-three of 48 NRs were expressed in the stromal cells distributed over the following groups - Endocrine Receptors (11/12), Adopted Orphan Receptors (8/11) and Orphan Receptors (14/25). Of the NRs expressed, four showed significant down-regulation in CAFs compared to BAFs [ndash] RAR-related orphan receptor-[alpha] (RORA) (1.95-fold); Thyroid hormone receptor-[beta] (THRB) (15.63-fold); Vitamin D receptor (VDR) (1.91-fold); and Peroxisome proliferator-activated receptor-[gamma] (PPARG) (3.48-fold). No NRs were found to be significantly upregulated in the CAFs compared to BAFs.[br]Silencing of NRs in the stromal microenvironment may prove an important component of the cross-talk between neighbouring epithelial cells driving tumour growth. The identification of the molecular mechanisms and functional consequences of the silencing of these four NRs in CAFs is currently being determined and may provide an avenue for the development of intratumoural-targeted therapies.[br][br]Nothing to Disclose: KCK, AC, NE, KT, YM, HS, GL, JV, GM, ERS, CDC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1512 315 2416 MON-538 PO23-02 Monday 2085 2012


2081 ENDO12L_MON-539 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) The Nuclear Receptor Ror[beta] Modulates Expression of Genes Involved in Extracellular Matrix Regulation and Runx2 Activity Matthew M Roforth, Gang Liu, Kumar Chokalingam, Sundeep Khosla, David G Monroe Mayo Clinic College of Medicine, Rochester, MN The regulation of osteoblastic differentiation requires the coordinate activities of a number of transcription factors that ultimately direct the production of a mineralization-competent extracellular matrix (ECM). In a previous screen designed to assess the expression patterns of the nuclear receptor superfamily throughout osteoblastic differentiation, we identified retinoic acid receptor-related orphan receptor beta (Ror[beta]) as potently suppressed throughout differentiation. Although past reports state that Ror[beta] expression is spatially limited to the brain and retina, robust Ror[beta] expression was detected in whole bone, primary bone marrow stromal cells, calvarial cells, and osteocytes, but not in osteoclast cultures, suggesting Ror[beta] expression is limited to the mesenchymal osteoblastic lineage. Moreover, Ror[beta] overexpression inhibited mineralization of MC3T3-E1 cells. These data clearly indicate that Ror[beta] plays an influential role in osteoblast differentiation, however the molecular targets of Ror[beta] have not been identified in bone. In order to decipher the possible role of Ror[beta] in osteoblast differentiation, we employed whole transcriptome profiling followed by [italic]in silico [/italic]analysis using a MC3T3 cell model where Ror[beta] is modestly overexpressed. We identified 317 Ror[beta]-regulated genes ([ge] 1.5-fold; Q-value [le] 0.05) when compared to control MC3T3 cells. Metacore pathway analysis indicated that Ror[beta] regulates genes involved in ECM production and maintenance. Specifically, upregulation of genes involved in epithelial-to-mesenchymal transition (Notch1, Fn1, Tgfb2, Ednra) and ECM remodeling (Egfr, Serpine2, Dcn, Itga10, Itgb5) was observed. Consistent with the anti-osteogenic role of Ror[beta], stimulation of factors involved in ECM breakdown (Mmp12, Sdc2, Mgp) and suppression of the Mmp inhibitor Timp1 was observed. Previous data demonstrated that Ror[beta] may also suppress bone formation through inhibition of Runx2 activity, the major regulator of osteoblast differentiation and function. Interestingly Ror[beta] upregulated Zfp521, a known inhibitor of Runx2 signaling. Collectively, these data suggest that Ror[beta] may suppress osteoblast differentiation through two distinct mechanisms; 1) the disruption of ECM components needed for mineralization and 2) suppression of Runx2 activity by itself and/or through stimulation of Zfp521. Further characterization of Ror[beta] function in osteoblasts may define it as a potential clinical target to treat age-related bone loss.[br][br]Sources of Research Support: NIH Grant AG004875; Mayo Kogod Center for Aging.[br][br]Nothing to Disclose: MMR, GL, KC, SK, DGM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1553 315 2417 MON-539 PO23-02 Monday 2086 2012


2082 ENDO12L_MON-540 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) Hypoxia-Dependent Regulation of [beta]-Catenin by the Orphan Nuclear Receptor Nur77 Sally KY To, Jin Z Zeng, Alice ST Wong University of Hong Kong, Hong Kong, China; Xiamen University, Xiamen , China It has become increasingly recognized the importance of tumor microenvironment in cancer initiation and progression. The microenvironment of solid tumors contains regions of poor oxygenation as a result of hypoxia. Nur77, an orphan nuclear receptor, can play multiple roles in cell survival and apoptosis in a cell-context dependent manner. Nur77 is activated in colon cancers and in the large majority of cases by an increased or altered expression. Since Nur77 gene amplification is a rare event, the mechanisms underlying Nur77 protein expression in colon tumors remain obscure. Here we showed that hypoxia activated the expression of Nur77, resulting in higher levels of Nur77 protein and mRNA. We also showed for the first time that Nur77 overexpression resulted in an upregulation of [beta]-catenin expression under hypoxia. Knockdown of Nur77 using small interfering RNA significantly inhibited the activation of [beta]-catenin, confirming that the effect was Nur77 specific. Using paired colon carcinoma cell lines and various mutant constructs, we provided evidence that the Nur77-induced [beta]-catenin expression at the levels achieved by hypoxia was independent of both p53 and APC, which are components of two major pathways for [beta]-catenin degradation. Moreover, Nur77 lacking the DNA binding domain failed to regulate the expression of [beta]-catenin, suggesting that nuclear Nur77 is important for this regulation. These results may provide a molecular basis for overexpression of Nur77 in colon cancer and a novel regulatory mechanism that regulates cell survival and death.[br][br]Sources of Research Support: NSFC/RGC grant N_HKU 735/09.[br][br]Nothing to Disclose: SKYT, JZZ, ASTW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1557 315 2418 MON-540 PO23-02 Monday 2087 2012


2083 ENDO12L_MON-541 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) DY131 Regulation of the Akt-mdm2-p53 axis Mediated by Human Estrogen-Related Receptor [beta] in Prostate Cancer Cells Yuan Lu, Nicholas JE Starkey, Sara K Drenkhahn, Glenn A Jackson, Dennis B Lubahn University of Missouri, Columbia, MO; University of Missouri, Columbia, MO; University of Missouri, Columbia, MO; University of Missouri, Columbia, MO Several nuclear receptors, including the Estrogen Receptors and Androgen Receptor, have been shown to regulate the tumor suppressor p53. Here we investigate the ability of human Short-Form Estrogen Related Receptor beta (SFhERR[beta]) to interact with p53/p21 through Akt-mdm2.[br]We hypothesize that growth inhibitory effect of DY131, an ERR[beta] agonist, on prostate cancer (PCa) cells is due to the regulation of p53/p21 signaling pathway through the interaction between ERR[beta] and Akt-mdm2 axis. To test this we transfected SFhERR[beta] into the human PCa cell line DU145 and treated with the ERR[beta] agonist DY131. We monitored the growth inhibitory effect of DY131 on PCa cell lines by the treatment of DY131. To determine the pathway activity, we monitored the protein concentration of p53, p53-pSer392, p21/WAF1, mdm2, mdm2-pSer166, Akt and Akt-pSer473 via western blot analysis.[br]Results: We found that DY131 is able to increase p53 phosphorylation at Ser392. We also found that the p53 target gene, cyclin-dependent kinase inhibitor p21/WAF1, is upregulated by DY131 treatment; p53 negative regulator mdm2 is down-regulated by DY131 treatment. Akt activity, as indicated by Ser473 phosphorylation, can be inhibited by 1uM, 10uM and 20uM DY131, and overexpression of SFhERR[beta] amplified this effect. In addition, DY131 inhibited growth in both mouse and human PCa cell lines (TRAMPC2, IC50[sim]20uM; DU145, IC50 [sim]20uM). Our data indicates that DY131 can stimulate the p53 signaling pathway by inhibiting the activity of Akt-mdm2 axis function.[br]Conclusion: Our results suggest a new level of regulation of SFhERR[beta] on the p53 and p21/WAF1 signaling pathway, induced by DY131, which involves Akt[apos]s regulation on mdm2. Additionally, our results indicate that modulation of ERR[beta], via ERR[beta] selective drugs like DY131, may serve as a new target for PCa treatment.[br][br]Nothing to Disclose: YL, NJES, SKD, GAJ, DBL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1987 315 2419 MON-541 PO23-02 Monday 2088 2012


2084 ENDO12L_MON-542 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) Dax-1 Regulation of the Wnt Signaling Pathway in Mouse Embryonic Stem Cells Anthony Torres, Christina Tzagarakis-Foster University of San Francisco, San Francisco, CA The orphan receptor Dax-1 is highly expressed in pluripotent embryonic stem (ES) cells and shows a correlative reduction in expression as these cells differentiate. While it is known that Dax-1 is expressed in pluripotent mouse ES cells, the precise function of Dax-1 in these cells is not as well understood. Recent studies employing RNA interference (RNAi) to specifically reduce the expression of the Dax-1 gene in mouse ES cells found that upon the knock down of Dax-1, ES cells differentiated. These findings indicate that Dax-1 functions in a novel role in the maintenance of a relatively undifferentiated state in ES cells.[br]Dax-1 is important in early embryonic development and, when mutated, adrenal insufficiency and disruption of normal tissue architecture results. In our study, we silenced Dax-1 expression in mouse ES cells using RNAi followed by PCR array methodology to identify genes that were alternately regulated upon targeted knock-down of Dax-1. Several novel Dax-1 targets have been identified, including genes that are involved in the Wnt signaling pathway. In attempts to understand the mechanism of Dax-1 action in ES cells, we are investigating the effect of Dax-1 mutations on the regulation and expression of the identified target genes. A correlation to patient-relevant DAX-1 mutations seen in Adrenal Hypoplasia Congenita (AHC), a disease that is caused by mutations in the DAX-1 gene in humans, may be determined.[br][br]Sources of Research Support: This research is supported by the University of San Francisco, College of Arts and Sciences, Faculty Development Fund.[br][br]Nothing to Disclose: AT, CT-F 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2282 315 2420 MON-542 PO23-02 Monday 2089 2012


2085 ENDO12L_MON-543 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) Nuclear Receptor NR5A1 Prevents Centrosome Over-Duplication by Restricting Aberrant Activation of DNA-Dependent Protein Kinase in the Centrosome Chia-Yih Wang, Bon-Chu Chung Academia Sinica, Taipei, Taiwan Steroidogenic factor 1 (SF-1, NR5A1, Ad4bp) is a nuclear receptor that controls adrenogonadal cell growth and differentiation. Here we show that SF-1 restricts centrosome over-duplication by inhibiting aberrant activation of DNA dependent protein kinase (DNA-PK). SF-1 resides in the centrosome, where it interacted with Ku70/Ku80 and sequestered them from their catalytic subunit DNA-PKcs. In the absence of SF-1, activated DNA-PKcs was recruited to the centrosome, leading to aberrant phosphorylation of Akt and accumulation of Cyclin A/CDK2 in the centrosome but not in the whole cell. Consistent with the lack of global DNA-PK activation, DNA damage response was not detected. Centriole biogenesis and cell cycle progression were also not affected by SF-1 depletion. The centrosome over-duplication defect caused by SF-1 depletion was averted by the elimination of DNA-PKcs, Cyclin A, or CDK2, or Akt inhibition. Thus, SF-1 prevents centrosome over-duplication by putting centrosomal DNA-PK and CDK2 signaling into check without affecting DNA damage response in the nucleus.[br][br]Nothing to Disclose: C-YW, B-CC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1404 315 2421 MON-543 PO23-02 Monday 2090 2012


2086 ENDO12L_MON-544 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) Altered Expression of Metabolic Nuclear Receptor Target Genes in Atp7b[sup]-/-[/sup] Mice (Wilson Disease Mouse) Clavia Ruth Wooton-Kee, Michael A Grusak, Svetlana Lutsenko, David D Moore Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Johns Hopkins University, Baltimore, MD Wilson[apos]s disease is an autosomal recessive disease due to mutations in the Cu-transporting P-type ATPase (ATP7b) and results in excessive hepatic copper accumulation and often results in liver failure. The Atp7b[sup]-/-[/sup] mouse animal model has increased hepatic copper levels similar to those of Wilson[apos]s disease patients as well as increased nuclear copper concentrations. Previous studies demonstrated that copper has a high affinity for the estrogen receptor DNA binding domain (ER-DBD), and incubation of the ER-DBD with copper results in loss of ER binding to response elements. Our hypothesis is that elevated hepatic copper concentrations result in decreased nuclear receptor activation due to disruption of zinc binding in the DNA-DBD of nuclear receptors. [italic]In vitro[/italic] translation of RXR with 4-40 [mu]M of copper (CuSO4) resulted in complete loss of FXR:RXR binding to the short heterodimer partner protein (SHP) and bile salt export pump (Bsep, canalicular bile acid transporter) promoters, and the addition of 40 [mu]M zinc (ZnSO4) to the reaction restored binding to the SHP and BSEP promoters. Treatment of HepG2 cells with 5 mM CuSO4 decreased chenodeoxycholic acid mediated activation of BSEP mRNA expression. In Atp7b[sup]-/-[/sup] mice at 6, 12, and 16-20 weeks Bsep and SHP mRNA expression were decreased 40% and 40-20% along with decreased binding of FXR to the Bsep and SHP mouse promoters in EMSA analysis. The mRNA expression of other nuclear receptor target genes were also decreased in the Atp7b[sup]-/-[/sup] mice at 6, 12, and 16-20 weeks: HNF4a (Na[sup]+[/sup]-taurocholate transporter protein, basolateral bile salt transporter, decreased 20%), LRH-1 (Cyp8b, required for cholic acid synthesis, decreased 40%; Abcg8, canalicular cholesterol transporter, decreased 20%), and TR (Spot14, regulation of triglyceride pools, decreased 40 [ndash] 20 %). The mRNA expression of HNF4a, and protein expression of RXR were unchanged in the Atp7b[sup]-/-[/sup] mice. The basal mRNA expression of phase-I enzymes Cyp3a11 and Cyp2b10 were increased 2-fold (P= 0.057) in the Atp7b[sup]-/-[/sup] mice, but activation with the constitutive androstane receptor (CAR) agonist TCPOBOP was attenuated in the knockout mice (20%, Cyp3a11 and 10% Cyp2b10) relative to wild-type littermates. These data show that the Wilson[apos]s disease mouse model has defects in nuclear receptor signaling that impair the regulation of genes involved in maintaining metabolic homeostasis in the liver.[br][br]Sources of Research Support: T32DK007664-19; F32DK089689-01A1.[br][br]Nothing to Disclose: CRW-K, MAG, SL, DDM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2060 315 2422 MON-544 PO23-02 Monday 2091 2012


2087 ENDO12L_MON-545 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) Transcriptional Regulation of Autophagy by Nutrient-Sensing Nuclear Receptors Jae Man Lee, Martin Wagner, Rui Xiao, David D Moore Baylor College of Medicine, Houston, TX Autophagy is an evolutionally conserved catabolic process involving the degradation of cytoplasmic components through the lysosomal machinery. When properly controlled, autophagy contributes to homeostatic responses to internal and external stimuli, whereas the abnormal activity of autophagy is associated with a broad spectrum of human pathophysiology and disease. These include development, aging, innate and adaptive immunity, muscle diseases, Crohn[apos]s disease, cancer, neurodegenerative diseases, diabetes, and metabolic disorders.[br]Although upstream kinase signaling pathways affecting autophagy have been identified, little is known about transcriptional regulation of autophagy. Here we show that in the liver, nutrient status not only controls the initiation of autophagy through the mammalian target of rapamycin (mTOR) complex but also regulates the expression of autophagic genes through nuclear receptors (NRs). Surprisingly, pharmacologic activation of the fasting responsive NR, peroxisome proliferator-activated receptor alpha (PPAR alpha/NR1C1), strongly induces autophagy even in the fed state by enhancing gene expression of autophagic machinery. In contrast, pharmacologic activation of the bile acid NR, farnesoid X receptor (FXR/NR1H4), which reflects the fed state due to enterohepatic bile acid circulation, suppresses autophagy even in the fasting condition by reducing gene expression of autophagic machinery. These specific effects of nuclear receptor agonist ligands are completely lost, as expected, in the relevant PPAR alpha or FXR knockouts. Remarkably, co-treatment with the potent mTORC1 inhibitor Torin1 and PPAR alpha agonist synergistically increases autophagy in murine hepatocytes, suggesting functional coordination of the distinct levels of autophagy regulation. We conclude that nutrient sensing NRs regulate hepatic autophagy at the transcriptional level.[br][br]Nothing to Disclose: JML, MW, RX, DDM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1716 315 2423 MON-545 PO23-02 Monday 2092 2012


2088 ENDO12L_MON-546 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) The Effect of Testosterone Activation of Liver X Receptor on Inflammation and Cholesterol Metabolism and the Implications for Atherosclerosis Treatment Elizabeth Louise Kilby, Thomas Hugh Jones University of Sheffield, Sheffield, UK Low testosterone is associated with an increased prevalence of cardiovascular disease and diabetes and correlates with the degree of atherosclerosis. Testosterone replacement therapy improves several cardiovascular risk factors including the reduction of serum levels of cholesterol and pro-inflammatory cytokines. Liver X Receptor [alpha] (LXR[alpha]), a nuclear receptor is present in various cell types such as macrophages, where it stimulates cholesterol efflux and reduces inflammation. This ability of LXR[alpha] to stimulate the removal of cholesterol from macrophages and suppress their inflammatory response means LXR[alpha] agonists are a potential therapy for atherosclerosis. It was therefore proposed that testosterone acts to reduce the features of atherosclerosis by acting through LXR[alpha]. The human monocyte cell line THP-1, differentiated into macrophages by phorbol 12-myristate 13-acetate was used, as these cells express the androgen receptor and are therefore responsive to testosterone. Differentiated macrophages were treated with 10[sup]-8 [/sup]M testosterone (24, 48 and 72 hr) and gene expression between control and testosterone-treated cells was assessed by qPCR. By 72 hr testosterone significantly increased the expression of [italic]LXR[alpha][/italic] (1.91 with control as 1, p[lt]0.045) and its subtype [italic]LXR[beta][/italic] (1.91, p[lt]0.04) in macrophage cells. In addition, there was a significant increase in the expression of genes downstream of LXR[alpha] which encode proteins involved in cholesterol efflux, including [italic]ABCA1[/italic] ([italic]ATP-binding cassette transporter A1; [/italic]1.92, p[lt]0.03) and [italic]APOE[/italic] ([italic]apolipoprotein E; [/italic]1.98, p[lt]0.05), and [italic]FAS[/italic] ([italic]fatty acid synthase; [/italic]3.26, p[lt]0.001), which produces fatty acids required for the transport and storage of cholesterol. Conversely testosterone treatment was found to downregulate genes encoding the pro-inflammatory cytokines [italic]IL-1[beta] [/italic](0.48, p[lt]0.02),[italic] IL-6 [/italic](0.534, p[lt]0.002) and [italic]TNF[alpha] [/italic](0.285, p[lt]0.0007) by 48 hr. The results suggest testosterone activates LXR[alpha] and acts through this nuclear receptor to control the expression of LXR-target genes such as [italic]ABCA1[/italic], [italic]APOE[/italic], [italic]FAS[/italic], [italic]IL-1B[/italic], [italic]IL-6[/italic] and [italic]TNF[alpha][/italic] to aid cholesterol efflux and suppress inflammation in human macrophages. We therefore hypothesize that testosterone exerts its anti-atherogenic effects in part through the activation of LXR[alpha] and LXR-target genes.[br][br]Nothing to Disclose: ELK, THJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 656 315 2424 MON-546 PO23-02 Monday 2093 2012


2089 ENDO12L_MON-547 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) [beta]-Catenin Negatively Regulates Adipogenesis in 3T3-L1 Cells by Suppression of TR4 Function Hojung Choi, Seung-Jin Kim, Sungsoo Park, Eungseok Kim College of Natural Sciences Chonnam National University, Gwangju, Republic of Korea TR4 is a member of the nuclear receptor superfamily which plays an essential role in energy homeostasis. We recently showed that TR4-deficient mice have reduced white adipose tissues with less lipid accumulation and promoting effect of TR4 on lipid accumulation in adipocytes is in part by increased LCFA uptake. Here we investigated how TR4 facilitates adipogenesis during adipocyte differentiation to explore the role of TR4 in adipocyte biology.[br]We demonstrate here that [beta]-catenin, a negative regulator of adipogenesis, inhibits TR4 activity via upregulation of Slug. Reporter gene assay showed that [beta]-catenin is able to inhibit TR4 transcriptional activity in a dose-dependent manner. Ectopic expression of [beta]-catenin increased Slug mRNA levels and this [beta]-catenin induced expression of Slug resulted in decreased TR4 activity. Furthermore, GST pull-down and mammalian two-hybrid assays showed that Slug physically interacts with TR4. Addition of slug reduced TR4 binding to its cognate response element in the gel shift assay, suggesting that Slug suppresses TR4 activity via inhibition of TR4 binding to TR4 response element located within the promoters of TR4 target genes. Consistent with this data, Slug inhibited expression of TR4 target genes, FATP1 and CD36, in 3T3-L1 preadipocytes during adiocyte differentiation. Suppressive effect of Slug on TR4 activity was further confirmed in 3T3-L1 adipocytes in which Slug repressed TR4 enhancement of lipid accumulation during adipogenesis.[br]These data suggest that [beta]-catenin plays an inhibitory role in adipogenesis through Slug-mediated inhibition of TR4 transcripional activity, leading to reduced lipid accumulation in 3T3-L1 adipocytes.[br][br]Nothing to Disclose: HC, S-JK, SP, EK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 596 315 2425 MON-547 PO23-02 Monday 2094 2012


2090 ENDO12L_MON-548 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) Regulation of TR4 Expression in 3T3-L1 Adipocytes by GSK-3[beta] Sungsoo Park, Seung-Jin Kim, Hojung Choi, Eungseok Kim Chonnam National University, Gwanju, Republic of Korea Adipose tissue stores excess nutrients in the form of triglycerides and mobilizes them in response to nutritional demands. Dysregulation of the metabolic functions of the adipocytes is frequently associated with insulin resistance and type 2 diabetes.[br]Recently, we showed that TR4 promotes lipid accumulation in 3T3-L1 adipocytes via upregulation of fatty acid transporters and TR4-deficient mice show reduced lipid accumulation in white adipose tissues. Therefore, pharmacological control of TR4 expression or its transcriptional activity is of great interest. Here, we show that Glycogen synthase kinase-3[beta] (GSK-3[beta]) induces TR4 expression in 3T3-L1 adipocytes via activation of CREB. Overexpression of GSK-3[beta] in 3T3-L1 cells induced TR4 expression. In contrast, when KCREB, a dominant negative CREB, was overexpressed together with GSK-3[beta] to 3T3-L1 cells, GSK-3[beta] induction of TR4 expression was abolished. Furthermore, inducing effect of low glucose on TR4 expression disappeared when SB415286, a GSK-3[beta] inhibitor, was added to the 3T3-L1 adipocytes. GSK-3[beta] enhanced CREB transactivation of TR4 promoter activity while SB415286, dramatically suppressed CREB-mediated TR4 promoter activity.[br]Together, our results demonstrate that GSK-3[beta] induces TR4 expression in 3T3-L1 adipocytes and this GSK-3[beta] effect at least in part by activation of CREB, suggesting that GSK-3[beta][rarr]CREB[rarr]TR4 establishes the new pathway to modulate metabolic programs in adipocytes.[br][br]Nothing to Disclose: SP, S-JK, HC, EK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 600 315 2426 MON-548 PO23-02 Monday 2095 2012


2091 ENDO12L_MON-549 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) Thyroid Hormone-Mediated Activation of Non-Classical Hepatic Cholesterol Uptake Stephen Ayers, Anusha Angajala, Jan Lammel, Paul Webb Methodist Hospital Research Institute, Houston, TX Thyroid hormone (T3) regulates serum lipid parameters, increasing the hepatic uptake of lipoproteins in various animal models as well as human patients. This effect is mediated by the transcriptional activation of target genes in liver by thyroid hormone receptor beta (THRB), which include various genes involved in the LDL receptor-mediated uptake pathway. Here, we present evidence for the regulation of a new LDL receptor-independent uptake pathway. This uptake is associated with the transcriptional activation of very low density lipoprotein receptor (VLDLR) in liver, an effect observed in hepatocyte cell models and animal models. The regulation of this gene was observed to occur through a set of conserved elements, previously found to mediate constitutive androstane receptor[apos]s (CAR) regulation of this gene in liver, suggesting a functional overlap in the regulatory actions of these receptors. These findings suggest that thyromimetics could provide valuable alternative therapies for the treatment of dyslipidaemias.[br][br]Nothing to Disclose: SA, AA, JL, PW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2322 315 2427 MON-549 PO23-02 Monday 2096 2012


2092 ENDO12L_MON-550 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) Understanding Mechanistic Differences between Thyroid Hormone Receptor [alpha] and [beta] Isoforms: Use of Domain Swap Chymeras Marie Togashi, Aleksandra Cvoro, Rilva Grigorio Pinho Soares, Geilliany Oliveira Campos, Douglas Sieglaff, Paul Webb University of Bras[iacute]lia, Bras[iacute]lia, Brazil; The Methodist Hospital Research Institute, Houston, TX The understanding of structure-function characteristics of different thyroid hormone receptor (TR) isoforms is regarded as an important tool to aid the design of selective thyromimetics. There is a high homology between the two isoforms and the few differences observed in primary sequences account for well-known distinguished responses. Preliminary results using TR alpha and TR beta stable cell lines have shown that TR subtypes activate highly similar gene sets but with wide variations in the magnitude, kinetics/potency of response and response to selective ligands. We performed domain swaps between the two TR isoforms (chymeras) and deletions of the A/B domain in expression vectors to address which regions of the receptor are responsible for differences in TR action on ideal consensus thyroid hormone positively regulated response elements (REs), DR4 and IP6, and a negatively regulated one, TSH alpha. Preliminary results obtained by gene reporter assays in transiently transfected HeLa cells suggest that TR responses on DR4 seem not to be influenced by A/B domain deletion, but that higher levels of TR beta activation at IP6 elements and greater hormone dependency of repression at the TSH alpha element are functions of A/B domains. We are performing a detailed characterization of the TR alpha and TR beta chimeras from cofactor exchange and affinity, transcriptional activation to oligomeric formation on REs to understand the role of each domain in different REs and use them to understand TR action in natural promoters.[br][br]Nothing to Disclose: MT, AC, RGS, GOC, DS, PW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2158 315 2428 MON-550 PO23-02 Monday 2097 2012


2093 ENDO12L_MON-551 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) New Partial Agonists of PPAR[gamma] Potently Suppress IL-1[beta] and Other Pro-Inflammatory Genes in Microphages Joey Z Liu, Paul Webb, Christopher J Lyon, Willa A Hsueh The Methodist Hospital Research Institute, Houston, TX Obesity and type 2 diabetes trigger low-grade systemic inflammation which is now recognized as a key determinant of whole body metabolism and tissue damage. Thiazolidinediones (TZDs), such as rosiglitazone (RSG), are synthetic ligands for peroxisome proliferator activated receptor g (PPARg) and are insulin-sensitizing agents that suppress inflammatory responses. Clinical use of TZDs, however, is severely restricted because of side effects, such as edema, weight gain, bone fractures, and increased risk of heart failure.[br]In this study, we investigated a new class of TZDs (GQ16 and GQ19) for their anti-inflammatory properties in lipopolysaccharide (LPS) treated peritoneal macrophages (PMs) harvested from C57Bl/6 mice. GQ compounds are partial agonists of PPARg and can block Cdk5-mediated phosphorylation of PPARg. Unlike TZDs, GQs only modestly alter H12, the site of co-activator and co-repressor recruitment in the PPAR[gamma] structure. In PMs, GQ16 and GQ19 potently inhibit LPS-induced IL-1b, a major target of approaches currently being tested for treatment of diabetes, demonstrated by real-time PCR (20-fold, p[lt]0.001), ELISA (18-fold, p[lt]0.001), and western blotting (15-fold, p[lt]0.001). In contrast, RSG and a partial agonist MRL24 which inhibits Cdk5 phosporylation, have little effects on LPS-induced expression of IL-1b. None of the drugs inhibited inflammasome proteins. GQs also suppress LPS-induced expression of other pro-inflammatory genes: IL-1a, IL-6, IL-12b, MCP-1, and COX2, with less suppression of TNFa and iNOS, which are classical for RSG, indicating GQs may modulate inflammatory genes through different mechanisms than RSG. MRL24 had little effect on any of the LPS-induced cytokines. In summary, we have identified new partial agonists of PPARg that display more potent anti-inflammatory effects than RSG and MRL24. These alternative TZDs provide a potential for anti-inflammatory applications in obesity and obesity-related complications.[br][br]Nothing to Disclose: JZL, PW, CJL, WAH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2159 315 2429 MON-551 PO23-02 Monday 2098 2012


2094 ENDO12L_MON-552 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) PPAR[gamma] Agonism Promotes the Formation of Nuclear PPAR[gamma]-pERK Complexes That Mediate Cognitive Rescue in TG2576 Alzheimer[apos]s Mice Kelly T Dineley, Jordan B Jahrling, Sigmund J Haidacher, Larry Denner University of Texas Medical Branch, Galveston, TX; University of Texas Medical Branch, Galveston, TX The hippocampus is requisite for the establishment and storage of new memories and is particularly vulnerable to the ravages of Alzheimer[apos]s disease (AD). Clinical trials showed that treatment of early-AD patients with drugs that activate PPAR[gamma] can improve hippocampus-dependent cognitive function. PPAR[gamma] agonism with rosiglitazone (RSG) also rescues cognitive deficits in the Tg2576 AD mouse model. We found novel nuclear PPAR[gamma]-pERK complexes that were elevated in Tg2576 mice with higher cognitive function. Further, these complexes were modulated by RSG feeding and were present in human AD brain. Using qPCR and quantitative mass spectrometry, we found that RSG-mediated cognitive rescue is accompanied by the induction of several PPRE-containing genes and PPAR[gamma]-regulated proteins, many of which are important for mitochondrial and synaptic function including components of the ERK MAPK pathway. ERK MAPK activity (pERK) is required for proper hippocampus-dependent cognitive performance.[br]During hippocampal memory consolidation both untreated and RSG-treated Tg2576 exhibited a significant increase in PPAR[gamma] protein but only RSG-treated Tg2576 exhibited memory consolidation and an increase in nuclear PPAR[gamma]-pERK complexes. Thus, in AD mice, PPAR[gamma] activation by RSG serves a compensatory role by increasing ERK activity in the nucleus during memory consolidation. This compensatory mechanism appears to be specific to AD mice in that PPAR[gamma]-pERK complexes remained unaltered in wildtype animals. RSG promotion of nuclear PPAR[gamma]-pERK complexes in humans is a potential novel therapeutic strategy to control or even prevent AD progression.[br][br]Sources of Research Support: The NIH (R01-AG031859) and the American Health Assistance Foundation to KTD and LD.[br][br]Nothing to Disclose: KTD, JBJ, SJH, LD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1596 315 2430 MON-552 PO23-02 Monday 2099 2012


2095 ENDO12L_MON-553 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) The Roles of PPAR Gamma in Regulation of Adrenal Glomerulosa Cell Functions Ding Xie, Wendy Bollag Georgia Health Sciences University, Augusta, GA Obesity and the related disease of metabolic syndrome are global health epidemics. Aldosterone, the primary mineralocorticoid hormone, has been shown to contribute to the pathogenesis of metabolic syndrome. However, the molecular mechanisms for hyperaldosteronism in obesity and metabolic syndrome remain unknown. Peroxisome proliferator activated receptor-gamma (PPAR[gamma]) is a ligand-activated transcription factor that plays a pivotal role in adipogenesis and other processes as a [ldquo]lipid sensor[rdquo]. In a human adrenocortical carcinoma cell line, H295R, recent data show that a PPAR[gamma] agonist inhibits chronic aldosterone production. In this study, we explored the roles PPAR[gamma] in regulation of aldosterone production in primary bovine adrenal glomerulosa (BAG) cells and in proliferation in HAC15 cells, a clonal derivative of H295R. Pretreatment of BAG cells for 24hours with pioglitazone, a PPAR[gamma] agonist dose-dependently decreased basal aldosterone production. Also, pioglitazone decreased 10nM angiotensin II (AngII)-induced aldosterone production during both the 1[sup]st[/sup] (initial phase) and 2[sup]nd[/sup](sustained phase) hour of AngII stimulation. Even without preincubation, pioglitazone significantly inhibited AngII-induced aldosterone production in the sustained phase. In HAC 15 cells, 10[mu]M pioglitazone significantly inhibited AngII-induced cell proliferation. In addition, microarray analysis performed in HAC15 cells indicated that pioglitazone regulates the expression of several important candidates potentially involved in PPAR[gamma] agonist-mediated suppression of aldosterone production, namely it downregulates angiotensin II receptor, type I and upregulates melanocortin 2 receptor, a receptor for adrenocorticotropic hormone. Taken together, these data indicate that PPAR[gamma] may play a role in regulation of adrenal glomerulosa cell functions and may contribute to obesity-induced hyperaldosteronism to some degree. Furthermore, treatment of the metabolic syndrome and related co-morbidities with PPAR[gamma] agonists may also help to ameliorate hypertension associated with these disorders.[br][br]Nothing to Disclose: DX, WB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2334 315 2431 MON-553 PO23-02 Monday 2100 2012


2096 ENDO12L_MON-554 POSTER SESSION: Orphan [amp] Metabolic Nuclear Receptors (1:30 PM-3:30 PM) [italic]In Vitro[/italic] and [italic]In Vivo [/italic]Screening of Xenobiotics Targeting PPAR[gamma] Using Reporter Cell Lines and GFP-Transgenic Zebrafish ([italic]Danio rerio[/italic]) Larvae Anne Riu, Caroline Pinto, Nicole Ducharme, Abdel Boulahtouf, Catherine McCollum, Patrick Balaguer, Maria Bondesson, Jan-Ake Gustafsson University of Houston, Houston, TX; INSERM-U896, Montpellier, France The peroxysome proliferator-activated receptor [gamma] (PPAR[gamma]) belongs to the nuclear receptor (NR) superfamily composed of 48 members in humans. PPAR[gamma] is known to be a master regulator of adipogenesis, the process of creating adipocytes. Indeed, in 3T3 cell lines, environmental pollutants such as halogenated Bisphenol A, phthalates, tributyltin (TBT) are able to promote adipocyte differentiation through the activation of PPAR[gamma]. The disruption of regulation pathways under the control of this NR may be involved in the onset of metabolic disorders such as obesity leading to insulin resistance, dyslipidemia, type 2 diabetes and cardiovascular disease. Obesity is strongly linked with exposure to risk factors during fetal and infant development. Recently, it has been shown that mice exposed [italic]in utero[/italic] to TBT were predisposed to obesity later in life. Because of the increasing incidence of these metabolic diseases in human and the growing list of chemicals, it is important to develop [italic]in vitro [/italic]and [italic]in vivo[/italic] models in order to screen potential obesogens able to bind and activate PPAR[gamma].[br]In the past decade, zebrafish has been shown to constitute a powerful vertebrate animal model, due to their small size, optical clarity, [italic]ex-utero[/italic] development, fecundity and rapid development. Importantly, embryos and larvae readily absorb/intake compounds from their aqueous environment. With 70% identity, the PPAR[gamma] ligand binding domain (LBD) is evolutionarily conserved between zebrafish and humans. In order to screen for obesogenic compounds, we have generated a stably transfected HGELN-GAL4-zfPPAR[gamma]-LBD reporter cell line and a transgenic GAL4-zfPPAR[gamma]-LBD reporter fish (GAL4-zfPPAR[gamma]-LBD driving the expression of a UAS:GFP, green fluorescent protein, reporter). These two models will allow us to identify and evaluate new obesogens. Moreover, the [italic]in vivo[/italic] zebrafish model will allow us to assess the uptake of compounds, as well as the potency of their biotransformation products to activate PPAR[gamma].[br][br]Nothing to Disclose: AR, CP, ND, AB, CM, PB, MB, J-AG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2129 315 2432 MON-554 PO23-02 Monday 2101 2012


2097 ENDO12L_MON-559 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) A Pitx2 Regulated microRNA Pathway Controls HMGN2 Expression: A Major Regulator of Transcription Factor Activity, Cell Proliferation and Differentiation Jianbo Wang, Xiao Li, Shan Gao, Huojun Cao, Brad A Amendt Texas A[amp]M Health Science Center, Institute of Biosciences and Technology, Houston, TX We have previously shown that HMGN2 forms an inactive transcriptional complex with the Pitx2 homeodomain protein and acetylated chromatin. HMGN2 expression is down-regulated as development proceeds and is not expressed in the neonate mouse. To understand how HMGN2 is developmentally regulated we identified microRNA23a/b (miRNA-23a/b) as targeting HMGN2 expression during development. We use mouse models, cell lines, Pitx2 and HMGN2 knockout and transgenic primary cells, real-time PCR, chromatin immunoprecipitation assay, lentiviral miR expression and inhibitors to analyze the effect of miRNA-23a/b activity on gene expression mechanisms. miRNA-23a/b are highly expressed in oral epithelial and pituitary cells during development and repress endogenous HMGN2 expression. Furthermore, miRNA-23a/b over expression decreases cell proliferation. The HMGN2 null mice are early embryonic lethal and the conditional knockout reveal growth retardation and developmental defects. Pitx2 represses miRNA-23a-27a-24-2 cluster expression in epithelial cells and activates HMGN2 expression. HMGN2 transgenic mice have decreased Pitx2 expression while miRNA-23a/b over expression activates Pitx2 endogenous expression. ChIP assays demonstrate Pitx2 binding to the miRNA-23a/b 5[apos] flanking regions and HMGN2 promoter. HMGN2 regulates the transcriptional activity of multiple transcription factors and appears to be a major regulator of gene expression. HMGN2 is highly expressed in ES cells revealing a role in early developmental processes and inhibition of miRNA-23a/b activity increases cell proliferation in differentiated cells. Therefore, HMGN2 represses the activity of transcription factors required for patterning and differentiation of tissues/cells. These results reveal a role for miRNA-23a/b in regulating HMGN2 and cell proliferation/differentiation. We have identified a novel pathway where Pitx2 activates HMGN2 expression and represses miRNA-23a/b expression. However, HMGN2 feeds back to inhibit Pitx2 transcriptional activity, where it de-represses miRNA-23a/b expression to allow for effective HMGN2 repression during later stages of development. miRNA-23a/b may facilitate cell differentiation by inhibiting HMGN2 expression to allow for transcription factor activation required for early development.[br][br]Sources of Research Support: National Institute of Dental [amp] Craniofacial Research grants DE013941 and DE 018885 awarded to BAA.[br][br]Nothing to Disclose: JW, XL, SG, HC, BAA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2219 316 2433 MON-559 PO16-01 Monday 2102 2012


2098 ENDO12L_MON-560 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Design and Function of a Plasmid-Based microRNA Inhibitor System [italic]In Vitro[/italic] and [italic]In Vivo[/italic]: Knockdown of the miR17-92 Cluster Reveals Multiple Defects Huojun Cao, Xiao Li, Jianbo Wang, Shan Gao, Brad A Amendt Texas A[amp]M Health Science Center, Institute of Biosciences and Technology, Houston, TX microRNAs (miRNAs) are known to control gene expression patterns in cells, tissues and organs and play a role in cancer mechanisms. The recognition of miRNA biological mechanisms has led to development of methodologies to interrupt and inhibit the activities of miRNAs in development and disease. Current knockout and knockdown strategies for the analyses of miRNA function in animal models, cancer and diseases has focused on specific miRNAs using expensive gene targeting mutagenesis or high levels of chemically modified miRNA antagomirs or inhibitors. We have developed a plasmid-based miRNA inhibitor system (PMIS) that effectively knocks down [gt]85% of miRNA activity in cells and mice. The silencing of the miRNA family is dependent on the conserved seed region of the miRNAs and these miRNA inhibitors are highly specific and distinguish between miRNA family members within clusters. Because of the engineered secondary structure and flanking sequences of the miRNA inhibitors they are stable and form a complex with the miRNA, Agonaute and Dicer proteins in a RNA-Induced Silencing Complex (RISC) complex. The miRNA inhibitor is not processed in the RISC complex due to the length of the double stranded regions of the inhibitor required for its interaction with the miRNA. The PMIS was tested in cells and transgenic animals targeting the miRNA 17-92, miRNA 106a-363, miRNA 106b-25 clusters. The mice expressing these inhibitors recapitulated the previously reported miRNA 17-92 knockout mice phenotype, including lung, heart, and craniofacial defects. Furthermore, knockdown of the murine miR-17-92 cluster by our system mimics the skeletal and growth defects observed in human deletions of MIR17HG locus. The miRNA inhibitors are specific for identical miRNA seed sequences in each cluster and plasmids expressing multiple miRNA inhibitors targeting specific seed regions can effectively inhibit the miRNAs of that cluster. These miRNA inhibitors are dependent on their 5[apos] and 3[apos] flanking sequences, double stranded regions, hairpin structures and sequence length that we demonstrate using nucleotide substitutions. These data demonstrate for the first time a new mechanism to inhibit miRNA activity in cells and mice. The PMIS system targets miRNA families to understand their role during development (in vivo) and cell/tissue interactions (in vitro). These new class of microRNA inhibitors can be used as specific miRNA therapeutic reagents in cancer and diseases.[br][br]Sources of Research Support: National Institute of Dental and Craniofacial Research grants DE13941 and DE018885 awarded to BAA.[br][br]Nothing to Disclose: HC, XL, JW, SG, BAA 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2240 316 2434 MON-560 PO16-01 Monday 2103 2012


2099 ENDO12L_MON-561 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Evaluating the Role of Dicer1 in Mouse Thyroid Henriette L Undeutsch, Fuping P Zhang, Pirjo Pakarinen, Matti Poutanen, Jukka Kero University of Turku, Turku, Finland; University of Turku, Turku, Finland microRNAs are small non-coding RNAs that are post-transcriptionally regulating the gene expression. Dicer1 is a key enzyme influencing the microRNA processing. Tissue-specific silencing of Dicer1 provides a tool to study the influence of RNA interference in the tissue development and function. So far, little is known about miRNA in the development and function of the thyroid gland, but several miRNA genes are known to play a role in thyroid cancer.[br]To understand the role of Dicer1 in thyroid in vivo, we have generated a thyroid-specific Dicer1 knockout mouse line by crossing Dicer1 fl/fl mice with Tg-Cre mice expressing the Cre-recombinase under the thyroglobulin promoter.[br]These mice were shown to lack Dicer1 in thyrocytes, and as a consequence, showed an altered histology of the thyroid gland with a decreased formation of thyroid follicles. Furthermore, the mice developed hypothyroidism with a 2-fold increased thyroid stimulating hormone (TSH) bioactivity and 50 % reduced free thyroxine (fT4) serum levels compared to the wild-type mice. The data, thus, indicate that Dicer1 plays a crucial role in thyroid function.[br]In addition, we are in process of generating an inducible thyrocyte-specific Dicer1 KO model by applying a mouse line expressing a tamoxifen-inducible Cre under the thyroglobulin promoter. Characterizing the constitutive and inducible Tg-Cre Dicer1 KO mice will provide us with novel information about the role of Dicer1 in the development and function of the thyroid gland.[br][br]Nothing to Disclose: HLU, FPZ, PP, MP, JK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1517 316 2435 MON-561 PO16-01 Monday 2104 2012


2100 ENDO12L_MON-562 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Germ Cell Nuclear Factor Indirectly Activates Cyclin D1 Expression during Embryonic Stem Cell Differentiation Hongran Wang, Austin J Cooney Baylor College of Medicine, Houston, TX One of the significant markers of embryonic stem cell (ESC) differentiation is increased expression of Cyclin D and the switch from Cyclin D-independent to Cyclin D-dependent regulation of the proliferative cell cycle. However, the molecular mechanism underlying the increased translation of Cyclin D protein remains unknown. Germ Cell Nuclear Factor (GCNF) is a transcriptional repressor that has been shown to regulate early embryonic development and repression of Oct4 expression during ESC differentiation. GCNF expression increases rapidly after embryo implantation. Using GCNF knock out (GCNF KO) embryonic stem cells and embryos, we show that Cyclin D1 is not expressed in the KOs after gastrulation as normal or after differentiation of the GCNF KO ES cells. However, the mRNA is still expressed suggestive of posttranscriptional regulation. Consistently, mir-302, which is predicted to interact with the Cyclin D1 mRNA 3[apos]UTR to repress its translation, was found to be expressed at extremely high levels in GCNF KO ES cells compared wt ES cells. Transfection of mir-302 mimic oligonucleotides inhibits the Cyclin D1 translation that normally occurs in wt ES cell differentiation at day 3 of RA treatment. Inhibiting mir-302 expression in GCNF KO ES cells rescued Cyclin D1 expression. GCNF normally represses mir-302 by directly binding a DR0 response element within the mir-302 promotor. These results demonstrate that GCNF turns on Cyclin D1 expression by inhibiting Mir-302 expression. GCNF is therefore a key regulator for embryo development and for the cell cycle regulation during embryonic development.[br][br]Nothing to Disclose: HW, AJC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2301 316 2436 MON-562 PO16-01 Monday 2105 2012


2101 ENDO12L_MON-563 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Regulatable Expression Systems in Embryonic Stem Cells Qin Wang, Austin J Cooney Baylor College of Medicine, Houston, TX Embryonic stem (ES) cells have the characteristics of pluripotency and self-renewal, that is, they can differentiate into all cell types in the body and can proliferate indefinitely. Study of ES cell mechanisms contributes to potential for regenerative medicine. ES cell pluripotency and self-renewal are controlled by the core pluripotency factors, Oct4, Sox2 and Nanog. The nuclear receptor NR6A1, GCNF, which plays important roles in reproduction and development, binds to the Oct4 proximal promoter and is required for Oct4 repression during retinoic acid (RA) induced differentiation of ES cells. To study ES cell related transcription factor networks, we generated doxycycline regulatable ES cell lines expressing GCNF, the nuclear receptor coregulator SAFB1, the nuclear receptor NR5A2 (LRH1), and the reprogramming factor KLF4. Dox regulated GCNF induction, in undifferentiated LIF treated ES cells did not affect the expression of the pluripotency genes examined. However, induction of GCNF expression in ES cells in the presence of RA led to the repression of many pluripotency genes, such as OCT4, NANOG, SOX2, LRH1, GDF3, NODAL, and REX1. Similarly, Dox regulated SAFB1 induction, in undifferentiated LIF treated ES cells, did not affect the expression of pluripotency genes examined. However induction of SAFB1 expression in the presence of RA led to the repression of OCT4 and NANOG with the similar time course as GCNF, suggesting a possible link. LRH1 induction activated the expression of the pluripotency and endoderm transcription factor TBX3 in all conditions examined. KLF4 induction strongly repressed cell proliferation and viability and enhanced ES cell differentiation in the presence or absence of LIF. In total, these studies contribute to a better understanding of the transcription factor regulatory networks of ES cell pluripotency and self-renewal.[br][br]Nothing to Disclose: QW, AJC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2290 316 2437 MON-563 PO16-01 Monday 2106 2012


2102 ENDO12L_MON-564 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Rapid Change of Nestin-Positive Cells in the Pituitary during the First Postnatal Growth Wave Saishu Yoshida, Hideji Yako, Takako Kato, Yukio Kato Meiji University, Kanagawa, Japan; Meiji University, Kanagawa, Japan; Meiji University, Kanagawa, Japan The anterior pituitary gland consists of five hormone-producing cells and non-hormone producing cells. The latter is composing of folliculo-stellate cells, undifferentiated SOX2-expressing stem/progenitor cells and others. Recently, we reported that a pituitary-specific transcription factor PROP1 is consistently coexists in SOX2-positive undifferentiated cells and disappears at early stage of the transition from progenitor cell to commitment cell during the embryonic development of the rat pituitary (Yoshida[italic] et al.[/italic] 2009). Transition of stem/progenitor cell in the rat postnatal pituitary during the first postnatal growth wave is also reported (Yoshida[italic] et al.[/italic] 2011).[br]To examine further the transition of rat pituitary progenitor cells during the first postnatal growth, we tracked down the cells expressing NESTIN, a marker of multiple stem/progenitor cells. Immunohistochemistry using anti-NESTIN, -PROP1, -PIT1, -SF1, -S100 and -pituitary hormones was performed for rat pituitaries of postnatal day (P) 5, 15 and 30. A ration of NESTIN-positive cells in the cells of anterior lobe was 4, 3 and 3.5 % cells on P5, P15 and P30, respectively. However, it decreased to 0.08% on P70. In the anterior lobe of P5, 90% of the NESTIN-positive cells were PIT1-positive and 13% of those were PROP1-positive. On P15 and P30, PIT1/NESTIN-double-positive cells were around 70% of NESTIN-positive cells, but PROP1/NESTIN-double-positive cell was not detected as well as SF1/NESTIN-double-positive cell. S100/NESTIN-double-positive cells were detected at 1.5 and 4% on P15 and P30, respectively. In the period from P5 to P30, major hormone detected in NESTIN-positive cells was GH (26-33%) and minor one was ACTH (2-9%), LHb (4% on P5 only) and PRL (9% on P30 only).[br]These data suggested that nestin-positive cells outgrow from stem/progenitor cells, transit to commitment cells and develop to terminally differentiate into hormone-producing cells during the first postnatal growth wave in the anterior lobe. Since nestin was rarely observed in anterior lobe of P70, development system might be qualitatively altered after termination of the growth wave.[br][br](1)Yoshida S, Kato T, Susa T, Cai L-Y, Nakayama M and Kato Y: Biochem and Biophys Res Commun, 385: 11-15. 2009. (2)Yoshida S, Kato T, Yako H, Susa T, Cai L-Y, Osuna M, Inoue K and Kato Y: J Neuroendocrinol, 23: 933-943. 2011.[br][br]Nothing to Disclose: SY, HY, TK, YK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1488 316 2438 MON-564 PO16-01 Monday 2107 2012


2103 ENDO12L_MON-565 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Expression of Stem Cell Pluripotency Factors in Intestine Remodeling during Metamorphosis in [italic]Xenopus laevis[/italic] Yu Zhang, Yunbo Shi National Institutes of Health, Bethesda, MD [italic]Xenopus laevis[/italic] intestine is extensively remodeled during the metamorphosis. There is a similar dedifferentiation process between dedifferentiation from somatic cells to induced pluripotent stem cells in mammals and that in the natural metamorphosis in [italic]Xenopus laevis[/italic]. To investigate if mammalian stem cell pluripotency factors were involved in the intestine remodeling in [italic]Xenopus laevis[/italic] metamorphosis, the expression of klf4, sox2, c-myc, oct25, oct60, oct79 (homologues of oct4) were tested during the natural metamorphosis, thyroid hormone-induced metamorphosis and tissue-specific metamorphosis. We found that although all these pluripotency factors, klf4, sox2, c-myc, oct25, oct60, oct79 (homologues of oct4), were all expressed during intestine remodeling in metamorphosis, they might not regulate intestine remodeling in [italic]Xenopus laevis[/italic] metamorphosis.[br][br]Nothing to Disclose: YZ, YS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2188 316 2439 MON-565 PO16-01 Monday 2108 2012


2104 ENDO12L_MON-566 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) [italic]O[/italic]-Glucose Regulation of Notch Signaling in Mammary Gland Morphogenesis and Tumorigenesis Rodrigo Fernandez-Valdivia, John P Lydon, Hamed Jafar-Nejad Institute of Molecular Medicine, The University of Texas Health Science Center, Houston, TX; Baylor College of Medicine, Houston, TX; The University of Texas Health Science Center, Houston, TX; The University of Texas Health Science Center, Houston, TX; Baylor College of Medicine, Houston, TX Notch signaling in mammary morphogenesis and tumorigenesis was first recognized when a frequent common insertion site (INT3) for the mouse mammary tumor virus (MMTV) was mapped to the [italic]Notch4[/italic] gene. This insertional event triggers expression of a constitutively active form of Notch4, causing mammary tumor formation. Moreover, transgenic overexpression of activated forms of [italic]Notch1[/italic], [italic]3[/italic], and [italic]4[/italic] individually is sufficient to cause mammary tumorigenesis. Elevated expression of Notch1 and Jagged1 in mammary tumors correlates with poor prognosis, whereas downregulation of negative regulators of Notch signaling is consistently observed in these tumors. Both Notch and Wnt pathways regulate mammary gland stem cell (MaSC) homeostasis. Indeed, Notch signaling is required for Wnt1-induced transformation of primary human mammary epithelial cells both in vitro and in vivo, indicating that a cross-talk between these pathways is required for breast cancer progression. In addition, the Notch pathway is upregulated in both human and [italic]MMTV-Wnt1[/italic] murine breast cancer stem cells. We have recently discovered Rumi, the sole [ldquo]protein [italic]O[/italic]-glucosyltransferase[rdquo] (Poglut1) present in mammals. Rumi regulates Notch signaling through [italic]O[/italic]-glucosylation of specific serine residues in some EGF repeats of Notch receptors, and acts as a general regulator of the pathway. Interestingly, even removing one copy of [italic]Rumi[/italic] significantly affects Jagged1-induced signaling in mice. Our data indicate that Rumi is highly expressed in the nulliparous mammary epithelium, a cellular compartment that harbors the MaSC population, and that Rumi regulates the Notch pathway in breast cancer cells in a protein level-dependent manner. We will also show our results on the regulation of Rumi expression at different stages of pregnancy, and Rumi expression analysis in progesterone-dependent and Wnt pathway activation-dependent mammary gland tumors. Finally, we will present our strategy to ablate Rumi in the distinct cellular compartments of the mammary gland, including the MaSC population. If a genetic decrease in Rumi can halt the progression of breast cancer in mice, pharmacological targeting of this new enzyme could serve as a novel therapeutic approach to prevent breast cancer relapse by inhibiting Notch signaling in breast cancer stem cells.[br][br]Nothing to Disclose: RF-V, JPL, HJ-N 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2238 316 2440 MON-566 PO16-01 Monday 2109 2012


2105 ENDO12L_MON-567 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Uterine DNA Methylation Regulates Stromal Cell Decidualization Fei Gao, Xinghong Ma, Allison Rusie, Jennifer Hemingway, Alicia B Ostmann, Daesuk Chung, Sanjoy K Das Cincinnati Children[apos]s Hospital Medical Center, Cincinnati, OH; Cincinnati Children[apos]s Hospital Medical Center, Cincinnati, OH; Northeast Agricultural University, Harbin, China Embryo-uterine interaction during early pregnancy critically depends on the expression of numerous genes at the site of implantation. The epigenetic mechanism that occurs through DNA methylation (DNM) plays a major role in the control of gene expression, although this regulatory event remains unknown in the uterus during early pregnancy. To elucidate this notion, we first examined the expression of DNA methyltransferases (Dnmts) and 5-methycytosine (5-mC) in the periimplantation uteri of mice. Dnmt1 and Dnmt3a were absent on day 1 (D1) of pregnancy. In contrast, Dnmt1 was predominantly detected in endometrial cells on the receptive (D4) and early post-attachment (D5) phases, while Dnmt3a had poor localization with somewhat patchy distribution. Interestingly, both Dnmt1 and Dnmt3a were coordinately expressed in decidualizing stromal cells on D6-8. Analysis of 5-mC revealed an expression that was similar to that of Dnmt1 on these days. The preimplantation inhibition of DNM by 5-aza-2[apos]-deoxycytodine (5-aza) was not antagonistic for the embryonic attachment by blue reaction and marker expression, although uterine endometrial stromal cell proliferation at the sites of implantation was significantly downregulated, indicating a disturbance with the post-attachment decidualization event. Indeed, the peri- or post-implantation inhibition of DNM caused significant abrogation of decidualization. This was further consistent with the 5-aza injection studies in experimentally induced decidualization. We next identified decidual genes that undergo alteration of DNM using the methylation-sensitive restriction fingerprinting (MSRF) technique. One such gene, Cbx4, an epigenetic regulator in the polycomb group (PcG) protein family, exhibited hypomethylation in the promoter and increased expression with the onset of decidualization. Furthermore, because uterine Hoxa-10 undergoes alteration of DNM in different biological contexts, and is involved in decidualization, we further identified decidual genes that showed alteration of DNM downstream of Hoxa-10 by utilizing the Hoxa-10 null mice. One such gene, B4galt1, a biosynthesis regulator of different glycoconjugates, exhibited promoter-specific hypermethylation with down-regulated expression in Hoxa-10 null mice, as compared to that of wild-type. Overall, these results suggest that uterine DNM plays a major role for successful decidualization at the site of implantation.[br][br]Sources of Research Support: NIH grants HD056044 and ES007814.[br][br]Nothing to Disclose: FG, XM, AR, JH, ABO, DC, SKD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1632 316 2441 MON-567 PO16-01 Monday 2110 2012


2106 ENDO12L_MON-568 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Epigenetic Control of [italic]FOXE1[/italic] Expression in Humans Rasha Abu-Khudir, Fabien Magne, Johnny Deladoey Endocrinology Service and Research Center, Sainte-Justine University Hospital Center, University of Montr[eacute]al, Montr[eacute]al, Canada; University of Montr[eacute]al, Montr[eacute]al, Canada [bold]Background:[/bold] The forkhead box E1 [italic](FOXE1)[/italic], formerly called thyroid transcription factor 2 [italic](TITF2)[/italic], is a member of the forkhead/winged-helix family of transcription factors. [italic]FOXE1[/italic] is part of an integrated regulatory network of transcription factors that initiate differentiation of the thyroid gland and is essential for its migration. Aberrant methylation of [italic]FOXE1[/italic] has been reported in cancers of the skin, pancreas and breast. In this study, we aimed to determine the role of DNA methylation in regulating the differential expression of [italic]FOXE1[/italic] in non-tumoral tissues, specifically thyroid and leucocytes.[br][bold]Methods:[/bold] Bisulfite sequencing, subsequently validated by bisulfite pyrosequencing, and semi-quantitative RT-PCR were employed to analyze the methylation status of the predicted CpG islands and to compare [italic]FOXE1[/italic] gene expression between human thyroids and matched leucocytes. In addition, [italic]in vitro[/italic] methylation and luciferase reporter assays were used to detect the impact of DNA methylation on [italic]FOXE1[/italic] gene expression in the rat thyroid cell line PC Cl3.[br][bold]Results:[/bold] Bisulfite sequencing revealed that the methylation state of two consecutive CpG dinucleotides (CpG[sub]14[/sub] and CpG[sub]15[/sub]) in CpG island 1 of the 5[apos]-upstream regulatory region of [italic]FOXE1[/italic] (-1600 to -1140 from the transcription start site) is significantly higher in leucocytes than in thyroid tissues (51% for CpG[sub]14[/sub] and 43% for CpG[sub]15[/sub] in leucocytes vs 5% for both CpGs in thyroids; P[lt]0.001). [italic]FOXE1[/italic] expression was found to be inversely associated with the methylation status of this differentially methylated region. Indeed, [italic]in vitro[/italic] methylation of constructs containing different lengths of the 5[apos]-upstream regulatory region of [italic]FOXE1[/italic] showed a strong transcriptional repression of the luciferase reporter in PC Cl3 cells (luciferase activities were 1.5-6.9% of the mock-methylated constructs; P[lt]0.001). [bold]Conclusion:[/bold] Our data indicate that DNA methylation at only two CpG dinucleotides in the 5[apos]-upstream regulatory region of [italic]FOXE1[/italic] may account for its differential expression in normal thyroid and leucocytes. The role of this differentially methylated region in thyroid development and oncogenesis deserves further study.[br][br]Nothing to Disclose: RA-K, FM, JD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 403 316 2442 MON-568 PO16-01 Monday 2111 2012


2107 ENDO12L_MON-569 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Chromatin Activation of Regulatory Elements of the [italic]Igf1[/italic] Gene Is Tissue-Specific Dennis J Chia, Mahalakshmi Santhanam Mount Sinai School of Medicine, New York, NY The diverse roles of IGF -1 in physiology include acting as the endocrine intermediate to elicit the anabolic actions of growth hormone (GH). In previous work, we have begun to characterize the chromatin landscape of the hepatic [italic]Igf1[/italic] gene using a hypophysectomized rat model, and demonstrated that both promoters are enriched for acetylation of histones H3 and H4 and H3K4me3. Here, we assess whether the chromatin landscape of regulatory elements of the [italic]Igf1[/italic] gene characterized in the liver is tissue-specific. We prepared chromatin from liver ([italic]L[/italic]) and spleen ([italic]S[/italic]) of pituitary-intact young adult male C57BL/6J mice for use in FAIRE and ChIP assays. In both liver and spleen, we observed FAIRE enrichment at the promoter of the ubiquitously expressed [italic]Polr2a[/italic] gene that encodes a subunit of RNA Polymerase II ([italic]L[/italic] 4.7X- vs. [italic]S[/italic] 3.9X enrichment relative to a set of 4 negative control segments). We next demonstrated tissue-specificity of FAIRE enrichment at the promoters for [italic]Alb[/italic] encoding albumin ([italic]L[/italic] 12.5X- vs. [italic]S[/italic] 1.3X) and [italic]Ptprc[/italic] encoding CD45 which is specifically expressed in hematopoietic cells ([italic]L[/italic] 1.4X vs. [italic]S[/italic] 10.3X). The [italic]Igf1[/italic] gene is known to be expressed in many tissues, however it is most abundantly expressed in the liver which contributes the vast majority of serum IGF-1. We find FAIRE enrichment at [italic]Igf1[/italic] Promoter 1 is significantly more robust in the liver than the spleen ([italic]L[/italic] 11.9X vs. [italic]S[/italic] 2.3X). Results with both H3K4me3 and H3K9ac-ChIP mirror the FAIRE experiments with liver-specific enrichments at the [italic]Igf1[/italic] and [italic]Alb[/italic] promoters and spleen-specific enrichments at the [italic]Ptprc[/italic] promoter. In additional studies, we evaluated the set of 7 described Stat5-binding domains of the [italic]Igf1[/italic] gene and determined that all lie within regions of open chromatin in liver (range 3.1-23.8X, average 14.0) as assessed by FAIRE, whereas they are in an inaccessible state in spleen (range 0.6-1.4X, average 0.9). Pilot experiments following acute systemic GH did not alter the levels of FAIRE enrichment at the [italic]Igf1[/italic] gene regulatory elements, suggesting that open chromatin at the locus is independent of GH and transcriptional activation, analogous to descriptions of chromatin accessibility pre-determining binding of other transcription factors. Our data support the hypothesis that the [italic]Igf1[/italic] gene specifically acquires marks of chromatin activation during the normal development and differentiation of the liver, and provide a mechanism whereby epigenetic alterations at the [italic]Igf1[/italic] locus can manifest as GH resistance.[br][br]Nothing to Disclose: DJC, MS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1725 316 2443 MON-569 PO16-01 Monday 2112 2012


2108 ENDO12L_MON-570 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Evidence of Germline Epimutations Associated with Diminished MLH1 and MSH2 Expression in Non-Obstructed Azoospermic Men Alex David Ridgeway, Dolores Lamb Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX The ability to produce progeny is a trait that is required for the survival of all mammalian species. As such, infertility is considered by [quot]nature[quot] to be a genetically lethal condition. The advent of assisted reproductive techniques, specifically intracytoplasmic sperm injection (ICSI), has aided many couples with severe male factor infertility to achieve a pregnancy, essentially overcoming nature[apos]s block to fertilization by defective gametes. Currently, ICSI births account for a significant number of children worldwide. There are concerns, however, that the use of ICSI for men with non-obstructed azoospermia (NOA) may have implications for the offspring ranging from birth defects to inherited systemic defects. Studies within our lab suggest that an intrinsic defect in DNA replication and repair via abnormal or absent expression of hMLH1 and hMSH2 may be an underlying cause of NOA in some men (1, 2). Indeed, some of these mutations are those found in Hereditary Non-Polyposis Colorectal Cancer (HNPCC). Therefore, elucidation of the molecular basis of mismatch repair deficiencies in NOA men may better help assess the risks involved in ICSI and its consequences for their offspring. Hypermethylation of the MLH1 promoter downregulates MLH1[apos]s expression in HNPCC (3). We hypothesize that similar germline epigenetic alterations may be associated with MLH1 and MSH2 deficiencies observed in NOA men. The DNA methylation signature of MLH1 and MSH2 from testicular biopsies of NOA patients was analyzed using bisulfite conversion and Methylation-Specific PCR (MSP). Results indicate the presence of methylated CpG sites within the promoter region of MLH1 of some NOA patients (5/23). Of these patients, 3 exhibited methylated CpG sites in the most proximal promoter region that correlates with the absence of MLH1 expression in HNPCC (4). Furthermore, none of the fertile control men examined (0/14) exhibited methylated CpG sites in this proximal region. Epigenetic alterations in mismatch repair gene expression, independent of gene mutations, may provide a novel mechanism in copying a disease phenotype when coupled with ICSI. If methylation of the MLH1 promoter is present in both the somatic and testis tissue of NOA men with absent MLH1 expression, this germline event potentially can be transmitted to the offspring conceived by ICSI.[br][br](1) Mukherjee, S., et al., Curr Opin Urol. 20(6): p. 525-32. (2) Maduro, M.R., et al., Mol Hum Reprod, 2003. 9(2): p. 61-8. (3) Gazzoli, I., et al., Cancer Res, 2002. 62(14): p. 3925-8. (4) Deng, G., et al., Cancer Res, 1999. 59(9): p. 2029-33.[br][br]Sources of Research Support: NIH grant PO1HD36289 to DJL.[br][br]Nothing to Disclose: ADR, DL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 832 316 2444 MON-570 PO16-01 Monday 2113 2012


2109 ENDO12L_MON-571 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Differential DNA Methylation of Human CYP11B1 and CYP11B2 Promoters in Association with Adrenal Zonation Masashi Demura, Fen Wang, Takashi Yoneda, Mitsuhiro Kometani, Toshitaka Sawamura, Yuan Cheng, Katsuhiko Ono, Yasuhiro Nakamura, Hironobu Sasano, Kiyofumi Saijoh, Yoshiyu Takeda Kanazawa University, Kanazawa, Japan; Dept of Pathology, Sendai, Japan; Kanazawa University, Kanazawa, Japan [bold]BACKGROUND:[/bold] Aldosterone and cortisol synthesis are catalyzed by two similar but different CYP11B enzymes, CYP11B2 and CYP11B1, respectively. The CYP11B2 and CYP11B1 genes are located in tandem on chromosome 8, sharing 95% identity in the coding regions and 90% identity in introns. These two genes, however, have little similar upstream sequences and display different zonal expression patterns. In contemporary humans, CYP11B2 was sporadically expressed in the zona glomerulosa (ZG), whereas CYP11B1 was entirely detected in the zona fasciculata (ZF)/reticularis (ZR) and dominantly expressed in ZF. CYP11B1 expression is relatively high compared with CYP11B2. The CYP11B2 and CYP11B1 gene promoters possess a number of target sites for DNA methylation that is closely associated with gene silencing.[br][bold]METHODS and MATERIALS:[/bold] Adrenals from 5 autopsy subjects were assessed for immunohistrochemically defined adrenal zonation, using antibodies for 17alpha-hydroxylase/17,20-lyase (CYP17), 3beta-hydroxysteroid dehydrogenase (HSD3B), and dehydroepiandrosterone-sulfotransferase (SULT2A1). CYP17, HSD3B, and SULT2A1 are entirely expressed in ZF/ZR, ZG/ZF, and ZR, respectively. We used laser capture microscopy to isolate DNA from each zone in adrenal tissues and performed an analysis of DNA methylation. Promoter methylation patterns of the CYP11B2 and CYP11B1 genes in adrenal zones were examined by means of bisulfite sequencing.[br][bold]RESULTS:[/bold] The CYP11B1 gene was predominantly unmethylated in ZF while completely methylated in both ZG and ZR. In contrast, the CYP11B2 gene was hypomethylated in ZG compared with those in ZF and ZR. Inter-individual diversity was observed in DNA methylation of the CYP11B2 gene in ZR. The DNA methylation patterns appeared to exert zone-specific expression patterns of CYP11B2 and CYP11B1 enzymes in conjunction with transcription factors.[br][bold]CONCLUSIONS:[/bold] In terms of expression site and strength, promoter methylation patterns of the CYP11B2 and CYP11B1 genes appeared to reflect a capacity to express CYP11B enzymes. DNA methylation patterns of the CYP11B1 and CYP11B2 promoters were closely associated with the adrenal zonation. Since the expression of aldosterone synthase depends on salt intake, incomplete DNA hypomethylation of the CYP11B2 gene in ZG is ascribed to high-sodium diets in contemporary society.[br][br]Nothing to Disclose: MD, FW, TY, MK, TS, YC, KO, YN, HS, KS, YT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1443 316 2445 MON-571 PO16-01 Monday 2114 2012


2110 ENDO12L_MON-572 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Transcriptional Regulation of Human FDXR Gene by the Transcription Factor SF-1 Yoshitaka Imamichi, Tetsuya Mizutani, Yunfeng Ju, Takehiro Matsumura, Takashi Yazawa, Shinya Kawabe, Masafumi Kanno, Akihiro Umezawa, Kaoru Miyamoto University of Fukui, Fukui, Japan; University of Fukui, Fukui, Japan; University of Fukui, Fukui, Japan; National Institute for Child Health and Development, Tokyo, Japan Steroidogenic factor-1(SF-1, also called Ad4BP) is a master regulator of steroidogenesis which is essentail for development of adrenal gland and gonads. We have shown that steroidogenic factor (SF-1) can induce the differentiation of bone marrow-derived mesenchymal stem cells (MSCs) into steroidogenic cells with the aid of cAMP stimulation. To elucidate the molecular mechanisms that control the differentiation from hMSC into steroidogenic lineage, we analyzed gene expression pattern induced by SF-1 with expression microarrays, and estimated genome-wide binding regions for SF-1 with ChIP-on-Chip. We identified ferredoxin reductase (FDXR) gene as a possible novel target of SF-1, which is essential for the enzyme activity of mitochondrial cytochrome p450 emzymes in steroidogenic cells. Conventional ChIP and EMSA analysis revealed that a novel SF-1 binding region was found near the transcription start site of FDXR gene. Luciferase reporter assay using the reporter constructs containning the newly identified SF-1 binding region revealed that this region was critical for the transription of FDXR gene. Moreover, Overexpression of SF-1 promotes the mRNA expression of FDXR in Hela and a human ovarian granulosa-like tumor cell line, KGN. SF-1 knockdown consistently reduced FDXR mRNA and protein levels in a human adrenal carcinoma cell line, H295R. These results indicate that FDXR participates as a target of SF-1 in metabolic events of steroidogenic tissues.[br][br]Nothing to Disclose: YI, TM, YJ, TM, TY, SK, MK, AU, KM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1171 316 2446 MON-572 PO16-01 Monday 2115 2012


2111 ENDO12L_MON-573 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Mitochondria-to-Nucleus Transcriptional Cross-Talk: Star Protein Induces Upregulation of Mitochondrial Protease Expression Assaf Bahat, Naomi Melamed-Book, Eli Anuka, Joseph Orly The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel StAR activity is instrumental for efficient steroidogenesis in cells of the adrenal cortex and the gonads due to its role in providing a flux of cholesterol substrate to the first and key steroidogenic enzyme CYP11A1/P450scc residing in the inner mitochondrial membranes. Recent studies have shown that StAR is active while interacting with proteins of the outer mitochondrial membrane, hence its import into the matrix serves to terminate this activity. We have shown that a rapid accumulation of StAR in the mitochondrial matrix can generate a cytotoxic stress we named as [apos]protein-overload stress[apos] (POS); our past and present studies show that to seemingly avoid detrimental consequences of POS, the mitochondria rapidly degrade StAR by their autonomous quality control machinery including a sequential action of the the matrix protease Lon, followed by proteolysis at the matrix face of the inner-membrane executed by the mAAA ATP-dependent metalloproteinase AFG3L2 (L2).[br]We further questioned whether StAR accumulation in the mitochondria also signals for enrichment of the mitochondrial protease content aiming to keep POS under control. First, in vivo data support this notion by showing that during ovarian follicular development, L2, LON and HSP60 are selectively augmented in steroidogenic StAR expressing cells. More direct is the finding showing that expression of recombinant wild-type StAR in HEK293 cells led to up to 5-fold increase of L2 over control levels of the protease in cells expressing a non-importable mutant of StAR, N47-StAR. Accumulation of wild-type StAR also signaled a marked elevation of transcription noted by up to 3-fold more CAT reporter activity directed by the promoters of the major mitochondrial proteases involved in StAR degradation, i.e., AFG3L2, SPG7 (L2 chaperon) and LON.[br]Finally, we screened for possible mitochondrial events downstream of StAR accumulation that could mediate the mitochondria-to-nucleus effect on gene-specific transcription. Interestingly, siRNA madeiated knock-down of CLPP protease suppressed 75% of the StAR induced effect on LON transcription; noteworthy is the fact that CLPP, which we previously found unable to degrade StAR, was recently found essential for mediating a mitochondrial unfolded response in c. elegans model experiments. Collectively, these findings suggest a novel mechanism controlling house-keeping genes encoding mitochondrial proteases in steroidogenic cells.[br][br]Nothing to Disclose: AB, NM-B, EA, JO 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 678 316 2447 MON-573 PO16-01 Monday 2116 2012


2112 ENDO12L_MON-574 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Potent Anti-Apoptotic Role for Extensive StAR Induction Revealed in Cardiac Myofibroblasts Following Experimental Infarction in Murine Heart Eli Anuka, Natalie Yivgi-Ohana, Sarah Eimerl, Naomi Melamed-Book, Edith Hochhauser, Joseph Orly The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel; Felsenstein Medical Research Center, Tel Aviv University, Rabin Medical Center, Petah Tikva, Israel Steroidogenic acute regulatory protein, StAR, is indispensable for high output steroid hormone synthesis in cells of the classical steroidogenic tissues. We discovered that high expression of the Star gene is also activated in non-steroidogenic fibroblasts of mouse heart following experimental myocardial infarction (MI). Western analyses, RT-PCR and immunohistochemical evidence show that 3 days after MI, StAR expression rises in fibroblasts (vimentin positive) congregating in the damaged myocardium of the left ventricle outer wall, where no expression of CYP11A1 or 3[beta]HSD was detected. These observations suggest that StAR expression is not coupled to local de novo steroidogenesis. Neonate or adult rat heart fibroblasts put to culture (but not 3T3 cells or MEFs), express high levels of StAR in response to a brief exposure to pro-apoptotic stimuli, such as H2O2 or staurosporine. Such StAR positive fibroblasts do not succumb to apoptosis readily noted in StAR negative cells undergoing nuclear fragmentation. A strong anti-apoptotic impact conferred by StAR was also typical of cardiac fibroblasts terminally differentiating to become [alpha]-smooth muscle actin (SMA) positive cells, i.e. myofibroblasts.[br]Quantitative confocal evidence showed that StAR expression in heart fibroblasts reduces the incidence of induced apoptosis (fragmented nuclei) by nearly 75%. Conversely, siRNA mediated knockdown of StAR to 40% doubled the number of Annexin V positive cells assessed by FACS, reassuring the protective role of StAR against apoptosis. Similar results were obtained with ovarian granulosa cultures treated with staurosporine, or in HeLa cells where recombinant StAR prevented mitochondrial cytochrome c release and nuclear fragmentation.[br]Collectively, these findings revise our understanding of StAR biology suggesting that: (a) high level StAR expression is not necessarily restricted to steroidogenic cells; (b) in all cells, steroidogenic or not, StAR can play the role of an anti-apoptotic survival factor; (c) one example of such are heart myofibroblasts that express StAR in response to stress insults applied prior or following terminal differentiation; (d) finally, cardiac StAR expression is probably of a primary physiological importance since it seems to spare the myofibroblast precursor cells during infarct and paves the way for initiation of a post-MI myocardium tissue repair process, typically governed by myofibroblasts.[br][br]Nothing to Disclose: EA, NY-O, SE, NM-B, EH, JO 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 180 316 2448 MON-574 PO16-01 Monday 2117 2012


2113 ENDO12L_MON-575 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Oct1 Trans-Activates [beta]-Casein Gene Expression through Physical Interactions with STAT5 and GR in Mammary Epithelial Cell Xi Qian, Feng-Qi Zhao The University of Vermont, Burlington, VT [beta]-Casein is a major milk protein synthesized in the mammary gland during lactation. STAT5 (Signal Transducers and Activators of Transcription 5) and GR (glucocorticoid receptor) play essential roles in mediating the induction of [beta]-casein gene expression by prolactin and glucocorticoid in mammary epithelial cell. In addition, we have identified an octamer binding site in the proximal promoter region of the mouse [beta]-casein gene and found that mutations at this site significantly diminish the hormonal induction of the promoter activity. However, little is known about the underlying mechanisms. In this study, chromatin immunoprecipitation showed that Oct1, an ubiquitously expressed octamer-binding transcription factor, binds to [beta]-casein gene promoter [italic]in vivo[/italic] and this binding activity could be induced by lactogenic hormones within 30 min. Oct1 over-expression in mammary epithelial cells resulted in a [sim]50% stimulation of endogenous [beta]-casein protein expression in response to lactogenic hormones while Oct1 knockdown resulted in a [sim]65% inhibition. Lactogenic hormones did not affect Oct1 expression in both mRNA and protein levels. Oct1 was mainly localized in the nuclei of the cells and its subcellular localization was not influenced by lactogenic hormones. Immunoprecipitation experiments demonstrated that lactogenic hormones rapidly induce the physical interactions of Oct1 with both STAT5 and GR on [beta]-casein gene promoter. Finally, in transient transfection experiment, Oct1 synergistically transactivated the [beta]-casein gene promoter activity with STAT5 and GR in the presence of lactogenic hormones. In summary, these findings suggest that Oct1 functions as a transactivator in prolactin and glucocorticoid-induced [beta]-casein gene expression by direct interactions with STAT5 and GR.[br][br]Nothing to Disclose: XQ, F-QZ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 982 316 2449 MON-575 PO16-01 Monday 2118 2012


2114 ENDO12L_MON-576 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Efficient, Specific, Developmentally Appropriate Cre-Mediated Recombination in Anterior Pituitary Gonadotropes and Thyrotropes Maria Ines Perez Millan, Sally A Camper, Shannon W Davis University of Michigan, Ann Arbor, MI Tissue-specific expression of [italic]cre[/italic] recombinase is a well-established genetic tool to analyze gene function, and it is limited only by the efficiency and specificity of available [italic]cre[/italic] mouse strains. Here we report the generation of a transgenic line containing a [italic]cre[/italic] cassette with codon usage optimized for mammalian cells (1) under the control of a mouse glycoprotein hormone [alpha]-subunit ([alpha]GSU or [italic]Cga[/italic], chorionic gonadotropin alpha) regulatory sequences in a bacterial artificial chromosome (BAC) genomic clone. These transgenic mice produce efficient and specific [italic]cre[/italic]-mediated recombination in gonadotrope and thyrotrope cells of the anterior pituitary gland with appropriate developmental onset.[br]Pituitary deletion of transcription factors and signaling pathways was achieved with a smaller [italic]Cga[/italic] transgene (2, 3, 4, 5). This transgenic line, [italic]Tg(Cga-cre)[/italic][sup]3Sac[/sup], has only 4.6 kb of mouse [italic]Cga[/italic] promoter sequences, and its applicability was limited by ectopic activity in skeletal and cardiac muscle. To generate transgenic mice that more accurately recapitulate endogenous [italic]Cga[/italic] gene expression we used homologous recombination in [italic]E. coli[/italic] to introduce the improved [italic]cre[/italic] cassette ([italic]icre[/italic]) into a 228 kb mouse [italic]Cga[/italic] genomic BAC clone in exon 1 at the translation start site. The engineered BAC was used to generate transgenic mice: [italic]Tg(Cga-icre)[/italic].[br]We used several different [italic]cre[/italic]-reporter transgenic mice to assess the efficiency and specificity of the [italic]Tg(Cga-icre)[/italic] BAC transgenic lines. A [italic]cre[/italic]-activated lacZ reporter (R26R) revealed expression in the developing pituitary gland at embryonic day 12.5 in the appropriate pattern. Using the same reporter [italic]Tg(Cga-icre)[/italic] activity was detected in the pituitary of 2 mo old adult mice, with little or no activity in inappropriate tissues such as muscle, brain, kidney, lungs, testis, ovaries, tail and liver. A [italic]cre[/italic]-activated yellow fluorescent protein (YFP) reporter line revealed YFP co-localization with LH[beta], FSH[beta] and TSH[beta] immunoreactive cells, but not with ACTH, PRL, and GH, indicating appropriate pituitary-cell specific restriction.[br]In conclusion, we generated an improved [italic]Tg(Cga-icre)[/italic] BAC transgenic line that targets pituitary thryrotropes and gonadotropes efficiently and specifically. This tool is an important addition to the armament of [italic]cre[/italic] strains for targeting other pituitary cell types (6, 7, 8). BAC recombineered transgenes are an effective alternative to knock-in alleles that can create haploinsufficiency for the endogenous gene.[br][br](1) Shimshek et al. Genesis 32: 19, 2002. (2) Cushman et al. Genesis 28: 164, 2000. (3) Charles et al., Mol Endo 20:1366, 2006. (4) Zhao et al., Development 128:147, 2001. (5) Bliss et al., Mol Endocrinol. 23:1092-101, 2009. (6) Charles et al., Genesis. 46:507-14, 2008. (7) Nasonkin et al., Genesis. 47:55-60, 2009. (8) Castinetti et al., Mol Endocrinol. 25:1950-60, 2011.[br][br]Sources of Research Support: NIH R01HD34283.[br][br]Nothing to Disclose: MIPM, SAC, SWD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1308 316 2450 MON-576 PO16-01 Monday 2119 2012


2115 ENDO12L_MON-577 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Comparison of Gene Expression Profiles of the Human Adrenal Zona Fasciculata and Zona Reticularis Juilee Rege, Tao Wang, Todd D Merchen, William E Rainey Georgia Health Sciences University, Augusta, GA; Georgia Health Sciences University, Augusta, GA [bold]Introduction:[/bold] The human adrenal cortex is anatomically and functionally divided into three concentric zones; the zona glomerulosa (ZG), zona fasciculata (ZF) and zona reticularis (ZR). Mineralocorticoids and glucocorticoids are secreted by ZG and ZF respectively, whereas the human ZR is considered the primary site of adrenal androgen production. However, the exact molecular mechanisms leading to the functional phenotype of the ZF and ZR are still not clearly defined. In the present study, we have identified the differentially expressed genes in the ZF and ZR. [bold]Objective:[/bold] To compare the mRNA transcript profiles between the ZF and ZR. [bold]Materials and Methods:[/bold] ZF and ZR were microdissected from ten human adrenals. Total RNA was extracted from six ZF/ZR pairs and hybridized to an Illumina microarray chip to identify the transcript differences. The most highly expressed transcripts (six ZF-dominant and five ZR-dominant) with biological potential were selected for confirmation with real time quantitative RT-PCR (qPCR). [bold]Results:[/bold] Microarray results demonstrated that the ZF had 139 transcripts with [ge] 2-fold increase compared to its paired ZR. ZR was found to have 65 transcripts with [ge] 2-fold higher expression than in ZF. Microarray and qPCR analysis of transcripts encoding steroidogenic enzymes (N=13) demonstrated that only HSD3B2 (type 2 3beta-hydroxysteroid dehydrogenase) showed higher expression in ZF compared to ZR (P[lt]0.01). In contrast, AKR1C3 (type 5 17beta-hydroxysteroid dehydrogenase) and CYB5 (cytochrome b5) showed elevated transcript levels in ZR (P[lt]0.01). Among the other transcripts, ZF had increased expression of PCP4 (purkinje cell protein 4), LEF1 (lymphoid enhancer-binding factor 1), PTGDS (prostaglandin D2 synthase) and NOV (nephroblastoma overexpressed gene) while ZR showed increased expression of FGG (fibrinogen gamma chain), SLC27A2 (fatty acid transporter, member 2) and TSPAN12 (tetraspanin 12). [bold]Conclusion:[/bold] Microarray is a technique suitable for the comprehensive analysis of gene expression profiles across different tissues. Using this approach, we have identified several novel candidate genes for elucidating the molecular mechanisms governing the ZF and ZR, thereby increasing our understanding of the functional zonation of these two adrenocortical zones.[br][br]Nothing to Disclose: JR, TW, TDM, WER 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2207 316 2451 MON-577 PO16-01 Monday 2120 2012


2116 ENDO12L_MON-578 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Vitamin D Status and Genotype in Autoimmune Addison Disease from Five European Cohorts Klaus Badenhoop, Gesine Meyer, Heinrich Kahles, Anette B Wolff, Beate Skinningsrud, Corrado Betterle, Alberto Falorni, William Ollier, Anna Kasperlik-Zaluska, Dag Undlien, Eystein Husebye, Simon HS Pearce, Anna L Mitchell, Vincenzo Cerundolo, Marissa Penna-Martinez University Hospital Frankfurt am Main, Frankfurt am Main, Germany; Haukeland University Hospital, Bergen, Norway; Oslo University Hospital, Oslo, Norway; University of Padua, Padua, Italy; University of Perugia, Perugia, Italy; University of Manchester, Manchester, UK; Bielanski Hospital, Warsaw, Poland; Newcastle University, Newcastle, UK; University of Oxford, Oxford, UK [bold]Background: [/bold]Vitamin D deficiency contributes to autoimmune disease and significant associations with the genes controlling its activation and metabolisation have been reported: amongst others in type 1 diabetes, multiple sclerosis and autoimmune Addison[acute]s disease (AAD). Besides its role in bone mineralisation vitamin D is an important regulator of T-lymphocyte and macrophage interaction thus involved in the early steps of immune mediated endocrine organ destruction. In order to define the vitamin D status and its genetics we screened patients with Addison[acute]s disease from five European countries and correlated the findings with genotypes for the vitamin D receptor (VDR), the cholesterol synthesis enzyme 7-dihydrocholesterol reductase (DHCR7), the degrading enzyme 24-hydroxylase CYP24A1 and the activating enzyme 1-a-hydroxylase CYP27B1 using a tag-SNP approach. [bold]Methods: [/bold]One thousand forty-eight AAD cases (Germany n = 237; Italy n = 349; Norway n = 378, UK n = 45, and n=39 Poland) were studied for vitamin D system genotypes and a subgroup of these for the vitamin D status (n=712) using a RIA (DIASORIN) for 25(OH)D[sub]3[/sub]. Taqman genotyping was performed in addition for CYP27B1 (rs10877012), VDR ApaI (rs7975232) and DHCR7 (rs12785878) polymorphisms for samples from Germany (330 cases, 292 controls), and Norway (358 cases, 362 controls). [bold]Results: [/bold]Overall 54-61% patients were vitamin D insufficient or severely (9-17%) deficient ([lt]20 or [lt]10ng/ml), the median in all European cohorts was [lt]20 ng/ml and only 8-13% of patient reached sufficient 25(OH)D[sub]3[/sub] levels [gt] 30ng/ml. The 25(OH)D[sub]3 [/sub]was significantly lower in CYP27B1 (rs10877012) CC homozygotes compared to heterozygotes in German patients and carriers of the allele [ldquo]C[rdquo] had significantly higher thyroid peroxidase antibody (TPO-Ab) titers. DHCR7 (rs12785878) GG homozygotes were significantly more frequent in Norwegian patients in comparison to matched controls (p=0.02). Vitamin D deficient ([lt]20ng/ml) patients showed significantly less frequent alleles of CYP24A1 (p=0.01) and VDR (p[lt]0.05) in comparison to controls, whereas the group with vitamin D levels [gt] 20ng/ml showed no significant differences for these genotypes but for others. [bold]Conclusion:[/bold] These data suggest a contribution of the vitamin D system genotypes to autoimmune Addison[acute]s disease depending on the vitamin D status. Since vitamin D deficiency is highly prevalent a targeted supplementation strategy may be necessary to achieve sufficiency in Addison[acute]s disease.[br][br]Nothing to Disclose: KB, GM, HK, ABW, BS, CB, AF, WO, AK-Z, DU, EH, SHSP, ALM, VC, MP-M 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1914 316 2452 MON-578 PO16-01 Monday 2121 2012


2117 ENDO12L_MON-579 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Cubilin Exon Polymorphism and Its Effects on Vitamin D Levels in Differentiated Thyroid Carcinoma Marissa Penna-Martinez, Flavia Angeletti, Heinrich Kahles, Klaus Badenhoop University Hospital Frankfurt am Main, Frankfurt am Main, Germany [bold]Background:[/bold] Cubilin is a multiligand endocytic receptor located in various epithelial cells including the human thyroid gland. It mediates the vitamin D uptake into the cells by binding vitamin D metabolites complexed with the vitamin D binding protein (DBP). A relation between lower levels of circulating 1,25(OH)[sub]2[/sub]D[sub]3[/sub] and differentiated thyroid cancer (DTC) was reported by our group previously. The aim of the present study was to investigate the role of Cubilin (rs1801222) exon polymorphism in patients with DTC and healthy controls (HC). Also its influence on the vitamin D status in DTC was evaluated. [bold]Method:[/bold] Patients (n = 230; 153 females and 77 males) with DTC (follicular, n = 44 or papillary, n = 186) and HC (n = 294; 130 females and 164 males) were genotyped for the Cubilin polymorphism (rs1801222) using real time PCR. Furthermore, the 25(OH)D[sub]3[/sub], and 1,25(OH)[sub]2[/sub]D[sub]3[/sub] plasma levels in patients were measured by radioimmunoassay. [bold]Results[/bold]: No difference was observed between DTC patients and the HC in the genotype frequencies. However when patients were devided into papillary (PTC) and follicular thyroid cancer patients and grouped according to four 25(OH)D[sub]3[/sub] categories (severely deficient, deficient, insufficient, sufficient), in the deficient PTC group the allele G was more prevalent (75% vs. 54.9%), while the allele A was found to be less prevalent (25% vs. 45.1%; p = 0.004) compared to the HC. Furthermore, the genotype analysis revealed that the genotype GG was more frequent (58.3% vs. 29.3%), whereas the genotypes AG (33.3% vs. 51.2%) as well as AA (8.3% vs. 19.5%; p = 0.01) were less frequent than in the HC. Additionally, deficient PTC patients with genotype GG showed significantly lower 1,25(OH)[sub]2[/sub]D[sub]3[/sub] plasma levels compared with the HC (p = 0.0004). [bold]Conclusion:[/bold] A combination of the allele G of the Cubilin polymorphism (rs1801222) and a deficient vitamin D status with lower 1,25(OH)[sub]2[/sub]D[sub]3[/sub] activation appear to be associated with an increased risk for PTC.[br][br]Nothing to Disclose: MP-M, FA, HK, KB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1760 316 2453 MON-579 PO16-01 Monday 2122 2012


2118 ENDO12L_MON-580 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Combined Inactivation of [italic]Foxd3[/italic] and [italic]Sdhb[/italic], in Mice, Results in Postnatal Lethality Eva Szarek, Allessio Giubelino, Karel Pacak, Anelia Horvath, Matthew F Starost, James L Weed, Maria Nesterova, Constantine A Stratakis Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, MD; Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, MD; National Institutes of Health (NIH), Bethesda, MD [italic]FOXD3/Foxd3[/italic] and [italic]SDHB/Sdhb[/italic] are tumor suppressor genes. [italic]FOXD3/Foxd3[/italic] (human locus 1p31.3) is required for maintenance of neural crest progenitors in the embryo, whereas [italic]SDHB/Sdhb[/italic] (human locus 1p35-36.1) is one of four subunits making up succinate dehydrogenase that play essential roles in the tricarboxylic acid cycle and aerobic respiratory chain. Mutations in [italic]SDHB[/italic], as well as [italic]SDHC[/italic] and [italic]SDHD[/italic], have been associated with Carney-Stratakis syndrome [CSS; OMIM#606864], an autosomal dominant syndrome characterized by paraganglioma (PGL) and gastrointestinal stromal tumors (GISTs) or the non-familial Carney Triad [CT; OMIM#604287], a condition associated with PGLs and GISTs with pulmonary chondromas and other tumors; affecting mostly pediatric and young adult patients. Recently, our group identified 3 [italic]FOXD3[/italic] coding variants in 3 unrelated patients with sporadic non-[italic]KIT/PDGFRA[/italic]-GISTs among 24 patients. To determine the consequence of mutations in both genes, defects in [italic]Foxd3[/italic] and [italic]Sdhb[/italic] were combined in one mouse model. [italic]Foxd3[sup]-/-[/sup][/italic] mouse embryos die after implantation at approximately 6.5 days (d), therefore we used [italic]Foxd3[sup]+/-[/sup][/italic] mice for mating with [italic]Sdhb[sup]+/-[/sup][/italic] mice. Interestingly, at weaning (21-28d) the Mendelian frequency was: 40% wild-type (wt); 40% [italic]Foxd3[sup]+/-[/sup][/italic].wt; 20% [italic]Sdhb[sup]+/-[/sup][/italic].wt; 0% [italic]Foxd3[sup]+/-[/sup][/italic].[italic]Sdhb[sup]+/-[/sup][/italic]. Thus, [italic]Foxd3/Sdhb[/italic] double-mutant mice were not detected postnatally. To determine whether the [italic]Foxd3/Sdhb[/italic] double mutation was embryonically lethal we examined embryos from embryonic day (E) 10.5 until birth. During embryonic development a normal Mendelian frequency (16% wt; 24% [italic]Foxd3[sup]+/-[/sup][/italic].wt; 16% [italic]Sdhb[sup]+/-[/sup][/italic].wt; 44% [italic]Foxd3[sup]+/-[/sup][/italic].[italic]Sdhb[sup]+/-[/sup][/italic]) was observed. Given the role of [italic]Foxd3[/italic] and [italic]Sdhb[/italic] during development we hypothesized that the [italic]Foxd3/Sdhb[/italic] double mutation leads to death for newborn pups. Interestingly, one male pup from one [italic]Foxd3[sup]+/-[/sup][/italic] x [italic]Sdhb[sup]+/- [/sup][/italic]mating survived, until 20d postnatal. The animal weighed 2.44g and measured 3.5cm. Upon opening the carcass, scant adipose stores were observed and there was scant to no ingesta/digesta throughout the gastrointestinal tract. We conclude that combined heterozygosity for [italic]Sdhb[/italic] and [italic]Foxd3[/italic] leads to early postnatal lethality in mice of a mixed genetic background. Both genes are on chromosome 1p, a loci frequently lost in tumors associated with Carney Triad.[br][br]Nothing to Disclose: ES, AG, KP, AH, MFS, JLW, MN, CAS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2003 316 2454 MON-580 PO16-01 Monday 2123 2012


2119 ENDO12L_MON-581 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) [italic]Galgeun-tang[/italic] Inhibits Atopic Dermatitis Responses in Dust Mite Extract-Treated Nc/Nga Mice Hyekyung Ha, Hoyoung Lee, Hye-Sun Lim, Mee-Young Lee, Chang-Seob Seo, Hyeunkyoo Shin Korea Institute of Oriental Medicine, Daejeon, Korea [italic]Galgeun-tang[/italic] (GGT, [italic]gegen[/italic] decoction, [italic]kakkon-to[/italic]), one of common herbal formulas, includes seven herbs such as Puerariae Radix (major component), Cinnamomi Cortex Spissus, Ephedra Herba, Paeoniae Radix, Glycyrrhizae Radix et Rhizoma, Zingiberis Rhizoma Crudus and Zizyphi Fructus. GGT is used to treat common cold, fever, headaches, hangover and neck/upper back stiffness.[br]This study was to confirm the efficacy of GGT for treating atopic dermatitis (AD) in house dust mite (Biostir-AD[sup][reg][/sup]) applied Nc/Nga mice. We measured the thymus- and activation-regulated chemokine (TARC) level on human keratinocyte cell line (HaCaT). The HaCaT had been stimulated with TNF-[alpha] (10 ng/ml) and IFN-[gamma] (10 ng/ml). The male Nc/Nga mice were induced AD by Biostir-AD. GGT was given to mice orally at doses of 300 mg/kg/day, p.o. during 6 weeks. GGT has inhibitory effect of TARC production in TNF-[alpha]/IFN-[gamma] stimulated HaCaT cells, dose-dependent manner (p[lt]0.01). In Biostir-AD applied Nc/Nga mice, the plasma levels of total IgE and histamine were significantly increased in the control group compared with that to the normal group (p[lt]0.01). GGT were inhibited increasing the plasma IgE and histamine levels in Biostir-AD sensitized mice (p[lt]0.01). Repeated administration of GGT inhibited of development of AD in Nc/Nga mice. These beneficial effects may at least in part be associated with the inhibitory effect of GGT on IgE and histamine production in dermatitis lesions. The GGT will be suggested a candidate for the treatment of AD.[br][br]Sources of Research Support: This research was supported by a grant from Korea Institute of Oriental Medicine, #K12030.[br][br]Nothing to Disclose: HH, HL, H-SL, M-YL, C-SS, HS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1462 316 2455 MON-581 PO16-01 Monday 2124 2012


2120 ENDO12L_MON-582 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Molecular Determinant of the Diversity of Thyroid Hormones and Their Receptors[apos] Action Nicolas Buisine, Alexis Grimaldi, Gladys Alfama, Barbara A Demeneix, Xiaoan Ruan, Yijun Ruan, Laurent M Sachs Museum National d[apos]Histoire Naturelle, Paris, France; Agency for Science, Technology and Research, Singapore, Singapore Thyroid hormones (THs) are versatile players. They regulate diverse cellular processes from mitosis to apoptosis, from metabolism to growth and development. Specific regulation provides a challenge for researchers and physicians. One striking developmental processes orchestrated by THs, is amphibian metamorphosis. Tadpole transformation is marked by dramatic changes including de novo morphogenesis (limb), tissue remodelling (brain, intestine[hellip]) and organ resorption through apoptosis (tail). These changes involve cascades of gene regulation initiated by THs and their receptors (TR). Moreover, metamorphosis has close and interesting parallels with the perinatal period in mammals and is thus an attractive model. Indeed, both periods coincide with a peak of THs that have roles, notably in maturation of the nervous system and the intestine. Our aim was to build a genome wide map of TH direct and indirect responsive genes and then using the function associated to these genes to highlight the difference between TH programs leading to different cell fates. Transcriptome analysis were carried out in two tissues showing opposite cell fate during metamorphosis: the limb and the tail fin. THs response genes mapping was derived from RNA-Seq analysis. Direct response was sorted out by linking the transcription start sites of target genes using RNA-PET analysis with TR binding sites using chromatin interaction analysis by paired-end tag sequencing (ChIA-PET). Indeed, ChIA-PET not only allow to map TR binding sites but also the physical interactions between them and transcription start sites of regulated genes. The results open new perspective to understand gene regulations controlled by TH and the potential molecular determinants involved in the diversity of TH effects. Finally, the characterisation of such regulatory programs will help understand how the dysfunction of THs signal can have deleterious effects on human health. Thus, the extraction of pertinent results will be carried out in the light of the impact of THs on human health and quality of life.[br][br]Sources of Research Support: CNRS, MNHN, A-Star, EU FP6 Crescendo, EU FP7 Ideal, ANR JCJC Trigger, PHC Merlion.[br][br]Nothing to Disclose: NB, AG, GA, BAD, XR, YR, LMS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1196 316 2456 MON-582 PO16-01 Monday 2125 2012


2121 ENDO12L_MON-583 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Hypoxia Inducible Factor-3 Is a Transcriptional Activator That Up-Regulates Gene Expression under Hypoxia Peng Zhang, Qing Yao, Ling Lu, Cunming Duan Ocean University of China, Qingdao, China; University of Michigan, Ann Arbor, MI Hypoxia-inducible factors (HIFs) are key transcriptional regulators coordinating the physiological responses to hypoxia. A prevailing view is that while HIF-1 and -2 are transcriptional activators, HIF-3 acts as a negative inhibitor of HIF-1[alpha] and -2[alpha] action. Here we report multiple lines of in vitro and in vivo evidence suggesting that Hif-3 functions as a transcriptional activator and plays an indispensible role in regulating gene expression under hypoxia in zebrafish. Zebrafish hif-3[alpha] mRNA is expressed in many embryonic and adult tissues and is regulated by hypoxia at mRNA and protein levels. Mutational analysis revealed that residue P493 is most critical for its oxygen-dependent degradation, while P393 and L503 also contribute. Two clusters of basic residues in the C-terminal region of Hif-3[alpha] acted redundantly to regulate its nuclear localization. When tested in cultured human cells in vitro and zebrafish embryos in vivo, Hif-3[alpha] had strong activity in activating HRE-dependent reporter gene expression. A long variant of human HIF-3[alpha] had similar activity. Forced expression of Hif-3[alpha] in zebrafish embryos induced the expression of several genes that were distinct from the Hif-1[alpha] target genes. Dominant-negative inhibition and morpholino knockdown of Hif-3[alpha] blunted hypoxia-induced expression of these Hif-3-regulated genes. These results suggest that Hif-3[alpha] is a transcriptional activator and plays an important role in regulating gene expression in zebrafish embryos under hypoxia.[br][br]Sources of Research Support: This study was supported by Natural Scientific Foundation of China Grant# 30970357, 30928021, 30800838, and by US National Science Foundation Research Grant IOS-1051034.[br][br]Nothing to Disclose: PZ, QY, LL, CD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2303 316 2457 MON-583 PO16-01 Monday 2126 2012


2122 ENDO12L_MON-584 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Comprehensive Temporal Expression Profiling Reveals a Complete Natriuretic Peptide System in the Developing Zebrafish ([italic]Danio rerio[/italic]) Andrew J Lessey, Samantha M Mirczuk, Charlotte A Dawson, Imelda M McGonnell, Robert C Fowkes Royal Veterinary College, University of London, London, UK; Royal Veterinary College, University of London, London, UK C-type natriuretic peptide (CNP) is the most highly conserved member of the mammalian natriuretic peptide family. Although broadly expressed throughout the periphery, CNP is the major natriuretic peptide of the central nervous system and is suspected to influence neuroendocrine function. In fish species, CNP also plays an important role in osmoregulation. Our recent studies using human fetal pituitary samples has shown that the gene encoding CNP, [italic]NPPC[/italic], is expressed early on in pituitary development but a comprehensive understanding of CNP expression [italic]in utero[/italic] remains to be established. We have taken advantage of the highly versatile [italic]Danio rerio[/italic] (Zebrafish) as an excellent vertebrate model of development to establish which components of the natriuretic peptide system are expressed throughout development. Zebrafish were marbled twice a week and embryos were captured at time points ranging from 3hpf up to 120hpf, prior to extraction of total RNA, DNase clean-up and cDNA generation. Additionally, adult Zebrafish were euthanized and total RNA was extracted from both head and body separately. RT-PCR enabled us to screen for the expression of [italic]nppa[/italic] (ANP gene), [italic]nppb[/italic], [italic]BNP[/italic] (BNP genes), [italic]nppc2, cnp3, nppc4, nppcl[/italic] (CNP genes), [italic]npr1a, nprA[/italic] (GC-A genes), [italic]nprB[/italic] (GC-B) and [italic]nprC[/italic] (clearance receptor gene). All these genes were expressed, but clear temporal differences between the four distinct CNP transcripts were observed. There was some evidence of enhanced transcript levels at 3hpf, consistent with the presence of residual maternal transcripts prior to the maternal-to-zygote transition. [italic]In silico[/italic] phylogenetic analysis of the four CNP genes demonstrated that evolutionary conservation was maintained between these transcripts in Zebrafish and across other fish species (Lamprey, Japanese Medaka, Fugu and Rainbow trout). These data reveal the presence of an intact natriuretic peptide system in developing Zebrafish, and will facilitate a reverse genetics approach to identify their functional roles within neuroendocrine development.[br][br]Sources of Research Support: Wellcome Trust Grant WT093257MA awarded to RCF [amp] IMM.[br][br]Nothing to Disclose: AJL, SMM, CAD, IMM, RCF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1515 316 2458 MON-584 PO16-01 Monday 2127 2012


2123 ENDO12L_MON-585 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) Functional Characterization of a Novel Mutation (Ivs9+5g[gt]A) in the Human Aromatase Gene Associated with Aromatase Deficiency Nora Saraco, Romina Sainz, Suzana Nesi-Franca, Roxana Marino, Rosana Marques-Pereira, Julia LaPastina, Natalia Perez-Garrido, Romulo Sandrini, Marco A Rivarola, Luiz De Lacerda, Alicia Belgorosky Hospital de Pediatria Garrahan, Buenos Aires, Argentina; Federal University of Parana, Curitiba, Brazil Aromatase (P450arom) is the key enzyme for estrogen biosynthesis and, in humans, is encoded by the CYP19A1 gene. Several mutations in the CYP19A1 gene have been associated with aromatase deficiency. We have previously reported a novel CYP19A1 intronic homozygote mutation (IVS9+5G[gt]A) in an affected 46,XX DSD girl born of non-consanguineous parents presenting an aromatase deficient phenotype characterized by ambiguous genitalia, delayed bone age, absence of breast development, multicystic enlarged ovaries (7 and 10 cc, right and left, respectively) and high serum LH and FSH levels (1). Analysis of the splicing donor site in intron 9 by several splicing prediction programs predicts the disappearance of this site in the presence of the mutation. The aim of this study was to functionally characterize the IVS9+5G[gt]A mutation.[br]P450arom mRNA was analyzed in the patient[apos]s lymphocytes (PBL) by reverse transcription polymerase chain reaction (RT-PCR). Splicing assays were performed in Y1 cells transfected with wild type (WT) or mutant (Mut) minigen constructions, which contain full length exon 9 and each flanking intronic sequences. Protein analysis was carried out by western blot after the transfection of an expression vector containing P450arom cDNA including part of intron 9 (IN9cDNA); a complete wild type P450arom cDNA expression vector (WTcDNA) was used as normal control. Enzyme activity was evaluated by measurement of estradiol production after transfection of IN9cDNA or WTcDNA into COS cells, using testosterone as substrate.[br]Patient PBL P450arom mRNA analysis showed the inclusion of intron 9, confirmed by sequencing. WT minigene expression resulted in a normally spliced mRNA whereas the mutant minigene generated an abnormal mRNA. Protein analysis showed the expression of a shorter and inactive protein compared with the wild type.[br]In summary, we report a novel IVS9+5G[gt]A mutation in the CYP19A1 gene that generates an mRNA that includes intron 9. The presence of an in-frame stop codon (TAA) 18 bp downstream the splice junction in intron 9 would result in the synthesis of a truncated and inactive aromatase protein lacking the heme-binding region. These findings explain the aromatase deficiency clinical phenotype found in the affected girl.[br][br]1[P1-323] Aromatase Deficiency in a 17-Year-Old Brazilian Girl: Case Report and Description of a Novel Mutation (Endo 2011). S Nesi-Franca, R Marino, R Marques-Pereira, J La Pastina, N Perez Garrido, N Saraco, R Sandrini, MA Rivarola, A Belgorosky, L De Lacerda. Federal University of Parana, Curitiba, Brazil; Hospital de Pediatria Garrahan, Buenos Aires, Argentina.[br][br]Sources of Research Support: FONCYT,CONICET, Argentina.[br][br]Nothing to Disclose: NS, RS, SN-F, RM, RM-P, JL, NP-G, RS, MAR, LDL, AB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 280 316 2459 MON-585 PO16-01 Monday 2128 2012


2124 ENDO12L_MON-586 POSTER SESSION: Gene Expression Regulatory Networks in Endocrine Systems (1:30 PM-3:30 PM) The Core Clock Gene, Bmal1, Promotes Skeletal Muscle Development Via Wnt Pathway Somik Chatterjee, Ji M Kim, Zhe Fang, David Nelson, Laurie J Minze, Willa A Hsueh, Ke Ma The Methodist Hospital Research Institute, Houston, TX; Baylor College of Medicine, Houston, TX Circadian clock present in many tissues is an intrinsic regulatory mechanism of metabolism and physiology, and recent studies suggest it may regulate developmental pathways. We hypothesized that the core clock gene, Bmal1, could play a role in muscle development. We found that Bmal1-null mice exhibit significantly reduced muscle mass with smaller muscle fiber. We further demonstrate this is due to a cell-autonomous function of Bmal1 as stable shRNA knockdown (KD) in C2C12 myoblasts markedly suppressed myogenic differentiation with down-regulation of key myogenic factors (Myf5, MyoD, myogenin) and muscle-specific markers, as assessed by RT-qPCR and immunostaining. Primary myoblasts from Bmal1-null mice displayed a similar myogenic defect. In contrast, Bmal1 overexpression promoted myogenesis and increased myogenic factor expression. By chromatin immunoprecipitation, we identified Bmal1 occupancy on promoters of many canonical Wnt signaling components, including Wnt10a, Dishevelled2 and TCF3. This finding indicates that Bmal1 directly regulates Wnt pathway, a known mechanism of promoting skeletal muscle development. Among these genes, Wnt10a was markedly down-regulated in Bmal1-null primary myoblasts, and Wnt3a treatment was able to restore the compromised Wnt signaling activity. Strikingly, in C3H10T1/2 mesenchymal stem cells, suppressed Wnt signaling by attenuation of Bmal1 function severely blocked myogenic lineage while promoted differentiation toward the adipocyte fate, suggesting Bmal1 is involved in mesenchymal lineage allocation. Taken together, our study demonstrates that Bmal1 functions as a key regulator of myogenesis at both lineage determination and terminal differentiation processes, and this is mediated by direct transcriptional regulation of the Wnt signaling pathway.[br][br]Nothing to Disclose: SC, JMK, ZF, DN, LJM, WAH, KM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1024 316 2460 MON-586 PO16-01 Monday 2129 2012


2125 ENDO12L_MON-588 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Pseudohypoparathyroidism Type Ib Presenting with Mild Congenital Hypothyroidism [mdash] A Previously Unreported Association Carolina C Di Blasi, Craig E Taplin Seattle Children[apos]s Hospital and the University of Washington, Seattle, WA [underline][bold]Background:[/bold][/underline] Pseudohypoparathyroidism type Ib (PHP-Ib) is classically characterized by renal resistance to PTH in the absence of other endocrine or physical abnormalities and is a maternally inherited disorder. While TSH resistance has been described in PHP type 1a, reports of hypothyroidism are rare in PHP-Ib. Recently, resistance to the action of TSH has been documented in adults with PHP-Ib (1).[br][bold][underline]Clinical Case:[/underline][/bold] A 13 yo boy with a history of mild infantile hypothyroidism presented with hypocalcemia, hyperphosphatemia and elevated iPTH. His newborn screen TSH obtained on day one of life was 23.89 uIU/ml with normal T4. At 2 months of age he had jaundice and difficulties sucking, with mild persistent elevations in TSH throughout the first year. At 12 months of age, his TSH was 8 uIU/ml (0.5-4.5) and thyroid hormone replacement was commenced. Tc-99 scan demostrated normal uptake in an orthotopic bilobed gland. Thyroid replacement was continued until age 9. Therapy was stopped when he was lost to follow-up.[br]Three years later workup was obtained in the context of limb pain. He had hypocalcemia with Calcium of 6.3 mg/dl (8.7-10.7). There was no history of seizures or tetany. TSH was mildly elevated at 4.85 uIU/ml, while other results showed 25-OH-Vitamin D of 24 ng/ml (30-100) and ALP 370 U/L (119-426). Repeat workup demostrated persistent hypocalcemia (total Calcium 5.8 mg/dl, ionized Calcium 0.81 mmol/L) with hyperphosphatemia (9.2 mg/dl) and elevated iPTH of 376 pg/ml (12-75). Mild thyroid dysfunction persisted with a TSH of 4.73 uIU/ml but normal T4 of 5.6 mcg/dl (4.5-10). He had no other endocrine disturbances identified with normal Testosterone and IGF1 levels. A bone age x-ray showed no evidence of short metacarpals. Height was on the 11th percentile, consistent with his mid-parental height. A maternal history of adquired hypothyroidism and seizures secondary to hypocalcemia requiring Calcitriol was uncovered.[br]He was diagnosed with PHP-Ib based on the hypocalcemia, hyperphosphatemia and high iPTH in the absence of stigmata of Albright[apos]s hereditary osteodystrophy. His TSH remains above the upper normal reference limit. He was started on Calcitriol and Calcium.[br][underline][bold]Conclusion:[/bold][/underline] PHP-Ib has not been previously described in association with infantile hypothyroidism. We conclude that where the pathophysiology of mild congenital hypothyroidism is unclear, follow-up for evidence of PHP is indicated.[br][br](1) Mantovani et al.,J Clin Endocrinol Metab. 2007 Sep; 92(9): 3738-42.[br][br]Nothing to Disclose: CCDB, CET 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 277 317 2461 MON-588 PO54-02 Monday 2130 2012


2126 ENDO12L_MON-589 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Vitamin D Intoxication in a Toddler Due to a Dispensing Error of an Imported Vitamin D Supplement Larisa Rusyn, Payal R Patel, Raphael David, Brenda Kohn, Preneet C Brar New York University School of Medicine, New York, NY [bold]Introduction[/bold]: Vitamin D intoxication is a rare cause of hypercalcemia. There is a recent increase in reports of pediatric cases of hypercalcemia, secondary to vitamin D intoxication. This trend is attributable to the growing popularity of vitamin D supplements, especially as Vitamin D is being marketed as a [quot]panacea[rdquo] for a number of medical problems.[br]Immigrant families often travel to their countries of origin, where vitamin supplements are available in formulations different from the US. Parents may not report using supplements to their child[apos]s pediatrician, as these supplements are perceived [quot]safe[quot] with no dangerous side effects.[br][bold]Case Presentation[/bold]: A three year old toddler presented to the emergency room with emesis, polyuria, and lethargy. On inquiry, the family reported use of a multivitamin gel (made in Ecuador) dispensed at 1 tsp bid. Per the manufacturer[apos]s label, 5 grams ([asymp]one teaspoon) contains 0.096 mg ([asymp]100 IU) of cholecalciferol. A standard conversion of 1[micro]g of cholecalciferol equals 40 IU[sup]1[/sup], secondary to conversion error this patient received 7,680 IU per day and 108,000 IU total vitamin D3 over 14 days, which is toxic. [quot]Teaspoon[quot] dosage inaccuracies and potentially excessive administration by caretakers compounded vitamin D toxicity.[br]Initial lab values revealed Ca 16 mg/dl, iPTH 2.25 pg/mL (15-65), 25 OH vit D3: [gt]512ng/mL(30-100), and UCa/UCreatinine 1.14mg/mg([lt]0.2). Hydration, lasix, calcitonin (6 days), and prednisolone (7 days) were used to manage the hypercalcemia. After 15 days of treatment, the 25 OH vitamin D3 was 324 ng/ml and the calcium decreased to 11.2 mg/dL.[br][bold]Conclusions[/bold]: In our case, dispensing and labeling errors led the parents to administer high doses of vitamin D to their toddler, leading to hypercalcemia. This highlights several issues regarding vitamin supplement usage.[br]First, physicians should be aware of the dosage conversion of vitamin D in terms of IU, mg and mcg to calculate if the prescribed dose is appropriate for their patients. In addition, as there are no established manufacturing standards for certain vitamin supplement formulations[mdash]such as gels[mdash]caution should be exercised as more reports are emerging of toxic ingestion from overzealous supplementation. Verification of supplement source and composition is essential. Lastly, educating parents about the safe use of vitamin D supplements[mdash]emphasizing the need to report imported supplements to the patient[apos]s pediatrician[mdash]is important to avoiding erroneous toxic ingestion.[br][br](1)Ryan B Jacobsen et al., Hypervitaminosis D Associated with a Vitamin D Dispensing Error, The Annals of Pharmacotherapy 2011; 45:e52.[br][br]Nothing to Disclose: LR, PRP, RD, BK, PCB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 704 317 2462 MON-589 PO54-02 Monday 2131 2012


2127 ENDO12L_MON-590 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Selenoprotein Deficiency Syndrome Caused by Novel Heterozygous Mutations in the SBP2 Gene Laura C Sillers, Nazli Gonc, Xiao-Hui Liao, Nugun Kandemir, Ayfer Alikasifoglu, Alexandra M Dumitrescu, Roy E Weiss, Samuel Refetoff University of Chicago, Chicago, IL; University of Chicago, Chicago, IL; Hacettepe University, Ankara, Turkey Selenoproteins have an essential role in protection from oxidative stress and in thyroid hormone metabolism, among other functions. The rare amino acid selenocysteine (Sec) is incorporated into these peptides through a highly regulated process, with Sec insertion sequence-binding protein 2 (SBP2) being one of the limiting factors. Fewer than ten affected individuals with SBP2 mutations have been reported worldwide (1). These patients have abnormal thyroid function tests (TFTs) and variable phenotypes of impaired growth, motor and mental delay, myopathy, and photosensitivity.[br]An 11 year-old Turkish girl born to non-consanguineous parents presented with mental (IQ of 50) and motor retardation, poor growth (height and weight [le] 10th percentile), and abnormal TFTs including low T3 (50ng/dl; normal 90-200), high T4 (14.4[micro]g/dl; 5-12) and freeT4 Index (15.4; 6-10.6), very high reverseT3 (113ng/dl; 16-34), and mildly elevated TSH (3.7mU/L; 0.3-3.6), a phenotype similar to that of patients with SBP2 deficiency. DNA sequencing of the SBP2 gene revealed compound heterozygous mutations [ndash] a 4bp deletion in exon 14 inherited from the father (c.2045-2048 del resulting in p.L681 frame shift 683X) and a nonsense mutation in exon 16 (c.2344C-[gt]T p.Q782X), inherited from the mother. Genotyping of the family showed the patient[apos]s younger brother was carrier for the maternal mutant SBP2, while the older sister had normal SBP2 sequence.[br]The SBP2 gene has 17 exons, with critical functional domains located in the carboxyterminus encoded by exons 11 through 16. There are smaller isoforms generated from alternative splicing of early exons encoding the aminoterminus, which are believed to account for the relatively mild phenotype observed in individuals with early nonsense mutations. Though both of this patient[apos]s alleles are mutated in the highly functional C-terminus, we believe that the Q782X protein is truncated shortly after the region necessary for SBP2 activity, as suggested by in vitro studies using the rat Sbp2 protein (2), thus preventing a more severe or even lethal phenotype.[br]Conclusion: SBP2 deficiency is a rare condition in which there is impaired synthesis of selenoproteins. The thyroid phenotype of high rT3, high total and free T4, low T3, and normal to slightly increased TSH seems to be a consistent finding. Identification of additional patients bearing this deficiency may improve our understanding of this syndrome and the role of selenoproteins in human physiology.[br][br](1) Dumitrescu, AM, Refetoff, S., Ann Endocrinol 2011;72:95-98. (2) Bubenik JL and Driscoll DM. J Bio Chem 2007;282:34653-62.[br][br]Sources of Research Support: NIH Grant DK15070 awarded to SR, DK091016 awarded to AMD, and DK064582 awarded to LCS.[br][br]Disclosures: SR: Consultant, Quest Diagnostics. Nothing to Disclose: LCS, NG, X-HL, NK, AA, AMD, REW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2149 317 2463 MON-590 PO54-02 Monday 2132 2012


2128 ENDO12L_MON-591 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Hyperthyroidism in McCune-Albright Syndrome with Incalcitrant Tachycardia Alicia M Diaz-Thomas University of Tennessee Heath Science Center, Memphis, TN Background: McCune Albright syndrome (MAS) is caused by a post zygotic somatic gain-of-function mutation in the GNAS1 gene that is distributed in a mosaic fashion in various tissues of the affected individual. The classic triad of MAS includes polyostotic fibrous dysplasia, caf[eacute]-au-lait skin pigmentation and precocious puberty. Thyroid involvement is common and ranges from frank hyperthyroidism to structural abnormalities of the thyroid gland itself.[br]Clinical case: A 16y 8mo old male with MAS was referred to the pediatric endocrine clinic for evaluation of abnormal thyroid function tests discovered during intake assessment to a psychiatric hospital for suicidal ideation. There he was found to be hypertensive (systolic BP of 130[apos]s-140[apos]s) and tachycardic (HR of 120[apos]s 130[apos]s). Hypothyrotropinemia (TSH 0.013 mIU/mL RR: 0.5-4.50) was noted with a low normal thyroxine level (TT4 4.9 ug/dL RR: 4.5-12.0). He was started on Lopressor 20 mg BID which was increased to 50 mg BID. Subsequent evaluation revealed a depressed, dysmorphic young man in a wheelchair. He was tachycardic with a hyperdynamic precordium and had tremor of the hands and tongue fasciculation. Further evaluation of thyroid function revealed: TSH [lt]0.005 mIU/mL RR: 0.45-4.50, FT4 1.31 ng/dL RR: 0.93-1.6, TT3 180ng/dL RR: 71-180, FT3 4.8pg/mL RR: 2.3-5.0, rT3 403 pg/mL RR: 90-350, TSII 58% R: 0-139, TRAb [lt]0.51IU/L RR: 0.00-1.75, and undetectable thyroid peroxidase and thyroglobulin antibodies. I [sup]123 [/sup]thyroid uptake and scan revealed a normal sized thyroid gland without nodules and an increased uptake (38% RR: 8-35%) at 24 hours. Radioablation was performed several weeks later with 25.6miCu of I [sup]131[/sup]. Thyroxine supplementation was started 4 months after radioablation when the patient developed hypothyroxinemia (TSH 2.620 mIU/mL RR: 0.45-4.50, FT4 0.54 ng/dL RR: 0.93-1.6) He continued to receive lopressor for tachycardia and was referred for echocardiogram. Suicidal ideation had resolved.[br]Conclusion: This patient had overt signs and symptoms of hyperthyroidism and hypothyrotropinemia, but lacking a measureable increase in thyroid hormones or an increase in the T3/T4 ratio. His initial presentation of suicidal ideation has resolved, but his tachycardia remains. Insofar as G-stimulated proteins are involved in the signaling cascade of cardiac muscle, perhaps this patient[apos]s cardiac tissue also harbors the GNAS1 mutation which may be responsible for the inotropic effect.[br][br]Nothing to Disclose: AMD-T 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1827 317 2464 MON-591 PO54-02 Monday 2133 2012


2129 ENDO12L_MON-592 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Long-Term Follow-Up of a Child with Recurrent Hyperthyroidism in the Absence of TSH Receptor Antibodies Christopher John Dunne, Francesco De Luca St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA [bold]Background[/bold]: Few patients with autoimmune thyroiditis initially experience biochemical hyperthyroidism in the absence of TSH receptor antibodies. This phase typically lasts only a few weeks, and it is followed by permanent euthyroidism or hypothyroidism. We report a child with recurrent hyperthyroidism not due to Graves[apos] disease.[br][bold]Clinical case[/bold]: E.H. was a 4 and 6/12 year old male who was referred to Endocrinology because of a high TSH. Upon evaluation, the TSH was 9.44 mIU/L (nl. 0.70-6.40), the free T4 1.1 ng/dL (nl. 0.9-1.6), the thyroglobulin (TG) antibody titer was high (85 IU/mL, nl. [lt]20) and the TPO antibody titer was normal ([lt]10 IU/mL, nl.[lt]35). The thyroid size was normal and his review of systems revealed constipation, speech delay, and ADHD. Based on these findings, L-thyroxine (50 mg) replacement was initiated.[br]1 year later, his mother admitted to not being adherent to the L-thyroxine treatment. Due to his normal free T4 and TSH (1.05 ng/dl and 4.49 mIU/dL respectively), she was advised to discontinue the treatment. During the subsequent year, he was repeatedly found to have normal free T4 and TSH.[br]At age7, E.H. was referred again to us due to increased hyperactivity and sleep difficulties. At that time, he was found with suppressed TSH ([lt]0.01 mIU/L), high free T4 (2.8 ng/dL) high TG titer (312 IU/mL) and normal TSI (90%, normal [lt]125%). A [sup]123[/sup]I uptake was borderline-low (4 hrs.: 7.3%, nl.l 5-15%. 24 hrs: 1.7%, nl. 10-40%).[br]6 wks later, repeat laboratory studies revealed normal free T4 and total T3 (1.2ng/dL and 128ng/dL, respectively) and borderline-high TSH (5.49 mIU/L). 6 months later, E.H. was clinically and biochemically euthyroid (TSH 3.34mIU/L and free T4 1.0ng/dL), and he was found repeatedly euthyroid in the following 22 months.[br]At 9 yr 10/12, E.H. was noted by his mother to be more hyperactive, and with increased appetite and thirst. Upon evaluation, he was found with a persistently normal thyroid size. The TSH was low (0.02 mIU/L) and the free T4 high (2.1 ng/dL). In addition, the TG titer was high (185 IU/mL), the TPO normal, while the TSI was negative (27%).[br][bold]Conclusions[/bold]: To our knowledge, this is the first reported case of a child with autoimmune thyroiditis and recurrent hyperthyroidism not due to Graves[apos] disease. The unpredictable course of this child[apos]s thyroid disorder suggests the need for a long-term follow-up in any child with autoimmune thyroid disease.[br][br]Nothing to Disclose: CJD, FDL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 960 317 2465 MON-592 PO54-02 Monday 2134 2012


2130 ENDO12L_MON-593 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Congenital Hypothyroidism Caused by Excess Prenatal Maternal Iodine Ingestion: A Case Series Kara Connelly, Bruce Boston, Elizabeth Pearce, David Sesser, Sam Pino, Lewis Braverman, David Snyder, Stephen LaFranchi Oregon Health [amp] Science University, Portland, OR; Boston University School of Medicine, Boston, MA; State of Oregon Public Health Laboratory, Portland, OR; Legacy Emanuel Children[apos]s Hospital Diabetes and Endocrine Center, Portland, OR Introduction[br]Congenital hypothyroidism (CH), detected by newborn screening (NBS) programs, is most commonly caused by thyroid dysgenesis or dyshormonogenesis. The recently reported increase in incidence suggests that other etiologies may be emerging, including milder and transient causes. We report 3 newborns detected with CH, with evidence supporting excess maternal iodine ingestion as the etiology. Novel testing included measurement of serum iodine extracted from NBS filter paper specimens.[br]Clinical Cases:[br]Case 1 is a term infant with CH; 1st NBS normal; 2nd NBS T4 = 4.29 [mu]g/dL (n 7.2-15.7 [mu]g/dL), TSH = 102.8 mU/L (n 1.7-9.1 mU/L). CH confirmed with serum free T4 = 0.47 [mu]g/dL (n 0.9-2.3 [mu]g/dL) and TSH [gt]100 (n 1.7-9.1 mU/L). The infant[apos]s mother took Iodoral tablets containing 12.5 mg of iodine daily throughout pregnancy. Infant[apos]s urine iodine was normal (70 [mu]g/dL, n 42-350 [mu]g/L) after mother discontinued supplemental iodine, but breast milk iodine was elevated (3,228 [micro]g/L, n 5-180 [mu]g/L).[br]Cases 2 and 3 are twins whose mother took Iodoral 12.5 mg daily throughout pregnancy. CH was detected on initial NBS (case 2: T4 = 5.11 [mu]g/dL [n 7.2-15.7 [mu]g/dL], TSH [gt]200 mU/L [n 1.7-9.1 mU/L]; case 3: T4 = 4.64 [n 7.2-15.7], TSH [gt]200 mU/L [n 1.7-9.1 mU/L]) and confirmed by serum sample (case 2: free T4 = 0.5 [mu]g/dL [n 0.9-2.3 [mu]g/dL], TSH 420 [n 1.7-9.1 mU/L]; case 3: free T4 = QNS, TSH 217 [n 1.7-9.1 mU/L]). Mother had elevated serum and urine iodine levels. These infants also had elevated urinary iodine (case 2: 10,474 [mu]g/L; case 3: 693 [mu]g/L, n 42-350 [mu]g/L).[br]Neonatal iodine exposure was confirmed in all three cases by measuring iodine in serum isolated from NBS filter paper samples. Serum iodine levels in the three newborns were 10 times above the mean level of 10 control samples. In cases 2 and 3, levothyroxine was discontinued after excess maternal iodine ingestion was discovered. Thyroid function tests two and four weeks later were normal; the hypothyroidism appears to have been transient in these two cases. Case 1 remains on levothyroxine at 30 months of age.[br]Conclusion[br]These three cases call attention to a potential increase in the use of nutritional supplements containing iodine in amounts 50 fold higher than the RDA during pregnancy. This etiology of CH may be missed if not investigated by history and confirmed by laboratory testing. Excess maternal iodine may be part of the explanation for the reported increasing incidence of CH.[br][br]Nothing to Disclose: KC, BB, EP, DS, SP, LB, DS, SL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1777 317 2466 MON-593 PO54-02 Monday 2135 2012


2131 ENDO12L_MON-594 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Liothyronine Plus Levothyroxine in Treatment of Refractory Autoimmune Thyroiditis Audrey D Briscoe, Michael E Gottschalk University of California San Diego, San Diego, CA; Rady Children[apos]s Hospital San Diego, San Diego, CA Autoimmune thyroiditis is the most common cause of hypothyroidism in areas of iodine sufficiency. It is successfully treated with Levothyroxine (LT4) in most cases. When LT4 doses exceed the theoretical daily requirement, other factors to consider are poor patient compliance, drug interference and malabsorption. Treatment options include: high dose LT4 or changing administration time. This case describes treatment with the addition of Liothyronine (LT3).[br]An 11yr 11mo old Hispanic female presented with symptoms of lethargy, cold intolerance, hair loss, decreased appetite, weight gain, poor growth and physical exam findings of hypothyroidism. Diagnostic labs revealed: TSH 933[mu]IU/ml(0.35-5), T4 1.2[mu]g/dL(4.3-12.5), TPO antibody [gt]70IU/mL(NL[lt]2), Thyroglobulin antibody 1:25,600(NL[lt]1:100). CPK, aldolase, LDH, AST, ALT and GGT were elevated at diagnosis. She began LT4 75mcg daily and her symptoms resolved, TSH normalized and the enzyme levels improved. She was subsequently diagnosed with autoimmune pancytopenia and hepatitis, cirrhosis, SLE, lupus nephritis, glucocorticoid induced diabetes, splenomegaly and esophageal varices. For the first two years she was euthyroid, but over the next 3 years her TSH fluctuated from 1.93-798[mu]IU/mL with good medication compliance. TSH did not improve after changing the timing of her MVI+Fe or switching from generic LT4 to Levoxyl. TSH improved for a short period with BID dosing of LT4. Even on a LT4 dose of 400mcg daily TSH remained elevated. A brain MRI revealed pituitary gland enlargement from chronic TSH elevation. There was concern for atrophic gastritis since she also had difficulty maintaining normal Tacrolimus levels, however multiple endoscopies were normal as were Vitamin B12 values. One week after adding LT3 25mcg daily, TSH decreased from 454.76 to 1.82[mu]IU/mL. Follow up imaging 2.5mo after LT3 showed a decrease in the pituitary size. LT3 therapy was discontinued after 6 months and she is presently euthyroid requiring 125mcg LT4 daily.[br]After oral administration, 70-80% of LT4 is absorbed in the small intestine and can occur inconsistently, whereas 85-95% of LT3 is absorbed and occurs more reliably. Thus, patients with LT4 malabsorption and unexplained high dose requirements may benefit from the addition of LT3 to improve thyroid function.[br][br]Nothing to Disclose: ADB, MEG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 419 317 2467 MON-594 PO54-02 Monday 2136 2012


2132 ENDO12L_MON-595 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Recurrent Lipoatrophic Panniculitis in a Child with Autoimmune Hypothyroidism Zohreh Shoar, Svetlana Lvovich, Harold G Marks, Wellington J Davis III, Francesco De Luca, Jean-Pierre De Chadarevian, Elizabeth A Suarez St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA; St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA; St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA; St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA; St Christopher[apos]s Hospital for Children, Drexel University College of Medicine, Philadelphia, PA Background: Lipoatrophic Panniculitis (LP) is a rare disorder characterized by inflammation and subsequent loss of adipose tissue. An association between LP and autoimmune diseases has been reported; however, the pathogenesis of fat loss remains poorly understood. Here we report a child with recurrent LP and autoimmune hypothyroidism.[br]Clinical Case: An 11-yr old female was admitted to the hospital for a 2-week history of erythematous, non-pitting, bilateral bipedal edema associated with intermittent fever. She had high TSH (207 uIU/mL), low free T4 (0.6 ng/dL), detectable thyroglobulin antibodies ([gt]3000 IU/mL), and thyroid peroxidase antibodies ([gt]1000 IU/mL), consistent with autoimmune hypothyroidism. She was started on L-thyroxine replacement and after her thyroid function tests normalized, her edema resolved. Four years later, while still euthyroid (TSH of 4.19UIU/L, free T4 of 1.2ng/dL), she developed another episode of left ankle and foot swelling accompanied with discoloration. Work-up showed mildly increased ESR (28 mm/hour) and slightly elevated C3 complement of 125 mg/dL (normal 51-95mg/dL) with normal rheumatologic and immunologic markers. A Doppler US revealed no vascular abnormalities, and an MRI showed effusion and soft tissue inflammation. She was diagnosed with arthritis and responded to NSAID and physical therapy. Over the next year, she developed discrete areas of atrophy on both calves that eventually coalesced. Repeat MRI revealed scattered areas of subcutaneous fat atrophy. Additional autoimmune work-up showed mildly detectable GAD 65 (1.6 U/ml) and undetectable IA-2, insulin, ovarian, 21 hydroxylase antibodies and thyroid-stimulating immunoglobulins. Biopsy identified LP with accumulation of CD3 positive T-cells and macrophages, along with marked fibrosis.[br]Conclusion: This is an unusual case of recurrent LP in association with autoimmune hypothyroidism and possible autoimmune insulinitis. We are in the process of performing additional immunological studies in the affected adipose tissue in order to determine whether LP is of autoimmune origin.[br][br]Nothing to Disclose: ZS, SL, HGM, WJD, FDL, J-PDC, EAS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1157 317 2468 MON-595 PO54-02 Monday 2137 2012


2133 ENDO12L_MON-596 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) A Case of Combined Pituitary Hormone Deficiency Due to POU1F1 Mutation Resembling Achondroplasia Akira Endoh, Hideo Sugie Hamamtsu Medical Center, Hamamatsu, Japan; Jichi Medical University, Shimono, Japan Background; Achondroplasia is characterized by frontal bossing, midface hypoplasia, otolaryngeal system dysfunction, and short stature. The common complication of hydrocephalus caused by the narrowed jugular foramina foramen magnum stenosis occurs even in the early infant. Combined pituitary hormone, including GH, TSH deficiency (CPHD) also presents growth failure from the early infant period, but not hydrocephalus.[br]Clinical Case:The affected boy was born with normal birth weight and length. Neonatal TSH-based neonatal screening was normal. At 3 months of age he was referred to our unit because of early growth failure. Body length SDS was -4.18 and frontal bossing, depressed nasal bridge, micro gnathia, short arms also were observed. He did not exhibit generalized hypotonia. Head CT showed hydrocephalus due to small foramen magnum. A clinical initial diagnosis of achondroplasia was made. He was followed for height, weight, and head circumference using growth curves standardized for achondroplasia. At the age of 11 months, he showed appetite loss and setting sun sign. Serial magnetic resonance (MRI) imaging demonstrated mild compression of the medulla oblongata by the thickened posterior rim foramen magnum. That was typically observed in achondroplasia. Foramen magnum decompression was performed. At the age of 14 months, his growth gradually decelerated even compared to achondroplasia. The first laboratory results showed the combination of a very low free T[sub]4[/sub] (2.2 pmol/L, [normal, 10.4-26]), normal TSH (1.61 mU/L, [normal, 0.35-4.94]) and low insulin-like growth factor I ([lt]4 mg/L). Thyroid hormone replacement was commenced. In spite of thyroxin replacement, he showed severe growth retardation. At the age of 3 yr, further investigation was performed. Standard provocation test revealed TSH and GH were lacked, but LH, FSH, ACTH were intact. MRI of the brain demonstrated a small anterior pituitary gland with normal posterior lobe. GH replacement therapy was started after this time, and he responded well. Sequence analysis for POU1F1 revealed a dominant negative mutation in codon 271 (R271W), but for FGFR3, the gene responsible for achondroplasia demonstrated no mutation.[br]Clinical Lesson: This is the first case in the early infant with CPHD due to POU1F1 mutation demonstrating hydrocephalus with small foramen magnum resembling achondroplasia.[br][br]Nothing to Disclose: AE, HS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1077 317 2469 MON-596 PO54-02 Monday 2138 2012


2134 ENDO12L_MON-597 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Use of Tolvaptan, a Selective Oral Vasopressin V[sub]2[/sub]-Receptor Antagonist in a Pediatric Patient with SIADH Ashwini Mallappa, Sowmya Krishnan University of Oklahoma Health Sciences Center, Oklahoma City, OK [bold][underline]Background:[/underline][/bold] Hyponatremia is the most common electrolyte abnormality encountered in clinical medicine. Syndrome of inappropriate antidiuretic hormone secretion (SIADH) (due to head trauma, brain tumor, and post hypothalamic-pituitary surgery,medications) is an uncommon cause of hyponatremia in children.[br]Current treatment options for SIADH (fluid restriction, lithium, demeclocycline, lasix) have their own limitations with suboptimal outcomes. The discovery and recent approval of Arginine Vasopressin (AVP) V[sub]2 [/sub]receptor antagonists (Vaptans) for clinical use is a new therapeutic option. Vaptans competitively bind to the V[sub]2[/sub] receptors and prevent the antidiuretic action of AVP. The efficacy and safety in pediatric population is yet to be established.[br][bold][underline]Clinical Case:[/underline][/bold] An 8 y/o girl presented for evaluation of hyponatremia. On admission, she was not tachycardic, blood pressure was elevated [gt]95% ile, and well hydrated.[br]Evaluation revealed normal Free T4/TSH and morning cortisol levels. Serum chemistries revealed hyponatremia with Na of 121mEq/L (135-145), a low serum Osmolality of 260mOsm/Kg (280-300). Urinary osmolality was inappropriately elevated at 596mOsm/Kg (300-900); diagnostic of SIADH. A lesion suspicious of low grade glioma was detected in the hypothalmic region on brain MRI.[br]She was initially treated with Lasix, fluid restriction to 1L/day and salt replacement. Sodium (Na) levels ranged from 121-124mEq/L. The family expressed constant frustration with the fluid restriction regimen and with Na levels trending further down (120-122 mEq/L), Tolvaptan was started. Therapy was initiated at a dose of 7.5mg once daily as an inpatient. Na levels increased from 127meq/L to 130meq/L in 24hrs. Tolvaptan therapy was well tolerated. The fluid restriction was increased to 2L/day. Fluid restriction continues to be a challenge. Primary polydipsia and possible resetting of her osmostat led to the excessive fluid consumption. In the last few months, the dose of Tolvaptan was increased to 15mg, then to 22.5mg with salt replacement and weekly sodium checks.[br][bold][underline]Conclusion:[/underline][/bold] Currently available treatment options for SIADH are suboptimal. The novel medication, Tolvaptan, seems to be a promising option. Tolvaptan was tolerated well in our patient and resulted in improvement in her clinic status as well as her quality of life. The need to increase the dose over the past year due to possible downward resetting of the osmotic threshold for thirst is challenging and needs close monitoring.[br][br]1.Verbalis JG, Vasopressin V2 receptor antagonists. J Mol Endocrinol. 2002 Aug; 29(1):1-9. 2.Smith D et al., Downward resetting of the osmotic threshold for thirst in patients with SIADH. Am J Physiol Endocrinol Metab. 2004 Nov; 287(5):E1019-23. 3.Schrier RW et al., Tolvaptan, a selective oral vasopressin V2-receptor antagonist, for hyponatremia. N Engl J Med. 2006 Nov 16; 355(20):2099-112.[br][br]Nothing to Disclose: AM, SK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1910 317 2470 MON-597 PO54-02 Monday 2139 2012


2135 ENDO12L_MON-598 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) A Case of Congenital Nephrogenic Diabetes Insipidus (CNDI) Yusuke Mizuno, Akiko Yamamoto, Satsuki Nishigaki, Masahiro Noda, Kengo Miyashita, Yasuhiro Naiki, Reiko Horikawa National Center for Child Health and Development, Okura, Setagaya-ku, Japan Background:[br]Nephrogenic diabetes inspidus (NDI) is a rare inherited disease characterized by an inability to concentrate urine in response to AVP. Patients have severe polyuria, nocturia and polydipsia. Patients sometimes show dehydration with hypernatremia, failure to thrive, irritability and fever. The disorder is caused by functional defects in the AVPR2 gene (AVPR2) encoding the vasopressin V2 receptor (90% of the cases) or aquaporin-2 gene (AQP2) encoding the AVP-responsive water channel in collecting tubules of the kidneys (10% of the cases). Diagnosis is made by measurement of urine volume, serum Na and ADH levels, urine and plasma osmolarity, and water restriction test with AVP administration. In some cases, however, the responsiveness to AVP can be observed even the other tests suggest the diagnosis to be NDI. Here we report a Japanese male patient with NDI, who partially responded to treatment with DDAVP.[br]Case :[br]A 1-year-4-months old boy presented to our hospital with complaint of severe polydipsia for 6 months. His prenatal history was unremarkable. He had no family history of NDI, and his parents were not consanguineous. He had polyuria and polydipsia. He became irritable when water access was limited.[br]On admission, his height was 79.8cm(+0.3SD) and body weight was 8.37kg(-1.8SD). His mental and moter development was normal.Blood pressure was 90/44mmHg and pulse rate was 98/min. Daily urine output was more than 2500ml([gt]5800ml/m[sup]2[/sup]SA)Urinary and plasma osmolalities were 310mOsm/l and 285mOsm/l, respectively, when plasma ADH level was 35pg/ml. Serum Na 140mEq/l, K 4.6mEq/l, BUN 15.6 mg/dl, and Cr 0.25 mg/dl.[br]After 7-hour of water deprivation test with 5% body-weight-loss, urine osmolality elevated up to 394mOsm/l. Plasma ADH level was 82pg/ml. After administration of 5 [micro]g DDAVP, urine osmolality elevated only to 513mOsm/l, suggesting his condition as partial nephrogenic DI. We perform high concentration sodium loading test for confirmation. Urine osmorality increased to 650mOsm/l with serum osmolality of 306mOsm/l while plasma ADH was 490 pg/ml.[br]Brain MRI showed no abnormality, and abdominal ultrasonograph revealed no evidence of obstructive nephropathy including hydronephrosis.[br]Since partial urine concentration with highly elevated plasma ADH levels, we diagnosed him as having partial nephrogenic DI, although he partly responded to DDAVP administration. Genetic investigation for AVPR2 and AQP2 is on its process.[br][br]Nothing to Disclose: YM, AY, SN, MN, KM, YN, RH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1450 317 2471 MON-598 PO54-02 Monday 2140 2012


2136 ENDO12L_MON-599 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Familial Neurohypophyseal Diabetes Insipidus Due to a Mutation in the Arginine Vasopressin-Neurophysin II Gene Chan Jong Kim, Eun Mi Yang, Young Jong Woo Chonnam National University Medical School [amp] Hospital, Gwangju, Korea Background: Autosomal dominant familial neurohypophyseal diabetes insipidus (adFNDI) is an inherited disorder of free water conservation characterized by childhood onset polyuria, polydipsia and dehydration caused by arginine vasopressin deficiency. A variety of disease-causing mutations of the arginine vasopressin neurophysin II gene (AVP) on chromosome 20p13 have been described. We report a Korean kindred in whom adFNDI is associated with a mutation in exon 2 of the AVP gene encoding the neurophysin II moiety.[br]Clinical case: A four year old body with polyuria and polydipsia was shown to have central diabetes insipidus using the water deprivation test and a vasopressin challenge test. His family history was consistent with autosomal transmission of the polyuric syndrome, with affected family members in three generations, including several females. Direct sequencing of the AVP gene showed a heterozygous missense mutation in exon 2 of the AVP gene (Cys98Gly). This mutation was predicted to yield an abnormal AVP precursor in its neurophysin II moiety and the function of neurophysin as a carrier protein for AVP would be impaired. The proband[apos]s mother and additional three family members have the same mutation. Presence of this mutation suggests that the portion of the neurophysin peptide encoded by this sequence is important for the appropriate expression of vasopressin.[br]Conclusion: We present a mutation of AVP in a Korean family suffering with adFNDI over three generations.[br][br]Nothing to Disclose: CJK, EMY, YJW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 33 317 2472 MON-599 PO54-02 Monday 2141 2012


2137 ENDO12L_MON-600 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Growth Hormone Deficiency in Primary Ciliary Dyskinesia: A Case Report Swapna Dharashivkar, Stephen J Winters, Susan Raghavan Univesity of Louisville, Louisville, KY; Children[apos]s Diabetes and Endocrinology, Louisville, KY Chronic inflammatory conditions are known to cause growth failure. Typically, the clinical focus is on the inflammatory disease while the diagnostic evaluation and treatment of the growth failure is delayed. GH secretion is usually normal and IGF-1 is low. We present a child with Primary Ciliary Dyskinesia (PCD) and GH deficiency in whom GH therapy was highly effective.[br]A Caucasian girl (46, XX) presented at age 8 yrs with height [lt]-2.25 SDS. She was diagnosed with PCD at age 7, by nasal mucosal electron microscopy following recurrent respiratory and urinary tract infections. Born at 27 weeks, birth weight was 900 gms (IUGR). By age 2, catch-up growth was excellent with height at the 25th percentile and weight at the 3rd. By age 9, she had fallen below -2.25 SDS for height, with a delayed bone age (7 yrs). She had recurrent infections in spite of prophylactic antibiotics and medical and surgical treatment. TSH 1.97 uIU/mL, Free T4 1.23 ng/dL, AM cortisol 6 [mu]g/dL (2.4-22.9), were normal. IGF-1 41 ng/mL (140-308), IGF-BP3 0.6 mg/L (1.8-7.1) and peak GH of 2.2 ng/mL following glucagon and arginine stimulation implied GH deficiency. PFT, DEXA scan and pituitary MRI were normal. rhGH therapy was begun at 9.5 yrs (0.2 mg/kg/wk). Within 6 mos growth velocity increased from 1.5 to 7.2 cm/yr, IGF-1 to 445 ng/mL, IGF-BP3 to 5.9 mg/L and bone age to 7 10/12 yrs at chronological age 10 1/12 yrs. Notably, 3 yrs of rhGH replacement was associated with a sustained 45% decrease in infection rate from 9 ([plusmn]4.3)/yr to 5/yr with no change in serial PFTs.[br]Studies in chronic inflammatory diseases have shown disturbance of GH/IGF-1 with reduced IGF-1 and IGF-BP3 levels and resistance to GH. Few cases of GH deficiency have been reported. Pro-inflammatory cytokines e.g. IL-6 decrease IGF-1 half-life and inhibit hepatic GH signaling. TNF[alpha] reduces hepatic expression of GHR. IL-1[beta] and TNF[alpha] blunt the IGF-1 mRNA response to GH, and GH resistance induced by IL-1[beta] might be mediated by decreased GHR mRNA. GH improves immune function by various mechanisms including increased antibody production and neutrophil adhesion. Short-term studies of GH treatment in Crohn[apos]s disease and juvenile idiopathic arthritis have been associated with improved growth rates.[br]rhGH replacement in our case improved growth and appears to have decreased the rate of infections. Our experience with this child suggests that children with growth failure due to recurrent infections be evaluated and treated for GH deficiency.[br][br]Nothing to Disclose: SD, SJW, SR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 667 317 2473 MON-600 PO54-02 Monday 2142 2012


2138 ENDO12L_MON-601 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) A Novel Homozygous Growth Hormone Receptor ([italic]GHR[/italic]) Mutation, [italic]c.266+83G[gt]T[/italic], Generates a New Activating Donor Splice Site in Intron 4, Resulting in a Classical GH Insensitivity Phenotype Eva Feigerlova, Michael A Derr, Mike Swinyard, Jeannie Farnsworth, Ron G Rosenfeld, Vivian Hwa Oregon Health [amp] Science University, Portland, OR; Centre Hospitalier R[eacute]gional et Universitaire, Vandoeuvre-l[egrave]s-Nancy, France; Mountain Vista Medicine, South Jordan, UT Mutations in the human GH receptor ([italic]GHR[/italic]) gene are the most common causes for growth hormone insensitivity (GHI) syndrome and IGF-1 deficiency (IGFD). The preponderance of [italic]GHR[/italic] mutations is located in the GH-binding extracellular domain (ECD), encoded by exons 2 to 7. The ECD can be proteolytically cleaved to circulate as GH binding protein (GHBP). Lack of detectable GHBP can indicate a defective [italic]GHR[/italic] gene.[br][bold]Objective:[/bold] Evaluate the cause of severe growth retardation in three siblings who exhibited classical GHI (Laron) phenotypes.[br][bold]Case Reports:[/bold] Two brothers (ages 16.5yr, 14.9yr) and their half-brother (11.3yr) presented with extreme short statures: heights were 112.4cm (-7.05 SDS), 106.1cm (-6.34), and 93.8cm (-8.02 SDS), respectively. Parents were consanguineous and of normal stature, as was a full brother of the two older boys. Basal serum GH levels were normal or elevated, while serum concentrations of IGF-I and IGFBP-3 were below detection. Significantly, serum GHBP was also below detection. A diagnosis of GHI syndrome was made and IGF-I therapy initiated, with good responses.[br][bold]Results:[/bold] The clinical presentation and undetectable serum GHBP levels, concomitant with abnormally low circulating IGF-I and IGFBP-3, suggested a defect in GHR. Molecular analysis of the [italic]GHR[/italic] gene in all 3 probands, however, revealed normal exonic and exon-intron junction sequences. Homozygous deletion of exon 3, a common polymorphism, was noted. Interestingly a novel single nucleotide change, G[gt]T, within intron 4 at position [italic]c.266+83[/italic], was also noted. Upon further analysis, the [italic]c.266+83G[gt]T[/italic] creates an active donor splice site that is preferentially utilized, and leads to retention of 81 intronic nucleotides in the final [italic]GHR[/italic] cDNA product (from established fibroblasts of the two full siblings). The consequence is a translational frameshift, terminating 19 amino acid residues within the new retained sequences. The mother is heterozygous for the exon 3 deletion and [italic]c.266+83G[gt]T[/italic], whereas the normal statured brother has full-length wild-type [italic]GHR[/italic] sequences (father was unavailable for analysis).[br][bold]Conclusion:[/bold] We report a rare, novel [italic]GHR[/italic] activating splicing mutation in siblings who presented with classical GHI syndrome. The [italic]c.266+83G[gt]T[/italic] mutation in intron 4 generates a strong donor splice site that leads to early protein termination and subsequent undetectable serum GHBP concentrations. Our data emphasize the importance of analyzing [italic]GHR[/italic] cDNA when intronic nucleotides are altered.[br][br]Sources of Research Support: Tercica Inc. (to RGR). RGR has received payments for consulting or lectures from Tercica. This potential conflict of interest has been reviewed and managed by OHSU.[br][br]Disclosures: RGR: Scientific Board Member, Eli Lilly & Company; Speaker, Eli Lilly & Company. Nothing to Disclose: EF, MAD, MS, JF, VH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 891 317 2474 MON-601 PO54-02 Monday 2143 2012


2139 ENDO12L_MON-602 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Characterization of a Novel Combined Heterozygous IGF1R and SHOX Mutation of a Patient with Short Stature Eva-Maria Sophia Radermacher, Juergen Klammt, Anja Barnikol-Oettler, Michael Ranke, Gudrun Rappold, Jan M Wit, Marina Schlicke, Heike Stobbe, Wieland Kiess, Roland Pfaeffle University Hospital Leipzig, Leipzig, Germany; University Hospital Tuebingen, Tuebingen, Germany; Ruprecht-Karls-University Heidelberg, Heidelberg, Germany; Leiden University Medical Center, Leiden, Netherlands Longitudinal growth is a trait regulated by a plethora of genes. Especially, the insulin-like growth factor 1 receptor (IGF1R) plays an essential role in growth regulation. For bone development the transcription factor short statured homeobox gene (SHOX) has been proposed to be of fundamental importance.[br]Here, we report on a patient, who carries a heterozygous exon 6 deletion in the IGF1R and a heterozygous point mutation in the SHOX gene (p.Met240Ile). The appropriate for gestational age-born patient showed growth retardation at the age of six years with a height of 102.5 cm (-3.3 SDS) and weight of 15.9 kg (-2.6 SDS). The mother bearing the IGF1R deletion had an adult height SDS of -1.1. The height of the father harboring the SHOX mutation was normal (-0.9 SDS). Furthermore, patient[apos]s IGF1 levels were in low-normal range and the boy presents no additional abnormalities.[br]The IGF1R exon 6 deletion that was identified by multiplex ligation-dependent probe amplification was defined by long-range PCR to span 5.211kb. In patient[apos]s fibroblasts we revealed that nonsense-mediated decay results in IGF1R haploinsufficiency by removing mRNA of the mutated allele. Receptor haploinsufficiency was confirmed by Western blot. Accordingly, IGF1 stimulated phosphorylation studies showed a decrease in IGF1R autophosphorylation but, unexpectedly, AKT/PKB activation was not impaired.[br]Due to the fact that both mutations in the IGF1R and SHOX genes have been described to cause growth failure, we hypothesize that the pathways of the two gene products might interact.[br]In summary, we have identified the genetic cause of IGF1 resistance in a growth retarded boy and disclosed the underlying pathomechanism. Due to the unexpected activation pattern of the AKT/PKB signaling branch and a possible additive effect of the parental IGF1R and SHOX mutations, investigations to explore the interplay between both signaling pathways in fibroblasts and osteoblast cell models are in progress.[br][br]Disclosures: GR: Research Funding, Eli Lilly & Company; Speaker, Eli Lilly & Company; Collaborator, Eli Lilly & Company. RP: Medical Advisory Board Member, Eli Lilly & Company; Speaker, Ferring Pharmaceuticals, Ipsen. Nothing to Disclose: E-MSR, JK, AB-O, MR, JMW, MS, HS, WK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1021 317 2475 MON-602 PO54-02 Monday 2144 2012


2140 ENDO12L_MON-603 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) SGA Short Stature Bearing with a Nonsense Mutation (p.Q1220X) of the IGF-I Receptor Masanobu Fujimoto, Yuki Kawashima, Naoki Hamajima, Rei Nishimura, Keiichi Hanaki, Susumu Kanzaki Faculty of Medicine Tottori University, Yonago, Japan; Nagoya City West Medical Center, Nagoya, Japan The insulin-like growth factor (IGF) plays key roles in intrauterine fetal growth as well as postnatal growth via the IGF-1 receptor (IGF-1R). Heterozygous IGF-1R mutations presenting with short stature were observed in over 9 families, including our patient (Kawashima Y, 2005). Recently, we determined another case with SGA short stature bearing a nonsense mutation (p.Q1220X) of the IGF-I receptor.[br]We identified heterozygous mutation (p.Q1220X) of the IGF-1R gene in an 8-yr-old Japanese boy with SGA short stature. The boy was born at 40 weeks of gestation, with a birth weight of 2,228 g (-3.3 SD), birth height of 46 cm (-2.1 SD) and head circumference of 28.7 cm (-3.7 SD). He has no family history of short stature, and his parent and brother were not identified the mutation (p.Q1220X). At 3 years of age, he presented with missing catch-up growth; his height was 82.1 cm (-3.2 SD); head circumference, 46.0cm(-2.3SD). His IGF-1 serum level at age 3 showed extremely high [298.4 ng/ml (+3.8 SD)]. The GH stimulation test at age 3 showed a normal GH response (peak value: 15.6 ng/ml). He had symptoms of hyperactivity disorder, and his score of intelligence test (WISC-III) at age 6 was 85 (verbal IQ: 92, performance IQ 78). GH treatment was started at 6 years of age and continued for 2 years (0.23 mg[bull]kg-1[bull]week-1). He did not respond well to GH during the 2 years of treatment, in which his height velocity changed from -3.1 SD (99.5 cm) to -2.5 SD (112.7 cm). Moreover, his IGF-1 serum level showed extremely high [558.0 ng/ml (+2.8 SD) ] under GH treatment. His present height is 113.6 cm (-2.7 SDS) at the age of 8 years, 9month.[br]We identified new nonsense mutation of IGF-1R in SGA short stature with high serum IGF-1 levels and poor response to GH treatment. This mutation site in tyrosine kinase domain results in the absence of carboxyl terminal fragment of IGF-1R after the mutation site (p. Q1220). Since this lacked carboxyl fragment contains the important site for cell proliferation, we concluded this mutation resulted in IGF-1R dysfunction and short stature.[br]Several new mutated IGF-1R may be identified in patients with SGA short stature who show higher serum IGF-1 levels or are resistance to GH treatment.[br][br]Nothing to Disclose: MF, YK, NH, RN, KH, SK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 512 317 2476 MON-603 PO54-02 Monday 2145 2012


2141 ENDO12L_MON-604 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Familial Xp22.33-Xp22.12 Deletion Delineated by Chromosomal Microarray Analysis Causes Proportionate Short Stature Chahee Kwun, Sungyoon Cho, Chang-Seok Ki, Ja-Hyun Jang, Young Bae Sohn, Sung Won Park, Se Hwa Kim, Su Jin Kim, Dong-Kyu Jin Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea; Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea; Ajou University Hospital, Suwon, Republic of Korea; National Cancer Center, Goyang, Republic of Korea Patients with Xp deletions have short stature and may have some somatic traits typical of Turner syndrome, whereas gonadal function is generally preserved. In most studies of these patients, microsatellites have been used to determine the break point of the Xp deletion. In the present study, we describe the clinical, cytogenetics, and chromosomal microarray analysis of a family with an Xp22.33-Xp22.12 deletion. Two female siblings, aged 8 years 9 months and 11 years 10 months, presented with short stature. The older sibling[apos]s height(index case) was 137.9cm (-1.81 SDS) and the younger sibling[apos]s height was 118.6cm (-2.13 SDS). The mother and both daughters had only a short stature; a skeletal survey showed normal findings except for midly shortened 4th and 5th metacarpal bone. No features of Turner syndrome were present. The deletion appeared terminal with a breakpoint within Xp22.2 located about 19.9Mb from the Xp telomere. The deletion contained 102 protein-coding genes. A probe of the end breakage point was located at the 19,910,848th base of the X chromosome. Therefore, the breakage point was concluded to be located between these two probes. In summary, we report familial case of an Xp deletion. The findings of our study may be helpful in further analyzing the phenotypes associated with Xp deletions.[br][br]Nothing to Disclose: CK, SC, C-SK, J-HJ, YBS, SWP, SHK, SJK, D-KJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 510 317 2477 MON-604 PO54-02 Monday 2146 2012


2142 ENDO12L_MON-605 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Insensitivity to Growth Hormone Therapy in an Individual with Short Stature Caused by Haploinsufficiency of the Insulin-Like Growth Factor 1 Receptor Cedric Ng, Juliana Austin, Vivian Hwa, Ron Rosenfeld University of California, Irvine, CA; Long Beach Memorial, Miller Children[apos]s Hospital, Long Beach, CA; Oregon Health [amp] Science University, Portland, OR [bold]Background[/bold]: Hypothalamic-derived GHRH upregulates the release of GH from the pituitary gland, which in turn stimulates the release of IGF-1 primarily produced by the liver. IGFBP-3 and ALS bind to IGF-1 and increase its half-life. IGF-1 binds the IGF1R, a cell surface tyrosine kinase receptor, leading to growth. Any imprecision in this intricate system will cause deficiencies in normal human growth.[br][bold]Clinical Case: [/bold]DB is an 11 year old M who presented at 3 years 2 months with failure to thrive (weight 9.7 kg, -3.2 SDS) and short stature (height 83.7 cm, -3.5 SDS). He was born at 36 weeks gestation with a weight of 1.987 kg (-3.4 SDS) and length of 40.6 cm (-5.2 SDS). His mother[apos]s height is 152.4 cm (-1.9 SDS) and his father[apos]s height is 165.1 cm (-1.9 SDS). Initial laboratory evaluation included an IGF-1 of 110 ng/mL (normal range, NR, 54-178 ng/mL), an IGFBP-3 of 2.3 ng/mL (NR, 0.8-3.0 ng/mL), and a normal GH stimulation test (peak of 38 ng/mL). Prior to starting GH therapy, his growth velocity was 5.8 cm/year (-1.9 SDS). At 3 years 10 months, Somatropin was started at 0.47 mg/kg/week and it has continued to date, with dose adjustments made according to weight and IGF-1 levels. After initiation of GH therapy, his growth velocity did not improve (4.97 cm/year). At 9 years 8 months, a missense mutation in exon 6B of the SHOX gene was identified (the clinical significance is unknown). Because of his lack of response to GH therapy, additional studies were obtained. At 11 years 9 months, a heterozygous mutation in exon 18 of the [italic]IGF1R[/italic] leading to a frameshift and early protein termination was identified. The same mutation, a 19 base pair duplication in exon 18, was previously reported by our group. Given the normal height of DB[apos]s parents, his [italic]IGF1R[/italic] mutation is likely to be [italic]de novo[/italic].[br][bold]Conclusion:[/bold] Heterozygous mutations in the [italic]IGF1R[/italic] lead to short stature. To date, there are no identified cases of homozygous mutations in the [italic]IGF1R[/italic]. Homozygous mutations are likely lethal, which is supported by mouse models. This case demonstrates that the growth velocity of individuals who are haploinsufficient for the [italic]IGF1R[/italic] does not improve with GH therapy. Therapeutic options are limited as patients with [italic]IGFIR[/italic] mutations are GH insensitive and display degrees of IGF-1 resistance.[br][br]Disclosures: RR: Scientific Board Member, Eli Lilly & Company; Speaker, Eli Lilly & Company. Nothing to Disclose: CN, JA, VH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2184 317 2478 MON-605 PO54-02 Monday 2147 2012


2143 ENDO12L_MON-606 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Growth Hormone Treatment Response in a Patient with Respiratory Chain Defect Saqer Khaled Al Kharabsheh, Narjis Batool, Anzar Haider Jordan University of Science and Technology, Irbid, Jordan; Cleveland Clinic, Cleveland, OH [bold]Background:[/bold] Children with mitochondrial respiratory chain deficiency may present with growth failure.[br]Growth hormone (GH) deficiency is not well characterized in these patients and responsiveness to GH therapy remains controversial with symptoms worsening in some patients because of an increase in GH induced ATP energy demand in an already energy depleted mitochondrial defective cell.[br][bold]Clinical case:[/bold] We report a 11 year old male with mitochondrial respiratory chain deficiency with growth failure who was found to have growth hormone deficiency.[br]He was born at term with a birth weight of 5 lb 8 oz and had uneventful neonatal period.[br]He initially presented with seizures, failure to thrive, development delay and mild hypotonia at age 12 months. Serum Lactate and Pyruvate level were elevated and measured 3.7 (range 0.9-2.0 mmol/L) and 0.13 (range 0.03-0.08 mmol/L) respectively. Follow up muscle biopsy confirmed low Respiratory Chain Complex (RCC)-1 profile of 41.8 (range 89-222 Unit mU/mg); other RCC profiles were normal. Mitochondrial DNA mutation, karyotype and microarray analysis were all normal.[br]He was started on anticonvulsant medication and [ldquo]mitochondrial cocktail[rdquo] (Carnitine, Coenzyme Q 10, Vitamins B1, B2, C, E and lipoic acid) which resulted in improved muscle tone but no improvement in growth rate.[br]An endocrinological evaluation performed at age 5 year demonstrated elevated TSH of 6.07 (ref range 0.4-5.5 uU/ml) but normal free T4 of 1.3 (ref range 0.7-1.8 ng/dl). Thyroxin supplement did not improve linear growth. Further evaluation with GH stimulation study using arginine and clonidine as secretagogues at age 9 year demonstrated a low peak growth hormone value of 3.1 ng/ml (normal response is [gt]10). MRI of the head did not identify any pituitary or hypothalamic structural abnormality. IGF-1 and IGFBP-3 level were at lower normal range. GH was started at a dose of 0.05 mg/kg/day. Annualized growth velocity improved from a pre-treatment value of 3.6 cm to a post treatment growth rate of 9.6 cm. Height percentile on growth chart increased from 1.7 to a 7 percentile within 2 year. Energy level and stamina improved and IGF-1 level normalized to an upper tertile.[br][bold]Conclusion:[/bold] Patients with mitochondrial RCC-1 defect who present with growth failure should have GH/IGF-1 axis evaluated and treated with GH, if found deficient. GH therapy improves linear growth and energy homeostasis without worsening of symptoms in patients with respiratory chain complex-1 defect.[br][br]Nothing to Disclose: SKAK, NB, AH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1834 317 2479 MON-606 PO54-02 Monday 2148 2012


2144 ENDO12L_MON-607 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Effective Treatment with Mecasermin in a Child with Idiopathic Short Stature and Thrombocytopenia Associated with a Novel Runx1 Mutation (c.532A[gt]C) Ramya Kollipara, Samantha Nohava, Max Feldt University of Missouri Kansas City, Kansas City, MO; Children[apos]s Mercy Hospital, Kansas City, MO Background: Runx1 is a transcription factor that regulates the differentiation of hematopoietic stem cells into mature blood cells. Mutations in Runx1 result in a rare autosomal dominant condition with mild platelet defects and predisposition to myeloid malignancies, as high as 35%. Growth failure and mental retardation are associated findings. Cooperatively Runx1 and Runx2 regulate chondrocyte differentiation and skeletal morphogenesis. This may suggest a mechanism for altered growth. Short term efficacy of growth promoting therapy, mecasermin (Increlex), has not been described in a child with growth failure and a Runx1 mutation.[br]Clinical Case: An 8 year old male with a history of asymptomatic congenital thrombocytopenia was referred at 4 years of age for growth failure without developmental delay. Pertinent family history includes a father with idiopathic thrombocytopenic purpura and other paternal family members with bleeding disorders, leukemia and lymphoma. The patient bruises easily but has not had significant bleeding. Height at presentation was 92.4 cm (-2.1 SD) with a target height of 182 cm (+1 SD). Biochemical screening was unremarkable except for platelets of 160x10[sup]3 [/sup]mcL (150-450). Bone age was delayed by 10 months. Peak GH response was 17 ng/dl ([gt]10) on provocative testing, leading to a diagnosis of idiopathic short stature and initiation of twice daily mecasermin. His growth velocity increased reaching a current height of 125 cm (-1.1 SD) with no adverse effects during the time of therapy. He was recently referred to Hematology for pre-operative clearance for a tonsillectomy. Genetic analysis revealed a novel Runx1 heterozygous sequence variant (c.532A[gt]C) predicted to be [quot]probably damaging[quot] and [quot]not tolerated[quot]. His surgical procedure was uneventful and he had no clinical or biochemical evidence of malignancy at that point.[br]Conclusion: Runx1 mutations may be an under-recognized etiology for growth failure in children with a history of thrombocytopenia. Efficacy of growth promoting therapy for this condition is not known, however this child demonstrated improved growth velocity over a 4 year period. In patients with Runx1 mutations the risk of myeloid malignancies with use of growth hormone or mesacermin is unclear, though there is no greater associated risk in the general population. Continued collaboration with Hematology will be required to monitor for potential future risk of malignancy in patients with Runx1 mutations.[br][br]Nothing to Disclose: RK, SN, MF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1352 317 2480 MON-607 PO54-02 Monday 2149 2012


2145 ENDO12L_MON-608 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Clinical and Genetic Variability in Silver-Russell Syndrome Nuria Sanz, Consol Sanchez, Manuel Rodriguez, MariaVictoria Marcos Hospital Sant Joan de D[eacute]u, Esplugues de Llobregat, Spain; Hospital de Terrassa, Terrassa, Spain Silver-Russell Syndrome (SRS) is a genetic disorder characterized by growth failure which usually manifests in both the prenatal and postnatal periods.[br]These patients are usually diagnosed during the first year of life for presenting with distinct clinical symptoms such as macrocephalia and assymmetry in the body distribution. However, most often, the entity is underdiagnosed due to its clinical heterogeneity.[br]We describe a 4 year-old boy referred to the Endocrinology Unit because of short stature. The father[apos]s height was 178 cm (0,8 SD and the mother[apos]s height 158.1 cm (-1,8 SD). An older sister was reported to have a normal stature.[br]He was born at term after an uncomplicated pregnancy; birthweight was 2745 gr (-1,6 SD), lenght 45 cm (-3,5 SD), the head circumference 33 cm (-1,33 SD) and the body mass index (BMI) 13.3 kg/m2 (0,3 SD).[br]At age 4 years 2 months, the height was 92.3 cm(-2,7 SD), weight 12,2 kg (-2,11 SD) and BMI 14.4 kg/m2 (0,9 SD). He presented with typical facial features (triangular face, prominent forehead, sharp pointed chin, thin lips, evident body assymmetry with right hemiatrophy and a shortening of the low right extremity by 1.6 cm.[br]Additional examinations disclosed a bone age delay of 1 yr, a normal GH response to stimulation tests, and low levels of IGF-I (36 ng/ml) with a positive response to exogenous GH in the IGF-I generation test 0.035 mg/kg/day/5 days. The molecular assessment showed a hypomethylation of the H19 gene on chromosome 11p15.5.[br]In this patient, the diagnosis of SRS was delayed by several years, despite having a history of intrauterine growth restriction and following routine postnatal paediatric controls. In SRS patients, birthweight may be within normal limits, but birth length is almost invariably affected.[br]Finally, underline that a similar phenotype may be due to genetic and epigenetic abnormalities in several chromosomes and with different pathogenesis.[br][br]Nothing to Disclose: NS, CS, MR, MM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2370 317 2481 MON-608 PO54-02 Monday 2150 2012


2146 ENDO12L_MON-609 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Normosomic Idiopathic Hypogonadotropic Hypogonadism (nIHH) Caused by a New Mutation in Fibroblast Growth Factor Receptor 1 Ghufran S Babar Children[apos]s Mercy Hospital and Clinics, Kansas City, MO Heterozygous loss-of-function mutations of the gene encoding fibroblast growth factor receptor 1 (FGFR1) have been described in patients with normosomic idiopathic hypogonadotropic hypogonadism (nIHH) (1). The male-to-female ratio is approximately 5:1. Approximately one-third of cases are familial, while the others are sporadic (2). Mutations in [italic]FGFR1[/italic] suggest that Kallmann Syndrome and nIHH are part of the same spectrum of disease (1). The lack of gonadotropins may result in micropenis, cryptorchidism and a failure of progression of sexual maturity.[br]CLINICAL CASE: The child presented at age of 13 years with the concern of delayed puberty. There was no history of acne, pubic or axillary hair development and denied lack of smell. The parents had normal puberty and intact sense of smell. His height was at the 85[sup]th[/sup] percentile and the weight was at 99[sup]th[/sup] percentile. He was also found to have gynaecomastia, un-descended testes and micropenis. His initial work-up showed a normal thyroid hormone screening, chemistry panel, lipid panel and a bone age of 14 years. His IGF-1 was 173 ng/ml (158- 792), LH: 0.6 MIU/ML, FSH: 1.6 MIU/ML and a total testosterone: 15 ng/dl. An ultrasound showed that the testes were at the lower end of the inguinal canal. He received HCG injections that caused testicular descent. He received 3 injections of testosterone 50 mg i.m to jump start his puberty. After six months his testosterone level was 8.5 ng/dl, LH: 0.4 MIU/ML, FSH: 0.7 MIU/ML. Since his puberty did not progress further work showed a normal XY karyotype and normal screening for KAL1 gene analysis for X-linked Kallmann Syndrome (KAL1) (Xp22.3). Furthermore, FGFR1 gene analysis for Autosomal Dominant Kallmann Syndrome (KAL2), showed that he is heterozygous for a variant of unknown significance, c.1927G[gt]A (p.Gly643Ser). He is currently receiving testosterone therapy and has adequate progression in puberty. Conclusions: To our knowledge this is a new novel mutation linked with FGFR1 gene causing normosomic idiopathic hypogonadotrophic hypogonadism.[br][br](1) Pitteloud N, Quinton R, Pearce S, et al. Digenic mutations account for variable phenotypes in idiopathic hypogonadotropic hypogonadism. J Clin Invest. Feb 2007; 117(2):457-63. (2) Waldstreicher J. Seminara S. B. ,Jameson J. L. , Geyer A. ,Nachtigall L. B. , Boepple P. A. , Holmes L. B. , Crowley W. F., Jr; The genetic and clinical heterogeneity of gonadotropin-releasing hormone deficiency in the human. J Clin Endocrinol Metab 81:4388[ndash]4395 (1996) J. Clin. Endocrinol. Metab 81:4388[ndash]4395.[br][br]Nothing to Disclose: GSB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2070 317 2482 MON-609 PO54-02 Monday 2151 2012


2147 ENDO12L_MON-610 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Gonadotropin-Releasing Hormone Agonist-Resistant Central Precocious Puberty in a Patient with a Hypothalamic Hamartoma: Successful Treatment Using Ketoconazole and Spironolactone Anna Ryabets-Lienhard, Erin Shih, Wendy G Mitchell, Mark Krieger, Mitchell E Geffner Children[apos]s Hospital Los Angeles, Los Angeles, CA; Children[apos]s Hospital Los Angeles, Los Angelese, CA; Children[apos]s Hospital Los Angeles, Los Angeles, CA [bold]Background:[/bold] Central precocious puberty (CPP) due to a hypothalamic hamartoma (HH) is a rare cause of gonadotropin-dependent precocious puberty, the usual first-line treatment for which is a long-acting gonadotropin-releasing hormone agonist (GnRHa). A surgical approach is usually reserved for the rare non-responsive cases. Herein, we describe successful treatment of CPP using ketoconazole and spironolactone in a young boy with an intrahypothalamic hamartoma who failed GnRHa treatment.[br][bold]Clinical Case:[/bold] A 4.6 yr-old boy presented with gelastic seizures, aggressive behavior, and precocious puberty including increased penile size, 6-8 mL testes, facial hair, acne, and tall stature relative to his mid-parental target height (+0.71 SD vs. +0.06 SD, respectively). Testing revealed: LH 0.93 mlU/mL, FSH 0.82 mlU/mL, testosterone (T) 242 ng/dL, and bone age (BA) 8.5 yr (yielding a predicted adult height of 152.5 cm, -3.21 SD). MRI and MR spectroscopy showed a benign hypothalamic mass, most representative of a HH. A histrelin implant was placed; however, after 4 mo, his LH was 1.90 mlU/mL and T 420 ng/dL, leading to the addition of leuprolide (ultimately at a dose of 15 mg/mo). Nonetheless, after 3 months of combined high dose GnRHa treatment his pubertal hormones remained non-suppressed (LH 1.27 mlU/mL and T 302 ng/dL), and his BA advanced (10 yr at actual age 5.33 yr). Due to the intrahypothalamic location of the HH, surgery was deemed too risky. A trial of ketoconazole (200 mg bid) and spironolactone (100 mg/day) was begun. The former has been used to treat GnRH-independent precocious puberty due to testotoxicosis, by blocking steroid synthesis in the adrenals and gonads. Serum T levels decreased to 18 and 16 ng/dL at 1 and 2 mo post-treatment respectively, with no change in BA and no hepatic or other side effects. However, his aggressive behavior continued on levetiracetam with poorly controlled seizures resistant to several anticonvulsants.[br][bold]Conclusion:[/bold] This unique case demonstrates that GnRHa-resistant CPP may occur in the setting of a HH. We have also shown that a treatment regimen typically employed for GnRH-independent precocious puberty (ketoconazole and spironolactone) has at least short-term efficacy and may obviate the need for surgical treatment of HH. To our knowledge, this is the first report using this regimen in a child with CPP. Its long-term efficacy and safety remain to be established.[br][br]Nothing to Disclose: AR-L, ES, WGM, MK, MEG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1691 317 2483 MON-610 PO54-02 Monday 2152 2012


2148 ENDO12L_MON-611 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Lessons from Spontanous Puberty in 9 Girls and 3 Mothers with Turner Syndrome Florence Fellmann, Danielle Martinet, Dorothea Wunder, Michael Hauschild, Sophie Stoppa, Nelly Pitteloud, Franziska Phan-Hug Unit of Paediatric Endocrinology and Diabetology, Lausanne, Switzerland; Unit of Medical Genetics, Lausanne, Switzerland; Unit of Gynecology and Obstetrics, Lausanne, Switzerland; Univerity of Lausanne, School of Biology and Medicine/FBM, Lausanne, Switzerland [bold]Background:[/bold] 30% of girls with Turner syndrome (TS) present with spontaneous initiation of puberty yet only 5% have menarche and 2% spontaneous pregnancy. Monosomie for X-linked genes, which may escape X-inactivation, could account for this ovarian failure. The aim of this study is to examine puberty longitudinally in a cohort of TS patients.[br][bold]Methods: [/bold]30 TS patients were evaluated retrospectively. Subjects were phenotyped in terms of spontaneous pubertal development and genotype was reviewed. Family members harboring the same chromosomal defect were also evaluated in terms of pubertal history.[br][bold]Results: [/bold]Of 30 TS patients ([gt]15yrs by 2010), 9 (30%) had spontaneous menarche followed by menstrual cycles. Breast development in 4 of these patients corresponded to Tanner stage 5 in one to Tanner stage 4. Three of them have mosaic karyotypes (45,X[30]/46,XX[70]; 45,X[93]/47,XXX[ 7 ]; 45,X[60]/46,XX[40]) and had menarche between 12-13.5 yrs with regular menses thereafter. One patient had complete puberty while her mother had two normal pregnancies, and both mother and daughter harbor the same deletion at the distal part of Xp (46,X,del(X)(p22.11)). Two patients developed ovarian insufficiency 1-2.5 yrs after menarche and were found to harbor deletions and recombinations of the X chromosome including the POF1 and POF3 regions. These two patients never received growth hormone (GH) treatment. One patient (45,X[69]/47,XXX[ 31]) developed amenorrhea in the setting of a prolactinoma at age 15. Surprisingly, two patients with 45,X karyotype showed complete puberty and normal serum gonadotropin levels yet decreasing anti-m[uuml]llerian hormone (AMH) levels following discontinuation of GH treatment.[br][bold]Clinical lessons and conclusion: [/bold]Herein we report in 30 % of TS girls spontaneous menarche, a rate much higher than previously reported (1). The observation of decreasing AMH levels following cessation of GH treatment in 3 patients suggests a potential role of GH on ovarian function. The deletion of known POF gene regions is associated with early ovarian failure yet does not appear to underlie absent pubertal development. The presence of spontaneous pubertal development in TS women with 45,X karyotype points to cryptic mosaicism or other eventual autosomal factors involved in puberty development. Array-CGH and whole exome sequencing may help identify genes and regions involved in pubertal onset.[br][br](1)Pasquino AM et al.,J Clin Endocrinol Metab 1997; 82:1810.[br][br]Nothing to Disclose: FF, DM, DW, MH, SS, NP, FP-H 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1723 317 2484 MON-611 PO54-02 Monday 2153 2012


2149 ENDO12L_MON-612 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) The Development of Precocious Puberty in a Child with Beckwith-Wiedemann Syndrome (BWS) Berrin Ergun-Longmire, Stephanie Savelli Akron Children[apos]s Hospital, Akron, OH [bold]Background:[/bold] BWS is the most common overgrowth disorder characterized by highly variable clinical findings. The cardinal features are macrosomia, abdominal wall defects, macroglossia, hyperinsulinism, hypoglycemia, facial nevus flammeus, and abdominal wall defects. It has an increased oncological risk, with an overall incidence of 10% in the first decade of life. Although urogenital abnormalities are common, precocious puberty has not been reported as a clinical feature of BWS.[br][bold]Clinical Case: [/bold]The patient is a 2 [frac12] year old girl with BWS. She has hypomethylation of LIT1 mutation. Her karyotype is 46,XX. She was born at 41 weeks gestational age. Her birth weight was 4300-gram and birth length was 54.6 cm. At birth, she had clinical features consistent with Beckwith-Wiedemann syndrome, including macroglossia, nevus flammeus markings on bilateral eyelids and upper nose, and the linear creases on bilateral ear lobes, diastases recti, and small umbilical hernia. Her blood sugar was 37. She had a palpable large abdominal mass due to right adrenal hemorrhage which resolved later. After her blood glucose was stabilized by IV dextrose she was started on diazoxide. At 4 months of age, she developed bilateral stage V Wilms tumor and and underwent chemotherapy followed by bilateral partial nephrectomy. She had tongue reduction at 19 months of age. At 2 years of age, she developed Tanner Stage 2 breast tissue. Her AFP was slightly elevated at 12.4 (ref. range: [lt]11). However, her CT of abdomen was stable; she had normal appearance of uterus and adnexes. There was no evidence of hepatoblastoma or any other tumor development. She had normal thyroid function, cortisol and IGF-1 levels. Her leuprolide stimulation test showed basal FSH and LH of 6.2 IU/L and 0.1 IU/L, respectively. Her stimulated FSH and LH were 20.6 IU/L and 1.2 IU/L, respectively. Her basal estradiol was 2.8 pg/mL and post leuprolide estradiol was 21 pg/mL. Although her leuprolide stimulation test was suggestive of early stages of puberty, her pubertal development continued to progress. Although MRI was ordered, the results are not yet available.[br][bold]Discussion: [/bold]Urogenital abnormalities including clitoromegaly and cryptorchidism are common in children with BWS. Although the decreased fertility noted in carrier males, precocious puberty has not been documented in either males or females with BWS.[br][bold]Conclusion:[/bold] This is the first report of an association between central precocious puberty and BWS.[br][br]Nothing to Disclose: BE-L, SS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 939 317 2485 MON-612 PO54-02 Monday 2154 2012


2150 ENDO12L_MON-613 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Yolk Sac Tumor as a Cause of Vaginal Bleeding in a 15-Month-Old Girl Melissa Ann Carlucci, Marielisa Rincon, Avery Mixon Children[apos]s Hospital at Erlanger, Chattanooga, TN; Children[apos]s Hospital at Erlanger, Chattanooga, TN [bold]Background[/bold]: Early puberty is a common reason for referral to a Pediatric Endocrine clinic. The differential diagnosis includes central (pituitary based) and peripheral (non-pituitary based) causes of pubertal development. Presence of a tumor in the sacrococcygeal region of an infant typically represents a germ cell tumor, accounting for approximately 1% of childhood cancers. Endodermal sinus (yolk sac) tumors are the most common pure malignant germ cell tumors in childhood.[br][bold]Clinical Case: [/bold]A 15 month old girl was referred to the Pediatric Endocrinology clinic with a chief complaint of 1 month of vaginal bleeding. She had no report of thelarche or adrenarche. Prior history was significant only for a strawberry hemangioma on the neck. Prior evaluation included normal gonadotropins (FSH 2.1 mIU/ml, LH [lt]0.2 mIU/ml) and normal estradiol ([lt]5 pg/ml). Physical exam revealed a normal appearing toddler with length at the 50% and weight at the 90%. She had tanner I breast development, tanner I pubic hair, no axillary hair and serosanguineous discharge from the vaginal introitus with mild labial adhesions. No caf[eacute]-au-lait spots were noted. Ultrasound of the pelvis was ordered demonstrated an 8.9 X 4.2 X 4.6 cm vaginal mass. The patient was then referred to Pediatric Hematology-Oncology where the mass was confirmed with CT scan. A transvaginal core needle biopsy showed tissue staining strongly with AFP (alpha- fetoprotein) and serum AFP was 100,300 ng/mL, confirming the diagnosis of yolk sac tumor. The patient was started on quadruple chemotherapy consisting of bleomycin, etoposide, cisplatin and cyclophosphamide. During the first round of chemotherapy, part of the tumor was expressed vaginally, a very rare occurrence. Prior to Cycle 2 chemotherapy, serum AFP has decreased to 2020 ng/mL.The patient continues to receive chemotherapy and is doing well clinically with an excellent prognosis.[br][bold]Conclusions[/bold]: This case illustrates the need to consider alternative causes of vaginal bleeding in patients referred for precocious puberty. The lack of any other physical evidence of puberty in the context of vaginal bleeding must direct the judicious clinician towards possible non endocrine etiologies.[br][br]Nothing to Disclose: MAC, MR, AM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 167 317 2486 MON-613 PO54-02 Monday 2155 2012


2151 ENDO12L_MON-614 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Sertoli Cell Testicular Nodules with Prepubertal Gynecomastia and Incomplete Peutz-Jeghers Syndrome Phenotype Julia R Broussard, Figen Ugrasbul Children[apos]s Mercy Hospital and Clinics, Kansas City, MO Sertoli cell testicular tumors in males with Peutz-Jeghers syndrome (PJS) have been recognized as a cause of prepubertal gynecomastia. Mutations in the tumor suppressor gene LKB1/STK11 were described in 50-90 % patients with PJS. In the past the treatment was orchidectomy, but pharmacologic therapy has increasingly becoming a reasonable option. A 4 year old male presented with bilateral gynecomastia of 12 months duration. Parent used lavender bubble baths until 4 months prior to the visit and after stopping it, breast size decreased. Physical exam: prepubertal male, breast size 3 cm right side and 2.5 cm left side. Workup for gynecomastia was normal, except for advanced bone age of 6 years 0 months for a chronologic age 4 years 0 months. Ten months later, patient had evidence of growth spurt, testicular enlargement (early Tanner II), increase in breasts size, freckling of his lower lip and mucosa of the lower lip. No gastrointestinal problems. Precocious puberty workup was negative. Workup for adrenal pathology as a cause of gynecomastia was negative. Family history: a few relatives on mother[apos]s side of the family have freckles of their lips, maternal great-grandmother died of stomach cancer. Scrotal ultrasound (US): abnormal, inhomogeneous echo texture of both testes with prepubertal testicular volume. There was microlithiasis bilaterally including a punctuate area of calcification in the right testicle. Sertoli cell nodules of the right testicle were diagnosed by histological exam. Presumably, both testes have Sertoli cell nodules based on the US. Inhibin A=311 pg/mL [[lt]10], inhibin B[lt]10 pg/mL [40-270]. To improve gynecomastia and delay skeletal maturation, treatment with aromatase inhibitor Anastrozole was started. Genetic testing for Peutz-Jeghers syndrome showed heterozygous partial deletion of the STK11 gene. Patient[apos]s esophago/gastro/intestinal biopsies were within normal limits. While on Anastrozole therapy, patient[apos]s bone age was 7 years 0 months for chronologic age 5 years 7 months (borderline advanced). Biochemical profile showed inhibin A=165 pg/mL and inhibin B[lt]10 pg/mL. Serial testicular US: stable findings. Anastrozole was stopped after 3.5 years of treatment, as gynecomastia resolved. Patient is being followed conservatively. Conclusion: A low incidence of invasive Sertoli cell tumors were reported in PJS and such, a conservative management with aromatase inhibitor and close follow-up is justified in noninvasive lesions, rather than surgery.[br][br]Nothing to Disclose: JRB, FU 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 587 317 2487 MON-614 PO54-02 Monday 2156 2012


2152 ENDO12L_MON-615 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) Effective Use of Bicalutamide in an Adolescent with Panhypopituitarism for Testosterone-Induced High Flow Priapism Max Feldt Children[apos]s Mercy Hospital, Kansas City, MO Background: High Flow Priapism (HFP) is an infrequent complication of exogenous testosterone supplementation. If detumescence is not quickly achieved, vascular damage and impotence are devastating complications. The complex etiology of priapism involves disruption of neuronal, vascular or endocrine pathways. Although the mechanism is not fully understood, androgens have been implicated as an important etiologic factor. Anti-androgens carry their actions by direct suppression of the penile androgen receptors. There is no consensus on the optimal course of therapy, but prior studies show the use of bicalutamide, an androgen receptor antagonist, as an effective adjunct therapy for HFP in adult subjects. Its use in pediatrics for HFP has not been previously reported.[br]Clinical case: A 14 year old male with panhypopituitarism (hypothyroidism, adrenal insufficiency, hypogonadotropic hypogonadism, growth hormone deficiency) was admitted for prolonged priapism ([gt] 24 hours) temporally related to initiation of testosterone therapy. His initial dose of testosterone cypionate (100 mg IM) was given five days prior to admission for induction of puberty. One day prior to admission he developed a painful erection that impeded voiding. Initial therapies of IV hydration, morphine, diazepam, and pseudoephedrine were ineffective. Corporal irrigation with injection of epinephrine was also of little effect. Al-Ghorab and Quackle shunts were placed, with initial improvement but subsequent return of tumescence. Intraoperative blood gas pH of 7.29 was consistent with high-flow arterial priapism. Bicalutamide 50 mg PO daily was started on day three of admission. Baseline testosterone was 380 ng/dl (10-572). The Quackle shunt was revised on the fourth day of admission. On the seventh day of admission his penile rigidity had decreased and he was able to void without a Foley catheter. The bicalutamide was continued for a total of 10 days with no adverse effects. No further erections occurred by the two month outpatient follow up visit. Reinstitution of testosterone supplementation had not been attempted at the time of this abstract.[br]Conclusion: Blockage of the androgen receptor with bicalutamide appeared to be a safe and effective adjunct therapy for this patient. Additional studies will provide more evidence regarding bicalutamide[apos]s role in treatment of testosterone induced pediatric HFP.[br][br]Nothing to Disclose: MF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1025 317 2488 MON-615 PO54-02 Monday 2157 2012


2153 ENDO12L_MON-616 POSTER SESSION: Pediatric Endocrine Case Reports: Growth [amp] Puberty (1:30 PM-3:30 PM) A Rare Case of Pubertal Gynecomastia Due to Male Androgen Insensitivity Syndrome: A Novel Mutation and Clinical and Hormonal Findings Priya Vaidyanathan Children[apos]s National Medical Center, Washington, DC Introduction: Gynceomastia is common in pubertal boys. It is considered to be physiological affecting 2/3rd of pubertal boys and resolves spontaneously most of the time. Pathological causes of gynecomastia in this age group are rare. We report a case of a healthy 17 y/o male whose clinical exam and hormonal evaluation led to the suspicion of MAIS (male androgen insensitivity syndrome) as the cause of his gynecomastia and the resultant novel molecular finding.[br]Clinical Case: A 17 year old African American male was evaluated for gynecomastia of 3 yrs duration. Height was 178.5 cm, weight is 63 Kg. He was not obese and did not take any medications. He had a normal physical and systemic exam. Distinct features were a lack of facial hair, sparse axillary hair, mid pubertal penile length, tanner 3 pubic hair, testicular volume of 15 cc with bilateral breast development tanner 3. Pertinent labs were a LH of 14 mu/ml (n; [lt]3.7) FSH 7.7 mu/ml (n [lt]5.6), Testosterone total of 1660 ng/dl (n[lt]1000), Estradiol of 64 pg/ml(n;0-53), Karyotype 46, XY. Partial Androgen Insensitivity Syndrome (PAIS) was suspected and molecular analysis of the Androgen Receptor (AR) gene was done at GeneDX. A novel A721C missense mutation was detected in the AR gene.[br]While this mutation has not been previously identified as a cause of PAIS or MAIS, Shi X et al (2002) have studied 44 mutant androgen receptors from human prostate cancer and reported that this particular mutation is associated with reduced androgen activity. The patient has no family history of gynecomastia or infertility and therefore one could speculate that this is a new mutation. Patient was referred to the plastic surgeon and successfully underwent bilateral mastectomy.[br]CONCLUSION: This case highlights a rare cause of pubertal gynecomastia, the importance of good physical exam and the utility of checking FSH, LH, Testosterone and Estradiol in boys with atypical gynecomastia. The very high testosterone level, lack of adequate body and facial hair and a smaller testicular volume for the testosterone level were clues to the diagnosis. To our knowledge, the mutation described in this young man is a novel one and fits the phenotype of MAIS.[br][br]1. Functional analysis of 44 mutant androgen receptors from human prostate cancer. Shi XB, Ma AH, Xia L, Kung HJ, de Vere White RW; Cancer Res. 2002 Mar 1;62(5):1496-502.[br][br]Nothing to Disclose: PV 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1116 317 2489 MON-616 PO54-02 Monday 2158 2012


2154 ENDO12L_MON-617 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Longitudinal Changes in Obesity and Body Mass Index among Survivors of Childhood Malignancy Dalit Modan-Moses, Kineret Mazor-Aronovitch, Orit Pinhas-Hamiel, Gal Goldstein, Hana Golan, Dalia Waldman, Amos Toren The Edmond and Lily Safra Children[apos]s Hospital, Ramat-Gan, Israel; The Edmond and Lily Safra Children[apos]s Hospital, Ramat-Gan, Israel [bold]Background:[/bold] Increased incidence of obesity, the metabolic syndrome, and cardiovascular morbidity and mortality is observed in survivors of childhood cancer.[br]Aim: to assess the incidence of overweight and obesity in a cohort of survivors of childhood cancer, to identify potential risk factors, and to evaluate longitudinal changes in obesity during follow-up.[br][bold]Methods:[/bold] 607 patients (53.9% males) were prospectively studied in our endocrine late effects clinic. At the time of first measurement, patients were 11.3[plusmn]8.8 years old and the interval from diagnosis was 2.6[plusmn]3.3 years. 313 patients were followed for [gt]12months (mean 4[plusmn]2.5 years).[br][bold]Results:[/bold] At first measurement, 28.9% of the patients were overweight (BMI[gt]85th percentile for age), while 12.1% were obese (BMI[gt]95th percentile). These rates are more than twice the reported rates in the pediatric population in Israel. The prevalence of overweight was highest in patients with leukemia (30.8%) and brain tumors (35.3%). Prepubertal patients had a significantly (p=0.045) increased risk for being overweight. During the follow-up period, BMI-SDS of patients with brain tumors increased significantly (p=0.019), while BMI-SDS of patients with solid tumors decreased significantly (0.024). For other diagnosis groups, there was no significant change in BMI-SDS. Cranial irradiation was associated with a significantly (p=0.03) increased incidence of overweight. However, within the relevant diagnosis groups (leukemia and brain tumors) cranial irradiation was not associated with an increased incidence of overweight or obesity, neither at the beginning nor at the end of follow-up. Gender and GH deficiency were not associated with an increased risk for overweight or obesity.[br][bold]Conclusions:[/bold] Obesity is a common sequel of childhood malignancy, and is associated mainly with leukemia and brain tumors. Overweight patients remained overweight as time went by. These findings enable targeting of at-risk patients in order to prevent long-term complications including the metabolic syndrome and cardiovascular diseases.[br][br]Nothing to Disclose: DM-M, KM-A, OP-H, GG, HG, DW, AT 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1282 318 2490 MON-617 PO30-02 Monday 2159 2012


2155 ENDO12L_MON-618 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Anti-M[uuml]llerian Hormone as an Excellent Marker of Gonadal Function in Childhood Cancer Survivors Yoko Miyoshi, Noriyuki Namba, Emiko Miyashita, Yoshiko Hashii, Hideaki Ohta, Keiichi Ozono Osaka University Graduate School of Medicine, Suita, Japan; Higashitoyonaka Watanabe Hospital, Toyonaka, Japan Background: Gonadal dysfunction is one of the major problems of endocrinological late effects among childhood cancer survivors (CCSs), whereas it is difficult to predict the reproductive capacity in childhood. Several studies have recently demonstrated that anti-M[uuml]llerian hormone (AMH) is an excellent marker of gonadal function.[br]Patients and Methods: We evaluated the effects of cancer treatments on gonadal functions of 80 Japanese CCSs (58 females and 22 males) in a single-center cohort in terms of their pubertal development and serum levels of gonadotropins and AMH. Clinical diagnosis was hematological disease (n=30), brain tumor (20) and solid tumor (30). Twenty-three patients (F 14, M 9) reached adulthood. The concentrations of AMH were measured by enzyme immunometric assays (Immunotech reagents, Beckman Coulter Company) and compared to the normal values reported in JCEM 2010 (1)(2).[br]Results: Among 58 female survivors, 31 (53%) had a decreased AMH level and 16 (28%) had an increased FSH level. In females with low AMH level, FSH level was high in 17, normal in 12 and low in 2. AMH level was low in females with no breast development (8/10: 80%), no spontaneous menstruation (12/17: 71%), regular menstruation (4/23: 17%), irregular menstruation (3/3: 100%) and under estrogen replacement therapy (12/15: 80%). Among 26 females after hematopoietic stem cell transplantation (HSCT), 24 (92%) had a low AMH level and 14 (54%) had an increased FSH level. A very low AMH level ([lt]1 pmol/L) was observed in 12 females, 10 of which were after HSCT. Among 22 male survivors, AMH level was low in 6 (27%), of which 4 had an increased FSH level and 5 showed low testicular volumes. Eleven (50%) males had an increased FSH level and 13 (59%) showed low testicular volume. Among 13 males after HSCT, a low AMH level was observed in 4 (31%), an increased FSH level in 9 (69%) and low testicular volume in 11 (85%).[br]Conclusion: Measurement of AMH concentration is an useful test to detect primary gonadal deficiency in CCSs. In females, AMH was an excellent marker of ovarian reserve. AMH is an indicator even for patients with no increase in gonadotropin levels: before the onset of puberty, under the sex hormone replacement therapy and combination of gonadotropin deficiency and gonadal damage. In males, comprehensive judgment of AMH levels with FSH levels and testis volume are necessary. Low AMH in male CCSs suggested severely impaired testicular function.[br][br](1) Hagen CP et al., J Clin Endocrinol Metab 2010; 95: 5003. (2) Aksglaede L et al., J Clin Endocrinol Metab 2010; 95: 5357.[br][br]Nothing to Disclose: YM, NN, EM, YH, HO, KO 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 249 318 2491 MON-618 PO30-02 Monday 2160 2012


2156 ENDO12L_MON-619 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Prevalence of Vitamin D Deficiency in Survivors of Childhood Cancer Abha Choudhary, Joanne Chou, Glen Heller, Charles Sklar Weill Cornell/New York Presbyterian, New York, NY; Memorial Sloan Kettering Cancer Center, New York, NY [bold]Introduction: [/bold][br]Cancer survivors are at a higher risk for subsequent malignancies, cardiovascular disease and low bone mineral density, which may be affected by the 25- hydroxyvitamin D (25-OH D) status. Data on 25-OH D status among survivors of childhood cancer are sparse and confined to small series, often limited to one diagnostic group.[br][bold]Aims [/bold][br]Determine the prevalence of and risk factors for 25-OH D insufficiency in a large, diverse population of cancer survivors being followed in a survivor clinic at Memorial Sloan-Kettering Cancer Center.[br][bold]Methods[/bold][br]Retrospective chart review of survivors seen for routine long-term follow-up (January 2008 to January 2011), who underwent screening blood studies including 25-OH D levels. Subjects were excluded if treatment data were unavailable, for those with a non-cancer diagnosis who had not undergone a stem cell transplant or if they were on Rocaltrol. Electronic medical records, paper charts and cancer database were used to obtain demographics and treatment related information for each subject. 25-OH D insufficiency was defined as a level [lt] 20 ng/ml.[br][bold]Results[/bold][br]484 subjects (234 males) were evaluable. Mean age at 25-OH D testing was 12.3 years (SD= 4.9), mean age at cancer diagnosis was 5.2 years (SD= 4.35), and mean interval from cancer diagnosis to 25-OH D testing was 7.02 years (SD= 4.26). Diagnoses included brain tumors (23.6%), neuroblastoma (21%), leukemia (17.6%), soft tissue sarcomas (9.1%) and bone tumors (6.0%). The mean 25-OH D level was 25.2 ng/ml; 29% of subjects were 25-OH D insufficient. In univariate analysis; race, pubertal status and age at diagnosis were associated with 25-OH D insufficiency (p [lt]0.05 for all factors). Borderline associations were seen between 25-OH D insufficiency and seasonality (p=0.20) and exposure to total body irradiation (p=0.061). In the multivariate analysis, a model including race, pubertal status and seasonality provided a good fit for the data.[br][bold]Conclusions[/bold][br]The prevalence of 25-OH D insufficiency in these cancer survivors is high and similar to what has been described in the general population. Moreover, we were unable to identify any cancer specific variables that were associated with 25-OH D insufficiency. Interventions to improve 25-OH D status in this vulnerable population are needed.[br][br]Nothing to Disclose: AC, JC, GH, CS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 123 318 2492 MON-619 PO30-02 Monday 2161 2012


2157 ENDO12L_MON-620 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Growth after Hematopoietic Stem Cell Transplantation in Children with Acute Myeloid Leukemia Seung Joon Chung, Min Jae Kang, Young Ah Lee, Choong Ho Shin, Sei Won Yang Seoul National University Hospital, Seoul, Korea Objective: Previous studies have shown that hematopoietic stem cell transplantation (HSCT) may result in growth impairment. Disease type, differences in treatment before HSCT and treatment duration before HSCT can affect growth after HSCT and act as confounding variables. By contrast, acute myeloid leukemia (AML) patients receive HSCT during their first remission and are not treated by steroids during their relatively short period of induction therapy time.The purpose of this study was to evaluate 5 years growth after HSCT and to find factors influencing final adult height (FAH) in childhood AML patients.[br]Methods: Among 97 AML patients whom received HSCT in Seoul National University Hospital, we report 24 patients whose puberty began at least 3 years after HSCT and 19 patients who reached FAH without relapse. Medical records were retrospectively reviewed. Summary measure analysis was used to evaluate for 5 year growth after HSCT and to find statistical differences between factors. Univariate and multivariate regression analysis was performed to find factors influencing FAH.[br]Results: 1) 5-years growth after HSCT: Patients received HSCT at 4.2 years of age. Six patients received radiotherapy (RT) and chronic GVHD (cGVHD) were in 4 patietns. History of RT and cGVHD significantly impaired the first 5 years growth after HSCT. But cGVHD seems to influence to only the first 2 years growth after HSCT. Age at HSCT, gender and history of steroid use were not significantly affected 5 years growth after HSCT.[br]2) Final adult height after HSCT: Patients received HSCT at 10.1 years of age. Four patients received RT. In patients reached FAH without relapse after HSCT, only history of RT significantly reduced FAH. Age at HSCT, gender and history of steroid use were not significantly affected FAH.[br]Conclusions: Growth impairment after HSCT in AML patients might be occurred. But without RT history, growth impairment seems to be temporary and improve by catch-up growth. HSCT with conditioning regimen consists of only chemotherapy thought to be not to significantly decrease FAH. So the growth hormone treatment is seems to be not needed in non-RT patients. But in patients who received RT, catch-up growth will not be shown and eventually attain reduced FAH.[br][br]Nothing to Disclose: SJC, MJK, YAL, CHS, SWY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 904 318 2493 MON-620 PO30-02 Monday 2162 2012


2158 ENDO12L_MON-621 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Oophorectomy Versus Salpingo-Oophorectomy in Turner Syndrome Patients with Y-Chromosome Material: Current Practice Patterns Assessment Roopa Kanakatti Shankar, Thomas H Inge, Philippe F Backeljauw Cincinnati Children[apos]s Hospital, University of Cincinnati College of Medicine, Cincinnati, OH; Cincinnati Children[apos]s Hospital, University of Cincinnati College of Medicine, Cincinnati, OH [underline]Background:[/underline] Gonadectomy is recommended for Turner syndrome (TS) patients with Y-chromosome material (Y-chrom) because of an increased risk for development of gonadoblastoma and dysgerminoma. It is not known if salpingectomy should also be done concurrent with gonadectomy.[br][underline]Methods:[/underline] 1) TS patients with Y-chrom were identified from a TS database at Cincinnati Children[apos]s Hospital and their records were reviewed for genetic studies, operative reports and histopathology related to gonadectomy. 2) An electronic questionnaire was sent to all members of the International Pediatric Endosurgery Group (IPEG) to assess the practice patterns of surgeons for gonadectomy in TS patients with Y-chrom.[br][underline]Results:[/underline] Of 158 TS patients, 12 (7.6%) carried Y-chrom: 8 (45X/46XY), 3 (45X/46X, isodicentric (Y)) and 1 (45X/47XYY). Mean age at diagnosis was 6.6 yrs (range: prenatal -15) and at gonadectomy was 7.3 yrs (range 1.5-15). Gonadal dysgenesis was found in 2 patients with TS phenotype, ambiguous genitalia and inguinal gonads, who underwent feminizing genitoplasty in addition to gonadectomy. Of the other 10 patients, 3 had laparoscopic salpingo-oophorectomy, 2 had laparoscopic oophorectomy, 2 had oophorectomy by inguinal approach, 1 had two laparotomies (pelvic mass) and the remaining 2 underwent surgery at an outside hospital (unknown procedure). Histopathology revealed 3 patients with gonadoblastoma and one with a gonadoblastoma-in-situ. A teratoma and a dysgerminoma were also identified in 2 of the patients with gonadoblastoma.[br]Of 54 survey respondents (10% response rate), 57% had operated on 1 to 5 cases, 11% had done 5 to 10 cases, while the rest had not done the procedure. Laparoscopic gonadectomy was the preferred procedure (96%). While 46% opted for oophorectomy alone, 26% preferred salpingo-oophorectomy and 27% used either approach. Of surgeons who had performed [gt]5 cases, the majority favored salpingo-oophorectomy.[br][underline]Conclusions:[/underline] Y-chrom in TS patients warrants gonadectomy due to a high risk of tumors (40% in our series). Laparoscopy is the preferred approach. However, our study shows there is no consensus among an international group of pediatric surgeons regarding the removal of the Fallopian tubes. Salpingo-oophorectomy may be technically easier. There may also be a lower risk for ectopic pregnancy with assisted reproductive techniques for conception in TS patients. Further studies are needed on the risk vs. benefits of salpingo-oophorectomy in this population.[br][br]Nothing to Disclose: RKS, THI, PFB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 526 318 2494 MON-621 PO30-02 Monday 2163 2012


2159 ENDO12L_MON-622 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Analysis of Heart Magnetic Resonance Imaging Features in Patients with Turner Syndrome and Evaluation of Risk Factors Associated with Aortic Dilation and Aortic Distensibility Ah Reum Kwon, Ho-Seong Kim, Hyun Wook Chae Yonsei University College of Medicine, Seoul, Republic of Korea [bold]Purpose[/bold]: It has been reported the aortic dissection and dilation occur more frequently in Turner syndrome, and they are related to reduced life expectancy of the patients. The aim of this study is to measure and elucidate risk factors associated with aortic dilatation and distensibility in patients with Turner syndrome by analysis of imaging data based on MRI of heart.[br][bold]Methods[/bold]: Heart MRI was performed in 50 patients with Turner syndrome older than ten years of age (average age 20.3[plusmn]6.6 years). Aortic diameters were measured by heart MRI at 9 positions. Aortic distensibility was quantified by relative temporal changes in the vessel cross-sectional area throughout the cardiac cycle. Clinical manifestation, anthropometric measurements, echocardiography results, blood pressure, and metabolic parameters were analyzed.[br][bold]Results[/bold]: MRI showed abnormal findings in 42% (N=21) of patients with Turner syndrome, while echocardiography showed abnormal findings in 6% (N=3). BAV was found in 3 cases by MRI while 1 case by echocardiography. Elongation of the transverse aortic arch, which was the finding associated with progression of aortic dissection, was the most common abnormal finding in MRI (N=15, 30%), while no ETA case detected by echocardiography. Aortic dilatation was identified in two ways. First, ascending root dilation (defined as a proximal ascending aorta/proximal descending aorta [gt]1.5) was found in 34% (N=17). Second, dilated ascending aorta (proximal ascending aorta/body surface area [gt] 20mm) was found in 8% (N=4) of patients with Turner syndrome. The diameter of ascending aorta was showed positive correlation with age, weight, body surface area, and body mass index. The distensibility of ascending aorta was positively correlated with age, weight, body surface area, and systolic blood pressure.[br][bold]Conclusion[/bold]: Risk factors associated with aortic dissection or rupture are BAV, coarctation of aorta, hypertension, aortic dilation and reduced aortic distensibility. The prevalence of these factors is high in this study. In addition, age, hypertension and increased BMI can be augmented risk of aortic dissection. It is suggested that heart MRI is recommended regularly in patient with Turner syndrome to detect early changes of aortic abnormalities.[br][br]Nothing to Disclose: ARK, H-SK, HWC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1378 318 2495 MON-622 PO30-02 Monday 2164 2012


2160 ENDO12L_MON-623 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Turner Syndrome and High Blood Pressure by Ambulatory BP Monitoring Shin Mi Kim, Min Jae Kang, Young Ah Lee, Choong Ho Shin, Sei Won Yang Seoul Red Cross Hospital, Seoul, Republic of Korea; Seoul National University College of Medicine, Seoul, Republic of Korea [bold]Introduction[/bold]: Adults with Turner syndrome (TS) are at an increased risk of morbidity and mortality from cardiovascular disease. This study aimed to evaluate the prevalence of hypertension and its risk factors, and the relationship between blood pressure and aortic diameters in women with TS.[br][bold]Subjects and Methods[/bold]: Forty two patients with TS (23.7[plusmn]5.6 years; 38.1%, obese; 14.3%, type 2 diabetes mellitus; 16.7%, cardiac anomaly; and 31.0%, renal anomaly) underwent 24-hour ambulatory BP monitoring (ABPM) and echocardiography. The definitions of hypertension followed the recommendations of the American Heart Association guidelines.[br][bold]Results[/bold]: The 24-hour mean systolic or diastolic hypertension was found in 24.3% (9.5%, systolic; 21.4%, diastolic) of TS population. 66.7% of the patients had less than 10% fall in the night-time BP (non-dippers). High BP was not associated with the presence of cardiac or renal anomaly, family history of cardiovascular disease (CVD). TS patients with DM were more likely to have hypertension than those without. The HOMA-IR positively correlated with systolic BP ([italic]P[/italic] [lt] 0.003, 24-hour mean, daytime and night time) and diastolic BP ([italic]P[/italic] [lt] 0.02, 24-hour mean and daytime), independently with age, BMI and cardiac or renal anomaly. The prevalence of subjects with z scores [ge]2, representing dilation, were 6.9% for the ANN, 14.3% for the AR, 15.4% for the STJ, and 13% for the AAO. Multivariable analyses revealed several notable findings: BAV predicted larger ANN and STJ; high BP predicted a larger STJ ;older age predicted a larger AAO.[br][bold]Conclusions[/bold]: 66.7% of TS patients have an abnormal BP circadian rhythm and 35.7% of TS patients had systolic or diastolic hypertension. This study suggests that IR and hyperglycemia are associated with the presence of hypertension, independently of adiposity in patients with TS. Hypertension is a risk factor for aortic dilatation. Insulin resistance and blood pressure should be carefully monitored to prevent cardiovascular morbidity in adulthood.[br][br]Nothing to Disclose: SMK, MJK, YAL, CHS, SWY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1174 318 2496 MON-623 PO30-02 Monday 2165 2012


2161 ENDO12L_MON-624 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Metabolic Effects of Oral vs. Transdermal 17[beta] Estradiol (E[sub]2[/sub]): A Randomized Clinical Trial in Girls with Turner Syndrome Lournaris Torres-Santiago, Veronica Mericq, Martha V Taboada, Nancy Unanue, Karen Oerter-Klein, Ravinder J Singh, Jobayer Hossain, Richard J Santen, Judith L Ross, Nelly Mauras Nemours Children[apos]s Clinic, Jacksonville, FL; University of Florida, Jacksonville, FL; University of Chile and Institute of Maternal and Child Research, Santiago, Chile; Nemours Children[apos]s Clinic, Orlando, FL; University of California, San Diego, CA; Mayo Clinic, Rochester, MN; University of Virginia Medical Center, Charlottesville, VA; Jefferson Medical Center, Philadelphia, PA; Dupont Hospital for Children, Wilmington, DE [bold]Background: [/bold]Short-term transdermal (TD) administration of 17[beta] estradiol (E[sub]2[/sub])[sub] [/sub]- the principal product of the intact ovary - results in E[sub]2[/sub], estrone (E[sub]1[/sub]), and bioestrogen concentrations closer to normal and in greater suppression of LH/FSH in lower doses in girls with Turner Syndrome (TS) compared with normal (1). However, whether long-term metabolic effects of estrogen differ by route using only 17[beta]E[sub]2[/sub], while titrating E[sub]2 [/sub]to the normal range has not been well characterized.[br][bold]Objective: [/bold]To assess long term metabolic effects of the same oral vs. TD 17[beta]E[sub]2 [/sub]replacement and determine feasibility of estrogen concentration-based dosing in hypogonadal girls with TS.[br][bold]Methods: [/bold]40 girls with TS, mean age: 16.7 [plusmn]1.7 yrs, BMI: 25.1 [plusmn] 5.1kg/m[sup]2 [/sup](off GH [gt] 6mo, off any estrogen [gt] 6w prior) were randomized (N=20 each) to 17[beta]E[sub]2 [/sub]either 0.5 mg orally or 0.0375 mg TD. Doses were titrated using mean follicular/luteal phase E[sub]2[/sub] concentrations of 20 normally menstruating girls as therapeutic target (96 [plusmn] 17pg/ml) up to 2 mg orally and 0.1 mg TD. E[sub]2[/sub] [amp] E[sub]1[/sub] (LCMSMS), a recombinant cell bioassay, IGF-1, lipids, LH/FSH were measured and DXA scan and indirect calorimetry performed over 12mo.[br][bold]Results: [/bold]Data are available for 1[sup]st[/sup] 6mo in all subjects. After 17[beta]E[sub]2 [/sub]replacement, E[sub]2 [/sub]concentrations were similar between groups and comparable to controls (oral: 83 [plusmn] 14pg/ml; TD: 68 [plusmn] 14, p=0.73), E[sub]1 [/sub]was much higher in the oral group (356 [plusmn] 59pg/ml vs. 43 [plusmn] 6, p[lt]0.0001). At 6 mo changes in the oral and TD groups respectively were similar for: weight: +0.9 [plusmn] 0.6kg; +0.6 [plusmn] 0.7, BMI: +0.20 [plusmn] 0.3 kg/m[sup]2[/sup]; +0.3 [plusmn] 0.3, %fat mass: -0.6 [plusmn] 0.5%; -1.5 [plusmn] 0.5, and abdominal fat:-1.3% [plusmn] 0.7; -1.9 [plusmn] 0.8 (p=NS all). IGF-1 concentrations decreased on oral 17[beta]E[sub]2[/sub] (Median: -18ng/ml (-207, 91)) but increased after TD (+53 (-82, 103, p=0.009 between groups) with significant but comparable decreases in LH [amp] FSH concentrations and no significant changes in plasma lipids within and between groups. No differences in substrate oxidation and resting energy expenditure rates were observed within or between groups after 6mo.[br][bold]Conclusions: [/bold]Oral vs. TD 17[beta]E[sub]2[/sub] for 6mo comparably affect LH/FSH, plasma lipids and substrate oxidation rates, but IGF-I is lower and E[sub]1[/sub] higher in the oral group. However, when E[sub]2 [/sub]concentrations are titrated to the normal range the route of delivery does not affect differentially body composition in hypogonadal girls with TS. Treatment data at 12mo will soon allow us to determine if these effects are sustained.[br][br](1)Taboada M, et al. JCEM 2011;96:3502-3510.[br][br]Sources of Research Support: Genentech Center for Clinical Research in Endocrinology; Nemours Research Programs; NIH/NCRR CTSA Grant Number UL1 RR024150.[br][br]Disclosures: RJS: Consultant, Mayo Clinic Patent on Vitamin D Testing. RJS: Advisory Group Member, Pfizer, Inc.; Consultant, Teva; Advisory Group Member, Novo Nordisk. NM: Principal Investigator, Pfizer, Inc., Novartis Pharmaceuticals. Nothing to Disclose: LT-S, VM, MVT, NU, KO-K, JH, JLR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 404 318 2497 MON-624 PO30-02 Monday 2166 2012


2162 ENDO12L_MON-625 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Growth Response to Growth Hormone Therapy According to Ages in Girls with Turner Syndrome Jung Hee Ko, Hae Sang Lee, Hong Kyu Park, You Jin Kim, Jin Soon Hwang Ajou University Hospital, Suwon, Korea [bold]Objectives[/bold][br]Short stature is the most common finding in Turner syndrome patients. Improving adult final height is a challenge to patients and physicians. We investigated the clinical response to growth hormone in every year of growth hormone treatment.[br][bold]Methods[/bold][br]Retrospective medical records review of 27 patients with Turner syndrome treated with recombinant human growth hormone for more than 3 years were analyzed. Differences of changes in standard deviation score (SDS) of height were measured according to the karyotype and age at the start of treatment.[br][bold]Results[/bold][br]The response of recombinant growth hormone was observed to increase the heights of the subjects to a mean value of 0.58 standard deviation (95% confidence interval 0.28 [ndash] 0.88, p[lt]0.001) above the mean for unaffected girls at the end of 3-year administration. The increment in both of height and its standard deviation score was greatest in subjects who started the treatment before 8 years of age. The height increment was highest in the first year when the treatment was started before 12 years of age. Only in the subjects whose age were below 8 years old at the start of treatment was SDS of height increased for 3 years. The age at commencement was inversely associated with standard deviation score of height at the third year. Height for Turner syndrome growth chart demonstrated statistically significant accretion over the entire period (p[lt]0.001). Karyotype did not account for the difference in height gain.[br][bold]Conclusions[/bold][br]Early start of growth hormone administration in subjects with Turner syndrome improves height response to the treatment.[br][br]Nothing to Disclose: JHK, HSL, HKP, YJK, JSH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 504 318 2498 MON-625 PO30-02 Monday 2167 2012


2163 ENDO12L_MON-626 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Radiological Features in Patients with [italic]Short Stature Homeobox-Containing (SHOX)[/italic] Gene Deficiency and Turner Syndrome before and after 2 Years of GH Treatment Gabriel Kalifa, Christopher J Child, Christine Jones, Judith L Ross, Gudrun A Rappold, Charmian A Quigley, Alan G Zimmermann, Werner F Blum Universit[eacute] Paris V Ren[eacute] Descartes, Paris, France; Eli Lilly and Company, Windlesham, UK; Thomas Jefferson University, Philadelphia, PA; Heidelberg University, Heidelberg, Germany [bold]Background and aims:[/bold] Patients (pts) with SHOX deficiency (SHOX-D) have variable degrees of skeletal anomaly, from none to pronounced mesomelic dysplasia. In pts with SHOX-D and Turner syndrome (TS), this analysis of radiological features aimed to examine baseline differences and the effect of 2-yr GH treatment.[br][bold]Patients and methods:[/bold] In a 2-yr controlled clinical trial, 51 pts with SHOX-D were randomized to GH (SHOX-GH; N=27; 0.35 mg/kg/wk) or non-treatment (SHOX-noGH; N=24). 26 GH-treated pts with TS were included for comparison. Entry criteria: genetically confirmed SHOX-D or TS; age [ge]3yr; bone age (BA) [lt]8yr (F), [lt]10yr (M), [lt]9yr (TS); prepubertal; height (ht) [lt]3[sup]rd[/sup] %ile (or [lt]10[sup]th[/sup] %ile [amp] ht velocity [lt]25[sup]th[/sup] %ile). X-rays (hand/wrist, forearm, lower leg) were assessed at baseline and 2-yr for presence or severity of a variety of radiological anomalies.[br][bold]Results:[br][/bold][bold]Baseline:[/bold] SHOX-noGH, SHOX-GH and TS had similar mean[plusmn]SD age (7.5[plusmn]2.7, 7.3[plusmn]2.1, 7.5[plusmn]1.9 yr respectively; p=0.91), BA (6.6[plusmn]2.8, 6.5[plusmn]2.0, 6.7[plusmn]1.6 yr; p=0.93) and height SDS (-3.3[plusmn]1.0, -3.3[plusmn]0.8, -3.7[plusmn]0.9; p=0.11). Tibial tuberosities and Kosowicz sign (hypertrophic internal femoral condyle) were less common in SHOX-GH than TS (13% vs 44%; 13% vs 56%; p[le]0.03 for both). Prevalence of radial bowing for SHOX-noGH, SHOX-GH, and TS was 71%, 52% and 36%, respectively. Moderate/severe carpal wedging was present in 38% SHOX-noGH, 40% SHOX-GH and 23% TS. Similarly, radial and ulnar lengths were not significantly different between TS and SHOX-D.[br][bold]After 2-yr:[/bold] No significant difference was observed between SHOX-GH and SHOX-noGH for prevalence/severity of any assessed anomaly. However, there were differences at 2-yr between SHOX-GH and TS. Prevalence of radial bowing was greater in SHOX-GH than TS (67% vs 21%; p=0.002). Despite no difference in ht gain, mean[plusmn]SD radial and ulnar lengths were significantly shorter in SHOX-GH vs TS (14.8[plusmn]2.1 vs 16.1[plusmn]1.9 [amp] 16.2[plusmn]2.2 vs 17.6[plusmn]2.1 cm respectively; p=0.03 for both). Carpal wedging was more severe in SHOX-GH than TS (moderate/severe 41% vs 8%; p=0.01).[br][bold]Conclusions:[/bold][br]Radiographic findings of tibial tuberosities and Kosowicz sign are suggestive of TS. Presence of forearm anomalies such as radial bowing and carpal wedging are suggestive of SHOX-D, but also occur in TS. After 2-yr, there were no significant differences between SHOX-GH and SHOX-noGH in any assessed radiological anomalies, indicating that 2-yr GH treatment had no significant effect on skeletal anomalies in SHOX-D.[br][br]Sources of Research Support: Funded by Eli Lilly and Company.[br][br]Disclosures: CJC: Employee, Eli Lilly & Company. CJ: Employee, Eli Lilly & Company. JLR: Scientific Board Member, Eli Lilly & Company; Speaker, Eli Lilly & Company. GAR: Research Funding, Eli Lilly & Company; Speaker, Eli Lilly & Company; Collaborator, Eli Lilly & Company. CAQ: Employee, Eli Lilly & Company. AGZ: Employee, Eli Lilly & Company. WFB: Employee, Eli Lilly & Company. Nothing to Disclose: GK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 390 318 2499 MON-626 PO30-02 Monday 2168 2012


2164 ENDO12L_MON-627 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Final Height in Czech Patiens with Turner Syndrome Following Growth Hormone Therapy Jirina Zapletalova, Darina Aleksijevic, Jana Cerna, Jindrich Cizek, Vlasta Janstova, Bozena Kalvachova, Jarmila Klabochova, Stanislava Kolouskova, Lidka Lisa, Olga Magnova, Dana Novotna, Renata Pomahacova, Jaroslav Skvor, Marta Snajderova, Zdenek Sumnik, Jan Lebl Faculty of Medicine, Palacky University, Olomouc, Czech Republic; University Hospital, Ostrava, Czech Republic; Hospital Cesk[eacute] Budejovice, Cesk[eacute] Budejovice, Czech Republic; Institute of Endocrinology, Prague, Czech Republic; Hospital sv Jir[iacute], Plzen, Czech Republic; Second Faculty of Medicine, Charles University, Prague, Czech Republic; Masaryk University, Brno, Czech Republic; Faculty of Medicine, Charles University, Plzen, Czech Republic; Masaryk Hospital, Usti nad Labem, Czech Republic [bold]Objective: [/bold]Recombinant human growth hormone (GH) therapy in patients with Turner syndrome (TS) was approved for routine use in the Czech Republic in 1992. Although the positive impact of GH administration on final height (FH) has been well established, results of several studies have been controversial in describing height gain between 3 and 15 cm.[br][bold]Aim of the study: [/bold]To evaluate FH in Czech patiens with TS after GH therapy and to identify factors affecting their FH.[br][bold]Subjects and methods: [/bold]A total of 165 patients with TS (karyotype 45,X in 42%) were receiving GH in daily dose 0.05 mg/kg for 3.9 (median; range 1.9-12.4) years up to final height. Median of age of TS diagnosis was 10.0 (0.1-14.4) yrs and age at start of GH treatment 12.1 (3.5-16.5) yrs. Puberty started spontaneously in 22.5% patiens (at age of 12.0; range 8.9-16.0 yrs), Estrogen treatment was initiated in 77.5% girls at 13.9 (8.5-14.8) yrs and the therapy was managed by a pediatric endocrinologists or by a gynecologists. Projected final height was established using the TS growth standards according to Ranke.[br][bold]Results: [/bold]The median of FH was 155.0 (137.5 -168.8) cm, +9.0 (-8.5 - +28.8) cm above projected height. Girls with spontaneous puberty were shorter (153.0; range 143.0-162.8) cm than those with induced puberty (156.0; range 137.5-168.8) cm (p= 0.05). FH of patients treated with estrogens by an endocrinologist (n=65) was higher than that of patients treated by a gynecologist (156.5; range 149.5-168.8 vs. 151.8; range 137.5 - 161.9) cm (p[lt] 0.001). Statistical analysis revealed that age at GH start (p= 0.02) and number of years of GH therapy before spontaneous or induced puberty (p= 0.0049) were the strongest predictors of height gain.[br][bold]Conclusion[/bold]: Early diagnosis of TS allows starting GH therapy at a younger age and administering GH for sufficient time before the onset of estrogen substitution or spontaneous puberty to achieve an optimal FH.[br][br]Nothing to Disclose: JZ, DA, JC, JC, VJ, BK, JK, SK, LL, OM, DN, RP, JS, MS, ZS , JL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1091 318 2500 MON-627 PO30-02 Monday 2169 2012


2165 ENDO12L_MON-628 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Comparative Response to Growth Hormone Therapy in Growth Hormone[ndash]Naive Patients with Noonan Syndrome and Turner Syndrome: Analysis of 4-Year Data from the ANSWER Program[reg] Peter A Lee, Judith L Ross, Robert Z Gut, Vatsala Karwe, John A Germak Pennsylvania State College of Medicine, Hershey, PA; Thomas Jefferson University and DuPont Hospital for Children, Philadelphia; Wilmington, PA; Novo Nordisk Inc, Princeton, NJ [bold]Background and Objective: [/bold]Few data exist comparing the growth response to growth hormone (GH) treatment over time between 2 syndromes with similar phenotypes: Noonan syndrome (NS) and Turner syndrome (TS). The present analysis compares changes in height standard deviation score ([Delta]HSDS) and corrected HSDS (HSDS at each time point[minus]target height SDS) at onset of therapy and through 4 years of treatment with Norditropin[sup][reg][/sup] (somatropin rDNA origin, Novo Nordisk A/S, Bagsvaerd, Denmark) in patients with NS and TS enrolled in the American Norditropin Studies: Web-enabled Research (ANSWER) Program[sup][reg][/sup], a US-based registry.[br][bold]Method and Results: [/bold]As of October 2011, 120 patients with NS (90 males, 30 females) and 472 females with TS were enrolled in the ANSWER Program[sup][reg][/sup]. The mean ([plusmn]SD) age of NS subjects was 9.2[plusmn]3.8 years, compared with 8.6[plusmn]4.1 years for TS subjects. Mean ([plusmn]SD) GH dose (mcg/kg/d) at onset of therapy and y4 was 46.5[plusmn]11.3 and 58.6[plusmn]15.7 for NS patients and 51.1[plusmn]9.9 and 52.7[plusmn]12.7 mcg/kg/d for TS patients. The mean ([plusmn]SD) HSDS at onset of therapy for NS and TS, respectively, were [ndash]2.6[plusmn]0.7 and [ndash]2.6[plusmn]0.9. Mean ([plusmn]SD) HSDS continuously increased over 4 years in NS patients (y1: [minus]2.3, n=76; y2: [minus]2.0, n=49; y3: [minus]1.7, n=31; y4: [minus]1.3, n=17). After an initial increase in TS patients, mean HSDS did not change over time (y1: [minus]2.1, n=320; y2: [minus]1.8, n=255; y3: [minus]1.6, n=189; y4: [minus]1.7, n=128). Mean corrected HSDS improved over 4 years in NS subjects (y1: [minus]2.0, n=64; y2: [minus]1.5, n=42; y3: [minus]1.5, n=29; y4, [minus]1.0, n=14) in contrast to TS subjects, who showed a plateau in corrected HSDS after y2 (y1: [minus]2.1, n=265; y2: [minus]1.9, n=218; y3: [minus]1.8, n=161; y4: [minus]1.8, n=110). A negative correlation between [Delta]HSDS and baseline age was observed over the first 2 years of GH treatment (y1, R= [minus]0.316, [italic]P[/italic]=0.006; y2, R= [minus]0.339, [italic]P[/italic]=0.017) for NS subjects; for TS subjects a negative correlation between [Delta]HSDS and age was present only at y1 (R= [minus]0.266, [italic]P[/italic][lt]0.0001).[br][bold]Conclusions: [/bold]NS subjects showed increases in mean [Delta]HSDS and corrected target HSDS over 4 years of GH treatment, in contrast with TS subjects, in which further incremental increases appeared to plateau after y2. Differences in GH responsiveness in NS and TS patients may be multifactorial, potentially involving the GH dose, use of estrogen therapy in TS, and the skeletal dysplasia, related to SHOX deficiency in TS.[br][br]Disclosures: PAL: Advisory Group Member, Abbott Laboratories, Novo Nordisk; Clinical Researcher, Abbott Laboratories, Eli Lilly & Company, Pfizer, Inc., Ipsen. JLR: Advisory Group Member, Novo Nordisk, Eli Lilly & Company, Abbott Laboratories; Clinical Researcher, Novo Nordisk, Pfizer, Inc., Eli Lilly & Company. RZG: Employee, Novo Nordisk. VK: Employee, Novo Nordisk. JAG: Employee, Novo Nordisk. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2302 318 2501 MON-628 PO30-02 Monday 2170 2012


2166 ENDO12L_MON-629 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) GH Treatment in Children with Prader-Willi Syndrome: 3 Years[apos] Longitudinal Data in Prepubertal Children from KIGS Database Anita Hokken-Koelega, Maithe Tauber, Hartmut Wollmann Dutch Growth Research Foundation, Rotterdam, Netherlands; Erasmus University Medical Center Rotterdam/Sophia Children[apos]s Hospital, Rotterdam, Netherlands; H[ocirc]pital des Enfants, Toulouse, France; Pfizer Endocrine Care, Walton Oaks, UK [bold]Background:[/bold][br]Prader Willi Syndrome is a rare disorder due to lack of expression of the paternally derived chromosome 15q11-13, in most children due to a deletion or uniparental maternal disomy. Children with PWS have impaired growth and growth hormone (GH) treatment has shown to improve growth. However, longer term data of a large group of GH-treated prepubertal children with PWS have not yet been reported.[br][bold]Objective:[/bold][br]To evaluate growth during 3 years of GH treatment in a large group of prepubertal children with PWS.[br][bold]Design/Patients:[/bold][br]415 prepubertal children (224 boys) with PWS who were treated with GH for 3 years. Their longitudinal data were registered in KIGS database (Pfizer International Growth Database).[br][bold]Results:[/bold][br]Mean(SD) birth weight and birth length SD score (SDS) were -1.25 (1.1) and -0.23 (1.4), resp. Mean (SD) mid parental height SDS was -0.1 (1.1). Prior to start of GH, 128 children had a GH stimulation test with a mean (SD) GH peak of 8,1 (9,3) ug/l and mean (SD) serum IGF-I SDS -1.1 (1.5). All children remained prepubertal during the 3 years of GH treatment. Mean (SD) GH dose was 0.23 (0.06) mg/kg/wk.[br]At baseline and after 1, 2 and 3 years of GH treatment the following mean (SD) values were found: Age (yr), 4.5 (2.9), 5.5 (2.9), 6.5 (2.9) and 7.5 (2.9), resp.; Height SDS (Prader reference (Helv. Paed. Acta 1988)), -2.1 (1.4), -1.1 (1.4), -0.7 (1.4), and -0.4 (1.4), resp.; H-MPHH SDS, -2.0 (1.6), -1.0 (1.5), -0.6 (1.5), and -0.3 (1.4), resp.; HV (cm/yr), 6.6 (3.5), 11.1 (2.9), 8.3 (2.0), and 7.3 (1.8), resp.; Weight SDS, -0.2 (2.1), 0.3 (1.8), 0.7 (1.6), and 1.0 (1.5), resp.; Bone age (yr), 4.3 (2.9), 5.6 (3.4), 7.0 (3.6), and 8.5 (3.5), resp.[br][bold]Conclusions:[/bold][br]These data from a very large group of prepubertal children with PWS demonstrates that growth hormone treatment significantly improves growth of these children, resulting in a complete normalization of their stature within a few years.[br][br]Nothing to Disclose: AH-K, MT, HW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1476 318 2502 MON-629 PO30-02 Monday 2171 2012


2167 ENDO12L_MON-630 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) The Proportion of Uniparental Disomy Is Increased in Prader-Willi Syndrome Due to an Advanced Maternal Childbearing Age in Korea Se Hyun Maeng, Doosoo Kim, You Jin Jung, Sung Yoon Cho, Chang-Seok Ki, Young Bae Sohn, Su Jin Kim, Dong-Kyu Jin Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea; Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea; Ajou University Hospital, Suwon, Republic of Korea; National Cancer Center, Goyang, Republic of Korea Prader-Willi syndrome (PWS) is caused by paternal interstitial deletion of the 15q11[ndash]13 region or maternal uniparental disomy (mUPD). Four possible mechanisms can give rise to mUPD: trisomy rescue (TR), gamete complementation (GC), monosomy rescue (MR), and postfertilization mitotic nondisjunction (Mit). TR/GC is caused by nondisjunction at maternal meiosis 1[M1] or 2[M2], and [M1] errors have been correlated with increased maternal age. We examined the effects of maternal age on the distribution of genotypes in PWS by comparing the maternal age of PWS patients at birth according to their genotypes.[br]We classified 97 Korean PWS patients whose parental age was available. Deletions were found in 66 patients, [M1]TR/GC in 15 patients, [M2]TR/GC in 2 patients, MR/Mit in 4 patients, and epimutation in 2 patients. Maternal ages at birth showed a significant difference between the deletion group [median age of 29, IQR=(27,31)] and [M1]TR/GC group [median age of 35, IQR=(31,38)] (P [lt] 0.0002). The relative birth frequency of the [M1] TR/GC group was much increased by since 2006 compared to until 2005. In summary, we report an increasing tendency for [M1]TR/GC due to advanced maternal childbearing age in Korea.[br][br]Nothing to Disclose: SHM, DK, YJJ, SYC, C-SK, YBS, SJK, D-KJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 399 318 2503 MON-630 PO30-02 Monday 2172 2012


2168 ENDO12L_MON-631 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Spectrum of the [italic]AAAS[/italic] Gene Mutations in Japanese Patients with Allgrove Syndrome Noriyuki Katsumata, Satoru Ikemoto Natl Res Inst for Child Hlth and Dev, Tokyo, Japan; Jikei Univ Aoto Hosp, Tokyo, Japan [bold]Background:[/bold] Allgrove syndrome or triple A syndrome is a rare autosomal recessive disorder, characterized by adrenal insufficiency due to ACTH resistance, alacrima, and achalasia. This syndrome is caused by mutations in the [italic]AAAS[/italic] gene, which encodes a protein of 546 amino acids, ALADIN. In the present study, we analyzed the [italic]AAAS[/italic] gene in Japanese patients with Allgrove syndrome to elucidate its mutational spectrum in Japan.[br][bold]Subjects:[/bold] Four unrelated Japanese patients with Allgrove syndrome were studied. All of them were born to consanguineous parents after uncomplicated full-term gestations, and developed glucocorticoid insufficiency at 2 to 5 years of age and alacrima before 8 years of age. Two of them were diagnosed as having achalasia at 2 and 3 years of age, and the remaining two had not developed achalasia.[br][bold]Genetic analysis: [/bold]The genetic analysis was performed after written informed consent was obtained from the parents. Sequencing analysis of the [italic]AAAS[/italic] gene revealed that each patient was homozygous for a p.R119X, p.R194X, p.Q237X or IVS7+1G[gt]A mutation.[br][bold]Discussion:[/bold] The p.R119X, p.R194X and p.Q237X mutations are reported not only in Japanese patients with Allgrove syndrome in a homozygous state, but also in those of the other ethnic groups in a compound heterozygous state. Therefore, these are rare but recurrent mutations. The IVS7+1G[gt]A mutation has not been previously reported, thus this is a novel mutation. The IVS7+1G[gt]A mutation destroys the highly conserved consensus splice donor site sequence and presumably causes aberrant splicing of [italic]AAAS[/italic] mRNA to give rise to the syndrome.[br][bold]Conclusion: [/bold]1) We have identified a novel [italic]AAAS[/italic] gene mutation, IVS7+1G[gt]A in a Japanese patient with Allgrove syndrome. 2) There are no mutational hot spots in the [italic]AAAS[/italic] gene in Japanese patients with Allgrove syndrome, and the mutations are unique to the respective families.[br][br]Nothing to Disclose: NK, SI 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 252 318 2504 MON-631 PO30-02 Monday 2173 2012


2169 ENDO12L_MON-632 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Predictors of Hyponatremia and Hypopituitarism in Patients with Diabetes Insipidus at Seattle Children[apos]s Hospital David M Werny, Clinton Elfers, Faisal Malik, Cate Pihoker, Christian Roth Seattle Children[apos]s Hospital, Seattle, WA; Seattle Children[apos]s Research Institute, Seattle, WA Central Diabetes Insidpidus (CDI) is a rare but potentially life threatening disorder of osmoregulation. CDI is commonly treated with DDAVP, one adverse effect of which is hyponatremia. Some anti-epileptics may lower sodium levels (1), potentially increasing the risk for CDI patients already at higher risk of seizure. Patients with ACTH deficiency may also be more susceptible to hyponatremia (2). Here we describe the creation of a database of CDI patients seen at Seattle Children[apos]s Hospital (SCH), as well as analyses on predictors of hyponatremia and anterior pituitary deficiencies (APDs) in our population of CDI patients.[br]We are retrospectively analyzing all charts for patients seen in the SCH outpatient health system with a diagnosis of CDI. Patients were divided by their CDI cause: anatomic, infiltrative, and genetic/idiopathic. A [ldquo]hyponatremic episode[rdquo] was defined as any outpatient encounter where the sodium was [lt] 135, or any hospitalization that contained a sodium [lt] 135.[br]As of January 2012 we have identified 51 patients with CDI in the SCH system. Etiologies (with % in parenthesis) included: anatomic (29%), infiltrative (53%), and genetic/idiopathic (18%). 75% had anterior pituitary deficiencies (APDs), and 59% had ACTH deficiency. Those with an anatomic or infiltrative etiology were more likely to have APDs (p [lt] 0.01) and episodes of hyponatremia (1.9 vs 0.2, p = 0.04). There was no association between having a history of seizure and having episodes of hyponatremia (p = 0.16), but few patients had a history of seizure (n = 12). A positive association between ACTH deficiency and hyponatremic episodes was not statistically significant (2.1 vs 0.86, p = 0.10).[br]We estimate that our retrospective review will find roughly 100 children with CDI at SCH. Those with anatomic or infiltrative etiologies are more likely to have APDs, possibly because of the greater likelihood of surgical intervention. They are also more likely to have episodes of hyponatremia which may again be related to the increased complexity of care that anatomic and infiltrative CDI can require relative to other etiologies. What co-morbidities, medications, and procedures within this population that result in hyponatremia will be the subject of future investigations. Those with ACTH deficiency or a history of seizures may be hyponatremic more often. With a larger sample size we can clarify the risks for these potentially vulnerable subgroups of CDI patients.[br][br](1) Toumba M, Stanhope R. Morbidity and Mortality Associated with Vasopressin Analogue Treatment. Journal of Pediatric Endocrinology and Metabolism. 2006. 19: 197-201. (2) Rizzo V et al. Morbidity and mortality associated with vasopressin replacement therapy in children. Journal of Pediatric Endocrinology and Metabolism. 2001. 14: 861-867.[br][br]Nothing to Disclose: DMW, CE, FM, CP, CR 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 621 318 2505 MON-632 PO30-02 Monday 2174 2012


2170 ENDO12L_MON-633 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Can Inhibin B Be Utilized as a Marker of Seminiferous Tubule Impairment in Children with Cryptorchidism? Daniel P Casella, Jeffrey J Tomaszewski, Peter A Lee, Francis X Schneck, Selma F Witchel Children[apos]s Hospital of University of Pittsburgh Medical Center, Pittsburgh, PA; Pennsylvania State College of Medicine, Hershey, PA [bold]Background and Objective: [/bold]Seminiferous tubule dysfunction resulting in infertility is a common complication of cryptorchidism. A direct hormonal bioassay of seminiferous tubule function would be useful in diagnosis and management of boys with conditions that involve the testis. Inhibin B is known to be an effective marker of spermatogenesis in adults [1-4], however several studies have reported no difference in inhibin levels in children with cryptorchidism compared to controls [5-7]. The role of the postnatal FSH surge and its relationship to inhibin B levels is also controversial with studies reporting negative, positive and no correlation in control and cyrptorchid populations. [5, 8-10].[br][bold]Objective:[/bold] To examine the association between serum inhibin B levels and age in children with cryptorchidism undergoing orchiopexy.[br][bold]Methods:[/bold] 81 boys from 5 to 26 months of age undergoing elective orchiopexy for cryptorchid testis/es were prospectively enrolled. Serum inhibin B, FSH, and LH levels were obtained at the time of surgery immediately following the induction of anesthesia. Inhibin B levels were measured by a double-antibody enzyme-linked immunosorbent assay from Serotec Ltd. (Oxford, UK). Results were stratified by age and compared to published controls.[10].[br][bold]Results:[/bold] Within the 4.5-[lt]7.5, 7.5-[lt]10.5 and 10.5-[lt]13.5 month age groups, mean inhibin B levels were significantly decreased in cryptorchid patients (234.2 pg/ml vs. 351 pg/ml, p[lt]0.02; 201.6 vs. 281pg/ml, p[lt]0.02; 143.6 vs 235pg/ml, P[lt]0.001). There was no difference in mean inhibin B levels in the 13.5-[lt]17.5, 17.5-[lt]19.5, 19.5[lt]22.5 and 22.5-[lt]26.5 month age groups. We also observed no significant correlation between FSH and inhibin B in the overall cohort of cryptorchid patients.[br][bold]Conclusions:[/bold] When compared to published controls, cryptorchid infants between 5 and 13.4 months of age had significantly lower inhibin B levels. Inhibin B levels were not correlated with FSH levels, suggesting that they are reflective of overall sertoli cell mass as opposed to representing response to the postnatal FSH/LH surge. Long term follow-up will be required to determine whether early inhibin B levels are predictive of post-pubertal seminiferous tubule function in children with cryptorchidism.[br][br]1. Anawalt, B.D., et al., Serum inhibin B levels reflect Sertoli cell function in normal men and men with testicular dysfunction. J Clin Endocrinol Metab, 1996. 81(9): p. 3341-5. 2. Myers, G.M., G.M. Lambert-Messerlian, and M. Sigman, Inhibin B reference data for fertile and infertile men in Northeast America. Fertil Steril, 2009. 92(6): p. 1920-3. 3. Chatzidarellis, E., et al., Effects of taxane-based chemotherapy on inhibin B and gonadotropins as biomarkers of spermatogenesis. Fertil Steril, 2010. 94(2): p. 558-63. 4. Laporte, S., et al., Inhibin B and anti-Mullerian hormone as markers of gonadal function after hematopoietic cell transplantation during childhood. BMC Pediatr, 2011. 11: p. 20. 5. Barthold, J.S., et al., Reproductive Hormone Levels in Infants with Cryptorchidism during Postnatal Activation of the Pituitary-Testicular Axis. The Journal of Urology, 2004. 172(4): p. 1736-1741. 6. Suomi, A.M., et al., Hormonal changes in 3-month-old cryptorchid boys. J Clin Endocrinol Metab, 2006. 91(3): p. 953-8. 7. Pierik, F.H., et al., The hypothalamus-pituitary-testis axis in boys during the first six months of life: a comparison of cryptorchidism and hypospadias cases with controls. Int J Androl, 2009. 32(5): p. 453-61. 8. Crofton, P.M., et al., Inhibin B in boys from birth to adulthood: relationship with age, pubertal stage, FSH and testosterone. Clin Endocrinol (Oxf), 2002. 56(2): p. 215-21. 9. Andersson, A.M., et al., Serum inhibin B in healthy pubertal and adolescent boys: relation to age, stage of puberty, and follicle-stimulating hormone, luteinizing hormone, testosterone, and estradiol levels. J Clin Endocrinol Metab, 1997. 82(12): p. 3976-81. 10. Andersson, A.M., et al., Longitudinal reproductive hormone profiles in infants: peak of inhibin B levels in infant boys exceeds levels in adult men. J Clin Endocrinol Metab, 1998. 83(2): p. 675-81.[br][br]Nothing to Disclose: DPC, JJT, PAL, FXS, SFW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 256 318 2506 MON-633 PO30-02 Monday 2175 2012


2171 ENDO12L_MON-634 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Penile and Clitoral Sizes of Normal Full-Term Newborns in Korea So Eun Park, Sung Yeon Ahn, Choong Ho Shin College of Medicine, CHA University, Seoul, Korea; Kangwon National University Hospital, Chuncheon, Korea; College of Medicine, Seoul National University, Seoul, Korea Background:[br]Abnormal genital size is an early signs of underlying endocrine diseases and previous reports show ethnic differences in normal length.[br]Objective:[br]We studied to establish reference range of genital size in normal full term in Korea and compare that with other ethnicities. We present the largest study of penile and clitoral length measurement from Korea.[br]Subjects:[br]Normal full term males (n=440) and females (n=390) with birth weight appropriate for gestational age were included.[br]Methods:[br]Penile length was measured with a rigid ruler as the distance from the pubic bone to the tip of glans or stretched flaccid length. Clitoral length and width were measured with a tumorimeter in after the labia major were spread in frog-leg position.[br]Measurements were taken 3 times by a single examiner during the first 3 days of life. The study was carried out in Gangnam CHA medical center, CHA University over October 2011 to December 2011.[br]Results:[br]The mean penile length was 2.7cm (standard deviation(SD) 0.3). The mean clitoral width and length were 6.1 mm (SD 1.4) and 3.4 mm (SD 0.9) respectively.[br]Conclusion:[br]This study establishes reference range for genital size of normal full term newborns in Korea which would help clinicians to diagnose early and treat an underlying diseases promptly.[br][br]Nothing to Disclose: SEP, SYA, CHS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1220 318 2507 MON-634 PO30-02 Monday 2176 2012


2172 ENDO12L_MON-635 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) The Correlation of Serum Insulin-Like Growth Factor Binding Protein-3 (IGFBP-3) with Glycemic Control and Lipid Metabolism in Adolescents with Type 2 Diabetes Min Sun Kim, Young Hwa Kong, Pyoung Han Hwang, Dae-Yeol Lee Chonbuk National University Medical School, Jeonju, Republic of Korea [bold]BACKGROUND[/bold]: The aim of this study was to investigate the hypothesis that serum insulin-like growth factor-1(IGF-I) and IGFBP-3 levels are possible markers for glycemic control, lipid metabolism and insulin resistance, and they are correlated with clinical and laboratory parameters in adolescents with type 2 diabetes. [bold]METHODS:[/bold] We included 66 adolescents aged 10 to 18 years (42 girls and 24 boys) with asymptomatic glucosuria detected by school urine screening. They were classified into 3 groups according to the results of oral glucose tolerance test (OGTT); group 1, normal glucose tolerance group, n=32; group 2, impaired glucose tolerance group, n=11; group 3, diabetes group, n=23. We performed anthropometric measurement and laboratory tests including fasting lipid profile (cholesterol, triglyceride and LDL cholesterol), fasting plasma glucose (FPG), insulin, HbA1c, c-peptide, IGF-I, IGFBP-3 and HOMA-IR (homeostasis model of assessment-insulin resistance). [bold]RESULTS[/bold]: 1) Serum IGF-I and IGFBP-3 levels were significantly higher in group 3(467.5[plusmn]159.5 ng/mL and 6938.4[plusmn]1232.9 ng/mL, respectively), than in group 1(351.6[plusmn]119.1 ng/mL and 5230.0[plusmn]909.2 ng/mL, p[lt]0.001) and group 2(391.7[plusmn]94.5 ng/mL and 5756.0[plusmn]794.6 ng/mL). Serum IGF-I and IGFBP-3 had positive correlation with FPG, HbA1c, c-peptide, and HOMA-IR. Also serum IGFBP-3 level showed the positive correlation with BMI and lipid profile. 2) In group 1, serum IGF-I level was correlated with only serum IGFBP-3. Serum IGFBP-3 has positively associated with age and HbA1c. And obese normal subjects(n=8, 25.8%) showed significantly higher serum c-peptide, insulin, cholesterol, LDL and HOMA-IR than those of non-obese normal subjects(n=23, 74.2%). 3) In group 3, serum IGF-I level showed positive correlation with HbA1c and FPG, OGTT glucose levels. Serum IGFBP-3 level had positive correlation with FPG, HbA1c, c-peptide and lipid profile. However, there was only significant difference in c-peptide level between obese diabetic subjects(n=17, 77.3%) and non-obese diabetic subjects(n=5, 22.7%). [bold]CONCLUSION[/bold]: Serum IGF-I and IGFBP-3 levels were significantly elevated in adolescents with type 2 diabetes. They were positively correlated with HbA1c and FPG, and serum IGFBP-3 level was significantly associated with lipid profile, irrespective of obesity. These findings suggest that serum IGFBP-3 may use as a marker for glycemic control and/or development of dyslipidemia in adolescents with type 2 diabetes.[br][br]Nothing to Disclose: MSK, YHK, PHH, D-YL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1383 318 2508 MON-635 PO30-02 Monday 2177 2012


2173 ENDO12L_MON-636 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Low Level of 25-Hydroxyvitamin D in Korean Children and Adolescents with Type 1 Diabetes Mellitus Hyo-Kyoung Nam, Eun He Cho, Young Jun Rhie, Kee-Hyoung Lee College of Medicine, Korea University, Seoul, Republic of Korea [bold]Objectives: [/bold]Vitamin D is needed for insulin secretion. A low vitamin D level has been associated with increased risk of diabetes mellitus (DM), but the association has not been evaluated in Asian children and adolescents. The aim of this case-control study was to assess the vitamin D status in youth with type 1 DM and to examine the influence of specific factor on 25-hydroxyvitamin D level in type 1 DM children and adolescents.[br][bold]Patients and Methods: [/bold]The study was carried out among type 1 DM cases (n=85) and healthy controls (n=518) below 20 years at the pediatric endocrinology clinics of Korea university hospital. We recorded auxological data and measured biochemical levels including serum 25-hydroxyvitamin D.[br][bold]Results: [/bold]Mean value of vitamin D was significantly lower in youth with Type 1 DM compared to that of non-diabetics controls (21.5 [plusmn] 8.5 ng/ml vs. 28.0 [plusmn] 12.0 ng/ml, [italic]p =[/italic] 0.001). Of the 603 subjects 177 (29%) were vitamin D deficient, defined as a 25-hydroxyvitamin D level below 20 ng/ml. The study revealed that the prevalence of vitamin D deficiency was considerably higher in Type 1 DM cases (48%) compared to non-diabetic controls (26%). The serum 25-hydroxyvitamin D level was negatively correlation with age(r = -0.317, [italic]p [/italic]= 0.001) and BMI(r = -0.143, [italic]p[/italic] = 0.001). However, duration of treatment, HbA1c, total unit of insulin and insulin dose per body weight were not correlated with 25-hydroxyvitamin D level in youth with type 1 DM.[br][bold]Conclusions: [/bold]Prevalence of vitamin D deficiency in type 1 diabetic children and adolescents is high. Further studies investigating the mechanisms leading to 25-hydroxyvitamin D deficiency in children and adolescents with type 1 diabetes will be needed.[br][br]Nothing to Disclose: H-KN, EHC, YJR, K-HL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 387 318 2509 MON-636 PO30-02 Monday 2178 2012


2174 ENDO12L_MON-637 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Variation in Resource Utilization and Outcomes for Diabetic Ketoacidosis in Children Faisal S Malik, Rajendu Srivastava, Lisa McLeod, Ron Keren, Xianqun Luan, Russell Localio, Samir S Shah, Karen M Wilson, Joel S Tieder Seattle Children[apos]s Hospital/University of Washington, Seattle, WA; Primary Children[apos]s Medical Center, Salt Lake City, UT; Children[apos]s Hospital of Philadelphia, Philadelphia, PA; University of Pennsylvania School of Medicine, Philadelphia, PA; Cincinnati Children[apos]s Hospital Medical Center, Cincinnati, OH; Children[apos]s Hospital, Aurora, CO Background: Diabetic ketoacidosis (DKA) is one of the most serious and potentially preventable conditions affecting hospitalized children. Our goal was to identify opportunities to improve care by 1) examining the variation in resource utilization, cost and clinical outcomes for the management of DKA across major U.S. children[apos]s hospitals and 2) explore the association between resource utilization and clinical outcomes.[br]Methods: Multi-center retrospective cohort study of children 2-18 years of age admitted to 38 freestanding children[apos]s hospitals participating in the Pediatric Health Information Systems (PHIS) database between January 2004 and December 2009 with a discharge diagnosis code for DKA. We applied a standardized costing method to calculate resource utilization per hospitalization. Using a mixed effects model, we determined risk-adjusted differences in observed versus expected resource utilization for the mean total standardized costs, classifiable clinical cost categories (e.g. pharmacy), and readmission within 30 and 365 days. A multivariate analysis adjusting for patient characteristics tested the association of length of stay (LOS) with readmission risk.[br]Results: The study included 24,890 children. Across hospitals there was wide variability in mean standardized hospital costs ($7,195; min $4,144, max $14,479). The clinical resources that contributed most to the mean standardized hospital cost were LOS (55.9%), followed by laboratory (20.1%) and pharmacy (8.3%). There were 17 in-hospital deaths and the mean rate of cerebral edema was 2.7% (0.6, 20.7). Readmission at one year was common and variable (18.7%; 6.5, 41.1). Across-hospital variability in resource use and readmission persisted after adjusting for covariates. Patients with a longer non-ICU length of stay had a significantly lower risk-adjusted odds of readmission at 30 days (OR 0.91, p = 0.02) and 365 days (OR 0.94, p = 0.005), regardless of total and ICU lengths of stay.[br]Conclusion: Hospital readmission for DKA is extremely common. Even after adjusting for patient characteristics, there is significant variation in resource utilization and readmission for children with DKA. Successful management strategies, such as longer non-ICU admissions, should be explored further to inform improvement efforts and comparative effectiveness studies aimed at improving the quality of DKA care in the US.[br][br]Sources of Research Support: Grant from the Child Health Corporation of America.[br][br]Nothing to Disclose: FSM, RS, LM, RK, XL, RL, SSS, KMW, JST 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1831 318 2510 MON-637 PO30-02 Monday 2179 2012


2175 ENDO12L_MON-638 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Reversal of Fortune: The Development of Diabetes following Surgery to Treat Congenital Hyperinsulinism Katherine Lord, Heather McKnight-Menci, Diva D De Leon The Children[apos]s Hospital of Philadelphia, Philadelphia, PA Background: Inactivating mutations of the ATP sensitive potassium (K[sub]ATP[/sub]) channel cause the most common and severe form of congenital hyperinsulinism (HI). Most children with K[sub]ATP[/sub]HI require a pancreatectomy to control the hypoglycemia. Limited resection is curative for children with the focal form of the disease, but children with diffuse K[sub]ATP[/sub]HI require near-total pancreatectomy. This approach places children at risk of glucose intolerance and diabetes.[br]Objective: To describe the development of diabetes following pancreatectomy in children with hyperinsulinism.[br]Methods: Medical records were reviewed and patient interviews were conducted.[br]Results: Twelve children (7F, 6M) underwent near-total pancreatectomy (90-98% resection) at a median age of 34 days (6-70 days). Three subjects required additional pancreatic resection for continued hypoglycemia. In all cases, histology showed nucleomegaly throughout the pancreas, consistent with diffuse HI. Ten subjects were homozygous or compound heterozygous for mutations in [italic]ABCC8[/italic], one had no mutations identified in the four most common HI genes and one was not tested. Four out of 12 developed diabetes in the immediate post-operative period and were discharged on insulin. The other eight subjects developed diabetes at a median age of 12.2 years (5.3-20.8 years) with a mean hemoglobin A1c of 7.4% (6.7-8.1%) at diagnosis. Diagnosis of diabetes was based on either an elevated fasting glucose or hemoglobin A1c. Subjects denied symptoms of diabetes. Of these eight subjects, five were started on insulin at the time of diagnosis. Of two subjects initially treated with metformin, one required insulin 5 years after diagnosis and the other remains only on metformin. One subject was initially diet controlled, but required insulin within 4 months of diagnosis. The median duration of diabetes is 6.9 years (0.8-21.6 years) and their mean insulin requirement is 0.5 units/kg/day (0.1-0.7 units/kg/day).[br]Conclusions: The development of diabetes after pancreatectomy in children with hyperinsulinism is highly variable, with some children presenting in the post-operative period and others as late as 20 years after their surgery. Although mild at presentation, most cases progress and eventually require insulin. However, insulin requirements are lower than expected for the duration of diabetes, suggesting there is residual beta cell function.[br][br]Nothing to Disclose: KL, HM-M, DDDL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 823 318 2511 MON-638 PO30-02 Monday 2180 2012


2176 ENDO12L_MON-639 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Hemoglobin A1c and Glucose Intolerance in Obese Children and Adolescents You Jin Kim, Hae Sang Lee, Hong Kyu Park, Jung Hee Ko, Jin Soon Hwang Ajou University School of Medicine, Ajou University Hospital, Suwon, Korea [bold]Aims: [/bold]Childhood obesity is associated with an increased likelihood for having impaired glucose tolerance (IGT), dyslipidemia, and diabetes. The goal of study was to evaluate glycated hemoglobin A1c (HbA1c) as a screening test for IGT in obese children and adolescents and identify the optimal HbA1c threshold.[br][bold]Methods: [/bold]We studied 126 obese and overweight children (body mass index [gt]85[sup]th[/sup] percentile for age and gender) 4-17 years of age referred to the endocrine clinic at Ajou University Hospital in Korea. All subjects underwent HbA1c and oral glucose tolerance test.[br][bold]Results:[/bold] Thirty-four patients (27%) out of 126 had IGT. Silent diabetes was diagnosed in 10 adolescents (7.9%). Based on the ROC curve, the optimal cut point of HbA1c related to IGT diagnosed by OGTT was 40 mmol/mol (5.8%), which was associated with an 64.7% sensitivity and 61.6% specificity, with an area under the ROC curve of 0.651 (95% confidence interval 0.529-0.772).[br][bold]Conclusion: [/bold]Obesity associated with an increased risk of IGT. A HbA1c value of 40 mmol/mol (5.8%) should be used as a screening tool to identify children and adolescents with IGT.[br][br]Nothing to Disclose: YJK, HSL, HKP, JHK, JSH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 517 318 2512 MON-639 PO30-02 Monday 2181 2012


2177 ENDO12L_MON-640 POSTER SESSION: Pediatric Endocrinology [amp] Cancer (1:30 PM-3:30 PM) Serum Alkaline Phosphatase Levels in Healthy Korean Children and Adolescents of 10[ndash]20 Years [mdash] Based on Korea National Health and Nutrition Examination Surveys 2009 Ji Young Seo, Jung Sub Lim Eulji Hospital, Eulji University, Seoul, Republic of Korea; Korean Cancer Center Hospital, Seoul, Republic of Korea [bold]PURPOSE[/bold]: Serum alkaline phosphatase (sALP) levels show great variation with age and sex in children and adolescents. We aimed to study pediatric age- and sex-specific reference ranges for sALP and its change pattern according to age.[br][bold]METHODS[/bold]: Cross sectional results from 1366 healthy Korean children and adolescent (10-19.9 years) in Korean National Health and Nutrition Examination Surveys was analyzed. We made age- and sex- specific reference data for sALP by Hitachi Automatic Analyzer 7600 using Pureauto S ALP. We assessed its peak age and relation to height.[br][bold]RESULTS:[/bold] Reference values for sALP according to age were constructed. Peak sALP levels were noted at 12-13 years in boys (p[lt]0.001). Total sALP levels decreased to the adult level at the age of 17 in boys and 15 in girls, respectively.[br][bold]CONCLUSIONS[/bold]: This study provides reference values for sALP according to age of Korean children and adolescent. This normative data with upper and lower limit will provide a basis for better evaluation of sALP levels especially bone disorders such as rickets and vitamin D deficiency. However, caution should be made in interpretation of sALP levels because different buffers might cause different values in same serum.[br][br]Nothing to Disclose: JYS, JSL 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2380 318 2513 MON-640 PO30-02 Monday 2182 2012


2178 ENDO12L_MON-641 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) The Synthesis and Activity of Inhibin and Activin Are Regulated by Prodomain Interactions Kelly L Walton, Emily K Kelly, David M Robertson, Craig A Harrison Prince Henry[apos]s Institute of Medical Research, Clayton, Australia; Prince Henry[apos]s Institute of Medical Research, Clayton, Australia Inhibin A and activin A reciprocally regulate follicle stimulating hormone synthesis by the pituitary. Similar to other TGF-[beta] proteins, inhibin A and activin A are synthesised as large precursor molecules, comprising N-terminal pro- and C-terminal mature domains. The pro-inhibin heterodimer comprises [alpha]- and [beta][sub]A[/sub]-subunit chains, whereas pro-activin is a homodimer of two [beta][sub]A[/sub] chains. We have previously shown that key residues in an [alpha]-helix ([alpha][sub]1[/sub]) at the N-terminus of the prodomain direct the folding and dimerisation of pro-inhibin A and activin A. In this study, we determined how proteolytic processing of the inhibin and activin precursors influences ligand secretion and bioavailability. The inhibin [alpha]- and [beta][sub]A[/sub]-subunit prodomains are enzymatically cleaved from their mature domains by proprotein convertases (PC) at consensus RXXR sites (site[sub]1[/sub]). The inhibin [alpha]-subunit is unique within the TGF-b superfamily as it encodes a second PC cleavage site (site[sub]2[/sub]) within its prodomain. We expressed inhibin [alpha]- and [beta][sub]A[/sub]-subunits in various mammalian cell lines, and examined the extent of precursor processing by Western blot. Activin A ([beta][sub]A[/sub]-subunit) was processed with comparable efficiencies in all cell lines tested. Interestingly, inhibin A was found to be poorly processed by cos-7 cells, which are known to lack PC5/6. This finding suggests that PC5/6 may be the endogenous enzyme processing the inhibin [alpha]-subunit, although it is not known whether it is required for cleavage at both sites. To determine whether proteolytic processing precedes or follows subunit dimerisation, we silenced the individual cleavage sites in the inhibin [alpha]-subunit using [italic]in vitro [/italic]mutagenesis and examined the effect on biosynthesis. Mutation of site[sub]1[/sub] between the pro- and mature domains had no effect on inhibin production. However, silencing of site[sub]2[/sub] in the inhibin [alpha]-subunit prodomain, abolished the synthesis of inhibin A. It is likely that this cleavage event occurs intracellularly and is required prior to heterodimerisation of the [alpha]- and [beta][sub]A[/sub]-subunits. Cleavage at site[sub]2[/sub] in the inhibin [alpha]-subunit releases a 43-amino acid pro[alpha]-peptide, comprising the important [alpha][sub]1[/sub]-helix. Ligand blot analysis indicated that the pro[alpha]-peptide binds specifically to mature inhibin, and was unable to bind activin. Furthermore, the pro[alpha]-peptide was capable of limiting inhibin A bioactivity in primary rat pituitary cells. These studies signify that inhibin [alpha]-subunit processing limits the synthesis and bioavailability of inhibin A.[br][br]Sources of Research Support: Program Grant 241000 from the National Health and Medical Research Council of Australia, and the Victorian Government[apos]s Operational Infrastructure Support Program.[br][br]Nothing to Disclose: KLW, EKK, DMR, CAH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1373 319 2514 MON-641 PO17-02 Monday 2183 2012


2179 ENDO12L_MON-642 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) FSTL3 Deletion Reveals Roles of TGF[beta] Signaling in Skeletal Development Waheed Mahmood, Kiran Shetty, Hannah Jayne Smith, Imelda McGonnell, Abir Mukherjee Royal Veterinary College, London, UK Follistatin-like 3 (FSTL3), structurally and functionally related to follistatin (FST), is an endogenous inhibitor of TGF[beta] ligands such as activin and myostatin. TGF[beta] ligands are critical not only in adult organ homeostasis but also in development. Moreover, it is becoming increasingly clear that TGF[beta] signalling also plays crucial roles in skeletal development. Activin deletion leads to skeletal changes as well as delayed bone development and deletion of FST is lethal, resulting from several developmental defects including dental, costal and hard palate abnormalities. FSTL3 gene deleted mice (FSTL3 KO) mice survive to adulthood and have been shown to exhibit changes in glucose and lipid metabolism, however the role of FSTL3 in skeletal development and morphology has not previously been investigated. To help define the roles of FSTL3, activin and related TGF[beta] ligand signalling in skeletal development, we investigated the developmental program in FSTL3 KO mice. Homozygous FSTL3 KO (back-crossed to C57/Bl6) and C57/Bl6 (WT) mice were euthanized and embryos harvested at E12.5-E18.5. Embryo and placental weights and crown-rump length were measured. Whole mount skeletal preparations were stained using Alcian Blue and Alizarin Red and adult craniums were studied using microCT. We found that FSTL3 KO animals are larger than their WT counterparts at the earlier developmental timepoints, with the difference normalising at older stages. A marked difference was observed in the bone mineralization program with accelerated calcification of cartilage in FSTL3 KO embryos. Differences were noted in both endochondral and intramembranous bone formation. Cranial and mandibular morphology was also altered showing increased ossification in FSTL3 KO embryos. The difference in cranial morphology persists in the adult FSTL3 KO mice. We are currently monitoring markers of bone development in order to understand the molecular and developmental pathways underlying the observed changes. We are also undertaking micro-CT experiments to measure differences in bone density between WT and FSTL3 KO mice and for a more detailed investigation of embryonic skeletal tissue. Our findings, therefore, clearly demonstrate that FSTL3 gene deletion leads to rapid skeletal tissue development with accelerated bone mineralization and affects cranium development, thus underscoring the importance of regulated activin signalling during development.[br][br]Nothing to Disclose: WM, KS, HJS, IM, AM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2063 319 2515 MON-642 PO17-02 Monday 2184 2012


2180 ENDO12L_MON-643 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) A Novel Role of Follistatin-Like 3 (FSTL3) in Cellular Proliferation Memoona Rehman, Abir Mukherjee Royal Veterinary College, London, UK Follistatin-like 3 (FSTL3) is an endogenous inhibitor that binds and inhibits TGFb family ligands such as activin, myostatin and GDF11. Activin plays crucial roles in development, cellular proliferation and apoptosis while myostatin is a known blocker of muscle growth. To identify physiological roles of FSTL3, FSTL3 gene deletion mice (FSTL3 KO) were generated which displayed altered glucose and lipid metabolism phenotypes. Importantly, the increased pancreatic [beta]-cell proliferation and reduced white adipocyte number observed in FSTL3 KO mice suggest a link between FSTL3 and cellular proliferation and differentiation. To address the role of FSTL3 in cellular biology, an aspect which has not previously been studied, we generated mouse embryonic fibroblasts (MEFs) from FSTL3 KO and FSTL3 conditional/[ldquo]floxed[rdquo] (wildtype, WT) mouse embryos. Using these cells we investigated whether cellular proliferation and signalling are altered as a result of FSTL3 deletion. We found that FSTL3 KO MEFs show significantly increased proliferation in serum fed conditions compared to control WT cells. This difference persists in serum-free culture conditions. To elucidate the cellular signalling pathways that are altered by FSTL3 gene deletion which might affect cellular proliferation, we investigated SMAD and ERK activation profiles in the presence or absence of activin using the MEF cells. In WT MEFs, we found SMAD2 phosphorylation to be significantly enhanced in the presence of exogenous activin, an effect which is reversed following treatment with FSTL3. In the absence of endogenous FSTL3 (KO MEFs), activin-induced SMAD2 phosphorylation is also suppressed after treatment with FSTL3 but the levels of phospho-SMAD2 remain higher compared to WT MEFs, possibly a reflection of reduced inhibition of activin in the absence of endogenous FSTL3. Levels of phosphorylated p42/44 ERK proteins, which play important roles in the regulation of cell growth, were also higher in KO MEFs compared to WT, more so after 1 hour of activin treatment. Taken together, our data demonstrate a novel role for FSTL3 in the regulation of cellular proliferation and suggest that SMAD and ERK signalling pathways might contribute to this function.[br][br]Nothing to Disclose: MR, AM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2243 319 2516 MON-643 PO17-02 Monday 2185 2012


2181 ENDO12L_MON-644 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) Regulation of Hepcidin Expression by BMP Signaling in Macrophages Yin Xia, Xinggang Wu The Chinese University of Hong Kong, Hong Kong, China Hepcidin, a small peptide hormone, negatively regulates iron in circulation by controlling iron absorption from dietary sources and iron release from macrophages. Hepcidin is synthesized mainly in the liver, where hepcidin is regulated by iron loading, inflammation and hypoxia. Recently, we have demonstrated that bone morphogenetic protein (BMP)-hemojuvelin (HJV)-SMAD signaling is central for hepcidin regulation in hepatocytes. Hepcidin is also expressed by macrophages. Studies have shown that hepcidin expression by macrophages increases following bacterial infection, and that hepcidin decreases iron release from macrophages in an autocrine and/or paracrine manner. Although previous studies have shown that lipopolysaccharide (LPS) can induce hepcidin expression in macrophages, whether hepcidin is also regulated by BMPs in macrophages is still unknown. Therefore, we examined the effects of BMP signaling on hepcidin expression in RAW 264.7 and J774 macrophage cell lines, and in primary peritoneal macrophages. We found that BMP4 or BMP6 alone did not have any effect on hepcidin expression in macrophages although they stimulated Smad1/5/8 phosphorylation and Id1 expression. In the presence of LPS, however, BMP4 was able to stimulate hepcidin expression in macrophages, and this stimulation was abolished by the NF-[kappa]B inhibitor Ro1069920. These results suggest that hepcidin expression is regulated differently in macrophages than in hepatocytes, and that BMPs regulate hepcidin expression in macrophages in a LPS-NF-[kappa]B dependent manner.[br][br]Nothing to Disclose: YX, XW 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1331 319 2517 MON-644 PO17-02 Monday 2186 2012


2182 ENDO12L_MON-645 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) Species-Specific Differences in the Expression and Activity of BMP15 Sara L Al-Musawi, Kelly L Walton, Craig A Harrison Prince Henry[apos]s Institute of Medical Research, Melbourne, Australia The mechanism of ovarian folliculogenesis in mammals involves a complex exchange of endocrine signals between the pituitary gland and the ovary and the actions of autocrine/paracrine factors secreted by the oocyte and its surrounding somatic cells. Oocyte-derived BMP15 regulates folliculogenesis, via actions on granulosa cell growth and differentiation. Studies in sheep have also shown that BMP15 is a primary determinant of ovulation rate within species. What is less clear is whether BMP15, given its high evolutionary conservation, could contribute to the marked differences in ovulation rate between species?[br]Similar to other members of the TGF-[beta] superfamily, BMP15 is synthesised as a precursor molecule with the N-terminal prodomain mediating the folding and dimerisation of the C-terminal mature domain. Dimeric precursors are cleaved by proprotein convertases and BMP15 is secreted from the oocyte non-covalently associated with its prodomain. In this study, we show that specific prodomain residues mediate the high level expression of human (h) BMP15, relative to mouse (m) BMP15. Leu[sup]44[/sup]-Glu[sup]50[/sup] of the hBMP15 prodomain lie within the alpha 1 helix, a highly conserved feature amongst TGF-[beta] ligands. Using site-directed mutagenesis to introduce non-conserved mBMP15 residues into this region, we show that Glu[sup]46[/sup] and Glu[sup]47[/sup] are essential for the elevated expression of hBMP15.[br]Following purification, hBMP15 is more potent than mBMP15 at stimulating a luciferase response in COV-434 granulosa cells. BMP15 activity is dependent upon its ability to assemble a signalling complex of type I (ALK6) and type II (BMPRII) receptors. Within the putative type I receptor binding site of BMP15, significant species differences are observed (Arg[sup]329[/sup]/Asp[sup]330[/sup] in human; Pro[sup]329[/sup]/Tyr[sup]330[/sup] in mouse). Substituting Pro[sup]329[/sup]/Tyr[sup]330[/sup] of mBMP15 into hBMP15 reduced bioactivity 5-fold, identifying Arg[sup]329[/sup]/Asp[sup]330[/sup] as mediators of high affinity interactions with ALK6.[br]Collectively, we have identified specific residues in the pro- and mature domains of hBMP15 that enhance growth factor expression and activity, relative to mBMP15. These adaptions may contribute to the variation observed in ovulation rate and fecundity between mammals.[br][br]Sources of Research Support: National Health and Medical Research Council (NHMRC) Project Grant 1006488.[br][br]Nothing to Disclose: SLA-M, KLW, CAH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1372 319 2518 MON-645 PO17-02 Monday 2187 2012


2183 ENDO12L_MON-646 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) Endogenous BMP-7 Signaling Axis in Thyroid Tissues Randa M Abdulrahman, Herman Verloop, Hendrieke C Hoftijzer, Olaf M Dekkers, Guido JC Hovens, Johannes Morreau, Johannes WA Smit Leiden University Medical Center, Leiden, Netherlands; Leiden University Medical Center, Leiden, Netherlands; Leiden University Medical Center, Leiden, Netherlands [bold]Aim[/bold]: Bone morphogenic proteins exhibit multifunctional activities in endocrine tissues (1-3). Therefore, we set out to explore the expression of the BMP-7 intracellular signaling pathway (BMP-7(B7), SMAD-4(S4) and phospho-Smad1 (pS1)) in human thyroid tissues.[br][bold]Materials and Methods[/bold]: We used a tissue array containing 107 benign thyroid tissues (normal thyroid (NT), multinodular Goiter (MNG), and follicular adenoma (FA)) and 83 non medullary thyroid carcinomas (papillary thyroid carcinoma (PTC), follicular thyroid carcinoma (FTC), follicular variant of PTC (FvPTC) and PTC mix FTC). Immunostaining was done for B7, S4 and p-S1. Percentage and intensity of the staining were analyzed and scored semi quantitatively by 3 observers. Differential expression of these 3 proteins was analyzed by a multilevel linear regression estimating their differences among the studied tissues. Bonferroni correction was applied to adjust for multiple testing. Subsequently the same model was applied again for the 3 proteins among the follicular lesions (FTC and FvPTC vs FA).[br][bold]Results[/bold]: The components of the BMP-7 axis were expressed in all thyroid histological variants. In thyroid carcinomas the expression of cytoplasmic (c) - B7, c-S4, c-pS1 was increased (p[lt] 0.001; p[lt]0.01 and p[lt]0.001 vs NT respectively) and the nuclear (n) [ndash]pS1 expression was decreased (p[lt]0.001). In Graves the expression of both c- and n- B7 was decreased (p[lt]0.001 vs NT for both) and the expression of c-S4 and c-pS1 was increased. In FA the expression of c-B7, c-S4 and c-pS1 were higher than in normal tissue. In the MNG only the c-S4 expression was increased. Among the follicular lesions the c-S4 expression was differentially lower in the FTC and FvPTC when compared with FA (p[lt]0.05 and p[lt]0.01 respectively). In FvPTC both n-S4 and n-pS1 were significantly lower in comparison to FA (p[lt]0.001 for both).[br][bold]Conclusion:[/bold] The differential expression of proteins involved in the BMP-7 axis in benign and malignant thyroid tissues points to a role of the BMP-7 axis in thyroid cell differentiation and dedifferentiation. These findings may have relevance both for the pathogenesis of thyroid neoplasms and for the differential diagnosis of thyroid lesions.[br][br](1) Suzuki J et al.,Endocrinology 2004; 145:639. (2) Otsuka F et al., J Biol Chem 2001; 276:32889. (3) Takeda M et al.,Biochem Biophys Res Commun 2003; 306:812.[br][br]Nothing to Disclose: RMA, HV, HCH, OMD, GJCH, JM, JWAS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1489 319 2519 MON-646 PO17-02 Monday 2188 2012


2184 ENDO12L_MON-647 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) Understanding the Mechanism and Partnership of FoxL2 and Smad3 Actions in Pituitary Gonadotropes Alissa N Blackler, Wylie W Vale, Louise M Bilezikjian The Salk Institute for Biological Studies, La Jolla, CA FoxL2 has been shown to play a critical role in gonadotropes by promoting cell-type specific expression of targets including Fshb and Fst. Foxl2 (P-frk) was first identified in the embryonic mouse pituitary and is expressed in differentiated gonadotropes and thyrotropes. In humans and mice, the restricted expression and roles of FOXL2 in the ovary and developing eyelids are well defined. Mutations of FOXl2 cause a syndrome known as BPES, a craniofacial disorder with eyelid malformation and premature ovarian failure (POF). Foxl2 mutant mice similarly display eyelid defects and female infertility. Studies of Foxl2 mutant mice and [alpha]T3-1 and L[beta]T2 cells, which express endogenous FoxL2, have begun to elucidate its pituitary role. We previously demonstrated that activin induces Fst transcription in both cell lines via Smad3 in partnership with FoxL2. In L[beta]T2 cells, the cooperative actions of FoxL2 and Smads also target the Fshb promoter. We have shown that basal and activin-inducible Fshb and Fst expression are compromised in the pituitaries of Foxl2 mutant mice. To better understand the basis for the FoxL2 and Smad3 partnership and their target-specific actions, we have generated a series of deletion and point mutants of FoxL2 and comparatively evaluated the effects of these mutations on activin-responsive rFst- and mFshb-luciferase reporters and on complex formation with Smad3 (mFshb-luciferase reporters provided by Dr. Daniel Bernard, McGill University). Deletion studies indicate that residues within the forkhead domain contribute to the Smad3-binding interface. A deletion mutant that retains the entire forkhead domain retains Smad3 binding, albeit weakly. Interestingly, a somatic mutation found in adult type ovarian granulosa cell tumors, hFOXL2(C134W), localizes to this region. When transiently co-transfected into 293T cells, which are devoid of endogenous FoxL2, hFOXL2(C134W) displayed up to 10-fold or 25-fold higher constitutive activity than wild type on Fst- or Fshb-luciferase reporters, respectively, and caused a further induction of both reporters in the presence of activin A. This mutation similarly affected both reporters in L[beta]T2 cells. The activity of the equivalent mFoxL2(C130W) mutant was similar. This mutation, however, did not have an appreciable effect on Smad3 binding. These studies suggest that differential mechanisms of FoxL2 and Smad3 partnership are involved in target gene activation such as Fst and Fshb.[br][br]Sources of Research Support: NIH grant HD46941.[br][br]Disclosures: WWV: Wylie Vale was a co-founder, consultant, equity holder, membero of the Board of Directors and Scientific Advisory Board., Acceleron Pharma, Neurocrine Biosciences. Nothing to Disclose: ANB, LMB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2350 319 2520 MON-647 PO17-02 Monday 2189 2012


2185 ENDO12L_MON-648 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) The Effects of Ghrelin in Tumor-Induced Cachexia in Mice Are Partially Independent of the GHSR1a Receptor Andres Splenser, Bobby Guillory, Meenal Mendiratta, Tripti Halder, Jose Manuel Garica Baylor College of Medicine, Houston, TX; Michael E DeBakey VAMC, Houston, TX; Texas Children[apos]s Hospital, Houston, TX; Baylor College of Medicine, Houston, TX INTRODUCTION: Ghrelin has been shown to stimulate appetite, increase growth hormone secretion, and decrease inflammatory markers in different settings. Although the only known receptor for ghrelin is the GHSR1a, recent data suggest that not all the effects of ghrelin are mediated through this receptor.[br]OBJECTIVE: To determine if ghrelin[apos]s effects in the setting of tumor-induced cachexia are mediated through the GHSR1a.[br]METHODS: Cachexia was induced by inoculating the Lewis lung carcinoma cell line (LLC) in GHSR-WT and GHSR-KO C57BL/6J male adult mice. Heat-killed cells (HK)-inoculated mice were used as controls. After tumor was palpable, tumor-inoculated mice received either ghrelin (T+G) or vehicle (T+V) intraperitoneally. HK mice also were given vehicle injections. Fat mass (FM), lean body mass (LBM), tumor and liver composition were measured by nuclear magnetic resonance (NMR). A grip meter was used to assess forelimb strength. Animals were sacrificed and tissues and tumors harvested 21 days after injections were started.[br]RESULTS: Tumor inoculation induced a similar degree of fat loss, muscle atrophy, and weakness when KO and WT T+V animals where compared. Ghrelin treatment partially prevented the muscle mass loss and weakness induced by LLC tumor in GHSR-WT and KO animals. Fat mass (NMR and fat pad weights) was significantly decreased by tumor inoculation and this was partially prevented by ghrelin administration in GHSR-WT but not in KO animals. Food intake was no different between groups in either genotype. Finally, ghrelin treatment did not affect tumor size or progression in either genotype.[br]CONCLUSIONS: Treatment with ghrelin partially prevents tumor-induced muscle wasting and weakness in GHSR-WT and GHSR-KO, suggesting that these effects are not mediated through the GHSR1a receptor. On the other hand, ghrelin prevented fat atrophy in this setting only in WT animals, suggesting that these effects are mediated through the GHSR1a. Endogenous GHSR1a signaling does not seem to play an important role in preventing tumor-induced cachexia. In spite of ghrelin[apos]s anabolic properties, tumor growth was unaffected in this model.[br][br]Sources of Research Support: Work funded by the Dept. of Veterans Affairs (BX000507) and by the NIA (AG040583).[br][br]Disclosures: JMG: Investigator, Helsinn Therapeutics, Aeterna Zentaris. Nothing to Disclose: AS, BG, MM, TH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 693 319 2521 MON-648 PO17-02 Monday 2190 2012


2186 ENDO12L_MON-649 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) Glucose Dependence of GH-Induced GHR-JAK2-IGF-1R Protein Complex Formation in Pancreatic Islet Beta-Cells Zhe Wei, Fanxin Ma, James Balducci, Yongchang Chang, Yao Huang St Joseph[apos]s Hospital and Medical Center, Phoenix, AZ; St Joseph[apos]s Hospital and Medical Center, Phoenix, AZ It is known that glucose and several hormones/growth factors, such as growth hormone (GH) and insulin-like growth factor-1 (IGF-1), play important roles in the regulation of pancreatic islet beta-cell mass. However, GH and IGF-1 signaling pathways and their potential crosstalk, especially their relationships to glucose, remain poorly understood. We have recently reported that in multiple rodent pancreatic beta-cell lines, GH specifically promotes the formation of a protein complex containing GH receptor (GHR), Janus kinase 2 (JAK2, a non-receptor kinase coupled to GH/GHR signaling), and IGF-1 receptor (IGF-1R), which may facilitate the crosstalk between GH/GHR and IGF-1/IGF-1R signaling systems (1). Our preliminary work also showed that glucose contributes to the modulation of GH and IGF-1 signaling synergy (2). In this study, we assessed the glucose dependency of GH-induced GHR-JAK2-IGF-1R complex formation, if any, and further explored the potential molecular mechanism. In rat beta-cells (INS-1), using co-immunoprecipitation assay, we demonstrated that acute GH stimulation resulted in the GHR-JAK2-IGF-1R protein complex formation only when the cells were chronically cultured in glucose but not under the condition of their short exposure (15 min) to glucose. This seemed to correlate with high molecular mass of GHR and IGF-1R forms detected on SDS-PAGE. Given that both GHR and IGF-1R are glycoproteins in the cell, we further examined the effects of glucose deprivation and refeeding (dosage and time-course) on the dynamic changes of molecular mass of GHR and IGF-1R. Our data suggested that glucose refeeding was required for the reappearance of high molecular mass of both proteins. Finally, our deglycosylation assay results indicated that the molecular mass changes were indeed due to the glycolytic processes. Taken together, we conclude that the GH-induced GHR-JAK2-IGF-1R protein complex formation requires the presence of glucose and glycosylation (mature forms) of both receptor partners in beta-cells. The biological consequences of such a GH-induced and glucose dependent GHR-JAK2-IGF-1R protein complex, which may have important implications in diabetes, are currently under investigation in our laboratory.[br][br](1) Ma F, et al., Mol Endocrinol 2011; 25: 2119-33. (2) Wei Z, et al., Abstract Book, The Endocrine Society 93rd Annual Meeting, 2011, Boston, MA, USA.[br][br]Sources of Research Support: American Heart Association (AHA) Beginning Grant-in-Aid Award, Science Foundation Arizona (SFAz) Competitive Advantage Award, and St. Joseph[apos]s Foundation (SJF) Startup Fund (to Y.H.).[br][br]Nothing to Disclose: ZW, FM, JB, YC, YH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 117 319 2522 MON-649 PO17-02 Monday 2191 2012


2187 ENDO12L_MON-650 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) IGFBP-2 Stimulates Sustained IGF-I Signaling and Biological Actions Via Inhibition of RPTP[beta] Dephosphorylation of the IGF-I Receptor Xinchun Shen, Gang Xi, Christine Wai, Clifford J Rosen, David R Clemmons The University of North Carolina at Chapel Hill, Chapel HIll, NC; Maine Medical Center Research Institute, Scarborough, NC Activation of the insulin-like growth factor I receptor (IGF-IR) kinase mediates IGF-I-linked intracellular signaling pathways. IGF-IR-mediated functions depend on its phosphorylation status, therefore, IGF-IR dephosphorylation is an important regulatory variable. Previous studies have shown that IGF-I stimulated IGF-IR phosphorylation peaks at 10 min and decreases thereafter. However, no phosphatase has been definitively shown to be responsible for IGF-IR dephosphorylation. RPTP[beta] is a receptor tyrosine phosphatase that binds to IGFBP-2 and mediates its ability to inhibit PTEN dephosphorylation and enzymatic activity thereby and enhancing IGF-I-stimulated AKT activation. However, IGFBP-2 also enhances IGF-I-stimulated MAP kinase (MAPK) and cell proliferation. Therefore we investigated whether there are RPTP[beta] targets other than PTEN. Using RPTP[beta] knockdown in vascular smooth muscle cells, we demonstrated that IGF-I-stimulated IGF-IR phosphorylation was sustained for 60 min after IGF-I treatment (2.6 [plusmn] 0.6-fold increase compared to control). Prolonged IGF-IR tyrosine phosphorylation was associated with sustained MAP kinase and AKT activation in RPTP[beta] knockdown cells. Importantly, no difference of IGF-I-stimulated PTEN tyrosine phosphorylation was detected 60 min after IGF-I treatment when the response of the knockdown cells was compared to control, suggesting that changes in PTEN did not mediate sustained MAPK activation. Purified RPTP[beta] was shown to directly dephosphorylate IGF-IR in vitro. Since we have shown that IGFBP-2 binds to RPTP[beta], leading to its oligomerization and inactivation, we manipulated RPTP[beta] activity by knockdown of IGFBP-2. Loss of IGFBP-2 led to a constitutive increase in RPTP[beta] activity which was associated with impaired IGF-IR phosphorylation following both short-term (10 min) and long-term (60 min) IGF-I treatment. Similar observations were detected in MAP kinase and AKT activation. Importantly, the 60 minute results confirmed that changes in the phosphorylation status PTEN did not mediate changes in RPTP[beta]-linked sustained increases in downstream signaling because IGF-I-stimulated PTEN tyrosine phosphorylation was not detected at this time point. These findings demonstrate that IGFBP-2 inhibition of RPTP[beta] phosphatase activity leads to changes in IGF-IR as well as PTEN. The results suggest that this accounts for the ability of IGFBP-2 to enhance and maintain IGF-I stimulated MAPK activation which leads to enhanced cell proliferation.[br][br]Nothing to Disclose: XS, GX, CW, CJR, DRC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1537 319 2523 MON-650 PO17-02 Monday 2192 2012


2188 ENDO12L_MON-651 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) The Role of Insulin-Like Growth Factor Binding Protein-3 and Poly(ADP-Ribose) Polymerase (PARP) Activation in High Glucose-Induced Apoptosis of Human Proximal Tubular Epithelial Cells Eun-Gyong Yoo, Woo Jung Lee, Hyun Wook Chae, Duk Hee Kim, Ho-Seong Kim CHA University, Sungnam, Korea; Yonsei University, Seoul, Korea Purpose: Insulin-like growth factor binding protein-3(IGFBP-3), the major circulating carrier protein for IGFs, also acts as a potent antiproliferative agent, by blocking cell cycle and inducing apoptosis in various cell types. Recently, we reported that increased IGFBP-3 expression by high glucose mediates high glucose induced apoptosis in LLC-PK1 cells and that increased ROS from high glucose enhances IGFBP-3 expression(1). This study was performed to investigate the role of IGFBP-3 on high glucose induced apoptosis in human proximal tubular epithelial cells (PTEC) and the relationship between poly(ADP-ribose) polymerase (PARP) activation and IGFBP-3 expression.[br]Methods: IGFBP-3 expression and apoptosis in human PTEC line were measured after exposure to either low glucose (5.5mM) or high glucose (30mM, 45mM) media. Apoptosis was quantified by Annexin V flow cytometry. To investigate the specific role of IGFBP-3 on PTEC apoptosis, the effect of small interfering RNA to IGFBP-3 (siRNA:IGFBP-3) and IGFBP-3 overexpression were studied. The relationship between PARP activation and IGFBP-3 expression was also evaluated to investigate the mechanism of high glucose mediated apoptosis in human PTECs.[br]Results: IGFBP-3 protein and mRNA expression was higher in high glucose media compared to low glucose media. Exposure to high glucose media increased PTEC apoptosis, and high glucose-induced PTEC apoptosis was prevented by pre-incubation with siRNA:IGFBP-3. IGFBP-3 overexpression by transfection of IGFBP-3 cDNA induced PTEC apoptosis. High glucose media or IGFBP-3 overexpression induced PARP activation, but the siRNA:IGFBP-3 prevented PARP activation. PARP activation by cisplatin increased IGFBP-3 expression, while 3-aminobenzamide, a PARP inhibitor decreased IGFBP-3 expression.[br]Conclusion: Our results suggest that increased IGFBP-3 expression may mediate apoptosis in human PTEC by high glucose media. Increased IGFBP-3 expression by high glucose is related with activation of PARP.[br][br]Yoo EG et al., Endocrinology 2011;152:3135.[br][br]Nothing to Disclose: E-GY, WJL, HWC, DHK, H-SK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2072 319 2524 MON-651 PO17-02 Monday 2193 2012


2189 ENDO12L_MON-652 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) Distinct Signalling Pathways Involved in the Norepinephrine-Induced Nuclear Accumulation of TORC1 and TORC2 Proteins in Rat Pinealocytes James R McTague, Meghan P Ferguson, Richard A Kanyo, Constance L Chik, Anthony K Ho University of Alberta, Edmonton, Canada; University of Alberta, Edmonton, Canada The transducers of regulated cAMP-respone element-binding protein (CREB)(TORCs) are a family of co-activators that can enhance CREB-mediated gene transcription. Existing literature suggests that for all three TORC isoforms, dephosphorylation and nuclear accumulation are mediated through the same signalling pathways. In this study, we examined whether the identical signalling pathways are involved in the dephosphorylation and nuclear accumulation of TORC1 and TORC2 in the rat pineal gland. Norepinephrine (NE) treatment of cultured pinealocytes leads to the dephosphorylation and nuclear accumulation of TORC1 and TORC2. Through the use of immunoblot analysis and nuclear/cytosolic fractionation we examined that NE activated distinct signalling mechanisms to mediate dephosphorylation and nuclear accumulation of TORC1 and TORC2. Cotreatment with prazosin ([alpha][sub]1[/sub]-adrenergic antagonist) or propanolol ([beta]-adrenergic antagonist) was used to determine which adrenergic pathway was being utilized to mediate the two effects on the specific TORC isoform. NE-mediated nuclear accumulation of TORC1 was prevented with prazosin, while propanolol was capable of preventing the NE-mediated nuclear accumulation of TORC2. NE-mediated nuclear entry of TORC1 was mimicked by elevation of intracellular calcium using a depolarizing concentration of potassium, while dibutyryl cAMP led to nuclear entry of TORC2. NE-mediated dephosphorylation of TORC1 was prevented with cyclosporin A, a protein phosphatase 2B (calcineurin) inhibitor, while okadaic acid, a protein phosphatase 2A inhibitor could prevent the NE-mediated dephosphorylation of TORC2. These results suggest that the NE-mediated dephosphorylation and nuclear accumulation of TORC1 are caused by activation of the [alpha]-adrenergic pathway, with the involvement of elevated intracellular calcium and activation of calcineurin. By contrast, dephosphorylation and nuclear accumulation of TORC2 are caused by activation of the [beta]-adrenergic pathway, which leads to an elevation in cAMP and activation of protein phosphatase 2A. Together, these results highlight the distinct regulatory mechanisms utilized by NE to mediate dephosphorylation and nuclear accumulation of TORC1 and TORC2 in rat pinealocytes.[br][br]Sources of Research Support: Canadian Institutes of Health Research.[br][br]Nothing to Disclose: JRM, MPF, RAK, CLC, AKH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 893 319 2525 MON-652 PO17-02 Monday 2194 2012


2190 ENDO12L_MON-653 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) Sphingosine Kinase 1 Isoform-Specific Interactions Revealed through Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) Eileen M McGowan, Marc R Wilkins, Dominik G Kaczorowski, Pu Xia, Rosetta Martiniello-Wilks, Angelina Lay, Daniel Yagoub University of Technology Sydney, Sydney, Australia; The University of Sydney, Sydney, Australia; University of New South Wales, Sydney, Australia Sphingosine kinase 1 (SK1) is a signaling enzyme that catalyses the formation of sphingosine-1-phosphate. Hormone activation of SK1 is causally associated with breast cancer progression and resistance to therapy. Two major transcriptional isoforms (43 and 51 kDa) of SK1 have been identified, however the 51 kDa variant is predominant in breast cancer cells. No studies have investigated the protein-protein interactions of the 51 kDa isoform, and whether the two SK1 isoforms differ significantly in their interactions. Seeking an understanding of the regulation and role of SK1, we employed a triple-labeling SILAC-based (stable isotope labeling by amino acids in cell culture) approach to identify SK1-interacting proteins common and unique to both isoforms. Of approximately 850 quantified proteins in SK1 immunoprecipitates, a high-confidence list of 30 proteins that interacted with each SK1 isoform was generated via meta-analysis of multiple experimental replicates. Many of the novel identified SK1 interaction partners, including proteins such as supervillin, drebrin, and the MARCKS-related protein supported and highlighted the previously-implicated role of SK1 in breast cancer cell migration, adhesion, and cytoskeletal remodeling. Of these interactions, several were found to be exclusive to the 43 kDa isoform of SK1, including the PP2A phosphatase, a previously identified SK1-interacting protein. Other proteins such as dipeptidyl peptidase 2, allograft inflammatory factor 1-like protein and the latent-transforming growth factor beta-binding protein were found to associate exclusively with the 51 kDa isoform of SK1. In this report, we have identified common and isoform-specific SK1-interacting partners that provide insight into the molecular mechanisms that drive SK1-mediated oncogenicity.[br][br]Nothing to Disclose: EMM, MRW, DGK, PX, RM-W, AL, DY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1057 319 2526 MON-653 PO17-02 Monday 2195 2012


2191 ENDO12L_MON-654 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) Proteomic Exploration of Potential Bioactivities of Proteolytic Peptides of Protein Hormones Kenneth L Campbell, Nurit Haspel, Umaben S Vadher, Jeremy Steinbruck, Noelle Palmstrom, Sherry Solchenberger University of Massachusetts Boston, Boston, MA; University of Massachusetts Boston, Boston, MA Prolactin is known to contain a biologically active peptide vasoinhibin that is released during digestion by target cells. Using proteomic tools we are exploring if many protein hormones thought to act solely via classical transduction mechanisms also generate peptides that have separate actions, e.g., hormonally targeting secondary cells or extending actions of parent hormones by acting as non-classical transducers. We wrote a program, Protein Cyberase, which uses the known lytic sequences for several enzymes, including endosomal/lysosomal cathepsins B, D, G, H, K, and S, and predicts the proteolytic products when these are used singly or in sequence. Validation against other programs was successful for trypsin, CNBR, and, chymotrypsin (sensitivity, specificity, [amp] efficiency = 100% except 99% specificity for chymotrypsin). Sequences of 4-6 cathepsins with pH optima of 7.5 to 3.5, imitating the enzyme exposure within endosomal-lysosomal compartments, have been applied to [gt]20 protein hormones and biomarkers of nidation. The hormones examined produce 0-[gt]20 cathepsin-resistant peptides of [gt]6 residues (most peptides are 6-15 residues but MUC1 produced [gt]20 peptides of [gt]20-24 residues). Some peptide locations in glycosylated proteins, e.g., hCG[beta], suggest part of the predicted peptides may remain uncleaved if proteolysis precedes deglycosylation during cellular digestion. FSH[beta] peptides have been modeled for 3-D structure. These motifs were then matched to proteins with x-ray structures in the PDB. Many of these motifs lie at the surfaces of the matched proteins and may be active in protein-protein interactions; this is being explored using protein network-neighborhood software. Sequential motifs matching the cathepsin-resistant peptides have been explored using BLASTp (local alignments, PAM 45, 0 gap). While short peptides elevate background noise in PDB searches and limit E value utility for picking best matches, we have usually found an array of proteins sharing the linear motifs. Matches include non-parent protein hormones, receptors for other hormones, cytoplasmic enzymes, transduction proteins, and transcription factors. The lists of sequential and 3-D motif matches and their protein network partners provide candidates for biochemical and genetic work on the role and scope of bioactivities that may be innate to protein hormone proteolytic peptides.[br][br]Sources of Research Support: In part by a Healey Endowment Grant (NH and KLC) and an Office of International and Transnational Affairs travel grant (KLC) from University of Massachusetts Boston.[br][br]Nothing to Disclose: KLC, NH, USV, JS, NP, SS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2324 319 2527 MON-654 PO17-02 Monday 2196 2012


2192 ENDO12L_MON-655 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) Reduction of Phosphoinositide 3-OH Kinase p110 Beta Is Associated with Increased Myoblast Differentiation Ronald W Matheny, Christine M Lynch United Stated Army Research Institute of Environmental Medicine, Natick, MA Phosphoinositide-3-OH kinase (PI3K) has been shown to regulate skeletal muscle cell differentiation; however, the roles of individual PI3K p110 catalytic subunits in myogenesis are less well-understood. To determine the potential roles for PI3K p110 alpha and p110 beta catalytic subunits, we first defined the mRNA transcript expression patterns of p110 alpha and p110 beta during muscle differentiation in C2C12 cells. We found that levels of p110 alpha were elevated [sim]50% during differentiation, with a maximal expression two days after induction of differentiation (P [lt] 0.05). Conversely, levels of p110 beta declined [sim]50% during differentiation, with a minimal expression two days following onset of differentiation (P [lt] 0.05). The abundance of both p110 alpha and p110 beta returned to levels seen at induction of differentiation (0 days) 96 hours after onset of differentiation. Since reduced levels of p110 beta were associated with myogenesis, we next treated myoblasts with siRNA against p110 beta to determine whether reductions in p110 beta would accelerate the myogenic program. We observed that p110 beta-deficient myoblasts exhibited increased levels of myogenic proteins including MyoD ([sim]2.5-fold increase; P [lt] 0.01) and myogenin ([sim] 8-fold increase; P [lt] 0.01). Additionally, under differentiation conditions for 24 or 48 hours, protein levels of MyoD and myogenin were elevated beyond control levels. Together, these data demonstrate that reductions in p110 beta are associated with markers of increased myogenesis and suggest a negative role for p110 beta during myoblast differentiation.[br][br]Sources of Research Support: This work was supported by an appointment to the postgraduate research participation program at the United States Research Institute of Environmental Medicine administered by the Oak Ridge Institute for Science and Education through an interagency agreement between the United States Department of Energy and Unites States Army Medical Research and Materiel Command (C.M.L).[br][br]Nothing to Disclose: RWM, CML 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1567 319 2528 MON-655 PO17-02 Monday 2197 2012


2193 ENDO12L_MON-656 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) Guanylyl Cyclase Receptor-A Signaling Inhibits Agonist-Stimulated MAPKs and Downstream Effectors AP-1 and CREB Satyabha Tripathi, Kailash N Pandey Tulane University Health Sciences Center, New Orleans, LA The cardiac hormone, atrial natriuretic peptide (ANP) and its guanylyl cyclase/natriuretic peptide receptor-A (GC-A/NPRA) exhibit an inhibitory effect on cell growth and proliferation, along with natriuretic, diuretic, and vasodilatory properties. However, the ANP/NPRA signaling pathways mediating the antimitogenic effects are not well understood. The objective of this study was to determine the effect of ANP-NPRA system on mitogen-activated protein kinases (MAPKs) and the downstream proliferative transcription factors involving activating protein-1 (AP-1) and cAMP-response element binding protein (CREB) in agonist-stimulated mouse mesangial cells (MMCs). We found that ANP inhibited vascular endothelial growth factor (VEGF)-stimulated phosphorylation of MAPKs (Erk1, Erk2, JNK, and p38), to a greater extent in NPRA-transfected cells (50-60%) relative to vector-transfected cells (25-30%). The analyses of the phosphorylated transcription factors revealed that ANP inhibited VEGF-stimulated activation of CREB, and the AP-1 subunits (c-jun and c-fos). Gel shift assays demonstrated that ANP inhibited VEGF-stimulated AP-1 and CREB DNA-binding ability by 67 % and 62 %, respectively. The addition of the protein kinase G (PKG) inhibitor, KT-5823, restored the VEGF-stimulated activation of MAPKs, AP-1, and CREB, demonstrating the integral role of cGMP/PKG signaling in NPRA-mediated effects. Our results delineate the underlying mechanisms through which ANP-NPRA system exerts an inhibitory effect on MAPKs and down-stream effector molecules, AP-1 and CREB, critical for cell growth and proliferation.[br][br]Sources of Research Support: National Institutes of Health grant HL57531.[br][br]Nothing to Disclose: ST, KNP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1143 319 2529 MON-656 PO17-02 Monday 2198 2012


2194 ENDO12L_MON-657 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) Decreased Endocytosis of Guanylyl Cyclase/Natriuretic Peptide Receptor-A by RNA Interference: Role of microRNA Naveen K Somanna, Amitabh C Pandey, Kailash N Pandey Tulane University Health Sciences Center, New Orleans, LA Guanylyl cyclase/natriuretic peptide receptor-A (GC-A/NPRA) is the principal locus for the regulatory action of atrial and brain natriuretic peptides (ANP and BNP) and plays a critical role in control of extracellular fluid volume and blood pressure homeostasis. We have used microRNA (miRNA)-mediated small interfering RNA (siRNA) for functional gene-knockdown of GC-A/NPRA in stably transfected human embryonic kidney 293 (HEK-293) cells expressing a high density of recombinant GC-A/NPRA. We artificially expressed three RNA polymerase II-driven miRNAs that specifically targeted the [italic]Npr1[/italic] gene (coding for GC-A/NPRA), but shared no significant sequence homology with any other known mouse genes. Reverse transcription-PCR (RT-PCR) and Northern blot analysis identified two highly efficient [italic]Npr1[/italic] miRNA sequences to knockdown the expression of NPRA. Furthermore, [italic]Npr1[/italic] miRNA in chains or clusters decreased NPRA expression more than 90% compared with control cells. ANP-stimulated intracellular accumulation of cGMP was significantly reduced in [italic]Npr1[/italic] miRNA-expressing cells. Treatment with [italic]Npr1[/italic] miRNA caused a drastic reduction in the internalization of radiolabeled [sup]125[/sup]I-ANP-NPRA ligand-receptor complexes. Only 15%-20% of receptor population was internalized in micro-RNA silenced cells as compared to 70%-80% receptors in control cells. The results demonstrate that RNA interference can silence the expression of [italic]Npr1[/italic]gene[italic],[/italic] as a result, the internalization and trafficking of NPRA is drastically reduced in intact cells.[br][br]Sources of Research Support: National Institutes of Health Grant HL 57531.[br][br]Nothing to Disclose: NKS, ACP, KNP 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1165 319 2530 MON-657 PO17-02 Monday 2199 2012


2195 ENDO12L_MON-658 POSTER SESSION: Signaling in Development [amp] Differentiation (1:30 PM-3:30 PM) Spot Determinations of Urinary 3-Methylhistidine Isotope Enrichment Decay To Identify Age-Related Differences in Protein Degradation James Lynch, William Durham, Mort Jangorbani, Sally Schuette, Edgar Dillon, Christopher Danesi, Katherine Randolph, Shanon Casperson, John Rathmacher, Astrid Horstman, Greg White, Randall Urban, Melinda Moore University of Texas Medical Branch, Galveston, TX; University of Texas Medical Branch, Galveston, TX; BioChemAnalysis Corporation and Center for Stable Isotope Research Inc, Chicago, IL; Metabolic Technologies Inc, Ames, IA Background: 3-methylhistidine (3MH) is released during myofibrillar degradation and quantitatively excreted in the urine, allowing for collection and measurement of 24-hour urinary [3MH] to be used as a proxy for estimation of skeletal muscle proteolysis. 3MH is traditionally measured in 24-hour urine samples following at least three days of abstinence from consumption of meat-containing meals. Here, we sought to determine 1) the relationship between spot measures of 3MH isotope enrichment decay in plasma vs. urine, 2) the effect of meat vs. non-meat containing meals prior to sample collection, and 3) the effect of age on 3MH excretion, in order to determine whether urinary 3MH enrichment decay analysis represents a viable alternative approach to assessing skeletal muscle proteolysis.[br]Methods: Healthy older (n=5, 66[plusmn]4 years, BMI 26[plusmn]1 kg/m2) and younger (n=4, 33[plusmn]1 years, BMI 27[plusmn]1 kg/m2) volunteers were studied on two occasions: following consumption of a meat-containing evening meal and following consumption of a vegetarian evening meal. Meal order was randomly determined and macronutrient proportions of the meat and vegetarian meals were matched. With each evening meal, the subject ingested a dose of isotopically labeled (d3-methyl) 3MH mixed into a small volume of diet soda. Plasma and spot urine samples were collected 13 to 22 hours after dosing. All samples were processed to determine the 3MH tracer/tracee ratios and decay curves vs. time constructed.[br]Results: Isotope ratios were similar plasma or urine measures or following meat vs. meatless meals. The mean slopes of the urinary terminal decay curves were -0.0771 and -0.0575 for the older and younger subjects respectively, with no overlap in values between the groups. The slope of the decay curve was significantly greater (i.e. more negative) in the older subjects (p[lt]0.025), suggesting greater de novo 3MH production due to skeletal muscle proteolysis in the older subjects.[br]Conclusion: The difference in the calculated rate constant for de novo 3MH production between the older and younger subjects suggests that spot urine collection may be sufficiently sensitive to measure differences in the rates of myofibrillar protein breakdown between individuals. Thus, with this approach, it may be possible to non-invasively identify individuals with elevated rates of muscle proteolysis (e.g. cancer and older hospitalized patients) before a significant loss of muscle mass occurs.[br][br]Sources of Research Support: NIH Grant 1R43 AR054993-01.[br][br]Nothing to Disclose: JL, WD, MJ, SS, ED, CD, KR, SC, JR, AH, GW, RU, MM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2102 319 2531 MON-658 PO17-02 Monday 2200 2012


2196 ENDO12L_MON-664 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Simultaneous Stimulation of Human 20 kDa Growth Hormone and 22 kDa Growth Hormone Secretion by Ghrelin Jenny Tong, David D[apos]Alessio, Juliane Ramisch, Harold Davis, Matthias Tschoep, Martin Bidlingmaier University of Cincinnati, Cincinnati, OH; Cincinnati VA Medical Center, Cincinnati, OH; Ludwig-Maximilians Universitat, Munich, Germany 20 kDa human growth hormone (hGH) is produced in the pituitary by alternative splicing of the hGH-N gene. Its growth promoting ability is equal to 22 kDa hGH, the predominant form in circulation, but the effect on glucose metabolism has been debated.[br]Objective: To investigate the effect of ghrelin on 20 kDa and 22 kDa hGH secretion in healthy, non-obese subjects. In addition, we aimed to study the association between endogenous GH concentration and fating plasma glucose and insulin as well as the relationship between dynamic GH secretion and insulin sensitivity.[br]Design and Setting: Synthetic human acyl ghrelin (0.2 or 0.6 nmol/kg/hr) or saline was infused in a randomized order in 14 healthy subjects (6 M/8 F; age 27.7 [plusmn] 6.3 years; BMI 22.0 [plusmn] 2.7 kg/m[sup]2[/sup], mean [plusmn] SEM) on three separate days. Ghrelin was infused for 45 minutes to achieve steady-state levels and continued through a 3-h frequently sampled IV glucose tolerance test (FSIGT). Insulin sensitivity index (S[sub]I[/sub]) was quantified using the minimal model of glucose kinetics.[br]Results: At baseline, the 20 kDa and 22 kDa GH concentrations were 0.35 [plusmn] 0.07 and 2.2 [plusmn] 0.4 ng/ml, respectively, with a ratio of 20 kDa:22 kDa GH of 0.13 [plusmn] 0.02. Females had significantly higher baseline GH levels than males. Ghrelin administration increased serum 20 kDa and 22 kDa GH levels in a parallel and dose dependent fashion, with no significant change in the 20kDa:22kDa GH ratio. Both baseline 20 kDa and 22 kDa GH levels were negatively correlated with fasting plasma glucose, insulin and HOMA-IR. During the FSIGT, GH secretion (AUC[sub]20 kDa GH[/sub] and AUC[sub]22 kDa GH[/sub]) was positively correlated with insulin sensitivity as measured by S[sub]I[/sub].[br]Conclusion: Our study showed that ghrelin dose dependently increased endogenous 20 kDa and 22 kDa GH secretion in healthy subjects, but did not affect the ratio of these two GH isoforms. Both circulating levels and secretion of these two GH isoforms are similarly associated with and regulated by glucose and insulin.[br][br]Sources of Research Support: NIH/NIDDK K23DK80081, R03DK89090.[br][br]Nothing to Disclose: JT, DD, JR, HD, MT, MB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1688 320 2532 MON-664 PO18-02 Monday 2201 2012


2197 ENDO12L_MON-665 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) The Magnitude of Growth Hormone Pulse Amplitude Helps Regulate the Elevated Lipolytic Rate during Prolonged Fasting Naila Goldenberg, Jeff Horowitz, Ashraf Gorgey, Alla Sakharova, Sowmya Surya, Ariel Barkan University of Michigan, Ann Arbor, MI; University of Michigan, Ann Arbor, MI; University of Michigan, Ann Arbor, MI [bold]Background[/bold]: Growth hormone (GH) is an important hormonal regulator of metabolism during fasting. During a prolonged fast, insulin secretion is reduced and GH secretion is increased, and we have previously reported that GH pulsatility played a key role in the elevated lipolytic rate found during a prolonged fast.[br][bold]Study Aims[/bold]: To determine if changes in the magnitude of GH pulse amplitude affect lipolytic rate during prolonged fasting, and to assess the relationship between lipolytic rate and discrete components of GH pulsatility (i.e., maximum pulse amplitude, total pulse area, pulse frequency, and interpulse GH concentration).[br][bold]Study Protocol[/bold]: 15 young, healthy and non-obese subjects (11 men and 4 women) volunteered for this study. We measured lipolytic rate (using stable isotope dilution techniques) after an overnight fast ([bold]12h Fast[/bold]; n=15) and again on four separate occasions after a 52h fast. These 52h fasting trials differed only by the dose of a selective GH Releasing Hormone antagonist (GHRH-A) administered throughout the fast to suppress GH secretion to different levels; [bold]No GHRH-A[/bold] (n=15); [bold]HIGH[/bold] (10 Mcg/kg/h, n=15), [bold]MEDIUM[/bold] (1 Mcg/kg/h, n=8), [bold]LOW[/bold] (0.5 Mcg/kg/h, n=6)[br][bold]Results: [/bold]As expected, the 52h fast markedly increase endogenous GH concentration ([bold]No GHRH-A [/bold]vs [bold]12h Fast[/bold]; P=0.0044). GHRH-A induced dose-dependent reduction in GH pulse amplitude during the 52h fast with more than an 80% suppression during [bold]HIGH[/bold]. After 52h of fasting, we found a strong correlation between lipolysis and GH maximum peak amplitude (R=0.54; P=0.0003), and total pulse area (R=0.49; P=0.0015), but not the GH peak frequency (R=0.04; P=0.8) or interpulse GH concentrations (R=0.2; P=0.1).[br][bold]Summary: [/bold]Lipolytic rate at the end of a 52h fast was directly related to magnitude of the GH pulse amplitude and pulse area. GH pulse frequency and interpulse GH levels did not affect lipolysis during the prolonged fast.[br][br]Sources of Research Support: NIH Grant # R01 DK071955Genentech[apos]s protocol number X3739n; Grant # 805-F06.[br][br]Nothing to Disclose: NG, JH, AG, AS, SS, AB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 876 320 2533 MON-665 PO18-02 Monday 2202 2012


2198 ENDO12L_MON-666 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Distinct Metabolic Surrogates Determine Basal and Rebound GH Secretion after Glucose Ingestion in Men Ali Iranmanesh, Donna Lawson, Johannes D Veldhuis Salem Veterans Affairs Medical Center, Salem, VA; Mayo Clinic, Rochester, MN [bold]Context[/bold]. Growth hormone (GH) secretion declines rapidly after glucose ingestion, and then recovers to higher than baseline levels (rebound GH release).[br][bold]Hypothesis[/bold]. Metabolic factors selectively regulate the biphasic GH response to glucose.[br][bold]Design[/bold]. Prospectively randomized crossover study of GH secretion after glucose vs water ingestion.[br][bold]Setting[/bold]. Clinical Translational Research Center.[br][bold]Participants[/bold]. Sixty-nine healthy men ages 19-78 yr with body mass index (BMI) 18-39 kg/m[sup]2[/sup].[br][bold]Outcomes[/bold]. Nadir vs peak GH concentrations and basal vs pulsatile GH secretion.[br][bold]Results[/bold]. Mean nadir GH concentrations were determined positively by SHBG after glucose administration (R[sup]2[/sup]=0.088, P=0.0077). Peak rebound GH concentrations were related positively to adiponectin and negatively to CT-estimated abdominal visceral fat (AVF) (R[sup]2[/sup]=0.182, P=0.00049) after glucose ingestion. Deconvolution analysis showed that SHBG specifically predicted basal (nonpulsatile) GH secretion following glucose exposure (R[sup]2[/sup]=0.153, P=0.00052). In contrast, all three of exercise history and adiponectin (both positively) and AVF (negatively) predicted pulsatile GH escape after glucose suppression (R[sup]2[/sup]=0.206, P=0.00043). Moreover, adiponectin uniquely determined the size (mass), and AVF the mode (duration), of GH secretory bursts after glucose exposure (both P[lt]0.006).[br][bold]Conclusion[/bold]. Glucose ingestion provides a clinical model for investigating complementary metabolic surrogates that determine both basal and pulsatile GH secretion in healthy men.[br][br]Sources of Research Support: In part via the Center for Translational Science Activities (CTSA) Grant Number 1 UL 1 RR024150 from the National Center for Research Resources (Rockville, MD), and AG19695 and DK050456 (Metabolic Studies Core of the Minnesota Obesity Center) from the National Institutes of Health (Bethesda, MD). Supported by the Salem Veterans Affairs Medical Center and Salem Research Institute (Salem, Virginia).[br][br]Nothing to Disclose: AI, DL, JDV 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 168 320 2534 MON-666 PO18-02 Monday 2203 2012


2199 ENDO12L_MON-667 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) The Correlation between Growth Hormone, Hand-Grip Strength and Body Composition in Elderly People Ignacio Antonio Seixas-da-Silva, Ana Beatriz Winter Tavares, Maria Lucia Fleiuss de Farias, Mario Vaisman, Flavia Lucia Conceicao Federal University of Rio de Janeiro, Rio de Janeiro, Brazil Introduction: During the aging process occur hormones modifications that modify the metabolism of the individuals diminishing their functional capacities. The hormones of the anabolic way, p.ex, growth hormone (GH), are the ones that present biggest alterations and the reduction of the production of these hormones is an event that reaches the aged ones, generating modifications that intervene with the activities of daily life. Objective: This study aims to correlate the values of body composition, hand-grip strength and the GH peak secretion. Methods: The sample was composed by 16 individuals, being 12 females and 4 males; the glucagon stimulus test was made with injection, via intramuscular, of 1mg of glucagon with collection of blood after 90, 120, 150 and 180 minutes. For the hand-grip strength test, a JAMAR[reg] dynamometer was used and the final measure, in kilogram-force (kgf), was the mean between three measures carried through with the dominant hand of the individual. The body composition was carried through the Dual-energy X-ray Absorptiometry (DXA), using Lunar Prodigy Advance[reg] densitometry, where the values of percentage of fat (% G), lean mass (kg) and fat mass (kg) had been gotten, the total body mass and stature had been gotten through the Filizola[reg] scale. For descriptive analysis was used mean and standard deviation and the Pearson[apos]s correlation coefficient were used to find the relations between the studied variables. Results: The sample had age of 78,3 [plusmn] 5,66 years-old, total body mass of 63,8 [plusmn] 8,9 kg, stature of 1,54 [plusmn] 0,06 meters and Body Mass Index (BMI) of 27,1 [plusmn] 316 kg/m[sup2]. The mean percentage of fat found was 40,4 [plusmn] 4,98 %, lean mass 37,2 [plusmn] 5,4 kg and fat mass 24,4 [plusmn] 4,98 kg. To the hand-grip strength and to the peak response of GH secretion to the glucagon stimulus, the value found were, respectively, 11,39 [plusmn] 6,38 kgf and 5,3 [plusmn] 2,49 mcg/L. A regular inverse correlation were found between the lean mass and the GH peak (r = -0,546) and between the hand-grip strength and the GH peak (r = -0,576). A regular positive correlation between the lean mass and the hand-grip strength was found (r = 0,553). Conclusion: On the basis of the found results, it[apos]s possible to conclude that for the elderly exists a relation between the secretion of the GH and the body composition in set with the muscular strength, making believe that a reduction of the GH secretion may cause a reduction of the functional capacity.[br][br]Nothing to Disclose: IAS-d-S, ABWT, MLFdF, MV, FLC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2383 320 2535 MON-667 PO18-02 Monday 2204 2012


2200 ENDO12L_MON-668 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Positive Associations between Serum Levels of IGF-I and Subcutaneous Fat Depots in Young Men. The Odense Androgen Study Marianne Andersen, Kim Brixen, Claus Hagen, Jan Frystyk, Torben Leo Nielsen Odense University Hospital, Odense, Denmark; Aarhus University Hospital, Aarhus, Denmark [bold]Introduction[/bold][br]Serum levels of IGF-I are of growing interest due to the associations with diseases closely related to obesity. Despite markedly suppressed GH secretion, total IGF-I levels are often within normal range in obese adults.[br][bold]Aim[/bold][br]To study associations between IGF-I and estimated muscle mass in the Odense Androgen Study population and secondly to investigate associations between serum IGF-I and regional fat depots.[br][bold]Method[/bold][br]The Odense Androgen Study is a population-based, cross-sectional study of 776 randomly selected men aged 20-29 years. Regional lean and fat mass were measured by dual-energy x-ray absorptiometry, whereas regional muscle and fat areas were assessed by magnetic resonance imaging.[br][bold]Results[/bold][br]Age-adjusted IGF-I levels correlated significantly with different estimates of muscle mass (r-values between 0.15 and 0.19; p[lt]0.001). Using multiple linear regression, serum IGF-I correlated positively with subcutaneous adipose tissue on the abdomen (SAT) after controlling for visceral adipose tissue (VAT) in the whole group and in the subgroup of men with normal waist circumference (r-values between 0.13 and 0.15; p[lt]0.03). Accordingly, IGF-I correlated positively with subcutaneous thigh fat area after controlling for intramyocellular lipid (r=0.18; p[lt]0.004).[br][bold]Conclusion[/bold][br]SAT and subcutaneous thigh fat areas were positively associated with IGF-I in regression analyses. Conversely, intramyocellular lipid of the thigh was inversely associated with IGF-I levels. Thus, further insight in the associations between regional fat depots and IGF-I levels may improve our understanding of the GH/IGF-I system.[br][br]Sources of Research Support: Novo Nordisk Foundation, WADA, University of Southern Denmark (SDU).[br][br]Nothing to Disclose: MA, KB, CH, JF, TLN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1102 320 2536 MON-668 PO18-02 Monday 2205 2012


2201 ENDO12L_MON-669 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Effects of Raloxifene and Estrogen on Bioactive IGF-I in Healthy and GH-Deficient Women Vita Birzniece, James Gibney, Ken Ho, Jan Frystyk St Vincent[apos]s Hospital, Garvan Institute of Medical Research, Sydney, Australia; Aarhus University Hospital, Aarhus, Denmark GH action is attenuated by oral estrogens and selective estrogen receptor modulators (SERMs). During GH therapy in hypopituitary women, co-treatment with raloxifene, a SERM, leads to a smaller gain in lean body mass (LBM) than with estrogen (1). During raloxifene co-treatment, circulating IGF-I was similar while IGFBP-3 higher than with estrogen co-treatment. We hypothesized that a higher IGFBP-3 concentration reduces IGF-I bioactivity causing an attenuated anabolic effect observed during raloxifene treatment.[br]We compared bioactive IGF-I to immunoreactive (IR) IGF-I in two previous studies (2, 3). In the first study, 9 healthy postmenopausal and 12 GHD women were randomised to raloxifene 120 mg/d or 17[beta]-estradiol (E2) 4 mg/d for 1 month. In the second study, 17 GHD women were randomised to 1 month treatment with GH alone (0.5 mg/d) and in combination with raloxifene (60 mg/d) or E2 (2 mg/d). We measured bioactive IGF-I by a kinase receptor activation assay, KIRA (4) and IR IGF-I.[br]In study 1, the mean bioactive IGF-I concentration was significantly (p[lt]0.05) lower in GHD than in normal women. In both groups bioactive IGF-I fell with raloxifene and E2 with no significant difference between treatments. In study 2, GH therapy significantly increased mean bioactive IGF-I concentration which fell during raloxifene ([Delta] -23[plusmn]7 %, p[lt]0.01) and E2 co-treatments ([Delta] -26[plusmn]13 %, p=0.06), with the change not significantly different between co-treatments. As previously reported, circulating IGFBP-3 was significantly higher during raloxifene (by 25% in study 1 and by 37% for study 2) compared to E2 treatment. There was a significant correlation (R2 = 0.54, p[lt]0.001) between the two measurements, with bioactive being 3.1[plusmn]0.2 % that of IR IGF-I. There was no significant difference (p=0.2) in the proportion of bioactive IGF-I between raloxifene (2.9[plusmn]0.3 % of IR IGF-I) and E2 (3.4[plusmn]0.3 % of IR IGF-I) treatments.[br]In summary, E2 and raloxifene reduced bioactive and IR IGF-I levels equally while IGFBP-3 levels were higher during raloxifene treatment. The bioactive to IR ratio is unaffected by raloxifene treatment. We conclude that the difference in the effects on LBM between raloxifene and E2 treatments cannot be explained by differences in IGF-I bioactivity. The anabolic attenuating effect of raloxifene is unlikely to involve IGF-I mediation.[br][br](1) Birzniece et al. 2012 J Clin Endocrinol Metab PMID: 22170716, In Press. (2) Birzniece et al. 2010 J Clin Endocrinol Metab 95:2099-106. (3) Gibney et al. 2005 J Clin Endocrinol Metab 90:3897-903. (4) Chen et al. 2003 Am J Physiol Endocrinol Metab 284:E1149-55.[br][br]Sources of Research Support: The NHMRC and by Lilly Australia.[br][br]Nothing to Disclose: VB, JG, KH, JF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 531 320 2537 MON-669 PO18-02 Monday 2206 2012


2202 ENDO12L_MON-670 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Negative Regulation of 11[beta]-HSD1 by GH in Key Metabolic Tissues May Contribute to Metabolic Disease in GH- Deficient Patients Stuart Andrew Morgan, Darlene E Berryman, Edward O List, Gareth G Lavery, John J Kopchick, Paul M Stewart University of Birmingham, Birmingham, UK; Ohio University, Athens, OH Patients with growth hormone deficiency (GHD) have many clinical features in common with Cushing[apos]s syndrome (glucocorticoid excess) [ndash] notably visceral obesity, insulin resistance, osteoporosis and increased vascular mortality. Within key metabolic tissues, 11[beta]-hydroxysteroid dehydrogenase type 1 (11[beta]-HSD1) converts cortisone to the active glucocorticoid, cortisol, and thus amplifies local glucocorticoid action. We hypothesise that 11[beta]-HSD1 expression is negatively regulated by growth hormone (GH), and that GHD patients have elevated 11[beta]-HSD1 levels (leading to increased intracellular cortisol generation) which contributes to the clinical features of this disease. To identify the impact of GH excess/deficiency on 11[beta]-HSD1 in vivo, we measured mRNA expression in key metabolic tissues of giant mice expressing the bovine GH (bGH) gene, dwarf mice with a disrupted GH receptor (GHRKO) gene and mice expressing a gene encoding a GH receptor antagonist (GHA).[br]In male bGH mice, 11[beta]-HSD1 expression was decreased in gastrocnemius muscle (0.43-fold, p[lt]0.05), epididymal adipose tissue (0.38-fold, p[lt]0.05) and subcutaneous adipose tissue (0.57-fold, p[lt]0.05), but not liver. By contrast, male GHRKO mice had increased 11[beta]-HSD1 expression in the liver (2.8-fold, p[lt]0.05), with no differences in expression levels in gastrocnemius muscle, epididymal adipose tissue or subcutaneous adipose tissue. Likewise, male GHA mice had increased 11[beta]-HSD1 expression in liver (2.3-fold, p[lt]0.05), with no difference in expression in gastrocnemius muscle, epididymal adipose tissue or subcutaneous adipose tissue.[br]In summary, we have found GH to negatively regulate 11[beta]-HSD1 expression in a tissue specific manner. As such, providing evidence that increased intracellular cortisol production within key tissues may contribute to metabolic disease in GHD patients.[br][br]Nothing to Disclose: SAM, DEB, EOL, GGL, JJK, PMS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1458 320 2538 MON-670 PO18-02 Monday 2207 2012


2203 ENDO12L_MON-671 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Effects on Insulin Sensitivity and Body Composition of Combination Therapy with Growth Hormone (GH) and Insulin-Like Growth Factor-I (IGF-I) in GH-Deficient Adults with Diabetes Caroline Barner, Maria Petersson, Britt Eden Engstrom, Charlotte Hoybye Sct G[ouml]rans Hospital, Stockholm, Sweden; Karolinska University Hospital, Stockholm, Sweden; Uppsala University Hospital, Uppsala, Sweden [bold]Context:[/bold] Growth hormone (GH) treatment has been avoided in some GH deficient (GHD) adults with diabetes mellitus because of GH[apos]s insulin-antagonistic effects on glucose metabolism.[br][bold]Objective:[/bold] To evaluate the effect on glucose sensitivity and body composition of combination therapy with GH and insulin-like growth factor I (IGF-I) in adults with GHD and diabetes.[br][bold]Design, Patients:[/bold] The pilot study was a 6 months randomised placebo controlled trial in 14 adults with GHD and type 2 diabetes. All received rhGH (0.15 mg/day for 1 month, 0.3 mg/day for 5 months) and were randomised to rhIGF-I or placebo (15 [micro]g/kg/day for 1 month and 30 [micro]g/kg/day for 5 months). Glucose metabolism was evaluated with euglycemic hyperinsulinemic clamp and body composition by computed tomography (CT) of abdominal and thigh fat and bioimpedance.[br][bold]Results:[/bold] IGF-I and IGF-I standard deviation score (SDS) increased in both groups, with the highest increase (median 276 [micro]g/L and 4.8) in the GH and IGF-I group. The patients were obese and at baseline insulin sensitivity (M-value) was low. Positive changes in M-value by +1.4 mg/kg/min, in subcutaneous abdominal fat by -60.5 ml and in fat free mass by +4 percent were seen in the GH and IGF-I group, corresponding values in the GH and placebo treated group were -1.5 mg/kg/min, +23 ml and -0.04 percent respectively (p=0.02, p=0.04 and p=0.03 for delta values between groups). No safety issues occurred.[br][bold]Conclusions:[/bold] Effects of the combined treatment were positive but rather limited and it might be an advantageous treatment option only in a few selected patients.[br][br]Sources of Research Support: Grants from IPSEN Sweden.[br][br]Nothing to Disclose: CB, MP, BEE, CH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1239 320 2539 MON-671 PO18-02 Monday 2208 2012


2204 ENDO12L_MON-672 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Relationship between IGF-1 and Anemia in Older Subjects: Cross Sectional and Longitudinal Data from the InCHIANTI Study Marcello Maggio, Fulvio Lauretani, Chiara Cattabiani, Stefano Parrino, Francesca Re, Simonetta Morganti, Graziano Ceresini, Alessandro Vignali, Silvia Masoni, Stefania Bandinelli, Luigi Ferrucci, Gian Paolo Ceda University Hospital of Parma, Parma, Italy; University Hospital of Parma, Parma, Italy; University Hospital of Parma, Parma, Italy; Azienda Sanitaria of Florence, Florence, Italy; National Institute on Aging, Baltimore, MD [bold]Background.[/bold] The prevalence of anemia is higher in older people, where predicts disability and mortality(1).Among different potential mechanisms involved in the anemia of aging attention has been devoted to the decline of anabolic hormones especially testosterone(T)(1,2).IGF-1 can also contribute to the age-related decline in hemoglobin (Hb) levels(3),but only one study in adult population has tested the link between low IGF-1 and anemia(4).Thus,we addressed this hypothesis in older subjects.[br][bold]Materials and Methods[/bold].The population included 402 men, 536 women 65 yr older of the InCHIANTI Study with data on Hb, IGF-1 levels and T at baseline and Hb after 3 years.Serum IGF-1 were measured by IRMA.The MDC was 0.80 ng/mL with CVs[lt]10%. T was measured by RIA.The MDC for T was 0.03 nmol/L,the intra and interassay CV were[lt]10%.The MDC was 0.1 pg/mL and interassay CV of 4.5%. Anemia was defined according to WHO criteria.Linear regression models were used to test the relationship between IGF-1 and Hb and between IGF-1 and anemia in model 1 (adjusted for age) and in model 2 adjusted for age,T, erythropoietin,IL-6,BMI, energy intake,vitamin B-12, ferritin, folic acid, iron and liver function.Logistic regression models adjusted for baseline Hb levels were used to test the relationship between IGF-1 and anemia after 3 years, in not anemic patients.[br][bold]Results. [/bold]The mean age was 74.7[plusmn]7.0 (years [plusmn] SD) in men and 76.5[plusmn]7.7 in women. 45 men (11.2%) and 71 women (13.2%) were anemic.IGF-1 levels (ng/ml) were lower in anemic men (100.3[plusmn]54.2) and women (89.1 [plusmn] 48)compared to non anemic men (129.7[plusmn]55.8) and women (109.6 [plusmn] 51.7).The difference in IGF-1 levels between the two groups was significant in men (p = 0.0009) and women (p = 0.002)and almost significant after adjustment for age in men (p = 0.08) and women (p=0.16).After adjustment for age, IGF-1 was positively associated with Hb in men ([beta][plusmn]SE 0.59 [plusmn] 0.14, p[lt]0.0001)and women([beta][plusmn]SE 0.16 [plusmn] 0.08, p=0.048).After further adjustment for T and other confounders the relationship between IGF-1 and Hb was still significant in men ([beta][plusmn]SE 0.47 [plusmn] 0.12, p[lt]0.0001), but not in women ([beta][plusmn]SE 0.06 [plusmn] 0.07, p=0.39).IGF-1 levels were not independent predictors of anemia in men and women.[br][bold]Conclusions.[/bold]IGF-1 is positively and independently associated with Hb in older men and women. Anemic older patients, especially men, have lower levels of IGF-1 independent of age and other confounders. However IGF-1 is not an independent predictor of anemia in both men and women.[br][br]Sources of Research Support: 1.Makipour S, Kanapuru B, Ershler WB. Unexplained anemia in the elderly. Semin Hematol. 2008 Oct;45(4):250-4. Review. 2.Ferrucci L, Maggio M, Bandinelli S, Basaria S, Lauretani F, Ble A, Valenti G, Ershler WB, Guralnik JM, Longo DL. Low testosterone levels and the risk of anemia in older men and women. Arch Intern Med. 2006 Jul 10;166(13):1380-8. 3.Shahani S, Braga-Basaria M, Maggio M, Basaria S. Androgens and erythropoiesis: past and present. J Endocrinol Invest. 2009 Sep;32(8):704-16. Epub 2009 Apr 7. Review. 4.Succurro E, Arturi F, Caruso V, Rudi S, Sciacqua A, Andreozzi F, Hribal ML, Perticone F, Sesti G. Low insulin-like growth factor-1 levels are associated with anaemia in adult non-diabetic subjects.Thromb Haemost. 2011 Feb 1;105(2):365-70. Epub 2010 Nov 5.[br][br]Nothing to Disclose: MM, FL, CC, SP, FR, SM, GC, AV, SM, SB, LF, GPC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1084 320 2540 MON-672 PO18-02 Monday 2209 2012


2205 ENDO12L_MON-673 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) The Severity of Short Stature in Two Siblings Heterozygous for a Novel Bioinactive Growth Hormone Mutant (GH-P59S) Suggests That the Mutant Affects GH Sensitivity to Endogenous Wild-Type GH Vibor Petkovic, Maria Consolata Miletta, Andree Eble, Amit Pandey, Annemieke Boot, Monique Losekoot, Christa Emma Fluck, Jan-Maarten Wit, Primus Eugen Mullis University Children[apos]s Hospital, Bern, Switzerland; University Medical Centar Griningen, Groningen, Netherlands; Leiden University Medical Center, Leiden, Netherlands; Leiden University Medical Center, Leiden, Netherlands Short stature caused by biologically inactive growth hormone (GH) is clinically characterized by lack of GH action despite normal-high secretion of GH, pathologically low IGF-1 concentrations and marked catch-up growth on GH-replacement therapy.[br]The index patients, adopted siblings (girl and a boy) of unknown clinical family history, were referred for assessment of short stature (-4.5 and -5.6 SDS). They had delayed bone ages (6.8 yrs and 4.5 yrs at chronological ages of 10 yrs and 8.1 yrs), normal GH peaks at stimulation tests (17 ng/ml and 11 ng/ml) and severely reduced IGF-1 concentrations (-3.5 and -4.0 SDS), respectively. Genetic analysis of the GH-1 gene showed a heterozygous P59S mutation in both patients. Based on the position of P59S mutation that lies within a patch of residues composing the GH binding site 1 for GHR, we performed an analysis of GH-P59S binding to GHR by in silico mutagenesis and molecular dynamics simulations, which suggested possible problems in correct binding of GH-P59S to the GHR. In vitro functional characterization of the mutant assessed through studies of GHR binding and activation of Jak2/Stat5 signaling pathway revealed reduced binding affinity of GH-P59S for GHR (IC[sub]50[/sub] 30 ng/ml) when compared to the wt-GH (IC[sub]50[/sub] 11.8 ng/ml) while a significantly decreased ability to activate the Jak2/Stat5 signaling pathway was observed at physiological concentrations of 25-100 ng/ml.[br]The clinical and biochemical data of our patients support the diagnosis of bioinactive GH syndrome, but the severity of the growth failure observed in these patients cannot solely be explained by the moderate effects of GH-P59S on GH binding and signaling. However, at physiologically normal endogenous secretion of GH, the mutant GH variant is affecting the wt-GH function on GHR binding as well as signalling altering its responsiveness to either wt-GH and/or rhGH treatment, which is important for normal growth.[br][br]Nothing to Disclose: VP, MCM, AE, AP, AB, ML, CEF, J-MW, PEM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 498 320 2541 MON-673 PO18-02 Monday 2210 2012


2206 ENDO12L_MON-674 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Sodium Butyrate Treatment Increases Growth Hormone (GH) Secretion in Autosomal Dominant GH Deficiency Caused by the Splice Site Mutations Maria Consolata Miletta, Vibor Petkovic, Andree Eble, Christa Emma Fluck, Primus Eugen Mullis University Children[apos]s Hospital, Bern, Switzerland Autosomal dominant form of isolated growth hormone deficiency type II (IGHD II) is primarily a splicing disorder and results from heterozygous splice site mutations that weaken recognition of exon 3 splice sites leading to aberrant splicing of [italic]GH-1[/italic] transcripts. This leads to an increasing production of a 17.5-kDa GH isoform which than exhibits a dominant-negative effect on the secretion of 22-kDa isoform (active form). [italic]In vitro[/italic] and transgenic animal data suggest that the onset and severity of the IGHD II phenotype reflect a threshold and a dose dependency effect depending of the amount of 17.5-kDa relative to 22-kDa isoform.[br]Sodium Butyrate (NaB), an histone deacetylase inhibitor, has been considered as a new promising agent for cancer therapy due to the absence of its toxicity and to multiple effects it exerts on cultured mammalian cells including inhibition of proliferation, induction of cell differentiation and apoptosis and induction/repression of gene expression. Recently, NaB was also found to act as a splicing modulator and its potential role as drug has been extended to splicing disorders.[br]In order to investigate the potential of NaB treatment to recover the normal GH secretion in an [italic]in vitro[/italic] model of IGHD II, we transfected rat pituitary cell line stably expressing hGHRHR (GC-GHRHR) cells with [italic]wt[/italic]-GH and/or different GH-splice site mutants (IVS+2, IVS+6 and ISE+28), stimulated with GHRH (10nM) and/or additionally treated with NaB (5mM) for 24h.[br]Treatment with NaB significantly increased extracellular GH secretion, in GHRH-stimulated and in non-stimulated conditions. Furthermore, in GHRH-stimulated conditions NaB works like a booster for GHRH, enhancing its effect on GH secretion. Moreover, NaB decreased 17.5-kDa/22-kDa GH ratio, the key determinant of IGHD II, both at transcriptional (analyzed by qRT-PCR) and at protein level (western blot).[br]At present, patients suffering from IGHD II are treated with costly daily injections of recombinant human GH (rhGH) in order to maintain normal growth and other anabolic effects; however, it does not prevent the development of other pituitary hormone deficiencies that eventually complicate the clinical picture in many of these patients. Thus, there is a need for alternative approaches to IGHD II therapy. Taken together, NaB might be a good candidate to test a new therapeutic approach to treat patients suffering from IGHD II.[br][br]Nothing to Disclose: MCM, VP, AE, CEF, PEM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 463 320 2542 MON-674 PO18-02 Monday 2211 2012


2207 ENDO12L_MON-675 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) The Onset of Inhibition of Growth Hormone Secretion Produced by SXN101959 [italic]In Vivo[/italic] Occurs within 24 Hours and Involves a Transient Stimulation of Growth Hormone Secretion Elaine A Harper, Alberto Martinez, Sandra Marlin, Julien Browne, Julie Rainard, Matthew Beard, Elena Fonfria, Elizabeth Marks, Sarah Donald Syntaxin Ltd, Abingdon, UK SXN101959 is a Targeted Secretion Inhibitor (TSI) targeted to somatotroph pituitary cells through a modified version of growth hormone releasing hormone (GHRH) and in rats produces a dose-dependent inhibition of pulsatile GH secretion following i.v. administration. The mechanism of action of SXN101959 is believed to involve binding and activation of GHRH receptors, receptor mediated internalisation, endosomal escape of the endopeptidase domain and cleavage of VAMP proteins required for vesicular secretion of GH. The aim of this study was establish the time to onset of action of SXN101959 [italic]in vivo[/italic] and further explore its mechanism of action (MOA) [italic]in vitro [/italic]in target cells. In light of this, the ability of SXN101959 to activate GHRH receptors in rat primary pituitary cells was monitored by measurement of cAMP accumulation and its ability to evoke GH secretion from these cells was also measured. SXN101959 treatment resulted in both a concentration-dependent increase in intracellular cAMP accumulation and secreted GH. To understand the time to onset of action [italic]in vivo[/italic], five groups of eight femoral vein and artery canulated Sprague Dawley rats were dosed (i.v.) with SXN101959 (0.3 mg/kg). Blood samples were collected from study group one every 15 min 5 h prior to dosing and then for 6 h post dosing. On days 2 to 5, bloods were collected from the other study groups starting at 24, 48, 72 and 96 hours after dosing. Plasma GH levels were measured using a Perkin Elmer AlphaLISA assay. Prior to dosing, rats in the first group exhibited pulsatile GH secretion with mean total plasma GH (AUC) levels of 104.2[plusmn]14.9 ng/ml over a 5 h period. 15-30 min post dosing, there was a 20-fold increase in plasma GH (AUC = 1995[plusmn]192 ng/ml over the next 6 h). 24 h post dosing, there was a decrease in plasma GH AUC levels to 69.1[plusmn]13.9 ng/ml and thereafter there was a significant and continued exponential decrease in plasma GH, with an approximate halving every 24 h (AUC ng/ml 48 h =32.5[plusmn]5.5, 72 h = 14.5[plusmn]6.3 and 96 hours 6.5[plusmn]1.4, respectively; p[lt]0.05, one way ANOVA and Bonferroni Multiple Comparison Post Test; all data are mean [plusmn] s.e.mean). The data suggest that the MOA of SXN101959 involves a transient stimulation of GH secretion through activation of pituitary GHRH receptors. This is followed by an inhibitory effect, mediated by SNARE cleavage that starts to take effect within 24 h.[br][br]Disclosures: AM: Employee, SYNTAXIN. Nothing to Disclose: EAH, SM, JB, JR, MB, EF, EM, SD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1628 320 2543 MON-675 PO18-02 Monday 2212 2012


2208 ENDO12L_MON-676 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Perturbations of the IGF System Following Commencement of Statin Therapy in a Type 2 Diabetes Population with Early Nephropathy Ram Prakash Narayanan, Matt Gittins, Kirk W Siddals, Robert L Oliver, Julie E Hudson, Anne White, Paul N Durrington, Robert R Davies, Martin K Rutter, John Martin Gibson The University of Manchester, Salford, UK; The University of Manchester, Manchester, UK; The University of Manchester, Manchester, UK; Manchester Royal Infirmary, Manchester, UK Statins have a major role in vascular prevention, but these benefits may be offset by a small increase in diabetes risk through unknown mechanisms. Since insulin-like growth factors and their binding proteins influence the development of type 2 diabetes, we compared the effects of low-dose versus high-dose atorvastatin on the IGF system in patients with type 2 diabetes randomised to receive either 10 mg or 80 mg of atorvastatin enrolled in the PANDA trial1 (N=119; mean (SD): baseline age 64 (10) years; 83% male; HbA1c 61 (10) mmol[frasl]mol, blood pressure 131/73 mmHg; 80% received renin-angiotensin system blockade, all had microalbuminuria or proteinuria).[br]We measured plasma IGF-I, IGFBP-1 and IGFBP-3, and serum IGF-II using ELISA-based methods at baseline, 6 months and 12 months. IGFBP-1 values were log-transformed. A mixed effects longitudinal model was used to study differences in effects of 10mg and 80mg atorvastatin on concentrations of proteins, and on IGF-I:IGFBP-1 and IGF-I:IGFBP-3 ratios. All changes from baseline were adjusted for baseline level, age sex, BMI, and prior statin and insulin therapy.[br]Concentrations of IGF-I, IGF-II and IGFBP-3 reduced from baseline with high and low dose therapy. The adjusted mean (95% CI) between-group (10mg vs 80mg atorvastatin) difference for change in IGF proteins was not significant for: IGF-I: -3 (-21 to 14) ng/ml; IGF-II: -23 (-65 to 18) ng/ml; and IGFBP-3: -0.34 (-0.71 to 0.03) [micro]g/ml; negative values indicating numerically greater lowering with high-dose).[br]Concentrations of IGFBP-1 were reduced in patients receiving high-dose, but they increased from baseline with low-dose therapy. The adjusted mean (95% CI) between-group difference for the change in log IGFBP-1 was significant -0.41 (-0.69 to 0-0.13, p=0.004). The corresponding change in IGF-I/IGFBP-1 ratio: 6.5 (1.3 to 11.7), p=0.015 implies altered IGF bioavailability. Changes in all IGF proteins were not correlated with changes in HbA1c.[br]This study reports the longitudinal effects of different doses of statins on four important IGF proteins in type 2 diabetes. The differential effects of low vs high dose statins on IGFBP-1 concentrations are novel and intriguing. IGFBP-1 exerts dynamic effects on IGF-I bioactivity and low IGFBP-1 is predictive of incident diabetes. It is plausible that long term treatment with high vs low dose of atorvastatin may differentially impact IGF bioavailability and part-mediate the incident diabetes reported in recent statin trials.[br][br]1)Rutter MK et al.,Diabet Med. 2011 Jan;28(1):100-8.[br][br]Sources of Research Support: This study was funded by an unrestricted grant from Pfizer that markets atorvastatin.[br][br]Nothing to Disclose: RPN, MG, KWS, RLO, JEH, AW, PND, RRD, MKR, JMG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1631 320 2544 MON-676 PO18-02 Monday 2213 2012


2209 ENDO12L_MON-677 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) The Limited Screening Value of Insulin-Like Growth Factor-I as a Marker for Alterations in Body Composition in Very-Long-Term Adult Survivors of Childhood Cancer Karin Blijdorp, Marry N van den Heuvel-Eibrink, Rob Pieters, A J van der Lely, Sebastian JCMM Neggers Erasmus University Medical Centre Rotterdam, Rotterdam, Netherlands; Erasmus University Medical Centre Rotterdam-Sophia Children[apos]s Hospital, Rotterdam, Rotterdam, Netherlands Introduction[br]The clinical relevance of low IGF-I levels, caused by cranial radiotherapy, in adult childhood cancer survivors has not been studied extensively. We evaluated whether IGF-I is a useful marker for altered body composition and growth hormone deficiency in this group.[br]Methods[br]We analyzed retrospective data from 610 adult childhood cancer survivors, retrieved from the late effects clinic. Median age at diagnosis was 6 years (interquartile range 3-11) and follow-up time was 18 years (13-24). We assessed IGF-I standard deviation scores (SDS), anthropometrical measures, growth hormone stimulation tests in patients with clinical signs of GHD and measures of body composition (assessed by dual X-ray absorptiometry, Lunar Prodigy).[br]Results[br]In 58 cranially irradiated acute leukemia survivors (25 Gy (24-25)) and 56 locally irradiated brain tumor survivors (42 Gy (35-54)) we found significantly lower IGF-I SDS (p[lt]0.001), lower height SDS (p[lt]0.001), higher body mass index (p=0.01), higher waist-hip ratio (p=0.001), higher total fat percentage SDS (p[lt]0.001) and lower lean body mass SDS (p[lt]0.001), as compared to 452 not cranially irradiated survivors. IGF-I showed a weak inverse correlation with BMI (r=-0.12, p=0.04), waist hip ratio (r=-0.15, p=0.01), total fat percentage (r=-0.14, p=0.02) and a positive correlation with lean body mass (r=0.15, p=0.01). In patients with low IGF-I levels, IGF-I did not significantly differ between subjects with and without GHD as determined by GH-stimulation testing (p = 0.39).[br]Conclusion This study shows that IGF-I has limited value as a marker for alterations in body composition in adult childhood cancer survivors.[br][br]Nothing to Disclose: KB, MNvdH-E, RP, AJvdL, SJCMMN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 75 320 2545 MON-677 PO18-02 Monday 2214 2012


2210 ENDO12L_MON-678 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) The Relationship between Low Insulin-Like Growth Factor-1 (IGF-1) Levels, Exercise Capacity, Muscle Strength and Vitality in Adult Survivors of Acute Lymphoblastic Leukemia (ALL) Treated with Cranial Radiotherapy (CRT) Wassim Chemaitilly, Melissa M Hudson, Sujuan Huang, Webb A Smith, Deo Kumar Srivastava, Nicole Barnes, Ching-Hon Pui, Larry E Kun, Charles A Sklar, Leslie L Robison, Kirsten K Ness St Jude Children[apos]s Research Hospital, Memphis, TN; St Jude Children[apos]s Research Hospital, Memphis, TN; St Jude Children[apos]s Research Hospital, Memphis, TN; St Jude Children[apos]s Research Hospital, Memphis, TN; St Jude Children[apos]s Research Hospital, Memphis, TN; St Jude Children[apos]s Research Hospital, Memphis, TN; Memorial Sloan Kettering Cancer Center, New York, NY [bold]Background[/bold][br]Long-term survivors of childhood ALL have reduced physical fitness when compared to age-matched individuals. Those treated with CRT are also at risk of growth hormone deficiency (GHD). The possible contribution of GHD to reduced physical fitness in this vulnerable population is unknown.[br][bold]Aim[/bold][br]To evaluate the association between IGF-1 plasma levels and exercise capacity, muscle strength and vitality in a group of adult survivors of childhood ALL treated with CRT.[br][bold]Methods[/bold][br]Subjects were enrolled in the St Jude Lifetime protocol. Investigations necessary for the present study were completed by 394 subjects (200 males) aged 18 to 54 (34.8[plusmn]6.8) years, who were[lt]21 years (94.2% [lt]15years) at ALL diagnosis. CRT doses to the hypothalamus/pituitary were[gt]18 Gy in 58.1%. Those on GH therapy were excluded. Exercise capacity was measured using peak oxygen consumption (VO[sub]2[/sub]peak) and 6 minute walk distance. Isokinetic knee extension and isometric hand grip strengths were used for measuring peak muscle strength; the SF-36 score for assessing vitality. IGF-1 levels were expressed in z-score (zs). Values are expressed as the mean [plusmn] standard deviations (SD).[br][bold]Results[/bold][br]There were no significant differences between subjects with IGF-1[lt]-2zs (n=188) and those with an IGF-1[ge]-2zs in V[sub]O2[/sub]peak (25.8[plusmn]6.9 vs. 26.0[plusmn]5.9 ml/kg/min, p = 0.81). Subjects with IGF-1[lt]-2zs had shorter 6-minute walk distance (537.9[plusmn]109.3 vs.570.6[plusmn]98.5 m,p [lt]0.001), lower knee extension (136.1[plusmn]50.4 vs.159.6[plusmn]52.8 Nm/kg, p[lt]0.001) and lower hand grip (35.0[plusmn]12.9 vs. 40.4[plusmn]14.2 kg, p[lt]0.01) strengths, and a lower SF-36 score (46.4 [plusmn]11.2 vs. 49.4[plusmn]10.7, p[lt]0.01). After adjustment for sex, age, age at ALL diagnosis, BMI, tobacco/alcohol use, cardiac and pulmonary dysfunctions, the multivariable analysis did not identify an independent effect of IGF-1[lt]-2zs on V[sub]O2[/sub]peak (p=0.57), 6-minute walk distance (p=0.45), knee extension (p=0.16) or hand grip (p=0.08) strengths, or the SF-36 score (p=0.06).[br][bold]Conclusions[/bold][br]Adult survivors of childhood ALL treated with CRT with IGF-1 [lt]-2zs have decreased muscle strength and vitality when compared to others. The multivariate analysis did not identify IGF-1 [lt]-2zs as having an independent effect on these parameters after adjusting for demographic, treatment and lifestyle related risk factors which impact may hence override the effect of GHD. The amenability of physical fitness markers for improvement in adult survivors of childhood ALL with GHD deserves further investigation.[br][br]Sources of Research Support: American Lebanese Syrian Associated Charities.[br][br]Nothing to Disclose: WC, MMH, SH, WAS, DKS, NB, C-HP, LEK, CAS, LLR, KKN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 761 320 2546 MON-678 PO18-02 Monday 2215 2012


2211 ENDO12L_MON-679 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Ventilatory Equivalents Improve after Growth Hormone Replacement in Growth Hormone Deficient Patients Recovering from Traumatic Brain Injury Kurt Mossberg, Astrid Horstman, William Durham, Charles Gilkison, Brent Masel, Randall Urban University of Texas Medical Branch, Galveston, TX; University of Texas Medical Branch, Galveston, TX; Transitional Learning Center, Galveston, TX [bold]Background and Purpose: [/bold]Growth hormone (GH) deficiency is known to occur in approximately 20% of individuals who suffer from a moderate to severe traumatic brain injury (TBI). Our objective was to assess the effects of GH replacement on efficiency of breathing in individuals who sustained a TBI as adults.[br][bold]Subjects: [/bold]Twenty individuals who sustained a TBI at least 12 months prior to study enrollment were identified as GH deficient by glucagon stimulation testing (maximum GH response 8ng/mL or less). Subjects were then randomized into placebo (n=10) or recombinant human GH (rhGH) (n=10) groups.[br][bold]Procedures[/bold]: Subjects had peak cardiorespiratory capacity including pulmonary ventilation, oxygen consumption and carbon dioxide production determined by an automated metabolic cart during a graded treadmill exercise test. Heart rate was monitored by ECG. Initial walking speed was adjusted in the first 2 minutes of the test to accommodate for TBI induced gait impairments. Speed was then held constant while incline was increased 2% each minute until exhaustion. Tests were performed at baseline and after 12 months. After baseline testing, subjects injected either placebo or active drug subcutaneously daily for 1 year.[br][bold]Data Analysis:[/bold] Two-way analyses of variance were carried out on all cardiorespiratory variables (main effects: time and group). All analyses were carried out at [alpha] [lt] 0.05.[br][bold]Results: [/bold]No significant main effects or interactions were found for the cardiorespiratory variables. Large coefficients of variation were observed and were consistent with the variability in physical function (gait impairments) between subjects. No interactions were found between time and group. Ventilatory equivalents for carbon dioxide (V[sub]E[/sub]/VCO[sub]2[/sub]) and oxygen V[sub]E[/sub]/VO[sub]2[/sub]) showed the greatest differences between groups.[br][bold]Discussion and Conclusion: [/bold]The ventilatory equivalent is an estimate of efficiency of breathing. The lower the ratio of total air moved by the lungs per minute vs. gas exchanged equates to less ventilatory muscle work. Thus, less effort is required to exchange CO[sub]2[/sub] and O[sub]2[/sub] in the lungs. Subjects who received rhGH replacement tended to have improved breathing efficiency compared to those who injected placebo. Efficiency of breathing may play a role in the subjective complaints of fatigue observed in patients recovering from TBI.[br][br]Nothing to Disclose: KM, AH, WD, CG, BM, RU 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1547 320 2547 MON-679 PO18-02 Monday 2216 2012


2212 ENDO12L_MON-680 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Klotho as a Biomarker for Growth Hormone Deficiency in Traumatic Brain Injury Larry Denner, Sigmund J Haidacher, Charles Gilkison, Brent E Masel, Randall J Urban University of Texas Medical Branch, Galveston, TX; University of Texas Medical Branch, Galveston, TX; Transitional Learning Center, Galveston, TX Traumatic brain injury (TBI) often causes hypopituitarism as a result of growth hormone deficiency (GHD). This is associated with multiple physical, metabolic, and neuropsychological problems including diminished lean body mass, disrupted lipoprotein and carbohydrate metabolism, reduced bone mineral density and impaired cardiac function, as well as declines in cognitive functioning, fatigue, and diminished quality of life. We previously showed that early diagnosis of GHD and subsequent GH replacement therapy has significant benefits to recovery of patients with TBI. Nonetheless, diagnosis of patients with GHD can take considerable time in the post-injury period when early intervention is preferable. Furthermore, improvement in cognitive function is a long process, and evaluation of success is incremental and challenging. Thus, biomarkers for diagnosis and management of GHD in TBI patients can offer a solution to a large unmet medical need.[br]To meet this need, we are evaluating Klotho as a potential biomarker for extent of injury and prediction of response to therapy in TBI. Klotho is an important mediator of responses to stress and disease. It is a [sim]130kDa transmembrane protein that is cleaved by extracellular proteases to release a [sim]72kDa fragment that is found in blood, urine, and cerebral spinal fluid in many pathological processes such as diabetes. We developed methods to measure Klotho in serum of TBI patients using an Agilent 3560 high resolution triple quadrupole mass spectrometer to develop multiple reaction monitoring assays (MRMs). We use 5 tryptic peptides from Klotho with 1-3 fragments per peptide for the equivalent of using 5-15 unique antibodies to measure a protein as might be performed in an ELISA. Such a large number of measures, combined with stable heavy isotope-labelled internal standards, allows absolute quantification of Klotho in serum at levels above 25 attomoles (25 x 10-[sup]18[/sup] moles). Typically, 5 ul of patient serum is enriched for Klotho using cobalt metal affinity chromatography. After trypsinization, 1 ug of total peptides are mixed with internal standards and applied to the mass spectrometer with a total run time of 35 minutes per sample. Absolute quantification is performed automatically compared to the internal standards using instrument software. This assay is now being used to determine whether Klotho can be a biomarker for early diagnosis and management of TBI to bring improved quality of life to these patients.[br][br]Sources of Research Support: The NIH-National Institute of Child Health and Human Development (R01-HD044566) and the Miriam and Emmett McCoy Foundation.[br][br]Nothing to Disclose: LD, SJH, CG, BEM, RJU 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2049 320 2548 MON-680 PO18-02 Monday 2217 2012


2213 ENDO12L_MON-681 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Acute Response of Growth Hormone to Peak Treadmill Exercise in Patients Recovering from Traumatic Brain Injury William Amonette, Patricia Lea, Brent Masel, Melinda Sheffield-Moore, Randall Urban, Kurt Mossberg University of Texas Medical Branch, Galveston, TX; University of Texas Medical Branch, Galveston, TX; Transitional Learning Center, Galveston, TX; University of Texas Medical Branch, Galveston, TX [bold]Introduction:[/bold] Approximately 20% of patients recovering from a traumatic brain injury (TBI) are growth hormone (GH) deficient (glucagon stimulation test [lt]3.0 ng/dL) and GH deficiency is related to lower peak aerobic capacities in TBI. Vigorous exercise is a potent stimulus for GH release which is essential for substrate mobilization. Therefore, we hypothesized that a blunted exercise-induced GH response could be responsible, in part, for reduced peak aerobic capacities observed in patients with a TBI. The purpose of this study was to compare the post-exercise GH levels of patients with a TBI to healthy controls (CON).[br][bold]Methods:[/bold] Eight patients with a TBI (TBI; 7m, 1f; 31.3[plusmn]2.0y; 177.8[plusmn]3.6cm; 96.1[plusmn]5.5kg) and 8 age- and gender-matched healthy controls (CON; 7m, 1f; 31.3[plusmn]2.0y; 177.8[plusmn]3.6cm; 96.1[plusmn]5.5kg) volunteered. Subjects completed a maximal treadmill graded exercise test to volitional failure. Venous blood was sampled 7 times prior to and after exercise in 10-minute intervals, and serum GH levels were measured from these 14 time-points. Integrated GH values (AUC) were calculated for the 30-minute interval prior to and immediately after exercise (PRE and POST 0-30), and again 30-60 minutes after exercise (POST 30-60). A two-factor ANOVA was used to compare the main effects of group (TBI vs. CON) and time (PRE, POST 0-30; POST 30-60). Pair-wise comparisons were accomplished using Tukey[apos]s HSD with alpha set at [italic]p[/italic][le]0.05.[br][bold]Results:[/bold] A significant main effect was detected for time (F = 5.9; [italic]p[/italic] [lt] 0.001) but not group (F = 0.005; p = 0.94). GH was elevated in both groups during the immediate post exercise period (POST 0-30: TBI [Delta] 63.5[plusmn]29.1 and CON [Delta] 48.9[plusmn]18.5 ng/dL/min) with no between group differences. GH at POST 30-60 were near PRE levels in both groups (TBI: [Delta] 27.5[plusmn]11.6 and CON [Delta] 44.3[plusmn]21.4 ng/dL/min). Individual data revealed that 5 patients with TBI had minimal or no GH release ([lt]1.0 ng/dL) at any time after exercise.[br][bold]Conclusions:[/bold] Exercise-stimulated GH release in the post-exercise period appears similar in TBI and controls. However, exercise-stimulated GH release is not stimulated consistently in all patients with TBI and is significantly blunted or absent in over half of those with TBI. Future research is warranted to determine if patients with a TBI who exhibit minimal GH elevation after exercise are clinically GH-deficient, and if this blunted response is mechanistically related to low aerobic fitness levels previously described in patients with a TBI.[br][br]Nothing to Disclose: WA, PL, BM, MS-M, RU, KM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1508 320 2549 MON-681 PO18-02 Monday 2218 2012


2214 ENDO12L_MON-682 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) IGF-I Bioactivity Might Better Reflect Quality of Life Than Total IGF-I in GH-Deficient Patients during GH Treatment Aimee J Varewijck, Steven WJ Lamberts, Sebastian JCMM Neggers, Leo J Hofland, Joseph AMJL Janssen Erasmus Medical Center, Rotterdam, Netherlands [bold]Context[/bold] GH therapy of GH deficient adults has been shown to improve quality of life (QoL) in some, but not all studies. No correlation has been found between the degree of improvement of QoL and total IGF-I during GH therapy. It has been suggested that IGF-I bioactivity is more sensitive than total IGF-I to detect differences in clinical state.[br][bold]Aim[/bold] To investigate the value of IGF-I bioactivity in monitoring QoL in patients with GHD treated with GH for 12 months.[br][bold]Methods[/bold] 106 patients (54 m; 52 f) with GHD were included; 22 were GH-naive, 84 were already on GH treatment and discontinued therapy 4 weeks before establishing baseline values. IGF-I bioactivity was determined by the IGF-I KIRA assay, total IGF-I by immunoassay (Immulite) in fasting blood samples collected at baseline and after 12 months of GH therapy. GH doses were titrated to achieve total IGF-I within the normal range. QoL was measured by the disease-specific Question on Life Satisfaction Hypopituitarism Module (QLS-H) and by the general SF-36 questionnaire at baseline and 12 months thereafter.[br][bold]Results[/bold] Total IGF-I increased from 8.2[plusmn]0.5 nmol/l (Z-score -2.2SD) at baseline to 14.9[plusmn]0.7 nmol/l (Z-score -0.6SD) at 12 months. IGF-I bioactivity increased from 116[plusmn]6 pmol/l (Z-score -2.6SD) to 179[plusmn]11 pmol/l (Z-score -1.8SD). At 12 months the mean GH dose was 0.20 mg/day (range: 0.05-2.1).[br]QLS-H did not change over 12 months (-0.68[plusmn]0.3 SD vs. -0.70[plusmn]0.17 SD, p=0.83). After 12 months, the mental component summery component of the SF-36 questionnaire had increased significantly (68[plusmn]2% vs. 72[plusmn]2%, p=0.03), while the physical summery component had not (67[plusmn]2% vs. 68[plusmn]2%, p=0.70).[br]After 12 months, both total IGF-I and IGF-I bioactivity were (age-adjusted) positively related to the physical summery component (r=0.30, p=0.009; r=0.40, p=0.001, respectively) and to the mental component summery component (r=0.28, p=0.02; r=0.35, p=0.004, respectively). Interestingly, IGF-I bioactivity was (age-adjusted) positively related to QLS-H (r=0.26, p=0.04), while total IGF-I was not (r=0.04, p=0.74).[br][bold]Conclusion[/bold] Only mental health increased during 12 months of GH therapy in GH deficient patients. Although IGF-I bioactivity was still subnormal after GH treatment, there was a positive relationship with the disease specific QLS-H module, while this relationship was absent for total IGF-I. This suggests that IGF-I bioactivity might better reflect QoL than total IGF-I in GH deficient patients during GH treatment.[br][br]Sources of Research Support: J.A.M.J.L. Janssen has received an unrestricted grant from Novo Nordisk A/S (Alphen aan de Rijn, the Netherlands).[br][br]Nothing to Disclose: AJV, SWJL, SJCMMN, LJH, JAMJLJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1479 320 2550 MON-682 PO18-02 Monday 2219 2012


2215 ENDO12L_MON-683 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Pharmacological Characterization of NNC0195-0092, a Long-Acting Growth Hormone Derivative Peter Thygesen, Henrik Sune Andersen, Christian Rischel, Johannes Joseph Fels, Nils Langeland Johansen Novo Nordisk A/S, Maaloev, Denmark; Novo Nordisk A/S, Maaloev, Denmark; Novo Nordisk A/S, Maaloev, Denmark; Novo Nordisk A/S, Maaloev, Denmark; Novo Nordisk A/S, Maaloev, Denmark Growth hormone (GH) is an important regulator of longitudinal growth in children and metabolism in adults. GH supplementation is used for the treatment of various deficiency disorders in both children and adults. GH treatment is safe and well tolerated. However, its clinical use is limited to daily subcutaneous injections which poses a challenge for both the patients and the physicians and has prompted research into alternative approaches to GH treatment.[br]NNC0195-0092 is a hGH derivative conjugated with an albumin binding moiety. The in vivo pharmacodynamic and pharmacokinetic properties of NNC195-0092 have been studied in hypophysectomised rats, mini-pigs and cynomolgus monkeys.[br]NNC0195-0092 induced a dose-dependent increase in body weight of hypophysectomised rats and increase in plasma IGF-I levels in rats, mini-pigs, and cynomolgus monkeys. Multiple once weekly dosing of hypophysectomised rats with NNC195-0092 induced a step-wise increase in body weight with an initial linear growth the first 3-4 days in each dosing interval. During the 28 days study period with 4 once weekly doses of NNC195-0092, body weight was increased by 52% compared to a vehicle control group. The IGF-I levels increased after each dose and returned to the same trough level in each dosing interval. Significant increases in body lean mass, tibia bone mineral content, and tibia cortical thickness were observed together with a decrease in body fat mass compared to a vehicle control group.[br]Pharmacokinetics in all three species has indicated a 1. order absorption from the subcutaneous tissue after a s.c. injection and a non-linear elimination. Apparent terminal half-lives of 5-6 hours in rats, 10-12 hours in mini-pigs, and 17-20 hours in monkeys have been observed. The bioavailability was estimated to 39% in rats, 36 % in mini-pigs, and 69 % in monkeys.[br]In conclusion, once weekly s.c. administration of NNC0195-0092 exhibit pharmacokinetic and pharmacodynamic properties indicating that NNC195-0092 is a potential once weekly growth hormone candidate.[br][br]Disclosures: PT: Employee, Novo Nordisk. HSA: Grant Review Panel, Novo Nordisk. CR: Investigator, Novo Nordisk. JJF: Grant Review Panel, Novo Nordisk. NLJ: Employee, Novo Nordisk. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1276 320 2551 MON-683 PO18-02 Monday 2220 2012


2216 ENDO12L_MON-684 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Development of Long-Acting Human Growth Hormone Suitable for Once-a-Week Dosing Joselyn Del Rosario, Moorthy Palanki, Zemeda Ainekulu, Michael Bacica, Lioudmila Campbell, Danielle Dettling, Kathryn Gropp, Mark Horn, Annette Sievers, Nebojsa Stankovic, Dean Wilkie, Abhijit Bhat, Rodney Lappe, Gary Woodnutt, Gang Chen Pfizer Worldwide Research and Development, San Diego, CA; Pfizer Worldwide Research and Development, Chesterfield, MO; Pfizer Worldwide Research and Development, Groton, CT; Pfizer Worldwide Research and Development, Cambridge, MA Recombinant human growth hormone (hGH) is used to treat growth hormone deficiency in children and adults, however its very short circulating half-life of 20-30 minutes requires daily injection. A long acting growth hormone receptor (GHR) agonist with sufficient stability to be dosed weekly is a highly desirable alternative. Rapid degradation of hGH has thus far precluded development of a therapeutic molecule with a PK half-life greater than 20 hours. We developed a long-acting hGH analogue conjugated to an aldolase carrier antibody engineered in such a way that human growth hormone was irreversibly linked to one of the Fab arms. The resulting conjugated molecule, known as a CovX-Body, has the pharmacological properties of GH with the benefit of extended half-life. In our early studies, fusing hGH to a CovX-Body resulted in a 4-fold improvement of hGH half-life (from 30 minutes to 2 hours). We suspected that the clearance of hGH-CovX-Body was mediated by receptor internalization and theorized that reducing agonist potency while maintaining receptor signaling of a hGH derivative would slow hGHR-mediated internalization and thereby extend hGH half-life. We developed a hGH CovX-Body with reduced [italic]in vitro [/italic]activities in IM9 and NB2 cell-based assays with uncompromised GH efficacy [italic]in vivo[/italic]. Pharmacokinetic studies in rat showed that the half-life of the hGH CovX-Body was increased to 29 hours following a single SC dose (10 mg/kg). Furthermore, [italic]in vivo[/italic] studies in hypophysectomized rats demonstrated that weekly SC doses (1.5 mg/kg, 5 mg/kg, or 50 mg/kg) of the hGH CovX-Body increased body weight and tibial length significantly as compared to the untreated hypophysectomized rats. With its superior pharmacokinetic profile and comparable efficacy to wild-type growth hormone, the long-acting hGH-CovX-Body has potential to become a novel therapeutic.[br][br]Nothing to Disclose: JDR, MP, ZA, MB, LC, DD, KG, MH, AS, NS, DW, AB, RL, GW, GC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1791 320 2552 MON-684 PO18-02 Monday 2221 2012


2217 ENDO12L_MON-685 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) A Retrospective Study of Comorbidities in Acromegaly Patients in the U.S. Tanya M Burton, Elisabeth Le Nestour, Tim Bancroft, Francis DiDonato, Maureen Neary OptumInsight, Eden Prairie, MN; PharmaNet/i3, Nanterre, France; Novartis Pharmaceuticals Corporation, East Hanover, NJ [bold]OBJECTIVE[/bold]: Describe comorbidities of acromegaly patients in a real-world treatment setting.[br][bold]METHODS:[/bold] A retrospective claims study in a US commercial health plan analyzed adults with an acromegaly-related diagnosis, procedure, or medication between 07/2002-06/2010. Patients were observed for 6 months before their first acromegaly-related claim and until death or disenrollment. The index date was defined as the date of the first acromegaly-related claim. Patients were observed for 6 months before the index date (baseline) and from the index date until death or disenrollment (follow-up). Cohorts were defined by claims for an acromegaly-related surgery and/or medication on or after the index date: surgery only (surg-only), medications only (meds-only), surgery and medications (surg-meds), or neither surgery nor medications (none). Baseline (BL) and follow-up (FU) comorbidity rates were summarized descriptively.[br][bold]RESULTS[/bold]: Of 1,056 pts, mean (SD) age was 42 (17) years on first acromegaly claim, 49% were male, 15% had surg-only, 21% had meds-only, 14% had surg-meds, and 50% had none. Overall, proportions with comorbidities increased substantially between BL and FU. Notable differences included: hypertension (BL=23.2%, FU=42.0%), arthropathy/arthralgia/synovitis (BL=14.3%, FU=35.8%), diabetes (BL=14.4%, FU=26.8%), valvular heart disease (BL=2.8%, FU=12.9%), and cardiac dysrhythmia/arrhythmia (BL=4.9%, FU=13.7%). The two surgery cohorts had similar BL and FU comorbidity profiles, except surg-only had a higher BL proportion of nervous system and lipid metabolism disorders than surg-meds. The surg-only and surg-meds cohorts also had the same or higher proportion of comorbidities during BL and FU than meds-only and none cohorts, except that surg-only cohort had lower FU proportion of myopathy/myalgia than the none cohort. The meds-only and none cohorts had similar BL and FU comorbidity profiles.[br][bold]CONCLUSION[/bold]: In this study, acromegaly patients had a high comorbidity burden, with considerable increases in serious comorbidities (hypertension, diabetes, valvular heart disease, and cardiac dysrhythmia/arrhythmia) across time. While a large proportion of acromegaly patients received neither surgery nor medication, given the chronic nature of this condition, some patients may have received acromegaly treatment prior to the observation period. Results support the need for timely diagnosis and treatment of acromegaly.[br][br]Disclosures: MN: Employee, Novartis Pharmaceuticals. Nothing to Disclose: TMB, ELN, TB, FD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 906 320 2553 MON-685 PO18-02 Monday 2222 2012


2218 ENDO12L_MON-686 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Pharmacological Treatment of Acromegaly: A Retrospective Study in a Large U.S. Claims Database Tanya M Burton, Elisabeth Le Nestour, Tim Bancroft, Francis DiDonato, Maureen Neary OptumInsight, Eden Prairie, MN; PharmaNet/i3, Nanterre, France; Novartis Pharmaceuticals Corporation, East Hanover, NJ [bold]OBJECTIVE[/bold] Describe the pharmacological treatment patterns of acromegaly patients in a real-world treatment setting.[br][bold]METHODS[/bold]: A retrospective claims study in a US commercial health plan analyzed adults with an acromegaly-related diagnosis, procedure, or medication between 07/2002-06/2010. The index date was defined as the date of the first acromegaly-related claim. Patients were observed for 6 months before the index date (baseline) and from the index date until death or disenrollment (follow-up). Cohorts were defined by the first claim for an acromegaly-related medication on or after the index date.[br][bold]RESULTS: [/bold]Of the 1,056 acromegaly patients identified, 370 (35%) had a claim for an acromegaly-related medication during follow-up (FU). Patients with a medication most often started FU with cabergoline (CAB: 29%, 107 of 370), octreotide acetate (OCT_ACE: 28%), and bromocriptine (BROM: 21%), followed by octreotide LAR (OCT_LAR: 14%), pegvisomant (PEG: 5%), and lanreotide (LAN: 3%). Only 17% (64 of 370) had an acromegaly-related medication claim during baseline (BL). The most common BL claims were 56% for OCT_ACE (36 of 64), 30% for BROM, and 22% for CAB. There were no BL claims for lanreotide or pegvisomant. For patients who started with OCT_ACE, the most commonly added medications to FU treatment regimens were OCT_LAR (53%), PEG (16%), and CAB (16%); and patients who started with CAB, commonly added OCT_ACE (14%) and OCT_LAR (10%). Of the 686 patients with no claims for an acromegaly-related medication, 168 had evidence of an acromegaly-related procedure some time during the study period (e.g., surgery or radiotherapy). Overall, almost half (49%) of the acromegaly patients identified had no evidence of any acromegaly treatment, neither medical nor surgical, during the study period.[br][bold]Conclusion:[/bold] A large proportion of acromegaly patients were untreated during the observation period. While some of these patients may have been in remission or cured prior to study enrollment, some of them may have been under-treated. Given the life-long consequences and comorbidities associated with acromegaly, future studies need to evaluate possible reasons for under-treatment and increase awareness about this rare condition and its effective treatment options.[br][br]Disclosures: TMB: Researcher, Novartis Pharmaceuticals. ELN: Clinical Researcher, Novartis Pharmaceuticals. TB: Researcher, Novartis Pharmaceuticals. FD: Researcher, Novartis Pharmaceuticals. MN: Employee, Novartis Pharmaceuticals. 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1795 320 2554 MON-686 PO18-02 Monday 2223 2012


2219 ENDO12L_MON-687 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Comparison of SXN101959, a Targeted Secretion Inhibitor (TSI), with the Gold Standard Octreotide To Lower the GH/IGF-I Axis Activity: A New Approach for Acromegaly Emmanuel Somm, Alberto Martinez, Audrey Toulotte, Petra S Huppi, Richard Jones, Michel L Aubert University of Geneva School of Medicine, Geneva, Switzerland; Syntaxin Ltd, Abingdon, UK [underline]Introduction[/underline]: Adenomas within pituitary somatotrophs generate abnormally high GH levels resulting in acromegaly, an endocrine disease characterized by an aberrant growth pattern. SXN101742 and 101959 are TSIs designed to enter somatotrophs, cleave SNARE proteins thus preventing GH granules exocytosis. We previously showed that a single IV administration of SXN101742 led to specific dose-dependent decreases in GH and IGF-I plasma levels and gene expression in a juvenile rat model. The goal of the present study was to compare, in the same growing rat model, the efficacy of the somatostatin analog Octreotide with SXN101959, a new TSI, derived from SXN101742, and under development for clinical studies.[br][underline]Methods[/underline]: Treatment was initiated in male Sprague Dawley rats at 45 days of life to take advantage of the high GH/IGF-1 axis activity at this age. A single IV injection of SXN101959 (1mg/kg) was administered and compared to animals treated with a constant SC infusion of Octreotide (SMS201-995, 10[mu]g/kg/h). We monitored daily body weight gain and circulating GH and IGF-1 levels at 3-day intervals (with Millipore and IDS kits, respectively). Ten days after initiation of treatment, the rats were sacrificed and pituitary and hypothalamus, sampled for real time gene expression analysis (ABI).[br][underline]Results[/underline]: Body and pituitary weights were more reduced following the single bolus of SXN101959 than with the Octreotide infusion. Accordingly, both GH peaks and circulating IGF-I levels were suppressed more strongly in SXN101959-treated animals than in the Octreotide group. Gene expression for both pituitary GH and hypothalamic GHRH were markedly decreased in SXN101959-treated rats, consistent with a striking inhibition of GH synthesis. In contrast, no such effects were seen in rats treated with Octreotide for which pituitary GH mRNA and hypothalamic GHRH mRNAs remained normal.[br][underline]Conclusions[/underline]: A single bolus of SXN101959 exerted a more powerful inhibitory action than constant Octreotide infusion on the somatotropic axis in growing rats as seen 10 days after treatment initiation. Moreover, inhibition of GH synthesis was only seen with SXN101959, not with Octreotide. Such an inhibition of GH synthesis represents an important asset for long term inhibition of GH secretion. Taken together these data indicate that Syntaxin Botulinum toxin-derived TSI administered as single bolus, offer a greater potential than Octreotide to dampen the excessive GH/IGF-I axis activity.[br][br]Disclosures: AM: Employee, SYNTAXIN. RJ: Employee, SYNTAXIN. MLA: Consultant, SYNTAXIN. Nothing to Disclose: ES, AT, PSH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 371 320 2555 MON-687 PO18-02 Monday 2224 2012


2220 ENDO12L_MON-688 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) A Novel Formulation of Octreotide Enables Its Enteral Absorption and GH Inhibition in Animals Shmuel Tuvia, Dori Pelled, Alberto Kischitzky, Maya Levin-Arama, Karen Marom, Roni Mamluk Chiasma, Jerusalem, Israel [bold]Purpose[/bold]: Most therapeutic peptides are administered by the parenteral route due to insufficient gastrointestinal absorption. Octreotide, a synthetic human somatostatin analog (SSTa), used for treatment of acromegaly, is not an exception and is parenterally administrated. Recently, a new formulation of oral octreotide was designed based on a Transient Permeability Enhancer ([ldquo]TPE[rdquo]) technology. It consists of octreotide with sodium caprylate in hydrophilic particles, suspended in a lipophilic medium. We characterized the TPE-octreotide intestinal absorption and safety in rats and monkeys.[br][bold]Methods[/bold]: Conscious rats, cannulated at the jejunum and jugular vein, were employed for permeation characterization and pharmacokinetics (PK) studies. Effect on blood GH levels was assessed as well. The intestinal permeation mechanism of TPE was characterized using immunofluorescence and confocal microscopy. Safety was assessed in monkeys after daily oral TPE-octreotide administration for 3-months, and octreotide toxicokinetic analyzed.[br][bold]Results[/bold]: TPE affected tight-junction proteins localization, as demonstrated by following ZO-1 staining pattern, which changed from the characteristically punctuated pattern to an evenly diffused staining through the luminal membrane of enterocytes. Claudin-3 and LC-Biotin, a marker for paracellular penetration, were localized between adjacent enterocyte membranes following TPE administration. The TPE facilitated permeation of dextran molecules up to 20 kD in a transient and reversible fashion. Enteral TPE-octreotide induced absorption was shown to be dose-dependent, with bioavailability reaching up to 8% as compared to s.c. injection. Biotin-labeled octreotide formulated with TPE in rats demonstrated the paracellular absorbance pathway. Octreotide absorbance was unchanged after repeat dosing of TPE-octreotide to rats for 12 days, and a significant suppression of GH levels was detected. Safety assessment after 90-day repeat dosing toxicology study in monkeys showed no test-article related findings. The exposure levels of octreotide were similar across the intervention period, indicating a similar enteral absorption.[br][bold]Conclusions[/bold]: TPE enhanced the intestinal absorption of a biologically active octreotide, plausibly by transient altering tight junction to expand the pores between enterocyts. This formulation was shown to be safe in animals, and is now being clinically studied as an oral alternative for current parenteral SSTa for acromegaly patients.[br][br]Disclosures: ST: Employee, Chiasma, Inc. DP: Employee, Chiasma. AK: Employee, Chiasma. RM: Employee, Chiasma. Nothing to Disclose: ML-A, KM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 397 320 2556 MON-688 PO18-02 Monday 2225 2012


2221 ENDO12L_MON-689 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Clinical Improvement and New Femoral Artery Formation in a Young Man with Peripheral Arterial Disease (PAD) under Human Recombinant Relaxin (hrRLX) and Porcine Relaxin (pRLX) Treatment: An Eight-Year Follow-Up Bernardo Bigazzi, Benedetta Bigazzi, Daniele Bani, Mario Bigazzi Prosperius Institute, Florence, Italy; Prosperius Institute, Florence, Italy; University of Florence, Florence, Italy; Prosperius Institute, Florence, Italy [bold]Background:[/bold] In animal studies, RLX opposed to ischemia and thrombosis, increased vasodilatation and blood flow and caused neoangiogenesis in ischemic tissues (1-2); to date, no applications of RLX have been attempted in human ischemic diseases. Of note, RLX clinical trials in scleroderma (3) and in heart failure (4) reported no negative side effects upon RLX administration.[br][bold]Clinical case:[/bold] In 2003, a 30 year-old man affected by complete obstruction of the superficial femoral and popliteal arteries of the left lower limb (stage IV La Fontaine) came to our observation. Conventional surgical and medical treatments gave no results. A merciful RLX therapy was proposed to prevent amputation.[br]For the first 2 years hrRLX (Connetics), 25 mcg b.wt, was injected s.c., twice a day, for cycles of 3-month therapy and 3-month withdrawal. From the 3rd year to present, oral pRLX, 40 mcg twice a day (Vitalaxin PlusTM,Sky BioHealth, USA) was substituted to hrRLX for better compliance. The general and circulatory conditions have been periodically monitored, clinically and through angiography, angioTC64, photoplethismography and Endo-PAT2000.[br]Left leg circulation showed a dramatic amelioration since the first days of treatment, with higher skin temperature and visual appearance and markedly extended walking range before the onset of claudicatio. Dystrophic toe ulcers and leg hyperpigmentation progressively disappeared. At 3 months, arteriography showed a sharp collateral circulation at the knee level. Leg circulation continuously improved as confirmed by clinical and instrumental follow-up. In 2008, angioTC showed the restoration of blood flow to the left foot due to the appearance of a new elicoidal artery connecting the deep femoral artery to the anterior and posterior tibial arteries.[br]No negative side effects were ever recorded. Circulating anti-RLX antibodies were never found.[br][bold]Conclusion:[/bold] This case report first demonstrates the efficacy of RLX in human peripheral arterial disease. The therapeutic effects were produced not only by vasodilation, but also by anatomical vessel neoformation. HrRLX and pRLX, given s.c., and per os were equally effective, with no negative side effects.[br]This anecdotal case suggests that RLX has the potential to become a powerful therapeutic agent for PAD patients, capable to cause the arrest or even regress of this severe disease.[br][br]1. Bigazzi M, Bani D, Bani Sacchi T. Relaxin: a possible future preventive therapy for cardiovascular disease in postmenopausal women and men? Climacteric 2001, 4: 137-143. 2. Nistri S, Bigazzi M, Bani D. Relaxin as a cardiovascular hormone. Physiology, pathophysiology and therapeutic promises. Cardiovasc Hematol Agents Med Chem (CHA-MC) 2007, 5:101-8. 3. Seibold JR, Korn JH, Simms R, Clements PJ, at all. Recombinant human relaxin in the treatment of scleroderma. A randomized, double-blind, placebo-controlled trial. Ann Intern Med 2000, 132:871-9. 4. Teerlink JR, Metra M, Felker GM, Panikowski P, Unemori E, Teichman SL, at all. Relaxin for the treatment of patients with acute heart failure (Pre-RELAX-AHF): a multicentre, randomised, placebo-controlled, parallel-group, dose-finding phase IIb study. Lancet 2009, 373:1429-39.[br][br]Nothing to Disclose: BB, BB, DB, MB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 398 320 2557 MON-689 PO18-02 Monday 2226 2012


2222 ENDO12L_MON-690 POSTER SESSION: Physiological [amp] Clinical Aspects of GH Secretion (1:30 PM-3:30 PM) Efficacy and Safety of Oral Porcine Relaxin (pRLX) as an Adjunct to Physical Exercise in the Treatment of Peripheral Arterial Disease (PAD) Francesco Sonaglia, Paolo Milia, Bernardo Bigazzi, Benedetta Bigazzi, Marco Caserio, Tito Rastelli, Daniele Bani, Mario Bigazzi Prosperius Tiberino Institute, Umbertide, Italy; Prosperius Institute, Umbertide, Italy; Prosperius Institute, Florence, Italy; Prosperius Institute, Florence, Italy; Prosperius Tiberino Institute, Umbertide, Italy; University of Florence, Florence, Italy; Prosperius Institute, Florence, Italy [bold]Introduction: [/bold]PAD has very poor prognosis and frequently evolves to limb amputation. RLX may be a novel therapy in cardiovascular disease because it induces microvessel dilation, blood flow increase, platelet inhibition and neoangiogenesis, mainly through NO release (1). RLX has not yet been used in vascular ischemic diseases. Aim of our study is to evaluate the efficacy and safety of RLX in PAD.[br][bold]Materials-Methods: [/bold]20 eligible PAD patients (LaFontaine stage 2a_2b), mean age 67,8 M/F 14/2, were scheduled for physical rehabilitation(PR); they were randomized in 1:1ratio in 2 groups. Group A was treated with oral pRLX (Vitalaxin Plus[sup]TM[/sup],Sky BioHealth, USA, 20 mg b.i.d for 12 wk.), group B received placebo and was the control. Clinical monitoring was done weekly. Physical controller training was done on treadmill at 3,2 km/h for 30-60[apos], 3 times weekly. Pain Free Walking Distance (PFWD) and Maximum Walking Distance (MWD) at 3 and 12 wk. were evaluated; follow up 3 months after treatment interruption was performed. Differences were calculated as % variation of individual initial values. Static analysis was done by Student[apos]s test and two-way ANOVA, as appropriate. The study protocol was approved by the Perugia University Ethical Committee.[br][bold]Results: [/bold]Both groups improved during and after the trial. The % increases of PFWD in B group were 23[plusmn]9, 65[plusmn]17, and 35[plusmn]4 respectively at 3 [amp] 12 wk. and 3 months after placebo/PR termination. In comparison, A group showed significantly higher % increases:[br]74[plusmn]16 [italic]p[/italic][lt]0.01, 168[plusmn]28 [italic]p[/italic][lt]0.001, and 122[plusmn]15 [italic]p[/italic][lt]0.001 at the corresponding time points.[br]Similary, the % increases of MWD in the B group were 29[plusmn]7, 55[plusmn]10 and 54[plusmn]8 at the above time points, while in the A group were 55[plusmn]10 [italic]p[/italic][lt]0.001, and 99[plusmn]12 [italic]p[/italic][lt]0.001. The therapeutical action of pRLX appeared very soon and lasted up to 3 months after withdrawal. The RLX patients referred a better physical and mental status. No adverse events during or after the treatment were recorded.[br][bold]Comment: [/bold]RLX resulted very effective in PAD. Our results suggest that the observed functional benefits may come not only from hemodynamic improvement but also from positive vascular remodeling, as suggested by the long-lasting effects, as well as by the evidence of arterial neoformation upon RLX long term therapy in a PAD patient, described by us as a Case Report.[br][br](1) Nistri S, Bigazzi M, Bani D. Relaxin as a cardiovascular hormone. Physiology, pathophysiology and therapeutic promises. Cardiovasc Hematol Agents Med Chem (CHA-MC) 2007; 5; 101:108.[br][br]Nothing to Disclose: FS, PM, BB, BB, MC, TR, DB, MB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 474 320 2558 MON-690 PO18-02 Monday 2227 2012


2223 ENDO12L_MON-691 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Mismatch between Experienced and Objective Stress Markers in Physicians Following 24-Hour On-Call Shifts? Birgit Harbeck, Barbara Pohl, Christian Stefan Haas, Peter Kropp, Hendrik Lehnert, Heiner Moenig University of Luebeck, Luebeck, Germany; Christian-Albrechts-University, Kiel, Germany; University of Rostock, Rostock, Germany Background: Irregular sleep patterns can adversely affect physiological functions and has been associated with increased physiological and psychological stress. Nocturnal working activity of physicians during 24-hour on-call shifts (OCS) disrupts the sleep/wake cycle, thereby causing desynchronization of the natural biological rhythms. The experienced distress has been shown to affect cardiovascular homeostasis and to impair performance of neurocognitive and simulated clinical tasks. However, the clinical impact remains unclear and specific studies are lacking.[br]Objective: To evaluate the effect of a 24-hour on-call shift (OCS) on stress parameters and neurocognitive performance in physicians.[br]Methods: In a prospective cohort study, physiological (e.g. heart rate variability, skin resistance) and biochemical stress parameters were assessed in 11 female and 9 male physicians (median age: 32 years, range: 26 [ndash] 42 years) before and after a 24-hour OCS in Internal Medicine. In addition, various tests of attentional performance (TAP) were conducted.[br]Results: Experienced stress levels during OCS differed from low (n=8) to medium (n=8) and high (n=4). Heart rate variability and skin resistance increased following an OCS, although not statistically significant, while cortisol, adrenaline and noradrenaline levels tended to be lower. Of note, intrinsic alertness was comparable, while phasic alertness was significantly improved following the 24-hour OCS (p[lt]0.05). In addition, vigilance was similar before and after the 24-hour OCS, while focused attention tended to be better following the night shift. These results did not show any correlation with age or medical working experience.[br]Conclusion: We conclude that following a 24-hour OCS: (1) there may be a mismatch between experienced and objective stress levels in physicians; and (2) neurocognitive functions in physicians are not impaired, while performance may even be improved. This may result from long-term gating of attentional properties. Since these data are in contrast to previous reports further studies are needed to study the sequelae of OCS-related stress in physicians and effects on patient care in greater detail.[br][br]Sources of Research Support: Intramural grant of the University of Kiel (B.H.).[br][br]Nothing to Disclose: BH, BP, CSH, PK, HL, HM 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1549 321 2559 MON-691 PO21-02 Monday 2228 2012


2224 ENDO12L_MON-692 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Gonadal and Sexual Dysfunction in Diabetes Patients during Civil Unrest Saad Sakkal Metabolic Care Center, Aleppo, Syrian Arab Republic Objective: Civil unrest is stressful to all. Stress effects metabolic control in general, but no or rare studies showed the influence of civil unrest on gonadal and sexual dysfunction in diabetes patients. We quantitated the effect during the recent Syrian civil unrest (started March 2011).[br]Methods: In our Endocrine/Diabetes clinic patients are seen quarterly. In each visit we review H[amp]P, laboratory parameters and undercurrents psychosocial factors.[br]In 200 patients(96 male,104 female) we determined means FBS,2HPP,HgA1C,Cholesterol, TG, TG/HDL ratio as well as gonadal and sexual functions(libido,ED, gonadal hormones deficiencies) during 2 distinct comparable periods: for the 6 months before the civil unrest and after (July to December 2010, 2011).We excluded new onset diabetes patients and other new illnesses.[br]Results: In the period before civil unrest July to December 2010 There was a drop in Bp (Systolic: 8mm/Diastolic 7mm) FBS (37mg/dl), 2hpp (62mg/dl), HgA1c (2.5%) and insulin dose by 22% from baseline. At the same time there was a drop in Cholesterol (22mg/dl), Triglycerides (59mg/dl), TG/HDL ratio (1.31) implying less inflammation! Gonadal and sexual dysfunction decreased: 17(37 to 20). In the period during civil unrest July to December2011 there was a rise in Bp(26/8)FBS (47), 2hpp (128), HgA1C (2%) and insulin dose by 25%.at the same time there was an increase in Cholesterol(17), TG(115),TG/HDL ratio(1.72)and increase rate of gonadal/sexual dysfunction%100 (16 to32).[br]Conclusion: Diabetes Glucose, lipids, Bp, and gonadal/sexual function are affected by civil unrest, probably as a result of stress, lack of health resources access or other factors. We need more studies for diabetes, Bp, Lipids, and gonadal/sexual status during unfortunate civil unrest events.[br][br]Nothing to Disclose: SS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 153 321 2560 MON-692 PO21-02 Monday 2229 2012


2225 ENDO12L_MON-693 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Moderate Sleep Restriction Modulates Secretory Activity of Somatotropic and Thyreotropic Axis in Healthy Men Sebastian Michael Schmid, Mareike Kueck, Manfred Hallschmid, Kamila Jauch-Chara, Hendrik Lehnert, Bernd Schultes University of Luebeck, Luebeck, Germany; University of Luebeck, Luebeck, Germany; University of Luebeck, Luebeck, Germany; Kantonal Hospital St Gallen, Rorschach, Switzerland Objective: Epidemiological studies point to a strong association between chronic sleep loss and the development of the metabolic syndrome. Clinical studies on sub-chronic sleep deprivation proved distinct alterations of energy homeostasis including decreased insulin sensitivity and energy expenditure, and increased lipolysis. Here, we examined secretory activity of thyreotropic and somatotropic axes following subchronic partial sleep deprivation.[br]Design: In a balanced, within-subject design, circulating concentrations of TSH, fT3, fT4, and growth hormone were closely monitored in 15 healthy, unmedicated normal-weight men during an 15-hours daytime period following two nights of restricted sleep (0245-0700h) and two nights of regular sleep (bedtime 2245-0700h), respectively.[br]Results: Whole day concentrations of fT3 (P[lt]0.026) and fT4 (P[lt]0.089) were elevated, while TSH was progressively suppressed in the evening (P=0.008) after 2 nights of sleep restriction. In contrast to regular sleep, pulsatile growth hormone release during the experimental day was distinctly diminished by sleep restriction (P[lt]0.024).[br]Conclusions: Data indicate a profound impairment in somatotropic axis activity after 2 days of sleep restriction to [sim]4 hours. On the background of elevated fT3/fT4, TSH release was compensatory suppressed after the second night of sleep restriction.[br][br]Sources of Research Support: This work was supported by the Deutsche Forschungsgemeinschaft, SFB 654 [apos]Plasticity [amp] Sleep[apos].[br][br]Nothing to Disclose: SMS, MK, MH, KJ-C, HL, BS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1627 321 2561 MON-693 PO21-02 Monday 2230 2012


2226 ENDO12L_MON-694 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) A Sleep-Related Hypothalamic Signal Modulates Both LH and GH Secretion during Puberty Natalie D Shaw, James P Butler, Janet E Hall Massachusetts General Hospital, Boston, MA; Children[apos]s Hospital Boston, Boston, MA; Brigham and Women[apos]s Hospital, Boston, MA [underline]Context:[/underline] Puberty is marked by a sleep-specific augmentation of GnRH-induced LH secretion[sup](1)[/sup]. The resulting rise in gonadal estradiol drives GH secretion and the pubertal growth spurt[sup](2)[/sup]. We have previously shown that in pubertal children, LH pulses occur in close association with slow-wave sleep (SWS, stage N3)[sup](3)[/sup]. GH secretion is also associated with SWS in children[sup](4)[/sup]. The current study was designed to investigate the relationship of GH and LH pulses to each other and to SWS in pubertal children.[br][underline]Methods[/underline]: Pubertal children with sleep apnea underwent 2 frequent blood sampling studies (q10[apos]x 8hr) with polysomnography. LH and GH pulses were assessed using deconvolution[sup](5)[/sup]. The onset of each pulse was indexed to sleep records to determine the sleep stage at pulse initiation. ANOVA was used to compare: 1) pulse frequency by sleep stage and 2) the % time spent in SWS in 5-min periods before and after each pulse nadir[sup](6)[/sup] to the % SWS across the night. Copulsatility of LH and GH was assessed by visual inspection and by quantifying the co-occurence of LH and GH pulse onsets within a [plusmn] 5 min window. These were analyzed by an extension of traditional copulsatile probabilities[sup](7)[/sup] to include this window of measurement uncertainty.[br][underline]Results:[/underline] 7 subjects, 11-14 yr-old, in Tanner stages II-IV, were studied. During sleep, LH pulses (n=61) occurred more frequently during stage N3 [1.4 [plusmn] 0.2 pulses/hr, mean [plusmn] SE) than during any other sleep stage (p=0.002). The highest frequency of GH pulses (n=64) was during N3 [0.8 [plusmn] 0.1] and N2 [0.8 [plusmn] 0.2] (p=0.002) with no difference between N3 and N2. In all but 1 study, the onset or peak of the initial LH and GH pulse occurred during the first episode of N3. There was significantly more SWS in the 10 min before and after each pulse than expected based on the amount of SWS across the study night for LH (p=0.02), but not for GH (p=0.3). By visual inspection, 25 episodes of copulsatility were observed and in 60% of cases, pulse onset occurred during SWS. Probability analysis indicated 20 episodes of copulsatility, significantly greater than that occurring by chance (p [lt] 10[sup]-6 [/sup]).[br][underline]Conclusion:[/underline] There is significant evidence of copulsatility of GH and LH during sleep in adolescence, particularly with respect to the initial episode of SWS. Thus, in addition to independent hypothalamic stimuli of GH and LH secretion there is evidence of a shared, sleep-related hypothalamic signal which modulates the somatotropic and reproductive axes in puberty.[br][br](1) R Boyar et al. Synchronization of augmented luteinizing hormone secretion with sleep during puberty. NEJM 1972. (2) KC Leung et al. Estrogen Regulation of Growth Hormone Action. Endocrine Reviews 2004. 3() ND Shaw et al. LH Pulses are Associated with Slow-Wave Sleep in Pubertal Children. Endocrine Society Meeting 2011, abstract # 2058. (4) LE Underwood et al. Growth hormone levels during sleep in normal and growth hormone deficient children. Pediatrics 1971. (5) ML Johnson et al. AutoDecon, a deconvolution algorithm for identification and characterization of luteinizing hormone secretory bursts: description and validation using synthetic data. Analytical Biochemistry 2008. (6) JE Hall et al. Brief wake episodes modulate sleep-inhibited luteinizing hormone secretion in the early follicular phase. JCEM 2005. (7) JD Veldhuis et al. Analysis of copulsatility of anterior pituitary hormones. JCEM 1991.[br][br]Sources of Research Support: 5T32HD007396; 1F32HD062315-01A1; Scholars in Clinical Science Program of Harvard Catalyst; Pediatric Endocrine Society; American Medical Association; Endocrine Fellows Foundation; Thrasher Research Fund; Genentech; and the Charles A. King Trust.[br][br]Nothing to Disclose: NDS, JPB, JEH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1782 321 2562 MON-694 PO21-02 Monday 2231 2012


2227 ENDO12L_MON-695 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Effects of Growth Hormone and Testosterone Administration on Thyroid Function in Healthy Older Men Shannon D Sullivan, Ranganath Muniyappa, Jalaja Joseph, S Mitchell Harman, Marc R Blackman Washington Hospital Center, Washington, DC; Georgetown University School of Medicine, Washington, DC; National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), NIH, Bethesda, MD; National Institute on Aging, National Institutes of Health, Baltimore, MD; Kronos Longevity Research Institute, Phoenix, AZ; Phoenix VA Healthcare System, Phoenix, AZ; Johns Hopkins University School of Medicine, Baltimore, MD; Veterans Affairs Medical Center, Washington, DC [bold]Background[/bold]: In childhood and adult growth hormone (GH) deficiency, treatment with exogenous GH decreases circulating thyroid hormone levels and may unmask underlying hypothyroidism. To date, the effects of exogenous GH and/or androgen administration on the hypothalamic-pituitary-thyroid axis in older individuals with low-normal baseline serum concentrations of IGF-I and testosterone (T) have not been reported.[br][bold]Objective[/bold]: To determine the effects of GH and/or T supplementation on thyroid function in healthy older men.[br][bold]Methods[/bold]: A double-masked, 2 x 2 factorial, placebo-controlled, double-dummy design was employed. AM serum concentrations of free T (fT), IGF-I, TSH, total T4 (TT4), and total T3 (TT3) were measured before and after 26 weeks of administration of GH (20 ug/kg sc 3x/week, n=17), T (100 mg T enanthanate IM q2week, n=19), GH+T (n=20) or placebo (n=17) in healthy, older (65-88 yr) men. fT was measured by equilibrium dialysis; IGF-I by RIA; TT4, TT3, and TSH by immunoassay. Mean differences between pre- and post-treatment values ([Delta]) were adjusted for age, baseline value, and treatment group (ANCOVA).[br][bold]Results[/bold]: At baseline, serum concentrations of IGF-I and fT were low to low normal, and those of TT4, TT3 and TSH were normal and similar, in all treatment groups. As expected, GH administration significantly increased serum IGF-I ([Delta]IGF:103[plusmn]80 [micro]g/L, P[lt]0.05), and T administration significantly increased serum fT ([Delta]T: 15[plusmn]22 ng/dL, P[lt]0.05). T administration alone exerted no significant effects on serum TT4, TT3, or TSH. In contrast, GH administration, alone or in combination with T, led to similar decreases in circulating TT4 ([Delta]: GH group, -0.86[plusmn]0.57; GH+T group, -1.1[plusmn]1.3 [micro]g/dL, P[lt]0.05) and reciprocal increases in TSH ([Delta]: GH group, +0.85[plusmn]2.3; GH+T group, +0.87[plusmn]1.4 [micro]IU/mL, P[lt]0.05) without altering TT3.[br][bold]Conclusions[/bold]: These data demonstrate that 6 months of GH administration in healthy older men elicits small but significant decreases in TT4 and reciprocal increases in TSH. In comparison, T supplementation in older men neither directly alters, nor modifies the effects of GH, on serum TT4 or TSH. Whether GH unmasks subclinical hypothyroidism in healthy older men remains to be determined.[br][br]Nothing to Disclose: SDS, RM, JJ, SMH, MRB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 446 321 2563 MON-695 PO21-02 Monday 2232 2012


2228 ENDO12L_MON-696 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Study of the Relation between Serum Melatonin and Immunosenescence Noha Mohamed El Sabbagh, Samir Naim Assaad, Suzan Nashaat Abu Raya, Amina Hesein Hassab, Nany Hassan El-Gayar Faculty of Medicine - University of Alexandria, Alexandria, Egypt; Faculty of Medicine - University of Alexandria, Alexandria, Egypt Aging is associated with a decline in immune function known as immunosenescence. Melatonin release from the pineal gland was observed to have an immunomodulatory role in both animals and humans, and may be altered in the elderly. The aim of this study was to determine the relation of serum melatonin and cortisol to immunosenescence in healthy elderly people.[br][bold][underline]SUBJECTS[/underline][/bold][underline] [bold][amp] METHODS[/bold]:[/underline] The study included 40 healthy individuals (20 men and 20 women) above the age of 65 years, compared to 40 healthy subjects (20 men and 20 women) aged 20-50 years. CD4 and CD8 T lymphocytes were measured in blood by flow cytometry. Serum was used to estimate IgG and IgA (by immunonephelometry), Interleukin 2 (IL-2) (by ELISA), midnight melatonin (by ELISA) and 9 am + midnight cortisol (by ADVIA centaur system).[br][bold][underline]RESULTS[/underline][/bold][underline]:[/underline] Mean serum midnight melatonin was lower in elderly men (34[plusmn]9 pg/mL) and women (35[plusmn]5 pg/mL) in comparison to younger subjects (65[plusmn]13 pg/mL and 68[plusmn]6 pg/mL, respectively)(p=0.0001). Mean serum midnight cortisol levels were higher in elderly subjects versus young individuals. Mean CD8 T cell count was elevated, while CD4 was reduced in elderly men and women in comparison to the younger groups (p=0.001, p=0.001, p=0.001, p=0.01). No significant difference in serum IL-2 and IgA was observed between the elderly and young groups. A significant negative relation was observed between midnight serum melatonin and cortisol in all groups. In elderly men, an inverse correlation was found between CD8 T lymphocyte count and serum melatonin (p=-0.012), as well as between CD4 count and serum cortisol (p=-0.041). No relation was observed between serum melatonin and cortisol, on one hand, and serum IL-2, IgG, or IgA, on the other hand.[br][bold][underline]CONCLUSIONS:[/underline][/bold] In elderly people, there is a reduction in CD4 T lymphocytes and in serum midnight melatonin, and an increase in CD8 T cell count and serum midnight cortisol. An inverse relation exists between CD8 T cell count and midnight melatonin.[br][br]Nothing to Disclose: NMES, SNA, SNAR, AHH, NHE-G 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 222 321 2564 MON-696 PO21-02 Monday 2233 2012


2229 ENDO12L_MON-697 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Development of Impulse Control Disorders in Patients with Dopamine Agonist-Treated Prolactinomas Irina Bancos, Ryan M Nannenga, John M Bostwick, Michael H Silber, Todd B Nippoldt Mayo Clinic, Rochester, MN [bold]Context:[/bold] Dopamine agonists (DA) are widely used in patients with restless leg syndrome, Parkinson[apos]s disease and hyperprolactinemia. In the first two entities, where higher doses of DA are used, an increased frequency of impulse control disorders (ICDs) have been well documented. However, only a few case reports describing ICDs in patients with DA treated prolactinoma have been published.[br][bold]Objectives:[/bold] To determine the frequency of ICDs in patients with prolactinoma treated with DA[br][bold]Design:[/bold] A case-control study: (1) Study group: 100 patients with prolactinoma treated with DA were identified through a search of Mayo Clinic[apos]s medical database, (2) Control group: 327 patients with obstructive sleep apnea (OSA) or restless leg syndrome (RSL) without exposure to DA who served as control group in a similar study at Mayo Clinic, (3) Comparison group: 100 patients with RLS treated with DA (Cornelius JR et al., SLEEP 2010;33(1):81-87). A screening questionnaire for ICDs was used for all three groups. The ICD questionnaire contained modified versions of a compulsive shopping survey, South Oaks Gambling Screen, hypersexuality questionnaire, and punding questionnaires. A positive screen was defined by a preset threshold score for each section.[br][bold]Results:[/bold] With an estimate of 50% response rate, two hundred surveys were sent to patients with diagnosis of prolactinoma and history of DA use. To date, 73 surveys have been returned and are available for analysis. Preliminary analysis of 73 questionnaires for ICDs from prolactinoma subjects showed prevalence of 4.1% for compulsive shopping, 10.4% for pathologic gambling, 11% for hypersexual-ity, and 8.2% for punding. The frequency of any ICD in the study group was 30%, while in the control group [ndash] 15% (OR 2.45, p[lt]0.05). Frequencies of ICDs in the study group were significantly higher for pathologic gambling (OR 9.4, p[lt]0.05) when compared with the control group and there was a trend towards increased development of pathologic shopping (OR 1.7), hypersexuality (2.4) and punding (1.3). When compared to the comparison group frequencies of development of compulsive shopping, gambling, punding and hypersexuality were similar to the study group. There was no relationship to the type of drug used or duration of treatment.[br][bold]Conclusions:[/bold] Preliminary results suggest that ICDs are common in patients with prolactinoma using DA. Counseling before treatment as well as screening for ICDs in this population during treatment should be considered.[br][br]Nothing to Disclose: IB, RMN, JMB, MHS, TBN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1799 321 2565 MON-697 PO21-02 Monday 2234 2012


2230 ENDO12L_MON-698 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Effects of Cabergoline at Zebrafish Development Laura Roesler Nery, Cristiana Hackmann Silveira, Natalia Eltz Silva, Monica Vianna, Mauro Antonio Czepielewski Pontificia Universidade Cat[oacute]lica do Rio Grande do Sul, Porto Alegre, Brazil; Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil Dopamine neurotransmission is a well-established therapeutical target in the treatment of several diseases. Cabergoline (CAB), a D2 agonist has been used for many years in the treatment of Prolactinomas and Parkinson[apos]s Disease. In prolactinoma[apos]s patients, the use of CAB for fertility improvement and during pregnancy remains controversy. Recently CAB was associated with side effects including cardiac disturbances. Zebrafish is a established vertebrate model used for developmental screenings since 1950, and its developmental transparency, easy maintenance, fast growth and efficient absorption of substances in the water, has made him a model for teratogenic and embriotoxicological studies. In this study zebrafish embryos were exposed to CAB (0, 0.1, 1.0 and 10.0 mg/L in system water) for the initial 48hpf. Embryotoxicological evaluation showed no effect on survival and hatching efficiency between groups. The heart rate was diminished in 10.0 mg/L CAB-treated animals at 48hpf but returned to control levels at 10hpf. Apoptotic cell death on heart tissue was evaluated using acridine orange followed by ImageJ quantification of fluorescence at 48hpf and was also increased in 10.0 mg/L-treated animals. At 10dpf, despite the lack persistent effects on heart rate, animals treated with CAB at 10.0mg/L showed altered exploratory capacity, traveling longer distances and making significant more body rotations. All experiments was made in triplicates with 10 embryos/larvae each treatment and statistical analysis was performed with ANOVA followed by Tukey post-hoc (p[lt]0.05). These results demonstrate a deleterious effect of CAB on cardiac development of Zebrafish. A possible comparison with human cardiac disturbance may occur and molecular mechanisms behind those effects need to be investigated.[br][br]Sources of Research Support: Fellowship from Conselho Nacional de Pesquisa (CNPq) awarded to Monica Vianna; Fellowship from Funda[ccedil][atilde]o de Amparo [agrave] Pesquisa do Estado do Rio Grande do Sul (FAPERGS) awarded to Monica Vianna.[br][br]Nothing to Disclose: LRN, CHS, NES, MV, MAC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1142 321 2566 MON-698 PO21-02 Monday 2235 2012


2231 ENDO12L_MON-699 POSTER SESSION: General Neuroendocrinology-Pituitary Axis Function [amp] Dysfunction (1:30 PM-3:30 PM) Etiology of Hypopituitarism in Tertiary Care Institutions in Turkish Population: Analysis of 773 Patients from Pituitary Study Group Database Fatih Tanriverdi, Hatice Sebila Dokmetas, Nur Kebapci, Fatih Kilicli, Hulusi Atmaca, Sema Yarman, Melek Eda Ertorer, Erdinc Erturk, Fahri Bayram, Armagan Tugrul, Cavit Culha, Mehtap Cakir, Meral Mert, Hasan Aydin, Mustafa Taskale, Nese Ersoz, Zeynep Canturk, Inan Anaforoglu, Mesut Ozkaya, Gonca Oruk, Zeliha Hekimsoy, Fahrettin Kelestimur, Tomris Erbas Erciyes University Medical School, Kayseri, Turkey; Cumhuriyet University Medical School, Sivas, Turkey; Department of Endocrinology, Eskisehir, Turkey; Karaelmas University, Zonguldak, Turkey; Istanbul University Faculty of Medicine, Istanbul, Turkey; Baskent University Faculty of Medicine, Adana, Turkey; Uludag University School of Medicine, Bursa, Turkey; Trakya University Medical Faculty, Edirne, Turkey; Ankara Research and Training Hospital, Ankara, Turkey; Selcuk University Meram School of Medicine, Konya, Turkey; Okmeydani Training and Research Hospital, Istanbul, Turkey; Department of Endocrinology, Istanbul, Turkey; Dr Sadi Konuk Research Hospital, Istanbul, Turkey; Hacettepe University Medical School, Ankara, Turkey; Kocaeli University Medical School, Kocaeli, Turkey ; Trabzon Numune Education and Research Hospital, Trabzon, Turkey; Maras Sutcuimam School of Medicine, Kahramanmaras, Turkey; Ataturk Training and Research Hospital, Izmir, Turkey; Celal Bayar University Medical Faculty, Manisa, Turkey [bold]Introduction:[/bold][br]Hypopituitarism in adult life is commonly acquired and main cause is known as pituitary tumors and/or their treatments. However the data regarding etiology of hypopituitarism were obtained from western countries from limited numbers of patients and as far as we know there is only one population-based study in a Spanish population. Since there are new insights regarding the etiology of hypopituitarism and presence of differences in various populations we performed a multi-center database study in Turkish population investigating the etiology of hypopituitarism in 773 patients in Tertiary Care Institutions.[br][bold]Methods:[/bold][br]The study was designed and coordinated by the Pituitary Study Group of SEMT (The Society of Endocrinology and Metabolism of Turkey). Nineteen tertiary reference centers (14 university hospitals and 5 training hospitals) from the different regions of Turkey participated in to the study. The present study is a cross-sectional database study. Data was collected for 18 months starting from August 2007. We classified the causes of hypopituitarism as pituitary adenomas, extra-pituitary tumors and non-tumoral causes.[br][bold]Results:[/bold][br]Mean age of 773 patients (49.8% male, 50.2% female) was 43.9 [plusmn] 16.1 years (range; 16-84 yr). However when we analyze the causes according to gender; the most common etiology in males was pituitary adenomas but the most common etiology in females was non-tumoral causes. According to subgroup analysis of the causes of hypopituitarism in all patients, the most common four causes of hypopituitarism which have frequencies over 10 % were as follows: Non-secretory pituitary adenomas, Sheehan[apos]s syndrome, lactotroph adenomas and idiopathic. With regard to type of hormonal deficiencies; FSH/LH deficiency was the most common hormonal deficit (84.9% of the patients). In 33.8% of the patients 4 anterior pituitary hormone deficiencies (FSH/LH, ACTH, TSH and GH) were present.[br][bold]Conclusions: [/bold][br]Present study clearly demonstrates that Sheehan[apos]s syndrome is still one of the most important causes of hypopituitarism. Further population based prospective studies need to be done to understand the prevalence and incidence of the causes of hypopituitarism in different countries.[br][br]Nothing to Disclose: FT, HSD, NK, FK, HA, SY, MEE, EE, FB, AT, CC, MC, MM, HA, MT , NE, ZC, IA, MO, GO, ZH, FK, TE 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 220 321 2567 MON-699 PO21-02 Monday 2236 2012


2250 ENDO12L_MON-718 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Surgery of Pituitary Tumors in Germany: Hypopituitarism, Mortality, Costs and the Effect of Surgeon Volume Roland Linder, Dietrich Klingmueller, Frank Verheyen TK, Hamburg, Germany; University Hospital, Bonn, Germany The objectives are: to determine the incidence of hypopituitarism and mortality after surgery for pituitary tumors and to ascertain the influence of operation frequencies on the occurrence of these complications.[br]We used a database of 2006-2010 of the Techniker Krankenkasse, one of the biggest statutory health insurance funds in Germany (more than 30 million person years.[br]Between April 1st 2006 and the end of 2010, a total of 480 operations were performed at 102 hospitals. Patients were chosen with ICD-GM E22.0, acromegaly, E22.1, hyperprolactinemia, E24.0, Cushing[apos]s disease and E24.1 Nelson[apos]s syndrome and with the following operating and procedure codes (OPS): 5-075, excision and resection of pituitary gland, 5-010.0, craniotomy, 5-011.2, transsphenoidal approach. Patients with a preoperative need for hormone substitution were excluded. We defined hypopituitarism as the need for hormone substitution according the anatomical therapeutic chemical code (ATC) and/or ICD-GM E23.0, i.e., hypofunction and other disorders of the pituitary gland.[br]Within 3 to 6 months after surgery, no patient died. In this time a hypopituitarism occurred in 29.4% of patients in the two hospitals with the highest operating frequency (mean: 230 operations in 2010). In the remaining 100 clinics (mean: 26 operations in 2010) hypopituitarism occurred in 50.9% of the patients (total: 45.7%).[br]The annual costs of substitution therapy are 1178 [euro] per patient. The costs of lifelong therapy for all patients with new hypopituitarism per year in Germany are calculated to be 22.9 million [euro], assuming a 45.7% rate of hypopituitarism. If the rate of hypopituitarism was only 29.4% about 8 million [euro] would be saved.[br]In conclusion, the database of health insurances is an excellent tool to evaluate therapeutic procedures relatively simple. With these data we confirmed that after surgery for pituitary tumors new hypopituitarism develops in a high percentage of patients. Postoperative hypopituitarism occurred less frequently in higher volume hospitals (often treating even more complicated cases). Results are in line with other papers (1,2). Implications are very important, both for the patient[apos]s quality of life and for economic reasons. To assure quality in surgery, appropriate indicators have to be identified. A minimum quantity regulation might be helpful.[br][br](1)Barker FG; Klibanski A, Swearingen B: Transsphenoidal Surgery for Pituitary Tumors in the United States, 1996[ndash]2000: Mortality, Morbidity, and the Effects of Hospital and Surgeon Volume. J Clin Endocrinol Metab 88(10):4709, 2003. (2)Ciric I, Ragin A, Baumgartner C, Pierce D: Complications of transsphenoidal surgery: results of a national survey, review of the literature, and personal experience. Neurosurgery. 1997;40(2):225-36.[br][br]Nothing to Disclose: RL, DK, FV 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2143 322 2586 MON-718 PO22-03 Monday 2255 2012


2251 ENDO12L_MON-719 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Radiosurgery for Pituitary Adenomas Yamina Suleiman Martos, Ana Roman Cutillas, Ricardo Vilchez Joya, Alfonso Leal Cerro, Jose Maria Martin Linares, Elena Torres Vela Hospital San Cecilio, Granada, Spain; Hospital Virgen de las Nieves, Granada, Spain; Hospital Virgen de las Nieves, Granada, Spain; Hospital Virgen del Roc[iacute]o, Sevilla, Spain [bold]Introduction: [/bold]Sterotactic radiosurgery (SR) has been used to treat recurrent adenomas and also as a primary treatment. The aim of this research is to evaluate long-term tumour control, development of hypopituitarism and other side effects in the follow-up of functioning (FPA) and non-functioning pituitary adenomas (NFA).[br][bold]Patients and method: [/bold]Retrospective analysis of 38 patients with FPA and 21 patients with NFA treated with modified linear accelerator (LINAC). Hormonal control was defined as hormonal normalization. Control of tumour size was defined as an unchange/decrease of tumour size.[br][bold]Results: [/bold]The average follow-up period was 7.3 years for FPA and 6.2 years for NFA. LINAC was the primary treatment in 16 patients with FPA and 3 patients with NFA. There were 4 prolactinomas, 17 adrenocorticotropic hormone-secreting and 17 growth hormone-secreting tumours. Inmunohistochemical data for NFA shows 9 silent pituitary tumours, 4 patients were catalogued as [quot]null cell[quot] and the remaining 8 as NFA in the strict sense. Prior to radiosurgery, pituitary hormone deficits were observed in 23.6% and 66.7% of patients with FPA and NFA respectively. 75%-82.3% of patients with FPA and 52.3% with NFA developed new endocrine deficits during the follow-up. In the group of FPA, 7 patients achieved hormonal remission after SR treatment. The mean dose of treatment was 17 Gy. The decrease in levels of IGF-1 and 24-hour urine cortisol after 3 months of SR was statistically significant (p=0.011 and p= 0.017 respectively), but not in prolactine levels (p=0.144).Tumour volume decreased in 68.4% patients, increased in 5.2% of patients, and was unchanged in 26.3% of patients. In the NFA group the mean dose of treatment was 16.2 Gy. Tumour volume decreased in 52.3% of patients, increased in 38.1% of patients, and was unchanged in 9.5% of patients. Tumour size decrease was statistically significant 12 months after radiosurgery (14.5 mm vs 12.3; p=0.03). Ten patients presented cognitive deterioration after radiosurgery, 2 acromegaly patients and one patient with NFA suffered strokes two- forth years after radiosurgery. One patient developed a second tumour 9 years after SR.[br][bold]Conclusions:[/bold] LINAC resulted in a high and durable rate of tumour in patients with FPA and NFA, hormonal normalization without treatment is more difficult to reach. The follow-up allows the detecting of new endocrine deficits and other side effects that may appear in the long term.[br][br]Nothing to Disclose: YSM, ARC, RVJ, ALC, JMML, ETV 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1287 322 2587 MON-719 PO22-03 Monday 2256 2012


2252 ENDO12L_MON-720 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Pituitary Apoplexy as the First Presentation of a Pituitary Tumor: Retrospective Review of 13 Patients Marta Almeida Ferreira, Ana Rita Caldas, Sergio Moreira, Isabel Ribeiro, Ana Maia Silva, Joana Vilaverde, Isabel Mangas Palma, Andre Couto Carvalho, Fatima Borges Santo Ant[oacute]nio Hospital, Centro Hospitalar do Porto, Porto, Portugal; Santo Ant[oacute]nio Hospital, Centro Hospitalar do Porto, Porto, Portugal [bold]Introduction.[/bold] Pituitary apoplexy (PA) is a life-threatening disorder due to acute ischemic infarction of hemorrhage of a pre-existing sellar mass. It presents with headache, nausea, vomiting, visual impairment, altered mental status and even coma. These non-specific symptoms may delay diagnosis.[br][bold]Aim.[/bold] To review clinical features, management and outcomes of patients with PA admitted to our hospital, from January/2004 to December/2011.[br][bold]Methods.[/bold] Retrospective analysis of medical records was done. Demographical, clinical and histological data was collected and analyzed using descriptive statistics.[br][bold]Results.[/bold] There were 13 patients admitted (9 men; 4 women), with a mean age of 49.8[plusmn]16.1 years-old. The most common presenting symptoms were headache (84.6%, n=11), visual deficits (76.9%, n=10) and nausea/vomiting (61.5%, n=8). All patients were conscious on admission. Eight patients had progressive worsening of visual deficits. No precipitating factors were found. Computorized tomography (CT) and/or magnetic resonance imaging (MRI) of the brain was performed at 0.7[plusmn]0.8 days after admission (range 0-3): all patients had macroadenomas (19.3[plusmn]4.9mm), 84.5% with optic chiasm compression and pituitary stalk deviation. MRI sensitivity for identifying PA was 100% (n=10) [italic]versus[/italic] 77% for CT (n=10). Eleven patients underwent transsphenoidal surgery (4.8[plusmn]4.2 days after admission; range 1-13) while 2 were managed conservatively. Nine patients (75%) presented postoperative hipopituitarism, 6 of whom maintained at least one pituitary hormone deficit. The immunostaining revealed 6 producing tumors: 3 GH; 2 FSH; 1 mixed (prolactin and GH). All symptoms were resolved after treatment with no recurrence (4.2[plusmn]2.3 years of follow-up).[br][bold]Conclusions.[/bold] In this group of patients, PA was more common in men. The most frequent presenting symptom was headache, followed by visual deficits. MRI is the imaging method of choice. All patients had macroadenomas. Transsphenoidal surgery is safe and effective, but hypopituitarism can occur.[br][br]Nothing to Disclose: MAF, ARC, SM, IR, AMS, JV, IMP, ACC, FB 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 100 322 2588 MON-720 PO22-03 Monday 2257 2012


2253 ENDO12L_MON-721 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Are There Imaging Markers of Aggressiveness for Pituitary Tumors? Jean-Francois Bonneville, Francoise Cattin, Nora Clelia Billon-Grand, Veronique Moulin, Alessandra Biondi, Franck Schillo Jean Minjoz University Hospital, Besancon, France; Jean Minjoz University Hospital, Besancon, France From a radiological point of view, we propose to define aggressive pituitary tumors as follows: 1. Tumors with rapid growth as assessed by volumetric measurements by manual contourage or by multiple linear measurements; in any case, precise reproductibility of MR images is requested. This can be easily obtained by using always the same orientation for coronal sections, for instance perpendicularly to the subcallosal plane which remained unchanged with time whatever the evolution of the tumor. 2. Tumors with distinctive MR signal: in GH-secreting pituitary adenomas for instance, invasion of cavernous sinus is more likely to occur with T2 [italic]hyperintense[/italic] tumors, as opposed to [italic]hypointense[/italic] tumors. 3. Tumors extending beyond a natural barrier: capsule, dura, bone. Apart from the historical but not clear-cut features of cavernous sinus invasion, 3.0 Tesla MRI machines have proven here their superiority in demonstrating a rupture or a tear of the very thin dural membrane delineating the interface between cavernous sinus and pituitary fossa. 4. Pituitary adenomas with clival invasion have been recently recognized as aggressive and constitute a potential source of surgical complications. 5. The rare pituitary carcinomas have no specific features but intracranial or systemic metastases. They disseminate via lymphatic and hematogenous spread rather than via craniospinal spread. In conclusion, potential imaging markers of aggressiveness have to be taken into account with clinical, surgical and pathological data when tumoral aggressiveness is suspected.[br][br]Nothing to Disclose: J-FB, FC, NCB-G, VM, AB, FS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 81 322 2589 MON-721 PO22-03 Monday 2258 2012


2254 ENDO12L_MON-722 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Endoscopic Endonasal Approach for Giant Pituitary Adenomas: Advantages and Limitations Maria Koutourousiou, Alessandro Paluzzi, Juan C Fernandez-Miranda, Carl H Snyderman, Sue M Challinor, Paul A Gardner University of Pittsburgh School of Medicine, Pittsburgh, PA; University of Pittsburgh School of Medicine, Pittsburgh, PA; University of Pittsburgh School of Medicine, Pittsburgh, PA [bold]Introduction[/bold]: Giant pituitary adenomas (measuring more than 4cm in maximum diameter) are associated with high rates of residual tumor regardless of the surgical approach. We present the results of the endoscopic endonasal approach (EEA) and analyze the factors that influence the degree of resection.[br][bold]Methods[/bold]: We retrospectively reviewed the medical files and imaging studies of 54 patients (85% male) with giant pituitary adenomas who were managed with EEA.[br][bold]Results[/bold]: The maximum tumor diameter varied from 40 to 90 mm (mean 50mm). Nonfunctioning pituitary adenomas represented 74% of the cases. Preoperative visual impairment was present in 45 patients (83%), partial or complete pituitary deficiency in 28 cases (52%), while 7 patients (13%) presented with apoplexy. Gross total or near total ([gt]90%) tumor resection was obtained in 36 patients (66.7%). Vision was improved or even normalized in 36 cases (80%), remained unchanged in 6 (13%) and deteriorated (due to apoplexy of residual mass) in 2. Pituitary function remained unchanged in 44 patients (81.5%); new pituitary insufficiency occurred in 9 patients (16.7%). Significant factors that limited the degree of resection were a multilobular configuration of the adenoma (p=0.002), extension to the middle fossa (p=0.048) and previous treatment (surgical or medical) (p=0.047). Size, intraventricular extension and invasion of the cavernous sinus did not influence the surgical outcome. Apoplexy at presentation was associated with higher rates of resection. Complications included apoplexy of residual adenoma in 2 cases (3.7%), permanent diabetes insipidus in 5 (9%), transient cranial nerve palsies in 6 (11%), and cerebrospinal fluid leak in 9 (16.7%). After EEA, 13 patients underwent radiotherapy for residual mass and 4 with functional pituitary adenomas received medical treatment. During a mean follow-up of 29.3 months (range 1-109) 7 patients were reoperated for tumor recurrence.[br][bold]Conclusions[/bold]: The main goals of surgery for giant pituitary adenomas are decompression of the optic pathway and maximum safe tumor reduction. EEA provides effective initial treatment of giant pituitary adenomas with fewer limitations compared with the traditional transsphenoidal or open approaches. Despite the satisfactory results with tumor reduction, the high incidence of residual mass frequently requires the use of adjuvant therapies.[br][br]Nothing to Disclose: MK, AP, JCF-M, CHS, SMC, PAG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 994 322 2590 MON-722 PO22-03 Monday 2259 2012


2255 ENDO12L_MON-723 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Short-Stay Protocol Facilitates Early Discharge after Endoscopic Transsphenoidal Resection of Pituitary Adenomas Nisha Gadgil, Susan L Samson, Masayoshi Takashima, Daniel Yoshor Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; St Luke[apos]s Episcopal Hospital, Houston, TX [underline]Background[/underline]: Development of a multi-disciplinary team focused on the care of pituitary patients and the adoption of minimally invasive endoscopic approach, with avoidance of lumbar drains and nasal packing, has enabled shorter inpatient stays after transsphenoidal surgery at our center. However, peri-operative complications including diabetes insipidus, adrenal insufficiency, and CSF leaks remain serious concerns which can delay discharge or result in readmission. [underline]Methods[/underline]: To facilitate safe but early discharge after pituitary surgery, we implemented a protocol that includes evidence-based peri-operative hormonal assessments, education on diabetes insipidus (DI), adrenal insufficiency and CSF leak, with scheduled monitoring after hospital discharge with telephone contact and outpatient clinic visits for the two weeks after surgery. We report the outcomes from 36 consecutive patients who underwent surgery for a pituitary adenoma and were managed with this protocol, with targeted discharge on post-operative day (POD)1. [underline]Results[/underline]: The majority of tumors (81%) were macroadenomas (ave. max. diameter 2.1[plusmn]0.8 cm, range 1.0-4.0) many of which had cavernous invasion (31%), suprasellar (61%), anterior fossa (6%), and/or clival extension (3%). Using the protocol, the median length of stay (LOS) was 1 day (average 1.33[plusmn]0.75 days, range 1-4) with 78% of patients discharged on POD1. Longer stays were attributable to surgical factors in four cases: large and/or recurrent tumors that required extended endoscopic skull base approaches, and one lumber drain. Other factors that contributed to longer LOS included significant medical co-morbidities (intractable hypertension with Cushing[apos]s disease, arrhythmia, multiple sclerosis with impaired respiratory function), and patient anxiety. Diabetes insipidus (DI) occurred in 25% (9 patients) and always presented within the first 24 hours post-operatively. DI did not prolong hospitalization in any patient, and was managed as an outpatient using a protocol that includes daily monitoring of urine output by telephone and dosing of desmopressin under physician guidance. All cases of DI were transient and resolved by POD8 clinic visit. There were no readmissions. [underline]Conclusion[/underline]: Our early experience with a protocl designed to facilitate early discharge of patients after endoscopic pituitary surgery suggests that a majority of patients can be safely discharged less than 24 hours after surgery, with a low risk of complications or readmission.[br][br]Disclosures: SLS: Investigator, Novartis Pharmaceuticals. Nothing to Disclose: NG, MT, DY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1744 322 2591 MON-723 PO22-03 Monday 2260 2012


2256 ENDO12L_MON-724 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Transition from Microscopic to Endoscopic Transsphenoidal Pituitary Surgery: A Single-Center Experience Wibke Billmann, Stefan Bilz, Michael Brandle, Abel-Jan Tasman, Gerhard Hildebrandt, Jean-Yves Fournier Kantonsspital St Gallen, St Gallen, Switzerland; Kantonsspital St Gallen, St Gallen, Switzerland; Kantonsspital St Gallen, St Gallen, Switzerland Background: Due to the potential of improved visualisation and more complete resection endoscopic transsphenoidal pituitary surgery has been increasingly used when compared to the traditional microscopic approach. Various outcome variables before and after the systematic transition from the microscopic to the endoscopic approach at our institution are presented.[br]Methods: The charts of patients who were subsequently referred for transsphenoidal pituitary surgery during the transition from microscopic to endoscopically guided surgery were reviewed. Despcriptive and nonparametric (Mann-Whithney-U-tests) statistics were used for data analysis.[br]Results: Within a 3 years period 31 and 26 patients underwent microscopic (M) and endoscopic (E) pituitary surgery, respectively. Pituitary lesions consisted of endocrine inactive tumors and cysts (17M/18E), GH-producing tumors (11M/3E), ACTH-producing adenomas (3M/2E) and prolactinomas (0M/3E). Percent tumor volume reduction (82 vs. 93%, M vs. E) and the percentage of patients with small ([lt] 1ml) residual tumors (88 vs. 79%) were compareable as were the effects on pituitary function (unchanged 21M vs. 18E, improved 7M vs. 5E, deteriorated 3M vs. 3E). Resolution of hormone excess was achieved in 36 (M) and 38% (E) of the patients. Transient postoperative diabetes insipidus occured more frequently in the endoscopic group (33 vs. 7%), whereas no difference was observed for other perioperative complications (SIAD 3 vs. 7%, cerebral salt wasting 0 vs. 4%, postoperative liquorrhea 13 vs. 15%, postoperative infections 3 vs. 11%, revision surgery 13 vs. 12%, M vs. E).[br]Conclusions: During the transition from microscopic to endoscopic transsphenoidal pituitary surgery a similar extent of tumor resection and cure from hormone excess in endocrine active tumors can be achieved and pituitary function is similarly affected by either approach. A slight increase in perioperative complications, especially transient diabetes insipidus, may occur.[br][br]Nothing to Disclose: WB, SB, MB, A-JT, GH, J-YF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2193 322 2592 MON-724 PO22-03 Monday 2261 2012


2257 ENDO12L_MON-725 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Endoscopic Endonasal Approach for Growth Hormone Secreting Pituitary Adenomas: Outcomes Using 2000 and 2010 Consensus Criteria for Remission Samuel S Shin, Matthew J Tormenti, Alessandro Paluzzi, Sue M Challinor, William E Rothfus, Juan C Fernandez-Miranda, Carl H Snyderman, Paul A Gardner University of Pittsburgh Medical Center, Pittsburgh, PA; University of Pittsburgh Medical Center, Pittsburgh, PA; University of Pittsburgh Medical Center, Pittsburgh, PA; University of Pittsburgh Medical Center, Pittsburgh Background[br]Surgical management of growth hormone (GH) secreting pituitary adenomas has improved over the recent years due to the technical advancements in endoscopic surgery and intraoperative imaging techniques. Most studies reporting outcomes of endoscopic endonasal approach (EEA) on GH secreting pituitary adenomas only used 2000 consensus criteria for defining remission. We compare the remission outcomes using 2010 consensus criteria and 2000 consensus criteria.[br]Methods[br]Retrospective review of 60 patients between 1998 and 2010 at the University of Pittsburgh Medical Center was performed. There were 52 patients who had follow up endocrinological data at least 3 months post injury. Degree of invasiveness was analyzed using Hardy Classification or Knosp Score by an independent radiologist. Patient data was analyzed for complications and disease control using GH levels and IGF-I levels using 2010 and 2000 consensus criteria.[br]Results[br]Using 2010 criteria, remission was achieved in 27 patients (51.9%) receiving EEA only. For patients that didn[apos]t achieve remission by EEA only, additional modalities such as radiosurgery (RS) or medical therapy were combined with EEA. Including the patients treated additionally with other modalities, 37 patients (71.1%) achieved remission. With 2000 criteria, 32 patients (61.5%) achieved remission with EEA only and 44 patients (84.6%) achieved remission with all modalities. Complications included cranial nerve VI palsy (n=1), cranial nerve III palsy (n-1), endocrinological deficits (n=8; 2 developed deficits after RS), cerebrospinal fluid leak (n=4), and meningitis (n=3). Improvements in visual field or acuity from preoperative deficits occurred in 2 patients. No patient had permanent worsening of vision.[br]Conclusions[br]More stringent criteria from 2010 resulted in remission rates approximately 10% lower compared to the 2000 criteria in cases treated by EEA only or in cases with EEA and additional modalities. Despite this lower remission rates, EEA alone or combined with RS and medical management allows high rate of remission while retaining low complication rates.[br][br]Nothing to Disclose: SSS, MJT, AP, SMC, WER, JCF-M, CHS, PAG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 867 322 2593 MON-725 PO22-03 Monday 2262 2012


2258 ENDO12L_MON-726 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Management of Pituitary Non-Functioning Adenomas Amir H Sam, Sachit Shah, Amrish Mehta, Jay Joshi, Karim Meeran, Niamh M Martin, Stephen Robinson, Jeremy Cox, Nigel Mendoza, Federico Roncaroli, Waljit S Dhillo Imperial College London, London, UK Background: The natural history and the optimum management of pituitary NFAs remain unclear.[br]Methods: We carried out a retrospective study of the patients with pituitary NFAs who were either managed conservatively or surgically without postoperative radiotherapy at our centre. Inclusion criteria included: radiological evidence of an adenoma and no evidence of hormone hypersecretion. The pituitary imaging for all patients was re-examined by an experienced neuroradiologist. Patients with a radiological follow-up [lt] 1 year and those presenting with apoplexy were excluded.[br]Results: Sixty-seven patients were managed conservatively for a mean follow-up of 3.8 years (range: 1-14.7). Forty-eight were macroadenomas and 19 were microadenomas. A reduction in the tumour size was observed in 20 (29.9%) patients and the tumour remained stable in 8 (11.9%) patients at the end of the follow-up. In the 39 patients with tumour enlargement, the average time for the first evidence of tumour growth was 1.9 years (0.4-7.5). The average annual growth rate was 1.3 mm/year (0.1- 4.8). One patient developed a visual field defect after 14.7 years. Eight out of seventeen (47.1%) patients with adenomas 5-10 mm in size showed tumour size reduction/no further growth during the follow-up. Twelve out of eighteen (66.7%) patients with adenomas [gt] 10 mm and not impinging on the chiasm had tumour reduction/no further growth during the follow-up. Twenty seven patients with a NFA contacting the optic chiasm did not undergo surgery either due to presence of comorbidities or patient refusal.[br]Forty-eight patients were treated surgically. The NFA contacted the optic chiasm in all patients. The average tumour size was 31.7 mm (13-51.2). The mean post-operative follow-up was 6.2 years (1-19.4). Forty-six patients (95.8%) had residual tumour post-operatively. No tumour enlargement was seen in 22 (45.8%) patients during the follow-up. In the 26 patients with evidence of recurrence, the average time to first radiological sign of tumour enlargement was 3.2 years (0.5-12.2 year). Seven patients (14.6%) had a second operation or further recurrence of optic chiasm compression.[br]Conclusion: Patients with pituitary NFAs without optic involvement can be managed conservatively. Patients with optic involvement must be offered surgery if possible. Post-operatively, patients with residual tumours can be followed up conservatively. Long-term radiological follow-up is necessary in all patients to detect recurrence.[br][br]Nothing to Disclose: AHS, SS, AM, JJ, KM, NMM, SR, JC, NM, FR, WSD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1822 322 2594 MON-726 PO22-03 Monday 2263 2012


2259 ENDO12L_MON-727 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Hyponatremia as a Presenting Sign of Sellar Pathology Huy T Duong, Nancy McLaughlin, Gal Bordo, Amy Eisenberg, Pejman Cohen, Daniel F Kelly John Wayne Cancer Institute, Santa Monica, CA; University of California Los Angles, Los Angeles, CA; University of California Los Angles, Los Angeles, CA Background:[br]Hyponatremia is a relatively rare presenting sign of sellar lesions. Herein we present a series of patients with varying sellar pathology who presented primarily with singular or multiple episodes of hponatremia leading to their diagnosis. [br]Method:[br]Over a 4 year period, all patients who presented with symptomatic hyponatremia as an initial presentation of their sellar mass were included. Lesion pathology, sodium levels, pituitary hormonal status and treatment course were identified.[br]Results:[br]Seven patients (2 males, 5 females, ages 38 to 84) presented with symptomatic hyponatremia with serum sodium levels ranging from 101 to 128 mmol/liter. Of these 7, 3 had two or more episodes of hyponatremia before their sellar lesion was diagnosed. Four patients had pituitary macroadenomas (max diameter 18 to 31 mm; 1 endocrine-inactive vs 3 functional), two patients had arachnoid cysts (max diameter of 27 to 35 mm), and one patient had a Rathke[apos]s cleft cyst (max diameter of 35 mm). Two of 4 pituitary macroadenoma patients also had apoplexy associated with their adenoma. Symptoms included headache, fatigue, and nausea/vomiting and the patient with a 35mm arachnoid cyst also had bitemporal hemianopsia. All patients were admitted and had their hyponatremia corrected with hypertonic saline at our hospital or at an outside hospital. They all underwent uncomplicated endonasal transsphenoidal removal of their pituitary adenoma or drainage of cyst and no patients have had tumor or cyst recurrence. Serum sodium levels normalized in all patients in the early post-operative period and no patients have had recurrent bouts of hyponatremia. One patient had residual headache that resolved at his 3-month postoperative visit. The patient with temporal hemianopsia had complete recovery from her visual field defect. Serum sodium levels range from 132 to 145, 131 to 141, and 137 to 144 mmol/liter on postoperative day 1, 2, and 4 respectively. At 3 months or more after surgery, hormonal status was stable or improved in 6 patients and 1 patient with macroadenoma and apoplexy developed diabetes insipidus, requiring DDAVP.[br]Conclusion:[br]This small series demonstrate that hyponatremia is an important presenting sign of intrasellar pathology. Patients with new onset of hyponatremia and no obvious pharmacologic or other cause should undergo sellar imaging to look for possible pituitary adenoma, Rathke[apos]s cleft cyst, arachnoid cyst, or other intrasellar masses.[br][br]Nothing to Disclose: HTD, NM, GB, AE, PC, DFK 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2336 322 2595 MON-727 PO22-03 Monday 2264 2012


2260 ENDO12L_MON-728 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Association of Clinical Symptoms for Diagnosis of Acromegaly: Identification of Patient Profile. Results of a Pilot Study Charlotte Vaurs, Solange Grunenwald, Jean-Christophe Maiza, Antoine Bennet, Philippe Caron CHU Larrey, Toulouse, France Aim of the study: to identify the association of early symptoms as most characteristic for the diagnosis of acromegaly. Study design: observational, retrospective, transversal, single center, pilot study from patient[apos]s medical records. The identification of the potential association of symptoms was done by analysis of multiple correspondence analysis (MCA). All clinical symptoms were analysed by active variable, except amenorrhea, galactorrhea, dysmorphy and size [lt] 5% (by illustrative variable). Results: data were analysed on 198 patients (46.0[plusmn]14.8 year-old, 62 % females). Most frequent symptoms were extremities infiltration (72%), dysmorphy (68%), headaches (28%), sweating (27%), hypertension (25%), snoring (24%), thyroid goiter (21%), asthenia (17%). The 3 first factorial axes of the MCA do not reflect all information (11.1%, 9.6% and 8.1%). Groups of symptoms which have been identified on those axes were: Axis 1, patients [ge] 60 year-old, Sleep Apnea Syndrome, snoring, diabetes type II, hypertension, thyroid goiter, arthralgia versus female patients [lt] 40 year-old, galactorrhea, amenorrhea. Axis 2, patients with sweating, asthenia, headaches, snoring, arthralgia, versus patients with unilateral carpal tunnel syndrome. Axis 3, men, sexual disorders, visual disorders, Sleep Apnea Syndrome, versus female patients with arthralgia, weight gain, galactorrhea.[br]Conclusion: the results of this pilot study allowed us to identify the association of early or revealing symptoms in acromegalic patients. Due to the number of patients studied, the results need to be confirmed in a larger retrospective and prospective national multicenter study.[br][br]Sources of Research Support: Ipsen Laboratory, France.[br][br]Nothing to Disclose: CV, SG, J-CM, AB, PC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2260 322 2596 MON-728 PO22-03 Monday 2265 2012


2261 ENDO12L_MON-729 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Does Nationwide GH Assay Standardization Affect Post-Surgical Remission Criteria for Acromegaly in Japan? Izumi Fukuda, Naomi Hizuka, Makiko Kurimoto, Yu Yamakado, Toko Muraoka, Kosaku Amano, Yoshikazu Okada, Atsuhiro Ichihara Tokyo Women[apos]s Medical University, Tokyo, Japan; Tokyo Women[apos]s Medical University, Tokyo, Japan [bold]Background: [/bold]In Japan, the growth hormone (GH) assay has been standardized since 2005 through use of a uniform recombinant human GH (rhGH) standard. Since then, GH values measured using the rhGH standard have been approximately 40% lower than previous values measured using kit standards based on the WHO standards for hGH of pituitary origin. However, the Japanese criteria for evaluating treatment outcomes for acromegaly have remained the same: a nadir GH during a 75 g OGTT [lt]1 [micro]g/L is considered cured, 1[le]GH[lt]2.5[micro]g/L is considered inadequately controlled, and [ge]2.5 [micro]g/L is considered poorly controlled, instead of these levels were lowered to 60%, i.e. from 1 to 0.6 [micro]g/L for cured and from 2.5 to 1.5[micro]g/L for inadequately controlled.[br][bold]Objective:[/bold] In this study, we investigated the effects of standardization of the GH assay on the evaluation of post-surgical disease activity in patients with acromegaly.[br][bold]Patients and Methods: [/bold]56 patients with acromegaly (M/F 21/35, 23-72 yr.) were studied. Post-surgical nadir GH during OGTT (within one month) and serum IGF-I levels of the 3 (3M) and 12 months (12M) after TSS were analyzed.[br][bold]Results:[/bold] Post-surgical nadir GH levels during OGTT were positively correlated with the IGF-I SD score 3 months after TSS (Rs=0.46, P[lt]0.05). The normalization rate of IGF-I SDS (12M) in 42 patients whose post-surgical nadir GH[lt]1[micro]g/L (93%, 39/42) did not differ from that in patients whose post-surgical nadir GH[lt]0.6[micro]g/L (94%, 33/35). Among 7 patients whose post-surgical nadir GH ranged between 1 and 2.5mg/L (inadequately controlled), IGF-I SDS (12M) normalized in 4 patients without any additional therapy. Among 8 patients with post-surgical nadir GH [ge]2.5[micro]g/L (poorly controlled), all patients had markedly elevated IGF-I SDS (3M) and required the adjuvant therapy.[br][bold]Conclusion:[/bold] In conclusion, the current Japanese remission criteria for acromegaly still accurately reflect post-surgical disease activity in most patients, as long as one year observation. In this study, IGF-I (12M) normalized without any additional therapy in only 2 of 11 patients whose IGF-I SDS (3M) was elevated, suggesting that persistent elevation of IGF-I might be predicted when the first 3 months IGF-I SDS after TSS is high.[br][br]Nothing to Disclose: IF, NH, MK, YY, TM, KA, YO, AI 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 163 322 2597 MON-729 PO22-03 Monday 2266 2012


2262 ENDO12L_MON-730 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Long-Term Outcome in Acromegaly: Analysis of 1344 Patients from the German Acromegaly Register Christof Schofl, Franz Holger, Martin Grussendorf, Jurgen Honegger, Cornelia Jaursch-Hancke, Jochen Schopohl Friedrich-Alexander-University Erlangen-Nuremberg, Erlangen, Germany; Lohmann and Birkner Health Care Consulting GmbH, Berlin, Germany; Center of Endocrinology and Diabetes, Stuttgart, Germany; Eberhard Karls University Tuebingen, Tuebingen, Germany; German Clinic of Diagnostics, Wiesbaden, Germany; Ludwig-Maximilians-University Munich, Munich, Germany [bold]Introduction:[/bold] Acromegaly is a rare disease with increased morbidity and mortality. Control of growth-hormone excess and normalization of insulin-like growth factor I (IGF-I) reduces morbidity and reverses increased mortality. Currently available therapeutic options have been demonstrated in study populations to allow for disease control in the vast majority of acromegaly patients. Epidemiological data, however, about therapeutic outcome under [ldquo]real life[rdquo] conditions are scarce.[br][bold]Methods:[/bold] 1344 patients (age 55.3 [plusmn] 14.1 yrs, 57 % women) were included from 42 centers participating in the German Acromegaly Register and that had their last follow-up visit within the last 3 yrs. Patients data were collected 10.8 [plusmn] 9.2 (median 8.62) yrs after the initial diagnosis (age at diagnosis 44.8 [plusmn] 13.7 yrs). Controlled disease was defined by an IGF-I within the center-specific reference range. Given are means [plusmn] SD.[br][bold]Results: [/bold]1200 patients (89 %) had at least one surgical intervention, 298 patients (22 %) underwent radiotherapy, and 573 patients (43 %) received medical treatment (somatostatin analogs (n = 407), dopamine agonists (n = 140), pegvisomant (n = 122), combinations (n = 90)). In 917 patients (68 %) acromegaly was controlled according to a normal IGF-1 (158 [plusmn] 58 ng/ml). In patients with an initial diagnosis (ID) dated back [gt] 2 yrs (n = 1012) IGF-1 was normal in 78 % of cases. After surgery 33 % of the patients were controlled without any further therapy. 35 % of the patients with adjunctive radiotherapy had a normal IGF-1 13.75 [plusmn] 9.9 yrs post irradiation. Patients treated medically were controlled in 63 % of cases (77 % ID [gt] 2 yrs). 47 % of the patients with an elevated IGF-1 (432 [plusmn] 228 ng/ml) received no medical therapy.[br][bold]Conclusion: [/bold]Most patients with acromegaly were controlled under real-life conditions in Germany. However, long-term outcome could be improved by exploiting medical treatment options especially in those patients who were not controlled by surgery and/or radiotherapy.[br][br]Sources of Research Support: Novartis Pharma Germany; Ipsen Pharma Germany; Pfizer Pharma Germany.[br][br]Nothing to Disclose: CS, FH, MG, JH, CJ-H, JS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1194 322 2598 MON-730 PO22-03 Monday 2267 2012


2263 ENDO12L_MON-731 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Treatment Modalities and Outcomes for Patients with Acromegaly: A Single-Center Study Jessica MacKenzie-Feder, Lindsey Fisher, Robert Broad, Constance Chik University of Alberta, Edmonton, Canada; University of British Columbia, Vancouver, Canada; University of Alberta, Edmonton, Canada Background: Normalization of insulin-like growth factor-1 (IGF-I) reduces mortality in patients with acromegaly and its associated cardiac, respiratory, rheumatologic, and metabolic complications.[br]Aim: To determine long-term age and sex-adjusted IGF-1 normalization in acromegalic patients in a single centre.[br]Methods: A retrospective review of 131 patients attending a tertiary care centre in Alberta, Canada over the last 30 years was performed.[br]Results: One-hundred and thirty-one patients, 64 women and 67 men (mean age 43.2 (SD13.7)) were followed for an average of 8.3 years (SD 7.9). Sixty-three patients were assessed after 2001. Eight patients were lost to follow up and 8 patients died. Reasons for assessment included acromegalic features, headache, visual field defect, obstructive sleep apnea, colonic polyp, and osteoarthritis. Imaging revealed pituitary macroadenoma in 94 patients (71.8%), microadenoma in 14 (10.7%), normal in 2 (1.5%), and empty sella in 1 (0.8%). Twenty (15.2%) reports were not available. One hundred and seventeen (89.3%) patients had transsphenoidal surgery, 3 (2.3%) had craniotomy, 10 (7.6%) had primary medical therapy (six on somatostatin analogs (SSA), 2 on cabergoline, and 2 on combination), and 1 (0.8%) had primary radiotherapy. Transsphenoidal surgery alone resulted in normalization of IGF-1 levels in 31 (27%) patients. Adjunctive medical therapy included SSA in 70 (53.4%), cabergoline in 12 (9.2%) and pegvisomant in 5 (3.8%) patients. Using multimodal therapy, 94 (72 %) patients achieved sustained normal IGF-1 levels. Four (3.1%) patients refused adjunctive medical treatment. Of 38 (29%) patients who had radiotherapy (37 conventional and 1 stereotactic), only 6 underwent this treatment after 2001. Indications for radiotherapy in these 6 patients included resistance to treatment with SSA or combination therapy in 4 patients, reluctance to be on medication, and financial constraints. Notably, only in the past ten years has octreotide LAR been covered by the provincial health care plan. There is no provincial coverage for pegvisomant.[br]Conclusion: Multiple treatment modalities are required to normalize IGF-1 levels in most patients with acromegaly. Insurance coverage has a major influence on the choice of medical therapy in our patients. The low rate of loss to follow up in our centre is likely attributable to the establishment of a combined pituitary/neurosurgical clinic and a centralized provincial laboratory system.[br][br]Nothing to Disclose: JM-F, LF, RB, CC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 764 322 2599 MON-731 PO22-03 Monday 2268 2012


2264 ENDO12L_MON-732 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Evaluation of Long-Term Effects of Cerebral Radiotherapy on Patients Treated for Acromegaly Iazsmin Bauer Ventura, Juliano Coelho Zakir, Lidia Freire Abdalla, Gabriella Mota, Patricia Barbosa, Amanda Leite Nisiyama, Luiz Augusto Casulari, Luciana Ansaneli Naves University of Bras[iacute]lia, Bras[iacute]lia, Brazil; Laboratorio Sabin, Bras[iacute]lia, Brazil Acromegaly is a multissystemic disorder that requires multiple therapeutic interventions to achieve biochemical control. Conventional radiotherapy may lead to long term disease remission, although high morbidity has been described, as hypopituitarism, cerebro-vascular accidents. Objective: To assess the influence of cerebral radiotherapy in patients previously submitted to surgery and somatostatin analogs. Patients and Methods: This is a cross-sectional study, performed in a neuroendocrine referral center in Brazil. We recruited 27 patients submitted to surgery and radiotherapy with cure criteria of GH[lt]1,0 ng/ml after glucose overload and normal IGF-1 for gender and age (Giustina, 2000), retested them for new criteria GH[lt]0,4 ng/ml, and performed tests to evaluate hiypopituitarism. Results: We enrolled 27 patients, 14 women and 13 men, with mean ages of 38 (20-55) and 29 years(17-51) respectively. All patients were submitted from 1 to 4 surgeries from 12 to 38 months before conventional radiotherapy. The mean time after radiotherapy was 13 years (11-15 years). Most of patients were considered controlled by ancient criteria, but only 18,5% (5/27) of patients achieved GH[lt]0,4 ng/ml and normal IGF-1 levels. From these patients, most had pituitary deficiencies, 48% somatotropic, 44%corticotropic, 46 %gonadotropic, 51% thyrotropic. Conclusion: Biochemical control of growth hormone excess may be improved by complementary radiotherapy, although, the prevalence of hypopituitarism is high, and in our patients, less than 50% achieved the new cure criteria, even after more than 10 years of irradiation.[br][br]Sources of Research Support: Finatec, University of Brasilia, Nucleo de Apoio a Pesquisa Laboratorio Sabin.[br][br]Nothing to Disclose: IBV, JCZ, LFA, GM, PB, ALN, LAC, LAN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 164 322 2600 MON-732 PO22-03 Monday 2269 2012


2265 ENDO12L_MON-733 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Change of Blood Pressure and the Metabolic Markers in before and after Treatment of Patients with Acromegaly Naoki Hiroi, Mariko Sue, Aya Yoshihara, Mayumi Ishikawa, Yasuyo Ando, Gen Yoshino Toho University, Tokyo, Japan [Introduction] It is known that endocrine disorders cause hypertension, and approximately 10% of hypertension is secondary hypertension. The frequency of hypertension caused by acromegaly is the less than 1% in all case of hypertension, however the average life span is shorter in the acromegalic patients than a commoner for 10-15 years, therefore acromegaly should be distinguished at the time of diagnosis of the hypertension.[br][Methods] Fifteen acromegalic patients (7 males, 55.9[plusmn]9.2 years old), who were diagnosed in our hospital from April 2005 to January 2012 were applied, and 6 of 15 cases were patients with hypertension (HT group) and 9 cases were normotension (N group). Before and 6 month after treatment of acromegaly, we obtained measurements of growth hormone (GH) and insulin like growth factor-1 (IGF-1), blood pressure (BP), fasting blood glucose, glycosylated hemoglobin and fasting lipid profiles. All cases were performed transsphenoidal surgery in a neurosurgical institution specialized in pituitary gland, and the postoperative medical treatment was not needed it except in one case.[br][Results] Levels of GH and IGF-1 were decreased from 9.68 [plusmn] 6.65 ng/mL to 1.12 [plusmn] 1.45 ng/mL and from 502.2 [plusmn] 183.48 ng/mL to 181.42 [plusmn] 103.41 ng/mL, respectively. The mean systolic and diastolic BP (SBP and DBP) were 156.0[plusmn]8.7/89.0[plusmn]5.9 mmHg under the use of 1.17 drugs of antihypertensive agents, and were decreased to 124.3 [plusmn] 9.1/73.0 [plusmn] 17.4 mmHg after treatment in HT group (p: [lt]0.005 in SBP). In N group, SBP and DBP were also decreased from 130.7 [plusmn] 3.1/77.3 [plusmn] 2.3 mmHg to 121.5 [plusmn] 2.1/65.5 [plusmn] 19.1 mmHg (p: [lt]0.05 in SBP). There was no significant decrease in the metabolic markers.[br][Discussion] Acromegaly is a chronic progressive disease caused by chronic hypersecretion of GH. It is known that hypertension, diabetes, dyslipidemia are complicated with acromegaly and is reported that hypertension occurs in 18[ndash]41% of this disease. Sodium retention, extraceller fluid volume expansion, and suppression of rennin-angiotensin-aldosterone secretion occur in hypertensive acromegalic patients. Overactivity of the sympathetic nervous system may be involved. In this presentation, hypertension occurs in 40% of acromegalic patients. Before the treatment of acromegaly, the effects of antihypertensive agents were insufficient, however enough depression was observed after the appropriate treatment. It was confirmed that it was appropriate and early diagnosis and treatment were important.[br][br]Nothing to Disclose: NH, MS, AY, MI, YA, GY 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 294 322 2601 MON-733 PO22-03 Monday 2270 2012


2266 ENDO12L_MON-734 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Decreased Physical Activity Level Is Related to Quality of Life and Social Functioning in Patients with Acromegaly Independently of Type of Treatment Renata Elias Dantas, Lara Benigno Porto, Marcia Cristina Reis, Karine Passos, Juliano Coelho Zakir, Barbara Trapp, Luciana Ansaneli Naves Uniceub, Bras[iacute]lia, Brazil; University of Bras[iacute]lia, Bras[iacute]lia, Brazil BACKGROUND: Hypersecretion of growth hormone (GH) due to pituitary adenoma may affect osteo-articular system and cause restriction of physical activity. PURPOSE: The aim of this study was to compare the levels of physical activity with the quality of life in patients with acromegaly, and relate to previous type of treatment for pituitary tumor. METHODS: The sample was composed by 30 patients recruited from the Endocrinology Unit of the University Hospital of Brasilia (HUB). The level of physical activity was evaluated by International Physical Activity Questionnaire (IPAQ 6- short-form), that evaluates the weekly time spent on physical activity of moderate to vigorous intensity in different contexts of life (work, housekeeping, transportation and leisure) and quality of life by The Medical Outcome Study Questionnaire Short Form (SF 36), which is a dimensionless questionnaire to measure generic quality of life, translated and validated for Brazil. This data was compared to activity of the disease and several treatments. Data were analyzed by descriptive statistics. Student[apos]s t test was used, with p [lt] 0.05 ,SPSS 17.0. RESULTS: From all patients, 15 were females, mean age 51,33 [plusmn] 14,33 and 15 were males, mean age 46,2 [plusmn]13,18. The patients were categorized by type of treatment and activity of disease. Artralgia was present in 83% of patients and cephalea was detected in 74%, with no differences in gender. The lowest level of physical activity was related to low functional capability (47,95% IC 37,65-58,25) and emotional aspects (51,52% IC 32,8-70,1). The higher social functioning (75% IC 57,3-92,6) and and general health (75,5% IC 60,49-90,51) were linked to higher physical activity level. CONCLUSIONS: Most of patients presented a reduction in physical activity. This study suggests that there is a relationship between physical activity level and quality of life in acromegalic patients,independently of type of treatment and activity of the disease.[br][br]Sources of Research Support: Finatec, CnPQ.[br][br]Nothing to Disclose: RED, LBP, MCR, KP, JCZ, BT, LAN 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1312 322 2602 MON-734 PO22-03 Monday 2271 2012


2267 ENDO12L_MON-735 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Early Growth Hormone Substitution Following Pituitary Surgery Results in Significant Improvement in Quality of Life and Body Composition Jens Aberle, Nina Sauer, Viola Dannheim, Frank Ulrich Beil, Inga Doeing, Jorg Flitsch Universit[auml]tsklinikum Hamburg-Eppendorf, Hamburg, Germany [bold]Introduction[/bold][br]30-40% of non-functioning pituitary macroadenomas are associated with hyposecretion of the pituitary gland. In addition surgical therapy can lead to a partial or complete hypopituitarism. Data suggest that substitution of growth hormone can improve quality of life and reduce associated symptoms. However in many cases substitution is not started within the first 6-12 months after surgery. Therefore we intended to investigate if patients benefit from an early growth hormone treatment after surgery compared to control.[br][bold]Patients and methods[/bold][br]Data was collected prospectively from 21 adult patients who were diagnosed with insufficiency of growth hormone axis using insulin hypoglycaemia test. Patients were divided into treatment and control group and followed 52 weeks after pituitary surgery. On each consultation laboratory testing was performed and body composition obtained. Also quality of life was assessed by a standardised questionnaire concerning health and life satisfaction.[br][bold]Results[/bold][br]Whereas the control group showed a significant decrease of IGF-1-levels after surgery the IGF1-levels of the treatment group stayed within the normal range.[br]Bioelectrical impedance analysis showed a significant increase of impedance as well as lean body mass in the treatment group compared to control (p[lt]0,05). There was a significant difference in health status in the treatment compared to control.[br]Triglyzerides, cholesterol and LDL-cholesterol increased to a greater extent in the control than in the treatment group without reaching statistical significance. Mean increase in HDL-level was 2.1 mg/dl in the control compared to a decrease of 0.2 mg/dl in the treatment group. Lpa-levels were reduced by 4.1 mg/dl in the treatment and by 2.7mg/dl in the control group. Finally we saw a trend towards better improvement in pituitary function if GH was substituted.[br][bold]Conclusion[/bold][br]Early supplementation of GH leads to a significant improvement in quality of life and body composition after pituitary surgery. It is possible that pituitary function recovers better with early GH substitution.[br][br]Disclosures: JA: Investigator, Novo Nordisk. Nothing to Disclose: NS, VD, FUB, ID, JF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 53 322 2603 MON-735 PO22-03 Monday 2272 2012


2268 ENDO12L_MON-736 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Outcomes of Transsphenoidal Surgery in Patients with Prolactinomas: Emory University Experience Adriana G Ioachimescu, Gena M Mastrogianakis, Samuel Quaynor, Nelson M Oyesiku Emory School of Medicine, Atlanta, GA; Emory School of Medicine, Atlanta, GA Data on surgical outcomes in patients with prolactinomas since introduction of cabergoline is sparse. We retrospectively analyzed all cases of prolactinomas with confirmatory pathology that underwent transsphenoidal surgery by one neurosurgeon between Nov 1999 and Aug 2010. Patients with prior surgery or follow up [lt] 2 months were excluded. Postoperative remission was defined as a normal prolactin level at 2-4 months after surgery without medical treatment.[br]We evaluated 29 patients (15 M, 14 F) ages 33[plusmn]1.8 years (range: 13-58.5) with median follow-up 26.3 months (2-110). Tumor diameter was 1.9[plusmn]1.1 cm (0.5-5). Indications for surgery were inadequate response or intolerance to dopamine agonists, patient[apos]s preference and vision loss. Dopamine agonists were used in 20 patients for 18[plusmn]19 months (range: 1-72) before surgery. Preoperative prolactin in 29 patients in absence of dopamine agonists was 867[plusmn]1009 ng/mL and correlated with tumor diameter (r[sup]2 [/sup]0.74, [italic]p[/italic][lt]0.01). Preoperative prolactin in 20 patients during dopamine agonist treatment was 270[plusmn]363 ng/mL. Postoperative day 1 (POD1) prolactin was 80.9[plusmn]134.1 ng/mL and correlated with the prolactin at 2-4 months postoperatively (50.7[plusmn]66.2 ng/mL, r[sup]2 [/sup]0.62, [italic]p[/italic][lt]0.01), but not with the preoperative prolactin. Prolactin significantly decreased as a result of surgery ([italic]p[/italic] 0.01 versus preoperative prolactin with or without dopamine agonists). Remission was achieved in 9 patients, 5/6 microadenomas and 4/23 macroadenomas. Patients who achieved remission had POD1 prolactin of 1-8 ng/mL and did not recur during follow-up. POD1 prolactin [lt]10 ng/mL predicted remission with 93% sensitivity and 96% specificity. Other predictive factors were tumor diameter (microadenoma), preoperative prolactin and lack of dural invasion. Age and gender did not influence postoperative course, although men had larger tumors ([italic]p[/italic] 0.02) and higher preoperative prolactin ([italic]p[/italic][lt]0.01) than women. Use of dopamine agonists preoperatively did not correlate with biochemical remission. Reoperation was performed in 1 case due to suprasellar residual (craniotomy) and in another due to progression of the tumor residual. Radiotherapy was performed in the same 2 cases due to poor response to dopamine agonists. Transsphenoidal surgery for selected cases of prolactinomas based on current guidelines significantly improves their biochemical control. Our data confirms a strong association of POD1 prolactin with postoperative remission.[br][br]Nothing to Disclose: AGI, GMM, SQ, NMO 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1863 322 2604 MON-736 PO22-03 Monday 2273 2012


2269 ENDO12L_MON-737 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Prolactinomas in Men: A Multicenter and Retrospective Analysis of Treatment Outcome Pedro Iglesias, Carmen Bernal, Carles Villabona, Jose C Castro, Francisco Arrieta, Juan J Diez Hospital Ram[oacute]n y Cajal, Madrid, Spain; Hospital Doce de Octubre, Madrid, Spain; Hospital Bellvitge, Barcelona, Spain; Hospital General, Segovia, Spain Aims: To assess treatment outcome in male patients with micro- and macroprolactinomas.[br]Design: Multicenter, retrospective study.[br]Patients: 88 male patients (15 micro- and 73 macroprolactinomas), aged 40.3 [plusmn] 14.7 years were studied. Time of follow-up ranged from 3 to 244 months.[br]Methods: Clinical, hormonal and radiological data were registered at diagnosis and follow-up. Treatment outcome was evaluated in relation to the modality of therapy (dopamine agonists, surgery and radiation therapy).[br]Results: Dopamine agonists normalized prolactin levels in 73.3% and 65.2% of patients with micro- and macroprolactinomas, respectively. Disappearance of tumor was reached in 53.3% and 28.3% of subjects with micro- and macroprolactinomas, respectively. Tumor absence at last visit was achieved in 7 out of 14 patients with macroprolactinoma and treated by means of dual therapy (dopamine agonists and neurosurgery) and in 9 out of 13 patients with macroprolactinoma managed with triple therapy (dopamine agonists, neurosurgery and radiation therapy). Normalization of prolactin levels at last visit was present in 68.9%, 79.6% and 69.2% of patients treated by medical therapy, dual therapy and triple therapy, respectively (differences not significant). Multivariate logistic regression analysis showed that the time on therapy was the only significant variable related to tumor disappearance.[br]Conclusion: We conclude that medical therapy normalizes prolactin and reduces tumor size in the majority of men with prolactinomas. The addition of pituitary surgery with or without radiation therapy does not offer significant advantages over medical therapy with dopamine agonists in male patients with macroprolactinomas.[br][br]Nothing to Disclose: PI, CB, CV, JCC, FA, JJD 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 47 322 2605 MON-737 PO22-03 Monday 2274 2012


2270 ENDO12L_MON-738 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Hyperprolactinemia Due to Chronic Nipple Piercing [mdash] An Urban Legend? Grace Sun, Kevin Pantalone, Manjula Gupta, Marigel Constantiner, Amir Hamrahian, Laurence Kennedy, Christina Reed, Betul Hatipoglu Cleveland Clinic Foundation, Cleveland, OH; Summa Western Reserve Hospital Physicians, Inc, Hudson, OH; Cleveland Clinic Foundation, Cleveland, OH; Cleveland Clinic Foundation, Cleveland, OH; Cleveland Clinic Foundation, Cleveland, OH [bold]Background[/bold]: The literature on elevated serum prolactin levels in patients with nipple piercing is limited to those with concomitant breast/chest wall infection. It is unclear if chronic nipple stimulation from a piercing alone can cause sustained elevations of serum prolactin. The aim of this study was to determine if either men or women with nipple piercing longer than 6 months duration have elevations in serum prolactin.[br][bold]Methods[/bold]: This cross-sectional cohort study was approved by the institutional review board. Inclusion criteria were as follows: men and women [ge]18 years old with nipple piercing(s) present [gt]6 months. Exclusion criteria included: women who are pregnant, lactating or [lt]6 months postpartum; subjects on medications known to increase prolactin levels; chest wall/breast infection at the time of blood draw or conditions known to be associated with hyperprolactinemia. All samples were analyzed by the Cleveland Clinic Reference Laboratory. Normal serum prolactin values are 2-17.4 ng/mL in non-pregnant women and 2-14 ng/mL in men.[br][bold]Results[/bold]: A total of 3 men and 5 women were enrolled. The median (range) ages for men and women were 33 years (24-42) and 30 years (23-34), respectively. Seven of 8 were Caucasian; one female was Filipino. All subjects had bilateral piercings. The median interval from nipple piercing to blood draw was 3 years (2-12). None of the subjects had hyperprolactinemia. Median prolactin levels for men and women were 5.6 ng/mL (3.8-7.4) and 8.3 ng/mL (4.6-10.2), respectively.[br][bold]Conclusion[/bold]: Our results suggest that in the absence of any concomitant skin infection, chronic nipple piercing is not associated with hyperprolactinemia. Although the extrapolation of results is limited given the small sample size of relatively young and healthy adults, our findings suggest that hyperprolactinemia in a patient with a chronic nipple piercing warrants further investigation.[br][br]Nothing to Disclose: GS, KP, MG, MC, AH, LK, CR, BH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 259 322 2606 MON-738 PO22-03 Monday 2275 2012


2271 ENDO12L_MON-739 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) The Diagnostic Usefulness of Thyrotropin-Releasing Hormone (TRH) and Metoclopramide Stimulation Tests in the Evaluation of Hyperprolactinemia Deepa Beeharry, Gayatri Sreemantula, Jiten Vora, Dushyant Sharma Royal Liverpool University Hospital, Liverpool, UK Aims:[br]The objective of our study was to investigate the usefulness of the dynamic pituitary tests in the evaluation of hyperprolactinaemia.[br]Methods:[br]A retrospective audit was performed in 104 patients (31 male and 73 female; age range from 26 to 83 years) with hyperprolactinaemia who were evaluated by a metoclopramide and/or TRH stimulation test at the Royal Liverpool University Hospital, Liverpool, UK between 2006 and 2009. Macroprolactinaemia, renal failure and hypothyroidism were excluded in these patients. The prolactin response to 200 mcg of intravenous synthetic TRH and 10 mg of intravenous metoclopramide was measured under standardised conditions. Serum prolactin level was checked at baseline, 30 minutes and 60 minutes. A less than 2.5 fold rise from baseline prolactin is expected in patients with microprolactinoma ([lt]10 mm) and macroprolactinoma ([gt]10 mm) while a greater than 2.5 fold rise is expected in patients with normal imaging.[br]Results[br]Radiological imaging (N = 84) showed structural lesions in 38 patients: microprolactinoma (N =19), macroprolactinoma (N = 7), other radiological diagnosis e.g craniopharyngioma, null cell adenoma (N = 12). All patients with macroprolactinoma and 46% (7/15) of patients with microprolactinoma showed [lt] 2.5 fold rise in prolactin level following TRH and metoclopramide stimulation. Amongst people with normal imaging, 58% (14/24) and 29% (13/45) showed an expected ([gt] 2.5 fold rise) in prolactin levels in the metoclopramide and TRH tests respectively. In the category of [apos]other radiological diagnosis[apos], 80% (8/10) showed a less than 2.5 fold rise in prolactin.[br]Conclusion:[br]Our study supports that patients with a radiological diagnosis of macroprolactinoma show [lt]2.5 fold rise in prolactin following TRH and Metoclopramide stimulation. However, variable degree of prolactin response suggests that the role of these dynamic pituitary tests is uncertain in the differentiation of microprolactinoma, other pituitary lesions or idiopathic (normal imaging) hyperprolactinaemia. A larger randomised controlled trial is required to establish the role of these dynamic pituitary function tests, where the radiological picture alone does not explain the underlying aetiology of hyperprolactinaemia.[br][br]Nothing to Disclose: DB, GS, JV, DS 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1607 322 2607 MON-739 PO22-03 Monday 2276 2012


2272 ENDO12L_MON-740 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Profile of Patients with Hyperprolactinemia Accompanied by a Tertiary Hospital in the Northeast Brazil Jose Italo Mota, Wladia Gomes de Paula Sobreira, Francisco Assis Alves Teixeira Junior General Hospital of Fortaleza, Fortaleza, Brazil Hyperprolactinemia is a major hormonal change of the hypothalamic-pituitary axis. This study aimed to draw a clinical and epidemiological profile, including treatment, of patients with hyperprolactinemia followed by the General Hospital of Fortaleza (HGF). It consists in a transversal, analytical, descriptive, retrospective and quantitative study, conducted by patient records review. The study population comprised 83 patients with hyperprolactinemia assisted by the Endocrinology Service of HGF. The sample used is the non-probabilistic intentional, where the selection included patients with hyperprolactinemia with last appointment between March and September 2010. 63 (81.9%) patients were female and 15 (18.1%), male. The highest percentage of patients (48.2% (40/83) had microprolactinomas, followed by 43.4% (36/83) with macroprolactinomas. The most frequent symptom was galactorrhea, followed by amenorrhea, menstrual irregularity and headache. The majority of micro and macroprolactinomas were treated with dopamine agonists. 61.5% (16/26) of microprolactinomas and 63%(17/27) of macroprolactinomas reduced by 50% or greater tumor volume or had no evidence of tumor after treatment (p[lt]1,000). 30/33 (90.9%) and 12/19 (63.2%) of micro and macroprolactinomas respectively obtained the regularization of the menstrual cycle (p[lt]0,026). 33/33 (100%) of micro and 15/17 (88.2%) of the macroprolactinomas improved galactorrhea (p[lt]0,126). 9/15 patients with macroprolactinomas reported improvement of visual symptoms. In this study, prolactinoma was the major cause of hyperprolactinemia; dopamine agonists are widely accepted as a primary therapy, with well tolerance and effectiveness. Close and long-term follow-up is needed to achieve improvement in the resolution of the signals and symptoms, since the treatment can be extended for many years.[br][br]Nothing to Disclose: JIM, WGS, FAATJ 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2379 322 2608 MON-740 PO22-03 Monday 2277 2012


2273 ENDO12L_MON-741 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Characterization of Risk and Long-Term Outcomes in Treated Cushing Disease Jessica Kehris Lambert, Levana Goldberg, Sofia Fayngold, Jane Kostadinov, Kalmon D Post, Eliza B Geer Mount Sinai School of Medicine, New York, NY; Mount Sinai School of Medicine, New York, NY; Mount Sinai School of Medicine, New York, NY [bold]Background[/bold]: Cushing[apos]s Disease (CD) confers a fourfold increase in mortality and is associated with significant morbidities (1). Cardiovascular (CV) risk may persist up to five years after endocrine cure (2, 3) and studies looking more than 10 years post-operatively demonstrate that treated CD is associated with excess mortality (4-6). Characterization of predictors of increased morbidity and mortality among patients with treated CD requires further elucidation. [bold]Objective:[/bold] This study aims to identify predictors of cardiovascular disease, tumor recurrence, and mortality after long-term follow up among patients with treated CD. [bold]Methods: [/bold]A retrospective chart review was conducted evaluating 334 patients with a confirmed diagnosis of CD who underwent transphenoidal adenectomy with a single surgeon. A database was created, including: age at diagnosis; length of exposure to excess glucocorticoids (estimated based on duration of symptoms prior to diagnosis); co-morbidities at diagnosis; MRI, operative, and pathologic data; treatment modalities; pre and post-operative laboratory data. Cox Proportional Hazard Models identified predictors of mortality, recurrence and adverse CV outcomes. [bold]Results: [/bold]323 subjects were included. Mean age was 39.5 +/- 14.2 years; mean follow-up was 81.4 +/- 75.5 months with an estimated duration of exposure to excess glucocorticoids of 36.6 +/- 35.2 months, including pre and post-operative exposure. Multivariate analyses demonstrated that increased duration of symptoms prior to diagnosis elevated the risk of death (HR, 1.016 [95% confidence interval {CI}, 1.006-1.025], p [lt]0.002). Age at diagnosis elevated the hazard both of death (HR, 1.053 [CI, 1.019-1.088], p[lt]0.001) and CV events (HR, 1.066 [CI, 1.026-1.107], p[lt]0.001). Preoperative ACTH levels increased the hazard of death (HR, 1.013 [CI, 1.006-1.020], p[lt]0.001). Male sex increased the hazard for cardiovascular events (HR, 2.645 [CI, 1.006-6.970], p[lt]0.049). A normal preoperative pituitary MRI increased the risk of cardiovascular events (HR, 4.233 [CI, 1.640-10.925], p[lt]0.003). [bold]Conclusion:[/bold] Long-term follow up of a large cohort of treated CD patients demonstrates that older age at diagnosis, higher pre-operative ACTH, and longer exposure to excess glucocorticoids are associated with decreased survival. Our data emphasize the importance of early diagnosis and treatment of CD. Improved understanding of the morbidities associated with treated CD is needed to provide appropriate interventions.[br][br](1) Pivonello et al. The metabolic syndrome and cardiovascular risk in Cushing[apos]s syndrome. Endocrinol Metab Clin North Am 2005; 34: 327-339. (2) Extabe J, Vazquez JA. Morbidity and mortality in Cushing[apos]s disease: an epidemiological approach. Clin Endocrinol (Oxf). 1994; 40: 479-484. (3) Colao A et al. Persistence of increased cardiovascular risk in patients with Cushing[apos]s disease after five years of successful cure. JCEM. 1999. 84; 2664-2672. (4) Hammer GD et al. Transphenoidal Microsurgery for Cushing[apos]s Disease: Initial Outcome and Long-Term Results. JCEM. 2004; 89: 6348-6357. (5) Dekkers OM et al. Mortality in Patients Treated for Cushing[apos]s Disease Is Increased, Compared with Patients Treated for Nonfunctioning Pituitary Macroadenoma. JCEM. 2007; 92: 976-981. (6) Clayton RN et al. Mortality and Morbidity in Cushing[apos]s Disease over 50 Years in Stoke-on-Trent, UK: Audit and Meta-Analysis of Literature. JCEM. 2011; 96: 632-642.[br][br]Nothing to Disclose: JKL, LG, SF, JK, KDP, EBG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 579 322 2609 MON-741 PO22-03 Monday 2278 2012


2274 ENDO12L_MON-742 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Long-Term Cure and Recurrence Rates in Cushing Disease: Predictive Factors in a Single-Center Study Krystallenia I Alexandraki, Gregory A Kaltsas, Andrea M Isidori, Helen Storr, Farhad Afshar, Ian Sabin, Scott A Akker, Shern L Chew, William M Drake, John P Monson, G Michael Besser, Ashley B Grossman St Bartholomew[apos]s Hospital, Barts and the London NHS Trust, and Barts and the London School of Medicine, London, UK; St Bartholomew[apos]s Hospital, Barts and the London NHS Trust, and Barts and the London School of Medicine, London, UK; St Bartholomew[apos]s Hospital, Barts and the London NHS Trust, and Barts and the London School of Medicine, London, UK Context: Cushing[apos]s disease (CD) is the most common cause of endogenous hypercortisolemia with transsphenoidal adenomectomy the preferred initial treatment.[br]Objective: To investigate the early and late outcomes of patients with CD submitted to a neurosurgical procedure as first-line treatment.[br]Design: Single-center retrospective case-notes study.[br]Setting: University hospital.[br]Patients: 131 patients with CD with a minimum follow-up period of 6 years.[br]Main Outcome Measure(s): Cure: post-operative 09.00h serum cortisol level [lt]50nmol/L; remission: cortisol insufficiency or restoration of [apos]normal[apos] cortisol levels with resolution of clinical features; recurrence: dexamethasone-resistance and relapse of hypercortisolemic features.[br]Results: Seven patients had transcranial and 124 transsphenoidal approaches. Of this latter group 67.7% obtained remission and 45.3% cure with recurrence rates of 23.8% and 15.1% respectively (15.0 [plusmn] 6.2years of follow-up). Of 23 patients with microadenomas operated after 1991 and with positive imaging and pathology, 91.3% obtained remission and 69.6% cure; recurrence was found in 9.3% and 6.2% respectively. In multivariate analysis, the time needed to achieve hypothalamic-pituitary-adrenal axis recovery was the best predictor of recurrence; all patients who recurred showed recovery within 3 years from surgery. No recurrence was seen in the group of patients who had a total hypophysectomy. Strict cure criteria did not prove to be superior in terms of the probability of recurrence compared to postoperative normocortisolemia.[br]Conclusions: Life-long follow-up for patients with CD appears essential, particularly for patients who have shown rapid recovery of their axis. The strict criteria previously used for [apos]apparent cure[apos] do not appear to necessarily predict a lower recurrence rate.[br][br]Sources of Research Support: Dr K. Alexandraki was awarded a scholarship by the Alexander S. Onassis Public Benefit Foundation.[br][br]Nothing to Disclose: KIA, GAK, AMI, HS, FA, IS, SAA, SLC, WMD, JPM, GMB, ABG 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 634 322 2610 MON-742 PO22-03 Monday 2279 2012


2275 ENDO12L_MON-743 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Endoscopic Endonasal Transsphenoidal Surgery: Results in 28 Consecutive Patients with Cushing Disease Jason M Ng, Alessandro Paluzzi, Juan C Fernandez-Miranda, Paul Gardner, Sue M Challinor University of Pittsburgh, Pittsburgh, PA; University of Pittsburgh, Pittsburgh, PA [bold]Purpose: [/bold]The conventional transsphenoidal approach to pituitary surgery for Cushing[apos]s Disease (CD) achieves remission in [sim]80% (1). Surgical complications include a 2-41% (ave.20%) risk of hypopitutarism, 3-9% risk of diabetes insipidus (DI), and 0-8% risk of CSF leak (2). The endoscopic endonasal approach (EEA) was developed as an alternative, utilizing a nasal passageway to permit a less invasive procedure. In a recent report, this approach achieved a remission rate of 83% in 35 patients with CD and caused hypopituitarism, excluding hypoadrenalism, in 48% (3). The aim of our study is to report the results of EEA in a consecutive series of patients with CD and determine remission rates, and surgical complications of this procedure.[br][bold]Methods: [/bold]We retrospectively reviewed patients with CD who underwent EEA and were diagnosed and treated, with available follow-up data, at our institution from 10/2005 [ndash] 11/2009. Remission was defined as a postoperative morning cortisol of [le]5 mcg/dl within 1 week of surgery and at 6 months post-op, a low to normal 24 hour urine free cortisol, and no symptoms of CD (1,4). In total, 28 patients were assessed, 5 were excluded because their diagnostic workup for CD did not meet established criteria (5,6).[br][bold]Results: [/bold]Mean age was 47.6 years (range 31-69 years) with mean follow-up of 15.6 months (range [frac12]-43 months) and tumor size range of 2-14 mm. Remission was achieved with the first surgery in 19 of 23 patients and 4 patients required a second surgery. New anterior pituitary hormone deficits, excluding hypoadrenalism, occurred in 6 (26%). Two patients (8%) had permanent Diabetes Insipidus (DI). Deficits included hypothyroxinemia and adrenal insufficiency (AI) in 2 patients, persistent AI alone in 6 patients, growth hormone (GH) deficiency in 1 patient, and GH, thyroid, gonadotropin, and AI in 3 patients. In these 3 patients, 1 had permanent DI and 2 had transient DI. An additional 2 patients had transient DI and 1 patient had permanent DI, only. Eleven patients required glucocorticoid therapy at last follow up, with mean follow-up of 16.9 months (range 5-43 months). There were no post-operative CSF leaks, vision loss, or cranial nerve defects. One patient had anosmia.[br][bold]Conclusion: [/bold]The EEA for ACTH secreting pituitary adenoma resection provided an excellent remission rate of 82 % in our series and can be considered a safe alternative to the traditional microscopic approach with few complications in patients with CD.[br][br]1. Hammer GD et al., J Clin Endocrinol Metab 2004; 89:6348. 2. Kelly DF, J Neurosurg. Focus 2007; 23:E5. 3. Netea-Maier RT et al., Eur J Endocrinology 2006; 154:675. 4. Esposito F et al., J Clin Endocrinol Metab 2006; 91:7. 5. Arnaldi G et al., J Clin Endocrinol Metab 2003; 88:5593. 6. Nieman LK et al., J Clin Endocrinol Metab 2008; 98:1526.[br][br]Nothing to Disclose: JMN, AP, JCF-M, PG, SMC 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2277 322 2611 MON-743 PO22-03 Monday 2280 2012


2276 ENDO12L_MON-744 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Postoperative Cortisol and ACTH in Patients with Pathologically Confirmed ACTH-Secreting Pituitary Adenoma: Role in Predicting Remission of Cushing Disease Nadia Hameed, Chris Yedinak, Jessica Brzana, Nicholas Coppa, Johnny Delashaw, Sakir Gultekin, Aclan Dogan, Maria Fleseriu Oregon Health [amp] Science University, Portland, OR; Oregon Health [amp] Science University, Portland, OR; Oregon Health [amp] Science University, Portland, OR Introduction: Postoperative serum cortisol (C) is used as an indicator of Cushing[apos]s disease (CD) remission following transsphenoidal surgery (TSS) and guides (controversially) the need for immediate adjuvant treatment for CD or hydrocortisone (HC) replacement.[br]Methods: We investigated postop C and ACTH as predictors of remission/recurrence in CD in a retrospective cohort of 54 pts with pathologically confirmed CD (2006-2011) at OHSU. Midnight and morning C, ACTH at 24-48 h postop ([gt] 24 h after last HC dose) were measured in 40 pts. Remission, defined as nl 24-h UFC, nl midnight salivary C, nl Dexamethasone CRH test or continued need for HC, was assessed periodically. Statistical analysis was performed using PAWS 18.[br]Results: Follow up data was available for 52 pts; median follow up 16.5 mo (2-143); F/M 38/14, median age 45 y (21-72 y); 28 micros(m), 16 macros, 8 no tumor on MRI. 9/52 pts with C [gt]10 [micro]g/dl by postop day 1-2 had 2nd TSS within 7 days.[br]43 pts (82.7%) achieved CD remission (37 after 1 TSS, 6 after a 2nd early TSS), 6 pts suffered disease recurrence (mean 39.2 [plusmn] 52.4 mo). An immediate 2nd TSS induced additional hormonal deficiencies (DI) in 2 pts; no surgical complications. Persistent disease was noted in 9 pts despite 3 pts having an immediate 2nd TSS.[br]Positive predictive value for remission of C [lt]2 [micro]g/dl and ACTH [lt]5 pg/ml was 100%. C, ACTH levels (at all time points postop and at 2 mo were correlated (r=0.37, p[lt]0.001).[br]Nadir serum C of [le]2 [mu]g/dl and ACTH [lt]5 pg/ml predicted remission (p[lt] 0.005), but no level predicted lack of recurrence. Immediate postop ACTH/C did not predict length of remission.[br]No pts with postop C [gt]10 [mu]g/dl were observed to have delayed remission; all required additional treatment.[br]There was no significant difference in age, BMI, tumor size and length of follow-up between postop C groups: C[le]2 [mu]g/dl, C [gt]5 [mu]g/dl and C [gt]10 [mu]g/dl.[br]Conclusions: Immediate postop C levels should routinely be obtained in CD pts post TSS, until better tools to identify early remission are available. Immediate repeat TSS could be beneficial in pts with C [gt]10 [mu]g/dl and positive CD pathology: our combined (m and macros) remission[apos]s rate with this approach was 82.7%. ACTH measurements correlate well with C. However, because no single C or ACTH cutoff value excludes all recurrences, pts require long-term clinical and biochemical follow-up. Further research is needed in this area.[br][br]Nothing to Disclose: NH, CY, JB, NC, JD, SG, AD, MF 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1749 322 2612 MON-744 PO22-03 Monday 2281 2012


2277 ENDO12L_MON-745 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) ACTH-Staining Adenoma Removal and Changes in Weight, Blood Pressure and Blood Glucose Level Sann Yu Mon, Laurence Kennedy, Betul A Hatipoglu Cleveland Clinic, Cleveland, OH OBJECTIVE: Screening for Cushing[apos]s disease is not routinely performed before incidental macroadenoma removal unless clinically indicated. We decided to compare pre and post-op body weight, blood pressure and blood glucose in ACTH staining pituitary macroadenomas. SUBJECT AND METHODS: Subjects were identified from a pituitary database at the Cleveland Clinic. We retrospectively reviewed 92 patients with nonfunctional pituitary macroadenoma who underwent transspenoidal surgery. Pre and post-op blood pressure, weight in kg and random blood glucose in ACTH staining group were compared, none of them had any signs for Cushing[apos]s. RESULTS: Among 92 specimens, 5 stained strongly, 7 weakly for ACTH. All 5 patients with strong staining had weight reduction. Pre-op weights were 62.28, 58.97, 72.44, 108, 107 kg and post-op 6-12 months became 57.56, 52.66, 65, 98.39, 105.24 kg respectively. 3 of them had better blood pressure control with no medication change. Pre-op BP were 132/92, 147/92, 140/80, 150/90, 144/98 mmHg and post op were 133/87, 158/86, 111/64, 112/82 and 124/78 mmHg respectively. No changes in blood glucose were noted. Pre-op basal cortisol levels were15.3, 23.1, 2.0, 9.2, 9.7 mcg/dL. 4 patients had elevated pre-op ACTH levels of 42.0, 20.0, 81.0, 109.0 pg/ml. Post-op cortisol levels were 11.7, 4.0, 45.0, 12.6, 10.3 mcg/dL respectively. 2 were started on levothyroxine ,one on hydrocortisone. Among the 7 patients with weak staining, 3 patients had weight reduction, 1 patient had weight gain and 2 improved blood pressure control. For 7 patients pre vs post op were 134.27 vs. unknown; 88.91 vs 88.45; 78.16 vs. 80.51; 96.5 vs 96.1; 97.75 vs 96; 126.1 vs 124.79, 96.61 vs 94.35 kg respectively. Pre op vs. post op BP for those 7 patients were 147/74 vs 122/90; 138/80 vs 120/60; 138/88 vs 147/95; 120/69 vs 118/81; 144/86 vs 138/80; 134/85 vs 128/76; 130/70 vs 122/60 mmHg respectively. Basal cortisol levels pre-op were normal. No changes were noted for pre and post op blood glucose. CONCLUSION: Pre-op screening for Cushing[apos]s is not routinely recommended unless clinically indicated. In our group with strong ACTH staining, we observed improvement in weight and blood pressure after the removal of macroadenoma, pointing toward the possible undiagnosed effect of subclinical hypercortisolism. Routine screening for patients without classic presentation should be entertained prior surgery. However our sample size is too small to draw a definite conclusion.[br][br]Nothing to Disclose: SYM, LK, BAH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 2117 322 2613 MON-745 PO22-03 Monday 2282 2012


2278 ENDO12L_MON-746 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Lack of Correlation between Cytokine Polymorphism and Cushing Disease in Turkish Patients Sema Yarman, Yeliz Ogret, Cigdem Kekik, Kursat Ozdilli, Fatma Oguz Internal Medicine, Istanbul, Turkey; Basic Sciences, Istanbul, Turkey; HealthScience, Istanbul, Turkey [bold][italic]Introduction:[/italic][/bold] Cytokines regulate many cellular responses and play regulatory roles in numerous tissues,. Especially, IL-6 has a regulatory role in the hormone secretion and growth of the anterior pituitary and is involved in adenoma pathogenesis. To test the hypothesis that a possible polymorphism of cytokine TNF-a, TGF-[beta]1, IL-10, IL-6, and IFN-[delta] might be involved in pathogenesis of Cushing[apos]s disease in our patients. We analyzed 27 (5 M, 22 F; mean age:32,07) patients and 100 healthy unrelated controls (39 M, 61 F mean age: 40.50).[br][bold][italic]Metods:[/italic][/bold] DNA was extracted from peripheric blood using a DTAB[bold]/[/bold]CTAB method. Cytokine typing: Single nucleotide gene polymorphisms, associated with the level of gene polymorphism were assessed by using polymerase chain reaction[ndash] sequence-specific primer (PCR-SSP) method by a commercially available kit (One lambda, Inc., Canoga Park, CA, USA) that contains sequence specific primers to determine the following polymorphisms: TNF-a (-308 G/A), TGF-[beta]1 (C/T codon 10, C/G codon 25), IL10 (-1082 G/A, -819 T/C, -592 A/C), IL-6 (-174 G/C), and IFN-[delta] (+874 A/T). DNA fragments corresponding to each cytokine were amplified in accordance with the manufacturer[apos]s instructions.[br][bold][italic]Results:[/italic][/bold]. Our patients have shown no significant difference regarding the distribution of IL-6, IL10, TNF-a, TGF-[beta]1, and IFN-[delta] genotype and allele polymorphism compared to controls.[br][bold][italic]Conclusion:[/italic][/bold] There is no clear evidence of the association between cytokines polymorphism and ACTH-secreting adenomas in this study. However, further molecular studies will be necessary to evaluate the hyperproduction of cytokines in the pituitary adenomas could represent an ethiopathogenic factor of cushing disease.[br][br]REFERENCES. Turnbull AV, Rivier C. Regulation of the HPA axis by cytokines. Brain Behav Immun. 1995;9(4):253-75. Kurotani R, Yasuda M, Oyama K, et al.,Expression of interleukin-6, interleukin-6 receptor (gp80), and the receptor[apos]s signal-transducing subunit (gp130) in human normal pituitary glands and pituitary adenomas. Mod Pathol. 2001;14(8):791-7. Chesnokova V, Melmed S. Minireview: Neuro-immuno-endocrine modulation of the hypothalamic-pituitary-adrenal (HPA) axis by gp130 signaling molecules. Endocrinology. 2002 May;143(5):1571-4. Haedo MR, Gerez J, Fuertes M, et al., Regulation of pituitary function by cytokines. Horm Res. 2009;72(5):266-74. Fuertes M, Gerez J, Haedo M, et al., Cytokines and genes in pituitary tumorigenesis: RSUME role in cell biology. Front Horm Res. 2010;38:1-6. Kageyama K, Kagaya S, Takayasu S, et al.,Cytokines induce NF-[kappa]B, Nurr1 and corticotropin-releasing factor gene transcription in hypothalamic 4B cells. Neuroimmunomodulation. 2010;17(5):305-13.[br][br]Nothing to Disclose: SY, YO, CK, KO, FO 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1208 322 2614 MON-746 PO22-03 Monday 2283 2012


2279 ENDO12L_MON-747 POSTER SESSION: Presentation, Treatment [amp] Outcomes of Pituitary Tumors (1:30 PM-3:30 PM) Detection of Vasoinhibin and Cathepsin D in Peripartum Cardiomyopathy and Pregnancy-Induced Hypertension Patient[apos]s Blood Asuka Hirota, Ryoujun Nakajima, Mika Suzuki, Chikage Ota, Chizuko Kamiya, Tomoaki Ikeda, Michiyo Ishida, Toshio Harigaya Meiji University, Kawasaki, Kanagawa, Japan; National Cerebral and Cardiovascular Center, Osaka, Japan It is well known that Prolactin (PRL) has modified into many variants. One of these variants, N-terminal 16kDa PRL which is cleaved by CathepsinD (CathD), has antagonistic action on native PRL. Furthermore, N-terminal cleaved hormones over than 11kDa in GH/PRL family that have anti- vasodilation and anti-angiogenesis action are named to Vasoinhibin (Vi). Vi also has been reported the possibility to induce Peripartum Cardiomyopathy (PPCM) and Pregnancy-Induced Hypertension (PIH) syndrome. PPCM is a disease of unknown etiology and exposes women to high risk of mortality after parturition. It is characterized by an acute onset of heart failure in women from the late stage of pregnancy to several months postpartum. PIH is characterized by high blood pressure and proteinuria.[br]In order to clarify these pathogenic mechanisms, we tried to detect Vi value and CathD activity in patient[apos]s blood and determine the relationship between these factors and diseases.[br]Serum or plasma samples which were collected three or four times from healthy pregnant women (control), PPCM and PIH patients at antenatal, at delivery and from one month to one year after delivery. We estimated Vi content by electrophoresis of immunoprecipitated N-terminal PRL. CathD activity in samples was also measured. Then we analysed correlations between CathD activity and brain natriuretic peptide(BNP: marker of cardiopathy) levels in PIH patients.[br]Vi content and CathD activity in PPCM and PIH patients are higher than these of control. Vi content was suppressed because PPCM patients were administrated PRL inhibitor for treatment. However, Vi content is 2.5 times higher than control, and CathD activity is also higher than control. In PIH patient, we detected a positive correlation between CathD activity and BNP. This correlation may suggest that CathD has a negative influence on the heart of pregnant women. These results seem to suggest that measurement of Vi content and CathD activity is useful for early detection of PPCM and PIH in pregnant women.[br][br]Nothing to Disclose: AH, RN, MS, CO, CK, TI, MI, TH 2012-06-25T13:30:00 Expo 2012-06-25T00:00:00 1899-12-30T13:30:00 1334 322 2615 MON-747 PO22-03 Monday 2284 2012


2280 ENDO12L_OR31-1 ORAL SESSION: Actions of Steroid Hormones [amp] Endocrine Disrupting Chemicals in Diverse Organ Systems (11:15 AM-12:45 PM) Estrogen Receptor Regulation of ERK5 and Cofilin Localization Impacts Breast Cancer Phenotypic Properties and Endocrine Resistance Zeynep Madak-Erdogan, Anna Bergamaschi, Rosa Ventrella, Hailing Lu, Benita S Katzenellenbogen University of Illinois and College of Medicine, Urbana, IL Mitogen activated protein kinases (MAPKs) are key players in relaying extracellular signals to the cell nucleus and are often overexpressed in aggressive and endocrine resistant breast cancers. Our previous studies (Madak-Erdogan et al. Mol Cell Biol 31:226,2011) revealed that one such kinase, ERK2, was recruited to distal enhancers of estradiol(E2)-regulated genes together with estrogen receptor alpha (ER[alpha]), resulting in proper control of gene expression and cell proliferation programs. In the current study, we identified ERK5, a close relative of ERK2, as a factor that is recruited to the transcription start site (TSS) of E2-stimulated genes. Use of dominant negative and constitutively active forms, as well as small molecule inhibitors of ERK5 and MEK5, revealed that activation of ERK5 by upstream kinase MEK5 and activity of ERK5 itself was required for recruitment of the kinase to the chromatin. Immunofluorescence studies demonstrated that ERK5 localized to transcription factories in the nucleus upon cell treatment with E2. Recruitment of ERK5 to the TSS of genes was necessary for Paf1 (Polymerase associated factor) recruitment to the RNA Pol II complex and transcription elongation. Inhibitors of ERK5 completely abrogated E2-dependent cell proliferation and colony formation for various ER[alpha]-positive breast cancer cell lines studied. In contrast to what we observed in ER[alpha]-positive cells, in ER[alpha]-negative breast cancer cell lines we did not detect any nuclear ERK5; instead, we found ERK5 localized to the actin cytoskeleton, especially to sites where substantial remodeling takes place. Upon re-expression of ER[alpha], we observed re-localization of ERK5 to the nucleus and a decrease in actin reorganization associated with reduced cell motility and invasion. Abrogation of ER[alpha] expression using siRNA in ER[alpha]-positive cell lines caused loss of nuclear ERK5 and relocalized this kinase to the actin cytoskeleton. Our studies with tamoxifen resistant cell lines showed that ERK5 localization to the nucleus decreased and localization to the actin cytoskeleton increased as resistance progressed, suggesting a correlation between ER[alpha] activity, ERK5 localization and motility of breast cancer cells. Our data reveal that ER[alpha] elicits nuclear localization of ERK5, diminishing ERK5 localization to regions of high actin remodeling, thereby possibly accounting for the generally lower metastatic potential that is characteristic of many ER[alpha]-positive breast cancer cells.[br][br]Sources of Research Support: NIH, T32 ES007326 and P50 AT006268, and from The Breast Cancer Research Foundation.[br][br]Nothing to Disclose: ZM-E, AB, RV, HL, BSK 2012-06-26T11:15:00 320 2012-06-26T00:00:00 1899-12-30T11:15:00 852 374 2711 OR31-1 OR24-01 Tuesday 2285 2012


2281 ENDO12L_OR31-2 ORAL SESSION: Actions of Steroid Hormones [amp] Endocrine Disrupting Chemicals in Diverse Organ Systems (11:15 AM-12:45 PM) Estrogen Via ER[beta] Reciprocally Regulates Class I and II HDACs To Prevent Cardiomyocyte Hypertrophy Ali Pedram, Mahnaz Razandi, Ellis R Levin University of California, Irvine, CA; Long Beach VA Medical Center, Long Beach, CA Cardiac hypertrophy arises from poorly controlled hypertension and ischemic heart disease, progressing to dilation and cardiac failure as the leading cause of death in Americans.We have published that signaling from the membrane ERbeta receptor prevents cardiac hypertrophy and cardiac fibrosis, in-vitro and in-vivo. One important mechanism for development of the cardiac pathology involves Class I and Class II histone de-acetylases (HDAC) that promote or inhibit cardiac hypertrophy, respectively. We determined that Angiotensin II (AngII) and Endothelin-1 (ET-1), important factors that promote human cardiac hypertrophy, induced new protein synthesis of Class I HDAC2 in freshly isolated neonatal rat cardiomyocytes and stimulated HDAC2 gene transcription (qPCR). The pro-hypertrophic HDAC2 protein is activated by phosphorylation in the nucleus. AngII stimulated CK2 (creatine kinase) activity and its nuclear translocation where it phosphorylated HDAC2 at Ser392. All these effects of AngII/ET-1 were significantly blocked by 17-beta-estradiol (E2) or the ERbeta agonist DPN (each at 10nM). In contrast, AngII and ET-1 inhibited the synthesis of Class II HDACS 4 and 5 (anti-hypertrophic) in PKCdelta and protein kinase D (PKD)-linked fashion. E2 or DPN prevented AngII and ET-1-inhibited synthesis of these HDAC proteins, and the estrogenic compounds[apos] effects were reversed by siRNA to ERbeta but not ERalpha siRNA. AngII also caused increased phosphorylation of Serine259/Serine498 in HDAC5 through PKCdelta activation, resulting in export of the anti-hypertrophic HDAC from nucleus into cytoplasm, thus contributing to hypertrophy. PKCdelta activation, Serine259 phosphorylation, and nuclear export of HDAC5 were all inhibited by E2 or DPN. HDAC4 phosphorylation at Ser632 is necessary for nuclear export and was stimulated by AngII in both a CAM kinase and PKD dependent fashion, inhibited by E2/DPN. Thus, AngII stimulated a linear pathway, involving CAM kinase II-PKCdelta-PKD, inhibited by E2 and DPN. AngII signaling to the decreased production and nuclear exclusion of anti-hypertrophic HDACs 4 and 5, but increased production and nuclear localization of the pro-hypertrophic HDAC2 in cardiomyocytes was suppressed by E2 and DPN. This reciprocal modulation of pro and anti-hypertrophic HDACs is a novel mechanism for prevention of cardiomyocyte hypertrophy in general, and contributes to the anti-hypertrophic functions of E2/ERbeta.[br][br]Nothing to Disclose: AP, MR, ERL 2012-06-26T11:30:00 320 2012-06-26T00:00:00 1899-12-30T11:30:00 23 374 2712 OR31-2 OR24-01 Tuesday 2286 2012


2282 ENDO12L_OR31-3 ORAL SESSION: Actions of Steroid Hormones [amp] Endocrine Disrupting Chemicals in Diverse Organ Systems (11:15 AM-12:45 PM) Genome-Wide Analysis of Androgen Receptor Cistromes in Mouse Prostate and Kidney, Two Androgen-Responsive Tissues Paivi Pihlajamaa, Biswajyoti Sahu, Viljami Aittomaki, Sampsa Hautaniemi, Olli A Janne Biomedicum Helsinki, University of Helsinki, Helsinki, Finland; Biomedicum Helsinki, University of Helsinki, Helsinki, Finland Androgens regulate the expression of their target genes in a tissue-specific manner. The effects of androgens are mediated through the androgen receptor (AR), a ligand inducible transcription factor that binds to androgen response elements (ARE) in the regulatory regions of the target genes. Genome-wide mapping of androgen receptor binding sites (ARB) in prostate cancer cell lines has revealed that the majority of AR binding events occur far away from transcription start sites of the genes regulated by androgens. To study genome-wide AR binding to chromatin [italic] in vivo[/italic], we analyzed AR cistromes by ChIP-sequencing in two androgen-responsive murine tissues, the prostate and the kidney. ChIP was performed after a two-hour testosterone exposure of castrated male mice. The ChIP-seq data from two biological replicates on both tissues revealed the presence of 6,618 and 2,965 ARBs in prostate and kidney, respectively (false discovery rate [lt] 2%). AR-binding profiles were distinct in these two tissues, in that less than one-fourth of the ARBs in kidney overlapped with the ARBs in prostate. [italic] De novo[/italic] motif search revealed that a canonical ARE sequence was enriched among the ChIP-seq peaks both in prostate and in kidney. However, enriched motifs for other [italic] cis[/italic]-regulatory elements adjacent to ARBs show differences between the two tissues. To determine androgen-regulated transcripts in murine prostate and kidney, gene expression profiling was performed on tissues of castrated male mice after a three-day testosterone or vehicle treatment. With a fold-change [gt] 1.5 and p-value [lt] 0.05 cut-offs, we identified 587 and 804 differentially expressed genes in prostate and kidney, respectively. In agreement with the ARBs, most androgen-regulated transcripts were tissue-specific, as only 9% of up-regulated and 15% of down-regulated transcripts were common in prostate and kidney. Of note, ARBs were on many occasions tissue-specific even for genes that were up-regulated by androgens in both tissues. In summary, androgen signaling in mouse prostate and kidney, two androgen-responsive tissues, involves the use of distinct chromatin AR-binding sites [italic] in vivo[/italic], which results in tissue-specific transcription programs.[br][br]Nothing to Disclose: PP, BS, VA, SH, OAJ 2012-06-26T11:45:00 320 2012-06-26T00:00:00 1899-12-30T11:45:00 1197 374 2713 OR31-3 OR24-01 Tuesday 2287 2012


2283 ENDO12L_OR31-4 ORAL SESSION: Actions of Steroid Hormones [amp] Endocrine Disrupting Chemicals in Diverse Organ Systems (11:15 AM-12:45 PM) Estrogenic Endocrine-Disrupting Chemicals Reprogram Human Prostate Stem/Progenitor Cells Wen-Yang Hu, Guang-Bin Shi, Dan-Ping Hu, Gail S Prins University of Illinois at Chicago, Chicago, IL Estrogen reprogramming of the rodent prostate by developmental exposures results in permanent alterations in structure and gene expression, leading to prostate neoplasia with aging. Endocrine disrupting chemicals (EDCs) have been similarly linked to increased prostate cancer risk in men. Recently, we documented that human prostate epithelial stem and early-stage progenitor cells express ER[alpha], ER[beta] and GPR30 and show proliferative responses to estradiol-17[beta] (E2) suggesting that they are direct estrogen targets (Hu et al, Endocrinology, 2011). Since stem cells are potential targets of cancer initiation, we hypothesize that estrogens and EDCs may influence the development and progression of prostate cancer through stem/progenitor cell reprogramming. The present study sought to examine the direct effects of EDCs on human prostate stem cell self-renewal, progenitor amplification and differentiation using a novel prostasphere assay. Prostaspheres (PS) of stem/progenitor cells were established in 3-D culture from disease-free human prostate epithelial cell primary cultures. Similar to E2 effects, exposure to bisphenol A (BPA) at doses ranging 10nM - 10mM significantly stimulated PS number and size at day 7 indicating increased stem cell self-renewal and progenitor cell amplification. Hoechst exclusion with flow cytometry analysis confirmed an increased stem-cell side population with BPA treatments in parental primary cells. In contrast, sodium arsenite (NaAsO2) markedly decreased PS number and size in a dose-dependent manner (0.5, 5, 50 mM). While BPA did not influence ER[alpha], ER[beta] and GPR30 expression, PS exposure to NaAsO2 markedly decreased ER[alpha] and ER[beta] expression. Treatment of PS with 10nM retinoic acid (RA) drives PS differentiation as monitored morphologically and by gene expression analysis. Exposure to 200 nM BPA suppressed RA-induced PS differentiation with reduced ER[beta], AR, FOXA1 and HOXB13 expression. In contrast, the differentiation response to NaAsO2 was divergent with a dose-dependent decrease in AR and HOXB13 expression but an increase in NKX3.1 expression suggesting differentiation perturbation in progenitor cells. Together, these findings indicate that estrogens and EDCs have diverse effects on human prostate stem/progenitor cell proliferation and differentiation perhaps mediated through distinct ERs. Since stem cell alterations are long lasting, such perturbations may underpin prostate cancer risk throughout life as a function of EDC exposures.[br][br]Hu WY et al, Endocrinology 2011;152:2150.[br][br]Sources of Research Support: NIH grants ES015584 and ES018758.[br][br]Nothing to Disclose: W-YH, G-BS, D-PH, GSP 2012-06-26T12:00:00 320 2012-06-26T00:00:00 1899-12-30T12:00:00 2075 374 2714 OR31-4 OR24-01 Tuesday 2288 2012


2284 ENDO12L_OR31-5 ORAL SESSION: Actions of Steroid Hormones [amp] Endocrine Disrupting Chemicals in Diverse Organ Systems (11:15 AM-12:45 PM) Prepubertal Exposure to Manganese (Mn) Increases Endogenous Estradiol (E[sub]2[/sub]), Resulting in Accelerated Mammary Gland Development and Hyperplasic Lesions in Adult Female Rats Robert K Dearth, Jill K Hiney, Vinod K Srivastava, Jesse R Rivera, Kelly Scribner, Weston Porter, W Les Dees University of Texas-Pan American, Edinburg, TX; Texas A[amp]M University, College Station, TX Early age at menarche results in a 10-20% increased risk of developing breast cancer. Strong evidence suggests that exposure to environmental substances that regulate estrogenic pathways play a role in linking early pubertal onset to increased breast cancer incidence. We have recently shown that prepubertal exposure to low levels of manganese (Mn) induces precocious puberty in rats, via a centrally-mediated action, resulting in elevated puberty-related hormones including estradiol (E[sub]2[/sub]). However, the effect of Mn-induced precocious puberty on mammary development has not been determined. Therefore, we assessed the ability of prepubertal Mn exposure to advance normal breast development and alter E[sub]2[/sub] driven pathways known to increase the gland[apos]s susceptibility to tumors. Sprague Dawley females were gavaged daily with either 10mg/kg MnCl[sub]2[/sub] or saline (controls) starting on post natal day (PND) 12 through PND 30. Then, blood and mammary glands (MGs) were collected from half of the treated and controls groups on PND 30 and the other half on PND 120. As expected, serum E[sub]2[/sub] levels were elevated (p[lt]0.05) in the Mn-treated group compared to controls on PND 30. Importantly, Ki67 staining revealed that Mn exposure increased (p [lt] 0.001) mammary epithelial cell proliferation compared to controls. In addition, a higher (p [lt]0.05) number of alveolar buds and lobular type-1 structures were observed in the Mn-treated females. Analysis of MGs on PND 120 revealed that MnCl[sub]2[/sub]-treated animals continued to have increased (p[lt] 0.001) cellular proliferation compared to controls. Importantly, levels of Mn were not elevated in these MGs. Western blot and immunocytochemical analysis revealed that ER[alpha], SP1, AP2[alpha] and Kiss-1were elevated (p[lt]0.05-0.01) in MG epithelial cells on PND120 in Mn-treated animals. These differences were not observed on PND 30. Surprisingly, hyperplasic ductal structures as well as mis-localization of keratin-14 were observed in Mn-treated animals at PND 120. Overall, this is the first study to show that prepubertal exposure to Mn accelerates MG growth resulting in hyperplasic lesions in adult females. Furthermore, the alterations in signaling we observed are similar to the protein signature of the E[sub]2[/sub] driven human breast cancer cell line MCF-7. Thus, we have created an animal model that will provide important information regarding environmentally regulated precocious puberty and the subsequent risk of developing breast cancer.[br][br]Sources of Research Support: ESO13143 to WLD.[br][br]Nothing to Disclose: RKD, JKH, VKS, JRR, KS, WP, WLD 2012-06-26T12:15:00 320 2012-06-26T00:00:00 1899-12-30T12:15:00 998 374 2715 OR31-5 OR24-01 Tuesday 2289 2012


2285 ENDO12L_OR31-6 ORAL SESSION: Actions of Steroid Hormones [amp] Endocrine Disrupting Chemicals in Diverse Organ Systems (11:15 AM-12:45 PM) Bisphenol-A Exposure In Utero Programs Adult Uterine Estrogen Responsiveness Elisa M Jorgensen, Yuping Zhou, Hugh S Taylor Yale University School of Medicine, New Haven, CT Introduction: Bisphenol-A is a ubiquitous estrogen-like endocrine-disrupting compound. In utero exposure to BPA is linked to female reproductive disorders including endometrial hyperplasia, endometriosis, fibroids and breast cancer. Estrogens play a role in the etiology of each of these conditions. Here we hypothesize that BPA exposure in utero results in altered developmental programming of estrogen responsive gene expression, and that the mechanism of BPA-induced reprogramming is epigenetic change to DNA methylation.[br]Methods: 8 pregnant CD-1 mice were treated with BPA (5 mg/kg/d) or vehicle via osmotic minipump on d9-18 of gestation. At 2 wks, uteri of half the offspring were isolated and RNA was extracted. At 6 wks, remaining females were oophorectomized, treated with 300 ng E2 or vehicle, and uteri were removed for RNA and DNA isolation. Total RNA was labeled and hybridized to a BeadChip WG-6 expression microarray (Illumina). Genes showing statistically significant change ([gt] 2-fold) versus control were verified using real time RT-PCR. DNA was enriched using MeDIP, then labeled and hybridized to a 720K CpG promoter methylation array (Nimblegen).[br]Results: At 2 wks, global gene expression was remarkably similar among control and BPA-exposed groups. Of 45,000 genes screened, only 18 showed 2-fold or greater changes in expression. After estrogen exposure at puberty (8 wks), the expression profile was markedly changed. At baseline, 365 genes showed altered expression in BPA-exposed offspring. Another 316 genes showed altered E2 response in BPA-treated offspring; this included several genes not previously regulated by estrogen (Fzd10, Gdf10), or demonstrating an exaggerated response to E2 treatment (Ret, Wif1, S100a8). Of genes with altered expression at puberty, 208 also showed aberrations to the normal pattern of methylation, including changes to at least 14 genes with known EREs (G6pdx, Stat5a, Scd1).[br]Conclusions: Altered gene expression was observed in uteri of mice exposed to BPA prenatally; however, these differences occur only after endogenous estrogen exposure at puberty or with E2 treatment. Gene-environment interactions driven by BPA alter the normal developmental programming of estrogen responsive gene expression in the uterus. Estrogen hyperresponsiveness is a potential mechanism to explain increased incidence of estrogen-related disorders seen after exposure to endocrine disruptors. Many of these changes may be mediated by altered ERE methylation.[br][br]Nothing to Disclose: EMJ, YZ, HST 2012-06-26T12:30:00 320 2012-06-26T00:00:00 1899-12-30T12:30:00 1136 374 2716 OR31-6 OR24-01 Tuesday 2290 2012


2286 ENDO12L_OR32-1 ORAL SESSION: Novel Insights in GHRH-GH-IGF1 Biology (11:15 AM-12:45 PM) Novel Actions of Growth Hormone (GH) on Macrophage (M[Phi]) Polarization Result in Paradoxical Effects of GH on Glucose Homeostasis Chunxia Lu, Anil K Pasupulati, Carey Lumeng, Yong Fan, Mark A Sperling, Ram K Menon University of Michigan, Ann Arbor, MI; University of Pittsburgh, Pittsburgh, PA In diet-induced obesity (DIO), adipose tissue (AT) M[Phi]s (ATM) are skewed towards the classically activated pro-inflammatory M1 (F4/80+, CD11c+) phenotype as opposed to the alternative activated anti-inflammatory M2 (F4/80+, CD206+) phenotype that predominate in the lean state. M1 M[Phi]s secrete pro-inflammatory cytokines implicated in DIO-associated insulin resistance (IR). The role of endocrine factors in the regulation of M[Phi] polarization (MP) in DIO is not fully understood.[br]In this study, we evaluated the effect of GH on MP using a M[Phi]-specific GH receptor knockout mouse model (MacGHR KO). MacGHR KO [amp] control mice were fed high fat diet (45 kcal% x 18 wks) prior to isolation/purification of M[Phi]-enriched AT stromal vascular fraction (SVF). Flow cytometry revealed increased M1/M2 ratio in MacGHR KO, suggesting that under normal circumstances GH has a negative effect on M1 and/or positive effect on M2 M[Phi] polarization in DIO. In response to LPS, M1 MP (expression of M1 markers iNOS2, IL1[beta], [amp] IL12) was enhanced in MacGHR KO bone-marrow-derived M[Phi]s (BMDM). Consistent with enhanced M1 MP, expression of pro-inflammatory cytokines (IL6, TNF[alpha], IL1[beta] [amp] osteopontin) was increased in MacGHR KO SVF. Crown-like-structures, typically associated with DIO-associated chronic inflammation in AT, was increased in MacGHR KO AT. [italic]Ex vivo[/italic] stimulation of isolated AT yielded decreased insulin-dependent AKT phosphorylation in MacGHR KO. [italic]In vivo[/italic], the pro-inflammatory changes in AT were accompanied by impaired glucose tolerance (via ipGTT) and increased IR (via ITT) in MacGHR KO. These findings contravene the current dogma that GH is a diabetogenic hormone that increases IR and that loss of GH action, such as in MacGHR KO, should only [underline]decrease[/underline] IR in DIO. NF-kB is known to be essential for M1 MP. Western blotting revealed increased levels of phosphorylated NF-kB in MacGHR KO SVF compared to controls, suggesting that GH has a negative effect on NF-kB activation. This inhibitory effect of GH on NF-kB activation was also observed in J774.1 M[Phi]s. We conclude that in DIO, lack of GH action on the M[Phi] exerts a deleterious effect on glucose homeostasis by accentuating ATM inflammation with increased NF-kB activation and M1 M[Phi] MP. These results in mice provide novel insights into GH[apos]s role in the maintenance of metabolic homeostasis in DIO and support the possibility that administration of GH could have salutary effects on DIO-associated chronic inflammation and IR in humans.[br][br]Nothing to Disclose: CL, AKP, CL, YF, MAS, RKM 2012-06-26T11:15:00 372 2012-06-26T00:00:00 1899-12-30T11:15:00 973 375 2717 OR32-1 OR18-01 Tuesday 2291 2012


2287 ENDO12L_OR32-2 ORAL SESSION: Novel Insights in GHRH-GH-IGF1 Biology (11:15 AM-12:45 PM) Insulin-Like Growth Factor 1 (IGF1) Encodes Memory of Past Food Intake by Regulating Functional Capacity of the Gastric Neuromuscular Apparatus Yoshitaka Toyomasu, Yujiro Hayashi, Andrea Lorincz, Michael R Bardsley, Laura N Popko, David L Young, Jessica E Mason, Simon J Gibbons, David R Linden, Joseph H Szurszewski, Gianrico Farrugia, Tamas Ordog Mayo Clinic, Rochester, MN [underline]Background [amp] Aims:[/underline] Gastric symptoms and delayed gastric emptying (gastroparesis) are common in anorexia nervosa and secondary protein-energy malnutritions and interfere with nutritional rehabilitation (1). Conversely, gastrointestinal (GI) motility disorders arising from abnormalities in gastric smooth muscle cells (SMC), enteric neurons (EN) and interstitial cells of Cajal (ICC) are often accompanied by reduced food intake (2). Recently we found that chronic dietary restriction (CDR) in female mice led to gastroparesis and reduced EN and ICC networks (3). ICC and nerve fibers were also depleted in patients with severe chronic weight loss. CDR reduces IGF1, which plays an important role in the maintenance of SMC and ICC (4, 5). Here we studied whether weight gain after CDR is limited by gastric cell loss and whether it can be restored by IGF1. [underline]Methods:[/underline] Female BALB/c mice aged 4, 8 or 13 weeks (n=120 in 3 cohorts) were exposed to CDR adjusted daily to prevent natural body weight (bwt) gain for 170-224 days. Subsets of mice were allowed ad-libitum access to food starting at 0 (AL controls), 96, 83 or 152 days of CDR (refed groups; RF) and/or treated with R3 LONG recombinant human IGF1 (150 [mu]g/kg sc. BID) starting with CDR or refeeding. Solid gastric emptying was measured by a [sup]13[/sup]C breath test. Estrous cyclicity was assessed by vaginal cytology. Blood glucose, ketones, serum malondialdehyde, insulin, IGF1 and corticosterone were measured by commercial kits. Gene expression specific for ICC, SMC and EN was studied by Western immunoblotting. [underline]Results:[/underline] AL mice displayed age-dependent, exponential rise in bwt. CDR mice had lower ketone ([italic]P[/italic]=0.002) and IGF1 levels ([italic]P[/italic]=0.016) and 81% were in anestrus. Solid gastric emptying was delayed ([italic]P[/italic][lt]0.016). CDR significantly reduced the cell type markers Kit (ICC), Myh11 (SMC), Uchl1 and Nos1 (EN). Refeeding of mice whose CDR was started at 4 or 8 weeks of age led to reduced food intake vs. AL controls and bwt curves running 10-15% below those of AL mice ([italic]P[/italic][lt]0.001); whereas in older mice, bwts quickly normalized. In the young cohorts, refeeding caused a further delay in gastric emptying ([italic]P[/italic][lt]0.001). IGF1 normalized gene expression and gastric emptying in both CDR and RF mice and restored bwts in RF mice. [underline]Conclusions:[/underline] CDR reduces functional capacity of the GI neuromuscular apparatus likely through the dystrophic effects of low circulating IGF1. In young mice these changes persist after refeeding but can be minimized by IGF1 treatment.[br][br](1) Hadley SJ [amp] Walsh BT, Curr Drug Targets CNS Neurol Disord 2003; 2:1. (2) Parkman HP et al., Gastroenterology 2011; 141:486. (3) Lorincz A et al., Neurogastroenterol Motil 2008; 20(Suppl 1):80. (4) Horvath VJ et al., Gastroenterology 2006; 130:759. (5) Bardsley MR et al., Gastroenterology 2010; 139:942.[br][br]Sources of Research Support: NIH Grant DK68055.[br][br]Nothing to Disclose: YT, YH, AL, MRB, LNP, DLY, JEM, SJG, DRL, JHS, GF, TO 2012-06-26T11:30:00 372 2012-06-26T00:00:00 1899-12-30T11:30:00 1877 375 2718 OR32-2 OR18-01 Tuesday 2292 2012


2288 ENDO12L_OR32-3 ORAL SESSION: Novel Insights in GHRH-GH-IGF1 Biology (11:15 AM-12:45 PM) Leptin Increases IGFBP-2 in Skeletal Muscle by Central and Peripheral Mechanisms Steven W Yau, Belinda A Henry, Vincenzo C Russo, Glenn K McConell, Iain J Clarke, George A Werther, Matthew A Sabin Murdoch Children[apos]s Research Institute, Royal Children[apos]s Hospital, Melbourne, Australia; University of Melbourne, Melbourne, Australia; Monash University, Melbourne, Australia; Victoria University, Melbourne, Australia [bold]Introduction:[/bold] Insulin-like growth factor (IGF) binding protein 2 (IGFBP-2) an IGF binding protein with IGF-dependent and IGF-independent actions, may represent a critical link between weight status and insulin sensitivity [sup](1,2,3)[/sup], such that reduced levels are associated with obesity and insulin resistance. Recently, IGFBP-2 has been shown to be regulated by leptin [sup](4)[/sup], but the sites and mechanisms of this regulation are not known.[br][bold]Aims:[/bold] To investigate effects of leptin on IGFBP-2 mRNA expression in skeletal muscle (SM), liver, subcutaneous (s/c) and visceral adipose tissue, and compare indirect (brain-mediated) and/or direct effects of leptin on IGFBP-2 mRNA expression and protein secretion in SM.[br][bold]Methods:[/bold] A) Sheep were randomised to receive infusions of either human recombinant leptin in artificial cerebrospinal fluid (aCSF)(50[mu]g/h; n=4) or vehicle (aCSF: 60[mu]l/h; n=8) via lateral ventricular (LV) cannulae for 8 days. A further control group were pair-fed to the leptin-infused animals (n=4). At the end of the infusion period, the animals were euthanased and samples of SM, liver, s/c and visceral fat were collected and rapidly frozen in liquid nitrogen. IGFBP-2 mRNA expression was measured by qPCR. B) [italic]In-vitro[/italic] cultures of human skeletal myotubes (HSM) were treated with either 50 or 100nM leptin for 30 min, 24 or 48h (n=4). IGFBP-2 mRNA expression (qPCR) and IGFBP-2 levels (ELISA) were measured.[br][bold]Results:[/bold] A) Sheep receiving ICV leptin exhibited a 15 fold (p[lt]0.001) increase in IGFBP-2 mRNA expression in SM (vs control or pair-fed animals), while IGFBP-2 expression was increased 2.5 fold (p[lt]0.01) in liver, 1.9 fold (p[lt]0.01) in s/c fat and 2.8 fold (p[lt]0.01) in visceral fat. B) Direct leptin dosing in HSM increased IGFBP-2 expression at 50nM by 1.8 fold (p[lt]0.05) and at 100nM by 2.2 fold (p[lt]0.01) at 30mins. By 24 and 48h, treatment with 100nM leptin increased IGFBP-2 mRNA expression by 7.9 fold (p[lt]0.001) and 10.3 fold (p[lt]0.001), respectively, while secreted IGFBP-2 in conditioned media increased 15 fold (p[lt]0.05) and 34 fold (p[lt]0.001), respectively.[br][bold]Conclusion:[/bold] These data suggest that leptin increases IGFBP-2 expression in SM through both central (brain-mediated) and peripheral mechanisms. IGFBP-2 may represent a critical mediator by which leptin regulates insulin sensitivity, with potential implications for the insulin resistance of obesity.[br][br](1) Sabin, MA et al. IGFBP-2 at the interface of growth and metabolism-implications for childhood obesity. Pediatric Endocrinology Reviews. 2011;8(4):382-93. (2) Sabin, MA et al. Dietary monounsaturated fat in early life regulates IGFBP-2 [ndash] implications for fat mass accretion and insulin sensitivity. Obesity 2011;19(12):2374-81. (3) Wheatcroft, SB et al. IGF-binding protein-2 protects against the development of obesity and insulin resistance. Diabetes 2007;56:285-94. (4) Hedbacker, K et al. Antidiabetic effects of IGFBP2, a leptin-regulated gene. Cell Metabolism 2010;11:11-22.[br][br]Nothing to Disclose: SWY, BAH, VCR, GKM, IJC, GAW, MAS 2012-06-26T11:45:00 372 2012-06-26T00:00:00 1899-12-30T11:45:00 1185 375 2719 OR32-3 OR18-01 Tuesday 2293 2012


2289 ENDO12L_OR32-4 ORAL SESSION: Novel Insights in GHRH-GH-IGF1 Biology (11:15 AM-12:45 PM) Increased Expression of Fibroblast Growth Factor 21 (FGF21) during Chronic Undernutrition Causes Growth Hormone Insensitivity in Chondrocytes by Inducing Leptin Receptor Overlapping Transcript (LEPROT) Expression Tal Grunwald, Shufang Wu, Francesco De Luca St Christopher[apos]s Hospital for Children, Philadelphia, PA [bold]Background[/bold][br]During calorie restriction in mice, increased expression of Fibroblast Growth Factor 21 (FGF21) causes growth attenuation and Growth Hormone (GH) insensitivity. Previous evidence also indicates that fasting-associated increased expression of LEPROT (a protein involved in the regulation of intracellular protein trafficking) reduces GH receptor abundance on the hepatocyte cell membrane. Based on these findings, we hypothesized that FGF21 causes GH insensitivity through its functional interaction with LEPROT.[br][bold]Results[/bold][br]After 4 weeks of 50 % food restriction, LEPROT mRNA expression (by real-time PCR) in the tibial growth plate of wild-type (WT) mice was increased compared to WT mice fed ad lib (+ 243 %, p[lt]0.05). In contrast, LEPROT mRNA expression in food-restricted and fed ad-lib [italic]Fgf21[/italic] knock-out mice was similar.[br]We then isolated chondrocytes from fetal (dpc20) mouse metatarsal growth plates and transfected them with control short interfering RNA (siRNA) or LEPROT siRNA.[br][underline]In cells transfected with control siRNA[/underline], 10 ng/ml GH increased [sup]3[/sup]H-thymidine incorporation (+158 % of untreated cells, p[lt]0.01) and collagen X mRNA expression (+392 %, p[lt]0.05), with both effects being prevented by 100 ng/ml rhFGF21 (GH+rhFGF21 vs. GH alone: [sup]3[/sup]H-thymidine incorporation: +118 % vs. +158 %, p[lt]0.05. Collagen X expression: +211 % vs. +392 %, p[lt]0.05). GH also induced IGF-1 mRNA expression (500 % of untreated cells, p[lt]0.01), while co-treatment with rhFGF21 neutralized this effect (GH+FGF21 vs. GH alone: +294 % vs. +500 %, p[lt]0.05). In addition, rhFGF21 decreased [[sup]125[/sup]I]-GH binding (GH+FGF21 vs. GH alone: + 83 % vs. +127 %, p[lt]0.01).[br][underline]In LEPROT siRNA-transfected cells[/underline], GH also stimulated thymidine incorporation (+168 % of untreated cells, p[lt]0.01) and collagen X mRNA expression (+407%, p[lt]0.01). In contrast to its effects in control siRNA-transfected cells, in LEPROT siRNA-transfected cells rhFGF21 did not prevent the GH stimulatory effects on thymidine incorporation (GH+FGF21 vs. GH alone, +144 % vs. + 148 %) or Collagen X expression (+ 375 % vs. + 365 %). Furthermore, rhFGF21 did not prevent [[sup]125[/sup]I]-GH binding (GH+FGF21 vs. GH alone: + 121 % vs. + 106 %) or the GH stimulatory effect on IGF-1 mRNA expression (GH+FGF21 vs. GH alone: +462 % vs. +443 %).[br][bold]Conclusions[/bold][br]Our findings suggest that the increased expression of FGF21 during chronic undernutrition inhibits GH action on chondrocytes by activating LEPROT, which, in turn, reduces GH receptor abundance on the cell membrane.[br][br]Nothing to Disclose: TG, SW, FDL 2012-06-26T12:00:00 372 2012-06-26T00:00:00 1899-12-30T12:00:00 243 375 2720 OR32-4 OR18-01 Tuesday 2294 2012


2290 ENDO12L_OR32-5 ORAL SESSION: Novel Insights in GHRH-GH-IGF1 Biology (11:15 AM-12:45 PM) Changes in IGF-I Bioactivity Do Not Parallel Changes in Total IGF-I during GH Treatment of GH-Deficient Adults Aimee J Varewijck, Steven WJ Lamberts, Sebastiaan JCMM Neggers, Leo J Hofland, Joseph AMJL Janssen Erasmus Medical Center, Rotterdam, Netherlands [bold]Context[/bold] Previously we demonstrated that IGF-I bioactivity offers advantages in the diagnostic evaluation of adult GHD. It is unknown whether IGF-I bioactivity can be used to monitor GH therapy.[br][bold]Aim[/bold] To investigate the value of IGF-I bioactivity for monitoring the effect of GH therapy.[br][bold]Methods[/bold] 106 patients (54 m; 52 f) diagnosed with GHD by GH-provocative tests were included; 22 were GH-naive, 84 were already on GH treatment and discontinued therapy 4 weeks before establishing baseline values. IGF-I bioactivity was determined by the IGF-I KIRA assay, total IGF-I by immunoassay (Immulite) in fasting blood samples collected at baseline and after 12 months of GH therapy. GH doses were titrated to achieve total IGF-I within the normal range.[br][bold]Results[/bold] Total IGF-I increased from 8.2[plusmn]0.5 nmol/l (Z-score -2.2SD) at baseline to 14.9[plusmn]0.7 nmol/l (Z-score -0.6SD) at 12 months. IGF-I bioactivity increased from 116[plusmn]6 pmol/l (Z-score -2.6SD) to 179[plusmn]11 pmol/l (Z-score -1.8SD). At 12 months the mean GH dose was 0.20 mg/day (range: 0.05-2.1).[br]At baseline, total IGF-I was within the normal range in 37% of patients, compared to 22% for IGF-I bioactivity. After 12 months these percentages increased to 81% and to 51% respectively (87% vs. 53%, respectively, after excluding oral estrogen use). After 12 months, IGF-I bioactivity was below normal in more than 40% of patients in whom total IGF-I had normalized.[br]At baseline, with an increasing number of additional pituitary deficits, total IGF-I was less frequently within the normal range (additional deficits: 1-3; 44% vs. [ge]3; 35%). For IGF-I bioactivity this percentage was independent of the number of additional deficits (23% vs. 21%, respectively). After GH therapy, with an increasing number of additional deficits, total IGF-I was more frequently within the normal range (69% vs. 89%). For IGF-I bioactivity this percentage was substantially lower and independent of the number of additional deficits (54% vs. 48%). Patients with [ge]3 additional deficits were older and heavier than patients with [lt]3 additional deficits (p[lt]0.001 and p=0.04, respectively).[br][bold]Conclusion[/bold] During 12 months of GH treatment changes in IGF-I bioactivity did not parallel changes in total IGF-I. In a considerable proportion of patients IGF-I bioactivity was subnormal despite normalization of total IGF-I. Since IGF-I bioactivity probably better correlates with biological activity, our study suggests that total IGF-I is not a reliable marker of exogenous GH activity.[br][br]Sources of Research Support: J.A.M.J.L. Janssen has received an unrestricted grant from Novo Nordisk A/S (Alphen aan de Rijn, the Netherlands).[br][br]Nothing to Disclose: AJV, SWJL, SJCMMN, LJH, JAMJLJ 2012-06-26T12:15:00 372 2012-06-26T00:00:00 1899-12-30T12:15:00 1457 375 2721 OR32-5 OR18-01 Tuesday 2295 2012


2291 ENDO12L_OR32-6 ORAL SESSION: Novel Insights in GHRH-GH-IGF1 Biology (11:15 AM-12:45 PM) Metabolic Effects of a Growth Hormone-Releasing Factor in Obese Subjects with Reduced Growth Hormone Secretion: A Randomized, Double-Blind, Placebo-Controlled Study Hideo Makimura, Meghan N Feldpausch, Alison M Rope, Linda C Hemphill, Martin Torriani, Hang Lee, Steven K Grinspoon Massachusetts General Hospital, Boston, MA; Massachusetts General Hospital, Boston, MA; Massachusetts General Hospital, Boston, MA; Massachusetts General Hospital, Boston, MA [bold]Background[/bold]: Obesity is associated with reduced GH secretion, and increased cardiovascular disease (CVD) risk.[br][bold]Methods[/bold]: To assess the effects of augmenting GH secretion on body composition and CVD risk indices, 60 obese patients (mean BMI 38[plusmn]0.6 kg/m[sup]2[/sup]) with reduced GH (peak GH to GHRH-arginine testing [le]9 [micro]g/L) were randomly assigned to a GHRH[sup]1-44 [/sup]analogue, tesamorelin 2 mg SC QD, or placebo for 12 months. Treatment effect was determined by longitudinal linear mixed effects modeling. Dose reductions, with parallel dummy changes to placebo-treated subjects, were made by an independent physician if IGF-1 increased above normal.[br][bold]Results:[/bold] Body composition and demographic parameters were similar at baseline. 83% of subjects completed 6 months and 62% completed 12 months, without differences in discontinuation rates between the groups. IGF-1 increased (86[plusmn]21 vs. -6[plusmn]8 [micro]g/L, tesamorelin vs. placebo; treatment effect [95% CI]: 92 [52,132] [micro]g/L; P[lt]0.0001). Abdominal visceral adipose tissue (VAT) (-16[plusmn]9 vs.19[plusmn]9 cm[sup]2[/sup]; -35 [-58,-12] cm[sup]2[/sup]; P=0.003) (net treatment effect of -19% relative to placebo), carotid intima-media thickness (cIMT) (-0.03[plusmn]0.01 vs. 0.01[plusmn]0.01 mm; -0.04 [-0.07,-0.01] mm; P= 0.02), logCRP (-0.17[plusmn]0.04 vs. -0.03[plusmn]0.05 mg/L; -0.15 [-0.30,-0.01] mg/L, P=0.04), and triglycerides (-26[plusmn]16 vs. 12[plusmn]8 mg/dL; -37 [-67,-7] mg/dL; P=0.02) improved significantly in the tesamorelin group vs. placebo. In addition, waist circumference (-2[plusmn]1 vs. 1[plusmn]1 cm; -3 [-5,-0.3] cm; P=0.03) and trunk fat decreased (-0.6[plusmn]0.4 vs. 0.7[plusmn]0.4 kg; -1.4 [-2.4,-0.3] kg; P=0.01) and lean mass increased (1.0[plusmn]0.5 vs. -0.4[plusmn]0.4 kg; 1.4 [0.2,2.6] kg; P=0.03). No significant effects on abdominal subcutaneous adipose tissue (SAT) (-6[plusmn]6 vs. 3[plusmn]11 cm[sup]2[/sup]; -10 [-32,13] cm[sup]2[/sup]; P=0.40) or weight were seen. No changes in fasting, 2-hr glucose or HbA1c were seen. Effects of gender were not seen in the model. There were no serious adverse events and no differences in adverse events between the groups.[br][bold]Conclusion:[/bold] Among obese subjects with relative reductions in GH, tesamorelin a GHRH-analogue, selectively reduces VAT, without significant effects on SAT, and improves triglycerides, CRP and cIMT, without aggravating glucose. These data suggest a functional consequence of reduced GH secretion in obesity and demonstrate an improved CVD risk profile resulting from tesamorelin. In addition, this study suggests, more broadly, that strategies to selectively reduce VAT and spare SAT may improve CVD risk in obesity.[br][br]Sources of Research Support: National Institutes of Health Grant K23DK087857 awarded to HM; NIH Grant R01HL085268 awarded to SKG; NIH Grant K24DK064545 awarded to SKG; NIH Grant M01RR01066 and UL1RR025758, Harvard Clinical and Translational Science Center, from the National Center for Research Resources. Partial research funding and study drug was provided by Theratechnologies, Inc. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Center for Research Resources or the National Institutes of Health. This study was registered at www.clinicaltrials.gov as NCT00675506. The study was designed by Dr. Grinspoon. Theratechnologies, Inc. had no input into study design, data analysis, manuscript preparation or decision to publish.[br][br]Disclosures: SKG: Principal Investigator, Theratechnologies; Investigator, Theratechnologies. Nothing to Disclose: HM, MNF, AMR, LCH, MT, HL 2012-06-26T12:30:00 372 2012-06-26T00:00:00 1899-12-30T12:30:00 428 375 2722 OR32-6 OR18-01 Tuesday 2296 2012


2292 ENDO12L_OR33-1 ORAL SESSION: Female Reproduction: Focus on the Ovary (11:15 AM-12:45 PM) Rescue of Obesity-Induced Infertility in Female Mice Due to an Ovarian Theca Cell-Specific Knockout of the Insulin Receptor Sheng Wu, Fredric Wondisford, Chemyong Ko, Po-Ching Lin, Andrew Wolfe Johns Hopkins University School of Medicine, Baltimore, MD; University of Illinois at Urbana-Champaign, Urbana, IL Obesity is associated with many prevalent health problems such as Type 2 DM, polycystic ovary disease (PCOS) and cardiovascular disease, and is linked to reproductive dysfunction. Women with Type 2DM or PCOS present with elevated LH and androgen levels, anovulation and insulin resistance in metabolic tissues. However, previous work from our laboratory revealed that the reproductive axis tissues maintain sensitivity to insulin in the state of obesity. Additionally, fertility is partially protected when the insulin receptor gene (IR) was specifically disrupted in pituitary gonadotrophs of diet induced obese (DIO) mice. The aims of these studies were two-fold; first, to determine whether insulin signaling in the ovary played a role in the development or function of reproductive function, and secondly, to explore whether the ovary is also a target for the pathophysiologic affects of high insulin in obesity. We used conditional knock-out technology to disrupt the IR gene specifically in the theca cells of the ovaries (ThIRKO mouse). No changes in reproductive development or function were observed in lean ThIRKO female mice suggesting that insulin signaling in the theca cell is not essential for reproduction. To explore the effect of insulin signaling in the ovary in obesity we made mice obese (DIO) by feeding them a high fat diet for three months (60% calories from fat). No effect of genotype was observed on fasting glucose or body mass in either lean or DIO mice. As expected, there was a significantly higher fasted glucose and body mass in WT and ThIRKO DIO mice compared to WT and ThIRKO lean mice. DIO WT mice were infertile and experienced increased circulating LH and testosterone levels. By contrast, ThIRKO mice exhibited improved estrous cyclicity and fertility with comparable testosterone level as lean mice. AMH mRNA level of ovary was also expressed differently in WT and ThIRKO. These results confirm that the ovary, like the pituitary, is a target of the high levels of insulin in obesity. These studies highlight an important role for insulin signaling in the ovary in the development of infertility in DIO and indicate that the reproductive axis maintains insulin sensitivity even in the setting of peripheral insulin resistance.[br][br]Nothing to Disclose: SW, FW, CK, P-CL, AW 2012-06-26T11:15:00 342ABDE 2012-06-26T00:00:00 1899-12-30T11:15:00 1357 376 2723 OR33-1 OR200-02 Tuesday 2297 2012


2293 ENDO12L_OR33-2 ORAL SESSION: Female Reproduction: Focus on the Ovary (11:15 AM-12:45 PM) RNA-Binding Protein-Mediated Post-Transcriptional Regulation of Luteinizing Hormone Receptor Expression in the Ovary: Role of miR122 Bindu Menon, Jennifer Sinden, Megan Franzo-Romain, KMJ Menon University of Michigan, Ann Arbor, MI LH receptor mRNA undergoes downregulation during preovulatory LH surge or in response to a pharmacological dose of LH or hCG through a post-transcriptional mechanism that is mediated by an mRNA binding protein, designated as LHR mRNA binding protein (LRBP). The identity of the protein was established as being mevalonate kinase (MVK). miR122, a liver specific microRNA, has been shown to regulate MVK expression. The purpose of this study was to examine whether miR122 is expressed in the ovarian tissue and plays a regulatory role in LHR mRNA expression. Initial experiments were carried out to determine the temporal relationship between miR122 expression and hCG-induced downregulation of LHR mRNA, under [italic]in vivo[/italic] conditions. PMSG-hCG-primed rats were treated with a single dose (50 IU) of hCG to induce LHR mRNA downregulation. Ovaries were collected at different time intervals and miR122 expression was determined by real-time PCR. The results showed that expression of miR122 increased at 30 min and peaked around 1-2h following hCG treatment. Concurrently, mRNA and protein levels of LRBP were higher in hCG-treated ovaries compared to control, reaching a maximum at 2h and 4h, respectively. LHR mRNA levels showed a time-dependent decline after 4h of hCG treatment, reaching a minimum at 12h. The expression of miR122 and its upregulation following hCG treatment was then confirmed using fluorescent [italic]in situ[/italic] hybridization technique. Inhibition of miR122 using an LNA conjugated miR122 specific antagomir, prior to hCG injection, inhibited the hCG-induced increases in LRBP protein levels. Since SREBP-1a, a transcription factor known to regulate the expression of key enzymes in cholesterol biosynthesis, is a possible downstream activator of miR122, the levels of SREBP-1a were examined during hCG-induced downregulation. The results show that SREBP-1a was activated in response to hCG treatment. Taken together, these results suggest that miR122 plays a role in mediating ligand-induced LHR mRNA downregulation in the ovary by regulating the expression of LRBP/MVK. Furthermore, evidence suggests that miR122 regulates LRBP expression through activation of SREBP-1a.[br][br]Sources of Research Support: NIH grant HD06656.[br][br]Nothing to Disclose: BM, JS, MF-R, KMJM 2012-06-26T11:30:00 342ABDE 2012-06-26T00:00:00 1899-12-30T11:30:00 2051 376 2724 OR33-2 OR200-02 Tuesday 2298 2012


2294 ENDO12L_OR33-3 ORAL SESSION: Female Reproduction: Focus on the Ovary (11:15 AM-12:45 PM) IRE1-[alpha] Regulates Cytochrome P450scc Transcription To Mediate Steroidogenesis in the Ovary Rebekah L Viner, Yvonne H Bodenburg, Jie Jiang, Kevin P Rosenblatt, Randall J Urban, Larry Denner University of Texas Medical Branch, Galveston, TX; University of Texas Health Science Center, Houston, TX Polycystic Ovary Syndrome (PCOS) is the most common endocrine disorder affecting women of reproductive age. Cytochrome p450 side chain cleavage enzyme (p450scc) is the rate-limiting enzyme in progesterone biosynthesis and is essential for the normal responses to gonadotropin hormones that maintain fertility and support pregnancy. Therefore, we are interested in identifying factors that regulate the transcription of P450scc. Our laboratory has identified several kinases such as PKCiota, Erk and JNK that are involved in transcriptional activation of P450scc in response to insulin-like growth factor 1 (IGF-1) and the gonadotropin mimetic forskolin. While these kinases mediate cytoplasmic signaling that lead to downstream modulation of P450scc induction, we were interested in identifying kinases that acted more closely to P450scc gene expression. In order to identify novel kinases, we performed a loss-of-function screen using siRNA to all 626 human kinases in a forskolin-stimulated progesterone production high throughput assay in the human KGN granulosa cell line. We found that inositol-requiring enzyme 1 alpha (IRE1) is required for P450scc transcription in response to FSH in the KGN granulosa cell line and to LH in primary granulosa cells (HLGCs) from women undergoing assisted reproduction. Using immunocytochemistry, we have shown that IRE1 is constitutively on the nuclear envelope in large clusters that are typical of its role in the unfolded protein response to ER stress. While this canonical pathway is present and functional in KGN and HLGCs, steroidogenesis does not engage this signaling apparatus. Furthermore, IRE1 knockdown potently down-regulated P450scc mRNA. This data suggests that IRE1 modulates P450scc transcription. Additionally, IRE1 co-immunoprecipitates with ptb-associated splicing factor (PSF) and PSF binding partner, p54nrb/NONO. Our laboratory previously reported that PSF bound the insulin-like growth factor response element (IGFRE) in the P450scc promoter and negatively regulated P450scc transcription. IRE1 may regulate p450scc transcription through a protein interaction complex that includes PSF and p54nrb/NONO. Identifying and understanding the signaling factors that are crucial for normal regulation of P450scc transcription is a major step that will delineate the ovarian dysfunction underlying ovarian-associated fertility disorders such as PCOS.[br][br]Nothing to Disclose: RLV, YHB, JJ, KPR, RJU, LD 2012-06-26T11:45:00 342ABDE 2012-06-26T00:00:00 1899-12-30T11:45:00 1765 376 2725 OR33-3 OR200-02 Tuesday 2299 2012


2295 ENDO12L_OR33-4 ORAL SESSION: Female Reproduction: Focus on the Ovary (11:15 AM-12:45 PM) The Presence of FSH Receptor Polymorphism Asn680Ser Decreases FSH Sensitivity during Ovarian Stimulation for In Vitro Fertilization Aimee Seungdamrong, Peter G McGovern, Andrea Wojtczuk, Laura T Goldsmith New Jersey Medical School - UMDNJ, Newark, NJ Despite years of experience using FSH for ovarian stimulation, the wide variability in ovarian response to FSH, even in younger women with good prognosis for pregnancy success, remains unexplained. We hypothesized that the FSH receptor (FSHR) single nucleotide polymorphism (SNP), which results in a change in amino acid 680 (intracellular domain) from asparagine to serine, affects ovarian response to exogenous FSH administered during controlled ovarian stimulation (COS) for in-vitro fertilization (IVF), manifesting altered ovarian gamete and hormone production.[br]All IVF patients younger than 35 years old were screened for the study between 10/2009 and 12/2011. DNA was extracted from blood samples and genotyping was performed with the ABI Taqman SNP assay (C_2676874_10). Kruskal-Wallis tests and Spearman Rank Correlations were used for data analysis.[br]79 of 194 patients agreed to participate. 4 patients cancelled their IVF cycles for non-medical reasons. The genotype frequency was 0.32 (24/75) for the wild type (Asn/Asn), 0.48 (36/75) for the heterozygote (Asn/Ser), and 0.20 (15/75) for the homozygote SNP (Ser/Ser). Greater exogenous FSH was required per oocyte retrieved for Ser/Ser homozygotes than for heterozygotes (Asn/Ser) or wild types (Asn/Asn) with a significant positive correlation between the serine 680 allele and the amount of exogenous FSH administered per oocyte retrieved (p = 0.002). Furthermore, fewer oocytes were retrieved from Ser/Ser homozygotes than from heterozygotes (Asn/Ser) or wild types (Asn/Asn) with a significant negative correlation between the serine genotype and the total number of oocytes retrieved per patient (p = 0.002). A negative correlation was also found between the serine allele and peak endogenous estradiol levels (p = 0.048). Median patient ages, 32.0 (Asn/Asn), 32.8 (Asn/Ser), and 31.6 (Ser/Ser) years old, were not different (p=0.26). The overall pregnancy rate was 45% (34/75).[br]Presence of the FSH receptor Asn680Ser SNP, in women younger than 35 years old, is correlated with a greater amount of exogenous FSH required for oocyte production and a decreased number of oocytes retrieved after COS for IVF. This FSHR SNP is a potentially clinically important mediator of non-age related variability in endogenous ovarian response to exogenous FSH stimulation. Identification of genetic variation of ovarian response in young, infertile patients may guide their therapy protocols for improved outcomes.[br][br]Sources of Research Support: Ferring Investigator Initiated Grants.[br][br]Nothing to Disclose: AS, PGM, AW, LTG 2012-06-26T12:00:00 342ABDE 2012-06-26T00:00:00 1899-12-30T12:00:00 1823 376 2726 OR33-4 OR200-02 Tuesday 2300 2012


2296 ENDO12L_OR33-5 ORAL SESSION: Female Reproduction: Focus on the Ovary (11:15 AM-12:45 PM) Reduced Oocyte Quality and Poor Embryo Development Due to Dyslipidemia or Obesity Is Normalized by Blocking Endoplasmic Reticulum Stress Linda L Wu, Darryl L Russell, Robert J Norman, Rebecca L Robker University of Adelaide, Adelaide, Australia Obese women often experience infertility due to anovulation, decreased pregnancy rates and increased miscarriages. Our previous studies using a mouse model showed that female mice with diet-induced obesity are also less likely to ovulate and, when fertilized, their oocytes exhibit abnormal embryo development (1). These defects in ovulation and oocyte quality were associated with endoplasmic reticulum (ER) stress responses in cumulus-oocyte complexes (COCs), intracellular lipid accumulation and altered mitochondrial activity in oocytes (2). The current studies sought to determine whether lipid-induced ER stress directly alters oocyte mitochondrial activity and thereby leads to impaired early embryo growth.[br]Treatment of mouse COCs with high physiological levels of the fatty acid palmitic acid significantly induced ER stress marker genes ([italic]Atf4[/italic], [italic]Atf6[/italic], [italic]Xbp1s[/italic], [italic]Hspa5[/italic]) and IRE1[alpha] phosphorylation, demonstrating an ER stress response that was reversible with an ER stress inhibitor. Pentraxin-3, an extracellular matrix protein essential for fertilization, showed dramatically impaired secretion concurrent with ER stress. Oocytes matured in high dose palmitic acid had significantly reduced mitochondrial activity, reduced in vitro fertilization rates and delayed blastocyst development. The deficiencies in protein secretion, mitochondrial activity and oocyte developmental competence were each normalized by ER stress inhibitor, demonstrating that ER stress is a key mechanism mediating fatty acid-induced defects in oocyte mitochondrial activity.[br]To determine whether ER stress inhibitors can reverse obesity-induced female infertility in vivo we utilized a novel mouse, the [apos]Blobby[apos] mutant mouse which exhibits hyperphagia and extreme obesity even on a chow diet. Obese Blobby mice had reduced ovulation rates compared to age-matched wildtype and heterozygous littermates and the oocytes that did ovulate showed significantly reduced mitochondrial activity and poor embryo development when fertilized in vitro. Remarkably ovulation rates, oocyte mitochondrial activity and blastocyst development in the obese Blobby mice were restored to normal when they were treated with once daily injection of ER stress inhibitor for just 4 days. These results clearly demonstrate that ER stress is a fundamental mechanism involved in the female infertility and poor embryo growth that is associated with obesity but that it can be reversed in order to restore blastocyst development to normal.[br][br](1) Minge CE et al., Endocrinology 2008;149:2646. (2) Wu LL et al., Endocrinology 2010; 151:5438.[br][br]Sources of Research Support: National Health and Medical Research Council of Australia; Channel 7 Children[apos]s Research Foundation; Women[apos]s and Children[apos]s Hospital Foundation.[br][br]Nothing to Disclose: LLW, DLR, RJN, RLR 2012-06-26T12:15:00 342ABDE 2012-06-26T00:00:00 1899-12-30T12:15:00 1498 376 2727 OR33-5 OR200-02 Tuesday 2301 2012


2297 ENDO12L_OR33-6 ORAL SESSION: Female Reproduction: Focus on the Ovary (11:15 AM-12:45 PM) [italic]In Vitro[/italic] Ovulation and Luteinization of Murine, Rhesus, and Human Follicles Robin M Skory, Teresa K Woodruff Northwestern University, Chicago, IL Ovulation is a dynamic process thought to involve several ovarian microenvironments leading to the release of a mature egg. During luteinization, the somatic cells differentiate to become the corpus luteum, which produces hormones to maintain reproductive cyclicity or pregnancy. The present study asks whether these two processes are autonomous to the follicle or depend on other cells or structures in the local environment. Three-dimensional culture systems have been developed to mature follicles in vitro and alginate has proven to be a robust supportive biomaterial for all follicular phase activities. To investigate mechanisms of follicular rupture, antral murine follicles were isolated, encapsulated and cultured in alginate. When follicles reached diameters greater than 285[mu]m, hCG was added and follicles were removed from alginate. Sixteen hours post-hCG, 89% of follicles ruptured (n=38). Of the follicles that ruptured, 91% contained meiotically competent (MII) oocytes. To assess luteinization of the somatic cell compartment, hCG was added to follicles cultured in alginate and culture was continued for an additional 6 days. Post-hCG treatment, follicles demonstrated characteristics of structural and functional luteinization. Namely, estradiol synthesis decreased 10 fold and progesterone synthesis increased 80 fold. In addition, rodent follicles luteinized in vitro showed decreasing levels of inhibin A and B as shown by EIA and Western Blot, mimicking in vivo patterns. To examine luteinization of higher mammalian species, follicles from rhesus and human were similarly cultured in alginate. Post-hCG administration, follicle cells luteinized and increased secretion of progesterone. Importantly, inhibin A expression also increased, which accurately phenocopies the species difference between rodent and primate. These results show that follicular rupture is a follicle-autonomous process, occurring independent of the reticuloendothelial system or ovarian surface epithelium. In contrast to 2D cultures, follicles cultured in the 3D alginate system accurately mimicked in vivo hormone patterns. By harnessing the ability to examine several ovarian microenvironments in vitro, we can identify regulatory mechanisms underlying the multi-step process of ovulation. In the future this system may be applied to examine hormonal regulation in the peri-ovulatory period and to screen novel compounds for contraception.[br][br]Sources of Research Support: NIH/NICHD U54HD041857 and NIH/NIA F30AG040916.[br][br]Nothing to Disclose: RMS, TKW 2012-06-26T12:30:00 342ABDE 2012-06-26T00:00:00 1899-12-30T12:30:00 2351 376 2728 OR33-6 OR200-02 Tuesday 2302 2012


2298 ENDO12L_OR34-1 ORAL SESSION: Glucocorticoid Actions (11:15 AM-12:45 PM) [italic]Bcl[/italic]I Glucocorticoid Receptor Polymorphism Is Associated with Greater Body Fatness and Higher Insulin Resistance: The Hoorn and CODAM Studies CCM Geelen, MMJ van Greevenbroek, NC Schaper, G Nijpels, LM [apos]t Hart, CG Schalkwijk, I Ferreira, CJH van der Kallen, EFC van Rossum, HP Sauerwein, JM Dekker, CDA Stehouwer, B Havekes Maastricht University Medical Center, Maastricht, Netherlands; Maastricht University Medical Center, Maastricht, Netherlands; VU University Medical Center, Amsterdam, Netherlands; Leiden University Medical Center, Leiden, Netherlands; Leiden University Medical Center, Leiden, Netherlands; Maastricht University Medical Center, Maastricht, Netherlands; Erasmus Medical Center, Rotterdam, Netherlands; Maastricht University Medical Center, Maastricht, Netherlands [bold]Background[/bold]: Glucocorticoid receptor (GR) polymorphisms play an important role in the downstream effects of glucocorticoids (GCs). The [italic]Bcl[/italic]I polymorphism has been shown to be associated with an enhanced GC sensitivity, which may result in greater abdominal obesity and its metabolic consequences. We have investigated the association of the [italic]Bcl[/italic]I polymorphism with body fatness/fat distribution and insulin resistance in a large sample combining data from 2 cohort studies, which were both enriched with participants with a disturbed glucose metabolism and type 2 diabetes (DM2).[br][bold]Patients and methods[/bold]: The present study included 1228 participants (aged 64.7 [plusmn] 8.5 years, 22.6% had impaired glucose metabolism and 32.6% had DM2) from the Cohort study on Diabetes and Atherosclerosis Maastricht (CODAM, n=543) and the Hoorn Study (n=685), who were genotyped for the [italic]Bcl[/italic]I polymorphism using Taqman SNP genotyping assays; 519 (42%) were non-carriers (CC), 540 (44%) were heterozygous (CG) and 169 (14%) were homozygous (GG) carriers of the G-allele. Linear regression analyses adjusted for cohort, sex, age, glucose metabolism, smoking and use of medication were used to compare carriers of one or two G-alleles to non-carriers (dominant model; CG+GG vs. CC) and homozygous carriers to carriers of one or no G-allele (recessive model; GG vs. CC+CG). BMI (kg/m2), waist and hip circumferences (cm), and waist-hip-ratio (WHR) were used as measures of total fat and fat distribution. Insulin resistance was estimated using the standardized homeostatic model assessment of insulin resistance index (HOMA2-IR).[br][bold]Results[/bold]: In the fully adjusted recessive model, individuals with the GG genotype had a higher BMI (regression coefficient [beta]=1.05 (95% confidence interval: 0.41-1.70), p=0.001), waist ([beta]=2.66 (0.90-4.41), p=0.003) and hip circumference ([beta]=2.25 (0.86-3.64), p=0.002), and a higher HOMA2-IR ([beta]=0.08 (0.01-0.16), p=0.034) when compared to the CC+CG genotype. There was no difference in WHR. In the dominant model, waist and hip circumferences tended to be higher in CG+GG carriers than in non-carriers of the G-allele ([beta]=1.07 (-0.16-2.30), p=0.087 and [beta]=0.79 (-0.18-1.77), p=0.110 resp.). Genotypes were consistent with Hardy-Weinberg equilibrium (p[gt]0.05) and the observed effects were largely comparable in both cohorts.[br][bold]Conclusion[/bold]: This study demonstrates that homozygous G-allele carriers of the [italic]Bcl[/italic]I polymorphism have a significantly greater total body fatness and higher insulin resistance.[br][br]Nothing to Disclose: CCMG, MMJvG, NCS, GN, LMtH, CGS, IF, CJHvdK, EFCvR, HPS, JMD, CDAS, BH 2012-06-26T11:15:00 361 2012-06-26T00:00:00 1899-12-30T11:15:00 671 377 2729 OR34-1 OR02-01 Tuesday 2303 2012


2299 ENDO12L_OR34-2 ORAL SESSION: Glucocorticoid Actions (11:15 AM-12:45 PM) Glucocorticoids Increase Subcutaneous Adipose Tissue Insulin Sensitivity [italic]In Vivo[/italic]: A Randomized Double-Blind, Placebo Controlled, Cross-Over Study Jonathan Hazlehurst, Matthew Armstrong, Sarah Borrows, Laura L Gathercole, Paul M Stewart, Jeremy W Tomlinson University of Birmingham, Birmingham, UK; University of Birmingham, Birmingham, UK; Queen Elizabeth Hospital, Birmingham, UK Glucocorticoid (GC) excess, Cushing[apos]s syndrome, is characterized by central obesity, proximal myopathy, insulin resistance and in some cases type 2 diabetes mellitus. Current dogma suggests that GCs cause insulin resistance in all tissues. We have previously demonstrated that GCs cause insulin sensitization of human adipose tissue[italic] in vitro,[/italic] whilst inducing insulin resistance in human skeletal muscle. No prior study has evaluated whether these observations translate into the clinical setting.[br]15 healthy volunteers (age 33[plusmn]1.0 years, BMI 26.6[plusmn]1.0kg/m[sup]2[/sup], fasting glucose 4.7[plusmn]0.1mmol/L) were recruited into a double-blind, randomized, placebo controlled, cross-over study, in which they received both an overnight saline and hydrocortisone (HC) infusion (0.2mg/kg/h) in a random order. Volunteers underwent a 2-step hyperinsulinaemic (low dose and high dose) euglycaemic clamp. In addition, adipose tissue microdialysis was performed to determine insulin-stimulated pyruvate generation and insulin-mediated suppression of glycerol release (lipolysis).[br]HC infusion elevated fasting glucose levels (4.4[plusmn]0.1 vs. 6.0[plusmn]0.1mmol/L, p[lt]0.0001) and caused systemic insulin resistance as measured by HOMA-IR (1.1[plusmn]0.2 vs. 2.0[plusmn]0.4, p[lt]0.05) and decreased glucose infusion rates (M-value) following high dose insulin (9.1[plusmn]0.5 vs. 7.4[plusmn]0.7mg/kg/min, p[lt]0.01). In adipose tissue, in the fasting state, pyruvate release was similar between HC and saline. As expected, insulin stimulated pyruvate release (99[plusmn]13 vs. 136[plusmn]20[micro]mol/L/h, p[lt]0.05) but this was dramatically enhanced following overnight HC infusion consistent with insulin sensitization (136[plusmn]20 vs. 281[plusmn]27[micro]mol/L/h, p[lt]0.005). Insulin suppressed adipose tissue lipolysis (235[plusmn]47 vs. 53[plusmn]15[micro]mol/L/h, p[lt]0.00001) whilst in contrast, HC infusion increased rates of lipolysis (235[plusmn]47 vs. 365[plusmn]53[micro]mol/L/h, p=0.05). However, the insulin-mediated suppression of lipolysis was significantly enhanced following overnight HC infusion (reduction in lipolytic rate; -114[plusmn]25 vs. -196[plusmn]39[micro]mol/L/h, p=0.05), again, consistent with adipose tissue insulin sensitization.[br]This study represents the first description of adipose tissue insulin sensitization by GC [italic]in vivo,[/italic] and clearly demonstrates tissue specific actions of GCs to interact with, and modify insulin sensitivity and action. It defines an important advance in our understanding of the actions of both endogenous and exogenous GCs and may have implications for the development and targeting of future GC therapies.[br][br]Sources of Research Support: Medical Research Council Senior Clinical Fellowship (ref. G0802765) awarded to JWT.[br][br]Nothing to Disclose: JH, MA, SB, LLG, PMS, JWT 2012-06-26T11:30:00 361 2012-06-26T00:00:00 1899-12-30T11:30:00 797 377 2730 OR34-2 OR02-01 Tuesday 2304 2012


2300 ENDO12L_OR34-3 ORAL SESSION: Glucocorticoid Actions (11:15 AM-12:45 PM) Glucocorticoid Dose in Adults with CAH; Association with Clinical and Biochemical Variables [mdash] Analysis of the United Kingdom Congenital Adrenal Hyperplasia Adult Study Executive (CaHASE) Cohort Thang S Han, Stimson H Roland, Rees Aled, Krone Nils, Willis S Debbie, Wild H Sarah, Conway S Gerard, Arlt Wiebke, Brian R Walker, Ross J Richard, and CaHASE University College London, London, UK; University of Edinburgh, Edinburgh, UK; Cardiff University, Cardiff, UK; University of Birmingham, Birmingham, UK; Society for Endocrinology, Bristol, UK; University of Edinburgh, Edinburgh, UK; University of Sheffield, Sheffield, UK [bold]Objectives: [/bold]We have previously reported that metabolic abnormalities are common in adults with CAH. We hypothesised that type of glucocorticoid treatment and dose mediates the adverse metabolic outcome.[br][bold]Design: [/bold]Patients (n=196) were treated with hydrocortisone (n=25M,26W), prednisolone (n=21M,67W), dexamethasone (n=15M,22W) or combination therapy (n=4M,16W). Treatment dose was converted to prednisolone equivalent using respective ratios of 1:5:0.15 for prednisolone: hydrocortisone: dexamethasone. Data were examined by ANOVA, univariate regression and the following variables were selected for principal components analysis (PCA): 6 metabolic measures (waist, triglycerides, HDL-cholesterol, HOMA-IR, systolic (SBP) and diastolic (DBP) blood pressure), 3 disease control factors (androstenedione, 17-hydroxyprogesterone (OHP) and testosterone) and mutation severity. The validity of analysis was confirmed by Kaiser-Meyer-Olkin measure of sampling adequacy (0.66) and Bartlett[apos]s test of sphericity (208, P[lt]0.001).[br][bold]Results: [/bold]ANOVA and univariate regression analysis showed weak positive correlations ([italic]r[/italic][lt]0.2) between prednisolone equivalent dose and SBP, DBP, HDL-cholesterol and HOMA-IR. For PCA, 3 principal components (PCs) with eigenvalues [gt]1 were retained, explaining 62% of the total variance in the observed variables. 4 items (waist, triglycerides, HOMA-IR and HDL (negative loading)) were found to load on PC1, described as [ldquo][italic]adiposity and insulin resistance[/italic][rdquo], 3 items (androstenedione, 17-OHP and testosterone) on PC2, described as [ldquo][italic]disease control[/italic][rdquo], and 3 items (SBP, DBP and mutation severity) on PC3, described as [ldquo][italic]blood pressure and mutations[/italic][rdquo]. The resultant factor scores were analysed by stepwise multiple regression analysis (age and sex adjusted) to assess their relationship with prednisolone equivalent dose. PC2 (b=0.62, 95%CI: 0.10 to 1.14, P=0.020) and PC3 (b=1.05, 95%CI: 0.53 to 1.57, P[lt]0.001) were retained accounting for 23% of the explained variance. When glucocorticoid type was included, only PC3 (b=0.85, 95%CI: 0.40 to 1.31, P[lt]0.001) and glucocorticoid type (b=-1.07, 95%CI: -1.45 to -0.70, P[lt]0.001) were retained explaining 43% of the total variance.[br][bold]Conclusions: [/bold]Increasing total glucocorticoid dose to treat CAH is positively associated with disease control, BP and mutation severity and negatively with the use of dexamethasone, but is not associated with adiposity or insulin resistance in this cohort.[br][br](1) Arlt et al JCEM 2010;95:5110-21.[br][br]Sources of Research Support: CaHASE are greatful to the Clinical Endocrinology Trust for financial support and the Society for Endocrinology for management of he project.[br][br]Disclosures: RJR: Speaker, Diurnal. Nothing to Disclose: TSH, SHR, RA, KN, WSD, WHS, CSG, AW, BRW 2012-06-26T11:45:00 361 2012-06-26T00:00:00 1899-12-30T11:45:00 733 377 2731 OR34-3 OR02-01 Tuesday 2305 2012


2301 ENDO12L_OR34-4 ORAL SESSION: Glucocorticoid Actions (11:15 AM-12:45 PM) Genetic Deletion of Uterine Glucocorticoid Receptor Leads to Profound Defects in Fertility Shannon D Whirledge, Francesco J DeMayo, John P Lydon, John A Cidlowski National Institute of Environmental Health Sciences, Research Triangle Park, NC; Baylor College of Medicine, Houston, TX Glucocorticoids regulate a broad spectrum of biological processes critical for life, including those involved in immune and inflammatory reactions as well as others with implications for fertility. In response to stress, including infection, malnutrition, anxiety and depression, glucocorticoids secreted from the adrenal glands can suppress reproductive functions along the hypothalamo-pituitary-gonadal (HPG) axis. Glucocorticoids exert their effects through binding the glucocorticoid receptor (GR), a member of the nuclear steroid receptor superfamily that functions as a ligand-dependent transcription factor to regulate the expression of glucocorticoid-responsive genes. Localization of GR to specific cell types within the HPG axis suggests a direct influence on reproductive function at the gonadal level. To clearly define those physiological events specifically attributable to uterine expressed GR in vivo, we have generated a mouse model that enables conditional ablation of GR function specifically in the uterus. Exons 3 and 4 of the murine GR gene were floxed to generate a conditional GR[sup]fl[/sup] allele. Crossing the GR[sup]fl[/sup] mice with progesterone receptor (PR)-Cre transgenic mice results in Cre excision restriced to cells that express PR, such as in the uterus. Male and female mice homozygous for the GR mutation develop normally to adulthood. However, the adult female GR mutant display profound defects in fertility. These include a delay to pregnancy (WT 22.2 [plusmn] 1.5 days v. KO 97 [plusmn] 55.6 days) and smaller number of litters per reproductive testing cycle (WT 4 [plusmn] 1.8 total litters v. KO 2 [plusmn] 2.16 total litters). Further characterization of the GR mutant mouse reveals altered expression signaling in the innate and adaptive immune response, an important aspect of implantation and survival of the semiallogeneic fetus. These results suggest GR plays a role in fertility specific to the uterus.[br][br]Sources of Research Support: This research was supported by the Intramural Research Program of the NIH, National Institute of Environmental Health Sciences.[br][br]Nothing to Disclose: SDW, FJD, JPL, JAC 2012-06-26T12:00:00 361 2012-06-26T00:00:00 1899-12-30T12:00:00 1588 377 2732 OR34-4 OR02-01 Tuesday 2306 2012


2302 ENDO12L_OR34-5 ORAL SESSION: Glucocorticoid Actions (11:15 AM-12:45 PM) 11[beta]-Hydroxysteroid Dehydrogenase Type 1 Mediates Age- and Glucocorticoid-Induced Skin Thinning: Reversal of the Aging Phenotype in 11[beta]-HSD1 Null Mice Ana Tiganescu, Andrew E Mayes, Elizabeth H Rabbitt, Gareth G Lavery, Mark S Cooper, Paul M Stewart IBR, University of Birmingham, Birmingham, UK; Colworth Science Park, Sharnbrook, Bedford, UK [bold]Background:[/bold] Glucocorticoid (GC) excess adversely affects skin integrity, inducing thinning and impaired wound healing. Aged skin shows features similar to GC therapy with changes most marked in photo-exposed (PE) skin. We have demonstrated expression and function of the GC-activating (cortisone to cortisol) enzyme 11b-hydroxysteroid dehydrogenase type 1 (11b-HSD1) in skin and human dermal fibroblasts (HDF).[br][bold]Methods:[/bold] 11b-HSD1 expression in mouse and human skin and HDF from photo-protected (PP) and PE sites of varying donor age. Microfluidic card real-time PCR to analyse mRNA levels for 108 genes involved in collagen biosynthesis, signalling and stress responses +/- an 11b-HSD1-specific inhibitor. The skin phenotype of aged 11b-HSD1-null (KO) mice was investigated by histological, dermal cellularity and collagen density (Masson Trichrome).[br][bold]Results:[/bold] 11b-HSD1 activity increased by 42%* in aged human biopsies ([gt]60 vs. 20-30 years, n=20) and similarly by 69%* in aged mouse skin ([gt]90 vs. [lt]20 weeks, n=5). 11[beta]-HSD1 activity was elevated in PE vs. PP samples from young and aged human donors by 34%*** and 21%* respectively. Donor-matched PE HDF exhibited 6.5-fold*** increased 11b-HSD1 mRNA relative to PP samples (n=10). 23 age-related 11b-HSD1-regulated genes were identified; dual specificity phosphatase-1 (MAPK p38 inhibitor) increased in aged PP/PE biopsies (1.6-fold*/2.3-fold* respectively) and was decreased with inhibitor (0.7-fold*). Decreased p38 signalling impairs macrophage function and healing responses. Age-induced skin atrophy in WT mice was prevented in 11[beta]-HSD1 KO mice. Dermal cellularity which decreased by 40%* in aged WT was normalized in KO mice and collagen density was preserved.[br][bold]Conclusion:[/bold] 11b-HSD1 expression is increased in ageing and PE skin. 11[beta]-HSD1 inhibition and 11[beta]-HSD1 deletion in mice prevented the development of age-related changes in skin by preserving fibroblast integrity and collagen content. * = p[lt]0.05, ** = p[lt]0.01, *** = p[lt]0.001.[br][br]Nothing to Disclose: AT, AEM, EHR, GGL, MSC, PMS 2012-06-26T12:15:00 361 2012-06-26T00:00:00 1899-12-30T12:15:00 2094 377 2733 OR34-5 OR02-01 Tuesday 2307 2012


2303 ENDO12L_OR34-6 ORAL SESSION: Glucocorticoid Actions (11:15 AM-12:45 PM) Immunoglobulin Class Switch Recombination Is Acutely Regulated by Glucocorticoids in Human B Lymphocytes Ann M Benko, Nancy J Olsen, William J Kovacs Pennsylvania State University, College of Medicine, Hershey, PA; Pennsylvania State University, College of Medicine, Hershey, PA B lymphocytes, when activated by antigen in the presence of a costimulatory signal, initiate processes by which the class of immunoglobulin produced by the cell is changed from IgM to IgG, IgE, or IgA. This is accomplished by genetic recombination events at the immunoglobulin heavy chain constant region (C[sub]H[/sub]) gene, and is initiated by activation-induced cytidine deaminase (AID), an enzyme that deaminates cytosines in switch regions of the C[sub]H[/sub] locus. Subsequent enzymatic modification of the DNA creates double-strand breaks with DNA ends that are capable of end-joining recombination. This class switch recombination (CSR) event results in an excised circular DNA fragment (switch circle) that remains transcriptionally active--and quantitation of such switch circle transcripts permits detection of ongoing CSR activity. We examined the effects of physiologic concentrations of glucocorticoids on CSR by measuring changes in expression of both AID mRNA and switch circle transcripts in activated B cells. We used real-time polymerase chain reaction (PCR) for quantitation of AID mRNA levels by the [Delta][Delta]Ct method and established an assay for IgM to IgG1 switch circle transcripts using a cloned standard template. Purified human B cells were prepared by positive selection using CD19-coated magnetic beads and were activated [italic]in vitro[/italic] by treatment for 18-20 hours with IL-4 and anti-CD40, resulting in induction of AID mRNA expression by 13.53 [plusmn] 2.98-fold. Treatment with dexamethasone (Dex) resulted in a dose-dependent inhibition of AID mRNA expression with half-maximal effect observed at concentrations under 10 nM. Dex at 10nM concentration reduced AID mRNA levels by an average of 55.72% (p = 0.0009). Dose dependent reduction in switch circle transcript expression by Dex treatment was also observed (30.45 % at 10nM; p = 0.0045). The observed effects were reversed by co-incubation with the GR antagonist mifepristone. Cell viability was unaltered by glucocorticoid treatment ([gt]93% live cells in all treatments). Activation of B cells was accompanied by up-regulation (14.71 [plusmn] 3.21-fold) of mRNA encoding 11[beta] HSD1, a glucocorticoid-regenerating enzyme. These data demonstrate a previously unrecognized immunomodulatory property of glucocorticoids--to inhibit immunoglobulin class switch recombination in activated B lymphocytes. The observed increase in expression of 11[beta] HSD1 during B cell activation suggests a potential role for endogenous glucocorticoid regeneration in modulating CSR.[br][br]Nothing to Disclose: AMB, NJO, WJK 2012-06-26T12:30:00 361 2012-06-26T00:00:00 1899-12-30T12:30:00 312 377 2734 OR34-6 OR02-01 Tuesday 2308 2012


2304 ENDO12L_OR35-1 ORAL SESSION: Neoplasia of Endocrine Tissues: Observations That Are Likely To Impact Clinical Management (11:15 AM-12:45 PM) A Murine Model Supports the Role of PRKAR1A as a Tumor Suppressor Gene in the Pancreas Emmanouil Saloustros, Sisi Liu, Woo-Jin Song, Matthew Starost, Elizabeth Geras-Raaka, Mehboob Hussain, Constantine Stratakis National Institute of Child Health and Human Development, Bethesda, MD; Johns Hopkins University, Baltimore, MD; National Institutes of Health, Bethesda, MD; National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD Background: Carney complex (CNC) is a rare disease inherited as an autosomal dominant trait, associated with various tumors, and caused most frequently by inactivation of the PRKAR1A gene which codes for protein kinase A (PKA), cAMP-dependent, regulatory, type I, a-subunit. Recently a possible association of pancreatic neoplasms with CNC was reported in nine out of 354 patients (2.5%), suggesting the potential role of PRKAR1A as a tumor suppressor gene in the pancreatic tissue. A genetically engineered mouse model with tissue specific prkar1a ablation is likely to elucidate valuable insights on the role of prkar1a as a tumor suppressor gene in the pancreas.[br]Methods: Prkar1afl/fl and Pdx-1-CRE mice were previously reported (1,2) and were kept in a mixed genetic background (C57Bl/6 x FVB). Pdx1 is a transcription factor expressed in beta-insulin producing cells and in some a-glucagon producing cells of the mouse adult pancreatic islet. Both prkar1a alleles were conditionally deleted during development by generating Pdx1-Cre; prkar1af/f mice ([Delta][ndash]prkar1a). Mice were phenotyped at the age of two, four and six months and tissues were collected for histological and molecular analysis.[br]Results: Mice developed invasive carcinomas with 100% penetrance by the age of 4-5 months. First appearance of the lesions was as early as at the age of 8-10 weeks of age. Histologically the lesions resembled normal-looking islet cells arranged in cords or trabeculae of cells with bland round to oval nuclei with small nucleoli and modest amounts of cytoplasm. Malignancy defined by stromal invasion. Immunohistochemistry revealed decreased staining for prkar1a compared to the normal islets, while immonofluorescence-using probes for insulin, glucagon and c-peptide confirmed an endocrine component for these lesions. As expected based on pdx1 expression, a mixed cell population of beta (mostly) and alpha cells composed the tumors. Since overnight fasting (18h) did not reveal any differences in glucose levels between [Delta][ndash]prkar1a and littermates with normal or haploinsufficient prkar1a allele, the probability that these tumors are functional is low, although ongoing work aims at characterizing them better.[br]Conclusions: Loss of prkar1a function predisposes pancreas to invasive carcinomas. This supports the role of prkar1a as a tumor suppressor gene in the pancreas and provides valuable tools to evaluate novel therapeutics in pancreatic malignancies.[br][br](1)Kirschner et al., Cancer Res. 2005;65:4506. (2)Lammert et al., Science 2001;294:564.[br][br]Nothing to Disclose: ES, SL, W-JS, MS, EG-R, MH, CS 2012-06-26T11:15:00 352DEF 2012-06-26T00:00:00 1899-12-30T11:15:00 2228 378 2735 OR35-1 OR44-01 Tuesday 2309 2012


2305 ENDO12L_OR35-2 ORAL SESSION: Neoplasia of Endocrine Tissues: Observations That Are Likely To Impact Clinical Management (11:15 AM-12:45 PM) Novel Therapeutic Approaches to the Treatment of Estrogen Receptor-Negative Breast Cancer Guowei Gu, Kyle Covington, Yassine Rechoum, Bert O[apos]Malley, David Mangelsdorf, John Minna, Paul Webb, Suzanne Fuqua Baylor College of Medicine, Houston, TX; The University of Texas, Dallas, TX; The Methodist Hospital Research Institute, Houston, TX [bold]Background: [/bold]The treatment of estrogen receptor (ER)-negative breast cancer (BC) is a major clinical problem due to the lack of useful therapeutic targets. Nuclear receptors (NRs) are potential targets in these patients because they regulate global transcriptional events and many already have agonists/antagonists available.[br][bold]Material and Methods: [/bold]We used microarray analysis of 227 ER-negative tumors, and performed hierarchical clustering using 41 NRs. Receptors were scored using prediction analysis of microarrays (PAM) across clustered groups. Cell lines were matched to subtypes using published data (1). pGIPZ lentiviral vectors were used to knockdown candidates. MTT and soft agar assays were used to measure chemosensitivity.[br][bold]Results: [/bold]The 41 NRs clustered tumors into 5 groups. For each group we selected genes representing the highest ranked discriminators. Thyroid hormone receptor [beta] (THR[beta]) and COUP-TF1 were selected from group V. The expression levels of the selected receptors were confirmed by qRT-PCR and Western blot.[br]Knockdown of THR[beta] in ER-negative HCC2185 cells rendered cells more resistant to all chemotherapeutics by using MTT. Similar results were confirmed in ER-negative MDA-MB-453 and HCC202 cells. Knockdown of THR[beta] enhanced colony forming potential in anchorage-independent soft agar assays in MDA-MB-453 and HCC202 cells. COUP-TFI was found to be upregulated in HCC2185 and MDA-MB-453 cells with THR[beta] knockdown. Statistical analysis using clinical data from Sabatier et al. (2) showed that patients with low THR[beta] or high COUP-TFI have a worse clinical outcome. In order to translate these findings into the clinic, we treated cells with a specific THR[beta] agonist, GC-1. GC-1 inhibited cell growth, and synergistic effects were observed when cells were treated with GC-1 and Docetaxel in combination. Re-expression of ER[alpha] protein was observed in ER-nagative cells lines concommittant with THR[beta] knockdown, suggesting that modulation of THR[beta] may also extend hormonal therapy to this hormonally insensitive group of patients.[br][bold]Conclusion: [/bold]Clinical targeting of NRs in ER-negative BCs is a novel strategy since receptors can be specifically targeted with ligands. Our data suggest that chemotherapy response in ER-negative patients overexpressing THR[beta] could be enhanced with a THR[beta] agonist. Similarly, functional re-activation of ER[alpha] might extend hormonal therapies to these patients as well.[br][br](1)Neve et al., Cancer Cell. 2006 Dec;10(6):515-27. (2)Sabatier et al., Mol Cancer. 2011 Jul 21;10:86.[br][br]Sources of Research Support: CPRIT RPI 100188 and. Komen for the Cure KG0816494.[br][br]Nothing to Disclose: GG, KC, YR, BO, DM, JM, PW, SF 2012-06-26T11:30:00 352DEF 2012-06-26T00:00:00 1899-12-30T11:30:00 1774 378 2736 OR35-2 OR44-01 Tuesday 2310 2012


2306 ENDO12L_OR35-3 ORAL SESSION: Neoplasia of Endocrine Tissues: Observations That Are Likely To Impact Clinical Management (11:15 AM-12:45 PM) Impact of Tamoxifen in the Chemoprevention of Bladder Cancer Suraj Konnath George, Steven S Shen, Patricia M Dillard, Seth P Lerner, Carolyn L Smith Baylor College of Medicine, Houston, TX; The Methodist Hospital Research Institute, Houston, TX; Baylor College of Medicine, Houston, TX Transitional cell carcinoma of the bladder is three times more common in men than in women with [sim]74,000 new cases anticipated for 2012 in the US. Most bladder carcinomas are a low grade papillary phenotype and frequently reoccur. Therapeutics able to prevent recurrence are a significant unmet need. Estrogen receptors (ERs) are ligand-regulated transcription factors which promote cellular growth and survival, and treatment with the selective estrogen receptor modulator (SERM) tamoxifen, can block the growth of estrogen-dependent cells. ER[beta] is the dominant ER expressed in bladder cancer cell lines and human bladder tumors. The goal of this study was to determine the impact of tamoxifen on bladder carcinogenesis induced by N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) supplied [italic] ad libitum [/italic] in drinking water (0.05%) for 12 weeks to C57BL/6 male mice. Tamoxifen ([sim]16, 28 and 55 [mu]g/day) was administered via slow-release pellets concurrent with BBN treatment when animals were 8-20 wks old (n=21-25/group). The study was terminated at 27 weeks. The mean bladder weights obtained from BBN only animals were significantly higher than control animals that did not receive BBN. This corresponded to a 100% incidence of urothelial cancer in the BBN only mice of which 40% developed muscle-invasive tumors; controls were histologically normal. Treatment of mice with tamoxifen resulted in bladder weights that were statistically different from the BBN only group. Histopathological analyses for the three tamoxifen-treated groups revealed normal histology in 54%, 59% and 46% of the 16, 28 and 55 [mu]g/day tamoxifen groups, respectively. Proliferation, assessed by Ki67 staining, was significantly elevated for the BBN only group in comparison to the control as well as the BBN+tamoxifen groups. Immunohistochemistry revealed ER[beta] expression in all groups. Unexpectedly, ER[alpha] was detected in the bladders of animals exposed to BBN, but not found in any control mice. However, levels of ER[alpha] mRNA were similar for control, BBN and BBN+tamoxifen groups, indicating that the ER[alpha] detected by immunohistochemistry in all BBN-treated groups arises from a BBN-induced post-transcriptional event. These data indicate that tamoxifen administered concurrently with BBN prevents bladder carcinogenesis, and that this protective effect may be mediated by ER[alpha] and/or ER[beta]. Administration of tamoxifen may be a therapeutic option for secondary prevention of human bladder cancer.[br][br]Nothing to Disclose: SKG, SSS, PMD, SPL, CLS 2012-06-26T11:45:00 352DEF 2012-06-26T00:00:00 1899-12-30T11:45:00 1846 378 2737 OR35-3 OR44-01 Tuesday 2311 2012


2307 ENDO12L_OR35-4 ORAL SESSION: Neoplasia of Endocrine Tissues: Observations That Are Likely To Impact Clinical Management (11:15 AM-12:45 PM) The Cholesterol Recognition Amino Acid Consensus (CRAC) Motif of the 18-kDa Translocator Protein (TSPO) as a Chemotherapeutic Target Andrew Midzak, Rohan Pirangute, Vassilios Papadopoulos The Research Institute of the McGill University Health Centre, Montr[eacute]al, Canada; McGill University, Montr[eacute]al, Canada Increased expression of the 18-kDa translocator protein (TSPO) gene has been described in a variety of cancers, including prostate, ovarian and breast carcinomas. Moreover, the expression of TSPO correlates with the metastatic potential and invasiveness of tumors, making it an important diagnostic and potentially therapeutic target for oncology. TSPO is a mitochondrial high-affinity cholesterol and drug-binding protein best known for its critical role in steroid production, and though its drug-binding site has been of significant research interest, the TSPO cholesterol binding site has received comparatively less research attention. This cholesterol binding site of TSPO is located at the cytosolic C-terminus of the protein and is constituted by a phylogenetically conserved cholesterol recognition/interaction amino acid consensus (CRAC) motif. As part of our study of TSPO, we have previously computationally modeled and biologically screened a library of compounds targeted against the CRAC domain (1). These bioassays focused on the ability to nontoxically inhibit steroid production, but a series of compounds were identified that exhibited cytotoxicity, leading us to hypothesize that targeting the CRAC motif with novel ligands could alter TSPO function and in turn cancer cell proliferation and viability. The TSPO CRAC motif was targeted with novel synthetic drugs either alone or in combination with known chemotherapeutics and their effect was studied on the growth and viability of a series of cell lines expressing low to high levels of TSPO. The CRAC ligand 7200432 (3-hydroxy-20-oxopregn-5-en-21-yl acetate) demonstrated significant inhibition of cellular growth and viability in a time- and dose-dependent manner. Moreover, this cytotoxic effect was dependent on TSPO expression, as its effects were magnified in high expressing TSPO cells and minimized in low-expressing TSPO cells. Modulatory profiling of 7200532 with the mitotic inhibitor paclitaxel or the topoisomerase inhibitor etoposide indicated that 7200532 can synergistically complement these chemotherapeutic agents to inhibit cell growth. In conclusion, we have performed lead generation screening of drugs targeted at the TSPO CRAC motif and identified a novel cytotoxic agent which can control cancer cell growth and may prove useful as a prototype compound for treatment of advanced metastases.[br][br](1) 1. Midzak A et al., J Biol Chem. 2011; 286:9875.[br][br]Sources of Research Support: This work was supported by a grant from the Canadian Institutes of Health Research (MOP102647) and a Canada Research Chair in Biochemical Pharmacology to V.P. A.M. was supported by postdoctoral fellowships from the Canadian Institutes of Health Research (TGF36110) and Le Fonds de la recherche du Quebec-sante. The Research Institute of MUHC was supported by a Center grant from Le Fonds de la recherche du Quebec-sante.[br][br]Nothing to Disclose: AM, RP, VP 2012-06-26T12:00:00 352DEF 2012-06-26T00:00:00 1899-12-30T12:00:00 1521 378 2738 OR35-4 OR44-01 Tuesday 2312 2012


2308 ENDO12L_OR35-5 ORAL SESSION: Neoplasia of Endocrine Tissues: Observations That Are Likely To Impact Clinical Management (11:15 AM-12:45 PM) Whole-Exome Sequencing Identifies the Genetic Landscape of Sporadic Parathyroid Adenomas Paul J Newey, Michael Andrew Nesbit, Andrew Rimmer, Rosie A Head, Paul T Christie, Caroline M Gorvin, Moustafa Attar, Lorna Gregory, Michael J Stechman, Radu Mihai, David Buck, Rajesh V Thakker University of Oxford, Oxford, UK; University of Oxford, Oxford, UK; University of Oxford, Oxford, UK; Oxford University Hospitals NHS Trust, Oxford, UK [bold]Background[/bold] The genetic etiology of sporadic parathyroid adenomas is poorly understood. To date, studies have focused on examining the contribution of candidate genes identified in familial tumor syndromes including [italic]MEN1[/italic], [italic]CDC73[/italic], [italic]RET[/italic], and [italic]CDKN1B[/italic]. Collectively, mutations in these genes occur in [lt]50% of parathyroid adenomas, indicating the involvement of additional tumor genes. [bold]Objective[/bold] To identify by whole-exome sequencing, genes commonly responsible for parathyroid tumorigenesis. [bold]Patients and Methods[/bold] 16 tumor-leucocyte pairs were obtained from post-menopausal women, without a family history of parathyroid disease, who had undergone curative surgery for primary hyperparathyroidism. Histology confirmed a single adenoma in all cases. Informed consent was obtained from individuals using approved protocols. Exome capture was performed using the SureSelect Human All Exon 50Mb Kit and sequencing undertaken using the Illumina HiSeq2000 platform. Somatic variants including single nucleotide variations (SNVs) and short insertions or deletions ([lt]50bp) were identified using an in-house variant calling algorithm. Somatic variants were validated by Sanger sequencing. [bold]Results[/bold] In total, 212 unique somatic variants were identified in the 16 tumors (median: 8/tumor; range: 2-110). All samples were adequately captured with [gt]90% of the targeted exonic regions represented by [gt]10 base reads. The majority of somatic variants were heterozygous non-synonymous SNVs resulting in missense amino acid changes. Somatic [italic]MEN1[/italic] mutations were identified in 6 out of 16 (37%) parathyroid adenomas. All tumors harboring somatic [italic]MEN1[/italic] mutations had additional variants (range: 2-10 additional variants/sample). Apart from [italic]MEN1[/italic], no gene was mutated in more than one tumor indicating the likely absence of additional major driver genes. However, mutations were identified in several genes that included [italic]MAPK8IP3[/italic] and [italic]SNRPB[/italic], and these may represent low frequency driver genes. [bold]Conclusions[/bold] Parathyroid tumors typically harbor a low number of somatic variants in keeping with their low proliferation rates and benign nature. [italic]MEN1[/italic] represents the major driver gene in sporadic parathyroid adenomas, although additional genetic events may be required for tumorigenesis. It is likely that multiple [apos]low frequency[apos] driver mutations occur in those parathyroid tumors that do not have [italic]MEN1[/italic] mutations. This study demonstrates the utility of whole-exome sequencing in defining the genetic landscape of parathyroid tumors.[br][br]Nothing to Disclose: PJN, MAN, AR, RAH, PTC, CMG, MA, LG, MJS, RM, DB, RVT 2012-06-26T12:15:00 352DEF 2012-06-26T00:00:00 1899-12-30T12:15:00 710 378 2739 OR35-5 OR44-01 Tuesday 2313 2012


2309 ENDO12L_OR35-6 ORAL SESSION: Neoplasia of Endocrine Tissues: Observations That Are Likely To Impact Clinical Management (11:15 AM-12:45 PM) Elevated Plasma POMC and AgRP Levels in Ectopic ACTH Tumors: Use in the Differential Diagnosis of ACTH- Dependent Cushing Syndrome Gabrielle Page-Wilson, Pamela U Freda, Thomas P Jacobs, Alexander G Khandji, Jeffery N Bruce, Kana Meece, Anne White, Sharon L Wardlaw Columbia University, College of Physicians [amp] Surgeons, New York, NY; Columbia University, College of Physicians [amp] Surgeons, New York, NY; Columbia University, College of Physicians [amp] Surgeons, New York, NY; University of Manchester, Manchester, UK The clinical presentations of Ectopic ACTH Syndrome (EAS) and Cushing[apos]s disease (CD) are often similar in patients with ACTH-dependent Cushing[apos]s Syndrome (CS) who have absent or small pituitary adenomas and no obvious ectopic tumor. Due to the frequency of pituitary incidentalomas and the limitations of imaging and biochemical testing, distinguishing between pituitary and ectopic causes of CS remains a challenge. While inferior petrosal sinus sampling (IPSS) is the gold-standard for discriminating between CD and EAS, it is a specialized procedure that is not widely available. Thus identifying novel diagnostic tools that capitalize on tumor specific differences in peptide synthesis and processing may prove useful. We previously demonstrated abnormal POMC processing and increased POMC precursor in patients with EAS vs CD with microadenomas. For the first time, we have observed marked plasma elevations of another neuropeptide, Agouti-related protein (AgRP) (3000 pg/ml) in a case of EAS from small cell lung cancer. We have now examined the utility of POMC measurements in a larger cohort of patients with ACTH-dependent CS and have evaluated plasma AgRP in parallel. POMC and AgRP were measured in peripheral blood samples from 32 patients who presented to our center for IPSS and had either no pituitary lesion or a microadenoma on MRI and in 14 healthy controls. POMC was measured using a specific monoclonal antibody based ELISA.[1] AgRP was measured by ELISA (R[amp]D). 22/32 patients had CD; 10/32 had EAS. Mean POMC was higher in EAS vs CD (67.3[plusmn]20.7 (SEM) vs 17.2[plusmn]1.5fmol/ml;p=0.001) vs controls (16.0[plusmn]1.5fmol/ml). All patients with POMC levels greater than 36fmol/ml had EAS (n=7), indicating that elevated POMC has a high positive predictive value for EAS. Mean AgRP levels were also higher in EAS vs CD (277[plusmn]77 vs 118 [plusmn] 6.1pg/ml;p=0.03). Of note, plasma AgRP was higher in CD vs controls (76.4[plusmn]5.4pg/ml;p=0.005). All subjects with an AgRP greater than 280pg/ml had EAS (n=3). When we characterized AgRP in the plasma of a patient with EAS by gel filtration and HPLC the majority of AgRP immunoactivity eluted in the position of full-length AgRP. Overall the presence of POMC [gt]36fmol/ml and/or AgRP [gt]280pg/ml identified 9/10 cases of EAS. These data demonstrate that measurement of POMC and AgRP in ACTH-dependent CS may facilitate the diagnosis of EAS. In addition, these data suggest that AgRP should be further evaluated as a potential neuroendocrine tumor marker.[br][br][1] SR Crosby, MF Stewart, JG Ratcliffe, A White. Direct Measurement of the Precursor of Adrenocorticotropin in Human Plasma by Two-Site Immunoradiometric Assay. J Clin Endocrinol Metab 67:1272, 1988.[br][br]Nothing to Disclose: GP-W, PUF, TPJ, AGK, JNB, KM, AW, SLW 2012-06-26T12:30:00 352DEF 2012-06-26T00:00:00 1899-12-30T12:30:00 924 378 2740 OR35-6 OR44-01 Tuesday 2314 2012


2310 ENDO12L_OR36-1 ORAL SESSION: Novel Molecular Mechanisms Affecting Gene Regulation by Steroid Receptors [amp] Breast [amp] Prostate Cancer Progression (11:15 AM-12:45 PM) Progestin Suppression of Mir-29 Potentiates Dedifferentiation of Breast Cancer Cells Via KLF4 Diana M Cittelly, Jessica Finlay-Schultz, Erin N Howe, Sunshine Daddario-Axlund, Peter Hendricks, Britta Jacobsen, Carol A Sartorius, Jennifer K Richer University of Colorado Denver Anschutz Medical Campus, Aurora, CO; University of Colorado Denver Anschutz Medical Campus, Aurora, CO The female hormone progesterone promotes the expansion of stem-like cancer cells in estrogen receptor (ER) and progesterone receptor (PR) positive breast tumors. The expanded tumor cells lose expression of ER and PR, express the tumor-initiating marker CD44, the progenitor marker cytokeratin 5 (CK5), and are more resistant to standard endocrine and chemotherapies. Virtually nothing is known about the mechanisms by which progestins reprogram more differentiated ER[sup]+[/sup]PR[sup]+[/sup]CK5[sup][ndash] [/sup]cells into progenitor ER[sup][ndash][/sup]PR[sup][ndash][/sup]CK5[sup]+[/sup] cells. Non-coding regulatory RNAs termed microRNAs (miRNAs) control various signaling networks important for breast cancer progression and metastasis. In this study, we investigated the role of miRNAs in progestin-mediated expansion of CK5[sup]+ [/sup]cell population. We recently identified miRNAs differentially expressed within 6 hours of progestin treatment of T47D cells. Among them, miR-29 family members (miR-29a, b and c) are significantly downregulated following progesterone (P4) stimulation, preceding the expansion of CK5[sup]+[/sup] CD44[sup]+[/sup] population in ER[sup]+[/sup]PR[sup]+[/sup] T47D and BT474 cell lines. Furthermore, the CD44[sup]hi/+[/sup] population expanded by P4 showed a significant downregulation of miR-29 family members compared to CD44[sup]low/-[/sup] T47D cells. We used lentiviral vectors (miR-ZIP) to stably inhibit miR-29 members and measured their ability to promote the expansion of CK5[sup]+[/sup] and CD44[sup]+[/sup] cells. We found that miR-29 members potentiate the expansion of CK5[sup]+[/sup] and CD44[sup]+[/sup] cells in response to P4, resulting in significantly increased mammosphere formation [italic]in vitro[/italic]. Using xenograft models, we show that miR-29a inhibition increases tumor growth. We demonstrate for the first time that miR-29 directly targets Kr[uuml]ppel-like factor 4 (KLF4), a transcription factor that is highly expressed in breast cancer stem cells and required for the maintenance of breast cancer stem cells. We show that P4 upregulates KLF4 by both direct transcriptional activation and indirect post-translational mechanisms (repressing miR-29) to exert maximal upregulation of this protein. Finally, we demonstrate that miR-29 repression and KLF4 upregulation are required for maximal expansion of CK5[sup]+[/sup] cells in response to progestins. These data provide a mechanism by which hormone control of potent reprogramming transcription factors through miRNAs regulate the [ldquo]stem-cell like population[rdquo] in breast tumors.[br][br]Sources of Research Support: DOD grants W81XWH-11-1-0101 awarded to DC and W81XWH-11-1-0210 awarded to CAS and JR.[br][br]Nothing to Disclose: DMC, JF-S, ENH, SD-A, PH, BJ, CAS, JKR 2012-06-26T11:15:00 362 2012-06-26T00:00:00 1899-12-30T11:15:00 212 379 2741 OR36-1 OR26-01 Tuesday 2315 2012


2311 ENDO12L_OR36-2 ORAL SESSION: Novel Molecular Mechanisms Affecting Gene Regulation by Steroid Receptors [amp] Breast [amp] Prostate Cancer Progression (11:15 AM-12:45 PM) DNA-Binding Cofactors Contribute to Cell Type[ndash]Specific PR Genomic Interaction in Normal and Malignant Breast Christine L Clarke, J Dinny Graham University of Sydney, Westmead Millennium Institute, Westmead, Australia The ovarian hormone progesterone is a critical regulator of diverse functions in the female reproductive system. In the normal breast, progesterone stimulates proliferation and development of lobular alveolar structures during pregnancy and in the luteal phase of the menstrual cycle. In breast cancer this influence takes on a more sinister aspect as exposure to progesterone and its synthetic analogues increases breast cancer risk. Progesterone effects are mediated via the nuclear progesterone receptor (PR), which binds to specific sequences in genomic DNA to regulate transcription of target genes. Where examined, the transcriptional responses to progesterone in different normal and malignant cell types are quite distinct, suggesting that differing patterns of PR genomic interaction may underlie this cell type specific response. We used genome wide PR ChIP sequencing to compare PR cistromes in two cell lines: T-47D breast cancer cells and MCF-10A immortalized normal breast cells stably expressing PR (AB32). We identified 6312 PR binding regions in T-47D and 8117 in AB32 cells. PR binding was correlated with transcriptional regulation in both cell lines, and 60% progestin regulated genes were associated with at least one PR binding region. The overlap between PR binding sites in the two cell lines was remarkably low, and this was reflected in a correspondingly low overlap in transcriptional response. Analysis of PR binding regions in both cell lines revealed a conserved progesterone response element that was similar in both cell lines. In addition, PR binding regions in T-47D were enriched for binding motifs for the pioneer factor FOXA1. No enrichment of FOXA1 binding motifs was detected in PR binding regions in AB32 cells. Further examination revealed that whereas robust expression of FOXA1 was detected in T-47D, the factor is extremely low to absent in AB32. Introduction of FOXA1 into AB32 cells resulted in a profound alteration in the transcriptional response to progesterone in these cells, demonstrating the potential for cell type specific factors to influence PR action. Enrichment of binding motifs for AP-1 and NF1 was observed only in AB32 cells, suggesting that these factors also contribute to cell type specific progesterone response. Our data suggest that cell-specificity of PR binding is determined by the coordinated effects of key binding cofactors.[br][br]Sources of Research Support: The National Health and Medical Research Council of Australia.[br][br]Nothing to Disclose: CLC, JDG 2012-06-26T11:30:00 362 2012-06-26T00:00:00 1899-12-30T11:30:00 1398 379 2742 OR36-2 OR26-01 Tuesday 2316 2012


2312 ENDO12L_OR36-3 ORAL SESSION: Novel Molecular Mechanisms Affecting Gene Regulation by Steroid Receptors [amp] Breast [amp] Prostate Cancer Progression (11:15 AM-12:45 PM) TWIST Represses Estrogen Receptor-[alpha] Expression Epigenetically by Recruiting the NuRD Protein Complex in Breast Cancer Cells Junjiang Fu, Lianmei Zhang, Tao He, Xiuli Xiao, Xiaoyan Liu, Li Wang, Luquan Yang, Manman Yang, Tiandan Zhang, Rui Chen, Jianming Xu Luzhou Medical College, Luzhou, China; Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX Loss of estrogen receptor [alpha] (ER[alpha]) expression and gain of TWIST expression in breast tumors correlate with increased disease recurrence and metastasis and poor disease-free survival. However, the molecular and functional regulatory relationship between TWIST and ER[alpha] are unclear. In this study, we found TWIST was associated with a chromatin region in intron 7 of the human [italic]ESR1[/italic] gene coding for ER[alpha]. This association of TWIST efficiently recruited the nucleosome remodeling and deacetylase (NuRD) repressor complex to this region, which subsequently decreased histone H3K9 acetylation, increased histone H3K9 methylation and repressed [italic]ESR1[/italic] expression in breast cancer cells. In agreement with these molecular events, TWIST expression was inversely correlated with ER[alpha] expression in both breast cancer cell lines and human breast ductal carcinomas. Forced expression of TWIST in TWIST-negative and ER[alpha]-positive breast cancer cells such as T47D and MCF-7 cells reduced ER[alpha] expression, while knockdown of TWIST in TWIST-positive and ER[alpha]-negative breast cancer cells such as MDA-MB-435 and 4T1 cells increased ER[alpha] expression. Furthermore, inhibition of histone deacetylase (HDAC) activity including the one in NuRD complex significantly increased ER[alpha] expression in MDA-MB-435 and 4T1 cells. HDAC inhibition together with TWIST knockdown will not further increased ER[alpha] expression in 4T1 and MDA-MB-435 cells. These results demonstrate that TWIST/NuRD represses ER[alpha] expression in breast cancer cells. Therefore, TWIST may serve as a potential molecular target for converting ER[alpha]-negative breast cancers to ER[alpha]-positive breast cancers, allowing these cancers to restore their sensitivity to endocrine therapy with selective ER[alpha] antagonists such as tamoxifen and raloxifene.[br][br](1)Yang J, et al. Cell 2004; 117: 927-39. (2)Fu J, et al. Cell Res 2011; 21: 275-89. (3)Qin Q, et al. Cell Res 2012; 22: 90-106.[br][br]Sources of Research Support: This work was supported by the National Natural Science Foundation of China (81172049), the Scientific Research Foundation of Education Bureau of Sichuan Province (10ZA038), a Key Research Project of Luzhou City (2011-S-30) and a Starting Package of Luzhou Medical College to J.F., and the National Institutes of Health grant R01 CA112403 to J.X.[br][br]Nothing to Disclose: JF, LZ, TH, XX, XL, LW, LY, MY, TZ, RC, JX 2012-06-26T11:45:00 362 2012-06-26T00:00:00 1899-12-30T11:45:00 241 379 2743 OR36-3 OR26-01 Tuesday 2317 2012


2313 ENDO12L_OR36-4 ORAL SESSION: Novel Molecular Mechanisms Affecting Gene Regulation by Steroid Receptors [amp] Breast [amp] Prostate Cancer Progression (11:15 AM-12:45 PM) GATA2 as a Novel Therapeutic Target for Castration-Resistant Prostate Cancer (CRPC) Bin He, Sue Anne Chew, Vijay Kumar Eedunuri, Chuandong Geng, Diego Jacinto Bedoya, Ashesh Shah, Nicholas Mitsiades Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Adrienne Helis Malvin Medical Research Foundation, New Orleans, LA [bold]Background: [/bold]In castration-resistant prostate cancer (CRPC), androgen receptor (AR) signaling persists despite low circulating testosterone levels, due to various mechanisms that include overexpression and/or mutation of AR and/or its coactivators, as well as other signaling aberrations that promote ligand-independent AR activation (LIAA). Although recent advances in AR targeting with abiraterone (androgen synthesis inhibitor) and MDV3100 (second generation AR antagonist) have improved clinical outcomes, resistance eventually emerges again with reactivated AR due to LIAA.[br][bold]Rationale:[/bold] There is an unmet need to effectively target AR expression and LIAA in CRPC. In this study, we investigated the role of the transcription factor GATA2 in AR expression and function in prostate cancer (PC) and tested the potential of GATA2 inhibitors as novel therapeutics for CRPC.[br][bold]Methods:[/bold] Cell viability of PC cell lines after treatment with small interfering RNA (siRNA) or small molecule inhibitors (SMIs) [italic]in vitro[/italic] was assessed by MTT. AR expression was assessed by immunoblotting and RTqPCR. AR function was assessed by RTqPCR for several known AR target genes. K7174, a previously described GATA2 SMI, was provided by Kowa (Tokyo, Japan). In order to identify additional GATA2 SMIs, we interrogated the Connectivity Map (Broad Institute, Cambridge, MA), a database of genome-wide transcriptional expression data from cultured human cells treated with bioactive small molecules, using a gene expression signature of LNCaP cells treated with GATA2 siRNA.[br][bold]Results: [/bold]GATA2 is required for AR expression and function in a variety of PC cell lines. siRNA-mediated GATA2 knockdown significantly suppressed the growth of both androgen-dependent (LNCaP, VCaP, and LAPC4) and castration-resistant (22Rv1, C4-2, and LNCaP-abl) PC cells, while it had little effect on the GATA2(-)/AR(-) PNT1A and PC3 cells. Our [italic]in silico[/italic] screening identified Pyrvinium Pamoate (PP), a compound previously reported to non-competitively inhibit AR via an unknown mechanism, as a potential GATA2 SMI. Both PP and K7174 inhibited AR expression and function, as well as growth of a variety of GATA2(+)/AR(+) PC cell lines. PP or K7174 in combination with MDV3100 (Medivation, San Francisco, CA) synergistically suppressed AR function and cell growth in LNCaP cells.[br][bold]Conclusions: [/bold]GATA2 is essential for AR expression and function, and for growth of PC cells. GATA2 SMIs hold promise as novel CRPC therapeutics.[br][br]Sources of Research Support: The authors acknowledge the joint participation by Adrienne Helis Malvin Medical Research Foundation through its direct engagement in the continuous active conduct of medical research in conjunction with Baylor College of Medicine. B.H. is supported by NIH/NIDDK grant 5K01DK081446. N.M. is a Dan L. Duncan Scholar and a Caroline Wiess Law Scholar at Baylor College of Medicine and his work is also supported by the Conquer Cancer Foundation of the American Society of Clinical Oncology (ASCO) Career Development Award and by the Prostate Cancer Foundation.[br][br]Nothing to Disclose: BH, SAC, VKE, CG, DJB, AS, NM 2012-06-26T12:00:00 362 2012-06-26T00:00:00 1899-12-30T12:00:00 1679 379 2744 OR36-4 OR26-01 Tuesday 2318 2012


2314 ENDO12L_OR36-5 ORAL SESSION: Novel Molecular Mechanisms Affecting Gene Regulation by Steroid Receptors [amp] Breast [amp] Prostate Cancer Progression (11:15 AM-12:45 PM) SUMOylation Governs the Anti-Proliferative Properties and Stress Responsiveness of Glucocorticoid Receptor in Human Embryonal Kidney Cells Ville Paakinaho, Sanna Kaikkonen, Harri Makkonen, Jorma J Palvimo University of Eastern Finland, Kuopio, Finland Endocrine system responds to stress by secreting glucocorticoids. Glucocorticoids are also among the key drugs for treating diseases ranging from hematological cancers to inflammatory states. In addition to activating the glucocorticoid receptor (GR), the glucocorticoids influence posttranslational modifications of the receptor, including its SUMOylation status. To study how GR SUMOylation influences the activity of endogenous glucocorticoid target genes in a normal chromatin environment, we compared glucocorticoid-induced gene expression in isogenic human embryonal kidney (HEK293) cells stably expressing wild-type GR (GRwt) or SUMOylation-deficient GR (GR3KR). Genome-wide gene expression profiling of these cells indicates that dexamethasone (dex) regulated more genes in GR3KR-expressing than in GRwt-expressing cells (673 vs. 444 genes, p[lt]0.001). According to Ingenuity Pathway Analysis, the dex-regulated genes whose expression differs between the cell lines are significantly associated with cell cycle control, cell proliferation and differentiation pathways. These genes include I[kappa]B[alpha] and CDK inhibitors [italic]CDKN1A[/italic] (p21) and [italic]CDKN1C[/italic] (p57). Furthermore, genes of the Cip/Kip and Ink4 family of CDK inhibitors, except for p21, are expressed at a markedly higher level in GRwt cells than in GR3KR cells. The GR3KR-expressing cells also proliferated more rapidly than the GRwt-expressing cells and the antiproliferative effect of dex was less pronounced on the SUMOylation-deficient GR-expressing cells. Since the activity of SUMOylation pathway is regulated by cell stress, we also studied the effect of anti-inflammatory prostaglandin 15d-PGJ[sub]2[/sub], a mild oxidative stress inducer, on these cell lines. Interestingly, the glucocorticoid-dependent expression of genes, such as p57, S100P and I[kappa]B[alpha], was attenuated in response to 15d-PGJ[sub]2[/sub] exposure in GRwt cells, whereas this was not the case in GR3KR cells. Moreover, the inhibition of I[kappa]B[alpha] expression by 15d-PGJ[sub]2[/sub] was much weaker in the GR3KR cells than in the GRwt cells. ChIP assays showed that the loading of GR onto p57, S100P and I[kappa]B[alpha] regulatory regions was decreased in GRwt cells but not in GR3KR cells. In sum, our results suggest that SUMOylation regulates the activity of GR in a target gene and gene program selective fashion, playing a key role in the regulation of GR target genes and programs influencing cell growth. Moreover, SUMOylation seems regulate the responsiveness of GR to cell stress.[br][br]Nothing to Disclose: VP, SK, HM, JJP 2012-06-26T12:15:00 362 2012-06-26T00:00:00 1899-12-30T12:15:00 479 379 2745 OR36-5 OR26-01 Tuesday 2319 2012


2315 ENDO12L_OR36-6 ORAL SESSION: Novel Molecular Mechanisms Affecting Gene Regulation by Steroid Receptors [amp] Breast [amp] Prostate Cancer Progression (11:15 AM-12:45 PM) Role of Lysine Deacetylases in Glucocorticoid Receptor-Mediated Transcriptional Activation Vineela Kadiyala, Wana Mathieu, Lingling An, Catharine L Smith University of Arizona, Tucson, AZ; University of Arizona, Tucson, AZ Lysine deacetylases (KDACs) are known targets of a group of drugs being evaluated in many clinical and pre-clinical trials for use in the treatment of cancer and other diseases. Hence it is essential to understand the role of KDACs in signaling and transcription to reveal the full cellular effects of KDAC inhibitors (KDACi). The goal of our study is to elucidate how KDACs and protein acetylation regulate the glucocorticoid (GC) signaling pathway. Glucocorticoid receptor (GR) is known to associate with class I KDACs to regulate the expression of a few target genes. However, the effect of KDAC inhibition on the GR-regulated transcriptome is unknown. To illustrate this we carried out expression profiling studies in a GC responsive hepatoma-derived cell line. The results show that the class I-selective KDACi, VPA, has a profound impact on the GR-regulated hepatic transcriptome. VPA treatment alone mimics GC signaling at some GR target genes and cooperates with GC to activate a small number of genes. However, the predominant effect of VPA, seen in more than 50% of the GR target genes, is impairment of normal GR-mediated activation. This suggests that KDACs play a significant role in facilitating GC signaling. Co-immunoprecipitation assays and western blotting have shown that VPA does not disrupt the GR-HSP90 complex, indicating that its inhibitory effect on GR transactivation is not through impaired GR processing. We have also determined that apicidin, a structurally distinct class I-selective KDACi, has similar effects on GR-regulated transcription as that of VPA. In contrast, valpromide, a structural analog of VPA without KDACi activity, has different effects. This strongly suggests that VPA impairs GR transactivation through inhibition of class I KDACs. KDAC depletion through siRNAs was used to identify individual KDACs involved in GR transactivation. Our results show that depletion of KDAC1 fully or partially mimics the effects of VPA at some but not all of the impaired GR target genes. This finding has led us to conclude that, in addition to KDAC1, other class I KDACs facilitate transcription at GR target genes. Current studies are geared towards identifying these additional class I KDACs and elucidating the effect of KDAC activity on GR recruitment to GR target genes.[br][br]Sources of Research Support: Bio5 Institute; Rescorp; NSF MCB-1122088.[br][br]Nothing to Disclose: VK, WM, LA, CLS 2012-06-26T12:30:00 362 2012-06-26T00:00:00 1899-12-30T12:30:00 687 379 2746 OR36-6 OR26-01 Tuesday 2320 2012


2316 ENDO12L_OR37-1 ORAL SESSION: Thyroid Cancer: Observations Likely To Impact Clinical Management (11:15 AM-12:45 PM) Inhibition of Src Kinase with Dasatinib Blocks Thyroid Cancer Growth and Metastasis Christine M Chan, Xia Jing, Laura A Pike, Qiong Zhou, Dong-Jun Lim, Gregory S Lund, Vibha Sharma, Bryan R Haugen, Rebecca E Schweppe University of Colorado School of Medicine, Aurora, CO; University of Colorado School of Medicine, Aurora, CO There are currently no effective therapies for patients with advanced thyroid cancer, which includes patients diagnosed with advanced papillary thyroid cancer (PTC) and anaplastic thyroid cancer (ATC). Much attention has been devoted to the mitogen activated protein kinase (MAPK) pathway as a therapeutic target due to the large percentage of activating mutations in this pathway, yet additional therapeutic targets are needed for the effective long-term treatment of advanced thyroid cancer patients. Src Family Kinases (SFKs) are overexpressed and activated in numerous tumor types and have emerged as a promising therapeutic target, especially in relation to metastasis. We recently discovered that the Src target, FAK, is overexpressed and phosphorylated in patient thyroid tumor samples, as well as cell lines derived from advanced thyroid cancer patients. We further demonstrated that the growth of a subset of thyroid cancer cells is susceptible to treatment with the Src inhibitors, saracatinib and dasatinib. Here, we show that inhibition of Src with 10 or 50 nM dasatinib results in cell cycle arrest (8-22%) and increased apoptosis (2-4 fold) in a subset of thyroid cancer cells (BCPAP, SW1736, K1), and that treatment with 10 or 50 nM dasatinib results in decreased colony formation in the BCPAP and K1 cells (36-85% inhibition), but not the 8505C cells. Immunoblotting showed that c-Src and Lyn are the only SFK members expressed in thyroid cancer cells (C643, TPC1, SW1736, BCPAP, K1, 8505C), and that c-Src, but not Lyn, is activated. Expression of a dasatinib-resistant Src gatekeeper mutant confirmed the selectivity of dasatinib, and that c-Src is likely the primary mediator of cellular growth in response to Src inhibitor treatment. Furthermore, treatment with dasatinib blocked PTC tumor growth in an orthotopic model by [gt] 90% (p=0.0014). In a novel experimental metastasis model (bone, brain), adjuvant treatment with dasatinib strongly inhibited metastasis (p=0.016). Importantly, treatment with dasatinib in the metastatic model was stopped on day 56, and dasatinib-treated mice exhibited a significantly increased overall survival ([sim]2-fold) when compared to vehicle treated mice (p = 0.0015). Taken together, these data provide strong evidence that Src is a key mediator of thyroid cancer growth, and indicate that inhibition of Src with dasatinib represents a promising therapeutic strategy to block both primary tumor growth and metastasis in thyroid cancer patients.[br][br]Sources of Research Support: National Cancer Institute grant K12-CA086913 (RE Schweppe), and NIH 5 T32 DK063687 (CM Chan).[br][br]Nothing to Disclose: CMC, XJ, LAP, QZ, D-JL, GSL, VS, BRH, RES 2012-06-26T11:15:00 Theater B 2012-06-26T00:00:00 1899-12-30T11:15:00 2202 380 2747 OR37-1 OR41-01 Tuesday 2321 2012


2317 ENDO12L_OR37-2 ORAL SESSION: Thyroid Cancer: Observations Likely To Impact Clinical Management (11:15 AM-12:45 PM) SKI-606, a Src Inhibitor, Reduces Tumor Growth, Invasion, and Distant Metastasis in a Mouse Model of Thyroid Cancer Won Gu Kim, Celine J Guigon, Laura Fozzatti, Jeong Won Park, Changxue Lu, Mark C Willingham, Sheue-Yann Cheng National Cancer Institute, Bethesda, MD; Wake Forest University, Winston-Salem, NC Currently, no effective treatment for patients with radioiodine-refractory, advanced differentiated thyroid cancer is available. Novel treatment modalities are being actively sought. Aberrant activation of the Src pathway is associated with aggressive thyroid cancer. We hypothesized that inhibition of the Src pathway could prevent thyroid cancer progression and distant metastasis. The [italic]Thrb[sup]PV/PV[/sup][/italic] mouse, harboring a knock-in dominantly negative mutation of the thyroid hormone receptor [beta] gene (denoted as [italic]ThrbPV[/italic]), spontaneously develops metastatic thyroid cancer. In this mouse model, we found that an over-activated Src pathway promotes thyroid carcinogenesis. For this study, we evaluated the effects of SKI-606, a potent inhibitor of Src, on thyroid carcinogenesis of [italic]Thrb[sup]PV/PV[/sup]Pten[sup]+/-[/sup][/italic] mice, which were generated by haploid deficiency of the tumor suppressor gene [italic]Pten[/italic]. These mice exhibited a more aggressive cancer phenotype with decreased survival, thus shortening the time for preclinical studies. SKI-606 treatment effectively inhibited aberrant activation of Src and its downstream targets to markedly inhibit the growth of thyroid tumor by 37%, thereby significantly increasing the survival of SKI-606-treated mice by 1.2 months as compared with vehicle-treated mice. Src inhibition reduced cell proliferation (38%) as evidenced by the 50% reduction in Ki-67 nuclear staining. SKI-606 treatment decreased the expression of key regulators of cell cycle progression, including cyclin B1 (85%), cyclin D1 (57%), cyclin E (57%), CDK4 (93%), CDK6 (38%), phosphorylated-Rb (75%), and E2F1 (93%). Importantly, SKI-606 treatment decreased vascular invasion (26%), and lung metastasis (45%) of thyroid cancer by inhibiting epithelial-mesenchymal transition via increased expression of E-cadeherin (2.2-fold) and decreased expression of vimentin (94%), slug (81%), MMP-2 (53%), and MMP-9 (89%). SKI-606 treatment also dramatically prevented de-differentiation by maintaining the normal follicular structure and expression of PAX8 and NIS differentiation markers via down-regulation of MAPK pathways. This preclinical study showed that for the first time, blocking of Src activity delays tumor progression and inhibits metastasis. Thus, Src and its downstream effectors are potential molecular targets for treatment of thyroid cancer. Importantly, oral SKI-606 is a promising treatment strategy for single-drug therapy or in combination with radioiodine therapy to treat refractory thyroid cancer.[br][br]Sources of Research Support: The present research was supported by the Intramural Research Program at the Center for Cancer Research, National Cancer Institute, National Institutes of Health.[br][br]Nothing to Disclose: WGK, CJG, LF, JWP, CL, MCW, S-YC 2012-06-26T11:30:00 Theater B 2012-06-26T00:00:00 1899-12-30T11:30:00 98 380 2748 OR37-2 OR41-01 Tuesday 2322 2012


2318 ENDO12L_OR37-3 ORAL SESSION: Thyroid Cancer: Observations Likely To Impact Clinical Management (11:15 AM-12:45 PM) Next Generation Sequencing Reveals the Specific Expression of microRNA Isoforms in Normal Thyroid and Papillary Thyroid Carcinoma Michal Swierniak, Anna Wojcicka, Malgorzata Czetwertynska, Elzbieta Stachlewska, Wieslaw Wiechno, Barbara Gornicka, Stefano Volinia, Marek Krawczyk, Albert de la Chapelle, Krystian Jazdzewski Medical University of Warsaw, Warsaw, Poland; Ohio State University, Columbus, OH; Medical University of Warsaw, Warsaw, Poland; Medical University of Warsaw, Warsaw, Poland; Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland; Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland; Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice, Poland; Universita degli Studi, Ferrara, Italy To elucidate the role of miRs in papillary thyroid carcinoma (PTC) we employed the next-generation sequencing (SOLID) to analyze the miRNA profiles of PTC tumors (n=19), unaffected tissue adjacent to tumors (n=19), and normal (control) thyroid tissue (n=14). We received sequences of miRNAs present in the thyroid tissue together with a corresponding number of reads reflecting their expression levels, which were normalized to total reads available for all miRs (RPM, reads per million).[br]400 miRs deposited in miRbase were found to be significantly expressed in the thyroid tissue (RPM[gt]5 in [gt]50% of samples). Among them, 119 were significatly deregulated in PTC tumors compared to normal thyroid (FDR[lt]0.05), including 25 strongly upregulated (fold change [gt]2), and 58 strongly downregulated (fold change [lt]0.5). The list of top deregulated miRs comprises miR-146b, -221, -7, -222, -21, -34a (fold change 21, 8.3, 0.14, 5.5, 4.9, 3.2, respectively), confirming the previous microarray profiling. In addition, top-deregulated miRs include several miRs derived from the passenger strand of miRNA precursors, e.g. miR-146b*, -7*, -221*, -34a*, -222*, -545* (fold change 17.6, 0.13, 5.1, 4.0, 3.6, 3.2).[br]Most mature miRs exist in variable length variants, called isomiRs. Interestingly, our results indicate that 85% of the most abundant miRNA isoforms differ from the standard reference sequence deposited in the miRBase. Moreover, the standard miR sequence was completely absent in 34% of miRs expressed in PTC tumors.[br]Among the identified isomiRs, numerous molecules were found to differ at their 5[apos]end which is of particular interest as such miRs have different seed sequences, i.e potentially target different transcripts. For the highly deregulated miR-146b we detected a significant expression of six isoforms, of which two varied at the 5[apos]end (1nt difference). All the isomiRs were deregulated in PTC tumors compared to normal thyroid tissue (14.7 to 29 fold change, FDR[lt]0.002). Based on the TargetRank prediction, the two 5[apos]end isomiRs of miR-146b share only 15% of target genes.[br]Our results provide a comprehensive view of the miRNA transcriptome of the thyroid gland and reveal numerous miRNA variants differentially expressed in PTC, with a fold change up to 29. We show that almost all miRNAs exhibit isoforms of variable length and thus potentially distinct function. Elucidation of miR expression profiles will lead to identification of new targets for diagnosis and treatment of PTC.[br][br]1) Jazdzewski K, Liyanarachchi S, Swierniak M, Pachucki J, Ringel MD, Jarzab B, de la Chapelle A 2009 Polymorphic mature microRNAs from passenger strand of pre-miR-146a contribute to thyroid cancer. Proc Natl Acad Sci USA 106:1502-1505. 2) de la Chapelle A, Jazdzewski K 2011 MicroRNAs in thyroid cancer. J Clin Endocrinol Metab 96:3326-3336.[br][br]Sources of Research Support: This work was supported by Polish Ministry of Science and Higher Education Grants NN401 584838 and 0150/P01/2010/70, National Cancer Institute Grants P30 CA16058 and P01 CA124570, and Foundation for Polish Science (program FOCUS).[br][br]Nothing to Disclose: MS, AW, MC, ES, WW, BG, SV, MK, AdlC, KJ 2012-06-26T11:45:00 Theater B 2012-06-26T00:00:00 1899-12-30T11:45:00 1857 380 2749 OR37-3 OR41-01 Tuesday 2323 2012


2319 ENDO12L_OR37-4 ORAL SESSION: Thyroid Cancer: Observations Likely To Impact Clinical Management (11:15 AM-12:45 PM) The Interference of Anti-Tg Antibodies with Diagnostic Value of Thyroglobulin in Washout Fluid from Fine-Needle Aspiration Biopsy of Neck Nodes for Detecting Metastasis of Papillary Thyroid Carcinoma MinJi Jeon, JiMin Han, JeongHyun Lee, TaeYong Kim, WonBae Kim, YoungKee Shong Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea; Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea [bold]Background[/bold][br]Thyroglobulin (Tg) measurement from fine needle aspiration biopsy washes is recommended in patients with ultrasonographically suspicious metastatic lymph nodes of papillary thyroid carcinoma (PTC). However, the cutoff value of Tg from fine needle aspiration biopsy washes (FNA-Tg) and interference of serum anti-Tg antibodies (TgAb) with FNA-Tg have not been clearly identified.[br][bold]Methods[/bold][br]We reviewed PTC patients with FNA-Tg result who initially underwent total thyroidectomy with remnant ablation during 2-year period. FNA-Tg was performed on ultrasonographically suspicious metastatic neck nodes equal or greater than 5mm. Serum Tg, TgAb and TSH measurements were performed with FNAB procedure and the latest stimulated Tg value was retrieved. TgAb value greater than 60 IU/ml was considered positive.[br][bold]Results[/bold][br]Two hundred sixteen lymph nodes from 173 patients were evaluated. The final histopathology was available from 84 nodes and 81 (96%) of them showed malignant nodes. Seven nodes from nodes with benign cytology were turned out to be metastatic lymph nodes in pathology and they were all showed high FNA-Tg (FNA-Tg 768-19,100 ng/ml). FNA-Tg level were lower in nodes with positive TgAb than in nodes with negative TgAb (P [lt]0.05). The lowest FNA-Tg levels from malignant nodes with negative TgAb were 14.5 ng/mL. However, among the 14 nodes with positive TgAb, 5 metastatic nodes showed below 14.5 ng/mL of FNA-Tg. Besides, 2 of them revealed below 1ng/mL of both FNA-Tg and stimulated Tg.[br][bold]Conclusion[/bold][br]High TgAb concentration interfered with FNA-Tg. TgAb causes falsely low FNA-Tg, below the analytical sensitivity of assay and mask the presence of malignancy. However low the FNA-Tg is, we could not exclude malignancy if the TgAb level is high.[br][br]Nothing to Disclose: MJ, JH, JL, TK, WK, YS 2012-06-26T12:00:00 Theater B 2012-06-26T00:00:00 1899-12-30T12:00:00 1207 380 2750 OR37-4 OR41-01 Tuesday 2324 2012


2320 ENDO12L_OR37-5 ORAL SESSION: Thyroid Cancer: Observations Likely To Impact Clinical Management (11:15 AM-12:45 PM) Suspicious Cervical Lymph Nodes Detected after Thyroidectomy Infrequently Show Clinically Significant Growth in Papillary Thyroid Cancer Patients Eyal Robenshtok, Stephanie Fish, Ariadne Bach, R Michael Tuttle Memorial Sloan-Kettering Cancer Center, New York, NY; Memorial Sloan-Kettering Cancer Center, New York, NY [bold]Background:[/bold] The risk of locoregional recurrence in papillary thyroid cancer (PTC) patients ranges from 15% to 25%, and occurs mostly in the cervical lymph nodes. The impact and the natural history of small volume recurrent disease detected by current more sensitive surveillance is unclear. This study was designed to describe the structural changes over time in abnormal lymph nodules identified on routine surveillance neck ultrasounds (US) after thyroidectomy.[br][bold]Patients and Methods: [/bold]We retrospectively analyzed the outcome of 166 patients with suspicious cervical nodes outside the thyroid bed with at least one abnormal feature other than size (including hypervascularity, abnormal shape, hypoechogenicity, absence of hilum, calcifications, cystic changes, and other changes as previously described), who were followed for more than one year using serial US. Clinicopathologic and sonographic characteristics were analyzed as possible predictors for nodes growth. The primary endpoint was lymph node growth during follow-up.[br][bold]Results[/bold]: Most patients had classical PTC (85%) and an intermediate risk of recurrence according to the ATA classification (77%). The median size of the nodes was 1.3cm (range 0.5-2.7) in largest diameter, with all nodes having at least one abnormal characteristic, and 2 or more features in 70%. Almost all included LN were in the lateral neck, most in levels 4 and 3 (58% and 43% respectively) and some in levels 2(16%), 5 (4%) and 6 (1.3%).[br]After a follow-up of a median of 3.5 years, only 20% (33/166) grew 3mm or more and 9% (15/166) grew 5mm or more. None of the clinical and sonographic features were predictive of nodes growth (positive predictive value range 0.21-0.57). Seven patients had FNA proven metastatic LN at baseline, with a median size of 1.7cm and median follow-up of 8 years. All of these nodes were stable in size during the study period. There were no local complications (nerve damage or invasion to adjacent tissue) related to the abnormal nodes, and there was no disease related mortality during the study period.[br][bold]Conclusions[/bold]: Suspicious cervical lymph nodes in patients with PTC often remain stable for long periods of time and in properly selected patients can be safely followed using serial ultrasounds.[br][br]Disclosures: RMT: Ad Hoc Consultant, Genzyme Corporation; Clinical Researcher, Genzyme Corporation. Nothing to Disclose: ER, SF, AB 2012-06-26T12:15:00 Theater B 2012-06-26T00:00:00 1899-12-30T12:15:00 914 380 2751 OR37-5 OR41-01 Tuesday 2325 2012


2321 ENDO12L_OR37-6 ORAL SESSION: Thyroid Cancer: Observations Likely To Impact Clinical Management (11:15 AM-12:45 PM) A Randomized Controlled Trial of a Computerized Decision Aid on Adjuvant Radioactive Iodine Treatment for Patients with Early-Stage Papillary Thyroid Cancer: Impact on Medical Knowledge and Decisional Conflict Anna M Sawka, Sharon Straus, Lorne Rotstein, James D Brierley, Richard W Tsang, Sylvia Asa, Phillip Segal, Catherine Kelly, Afshan Zahedi, Jeremy Freeman, Philip Solomon, Jennifer Anderson, Kevin E Thorpe, Amiram Gafni, Gary Rodin, David P Goldstein University of Toronto, Toronto, Canada; University of Toronto, Toronto, Canada; University of Toronto, Toronto, Canada; University of Toronto, Toronto, Canada; University of Toronto, Toronto, Canada; Keenan Research Centre, Li Ka Shing Knowledge Institute of St Michael[apos]s Hospital and the Dalla Lana School of Public Health, Toronto, Canada; McMaster University, Hamilton, Canada; University of Toronto, Toronto, Canada [bold]Purpose: [/bold]Decision-making on adjuvant radioactive iodine (RAI) treatment for early stage papillary thyroid cancer (PTC) is complex, due to uncertainties in medical evidence. Using a parallel, 2-arm, randomized controlled trial design, we examined the impact of a patient-directed computerized decision aid (DA) on the medical knowledge and decisional conflict in patients with early stage PTC considering the choice of RAI treatment or no RAI treatment. This DA describes the rationale, possible risks and benefits, as well as the uncertainty of related medical evidence.[br][bold]Patients and Methods: [/bold]We recruited 74 patients with early stage PTC, after thyroidectomy. Participants were assigned, using 1:1 central computerized randomization, to the DA group with usual care (intervention) or usual care alone (control). Medical knowledge about PTC and RAI treatment (the primary outcome), as well as decisional conflict (a secondary outcome), were measured using validated questionnaires, and the respective scores were compared between groups.[br][bold]Results: [/bold]Consistent with the gender distribution of PTC, 83.8% (62/74) of the subjects were women, and the mean age of participants was 45.8 years (range 19 to 79 years). Medical knowledge about PTC and RAI treatment was significantly greater in the DA group than in the control group (p[lt]0.001). Decisional conflict on treatment choice was also significantly reduced in the DA group (p[lt]0.001).[br][bold]Conclusions: [/bold]A computerized DA significantly improves medical knowledge and reduces decisional conflict in patients with early stage PTC considering adjuvant RAI treatment. Decision aids may be particularly useful for patients facing decisions subject to uncertainty of medical evidence.[br][br]Sources of Research Support: This study was supported by an operating grant from the Ontario Ministry of Health and Long-term Care (Alternate Funding Plan - Innovation Fund). Anna Sawka has been supported, in part, by a New Investigator Grant from the Canadian Institutes of Health Research (CNI-80701). Anna Sawka currently holds a Chair in Health Services Research from Cancer Care Ontario, funded by the Ontario Ministry of Health and Long-term Care. Sharon Straus holds a Tier 1 Canada Research Chair.[br][br]Nothing to Disclose: AMS, SS, LR, JDB, RWT, SA, PS, CK, AZ, JF, PS, JA, KET, AG, GR , DPG 2012-06-26T12:30:00 Theater B 2012-06-26T00:00:00 1899-12-30T12:30:00 76 380 2752 OR37-6 OR41-01 Tuesday 2326 2012


2322 ENDO12L_OR38-1 ORAL SESSION: Vitamin D: Supplementation, Metabolism [amp] Function (11:15 AM-12:45 PM) Incidence of Hypercalciuria and Hypercalcemia during a Vitamin D Trial in Postmenopausal Women Vinod Yalamanchili, John Gallagher Creighton University Medical Center, Omaha, NE Background: Long-term use of vitamin D and calcium supplements is common in older women. But there is no long-term data on hypercalcemia and hypercalciuria on supplements. In the WHI study there was a significant increase in renal stones in women on 400 IU/d dose of vitamin D3 and 1000mg extra calcium after 7 years but no biochemical data. We collected serum and urine calcium data every 3 months during a one-year trial of different doses of vitamin D.[br]Methods: Participants in this study were 163 Caucasian postmenopausal women, ages 57-85 years, randomized to one of the several doses of vitamin D3 - 400, 800, 1600, 2400, 3200, 4000, 4800 IU/day or placebo. Calcium intake was increased to 1200-1400mg/d using calcium citrate from a baseline average of 691 mg/d. The main inclusion criterion was vitamin insufficiency -serum 25OHD [lt] 20ng/ml (Diasorin assay). Exclusion criteria were illness or medications known to affect vitamin D metabolism. Serum and urine calcium were measured at baseline and every 3 months for one year. Multiple regression including age, dose, calcium intake, weight, serum 25OHD was used to analyze serum and urine calcium results separately. Definition of hypercalcemia was a value [gt] normal range (8.9-10.3 mg/dl) and hypercalciuria a 24h urine calcium [gt] normal (300mg). Any abnormal event was verified after one week and if high values continued then calcium supplements were reduced or vitamin D was stopped.[br]Results: Mean baseline serum 25OHD was 15.6 ng/ml and increased on the highest dose of vitamin D to 45 ng/ml. Mean baseline serum calcium was 9.47 mg/dl and increased to 9.52 mg/dl; mean 24-hr urine calcium was 142 mg and increased to 186 mg. Final 24h urine calcium increased with vitamin D dose. There were 88 episodes of hypercalciuria and 25 episodes of hypercalcemia. There was no significant association between episodes of hypercalcemia or hypercalciuria and vitamin D dose or serum 25OHD and no kidney stones. Prolonged hypercalciuria led to discontinuation of calcium in 2 and vitamin D in 1 subject.[br]Conclusion: About 33 percent of subjects had an episode of hypercalciuria and [sim]10 percent an episode of hypercalcemia on vitamin D and calcium. These changes were not related to vitamin D dose. If these changes occur frequently with long-term use it could explain the increase in kidney stones seen in the WHI study. Limitation of calcium supplements and measurement of 24h urine calcium is advisable with long-term use of vitamin D and calcium.[br][br]Sources of Research Support: National Institute of Aging.[br][br]Nothing to Disclose: VY, JG 2012-06-26T11:15:00 332 2012-06-26T00:00:00 1899-12-30T11:15:00 2065 381 2753 OR38-1 OR07-01 Tuesday 2327 2012


2323 ENDO12L_OR38-2 ORAL SESSION: Vitamin D: Supplementation, Metabolism [amp] Function (11:15 AM-12:45 PM) Evaluating Vitamin D Repletion Regimens and Their Relationship to Changes in Vitamin D Serum Levels in Veteran Patients Sheryl Aterrado, Gregory Ono, Shawn Kanehira-Mar, Joy Meier, Arthur Swislocki Department of Veterans Affairs Northern California Healthcare System, Mather, CA; Department of Veterans Affairs Northern California Healthcare System, Martinez, CA; University of California Davis, Davis, CA; Department of Veterans Affairs Northern California Healthcare System, Martinez, CA [bold]PURPOSE OF THE STUDY:[/bold][br]The primary objective of this study was to evaluate vitamin D treatment regimens and their dose response relationship in repleting vitamin D serum levels. Secondary outcomes included evaluation of the frequency of monitoring vitamin D serum levels and prescription adherence.[br][bold]METHODS USED:[/bold][br]This was a multicenter, retrospective electronic data extraction and analysis in patients who initiated monotherapy with cholecalciferol or ergocalciferol between January 1, 2005 to December 31, 2010. Patients were excluded if they did not have baseline or follow-up labs within 1 year of treatment initiation. Serum 25(OH)D was used as a marker for vitamin D. Ergocalciferol and cholecalciferol groups were separately organized into quartiles by dosing ranges to evaluate the impact on 25(OH)D.[br][bold]SUMMARY OF RESULTS:[/bold][br]A total of 6,412 patients met inclusion criteria for monotherapy treatment with cholecalciferol (n=4,140) and ergocalciferol (n=2,272). Cholecalciferol quartiles I, II, and III had mean daily doses between 600 IU, 900 IU, and 1,100 IU that were associated with an 8, 8.5, and 9.1 ng/mL 25(OH)D increase from baseline, respectively (p=NS between groups). Cholecalciferol quartile IV had a mean daily dose of 2,700 IU, which was associated with a significantly greater increase from baseline of 12.7 ng/mL 25(OH)D when compared to quartiles I, II, and III (p[lt]0.05). Ergocalciferol doses of 6,000 IU, 7,150 IU, and 7,150 IU, representing quartiles I, II, and III were associated with a 16.5, 14.1, and 17 ng/mL 25(OH)D increase from baseline, respectively (p=NS between groups I,II and I,III). Quartile II had a significantly lower 25(OH)D lab change from baseline than quartile III due to having significantly poorer medication adherence (p[lt]0.05). Quartile IV of ergocalciferol had a mean daily dose of 11,000 IU, which was associated with a significantly greater increase from baseline of 19.9ng/ml 25(OH)D when compared to quartiles I, II, and III (p[lt]0.05).[br][bold]CONCLUSIONS:[/bold][br]Lower cholecalciferol and ergocalciferol maintenance doses had similar modest affects on serum vitamin D levels when compared to the highest mean dose used. The dose-response relationship results of the study reinforce current guidelines and literature. Cholecalciferol and ergocalciferol dose-response relationships found in this study may indicate dosing thresholds exist, and with future analysis, may help narrow and optimize vitamin D repletion regimens.[br][br]Nothing to Disclose: SA, GO, SK-M, JM, AS 2012-06-26T11:30:00 332 2012-06-26T00:00:00 1899-12-30T11:30:00 628 381 2754 OR38-2 OR07-01 Tuesday 2328 2012


2324 ENDO12L_OR38-3 ORAL SESSION: Vitamin D: Supplementation, Metabolism [amp] Function (11:15 AM-12:45 PM) The Effect of Weight on Vitamin D Dose Response Andjela T Drincic, Eileen Van Diest, Laura AG Armas, Robert P Heaney, Susan Dowell Universiy of Nebraska Medical Center, Omaha, NE; Creighton University, Omaha, NE Background: Obesity is associated with low 25(OH) vitamin D levels. Recent Endocrine Society Guidelines advise that obese adults need at least 6,000-10,000 IU/d of vitamin D3 to treat and prevent vitamin D deficiency. There are vitamin D dose response studies in normal to overweight populations (1), but none in the frankly obese.[br]Trial Design: The aim of this randomized, single-blind, study was to characterize the dose response of 25(OH)D to 3 different doses of vitamin D3 in a group of obese women and men. Subjects, enrolled during the winter months, were randomly assigned to either 1,000 IU/d, 5,000 IU/d, or 10,000 IU/d of vitamin D3 for 21 weeks. At baseline, 25(OH)D, PTH, calcium, and serum creatinine were measured. Body composition by DXA and skin tone by IMS Smart Probe were also measured. During follow up visits at 1, 3, 6 and 10 weeks, 25(OH)D and calcium were measured. At week 21, serum 25(OH)D, PTH, serum calcium and 2 hour urine calcium and creatinine were obtained.[br]Inclusion/Exclusion Criteria: Healthy women and men with limited sun exposure, ages 19 [ndash] 70 years, BMI [ge] 30.0 kg/m2, low ([lt]1,000 IU/d) intake of vitamin D3, no history of hepatic, renal disease, malabsorptive condition or medications that affect vitamin D metabolism.[br]Results: 62 subjects of Caucasian or Hispanic origin (25 men and 37 women) were enrolled during winter months over two consecutive years (2009 and 2010). Average age was 45.8 years (SD12.5), average BMI 36.9 kg/m2 (SD 5.7). Baseline 25(OH)D levels were as follows: 20.3 ng/ml in the 1,000 IU/d group, 25.7 ng/ml in the 5,000 IU/d group, and 23.2 ng/ml in the 10,000 IU/d group. Mean increments in 25(OH)D increase were as follows: 12.4 (SD 9.7) ng/ml in the 1,000 IU/d group, 27.8 (SD 10.2) ng/mL in the 5,000 IU/d group, and 48.1(SD 19.6) ng/ml in the 10,000 IU/d group. Compliance with therapy was 97.6 %. There were no adverse events during the study.[br]Conclusion: In obese subjects receiving 5,000 IU/d and 10,000 IU/d of vitamin D3 daily, the 25(OH)D rate of rise was approximately 0.5 ng/ml per 100 IU of vitamin D3 a day compared to reports of 1 ng/ml per 100 IU of vitamin D/d in normal weight people. Obese people need approximately twice the daily dose of vitamin D3 as normal weight people to attain the same increment of 25(OH)D. Everyone in the group that received 5,000 IU dose reached a 25(OH)D level [gt] 30 ng/ml.[br][br](1)Heaney RP et al., Human serum 25-hydroxycholecalciferol response to extended oral dosing with cholecalciferol. Am J Clin Nutr 2003;77:204-210.[br][br]Sources of Research Support: Health Future Foundation (HFF) Faculty Development Program awarded to ATD.[br][br]Nothing to Disclose: ATD, EVD, LAGA, RPH, SD 2012-06-26T11:45:00 332 2012-06-26T00:00:00 1899-12-30T11:45:00 1240 381 2755 OR38-3 OR07-01 Tuesday 2329 2012


2325 ENDO12L_OR38-4 ORAL SESSION: Vitamin D: Supplementation, Metabolism [amp] Function (11:15 AM-12:45 PM) Vitamin D Deficiency Is Prevalent in Morbidly Obese Adolescents Prior to Bariatric Surgery Marisa Censani, Emily M Stein, Sharon Oberfield, Don J McMahon, Shulamit Lerner, Jeffrey Zitsman, Elizabeth Shane, Ilene Fennoy Columbia University Medical Center, New York, NY; Columbia University Medical Center, New York, NY; Columbia University Medical Center, New York, NY Background: Bariatric surgery is a widely used and effective treatment for weight loss in extremely obese adults. Its use in obese adolescents is rapidly increasing with the rise of childhood obesity. Obese adults commonly have vitamin D deficiency, inadequate calcium intake and secondary hyperparathyroidism prior to bariatric surgery. Whether similar metabolic abnormalities exist in morbidly obese adolescents is not known.[br]Objective: To determine the prevalence of vitamin D deficiency in morbidly obese adolescents evaluated for bariatric surgery[br]Methods: A retrospective analysis of preoperative laboratory measures from all adolescent patients (n=236) evaluated for bariatric surgery at Columbia University Medical Center between March 2006 and June 2011.[br]Results: Of 236 patients, 219 patients had 25-hydroxyvitamin D (25OHD) and parathyroid hormone (PTH) levels available for review. The group (76 boys and 143 girls; mean age: 15.9 +/- 1.2 years; 43% Caucasian, 35% Hispanic, 15% African-American) had a mean BMI of 47.5 +/- 8.1 kg/m2. Mean serum 25OHD (20.7 +/- 9.8 ng/ml) was in the insufficient range prior to supplementation, and was inversely associated with BMI (r = -0.28, p [lt]0.0001). 25OHD levels were deficient ([lt]20 ng/ml) in 53% and 8% had severe deficiency ([lt]10 ng/ml) while only 18% of patients were found to be vitamin D replete ([gt]30 ng/ml). Only 11 patients (4.8%) had clear secondary hyperparathyroidism, although PTH levels varied inversely with 25OHD (r= -0.24, p= 0.0003). Findings varied by race, with 82% of African-Americans, 59% of Hispanics, and 37% of Caucasian patients vitamin D deficient. Race was the strongest predictor of 25OHD (p[lt]0.0002). African American race, BMI, and PTH explained 21% of the variance in 25OHD (p [lt] 0.0001).[br]Conclusion: The vast majority of adolescent patients presenting for bariatric surgery have sub-optimal vitamin D levels, with over half in the frankly deficient range. Severely obese adolescents who are African-American and had higher BMIs were at greatest risk for vitamin D deficiency. These results support screening all morbidly obese adolescents for vitamin D deficiency, and repleting those who are deficient. This is particularly important prior to bariatric surgery where weight loss and decreased calcium and vitamin D absorption in some procedures may place these patients at further risk.[br][br]Sources of Research Support: This work is supported by an NIH NIDDK 5T32 DK 06552-07 in Pediatric Endocrinology (PI SE Oberfield).[br][br]Nothing to Disclose: MC, EMS, SO, DJM, SL, JZ, ES, IF 2012-06-26T12:00:00 332 2012-06-26T00:00:00 1899-12-30T12:00:00 1671 381 2756 OR38-4 OR07-01 Tuesday 2330 2012


2326 ENDO12L_OR38-5 ORAL SESSION: Vitamin D: Supplementation, Metabolism [amp] Function (11:15 AM-12:45 PM) Vitamin D Supplementation in Vitamin D Deficient Patients with Primary Hyperparathyroidism: Impact on Calcium and Parathyroid Hormone Levels Dima Abdelmannan, Kathryn Leciejewski, Heitham Ajlouni, Kristina Pascuzzi, Baha M Arafah Cleveland VA Medical Center, Cleveland, OH; Case Western Reserve University, Cleveland, OH [bold][underline]Background:[/underline][/bold][br]Although vitamin D (Vit-D) deficiency is reported to be common in patients with primary hyperparathyroidism, repletion of this deficiency continues to be a challenge because of the concern for worsening hypercalcemia. There are no current guidelines on the safety of vitamin D repletion or the doses to be used in this setting.[br][bold][underline]Objective:[/underline][/bold][br]To determine the impact of Vit-D repletion in a well characterized group of patients with primary hyperparathyroidism who also have Vit-D deficiency on serum calcium (Ca) and Plasma Parathyroid (PTH) levels.[br][bold][underline]Methods:[/underline][/bold][br]The records of 32 patients with Primary Hyperparathyroidism and Vit-D deficiency managed at the Louis Stokes VAMC were reviewed. All patients had elevated PTH levels ([gt] 65 pg/dL), low serum Vit-D levels ([lt] 30 ng/dL), normal serum creatinine ([lt] 1.5 mg/dL) and an elevated albumin-corrected serum Ca ([gt] 10.1 mg/dL). Patients were given ergocalciferol or cholecalciferol at an average daily dose of 3630 units per day. Repeat measurements of plasma PTH, serum levels of Ca (corrected for albumin), Vit-D, and creatinine were made at 3, 6, and 12 months.[br][bold][underline]Results:[/underline][/bold][br]There were 10 females and 22 male patients with a mean age ([plusmn]SD) of 68.9 [plusmn] 12.9 years, and a mean BMD T score of the femur and spine of -0.1 [plusmn] 1.3 and -0.48 [plusmn] 2.5 respectively in 13 patients. Vit-D levels increased significantly at all intervals from a pretreatment mean level of 22.49 [plusmn] 8.41 to 34.4 [plusmn] 9.09 ng/dL after treatment (P [lt] 0.001). Pretreatment corrected serum Calcium levels were elevated (10.73 [plusmn] 0.33 mg/dL) and did not change 3, 6, or 12 months after Vit D replacement (10.8 [plusmn] 0.5, 10.59 [plusmn] 0.47, 10.68 [plusmn] 0.3 mg/dL, respectively; P = 0.89). However, despite the fact that the corrected Ca level remained virtually unchanged, plasma PTH decreased significantly over the treatment period from 123.7 [plusmn] 69.55 in the pretreatment group to 102.5 [plusmn] 42.38 pg/dL after treatment (P = 0.03).[br][bold][underline]Conclusion:[/underline][/bold][br]Vitamin D repletion in vitamin D deficient patients with Primary hyperparathyroidism and hypercalcemia can be safely done under close monitoring. In these patients, the dose required to raise serum vitamin D concentrations to physiological levels remains unknown, although our data suggest that it may not be different from others without primary hyperparathyroidism. By eliminating the secondary cause of hyperparathyroidism, Vit-D treatment in this setting reduces the PTH levels significantly without worsening the hypercalcemia.[br][br]Nothing to Disclose: DA, KL, HA, KP, BMA 2012-06-26T12:15:00 332 2012-06-26T00:00:00 1899-12-30T12:15:00 642 381 2757 OR38-5 OR07-01 Tuesday 2331 2012


2327 ENDO12L_OR38-6 ORAL SESSION: Vitamin D: Supplementation, Metabolism [amp] Function (11:15 AM-12:45 PM) The Role of the Vitamin D Receptor in Innate Immunity; a Study in HVDRR Patients Dov Tiosano, Gizi Wildbaum, Oleg Verbitsky, Vardit Gepstein, Yosef Weisman, Nathan Karin, Amos Eztioni Meyer Children[apos]s Hospital, Rambam Health Care Campus, Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel; Rappaport Family Institute for Research in the Medical Sciences, Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel; Meyer Children[apos]s Hospital, Rambam Health Care Campus, Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel; Technion, Israel Institute of Technology, Haifa, Israel; SackleSchool of Medicine, Tel Aviv University, Tel Aviv, Israel Background: Recently it has been shown that 1,25(OH)[underline]2[/underline]D[underline]3[/underline] through its receptor(VDR)has a major role in innate immunity. Activation of TLR up regulates VDR and Cyp27B1 genes expression leading to upregulate the intracellular killing pathway by enhancing cathelicidin expression. Hereditary 1,25(OH)[underline]2[/underline]D[underline]3[/underline] resistant rickets(HVDRR)is caused by mutations in the VDR gene. Surprisingly, HVDRR patients(Pts)do not present a higher tendency for infectious or autoimmune diseases. Objectives:The aim of the study was to investigate the role of VDR in innate immunity. Methods:15 Pts and 17 controls(C) were investigated. Monocytes were incubated with or without 25OHD[underline]3[/underline] and were stimulated by LPS. TLR2, cathelicidin, DEFB4 and factors that affect cathelicidin expression C/EBP[epsilon],C/EBP[beta] as well as the VDR and the VD response element-BP(hnRNP) expression were analyzed by RT-PCR. Cytokines were measured after stimulation with anti CD3/CD28 and [alpha]GalCer. After incubation with 25OHD[underline]3[/underline] 1,25(OH)[underline]2[/underline]D[underline]3[/underline] levels were similar in Pts and C. Results: VDR, hnRNP, cathelicidin,C/EBP[epsilon],C/EBP[beta] and TLR2 expression did not change in Pts after incubation with 25OHD[underline]3[/underline], whereas in C cathelicidin increased from 1.5[plusmn]0.2 to 3.5[plusmn]0.5,p[lt]4E-9, C/EBP[epsilon] from 1.2[plusmn]0.13 to 2.4[plusmn]0.2,p[lt]2E-9,C/EBP[beta]1.2[plusmn]0.1 to 2.1[plusmn]0.2,p[lt]2E-9, VDR from 1.3[plusmn]0.1 to 2.6[plusmn]0.35,p[lt]6E-9 and hnRNP 1.3[plusmn]0.13to 1.7[plusmn]0.14,p[lt]6E-9.TLR2 declined in C from 1.5[plusmn]0.1to 1.1[plusmn]0.1,p[lt]9E-7. Before incubation with 25OHD[underline]3[/underline] TNF[alpha]and IL17 were elevated in Pts as compare to C, 228[plusmn]45 vs. 28[plusmn]7pg/ml,p[lt]0.005 and 296[plusmn]5 vs. 227[plusmn]42 pg/ml,p[lt]0.005,resp.IL4,IL10 were similar. After incubation with 25OHD3 no changes were found in IL4, IL10,TNF[alpha]and IL17 expression in Pts ,whereas in C IL10 increased from 437[plusmn]65 to 716[plusmn]47pg/ml,p[lt]6E-8,IL4 from 85[plusmn]13 to 200[plusmn]38pg/ml,p[lt]0.004, TNF[alpha] from 27[plusmn]4 to 40[plusmn]3pg/ml,p[lt]5E-7 and IL17 declined from 228[plusmn]33 to 75[plusmn]9pg/ml,p[lt]1E-9. DEFB4 and [alpha]GalCer-stimulated INF[gamma] expression were similar in Pts and C before and after 25OHD[underline]3[/underline]. Conclusions: Evidence from HVDRR pts reveal that 1,25(OH)[underline]2[/underline]D[underline]3[/underline] through the VDR has a major role in enhancing cathelicidin, C/EBP[epsilon],C/EBP[beta] expression as well as IL4 and IL10 and suppressing TNF[alpha]and IL17 expression. Furthermore, 1,25(OH)[underline]2[/underline]D[underline]3[/underline] through the VDR attenuates the intracellular responses following TRL2 activation by suppressing TRL2 and enhancing hnRNP expression. Yet, other VDR-independent mechanisms may be involved in adapting the innate and adaptive immunity in HVDRR, protecting the patients from severe infections.[br][br]Nothing to Disclose: DT, GW, OV, VG, YW, NK, AE 2012-06-26T12:30:00 332 2012-06-26T00:00:00 1899-12-30T12:30:00 2144 381 2758 OR38-6 OR07-01 Tuesday 2332 2012


2328 ENDO12L_OR39-1 ORAL SESSION: Health Outcomes, Professional Education [amp] Health Care Delivery Research (11:15 AM-12:45 PM) Improving Patient Outcomes in Type 2 Diabetes Using Effective Clinician Education Howard BA Baum, Sara C Miller, Stephanie A Stowell, Carolyn A Berry, Bianca R Perri University of Texas Southwestern Medical Center, Dallas, TX; Dallas Diabetes [amp] Endocrine Center, PA, Dallas, TX; Med-IQ, LLC, Baltimore, MD; New York University School of Medicine, New York, NY; , Astoria, NY [bold]BACKGROUND: [/bold]Government policymakers and private medical insurance payors are beginning to implement performance-based physician reimbursement programs for several prevalent disease states, including diabetes. Correspondingly, the AMA has a 3-stage performance improvement continuing medical education model (PI-CME). Med-IQ, an accredited provider of CME, partnered with the Endocrine Society to develop an AMA-style PI-CME program for care of type 2 diabetes in 2008. To date, 292 clinicians have completed the program; significant improvements in performance have been published (1). Changes in patient outcomes, however, have not been demonstrated. Here we report a study evaluating the impact on patient health of completion of PI-CME, the impact of partial completion, and the difference when clinicians participate in less intensive CME endeavors.[br][bold]METHODS: [/bold]Linked chart-review data was used to evaluate clinical indicators of patient health (A1C, lipids, BP) before and after clinician participation in PI-CME and traditional CME activities. Mean values were compared using [italic]t-tests[/italic] and repeated-measures ANOVA was conducted to assess the effect of the type of education.[br][bold]RESULTS: [/bold]Results showed significant changes in measures of health in 323 patients from 44 clinicians who completed the entire PI process. Significant improvements in mean A1C (8.42 to 7.46%, [italic]P[/italic] [lt] 0.001), LDL (111 to 94 mg/dL, [italic]P[/italic] [lt] 0.001), HDL (45 to 47 mg/dL,[italic] P[/italic] [lt] 0.001), and percent of patients achieving BP goals (20 to 40%, [italic]P[/italic] [lt] 0.001) were seen. Improvements in each value were also seen in 227 patients from 27 clinicians who participated in less intensive CME. Hemoglobin A1C values decreased (8.1 to 7.7%), LDL decreased (102 to 91 mg/dL), HDL increased (44 to 46 mg/dL) and the % patients with BP [lt] 130/80 increased (27 to 34 %). There was a significant effect by clinician participant group for A1C ([italic]P[/italic] = 0.01) and LDL ([italic]P = [/italic]0.04), indicating that health improvements were greater in those cared for by PI completers. Improvements in each of the measures were seen in 68 patients cared for by 9 clinicians who participated in most, but not all, of the PI program; magnitude of improvements were not statistically significantly different from the completer group (A1C ([italic]P[/italic] = 0.55), LDL-C ([italic]P[/italic] = 0.22), or HDL ([italic]P[/italic] = 0.08)).[br][bold]CONCLUSIONS: [/bold]We believe that this is the first study to show improved patient outcomes which can be attributed to clinician participation in a performance improvement education program in type 2 diabetes.[br][br](1) Stowell SA, Karcher RB, Bartel RC, Berry CA, King L, Carter RD, Cornish J, Mencia WM. Results of a Performance Improvement Initiative in Diabetes Care. CE Measure. 2010;4:34-39.[br][br]Sources of Research Support: Financial support for data collection and analysis through an unrestricted educational grant from sanofi awarded to Med-IQ. Funding organization played no role in collection, analysis, or presentation of data.[br][br]Nothing to Disclose: HBAB, SCM, SAS, CAB, BRP 2012-06-26T11:15:00 360 2012-06-26T00:00:00 1899-12-30T11:15:00 1776 382 2759 OR39-1 OR15-01 Tuesday 2333 2012


2329 ENDO12L_OR39-2 ORAL SESSION: Health Outcomes, Professional Education [amp] Health Care Delivery Research (11:15 AM-12:45 PM) The Impact of Awareness Activities on the Appropriateness of Pharmacologic Treatment in Type 2 DM Patients in Turkey Ilhan Satman, Sazi Imamoglu, Candeger Yilmaz, ADMIRE Study Group Istanbul University, Istanbul Faculty of Medicine, Istanbul, Turkey; Uludag University, Faculty of Medicine, Bursa, Turkey; Ege University, Faculty of Medicine, Izmir, Turkey [bold]Introduction: [/bold]Clinical practice guidelines on diabetes mellitus (DM) was created by The Society of Endocrinology [amp] Metabolism, Turkey (SEMT). The ADMIRE Project is designed to evaluate the effect of implementation activities to increase physicians[apos] awareness on guidelines.[br][bold]Methods:[/bold] A total of 180 physicians from national representative diabetes clinics kept medical records of 885 T2DM patients for 6 months, without any interference to their routine practice. Then, following [ldquo]awareness activities[rdquo] they kept medical records of 1616 T2DM patients for 6 months. Adherence to guidelines was scored to reach a total adherence to follow-up procedures for medical history, physical examination and laboratory evaluation. Here, results from [ldquo]before[rdquo] and [ldquo]after[rdquo] implementation of awareness program are compared.[br][bold]Results: [/bold]Data before and after the implementation of awareness program were similar (age 55 vs 57 yrs, women 62% vs 59% and DM duration 7.1 vs 7.4 yrs). Treatment preference in before and after implementation of awareness program were OAD: 46% vs 55%, insulin: 11% in both, OAD+insulin: 25% vs 26%. The proportion of patients who self-monitored blood glucose increased by 11% for both baseline (before 46% vs after 57%) and follow-up (before 61% vs after 72%) after implementation of [quot]awareness program[quot] (p[gt]0.001). The proportion of patients who self-monitored blood glucose increased by 15% at follow-up as compared to baseline visit, during both before (baseline 46% vs follow-up 61%) and after (baseline 57% vs follow-up 72%) implementation of [quot]awareness program[quot]. Overall [italic](overtreatment + undertreatment)[/italic] non-adherence rate to antidiabetic treatment recommendations, decreased from 40.0% to 32.5% after implementation of [quot]awareness program[quot] [italic](p[lt]0.001)[/italic]. Undertreatment rate decreased from 35.7% to 22.2% after implementation of [quot]awareness program[quot]. Overtreatment rate increased from 4.2% to 10.3% after implementation of [quot]awareness program[quot]. Non-adherence rate to antihypertensive treatment recommendations, decreased from 21.4% to 12.7% after implementation of [quot]awareness program[quot] [italic](p[lt]0.001)[/italic]. Non-adherence rate to antilipid treatment recommendations, decreased from 13.6% to 9.2% after implementation of [quot]awareness program[quot] [italic](p=0.002)[/italic].[br][bold]Conclusion:[/bold] Implementation of awareness program increased the adherence rate of physicians to SEMT guidelines in terms of antidiabetic, antihypertensive and antilipid treatment.[br][br]Nothing to Disclose: IS, SI, CY 2012-06-26T11:30:00 360 2012-06-26T00:00:00 1899-12-30T11:30:00 1949 382 2760 OR39-2 OR15-01 Tuesday 2334 2012


2330 ENDO12L_OR39-3 ORAL SESSION: Health Outcomes, Professional Education [amp] Health Care Delivery Research (11:15 AM-12:45 PM) Qualitative Outcomes of a Diabetes Simulation for Internal Medicine Residents Diane Donegan, Rose Mader, Sara Weigel, Kurt Kennel Mayo Clinic, Rochester, MN; Mayo Clinic, Rochester, MN Aim: To assess the impact of a simulation of type 1 diabetes mellitus on physician trainee knowledge and attitudes.[br]Method: From 2007 to 2011, 264 internal medicine residents were invited to participate in a three-day immersive simulation of life as a patient with newly diagnosed type 1 diabetes. Following an education session with a diabetes nurse educator and dietician, residents were instructed to perform self blood glucose monitoring 4 times a day; self administer 0.9% saline in lieu of insulin, modify and record their diet and physical activity for three days. Residents participating between 2008 and 2011 were requested to post a [apos]blog[apos] entry reflecting on their experience in preparation for a facilitated group debriefing session.[br]Results: Sixty-two blog entries were available for qualitative assessment. Key themes from each blog entry were abstracted, grouped when similar and, if applicable, categorized under one of six ACGME core competencies. Many (28) entries documented learner enthusiasm for the simulation as a component of their diabetes curriculum. Even so, most simulation participants noted performance challenges including meal choices/planning (30), fear or pain of fingersticks/injections (20), or acknowledged frustration or poor adherence (13). A majority of participants commented on psychosocial impact of diabetes including embarrassment (17) and conflicts with daily routine (19) which was consistent with the frequency of entries indicating increased empathy (29) desire to improve communication and counseling skills (22). The importance of diabetes education and resources within the systems of care was noted less frequently (12). Reported themes spanned all six ACGME core competencies but predominately associated with interpersonal and communication skills.[br]Conclusion: Despite performance challenges, a diabetes simulation with residents as a learner-patient is well received and improves knowledge and attitudes regarding persons with diabetes mellitus.[br][br]Nothing to Disclose: DD, RM, SW, KK 2012-06-26T11:45:00 360 2012-06-26T00:00:00 1899-12-30T11:45:00 1717 382 2761 OR39-3 OR15-01 Tuesday 2335 2012


2331 ENDO12L_OR39-4 ORAL SESSION: Health Outcomes, Professional Education [amp] Health Care Delivery Research (11:15 AM-12:45 PM) A Simple Curriculum Change Increased Medical Student Comfort with Transgender Medicine Joshua D Safer Boston University School of Medicine, Boston, MA Introduction: The biggest barrier to safe therapy for transgender patients is the lack of access to care. Because transgender medicine is not taught in most medical curricula, too few physicians are comfortable with the treatment of transgender conditions. Transgender patients, already stigmatized by societal discomfort with sexual issues, are forced to suffer additional risk and seek treatment from dubious sources.[br]To begin to increase knowledge of transgender medicine by physicians, transgender treatment could become part of standard medical curricula.[br]The hypothesis for this study was that a simple curriculum in medical school that taught rigidity of gender identity, classic treatment regimens, and monitoring requirements of treatment regimens would significantly increase students[apos] willingness to care for transgender patients.[br]Methods: A questionnaire was administered wherein students were asked their predicted comfort using hormones to treat transgender individuals. Students were asked to imagine that they were primary care providers seeing a patient requesting transgender care for the first time. The questionnaire was administered anonymously through an on-line program.[br]The course was part of the mandatory 2nd-year curriculum. The questionnaire was administered a month prior to the course and a month after the course. The shifts in views of the 2nd-year students were contrasted with the views of students in the other years of medical school.[br]Results: Prior to the course, 31% of students self-reported anticipated discomfort with caring for transgender patients. In addition, 3% of students reported the opinion that the treatment was not a part of conventional medicine. Students in the 2nd-year class were no different than students in the other years of school.[br]When queried subsequent to the course, the 2nd-year students reported a dramatic 59% drop in discomfort with transgender care (p[lt]0.005) and no 2nd year students reported the opinion that treatment was not a part of conventional medicine (p=0.01).[br]For students in the other years of medical school there was a notable but more modest 20% drop in discomfort (perhaps due to the impact of the study administration) but no statistical change in the percentage believing the care was not part of conventional medicine.[br]Conclusion: A simple curricular change in medical school significantly increased students[apos] self-reported willingness to care for transgender patients, a hopeful omen for future access to care.[br][br]Nothing to Disclose: JDS 2012-06-26T12:00:00 360 2012-06-26T00:00:00 1899-12-30T12:00:00 138 382 2762 OR39-4 OR15-01 Tuesday 2336 2012


2332 ENDO12L_OR39-5 ORAL SESSION: Health Outcomes, Professional Education [amp] Health Care Delivery Research (11:15 AM-12:45 PM) Subspecialty Rotation in Medicine Residency Program Is Associated with Significant Diabetes Knowledge Gain Aarti Manchanda, Ambika Babu, Alan Schwartz, Leon Fogelfeld John H Stroger Jr Hospital of Cook County, Chicago, IL; University of illinois, Chicago, IL; John H Stroger Jr Hospital of Cook County, Chicago, IL [bold]Introduction: [/bold]The impact of an elective rotation in endocrinology/diabetes during medicine residencies has not been studied. The aim was to assess the change in knowledge of diabetes management among trainees before and after such rotation.[br][bold]Methods: [/bold]A study group (184 medical students and 1[sup]st[/sup] to 3[sup]rd[/sup] year residents) were given a pretest of 15 multiple choice questions about various aspects of diabetes at the start of the rotation. This pretest was designed to be [ldquo]easy[rdquo] as an encouraging warm-up. At the end of the rotation, 4 weeks later, a more [ldquo]difficult[rdquo] post-test was given which was matched for the areas covered. For validation purposes, 14 trainees did both the [ldquo]easy[rdquo] pretest and [ldquo]difficult[rdquo] posttest at the beginning and end of the rotation. A control group of 12 trainees, who did not have the rotation, also took both the tests at the beginning and at the end. All the results are expressed as mean % correct scores.[br][bold]Results: [/bold]In the validation group, the scores at start were lower for the [ldquo]difficult[rdquo] than for the [ldquo]easy[rdquo] tests (61.9 vs. 45.7,p=0.002) showing the differences in difficulty between them. The improvement, at the end of the rotation was significant for both [ldquo]easy[rdquo] and [ldquo]difficult[rdquo] tests (76.1 and 70.4). The control group did not show any improvement at end of 4 weeks in either the [ldquo]easy[rdquo] or [ldquo]difficult[rdquo] tests (p=0.55 vs.0.39). The 2 way ANOVA test also showed that at the start, the knowledge was similar between the validation and control group using either the [ldquo]easy[rdquo] or [ldquo]difficult[rdquo] test. However, the scores were much higher in the validation than control group for both easy and difficult posttests (76.1 vs 56.6, p=0.001 and 70.5 vs 43.9, p[lt] 0.001). In the study group, there was an improvement in the scores at end of the rotation ([ldquo]Easy[rdquo] pretest vs. [ldquo]difficult[rdquo] posttest, 63.4 and 66.8, p[lt]0.014). Considering the difference between the 2 tests, the increase in knowledge for the study group is even bigger and similar to the one in the validation group. There were no significant differences in the scores at baseline and at end across the different levels of trainees.[br][bold]Conclusions: [/bold]Clinical endocrine rotation with an emphasis on diabetes management was associated with a significant gain of knowledge. A structured rotation with focused diabetes education may be needed not as an elective but as an integral component of medicine residency programs. Further studies to evaluate whether such training will translate into a better patients outcome are needed.[br][br]Nothing to Disclose: AM, AB, AS, LF 2012-06-26T12:15:00 360 2012-06-26T00:00:00 1899-12-30T12:15:00 703 382 2763 OR39-5 OR15-01 Tuesday 2337 2012


2333 ENDO12L_OR39-6 ORAL SESSION: Health Outcomes, Professional Education [amp] Health Care Delivery Research (11:15 AM-12:45 PM) Predictors of Prolonged Care for Transition-Aged Patients in a Tertiary Pediatric Endocrinology Clinic Matthew D Wise, Christine Yu, Tamara J Vokes University of Chicago, Chicago, IL; University of Chicago, Chicago, IL Background: The transition from pediatric to adult care for adolescents with chronic endocrinopathies is recognized by patients and providers as inadequate, correlates with outcomes, and has emerged as a targeted focus for systematic improvement. There is limited data to characterize the transition practices of pediatric endocrinologists.[br]Objective: To determine the prevalence of transition-aged ([ge]18 years) patients in a tertiary pediatric endocrinology clinic and identify clinical variables associated with delayed transition to adult care.[br]Methods: All patient visits to the pediatric endocrinology clinic (jointly located with the adult endocrinology clinic) at the University of Chicago from a 5-year period were analyzed. Delayed transition was defined as age [ge] 18 at the most recent clinic encounter. ICD-9 codes were used to define primary diagnosis. The associations between delayed transition and clinical predictors (diagnosis, provider, age, gender, race, and ethnicity and insurance coverage) were analyzed using chi-square test and multivariate logistic regression.[br]Results: The study included 2296 patients (55% female) seen by pediatric endocrinologists over 5 years. There were 166 (7.2%) patients [ge] 18 years old at the most recent encounter. Duration of care (OR 1.5 per year of care, 95% CI 1.4-1.7, p[lt]0.001), receiving care from one specific provider (OR 2.2, 95% CI 1.6-3.0, p[lt]0.001), and female gender (OR 1.8, 95% CI 1.3-2.5, p=0.001) were associated with delayed transition. Diagnoses associated with delayed transition, when analyzed by multivariate regression and controlling for the variables above, included diabetes (OR 2.6, 95% CI 1.7-3.9, p[lt]0.001), pituitary (OR 2.6, 95% CI 1.5-4.5, p=0.001), neoplastic (OR 4.4, 95% CI 1.7-11.4, p=0.002), and chromosomal-related disorders (OR 2.8, 95% CI 1.1-7.3, p=0.04). Thyroid, adrenal, growth, and pubertal disorders were not associated with delays in transition. Race/ethnicity and insurance status (commercial vs. public) did not predict delay in transition.[br]Conclusions: Even in a combined adult and pediatric clinic setting at an academic center, one in fourteen pediatric endocrinology patients will not be transitioned to an adult endocrinologist by 18 years. Female gender, individual provider practice style, and diagnoses that require intensive chronic management appear to impede the transition of care.[br][br]Nothing to Disclose: MDW, CY, TJV 2012-06-26T12:30:00 360 2012-06-26T00:00:00 1899-12-30T12:30:00 617 382 2764 OR39-6 OR15-01 Tuesday 2338 2012


2334 ENDO12L_OR40-1 ORAL SESSION: Management of Diabetes (11:15 AM-12:45 PM) Remission of Type 2 Diabetes Following Ileal Interposition with Sleeve Gastrectomy/Diverted Sleeve Gastrectomy Sunil K Kota, Surendra Ugale, Neeraj Gupta, Vishwas G Naik, Kirtikumar D Modi Medwin Hospital, Hyderabad, India; Kirloskar Hospital, Hyderabad, India Objective:We evaluated the efficacy of ileal interposition(II)+sleeve gastrectomy(SG)/diverted SG(DSG) for control of type 2 diabetes (T2DM) and metabolic abnormalities.[br]Methods:II+SG was performed on 43 patients (M:F=25:18).The inclusion criteria:T2DM [gt]1 year duration,age 25- 70 years,stable weight for 3 months and stimulated C-peptide level [gt]1.5 ng/ml.17 patients (M:F=12:5) underwent II+DSG,based on adverse profile like longer duration of diabetes,lower BMI,poorer C-peptide response.The primary outcome was remission of diabetes (HbA1C[lt]6.5% without OHAs/Insulin) and secondary outcomes were reduction in OHA requirement and improvement in metabolic profile.Patients were followed up at 3 monthly intervals.[br]Results:Patients subjected to II+SG had mean age 47.2[plusmn]8.2years, DM duration 10.1[plusmn]9.2 years and BMI 33.2[plusmn]7.8 kg/m2.All patients had poorly controlled T2DM (HbA1C-9.6[plusmn]2.1%).30(70%) patients had hypertension,20(46%) had dyslipidemia and 18(42%) had microalbuminuria.Mean follow up was 20.2[plusmn]8.6 months(range:4-40 months).Postoperatively glycemic parameters improved at all intervals (p[lt]0.05).20 patients(47%) had remission in T2DM and the remaining showed significantly decreased OHA requirement.27 patients(90%) had remission in hypertension.There was a declining trend in lipids and microalbuminuria.Patients with duration of T2DM [lt] 10 years, stimulated C-peptide [gt]4 ng/ml and BMI [gt] 27 kg/m2 performed better.[br]Patients subjected to II+DSG, had mean age 50.7[plusmn]8.1 years,DM duration 15.1[plusmn]5.8 years and BMI 29.2[plusmn]7.5 kg/m2.8(45%) patients had hypertension,7(39%) had dyslipidemia and 7(39%) had microalbuminuria.Mean follow-up data was 9.1[plusmn]5.3 months(range: 3[ndash]21 months).12(70.5%)patients had diabetes remission, and the remaining five showed significantly decreased OHA requirement.7 patients (87.5%) had remission in hypertension.Significant decline was observed in the glycemic, lipid parameters and microalbuminuria at all intervals (p[lt] 0.05).HbA1C reduction was higher than decline in BMI, justifying weight loss independent glycemic benefits.[br]Discussion:The surgery addresses the foregut and hindgut mechanisms for DM control.The SG component restricts calorie intake and induces ghrelin loss.II leads to rapid stimulation of interposed ileal segment by ingested food leads resulting in augmented GLP-1 secretion.Improvement in hypertension, lipid profile and microalbuminuria justify its metabolic benefits.[br]Conclusion:II+SG/DSG can control Type 2 DM and associated metabolic abnormalities.[br][br]Nothing to Disclose: SKK, SU, NG, VGN, KDM 2012-06-26T11:15:00 Theater C 2012-06-26T00:00:00 1899-12-30T11:15:00 1048 383 2765 OR40-1 OR11-02 Tuesday 2339 2012


2335 ENDO12L_OR40-2 ORAL SESSION: Management of Diabetes (11:15 AM-12:45 PM) Effectiveness of Glucagon Solution during 8 Hours of Aging Parkash A Bakhtiani, Joseph El Youssef, Jessica R Castle, W Kenneth Ward, Deborah L Branigan, Nicholas Caputo, Tara Stonex, Melanie A Jackson Oregon Health and Science University, Portland, OR; Legacy Health System (Research), Portland, OR [bold]INTRODUCTION:[/bold] Due to glucagon[apos]s chemical instability, commercial preparations must be used immediately after reconstitution(1). As part of a clinical test of a artificial endocrine pancreas(2,3), we used glucagon and insulin for closed loop control of type 1 diabetes in 28 studies, each of which lasted 33 h. Glucagon (GlucaGen, Novo) was given on 69 occasions over the 900 hours(4). The current research was undertaken to measure kinetics of chemical and clinical loss of glucagon purity and efficacy.[br][bold]METHODS:[/bold] To address clinical efficacy, we asked whether freshly reconstituted glucagon was superior to glucagon aged for up to 8 hours after being reconstituted.[br]To assess for chemical aggregation and degradation, we carried out reversed phase high-performance liquid chromatography (rpHPLC) and two assays for amyloid fibril formation on glucagon formulated in acidic or alkaline buffers.[br][bold]RESULTS:[/bold] Glucagon was successful in avoiding hypoglycemia ([lt] 70 mg/dl) in 56 out of 69 occasions (81%). We found no correlation (R[sup2] = 0.018) between duration of aging up to 8 hours and effectiveness of glucagon doses in the closed loop setting. With glucagon prepared in acidic buffers, there was little protein degradation using rpHPLC, but it rapidly formed amyloid fibrils demonstrated both by Thioflavin T binding and intrinsic tryptophan fluorescence. Fibril formation was maximal at 48 hours after the onset of aging at 37 C. Our previously-published results found that glucagon buffered with glycine at pH 10 showed very little loss of the monomeric form by size-exclusion HPLC in contrast to the cytotoxicity and amyloid found after aging of the acidic glucagon(5).[br][bold]CONCLUSIONS:[/bold] Though in approximately 80% of cases, small doses of subcutaneous glucagon in bihormonal closed loop experiments are effective in preventing hypoglycemia, the cause of the 20% failure rate is unclear. We saw no evidence of loss of clinical activity after 8 h of aging. However, there is rapid development of amyloid fibrils when glucagon is aged at acid pH. The instability of acidic preparations of glucagon is primarily due to amyloid formation, rather than protein degradation. Reconstitution of commercially-available glucagon every 8 h appears to be effective in a research setting, but for outpatient closed loop use of glucagon, more stable formulations must be developed.[br][br]1. Onoue S et al., Pharmaceutical Research 2004; 21:7. 2. Peter GJ et al., Conf Proc IEEE Eng Med Biol Soc. 2011 397-400. 3. Ward WK et al., J Diabetes Sci Technol. 2011 1;5(6):1373-80. 4. El Youssef et al., J Diabetes Sci Technol. 2011 Nov 1;5(6):1312-26. 5. Ward WK et al., J Diabetes Sci Technol 2010; 4:6.[br][br]Nothing to Disclose: PAB, JEY, JRC, WKW, DLB, NC, TS, MAJ 2012-06-26T11:30:00 Theater C 2012-06-26T00:00:00 1899-12-30T11:30:00 1828 383 2766 OR40-2 OR11-02 Tuesday 2340 2012


2336 ENDO12L_OR40-3 ORAL SESSION: Management of Diabetes (11:15 AM-12:45 PM) Long-Term Neurocognitive Development of Critically Ill Children 4 Years after Randomization to Insulin-Titrated Age-Adjusted Normoglycemia or to Usual Care Dieter Mesotten, Marijke Gielen, Caroline Sterken, Dirk Vlasselaers, Greet Hermans, Ilse Vanhorebeek, Pieter J Wouters, Greet Van den Berghe Catholic University of Leuven (KU Leuven), Leuven, Belgium A large prospective randomized controlled trial performed in a tertiary referral pediatric ICU revealed that targeting age-adjusted normoglycemia (TGC) reduced ICU morbidity and mortality, but increased the risk of hypoglycemia [lt]40mg/dl (25% of patients) (1). As both hyper- and hypoglycemia may adversely affect the developing brain of young children, assessing long-term neurocognitive development was required to validate the short-term benefit.[br]We here report on the pre-planned follow-up study of all 700 patients originally included in the trial at a median (IQR) age of 1.3(0.3-4.9)y. For comparison 216 controls, unrelated children and patients[apos] siblings, were examined. Neurodevelopmental testing encompassed clinical neurologic examination and age-adjusted tests for intelligence (Wechsler IQ scales), memory (Children[apos]s Memory Scale), visual-motor integration (VMI-Beery Buktenica Developmental Test), attention and executive functions (ANT-Amsterdam Neuropsychological Tasks) and behavior (Child Behavior Check List).[br]At 3.9(3.8-4.0)y after inclusion, survival rate remained higher with TGC (risk-adjusted OR 1.89 (95% CI 1.01-3.61) (p=0.04). 17% of TGC and 15% of UC patients declined participation or were lost to follow-up (p=0.6). In 9% of patients, also equally distributed, neurocognitive function tests could not be performed due to severe mental/psychomotor impairment. As compared with healthy control children, more neurological abnormalities were present in patients (p[lt]0.001), irrespective of insulin treatment. Full scale IQ was lower in patients [88(74-99)] than controls [103(91-111)] (p[lt]0.001) but comparable between TGC [88(74-100)] and UC [89(74-99)] patients (p=0.7). Similar differences were demonstrated for verbal IQ, performance IQ, memory, VMI and behavior. The execution of complex cognitive processes (ANT) was 13-20% better (all p[lt]0.03) in patients who had been treated with TGC during intensive care than in the UC group. Similar differences were present for patients with or without pre-existing syndrome or pathology with potential influence on mental/psychomotor functioning. Also the results were confirmed by imputation for minor differences in baseline demographics between patients and controls.[br]In conclusion, 4 years after randomization, children who had been critically ill scored worse than healthy controls. Despite hypoglycemia, the survival benefit with TGC during intensive care was maintained and neurocognitive development slightly improved.[br][br]1. Vlasselaers D et al.: Lancet 2009; 373: 547-556.[br][br]Sources of Research Support: * Funded by the Methusalem program of the Flemish government (to GVdB), IWT-TBM070695 (to GVdB,DM, IV), PhD doctoral fellowship of the Fund for Scientific Research (FWO) Flanders, Belgium (to MG), Post-doctoral fellowship of the Fund for Scientific Research (FWO) (to DM, GH). * TRIAL REGISTRATION: ClinicalTrials.gov Identifier NCT00214916. * DM, MG and CS share first authorship.[br][br]Nothing to Disclose: DM, MG, CS, DV, GH, IV, PJW, GVdB 2012-06-26T11:45:00 Theater C 2012-06-26T00:00:00 1899-12-30T11:45:00 1125 383 2767 OR40-3 OR11-02 Tuesday 2341 2012


2337 ENDO12L_OR40-4 ORAL SESSION: Management of Diabetes (11:15 AM-12:45 PM) Adherence to Clinical Practice Guidelines among Youth with Type 2 Diabetes in British Columbia, Canada Shazhan Amed, Behrad Besharatian, Kim Reimer, Kimberly Nuernberger, Patrick McCrea, Dieter Ayers, Sema Aydede, Jean-Paul Collet University of British Columbia, Vancouver, Canada; University of British Columbia, Vancouver, Canada; British Columbia Ministry of Health, Victoria, Canada Type 2 diabetes (T2D) is associated with serious long-term complications. Optimal glycemic control in youth diagnosed with T2D is critical in preventing diabetes-related complications. Evidence to date suggests that the quality of health care services for youth with diabetes is sub-optimal. This study describes adherence to clinical practice guidelines (ACPG) in youth with T2D living in British Columbia, Canada. All youth diagnosed with T2D at [lt]20 years of age between 1996-2008 were identified within linked health administrative datasets using a validated diabetes case-finding algorithm and diabetes differentiating algorithm. Incident cases were followed for a variable period of 2-5 years (N=488 individuals; 2111 person-years) and a cross-sectional sample (n=275) from a 12-month period (Apr 1/06-Mar 31/07) was described. Two measures of ACPG were developed using Canadian clinical practice guidelines: (i) an ordinal interval ACPG score ranging from optimal care=4, good care=3, minimal care=2, and poor care=1; and (ii) a categorical variable where individuals with optimal or good care were classified as being [ldquo]at goal.[rdquo] Outcome variables included mean annual ACPG score and proportion of individuals [ldquo]at goal.[rdquo] Descriptive statistics (means, medians, frequencies and tests of significance) were used. Overall, 68.1% of youth with T2D had poor ACPG ([lt]2 T2D related physician visits and hemoglobin A1c (A1c) tests and no screening tests in a 1-yr period). Mean annual ACPG score decreased from year of diagnosis to 4 years post-diagnosis (1.85 versus 1.49; p[lt]0.001) and as youth aged from 10-14 to 20-24 years (1.85 vs. 1.52; p[lt]0.001). Males (N=218) had a higher mean annual ACPG score compared to females (N=270) (1.74 [95% CI: 1.65-1.84] vs 1.59 [95 CI: 1.51-1.67]; p=0.02) however, this difference disappeared in the 20-24 year age group. In the cross-sectional analysis 31% (85/275) were classified as having good or optimal ACPG ([apos]at goal[apos]). Also, 60% (165/275), 36% (99/275), and 67% (185/275) received at least 2 diabetes related physician visits, A1c tests and screening tests for co-morbidity in that year. This study shows ACPG for youth-onset T2D is sub-optimal in our region and worsens as years from diagnosis increases. Youth transitioning into adulthood are a particularly vulnerable population. Our results are relevant to health professionals and policymakers who directly influence health services and are responsible for developing innovative health system change.[br][br]Waitzfelder, B, et al. Adherence to Guidelines for Youths With Diabetes Mellitus. PEDIATRICS. 2011.[br][br]Nothing to Disclose: SA, BB, KR, KN, PM, DA, SA, J-PC 2012-06-26T12:00:00 Theater C 2012-06-26T00:00:00 1899-12-30T12:00:00 2245 383 2768 OR40-4 OR11-02 Tuesday 2342 2012


2338 ENDO12L_OR40-5 ORAL SESSION: Management of Diabetes (11:15 AM-12:45 PM) Sarcopenic Phenotype in Patients with Newly Diagnosed Type 2 Diabetes Hee Kyoung Jeong, Soon Jib Yoo, Sung Rae Kim, Seong Su Lee, Jang Won Son The Catholic University of Korea, Bucheon, Korea Objective [bold][ndash] [/bold]Sarcopenic phenotype is associated with the increased risk of insulin resistance and type 2 diabetes. We investigated the proportion of sarcopenia and sarcopenic obesity (SO) and their clinical characteristics in patients with newly diagnosed type 2 diabetes.[br]Research design and methods[bold] [ndash] [/bold]We conducted a cross-sectional analysis of 133 (75 men, 58 women) patients with newly diagnosed type 2 diabetes. All patients underwent a 75 g oral glucose tolerance test. Sacopenia was assessed by appendicular skeletal muscle mass (ASM)/weight (%) using dual-energy X-ray absorptiometry and obesity was assessed by a visceral fat area using computed tomography .[br]Results [bold][ndash] [/bold]The mean age of the subjects was 47.14 [plusmn] 12.14 years, the mean HbA1c level was 10.59 [plusmn] 1.94 % and the mean body mass index was 25.53 [plusmn] 3.38 kg/m[sup]2[/sup]. ASM/weight (%) was inversely associated with the log-transformed homeostasis model assessment of insulin resistance (HOMA-IR) and HOMA-[beta]. The proportion of obesity, sarcopenia and SO was 23.3%, 16.5% and 35.3%, respectively. Sarcopenic phenotype was more prevalent in women than in men (Sarcopenia 29.3% vs. 6.7% and, SO 44.8% vs. 28%, respectively). In comparison to normal phenotype (neither obese nor sarcopenia), SO was significantly associated with higher degree of HOMA-[beta] and the surrogate markers of insulin resistance (HOMA-IR and insulin sensitivity index composite). Notably, SO subjects were more insulin resistant compared to obese without sarcopenia subjects, although there was no significant difference in HbA1c between two groups.[br]Conclusions [ndash] We found the high proportion of sarcopenia and SO in patients with newly diagnosed type 2 diabetes. Among metabolic phenotype, SO was strongly associated with the degree of insulin resistance and compensated hyperinsulinemia in the early phase type 2 diabetes.[br][br]Nothing to Disclose: HKJ, SJY, SRK, SSL, JS 2012-06-26T12:15:00 Theater C 2012-06-26T00:00:00 1899-12-30T12:15:00 1397 383 2769 OR40-5 OR11-02 Tuesday 2343 2012


2339 ENDO12L_OR40-6 ORAL SESSION: Management of Diabetes (11:15 AM-12:45 PM) Prolactin Receptor Gene Polymorphisms Are Associated with Gestational Diabetes Trang N Le, Sarah H Elsea, Gary L Francis Virginia Commonwealth University, Richmond, VA; Virginia Commonwealth University, Richmond, VA Background: Gestational diabetes mellitus (GDM) results from inadequate compensatory increases in insulin secretion and pancreatic [beta]-cell mass during pregnancy. Recent data suggest that human placental lactogen (HPL) acts via the prolactin receptor (PRLR) on maternal [beta]-cells to mediate increases in [beta]-cell mass during normal pregnancy. Infants of GDM also have an increased risk of developing type 2 diabetes mellitus (T2DM) in adulthood. Consequently, identification of single-nucleotide polymorphisms (SNPs) in the [italic]PRLR[/italic] gene that are associated with GDM could illuminate potential genetic predispositions towards GDM and improve our understanding of how prenatal events program T2DM in adults.[br]Objective: To determine if [italic]PRLR[/italic] SNPs are associated with GDM.[br]Design/Methods: Deidentified DNA samples from Santiago, Chile were obtained from 96 mother-infant dyads affected by GDM and 96 unaffected mother-infant dyads. 10 [italic]PRLR[/italic] SNPs, located in the promoter region, 3[apos]UTR, or introns 1 or 2, are known to be clinically relevant in studies of breast cancer, multiple sclerosis, or autoimmune disease and were studied using real-time PCR with TaqMan SNP Genotyping Assays.[br]Results: Significant associations were identified in 2 of the 10 SNPs analyzed. The frequency of the variant allele of rs10068521 was higher in GDM cases compared to controls (minor allele C present in 20.2% of GDM cases, 9.7% in controls, p = 0.0435) with an increased GDM risk in those carrying the minor allele (OR 2.36; 95% CI, 1.01-5.54). For rs9292578, the minor allele A was more frequently observed in GDM cases than controls (minor allele present in 20.2% of GDM cases, 9.7% in controls, p = 0.0435), resulting in an increased risk for GDM in those carrying the minor allele (OR 2.36; 95% CI, 1.01-5.54). Of note, in both cases and controls, each sample with the minor allele of rs10068521 also contained the minor allele of rs9292578. The frequency of the rs9292579 variant allele approached but did not reach significance. Both GDM-associated SNPs are located in the promoter region of PRLR.[br]Conclusions: These data suggest that polymorphisms of the PRLR promoter region are associated with GDM, perhaps via decreased efficiency of [italic]PRLR[/italic] transcription and subsequent decreased responsiveness to HPL in pregnancy. Additional studies SNPs of the genes [italic]CSH1[/italic] and [italic]CSH2[/italic], which encode HPL, may help to clarify the role of fetal HPL in modulating maternal GDM risk.[br][br]Sources of Research Support: A.D. Williams Fund, Virginia Commonwealth University, and VCU CTSA UL1RR031990. DNA samples were kindly provided by Dr. Roberto Romero, MD, Perinatology Research Branch, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Department of Health and Human Services, Detroit, Michigan.[br][br]Nothing to Disclose: TNL, SHE, GLF 2012-06-26T12:30:00 Theater C 2012-06-26T00:00:00 1899-12-30T12:30:00 1696 383 2770 OR40-6 OR11-02 Tuesday 2344 2012


2340 ENDO12L_OR41-1 ORAL SESSION: Novel Pituitary Tumor Syndromes [amp] Sellar Metastasis (11:15 AM-12:45 PM) A New Syndromic Association: Pituitary Adenomas, Paragangliomas and Pheochromocytomas Paraskevi Xekouki, Panagiotis Mastorakos, Alex Spyridon Mastroyannis, Dimitrios Avgeropoulos, Aristides Lytras, Charalampos Lyssikatos, Monalisa Ferreira Azevedo, Anelia Horvath, Carl Malcoff, Constantine A Stratakis National Institutes of Health (NIH), Bethesda, MD; National Institutes of Health (NIH), Bethesda, MD; Harvard Medical School, Boston, MA; University of Connecticut Health Center, Farmington, CT We recently reported a potentially new syndromic association, that of a GH-secreting pituitary adenoma and pheochromocytoma/paraganglioma (PHEO/PGL)[sup]1[/sup]. Prompted by this observation, we interrogated our International Registry of mostly pediatric sporadic pituitary adenomas, as well as adult and pediatric syndromic pituitary tumors for the occurrence of pituitary adenomas with PHEO/PGL. A total of 161 cases were investigated: 16 were identified as familial/syndromic and 145 as sporadic. Among the syndromic cases 4 were diagnosed with Cushing[apos]s disease and 12 with [italic]GH[/italic]- and PRL-secreting adenomas. Among the sporadic cases 117 were diagnosed with Cushing[apos]s disease, 8 with GH-secreting adenomas, 5 with prolactinomas, 5 with non-secreting tumors, 1 with craniopharyngioma and 9 with hormonal syndromes and no obvious tumor. Out of the 145 sporadic cases 2 presented with the new syndromic association; 1 case had a GH/PRL pituitary adenoma and PHEO/PGL, and the other one an ACTH-producing adenoma and PGLs. In the second case, mutation screening for [italic]SDHB[/italic], [italic]SDHC,[/italic] [italic]SDHD, and SDHAF2[/italic] genes was negative, whereas mutation screening for the first case is pending. None of the syndromic cases had any PHEO/PGL association, and all were due to other mutations including 2 in [italic]AIP[/italic] and 3 in [italic]MEN1[/italic] genes or were due to yet unknown molecular defects. In addition to the two kindreds with pituitary adenomas and PHEO/PGLs and the previously reported family with acromegaly and PHEO/PGLs, we also identified four previously reported polymorphisms, including His50Arg and Gly12Ser in [italic]SDHD[/italic], as well as, 170A[gt]G,p.His57Arg and 163Ser[gt]Pro in [italic]SDHB[/italic] in the sporadic adenoma population. This represents an unexpectedly higher frequency from what has been reported in the general population for these variants; screening of our own control group is ongoing. Both [italic]SDHB[/italic] 163Ser[gt]Pro and [italic]SDHD[/italic] His50Arg have been associated with mitochondrial dysfunction in a subset of [italic]PTEN[/italic] mutation-negative Cowden and Cowden-like syndrome individuals[sup]2[/sup]. We conclude that as we speculated first, when we described the [italic]SDHD[/italic] mutation in connection with a pituitary tumor and PHEO/PGL, there might be an association of these tumors that may constitute a new syndrome. However, it is rare among sporadic pituitary adenomas, although functional SDHx variants may play a role there, too. This discovery may lead to new therapies targeting mitochondrial oxidation in aggressive pituitary tumors.[br][br](1)Xekouki et al. J Clin Endocrinol Metab 2011; [Epub ahead of print]. (2)Ni et al. Am J Hum Genet. 2008;83:261.[br][br]Nothing to Disclose: PX, PM, ASM, DA, AL, CL, MFA, AH, CM, CAS 2012-06-26T11:15:00 351 2012-06-26T00:00:00 1899-12-30T11:15:00 2183 384 2771 OR41-1 OR22-01 Tuesday 2345 2012


2341 ENDO12L_OR41-2 ORAL SESSION: Novel Pituitary Tumor Syndromes [amp] Sellar Metastasis (11:15 AM-12:45 PM) Familial Pituitary Adenoma and Paraganglioma Syndrome [mdash] A Novel Type of Multiple Endocrine Neoplasia Judit Denes, Francesca Swords, Paraskevi Xekouki, Ajith V Kumar, Eammon R Maher, Christopher A Wassif, Naomi Fersht, Joan Grieve, Stephanie E Baldeweg, Constantine A Stratakis, Marta Korbonits Military Hospital, Budapest, Hungary; Norfolk and Norwich University Hospital, Norwich, UK; Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD; Great Ormond Street Hospital, London, UK; University of Birmingham, Birmingham, UK; Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD; University College London Hospitals, London, UK; National Hospital for Neurology and Neurosurgery, London, UK; University College London Hospitals, London, UK; Barts and The London School of Medicine, Queen Mary University of London, London, UK Familial pituitary adenomas may be caused by AIP, MEN1 and PRKAR1A mutations. Familial phaeochromocytomas/paraganglioms (PHAEO/PARA) may be caused by VHL, SDH, RET, NF1, TMEM127, KIF1Bb and MAX mutations. The presentation of these 2 tumour types together has been described in the past: GHRH-secreting PHAEO/PARA can cause acromegaly due to pituitary hyperplasia. However, in other cases, true pituitary adenomas has been described in association with PHAEO/PARA. Recently a family has been published with the coexistence of pituitary and PHAEO/PARA harbouring an SDHD mutation and showing LOH at the SDHD locus in the pituitary adenoma (1), and we have published data previously (2) on a SDHB-mutation family (Family-1) with the coexistence of pituitary adenoma and PHAEO/PARA. We have now performed LOH studies in this and a further family with an SDHC mutation (Family-2).[br]Family-1: A 33y male with headache, visual loss, hypogonadism, bitemporal hemianopia, elevated prolactin, presented due to a macroadenoma with recent infarction. MRI also revealed a carotid paraganglioma. A novel germline missense mutation was identified in SDHB: c.298T[gt]C;p.S100P. His carrier mother had a surgically-treated macroprolactinoma and was also found to have a benign thyroid nodule, and subclinical Cushing[apos]s syndrome secondary to bilateral adrenal hyperplasia. Prospective screening in the proband[apos]s carrier brother revealed a carotid paraganglioma. LOH was found at the SDHB locus in the prolactinoma of the proband.[br]Family-2: A male patient with glomus jugulare tumour was operated at 38y and 45y. At 53y he was diagnosed with a macroprolactinoma, which is treated with cabergoline. His brother had a glomus tumour at 60y. His 55y first cousin had a clinically nonfunctioning pituitary adenoma showing LH/FSH staining. A novel missense SDHC mutation was found in this family: c.380A[gt]G;p.R127H.[br]These data, together with the published SDHD family, suggest that SDH mutations may result in familial pituitary adenomas associated with PHAEO/PARA tumours.[br][br](1) Xekouki et al.,J Clin Endocrinol Metab.,2011. (2) Brahma et al.,Endocr.Abs.,2009.[br][br]Nothing to Disclose: JD, FS, PX, AVK, ERM, CAW, NF, JG, SEB, CAS, MK 2012-06-26T11:30:00 351 2012-06-26T00:00:00 1899-12-30T11:30:00 936 384 2772 OR41-2 OR22-01 Tuesday 2346 2012


2342 ENDO12L_OR41-3 ORAL SESSION: Novel Pituitary Tumor Syndromes [amp] Sellar Metastasis (11:15 AM-12:45 PM) Progression of Acromegalic Arthropathy Despite Long-Term Biochemical Control: A Prospective Follow-Up Study Kim Claessen, Sharita Ramautar, Alberto Pereira, Jan Smit, Ferdinand Roelfsema, Hans Romijn, Herman Kroon, Margreet Kloppenburg, Nienke Biermasz Leiden University Medical Center, Leiden, Netherlands; Leiden University Medical Center, Leiden, Netherlands; Leiden University Medical Center, Leiden, Netherlands [bold][italic]Background[/italic][/bold][br]Arthropathy is an invalidating complication of acromegaly, despite persisting biochemical disease control, which has a high impact on the quality of life. Prognosis and determinants are currently unknown.[br][bold][italic]Objectives[/italic][/bold][br]To investigate the clinical and radiological disease course of arthropathy during 2.6year follow-up and to assess determinants of outcome in patients with long-term controlled acromegaly.[br][bold][italic]Methods[/italic][/bold][br]In a prospective cohort study, 58 patients (mean age 62,women 41%) with controlled acromegaly for a mean of 17.6years were included. 40 patients(69%) were cured by surgery and, if necessary, received additional radiotherapy, 18(31%) patients were controlled by somatostatin(SMS) analogs. Radiographic progression of joint disease was defined as minimal 1-score increase in the Osteoarthritis Research Society International (OARSI) classification on radiographs of hands, knees and hips over 2.6years. Clinical progression was measured by the Australian/Canadian Osteoarthritis Hand Index(AUSCAN) for hand OA, Western Ontario McMaster Universities Osteoarthritis Index(WOMAC) for lower limb OA, structured physical joint examination and several function tests. Potential risk factors for progression were assessed, and the relationship between clinical and radiographic disease course.[br][bold][italic]Results[/italic][/bold][br]Radiographic progression was observed in more than 70% of patients. Parameters reflecting higher GH/IGF-1 activity predisposed for radiographic progression. Especially patients with biochemical control by SMS analogs performed worse than surgically cured patients (adjusted OR=18.9,p=0.025). Clinical OA course varied strongly on individual level. Overall, self-reported hand function and grip strength deteriorated significantly, as well as self-reported lower limb function. Patients with high baseline pain or functional limitation scores showed a poor outcome of hand OA, poor outcome in lower limb OA was only predisposed by high BMI. Clinical change was not related to radiographic progression.[br][bold][italic]Conclusions[/italic][/bold][br]Acromegalic patients have progressive arthropathy, both clinically and radiographically, despite long-term biochemical control. Higher GH/IGF-1 activity was associated with radiographic progression. Remarkably, biochemical control by SMS analogs was associated with increased risk of radiographic progression, which may indicate insufficient GH control according to current criteria and the need of more aggressive therapy. Clinical and radiographic OA course were not related.[br][br]Nothing to Disclose: KC, SR, AP, JS, FR, HR, HK, MK, NB 2012-06-26T11:45:00 351 2012-06-26T00:00:00 1899-12-30T11:45:00 742 384 2773 OR41-3 OR22-01 Tuesday 2347 2012


2343 ENDO12L_OR41-4 ORAL SESSION: Novel Pituitary Tumor Syndromes [amp] Sellar Metastasis (11:15 AM-12:45 PM) High Prevalence of Vertebral Fractures in Acromegaly without Discernible Osteoporosis Jessica Brzana, Chris Yedinak, Nadia Hameed, Maria Fleseriu Oregon Health [amp] Science University, Portland, OR; Oregon Health [amp] Science University, Portland, OR; Oregon Health [amp] Science University, Portland, OR Introduction: The effects of excess GH/IGF-1 effects on bone mineral density (BMD), architecture and strength are not completely understood. Patients (pts) with acromegaly have mostly normal BMD and trabecular bone mass seems to depend more on gonadal status than GH as increased risk of vertebral fractures (VF) has been observed in both controlled and uncontrolled acromegaly. In the general population VF correlates with increased morbidity and mortality.[br]Aim: Assess the prevalence of VF in a US population of acromegaly patients.[br]Methods: A cross-sectional study in 34 consecutive acromegaly pts who underwent ant/post and lat, cervical, thoracic and lumbar spine X-rays evaluated using a Genant (Gen) method. Twenty-three pts had BMD data assessed by DEXA scan. Two-tailed t- test, one-way ANOVA, and bivariate correlations were performed using PAWS 18.[br]Results: Prevalence of VF was 38%, 13 pts exhibited a total of 35 VF (2.7/pt) (19 Gen 0.5, 9 Gen 1, 7 Gen 2). In the VF group (7M/6F), age at diagnosis was 43[plusmn]16 y, 69% were controlled by surgery, 1 by radiation and 5 by adjuvant medical therapy. No pt had DM, 2 pts were on hydrocortisone, and 2 were on testosterone. The study included 13 post-menopausal females, none on estrogen replacement. In the non-fracture (NF) group 4 women were on BCP. Based on BMD scores: no osteoporosis and osteopenia was found in 4 pts in each group (p=0.21) in the absence of any bone therapy. Disease duration in VF and NF was: 108[plusmn]75.2 and 82.2[plusmn]64.9 ms (ns). Estimated age of disease onset in the VF and NF group was 47[plusmn]16 and 45[plusmn]16 y (p=0.68), respectively. Acromegaly disease control [VF 69% vs NF 62% (p=0.30)] and tumor size were similar in both groups. Age, hypogonadism, urine NTX, alk phos, PTH, 25-OH vit D, T score were also no different between groups. However, vit D scores were inversely correlated with the Gen fracture score r=.550 (p=.004).[br]Discussion: Observed prevalence of VF (38%) was much higher than the 14% (p[lt]0.0005) reported for an age-matched general US population (1). Occurrence of VF risk was independent of BMD, hypogonadism, disease duration and control, DM, bone markers and vitamin D, although the latter may influence fracture severity.[br]Conclusions: Higher than expected VF prevalence in an unselected US population of acromegaly pts occurred despite absence of discernable osteoporosis. Accordingly, acromegaly pts should undergo spinal X-rays at diagnosis and follow-up, as dictated by clinical status.[br][br]Samelson EJ, Hannan MT, Zhang Y, Genant HK, Felson DT, Kiel DP. Incidence and risk factors for vertebral fracture in women and men: 25-year follow-up results from the population-based Framingham study. J Bone Miner Res. 2006 Aug;21(8):1207-14.[br][br]Nothing to Disclose: JB, CY, NH, MF 2012-06-26T12:00:00 351 2012-06-26T00:00:00 1899-12-30T12:00:00 594 384 2774 OR41-4 OR22-01 Tuesday 2348 2012


2344 ENDO12L_OR41-5 ORAL SESSION: Novel Pituitary Tumor Syndromes [amp] Sellar Metastasis (11:15 AM-12:45 PM) Diabetes Insipidus (DI), Headaches (HA) and Abnormal Eye Motility (AEM) in Patients with Sellar Masses Are Highly Predictive of Metastatic Disease to the Sella Ribal Al Aridi, Katia Elsibai, Pingfu Fu, Mehreen Khan, Dima Abdelmannan, Baha Arafah University Hospitals Case Medical Center and Louis Stokes VA Medical Center, Cleveland, OH; Case Western Reserve University, Cleveland, OH Metastatic tumors to sella are rare and often confused with other sellar masses such as pituitary adenomas. Hence, definitive diagnosis requires high degree of clinical suspicion. To date, no comprehensive studies have explored clinical predictors of metastatic disease to sella. The aim of this study is to characterize unique clinical features of this entity.[br][bold]Methods:[/bold] We reviewed all pathologically confirmed cases of pituitary/sellar metastasis published in the English and French literature and included our own experience for a total of 110 cases. Presenting features were compared with those of 38 surgically treated patients with non-secreting macroadenomas. ROC analyses were employed to evaluate DI, HA and AEM as diagnostic factors for metastasis. The area under ROC curve (AUC) was calculated to reflect the predictive value of each or combination of the 3 factors in identifying metastasis. Univariante and Multivariante analysis of predictors were performed.[br][bold]Results:[/bold] Similar gender and age (mean=56) distribution were noted among cases of sellar metastasis. In women, breast ca was the most common primary (15/55), followed by thyroid (9/55) and lung (9/55) cancers. Renal cell ca was the most common primary (10/55) in men followed by neuroendocrine (including Small Cell ca; 9/55), and other lung cancers in 7/55. Pituitary metastasis was the first manifestation of primary cancer in 25/55 women and 21/55 men. Presenting symptoms included HA (93%), visual field defects (VFD; 84%), AEM (84%) and DI (64%). Endocrine features included hypogonadism (69%), hypothyroidism (53%), adrenal insufficiency (47%), and hyperprolactinemia (61%). Patients with adenomas had similar age and gender distribution. Their presenting symptoms included HA (53%), VFD (39%), AEM (7%) but none had DI. Although the latter group had similar degree of hyperprolactenemia, it had lower rates of adrenal insufficiency (18%) and hypothyroidism (12%). HA, AEM and DI were significant (p[lt]0.0001) in predicting metastasis in univariate logistic regression analysis (odds ratio OR: 13, 63, 133 respectively). After adjusting for the effects of AEM in multivariate analysis, DI was still significant (OR: 50; p 0.026). Univariate ROC analysis showed an AUC of 0.82 for DI, 0.7 for HA and 0.88 for AEM. When these were considered simultaneously, AUC was 0.96 with sensitivity of 0.92 and specificity of 0.91.[br][bold]Conclusion:[/bold] The presence of DI, HA and AEM with a sellar mass accurately predicts metastatic disease.[br][br]Nothing to Disclose: RAA, KE, PF, MK, DA, BA 2012-06-26T12:15:00 351 2012-06-26T00:00:00 1899-12-30T12:15:00 1277 384 2775 OR41-5 OR22-01 Tuesday 2349 2012


2345 ENDO12L_OR41-6 ORAL SESSION: Novel Pituitary Tumor Syndromes [amp] Sellar Metastasis (11:15 AM-12:45 PM) Metastatic Neoplasms to the Sella: A Retrospective Chart Review Danit Ariel, Hotaik Tommy Sung, Nicole Coghlan, Robert Dodd, Iris Catrice Gibbs, Hannes Vogel, Laurence Katznelson Stanford University, Stanford, CA; Stanford University, Stanford, CA; Stanford University, Stanford, CA; Stanford University, Stanford, CA [italic]Background:[/italic] Metastatic disease to the sella is uncommon and there are limited available data regarding the demographics, clinical characteristics, and radiographic findings in such patients.[br][italic]Methods:[/italic] Retrospective chart review of adults at Stanford University Medical Center from 1980 to 2011 with metastatic disease to the sella.[br][italic]Results:[/italic] 14 subjects were identified (9 F). The mean age at diagnosis was 52 years (range: 19-73 y). 6 (43%) had breast carcinoma, 3 (21%) had renal cell carcinoma, 2 (14%) had squamous cell carcinoma of the head and neck, 1 had bronchoalveolar carcinoma of the lung, 1 had pineal germ cell tumor, and 1 had nodular sclerosing Hodgkin[apos]s lymphoma. The most common presenting symptom was headache (62%), followed by fatigue (58%), ophthalmoplegia (46%), and visual field defects (42%). 71% presented with at least one pituitary hormone insufficiency, including 7 (54%) with diabetes insipidus (DI). 6 (50%) subjects had secondary hypothyroidism, and 5 (45%) had secondary adrenal insufficiency. 8 (62%) tumors involved the stalk, 3 (23%) tumors involved the median eminence, and 6 (46%) invaded the cavernous sinus. 69% of the tumors demonstrated contrast enhancement on MRI. Of the patients with stalk involvement, 88% had DI. 93% of patients had a prior diagnosis of malignancy for a mean duration of 93 months (range: 8-276 mo). At diagnosis of sellar metastasis, 46% subjects had additional CNS lesions and 55% had extracranial metastases including lung, mediastinum, bone, liver, and lymph nodes. At the time of this review, there were three deaths, including two with breast cancer and one with renal cell carcinoma; time from diagnosis of the pituitary metastasis to death was 28 months (range: 17-37 mo).[br][italic]Discussion:[/italic] In this review of 14 patients with metastases to the sella, the most common neoplastic sources were breast and renal cell carcinoma. Systemic and/or other CNS disease was highly prevalent at the time of diagnosis. DI was the most common endocrine abnormality, followed by hypothyroidism, and adrenal insufficiency. Therefore, the presence of DI in patients with a known malignancy should alert concern for sellar metastases.[br][br]Disclosures: LK: Investigator, Novartis Pharmaceuticals, Corcept Therapeutics; Research Funding, Pfizer, Inc., Ipsen. Nothing to Disclose: DA, HTS, NC, RD, ICG, HV 2012-06-26T12:30:00 351 2012-06-26T00:00:00 1899-12-30T12:30:00 672 384 2776 OR41-6 OR22-01 Tuesday 2350 2012


2346 ENDO12L_OR42-1 ORAL SESSION: New Mediators of Obesity (2:00 PM-3:30 PM) [quot]Giant[quot] Protein AHNAK [mdash] Role in Obesity and Insulin Resistance Eddy Karnieli, Dafna Rozentzweig Ben-yosef, Maya Ramdas, Ahmad Assalia, Chava Harel, Irit Hochberg, Michal Armoni Rambam Medical Center, Haifa, Israel; Technion-Israel Institute of Technology, Haifa, Israel; Rambam Medical Center, Haifa, Israel Elevated levels of free fatty acids, like arachidonic acid (AA), are an important factor in the pathogenesis of insulin resistance and obesity. AA is known to suppress the GLUT4 promoter (G4P) specifically at -222/-197bp; however the mechanistic link is not yet understood. Applying Mass Spectrometric analysis to identify potential mediators of this mechanism, we have identified AHNAK, a giant phosphoprotein, in association with this specific G4P region in AA-treated cardiomyocytes. AHNAK is known to participate in hyperlipidemia-associated cardiac signaling with no known reference to obesity or insulin resistance.[br]To characterize the role of AHNAK in obesity/insulin resistance, we performed transient overexpression of AHNAK in primary rat adipocytes which repressed the G4P activity and the reverse effect was seen with silencing of AHNAK protein. Levels of AHNAK protein in aged/obese rats and ob/ob mice, models of insulin resistance/obesity, were significantly elevated (2 and 3-fold respectively) in adipose tissue, while Glut4 levels were concomitantly decreased (to 5 and 2-fold respectively), compared to lean controls. [br]Role of AHNAK in overall body metabolism was studied in AHNAK knockout mice (AHNAK-KO). AHNAK-KO at 6wks (fed normal chow) showed significant alterations in body composition compared to control mice, i.e., they had 50% less fat mass and higher lean body (muscle) mass (86 vs. 82% of body weight; p[lt]0.05). When challenged with high fat diet for 6 weeks, AHNAK-KO gained significantly less weight than controls (animal body weight: 29g vs. 38g; p[lt]0.05), thus demonstrating protection against diet-induced obesity.[br]Clinical contribution of AHNAK to obesity/insulin resistance was evaluated in adipose tissues obtained from obese and lean subjects undergoing bariatric surgery. In subcutaneous adipose tissues, the decrease in AHNAK mRNA levels was in significant correlation with the degree of body weight loss (R=0.943; p[le]0.005). In omental adipocytes, AHNAK protein levels negatively correlated with those of Glut4 (R=-0.810; p[lt]0.05). Of note, serum levels of AHNAK protein were significantly increased in obese and diabetic/obese patients by 1.9-fold and 2.4-fold respectively, compared to normal subjects and the levels also correlated with the BMI and HDL levels in these subjects.[br]In all, our data introduce AHNAK as a new contributor to the [ldquo]diabesity[rdquo] syndrome and therefore as a potential molecular target for obesity and type 2 diabetes therapy.[br][br]Nothing to Disclose: EK, DRB-Y, MR, AA, CH, IH, MA 2012-06-26T14:00:00 332 2012-06-26T00:00:00 1899-12-30T14:00:00 1407 398 2793 OR42-1 OR28-01 Tuesday 2351 2012


2347 ENDO12L_OR42-2 ORAL SESSION: New Mediators of Obesity (2:00 PM-3:30 PM) Female-Specific Obesity Caused by Conditional Ablation of Nkx2-1 in the Ventromedial Nucleus of the Hypothalamus Stephanie M Correa, David W Newstrom, Pierre Flandin-Blety, Clement C Cheung, John LR Rubenstein, Holly A Ingraham University of California San Francisco, San Francisco, CA; University of California San Francisco, San Francisco, CA; University of California San Francisco, San Francisco, CA The ventromedial hypothalamus (VMH) plays important roles in reproduction, ingestive behavior, anxiety, and aggression. [italic]Nkx2-1[/italic] is a homeobox transcription factor that is expressed in hypothalamic progenitors and persists in mature neurons of the VMH. To determine [italic]Nkx2-1[/italic] gene function in VMH neurons, we ablated [italic]Nkx2-1[/italic] specifically in the VMH using an [italic]Sf1Cre[/italic] driver. This strategy removes [italic]Nkx2-1[/italic] beginning at embryonic day 9. In adulthood, female [italic]Nkx2-1[sup]Sf1Cre[/sup][/italic] mutants were significantly heavier compared to control littermates when fed normal chow, whereas the body weight of male littermates was normal. Metabolic studies suggested that the higher body weight is due to reduced energy expenditure and not increased food intake. Specifically, locomotor activity is reduced in female [italic]Nkx2-1[sup]Sf1Cre[/sup][/italic] mice whereas basal metabolic rate and brown fat thermogenesis were normal. Global gene expression analyses of mutant female VMH confirmed that [italic]Nkx2-1[/italic] gene expression is significantly and dramatically reduced. Validation of the top up- and down-regulated hits by real time qPCR revealed a significant down-regulation of [italic]Esr1[/italic] and [italic]Nmu[/italic] in [italic]Nkx2-1[sup]Sf1Cre[/sup][/italic] VMH tissue samples. ER[alpha] (encoded by [italic]Esr1[/italic]) has been shown previously to regulate basal metabolic rate and brown fat thermogenesis in female mice. Central administration of the neuropeptide NMU has been shown to decrease food intake, increase activity levels and increase heat production in male and female rats. Spatial analysis of ER[alpha] and [italic]Nmu[/italic] expression confirmed a decrease in ER[alpha] and [italic]Nmu[/italic] in the ventrolateral VMH (VMH[sub]vl[/sub]) but normal expression elsewhere in the hypothalamus. Future studies will test the hypothesis that development of NKX2-1+, ER[alpha]+, and NMU+ neurons in the VMH[sub]vl[/sub] is essential for normal locomotor activity in female mice.[br][br]Nothing to Disclose: SMC, DWN, PF-B, CCC, JLRR, HAI 2012-06-26T14:15:00 332 2012-06-26T00:00:00 1899-12-30T14:15:00 2229 398 2794 OR42-2 OR28-01 Tuesday 2352 2012


2348 ENDO12L_OR42-3 ORAL SESSION: New Mediators of Obesity (2:00 PM-3:30 PM) Inhibition of the Non-Canonical IkB Kinases IKKe and TBK1 in Obese Mice Leads to Weight Loss and Amelioration of Metabolic Syndrome Irit Hochberg, Shian-Huey Chiang, Shannon Reilly, Dave Bridges, Alan Robert Saltiel University of Michigan, Ann Arbor, MI Obesity-induced activation of the inflammatory NFkB pathway has been identified as a major factor in the development of insulin resistance. The non-canonical IkB kinases TBK1 and IKKe, that are thought to be suppressors of the NFkB pathway, are induced by high fat diet after inflammation is initiated. We hypothesized that this counter-inflammatory induction is important during the development of obesity, and that blocking the effect of these kinases may lead to weight loss and amelioration of metabolic syndrome.[br]We demonstrate that pharmacologic inhibition of IKKe/TBK1 enhances inflammation in cultured adipocytes. Liver knockdown of IKKe and TBK1 expression by antisense oligonucleotides in high fat diet mice acutely increases liver and adipose tissue inflammation, while leading to weight loss and improved glucose tolerance. Administration of a novel small molecule IKKe/TBK1 inhibitor to obese mice also produced weight loss, improved glucose tolerance and attenuated hepatic steatosis. These effects were due to increased energy expenditure and not a reduction in food intake.[br]These results support the role of counter-regulatory pathways as an energy-preserving mechanism during the development of obesity. Disruption of this process may lead to an adaptive increase in energy expenditure and potentially be used to promote weight loss and reduce the co-morbidities of obesity.[br][br]Sources of Research Support: NIH R01DK060591.[br][br]Nothing to Disclose: IH, S-HC, SR, DB, ARS 2012-06-26T14:30:00 332 2012-06-26T00:00:00 1899-12-30T14:30:00 1161 398 2795 OR42-3 OR28-01 Tuesday 2353 2012


2349 ENDO12L_OR42-4 ORAL SESSION: New Mediators of Obesity (2:00 PM-3:30 PM) MMP12 Deficiency Is Associated with Increased Insulin Resistance and White Adipose Tissue Inflammation, but Not Increased Risk of Adiposity, in Mice Alli M Nuotio-Antar, C Wayne Smith Baylor College of Medicine, Houston, TX Obesity in humans and rodents results from an increase in white adipose tissue (WAT) depot size and is associated with increased risk for insulin resistance. WAT depot expansion is a dynamic process involving the differentiation and enlargement of adipocytes, angiogenesis, extracellular matrix (ECM) remodeling, and recruitment of immune cells, which can modulate the inflammatory and hypoxic environment of the expanding depot. Matrix Metalloproteinase 12 (MMP12) is a macrophage-secreted tissue elastase that may play a role not only in ECM remodeling, but also in inflammation, by selectively inactivating MCP1, which may recruit immune cells to the depot. In a preliminary study, MMP12 expression levels were elevated in Western diet-fed wildtype (wt) C57BL/6j mice in comparison to chow diet-fed wt controls, indicating that MMP12 is a gene that is selectively upregulated in WAT in the context of diet-induced obesity. In order to test the hypothesis that MMP12 impacts adiposity, WAT inflammation, and whole-body insulin sensitivity, we compared MMP12 knockout (MMP12-/-) mice with wt mice placed on WD or chow diet for 14 weeks. Initial studies using nonlittermate controls on WD indicated that MMP12-/- mice gained more weight, were more insulin resistant, and had increased expression of inflammatory genes in WAT than wt mice. Subsequent experiments conducted with littermate controls revealed no differences in body weight gain between MMP12-/- and wt mice placed on either chow diet or WD, although both MMP12-/- and wt mice gained more weight on WD diet than on chow diet. However, MMP12-/- mice were more insulin resistant than wt littermate controls, as determined by IPGTT, and had elevated fasting insulin levels on both chow and Western diets. TNF[alpha] expression and M1/M2 macrophage ratios, as determined by Nos2/Arg1 gene expression ratios, were increased in WD-fed MMP12-/- mice, relative to wt controls, suggesting that adipose tissue inflammation was greater in MMP12-/- mice than for wt littermate controls. Expression of adipocyte differentiation genes ap2 and PPAR[gamma] did not differ between MMP12-/- and wt controls fed the same diet, indicating that the process of adipocyte differentiation, per se, was not affected by MMP12 deficiency. Overall, our results suggest that MMP12 may play an important role in adipose tissue depot homeostasis, in particular by regulating adipose tissue inflammation, and thus impacting whole-body insulin resistance.[br][br]Nothing to Disclose: AMN-A, CWS 2012-06-26T14:45:00 332 2012-06-26T00:00:00 1899-12-30T14:45:00 1943 398 2796 OR42-4 OR28-01 Tuesday 2354 2012


2350 ENDO12L_OR42-5 ORAL SESSION: New Mediators of Obesity (2:00 PM-3:30 PM) Neuronal and Mechanistic Pathways Controlled by Neuropeptide Y (NPY) on the Thyrotropin-Releasing Hormone (TRH) Neuron Nicole E Cyr, Samantha Hyner, Mathew Sochat, Mario Perello, Ronald Stuart, Eduardo A Nillni The Warren Alpert Medical School of Brown University, Rhose Island Hospital, Providence, RI; Brown University, Providence, RI; Multidisciplinary Institute of Cell Biology, Buenos Aires, Argentina The hypothalamic pituitary thyroid (HPT) axis plays a critical role regulating metabolism and thermogenesis. Central regulation of the thyroid axis by thyrotropin releasing hormone (TRH) neurons in the hypothalamic paraventricular nucleus (PVN) is essential for normal function of the axis under different physiological conditions including cold stress and changes in nutritional status. Emerging evidence has highlighted the importance of leptin in regulating the HPT axis by increasing TRH production in the PVN. Leptin stimulates TRH directly in the PVN, and indirectly by enhancing alpha-melanocyte-stimulating hormone (alpha-MSH) production and inhibiting agouti-related peptide (AgRP) and neuropeptide-Y (NPY) in the hypothalamic arcuate nucleus (ARC). The current study reveals two mechanisms by which the potent appetite-stimulating NPY decreases TRH production wherein each mechanism opposes leptin[apos]s actions on TRH. Here we show for the first time that NPY decreases alpha-MSH peptide content in the ARC. We further demonstrate that NPY impairs the maturation of alpha-MSH from its precursor pro-opiomelanocortin (POMC). Specifically, NPY decreased the POMC processing enzyme prohormone convertase 2 (PC2), a process that is mediated by early growth response protein 1 (Egr-1). Moreover, this action of NPY on PC2 directly countered the Egr-1-mediated increase in PC2 induced by leptin. Earlier evidence revealed that NPY attenuates the alpha-MSH-induced increase in CREB signaling in TRH neurons indicating that NPY could oppose leptin[apos]s indirect actions on TRH. Here, we additionally demonstrated that NPY decreased CREB activation by both the potent alpha-MSH analogue melanotan II (MTII) and leptin. All together, results indicate a dual mechanism by which NPY reduces TRH (1) directly in the PVN and (2) indirectly via attenuated alpha-MSH production in the ARC. As each of these mechanisms counteracts leptin[apos]s actions on TRH production, results underscore the significance of the interaction between leptin and NPY in the central regulation of energy balance.[br][br]Sources of Research Support: NIDDK/NIH grant R01 DK085916-01 to EAN.[br][br]Nothing to Disclose: NEC, SH, MS, MP, RS, EAN 2012-06-26T15:00:00 332 2012-06-26T00:00:00 1899-12-30T15:00:00 964 398 2797 OR42-5 OR28-01 Tuesday 2355 2012


2351 ENDO12L_OR42-6 ORAL SESSION: New Mediators of Obesity (2:00 PM-3:30 PM) TR Agonists as Activators of Brown Fat Jean Z Lin, Alexandro J Martagon, Anisha A Gupte, Laurie J Minze, Willa A Hsueh, P Webb, J D Baxter, K J Phillips Methodist Hospital Research Institute, Houston, TX Although endogenous thyroid hormones, such as T3, are recognized as metabolic regulators, their use as anti-obesity agents is limited due to deleterious side effects. Here we show that synthetic thyroid hormone receptor agonists such as GC-1 are capable of eliciting dramatic weight loss in multiple mouse models of obesity. Severely obese (ob/ob) mice treated with GC-1 lose the majority of their body fat in as little as 10 days and experience a concomitant normalization of insulin sensitivity, cholesterol, and lipid levels due to a potent activation of brown adipose tissue (BAT) and induction of thermogenesis. Surprisingly, while obese mice treated with GC-1 exhibited striking thermogenesis, lean mice showed no elevation in metabolic rate and experienced no weight loss, suggesting that GC-1 may rescue a defect in adaptive thermogenesis unique to obese mice. Indeed, when challenged with either increased caloric intake or cold exposure, lean mice were able to induce brown fat activity in the absence of GC-1, while obese mice failed to do so unless treated with GC-1. In summary, select TR agonists including GC-1 appear to affect fat loss by rescuing obesity-related defects in brown fat activation. Given the renewed interest in the relationship between BAT activity and human obesity, TR agonists may warrant evaluation as anti-obesity therapeutics.[br][br]Sources of Research Support: This work was funded by NIH grant 1RC4DK090849.[br][br]Nothing to Disclose: JZL, AJM, AAG, LJM, WAH, PW, JDB, KJP 2012-06-26T15:15:00 332 2012-06-26T00:00:00 1899-12-30T15:15:00 2313 398 2798 OR42-6 OR28-01 Tuesday 2356 2012


2352 ENDO12L_OR43-1 ORAL SESSION: Novel Molecular Mechanisms of Diabetes [amp] Insulin Resistance (2:00 PM-3:30 PM) Fusion of Proinsulin-Producing Bone Marrow-Derived Cells with Neurons Underlies Diabetic Neuropathy Tomoya Terashima, Hideto Kojima, Hiroshi Urabe, Fan Lin, Lawrence Chan Baylor College of Medicine, Houston, TX; Shiga University of Medical Science, Tsukinowacho, Seta, Otsu, Japan; Shiga University of Medical Science, Tsukinowacho, Seta, Otsu, Japan Diabetic neuropathy is the most common diabetic complication. The dogma is that hyperglycemia-induced dysfunction and death of neurons, Schwann cells and vasa nervorum underlies diabetic neuropathy. Tumor necrosis factor (TNF)-[alpha] is a known pathogenic factor. We used streptozotocin (STZ) to induce diabetes in TNF-KO mice and found that, compared with wild-type (WT), TNF-KO mice are resistant to diabetic neuropathy. Further, treatment of WT diabetic mice with the anti-TNF monoclonal antibody infliximab prevents the neuropathy. We previously found that diabetes induces proinsulin-producing bone marrow-derived cells (PI-BMDCs) in multiple organs in rodents [Kojima et al PNAS 101: 2458, 2004]. Interestingly, these abnormal cells coexpress TNF-[Fujimiya et al PNAS 104: 4030, 2007]. We showed that PI-BMDCs fuse with neurons in the dorsal root ganglia (DRG) of diabetic mice, causing neuronal dysfunction and apoptosis [Terashima et al. PNAS 35: 12525, 2005]. Poly (ADP-ribose) polymerase (PARP) expression is another known pathogenic factor. We examined the relation between PARP, TNF-[alpha] and PI-producing BM-derived cells (PI-BMDCs) in the pathogenesis of diabetic neuropathy. We performed BM transplantation (BMT) of WT or PARPKO BM cells to WT or PARPKO mice and induced diabetes in the recipients using STZ. In diabetic WT mice, PARPKO BMT protects against diabetic neuropathy. As reported previously, PARPKO mice are resistant to diabetic neuropathy. Importantly, BMT of WT PARP+/+ BM and not PARPKO BM, conferred susceptibility to diabetic neuropathy to these normally resistant PARPKO mice. The severity of the nerve dysfunction in these models is correlated with the number of PI-BMDC-neuron fusion cells in the DRG.[br]A primary role of PI-BMDC- neuron fusion in diabetic neuropathy is supported by experiments [italic]in vitro[/italic]. Incubation of BM cells in high glucose induces PI and TNF-coexpression. Coincubation of isolated DRG neurons with PI-BMDCs in high glucose, but not low glucose, leads to fusion between the 2 cell types. Therefore, diabetic neuropathy is not caused merely by hyperglycemia-induced metabolic perturbations within neurons, as previously reported. Rather, a fundamental process in pathogenesis is the hyperglycemia-induced PARP- and TNF-[alpha]-coexpressing fusogenic PI-BMDCs and their subsequent fusion with DRG neurons. Abnormal BM cells are as much a culprit as dysfunctional neurons, Schwann cells and vasa nervorum in the pathogenesis of diabetic neuropathy.[br][br]Nothing to Disclose: TT, HK, HU, FL, LC 2012-06-26T14:00:00 342ABDE 2012-06-26T00:00:00 1899-12-30T14:00:00 955 399 2799 OR43-1 OR12-01 Tuesday 2357 2012


2353 ENDO12L_OR43-2 ORAL SESSION: Novel Molecular Mechanisms of Diabetes [amp] Insulin Resistance (2:00 PM-3:30 PM) Pro-Inflammatory State in Diabetes Mellitus: Critical Role of Increased Phosphodiesterase 4B (PDE4B) and Decreased Cellular cAMP Regulated LPS-Inducible TNF-[alpha] in Glucose-Primed Human Monocytes Leila Gobejishvili, Elaina M Chambers, Sri Prakash L Mokshagundam, Craig J McClain, Shirish S Barve University of Louisville, Louisville, KY; Robley Rex VA Medical Center, Louisville, KY Diabetes mellitus is a metabolic disorder that leads to the development of a number of complications. The etiology of each diabetic complication is undoubtedly multifactorial. The present work focuses on the critical component common in many diabetic complications i.e., dysregulation of innate immunity and increased inflammatory response. Both animal and clinical studies have shown that high glucose plays a significant pathogenic role in sustaining inflammation via increased expression of inflammatory cytokines such as TNF[alpha]. The underlying mechanisms have not been completely elucidated. The regulation of cytokines by cyclicAMP (cAMP) under hyperglycemic states has not been adequately addressed.[br]We used human monocytes from healthy individuals and cultured them under normal (5mM) and high (25mM) glucose concentrations, with or without lipopolysaccharide (LPS). Roliprim, a inhibitor of Phosphodiesterase 4B (PDE4B) was used to further eucidate the role of PDE4B. TNF[alpha] production and gene expression, PDE4B gene expression and activity and cellular cAMP levels were measured. Histone modifications at the TNF[alpha] and PDE4B promoter regions were analysed to determine potential role of epigenetic mechanisms in modulating cytokine responses.[br]Our data demonstrate that human PBMCs exposed to high glucose exhibit a [ldquo]primed phenotype[rdquo]; wherein they express hyper-elevated levels of TNF[alpha] following LPS stimulation. Notably, we show for the first time that HG exposure up-regulates PDE4B gene expression and PDE4B enzymatic activity leading to decreased cellular cAMP levels and increased TNF[alpha] expression. Moreover, selective inhibition of PDE4 significantly attenuated the HG mediated increase in TNF expression. We further determined that this priming phenomenon from HG exposure is the result of epigenetic changes involving histone modifications at the PDE4B and TNF[alpha] promoters leading to alterations in the relevant transcription factor binding (Cyclic AMP Response Element Binding protein and Nuclaear Factor [kappa]B).[br]These data strongly suggest that disruptions in cAMP/PDE4B homeostasis and signaling play a significant mechanistic/pathogenic role in the development of inflammation occurring in response to HG associated with diabetic complications. Moreover, these results indicate that PDE4B as a potential therapeutic target in the treatment of Diabetes.[br][br]Sources of Research Support: NIH-R01 Grant to Dr. Shirish Barve.[br][br]Nothing to Disclose: LG, EMC, SLM, CJM, SSB 2012-06-26T14:15:00 342ABDE 2012-06-26T00:00:00 1899-12-30T14:15:00 1797 399 2800 OR43-2 OR12-01 Tuesday 2358 2012


2354 ENDO12L_OR43-3 ORAL SESSION: Novel Molecular Mechanisms of Diabetes [amp] Insulin Resistance (2:00 PM-3:30 PM) Increased Pancreatic Auto-Inflammation in Young Rhesus Macaques on High-Fat Diet Lindsey E Nicol, Wilmon F Grant, Sarah M Comstock, M S Smith, Kevin L Grove, Daniel L Marks Oregon Health [amp] Science University, Portland, OR; Oregon Health [amp] Science University, Portland, OR; Oregon Health [amp] Science University, Portaland, OR Rationale: Deleterious effects of the obesity epidemic are found in pediatric population but we do not fully understand how to identify and qualify the early changes occurring in our young overweight patients. Obesity related inflammation is well recognized as a confounding factor of metabolic disease. Such auto-inflammation has been attributed to the pathophysiology of [beta]-cell dysfunction and the development of type 2 diabetes. Our interest is to help delineate how early auto-inflammation occurs in the spectrum of metabolic disease within the pancreas and the pancreatic islets. We hypothesize that cytokine expression and innate immune infiltration will be increased in the pancreatic tissue during the early phases of insulin resistance.[br]Objective: To identify and qualify the early changes of obesity related inflammatory markers in the pancreata of juvenile nonhuman primates (NHP) on chronic high fat diet (HFD).[br]Methods: Pancreata harvested from 13 month-old rhesus macaques exposed to chronic HFD from [italic]in utero[/italic] to necropsy were analyzed by quantitative PCR for cytokines identified with obesity-related inflammation and by immune histochemistry for islet-associated macrophages. Parameters from intravenous glucose tolerance testing, which reflected mild hyperinsulinemia, were correlated to the expression of the cytokines.[br]Results: Despite not yet having significant metabolic disease the HFD cohort had a two-fold increase in the relative expression of IL-6 ([italic]P[/italic] [lt] 0.036) and a sexually dimorphic increase in IL-1B ([italic]P[/italic] [lt]0.029) in males. Expression of IL-6 was strongly and positively associated with first phase insulin response (r[sup]2[/sup] = 0.83, [italic]P [/italic][lt] 0.012). Additionally, there were a greater number of islet-associated macrophages ([italic]P[/italic] [lt] 0.015) in the pancreata of the HFD cohort (8.3 [plusmn] 0.7) verses controls (5.1 [plusmn] 0.8 macrophages/islet area x 10[sup]-4[/sup] [micro]m[sup]2[/sup]) and a significant increase in the composite expression of all cytokines analyzed ([italic]P[/italic] [lt] 0.015).[br]Conclusion: Pancreatic tissue and islets from juvenile NHP exposed to HFD have increased inflammatory markers and evidence of innate immune infiltration prior to the onset of significant obesity or glucose dysregulation typically detected by standard clinic assays. Given the parallel development of obesity related metabolic disease between humans and NHPs these findings have strong relevance to the early obesity pathology driven by chronic HFD in children and adolescence.[br][br]Sources of Research Support: Child Health and Research Center Grant K12HD057588; 5R24DK090964-02.[br][br]Nothing to Disclose: LEN, WFG, SMC, MSS, KLG, DLM 2012-06-26T14:30:00 342ABDE 2012-06-26T00:00:00 1899-12-30T14:30:00 1869 399 2801 OR43-3 OR12-01 Tuesday 2359 2012


2355 ENDO12L_OR43-4 ORAL SESSION: Novel Molecular Mechanisms of Diabetes [amp] Insulin Resistance (2:00 PM-3:30 PM) Fatty Acid Amide Hydrolase Knockout Mice Manifest Decreased Whole-Body Energy Expenditure and Hepatic Insulin Resistance Whitney Harris Brown, Matthew Paul Gillum, Tiago Cardoso Alves, Derek Mark Erion, Hui-Young Lee, Jin Kwon Jeong, Ben Cravatt, Sabrina Diano, Gerald I Shulman Yale University, New Haven, CT; Yale University, New Haven, CT; The Scripps Institute, La Jolla, CA; Yale University, New Haven, CT A missense mutation of fatty acid amide hydrolase (FAAH) was found to be associated with obesity in humans, but the mechanism is unknown. In order to address this question we examined food intake, locomotor activity, whole body energy expenditure, and tissue-specific insulin sensitivity, assessed by the hyperinsulinemic-euglycemic clamp, in FAAH knockout mice and age-weight matched WT control mice fed either a regular chow or high-fat diet (HFD). On a regular chow diet, FAAH KO mice displayed a [sim]10% (P[lt]0.05) reduction in whole-body energy expenditure. When fed a HFD this reduction was associated with severe hepatic insulin resistance as reflected by reduced suppression of hepatic glucose production during the clamp (90% vs. 43%, P[lt]0.01). Hepatic insulin resistance in the FAAH KO mice could be attributed to an almost twofold increase in hepatic diacylglycerol content (372 nmol/g vs. 600 nmol/g, P[lt]0.01), which was associated with a 5 fold (P[lt]0.001) increase in PKC[epsilon] activity and a 60% (P[lt]0.01) reduction in insulin-stimulated IRS-2 tyrosine phosphorylation. Decreased energy expenditure in the FAAH KO mice could be attributed to decreased circulating T4 concentrations (3.0 ng/ml vs. 2.4 ng/ml, P[lt]0.0001) secondary to a 60% decrease in expression of thyroid stimulating hormone mRNA in the pituitary (P[lt]0.05), a 50% decrease in thyroid releasing hormone, and an 11-fold increase in deiodinase-2 mRNA expression in the hypothalamus (P[lt]0.05). All of these metabolic changes were replicated in mice chronically infused with two major n-acylethanolamine (NAE) substrates of FAAH; arachidonylethanolamine and palmitoylethanolamine. Taken together these data suggest that FAAH deletion predisposes mice to obesity and hepatic insulin resistance by increasing hypothalamic NAE concentrations leading to PPAR[gamma] activation in the hypothalamus which promotes hypothalamic deiodinase-2 expression resulting in central hypothyroidism.[br][br]Sipe et al.,International Journal of Obesity 2005; 755-759.[br][br]Sources of Research Support: NIDDK F31 DK083192-04; R01 DK-40936; U24 DK-059635; P30 DK-45735.[br][br]Nothing to Disclose: WHB, MPG, TCA, DME, H-YL, JKJ, BC, SD, GIS 2012-06-26T14:45:00 342ABDE 2012-06-26T00:00:00 1899-12-30T14:45:00 1894 399 2802 OR43-4 OR12-01 Tuesday 2360 2012


2356 ENDO12L_OR43-5 ORAL SESSION: Novel Molecular Mechanisms of Diabetes [amp] Insulin Resistance (2:00 PM-3:30 PM) Fractalkine, a Novel Cytokine, Is Associated with Adipose Tissue Insulin Resistance: Modulation of Plasma Fractalkine Levels by Low-Dose Pioglitazone Therapy in Type 2 Diabetes Zandra Perez-Cadena, Giuseppe Daniele, Alberto Chavez, Lauren Cortez, Amalia Gastaldelli, Ralph DeFronzo, Franco Folli, Devjit Tripathy University of Texas Health Science at San Antonio, San Antonio, TX; Audie L Murphy Veterans Hospital, San Antonio, TX Fractalkine (FRK), a novel chemokine, plays a role in inflammatory processes in the pathogenesis of atherosclerosis. Its role in human T2DM is however yet to be identified. Pioglitazone is an insulin sensitizer and has mild anti-inflammatory effects. We evaluated the relationship of plasma FRK levels and whole body and adipose tissue insulin resistance in T2DM and the effect of 6-month low-dose PIO (15 mg/day) therapy on plasma FRK in 20 type 2 diabetes (T2DM) patients (age: 56 [plusmn]1 yr, BMI: 33.5 [plusmn]1.3 kg/m[sup]2[/sup], FPG: 145 [plusmn]8, HbA[sub]1c[/sub]: 7.5 [plusmn]0.3%) randomized to PIO (n=11) or placebo (PLAC) (n=9). Subjects participated in an OGTT with glucose, insulin, C-peptide, and FFA measured every 30 minutes, euglycemic-hyperinsulinemic clamp (80mU/m[sup]2[/sup]) with measurement of FFA, adipose tissue IR (AT-IR=FFA [times] insulin) and body fat (BF) by DXA at 0 and 6 months. FRK was measured during OGTTs and insulin clamps and at 0, 1, 3, 5 and 6 months. PIO, but not PLAC, significantly improved FPG (-14% vs -5%), HbA[sub]1c[/sub] (-7% vs -3%), AT-IR (-47% vs +16%) and Insulin sensitivity (M value +0.9 vs +1.1 mg/kg/min) (all p[lt]0.02 vs PLAC). At baseline, fasting plasma FRK levels were 33[38] pg/ml (median [IQR range]), similar in PIO vs PLAC (33[30] vs 29[42] pg/ml, p=ns). At baseline, FRK was associated with AT-IR after correcting for BF (r=0.75, p=0.002). FRK levels did not change during OGTT. After 6 months, FRK decreased in subjects treated with PIO, both during the fasting state and during the insulin clamp (13[24] and 19[36] p[lt]0.01 vs baseline). On the other hand, FRK tended to increase during the clamp (40[34]; p=0.06 vs baseline, p[lt]0.003 vs PIO), with PLAC. Also, plasma FRK was higher throughout the 6-month period in PLAC vs PIO (51[69] pg/ml, mean of 6-months, p=0.007 vs baseline). The decrease in FRK after 6 months was associated with both a decrease in AT-IR after correcting for changes in BF and an improvement in glycemic control (r=0.62, p=0.05). In conclusion, plasma FRK is associated with AT-IR and PIO-mediated improvement in AT-IR could partly be due to decreased plasma FRK.[br][br]Sources of Research Support: Takeda Pharmaceutical Company; Audie L. Murphy Veterans Hospital.[br][br]Disclosures: RD: Principal Investigator, Amylin Pharmaceuticals, Takeda. FF: Research Funding, Takeda. Nothing to Disclose: ZP-C, GD, AC, LC, AG, DT 2012-06-26T15:00:00 342ABDE 2012-06-26T00:00:00 1899-12-30T15:00:00 1941 399 2803 OR43-5 OR12-01 Tuesday 2361 2012


2357 ENDO12L_OR43-6 ORAL SESSION: Novel Molecular Mechanisms of Diabetes [amp] Insulin Resistance (2:00 PM-3:30 PM) Insulin Resistance Is Associated with Inadequate Hepcidin Production Amir H Sam, Mark Busbridge, Anjali Amin, Lisa Webber, Davinia White, Stephen Franks, Niamh M Martin, Michelle L Sleeth, Nurhafzan A Ismail, Norlida Mat Daud, Dimitris Papamargaritis, Carel Le Roux, Richard S Chapman, Gary Frost, Stephen R Bloom, Kevin G Murphy Imperial College London, London, UK; Imperial College Healthcare NHS Trust, London, UK; Imperial College London, London, UK Background[br]The underlying mechanism for the increased body iron associated with insulin resistance is unclear. Hepcidin is the key hormone regulating iron homeostasis. We hypothesized that patients with insulin resistance have inadequate hepcidin production for their iron load.[br]Methods[br]Serum concentrations of the active form of hepcidin (hepcidin-25) and hepcidin: ferritin ratio were evaluated in two insulin resistant groups: patients with type 2 diabetes (n=33, controls matched for age, gender and BMI, n=33) and patients with polycystic ovary syndrome (PCOS, n=27, controls matched for age and BMI, n=16). To confirm that any changes observed were associated with insulin resistance rather than insulin deficiency, the same measurements were made in patients with type 1 diabetes (n=30, controls matched for age, gender and BMI, n=30). Finally, the relationship between homeostasis model assessment of insulin resistance (HOMA-IR) and serum hepcidin: ferritin ratio was explored in non-diabetic overweight/obese subjects (n=16).[br]Results[br]Patients with type 2 diabetes (p [lt]0.0005) and PCOS (p [lt]0.01) had a significantly lower hepcidin: ferritin ratio than controls. There was no significant difference in serum hepcidin: ferritin ratio between patients with type 1 diabetes and controls (p =0.43). Stepwise multiple regression analysis showed hepcidin: ferritin ratio to be an independent predictor of HOMA-IR in non-diabetic overweight/obese subjects ([szlig] =-0.42, p [lt]0.05).[br]Conclusions[br]Insulin resistance, but not insulin deficiency, is associated with inadequate hepcidin production. Reduced hepcidin synthesis may thus lead to increased body iron stores in insulin resistant states.[br][br]Nothing to Disclose: AHS, MB, AA, LW, DW, SF, NMM, MLS, NAI, NMD, DP, CLR, RSC, GF, SRB , KGM 2012-06-26T15:15:00 342ABDE 2012-06-26T00:00:00 1899-12-30T15:15:00 2024 399 2804 OR43-6 OR12-01 Tuesday 2362 2012


2358 ENDO12L_OR44-1 ORAL SESSION: Signaling in Tissue Development (2:00 PM-3:30 PM) KRAS Suppresses Endocrine Growth through a Menin-RASSF1 Regulatory Network Chester Chamberlain, Michael German University of California, San Francisco, CA KRAS is a regulated GDP/GTP molecular switch that controls several signaling networks. Although activating missense mutations in the KRAS gene are a major cause of accelerated growth in several epithelial cancers, KRAS signaling can elicit diverse, cell-type specific responses, and its function in normal tissue growth and physiology remains largely unknown. We discovered that mice carrying a heterozygous null mutation for KRAS (KRAS hets) and thus a decrease in KRAS signaling had accelerated growth of the pancreatic islet, parathyroid and pituitary glands, tissues known to develop tumors in patients with Multiple Endocrine Neoplasia type 1 (MEN1), a disorder caused by heterozygous null mutations in [italic]MEN1[/italic], the gene encoding the tumor suppressor Menin. Consistent with this observation, KRAS hets have improved glucose tolerance as adults due to increased numbers of insulin-producing beta-cells. The additional beta-cells come from two sources: an increased number of Neurogenin3-expressing endocrine progenitors during embryogenesis and accelerated beta-cell proliferation during the perinatal period. In contrast, increased KRAS signaling caused by expression of an activated KRAS allele specifically in the pancreas suppressed both sources of new beta-cells (neogenesis and proliferation) and activated in beta cells two opposing signaling arms downstream of KRAS: the pro-proliferative MAPK pathway and anti-proliferative RASSF1 pathway. We found that the dominance of the anti-proliferative KRAS effect in beta-cells depended on the expression of the endocrine tumor suppressor Menin. Our data suggest a model in which KRAS activates both the pro-proliferative MAPK pathway, and the anti-proliferative RASSF1 pathway. In the tissues susceptible to [italic]MEN1[/italic] gene mutation, Menin normally prevents the MAPK effector pathway from driving proliferation while leaving the inhibitory RASSF1 effector pathway intact. In this model, loss of Menin causes proliferation due to removal of the block in MAPK-driven proliferation downstream of KRAS, while loss of KRAS signaling increases proliferation by decreasing the unopposed RASSF1 activity in susceptible cells.[br][br]Sources of Research Support: This work is supported by:. The Larry L. Hillblom Foundation;. Nora Eccles Treadwell Foundation;. JDRF;. NIH/NIDDK.[br][br]Nothing to Disclose: CC, MG 2012-06-26T14:00:00 371 2012-06-26T00:00:00 1899-12-30T14:00:00 1390 400 2805 OR44-1 OR17-01 Tuesday 2363 2012


2359 ENDO12L_OR44-2 ORAL SESSION: Signaling in Tissue Development (2:00 PM-3:30 PM) Conditional Knockout of Activins in Adipose Tissues Has Profound Effects on Adiposity, Growth and Metabolism Chester Wayne Brown, Maria Namwanje, Juan Carlos Bournat, Lihua Huang Baylor College of Medicine, Houston, TX; Baylor College of Medicine, Houston, TX; Texas Children[apos]s Hospital, Houston, TX Body composition syndromes which include overweight, obesity as well as a variety of wasting syndromes associated with cancer, infectious and chronic diseases and their therapies, contribute substantially to human morbidity with a tremendous impact on the global economy. Our prior studies and those of others in humans and mouse models have implicated TGF-beta superfamily signaling as an important contributor the control of lean and fat body mass due to effects on energy expenditure, adipocyte differentiation and function. Using the Cre/Lox system, we have produced mice with conditional inactivation of activin beta A ([italic]Inhba[/italic]) in adipose tissues on an activin beta B ([italic]Inhbb[/italic]) null background. In contrast to perturbation of either activin alone, aP2-Cre, [italic]Inhba[/italic][sup]flox/-[/sup], [italic]Inhbb[/italic][sup]-/-[/sup] mice are small and have markedly reduced adiposity and survival, suggesting partially redundant metabolic effects originating from activins A and B in adipose tissues. Soluble ActRIIb-Fc receptors (used for their abilities to block activin receptor signaling) have shown promise in animal models to provide beneficial effects on body composition and energy expenditure. However, clinical trials in humans have revealed undesirable side effects. If activin receptor inhibition strategies are to be used therapeutically, it may prove helpful to understand the consequences of attenuating the biological functions of the individual ligands that signal through these receptors in the relevant cell types. In this way one can dissect the specific metabolic effects attributable to each, as well as side effects so as to develop the best strategy to elicit the most desirable metabolic outcomes.[br][br]Nothing to Disclose: CWB, MN, JCB, LH 2012-06-26T14:15:00 371 2012-06-26T00:00:00 1899-12-30T14:15:00 1290 400 2806 OR44-2 OR17-01 Tuesday 2364 2012


2360 ENDO12L_OR44-3 ORAL SESSION: Signaling in Tissue Development (2:00 PM-3:30 PM) Muscle Mass Maintenance in the Absence of Follistatin-Like 3 (FSTL3), an Inhibitor of Myostatin Andrew Lessey, Raymond Macharia, Douglas Donovan, Michelangelo Campanella, Abir Mukherjee Royal Veterinary College, London, UK Loss of muscle mass is associated with ageing and several degenerative pathological conditions which limit movement and critically impair quality of life. One key regulator of muscle mass is myostatin which inhibits muscle growth. In circulation, myostatin is bound to follistatin-like 3 (FSTL3) in an inactive complex. FSTL3 is a natural, soluble regulator of myostatin along with follistatin (FST) and myostatin propeptide. In the FSTL3 gene deleted mouse (FSTL3 KO), however, total body and muscle weights are not reduced. We therefore investigated whether myocellular regeneration is induced in the absence of FSTL3 to counter increased myostatin action, preventing muscle loss. Expression of FST is not induced in FSTL3 KO muscles, thus FST does not compensate for the absence of FSTL3. This suggests that active muscle generation might be increased in FSTL3 KO mice. Confirming this hypothesis, we found that while numbers of both satellite cells and activated satellite cells decline with age in both WT and FSTL3 KO mice, these populations are increased in FSTL3 KO mice compared to WT suggesting increased muscle generation in FSTL3 KO. Muscle twitch and tetanic force is not affected in FSTL3 KO mice demonstrating that functional muscles are maintained in FSTL3 KO mice. Active muscle generation increases glycolysis to retain energy balance and we characterized that ATP generation in myocytes from FSTL3 KO EDL muscles is indeed largely from glycolytic source. IGF is widely regarded as a key inducer of muscle growth in the adult and hence we examined whether IGF signalling is induced in FSTL3 KO muscle. While phosphorylation of protein kinase B (AKT), known to be activated by PI3 kinase downstream of the IGF receptor, is induced in FSTL3 KO muscles, IGF receptor activation is not. Importantly, we also show a reduction of PTEN, an inhibitor of PI3-kinase activity in FSTL3 KO muscle demonstrating a possible cross-talk between TGF[beta] ligand signalling and growth factor signalling via PTEN. This study demonstrates a novel mechanism involving increased AKT activation and increased satellite cell proliferation and activation that most likely mediates muscle mass maintenance in the absence of FSTL3, the physiological inhibitor of myostatin.[br][br]Nothing to Disclose: AL, RM, DD, MC, AM 2012-06-26T14:30:00 371 2012-06-26T00:00:00 1899-12-30T14:30:00 2083 400 2807 OR44-3 OR17-01 Tuesday 2365 2012


2361 ENDO12L_OR44-4 ORAL SESSION: Signaling in Tissue Development (2:00 PM-3:30 PM) Hepatocyte Growth Factor Promotes Osteogenic Differentiation of Human Mesenchymal Stem Cells through the p38 Pathway Kristina K Aenlle, Kevin M Curtis, Bernard A Roos, Guy A Howard Veterans Affairs Medical Center, Miami, FL; University of Miami MIller School of Medicine, Miami, FL; University of Miami MIller School of Medicine, Miami, FL; University of Miami MIller School of Medicine, Miami, FL Hepatocyte growth factor (HGF), also known as scatter factor, is a pleiotropic factor that regulates proliferation, motility, morphogenesis and angiogenesis. HGF has also been shown to participate in organogenesis, tissue repair, neuronal induction and bone remodeling [1]. HGF is secreted by human mesenchymal stem cells (hMSCs) and its autocrine/paracrine activity promotes the migration and proliferation of these cells. Our previous studies have shown that HGF treatment primes hMSC for osteogenic maturation [2]. As a follow-up to these seminal discoveries, we report here that HGF treatment of hMSC directly promotes osteogenic differentiation through the transcription of key osteogenic markers osteocalcin, osterix, bone salioprotein 2 (BSP2). Moreover, blocking the HGF signaling pathways using PHA 665752, an HGF receptor (c-Met) inhibitor, attenuates mineralization (as determined by Alizarin red-S staining) in hMSC. This data provides a direct link between HGF signaling and osteogenic maturation/differentiation.[br]HGF/c-Met promotion of osteogenic markers and support of mineralization is likely due to the rapid activation of the mitogen activated protein kinase (MAPK) signaling cascade. Of particular importance in this cascade is the p38 pathway. Originally described as a stress activating protein kinase, p38 is currently thought to have an important role in cell differentiation and apoptosis. p38 consists of four isoforms p38[alpha], p38[beta], p38[delta], and p38[gamma], and rapid induction of p38[alpha] and p38[beta] have been found to be important for skeletogenesis and bone homeostasis [3]. SB203580, a chemical inhibitor to p38[alpha] and p38[beta] showed a marked reduction in alkaline phosphatase (ALP) activity and expression, as well as a decrease in mineralization during hMSC osteogenic differentiation. Our novel results demonstrate that HGF promotes the rapid phosphorylation of total p38 in hMSC and a differential regulation of p38 isoform levels. RT-qPCR revealed that two days of HGF treatment increased the transcriptional level of p38[alpha], p38[beta] and p38[delta]; while decreasing the level of p38[gamma]. Western blot analysis showed that HGF treatment significantly increased the level of p38[beta] protein while having a moderate effect on the other p38 isoforms. While it is uncertain the role each isoform mediates during HGF induced hMSC osteogenic differentiation, our data demonstrates the importance of HGF and p38 signaling in osteogenic maturation/differentiation.[br][br]1. Grano, M., et al., Hepatocyte growth factor is a coupling factor for osteoclasts and osteoblasts in vitro. Proc Natl Acad Sci U S A, 1996. 93(15): p. 7644-8. 2. D[apos]Ippolito, G., et al., Cooperative actions of hepatocyte growth factor and 1,25-dihydroxyvitamin D3 in osteoblastic differentiation of human vertebral bone marrow stromal cells. Bone, 2002. 31(2): p. 269-75. 3. Greenblatt, M.B., et al., The p38 MAPK pathway is essential for skeletogenesis and bone homeostasis in mice. J Clin Invest. 120(7): p. 2457-73.[br][br]Sources of Research Support: VA Merit awarded to GAH and BAR.[br][br]Nothing to Disclose: KKA, KMC, BAR, GAH 2012-06-26T14:45:00 371 2012-06-26T00:00:00 1899-12-30T14:45:00 1939 400 2808 OR44-4 OR17-01 Tuesday 2366 2012


2362 ENDO12L_OR44-5 ORAL SESSION: Signaling in Tissue Development (2:00 PM-3:30 PM) Growth Hormone Receptor Gene Deletion in Mammary Epithelial Cells Leads to Impaired Pre-Pubertal Mammary Terminal End Bud Development but No Reduction in Primary PyVmT Epithelial Tumor Size Emily Jane Gallagher, Jeffrey Blank, Yingjie Wu, Lauren Rota, Teresa Wood, Derek LeRoith Mount Sinai School of Medicine, New York, NY; University of Medicine and Dentistry of New Jersey, Newark, NJ The growth hormone (GH)/insulin-like growth factor-1 (IGF-1) axis is important in mammary gland development and tumorigenesis. In humans, genetic defects in the GH receptor (GHR) are associated with decreased breast cancer prevalence; however, the direct effects of GH (distinct from those of IGF-1) on mammary epithelium and tumors in vivo has not been defined. Previously the effects of GH were thought to be mediated through its action on mammary stromal tissue, not directly on the epithelial cells (MEC). We aimed to study the role of GH in mammary gland development and tumorigenesis by deleting exon 4 of the GHR in the mammary epithelium using the MMTV-Cre (line A)/Lox system in FVB/N mice. We studied the role of GH on tumor growth and metastasis using the MMTV-Polyoma Virus middle T Antigen (PyVmT) transgenic model, hypothesizing that lack of GHR would reduce tumor growth and metastases. Mammary gland development was studied in mice heterozygous for MMTV-Cre and homozygous for the flox sites in the GHR (Cre+/- GHR LL), and 2 control groups: Cre heterozygous mice without the floxed GHR (Cre+/-) and mice with the floxed GHR, without Cre (Cre-/- GHRLL). Cre+/- GHRLL mice demonstrated cleavage of the floxed GHR in MEC, with some deletion in spleen and epidermis, similar to previous reports. Mice were characterized at age 3, 6 and 12 weeks. Whole mounts of mammary glands revealed fewer terminal end bud (TEBs) in 3 week Cre+/- GHR LL mice compared to the 2 control groups [Cre+/- GHR LL 0.9[plusmn]0.9; Cre+/- 12.4[plusmn]6.9; Cre-/- GHR LL 8.2[plusmn]2.1, p[lt]0.05]. Ductal invasion into the fat pad at 3 weeks was 3.7[plusmn]0.7mm in the Cre+/- GHRLL group, 4.4[plusmn]0.8mm in the Cre+/- and 4.1[plusmn]0.7mm in the Cre-/- GHR LL group (not significant). At 6 weeks no significant difference in number and size of TEBs or fat pad invasion was observed between the groups. No differences were observed in body weight or length between the groups. Tumor development was studied in PyVmT+/- Cre+/- GHR LL mice compared to PyVmT +/- Cre-/- GHR LL mice. No difference in primary tumor size or number of secondary tumors was seen between the 2 groups at 3 or 6 weeks. Our data demonstrate that GHR signaling may be important in epithelium for prepubertal TEB development, but after the onset of puberty other growth factors become prevalent. Primary tumor size is not affected by deleting the GHR in mammary epithelium. We are exploring how GH affects TEB cap cells and whether it alters the metastatic spread of tumors.[br][br]Nothing to Disclose: EJG, JB, YW, LR, TW, DL 2012-06-26T15:00:00 371 2012-06-26T00:00:00 1899-12-30T15:00:00 1256 400 2809 OR44-5 OR17-01 Tuesday 2367 2012


2363 ENDO12L_OR44-6 ORAL SESSION: Signaling in Tissue Development (2:00 PM-3:30 PM) The Metabolite GnRH-(1-5) Inhibits Migration in Immature GnRH Neurons Darwin Omar Larco, Madelaine Clark, T John Wu Uniformed Services University, Bethesda, MD; Uniformed Services University, Bethesda, MD The first five amino acids of the full-length GnRH are highly conserved across nonmammalian and mammalian species. In the extracellular matrix GnRH is metabolized by the zinc metallendopeptidase EC3.4.24.15 (EP24.15) to generate the pentapeptide GnRH-(1-5). Our previous studies have shown GnRH-(1-5) is biologically active in cultured GnRH neurons and facilitates lordosis behavior in female rats. However, the effect of GnRH-(1-5) if any on GnRH neuronal migration is not known. It is interesting to consider that GnRH-(1-5) may have a developmental function since EP24.15 is expressed in the CNS during development along the migratory path of GnRH neurons. To determine whether GnRH-(1-5) regulates cellular migration, we treated GN11 cells, a premigratory GnRH-secreting cell line, with GnRH-(1-5) and measured their migration using a wound-healing assay. Cells treated with 1 nM and 100 nM GnRH-(1-5) migrated significantly (p [lt] 0.05) less at 24h and 48h incubation periods relative to non-treated cells. Next, we sought to identify putative G protein-coupled receptors (GPCR) that bind GnRH-(1-5) using a high-throughput b-arrestin recruitment assay (DiscoverX, Fremont, CA). Interestingly, we found 4 candidate receptors, however, only the orphan receptor GPR173 was highly expressed in GN11 cells. To determine whether GPR173 is required for GnRH-(1-5) regulation of migration, GN11 cells were transfected with siRNA targeting GPR173 expression and the wound healing assay was repeated. Blocking GPR173 expression reversed the GnRH-(1-5) inhibition of migration. In contrast, cells transfected with a non-targeting siRNA pool had no effect on GnRH-(1-5) action, suggesting GnRH-(1-5) is likely acting through GPR173 to inhibit GN11 migration. To further characterize the downstream signaling of GnRH-(1-5), we looked at the STAT3 signaling pathway since this pathway has been implicated in the regulation of proliferation and migration. GN11 cells treated with 100 nM GnRH-(1-5) had a robust decrease (p [lt] 0.05) in phospho-STAT3 levels at 15 min (0.32[plusmn].22 relative to control cells normalized to 1) and levels were barely detectable at 60 min. In conclusion, GnRH-(1-5) may have a role in the development of the GnRH neuroendocrine system by regulating GnRH neuronal migration. Our results suggest this effect is mediated by the orphan receptor GPR173 and the STAT3 pathway.[br][br]Sources of Research Support: The National Science Foundation IOS-1052288; Department of Defense Grants RO85FN and TO85FU-01.[br][br]Nothing to Disclose: DOL, MC, TJW 2012-06-26T15:15:00 371 2012-06-26T00:00:00 1899-12-30T15:15:00 1932 400 2810 OR44-6 OR17-01 Tuesday 2368 2012


2364 ENDO12L_OR45-1 ORAL SESSION: Autoimmune Thyroid Disease (2:00 PM-3:30 PM) A Small Molecule Antagonist of TSH Receptor Inhibits Hyaluronic Acid Synthesis and Akt Phosphorylation Induced by Stimulatory Thyrotropin Receptor Antibodies in Graves Orbital Fibroblasts Adina F Turcu, Seema Kumar, Michael J Coenen, Susanne Neumann, Marvin C Gershengorn, Rebecca S Bahn Mayo Clinic, Rochester, MN; NIDDK, National Institutes of Health, Bathesda, MD [bold]Background:[/bold] Graves[apos] ophthalmopathy (GO) is an autoimmune disorder affecting the orbit. It is characterized by extraocular muscle and orbital adipose tissue expansion, as a consequence of hyaluronic acid (HA) accumulation with edema and new fat cell development. This process is thought to be triggered by autoantibodies targeting the TSH receptor (TSHR). We have shown that M22 and MS-1, two human monoclonal stimulatory TSHR antibodies (TSAb), increase HA synthesis and Akt phosphorylation in orbital fibroblasts from patients with GO. The current study was undertaken to determine whether a small molecule ligand antagonist of the TSHR (termed [quot][bold]1[/bold][quot]) might decrease the effect of M22 or MS-1 treatment on HA synthesis and Akt phosphorylation in these cells.[br][bold]Methods:[/bold] Orbital fibroblasts from patients with GO were grown to confluence and serum starved for 24 hr. Cells intended for inhibition experiments were pretreated with[bold] 1[/bold] for 24 h or 30 min. Cultures were then treated with M22 (100 ng/ml; n=7), MS-1 (10 ug/ml; n=6) or untreated in control media, with or without[bold] 1[/bold] (30 uM). HA was measured at 48 h, while pAkt, was determined at 10, 30 and 60 min, using commercially available ELISA kits.[br][bold]Results:[/bold] Treatment of GO orbital fibroblast cultures with M22 or MS-1 resulted in significantly increased secretion of HA (mean 1.34 fold, SEM: 0.13, p[lt]0.001, and mean 1.28 fold, SEM: 0.04, p=0.002, respectively) and pAkt (mean 4.09 fold, SEM: 0.8, p=0.002 and mean 3.29 fold, SEM: 1.0, p=0.002, respectively, at 30 minutes). Addition of[bold] 1[/bold] resulted in significant inhibition of M22 or MS-1 stimulated HA synthesis (69% reduction, SEM: 10.4%, p=0.001 and 36.9% reduction, SEM: 9.6%, p=0.004, respectively) and Akt phosphorylation (50%, SEM: 12.9%, p=0.022 and 45%. SEM: 9%, p=0.032, respectively, at 30 minutes).[br][bold]Conclusion:[/bold] A small molecule antagonist of TSHR inhibits TSAb-stimulated HA synthesis and Akt phosphorylation in GO orbital fibroblasts. Similar TSHR ligands may in future prove effective as therapy for GO.[br][br]Nothing to Disclose: AFT, SK, MJC, SN, MCG, RSB 2012-06-26T14:00:00 370 2012-06-26T00:00:00 1899-12-30T14:00:00 1638 401 2811 OR45-1 OR40-01 Tuesday 2369 2012


2365 ENDO12L_OR45-2 ORAL SESSION: Autoimmune Thyroid Disease (2:00 PM-3:30 PM) Cytokines and Peroxisome Proliferator-Activated Receptor [alpha] Agonists Modulate [beta] (CCL2) and [alpha] (CXCL10) Chemokines in Graves Ophthalmopathy Alessandro Antonelli, Silvia Martina Ferrari, Stefano Sellari-Franceschini, Alda Corrado, Caterina Mancusi, Moira Ferrini, Ele Ferrannini, Poupak Fallahi University of Pisa, School of Medicine, Pisa, Italy; University of Pisa, Pisa, Italy Introduction. No study has evaluated the effect of cytokines on the prototype beta chemokine (C-C motif) ligand (CCL)2 in Graves[apos] ophthalmopathy (GO), nor of peroxisome proliferator-activated receptor (PPAR)alpha activation on this chemokine secretion in fibroblasts or preadipocytes in GO.[br]Methods. We tested the interferon (IFN)gamma and tumor necrosis factor (TNF)alpha effect on CCL2, and for comparison on the prototype alpha chemokine (C-X-C motif) ligand (CXCL)10, and the possible modulatory role of PPARalpha activation on these chemokines secretion in normal and GO fibroblasts or preadipocytes in primary cell cultures.[br]Results. We show that IFNgamma alone, or in combination with TNFalpha was able to stimulate the secretion of CCL2 in primary orbital fibroblasts or preadipocytes from patients with GO, at levels similar to those observed in controls. IFNgamma and TNFalpha stimulated also CXCL10 chemokine secretion as expected. The presence of PPARalpha and [ndash]gamma in primary fibroblasts or preadipocytes from patients with GO has been confirmed. PPARalpha activators were able to inhibit the secretion of CXCL10 and CCL2, while PPARgamma activators were confirmed to be able to inhibit CXCL10, but had no effect on CCL2. PPARalpha activators were stronger inhibitors of chemokines secretion than PPARgamma agonists.[br]Conclusions. CCL2, and CXCL10, are modulated by IFNgamma and TNFalpha in GO. PPARalpha activators are able to inhibit the secretion of the main prototype alpha (CXCL10) and beta (CCL2) chemokines in GO fibroblasts or preadipocytes, suggesting that PPARalpha may be involved in the modulation of the immune response in GO.[br][br]Nothing to Disclose: AA, SMF, SS-F, AC, CM, MF, EF, PF 2012-06-26T14:15:00 370 2012-06-26T00:00:00 1899-12-30T14:15:00 1499 401 2812 OR45-2 OR40-01 Tuesday 2370 2012


2366 ENDO12L_OR45-3 ORAL SESSION: Autoimmune Thyroid Disease (2:00 PM-3:30 PM) A Bioassay for Thyroid-Stimulating Autoantibodies Is Clinically Useful and Predictive for Response to Therapy in Patients with Graves Disease George J Kahaly, Sabine Niel, Sarah Stannek, Kristina Bischof, Julia Kochanowski, Tanja Diana, Michael Kanitz, Paul D Olivo Gutenberg University Medical Center, Mainz, Germany; Diagnostic Hybrids, Inc, Athens, OH [bold]Background and aims: [/bold]Scarce data exist on the role of thyroid stimulating autoantibodies (TSAb) in the management of Graves[apos] disease (GD). Using an [italic]in vitro[/italic] diagnostic bioassay for measurement of TSAb, we hypothesized that TSAb levels are clinically useful and predictive in GD.[br][bold]Methods:[/bold] A total of 91 untreated hyperthyroid patients with GD (73 female) were treated with antithyroid drugs (ATD) for 24 weeks. Serum TSAb levels were measured at baseline, at 4, 8, 12, 24 and 36 weeks after starting ATD with a chimeric TSHR Bioassay (Thyretain) and expressed as the percent specimen-to-reference-ratio (SRR%). Response vs. non-response to treatment were defined as biochemical euthyroidism at weeks 24 (end of therapy) and 36 vs. persistent hyperthyroidism at week 24 and/or relapse at week 36.[br][bold]Results: [/bold]In this ongoing prospective clinical trial, 54 hyperthyroid patients with GD (40 female, median age 43 years, range 19-74 years, 31 with orbitopathy, GO) have completed to date the 24 week ATD treatment (Methimazole 2.5-20 mg/day, titration regimen) and the subsequent 12 week follow up. 23/54 (43%) responded to ATD treatment of whom 10/23 (43.5%) had GO. In contrast, 31/54 (57%) were non-responders, 21/31 (68%) with GO. As early as 12 weeks after start of ATD therapy, highly significant differences of ATD dose and serum TSAb levels were noted between responders vs. non-responders (p[lt]0.001). At week 24, the median ATD dose was 3-fold higher in responders vs. non-responders (2.5 vs. 7.5 mg/day, p[lt]0.001). Serum TSAb levels decreased markedly from week 0 to week 24 in responders (median SRR% 374 vs. 261, minus 30%, p=0.002), in contrast TSAb did not change in non-responders (SRR% 416 vs. 446, delta 7.0%, p=0.103). At week 24, median TSAb was SRR% 467 vs. 261 in non-responders vs. responders (p[lt]0.001). Median thyroid volume was 17.3 ml (range 5.4-35.7) vs. 22.7 ml (5-64) in responders vs. non-responders. Five of six male responders had a thyroid volume [lt]20 ml whereas it was [gt]40ml in 5/6 male non-resp. (p=0.008). Serum levels of TSAb and TSHR binding inhibiting immunoglobulins (Elecsys TRAb, Roche) closely correlated (week 24, r=0.62, p[lt]0.001). Positive correlations were also seen at week 24 between serum TSAb levels and serum fT4 (r=0.36, p=0.007) and fT3 (r=0.62, p[lt]0.001) values.[br][bold]Conclusions: [/bold]Response to ATD is accompanied by a significant decrease in TSAb levels. In contrast, TSAb levels remain unchanged or increase in non-responders, most of whom have systemic GD.[br][br]Disclosures: GJK: Research Funding, Diagnostic Hybrids, Inc. PDO: Employee, Diagnostic Hybrids, Inc. Nothing to Disclose: SN, SS, KB, JK, TD, MK 2012-06-26T14:30:00 370 2012-06-26T00:00:00 1899-12-30T14:30:00 1444 401 2813 OR45-3 OR40-01 Tuesday 2371 2012


2367 ENDO12L_OR45-4 ORAL SESSION: Autoimmune Thyroid Disease (2:00 PM-3:30 PM) Anatabine, a Tobacco Alkaloid, Reduces Disease Incidence and Severity in a Mouse Model of Autoimmune Thyroiditis Alessandra De Remigis, Melissa Anne Landek-Salgado, Marcella Ferlito, Shintaro Iwama, Schey-Cheng Tzou, Paul Ladenson, Patrizio Caturegli The Johns Hopkins Medical Institutions, Baltimore, MD; The Johns Hopkins Medical Institutions, Baltimore, MD; The Johns Hopkins Medical Institutions, Baltimore, MD Introduction: Tobacco smoking has been shown in epidemiological studies to favorably influence the course of some autoimmune diseases like ulcerative colitis and Hashimoto thyroiditis. This effect perhaps occurs through nicotine, which is known to possess immunosoppressive proprieties. Nicotine, however cannot be used due to its toxicity, addictive properties, and short half-life. We used anatabine, a tobacco alkaloid of tobacco that has a more favourable pharmacological profile, in a mouse model of experimental autoimmune thyroiditis (EAT).[br]Materials and methods: Experimental autoimmune thyroiditis (EAT) was induced in 61 CBA/J female mice by injection of mouse thyroglobulin emulsified in Freund[apos]s adjuvant. Anatabine, provided by Rock Creek Pharmaceuticals-Washington DC, was added to the drinking water of 29 mice at a concentration of 0.05mg/ml, yielding a daily dose of 12.5 mg/Kg of body weight. Control mices (No.= 32) drank regular water. Mice were bled on days 0, 7, 10, 14, and 21 after immunization and sacrificed on day 21 to assess thyroid histopathology, serum thyroxine (T4), and thyroglobulin antibodies. Anatabine effects were also assessed in vitro using a macrophage cell line (RAW 264.7) stimulated with interferon gamma (10 nM).[br]Results: Anatabine administration significantly reduced the incidence and severity of EAT. 31 of 32 (97%) control mice developed EAT, as compared to 21 of 29 (72%) anatabine treated mice (p= 0.007 by chi squared).The mean graded histopathological severity was 1 in the anatabine group and 1.5 in the water control group (p= 0.028 by Wilcoxon ranksum test). Thyroglobulin antibodies developed in both groups of mice starting on day 10 post-immunization, furhter increasing on day 14 and day 21. Anatabine treated group had significantly lower antibody levels on day 14 post-immunization, but similar levels on day 10 and 21. Serum T4 levels were significantly ameliorated by anatabine administration on day 21 post-immunization (5.25 vs 3.65 mcg/dL, p= 0.034); no differences were found in the preceeding time points. Anatabine suppresssed in a dose-dependent manner the induction of pro-inflammatory markers like inducible nitric oxide synthase and cyclooxygenase 2 in response to interferon gamma.[br]Conclusions: We have shown that anatabine ameliorates the thyroid lesions and hypothyroidism typical of experimental autoimmune thyroiditis.Further studies are needed to elucidate its mechanisms of action and potential clinical utility.[br][br]Nothing to Disclose: ADR, MAL-S, MF, SI, S-CT, PL, PC 2012-06-26T14:45:00 370 2012-06-26T00:00:00 1899-12-30T14:45:00 2174 401 2814 OR45-4 OR40-01 Tuesday 2372 2012


2368 ENDO12L_OR45-5 ORAL SESSION: Autoimmune Thyroid Disease (2:00 PM-3:30 PM) Activating Autoantibodies to the Beta 1 Adrenergic and M2 Muscarinic Receptors Facilitate Atrial Fibrillation in Patients with Graves Hyperthyroidism Allison Lea Galloway, Megan Danielle Vanderlinde-Wood, Muneer Ahmad Khan, Li Hongliang, Caitlin Marie Zillner, Yu Xichun, David Charles Kem University of Oklahoma and Veterans Administration Medical Centers, Oklahoma City, OK [bold]Objective: [/bold]To expand a study which previously revealed that activating autoantibodies to [beta]1 adrenergic receptor ([beta]1AR) and to muscarinic M2 receptor (M2R) may facilitate atrial fibrillation (AF) in Graves[apos] hyperthyroidism (1).[br][bold]Methods: [/bold]76 patients including 31 with Graves[apos] hyperthyroidism and AF, 36 with Graves[apos] and no known AF, and 9 with toxic multinodular goiter (TMNG) including one with AF. Peptides from the second extracellular loop of human [beta]1AR and M2R were synthesized and used to coat ELISA plates. O.D. at 60 min from 1:100 dilutions of sera were compared to that from 10 normal control subjects.[br][bold]Results: [/bold]45% of patients with Graves[apos] hyperthyroidism and AF were were positive for autoantibodies to [beta]1AR and 65% of the patients were positive for autoantibodies to M2R. Twelve subjects harbored both autoantibodies. The youngest patient to demonstrate AF was 38 yrs in thyroid storm. All others were [gt]45 yrs. Autoantibodies were present in 42% of the hyperthyroid subjects without AF. Patients with TMNG had a low prevalence of autoantibodies to these autonomic receptors. The single patient with TMNG and AF had normal autoantibody levels directed toward [beta]1AR and M2R.[br][bold]Discussion: [/bold]Activating autoantibodies to [beta]1 and M2 autonomic receptors are elevated in many patients with Graves[apos] hyperthyroidism and concurrent AF and to a lesser extent in those without known AF (1). [beta]1AR and M2R agonists and thyroid hormone facilitate atrial tachyarrhythmias. Hyperthyroidism has been associated with tachyarrhythmias and sustained AF in 20-30% of patients even after treatment. The prevalence of AF in patients with Graves[apos] disease increases with age. These autoantibodies to [beta]1AR and M2R have been demonstrated previously to increase intracellular Ca2+, shorten the action potential and hyperpolarize atrial cells. This combined sympathetic and parasympathetic stimulation generates early afterdepolarizations and rapid triggered firing in the pulmonary veins which in turn induces AF. Patients with TMNG in contrast to those with Graves[apos] hyperthyroidism have a relatively low prevalence of these activating autoantibodies compatible with the non-autoimmune pathogenesis of their hyperthyroidism.[br][bold]Conclusion: [/bold]This study supports and expands our previous clinical evidence that activating autoantibodies to [beta]1AR and M2R are associated with atrial tachyarrhythmias and may facilitate atrial fibrillation in older subjects with Graves[apos] hyperthyroidism.[br][br]Stavrakis et al. JACC 2009;54; 1309.[br][br]Nothing to Disclose: ALG, MDV-W, MAK, LH, CMZ, YX, DCK 2012-06-26T15:00:00 370 2012-06-26T00:00:00 1899-12-30T15:00:00 2305 401 2815 OR45-5 OR40-01 Tuesday 2373 2012


2369 ENDO12L_OR45-6 ORAL SESSION: Autoimmune Thyroid Disease (2:00 PM-3:30 PM) Significance of Maintaining Euthyroid State with Ultra-Low-Dose Methimazole (MMI) for More Than 3 Years To Attain Remission of Graves Disease Hirotsugu Kawamoto, Ayumi Kurihara, Naoki Edo, Hisanori Dambara, Koji Morita, Yuji Tanaka National Defense Medical College, Tokorozawa, Japan; National Defense Medical College, Tokorozawa, Japan Background: MMI is most widely used for treatment of Graves[apos] disease (GD) in Japan, because its advantage over other modalities is significant even with several disadvantages such as adverse effects (WBC, liver, skin, etc), longer treatment and high recurrence rate after discontinuation of the drug.[br]Purpose: To make a better criterion for discontinuation of MMI especially that for patients with GD near remission.[br]Patients and Methods: Patients (F:20 M:3, aged 56+-14) who kept euthyroid and negative TRAb (measured as TBII) with MMI 10 mg/w or less for more than 2 years were asked whether or not to continue MMI. Both [ldquo]continuation[rdquo] group #1 (14 cases aged 62+-11) and [ldquo]discontinuation[rdquo] group #2 (9 cases aged 49+-14) were followed up for longer than another 2 years. Elevation of FT4 or suppression of TSH after exclusion of [ldquo]destructive thyroiditis[rdquo] was judged as [ldquo]relapse[rdquo]. There was no significant difference in clinical profiles between 2 groups, such as TRAB or dose of MMI which varied from 10mg/month to 10mg/week.[br]Results: First, only 1 out of 14 cases in #1 relapsed who needed adjustment of dose of MMI, whereas 5 out of 9 cases in #2 were judged [ldquo]relapse[rdquo] within 21 months and were restarted with MMI. Although most patients who relapsed in #2 showed marked or considerable increase in TRAb, fluctuation of TRAb in #1 did not lead to increase in FT4 except one relapse mentioned above.[br]Discussion: Our observations provide several important clinical points: 1. patients under ultra low dose MMI who seem to be in remission still have considerable disease activity; 2. MMI at such small dose still has crucial effects to suppress GD activity. We believe its mechanism is mostly due to inhibition of hormone synthesis that shut down potential vicious cycle of autoimmune activity. Regarding the optimal duration of treatment, we believe it to be longer than three years, because autoimmune activity of GD is influenced by a variety of yearly or seasonal factors such as age, hay fever, pregnancy, mental instability and smoking, and because two years was not sufficient for safe discontinuation in this study.[br]Conclusion: Taken together that adverse effect of MMI with this small dose should be negligible and that discontinuation of MMI could cause more frequent hospital visit and blood examination, we would like to propose that patients with GD must be maintained in euthyroid state with ultra low dose ([lt] 10mg/w) of MMI for more than 3 years before discontinuation of treatment.[br][br]Nothing to Disclose: HK, AK, NE, HD, KM, YT 2012-06-26T15:15:00 370 2012-06-26T00:00:00 1899-12-30T15:15:00 1464 401 2816 OR45-6 OR40-01 Tuesday 2374 2012


2370 ENDO12L_OR46-1 ORAL SESSION: Biomarkers of Tumorigenesis [amp] Tumor Progression (2:00 PM-3:30 PM) Plasma Kisspeptin Is a Novel Biomarker of Tumor Metastasis in Patients with Ovarian Carcinoma Alexander N Comninos, Channa N Jayasena, Adam Januszewski, Hani Gabra, Alexandra Taylor, Chioma Otti, Richard Harvey, Mohammad A Ghatei, Stephen R Bloom, Waljit S Dhillo Imperial College London, London, UK; Imperial College Healthcare NHS Trust, London, UK; Imperial College Healthcare NHS Trust, London, UK Over 15,000 women in the USA die from ovarian carcinoma annually (1). The key determinant of patient survival is tumor stage at diagnosis. However currently available biomarkers such as CA125 are poor at differentiating between stages (2). It is therefore of critical importance to develop novel biomarkers of ovarian carcinoma stage which can help to stratify patients according to prognosis and guide their treatment.[br]Kisspeptin, encoded by the KISS1 gene, is a peptide activating the kisspeptin receptor[italic]. KISS1[/italic] tissue expression negatively correlates with metastasis risk in many tumors (3). In ovarian carcinoma, low [italic]KISS1[/italic] expression is associated with aggressive disease and increased expression of tissue invasion factors (4,5). Conversely [italic]KISS1[/italic] over-expression inhibits cell migration and reduces metastasis in ovarian carcinoma (4,5).[br]We compared plasma kisspeptin and serum CA125 levels in healthy controls, patients with stage 1 (no metastasis) and patients with stage 2-4 (metastasis) ovarian carcinoma (62 samples total), in order to determine the utility of plasma kisspeptin as a novel biomarker of tumor metastasis in ovarian carcinoma.[br]Mean plasma kisspeptin was significantly elevated in patients with stage 1 ovarian carcinoma when compared with patients with stage 2-4 ovarian carcinoma and controls (pmol/l: stage 1 25.1[plusmn]4.8, stage 2-4 11.8[plusmn]2.1, [italic]P[/italic][lt]0.05 vs. stage 1; healthy controls 13.1[plusmn]1.2, [italic]P[/italic][lt]0.01 vs. stage 1). In patients with ovarian carcinoma, a plasma kisspeptin cut-off [gt]20pmol/l was 91.3% specific and 44.4% sensitive for stage 1 disease. However a plasma kisspeptin cut-off [lt]10pmol/l was 100% specific and had improved sensitivity at 61.2% with respect to the more advanced stage 2-4 ovarian carcinoma. By comparison, a serum CA125 cut-off [gt]1000U/ml was 100% specific but only 26.1% sensitive when identifying the more advanced stage 2-4 ovarian carcinoma ([italic]vs.[/italic] stage 1). Receiver-operator characteristic (ROC) analysis of plasma kisspeptin in stage 2-4 vs. stage 1 ovarian carcinoma had an area under curve 0.80.[br]This study suggests that patients with stage 1 ovarian carcinoma have significantly elevated levels of plasma kisspeptin when compared with patients with stage 2-4 ovarian carcinoma or healthy controls. Furthermore plasma kisspeptin may be more reliable in distinguishing between stage 1 vs. stage 2-4 disease in comparison to CA125. Plasma kisspeptin testing may therefore provide a simple, rapid and non-invasive novel method to aid ovarian cancer staging.[br][br](1) Howlader N, Noone AM, Krapcho M, et al. SEER Cancer Statistics Review, 1975-2008, National Cancer Institute. Bethesda, MD, http://seer.cancer.gov/csr/1975_2008/, based on November 2010 SEER data submission, posted to the SEER web site, 2011. (2) Bast RC, Badgwell D, Lu Z, et al. New tumor markers: CA125 and beyond. Int J Gynecol Cancer. 2005; 15:274-81. (3) Beck HB, Welch DR. The KiSS-1 metastasis suppressor: A good night kiss for disseminated cancer cells. European Journal of Cancer. 2010; 46:1283-9. (4) Hata K, Dhar DK, Watanabe Y, et al. Expression of metastin and a G-protein-coupled receptor (AXOR12) in epithelial ovarian cancer. European Journal of Cancer. 2007; 43:1452-1459. (5) Gao GL, Liu LD, Zou XS, Chen WX. Expression of KiSS-1, matrix metalloproteinase-9, nuclear factor-kappaBp65 in ovarian tumours. Zhonghua Fu Chan Ke Za Zhi. 2007; 42:34-8.[br][br]Sources of Research Support: The Section is funded by the MRC, BBSRC, NIHR, an Integrative Mammalian Biology (IMB) Capacity Building Award, FP7-HEALTH-2009-241592 EuroCHIP grant and the NIHR Imperial Biomedical Research Centre. Other funding support: NIHR Clinical Lectureship(C.N.J); AMS/Wellcome Starter Grant(C.N.J); SFE Early Career Grant(C.N.J); ICHNT Charity Fellowship(A.N.C); Wellcome/GSK Fellowship(A.N.C); NIHR Career Development Fellowship(W.S.D).[br][br]Nothing to Disclose: ANC, CNJ, AJ, HG, AT, CO, RH, MAG, SRB, WSD 2012-06-26T14:00:00 360 2012-06-26T00:00:00 1899-12-30T14:00:00 173 402 2817 OR46-1 OR43-01 Tuesday 2375 2012


2371 ENDO12L_OR46-2 ORAL SESSION: Biomarkers of Tumorigenesis [amp] Tumor Progression (2:00 PM-3:30 PM) Genetic Labeling and Lineage Tracing of the Epithelial Stem Cells during Mouse Prostate Development, Regeneration, Tumorigenesis and Castration-Resistant Tumor Growth Li Gao, Jean Ching-Yi Tien, Suoling Zhou, Lan Liao, Jianming Xu Baylor College of Medicine, Houston, TX Recent studies showed that p63-expressing prostate basal stem cells (BSCs) are multipotent epithelial progenitors that have tumor-initiating potential when transplanted in mouse renal graft. However, the roles of basal stem cells during development, maintenance and regeneration of prostate epithelium have not been studied under physiological conditions, and whether the androgen-insensitive BSCs could be origin of the castration-resistant prostate cancer (CRPC) remains unknown.[br]For genetic labeling and gene manipulation in BSCs, we generated a p63CreERT2 knock-in mouse model that expresses tamoxifen-inducible Cre in the p63-expressing basal epithelial cells of the prostate. Lineage-tracing in the prostate of P63CreERT2;Rosa26-lacZ bigenic mice revealed that BSCs give rise to AR-positive luminal epithelial cells (LECs) and AR-negative neuroendocrine cells (NECs) during prostate growth. Moreover, lineage-labeled BSCs in the regressed prostate of castrated mice gave rise to LECs after androgen-induced prostate regeneration. Interestingly, a population of BSC-derived intermediate cells (IMCs)/LECs exhibited castration resistance and remains in the epithelium for a long time after castration, suggesting that BSCs and their immediate offspring might render castration resistance in prostate tumor.[br]We then generated P63CreERT2;Ptenf/f;Rosa26-lacZ tumor mouse model. Orthotopic injection of 4-hydroxytamoxifen into the mouse prostate induced Pten deletion in BSCs, leading to remarkable growth of prostate tumor and metastasis. These lineage-labeled tumor cells derived from BSCs strongly expressed androgen receptor (AR), with the highest cell proliferation levels in basal and intermediate epithelial cell layers, suggesting that the Pten-null BSCs and intermediate cells (IMCs) are tumor-initiating cells in this mouse model. Moreover, castrated tumor mice exhibited temporary shrink of the prostate tumor, while sustained significant proliferation in the basal/intermediate layers. Despite of androgen depletion, these tumor-initiating BSCs/IMCs expressed high levels of AR and AR target genes, recapitulating the molecule features of human CRPC. These results prove that BSCs generate all types of differentiated epithelial cells and maintain epithelium homeostasis in the mouse prostate, and that Pten mutation in prostate BSCs leads to castration-resistant tumor growth, highlighting the role of BSC population as a key target for new therapy against advanced prostate cancer.[br][br]Sources of Research Support: NIH R01 Grant DK058242.[br][br]Nothing to Disclose: LG, JC-YT, SZ, LL, JX 2012-06-26T14:15:00 360 2012-06-26T00:00:00 1899-12-30T14:15:00 2381 402 2818 OR46-2 OR43-01 Tuesday 2376 2012


2372 ENDO12L_OR46-3 ORAL SESSION: Biomarkers of Tumorigenesis [amp] Tumor Progression (2:00 PM-3:30 PM) Metformin Inhibits Growth and Decreases Resistance to Anoikis in Medullary Thyroid Cancer Cells Joanna Klubo-Gwiezdzinska, Kirk Jensen, John Costello, Jr, Aneeta Patel, Andrew Bauer, Kenneth Dale Burman, Leonard Wartofsky, Vasyl V Vasko Washington Hospital Center/Georgetown University, Washington, DC; Uniformed Services University of the Health Sciences, Bethesda, MD [bold]Introduction. [/bold]Medullary thyroid cancer (MTC) is associated with activation of ERK and mTOR signaling pathways. Recent studies showed that both ERK and mTOR can be inhibited by activated 5[apos]AMP-activated protein kinase (AMPK). The anti-diabetic agent metformin decrease proliferation of various cancer cells through AMPK dependent down-regulation of mTOR signaling.[br][bold]Objectives. [/bold]In the current study we assessed the [italic]in vitro [/italic]effect of metformin on proliferation and activation of mTOR, ERK, AKT and AMPK signaling pathways in MTC derived cells. We also examined the expression of molecular targets of metformin in human MTC.[br][bold]Methods.[/bold] We performed growth, viability, migration and resistance to anoikis assays using two MTC-derived cell lines (TT and MZ-CRC-1). Expressions of molecular targets of metformin were examined in MTC cell lines by Western blot and in 14 human MTCs tissue samples by immunostaining.[br][bold]Results. [/bold]Metformin inhibited growth and decreased expression of Cyclin D1 in MTC cells. Treatment with metformin was associated with inhibition of mTOR/p70S6K/pS6 signaling and down -regulation of pERK in both TT and MZ-CRC-1 cells. Metformin had no significant effects on pAKT in examined cell lines. Metformin inducible AMPK activation was noted only in TT cells. Treatment with AMPK inhibitor (Compound C) or AMPK silencing did not prevent growth inhibitory effects of metformin in TT cells. Metformin had no effect on MTC cell migration, but reduced their ability to form multi-cellular spheroids in non adherent conditions.[br]Immunostaining of human MTC showed over-expression of Cyclin D1 in all tumors compared to corresponding normal tissue. Activation of mTOR/p70S6K was detected in 8/14 (57.1%) of examined tumors.[br][bold]Conclusions[/bold]: Together these findings demonstrate that growth inhibitory effects of metformin in MTC cells are associated with down-regulation of both mTOR/p70S6K and pERK signaling pathways. Expression of the molecular targets of metformin in human MTC suggests its potential utility for the treatment of MTC patients.[br][br]Nothing to Disclose: JK-G, KJ, JC, AP, AB, KDB, LW, VVV 2012-06-26T14:30:00 360 2012-06-26T00:00:00 1899-12-30T14:30:00 1100 402 2819 OR46-3 OR43-01 Tuesday 2377 2012


2373 ENDO12L_OR46-4 ORAL SESSION: Biomarkers of Tumorigenesis [amp] Tumor Progression (2:00 PM-3:30 PM) The Inheritance of Polymorphisms of Genes Related to the Metabolism and Detoxification of Estrogen Influences the Susceptibility to Differentiated Thyroid Carcinoma Natassia Elena Bufalo, Aline Carolina Nadai Silva, Mariana Bonjiorno Martins, Raquel Bueno Barbieri, Fernando Assis Batista, Ligia Vera Montalli Assumpcao, Laura Sterian Ward University of Campinas, Campinas, Brazil The inheritance of genetic polymorphisms related to the metabolism and detoxification of estrogen plays an important role in a series of diseases[apos] susceptibility and could help explain differences in prevalence and behavior of differentiated thyroid carcinomas (DTC) among genders. We employed TaqMan SNP Genotyping to study [italic]CYP17A1, HSD17[beta]-1, CYP1A1, CYP1A2, CYP1B1, COMT [/italic]and[italic] SULT1E1[/italic] polymorphisms in 292 DTC patients (248 women and 44 men, 42.23[plusmn]14.81 years old) compared to 308 healthy controls (246 women and 62 men, 36.86[plusmn]12.95 years old). DTC was associated to menopaused women (p[lt]0.0001) and hormone replacement therapy (p[lt]0.0001). The inheritance of polymorphic homozygous [italic]CYP17A1[/italic] gene increased the susceptibility to DTC (OR=1.648, 95%CI=1.018-2.669; p=0.0421) while the heterozygous variant was associated with higher number of abortions (p=0.0150). The inheritance of CT genotype of [italic]CYP1A1m1 [/italic]and AC genotype of [italic]CYP1A2*F [/italic]increased the susceptibility to DTC (OR=1.302,95%CI=0.906-1.872,p=0.0328; OR=1,612,95%CI=1.129-2.300;p=0.0085, respectively) and is also associated with earlier age of DTC diagnoses (p=0.0073). The inheritance of a 119 TT [italic]CYP1B1[/italic] increased the risk for DTC compared with the GG genotype (OR=4.009,95%CI=2.468-6.513;p[lt]0.0001) and with the GT genotype (OR=2.153,95%CI=1.448-3.201;p=0.0002), beeing associated with smoking (p=0.0269), menopause (p=0.0317), hormone replacement therapy (p=0.0197) and menarche age (p=0.0077). A multivariate regression analysis showed that codon 432 CG [italic]CYP1B1 [/italic]also increased the risk to DTC (OR=1.697, 95%CI=1.165-2.472; p=0.0059) and was associated with the risk for papillary carcinomas (p=0.0243). There was no relationship between codon 453 [italic]CYP1B1[/italic] polymorphisms and the susceptibility to DTC, but we found an association between contraceptive use and the CC [italic]CYP1B1[/italic] genotype (p=0.0332). We did not observe any relationship between [italic]HSD17[beta]-1[/italic], [italic]COMT[/italic] and [italic]SULT[/italic] genes and the susceptibility to DTC neither to their clinical features. We demonstrated that exogenous and endogenous features related to sex hormones have considerable individual variability. Different individual polymorphic inheritances, which are attributed to polymorphisms of genes encoding enzymes involved in the production, metabolism and elimination of estrogen, may define subpopulations of women who are affected by increased exposure to estrogens and their metabolites, which can affect cell growth and induce cellular damage, hence increasing the susceptibility to DTC.[br][br]Sources of Research Support: FAPESP, CAPES.[br][br]Nothing to Disclose: NEB, ACNS, MBM, RBB, FAB, LVMA, LSW 2012-06-26T14:45:00 360 2012-06-26T00:00:00 1899-12-30T14:45:00 1847 402 2820 OR46-4 OR43-01 Tuesday 2378 2012


2374 ENDO12L_OR46-5 ORAL SESSION: Biomarkers of Tumorigenesis [amp] Tumor Progression (2:00 PM-3:30 PM) Igf-I Inhibition by Pasireotide Reverses the Mammary Phenotype of a Novel Mouse Model of Brca1 Deficiency Pietro Ameri, Ignacio Fernandez-Garcia, Haydeliz Martinez-Ruiz, Cristina De Angelis, Mary Helen Barcellos-Hoff, David L Kleinberg New York University School of Medicine, New York, NY; New York University School of Medicine, New York, NY; University of Genova, Genova, Italy BRCA1 germline mutations confer a substantial risk of breast cancer, attributed to a trifecta of defective DNA repair, genomic instability, and abnormal mammary lineage commitment.[br]We developed a novel mouse model of Brca1 deficiency, characterized by an extremely abnormal mammary phenotype. At 28 days of age, affected animals had more numerous and larger terminal end buds, in which cell proliferation rate was significantly higher compared to wildtype mice (p[lt]0.01). As animals aged, their mammary glands exhibited ductal dilatation, cystic changes, pre-malignant hyperplastic lesions, and ductal carcinoma in situ. An index of genomic instability based on in situ analysis of centrosomes was significantly increased in Brca1-deficient epithelium compared to wildtype tissue (p[lt]0.01). We identified a population of mammary basal epithelial cells expressing both luminal and myoepithelial lineage markers that represented 22.5% of the basal cell population in Brca1-deficient mice, compared to 3.9% in wildtypes (p[lt]0.01).[br]Phosphorylation of IGF-I receptor (IGF-IR) and its downstream signaling mediators were increased in affected mice. To determine if IGF-I inhibition would reduce or reverse mammary gland abnormalities, we tested the effects of pasireotide, a somatostatin analog that inhibits IGF-I action by a direct effect in the mammary gland and also by reducing serum GH. In 4-month-old mice, 7 days of treatment with pasireotide (vs. vehicle) caused a reduction in primary, secondary, and tertiary duct width (for primary ducts: 109.2 [mu]m with pasireotide vs. 313.8 [mu]m with vehicle, p[lt]0.01), halved the epithelial cell proliferation rate (10.6% vs. 20.8%, p[lt]0.05), normalized the centrosome aberration index (1.1 vs. 2.3, p=0.01), and eliminated the aberrant epithelial population (4.8% vs. 22.5%, p[lt]0.01). Phosphorylation of IGF-IR and its downstream mediators was inhibited. Results were similar after treatment for 7 days with PQ401, a small molecule inhibitor of IGF-IR, substantiating the IGF-I dependence of the Brca1-deficient phenotype response to pasireotide.[br]In conclusion, we present a novel mouse model of Brca1 deficiency with pre-malignant and early malignant changes, which is extremely sensitive to short-term IGF-I inhibition by pasireotide. Translation of these findings may prove effective in preventing breast cancer in women with BRCA1 mutations.[br][br]Disclosures: DLK: Medical Advisory Board Member, Novartis Pharmaceuticals; Researcher, Novartis Pharmaceuticals; Principal Investigator, Novartis Pharmaceuticals. Nothing to Disclose: PA, IF-G, HM-R, CDA, MHB-H 2012-06-26T15:00:00 360 2012-06-26T00:00:00 1899-12-30T15:00:00 824 402 2821 OR46-5 OR43-01 Tuesday 2379 2012


2375 ENDO12L_OR46-6 ORAL SESSION: Biomarkers of Tumorigenesis [amp] Tumor Progression (2:00 PM-3:30 PM) BRCA1 Regulates GR Signaling Pathway in Human Breast Cells Myriam Vilasco-Gamberoni, Laudine Communal, Justine Hugon-Rodin, Aurelie Courtin, Najat Mourra, Sylvie Dumont, Frederique Pennault-Llorca, Patricia Forgez, Anne Gompel INSERM, UPMC, Paris, France; Universit[eacute] Paris Descartes, APHP Hotel Dieu, Paris, France; APHP, Saint Antoine, Paris, France; Centre Jean Perrin, Clermont-Ferrand, France Inherited germline mutations in the [italic]BRCA1[/italic] tumour suppressor gene confer a genetic predisposition to breast and ovarian cancers. Most of the studies have explored [italic]BRCA1[/italic] interaction with estradiol and progesterone dependent signalling pathway, demonstrating that [italic]BRCA1[/italic] can regulate the transcriptional activity and degradation of estradiol/progesterone receptors. Among the hormones involved in breast cancer progression, less importance has been devoted so far to Glucocorticoids (GCs). GCs can increase the proliferation of normal breast cells but are antiproliferative on breast cancer cells (1). Inflammation can either trigger or decrease breast cancer progression. Based on the importance of GCs signalling in the regulation of breast cell homeostasis, inflammation and the implication of stress in breast cancer progression, we investigated the potential interplay that can occur in [italic]BRCA1[/italic] mutated patients. In this study, we demonstrated a loss of expression of the Glucocorticoid Receptor (GR) active form, GR P-Ser211, [italic]in[/italic] [italic]vivo[/italic] in normal breast tissues from BRCA1 mutation carriers compared to normal breast tissues from non-mutated patients. Furthermore, we observed a drastic loss of GR P-Ser211 expression in a series of triple negative breast cancers with BRCA1 proven mutation compared with sporadic triple negative tumors. We showed that BRCA1 is essential to develop an efficient GC-dependent signalling pathway, as its overexpression favoured MAPK p38 phosphorylation and further activation resulting in an amplified phosphorylation of p38 known target GR Ser211. BRCA1-mutant patients might develop an attenuated GR signalling pathway which can be associated with a higher proliferative rate and derepression of inflammatory processes. GR P-Ser211 phosphorylation staining might constitute an important diagnostic factor for screening loss of expression [italic]BRCA1[/italic] whether by germline mutation or epigenetic modifications in BRCAness tumours.[br][br](1) Vilasco M et al, Breast Cancer Res Treat. 2011;130:1-10.[br][br]Sources of Research Support: This research was supported by grants from INSERM-UPMC, HRA Pharma and the Institut National du Cancer (INCa, France). Laudine Communal was the recipient of a CIFRE grant from HRA Pharma and the government. Myriam Vilasco was the recipient of a post-doc fellowship (INCa). Aur[eacute]lie Courtin was a recipient of a grant from the Association pour la Recherche sur le Cancer.[br][br]Nothing to Disclose: MV-G, LC, JH-R, AC, NM, SD, FP-L, PF, AG 2012-06-26T15:15:00 360 2012-06-26T00:00:00 1899-12-30T15:15:00 106 402 2822 OR46-6 OR43-01 Tuesday 2380 2012


2376 ENDO12L_OR47-1 ORAL SESSION: Causes [amp] Consequences of Androgen Excess in Women (2:00 PM-3:30 PM) The Role of Adiponectin and Its Receptors in Theca Cell Androgen Production: Relevance to PCOS Fabio V Comim, James Turton, Kate Hardy, Stephen Franks Institute of Reproductive and Developmental Biology (IRDB), London, UK; Federal University of Santa Maria (UFSM), Santa Maria, Brazil [bold]Background: [/bold]Polycystic ovary syndrome (PCOS) is the commonest endocrine disorder in women, being characterized by ovarian hyperandrogenism. Androgen synthesis is enhanced in the presence of obesity, thus raising the question of a possible role for adipokines as an endocrine link between metabolic and reproductive abnormalities in PCOS. Clinical studies have shown that serum levels of adipokines may indeed be perturbed in PCOS but few studies have addressed the possibility of direct effects of adipokines on reproductive tissues[sup]1,2[/sup].[br][bold]Aims: [/bold]The aims of this study were: 1) to explore the effects of adipokines on ovarian steroidogenesis by bovine theca cells in culture; and 2) to analyse the expression of adiponectin receptors in archived ovarian tissue from PCOS and normal women.[br][bold]Methods: [/bold]1) Primary culture of bovine theca cells: immunoassay for androstenedione; expression of genes in androgen steroidogenic pathway; effects of knockdown of adiponectin receptors on androgen production; 2) Sections of archived human ovaries from normal women (n=10) and PCOS women (n=15): immunohistochemistry for adiponectin receptors 1 (ADIPOR1) and 2 (ADIPOR2)[br][bold]Results: [/bold]In bovine theca cells treated with LH (10 ng/ml), the administration of adiponectin (3 [italic][micro][/italic]g/ml), but not other adipokines (resistin 300 ng/ml, visfatin 600 ng/ml or leptin 600 ng/ml), resulted in decreased androstenedione production ([italic]P[/italic][lt] 0.0001 ANOVA). Adiponectin treatment was accompanied by a significant reduction of gene expression of key steroidogenic elements ([italic]CYP11A1, CYP17A1, STAR[/italic] and [italic]LHR[/italic]) ([italic]P[/italic] [lt]0.02). Moreover, the knockdown of genes for ADIPOR1, ADIPOR2 and its common mediator APPL1, was associated with higher rates of androstenedione secretion by bovine theca cells ([italic]P[/italic]=0.04 Kruskal-Wallis).[br]In human ovarian tissue, the proportion of theca cells expressing ADIPOR1 and ADIPOR2 was significantly lower in polycystic compared with normal ovaries ([italic]P [/italic]= 0.018 and [italic]P[/italic]=0.04 respectively[italic],[/italic] Mann-Whitney).[br][bold]Conclusions: [/bold]In summary, these results provide evidence of a link between fat cell metabolism and ovarian steroidogenesis, suggesting a role of disruption of adiponectin and/or its receptors in the pathogenesis of hyperandrogenism in PCOS.[br][br](1) Lagaly DV et al., Mol Cell Endocrinol 2008; 284:38-45. (2) Spicer L et al., Anim Reprod Sci 2011; 124:19-27.[br][br]Sources of Research Support: MRC; Capes Foundation.[br][br]Nothing to Disclose: FVC, JT, KH, SF 2012-06-26T14:00:00 372 2012-06-26T00:00:00 1899-12-30T14:00:00 1090 403 2823 OR47-1 OR34-01 Tuesday 2381 2012


2377 ENDO12L_OR47-2 ORAL SESSION: Causes [amp] Consequences of Androgen Excess in Women (2:00 PM-3:30 PM) Blunted Day-Night Changes of Luteinizing Hormone Pulse Frequency in Postmenarcheal Girls with Obesity: Potential Role of Hyperandrogenemia Jessicah SP Collins, Michelle Y Abshire, Jennifer P Beller, Christine M Burt Solorzano, R Jeffrey Chang, John C Marshall, Christopher R McCartney University of Virginia Health System, Charlottesville, VA; University of California, San Diego, La Jolla, CA Day-night changes of luteinizing hormone (LH) (and by inference GnRH) pulse frequency are well described during puberty. Early puberty is marked by low daytime LH frequency with an overnight increase, while late puberty is characterized by higher daytime frequency with an overnight decrease (1, 2). Day-night changes of LH pulse frequency are blunted in obese (OB) early pubertal girls (3, 4). We hypothesized that altered day-night LH patterns are also observed in late pubertal OB girls, and that such alterations may be influenced by hyperandrogenemia (HA), a common finding in OB girls.[br]We studied 63 post-menarcheal girls: ages 9-17 y (mean [plusmn] SD, 15.0 [plusmn] 1.7 y), Tanner stages 3-5 (4.7 [plusmn] 0.5), BMI-for-age-percentile 16.5[ndash]99.7 (83.6 [plusmn] 21.0). LH pulse frequency was determined by 10 min blood sampling over 12 h (1900[ndash]0700 h). Subjects were awake 1900[ndash]2300 h ([ldquo]day[rdquo]), but typically asleep 2300[ndash]0700 h ([ldquo]night[rdquo]). The primary outcome was day-to-night change of LH pulse frequency, and girls with OB (BMI-for-age percentile [ge] 95) were compared to girls without OB. We also assessed OB girls alone, comparing those with HA (by Tanner stage-specific norms) to those without HA. Data are reported as percentage (mean [plusmn] SEM) day-to-night change of LH frequency.[br]Non-OB (n=36) and OB (n=27) girls had similar daytime LH frequencies, and both demonstrated significant overnight decreases in LH pulse frequency; however, non-OB girls had a more pronounced day-to-night decrease of LH frequency compared to OB girls (percent change [ndash]30 [plusmn] 3 vs. [ndash]11 [plusmn] 5, p=0.016).[br]OB girls with and without HA also had similar daytime LH frequencies. OB girls without HA exhibited a significant day-to-night decrease of LH frequency (percent change [ndash]19 [plusmn] 12, p=0.039, n=9), but OB girls with HA did not (percent change [ndash]8 [plusmn] 6, p=0.645, n=18). Although OB non-HA girls appeared to have more overnight slowing in LH frequency than OB HA girls, this was not demonstrable statistically.[br]In conclusion, day-night changes of LH pulse frequency are blunted not only in early pubertal OB girls, but also in late pubertal OB girls. These data suggest that this phenomenon is exacerbated by HA.[br][br](1) Boyar R et al., N Engl J Med 287:582-586, 1972. (2) Apter D et al., J Clin Endocrinol Metab 76:940-949, 1993. (3) McCartney C et al., J Clin Endocrinol Metab 94: 56-66, 2009. (4) Bordini B et al., J Clin Endocrinol Metab 94:1168-1175, 2009.[br][br]Sources of Research Support: NIH F32 HD066855[ndash]01 awarded to JSPC; NIH U54 HD28934 awarded to JCM; NIH NICHD R01 HD058671-01 awarded to CRM.[br][br]Nothing to Disclose: JSPC, MYA, JPB, CMBS, RJC, JCM, CRM 2012-06-26T14:15:00 372 2012-06-26T00:00:00 1899-12-30T14:15:00 2222 403 2824 OR47-2 OR34-01 Tuesday 2382 2012


2378 ENDO12L_OR47-3 ORAL SESSION: Causes [amp] Consequences of Androgen Excess in Women (2:00 PM-3:30 PM) [beta]-Cell Dysfunction and Hyperandrogenemia in PCOS Early Pubertal Female Relatives Laura C Torchen, Naomi R Fogel, Wendy J Brickman, Nelly Mauras, Andrea Dunaif Children[apos]s Memorial Hospital, Chicago, IL; Feinberg School of Medicine, Northwestern University, Chicago, IL; Mayo College of Medicine, Jacksonville, FL First-degree female relatives (FDRs) of women with polycystic ovary syndrome (PCOS) are at substantially increased risk for PCOS. Although PCOS cannot be diagnosed until adolescence, there is emerging evidence that the process may begin earlier. We investigated the development of reproductive and metabolic abnormalities in peripubertal PCOS FDRs.[br]Twelve FDRs and 16 control (CON) girls aged 8-12 years, breast Tanner Stage I-III, BMI [gt]85th- [le]98th percentile were studied. Oral (OGTT, 12 FDR, 13 CON) and frequently sampled intravenous glucose tolerance testing (FSIGT, 11 FDR, 9 CON) were performed in a subgroup. Insulin sensitivity (SI), acute insulin response to glucose (AIRg), disposition index (DI) and glucose effectiveness (SG) were determined by minimal model analysis. Two-tailed t-tests or nonparametric tests were applied as appropriate, data mean[plusmn]SD.[br]Age did not differ but BMI Z-score was slightly higher in the CON (P=0.03). Three FDRs had impaired glucose tolerance (IGT); one also had impaired fasting glucose. All CON had normal glucose tolerance by design. Total testosterone (T) measured by LC-MS/MS (14[plusmn]4 FDR v 9[plusmn]8 CON ng/dL, P=0.0012) and bioavailable T (uT) (6[plusmn]3 FDR v 3[plusmn]4 CON ng/dL, P=0.0007) levels were significantly increased in FDRs. Sex hormone binding globulin, DHEAS and androstenedione levels did not differ between the groups.[br]Age and BMI Z-Score (P=0.168) were similar in the FSIGT subgroup. There was a trend toward decreased SI (1.98[plusmn]1.23 FDRs v 2.54[plusmn]1.07 CON, P=0.111) in the FDR group. AIRg and SG were similar in both groups. However, DI, the most powerful known predictor of type 2 diabetes (T2D) risk, was significantly lower in FDRs (2152[plusmn]1400 FDRs v 4026[plusmn]2249 CON, P=0.0119). These differences remained significant (P=0.027) when FDRs with dysglycemia were excluded from the analysis.[br]This is the first report of evidence for [beta]-cell dysfunction in PCOS FDR children. Further, these FDRs are among the youngest age group in whom defects in insulin secretion have been identified. These defects can occur in the absence of IGT in FDRs as they can in adult women with PCOS. T and uT levels were also increased in FDRs at earlier pubertal stages than previously reported. In contrast to prior studies, DHEAS levels did not differ in FDRs. Thus, the source of elevated T in FDRs is unclear. In conclusion, hyperandrogenemia and decreased insulin secretion are present in peripubertal PCOS FDRs.[br][br]Disclosures: NM: Principal Investigator, Pfizer, Inc., Novartis Pharmaceuticals. Nothing to Disclose: LCT, NRF, WJB, AD 2012-06-26T14:30:00 372 2012-06-26T00:00:00 1899-12-30T14:30:00 2382 403 2825 OR47-3 OR34-01 Tuesday 2383 2012


2379 ENDO12L_OR47-4 ORAL SESSION: Causes [amp] Consequences of Androgen Excess in Women (2:00 PM-3:30 PM) Obstructive Sleep Apnea in Women with Polycystic Ovary Syndrome Increases the Risk of Abnormal Glucose Tolerance Karla A Temple, Sydeaka Watson, Harry Whitmore, Eve Van Cauter, David A Ehrmann University of Chicago, Chicago, IL; University of Chicago, Chicago, IL Obstructive sleep apnea (OSA) is an independent risk factor for cardiometabolic dysfunction and is more prevalent in women with polycystic ovary syndrome (PCOS) than women without PCOS. The purpose of this study was to examine the relationships between the prevalence and severity of OSA with measures of oral glucose tolerance in women with and without OSA.[br]Following an overnight polysomnogram (PSG), 171 women (PCOS n=121; control n=50) (BMI (kg/m2): 39.1[plusmn]7.0 vs. 39.0[plusmn]7.9, p=0.9482; Age (yr): 28.3[plusmn]5.4 vs. 30.9[plusmn]6.1, p=0.0052) had a 2-h 75-g oral glucose tolerance test (OGTT) with blood sampling every 30 min for measurement of glucose and insulin concentrations. Progesterone, total testosterone, and free testosterone concentrations were measured on the fasting sample during the OGTT.[br]Eighteen (36%) control and 58 (48%) PCOS women had OSA; after controlling for BMI, age, and race, the prevalence of OSA was significantly higher in PCOS (OR=2.4 p=0.03). OSA was also more severe in women with PCOS as reflected by the Apnea-Hypopnea Index (AHI) during the PSG (median {IQR}) (2.1 {0.8-7.0} vs. 4.6 {2.3-13.8}, p= 4.7 x 10-5). Eleven (22%) control and 53 (44%) PCOS women had prediabetes (IFG or IGT) or diabetes (p[lt]0.01).[br]Logistic regression models of IGT indicated higher odds of IGT in women with PCOS after controlling for race and either OSA or AHI (OR =3.4 p= 0.01 and OR=2.9 p= 0.02 respectively). The presence and severity of OSA were determinants of IGT among women with PCOS (p[lt]0.01) but not controls (p[gt]0.16) even after controlling for age, BMI, race, as well as levels of testosterone or progesterone.[br]Two-hour glucose levels were predicted in multiple regression models controlling for age, BMI, race, and levels of testosterone or progesterone. Significant linear relationships were found in women with and without PCOS for the presence (p[lt]0.02) of OSA. In women with PCOS, a significant linear relationship was found for the severity (p[lt]0.0005) of OSA. An inverse relationship with progesterone levels and OSA severity was also observed in women with PCOS (p[lt]0.01).[br][bold]CONCLUSIONS:[/bold] Compared to controls, women with PCOS have a higher incidence of both OSA and IGT. The presence of OSA in women with PCOS increases the odds for IGT. Both severity of OSA and progesterone levels are predictive of two-hour glucose values during an OGTT in women with PCOS.[br][br]Sources of Research Support: This work was supported by NIH grants P50-HD057796, R01 HL075079, P60-DK020595, UL1-RR024999, and The Blum-Kovler Family Foundation.[br][br]Nothing to Disclose: KAT, SW, HW, EVC, DAE 2012-06-26T14:45:00 372 2012-06-26T00:00:00 1899-12-30T14:45:00 552 403 2826 OR47-4 OR34-01 Tuesday 2384 2012


2380 ENDO12L_OR47-5 ORAL SESSION: Causes [amp] Consequences of Androgen Excess in Women (2:00 PM-3:30 PM) Coronary Endothelial Dysfunction and Cardiovascular Performance in Polycystic Ovarian Syndrome and Their Relationship to Testosterone and Insulin Sensitivity Alice Y Chang, Samantha Thompson, Peter Snell, M Dean Palmer, Kara Boyd, Richard J Auchus, Benjamin D Levine University of Texas Southwestern Medical Center, Dallas, TX; Institute for Exercise and Environmental Medicine, Dallas, TX; University of Michigan, Ann Arbor, MI Polycystic Ovarian Syndrome (PCOS) is a condition of androgen excess associated with impaired insulin sensitivity. Although PCOS has been associated with peripheral endothelial dysfunction at rest and lower peak oxygen uptake (VO[sub]2max[/sub]) compared to healthy controls, we sought to determine if this was also true compared to women who have the metabolic syndrome (MetS) but not PCOS. In addition, we specifically examined coronary endothelial function and measurements of stroke volume (SV) and cardiac output (CO) during stress.[br][bold]Methods.[/bold] We recruited premenopausal, obese women with PCOS (n=18) or MetS (n=19) matched for age (all [ge]30 yrs) and ethnicity. Percent body fat was calculated from underwater weights. Coronary flow was measured with 3T MRI (spiral, velocity-encoded sequences of the right coronary artery) during a 2 minute cold pressor test. Left ventricular mass was measured from short axis 8 mm slices using semi-automated MASS software. During an exercise treadmill test, VO[sub]2max[/sub] and cardiac output measurements were obtained using Douglas bags and the acetylene rebreathing technique. Stroke volume was calculated from cardiac output and heart rate. Insulin sensitivity was calculated from measurements during a frequently sampled intravenous glucose tolerance test.[br][bold]Results.[/bold] There were no differences between the PCOS or MetS groups in age, body mass index, ethnicity or percent body fat. Compared to women with MetS, women with PCOS had significantly higher total testosterone (PCOS: 45.0[plusmn]25; MetS: 29.9[plusmn]12.7 ng/dL) and lower insulin sensitivity (PCOS: 1.7[plusmn]0.9; MetS: 3.1[plusmn]1.2 (mU/L)[sup]-1[/sup]min[sup]-1[/sup], p[lt]0.01). During the 2[sup]nd[/sup] minute of the cold pressor test, when endothelial-mediated vasodilatation should occur, PCOS was associated with a significant decrease in coronary flow (PCOS: -26.0[plusmn]38.1; MetS: 41.2[plusmn]30.1%, p=0.03). There was no significant difference in left ventricular mass indexed to height. During exercise, there was no significant difference in VO[sub]2max[/sub], CO or SV indexed to body surface area. When analyzing the women together as one group, indexed SV was significantly associated with insulin sensitivity (r=0.6, p[lt]0.01) but not testosterone.[br][bold]Conclusions.[/bold] Women with PCOS have greater impairment in coronary endothelial function and insulin sensitivity than obese women with the metabolic syndrome. During exercise, there were no significant differences in VO[sub]2max[/sub], indexed stroke volume or cardiac output, but rather an association of indexed SV with insulin sensitivity.[br][br]Sources of Research Support: American Heart Association Fellow to Faculty Award.[br][br]Disclosures: RJA: Investigator, Jansen Pharmaceuticals. Nothing to Disclose: AYC, ST, PS, MDP, KB, BDL 2012-06-26T15:00:00 372 2012-06-26T00:00:00 1899-12-30T15:00:00 1311 403 2827 OR47-5 OR34-01 Tuesday 2385 2012


2381 ENDO12L_OR47-6 ORAL SESSION: Causes [amp] Consequences of Androgen Excess in Women (2:00 PM-3:30 PM) Impact of Glucocorticoid Receptor Gene Polymorphisms on Metabolic Profile of Adult Patients with Classical Form of 21-Hydroxylase Deficiency (CAH) Ricardo PP Moreira, Larissa G Gomes, Berenice Bilharinho Mendonca, Tania ASS Bachega Faculdade de Medicina da Universidade de S[atilde]o Paulo, S[atilde]o Paulo, Brazil Recent evidences suggest that adult CAH patients have an increased risk for cardiovascular disease and it remains unknown if lifelong glucocorticoid (GC) treatment could contribute to this risk. In the general population, some glucocorticoid receptor gene (GR) polymorphisms are associated with hypertension, obesity and dyslipidemia; however, their influence on metabolic profile of CAH patients has not been evaluated. [bold]Objectives: [/bold]To analyze the association between GR polymorphisms and metabolic profile in CAH patients. [bold]Methods:[/bold] 68 adult patients (40 females, 34SV, 34SW) at a mean age of 28.4 [plusmn]9 years receiving dexamethasone (mean 0.27 [plusmn] 0.11mg/day) to obtain normal androgen levels. SW patients also received fludrocortisone (50 mcg/day). MetS and obesity were defined by NCEP ATPIII criteria and by BMI [gt]30kg/m[sup2], respectively. The N363S, ER22/23EK and A3669G alleles were genotyped by sequencing and the [italic]Bcl[/italic]I by allelic-specific PCR. Association analysis among GR polymorphisms and metabolic profile were carried out with Chi-square, [italic]t[/italic]-test and regression analysis. [bold]Results: [/bold]Obesity and MetS were observed in 21% and 9% of patients, respectively, and were not correlated with GC doses and duration of therapy. BMI was positively correlated with higher blood pressure (BP), TG and LDL-c levels and HOMA-IR ([italic]P[/italic][lt]0.01) and inversely correlated with HDL-c levels ([italic]P[/italic]=0.03). The [italic]Bcl[/italic]I and A3669G variants were found, in Hardy-Weinberg equilibrium, in 26.4% and 9.6% of alleles, respectively. The N363S and ER22/23EK alleles were identified in only 1 patient, each one. [italic]Bcl[/italic]I carriers presented higher BMI (29kg/m[sup2] [plusmn]5.3 [italic]vs[/italic] 26kg/m[sup2] [plusmn]5.3, respectively[italic] P=[/italic]0.02) and higher waist circumference (89cm [plusmn]12.7 [italic]vs[/italic] 81cm [plusmn]13, [italic]P=[/italic]0.04, respectively) compared to wild type carriers. Hypertension was found in 12% of patients and [italic]Bcl[/italic]I carriers presented higher systolic BP than non-carriers (125mmHg [plusmn]7 [italic]vs[/italic] 117mmHg [plusmn]9, [italic]P=[/italic]0.008, respectively). Low HDL-c and high triglycerides (TG) levels were identified in 10% and 30% of patients, respectively, and were not associated with GR variants. Fasting glucose [ge]110mg/dL was not observed. A3669G carriers and non-carriers did not differ in the metabolic profile. [bold]Conclusion:[/bold] This is the first study evaluating the impact of GR polymorphisms on adverse metabolic profile in CAH patients. There is a significant association between [italic]Bcl[/italic]I polymorphism, which increases GC sensitivity, with obesity and hypertension in CAH. Genotyping GR variants may benefit long-term outcome.[br][br]Nothing to Disclose: RPPM, LGG, BBM, TASSB 2012-06-26T15:15:00 372 2012-06-26T00:00:00 1899-12-30T15:15:00 1455 403 2828 OR47-6 OR34-01 Tuesday 2386 2012


2382 ENDO12L_OR48-1 ORAL SESSION: Endocrine Hypertension: The Renin-Angiotensin-Aldosterone System (2:00 PM-3:30 PM) Autonomic Autoantibodies with Allosteric Activity in Idiopathic Postural Orthostatic Hypotension (IOH) and Tachycardia Syndrome (POTS): A New Mechanism Hongliang Li, Muneer Ahmad Khan, Megan Daniel Vanderlinde-Wood, Allison Lea Galloway, Caitlin Marie Zillner, Xichun Yu, David Charles Kem University of Oklahoma and Veterans Adminstration Medical Centers, Oklahoma City, OK [bold]Objective:[/bold] Patents with postural orthostatic tachycardia syndrome (POTS) have a variable drop in BP that is partially or completely compensated by the exaggerated tachycardia. We have reported circulating agonistic autoantibodies to the [beta]2 adrenergic receptor ([beta]2AR) and M3 muscarinic receptor (M3R) serve as vasodilators and cause or exacerbate orthostatic hypotension. Autoantibodies to the [beta]1 adrenergic receptor ([beta]1AR), M2 muscarinic receptor (M2R) and [alpha]1 adrenergic receptor ([alpha]1AR) are variably co-present and may explain the pathophysiology of this complex syndrome.[br][bold]Methods:[/bold] Patients were categorized into 16 with idiopathic orthostatic hypotension (IOH), 7 with POTS and 10 normal control subjects. Purified IgG from all patients and healthy control subjects were examined in a receptor-transfected cell-based assay for [beta]1AR and [beta]2AR activation, and a [beta]-arrestin-based assay for M3R activation. Data are expressed as % of baseline (Mean [plusmn] SD). All subjects gave informed consent.[br][bold]Results:[/bold] Patients with IOH and POTS had increased [beta]2AR activity (122 [plusmn] 14% and 121 [plusmn] 12%) compared to normal subjects (106 [plusmn] 8%, P=0.007). [beta]1AR activity was similarly increased in both IOH and POTS compared to normal controls. By contrast, M3R activation in IOH was increased (141 [plusmn] 32%) over normal controls (99.4 [plusmn] 12 %, P=0.002) while activity for POTS M3R was not significantly increased (114 [plusmn] 30%). IOH and POTS had similar M2R inhibition of forskolin-induced cAMP production (72 [plusmn] 16% and 76 [plusmn] 12%) compared to normal controls (97 [plusmn] 4%). We have demonstrated the vasodilatory capacity of the [beta]2AR and M3R autoantibodies in vitro using isolated perfused rat cremaster arterioles. Several subjects had increased ELISA evidence for autoantibodies to [alpha]1AR.[br][bold]Discussion:[/bold] Vasodilatory autoantibodies to [beta]2AR and M3R are present in IOH patients suggesting these autoantibodies cause or exacerbate orthostasis by altering the compensatory postural vascular response. Patients with POTS also demonstrate a high proportion of activating autoantibodies and may represent a subset with autoantibodies to the [alpha]1AR that are partially inhibitory to the native norepinephrine ligand in vivo.[br][bold]Conclusion:[/bold] These data support the concept that circulating agonistic autoantibodies are present and active in IOH and in POTS. POTS may represent one end of a spectrum of autoimmune disease with the potential for autoimmune allosteric inhibition of norepinephrine action in [alpha]1AR but not in [beta]1/2AR.[br][br](1) Yu X et al., JASH 2012;Jan-Feb;6(1):40. (2) Li H et al., Hypertension 2012;Feb;59(2):402.[br][br]Nothing to Disclose: HL, MAK, MDV-W, ALG, CMZ, XY, DCK 2012-06-26T14:00:00 310 2012-06-26T00:00:00 1899-12-30T14:00:00 1875 404 2829 OR48-1 OR09-01 Tuesday 2387 2012


2383 ENDO12L_OR48-2 ORAL SESSION: Endocrine Hypertension: The Renin-Angiotensin-Aldosterone System (2:00 PM-3:30 PM) Protein Kinase C Beta Plays an Important Role in High Glucose-Induced Stabilization of Mineralocorticoid Receptor Takeshi Hayashi, Hirotaka Shibata, Isao Kurihara, Ayano Murai-Takeda, Yuko Mitsuishi, Rie Jo, Takako Ohyama, Hiroshi Itoh Keio University, School of Medicine, Tokyo, Japan [Aim] Antihypertensive efficacy of mineralocorticoid receptor (MR) antagonist is shown in patients with primary aldosteronism. Several subsets of resistant hypertension, including diabetes mellitus can also be effectively controlled by add-on therapy with MR antagonist. These findings indicate that MR is overactivated in selected diabetic patients even in the absence of high plasma aldosterone levels. One of the adverse effects of hyperglycemia is the chronic activation of protein kinase C (PKC). We investigated whether and how high glucose affects MR activation through PKC signaling.[br][Materials and Methods] We utilized COS-7 and HEK293 cells. The MR transcriptional activities were evaluated with reporter-based transient transfection assays. The levels of expression of MR were examined by Western blotting. The MR ubiquitylation was examined by coimmunoprecipitation assays. High and normal glucose media contained 30mM and 5.6mM glucose, respectively.[br][Results] First, transient transfection assays in COS-7 cells showed that 10[sup]-10[/sup]M aldosterone increased MR transactivation by 100-fold. High glucose treatment (30mM) enhanced the MR transactivation by 2-fold compared with normal glucose treatment (5.6mM), and the activities were attenuated by introduction of siRNA for PKCbeta, but not PKCalpha. These data indicate that high glucose enhances MR transactivation mainly via PKCbeta activation. Next, we investigated levels of expression of MR protein. Under high glucose conditions, levels of MR protein were significantly increased by 2-fold in a time-dependent manner. The MR levels were decreased by introduction of siRNA for PKCbeta, but not PKCalpha, indicating that high glucose activates PKCbeta, thus resulting in increased MR protein levels. Finally, we investigated the ubiquitylation of MR protein with coimmunoprecipitation assays. The MR was significantly associated with ubiquitin in the presence of aldosterone; however, high glucose condition inhibited the MR ubiquitylation, thus resulting in increase in MR protein levels.[br][Conclusion] These data suggest that high glucose condition activates several PKC isoforms including PKCbeta, thus resulting in enhancement of aldosterone-mediated MR transactivation via MR protein stabilization in vitro. The results may support the clinical observation that MR antagonist has renoprotective effects in diabetic patients with normal aldosterone levels.[br][br]Nothing to Disclose: TH, HS, IK, AM-T, YM, RJ, TO, HI 2012-06-26T14:15:00 310 2012-06-26T00:00:00 1899-12-30T14:15:00 1326 404 2830 OR48-2 OR09-01 Tuesday 2388 2012


2384 ENDO12L_OR48-3 ORAL SESSION: Endocrine Hypertension: The Renin-Angiotensin-Aldosterone System (2:00 PM-3:30 PM) Telmisartan Stimulates Aldosterone Synthase Gene ([italic]CYP11B2[/italic]) Expression Via the CaMK-Mediated Induction of NURR1 Ken Matsuda, Akira Uruno, Takako Saito-Ito, Kyoko Shimizu, Masataka Kudo, Fumitoshi Satoh, Takeo Yoshikawa, Akiko Saito-Hakoda, Sadayoshi Ito, Akira Sugawara Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku University Graduate School of Medicine, Sendai, Japan; Tohoku University Graduate School of Medicine, Sendai, Japan Introduction: Aldosterone synthase gene ([italic]CYP11B2[/italic]) encodes the key enzyme of adrenal aldosterone production. In ENDO 2011, we demonstrated that telmisartan, but not other angiotensin (A) II receptor blockers, stimulate [italic]CYP11B2[/italic] transcription/mRNA expression and aldosterone secretion in the absence of AII using human adrenal H295R cells. We therefore examined the mechanisms of the telmisartan-mediated [italic]CYP11B2[/italic] transcription using a newly obtained stable H295R cell line expressing [italic]CYP11B2[/italic] 5[apos]-flanking region/luciferase cDNA chimeric construct. Methods: A stable H295R cell line expressing [italic]CYP11B2[/italic] 5[apos]-flanking region/luciferase cDNA chimeric construct was obtained after the selection by hygromycin B. In some experiments, transient transfection studies were also performed. mRNA expression of [italic]CYP11B2[/italic] and Nur-related factor 1 (NURR1) in H295R cells was determined by real-time PCR. Results: Telmisartan increased [italic]CYP11B2[/italic] transcription/mRNA expression dose-dependently and maximally at 12 h. Transient transfection experiments using [italic]CYP11B2[/italic] 5[apos]-flanking region deletion mutants/Ad5 point mutant indicated that the Ad5 element was responsible for the telmisartan effect. Examinations of transcription factors known to bind to the Ad5 element revealed that telmisartan significantly induced NURR1 expression maximal at 1 h. The telmisartan effect on [italic]CYP11B2[/italic] transcription/mRNA expression was abrogated by KN-93, a Ca[sup]2+[/sup]/calmodulin-dependent kinase (CaMK) inhibitor. The telmisartan-induced NURR1 mRNA expression was also abrogated by KN-93. High-dose olmesartan treatment did not affect the telmisartan-mediated [italic]CYP11B2[/italic] transcription. Conclusion: Telmisartan may stimulate [italic]CYP11B2[/italic] transcription via the CaMK-mediated induction of NURR1 that activate the Ad5 element, and AII type 1 receptor may not be involved in the effect.[br][br]Nothing to Disclose: KM, AU, TS-I, KS, MK, FS, TY, AS-H, SI, AS 2012-06-26T14:30:00 310 2012-06-26T00:00:00 1899-12-30T14:30:00 1461 404 2831 OR48-3 OR09-01 Tuesday 2389 2012


2385 ENDO12L_OR48-4 ORAL SESSION: Endocrine Hypertension: The Renin-Angiotensin-Aldosterone System (2:00 PM-3:30 PM) Demonstration of Blood Pressure-Independent Non-Ischemic Myocardial Fibrosis in Primary Aldosteronism [mdash] A Cardiac MRI Study E Marie Freel, Patrick B Mark, Robin AP Weir, Emily McQuarrie, Karen Allan, Alan G Jardine, Eleanor Davies, John MC Connell University of Glasgow, Glasgow, UK; Hairmyres Hospital, East Kilbride, UK; University of Dundee, Dundee, UK [underline]Background:[/underline] Primary Aldosteronism (PA) is common and associates with excess cardiovascular morbidity independent of blood pressure. It has been demonstrated that exposure to aldosterone and sodium leads to cardiac fibrosis and hypertrophy in humans and animals and this may be mediated by inflammation and oxidative stress. We aimed to clarify the effects of aldosterone excess on myocardial structure and composition in human subjects using contrast-enhanced cardiac MRI as well as explore potential underlying mechanisms for any observed differences.[br][underline]Methods:[/underline] Twenty seven subjects with recently diagnosed (less than one year) PA and 54 matched essential hypertensive (EH) controls underwent gadolinium-enhanced cardiac MRI; non-infarct related myocardial fibrosis was identified by a diffuse pattern of late gadolinium enhancement (LGE). Patients also underwent assessment of pulse wave velocity (PWV) by applanation tonometry, measurement of circulating superoxide anion and C-reactive protein as well as resting blood pressure and 24h urinary sodium.[br][underline]Results:[/underline] Subjects were well matched with no difference in severity nor duration of hypertension. There was a significant increase in the frequency of diffuse LGE in PA (67%) when compared to EH subjects (17%; p[lt]0.0001) although no difference in LV mass or function. This finding was independent of blood pressure and multivariate logistic regression modelling confirmed the presence of aldosterone excess as the only independent predictor of myocardial fibrosis (p[lt]0.01). Carotid-femoral PWV as well as superoxide and CRP were significantly higher in PA subjects.[br][underline]Conclusions:[/underline] These data demonstrate that PA patients demonstrate more frequent myocardial fibrosis; this finding is independent of blood pressure. This may be mediated in part through inflammation and oxidative stress. This study highlights the importance of specific targeting of aldosterone excess as well as blood pressure reduction to minimise cardiac morbidity in Primary Aldosteronism.[br][br]Sources of Research Support: Medical Research Council (UK) Clinician Scientist Fellowship to Dr Freel.[br][br]Nothing to Disclose: EMF, PBM, RAPW, EM, KA, AGJ, ED, JMCC 2012-06-26T14:45:00 310 2012-06-26T00:00:00 1899-12-30T14:45:00 170 404 2832 OR48-4 OR09-01 Tuesday 2390 2012


2386 ENDO12L_OR48-5 ORAL SESSION: Endocrine Hypertension: The Renin-Angiotensin-Aldosterone System (2:00 PM-3:30 PM) Role of microRNA-21 in Aldosterone-Mediated Cardiac Injury Jana E Price, Michael E Hall, Licy L Yanes, Damian G Romero University of Mississippi Medical Center, Jackson, MS; University of Mississippi Medical Center, Jackson, MS Primary hyperaldosteronism is characterized by excess autonomous secretion of aldosterone (ALDO) independent of the renin-angiotensin system and accounts for [sim]10% of hypertensive patients. Excess ALDO, inappropriate for the salt intake status, causes cardiac hypertrophy, inflammation, fibrosis and hypertension. The molecular mechanisms that trigger the onset and progression of ALDO-mediated cardiac injury are poorly understood.[br]MicroRNAs (miRNAs) are endogenous, small, non-coding RNAs that have important roles in development and cell proliferation and differentiation. miRNAs downregulate the expression levels of specific proteins by translational repression or mRNA degradation. Several miRNAs have been implicated in diverse cardiac pathologies in humans and experimental models.[br]To study if miRNAs are involved in ALDO/SALT-mediated cardiac injury, we performed a time-series analysis of left ventricle (LV) miRNA expression. Uninephrectomized male Sprague Dawley rats were treated with ALDO (0.75 [micro]g/h) and 1.0% NaCl/0.3% KCl in the drinking water for 2, 7, 14, 28 or 56 days. miRNA microarrays followed by qRT-PCR showed that ALDO/SALT time-dependently modulate the expression of 13 miRNAs in the LV. miR-21 increased 3.8-fold after 2 weeks of treatment and remained elevated for the rest of the study. To study the role or miR-21 in ALDO/SALT-mediated cardiac injury, ALDO/SALT-treated rats were treated with miR-21 antagomirs 7 days before the end of the 2 week treatment period. miR-21 downregulation with antagomirs (confirmed by qPCR and Northern-blot) increased LV collagen I, collagen III and fibronectin mRNA expression. LV ALDO/SALT-mediated miR-21 upregulation occurs mainly in cardiac fibroblasts as determined by in situ hybridization and miR-21 quantification in isolated cardiomyocyte/non-cardiomyocyte fractions.[br]AntagomiR-21 exacerbated ALDO/SALT-mediated cardiac hypertrophy as determined by an increase in LV weight/tibia length ratio, which correlates with an increase in end-diastolic posterior and anterior wall thickness. Assessment of cardiac function by echocardiography showed that antagomiR-21 treatment exacerbated ALDO/SALT attenuation of LV systolic function as observed by significant decreases in cardiac output and fractional shortening.[br]In summary, miR-21 is upregulated by ALDO/SALT treatment in LV fibroblasts and may function as a compensatory mechanism that mitigates ALDO/SALT-mediated deleterious cardiac effects.[br][br]Sources of Research Support: American Heart Association Grant 12SDG8980032 awarded to DGR.[br][br]Nothing to Disclose: JEP, MEH, LLY, DGR 2012-06-26T15:00:00 310 2012-06-26T00:00:00 1899-12-30T15:00:00 1708 404 2833 OR48-5 OR09-01 Tuesday 2391 2012


2387 ENDO12L_OR48-6 ORAL SESSION: Endocrine Hypertension: The Renin-Angiotensin-Aldosterone System (2:00 PM-3:30 PM) Spontaneous Remission of Primary Aldosteronism after Long-Term Treatment with Mineralocorticoid Receptor Antagonists Takashi Yoneda, Mitsuhiro Kometani, Masashi Demura, Shigehiro Karashima, Nobushige Oda, Shunsuke Mori, Masashi Oe, Atsushi Hashimoto, Toshitaka Sawamura, Mikiya Usukura, Masakazu Yamagishi, Yoshiyu Takeda Graduate School of Medical Science, Kanazawa University, Kanazawa, Japan (Context) Primary aldosteronism (PA) is mainly caused by aldosterone producing adenoma and idiopathic bilateral adrenal hyperplasia (IHA). Laparoscopic adrenalectomy is recommended for the treatment of APA. Medical treatment with a mineralocorticoid receptor (MR) antagonist such as spironolactone (SP) or eplerenone(EP) is recommended for patients with IHA. Armani et al reported potassium canrenoate or SP therapy has produced spontaneous remission of IHA (J Endocrinol Invest 2005, Hypertension 2007). Fishcher et al reported that the prevalence of spontaneous remission of IHA was 5.4%(Clin Endocrinol 2011). Moreover we reported a case of APA with spontaneous remission after long-term SP therapy (JCEM 2012 in press).[br](Objective) We aimed to determine the prevalence of spontaneous remission of PA during long-term treatment with MR antagonists (SP or EP).[br](Methods) 56 patients with PA (APA: 9 cases, IHA: 26 cases, Subtype unknown: 21 cases) treated with MR antagonists during more than 3 years were investigated. 36 patients were treated with SP. 20 patients were treated with EP. The patients were identified retrospectively by chart review and prospectively assessed by clinical and biochemical means. We defined complete remission (CR) of PA as normal aldosterone to renin ratio (ARR), normal suppression test, normalization of hypokalemia without hypertension. Partial remission (PR) was defined as normalization of normal ARR, normal suppression test, normalization of hypokalemia with hypertension.[br](Results)The mean period of MR antagonist treatment was 4.1 years in the patients. We identified 2 of 56 (3.6%) patients with CR, one with remission of APA treated with SP, and the other with remission of IHA treated with SP. We also identified 8 of 56 (14.3%) patients with PR, three with remission of IHA and one with remission of subtype unknown treated with SP, two with remission of IHA and two with remission of subtypes unknown treated with EP.[br](Conclusion) Remission of PA may occur in some patients after long-term MR treatment.[br][br]Nothing to Disclose: TY, MK, MD, SK, NO, SM, MO, AH, TS, MU, MY, YT 2012-06-26T15:15:00 310 2012-06-26T00:00:00 1899-12-30T15:15:00 1513 404 2834 OR48-6 OR09-01 Tuesday 2392 2012


2388 ENDO12L_OR49-1 ORAL SESSION: Novel Treatment Approaches for Pituitary Tumors (2:00 PM-3:30 PM) Patients with Cushing Disease Achieve Normal Urinary Cortisol with LCI699, a Potent 11[beta]-Hydroxylase Inhibitor: Preliminary Results from a Multicenter, Proof-of-Concept Study Rosario Pivonello, Maria Fleseriu, Laurence Guignat, Yiming Zhang, Paul Robinson, Ann Taylor, Catherine Watson, Mario Maldonado, Amir H Hamrahian, Marco Boscaro, Beverly MK Biller University of Naples [apos]Federico II[apos], Naples, Italy; Oregon Health [amp] Science University, Portland, OR; Cochin Hospital, Paris, France; Universit[eacute] Paris 5, Paris, France; Novartis Pharmaceuticals Corporation, East Hanover, NJ; Novartis Pharmaceuticals UK Limited, Horsham, UK; Novartis Institutes for BioMedical Research, Cambridge, MA; Novartis Pharma AG, Basel, Switzerland; Cleveland Clinic Foundation, Cleveland, OH; Polytechnic University of Marche, Ancona, Italy; Massachusetts General Hospital, Boston, MA [bold]Introduction:[/bold] Endogenous Cushing[apos]s syndrome is caused by chronic elevation of circulating cortisol levels and causes many clinical morbidities. LCI699 is a potent inhibitor of 11[beta]-hydroxylase, the enzyme which catalyzes the final step of cortisol synthesis. LCI699 may thus be a potential new treatment for all forms of Cushing[apos]s syndrome. A proof-of-concept study was performed to evaluate whether LCI699 can reduce cortisol levels, as evaluated by 24-hour urinary free cortisol (UFC).[br][bold]Methods:[/bold] Adult patients with mild-to-severe Cushing[apos]s disease (UFC[gt]1.5x the upper limit of normal [ULN], mean of three collections in 14 days) received oral LCI699 for 10 weeks in an open-label study. LCI699 was initiated at 2mg bid. Dose escalation was planned every 2 weeks to 5, 10, 20 and 50mg bid until UFC normalized, in which case the dose was maintained until day 70, when treatment stopped. Dose reduction due to cortisol withdrawal symptoms or intolerance was permitted. UFC was assessed on the penultimate day of each 2-week period. Patients were monitored until day 84. The primary endpoint was UFC[le]ULN or a [ge]50% decrease from baseline at day 70 using the mean of three UFC samples collected during the week before day 70.[br][bold]Results:[/bold] Eleven patients (aged 25[ndash]55 years; 4 men) were enrolled and completed the study. Baseline UFC range was 1.6[ndash]17.0xULN. UFC levels were normal on at least one post-treatment assessment in all 11 patients. The primary endpoint was achieved by all 11 patients who completed the active treatment phase, 10 of whom had normal UFC levels on day 70. After treatment discontinuation, UFC was [gt]ULN in all 9 patients with measurements available at day 84 at the time of abstract submission. The median dose of LCI699 associated with UFC normalization was between 5 and 10mg bid. Mean SBP and DBP levels were reduced by 13.1 and 9.3mmHg, respectively, compared with baseline. Levels of 11-deoxycortisol tended to increase during treatment and decline after day 70. LCI699 was generally well tolerated; the most frequently reported adverse events were fatigue (5/11), nausea (4/11) and headache (3/11). Five of the 11 patients experienced ACTH levels [gt]2xbaseline. Four patients experienced study drug-related hypokalemia (K[sup]+[/sup][lt]3.5mmol/L; min 3.1mmol/L). There were no serious AEs of suspected drug relationship.[br][bold]Conclusion: [/bold]LCI699 demonstrated efficacy with a satisfactory safety profile in this short-term proof-of-concept study in patients with Cushing[apos]s disease.[br][br]Sources of Research Support: Novartis Pharmaceuticals.[br][br]Disclosures: RP: Consultant, Novartis Pharmaceuticals. MF: Ad Hoc Consultant, Novartis Pharmaceuticals, Ipsen. YZ: Employee, Novartis Pharmaceuticals. PR: Employee, Novartis Pharmaceuticals. AT: Employee, Novartis Pharmaceuticals. CW: Employee, Novartis Pharmaceuticals. MM: Employee, Novartis Pharmaceuticals. AHH: Speaker, Novartis Pharmaceuticals. BMKB: Consultant, Novartis Pharmaceuticals, Corcept Therapeutics. Nothing to Disclose: LG, MB 2012-06-26T14:00:00 Theater C 2012-06-26T00:00:00 1899-12-30T14:00:00 1892 405 2835 OR49-1 OR22-02 Tuesday 2393 2012


2389 ENDO12L_OR49-2 ORAL SESSION: Novel Treatment Approaches for Pituitary Tumors (2:00 PM-3:30 PM) Initial Response to Pasireotide Treatment Is Predictive of 12-Month Response: Results of a Large, Randomized, Double-Blind, Phase III Study in Patients with Cushing Disease Andre Lacroix, William Ludlam, Franco Mantero, Abdurrahman Comlekci, Mauro Czepielewski, Mario Maldonado, Gareth Hughes, Andrew Trovato, Antoine Tabarin, Marco Boscaro Centre Hospitalier de l[apos]Universit[eacute] de Montr[eacute]al, Montr[eacute]al, Canada; Swedish Neuroscience Institute, Seattle, WA; University of Padua, Padua, Italy; Dokuz Eylul University School, Izmir, Turkey; Hospital de Cl[iacute]nicas de Porto Alegre, Porto Alegre, Brazil; Novartis Pharma AG, Basel, Switzerland; Novartis Pharmaceuticals Corporation, East Hanover, NJ; Universit[eacute] Bordeaux 2, Centre Hospitalier Universitaire de Bordeaux, Bordeaux, France; Polytechnic University of Marche, Ancona, Italy [bold]Introduction: [/bold]Patients with uncontrolled Cushing[apos]s disease have increased morbidity and mortality; rapid and sustained correction of their hypercortisolism is important. Pasireotide can lead to reductions in UFC levels and improvements in symptoms of Cushing[apos]s disease. The analyses presented here evaluate the predictive value of early response.[br][bold]Methods: [/bold]Patients (n=162; 126 female; mean age 40.2 yrs) with persistent/recurrent or [italic]de novo[/italic] (if not surgical candidates) Cushing[apos]s disease and UFC levels [ge]1.5x the upper limit of normal (ULN) were randomized to pasireotide 600[mu]g (n=82) or 900[mu]g (n=80) sc bid. Dose titration (max: 1200[micro]g bid) was allowed after month 3. UFC control was defined as UFC[le]ULN, partial control as UFC[gt]ULN and [ge]50% reduction from baseline, and uncontrolled UFC as UFC[gt]ULN without a [ge]50% reduction from baseline.[br][bold]Results:[/bold] Overall, 90 (55.6%) patients had controlled/partially controlled UFC levels at month 1 or 2, and 72 (44.4%) patients had uncontrolled UFC levels at months 1 and 2. Of the patients with controlled/partially controlled UFC at month 1 or 2, 47 (52.2%) and 38 (42.2%) were controlled/partially controlled at months 6 and 12, respectively. Of those with uncontrolled UFC at months 1 and 2, 66 (91.7%) and 64 (88.9%) were uncontrolled at months 6 and 12. Of the patients with fully/partially controlled UFC levels at month 1 or 2, 61 (67.8%) remained on treatment until month 12, compared with 17 (23.6%) patients whose UFC levels were uncontrolled at months 1 and 2. Patients whose UFC levels were controlled or partially controlled at month 1 or 2 had significant decreases in BMI, weight, total cholesterol, triglycerides, and systolic and diastolic blood pressure between baseline and month 12. Patients with uncontrolled UFC at months 1 and 2 had significant decreases only in BMI and weight. Pasireotide had a similar safety profile to other somatostatin analogues, except for hyperglycemia-related events (72.8%); 84.7% of patients who had a hyperglycemia-related event did so by month 3.[br][bold]Conclusion: [/bold]Pasireotide leads to rapid and sustained decreases in UFC in the majority of patients. Most patients who have an early response to pasireotide stay on treatment and the majority of these continue to respond. In addition, the majority of patients who do not respond to pasireotide within the first 2 months do not respond at a later time point. Treatment decisions in patients taking pasireotide can therefore be made early in the therapeutic course.[br][br]Sources of Research Support: Novartis Pharmaceuticals.[br][br]Disclosures: AL: Investigator, Novartis Pharmaceuticals; Speaker, Novartis Pharmaceuticals. WL: Speaker, Pfizer, Inc., Novartis Pharmaceuticals, Ipsen; Medical Advisory Board Member, Corcept Therapeutics, Novartis Pharmaceuticals, Ipsen, Endo Pharmaceuticals Inc. MM: Employee, Novartis Pharmaceuticals. GH: Employee, Novartis Pharmaceuticals. AT: Employee, Novartis Pharmaceuticals. AT: Consultant, Novartis Pharmaceuticals. Nothing to Disclose: FM, AC, MC, MB 2012-06-26T14:15:00 Theater C 2012-06-26T00:00:00 1899-12-30T14:15:00 1718 405 2836 OR49-2 OR22-02 Tuesday 2394 2012


2390 ENDO12L_OR49-3 ORAL SESSION: Novel Treatment Approaches for Pituitary Tumors (2:00 PM-3:30 PM) Pasireotide LAR Is Significantly More Effective Than Octreotide LAR at Providing Biochemical Control in Patients with Acromegaly: Results of a 12-Month Randomized, Double-Blind, Multicenter, Phase III Study Marcello D Bronstein, Michael Sheppard, Pamela Freda, Feng Gu, Chiung-Chyi Shen, Monica Gadelha, Maria Fleseriu, Karina Hermosillo Resendiz, Matthieu Ruffin, Yinmiao Chen, Annamaria Colao University of S[atilde]o Paulo Medical School, S[atilde]o Paulo, Brazil; University of Birmingham, Edgbaston, Birmingham, UK; Columbia University College of Physicians [amp] Surgeons, New York, NY; Peking Union Medical College Hospital, Beijing, China; National Yang-Ming University, Taipei, Taiwan; Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; Oregon Health [amp] Science University, Portland, OR; Novartis Pharmaceuticals Corporation, Florham Park, NJ; Novartis Pharma AG, Basel, Switzerland; Novartis Pharmaceuticals Corporation, Florham Park, NJ; University of Naples [apos]Federico II[apos], Naples, Italy [bold]Introduction:[/bold] Using GH[lt]2.5[mu]g/L [italic]and[/italic] normalized IGF-1, response rates are 20[ndash]25% after 12m with currently available somatostatin analogues in medically na[iuml]ve patients with acromegaly. The broader somatostatin receptor binding profile of pasireotide may potentially improve response rates. This randomized, double-blind, 12m study compared pasireotide LAR with octreotide LAR.[br][bold]Methods:[/bold] Patients with acromegaly (GH[gt]5[micro]g/L or GH nadir[ge]1[micro]g/L post-OGTT, and IGF-1[gt]ULN) who were [italic]de novo[/italic] with a visible adenoma on MRI or medically na[iuml]ve (no previous medical therapy but prior pituitary surgery) received pasireotide LAR 40mg/28d (n=176) or octreotide LAR 20mg/28d (n=182) for 12m. At 3 and 7m, titration to pasireotide LAR 60mg or octreotide LAR 30mg for suboptimal biochemical response was permitted, but not mandatory. Primary objective: comparison of the proportion of patients in each arm with GH[lt]2.5[micro]g/L [italic]and [/italic]normal IGF-1 at 12m.[br][bold]Results:[/bold] At baseline, mean GH was 21.9 and 18.8[mu]g/L in the pasireotide LAR and octreotide LAR arms, respectively; mean IGF-1 was 2.6xULN and 2.8xULN. 80.1% and 85.7% of pasireotide LAR and octreotide LAR patients completed 12m; 50.6% and 67.6% were uptitrated. Mean GH and IGF-1 decreased within 3m and remained suppressed. Pasireotide LAR provided superior biochemical control compared with octreotide LAR; 31.3% vs 19.2% of patients achieved GH[lt]2.5[mu]g/L [italic]and[/italic] normalized IGF-1 ([italic]P[/italic]=0.007). Pasireotide LAR therapy was 63% more likely to provide full biochemical control than octreotide LAR. Normal IGF-1 was reported in 38.6% and 23.6% ([italic]P[/italic]=0.002) of patients, and GH[lt]2.5[micro]g/L was reported in 48.3% and 51.6% ([italic]P[/italic]=0.536) of patients receiving pasireotide LAR and octreotide LAR, respectively. The most common AEs with pasireotide LAR vs octreotide LAR were diarrhea (39.3 vs 45.0%), cholelithiasis (25.8 vs 35.6%) and headache (18.5 vs 26.1%). All hyperglycemia-related AE terms were grouped; hyperglycemia-related AEs were seen in 57.3% and 21.7% of patients. Most AEs were mild or moderate.[br][bold]Conclusions:[/bold] In the largest randomized study of a medical therapy in patients with acromegaly, pasireotide LAR was superior to octreotide LAR in providing full biochemical control. Apart from the frequency and severity of hyperglycemia, the safety profile of pasireotide LAR was similar to that of octreotide LAR. These results suggest that pasireotide LAR may have a role in the treatment of acromegaly.[br][br]Sources of Research Support: Novartis Pharmaceuticals.[br][br]Disclosures: MDB: Consultant, Novartis Pharmaceuticals; Investigator, Novartis Pharmaceuticals. MS: Speaker, Novartis Pharmaceuticals. MG: Investigator, Novartis Pharmaceuticals; Speaker, Novartis Pharmaceuticals, Pfizer, Inc. MF: Researcher, Novartis Pharmaceuticals, Ipsen; Medical Advisory Board Member, Novartis Pharmaceuticals; Speaker, Ipsen, Chiasma. KHR: Employee, Novartis Pharmaceuticals. MR: Employee, Novartis Pharmaceuticals. YC: Employee, Novartis Pharmaceuticals. AC: Investigator, Novartis Pharmaceuticals. Nothing to Disclose: PF, FG, C-CS 2012-06-26T14:30:00 Theater C 2012-06-26T00:00:00 1899-12-30T14:30:00 1754 405 2837 OR49-3 OR22-02 Tuesday 2395 2012


2391 ENDO12L_OR49-4 ORAL SESSION: Novel Treatment Approaches for Pituitary Tumors (2:00 PM-3:30 PM) Octreolin[trade] Pharmacodynamics: An Oral Octreotide Formulation To Treat Acromegaly Suppresses Growth Hormone in Healthy Volunteers S L Teichman, S Tuvia, J Atsmon, S Katz, P Salama, D Pelled, I Landau, I Karmeli, W G Kramer, C J Strasburger, M Bidlingmaier, D L Kleinberg, S Melmed, R Mamluk Chiasma, Jerusalem, Israel; Tel Aviv Sourasky Medical Center, Affiliated to Sackler Faculty of Medicine, Tel Aviv, Israel; Kramer Consulting, LLC, North Potomac, MD; Campus Mitte, Charit[eacute] Universit[auml]tsmedizin Berlin, Berlin, Germany; Klinikum der Universit[auml]t, Munich, Germany; New York University, New York, NY; Cedars Sinai Medical Center, Los Angeles, CA [bold]Background:[/bold] An oral octreotide would enable a therapeutic alternative to the current injectable somatostatin analogs (SSTa or SRIFa) octreotide and lanreotide for chronic use in patients with acromegaly and other indications. A novel formulation of unmodified octreotide (Octreolin[trade]) was developed using the Transient Permeability Enhancer (TPE) technology. The TPE enables octreotide absorption by inducing a transient and reversible opening of the intestinal epithelial cell tight junctions. Seventy-four healthy volunteers have received one or more doses of Octreolin in several Phase 1 clinical studies. In pharmacokinetic (PK) studies, an oral octreotide dose of 20 mg resulted in circulating therapeutic levels that were comparable to a subcutaneous injection of 100 mcg.[br][bold]Objective:[/bold] To assess the pharmacodynamic (PD) effect of Octreolin on basal and stimulated growth hormone (GH) secretion.[br][bold]Methods:[/bold] Healthy men and women were enrolled in a crossover design with and without Octreolin dosing. Basal serum GH levels were measured in 16 subjects prior to (1 h) and for 2 h following 20 mg oral octreotide dosing. GH releasing-hormone (RH)/arginine was then administered as an iv bolus and GH levels measured for an additional 2 h.[br][bold]Results:[/bold] GH response to GHRH/arginine was blunted in all subjects receiving Octreolin with a trend towards increased suppression as drug exposure increased. Compared with the non-dosed period (mean [plusmn] standard error), Octreolin significantly suppressed mean basal GH secretion from 1.3[plusmn]0.4 to 0.5[plusmn]0.2 ng/mL (44%, p[lt]0.05) and mean GHRH-stimulated GH secretion from 56.1[plusmn]6.0 to 12.2[plusmn]3.6 ng/mL (80%, p[lt]0.001). The median duration of octreotide plasma levels [gt]0.5 ng/mL was 8 h. Octreolin was well tolerated with no significant adverse clinical events or laboratory abnormalities. Side effects of Octreolin were predominantly gastrointestinal and comparable in frequency and severity to those observed after injected octreotide, except for the absence of injection site complications.[br][bold]Conclusions:[/bold] Octreolin enables oral delivery of biologically active octreotide that achieves therapeutically relevant systemic blood levels. A single dose of Octreolin inhibits both basal and GHRH-induced GH secretion in healthy volunteers. Based on prior trials of injectable SRIFa and Octreolin clinical and PK data collected to date, a global Phase 3 efficacy and safety trial of Octreolin in patients with acromegaly is currently underway.[br][br]Sources of Research Support: Chiasma, Inc.[br][br]Disclosures: SLT: Employee, Chiasma. ST: Employee, Chiasma, Inc. JA: Principal Investigator, Chiasma. SK: Employee, Chiasma. PS: Employee, Chiasma. DP: Employee, Chiasma. IL: Employee, Chiasma. IK: Employee, Chiasma. WGK: Consultant, Chiasma. CJS: Medical Advisory Board Member, Chiasma. MB: Study Investigator, Chiasma. DLK: Medical Advisory Board Member, Chiasma. SM: Medical Advisory Board Member, Chiasma. RM: Employee, Chiasma. 2012-06-26T14:45:00 Theater C 2012-06-26T00:00:00 1899-12-30T14:45:00 281 405 2838 OR49-4 OR22-02 Tuesday 2396 2012


2392 ENDO12L_OR49-5 ORAL SESSION: Novel Treatment Approaches for Pituitary Tumors (2:00 PM-3:30 PM) Thrombospondin 1 (TSP-1) Analogs ABT-510 and ABT-898 Inhibit Prolactinoma Growth and Recover Pituitary Active Transforming Growth Factor b1 (TGF-b1) Maria V Recouvreux, Maria A Camilletti, Daniel B Rifkin, Damasia Becu-Villalobos, Graciela S Diaz-Torga IBYME-CONICET, Buenos Aires, Argentina; Langone Medical Center, New York University, New York, NY Prolactinomas are the most prevalent type of secreting pituitary tumors in humans, and generally respond well to a medical therapy with dopamine agonists. However, for patients exhibiting resistance to dopaminergic drugs, alternative treatments are desired. Antiangiogenic strategies might represent a potential therapy for these tumors. TSP-1 is a large multifunctional glycoprotein involved in multiple biological processes including angiogenesis, apoptosis, and activation of TGF-b1. Since tumors that over-express TSP-1 grow slower, have fewer metastases and have decreased angiogenesis, TSP-1 provides a novel target for cancer treatment. ABT-510 and ABT-898 (Abbott Laboratories) are TSP-1 synthetic analogues that mimic its antiangiogenic action.[br]The aim of the present study was to explore the potential therapeutic effect of ABT-510 and ABT-898, on experimental prolactinomas developed after 4 weeks of diethylstilbestrol (DES) treatment in female rats. Two weeks after DES pellet (20mg) implant, rats were injected three times a week during two weeks with vehicle, ABT-510 or ABT-898 (100mg/kg ip). Both TSP1 analogues were effective in decreasing pituitary tumor size (p=0.03) and PRL serum levels (P=0.0002), induced by DES. PCNA protein expression evaluated by western blot, was also reduced after treatment, indicating a decrease in tumor proliferation rate (P[lt]0.03). In order to evaluate the antiangiogenic effect of the drugs, we analyzed the vasculature by immunohistochemical staining of pituitary sections with the endothelial cell marker CD31. Total vascular area, as well as the number of CD31 positive vessels per area, were increased in DES pituitaries compared with controls (P[lt]0.03). Both ABT-510 and ABT-898 treatment significantly decreased microvessel density (P=0.01 and P=0.027). Finally, we found that treatment with the TSP-1 mimetic peptides were able to recover pituitary active TGF-b1 content in DES-treated rats, without effect on total TGF-b1 levels (measured by ELISA) or in its mRNA synthesis. Accordingly, we found that PSMAD2/3 expression was enhanced after treatment, suggesting a recovery of TGF-b1 activity in the pituitary.[br]Our results show that ABT-510 and ABT-898 decrease pituitary tumor angiogenesis, tumor size, proliferation rate, serum prolactin values, and restore pituitary TGF-b1 activity. These results, place these synthetic TSP-1 analogs as potential alternative or complementary treatments in dopamine agonist-resistant prolactinomas.[br][br]Sources of Research Support: CONICET: Consejo de Investigaciones Cientificas y Tecnicas. ANPCyT: Agencia Nacional de Promocion Cientifica y Tecnica.[br][br]Nothing to Disclose: MVR, MAC, DBR, DB-V, GSD-T 2012-06-26T15:00:00 Theater C 2012-06-26T00:00:00 1899-12-30T15:00:00 284 405 2839 OR49-5 OR22-02 Tuesday 2397 2012


2393 ENDO12L_OR49-6 ORAL SESSION: Novel Treatment Approaches for Pituitary Tumors (2:00 PM-3:30 PM) Acute Management of Pituitary Apoplexy: Experience in 80 Consecutive Cases Biju Jose, Lavanya Pelluri, Rosalind Mitchell, Neil Gittoes, John Ayuk University Hospital Birmingham, Birmingham, UK; University Hospital Birmingham, Birmingham, UK [bold]Introduction[/bold][br]Pituitary apoplexy is an uncommon medical emergency with incidence in pituitary adenomas of 2-7%. There are no evidence-based standards of optimum care for these patients. The key controversy in management relates to the role of acute neurosurgical intervention. The nature of clinical presentation precludes robust randomised controlled trials. Practical guidelines are derived from high-quality observational studies. On this background we report the management of pituitary apoplexy in a large cohort of patients treated at a single tertiary referral centre.[br][bold]Methods[/bold][br]A retrospective analysis evaluating clinical presentation, management and clinical outcomes in a cohort presenting with pituitary apoplexy during 1989-2010[br][bold]Results[/bold][br]80 patients (34 female) were included. Ten (12.5%) had previous diagnosis of pituitary tumour. The most common symptoms at presentation were headache (89%), visual disturbance (63%), cranial nerve involvement (43%) and diplopia (40%).[br]32 patients (40%) underwent surgery; 48 (60%) were managed conservatively. The indications for surgery were deteriorating vision/visual field (n=16), cranial nerve palsies (n=10) and tumour size (n=6).[br]All 32 patients in the surgical group had visual disturbance; 59% recovered fully. In the conservatively managed group, 33 had documented visual disturbance; 76% made a full recovery.[br]Pituitary function was assessed in all patients at presentation. Among the surgical group, 27 (84%) patients required hydrocortisone; 4/27 (15%) managed to withdraw treatment. 22 (69%) required thyroxine, none stopped. Eleven (34%) required sex steroids and none stopped. Three (9%) started growth hormone; one stopped.[br]Among the 48 conservatively managed patients, 38 (79%) started hydrocortisone; 5/38 (13%) stopped. 29 (60%) required thyroxine and none stopped. 22 (46%) started sex steroids, none stopped. Nine (19%) started growth hormone; none stopped.[br]At latest follow up, five patients were deceased from unrelated causes. Five patients in the surgical group required radiotherapy for enlarging tumours. In the conservative group, one patient each underwent surgery and radiotherapy, both for enlarging tumour.[br][bold]Conclusion[/bold][br]Our data represent the largest case series from a single centre. We propose that patients with acute apoplexy who have mild or stable symptoms/signs can be managed conservatively with careful monitoring; only rarely is there a need to change from conservative to surgical management in these patients.[br][br]Nothing to Disclose: BJ, LP, RM, NG, JA 2012-06-26T15:15:00 Theater C 2012-06-26T00:00:00 1899-12-30T15:15:00 1280 405 2840 OR49-6 OR22-02 Tuesday 2398 2012


2765 ENDO12L_M47 MEET-THE-PROFESSOR: CLINICAL - Raising HDL: When [amp] How? (Aim High) (2:45 PM - 3:30 PM) Raising HDL: When & How? (Aim High) Ira Jay Goldberg Columbia University, New York, NY Session supported by: Merck & Co., Inc. 2012-06-25T14:45:00 Room 342 ABDE 2012-06-25T00:00:00 1899-12-30T14:45:00 6290 329 2623 M47 L2 Monday 2012


3027 ENDO12L_M46 MEET-THE-PROFESSOR: CLINICAL - Minimally Invasive Surgery for Hyperparathyroidism (2:45 PM - 3:30 PM) Minimally Invasive Surgery for Hyperparathyroidism Robert Udelsman Yale University School of Medicine, New Haven, CT Nothing to Disclose: RU 2012-06-25T14:45:00 Room 361 ABDE 2012-06-25T00:00:00 1899-12-30T14:45:00 6208 328 2622 M46 B6 Monday 314 2012


3028 ENDO12L_M48 MEET-THE-PROFESSOR: CLINICAL - Update in FGF 23 (2:45 PM - 3:30 PM) Update in FGF 23 Michael John Econs Indiana University Medical Center, Indianapolis, IN Disclosures: MJE: Indiana University has a patent, but no clinical products at this time, Kirin Brewery; Consultant, Kirin Brewery. 2012-06-25T14:45:00 Room 352 DEF 2012-06-25T00:00:00 1899-12-30T14:45:00 6147 330 2624 M48 B10 Monday 315 2012


3029 ENDO12L_W2 WORKSHOP: TRANSLATIONAL - Health Disparities in Endocrine Disorders: Biological, Clinical [amp] Non-Clinical Factors: An Endocrine Society Scientific Statement (2:45 PM - 3:30 PM) Health Disparities in Endocrine Disorders: Biological, Clinical [amp] Non-Clinical Factors: An Endocrine Society Scientific Statement Sherita Hill Golden Johns Hopkins University Hospital, Baltimore, MD Nothing to Disclose: SHG 2012-06-25T14:45:00 Theater B 2012-06-25T00:00:00 1899-12-30T14:45:00 6215 331 2625 W2 W2 Monday 316 2012


3030 ENDO12L_PMW2 PRACTICE MANAGEMENT WORKSHOP: CLINICAL - The Endocrinologist Workforce: An Analysis of Supply [amp] Demand (2:45 PM - 3:30 PM) The Endocrinologist Workforce: An Analysis of Supply [amp] Demand Robert Alan Vigersky Walter Reed National Military Medical Center, Washington, DC Disclosure Incomplete: RAV 2012-06-25T14:45:00 Room 360 2012-06-25T00:00:00 1899-12-30T14:45:00 6207 332 2626 PMW2 PMW2 Monday 318 2012


3031 ENDO12L_PS6-1 PRESIDENTIAL SYMPOSIUM SESSION: CLINICAL - Reasons for Ethnic Disparities in Diabetes (3:45 PM - 5:15 PM) Reasons for Increased Diabetes Risk in Southeast Asians Abhimanyu Garg University Texas Southwest Medical Center, Dallas, TX Session supported by: Lilly USA, LLC[br][br]Nothing to Disclose: AG 2012-06-25T15:45:00 Theater B 2012-06-25T00:00:00 1899-12-30T15:45:00 6091 333 2627 PS6-1 T09-S01 Monday 320 2012


3032 ENDO12L_PS6-2 PRESIDENTIAL SYMPOSIUM SESSION: CLINICAL - Reasons for Ethnic Disparities in Diabetes (3:45 PM - 5:15 PM) Ethnic, Glucoregulation [amp] Cardiovascular Risk in African-Americans Kwame Osei Ohio State University Medical Center, Columbus, OH Session supported by: Lilly USA, LLC[br][br]Disclosure Incomplete: KO 2012-06-25T16:15:00 Theater B 2012-06-25T00:00:00 1899-12-30T16:15:00 6092 333 2628 PS6-2 T09-S01 Monday 321 2012


3033 ENDO12L_PS6-3 PRESIDENTIAL SYMPOSIUM SESSION: CLINICAL - Reasons for Ethnic Disparities in Diabetes (3:45 PM - 5:15 PM) Impact of Race/Ethnicity on the Effectiveness of Therapy in Diabetes Jaime A Davidson University of Texas Southwestern Medical Center, Dallas, TX Session supported by: Lilly USA, LLC[br][br]Disclosure Incomplete: JAD 2012-06-25T16:45:00 Theater B 2012-06-25T00:00:00 1899-12-30T16:45:00 6093 333 2629 PS6-3 T09-S01 Monday 322 2012


3034 ENDO12L_PS7-1 PRESIDENTIAL SYMPOSIUM SESSION: TRANSLATIONAL - Translational Insights into Rational Selection of Second Line Therapies for Progressive Endocrine Cancer (3:45 PM - 5:15 PM) Use of More Active Single Target Inhibitors When First Line Therapy Fails Sareh Parangi Massachusetts General Hospital/Harvard Medical School/Mt Auburn Hospital, Boston, MA Disclosure Incomplete: SP 2012-06-25T15:45:00 Room 332 2012-06-25T00:00:00 1899-12-30T15:45:00 6195 334 2630 PS7-1 SSS05 Monday 324 2012


3035 ENDO12L_PS7-2 PRESIDENTIAL SYMPOSIUM SESSION: TRANSLATIONAL - Translational Insights into Rational Selection of Second Line Therapies for Progressive Endocrine Cancer (3:45 PM - 5:15 PM) Discovery and Step-Wise Improvement of Cancer Therapeutics Using [italic]Drosophila[/italic] Chemical Genetics Tirtha K Das Mt Sinai School of Medicine, New York, NY Cancer therapeutics has two major challenges - to identify potent drug combinations as well as novel inhibitors with maximal therapeutic index. Rational drug combinations or novel polypharmacological inhibitors could target most pathways required for cancer progression and also control disease in patients with resistance to current first-line therapies. Traditional cell-line based screening methods are not always predictive of optimal therapeutic index. Using whole animal drug screening in a [italic]Drosophila[/italic] model of [italic]Multiple Endocrine Neoplasia Type II (MEN2)[/italic] we tested drug combinations and screened for novel inhibitors.[br][bold]Drug Combinations[/bold] [ndash] using a panel of 20 drugs, including drugs in clinical trials for [italic]MEN2[/italic], we tested a matrix that exceeded over 1000 drug-dose combinations in flies. We identified combinations that showed much higher suppression of [italic]MEN2[/italic] cancer cell line viability than individual drugs. Using [italic]Drosophila[/italic] genetics we identified a common pathway, which when inhibited, further improved the efficacy of some combinations. Including a third drug, that rationally inhibited this pathway, greatly improved the efficacy of the cocktail.[br][bold]Novel Inhibitors[/bold] [ndash] we screened a panel of novel polypharmacological compounds that inhibited RTK[apos]s as well as cellular kinases. One compound AD57 strongly suppressed oncogenic Ret induced phenotypes in flies. Chemical analogs AD36 and AD58 showed reduced efficacy and high toxicity respectively. Through [italic]Drosophila[/italic] genetics and comparison of in-vitro kinome inhibition profiles of the three compounds we identified Ret, Ras, Src, and PI3K pathways as candidates for inhibition. Surprisingly, inhibition of dTor led to paradoxical hyperactivation of the Ras/MAPK pathway. Chemical design based modification of AD57 led to development of AD80, which retained the desired targets of AD57 but eliminated dTor inhibition. AD80 showed significantly more efficacy than AD57, as well as currently approved therapies for [italic]MEN2[/italic], in both flies and mammalian models. Combining [italic]Drosophila[/italic] genetics, kinase chemistry, and kinome-wide profiling is a powerful approach for rationally identifying kinase inhibitors as well as drug combinations with maximal therapeutic index.[br][br]Nothing to Disclose: TKD 2012-06-25T16:15:00 Room 332 2012-06-25T00:00:00 1899-12-30T16:15:00 2503 334 2631 PS7-2 SSS05 Monday 325 2012


3036 ENDO12L_S43-1 SYMPOSIUM SESSION: TRANSLATIONAL - Adipose Tissue Inflammation [amp] Remodeling (3:45 PM - 5:15 PM) Adipose Remodeling and Obesity Philipp E Scherer University of Texas Southwestern Medical Center, Dallas, TX During the progression from the lean to the obese state, adipose tissue undergoes hyperplasia as well as hypertrophy in an attempt to cope with the increased demand for triglyceride storage. This requires a high degree of plasticity at both the cellular and at the tissue level. Even though adipose tissue as a whole seems to be a relatively static tissue containing many adipocytes that turn over slowly, these cells are embedded in an environment that can rapidly adapt to the needs of expanding and newly differentiating adipocytes. The extracellular matrix of adipose tissue faces unique challenges with respect to adjusting to the need for remodeling and expansion. In parallel, the vasculature has to adapt to altered requirements for nutrient and oxygen exchange. A decrease in the plasticity of these processes leads to metabolic dysfunction.[br]Excess levels of FFAs within adipose tissue have been shown to initiate a vicious cross-talk in between dysfunctional adipocytes and macrophages resulting in increased production of pro-inflammatory cytokines. This in turn leads to a state of chronic systemic inflammation, which further disrupts proper metabolic regulation. Recent studies pinpoint, however, that the interplay in between adipocytes and macrophages are much more dynamic than previously thought. Increased lipolysis during early phases of weight loss increases the infiltration of macrophages, a phenomenon that previously only has been associated with metabolic dysfunction in obesity. Furthermore, our recent observations show that fasting causes an increase in the circulating levels of the acute phase reactant serum Amyloid A (SAA) and induces a shift from M1 to M2 polarized macrophages in the adipose tissue. M1 macrophages are more pro-inflammatory and commonly associated with dysfunctional adipose tissue, while the M2-types are regarded as anti-inflammatory macrophages implicated in processes such as wound-healing. Obese adipose tissue commonly displays an increase in both M1- and M2-macrophages, with a propensity towards the M1 type. Interestingly, this fasting-induced shift from M1 to M2 does not occur under obese, insulin resistant conditions, indicating that the dynamic adipocyte-immune cell interplay is lost in metabolically compromised states.[br][br]Sources of Research Support: NIH R01-DK55758. NIH P01-DK088761.[br][br]Nothing to Disclose: PES 2012-06-25T15:45:00 Room 362 2012-06-25T00:00:00 1899-12-30T15:45:00 2418 335 2632 S43-1 T04-S04 Monday 327 2012


3037 ENDO12L_S43-2 SYMPOSIUM SESSION: TRANSLATIONAL - Adipose Tissue Inflammation [amp] Remodeling (3:45 PM - 5:15 PM) Immune Cell Function in Adipose Tissue Ajay Chawla University of California, San Francisco, San Francisco, CA Macrophages are immune sentinels that quiescently monitor their tissue milieu for early signs of infection, damage, or metabolic stress. In this role, the macrophage is responsible for sensing, integrating, and responding appropriately to a bewildering array of stimuli from its microenvironment. These pleiotropic responses are coordinated through distinct programs of macrophage activation, ranging from classical to alternative. We have previously shown that cooperative interactions between cytokine signaling pathways (IL-4 and IL-13) and metabolic regulators (PPARs) form the molecular switch that polarizes tissue macrophages to the alternative state. In metabolic disease, impairment in alternative activation of white adipose tissue macrophages (WAT) or Kupffer cells predisposes mice to diet-induced metabolic disease. Moreover, in WAT, eosinophils are competent for IL-4 production and necessary for alternative macrophage activation. Congruent with this observation, mice lacking eosinophils are predisposed to obesity and insulin resistance, whereas those with eosinophilia, either genetic or parasite induced, are protected from the deleterious effects of obesity. Together, these findings suggest that a hematopoietic circuit, consisting of eosinophils and alternatively activated macrophages, is a critical regulator of nutrient homeostasis and anabolic actions of insulin. Finally, new data will be presented to demonstrate the regulatory functions of alternatively activated macrophages in energy expenditure and thermogenesis.[br][br]Sources of Research Support: This work was supported by grants from: NIH (DK076760, HL076746, DK094641), Larry L. Hillblom Foundation Network Grant and an NIH Director[apos]s Pioneer Award (DP1OD006415) to A.C.[br][br]Nothing to Disclose: AC 2012-06-25T16:15:00 Room 362 2012-06-25T00:00:00 1899-12-30T16:15:00 2522 335 2633 S43-2 T04-S04 Monday 328 2012


3023 ENDO12L_M42 MEET-THE-PROFESSOR: CLINICAL - Assessing Readiness, Behavioral Modification [amp] Obesity (2:45 PM - 3:30 PM) Assessing Readiness, Behavioral Modification [amp] Obesity Daniel Holland Bessesen University of Colorado School of Medicine, Denver, CO Disclosure Incomplete: DHB 2012-06-25T14:45:00 Room 320 2012-06-25T00:00:00 1899-12-30T14:45:00 6164 324 2618 M42 OB1 Monday 310 2012


3024 ENDO12L_M43 MEET-THE-PROFESSOR: CLINICAL - Common Pitfalls in the Evaluation of Patients with Adrenal Incidentalomas (2:45 PM - 3:30 PM) Common Pitfalls in the Evaluation of Patients with Adrenal Incidentalomas Amir Hekmat Hamrahian Cleveland Clinic Foundation, Cleveland, OH The widespread use of abdominal imaging has resulted in an increased frequency of detected adrenal lesions. An adrenal incidentaloma is an adrenal mass, 1 cm or more in diameter, that is discovered serendipitously by radiological examination in the absence of symptoms or clinical findings suggestive of adrenal disease. When clinicians confront an adrenal incidentaloma, evaluation must address 2 issues: is the lesion functional or is it malignant and if so, is it primary or metastatic. In spite of the fact that the majority of adrenal incidentalomas are benign and non-functional, careful evaluation of all adrenal incidentalomas is warranted to ensure that tumors, such as primary adrenocortical carcinoma (PAC) and pheochromocytoma, that can be associated with significant morbidity and mortality, are not missed.[br]The adrenal computed tomographic (CT) is the cornerstone of imaging studies for adrenal tumors, providing information about size, density, presence of calcifications, areas of necrosis and extent of local invasion of the neoplasm. The three most important imaging criteria are: 1) size of the lesion, 2) the unenhanced CT attenuation value, and 3) the percentage washout. Several studies have shown that the non-contrast CT attenuation is superior to adrenal size in differentiating between benign and malignant adrenal tumors. Appropriate image interpretation can be a guide to a rational diagnostic workup and management.[br]There continues to be disagreement among experts on the choice and the extent of biochemical evaluation in patients with adrenal incidentalomas. All patients with an adrenal incidentaloma should be evaluated for autonomous cortisol secretion, termed subclinical Cushing[apos]s syndrome, (SCS), pheochromocytoma and, if hypertensive, primary aldosteronism. There is significant inconsistency among criteria used in the literature for the diagnosis of SCS. In spite of the lack of a single gold standard test to diagnose SCS, the 1-mg DST is currently considered the best diagnostic tool for evaluation of patients suspected to have SCS. Measurement of plasma metanephrines or 24 hour urine metanephrines are appropriate screening tests for pheochromocytoma. While measurement of plasma aldosterone to renin ratio (ARR) is considered as the best initial screening test for the evaluation of primary aldosteronism, there is a lack of consensus about what ARR cut-off should trigger further evaluation.[br][br]Nothing to Disclose: AHH 2012-06-25T14:45:00 Room 332 2012-06-25T00:00:00 1899-12-30T14:45:00 2459 325 2619 M43 A3 Monday 311 2012


3025 ENDO12L_M44 MEET-THE-PROFESSOR: CLINICAL - Management of Early Stage Cancer/Need for RAI (2:45 PM - 3:30 PM) Management of Early Stage Cancer/Need for RAI Bryan McIver Mayo Clinic, Rochester, MN Session supported by: Genzyme Corporation[br][br]Nothing to Disclose: BM 2012-06-25T14:45:00 Room 310 2012-06-25T00:00:00 1899-12-30T14:45:00 6160 326 2620 M44 TH3 Monday 312 2012


3026 ENDO12L_M45 MEET-THE-PROFESSOR: CLINICAL - Management of Gender Variant Youth (2:45 PM - 3:30 PM) Management of Gender Variant Youth Norman P Spack Children[apos]s Hospital Boston, Boston, MA Nothing to Disclose: NPS 2012-06-25T14:45:00 Room 370 2012-06-25T00:00:00 1899-12-30T14:45:00 6184 327 2621 M45 PED4 Monday 313 2012


3072 ENDO12L_M50 MEET-THE-PROFESSOR: CLINICAL - Biochemical Markers of Bone Turnover in Osteoporosis (8:30 AM - 9:15 AM) Biochemical Markers of Bone Turnover in Osteoporosis Roberto Civitelli Washington University -St Louis, Saint Louis, MO Disclosures: RC: Stockholder, Eli Lilly & Company, Amgen, Merck & Co.; Principal Investigator, Pfizer, Inc. 2012-06-26T08:30:00 Room 370 2012-06-26T00:00:00 1899-12-30T08:30:00 6174 361 2680 M50 B9 Tuesday 370 2012


3073 ENDO12L_M51 MEET-THE-PROFESSOR: CLINICAL - Congenital Adrenal Hyperplasia from Adolescent to Adult (8:30 AM - 9:15 AM) Congenital Adrenal Hyperplasia from Adolescent to Adult Richard Joseph Auchus University of Michigan, Ann Arbor, MI Disclosures: RJA: Investigator, Jansen Pharmaceuticals. 2012-06-26T08:30:00 Room 372 2012-06-26T00:00:00 1899-12-30T08:30:00 6136 358 2676 M51 A1 Tuesday 371 2012


3074 ENDO12L_M52 MEET-THE-PROFESSOR: CLINICAL - Diagnosis [amp] Treatment of Type 2 Diabetes in Youth (8:30 AM - 9:15 AM) Diagnosis [amp] Treatment of Type 2 Diabetes in Youth Francine Ratner Kaufman University Childrens Medical Group, Los Angeles, CA Session supported by: Lilly USA, LLC[br][br]Disclosures: FRK: Chief Scientific Officer, Medtronic Minimed. 2012-06-26T08:30:00 Room 332 2012-06-26T00:00:00 1899-12-30T08:30:00 6163 362 2681 M52 D8 Tuesday 372 2012


3075 ENDO12L_M53 MEET-THE-PROFESSOR: CLINICAL - Pediatric Type 1 Diabetes (8:30 AM - 9:15 AM) Pediatric Type 1 Diabetes Morey W Haymond Children[apos]s Nutrition Research Center, Houston, TX Session supported by: Lilly USA, LLC[br][br]Disclosure Incomplete: MWH 2012-06-26T08:30:00 Room 320 2012-06-26T00:00:00 1899-12-30T08:30:00 6244 363 2682 M53 D2 Tuesday 373 2012


3076 ENDO12L_M54 MEET-THE-PROFESSOR: CLINICAL - Tumor-Induced Osteomalacia (8:30 AM - 9:15 AM) Tumor-Induced Osteomalacia Suzanne Marie Jan De Beur Johns Hopkins University School of Medicine, Baltimore, MD Disclosure Incomplete: SMJDB 2012-06-26T08:30:00 Room 361 ABDE 2012-06-26T00:00:00 1899-12-30T08:30:00 6140 364 2683 M54 B1 Tuesday 374 2012


3077 ENDO12L_S50-1 SYMPOSIUM SESSION: TRANSLATIONAL - Cross-Talk of Androgen Receptor with Other Signaling Pathways (9:30 AM - 11:00 AM) Androgen Receptor-Cyclin Interactions Karen E Knudsen Thomas Jefferson University, Philadelphia, PA Nothing to Disclose: KEK 2012-06-26T09:30:00 Room 361 2012-06-26T00:00:00 1899-12-30T09:30:00 6081 365 2684 S50-1 T08-S01 Tuesday 376 2012


3078 ENDO12L_S50-2 SYMPOSIUM SESSION: TRANSLATIONAL - Cross-Talk of Androgen Receptor with Other Signaling Pathways (9:30 AM - 11:00 AM) Targeting the Regulation of Androgen Receptor by the 52 kDa FK506-Binding Protein (FKBP52) for the Treatment of Prostate Cancer Marc B Cox University of Texas at El Paso, El Paso, TX Drugs that target novel surfaces on the androgen receptor and/or novel AR regulatory mechanisms are promising alternatives for the treatment of hormone refractory prostate cancer (HRPC). The 52 kDa FK506-Binding Protein (FKBP52) has been shown to be an important positive regulator of AR in cellular and whole animal models and represents an attractive target for the treatment of prostate cancer. We have identified a surface region on the androgen receptor (AR) hormone binding domain that, when mutated, displays a greater dependence on FKBP52 for normal function. Interestingly, the BF3 surface is also hypothesized to participate in the interaction of AR with the co-activator [beta]-catenin, and co-expression of FKBP52 and [beta]-catenin act in synergy to up-regulate both hormone-dependent and hormone-independent AR function. In addition, we have developed a series of small molecules that effectively inhibit the FKBP52 regulation of AR function and the synergistic up-regulation of AR function by FKBP52 and [beta]-catenin. Surface plasmon resonance studies have confirmed that these inhibitors disrupt the regulation of AR by FKBP52 and [beta]-catenin through interaction with the AR hormone binding domain. We have demonstrated that these novel compounds do not compete with hormone for binding the hormone binding pocket, nor do they compete with coactivator peptide for binding AF2. In addition, we have shown that these compounds inhibit AR function by preventing hormone-dependent dissociation of the Hsp90-FKBP52-AR complex which results in less hormone-bound receptor in the nucleus. Assays in early and late stage prostate cancer cells have demonstrated that these novel compounds inhibit both prostate specific antigen expression and androgen-dependent proliferation. In summary, we have identified a putative FKBP52 interaction surface on the AR hormone binding domain and identified a series of small molecules that inhibit AR function by targeting that surface. This class of compounds, now termed NR alternate site modulators (NRAMs), would be useful in hormone-resistant prostate cancer, where coregulator overexpression and other mechanisms result in receptor activation in the absence of exogenous ligands. In addition, we have identified a novel functional interaction between FKBP52 and [beta]-catenin, which has clear implications in prostate cancer progression to the hormone refractory state.[br][br]Sources of Research Support: NIH Grant 5G12RR008124 awarded to the Border Biomedical Research Center (BBRC)/University of Texas at El Paso, ARRA funds through NIH Grant SC1GM084863, Cancer Prevention and Research Institute of Texas (CPRIT) Grant RP110444-P2, and RISE (R25GM069621) and NSF LSAMP (HRD-0832951) fellowships.[br][br]Nothing to Disclose: MBC 2012-06-26T10:00:00 Room 361 2012-06-26T00:00:00 1899-12-30T10:00:00 126 365 2685 S50-2 T08-S01 Tuesday 377 2012


3079 ENDO12L_S50-3 SYMPOSIUM SESSION: TRANSLATIONAL - Cross-Talk of Androgen Receptor with Other Signaling Pathways (9:30 AM - 11:00 AM) Interleukin Signaling in Androgen Action Zoran Culig Innsbruck Medical University, Innsbruck, Austria Interleukin-6 (IL-6) is implicated in regulation of proliferation, apoptosis, and angiogenesis in human prostate cancer. It was demonstrated that IL-6 activates androgen receptor in several human prostate cancer cell lines and the coactivators p300 and SRC-1 are required for this activation. AR activation by IL-6 may result in either stimulation or inhibition of proliferation, as evidenced in LNCaP and MDA PCa 2b cells, respectively. The therapeutic inhibition of IL-6 in vitro and in vivo could be achieved by an anti-IL-6 antibody siltuximab. This antibody delayed prostate cancer progression in a xenograft model. In the phase I clinical study, siltuximab has inhibited intracrine androgen production which clearly contributes to prostate cancer progression.[br]The effect of IL-6 in target tissues depends on the presence of intracellular negative regulators. We could demonstrate that protein inhibitor of activated STAT1 (PIAS1), which is a known coactivator of the androgen receptor, is expressed in the majority of cell lines in the prostate. It is also expressed at a higher level in malignant tissue compared to the benign compartment. PIAS1 down-regulation yielded inhibition of cell growth and its overexpression promoted growth of several prostate cancer cell lines. PIAS1 control of cell cycle progression is achieved through p21 down-regulation. Other molecules which may influence the outcome of IL-6 action are suppressors of cytokine signaling -3 and -1 (SOCS). SOCS-3 prevents apoptosis through modulation of both extrinsic and intrinsic pathways. However, it could also inhibit the effects of basic fibroblast growth factor which acts through the mitogen-activated protein kinase pathway. SOCS-3 could be induced by androgenic hormones. Interestingly, in conditions in which SOCS-3 is up-regulated androgenic effect on proliferation and secretion is diminished. SOCS-3 and -1 are expressed in specimens obtained from patiens with prostate cancer.[br]In summary, androgens and IL-6 interact at different levels in prostate cancer. A better understanding of implications of AR activation by IL-6 is important for the improvement of anti-IL-6 therapies in prostate cancer.[br][br]Sources of Research Support: Austrian Research Fund FWF, Centocor.[br][br]Disclosures: ZC: Principal Investigator, Johnson & Johnson. 2012-06-26T10:30:00 Room 361 2012-06-26T00:00:00 1899-12-30T10:30:00 2453 365 2686 S50-3 T08-S01 Tuesday 378 2012


3080 ENDO12L_S51-1 SYMPOSIUM SESSION: CLINICAL - CVD Risk in Diabetes [ndash] Is Glucose Important? (9:30 AM - 11:00 AM) Is Hyperglycemia a CVD Risk Factor? Hertzel C Gerstein McMaster University, Hamilton, Canada A diagnosis of diabetes mellitus is made when fasting or post-load glucose levels are consistently [underline][gt][/underline] 7.0 and 11.0 mmol/L respectively, and/or when an A1C level is consistently [underline][gt][/underline] 6.5%. These thresholds were based on prospective epidemiologic studies showing that levels above these thresholds are a strong risk factor for retinopathy. They were not chosen on the basis of risk for other serious outcomes despite the fact that prospective studies have consistently shown that diabetes is an independent risk factor for such outcomes including cardiovascular disease, cirrhosis, death, and cancer. Indeed, during the last 15 years, multiple epidemiologic analyses have consistently shown that the diagnostic glycemic thresholds for diabetes are not relevant for most of the health consequences of diabetes and have suggested that there is a graded relationship between glucose or A1C levels and serious health outcomes that extends from the upper limit of normal (e.g. a fasting plasma glucose of [sim] 5.6 mmol/l or 100 mg/dl) right into the diabetes range (i.e. dysglycemia). For example, in a recent epidemiologic analysis of population data from people with no history of diabetes or CVD (1) a 1% higher A1C predicted a 50% higher risk of coronary heart disease and a 50% higher risk of stroke.[br]Whether this relationship is caused of promoted by the dysglycemic state per se, by the associated relative insulin insufficiency that must be present to allow the glucose level to rise, by other associated biologic abnormalities, or by some factor that is confounded with both dysglycemia and serious health outcomes remains unknown. Also unknown is whether targeting, reversing or preventing early dysglycemia with lifestyle, insulin, incretins or oral agents can reduce the risk of many of these serious health outcomes. Such information can only be reliably obtained from large randomized trials which to date have yielded mixed results suggesting a possible modest effect on cardiovascular outcomes.[br][br]Session supported by: Merck & Co., Inc.[br][br]1. Selvin et al., NEJM 2010;362:800.[br][br]Disclosures: HCG: Speaker, Sanofi-Aventis; Ad Hoc Consultant, Sanofi-Aventis, Novo Nordisk, Merck & Co., Jansen Pharmaceuticals, Bristol-Myers Squibb; Research Funding, Sanofi-Aventis, Lilly USA, LLC; Consultant, Roche Pharmaceuticals, Abbott Laboratories; Scientific Board Member, Bayer, Inc. 2012-06-26T09:30:00 Theater C 2012-06-26T00:00:00 1899-12-30T09:30:00 2397 366 2687 S51-1 T09-S03 Tuesday 380 2012


3081 ENDO12L_S51-2 SYMPOSIUM SESSION: CLINICAL - CVD Risk in Diabetes [ndash] Is Glucose Important? (9:30 AM - 11:00 AM) Glycemic Variability [amp] CVD Risk Paresh Dandona Fillmore Hospital/State University of New York, Buffalo, NY Session supported by: Merck & Co., Inc.[br][br]Disclosure Incomplete: PD 2012-06-26T10:00:00 Theater C 2012-06-26T00:00:00 1899-12-30T10:00:00 6098 366 2688 S51-2 T09-S03 Tuesday 381 2012


3082 ENDO12L_S51-3 SYMPOSIUM SESSION: CLINICAL - CVD Risk in Diabetes [ndash] Is Glucose Important? (9:30 AM - 11:00 AM) Risk Factor Modification beyond Glucose Jane E Reusch Denver VAMC, Denver, CO Cardiovascular disease including ischemic heart disease, stroke, peripheral vascular disease and congestive heart failure contribute significantly to the excess morbidity and mortality in diabetes and shorten life expectancy. Much research has been focused upon glucose lowering to decrease cardiovascular mortality in diabetes. In this presentation, we will discuss other co-morbidities including sedentary lifestyle, hypertension, hyperlipidemia, and increased thrombogenicity as modifiable risk factors in people with diabetes.[br]Sedentary lifestyle has recently been characterized by the CDC as a leading cause of morbidity and mortality. Our research group has defined a defect in functional exercise capacity both submaximal in maximal in people with uncomplicated diabetes. Decreased physical fitness predicts premature mortality and may contribute to short life expectancy in people with diabetes.[br]Hypertension is a common feature of diabetes. Prospective interventional studies have demonstrated unequivocal benefit for antihypertensive therapy in people with diabetes. In addition, blood pressure lowering has a positive impact on progression of nephropathy. Studies defending aggressive blood pressure management in diabetes will be reviewed.[br]Hypercholesterolemia is a modifiable risk factor for the prevention of excess cardiovascular disease in diabetes. The lipid profile in most people with type 2 diabetes mellitus includes a moderate elevation in LDL cholesterol, with a more robust increase in triglycerides and low HDL cholesterol. Prospective randomized studies using HMG coA reductase inhibitors to lower cholesterol have demonstrated unequivocal efficacy. These studies will be reviewed as will studies with additional agents for cholesterol lowering.[br]Aspirin therapy is of high theoretical benefit people with diabetes. Despite that, large clinical trials and meta-analyses have demonstrated modest and often not statistically significant improvement in cardiovascular and all cause mortality with aspirin therapy. Recently, the guidelines for aspirin therapy in with diabetes will be reviewed.[br]Strategic cardiovascular risk factor reduction in should be implemented in all people with diabetes. As such people with diabetes should be approached with a multi-factorial approach. Large clinical trials such as STENO2, Look-AHEAD, ADDITION, ATTEMPT and others have and will demonstrate the need for a multifaceted intervention to decrease cardiovascular risk in diabetes.[br][br]Session supported by: Merck & Co., Inc.[br][br]Disclosures: JER: Principal Investigator, Amylin Pharmaceuticals, Bristol-Myers Squibb; Consultant, GlaxoSmithKline. 2012-06-26T10:30:00 Theater C 2012-06-26T00:00:00 1899-12-30T10:30:00 2468 366 2689 S51-3 T09-S03 Tuesday 382 2012


3083 ENDO12L_S52-1 SYMPOSIUM SESSION: TRANSLATIONAL - Gastrointestinal Neuroendocrine Tumors (9:30 AM - 11:00 AM) Update on the Genomics of Pancreatic Neuroendocrine Tumors Nickolas Papadopoulos Johns Hopkins, Baltimore Disclosures: NP: Founder, Inostics, PGDx, Inc. 2012-06-26T09:30:00 Room 320 2012-06-26T00:00:00 1899-12-30T09:30:00 6012 367 2690 S52-1 T02-S01 Tuesday 384 2012


3084 ENDO12L_S52-2 SYMPOSIUM SESSION: TRANSLATIONAL - Gastrointestinal Neuroendocrine Tumors (9:30 AM - 11:00 AM) Role of Aggressive Surgery in GI Neuroendocrine Tumor Steven K Libutti Albert Einstein College of Medicine, Bronx, NY Gastrointestinal and pancreatic neuroendocrine tumors are increasing in incidence (1). Surgical resection is the mainstay of therapy for primary GI and pancreatic neuroendocrine tumors. For metastatic disease, surgery can also be very effective for those patients with disease that is amenable to an R0 or R1 resection. Aggressive metastasectomy has been shown to impact recurrence free and perhaps overall survival in selected patients (2). For those patients for whom an R0 or R1 resection is not possible, yet have disease isolated to the liver, chemoperfusion therapies may offer an alternative to systemic therapy, TACE and radiotherapies (3,4). Isolated hepatic perfusion and percutaneous hepatic perfusion with chemosaturation both offer the potential to achieve high response rates and meaningful progression free survival for patients with unresectable NETs metastatic to the liver (3). These techniques are also very effective in controlling the symptoms of functional pancreatic NETs and carcinoid syndrome (3). With the recent development of targeted therapies such as everolimus and sunitinib (5), the potential to combine aggressive surgical resection with post-operative adjuvant therapies is now being explored. This presentation will discuss the indications and role for aggressive surgical resection for the management of GI and pancreatic NETs and the role of newer regional and targeted therapies in the treatment of these diseases.[br][br](1) Yao JC, Hassan M, Phan A, Dagohoy C, Leary C, Mares JE, Abdalla EK, Fleming JB, Vauthey JN, Rashid A, Evans DB. One hundred years after [quot]carcinoid[quot]: epidemiology of and prognostic factors for neuroendocrine tumors in 35,825 cases in the United States. J Clin Oncol. 2008 Jun 20;26(18):3063-72. Review. PubMed PMID: 18565894. (2) Mayo SC, de Jong MC, Pulitano C, Clary BM, Reddy SK, Gamblin TC, Celinksi SA,. Kooby DA, Staley CA, Stokes JB, Chu CK, Ferrero A, Schulick RD, Choti MA, Mentha. G, Strub J, Bauer TW, Adams RB, Aldrighetti L, Capussotti L, Pawlik TM. Surgical. management of hepatic neuroendocrine tumor metastasis: results from an. international multi-institutional analysis. Ann Surg Oncol. 2010. Dec;17(12):3129-36. Epub 2010 Jun 29. PubMed PMID: 20585879. (3) Grover AC, Libutti SK, Pingpank JF, Helsabeck C, Beresnev T, Alexander HR Jr. Isolated hepatic perfusion for the treatment of patients with advanced liver. metastases from pancreatic and gastrointestinal neuroendocrine neoplasms. Surgery. 2004 Dec;136(6):1176-82. PubMed PMID: 15657573. (4) Lewis MA, Hubbard J. Multimodal liver-directed management of neuroendocrine. hepatic metastases. Int J Hepatol. 2011;2011:452343. Epub 2011 Oct 29. PubMed. PMID: 22121491; PubMed Central PMCID: PMC3205732. (5) Dong M, Phan AT, Yao JC. New Strategies for Advanced Neuroendocrine Tumors in. the Era of Targeted Therapy. Clin Cancer Res. 2012 Feb 15. [Epub ahead of print]. PubMed PMID: 22338018.[br][br]Nothing to Disclose: SKL 2012-06-26T10:00:00 Room 320 2012-06-26T00:00:00 1899-12-30T10:00:00 2421 367 2691 S52-2 T02-S01 Tuesday 385 2012


3085 ENDO12L_S52-3 SYMPOSIUM SESSION: TRANSLATIONAL - Gastrointestinal Neuroendocrine Tumors (9:30 AM - 11:00 AM) New Targeted Therapies for Neuroendocrine Tumor Marianne E Pavel Charit[eacute], Berlin, Germany Disclosures: MEP: Consultant, Ipsen, Novartis Pharmaceuticals, Pfizer, Inc.; Speaker, Ipsen, Novartis Pharmaceuticals, Pfizer, Inc. 2012-06-26T10:30:00 Room 320 2012-06-26T00:00:00 1899-12-30T10:30:00 6013 367 2692 S52-3 T02-S01 Tuesday 386 2012


3086 ENDO12L_S53-1 SYMPOSIUM SESSION: CLINICAL - Hormonal Contraceptives [amp] Replacement Therapy in Adolescents (9:30 AM - 11:00 AM) Hormonal Contraceptives and Thrombotic Risk Catherine Mason Gordon Children[apos]s Hospital, Boston, MA Heightened publicity about the association between hormonal contraception and thrombotic risk, and the publication of new guidelines by the World Health Organization in 2009 and the Centers for Disease Control and Prevention in 2010 addressing this issue have led to multidisciplinary discussions regarding the appropriate care for adolescent patients. This lecture will outline an approach to the adolescent girl or young woman who presents for contraceptive counseling or advice on estrogen replacement therapy, including medically complex patients. The relative risk of thrombosis on combined oral contraception is three- to fivefold, whereas the absolute risk for a healthy adolescent on this therapy is only 0.05% per year. This thrombotic risk is affected by estrogen dose, type of progestin, mechanism of delivery, and length of therapy. As will be reviewed, oral progestin-only contraceptives and transdermal estradiol used for hormone replacement carry minimal or no thrombotic risk. However, transdermal, vaginal, or intrauterine contraceptives and injectable progestins merit further study. A personal history of thrombosis, persistent or inherited thrombophilia, and numerous lifestyle choices also influence thrombotic risk. This lecture will review relative-risk data and discuss the application of absolute risk to individual patient counseling. An approach to challenging patients with a history of thrombosis, known thrombophilia, or family history of thrombosis or thrombophilia will be discussed. As will be reviewed, knowledge of the guidelines and individualized management plans can be extremely useful for informing practice decisions around hormonal and nonhormonal options.[br][br]Disclosures: CMG: Co-Director Fellowship Training Program, Pfizer, Inc., Merck & Co. 2012-06-26T09:30:00 Room 332 2012-06-26T00:00:00 1899-12-30T09:30:00 2498 368 2693 S53-1 T07-S02 Tuesday 388 2012


3087 ENDO12L_S53-2 SYMPOSIUM SESSION: CLINICAL - Hormonal Contraceptives [amp] Replacement Therapy in Adolescents (9:30 AM - 11:00 AM) Is There a Hormonal Regimen of Choice for PCOS in Adolescence? Hala Mounir Tfayli American University of Beirut, Beirut, Lebanon Polycystic Ovary Syndrome (PCOS), a heterogeneous disorder characterized by androgen excess, irregular menses and/or cystic ovarian morphology, has peri-pubertal onset. In addition to androgen excess and reproductive disturbances, some adolescents with PCOS are at higher risk of metabolic derangements with longterm health sequelae. Obesity, impaired glucose tolerance, diabetes and the metabolic syndrome are highly prevalent among these youth. Insulin resistance and the consequent hyperinsulinemia are believed to play a pivotal role in the development of the PCOS associated metabolic disturbances and in promoting an ongoing state of androgen excess. Given that PCOS is not only a reproductive but also a metabolic disorder starting early in adolescence necessitates that therapeutic options target the hormonal as well as the metabolic disturbances. The conventional treatment of PCOS has been oral contraceptives (OCPs) and anti-androgenic agents. While OCPs are effective in attenuating hyperandrogenemia and the associated clinical manifestations, they have been reported to worsen lipid profile, increase inflammatory markers and decrease insulin sensitivity in some studies, but not in others. Insulin sensitizers such as metformin have emerged as potential alternatives with promising results on both the reproductive and the metabolic aspects. The thiazolidinediones (TZDs) have also been studied but to a more limited extent among adolescents with PCOS. While insulin sensitizers seem to have a more favorable outcome on the metabolic profile, their effect in attenuating hyperandrogenemia, especially hirsutism, may be less marked. Combination treatments, metformin with OCPs or with anti-androgens have also been studied with some promising results. The choice of treatment modality in this age group is limited by the paucity of randomized controlled trials, and by the lack of data on the longterm effects of therapeutic interventions initiated in adolescent years on the reproductive and cardiometabolic profile in adulthood. Moreover clinical decision making has to address the individual priorities of each patient.Considerations include treatment for hirsutism and acne, reproduction or contraception, weight gain and metabolic disturbances, within the context of genetic and familial risk for cardio-metabolic disorders. In this session we will review the available data in the adolescent age group regarding different treatment modalities.[br][br](1)Azziz R, Carmina E, Dewailly D, Diamanti-Kandarakis E, Escobar-Morreale HF, Futterweit W, Janssen OE, Legro RS, Norman RJ, Taylor AE, Witchel SF; Androgen Excess Society. J Clin Endocrinol Metab. 2006; 91(11):4237-45. (2) Tfayli H, Arslanian S, Ann N Y Acad Sci. 2008;1135:85-94. (3) Geller D, Pacaud D, Gordon C, Misra M, Int J Pediatr Endocrinol. 2011; 26;2011:9. (4) Arslanian SA, Lewy V, Danadian K, Saad R. J Clin Endocrinol Metab. 2002 Apr;87(4):1555-9. (5) Tfayli H, Ulnach JW, Lee S, Sutton-Tyrrell K, Arslanian S. J Clin Endocrinol Metab. 2011;96(5):1311-9. (6) Ibanez L, Diaz M, Sebastiani G, Sanchez-Infantes D, Salvador C, Lopez-Bermejo A, de Zegher F. J Clin Endocrinol Metab. 2011;96(11):3361-6.[br][br]Nothing to Disclose: HMT 2012-06-26T10:00:00 Room 332 2012-06-26T00:00:00 1899-12-30T10:00:00 2511 368 2694 S53-2 T07-S02 Tuesday 389 2012


3164 ENDO12L_M20A MEET-THE-PROFESSOR: CLINICAL - Pituitary Incidentaloma (1:00 PM - 1:45 PM) Pituitary Incidentaloma Mark E Molitch Northwestern University Feinberg Medical School, Chicago, IL Disclosure Incomplete: MEM 2012-06-24T13:00:00 Room 342 ABDE 2012-06-24T00:00:00 1899-12-30T13:00:00 6211 167 1046 M20 PIT1A Sunday 127 2012


3165 ENDO12L_M21A MEET-THE-PROFESSOR: CLINICAL - Molecular Markers for the Diagnosis of Thyroid Cancer by Fine Needle Aspiration (1:00 PM - 1:45 PM) Role of Molecular Markers in Cancer Jennifer Anne Sipos Ohio State University, Columbus, OH Session supported by: Genzyme Corporation[br][br]Disclosure Incomplete: JAS 2012-06-24T13:00:00 Room 320 2012-06-24T00:00:00 1899-12-30T13:00:00 6159 168 1047 M21 TH2A Sunday 128 2012


3166 ENDO12L_M22A MEET-THE-PROFESSOR: CLINICAL - The Very Insulin-Resistant Diabetes Patient: Work Up [amp] Treatment (1:00 PM - 1:45 PM) The Very Insulin-Resistant Diabetes Patient: Work Up [amp] Treatment Lisa R Tannock University of Kentucky, Lexington, KY Session supported by: Lilly USA, LLC[br][br]Nothing to Disclose: LRT 2012-06-24T13:00:00 Room 372 2012-06-24T00:00:00 1899-12-30T13:00:00 6148 169 1048 M22 D10A Sunday 129 2012


3167 ENDO12L_M23A MEET-THE-PROFESSOR: CLINICAL - There[apos]s an App for That: Mobile Health in the Management of Diabetes (1:00 PM - 1:45 PM) There[apos]s an App for That: Mobile Health in the Management of Diabetes [amp] Endocrine Conditions Ronald Tamler Mt Sinai Medical Center, New York, NY Disclosures: RT: Consultant, Sanofi-Aventis. 2012-06-24T13:00:00 Room 332 2012-06-24T00:00:00 1899-12-30T13:00:00 6192 170 1049 M23 D5A Sunday 130 2012


3168 ENDO12L_NS5-1A ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (2:00 PM - 3:00 PM) Carbohydrate Counting Lynn C Coppolo Bassett Healthcare, Cherry Valley, NY Disclosure Incomplete: LCC 2012-06-24T14:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T14:00:00 6239 206 1757 NS5-1 ENS5A Sunday 185 2012


3169 ENDO12L_NS5-2A ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (2:00 PM - 3:00 PM) Exercise Beth Lucasey TVAS BioMedical, Inc, Lenexa, KS Disclosure Incomplete: BL 2012-06-24T14:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T14:00:00 6234 206 1758 NS5-2 ENS5A Sunday 186 2012


3170 ENDO12L_NS5-3A ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (2:00 PM - 3:00 PM) Endocrine Stimulation Testing Roberta M Hower Lehigh Valley Hospital, Allentown, PA Disclosure Incomplete: RMH 2012-06-24T14:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T14:00:00 6235 206 1759 NS5-3 ENS5A Sunday 187 2012


3171 ENDO12L_NS5-4A ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (2:00 PM - 3:00 PM) ENDO 101/Pituitary Michelle H Gurel Massachusetts General Hospital, Boston, MA Disclosure Incomplete: MHG 2012-06-24T14:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T14:00:00 6236 206 1760 NS5-4 ENS5A Sunday 188 2012


3172 ENDO12L_NS5-5A ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (2:00 PM - 3:00 PM) Research Margaret E Eckert-Norton SUNY Downstate, Brooklyn, NY Disclosure Incomplete: MEE-N 2012-06-24T14:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T14:00:00 6237 206 1761 NS5-5 ENS5A Sunday 189 2012


3173 ENDO12L_NS5-6A ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (2:00 PM - 3:00 PM) Research Patricia S Via McGuire Veterans Affairs Medical Center, Richmond, VA Disclosure Incomplete: PSV 2012-06-24T14:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T14:00:00 6238 206 1762 NS5-6 ENS5A Sunday 190 2012


3174 ENDO12L_NS5-7A ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (2:00 PM - 3:00 PM) Insulin Management Elaine McLeod Tennessee Valley Healthcare System, Nashville, TN 2012-06-24T14:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T14:00:00 6291 206 1763 NS5-7 ENS5A Sunday 190 2012


3175 ENDO12L_CMF6-1A CASE MANAGEMENT FORUM: CLINICAL - Inpatient Management of Diabetes (5:30 PM - 6:15 PM) Inpatient Management of Diabetes Andrew J Ahmann Oregon Health Sciences University, Portland, OR Disclosure Incomplete: AJA 2012-06-24T17:30:00 Theater A 2012-06-24T00:00:00 1899-12-30T17:30:00 6209 232 1813 CMF6-1 CMF-D1A Sunday 192 2012


3176 ENDO12L_CMF6-2A CASE MANAGEMENT FORUM: CLINICAL - Inpatient Management of Diabetes (5:30 PM - 6:15 PM) Inpatient Management of Diabetes Guillermo Enrique Umpierrez Emory University School of Medicine, Atlanta, GA Nothing to Disclose: GEU 2012-06-24T17:30:00 Theater A 2012-06-24T00:00:00 1899-12-30T17:30:00 6187 232 1814 CMF6-2 CMF-D1A Sunday 193 2012


3177 ENDO12L_CMF7-1A CASE MANAGEMENT FORUM: CLINICAL - Reconciling Diabetes Treatment Algorithms Based on Evidence (5:30 PM - 6:15 PM) Reconciling Diabetes Treatment Algorithms Based on Evidence Lawrence Blonde Ochsner Medical Center, New Orleans, LA Session supported by: Lilly USA, LLC[br][br]Disclosures: LB: Principal Investigator, Eli Lilly & Company Novo Nordisk, Sanofi-Aventis; Speaker, Novo Nordisk, Boehringer Ingelheim, Merck & Co., Johnson & Johnson Diabetes Institute, L.L.C., SANTARUS; Scientific Board Member, Novo Nordisk, GlaxoSmithKline, Merck & Co., Sanofi-Aventis; Consultant, Amylin Pharmaceuticals, SANTARUS; Speaker Bureau Member, Amylin Pharmaceuticals; Owner, Pfizer, Inc., Amylin Pharmaceuticals. 2012-06-24T17:30:00 Room 332 2012-06-24T00:00:00 1899-12-30T17:30:00 6189 233 1815 CMF7-1 CMF-D2A Sunday 194 2012


3178 ENDO12L_CMF7-2A CASE MANAGEMENT FORUM: CLINICAL - Reconciling Diabetes Treatment Algorithms Based on Evidence (5:30 PM - 6:15 PM) Reconciling Diabetes Treatment Algorithms Based on Evidence Jaime A Davidson University of Texas Southwestern Medical Center, Dallas, TX Session supported by: Lilly USA, LLC[br][br]Disclosure Incomplete: JAD 2012-06-24T17:30:00 Room 332 2012-06-24T00:00:00 1899-12-30T17:30:00 6247 233 1816 CMF7-2 CMF-D2A Sunday 195 2012


3179 ENDO12L_M24A MEET-THE-PROFESSOR: CLINICAL - Adrenal Insufficiency (5:30 PM - 6:15 PM) Adrenal Insufficiency Baha M Arafah Case Western Reserve University, Cleveland, OH Nothing to Disclose: BMA 2012-06-24T17:30:00 Room 372 2012-06-24T00:00:00 1899-12-30T17:30:00 6139 234 1817 M24 A5A Sunday 196 2012


3180 ENDO12L_M25A MEET-THE-PROFESSOR: CLINICAL - Changing Sex Hormones in Transgender Persons (5:30 PM - 6:15 PM) Changing Sex Hormones in Transgender Persons Wylie C Hembree College of Physicians and Surgeons, Woodcliff Lake, NJ Nothing to Disclose: WCH 2012-06-24T17:30:00 Room 360 2012-06-24T00:00:00 1899-12-30T17:30:00 6170 235 1818 M25 M4A Sunday 197 2012


3181 ENDO12L_M26A MEET-THE-PROFESSOR: CLINICAL - Glucocorticoid-Induced Osteoporosis (GIOP) (5:30 PM - 6:15 PM) Glucocorticoid-Induced Osteoporosis (GIOP) Robert A Adler McGuire Veterans Affairs Medical Center, Richmond, VA Disclosures: RAA: Investigator, Eli Lilly & Company, Merck & Co., Amgen, Novartis Pharmaceuticals; Principal Investigator, Genentech, Inc. 2012-06-24T17:30:00 Room 351 2012-06-24T00:00:00 1899-12-30T17:30:00 6142 236 1819 M26 B3A Sunday 198 2012


3182 ENDO12L_M27A MEET-THE-PROFESSOR: CLINICAL - Growth Hormone Deficiency (5:30 PM - 6:15 PM) Growth Hormone Deficiency Ken Ho Princess Alexandra Hospital, Brisbane, Australia Nothing to Disclose: KH 2012-06-24T17:30:00 Room 370 2012-06-24T00:00:00 1899-12-30T17:30:00 6157 237 1820 M27 PIT2A Sunday 199 2012


3183 ENDO12L_M28A MEET-THE-PROFESSOR: CLINICAL - Klinefelter Syndrome (5:30 PM - 6:15 PM) Klinefelter Syndrome Ronald S Swerdloff LABioMedicine at Harbor UCLA Medical Center, Torrance, CA Nothing to Disclose: RSS 2012-06-24T17:30:00 Room 371 2012-06-24T00:00:00 1899-12-30T17:30:00 6223 238 1821 M28 MR1A Sunday 200 2012


3184 ENDO12L_M29A MEET-THE-PROFESSOR: CLINICAL - Lp(a) [amp] Other CVD Risks: Worth Checking [amp] in Whom? (5:30 PM - 6:15 PM) Lp(a) [amp] Other CVD Risks: Worth Checking [amp] in Whom? Janet B McGill Washington University in St Louis, Saint Louis, MO Nothing to Disclose: JBM 2012-06-24T17:30:00 Room 310 2012-06-24T00:00:00 1899-12-30T17:30:00 6190 239 1822 M29 L3A Sunday 201 2012


3185 ENDO12L_M30A MEET-THE-PROFESSOR: CLINICAL - Medullary Thyroid Cancer (5:30 PM - 6:15 PM) Medullary Thyroid Cancer Richard T Kloos Ohio State University, Columbus, OH Disclosure Incomplete: RTK 2012-06-24T17:30:00 Room 342 ABDE 2012-06-24T00:00:00 1899-12-30T17:30:00 6162 240 1823 M30 TH5A Sunday 202 2012


3186 ENDO12L_M31A MEET-THE-PROFESSOR: CLINICAL - PCOS: Use of Metformin To Allow Pregnancy: Other Treatments (5:30 PM - 6:15 PM) PCOS: Use of Metformin To Allow Pregnancy: Other Treatments Richard S Legro Pennsylvania State University College of Medicine, Hershey, PA Nothing to Disclose: RSL 2012-06-24T17:30:00 Room 352 DEF 2012-06-24T00:00:00 1899-12-30T17:30:00 6149 241 1824 M31 FR1A Sunday 203 2012


3187 ENDO12L_M4A MEET-THE-PROFESSOR: CLINICAL - Pediatric Bone Disease (5:30 PM - 6:15 PM) Pediatric Bone Disease Erik Allen Imel Indiana University School of Medicine, Indianapolis, IN Nothing to Disclose: EAI 2012-06-24T17:30:00 Room 362 2012-06-24T00:00:00 1899-12-30T17:30:00 6246 242 1825 M4 B8A Sunday 204 2012


3188 ENDO12L_M32A MEET-THE-PROFESSOR: CLINICAL - Pediatric Calcium Disorders (5:30 PM - 6:15 PM) Pediatric Calcium Disorders Rachel Ilana Gafni NIH, Bethesda, MD Nothing to Disclose: RIG 2012-06-24T17:30:00 Room 320 2012-06-24T00:00:00 1899-12-30T17:30:00 6214 243 1826 M32 PED6A Sunday 205 2012


3189 ENDO12L_CMF8-1A CASE MANAGEMENT FORUM: CLINICAL - Subtle/Subclinical Hyperthyroidism (1:00 PM - 1:45 PM) Subtle/Subclinical Hyperthyroidism Michael T McDermott University of Colorado Hospital, Denver, CO Nothing to Disclose: MTM 2012-06-25T13:00:00 Theater A 2012-06-25T00:00:00 1899-12-30T13:00:00 6177 283 1942 CMF8-1 CMF-TH1A Monday 255 2012


3190 ENDO12L_CMF8-2A CASE MANAGEMENT FORUM: CLINICAL - Subtle/Subclinical Hyperthyroidism (1:00 PM - 1:45 PM) Subtle/Subclinical Hyperthyroidism Hossein Gharib Mayo Clinic College of Medicine, Rochester, MN Nothing to Disclose: HG 2012-06-25T13:00:00 Theater A 2012-06-25T00:00:00 1899-12-30T13:00:00 6178 283 1943 CMF8-2 CMF-TH1A Monday 256 2012


3191 ENDO12L_M34A MEET-THE-PROFESSOR: CLINICAL - Making Sense of the Numbers: Diagnostic [amp] Treatment Thresholds of Diabetes during Pregnancy (1:00 PM - 1:45 PM) Making Sense of the Numbers: Diagnostic [amp] Treatment Thresholds of Diabetes during Pregnancy Susan Ellen Kirk University of Virginia, Charlottesville, VA Session supported by: Novo Nordisk Inc.[br][br]Nothing to Disclose: SEK 2012-06-25T13:00:00 Room 362 2012-06-25T00:00:00 1899-12-30T13:00:00 6180 284 1944 M34 D6A Monday 257 2012


3255 ENDO12L_M33 MEET-THE-PROFESSOR: CLINICAL - CGMS Sensors: Who Benefits; How [amp] When To Use? (8:30 AM - 9:15 AM) CGMS Sensors: Who Benefits; How & When to Use Joseph Anthony Aloi Eastern Virginia Medical School, Norfolk, VA Session supported by: Dexcom, Inc. [amp] Medtronic Diabetes 2012-06-25T08:30:00 Room 352 DEF 2012-06-25T00:00:00 1899-12-30T08:30:00 6288 251 1835 M33 D3 Monday 33 2012


3088 ENDO12L_S53-3 SYMPOSIUM SESSION: CLINICAL - Hormonal Contraceptives [amp] Replacement Therapy in Adolescents (9:30 AM - 11:00 AM) Reproductive Hormone Use in Anorexia Nervosa [amp] Functional Hypothalamic Amenorrhea: Benefit or Not? Madhusmita Misra Massachusetts General Hospital, Boston, MA Nothing to Disclose: MM 2012-06-26T10:30:00 Room 332 2012-06-26T00:00:00 1899-12-30T10:30:00 6074 368 2695 S53-3 T07-S02 Tuesday 390 2012


3089 ENDO12L_S54-1 SYMPOSIUM SESSION: BASIC - Hormonal Regulation of Primordial Follicle Assembly (9:30 AM - 11:00 AM) A Network of Genes Regulating Primordial Follicle Assembly Eric Eugene Nilsson Washington State University, Pullman, WA The regulation of the process of follicle assembly is of vital importance to the initial formation of the pool of primordial follicles in the ovary. Some growth factors and other gene products have been found to contribute to this regulation. However, it would be desirable to understand the entire network of interacting genes that are involved in follicle assembly. Steps were taken toward developing a systems biology level understanding of gene expression during follicle assembly. Neonatal rat ovaries were treated in culture with one of seven different growth factors known to affect the assembly process (CTGF, AMH FGF2, estradiol, progesterone, activin or TNFa) or were untreated as controls. RNA was collected and subjected to micorarray analysis. Genes differentially expressed between each treatment and controls were identified, and subjected to further network analysis to identify groups of genes co-expressed together. Even though all the treatments affect follicle assembly, there were few genes differentially expressed in common between treatments. A gene bionetwork analysis has detected gene nodules and networks associated with follicle assembly. These gene networks appear critical to determining the primordial follicle pool size. Observations also demonstrate a number of toxicants can environmentally induce epigenetic transgenerational inheritance of ovarian disease with a decline in primordial follicle numbers in the third generation following exposure. This systems level understanding of gene expression during follicle assembly is now being used to investigate the causes of diseases or abnormalities of the follicle pool.[br][br]Sources of Research Support: NIH grants awarded to Michael K. Skinner.[br][br]Nothing to Disclose: EEN 2012-06-26T09:30:00 Room 342 ABDE 2012-06-26T00:00:00 1899-12-30T09:30:00 2517 369 2696 S54-1 T05-S07 Tuesday 392 2012


3090 ENDO12L_S54-2 SYMPOSIUM SESSION: BASIC - Hormonal Regulation of Primordial Follicle Assembly (9:30 AM - 11:00 AM) The Role of BMP Antagonism in Primordial Follicle Assembly Stephanie Ann Pangas Baylor College of Medicine, Houston, TX Genetic or environmental factors that affect the endowment of oocytes or their assembly into the quiescent pool of primordial follicles can disrupt reproductive function and may underlie disorders such as primary ovarian insufficiency. Mouse models have been instrumental in identifying genes important in ovarian development, and a number of genes now associated with ovarian dysfunction in women were first identified as causing reproductive defects in knockout mice. The TGFB family is a large family of developmentally important growth factors that includes TGFBs, anti-Mullerian hormone (AMH), activins, bone morphogenetic proteins (BMPs), and growth and differentiation factor 9 (GDF9). Their activity is negatively regulated by a number of extracellularly secreted antagonists such as follistatin and gremlin, which bind to specific ligands to prevent association and thus, activation of their receptors. Multiple BMP antagonists are expressed in the embryonic and postnatal ovary, but little is known about their roles in ovary development. Loss-of-function of the BMP pathway during early embryogenesis is generally associated with defects in primordial germ cell specification, survival, and migration. We analyzed gain-of-function in the BMP pathway by analyzing mice null for the BMP antagonist, [italic]Grem1[/italic], which is expressed in somatic cells of the ovary. While [italic]Grem1[/italic] knockout (KO) mice die within 24-48 hours of birth due to kidney agenesis, we identified several defects in the ovary of newborn [italic]Grem1[/italic] KO mice. Unexpectedly, deletion of [italic]Grem1[/italic] causes a significant reduction in the number of germ cells, and a delay in primordial follicle assembly. Subsequent analyses have revealed misregulation of additional signal transduction pathways that are known to cause defects in primordial follicle assembly. These data suggest that the BMP pathway has currently uncharacterized roles in regulating oocyte endowment and primordial follicle assembly. These studies will be directly relevant to understanding how disruptions in BMP signaling may result in reproductive diseases in women, and a present a new area for development of tailored diagnostics and interventions.[br][br]Sources of Research Support: Burroughs Wellcome Career Award in the Biomedical Sciences; NIH grant CA138628 from the National Cancer Institute.[br][br]Nothing to Disclose: SAP 2012-06-26T10:00:00 Room 342 ABDE 2012-06-26T00:00:00 1899-12-30T10:00:00 2494 369 2697 S54-2 T05-S07 Tuesday 393 2012


3091 ENDO12L_S54-3 SYMPOSIUM SESSION: BASIC - Hormonal Regulation of Primordial Follicle Assembly (9:30 AM - 11:00 AM) Endocrine Disruption [amp] Primordial Follicle Assembly Wendy N Jefferson NIEHS/NIH, Research Triangle Pk, NC Early in ovarian development, mammalian female germ cells proliferate mitotically but do not undergo complete cytokinesis, resulting in clusters of germ cells called [ldquo]oocyte nests[rdquo]. As the ovary develops, connections between the oocytes are lost and the oocytes become surrounded by somatic pre-granulosa cells, forming primordial follicles. Exposure to estrogenic endocrine disruptors during this critical stage of ovary development has adverse consequences. For example, genistein, a naturally occurring phytoestrogen found in soy products, alters rodent ovarian differentiation and function when exposure occurs in the neonatal period. Female mice treated neonatally with genistein exhibit incomplete oocyte nest breakdown, reduced follicle assembly, and formation of multi-oocyte follicles (MOFs). This effect is mediated by estrogen receptor [beta] (ER[beta]) because ER[beta]-deficient mice do not develop MOFs following genistein treatment. Developmental exposure to other estrogenic compounds such as bisphenol A (BPA) and diethylstilbestrol causes similar ovarian differentiation abnormalities. In pigs, functional consequences of MOFs have been demonstrated because MOFs have altered steroid hormone levels, and oocytes from MOFs are less likely to be fertilized than oocytes from single-oocyte follicles. Follicle assembly in primates occurs prenatally rather than in the postnatal period as in rodents. Blocking estradiol production using an aromatase inhibitor during baboon gestation disrupts follicle assembly, indicating that appropriate levels of estrogen-mediated signaling are required for follicle assembly in primates. Another consequence of inappropriate exposure to estrogenic chemicals is alterations in meiosis during development and in adults. Prenatal BPA exposure in the mouse causes defects in meiosis, and mice lacking ER[beta] have meiotic defects similar to those observed in BPA-treated animals. In the cow, intra-ovarian estrogen levels regulate the timing of oocyte meiotic progression, suggesting that disruption of estrogen signaling in this species might also affect meiosis. There is limited information available in humans, but in a recent study of women undergoing IVF, fewer oocytes were retrieved from women with higher urinary BPA levels. Taken together, these data suggest that inappropriate exposures to estrogenic chemicals may impact ovarian differentiation and/or oocyte development and function later in life.[br][br]Nothing to Disclose: WNJ 2012-06-26T10:30:00 Room 342 ABDE 2012-06-26T00:00:00 1899-12-30T10:30:00 2537 369 2698 S54-3 T05-S07 Tuesday 394 2012


3092 ENDO12L_S55-1 SYMPOSIUM SESSION: TRANSLATIONAL - Mineralocorticoid Hypertension: Advances in Genetics [amp] Management (9:30 AM - 11:00 AM) Clock Genes [amp] Hypertension Hitoshi Okamura Kyoto University, Kyoto, Japan In human, behavior, physiology and metabolism are subject to a well-controlled daily rhythm, generated by an internal self-sustained molecular oscillator called circadian clock. In the present modern society, in which productivity requires around-the-clock activity and demanding shift work, the circadian clock is being compromised. Poor sleep patterns and hectic lifestyle are detrimental to harmonious physiological and metabolic body systems, with severe impact on public health. In our circadian clock system, rhythm is generated by a trillion peripheral cellular clocks throughout the body, supervised by the master clock located in the hypothalamic suprachiasmatic nucleus (SCN), which governs most aspects of physiology and behavior. To exemplify the importance of the biologic clock for health, we used completely arrhythmic mice with the deletion of [italic]Cry1[/italic] and [italic]Cry2[/italic] clock genes ([italic]Cry[/italic]-null mice), in which both master and body clocks are stopped. Recently we found that [italic]Cry[/italic]-null mice showed the salt-sensitive hypertension. Analyzing the adrenal gland by the DNA microarray, we found a novel 3beta-hydroxysteroid dehydrogenase-isomerase (3beta-HSD) gene [italic]Hsd3b6[/italic], which is under clock control, is severely overexpressed specifically in [italic]Cry[/italic]-null mice. [italic]Hsd3b6[/italic] is expressed exclusively in aldosterone-producing cells and is under transcriptional control of the circadian clock. In [italic]Cry[/italic]-null mice, [italic]Hsd3b6[/italic] messenger RNA and protein levels are constitutively high, leading to a marked increase in 3beta-HSD enzymatic activity and, as a consequence, enhanced aldosterone production. These data place [italic]Hsd3b6[/italic] in a pivotal position through which circadian clock malfunction is coupled to the development of hypertension. Translation of these findings to humans will require clinical examination of human [italic]HSD3B1[/italic] gene, which we found to be functionally similar to mouse [italic]Hsd3b6[/italic]. Since HSD3B1 is also an enzyme in the renin-angiotensin-aldosterone system, the characterization of isoenzymes of 3beta-HSD in human hyperaldosteronism will represent a new possibility in the pathogenesis of hypertension in human.[br][br]Nothing to Disclose: HO 2012-06-26T09:30:00 Room 310 2012-06-26T00:00:00 1899-12-30T09:30:00 2417 370 2699 S55-1 T01-S02 Tuesday 396 2012


3093 ENDO12L_S55-2 SYMPOSIUM SESSION: TRANSLATIONAL - Mineralocorticoid Hypertension: Advances in Genetics [amp] Management (9:30 AM - 11:00 AM) Novel Types of Genes in Hyperaldosteronism David S Geller Yale University School of Medicine, New Haven, CT Disclosure Incomplete: DSG 2012-06-26T10:00:00 Room 310 2012-06-26T00:00:00 1899-12-30T10:00:00 6004 370 2700 S55-2 T01-S02 Tuesday 397 2012


3094 ENDO12L_S55-3 SYMPOSIUM SESSION: TRANSLATIONAL - Mineralocorticoid Hypertension: Advances in Genetics [amp] Management (9:30 AM - 11:00 AM) Primary Aldosteronism: Novel Genetics, Novel Therapeutics John W Funder Prince Henry[apos]s Institute of Medical Research, Clayton, Australia Three familial forms of primary aldosteronism (PA) are described to date. FH1 is due to a CYP11BI/CYP11B2 chimera, and very rare; FH2 to an as yet unknown mutation on 7q22; and FH3 to a germline mutation (T158A) in the KCNJ5 [K+] channel. Recently somatic mutations in the KCNJ5 channel (G151R, L168R) have been reported in [sim]40% of aldosterone producing adenomas (APA). Such APA are of a more florid phenotype, occur in younger patients, and have a much higher prevalence in females.Though there are many studies to come, these mutations seem more likely to provide insights into adrenal zonation than therapy. In terms of therapy, PA causes[sim]10% of hypertension; if 20% of the population has high BP, 2% have PA. In no country are [gt]1% of PA patients ever screened, diagnosed or treated. PA has much higher cardiovascular risk than age/sex/BP matched essential hypertension (EH). Low dose MR antagonists are uniquely vasoprotective in EH, and lower BP dramatically in resistant hypertension (BP elevated on 3 conventional antihypertensives including a diuretic). In PA they are treatment of choice in BAH, and longterm may be non-inferior to surgery in APA. Low dose MR antagonists are thus beneficial in EH, and game-changing for the [gt]99% of PA patients who are otherwise never specifically treated. They should therefore be included in first line therapy for all hypertensives; we do not have the resources to diagnose 99% of patients with occult PA, but we do have the means to treat them.[br][br]Nothing to Disclose: JWF 2012-06-26T10:30:00 Room 310 2012-06-26T00:00:00 1899-12-30T10:30:00 2526 370 2701 S55-3 T01-S02 Tuesday 398 2012


3095 ENDO12L_S56-1 SYMPOSIUM SESSION: TRANSLATIONAL - Modulation of the IGF Axis [amp] Cancer (9:30 AM - 11:00 AM) IGFBP-3 [amp] Metastasis Kuk-Wha Lee Mattel Children[apos]s Hospital - UCLA, Los Angeles, CA The insulin-like growth factor binding protein IGFBP-3 is a pro-apoptotic and anti-angiogenic protein in prostate cancer (CaP). Epidemiological studies suggest that low IGFBP-3 is associated with greater risk of aggressive, metastatic prostate cancers, but in vivo functional data are lacking. We demonstrated that mice that are genetically deficient in IGFBP-3 exhibit weaker growth of primary prostate tumors growth but higher incidence of metastatic disease. Prostates in IGFBP-3 knockout mice (IGFBP-3KO mice) failed to undergo apoptosis after castration. Spontaneous prostate tumors did not develop in IGFBP-3KO mice, but splenic lymphomas occurred in 23% of female IGFBP-3KO mice by 80 weeks of age. When we crossed IGFBP-3KO mice with a c-Myc-driven model of CaP that develops slow-growing, non-metastatic tumors; by 24 weeks of age, well-differentiated prostate cancers were observed in all mice regardless of IGFBP-3 status. However, by 80 weeks of age IGFBP-3KO mice tended to exhibit larger prostate tumors than control mice. More strikingly, lung metastases were observed at this time in 55% of the IGFBP-3KO mice but none of the control animals. Cell lines established from Myc:IGFBP-3KO tumors displayed more aggressive phenotypes in proliferation, invasion and colony formation assays, relative to control Myc tumor cell lines. In addition, Myc:IGFBP-3KO cells exhibited evidence of epithelial-mesenchymal transition (EMT). Our findings establish a function for IGFBP-3 in suppressing metastasis in prostate cancer, and they also offer the first reported transgenic model of spontaneous metastatic prostate cancer for studies of this advanced stage of disease.[br][br]Sources of Research Support: NIH grants P50CA92131, P01AG034906, K12HD34610, and DOD award PC061077.[br][br]Nothing to Disclose: K-WL 2012-06-26T09:30:00 Room 370 2012-06-26T00:00:00 1899-12-30T09:30:00 2487 371 2702 S56-1 T06-S02 Tuesday 400 2012


3096 ENDO12L_S56-2 SYMPOSIUM SESSION: TRANSLATIONAL - Modulation of the IGF Axis [amp] Cancer (9:30 AM - 11:00 AM) A Role for Insulin-Like Growth Factor Binding Protein-2 (IGFBP-2) in Cancer Cells Vincenzo Carlo Russo Murdoch Children[apos]s Research Institute, Parkville, Australia The IGF system plays a key role in the regulation of cellular proliferation, differentiation and apoptosis. Disruptions in the balance of IGF system components, including IGFBPs, leading to excessive proliferation and survival signals, have been implicated in the development of many different tumor types.[br]IGFBP-2 is one of the most significant genes in the signature of major cancers including prostate, breast, colon, glioma, adrenocortical, ovarian and lung cancer and paediatric malignancies such as leukaemia, neuroblastoma, astrocytomas. While expression of IGFBP-2 in great excess such as in a transgenic model may lead to IGFBP-2 inhibitory effects via its inhibition of IGF action, local expression by cancers invariably promotes growth and invasion.[br]Although it is clear that IGFBP-2 acts as a modulator of IGF action, with proteolysis of IGFBP-2 further affecting these regulatory interactions, there is now evidence for IGFBP-2 action independent of IGFs. These intrinsic activities of IGFBP-2 do not involve binding to IGFs and are elicited in the absence of IGFs or in the absence of IGF/ IGF-IR signaling. The structural domains involved are likely to be distinct from the IGF-binding sites of IGFBP-2.[br]The mechanisms of IGF-independent actions of IGFBP-2 have been associated with its ability to bind to ECM and the cell surface (e.g. Vitronectin, Integrins, and Proteoglycans) or via intracellular translocation and interaction with proteins such as p21 and PAPA1. More recently we have reported a novel IGF-independent action of IGFBP-2 within the nucleus where IGFBP-2 is involved in the transcriptional activation of VEGF gene. We now have evidence that nuclear transit of IGFBP-2 is mediated by the Importin [alpha] and [beta] complex, interactions occurring via the IGFBP-2 nuclear localization signal (NLS).[br]Whether at the cell surface or intracellular / nuclear, the down-stream events activated by these IGFBP-2 interactions are likely to be crucial to the initiation or enhancement of aggressive behavior in cancer cells.[br]Modulation of IGFBP-2 interactions and / or the associated down-stream events could be exploited to control specific cellular functions altered in cancer cells. However very little has been developed to specifically target IGFBP-2 action in cancer cells and the derived therapies have to date had limited clinical application.[br]The role of IGFBP-2 in cancer cells will be thus reviewed and discussed.[br][br]Nothing to Disclose: VCR 2012-06-26T10:00:00 Room 370 2012-06-26T00:00:00 1899-12-30T10:00:00 2389 371 2703 S56-2 T06-S02 Tuesday 401 2012


3097 ENDO12L_S56-3 SYMPOSIUM SESSION: TRANSLATIONAL - Modulation of the IGF Axis [amp] Cancer (9:30 AM - 11:00 AM) EWS-Fli1 Regulated miRNAs [amp] IGF-I Signaling Paul Jedlicka University of Colorado, Aurora, CO Ewing Sarcoma is an aggressive malignancy with poor long-term outcome. In an effort to identify more effective therapeutic approaches for Ewing Sarcoma, our laboratory has been studying the biology of microRNAs, novel regulators of gene expression with therapeutic potential. As a first approach, we have focused on microRNAs downstream of the EWS/Fli1 fusion oncoprotein, the dominant oncogenic driver in Ewing Sarcoma. Initial studies have identified growth suppressive microRNAs that target the IGF signaling pathway, an important tumor-promoting axis in Ewing Sarcoma. Ongoing studies seek to understand: 1) how these and other microRNAs contribute to Ewing Sarcoma pathogenesis; 2) how the expression of microRNAs is regulated by the EWS/Fli1 oncoprotein; and 3) how these findings could be harnessed for novel therapies for this aggressive disease.[br][br]Sources of Research Support: The Boettcher Foundation[apos]s Webb-Waring Biomedical Research Program, Alex[apos]s Lemonade Stand Foundation for Childhood Cancer, Ruth L. Kirschstein National Research Service Award Institutional Training Grant, and the University of Colorado Cancer Center and School of Medicine.[br][br]Nothing to Disclose: PJ 2012-06-26T10:30:00 Room 370 2012-06-26T00:00:00 1899-12-30T10:30:00 2484 371 2704 S56-3 T06-S02 Tuesday 402 2012


3098 ENDO12L_S57-1 SYMPOSIUM SESSION: TRANSLATIONAL - New Challenges in Obesity Pathogenesis (9:30 AM - 11:00 AM) Insufficient Sleep, Energy Metabolism, and Risk for Obesity Plamen Penev University of Chicago, Chicago, IL Driven by the demands and opportunities of modern life many people now sleep less than six hours per night. In the clinic, this behavior may present as a diagnosis of insufficient sleep syndrome (ICSD-9, #307.49-4) which is receiving increased attention as a potential risk factor for obesity and related metabolic morbidity. A central theme of this presentation will be the notion that extended wakefulness is metabolically costly, which triggers a set of neuroendocrine (increased hunger, reduced satiety), metabolic (lower resting metabolic rate), and behavioral (reduced physical activity) adaptations aimed at increasing food intake and conserving energy. Although this coordinated response may have evolved to offset the metabolic demands of sleeplessness in natural habitats with limited food availability, it can become maladaptive in a modern environment which allows many to overeat while maintaining a sedentary lifestyle without sufficient sleep. Growing experimental evidence now suggests that such sleep-loss-related metabolic adaptation could: 1) lead to increased retention of fat when people aim to return to their usual weight after various life events associated with excessive food intake; and 2) undermine the success of therapies combining reduced food intake and increased physical activity to decrease metabolic risk in obesity-prone individuals. Early observational and clinical field trial data support this experimental framework, making it prudent to recommend that overweight and obese individuals attempting to reduce their caloric intake and maintain increased physical activity should obtain adequate sleep and seek effective treatment for any coexisting sleep disorders.[br][br]Sources of Research Support: NIH grants R01-HL089637, CTSA-RR024999, and P60-DK020595.[br][br]Nothing to Disclose: PP 2012-06-26T09:30:00 Room 362 2012-06-26T00:00:00 1899-12-30T09:30:00 2407 372 2705 S57-1 T04-S01 Tuesday 404 2012


3099 ENDO12L_S57-2 SYMPOSIUM SESSION: TRANSLATIONAL - New Challenges in Obesity Pathogenesis (9:30 AM - 11:00 AM) Weight Gain with Antipsychotic Drug Therapy Peter Manu Hofstra North Shore-Long Island Jewish School of Medicine, Glen Oaks, NY Weight gain leading to obesity is a frequent adverse effect when modern, second-generation antipsychotics are used for the treatment of schizophrenia, bipolar disorder and autism. Drug-induced obesity is considered to have a major pathogenetic contribution to the increased prevalence of insulin resistence, atherogenic dyslipidemia and metabolic syndrome observed in patients with chronic psychotic disorders.[br]This presentation will review first our findings (1) related to the weight-gain nad cardiometabolic risk factors in a group of 338 children and adolescents treated for the first time witn second-generation antipsychotic drugs. After a median of 10.8 weeks of treatment, weight increased by 8.5 kg with olanzapine, 6.1 kg with quetiapine, 5.3 kg with risperidone and 4.4 kg with aripirazole. The untreated control group gained 0.2 kg. Lipid levels (total cholesterol, triglycerides and HDL-cholesterol) worsened significantly during treatment with olanzapine, quetiapine and risperidone, but not with aripiprazole. The data suggest that while all second-generation antipsychotic drugs produce significant weight gain in first-time users, metabolic changes are not uniform for this class of drugs.[br]In the second part of the presentation I will address the issue of obesity as an independent risk factor for coronary heart disease in patients treated with second-generation antipsychotics, based on data comparing the 10-year risk of coronary events in obese (N=44) and normal weight (N=83) patients without metabolic syndrome or current treatment with antihyertensive, hypoglycemic or lipid-lowering drugs (2). The 10-year risk of coronary events was very low and virtually identical in the obese and normal weight subjects (2.3% vs 2.6%), despite excess of 12 BMI units and 16 cm waist circumference in the obese.[br][br](1) Correll CU, Manu P, Olshanskiy V et al. JAMA 2009;302:1765-73. (2) Correll CU, Kane JM, Manu P. Eur Arch Psychiatry Clin Neurosci 2011;261:417-23.[br][br]Nothing to Disclose: PM 2012-06-26T10:00:00 Room 362 2012-06-26T00:00:00 1899-12-30T10:00:00 2422 372 2706 S57-2 T04-S01 Tuesday 405 2012


3100 ENDO12L_S57-3 SYMPOSIUM SESSION: TRANSLATIONAL - New Challenges in Obesity Pathogenesis (9:30 AM - 11:00 AM) The Biology of FTO: Implication for Understanding Common Causes of Obesity Giles Yeo University of Cambridge, Cambridge, UK Sequence variants in the first intron of FTO (Fat mass and Obesity related) are strongly associated with human obesity and carriers of the risk alleles show evidence for increased appetite and food intake. Although global FTO null mice display decreased fat and lean body mass, increased metabolic rate and food intake, this is seen against a complex phenotype of postnatal growth retardation and mortality. In contrast, when we modulated FTO levels discretely in the hypothalamic arcuate nuclei of adult animals, we were able to influence food intake, suggesting tissue specific functions for FTO. FTO is part of the family of Fe(II) 2-oxoglutarate (2-OG) oxygenases and we have shown that FTO can demethylate 3-methylthymine (3meT) and 3-methyluracil (3meU) in vitro, in a 2-OG-dependent manner. However, FTO[apos]s physiological role and how it influences bodyweight is yet to be determined. Using a variety of in vivo, in vitro and biochemical methods, we are currently characterising the molecular mechanism by which FTO controls of energy balance.[br]One approach we have taken is to look for potential protein binding partners of FTO, which may provide us with further clues as to its function. Based on bioinformatic analysis we focused on TRIP4 (Thyroid receptor interacting protein 4). In co-immunoprecipitation experiments, we detect that TRIP4 physically interacts with FTO. We then tested for possible functional interaction of TRIP4 with FTO and found that in a demethylation assay, TRIP4 dose dependently enhances the ability of FTO to demethylate RNA (3meU) but NOT DNA (3meT). Thus we have identified TRIP4 as a novel binding partner for FTO and further study of this little known protein could further illuminate the biology of FTO.[br]In another study using mouse and human cell-lines, we find that both glucose and total amino-acid deprivation regulates FTO expression. In particular, FTO mRNA and protein levels are dramatically down-regulated by total amino-acid deprivation in mouse hypothalamic N46 cells, mouse embryonic fibroblasts (MEFs) and in human HEK293 cells. The drop rate of Fto mRNA is faster than its rate of natural degradation, pointing to regulation at the transcriptional level, which is reversible upon amino-acid replacement. This down-regulation was seen with total amino-acid deficiency, as well as with specific amino acids like glutamine and cysteine/methionine. These data suggest that FTO might play a role in the sensing of amino-acid availability.[br][br]Nothing to Disclose: GY 2012-06-26T10:30:00 Room 362 2012-06-26T00:00:00 1899-12-30T10:30:00 2450 372 2707 S57-3 T04-S01 Tuesday 406 2012


3101 ENDO12L_S58-1 SYMPOSIUM SESSION: CLINICAL - Variability in Vitamin D Assays: Implications for Research [amp] Practice (9:30 AM - 11:00 AM) Pressing Need To Standardize the Vitamin D Assays Ravinder Jit Singh Mayo Clinic, Rochester, MN Recent studies have created a lot of interest in understanding the relevance of Vitamin D in different diseases besides it[apos]s role in rickets and osteomalacia. Thus there has been an increase in testing of Vitamin D in clinical and research laboratories. The results generated by the labs come from various methodologies and have raised some concerns on the quality of the results. Specifically, do all the methods used in different laboratories deliver accurate and quality results? In addition, interpretation of the results have also been confusing as numerous guidelines and recommendations have been made regarding the reference ranges or optimal vitamin D levels, which range from 10-70 ng/ml as minimum optimal level. In circulation, vitamin D is bound to a specific transport protein, vitamin D-binding protein (DBP). In the liver, vitamin D is hydroxylated at the carbon 25-position giving rise to 25-OH vitamin D3, the most abundant circulating form of the vitamin. The final hydroxylation step is catalyzed in the kidney by a 1-a-hydroxylase enzyme, resulting in the production of the biologically active form 1,25-(OH)2-vitamin D. The activity of the renal a-hydroxylase is under tight control so that the production of 1,25-(OH)2-vitamin D remains constant over a wide range of substrate (25-OH-vitamin D) concentrations (4-200 ng/ml). The methodologies including HPLC, RIA with low throughput, automated chemiluminescence immunoassays (high throughput), and liquid chromatography-tandem mass spectrometry (LC-MS/MS) are being used in many clinical laboratories. It is well reported that there are challenges in all of these methods, and high level technical expertise is required to perform the analysis in clinical laboratories. The College of American Pathologists (CAP) and the UK-based DEQAS (Vitamin D external quality assessment scheme) surveys provide independent approaches to monitor the performance of laboratories that use various methods for testing of 25-OH-D. The survey feedback does not assess the accuracy of 25-OH-D measurements by laboratories. CAP data often indicates that clinical laboratories using chemiluminescence immunoassays can report a result for the same sample which is much different as compared to RIA and LC-MS/MS methods. There could be many reasons for these variations, including drifts in the reagents being manufactured, but there is a clear and urgent need for harmonization and standardization.[br][br]Disclosures: RJS: Consultant, Mayo Clinic Patent on Vitamin D Testing. 2012-06-26T09:30:00 Theater A 2012-06-26T00:00:00 1899-12-30T09:30:00 2385 373 2708 S58-1 T03-S02 Tuesday 408 2012


3102 ENDO12L_S58-2 SYMPOSIUM SESSION: CLINICAL - Variability in Vitamin D Assays: Implications for Research [amp] Practice (9:30 AM - 11:00 AM) Implications for Epidemiologic Research Rosemary L Schleicher Centers for Disease Control [amp] Prevention, Atlanta, GA Our laboratory performs 25-hydroxyvitamin D (25OHD) testing for the National Health and Nutrition Examination Survey (NHANES) which provides nationally representative samples of the noninstitutionalized civilian US population. Each year, about 5,000 persons are examined. Biological specimens are collected to assess health and nutrition status. Assessment of vitamin D status has been ongoing since NHANES III (1988-1994); until recently, an RIA was used to measure 25OHD. In the late 1990s, the manufacturer made changes to the assay that affected performance. In the reformulated assay, a higher affinity antibody improved precision and sensitivity by reducing non-specific binding. As shown in later studies, the reformulated assay which was used during NHANES 2000[ndash]2006 measured approximately 12% lower than the original assay used for NHANES 1988[ndash]1994. Several times during analysis of NHANES 2000[ndash]2006 specimens, the assay performed predominantly on one side or the other of long-term QC means thereby affecting reference range and vitamin D deficiency estimates. Using QC materials and RIA data, regression equations were derived to adjust the NHANES 1988[ndash]2006 data to eliminate these assay fluctuations.[br]From NHANES 2007 forward, the laboratory is using a highly accurate isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay to measure serum 25OHD. This assay uses calibration materials that are traceable to standard reference materials (NIST SRM 2972 and 972). A new set of regression equations is being produced to adjust the original NHANES 1988-2006 RIA data, this time to an accurate LC-MS/MS assay. LC-MS/MS-adjusted RIA data will permit comparisons with forthcoming LC-MS/MS data. The C3 epimer of 25OHD[sub]3 [/sub]which appears to lack full biological potential[sub] [/sub]is being chromatographically separated from 25OHD[sub]3 [/sub]in the LC-MS/MS assay. Starting with the 2007-2008 survey, serum concentrations of 25OHD[sub]3[/sub], C3-epi-25OHD[sub]3[/sub] and 25OHD[sub]2[/sub] will be available for the US population. Our lab is participating in a NIH-CDC Vitamin D Standardization Program (VDSP) to standardize 25OHD data from several national health surveys to reference measurement procedures. Accuracy-based external quality assessment programs from NIST and College of American Pathologists have also recently become available. It is expected that 25OHD kit manufacturers will use these new services to better align their methods such that routine clinical assays will be more accurate.[br][br]Nothing to Disclose: RLS 2012-06-26T10:00:00 Theater A 2012-06-26T00:00:00 1899-12-30T10:00:00 2548 373 2709 S58-2 T03-S02 Tuesday 409 2012


3103 ENDO12L_S58-3 SYMPOSIUM SESSION: CLINICAL - Variability in Vitamin D Assays: Implications for Research [amp] Practice (9:30 AM - 11:00 AM) Variability in Vitamin D Assays: Practical Applications [mdash] What Is the Clinician To Do? Marc K Drezner University of Wisconsin School of Medicine Public Health, Madison, WI Although reliable assessment of vitamin D status is measurement of 25-hydroxyvitamin D (25[OH]D), variability in the assay has confounded diagnosis and therapy of vitamin D dependent disorders. Variability is due to differing antibody recognition of vitamin D metabolites, lack of standards for the assays, and co-elution of other vitamin D metabolites with 25(OH)D. This problem is amplified by the recent common practice for laboratories to shift from classical 25(OH)D measurements to more rapid, high-output automated assays, which exhibit inordinate variability. Compounding this problem is interpretation of data in the literature, which has been accrued using various 25(OH)D assays that have drift and shift, and correlation of the reported data with that obtained by the assay in the local laboratory available to the clinician. And, if assay variability is not enough of a problem, the clinician must decide what level of 25(OH)D should prompt therapy for vitamin D insufficiency or deficiency, the dose/frequency of vitamin D to use when therapy is required, and the appropriate time/interval at which to measure 25(OH)D in patients on therapy.[br]To provide guidance regarding what clinicians are to do in order to overcome such problems, these questions will be addressed:[br]1. What serum 25(OH)D level should prompt therapy for vitamin D insufficiency/deficiency or indicate adequate therapy?[br]2. At what intervals should clinicians repeat 25(OH)D measurement to gauge treatement effectiveness?[br]3. What is the optimal 25(OH)D assay(s) for use by a clinician and for use in research studies?[br]4. If the optimal assay is not available to a clinician, what measures should be taken to assure that the assay in use will provide adequate data to serve the needs of the patients?[br]5. Under what circumstances should the clinician require that patient samples are sent to accomplished laboratories, in which a gold standard assay(s) for serum 25(OH)D is performed?[br]While the variability in the measurement of 25(OH)D presents a formidable problem to clinicians, knowledge gleaned, from the answers to the questions posed above, should afford physicians a clear and concise pathway to assure that they are delivering the best care possible, no matter the assay to which they have access. Simply put, the variability can, under most circumstances be reduced to a reasonable limit. Alternatively, when the circumstances demand, commercial availability of the gold standard assays is a viable option.[br][br]Disclosure Incomplete: MKD 2012-06-26T10:30:00 Theater A 2012-06-26T00:00:00 1899-12-30T10:30:00 2467 373 2710 S58-3 T03-S02 Tuesday 410 2012


3105 ENDO12L_CP3 CLINICAL PRACTICE GUIDELINES: CLINICAL - Continuous Glucose Monitoring (11:15 AM - 12:00 PM) Moderator TBD To be determined TBD, TBD Session supported by: Medtronic Diabetes, Merck [amp] Co., Inc. and sanofi-aventis U.S.[br][br]Disclosure Incomplete: Author to be determined 2012-06-26T11:15:00 Grand Ballroom AB 2012-06-26T00:00:00 1899-12-30T11:15:00 6269 385 2777 CP3 CPG3 Tuesday 412 2012


3106 ENDO12L_CDW9-1 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Promotion [amp] Tenure (11:15 AM - 12:15 PM) Promotion [amp] Tenure Simon James Rhodes Indiana University Indianapolis, Indianapolis, IN Session supported by: Novo Nordisk Inc. [br][br]Nothing to Disclose: SJR 2012-06-26T11:15:00 Trainee Career Center 2012-06-26T00:00:00 1899-12-30T11:15:00 6226 386 2778 CDW9-1 CDW6 Tuesday 413 2012


3107 ENDO12L_CDW9-2 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Promotion [amp] Tenure (11:15 AM - 12:15 PM) Promotion [amp] Tenure Sally Ann Camper University Michigan Medical School, Ann Arbor, MI Session supported by: Novo Nordisk Inc. [br][br]Nothing to Disclose: SAC 2012-06-26T11:15:00 Trainee Career Center 2012-06-26T00:00:00 1899-12-30T11:15:00 6227 386 2779 CDW9-2 CDW6 Tuesday 414 2012


3108 ENDO12L_CDW9-3 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Promotion [amp] Tenure (11:15 AM - 12:15 PM) Promotion [amp] Tenure Mark Stephen Roberson Cornell University College of Veterinary Medicine, Ithaca, NY Session supported by: Novo Nordisk Inc. [br][br]Nothing to Disclose: MSR 2012-06-26T11:15:00 Trainee Career Center 2012-06-26T00:00:00 1899-12-30T11:15:00 6228 386 2780 CDW9-3 CDW6 Tuesday 415 2012


3109 ENDO12L_Y3 ENDOCRINE YEAR IN BASIC/CLINICAL SCIENCE: CLINICAL - The Year in Lipids (12:15 PM - 1:00 PM) The Year in Lipids Ira Jay Goldberg Columbia University, New York, NY Session supported by: Merck & Co., Inc.[br][br]Disclosure Incomplete: IJG 2012-06-26T12:15:00 Theater A 2012-06-26T00:00:00 1899-12-30T12:15:00 6117 387 2781 Y3 YI3 Tuesday 416 2012


3110 ENDO12L_CDW10 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Funding [amp] NIH Loan Repayment Opportunities (12:15 PM - 1:00 PM) Funding [amp] NIH Loan Repayment Opportunities Chyren Hunter NIH, Bethesda, MD Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: CH 2012-06-26T12:15:00 Trainee Career Center 2012-06-26T00:00:00 1899-12-30T12:15:00 6229 388 2782 CDW10 CDW7 Tuesday 417 2012


3111 ENDO12L_Y4 ENDOCRINE YEAR IN BASIC/CLINICAL SCIENCE: BASIC - The Year in Obesity [amp] Cancer (1:00 PM - 1:45 PM) The Year in Obesity [amp] Cancer Stephen D Hursting The University of Texas at Austin, Austin, TX Disclosure Incomplete: SDH 2012-06-26T13:00:00 Theater C 2012-06-26T00:00:00 1899-12-30T13:00:00 6118 396 2791 Y4 YI4 Tuesday 418 2012


3112 ENDO12L_CS1 CLARK T SAWIN MEMORIAL LECTURE: TRANSLATIONAL - Nobel Prize Laureates in Endocrinology (2:00 PM - 2:45 PM) Nobel Prize Laureates in Endocrinology Leonard Wartofsky Washington Hospital Center, Washington, DC This Prize Lectureship represents the [bold]Delbert A. Fisher Research Scholar Award[/bold] for 2012 and is presented under the auspices of the Clark T. Sawin Memorial History of Endocrinology Library of The Endocrine Society. The lecture will explore the history of the Nobel Prize in Medicine or Physiology as it relates to prize winners in endocrinology. Some of the questions addressed will include: how many Nobel Prize Medals for Medicine or Physiology have been awarded in endocrinology and to whom? How are Nobel prize awardees selected and by whom? Who received the first Nobel Prize in Medicine? How many awardees have been women endocrinologists? Who was the youngest prize winner? The oldest? Who were sole awardees and who shared the awards? When was the Prize not awarded and why not? Which university spawned the most awardees? Which city? How many awardees had links to The Endocrine Society? And which award was, and still is, surrounded by controversy?[br]In this year[apos]s Sawin History of Endocrinology Lecture, Dr. Wartofsky will present an informative and entertaining overview of the Nobel laureates who worked in endocrinology and the landmark accomplishments that were recognized with this great honor. Since the first Nobel Prize Medals for Medicine or Physiology were awarded in 1901, a significant number of awards have been given for achievements in endocrinology or endocrine physiology. Come and hear little known facts about the awardees and celebrate the success of these outstanding scientists and physicians for their contributions to world health and for the recognition that they brought to our field of endeavor.[br][br]Nothing to Disclose: LW 2012-06-26T14:00:00 Grand Ballroom AB 2012-06-26T00:00:00 1899-12-30T14:00:00 2443 397 2792 CS1 CTS01 Tuesday 419 2012


3113 ENDO12L_M55 MEET-THE-PROFESSOR: CLINICAL - Bone Health Management in Hypothalamic Amenorrhea (2:45 PM - 3:30 PM) Bone Health Management in Hypothalamic Amenorrhea Karen Klahr Miller Massachusetts General Hospital, Boston, MA Nothing to Disclose: KKM 2012-06-26T14:45:00 Room 352 DEF 2012-06-26T00:00:00 1899-12-30T14:45:00 6150 406 2841 M55 FR4 Tuesday 420 2012


3114 ENDO12L_M56 MEET-THE-PROFESSOR: CLINICAL - Ethnic [amp] Racial Differences iin Pathophysiology [amp] Complications of Type 2 Diabetes: Roles [amp] Pitfalls of AIC (2:45 PM - 3:30 PM) Ethnic and Racial Differences in Pathophysiology and Complications of Type 2 Diabetes: Roles and Pitfalls of AIC Kwame Osei Ohio State University Medical Center, Columbus, OH The pathophysiology of type 2 diabetes in African Americans has been recently undergone extensive investigation. African Americans with and without type 2 diabetes are more insulin resistant and hyperinsulinemic when compared to their white counterparts. In addition, beta cell secretory impairment or defects plays critical role in the pathogenesis of type 2 diabetes in African Americans. Therefore, African Americans manifest greater impairment in the ability of beta cell to compensate for insulin resistance as assessed by disposition index (DI) than in white Americans. We have previously suggested that African Americans also manifest greater hepatic insulin resistance and defective hepatic insulin clearance when compared to whites. The reasons for the racial and ethnic differences in insulin sensitivity and beta cell function remain uncertain. In this regard, a major culprit for insulin resistance is intraabdominal visceral adiposity. Indeed, recent investigations have shown that for the same or identical body mass index, African Americans have lower intraabdominal visceral adiposity when compared to whites. In addtion, African Americans have remarkably higher proinflammatory markers such as C-reactive protein (CRP) and oxidized LDL and oxidative stress markers (F2 isoprostanes). However, the role of inflammatory cytokines in the pathogenesis of type 2 diabetes in African American remains uncertain. Furthermore, we have shown that HDL functionality as assesses by paraoxonae1 (PON1) is lower in African Americans than whites. Moreover, the recent observation that African Americans have higher AIC levels raises concerns as to whether there is a phenomenon of universal glycation of all proteins in blacks In summary, there are ethnic and racial differences in the pathogenesis of type 2 diabetes and the associated complications in African Americans and White Americans in USA.. Whether these racial differences are partly responsible for the greeter diabetes - related complication, morbidity and mortality associated with type 2 diabetes in African Americans than whites remain to be investigated.[br][br]Session supported by: Lilly USA, LLC[br][br]Nothing to Disclose: KO 2012-06-26T14:45:00 Room 320 2012-06-26T00:00:00 1899-12-30T14:45:00 2449 407 2842 M56 D7 Tuesday 421 2012


3115 ENDO12L_M57 MEET-THE-PROFESSOR: CLINICAL - Management of Male Hypogonadotropic Hypogonadism through Puberty (2:45 PM - 3:30 PM) Management of Male Hypogonadotropic Hypogonadism through Puberty Erick J Richmond Hospital Nacional de Ninos, San Jose, Costa Rica Nothing to Disclose: EJR 2012-06-26T14:45:00 Room 362 2012-06-26T00:00:00 1899-12-30T14:45:00 6153 408 2843 M57 MR2 Tuesday 422 2012


3116 ENDO12L_M58 MEET-THE-PROFESSOR: CLINICAL - Pediatric Cushing Syndrome (2:45 PM - 3:30 PM) Pediatric Cushing Syndrome Constantine A Stratakis NICHD/NIH, Rockville, MD Disclosure Incomplete: CAS 2012-06-26T14:45:00 Room 351 2012-06-26T00:00:00 1899-12-30T14:45:00 6179 409 2844 M58 A4 Tuesday 423 2012


3117 ENDO12L_M59 MEET-THE-PROFESSOR: CLINICAL - Premenopausal Osteoporosis (2:45 PM - 3:30 PM) Premenopausal Osteoporosis Sue Alison Brown University of Virginia, Charlottesville, VA Nothing to Disclose: SAB 2012-06-26T14:45:00 Room 361 ABDE 2012-06-26T00:00:00 1899-12-30T14:45:00 6245 410 2845 M59 B4 Tuesday 424 2012


3043 ENDO12L_S45-2 SYMPOSIUM SESSION: CLINICAL - Controversial Issues in Cushing[apos]s Syndrome [quot]To Screen or Not To Screen? That Is the Question (3:45 PM - 5:15 PM) How Should We Screen? Niamh Maria Martin Imperial College, London, UK The diagnosis of Cushing[apos]s syndrome may be challenging, particularly in mild disease. Furthermore, the increasing prevalence of obesity and type 2 diabetes, provides considerable overlap between patients with Cushing[apos]s syndrome and the general population. Initial screening tests should have a high sensitivity to correctly identify affected individuals with this rare disease. Unfortunately, due to the trade-off between sensitivity and specificity, this approach may also result in falsely identifying healthy individuals as having Cushing[apos]s syndrome.[br]Current Endocrine Society Clinical Practice Guidelines for the diagnosis of Cushing[apos]s syndrome recommend 24h urine free cortisol (UFC), late-night salivary cortisol, 1mg overnight dexamethasone suppression test (ODT) or standard low dose 48h dexamethasone suppression test (LDDST) as initial first-line screening investigations. UFC measures biologically active unbound circulating cortisol and thus, is not influenced by alterations in cortisol-binding globulin. However, this is frequently a cumbersome and laborious test, often complicated by inadequate urine collection. Loss of diurnal cortisol rhythm is a key feature of Cushing[apos]s syndrome. An elevated sleeping midnight cortisol has reported high sensitivity for the diagnosis of Cushing[apos]s syndrome, although requires an in-patient investigation, limiting its use. Therefore, late-night salivary cortisol measurement is more appealing in terms of ease of collection in the out-patient setting. Unfortunately, analysis of the performance of this test is limited largely by the different assay methodologies used. The use of dexamethasone suppression to confirm Cushing[apos]s syndrome exploits the resistance of the hypothalamo-pituitary-adrenal axis to exogenous glucocorticoids. The ODT is straightforward to perform as an out-patient, whereas the standard LDDST often requires in-patient admission. ODT and LDDST have comparable sensitivities but the LDDST is more specific. Addition of corticotrophin releasing hormone (CRH) on completion of the LDDST has been reported to improve diagnostic accuracy. This has not been borne out by subsequent studies and the role of the dexamethasone suppressed CRH test in the diagnosis of Cushing[apos]s syndrome remains unclear.[br]The best investigative strategy is a combination of tests rather than relying on a single investigation. If the diagnosis is uncertain, it is essential to pause and re-evaluate the patient both clinically and biochemically.[br][br]Sources of Research Support: NM Martin is funded by an NIHR HEFCE Clinical Senior Lecturer Award.[br][br]Nothing to Disclose: NMM 2012-06-25T16:15:00 Theater A 2012-06-25T00:00:00 1899-12-30T16:15:00 171 337 2639 S45-2 T01-S04 Monday 336 2012


3044 ENDO12L_S45-3 SYMPOSIUM SESSION: CLINICAL - Controversial Issues in Cushing[apos]s Syndrome [quot]To Screen or Not To Screen? That Is the Question (3:45 PM - 5:15 PM) Variability in Cushing Syndrome: A Problem? Krystallenia I Alexandraki St Bartholomew[apos]s Hospital, London, UK Significant fluctuations in HPA axis activity may render Cushing[apos]s syndrome (CS) diagnosis problematic. Furthermore, these fluctuations may show a regular periodicity, and in such [apos]cyclical CS[apos] the regular fluctuations of cortisol levels may vary from days to months (1,2). A number of epidemiological studies, small case series and case reports have recently tried to shed light on this ill-defined presentation of CS (2-5).[br]The most widely accepted definition suggests three peaks and two troughs to define cyclicity in CS (2). The problem of such variability in interfering with diagnosis has been recently acknowledged by the Endocrine Society Clinical Practice Guideline for CS, which recommended further evaluation and follow-up after normal or discordant screening tests when there is a clinical suspicion of cyclicity or irregular fluctuations (6).[br]The fluctuations of mean serum cortisol levels, as assessed by a 5-point cortisol day curve, may be used to define variability. Regarding the prevalence of variability, our data suggest the presence of significant variability in 14.9% of patients with Cushing[apos]s disease (CD) and in 10% of patients with ectopic ACTH syndrome (4,5). Within the CS, 54% of patients had CD, 26% ectopic CS, 11% adrenal causes (43% pigmented nodular adrenocortical disease), 9% were not classified (2). In our series true periodicity was rarer, but this has been clearly reported in every type of CS.[br]Various mechanisms have been implicated in the pathogenesis of fluctuations in HPA axis activity, including a possible local origin such as episodic hemorrhage or infarct of the primary tumor, to a hypothalamic origin with varying interplay of neurotransmitters. There has also been speculation that abnormalities of the synchronised Per-Tim pathway in tumor tissue may be responsible.[br]It is evident that diagnostic tests may be difficult to interpret with such inherent variability, and furthermore [apos]cure[apos] and [apos]remission[apos] rates after treatment may be complicated by such inherent changing of HPA axis activity. Occasionally, patients may even cycle from axis over-activity to periods of total cessation leading to an Addissonian crisis as the normal corticotrophs are suppressed.[br]Physicians should be alert to such variability since it can lead to frequent problems in diagnosis and management, and no specific feature has been yet found to be useful as a marker. Understanding the biological basis of this phenomenon will be of great fundamental importance.[br][br](1) Atkinson AB et al., Clin Endocrinol (Oxf) 1992; 36:297. (2) Meinardi JR et al., Eur J Endocrinol. 2007; 157:245. (3) Albiger NM et al., Arq Bras Endocrinol Metabol 2007; 51:1253. (4) Alexandraki KI et al., Eur J Endocrinol. 2009; 160:1011. (5) Isidori AM et al., J Clin Endocrinol Metab 2006; 91:371. (6) Nieman LK et al., J Clin Endocrinol Metab 2008; 93:1526.[br][br]Nothing to Disclose: KIA 2012-06-25T16:45:00 Theater A 2012-06-25T00:00:00 1899-12-30T16:45:00 2531 337 2640 S45-3 T01-S04 Monday 337 2012


3045 ENDO12L_S46-1 SYMPOSIUM SESSION: BASIC - Endocrine Signaling Interrupted (3:45 PM - 5:15 PM) Xenoestrogen Mixture Signaling Via Nongenomic Mechanisms Cheryl S Watson University of Texas Medical Branch, Galveston, TX Xenoestrogens (XEs; nonphysiologic estrogens) contaminate our environment and adversely affect health by imperfectly mimicking endogenous estrogens. They are prevalent in several commercial product use classes including plastics, pesticides, and surfactants. So that their endocrine-disrupting effects can be predicted, prevented, and perhaps remediated, cellular mechanistic explanations for XEs[apos] actions are needed. We have shown that a variety of XEs at very low concentrations (sub-fM-[gt]nM), including a closely structurally related set (the alkylphenols and bisphenol A) potently regulate specific functional responses in pituitary cells by disrupting the nongenomic signaling actions of endogenous estrogens. Physiologic estrogens and XEs stimulate a variety of signaling pathways culminating in integrated activation of downstream mitogen-activated protein kinases (MAPKs) that coordinate many subsequent cellular events. However, since XEs add to the background of endogenous estrogens, and are typically present in the environment in [italic][underline]mixtures[/underline][/italic], we have to understand how and to what extent multiple XEs work together to worsen endocrine disruptions. We have approached this using a pituitary cell culture system optimized for nongenomic responses mediated by membrane estrogen receptors. We systematically challenge the actions of physiologic estrogens with first one, then two, then three XEs. The rapid signaling and functional endpoints that we measure are: the integrated signaling actions on proximal G[sub][alpha]i[/sub] and other second messenger systems, and distal MAPK (ERK, JNKs and p38) activations. The functional endpoints we measure are cell proliferation and prolactin release. We typically observe oscillating time and non-monotonic dose responses, and have probed the complex signaling pathway contributions to these characteristics of the responses. Very weak XEs can enhance the response to endogenous estrogens. As XEs are more efficacious, or are added together, they dramatically attenuate the physiologic estrogen-induced responses, often far below vehicle control levels. We use quantitative high through-put assays to document these results quantitatively. Such predictive patterns for hormonal signaling systems[apos] vulnerability to low concentrations of environmental contaminant mixtures should lead to more rational regulations for permissible XE levels in the environment.[br][br]Sources of Research Support: NIH R01 ES015292 and F31 ES21164; the Passport Foundation.[br][br]Nothing to Disclose: CSW 2012-06-25T15:45:00 Room 361 2012-06-25T00:00:00 1899-12-30T15:45:00 2464 338 2641 S46-1 T08-S06 Monday 339 2012


3046 ENDO12L_S46-2 SYMPOSIUM SESSION: BASIC - Endocrine Signaling Interrupted (3:45 PM - 5:15 PM) Endocrine Disrupter Chemicals Targeting Thyroid Receptors R Thomas Zoeller University of Massachusetts, Amherst, MA Nothing to Disclose: RTZ 2012-06-25T16:15:00 Room 361 2012-06-25T00:00:00 1899-12-30T16:15:00 6086 338 2642 S46-2 T08-S06 Monday 340 2012


3047 ENDO12L_S46-3 SYMPOSIUM SESSION: BASIC - Endocrine Signaling Interrupted (3:45 PM - 5:15 PM) The Impact and Mechanisms of Bisphenol A in the Heart Scott M Belcher University of Cincinnati, Cincinnati, OH Estrogens regulate a variety of physiological, reproductive, and metabolic functions. Estrogen endocrine disrupting chemicals, such as bisphenol A (BPA), influence the hormone[apos]s normal actions to result in complex developmental or physiological phenotypes. Bisphenol A is a high production volume chemical with estrogen-like activities that is used for production of polycarbonate plastic and epoxy resins. At levels commonly observed in humans, BPA can impact estrogen signaling by acting as a partial agonist at nuclear estrogen receptors, and as a full agonist of rapid signaling mechanisms. The results of studies defining the mechanisms responsible for sexually-dimorphic rapid ER-signaling in rodent cardiac myocytes will be presented. Building upon that new mechanistic understanding, the in vivo sex-specific impacts that BPA has on cardiac structure and function were characterized.[br]Design: A life-long dietary exposure model was used with CD1 mice exposed to BPA (0.004-45 mg/kg/day) or 17[alpha]-ethinyl estradiol (EE; 0.02-1.7 [mu]/kg/day). Breeding pairs and resulting offspring were observed from birth through adulthood (12-14 weeks) for alterations in cardiac function. Endpoints related to reproductive maturation and metabolic function were also analyzed to allow assessment of effects on numerous interrelated endocrine endpoints.[br]Results: Disruptive responses to BPA were typically estrogen-like and also observed in at least one of the EE treatment groups. The BPA dose response relationships were non-monotonic with effects most often observed between 0.004 and 4 mg/kg/day. Uterine weight was significantly increased in all EE treatment groups and by 4 mg/kg/day BPA. Bisphenol A-mediated alterations in reproductive maturation and metabolic function were evident in males but not females.[br]Cardiac specific endpoints: In males BPA increased left ventricular wall thickness and increased cardiac fibrosis, without an alteration in myocyte size. A significant decrease in interstitial collagen was observed in females. A model of stress induced cardiac hypertrophy was developed. In males exposed to BPA and EE, a female-like protection to isoproterenol-induced hypertrophy and fibrosis was evident. In contrast, females exposed to BPA exhibited a male-like sensitivity to isoproterenol treatment. Those data suggest that life-long exposure to BPA causes a pathological reversal of normal sexual dimorphism in the heart.[br][br]Sources of Research Support: NIH Grants RC2 ES018765 and R01-ES015145.[br][br]Nothing to Disclose: SMB 2012-06-25T16:45:00 Room 361 2012-06-25T00:00:00 1899-12-30T16:45:00 2439 338 2643 S46-3 T08-S06 Monday 341 2012


3048 ENDO12L_S47-1 SYMPOSIUM SESSION: BASIC - Male Germ Cell Development: The Good, the Bad, [amp] the Ugly (3:45 PM - 5:15 PM) More Than One: Synergistic and Cell Type-Specific Action of TGF[beta] Family Members In Testis Morphogenesis Humphrey Hung-Chang Yao NIEHS/NIH, Research Triangle Park, NC The cascade of morphogenetic events that lead to testis formation in mammals is initiated by the transcription factor SRY (Sex-determining region of the Y chromosome). SRY is expressed exclusively in the Sertoli cell lineage, indicating that the appearance of other somatic cell types and the organization of the testis structure are facilitated by secreted factors produced by SRY-positive Sertoli cells. Indeed, Sertoli cell-derived factors, such as Fibroblast growth factor 9 (FGF9) and Desert hedgehog (DHH), play essential roles in the establishment of the somatic cell lineages and testis cord formation. In search for other secreted factors involved in testis morphogenesis, we have identified unique roles of several members of the transforming growth factor beta (TGF[beta]) family including anti-M[uuml]llerian hormone (AMH), activin B, and activin A. Both AMH and activin B are produced by Sertoli cells. Loss of either Amh or activin B did not affect organization of the testis. However in the absence of both Amh and activin B, Sertoli cells underwent transdifferentiation into granulosa cells and testis structures degenerated. Activin A, on the other hand, is a product of fetal Leydig cells. Leydig cell-specific ablation of activin A led to decreased Sertoli cell proliferation and stunted testis cord elongation. These data reveal stage- and cell type-specific roles of these these TGF[beta] proteins. Sertoli cell-derived AMH and activin B function as autocrine factors that maintain Sertoli cell fate. When both Amh and activin B are absent, Sertoli cells lose their identity and transdifferentiate into granulosa cells. Activin A from the Leydig cells act as a paracrine factor on Sertoli cells to ensure proper expansion of the testis cords. It is intriguing how these TGF[beta] proteins, despite their redundancy and synergism in intracellular signal transduction, elicit unique actions during testis morphogenesis. Challenges await us in deciphering how these factors interact and regulate their specific downstream cellular events in various somatic cell lineages in the testis.[br][br]I would like to acknowledge Heather Franco, Chia-Feng Liu, and Denise Archambeault, who contributed to the findings described in this work.[br][br]Sources of Research Support: This study was supported by NIH-HD046861 and the NIH Intramural Research Program.[br][br]Nothing to Disclose: HH-CY 2012-06-25T15:45:00 Room 342 ABDE 2012-06-25T00:00:00 1899-12-30T15:45:00 2408 339 2644 S47-1 T05-S08 Monday 343 2012


3049 ENDO12L_S47-2 SYMPOSIUM SESSION: BASIC - Male Germ Cell Development: The Good, the Bad, [amp] the Ugly (3:45 PM - 5:15 PM) Androgen Receptor Mediated Autocrine [amp] Paracrine Signaling in Testis Lee B Smith The University of Edinburgh, Edinburgh, UK Nothing to Disclose: LBS 2012-06-25T16:15:00 Room 342 ABDE 2012-06-25T00:00:00 1899-12-30T16:15:00 6061 339 2645 S47-2 T05-S08 Monday 344 2012


3050 ENDO12L_S47-3 SYMPOSIUM SESSION: BASIC - Male Germ Cell Development: The Good, the Bad, [amp] the Ugly (3:45 PM - 5:15 PM) Regulation of Post-Testicular Sperm Maturation Ilpo T Huhtaniemi Imperial College of London, London, UK The epididymal transfer is vital for the acquisition of sperm motility and fertilizing capacity. Therefore the epididymis is an excellent target for male contraception, also because many of its functions are regulated by epididymis-specific genes. An effective way to explore the function of epididymal genes is their knockout (KO) followed by assessment of the consequences on male fertility. We have recently produced several epididymis-specific KO mice. Targeted disruption of the novel gene, Rnase10, encoding a secreted proximal epididymal protein, resulted in male subfertility associated with an attachment defect of spermatozoa and inability to pass through the female utero-tubal junction. The KO spermatozoa presented with gradual loss of the ADAM3 protein from sperm surface during their epididymal transit, lacked calcium-dependent association with the glutinous extra-cellular material, and were released as single vigorously motile cells into bicarbonate-free medium. They displayed no tendency for head-to-head agglutination and lacked the affinity to the oviductal epithelium and zona pellucida, yet they were capable of fertilization. Although the loss of fertility was partial our findings prompt the study of effects of the human othologue of Rnase10 on male fertility. Another KO we recently produced was that of the androgen receptor (AR) in the proximal epididymis. This model was expected to answer the question whether the epididymal initial segment is a direct androgen target. We produced mice carrying a conditional ARloxP allele together with the Cre recombinase driven by the Rnase10 promoter. Prepubertally, upon the onset of Rnase10 expression, AR became selectively inactivated in principal cells of the proximal epididymis, resulting in epithelial hypoplasia and hypotrophy, followed by epididymal obstruction and development of spermatic granulomata, back pressure-induced atrophy of the seminiferous epithelium, orchitis, and fibrosis of the testicular parenchyma. When the effect of this AR KO was compared with that of efferent duct ligation and orchidectomy, we identified genes specifically regulated by androgen, by testicular efferent fluid and by both. These findings demonstrate that the development and function of the epididymal initial segment is critically dependent on direct androgen regulation, and provided a novel model for obstructive azoospermia. The downstream genes from AR may prove targets for the manipulation of epididymal sperm maturation.[br][br]Sources of Research Support: Wellcome Trust Grant 086695/Z/08/Z.[br][br]Nothing to Disclose: ITH 2012-06-25T16:45:00 Room 342 ABDE 2012-06-25T00:00:00 1899-12-30T16:45:00 2420 339 2646 S47-3 T05-S08 Monday 345 2012


3051 ENDO12L_S48-1 SYMPOSIUM SESSION: TRANSLATIONAL - Neuro-Modulatory Regulation of Gonadotropin Release: NKB (3:45 PM - 5:15 PM) NKB [amp] NK3R Mutations in Humans A Kemal Topaloglu Cukurova University, Faculty of Medicine, Balcali, Adana, Turkey Recent identification of inactivating TAC3 or TACR3 (encoding neurokinin B and its receptor, NK3R, respectively) mutations as the causes of complete normosmic idiopathic hypogonadotropic hypogonadism (IHH) has provided compelling evidence for the involvement of neurokinin B (NKB) signaling in human puberty and reproduction. To date more than 20 mutations in TACR3 and four mutations in TAC3 have been reported. Screening of large cohorts of normosmic IHH revealed a frequency of about 5% for TACR3 mutations. With the absence of animal models in which to explore the exact consequences of NKB signaling dysfunction, studying these human cases have been of great help in identifying the roles of NKB signaling in the hypothalamo pituitary gonadal axis. These studies indicated that NKB signaling acts proximally in an autocrine or paracrine manner to kisspeptin signaling, which elicits GNRH release. Almost invariably affected male infants have micropenis suggesting an essential role for NKB signaling also for in utero activation of the hypothalamo pituitary gonadal axis.[br][br]Nothing to Disclose: AKT 2012-06-25T15:45:00 Room 310 2012-06-25T00:00:00 1899-12-30T15:45:00 2516 340 2647 S48-1 T06-S05 Monday 347 2012


3052 ENDO12L_S48-2 SYMPOSIUM SESSION: TRANSLATIONAL - Neuro-Modulatory Regulation of Gonadotropin Release: NKB (3:45 PM - 5:15 PM) Role of Hypothalamic NKB Neurons in the Physiology of Postmenopausal Women Naomi E Rance University of Arizona College of Medicine, Tucson, AZ Human menopause is characterized by ovarian failure, gonadotropin hypersecretion and dramatic changes in neuronal morphology and neuropeptide gene expression in the infundibular (arcuate) nucleus of the hypothalamus. In postmenopausal women, a subpopulation of neurons expressing estrogen receptor alpha, neurokinin B (NKB), kisspeptin and dynorphin undergo hypertrophy, and exhibit increased NKB and kisspeptin gene expression. Numerous studies have shown that the increase in NKB and kisspeptin gene expression in postmenopausal women is secondary to the loss of ovarian estrogen. A homologous group of kisspeptin/neurokinin B/dynorphin (KNDy) neurons resides in the arcuate nucleus of a variety of other species, including mice, rats, sheep, goats and monkeys. In the rat, KNDy neurons express NK[sub]3[/sub]R and form an interconnected network within the arcuate nucleus projecting to GnRH terminals in the median eminence. This network provides an anatomical framework to explain how coordination among these neurons could mediate feedback information from the gonads to modulate pulsatile GnRH secretion. Because of the diverse projections of KDNY neurons to multiple homeostatic and neuroendocrine control centers, these neurons could integrate sex-steroid signals with a variety of physiological systems. The close association of LH pulses with hot flushes in women has led us to hypothesize that arcuate NKB neurons play a role in the generation of menopausal flushes. In support of this hypothesis, focal microinfusion of the NK[sub]3[/sub]R agonist, senktide, reduces core temperature and activates the median preoptic nucleus, a key component of the heat defense pathway. These data provide the first evidence of NKB signaling in the central control of body temperature. To further investigate the function of KNDy neurons, we developed a method to ablate these cells using a selective NK[sub]3[/sub]R-agonist conjugated to the ribosome-inactivating toxin, saporin. These studies have provided evidence that arcuate KNDy neurons are essential for tonic gonadotropin secretion, the rise in LH after ovariectomy and the estrogen modulation of body weight. We are currently determining the effects of KNDy neuron ablation on the estrogen modulation of thermoregulation. Studies of the postmenopausal human hypothalamus have provided novel information on the neuroendocrine control of reproduction and may even provide clues to the etiology of hot flushes.[br][br]Sources of Research Support: NIH National Institute on Aging R01 AG032315.[br][br]Nothing to Disclose: NER 2012-06-25T16:15:00 Room 310 2012-06-25T00:00:00 1899-12-30T16:15:00 2435 340 2648 S48-2 T06-S05 Monday 348 2012


3053 ENDO12L_S48-3 SYMPOSIUM SESSION: TRANSLATIONAL - Neuro-Modulatory Regulation of Gonadotropin Release: NKB (3:45 PM - 5:15 PM) NKB [amp] the Regulation of LH Secretion in Sheep Robert L Goodman West VA University Health Science Center, Morgantown, WV Disclosure Incomplete: RLG 2012-06-25T16:45:00 Room 310 2012-06-25T00:00:00 1899-12-30T16:45:00 6068 340 2649 S48-3 T06-S05 Monday 349 2012


3054 ENDO12L_S49-1 SYMPOSIUM SESSION: BASIC - Putting the Brakes on Breast Cancer (3:45 PM - 5:15 PM) microRNAs and Hormone Receptor Action in Breast Cancer Jennifer K Richer University of Colorado Anschutz, Aurora, CO The microRNA (miRNA) families miR-200 and miR-221 are the most differentially expressed in estrogen receptor alpha (ER) positive versus negative breast cancers (BrCa). These two families have dueling effects on epithelial cell identity. The human RNaseIII-type nuclease, Dicer, which performs the final step of miRNA biogenesis, is inversely controlled by these two miRNA families. Indeed, we demonstrate that Dicer protein is expressed at four-fold lower levels in TNBC versus adjacent normal breast epithelium or ER+ tumors. We propose that lower Dicer results in reduced overall miRNA expression and the less differentiated state of TNBC. Our group and others have identified miRNAs regulated by ER. To identify PR-regulated miRNAs, we performed gene expression profiling for mature miRNAs upon progestin treatment of luminal BrCa cells. Twenty-eight miRNAs were significantly altered following 6 hrs of progestin treatment. Many known progestin-responsive genes are putative targets of the progestin-regulated miRNAs. As specific examples, we demonstrate that progestin-mediated downregulation of miR-29 relieves repression of two PR-driven genes ATPase, Na[sup]+[/sup]/K[sup]+[/sup] transporting beta 1 ([italic]ATP1B1[/italic]) and Kruppel-like factor 4 [italic](KLF4)[/italic]. We also demonstrate that PR itself is directly targeted by a progestin-upregulated miRNA, miR-513a-5p. This miRNA limits the amount of PR induced by estradiol treatment in multiple luminal BrCa cell lines, providing a novel mechanism for tight control of PR protein levels. Lastly, we demonstrate that an estrogen-regulated miRNA, miR-26, directly targets PR and is responsible for cell-type specific expression of PR in normal cycling mouse and human uteri and in models of endometrial cancer. In summary, miRNAs regulated by steroid hormone receptors (SHR) can target genes that the receptors regulate at the transcriptional level via their classical action as transcription factors. We provide specific examples of PR-regulated genes controlled at both the 5[apos] regulatory regions by PR-mediated transcription and at the 3[apos]UTR by PR-regulated miRNAs that control translation. Such dual hormone-mediated regulation provides an additional level of control that allows fine tuning of the timing and even cell-type specificity of hormone-responsive proteins. Collectively, our data demonstrate that miRNAs serve as important mediators of SHR expression and action.[br][br]Sources of Research Support: DOD Breast Cancer Program Idea Award BC074403,Avon Breast Cancer Foundation for Women, and NIH NICHD P01 PAR-10-245 project 3 to JKR.[br][br]Nothing to Disclose: JKR 2012-06-25T15:45:00 Room 370 2012-06-25T00:00:00 1899-12-30T15:45:00 2519 341 2650 S49-1 T02-S07 Monday 351 2012


3055 ENDO12L_S49-2 SYMPOSIUM SESSION: BASIC - Putting the Brakes on Breast Cancer (3:45 PM - 5:15 PM) Dietary Targeting of Tumor Suppressors and Oncogenes for Breast Cancer Prevention Rosalia C M Simmen University of Arkansas for Medical Science [amp] Arkansas Children[apos]s Nutrition Center, Little Rock, AR Breast cancer is a complex disease that arises from genetic and epigenetic changes in molecules that are critical for growth control, DNA repair, apoptosis and differentiation. The incidence of breast cancer varies worldwide, implicating diet and lifestyle disparities among the general population as also contributory to breast cancer development and progression. Diet-mediated changes in transcriptional programs that promote the early differentiation of the mammary gland may lead to reduced breast cancer risk. Our analyses of rodent models of breast cancer and mammary epithelial and stromal fibroblast cell lines have provided significant insights into the developmental context, target cells, mechanisms of actions, and nature of bioactive components for breast cancer prevention by diet. One example is the soy isoflavone genistein that targets mammary epithelial cells and neighboring stromal adipocytes via estrogen receptor (ER)-dependent and -independent mechanisms to promote mammary epithelial differentiation. We observed that the mammary basal and luminal epithelial sub-populations (termed stem-like/progenitor cells) that are able to self-renew, grow as spheroids in the absence of attachment, and seed tumors when dys-regulated, are also targeted by certain dietary factors. Genome-wide profiling of mammary epithelial and basal stem-like epithelial subpopulations as a function of diet revealed signaling pathways involved in immune response, protein and carbohydrate metabolism, growth regulation and stem cell niche as potentially contributing to increased resistance to carcinogenic insult. The promotion of differentiation and reduction in proliferative indices of mammary epithelial cells were accompanied by increased tumor suppressors PTEN and p53 expression, enhanced E-cadherin membrane association, and attenuated [beta]-catenin nuclear localization. In stromal adipocytes, increased adipokine adiponectin secretion and higher ER-[beta] expression constitute favorable outcomes of the mammary tumor protective effects of dietary factors. Mammary tumor prevention conferred by dietary factors is dependent on developmental context, with in utero/lactational exposure via maternal diet providing an early window for nutritional targeting. Understanding the gene and cell regulatory programs directed by bioactive dietary factors in the distinct mammary cell compartments has important implications for the development of novel strategies for breast cancer prevention and treatment.[br][br]Sources of Research Support: USDA-CRIS-6251-5100002-06S; Arkansas Breast Cancer Research Program; UAMS Translational Research Institute (CTSA Award #1UL1RR029884), Department of Defense Breast Cancer Program.[br][br]Nothing to Disclose: RCMS 2012-06-25T16:15:00 Room 370 2012-06-25T00:00:00 1899-12-30T16:15:00 2395 341 2651 S49-2 T02-S07 Monday 352 2012


3056 ENDO12L_S49-3 SYMPOSIUM SESSION: BASIC - Putting the Brakes on Breast Cancer (3:45 PM - 5:15 PM) Forkhead Transcription Factor Control of Breast Epithelial Cell Phenotypes Ruth Ann Keri Case Western Reserve University School of Medicine, Cleveland, OH Breast cancer is a heterogeneous disease whose subtypes have been defined by gene expression profiling. Expression signatures of these subtypes overlap with distinct cell types during mammary morphogenesis, with luminal tumors corresponding to mature luminal cells and the basal, more aggressive subtype being similar to luminal progenitors. This suggests either that the signatures are reflective of the tumor cells-of-origin or that oncogenic events induce profiles that are similar to developmental stages. Luminal breast cancers consistently express the transcription factors FOXA1, estrogen receptor-[alpha] (ER), and GATA-3 that comprise a regulatory network that controls growth and differentiation of luminal breast cancers. FOXA1 is essential for mediating responsiveness to estrogens and this has is purported to be due to a pioneering function of FOXA1 on ER target genes. Using multiple transplantation paradigms of FOXA1 null mammary glands, we reported that FOXA1 is critical for ductal elongation, but is dispensable for alveologenesis. We also found that FOXA1 binds to, and is necessary for, ER expression. These results indicate that FOXA1 regulates estrogen responsiveness of mammary epithelial cells through multiple mechanisms including collaborating with ER to induce target gene expression as well as stimulating expression of ER itself. FOXA1 also has ER-independent functions in breast cancer. FOXA1 represses expression of basal subtype-associated genes, in part by directly binding to these genes. Hence, FOXA1 both promotes the luminal gene signature while repressing the profile associated with basal breast cancer cells. Transient silencing of FOXA1 in luminal cells results in increased migration and invasion, two properties of basal type breast cancers, indicating that FOXA1 regulation of basal-type genes is functionally significant. Like luminal cancers, basal breast tumors also express signature transcription factors. FOXC1 is highly expressed in basal subtype tumors and its expression is diametrically opposed to FOXA1. Although we found that FOXC1 is not necessary for normal mammary morphogenesis, our studies have revealed that it is a key regulator of invasion of breast cancer cell lines. As a whole, our studies reveal that the two forkhead box transcription factors, FOXA1 and FOXC1, play opposing roles in dictating luminal and basal breast cancer phenotypes that ultimately control disease outcome.[br][br]Sources of Research Support: Department of Defense Grants W81XWH-06-1-0712, W81XWH-09-1-0696, and W81XWH-08-1-0347, and NIH Grant R01 CA090398.[br][br]Nothing to Disclose: RAK 2012-06-25T16:45:00 Room 370 2012-06-25T00:00:00 1899-12-30T16:45:00 2497 341 2652 S49-3 T02-S07 Monday 353 2012


3057 ENDO12L_L6-1 PLENARY: TRANSLATIONAL - Aging of the Female Brain: Phenotypes of Vulnerability [amp] Targets of Opportunity To Prevent Neurodegenerative Disease (5:30 PM - 6:30 PM) Aging of the Female Brain: Phenotypes of Vulnerability [amp] Targets of Opportunity To Prevent Neurodegenerative Disease Roberta Diaz Brinton University of Southern California, Los Angeles, CA Disclosure Incomplete: RDB 2012-06-25T17:30:00 Grand Ballroom AB 2012-06-25T00:00:00 1899-12-30T17:30:00 6126 350 2662 L6-1 PL06-1 Monday 354 2012


3058 ENDO12L_L6-2 PLENARY: TRANSLATIONAL - Discovering a Drug: The Big Picture (5:30 PM - 6:30 PM) Discovering a Drug: The Big Picture Michael Rosenblatt Merck [amp] Company, Whitehouse Station, NJ New drugs and vaccines are not [quot]discovered[quot] -- they are invented. They are generated by pharmaceutical and biotechnology companies. These are the only organizations that can go all the way from drug discovery to clinical trials, to worldwide distribution. Pharmaceutical and biotechnology companies are positioned in an interactive ecosystem that includes academia and government. The process of bringing a drug from concept to new therapy has the longest product cycle of any industry, and it occurs in the context of the most regulated industry in the world. The elapsed time from bench to bedside is 10-15 years with an associated cost of $1-2 billion per new drug. Despite the enormous effort and expenditure, the probability of success is very low. The business model for innovation is increasingly challenged. Challenges facing the industry include ballooning costs for basic science (e.g. genomics, imaging, technologies), patent expiry, pressures on pricing, generic competition (created by the previous successes of the research-driven pharmaceutical industry), payer and policymaker mandates on what physicians can prescribe, comparative effectiveness testing, and post-approval regulation. The public health benefit of vaccines and drugs for infectious disease, diabetes, cardiovascular disease, osteoporosis, respiratory disease, reproduction, cancer and other medical needs has been substantial on life expectancy and quality of life. But there remain many unmet medical needs and patient access remains a problem. Major geographies in the world have only recently begun to experience the impact of modern medicines and vaccines.[br]Enhanced collaboration (appropriately constructed) between academia, industry, and government in drug discovery, clinical trials, health information technology and global health have the potential to speed and increase advances in health.[br][br]Disclosures: MR: Employee, Merck & Co.; Founder, Radius Pharmaceutical, Theracrine. 2012-06-25T17:30:00 Grand Ballroom AB 2012-06-25T00:00:00 1899-12-30T17:30:00 2431 351 2663 L6-2 PL06-2 Monday 355 2012


3059 ENDO12L_CDW7-1 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - How To Write a Successful Career Development Award Application (7:00 PM - 9:00 PM) How To Write a Successful Career Development Award Application Ellen W Seely Brigham [amp] Women[apos]s Hospital, Boston, MA Session supported by: Novo Nordisk Inc. [br][br]Nothing to Disclose: EWS 2012-06-25T19:00:00 Hilton Americas-Houston, Ballroom of the Americas A 2012-06-25T00:00:00 1899-12-30T19:00:00 6224 354 2666 CDW7-1 CDW5 Monday 356 2012


3060 ENDO12L_CDW7-2 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - How To Write a Successful Career Development Award Application (7:00 PM - 9:00 PM) How To Write a Successful Career Development Award Application Marie Catherine Gelato SUNY Health Science Center, Stony Brook, NY Session supported by: Novo Nordisk Inc. [br][br]Nothing to Disclose: MCG 2012-06-25T19:00:00 Hilton Americas-Houston, Ballroom of the Americas A 2012-06-25T00:00:00 1899-12-30T19:00:00 6225 354 2667 CDW7-2 CDW5 Monday 357 2012


3061 ENDO12L_CDW7-3 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - How To Write a Successful Career Development Award Application (7:00 PM - 9:00 PM) How To Write a Successful Career Development Award Application Joan M Lakoski University of Pittsburgh Health Science, Pittsburgh, PA Session supported by: Novo Nordisk Inc. [br][br]Nothing to Disclose: JML 2012-06-25T19:00:00 Hilton Americas-Houston, Ballroom of the Americas A 2012-06-25T00:00:00 1899-12-30T19:00:00 6255 354 2668 CDW7-3 CDW5 Monday 358 2012


3062 ENDO12L_CDW7-4 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - How To Write a Successful Career Development Award Application (7:00 PM - 9:00 PM) How To Write a Successful Career Development Award Application Dolores M Shoback UCSF/VA Medical Center, San Francisco, CA Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: DMS 2012-06-25T19:00:00 Hilton Americas-Houston, Ballroom of the Americas A 2012-06-25T00:00:00 1899-12-30T19:00:00 6256 354 2669 CDW7-4 CDW5 Monday 359 2012


3063 ENDO12L_CDW7-5 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - How To Write a Successful Career Development Award Application (7:00 PM - 9:00 PM) How To Write a Successful Career Development Award Application Jonathan S Williams Brigham [amp] Women[apos]s Hospital, Boston, MA Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: JSW 2012-06-25T19:00:00 Hilton Americas-Houston, Ballroom of the Americas A 2012-06-25T00:00:00 1899-12-30T19:00:00 6257 354 2670 CDW7-5 CDW5 Monday 360 2012


3064 ENDO12L_CDW7-6 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - How To Write a Successful Career Development Award Application (7:00 PM - 9:00 PM) How To Write a Successful Career Development Award Application Joy Y Wu Endocrine Unit, Boston, MA Session supported by: Novo Nordisk Inc. [br][br]Nothing to Disclose: JYW 2012-06-25T19:00:00 Hilton Americas-Houston, Ballroom of the Americas A 2012-06-25T00:00:00 1899-12-30T19:00:00 6259 354 2671 CDW7-6 CDW5 Monday 361 2012


3065 ENDO12L_CDW7-7 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - How To Write a Successful Career Development Award Application (7:00 PM - 9:00 PM) How To Write a Successful Career Development Award Application John F Connaughton NIH/NIDDK, Bethesda, MD Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: Author to be determined 2012-06-25T19:00:00 Hilton Americas-Houston, Ballroom of the Americas A 2012-06-25T00:00:00 1899-12-30T19:00:00 6283 354 2672 CDW7-7 CDW5 Monday 362 2012


3066 ENDO12L_CDW8 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Writing a Successful R01 [amp] Other Independent Research Grant Applications (7:00 PM - 9:00 PM) Writings a Successful R01 [amp] Other Independent Research Grant Applications Ronald N Margolis NIDDK/NIH, Bethesda, MD Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: RNM 2012-06-25T19:00:00 Hilton Americas-Houston, Ballroom of the Americas C 2012-06-25T00:00:00 1899-12-30T19:00:00 6252 355 2673 CDW8 CDW9 Monday 364 2012


3067 ENDO12L_L7-1 PLENARY: TRANSLATIONAL - Edwin B. Astwood Award Lecture: Combinatoriality, Signal Integration [amp] Allostery: Determinants of Specific Transcriptional Regulation by the Glucocorticoid Receptor (8:00 AM - 9:15 AM) Edwin B. Astwood Award Lecture: Combinatoriality, Signal Integration [amp] Allostery: Determinants of Specific Transcriptional Regulation by the Glucocorticoid Receptor Keith R Yamamoto University of California- San Francisco, San Francisco, CA Nothing to Disclose: KRY 2012-06-26T08:00:00 Grand Ballroom AB 2012-06-26T00:00:00 1899-12-30T08:00:00 6127 356 2674 L7-1 PL07-1 Tuesday 365 2012


3068 ENDO12L_L7-2 PLENARY: TRANSLATIONAL - Cell Selective Interaction of Transcription Factors with Chromatin (8:00 AM - 9:15 AM) Cell Selective Interaction of Transcription Factors with Chromatin Gordon L Hager National Institute of Health, Bethesda, MD Nothing to Disclose: GLH 2012-06-26T08:00:00 Grand Ballroom AB 2012-06-26T00:00:00 1899-12-30T08:00:00 6128 357 2675 L7-2 PL07-2 Tuesday 366 2012


3069 ENDO12L_CMF10-1 CASE MANAGEMENT FORUM: CLINICAL - Management of Pediatric Thyroid Cancer (8:30 AM - 9:15 AM) Management of Pediatric Thyroid Cancer Catherine Anne Dinauer Yale University School of Medicine, Guilford, CT Session supported by: Genzyme Corporation[br][br]Nothing to Disclose: CAD 2012-06-26T08:30:00 Theater B 2012-06-26T00:00:00 1899-12-30T08:30:00 6185 359 2677 CMF10-1 CMF-TH2 Tuesday 367 2012


3070 ENDO12L_CMF10-2 CASE MANAGEMENT FORUM: CLINICAL - Management of Pediatric Thyroid Cancer (8:30 AM - 9:15 AM) Management of Pediatric Thyroid Cancer Gary Lee Francis Medical College of Virginia, Richmond, VA Session supported by: Genzyme Corporation[br][br]Nothing to Disclose: GLF 2012-06-26T08:30:00 Theater B 2012-06-26T00:00:00 1899-12-30T08:30:00 6186 359 2678 CMF10-2 CMF-TH2 Tuesday 368 2012


3071 ENDO12L_M49 MEET-THE-PROFESSOR: CLINICAL - Acromegaly (8:30 AM - 9:15 AM) Acromegaly Andrea Lynn Utz Vanderbilt University, Nashville, TN Session supported by: Ipsen Biopharmaceuticals, Inc.[br][br]Disclosures: ALU: Advisory Group Member, Ipsen. 2012-06-26T08:30:00 Room 310 2012-06-26T00:00:00 1899-12-30T08:30:00 6165 360 2679 M49 PIT4 Tuesday 369 2012


3118 ENDO12L_PS8-1 PRESIDENTIAL SYMPOSIUM SESSION: CLINICAL - Diabetes [amp] The Brain (3:45 PM - 5:15 PM) Hypoglycemia Elizabeth Rachel Seaquist University of Minnesota, Minneapolis, MN Nothing to Disclose: ERS 2012-06-26T15:45:00 Grand Ballroom AB 2012-06-26T00:00:00 1899-12-30T15:45:00 6130 411 2846 PS8-1 SSS06 Tuesday 426 2012


3119 ENDO12L_PS8-2 PRESIDENTIAL SYMPOSIUM SESSION: CLINICAL - Diabetes [amp] The Brain (3:45 PM - 5:15 PM) Chronic Hyperglycemia and Its Impact on Brain Function and Structure Christopher M Ryan University of Pittsburgh, Pittsburgh, PA Early neuropathology studies of brain autopsy material indicated that poorly controlled diabetes can induce a profound diabetic encephalopathy. More recent reports have failed to find evidence of such extreme functional or structural changes, but a growing literature has found compelling evidence that diabetes and its treatment can disrupt the integrity of the CNS. In this presentation I argue that distinct [apos]neurocognitive phenotypes[apos] can be identified for children and adults with type 1 or type 2 diabetes, that these may reflect somewhat different pathophysiological processes, and these have different implications for the remediation and prevention of neurocognitive complications. Studies of adults with type 1 diabetes have identified a potent predictor of brain dysfunction: microvascular damage secondary to chronic hyperglycemia. Patients with retinopathy and other clinically significant complications have an increased risk of manifesting reductions in gray and white matter brain volume, and are more likely to show cognitive sequela characterized by poorer performance on tasks that require rapid information-processing, sustained attention, and verbal intelligence. These changes may be mainly a consequence of microstructural damage to white matter tracts and reductions in cerebral blood flow. Studies of adolescents and adults with type 2 diabetes reveal the presence of memory dysfunction as well as psychomotor slowing, and these changes are associated with volumetric changes in the hippocampus and other brain structures, Both degree of insulin resistance and higher HbA1c values are correlated with those findings. Research with diabetic children has identified somewhat different predictors. The strongest risk factor is the diagnosis of diabetes early in life [ndash] as the brain is still developing. While poor metabolic control is linked to the occurrence of brain anomalies in this patient population, recurrent episodes of hypoglycemia may also contribute. Very recent work suggests that the metabolic derangements occurring at around the time of diagnosis may play a major role in inducing brain remodeling [ndash] likely via processes occurring at the blood-brain barrier, and these changes may increase the sensitivity of the brain to subsequent metabolic and vascular events.[br][br]Nothing to Disclose: CMR 2012-06-26T16:15:00 Grand Ballroom AB 2012-06-26T00:00:00 1899-12-30T16:15:00 2505 411 2847 PS8-2 SSS06 Tuesday 427 2012


3120 ENDO12L_PS8-3 PRESIDENTIAL SYMPOSIUM SESSION: CLINICAL - Diabetes [amp] The Brain (3:45 PM - 5:15 PM) Non-Glucose Mechanisms of Cognitive Impairment in Type 2 Diabetes Mark WJ Strachan Western General Hospital, Edinburgh, UK Type 2 diabetes is associated with an increased risk of both Alzheimer[apos]s Dementia (AD) and Vascular Dementia (VaD). These two forms of dementia have traditionally been regarded as distinct neuropathological entities. However, MRI studies have demonstrated that [apos]silent[apos] vascular lesions are extremely common in older people and neuropathological investigations have shown that vascular pathology is so common that few individuals with dementia lack a mixed AD and vascular pattern.[br]There are likely to be multiple mechanisms underpinning the relationship between Type 2 diabetes and dementia. Data from experimental studies in animals and from epidemiological studies in humans suggest a link between insulin resistance and cognitive performance and dementia. Insulin receptors are found in high concentrations within the limbic system of the brain. [beta]-amyloid oligomers, which accumulate in the brain in AD and are intimately linked with neuronal dysfunction, cause a rapid and substantial loss of cell surface insulin receptors in hippocampal neuronal cultures. The development of insulin resistance in the brain can then set up a vicious cycle because resultant activation of glycogen synthase kinase 3 (GSK3) promotes hyperphosphorylation of tau protein and altered processing of Amyloid Precursor Protein which increases A[beta]-oligomer production. Moreover, insulin resistance may reduce the ability of insulin degrading enzyme in the brain to breakdown A[beta]-oligomers.[br]Proving the existence of these mechanisms and pathways in humans though has been more challenging. Insulin concentrations in the CSF do appear lower in people with severe AD and administration of intravenous insulin does enhance cognitive function. Studies with Rosiglitazone have shown no benefit of peripheral insulin sensitisation on cognition in people with AD. Intra-nasal insulin may improve some aspects of memory in healthy individuals and pilot data also shows promise in people with AD.[br]The hypothalmic-pituitary-adrenal axis is activated in people with Type 2 diabetes and there are data linking increased cortisol concentrations with cognitive impairment and symptoms of depression. Moreover, Type 2 diabetes is a por-inflammatory state and several studies have shown links with inflammatory markers (TNF, IL-6 and TNF-alpha) and cognitive function. Whether these are causitive factors or simply markers of insulin resistance remains to be elucidated.[br][br]Sources of Research Support: The Edinburgh Type 2 Diabetes Study is supported by the UK Medical Research Council and Diabetes UK.[br][br]Disclosures: MWJS: Consultant, GlaxoSmithKline; Speaker, Eli Lilly & Company, Novo Nordisk, Sanofi-Aventis. 2012-06-26T16:45:00 Grand Ballroom AB 2012-06-26T00:00:00 1899-12-30T16:45:00 2547 411 2848 PS8-3 SSS06 Tuesday 428 2012


3121 ENDO12L_S59-1 SYMPOSIUM SESSION: CLINICAL - New Insights in the Management of Hypothyroidism (3:45 PM - 5:15 PM) Genetic [amp] Clinical Determinants of Thyroid Hormone Replacement Francesco Saverio Celi NIH, Bethesda, MD Thyroid hormone (TH) action depends on the T3 delivered at the end-organ targets and serum T3 level is a function of secretion from the thyroid, peripheral conversion of T4 into T3 or rT3, and catabolism of TH.[br]Hypothyroidism is characterized by lack of endogenous TH production, and its treatment is aimed to restore physiologic amount of TH, and ultimately of T3, in all the tissue targets.[br]Levothyroxine (L-T4) is the standard therapy for hypothyroidism; the restoration of thyroid function is based on the conversion of exogenous L-T4 into the hormonally active T3. The therapeutic goal is a normal TSH, indicating a state of pituitary euthyroidism, on the assumption that it equates to generalized euthyroidism. On the other hand, some patients complain of hypothyroid symptoms despite having thyroid function tests on target.[br]Dysfunction in type-2 deiodinase (D2), which catalyzes the conversion of T4 into T3 in an autocrine/paracrine pattern, may explain this condition. Two D2 polymorphisms have been studied. The inactivating Thr92Ala has minimal effects on TH circulating levels, but hypothyroid carriers of the Ala92 allele had worse quality of life at baseline and improved response to combination therapy. Pharmacogenomic studies indicated that the Ala92 allele is associated to impaired T3 secretion after TRH-induced acute TSH surge, consistent with a decreased intrathyroidal conversion. The -258 A/G is associated with an increased transcription of the D2 gene and a shift in the T3/fT4 ratio at baseline. It is also associated with a blunted fT4 rise after TRH-induced acute TSH surge, consistent with an increased intrathyroidal conversion. Furthermore, the -258G/Thr92 haplotype is associated to a blunted rise in TSH after TRH injection, suggesting a state of relative hyperconversion in the pituitary. Finally, a recent study bypassed the conversion step by treating hypothyroid patients with pharmaco-equivalent does of liothyronine (L-T3) vs. L-T4. The L-T3 treatment resulted in a significant increase in hepatic TH action and in weight loss without any significant change in mood or quality of life.[br]In conclusion, although experimental data indicate the rationale for a pharmacogenomic approach to TH replacement therapy, and that a state of pituitary euthyroidism is not synonymous of generalized optimal delivery of T3, presently there is no strong clinical evidence indicating the need of such therapy or an optimal index of effective TH replacement beyond serum TSH.[br][br]Sources of Research Support: This work was supported by the Intramural Reseach Program of the National Institute of Diabetes, Digestive, and Kidney Diseases.[br][br]Nothing to Disclose: FSC 2012-06-26T15:45:00 Theater A 2012-06-26T00:00:00 1899-12-30T15:45:00 2462 412 2849 S59-1 T02-S04 Tuesday 430 2012


3122 ENDO12L_S59-2 SYMPOSIUM SESSION: CLINICAL - New Insights in the Management of Hypothyroidism (3:45 PM - 5:15 PM) Impact of Environmental Exposures on Thyroid Function Gregory A Brent VA Greater LA Healthcare, Los Angeles, CA Environmental exposures can influence thyroid function by direct actions on the thyroid as well as by triggering autoimmune thyroid disease. Although it is estimated that 70% of the risk of developing autoimmune thyroid disease is due to genetic susceptibility, environmental agents can trigger the onset and accelerate the pace of progression of thyroid disease. Examples of environmental agents or exposures that influence the thyroid include perchlorate, thiocyanate, polychlorinated biphenols, cigarette smoking, radiation from nuclear fallout, medical radiation, and deficient or excess dietary iodine. The iodine-containing medication, amiodarone, can directly influence thyroid function as well as trigger autoimmune thyroid disease. Interferon alpha, especially when used in the treatment of Hepatitis C, is associated with a high incidence of thyroid disease. Many of these environmental agents can also modify the level of thyroxine replacement required to treat patients with established hypothyroidism. These agents may directly influence thyroid function tests used in the diagnosis of hypothyroidism and to monitor thyroxine therapy. A strategy to identify these agents and monitor their effects on the thyroid will be discussed. The impact of these agents on individuals who are most susceptible to their effects, young children and older adults, will be emphasized and the approach to clinical management.[br][br]Nothing to Disclose: GAB 2012-06-26T16:15:00 Theater A 2012-06-26T00:00:00 1899-12-30T16:15:00 2452 412 2850 S59-2 T02-S04 Tuesday 431 2012


3123 ENDO12L_S59-3 SYMPOSIUM SESSION: CLINICAL - New Insights in the Management of Hypothyroidism (3:45 PM - 5:15 PM) Thyroid Hormone Replacement in Pregnancy Kathryn Graham Schuff Oregon Health Sciences University, Portland, OR Awareness of the changes in thyroid homeostasis in normal pregnancy is critical for appropriate pharmacologic therapy and monitoring. TSH levels are lower in pregnancy with suggested trimester-specific normal ranges have been suggested[sup]1[/sup]:[br]First trimester: 0.1 [ndash] 2.5 mIU/L[br]Second trimester: 0.2 [ndash] 3.0 mIU/L[br]Third trimester: 0.3 [ndash] 3.0 mIU/L[br]Early in pregnancy, large increases in thyroxine binding globulin result in higher total T4 levels. Free T4 levels remain normal and decrease slightly throughout gestation, however various analog Free T4 assays have different performance and do not usually report trimester-specific normal ranges[sup]2[/sup].[br]During pregnancy, daily iodine requirement increases by 50%, which can result in the development of hypothyroidism in areas of iodine deficiency. Concerning is that women of child-bearing age in the US may have marginal dietary iodine intake and many prenatal vitamins fail to meet the WHO recommended daily intake of 250 mcg/day[sup]3[/sup].[br]The increase in TBG results in a significant increase in levothyroxine requirements in treated hypothyroid pregnant women. However, the amount of increase varies from 20-80%, dependent on the degree of reserve thyroid function, related to the etiology of hypothyroidism and prepregnancy TSH levels. Because the increase in TBG occurs early in pregnancy (4[sup]th[/sup] [ndash] 6[sup]th[/sup] week), it is recommended that women increase their dose by about 30% (2 pills a week) immediately on discovering they are pregnant and TSH monitoring every 4 weeks[sup]4[/sup] through about the 20[sup]th[/sup] week of gestation, and then once again before delivery.[br]Screening for thyroid dysfunction is controversial. Proponents of screening cite the impact of hypothyroidism on intellectual function of the offspring; opponents call for randomized controlled trials to demonstrate the efficacy of screening procedures and treatment. Recently reported results from the Controlled Antenatal Thyroid Screening Study (CATS) trial did not demonstrate a benefit from antenatal screening at 12 weeks and levothyroxine treatment on cognitive function at 3 years of age[sup]5[/sup]. An NIH-funded trial through the Maternal Fetal Medicine network of women screened and treated by 20 weeks gestation with offspring assessed at 5 years of age is ongoing[sup]6[/sup].[br]Although requiring confirmation as well as evaluation in an iodine sufficient population, levothyroxine treatment in euthyroid women with TPO antibodies demonstrated a dramatic reduction in spontaneous miscarriage and preterm delivery[sup]7[/sup].[br][br](1) Stagnoaro-Green A, Abalovich M, Alexander E, Azizi F, Mestman J, Negro R, et al. 2011 Guidelines of the American Thyroid Association for the Diagnosis and Management of Thyroid Disease During Pregnancy and Postpartum. Thyroid 21:1081-1125. (2) Roti E, Gardini E, Minelli R, Bianconi L, Flisi M 1991 Thyroid function evaluation by different commercially available free thyroid hormone measurement kits in term pregnant women and their newborns. J Endocrinol Invest 14:1-9. (3) Leung AM, Pearce EN, Braverman LE 2009 Iodine content of prenatal multivitamins in the United States. N Engl J Med 360:939-40. (4) Yassa L, Marqusee E, Fawcett R, Alexander EK 2010 Thyroid hormone early adjustment in pregnancy (the THERAPY) trial. J Clin Endocrinol Metab 95:3234-3241. (5) Lazarus JH, Bestwick JP, Channon S, Paradice R, Maina A, Rees R, et al. 2012 Antenatal Thyroid Screening and Childhood Cognitive Function. N Engl J Med 366:493-501. (6) Thyroid Therapy for Mild Thyroid Deficiency in Pregnancy: ClinicalTrials.gov number NCT00388297. (7) Negro R, Formoso G, Mangieri T, Pezzarossa A, Dazzi D, Hassan H 2006 Levothyroxine treatment in euthyroid pregnant women with autoimmune thyroid disease: effects on obstetrical complications. J Clin Endocrinol Metab 91:2587-2591.[br][br]Nothing to Disclose: KGS 2012-06-26T16:45:00 Theater A 2012-06-26T00:00:00 1899-12-30T16:45:00 2512 412 2851 S59-3 T02-S04 Tuesday 432 2012


3124 ENDO12L_S60-1 SYMPOSIUM SESSION: BASIC - Novel Neural Mechanisms Regulating Feeding [amp] Peripheral Metabolism (3:45 PM - 5:15 PM) Melanocortin Regulations of Hepatic Cholesterol Biosynthesis Matthias H Tschoep University of Cincinnati , Cincinnati, OH Disclosure Incomplete: MHT 2012-06-26T15:45:00 Room 342 ABDE 2012-06-26T00:00:00 1899-12-30T15:45:00 6051 413 2852 S60-1 T04-S06 Tuesday 434 2012


3125 ENDO12L_S60-2 SYMPOSIUM SESSION: BASIC - Novel Neural Mechanisms Regulating Feeding [amp] Peripheral Metabolism (3:45 PM - 5:15 PM) Testing Endocrine [amp] Molecular Mechanisms of Bariatric Surgery Darleen A Sandoval University of Cincinnati, Cincinnati, OH Our current behavioral and pharmacological interventions to treat obesity have limited and varied results. Bariatric surgeries are currently the most effective therapy for reliable and sustained weight-loss with the added benefit of reduced rates of comorbidities including diabetes. The accumulating evidence suggests that manipulation of the GI tract that occurs with these surgeries has key metabolic influences that go beyond simple restriction of meal size or nutrient absorption with long-lasting effects on energy and nutrient homeostasis. Determining the mechanisms of these surgeries underscores the ability to drive the development of more effective therapeutic, surgical, or behavioral interventions that can be more widely implemented to treat obesity and diabetes. To this end, our lab has developed rodent models of the vertical sleeve gastrectomy (VSG), a surgery in which 80% of gastric tissue along the greater curvature is removed and the intestine remains intact. After VSG, we demonstrate sustained body weight and fat loss that persists after disappearance of a transient, postsurgical reduction in food intake. VSG produced no obvious weight-loss-independent changes in leptin levels or gene expression of hypothalamic leptin targets and melanocortin 4 receptors are not necessary for the beneficial effects of the surgery on energy and glucose homeostasis. However, there are improvements in both postprandial lipid and glucose handling and macronutrient preference that go beyond the effects of weight-loss. The changes in lipid handling are liver-independent whereas the acute changes in glucose handling are specific to the liver. While VSG increases GLP-1 and decreases ghrelin, neither GLP-1 receptors nor the removal of ghrelin is necessary for the general effects of VSG on body weight and glucose tolerance. These data point to the power of combining advanced physiology, modern genetic models and bariatric surgery to reveal key elements of how the GI tract can influence glucose and lipid metabolism.[br][br]Disclosures: DAS: Research Funding, Johnson & Johnson, Novo Nordisk, Pfizer, Inc.; Speaker, Novo Nordisk. 2012-06-26T16:15:00 Room 342 ABDE 2012-06-26T00:00:00 1899-12-30T16:15:00 2470 413 2853 S60-2 T04-S06 Tuesday 435 2012


3126 ENDO12L_S60-3 SYMPOSIUM SESSION: BASIC - Novel Neural Mechanisms Regulating Feeding [amp] Peripheral Metabolism (3:45 PM - 5:15 PM) Melanocortin-Independent Signaling by AgRP Neurons Modulates Feeding Richard D Palmiter University of Washington, Seattle, WA Neurons in the arcuate region of the hypothalamus that express neuropeptide Y, agouti-related protein (AgRP) and GABA (referred to here as AgRP neurons) are known to counteract the anorexigenic actions of neighboring neurons that express proopiomelanocortin (POMC). However, artificial stimulation of AgRP neurons stimulates robust feeding, whereas their inhibition inhibits feeding and their ablation in adult mice leads to starvation and these effects of AgRP neuron manipulation on feeding still occur in mice with constitutive blockade of POMC signaling. The starvation phenotype following ablation of AgRP neurons is attributable to loss to GABA signaling in the parabrachial nucleus (PBN), which results in unopposed activation of the nucleus. Recent experiments demonstrate that artificial activation of PBN neurons also leads to anorexia, whereas inhibiting these neurons or compromising their glutamatergic output stimulates feeding. Thus, the activity of PBN neurons bidirectionally influences feeding behavior. Current experiments are directed towards identifying the specific neuronal population within the PBN responsible for modulating feeding behavior and determining the critical brain regions affected by those neurons.[br][br]Sources of Research Support: National Institutes of Health and Klarman Family Foundation.[br][br]Nothing to Disclose: RDP 2012-06-26T16:45:00 Room 342 ABDE 2012-06-26T00:00:00 1899-12-30T16:45:00 2441 413 2854 S60-3 T04-S06 Tuesday 436 2012


3127 ENDO12L_S61-1 SYMPOSIUM SESSION: BASIC - Post-Translational Modifications [amp] Nuclear Receptor Action (3:45 PM - 5:15 PM) The Role of Cell Signaling in Steroid Receptor Action Nancy Lynn Weigel Baylor College of Medicine, Houston, TX Steroid receptors, including the progesterone receptor (PR) and androgen receptor (AR), are phosphoproteins as are most of their coregulators. We have employed multiple approaches to elucidate the role of cell signaling pathways in steroid receptor action including identification of phosphorylation sites in the progesterone receptor (PR) and one of its coactivators, SRC-1, modulation of cell signaling pathways and assessing the resulting changes in receptor action, and identification of signaling molecules whose expression or activity is altered upon receptor activation. We have generated breast cancer cell lines expressing PR with single or multiple alanine substitutions for phosphorylation sites and have found that changes in activity are target gene specific. To assess the role of phosphorylation in vivo, we have created a mouse with an alanine substitution for serine 191 (corresponds to Ser190 in human). Most physiological functions measured to date appear normal although there is a modest reduction in fertility. To examine gene expression in the mammary gland, we have developed three dimensional cultures of primary mammary epithelial cells, which retain PR expression. PR in the 191/191 mouse shows a reduced capacity to induce RankL, a key target of PR in the development and function of the mammary gland. Most of the phosphorylation sites identified in the receptors and their coactivators contain Ser/Thr-Pro motifs suggesting that their phosphorylation is regulated by cyclin dependent kinases (Cdks) or mitogen activated kinases (MAPK). Cyclin A interacts with PR and overexpression enhances PR activity. Both cyclin A partners, Cdk1 and Cdk2, play a role in regulating PR activity. Cdks regulate SRC-1 phosphorylation and activity as well. One of the SRC-1 sites is required for optimal coactivation of multiple receptors. Inhibition of MEK and the downstream p42/p44 MAPK using U0126 has minimal effect on PR activity in serum-containing medium, although it strongly inhibits activity in serum free medium. In contrast, AR activity (both full length and a constitutively active splice variant) is sensitive to inhibition of either the CDK or the p42/p44 MAPK pathway. Treatment with U0126 reduces AR transcriptional activity in a target gene specific manner. U0126 reduces recruitment of AR to chromatin and inhibited hormone induced histone acetylation. Thus, cell signaling regulates receptor activity in a receptor specific and a target gene specific manner.[br][br]Nothing to Disclose: NLW 2012-06-26T15:45:00 Room 361 ABDE 2012-06-26T00:00:00 1899-12-30T15:45:00 2504 414 2855 S61-1 T08-S03 Tuesday 438 2012


3128 ENDO12L_S61-2 SYMPOSIUM SESSION: BASIC - Post-Translational Modifications [amp] Nuclear Receptor Action (3:45 PM - 5:15 PM) Modulation of Steroid Receptor Function by SUMOylation Jorma Juhani Palvimo University of Eastern Finland, Kuopio, Kuopio, Finland In addition to hormonal ligands, the activities of steroid receptors are regulated by post-translational modifications (PTMs) which enable other signaling pathways to crosstalk with steroid signaling. SUMOylation, conjugation of small ubiquitin-related modifier (SUMO) protein 1, 2 or 3 to specific lysine residues of target proteins, is a bulky PTM that regulates several important physiological processes. SUMOylations target conserved lysine residues in steroid receptors, including androgen receptor (AR) and glucocorticoid receptor (GR), which modulates their transcriptional activity as assessed by reporter gene assays. Accordingly, SUMOylation pathway components, UBC9 (E2 conjugase), PIAS1 and -3 proteins (E3 ligases) and SENP1 and -2 (SUMO-deconjugases) act as coregulators of AR and GR signaling. To address how SUMOylation influences the activity of endogenous AR and GR in normal chromatin environments, we have compared AR and GR target gene expression and chromatin binding on a genome-wide scale in stable and isogenic cell lines expressing wild-type or SUMOylation-defective receptors. Moreover, we have investigated the effect of SUMO ligase PIAS1 depletion on AR-regulated gene programs in breast and prostate cancer cells. Since cell stress is intimately linked to SUMOylation, enhanced protein modification by SUMO-2 in particular, we have also examined how various types of cell stress, including hyperthermia and reactive oxygen species, influence the SUMOylation and intranuclear compartmentalization of the AR and the GR. Results from these studies, implying that SUMOylations modulate the activity and the chromatin binding of AR and GR in a target gene/gene program selective fashion and the responsiveness of the receptors to cell stress, will be discussed.[br][br]Nothing to Disclose: JJP 2012-06-26T16:15:00 Room 361 ABDE 2012-06-26T00:00:00 1899-12-30T16:15:00 2479 414 2856 S61-2 T08-S03 Tuesday 439 2012


3129 ENDO12L_S61-3 SYMPOSIUM SESSION: BASIC - Post-Translational Modifications [amp] Nuclear Receptor Action (3:45 PM - 5:15 PM) Class I HDACs as Coactivators of GR-Mediated Transcription Catharine Lynn Smith University of Arizona, Tucson, AZ Lysine deacetylases (KDACs or HDACs) have been traditionally thought of as corepressors of transcription. However, they are abundant at active genes and their inhibition results in repression of gene expression as well as the predicted activation. Previously, we showed that transcription from the mouse mammary tumor virus promoter is rapidly repressed upon pharmacological inhibition of KDACs in a manner independent of changes in chromatin remodeling or histone acetylation, suggesting that KDACs target non-histone proteins at this promoter. We have recently reported that KDAC activity facilitates transcriptional initiation at the MMTV promoter through effects on reinitiation from an assembled initiation compex rather than on de novo initiation. In our current studies we have measured the impact of the KDAC inhibitor (KDACi) valproic acid (VPA) on the GR-regulated transcriptome by microarray in a hepatoma cell line. This drug has a profound impact on GR-regulated transcription in several ways, the most predominant of which is that it impairs GR action at 40-50% of glucocorticoid-activated genes. A structurally distinct KDACi, apicidin, which would be unlikely to have similar off-target effects as VPA, was also able to impair activation of these genes. Depletion of KDAC1 expression through siRNA can fully or partially mimic the effects of VPA on a subset of these genes, while depletion of KDAC2 or KDAC3 have no effect on the ability of GR to transactivate its targets. The mechanism by which GR transactivation is impaired is transcriptional in nature and not due to GR processing defects. We hypothesize that KDAC activity is required for GR-dependent assembly of productive transcription complexes, perhaps by removing inhibitory acetylations. ChIP assays are ongoing to measure the effects of KDAC inhibition or depletion on transcription complexes at GR target genes. In conclusion, we provide compelling evidence that KDAC1 cooperates with GR to activate transcription of a significant subset of GR target genes revealing a novel, unrecognized role for KDAC activity in GR transactivation. In addition, our work suggests that KDACi have endocrine-disrupting activities.[br][br]Nothing to Disclose: CLS 2012-06-26T16:45:00 Room 361 ABDE 2012-06-26T00:00:00 1899-12-30T16:45:00 2458 414 2857 S61-3 T08-S03 Tuesday 440 2012


3130 ENDO12L_S62-1 SYMPOSIUM SESSION: CLINICAL - Transition of Care in Adolescents with Chronic Disease (3:45 PM - 5:15 PM) Transitional Care for Turner Syndrome Karen Rhea Rubin CT Children[apos]s Medical Center, Bloomfield, CT Widespread adoption of transition supports as a basic standard of quality care continues to evade us. The Health Care Transition Clinical Report published in the July, 2011 issue of Pediatrics (1) provides us with a transition-planning algorithm to manage a transition process initiated by age 12 years.[br]Our care focus should shift incrementally from being family to adolescent-centered and for girls with Turner Syndrome (TS) from enhancing final height to inducing feminization with gradually increasing doses of estrogen. The use of more physiologic estrogen regimens such as low dose estradiol patch makes it possible for girls with TS to reap the psychological benefits from an age-appropriate timing of puberty without compromising their final adult height.[br]Transition is the ideal time for our patients to be made aware of their health history and of TS[apos]s evolving impact into adulthood and to promote the the development of independent behaviors to prepare for adult self-care. In addition to the previously known susceptibility to autoimmune disease, congenital heart disease, and hypertension, TS individuals have an observed risk increase for several common adult conditions, including type 2 diabetes, coronary artery disease ((CAD), and osteoporosis. Transition is the time to raise awareness of these health risks and assess each patient[apos]s individual risks. Three major therapeutic targets to minimize adult morbidities in TS include obesity, blood pressure, and estrogen. Obesity which excacerbates many of the associated co-morbidities, is increased in TS and progresses during adolescence. Hypertension affects up to 50% of young adults with TS and increases risk for both CAD and aortic rupture. Continuous and sufficient estrogen is the mainstay of bone health and important to sustain vascular and psychosexual health and maintain reproductive organ integrity for potential assisted reproduction.[br]During the final phase of transition, the pediatric endocrinologist should engage the patient in formulating an adolescent-centered adult care plan based on the 2007 TS Clinical Practice Guideline (2) and the needs of the individual patient, closely collaborating with her current and future adult providers to improve the likelihood that she will continue to be carefully monitored in away that optimizes her adult health and longevity.[br][br](1) The Academy of Pediatrics, Pediatrics 2011;128: 182[br](2) Bondy, CA for The Turner Syndrome Consensus Study Group, JCEM 2007;92:10[br][br]Nothing to Disclose: KRR 2012-06-26T15:45:00 Theater B 2012-06-26T00:00:00 1899-12-30T15:45:00 2499 415 2858 S62-1 T07-S05 Tuesday 442 2012


3131 ENDO12L_S62-2 SYMPOSIUM SESSION: CLINICAL - Transition of Care in Adolescents with Chronic Disease (3:45 PM - 5:15 PM) Falling through the Health Care Cracks: Teens with Type 1 Diabetes Transitioning to Adult Care Michael Harris Oregon Health [amp] Science University, Portland, OR The challenges faced by young people with diabetes transitioning from pediatric to adult health care services are numerous and often daunting (Harris et al. 2011). These challenges include developmental milestones of emerging adult hood (Arnett, 2000), differences in pediatric and adult treatment teams, and institutional and health care systems. Although recognition of the issues and problems of transition has increased during the past decade, meaningful systematic changes have not occurred. Our data indicates that the majority of teens with diabetes have not discussed the need for transition from pediatric to adult diabetes care with their families nor have they discussed the transition process with their pediatric providers (Raymond et al. 2011). In addition, almost all youth and parents reported they did not have a transition plan. Interestingly, approximately 3/4 of youth and 2/3 of parents reported they were neither anxious nor worried about the transition process. However, this implies that in a clinic of 1000 patients in the age of transition, [sim]250 patients and [sim]330 parents would be anxious about the transition process. This finding is consistent with our findings demonstrating that more than 25% of previously transitioned emerging adults reported feeling worried when they transitioned to adult care (Duke et al. 2011). Parents and youth reported approximately 17-18 years of age as the ideal time to address transition-related events. Just under 17 years of age was reported as the time to begin discussing transition issues in clinic, and 17 years of age as the time when youth should begin seeing their providers alone. Almost 18 years of age was identified by youth and parents to be the ideal time to transition from a pediatric diabetes provider to an adult diabetes provider. Responses indicate youth and parents believe the discussion of transition and the actual move to adult care should occur over approximately one year. Based on the data presented, recommendations are provided that are specific to pediatric and adult health care teams, patients and families, and health care systems. Suggestions are presented for future research. Finally, practical yet innovative recommendations for pediatric or adult health care providers are proposed (Weissberg-Benchell et al. 2007).[br][br](1) Arnett JJ. Am Psychologist 2000;55:469-480. (2) Harris MA et al. Am J Lifestyle Med 2011;5(1):85-91. (3) Raymond JK et al. [abstract] Diabetes 2011;71:816P. (4) Duke DC et al. [abstract] Diabetes 2011;71:816P. (5) Weissberg-Benchell J. Diab Care 2007;30(10):2441-2446.[br][br]Sources of Research Support: American Diabetes Association Junior Faculty Award; ADA 1-12-JF-46 awarded to Dr. Jennifer K. Raymond; Mentor MAH.[br][br]Nothing to Disclose: MH 2012-06-26T16:15:00 Theater B 2012-06-26T00:00:00 1899-12-30T16:15:00 2403 415 2859 S62-2 T07-S05 Tuesday 443 2012


3132 ENDO12L_S62-3 SYMPOSIUM SESSION: CLINICAL - Transition of Care in Adolescents with Chronic Disease (3:45 PM - 5:15 PM) Transitional Care in Adolescents with Differences of Sex Development (DSD) Sven Olaf Hiort University of Luebeck, Luebeck, Germany DSD comprise a heterogeneous group of heritable abnormalities of sexual determination and differentiation, formerly termed [bdquo]intersexuality[ldquo]. This includes chromosomal as well as monogenic disorders, which inhibit primarily genetic or endocrine pathways of normal sex development. Also, DSD may be part of a complex syndrome of the affected individual. Therefore, initial presentation to medical professionals may be highly variable and may occur at birth, during childhood, but also later during adolescence or even young adulthood. Furthermore, despite recent enormous advances in the understanding of the molecular mechanisms of sex development, the medical decisions made in patients are mostly lacking evidence-based principles and are carried on case by case evaluations. Very critical discussions have focused on approaches on gender assignment and treatment of DSD. Any decision should be based on a correct diagnosis and possible prediction of development during puberty and adulthood of the affected individual. Any decisions on treatment either surgical or pharmacological should be made with an informed consent of the patient. At the time of transition, puberty plays a pivotal role not only because of the physical changes induced by endogenous or supplementary steroids, but also as a time for sexual orientation and promotion of gender identity. Therefore, many aspects from psychosexual guidance to medical treatment including hormone supplementation and surgical procedures have to be considered. The patient should be always be fully aware of the underlying condition and should be able to reflect any therapeutically valid options. This can only be managed by a multidisciplinary team involving many subspecialties from pediatrics, surgical and gynaecological specialities, but also pathology, laboratory medicine, and especially trained psychologists and/or psychiatrists. Most of all, experienced professionalism is needed to deal with these patients, considering the ethical discussions currently under review.[br][br]Sources of Research Support: EuroDSD, funded under the 7th framework programme of the European Commission (201444).[br][br]Nothing to Disclose: SOH 2012-06-26T16:45:00 Theater B 2012-06-26T00:00:00 1899-12-30T16:45:00 2402 415 2860 S62-3 T07-S05 Tuesday 444 2012


3133 ENDO12L_S63-1 SYMPOSIUM SESSION: TRANSLATIONAL - Where Is Hormone Replacement Therapy Ten Years After the WHI? (3:45 PM - 5:15 PM) Current Positions on HRT: An International Perspective Mary Ann Lumsden University of Glasgow, Glasglow, UK Menopausal symptoms have a significant adverse impact on the quality of life of 25% of women going through menopause and may continue beyond 10 years in 20% of these. HRT is the most effective method for treating them with improvement in QoL. It is associated with uncommon, but much publicised, side effects that have distorted judgement on the risks and benefits. The results of WHI in particular have caused the prescription of HRT to decrease by about 50% in the West and also Australia. Licensing is informed by the best available evidence and though flawed, WHI makes a major contribution to the database. Some of the findings are supported by the large observational studies such as Million Women Study, but these data must be reviewed with caution because of bias. It is undoubtedly an effective treatment for osteoporosis but is now recommended for use as a 2[sup]nd[/sup] line treatment only as benefit is lost when treatment is stopped.[br]WHI supported much previous research that suggested that combined HRT is associated with an increase in the risk of breast cancer, which in absolute numbers is small but when expressed as a relative risk looks substantial and alarming. Estrogen-only therapy is associated with decreased risk, even after use is discontinued. The incidence of breast cancer also appears to have fallen in many countries in line with the decrease in HRT use although here are other reasons why this may have occurred such as an increase in mammographic screening but it is a plausible concept. Obesity and alcohol also increase breast cancer risk, but publicity about this is little/ absent in most countries.[br]Original publications from WHI identified an increase in heart disease risk but reanalysis suggests that this is likely to be age related with younger women experiencing no change. The incidence of heart disease at this age is too low to demonstrate benefit conclusively but many studies assessing risk factors support this concept.[br]HRT use has stabilised in most countries and is used at the lowest dose for the shortest time. Many women receive transdermal preparations that have minimal effect on coagulation although there is no evidence that the impact on the breast differs. Natural progesterone may have less impact but no RCT data are available to compare progestagens.[br]More trials are needed to determine the new regimens, their risk and benefit so that women are not denied treatment for their incapacitating symptoms.[br][br]Nothing to Disclose: MAL 2012-06-26T15:45:00 Room 310 2012-06-26T00:00:00 1899-12-30T15:45:00 2492 416 2861 S63-1 T05-S01 Tuesday 446 2012


3134 ENDO12L_S63-2 SYMPOSIUM SESSION: TRANSLATIONAL - Where Is Hormone Replacement Therapy Ten Years After the WHI? (3:45 PM - 5:15 PM) Effect of Estrogens [amp] Progestins on Breast Cancer: Addressing the Controversies Richard J Santen University of Virginia Health System, Charlottesville, VA The Women[apos]s Health Initiative (WHI) originally reported in 2002 that heart disease and breast cancer risks from menopausal hormone therapy (MHT) outweighed the benefits, resulting in a decline in worldwide use by up to 80%. Critical re-examination of the WHI data noted that the average participant age was 63, that few women had symptoms, and that the WHI data might not apply to symptomatic women shortly after menopause. Since that time, additional data suggest that these younger women do not experience an increase in cardiovascular disease and a reduction occurs with estrogen alone. With respect to breast cancer, estrogen alone reduces the risk in all women and does not increase risk in those just entering menopause. The excess risk of veno-thrombotic disease and stroke are age related and the excess risk in younger women is minimal. Based on this new information, the pendulum has swung and the benefits of menopausal hormone therapy outweigh the risks in younger women just entering menopause. Before initiating therapy, the individual risk of the patient for heart disease and breast cancer should be assessed and therapy limited to those at low risk.[br]Recent modeling studies estimate that MHT does not initiate [underline]de novo [/underline]breast cancer but promotes pre-existing, subclinical cancers to grow more rapidly and to reach the diagnostic threshold sooner. The newly developed occult tumor growth and computer incidence models involve several evidence based assumptions, including: 7% average prevalence of breast cancer at autopsy, average doubling time of 200 days, and mammographic detection threshold of 1.2cm. Based on these assumptions, it is estimated that [underline]de novo[/underline] cancers require more than 10 years to reach the mammographic diagnostic threshold. These models estimate that only 7% of [underline]de novo [/underline]cancers could have become detectable over the 7 years of the WHI study. Accordingly, 93% of the tumors diagnosed during the WHI were pre-existing and stimulated to grow by estrogen plus progestogen administration. The effect of estrogen alone to reduce breast cancer incidence likely represents a pro-apoptotic effect on pre-existing, occult tumors. These considerations, taken together, should serve to reassure younger women with symptoms from menopause to consider use of menopausal hormone therapy.[br][br]Disclosure Incomplete: RJS 2012-06-26T16:15:00 Room 310 2012-06-26T00:00:00 1899-12-30T16:15:00 2451 416 2862 S63-2 T05-S01 Tuesday 447 2012


3135 ENDO12L_S63-3 SYMPOSIUM SESSION: TRANSLATIONAL - Where Is Hormone Replacement Therapy Ten Years After the WHI? (3:45 PM - 5:15 PM) Current Understanding of the Cardiovascular Risks of Hormone Replacement Therapy Howard N Hodis University of Southern Calilfornia, Los Angeles, CA Over the past decade two informative events relevant to women[apos]s health have occurred for the primary prevention of coronary heart disease (CHD). The first concerns hormone replacement therapy (HRT) where data have come full circle from presumed harm to consistency with observational data that HRT initiation in close proximity to menopause significantly reduces CHD by 32% (RR=0.68, 95% CI, 0.48-0.96) and total mortality by 39% (RR=0.61; 95% CI, 0.39-0.95). The other concerns sex-specific efficacy of CHD primary prevention therapies where lipid-lowering and aspirin therapy have not been conclusively shown to significantly reduce CHD in women and more importantly where there is lack of evidence that either therapy reduces total mortality in women. Cumulated data support the [quot]timing hypothesis[quot] for maximal reduction of CHD and total mortality and minimization of risks with HRT initiation before 60 years of age and/or within 10 years of menopause and continued for 6 years or more. Additionally, there is a substantial increase in quality-adjusted life-years over a 5-30 year period in women who initiate HRT in close proximity to menopause supporting HRT as a highly cost-effective strategy for improving quality-adjusted life. Although primary prevention therapies and HRT contrast in their efficacy to significantly reduce CHD and especially total mortality in postmenopausal women, the magnitude and types of risks associated with HRT are similar to those associated with other medications commonly used in women[apos]s health specifically and in clinical medicine generally. The cumulated data highlight the importance of studying the HRT cardioprotective hypothesis in women representative of those from whom the hypothesis was generated, namely in young postmenopausal women in close proximity to menopause.[br][br](1) Hodis HN et al., Climacteric 2012; In Press. (2) Hodis HN et al., Brain Research 2011;1379:244. (3) Hodis HN et al., Menopause 2007;14:944. (4) Hodis HN et al., N Engl J Med 2003;349:535. (5) Hodis HN et al., Ann Intern Med 2001;135:939.[br][br]Sources of Research Support: National Institutes of Health, National Institute on Aging, R01AG-024154.[br][br]Nothing to Disclose: HNH 2012-06-26T16:45:00 Room 310 2012-06-26T00:00:00 1899-12-30T16:45:00 2502 416 2863 S63-3 T05-S01 Tuesday 448 2012


3136 ENDO12L_M47A MEET-THE-PROFESSOR: CLINICAL - Raising HDL: When [amp] How? (Aim High) (5:30 PM - 6:15 PM) Raising HDL: When & How? (Aim High) Ira Jay Goldberg Columbia University, New York, NY Session supported by: Merck & Co., Inc. 2012-06-25T17:30:00 Room 342 ABDE 2012-06-25T00:00:00 1899-12-30T17:30:00 6290 342 2653 M47 L2A Monday 2012


3137 ENDO12L_CMF1-1A CASE MANAGEMENT FORUM: CLINICAL - Pitfalls [amp] Lab-omas in Endocrine Testing [amp] Interpretation (11:15 AM - 12:00 PM) Pitfalls [amp] Lab-omas in Endocrine Testing [amp] Interpretation Robert Alan Vigersky Walter Reed National Military Medical Center, Washington, DC Nothing to Disclose: RAV 2012-06-23T11:15:00 Room 370 2012-06-23T00:00:00 1899-12-30T11:15:00 6175 29 50 CMF1-1 CMF-M1A Saturday 3 2012


3138 ENDO12L_CMF1-2A CASE MANAGEMENT FORUM: CLINICAL - Pitfalls [amp] Lab-omas in Endocrine Testing [amp] Interpretation (11:15 AM - 12:00 PM) Pitfalls [amp] Lab-omas in Endocrine Testing [amp] Interpretation D Robert Dufour VA Medical Center, Laurel, MD Disclosures: DRD: Speaker, Ortho Clinical Diagnostics. 2012-06-23T11:15:00 Room 370 2012-06-23T00:00:00 1899-12-30T11:15:00 6176 29 51 CMF1-2 CMF-M1A Saturday 4 2012


3139 ENDO12L_M1A MEET-THE-PROFESSOR: CLINICAL - Approach to Management of Prediabetic States (Including PCOS) in a Multi-Ethnic Population (11:15 AM - 12:00 PM) Approach to Management of Prediabetic States (Including PCOS) in a Multi-Ethnic Population Samuel Dagogo-Jack University of Tennessee - Memphis, Memphis, TN Disclosures: SD-J: Principal Investigator, Astra Zeneca, Novo Nordisk; Consultant, Merck & Co. 2012-06-23T11:15:00 Room 362 2012-06-23T00:00:00 1899-12-30T11:15:00 6138 30 52 M1 D1A Saturday 5 2012


3140 ENDO12L_M2A MEET-THE-PROFESSOR: CLINICAL - Graves[apos] Ophthalmopathy (11:15 AM - 12:00 PM) Graves[apos] Ophthalmopathy Henry Burdette Burch Walter Reed National Military Medical Center, Potomac, MD Disclosure Incomplete: HBB 2012-06-23T11:15:00 Room 332 2012-06-23T00:00:00 1899-12-30T11:15:00 6158 31 53 M2 TH1A Saturday 6 2012


3141 ENDO12L_M3A MEET-THE-PROFESSOR: CLINICAL - Fatty Liver Disease in Childhood Obesity (11:15 AM - 12:00 PM) Liver Disease in Childhood Obesity Sonia Caprio Yale University, New Haven, CT Nothing to Disclose: SC 2012-06-23T11:15:00 Room 361 ABDE 2012-06-23T00:00:00 1899-12-30T11:15:00 6183 32 54 M3 PED3A Saturday 7 2012


3142 ENDO12L_M5A MEET-THE-PROFESSOR: CLINICAL - Atypical Subtrochanteric [amp] Femoral Shaft Fractures: Role of Bisphosphonates (11:15 AM - 12:00 PM) Atypical Subtrochanteric [amp] Femoral Shaft Fractures: Role of Bisphosphonates Elizabeth Shane Columbia University College of Physicians [amp] Surgeons, New York, NY Nothing to Disclose: ES 2012-06-23T11:15:00 Room 351 2012-06-23T00:00:00 1899-12-30T11:15:00 6243 33 55 M5 B5A Saturday 8 2012


3143 ENDO12L_CMF2-1A CASE MANAGEMENT FORUM: CLINICAL - Cushing[apos]s Disease: Dialogue between an Endocrinologist [amp] a Neurosurgeon (5:30 PM - 6:15 PM) Cushing[apos]s Disease: Dialogue between a Surgeon [amp] an Endocrinologist Specializing in Pituitary Disease James W Findling Medical College of Wisconsin, Menomonee Falls, WI Nothing to Disclose: JWF 2012-06-23T17:30:00 Theater B 2012-06-23T00:00:00 1899-12-30T17:30:00 6260 103 903 CMF2-1 CMF-A1A Saturday 64 2012


3144 ENDO12L_CMF2-2A CASE MANAGEMENT FORUM: CLINICAL - Cushing[apos]s Disease: Dialogue between an Endocrinologist [amp] a Neurosurgeon (5:30 PM - 6:15 PM) Cushing[apos]s Disease: Dialogue between a Surgeon [amp] an Endocrinologist Specializing in Pituitary Disease Brooke Swearingen Massachusetts General Hospital, Boston, MA Disclosure Incomplete: BS 2012-06-23T17:30:00 Theater B 2012-06-23T00:00:00 1899-12-30T17:30:00 6250 103 904 CMF2-2 CMF-A1A Saturday 65 2012


3145 ENDO12L_CMF3-1A CASE MANAGEMENT FORUM: CLINICAL - Diagnosis [amp] Management of Male Hypogonadism in the Older Man (5:30 PM - 6:15 PM) Diagnosis [amp] Management of Male Hypogonadism in the Older Man Frances J Hayes St Vincent[apos]s University Hospital, Dublin, Ireland Disclosure Incomplete: FJH 2012-06-23T17:30:00 Theater A 2012-06-23T00:00:00 1899-12-30T17:30:00 6168 104 905 CMF3-1 CMF-MR1A Saturday 66 2012


3146 ENDO12L_CMF3-2A CASE MANAGEMENT FORUM: CLINICAL - Diagnosis [amp] Management of Male Hypogonadism in the Older Man (5:30 PM - 6:15 PM) Diagnosis [amp] Management of Male Hypogonadism in the Older Man Alvin M Matsumoto VA Puget Sound Health Care System, Seattle, WA Disclosures: AMM: Principal Investigator, Abbott Laboratories, GlaxoSmithKline, Ascend; Consultant, Novartis Pharmaceuticals. 2012-06-23T17:30:00 Theater A 2012-06-23T00:00:00 1899-12-30T17:30:00 6172 104 906 CMF3-2 CMF-MR1A Saturday 67 2012


3147 ENDO12L_M6A MEET-THE-PROFESSOR: CLINICAL - Challenging Multiple Endocrine Neoplasia Cases (5:30 PM - 6:15 PM) Challenging Multiple Endocrine Neoplasia Cases Robert F Gagel University of Texas MD Anderson Cancer Center, Houston, TX Nothing to Disclose: RFG 2012-06-23T17:30:00 Room 351 2012-06-23T00:00:00 1899-12-30T17:30:00 6210 105 907 M6 M1A Saturday 68 2012


3148 ENDO12L_M7A MEET-THE-PROFESSOR: CLINICAL - Chronic Kidney Disease [amp] Bone (5:30 PM - 6:15 PM) Chronic Kidney Disease [amp] Bone Mark Stuart Cooper University of Birmingham, Birmingham, UK Nothing to Disclose: MSC 2012-06-23T17:30:00 Room 371 2012-06-23T00:00:00 1899-12-30T17:30:00 6144 106 908 M7 B7A Saturday 69 2012


3208 ENDO12L_M50A MEET-THE-PROFESSOR: CLINICAL - Biochemical Markers of Bone Turnover in Osteoporosis (1:00 PM - 1:45 PM) Biochemical Markers of Bone Turnover in Osteoporosis Roberto Civitelli Washington University -St Louis, Saint Louis, MO Disclosures: RC: Stockholder, Eli Lilly & Company, Amgen, Merck & Co.; Principal Investigator, Pfizer, Inc. 2012-06-26T13:00:00 Room 370 2012-06-26T00:00:00 1899-12-30T13:00:00 6174 391 2786 M50 B9A Tuesday 370 2012


3209 ENDO12L_M51A MEET-THE-PROFESSOR: CLINICAL - Congenital Adrenal Hyperplasia from Adolescent to Adult (1:00 PM - 1:45 PM) Congenital Adrenal Hyperplasia from Adolescent to Adult Richard Joseph Auchus University of Michigan, Ann Arbor, MI Disclosures: RJA: Investigator, Jansen Pharmaceuticals. 2012-06-26T13:00:00 Room 372 2012-06-26T00:00:00 1899-12-30T13:00:00 6136 392 2787 M51 A1A Tuesday 371 2012


3210 ENDO12L_M52A MEET-THE-PROFESSOR: CLINICAL - Diagnosis [amp] Treatment of Type 2 Diabetes in Youth (1:00 PM - 1:45 PM) Diagnosis [amp] Treatment of Type 2 Diabetes in Youth Francine Ratner Kaufman University Childrens Medical Group, Los Angeles, CA Session supported by: Lilly USA, LLC[br][br]Disclosures: FRK: Chief Scientific Officer, Medtronic Minimed. 2012-06-26T13:00:00 Room 332 2012-06-26T00:00:00 1899-12-30T13:00:00 6163 393 2788 M52 D8A Tuesday 372 2012


3211 ENDO12L_M53A MEET-THE-PROFESSOR: CLINICAL - Pediatric Type 1 Diabetes (1:00 PM - 1:45 PM) Pediatric Type 1 Diabetes Morey W Haymond Children[apos]s Nutrition Research Center, Houston, TX Session supported by: Lilly USA, LLC[br][br]Disclosure Incomplete: MWH 2012-06-25T13:00:00 Room 320 2012-06-26T00:00:00 1899-12-30T13:00:00 6244 394 2789 M53 D2A Tuesday 373 2012


3212 ENDO12L_M54A MEET-THE-PROFESSOR: CLINICAL - Tumor-Induced Osteomalacia (1:00 PM - 1:45 PM) Tumor-Induced Osteomalacia Suzanne Marie Jan De Beur Johns Hopkins University School of Medicine, Baltimore, MD Disclosure Incomplete: SMJDB 2012-06-25T13:00:00 Room 361 ABDE 2012-06-26T00:00:00 1899-12-30T13:00:00 6140 395 2790 M54 B1A Tuesday 374 2012


3234 ENDO12L1_7001 PRE-CONFERENCE EVENT: Endocrine Trainee Day - Class of 2012 (8:00 AM - 5:30 PM) Endocrine Trainee Day [ndash] Class of 2012 Supported by Merck & Co., Inc. and Novo Nordisk Inc. 2012-06-22T08:00:00 George R. Brown Convention Center 2012-06-22T00:00:00 1899-12-30T08:00:00 7001 1 1 9901a Friday 0 2012


3235 ENDO12L1_7002 PRE-CONFERENCE EVENT: Advanced Hands-On Thyroid Ultrasound Workshop (8:00 AM - 5:30 PM) Advanced Hands-On Thyroid Ultrasound Workshop Registration and fee required 2012-06-22T08:00:00 George R. Brown Convention Center 2012-06-22T00:00:00 1899-12-30T08:00:00 7002 2 2 9901b Friday 0 2012


3236 ENDO12L1_7003 PRE-CONFERENCE EVENT: Hormones and Cancer Forum (9:00 AM - 6:00 PM) Hormones and Cancer Forum Registration and fee required 2012-06-22T09:00:00 George R. Brown Convention Center 2012-06-22T00:00:00 1899-12-30T09:00:00 7003 3 3 9902a Friday 0 2012


3237 ENDO12L1_7004 PRE-CONFERENCE EVENT: Modulators of GPCR Trafficking and Signaling (9:00 AM - 6:00 PM) Modulators of GPCR Trafficking and Signaling Registration and fee required 2012-06-22T09:00:00 George R. Brown Convention Center 2012-06-22T00:00:00 1899-12-30T09:00:00 7004 4 4 9902b Friday 0 2012


3238 ENDO12L1_7005 PRE-CONFERENCE EVENT: Diabetes Diagnosis and Management Workshop (9:30 AM - 5:30 PM) Diabetes Diagnosis and Management Workshop Registration and fee required[br][br]Supported by Amylin Pharmaceuticals, Inc., Bristol- Myers Squibb and AstraZeneca LP, Merck [amp] Co., Inc. And Novo Nordisk Inc. 2012-06-22T09:30:00 George R. Brown Convention Center 2012-06-22T00:00:00 1899-12-30T09:30:00 7005 5 5 9903 Friday 0 2012


3239 ENDO12L1_7006 PRE-CONFERENCE EVENT: How To Secure Promotion and Tenior Workshop (1:00 PM - 6:30 PM) How To Secure Promotion and Tenior Workshop Novo Nordisk, Inc. 2012-06-22T13:00:00 George R. Brown Convention Center 2012-06-22T00:00:00 1899-12-30T13:00:00 7006 6 6 9904 Friday 0 2012


3159 ENDO12L_CMF5-2A CASE MANAGEMENT FORUM: CLINICAL - Primary Hyperaldosteronism (1:00 PM - 1:45 PM) Interaction of Aldosterone [amp] Salt on Cardiovascular Remodeling Michael Stowasser University of Queensland School of Medicine, Brisbane, Australia It is now well recognized that aldosterone (aldo) excess in primary aldosteronism (PA) results in cardiovascular (CV) and renal damage and increased CV morbidity independent of its effects on BP. Experimental studies have indicated that the combined effects of aldo excess and high-salt ingestion are necessary for induction of target-organ damage. We have studied the role of salt in determining urinary protein (Uprot) excretion and changes in left ventricular (LV) structure and the relationship between aldo and salt appetite in patients with PA. Subjects (n=24) with aldo-producing adenoma (APA) underwent measurement of 24 h Uprot and UNa before and after unilateral adrenalectomy (ADX - follow-up 15[plusmn]12 months SD). Following ADX, mean clinic systolic BP fell (150[plusmn]18 vs 135[plusmn]15mmHg, p=0.0008), despite reduction in antihypertensive medications, and Uprot decreased (211[plusmn]102 vs 106[plusmn]42mg per day, P[lt]0.0001). There was a positive correlation between Uprot and UNa both before (r=0.55, p=0.006) and after (r=0.51, p=0.011) ADX and changes in UNa independently predicted Uprot reduction (p=0.019). In a case-control study of 21 patients with PA and 21 with essential hypertension (EH) matched for age, gender, duration of hypertension and 24h systolic and diastolic BP, PA patients had significantly greater mean LV wall thicknesses and mass. UNa positively correlated with, and was an independent predictor of, LV wall thicknesses and LV mass in PA but not EH patients. We also compared pre- and post-ADX UNa, in 93 patients with APA collected on the last day (day 4) of fludrocortisone suppression testing (FST), and, in 35 patients, collected out of hospital. Day 4 UNa decreased from 328[plusmn]79 mmol/day pre-operatively to 272[plusmn]69 mmol/day (p[lt]0.0001) postoperatively despite equal salt supplementation. Out of hospital UNa fell from 172[plusmn]57 to 147[plusmn]56 mmol/day (p=0.0183). Interestingly, UNa on day 4 FST fell significantly only in patients whose FST after ADX showed biochemical cure of PA (n=78), but not in those with remaining autonomous aldo production (n=15). The above results indicate that dietary salt modulates aldo-induced renal and cardiac damage in patients with PA. Hence, apart from that afforded by reduction of aldo effects (by ADX or mineralocorticoid receptor blockade), dietary salt restriction should provide additional target-organ protection in patients with PA. This may be easier to achieve in treated patients as aldo reduction appears to reduce salt appetite.[br][br]Sources of Research Support: Irene Hunt Hypertension Research Trust.[br][br]Nothing to Disclose: MS 2012-06-24T13:00:00 Room 362 2012-06-24T00:00:00 1899-12-30T13:00:00 2486 162 1041 CMF5-2 CMF-A2A Sunday 122 2012


3160 ENDO12L_M16A MEET-THE-PROFESSOR: CLINICAL - Adult Turner Syndrome (1:00 PM - 1:45 PM) Adult Turner Syndrome Carolyn Ann Bondy NIH - NICHD, Bethesda, MD Disclosure Incomplete: CAB 2012-06-24T13:00:00 Room 361 ABDE 2012-06-24T00:00:00 1899-12-30T13:00:00 6242 163 1042 M16 FR3A Sunday 123 2012


3161 ENDO12L_M17A MEET-THE-PROFESSOR: CLINICAL - Causes of the Condition [amp] the Epidemic (1:00 PM - 1:45 PM) Childhood Obesity: Causes Tamara S Hannon Riley Hospital for Children, Indianapolis, IN Nothing to Disclose: TSH 2012-06-24T13:00:00 Room 360 2012-06-24T00:00:00 1899-12-30T13:00:00 6191 164 1043 M17 PED1A Sunday 124 2012


3162 ENDO12L_M18A MEET-THE-PROFESSOR: CLINICAL - Hereditary Pheochromocytoma [amp] Paraganglioma (1:00 PM - 1:45 PM) Hereditary Pheochromocytoma [amp] Paraganglioma Karel Pacak PREB, NICHD, Bethesda, MD Disclosure Incomplete: KP 2012-06-24T13:00:00 Room 370 2012-06-24T00:00:00 1899-12-30T13:00:00 6154 165 1044 M18 M2A Sunday 125 2012


3163 ENDO12L_M19A MEET-THE-PROFESSOR: CLINICAL - NASH/NAFLD: How To Treat [amp] Whom To Treat? (1:00 PM - 1:45 PM) NASH/NAFLD: How To Treat [amp] Whom To Treat? Kenneth Cusi University Texas Health Science Center San Antonio, San Antonio, TX Disclosure Incomplete: KC 2012-06-24T13:00:00 Room 352 DEF 2012-06-24T00:00:00 1899-12-30T13:00:00 6152 166 1045 M19 L4A Sunday 126 2012


2766 ENDO12L_L1-1 PLENARY: TRANSLATIONAL - Presidential Plenary Lecture: The National Institute of Child Health [amp] Human Development: 50 Years Old [amp] Looking to the Future (7:45 AM - 9:15 AM) The National Institute of Child Health [amp] Human Development: 50 Years Old [amp] Looking to the Future Alan E Guttmacher National Institute of Child Health [amp] Human Development, Bethesda, MD Session supported by: Eunice Kennedy Shriver National Institute of Child Health [amp] Human Development (NICHD) Grant R13 HD072675[br][br]Disclosure Incomplete: AEG 2012-06-23T07:45:00 Grand Ballroom ABC 2012-06-23T00:00:00 1899-12-30T07:45:00 6119 11 11 L1-1 PL01-1 Saturday 1 2012


2767 ENDO12L_L1-2 PLENARY: TRANSLATIONAL - Presidential Plenary Lecture: Next Generation Human Genetics: A Path to New Biology (7:45 AM - 9:15 AM) Next Generation Human Genetics: A Path to New Biology Joel N Hirschhorn Children[apos]s Hospital, Boston, MA Genome-wide association (GWA) studies have identified thousands of common genetic variants that influence human phenotypes, providing clues to novel and causal biology. For some traits and diseases, human genetic studies have provided clear hypotheses about underlying causal biology, but in many cases, additional genetic and computational work is needed before functional studies can productively be initiated. These follow-up genetic, computational and functional studies will all enable the transition from genetic discovery to biological insight, which remains a main goal of human genetic studies. Next-generation sequencing methods, which enable comprehensive surveys of variation in exons or whole genomes, will further reshape genetic research, and will likely alter clinical practice. Specific examples from ongoing studies of anthropometric traits will be discussed, as will the general principles that have emerged from GWA studies in humans.[br][br]Nothing to Disclose: JNH 2012-06-23T07:45:00 Grand Ballroom ABC 2012-06-23T00:00:00 1899-12-30T07:45:00 2412 12 12 L1-2 PL01-2 Saturday 2 2012


2768 ENDO12L_CMF1-1 CASE MANAGEMENT FORUM: CLINICAL - Pitfalls [amp] Lab-omas in Endocrine Testing [amp] Interpretation (8:30 AM - 9:15 AM) Pitfalls [amp] Lab-omas in Endocrine Testing [amp] Interpretation Robert Alan Vigersky Walter Reed National Military Medical Center, Washington, DC Nothing to Disclose: RAV 2012-06-23T08:30:00 Room 370 2012-06-23T00:00:00 1899-12-30T08:30:00 6175 13 13 CMF1-1 CMF-M1 Saturday 3 2012


2769 ENDO12L_CMF1-2 CASE MANAGEMENT FORUM: CLINICAL - Pitfalls [amp] Lab-omas in Endocrine Testing [amp] Interpretation (8:30 AM - 9:15 AM) Pitfalls [amp] Lab-omas in Endocrine Testing [amp] Interpretation D Robert Dufour VA Medical Center, Laurel, MD Disclosures: DRD: Speaker, Ortho Clinical Diagnostics. 2012-06-23T08:30:00 Room 370 2012-06-23T00:00:00 1899-12-30T08:30:00 6176 13 14 CMF1-2 CMF-M1 Saturday 4 2012


2770 ENDO12L_M1 MEET-THE-PROFESSOR: CLINICAL - Approach to Management of Prediabetic States (Including PCOS) in a Multi-Ethnic Population (8:30 AM - 9:15 AM) Approach to Management of Prediabetic States (Including PCOS) in a Multi-Ethnic Population Samuel Dagogo-Jack University of Tennessee - Memphis, Memphis, TN Disclosures: SD-J: Principal Investigator, Astra Zeneca, Novo Nordisk; Consultant, Merck & Co. 2012-06-23T08:30:00 Room 362 2012-06-23T00:00:00 1899-12-30T08:30:00 6138 14 15 M1 D1 Saturday 5 2012


2771 ENDO12L_M2 MEET-THE-PROFESSOR: CLINICAL - Graves[apos] Ophthalmopathy (8:30 AM - 9:15 AM) Graves[apos] Ophthalmopathy Henry Burdette Burch Walter Reed National Military Medical Center, Potomac, MD Disclosure Incomplete: HBB 2012-06-23T08:30:00 Room 332 2012-06-23T00:00:00 1899-12-30T08:30:00 6158 15 16 M2 TH1 Saturday 6 2012


2772 ENDO12L_M3 MEET-THE-PROFESSOR: CLINICAL - Fatty Liver Disease in Childhood Obesity (8:30 AM - 9:15 AM) Liver Disease in Childhood Obesity Sonia Caprio Yale University, New Haven, CT Nothing to Disclose: SC 2012-06-23T08:30:00 Room 361 ABDE 2012-06-23T00:00:00 1899-12-30T08:30:00 6183 16 17 M3 PED3 Saturday 7 2012


2773 ENDO12L_M5 MEET-THE-PROFESSOR: CLINICAL - Atypical Subtrochanteric [amp] Femoral Shaft Fractures: Role of Bisphosphonates (8:30 AM - 9:15 AM) Atypical Subtrochanteric [amp] Femoral Shaft Fractures: Role of Bisphosphonates Elizabeth Shane Columbia University College of Physicians [amp] Surgeons, New York, NY Nothing to Disclose: ES 2012-06-23T08:30:00 Room 351 2012-06-23T00:00:00 1899-12-30T08:30:00 6243 17 18 M5 B5 Saturday 8 2012


2774 ENDO12L_S1-1 SYMPOSIUM SESSION: TRANSLATIONAL - Animal Models for PCOS (9:30 AM - 11:00 AM) Antecedents of Metabolic Dysfunction in a Nonhuman Primate Model of PCOS David H Abbott University of Wisconsin-National Primate Research Center, Madison, WI Fetal androgen excess induces polycystic ovary syndrome (PCOS)-like reproductive and metabolic traits in female mammals from rodents to primates. Recent studies in female rhesus monkeys demonstrate that maternal glucose intolerance is induced in tandem with androgen excess (1). Pancreatic accommodation of dual androgenic and glycemic gestational excess may contribute to adult metabolic pathophysiology in these prenatally androgenized (PA) female monkeys. To gain insight into potential molecular mechanisms involved in PA monkey pathophysiology, DNA methylation was examined in visceral fat from PA infant and adult females. Pathway and network analyses of differentially methylated genes implicated multiple signaling pathways (2). Transforming growth factor-beta (TGF-beta) signaling was amongst the top two pathways identified by gene methylation analyses in both PA infant and adult monkeys. As a potential gene candidate for PCOS, intron 55 of fibrillin 3, is involved in regulating TGF-beta signaling in women, altered function of this signaling pathway may accompany PCOS pathophysiology in PA female monkeys and in women.[br][br](1) Abbott DH et al., Am J Physiol Endocrinol Metab 2010; 299:E741. (2) Xu N et al., PLoS ONE 6:e27286.[br][br]Sources of Research Support: NIH grants R01-HD029364, K24-HD001346, R01-DK079888, R01-RR013635, P50-HD044405, U01-HD044650, P51-RR000167, P51-RR000169, M01-RR000425.[br][br]Disclosures: DHA: Ad Hoc Consultant, Principal Investigator, Viamet Pharmaceuticals, Inc. 2012-06-23T09:30:00 Room 362 2012-06-23T00:00:00 1899-12-30T09:30:00 2513 18 19 S1-1 T05-S06 Saturday 10 2012


2775 ENDO12L_S1-2 SYMPOSIUM SESSION: TRANSLATIONAL - Animal Models for PCOS (9:30 AM - 11:00 AM) Developmental Origin of PCOS: Androgenic vs Estrogenic Reprogramming Vasantha Padmanabhan University of Michigan, Ann Arbor, MI Polycystic ovary syndrome (PCOS) is the most common reproductive disorder affecting several million women worldwide. Women with PCOS manifest neuroendocrine, ovarian and metabolic defects. A large number of animal models have been developed to understand the etiology of PCOS. Using sheep as a model our studies have found that exposure to excess testosterone during fetal life results in a PCOS phenotype, which includes oligo-/an-ovulation, functional hyperandrogenism, polycystic ovarian morphology, LH excess, neuroendocrine feedback defects, insulin resistance and hypertension. Importantly, postnatal overfeeding was found to amplify the severity of the reproductive phenotype. Because testosterone can be aromatized to estrogen, it is unclear which attributes of these anomalies is the result of reprogramming due to androgenic as opposed to estrogenic actions of testosterone. Comparison of reproductive and metabolic outcomes from prenatal testosterone (aromatizable androgen) treatment with outcomes from prenatal dihydrotestosterone (non-aromatizable androgen) treatment or outcomes from co-treatment with testosterone and flutamide (androgen antagonist) has helped delineate the relative roles of androgens and estrogens in programming the PCOS phenotype. These studies have found that reduced sensitivity to estradiol negative feedback, luteinizing hormone excess, increased pituitary sensitivity to gonadotropin-releasing hormone, and insulin resistance evident in prenatal testosterone-treated females are mediated via androgenic programming. In contrast, disruption in estradiol positive feedback, increased estradiol release, mulitfollicular ovarian phenotype, enhanced follicular persistence, increased ovarian androgen receptor expression, appear to be programmed via estrogenic actions of prenatal testosterone. These findings are likely to help in targeting appropriate intervention strategies aimed towards improving fertility and metabolic outcomes and therefore are of translational relevance.[br][br]Sources of Research Support: NIH P01 HD44232.[br][br]Nothing to Disclose: VP 2012-06-23T10:00:00 Room 362 2012-06-23T00:00:00 1899-12-30T10:00:00 2413 18 20 S1-2 T05-S06 Saturday 11 2012


2776 ENDO12L_S1-3 SYMPOSIUM SESSION: TRANSLATIONAL - Animal Models for PCOS (9:30 AM - 11:00 AM) The Effect of Postnatal Androgen Exposure on Reproductive [amp] Metabolic Phenotypes in Rodents as a Model of PCOS Elisabet Stener-Victorin Institute of Neuroscience [amp] Physiology, Goteborg, Sweden Animal PCOS models are needed when making the transition from scientific concepts to attaining an understanding of a human disease. Recently, two new rat PCOS model that incorporates ovarian [italic]and[/italic] metabolic characteristics of the syndrome was developed [1]. Since most women start to develop their PCOS symptoms during early puberty, at the same time as the androgens are starting to be produced, the continuous dihydrotestosterone (DHT), a nonaromatizable androgen, and letrozole, a non-steroidal inhibitor of P450 aromatase exposure started pre-pubertal in order to study the contribution of androgens on female rats at adult age as a conceivable PCOS model. After continuous exposure to DHT, from prepuberty until adult age, the rats have typical PCO with an increased number of apoptotic follicles [1]. Moreover, the DHT exposed rats develop obesity accompanied by enlarged adipocyte size and insulin resistance, indicating that high levels of androgens induce alterations in body composition and reduced insulin sensitivity [1]. In addition, the DHT exposed rats also develop endothelial dysfunction [2]. The DHT-induced rat PCOS model has recently been applied in mice with development of a similar phenotype [3]. Results from a recent dose-response study with letrozole exposure will be presented. Similarities and dissimilarities with human PCOS as well as advantage and disadvantage of different rodent models will be discussed.[br][br]1. Manner[aring]s L, Cajander S, Holm[auml]ng A, Seleskovic Z, Lystig T, et al. (2007) A new rat model exhibiting both ovarian and metabolic characteristics of polycystic ovary syndrome. Endocrinology 148: 3781-3791. 2. Keller J, Mandala M, Casson P, Osol G (2011) Endothelial dysfunction in a rat model of PCOS: evidence of increased vasoconstrictor prostanoid activity. Endocrinology 152: 4927-4936. 3. van Houten AF, Kramer P, McLuskey A, Karels B, Themmen A, et al. (2012) Reproductive and metabolic phenotype of a mouse model of PCOS. Endocrinology 153: 0000-0000.[br][br]Sources of Research Support: Swedish Medical Research Council (Project No. 2008-72VP-15445-01A and K2012-55X-15276-08-3); Novo Nordisk Foundation; Swedish federal government under the LUA/ALF agreement ALFFGBG-136481.[br][br]Nothing to Disclose: ES-V 2012-06-23T10:30:00 Room 362 2012-06-23T00:00:00 1899-12-30T10:30:00 2390 18 21 S1-3 T05-S06 Saturday 12 2012


2777 ENDO12L_S2-1 SYMPOSIUM SESSION: CLINICAL - Complications of Childhood Obesity (9:30 AM - 11:00 AM) Ethnicity, BMI [amp] Risk of Type 2 Diabetes Fida F Bacha Baylor College of Medicine, Houston, TX The increase in the incidence of type 2 diabetes in youth has accompanied the epidemic of childhood obesity. Moreover, childhood type 2 diabetes is more common in certain ethnic/racial groups. This presentation will discuss the mechanisms underlying the relationship between childhood obesity and risk for type 2 diabetes and identify the metabolic and clinical characteristics of the obese child who may be at higher risk for the disease. The effect of ethnicity on modulating the risk for type 2 diabetes in the obese child is well recognized. Racial differences in the patterns of body fat distribution, glucose homeostasis and substrate utilization are important considerations for the increased risk related to ethnicity and will also be discussed.[br][br]Nothing to Disclose: FFB 2012-06-23T09:30:00 Theater C 2012-06-23T00:00:00 1899-12-30T09:30:00 2523 19 22 S2-1 T07-S03 Saturday 14 2012


2778 ENDO12L_S2-2 SYMPOSIUM SESSION: CLINICAL - Complications of Childhood Obesity (9:30 AM - 11:00 AM) Adolescent BMI as a Predictor of Cornonary Disease [amp] All-Cause Mortality in Midlife Amir Tirosh Brigham [amp] Women[apos]s Hospital and Harvard School of Public Health, Boston, MA It is widely accepted that increased body weight in adolescents is associated with increased cardiovascular disease in midlife, but it is yet unclear whether such an association is independently related to higher body weight early in life or to overweight and obesity in adulthood. Moreover, the applicability of the current definition of overweight and obesity in children and adolescents for prediction of future cardiovascular outcomes is controversial.[br]The Metabolic, Lifestyle and Nutrition Assessment in Young Adult (MELANY) cohort is a prospective study which includes measured height and weight of [gt]40,000 adolescents in Israel with repeated anthropometric, biochemical, social and nutritional assessments into midlife. We followed 37,674 healthy young teenagers with a BMI of 15-36 Kg/m2 at baseline for incident of angiography-proven coronary heart disease (CAD). Height and weight were measured at 3-5 year intervals. During [sim]650,000 person-years (mean follow-up, 17.4 years), we documented 327 new cases of CAD. The study population was divided into deciles based on their age 17 BMI (mean range of 1st to 10th deciles; 17.3-27.6 Kg/m2). In multivariate models adjusted for age, family history, blood pressure, lifestyle factors, and blood biomarkers, elevated adolescent BMI was a significant predictor of angiography-proven CAD (HR, 5.43; 95%CI;2.77-10.62, for the highest vs. the lowest decile). Further adjustment for BMI at adulthood did not attenuate the association with coronary heart disease (HR, 6.85; 95%CI;3.30-14.21), as opposed to the complete reversibility of the risk for type 2 diabetes observed in this population. After adjustment of the BMI values as continuous variables, elevated BMI in both adolescence ([beta]=1.355, P=0.004) and adulthood ([beta]=1.207, P=0.03) were independently associated with CAD (P=0.048 for interaction). BMI values as low as 20.91 Kg/m2 and above were strongly and independently associated with increased incidence of CAD in early adulthood. Additional studies linking data of over 2 million male and female adolescents in Israel since 1950s with all-cause mortality records revealed that BMI at age 17 years at the 75th percentile and above ([sim]23.5 Kg/m2) is independently associated with a gradual increase in mortality rates during 35 years of follow-up.[br]In conclusion, elevated BMI in adolescence, one that is well within the range currently considered to be normal, constitutes a substantial risk factor for CAD and increased mortality in midlife.[br][br](1) Tirosh et. al. NEJM;364(14):1315,2011. (2) Tirosh et. al. Hypertension;56(2):203,2010.[br][br]Sources of Research Support: A grant from the Talpiot Medical Leadership. Program, Chaim Sheba Medical Center, Tel-Hashomer, Israel (to. Dr. Tirosh), and by the Israel Defense Forces Medical Corps. Contact: Atirosh@partners.org.[br][br]Nothing to Disclose: AT 2012-06-23T10:00:00 Theater C 2012-06-23T00:00:00 1899-12-30T10:00:00 2490 19 23 S2-2 T07-S03 Saturday 15 2012


2779 ENDO12L_S2-3 SYMPOSIUM SESSION: CLINICAL - Complications of Childhood Obesity (9:30 AM - 11:00 AM) Non-Alcoholic Fatty Liver Disease [amp] Metabolic Syndrome Valerio Nobili Ospedale Pediatrico Bembino Gesu, Rome, Italy In the last three decades the incidence of metabolic syndrome (MetS) has been growing worldwide along with an increase of obesity, type 2 diabetes mellitus, and non-alcoholic fatty liver disease (NAFLD). In children and adolescents such[br]epidemics are particularly worrisome, since the metabolic consequences in adulthood will signifi cantly burden the health care system.[br]Although the defi nition of MetS in childhood is still controversial, there is agreement with respect to NAFLD being the hepatic manifestation of MetS. However, the molecular pathogenesis of MetS and its contribution to NAFLD is complexand closely related to the pre- and postnatal environment as well as to genetic predisposing factors. The analysis of the possible relationships between NAFLD and MetS is particularly interesting, not only from an epidemiological point of view, but also to better understand the genetic and environmental factors contributing to the development of both diseases.[br][br]Nothing to Disclose: VN 2012-06-23T10:30:00 Theater C 2012-06-23T00:00:00 1899-12-30T10:30:00 2411 19 24 S2-3 T07-S03 Saturday 16 2012


2780 ENDO12L_S3-1 SYMPOSIUM SESSION: BASIC - Intersection of Reward [amp] Metabolic Pathways (9:30 AM - 11:00 AM) Integration of Metabolic Signals by CNS Serotonin Pathways Lora K Heisler University of Cambridge, Cambridge, UK The projected escalation of obesity and the substantial unmet clinical need for pharmacological intervention underscores the necessity of an understanding of the basic neurobiology underlying energy homeostasis. The brain serotonin network has a long established involvement in regulating energy homeostasis, as demonstrated by successful obesity pharmacotherapies targeting this system. However, due to side effects, these compounds have since been withdrawn. An understanding of the precise mechanism through which serotonin modulates energy balance is therefore of intense interest. Pharmacological and genetic research implicates the Gq-coupled 5-HT 2C receptor (5-HT2CR) and the Gi-coupled 5-HT 1B receptor (5-HT1BR) specifically in these effects. We sought to further elucidate the specific mechanism through which d-fenfluramine modulates ingestive behaviour and investigated both downstream and upstream pathways employed. We observed that d-fenfluramine/serotonin activates neurons in the hypothalamic arcuate nucleus (ARC) expressing the endogenous anorectic melanocortin agonist pro-opiomelanocortin (POMC) via a dual mechanism (i) postsynaptic action at 5-HT2CRs expressed on POMC neurons and (ii) presynatpic action at 5-HT1BRs on GABAergic terminals that reduce an inhibitory input onto POMC neurons (1,2). Given this mechanism of action, we then determined that combined 5-HT2CR and 5-HT1BR agonists potentiates appetite suppression compared to single agonist treatment; an effect associated with increased POMC neuron activation on both the population and single cell level. These data further clarify the heterogeneity of ARC POMC neurons by revealing distinct subpopulations activated by 5-HT2CRs, activated by 5-HT1BRs [italic]via [/italic]disinhibiton and regulated by both receptors. Efforts to identify upstream mediators of central serotonin activity revealed a lack of direct modulation by leptin (3). Together, these data identify a critical microcircuit through which 5-HT modulates appetite and reveal multiple points that may be amenable to pharmacological interrogation for future obesity treatment.[br][br]1. Heisler LK, Jobst EE, Sutton GM, Zhou L, Borok E, Thornton-Jones Z, Liu HY, Zigman JM, Balthasar N, Kishi T, Lee CE, Aschkenasi CJ, Zhang CY, Yu J, Boss O, Mountjoy KG, Clifton PG, Lowell BB, Friedman JM, Horvath T, Butler AA, Elmquist JK, Cowley MA. Serotonin reciprocally regulates melanocortin neurons to modulate food intake. Neuron, 2006 51(2):239-49. 2. Xu Y, Jones JE, Kohno D, Williams KW, Lee CE, Choi MJ, Anderson JG, Heisler LK, Zigman JM, Lowell BB & Elmquist JK. 5-HT2CRs expressed by pro-opiomelanocortin neurons regulate energy homeostasis. Neuron, 2008 60(4):582-9. 3. Lam DD, Leinninger GM, Louis GW, Garfield AS, Marston OJ, Leshan R, Scheller EL, Christensen L, Donato J, Xia J, Evans ML, Elias C, Dalley JW, Burdakov D, Myers MG Jr, Heisler LK. Leptin does not directly affect CNS serotonin neurons to influence appetite. Cell Metabolism, 2011 13(5):584-91.[br][br]Sources of Research Support: Work was supported by the Wellcome Trust (WT081713) and NIDDK (DK065171).[br][br]Nothing to Disclose: LKH 2012-06-23T09:30:00 Theater B 2012-06-23T00:00:00 1899-12-30T09:30:00 2473 20 25 S3-1 T04-S05 Saturday 18 2012


2781 ENDO12L_S3-2 SYMPOSIUM SESSION: BASIC - Intersection of Reward [amp] Metabolic Pathways (9:30 AM - 11:00 AM) Lateral Hypothalamic Neurotensin Neurons Integrate Metabolism, Arousal [amp] Motivation Gina Marie Leinninger University of Michigan, Ann Arbor, MI The lateral hypothalamic area (LHA) contains a variety of neurons that integrate metabolic signals with the appropriate behavioral outputs, many of which are ultimately regulated by the mesolimbic dopamine system. The physiological roles of some LHA neuronal populations are beginning to be understood, such as neurons containing the neuropeptide melanin-concentrating hormone (MCH) or a separate population expressing orexin/hypocretin (OX/Hcrt) that are important regulators of alertness, locomotor activity and reward. We have begun to characterize another large population of LHA neurons: GABAergic neurons that co-express the neuropeptide Neurotensin (Nts); we refer to these as LHA Nts neurons. LHA Nts neurons are activated by diverse metabolic stimuli, including leptin, dehydration or inflammatory signals, but these signals inhibit adjacent OX/Hcrt neurons. Indeed, LHA Nts neurons project to the ventral tegmental area (VTA) as well to OX/Hcrt neurons, which themselves project to the VTA. LHA Nts neurons therefore represent a novel pathway to integrate metabolic sensing with control of OX/Hcrt-mediated behaviors and the mesolimbic dopamine system, including regulation of alertness and locomotor activity. Utilizing mouse models that enable selective interrogation of LHA Nts neurons, including the subpopulation of LHA Nts neurons that are leptin-responsive, we are beginning to understand the neural and behavioral mechanisms by which Nts neurons regulate OX/Hcrt neurons and contribute to energy balance.[br][br]Nothing to Disclose: GML 2012-06-23T10:00:00 Theater B 2012-06-23T00:00:00 1899-12-30T10:00:00 2466 20 26 S3-2 T04-S05 Saturday 19 2012


2782 ENDO12L_S3-3 SYMPOSIUM SESSION: BASIC - Intersection of Reward [amp] Metabolic Pathways (9:30 AM - 11:00 AM) Integration of Peripheral Signals by MCH Neurons Eleftheria Maratos-Flier Beth Israel Deaconess Medical Center, Boston, MA Disclosure Incomplete: EM-F 2012-06-23T10:30:00 Theater B 2012-06-23T00:00:00 1899-12-30T10:30:00 6049 20 27 S3-3 T04-S05 Saturday 20 2012


2783 ENDO12L_S4-1 SYMPOSIUM SESSION: TRANSLATIONAL - New Developments in Steroid Metabolism in Cancer (9:30 AM - 11:00 AM) Breast Cancer: Obesity [amp] Estrogen Metabolism Kristy A Brown Prince Henry[apos]s Institute, Clayton, Australia The risk of postmenopausal breast cancer is increased two-fold in obese women, and the majority of postmenopausal and obesity-related breast cancers are estrogen-dependent. The adipose tissue is the major source of estrogens in postmenopausal women and local production of estrogens within breast adipose stromal cells (ASCs) is known to drive tumour growth. Our laboratory[apos]s work has focussed on understanding how dysregulated metabolism, as is seen in obesity and cancer, increases estrogen production within the breast. In this regard, we have studied a number of metabolic pathways, including those which modulate the activity of CRTC2 and hypoxia inducible factor-1[alpha] (HIF1[alpha]), for their role in regulating a key regulator of estrogen biosynthesis, aromatase.[br]AMPK is a master regulator of energy homeostasis. Changes in nutrient availability cause its activation by upstream kinases, including LKB1, which leads to the stimulation of pathways of energy production and inhibition of pathways of energy utilisation. Our previous work has demonstrated that LKB1/AMPK are negative regulators of aromatase expression in the ASCs via cytoplasmic sequestration of the CREB-coactivator CRTC2. We have also shown that tumour-derived inflammatory factors, including prostaglandin E2 (PGE2), and obesity-derived factors, including leptin, inhibit the expression and activity of LKB1 and AMPK. As a result, CRTC2 enters the nucleus, binds to the proximal promoter PII of aromatase via cAMP response elements (CRE) and increases its expression. Conversely, adiponectin, produced by lean adipose tissue, stimulates LKB1/AMPK and inhibits the PGE2-mediated expression of aromatase.[br]We have recently identified a putative hypoxia response element which overlaps the proximal CRE of aromatase PII. HIF1[alpha] has an established role in the vascularisation of tumours and is emerging as a key mediator of metabolic responses in cancer. We now demonstrate that HIF1[alpha] is a stimulator of aromatase expression within ASCs and we have found that PGE2 stimulates the expression of HIF1[alpha] and leads to its stabilisation in the nucleus of ASCs. We also show that HIF1[alpha] binds directly to aromatase promoter PII and stimulates its expression.[br]Taken together, our work suggests that dysregulated metabolism is not only a characteristic of adipocytes or the tumorous epithelium, but also of the breast adipose stroma, and that aromatase expression is tightly regulated by these metabolic pathways in obesity and breast cancer.[br][br]Nothing to Disclose: KAB 2012-06-23T09:30:00 Room 351 2012-06-23T00:00:00 1899-12-30T09:30:00 2457 21 28 S4-1 T08-S09 Saturday 22 2012


2784 ENDO12L_S4-2 SYMPOSIUM SESSION: TRANSLATIONAL - New Developments in Steroid Metabolism in Cancer (9:30 AM - 11:00 AM) Lung Cancer: An Estrogen Target? Yasuhiro Miki Tohoku University, Sendai, Japan Estrogen plays an important role in the progression of breast cancer and a majority of the human breast cancers express the estrogen receptor. estrogen receptors especially estrogen receptor [beta] have been demonstrated to be present and functional in both normal human lung and its disorders including cancer. Non-small cell lung carcinomas (NSCLC) are well-known to be composed of heterogeneous groups. Squamous cell carcinoma is the most common subtype in men, but adenocarcinoma is the most common histologic subtype in women. Therefore, sex steroid hormones such as estrogens have been considered to play some roles in NSCLC. Results of several epidemiological analyses pointed out the association between physiological or artificial alterations of hormone status such as menstruation and postmenopausal administration of hormone replacement therapy and lung cancer risks or its development especially in female subjects. In NSCLC tissues, intratumoral estrogen synthesis via aromatase, which is a key enzyme in the estrogen synthesis involved in aromatization of androgens (androsetedione and testosterone) into estrogens (estrone and estradiol), has recently become of clinical interest as a possible target of therapy. Several studies demonstrated that immunoreactivity of aromatase was detected in carcinoma cells of approximately over 60[ndash]70% lung carcinoma cases. Aromatase has been reported to be predominantly expressed in intratumoral stromal cells of human breast carcinoma tissues. In lung carcinoma, however, several studies showed aromatase protein in parenchymal cells but not in stromal cells. We reported that the intratumoral estradiol level was significantly associated with the tumor size and cell proliferation activity in 59 NSCLC tissues. Furthermore, male NSCLC cases with double positivity for aromatase and estrogen receptor [beta] demonstrated lower status in N factor by TNM classification. These findings encourage the use of ER and aromatase inhibitors in NSCLC patients.[br][br]Nothing to Disclose: YM 2012-06-23T10:00:00 Room 351 2012-06-23T00:00:00 1899-12-30T10:00:00 2396 21 29 S4-2 T08-S09 Saturday 23 2012


2785 ENDO12L_S4-3 SYMPOSIUM SESSION: TRANSLATIONAL - New Developments in Steroid Metabolism in Cancer (9:30 AM - 11:00 AM) Prostate Cancer: Cutting Off Androgens at the Pass Nima Sharifi University of Texas Southwestern, Dallas, TX Gonadal testosterone (T) is converted by steroid-5[alpha]-reductase to dihydrotestosterone (DHT), which drives tumor progression through the androgen receptor (AR) in prostate cancer. Following initial treatment responses to gonadal T deprivation, advanced tumors progress as castration-resistant prostate cancer (CRPC). The underlying mechanisms that drive CRPC involve the genesis of DHT to concentrations that are abundantly sufficient to bind and activate the androgen receptor (AR). There are several possible routes to the synthesis of DHT and the major pathway from adrenal precursors that allows resistance to gonadal T deprivation circumvents T and instead requires 5[alpha]-reduction of [Delta][sup]4[/sup]-androstenedione to 5[alpha]-androstanedione. The role and requirement of DHT in driving CRPC is made even clearer by the survival benefit conferred by CYP17A1 inhibition with abiraterone acetate and potent AR antagonism with MDV3100. Although there are variations in clinical response to these newer agents, resistance again eventually occurs. Resistance to these therapies may yet remain dependent on steroidogenesis. However, just as the 5[alpha]-androstanedione pathway is required for DHT synthesis in CRPC, other noncanonical pathways may be involved in resistance to abiraterone acetate and MDV3100. The elucidation of the pathways involved is a necessary step to the identification of the next generation of pharmacologic targets and therapies.[br][br]Sources of Research Support: Howard Hughes Medical Institute, Prostate Cancer Foundation, American Cancer Society, CDMRP Prostate Cancer Research Program.[br][br]Nothing to Disclose: NS 2012-06-23T10:30:00 Room 351 2012-06-23T00:00:00 1899-12-30T10:30:00 2445 21 30 S4-3 T08-S09 Saturday 24 2012


2786 ENDO12L_S5-1 SYMPOSIUM SESSION: BASIC - New Insights into Signaling by GPCR[apos]s (9:30 AM - 11:00 AM) Agonist-Dependent Control of GPCR Function by Site-Specific Phosphorylation Agnes Schonbrunn University of Texas Southwestern Medical Center, Houston, TX Hormone binding to G protein coupled receptors leads both to G-protein activation and to receptor phosphorylation on multiple intracellular Ser and Thr residues by second messenger activated and G protein coupled receptor kinases. Once phosphorylated, activated receptors recruit arrestins, cytoplasmic proteins that sterically inhibit receptor-G protein coupling thereby desensitizing G-protein mediated signalling and protecting cells from overstimulation. In addition, because arrestins bind to many proteins involved in receptor internalization or intracellular signaling, the recruitment of arrestins by phosphorylated GPCRs leads both to the endocytosis of cell surface receptors and to the initiation of G protein-independent signaling events. Thus, multi-site phosphorylation determines the signaling capacity as well as the subcellular distribution of GPCRs as part of an exquisitely regulated process. Recent studies have suggested that multi-site GPCR phosphorylation coordinates receptor signaling and trafficking by providing a bar-code that affects which proteins are recruited to the receptor and how these proteins function after receptor binding, analogous to other phosphorylation-dependent scaffolding interactions involved in signal transduction. Moreover, this bar-code is determined by the nature of the receptor-bound ligand as well as by the tissue environment of the receptor. Thus, biased agonists that preferentially direct a receptor towards one signaling modality over another can produce a unique pattern of receptor phosphorylation and thus confer distinct properties to receptors that can be exploited for drug development. In the case of one specific GPCR, somatostatin receptor 2A (sst2A), somatostatin binding stimulates receptor phosphorylation on multiple identified residues in the carboxy-terminus (1,2). Site directed mutagenesis of sst2A phosphorylation sites showed that different phosphorylated residues mediate receptor desensitization from those required for receptor internalization (3)- a characteristic feature of phosphorylation bar-codes. Strikingly, pasireotide (SOM230), a biased agonist (4), produces a different pattern of sst2A receptor phosphorylation from that elicited by the native hormone, consistent with pasireotide[apos]s compromised ability to induce sst2A receptor internalization (5). I will discuss how the sst2A receptor phosphorylation bar-code is produced, how it is regulated, and how it affects receptor function.[br][br]Madhumita Ghosh, Yachu J. Kao, Qisheng Liu and Kimberly A. Loesch in our group contributed to the projects discussed. (1) Liu Q et al Molecular Pharmacology 2009; 76:68. (2) Ghosh M and Schonbrunn A 2011; J Biol Chem 286:13561. (3) Liu Q et al Molecular Pharmacology 2008; 73:292. (4) Cescato R et al Mol Endocrinol 2010; 24:240-9. (5) Kao YJ et al 2011 Mol Endocrinol 25:1040.[br][br]Sources of Research Support: NIH Grant DK032234 awarded to AS; Pharmacoinformatics. Training Program of the Keck Center of the Gulf. Coast Consortia (NIH Grant T90 DK070109) awarded to KAL.[br][br]Nothing to Disclose: AS 2012-06-23T09:30:00 Room 361 ABDE 2012-06-23T00:00:00 1899-12-30T09:30:00 2538 22 31 S5-1 T08-S07 Saturday 26 2012


2787 ENDO12L_S5-2 SYMPOSIUM SESSION: BASIC - New Insights into Signaling by GPCR[apos]s (9:30 AM - 11:00 AM) Roles of Endocannabinoids in G Protein-Coupled Receptor Signaling Laszlo Hunyady Semmelweis University, Budapest, Hungary It has been shown that diacylglycerol (DAG) formed during the activation of Gq-coupled receptors can be converted to 2-arachidonoylglycerol (2-AG), an important endocannabinoid, by DAG lipase in the central nervous system and in peripheral tissues. We have studied the effect of AT1 angiotensin receptors (AT1Rs) on CB1 cannabinoid receptor (CB1R) activity in CHO cells by detecting bioluminescence resonance energy transfer (BRET) between subunits of Go proteins fused to Renilla luciferase or yellow fluorescent protein. AT1R stimulation with angiotensin II (Ang II) caused activation of CB1Rs. This effect was also observed, when the two receptors were expressed in separate set of cells and mixed prior to the experiment, suggesting that endocannabinoid release causes paracrine activation of CB1R. These effects of AT1R activation were mediated by DAG lipase-mediated accumulation of 2-AG. Paracrine transactivation of CB1R also occurred during the stimulation of M1, M3, M5 muscarinic, [alpha]1-adrenergic, B2 bradykinin or V1 vasopressin receptors co-expressed with CB1R. To study the role of this mechanism in the regulation of vascular tone, isolated rat and mouse gracilis arterioles and mouse saphenous arteries were studied by microangiometry. In these vessels CB1R-inhibition augmented the Ang II-induced vasoconstriction by 30-80 %, suggesting that Ang II-induced endocannabinoid release modulates its vasoconstrictor action. In another set of experiments, the central hypertensive effect of Ang II, injected directly into the hypothalamic paraventricular nucleus of anaesthetized rats, was abolished by AM251, an inverse agonist of CB1Rs. These results suggest that stimulation of AT1Rs or other Gq-coupled receptors leads to 2-AG release and paracrine transactivation of CB1Rs, which can modulate the peripheral and central hypertensive effects of Ang II and other calcium-mobilizing hormones.[br][br]Co-authors:. Maria Szekeres1, Gyorgy L. Nadasy2, Eszter Soltesz-Katona1, Gyorgy Bagdy3, Gabor Turu1. Departments of 1Physiology and 2Human Physiology and Clinical Experimental Research, Faculty of Medicine; and 3Department of Pharmacodynamics, Faculty of Pharmacy, Semmelweis University, Budapest, Hungary.[br][br]Sources of Research Support: Hungarian National Grant OTKA NK-100883; T[Aacute]MOP (4.2.1.B-09/1/KMR-2010-0001).[br][br]Nothing to Disclose: LH 2012-06-23T10:00:00 Room 361 ABDE 2012-06-23T00:00:00 1899-12-30T10:00:00 2472 22 32 S5-2 T08-S07 Saturday 27 2012


2788 ENDO12L_S5-3 SYMPOSIUM SESSION: BASIC - New Insights into Signaling by GPCR[apos]s (9:30 AM - 11:00 AM) GPCR Complexes [amp] Their Novel Pharmacology Kevin Donald George Pfleger WA Institute for Medical Research, Nedlands Perth WA, Australia It has become clear that GPCRs function in macromolecular complexes that may constitutively or dynamically include G proteins, kinases, arrestins, adaptor proteins, ubiquitin, other GPCRs and potentially a plethora of other proteins. The pharmacology of a GPCR can therefore vary considerably depending upon which other proteins are present in the same cell and its propensity to functionally interact with them. Consequently, our understanding of GPCR function, is being enhanced by the development and application of technological approaches that enable monitoring of receptor complexes in live cells and in real-time, such as those based on bioluminescence resonance energy transfer (BRET). As one example, we have observed differences between orexin receptor 1 and 2 complexes and their propensity to interact with arrestins and ubiquitin using extended BRET temporal profiling, as well as observing correlations with signaling via ERK1/2. Our recent work indicates that differences in specific C-terminal clusters of serines and threonines dictate the differences between these receptors, consistent with the concept of phosphorylation [apos]bar codes[apos]. We have also observed novel pharmacology as a consequence of GPCR heteromerization, such as between the [alpha]1A-adrenoceptor ([alpha]1A-AR) and the CXC chemokine receptor 2 (CXCR2) that we believe may have implications in the prostate. Norepinephrine does not normally induce robust recruitment of arrestins to the [alpha]1A-adrenoceptor in human embryonic kidney 293 cells, but does when CXCR2 is co-expressed. Such heteromer-specific novel pharmacology also has the potential to unmask biased signaling, as we have observed with Labetalol acting at the [alpha]1A-AR/CXCR2 heteromer.[br][br]Sources of Research Support: Australian Research Council; National Health and Medical Research Council of Australia; Dimerix Bioscience Pty Ltd.[br][br]Disclosures: KDGP: Chief Scientific Officer, Dimerix Bioscience Pty Ltd. 2012-06-23T10:30:00 Room 361 ABDE 2012-06-23T00:00:00 1899-12-30T10:30:00 2480 22 33 S5-3 T08-S07 Saturday 28 2012


2789 ENDO12L_S6-1 SYMPOSIUM SESSION: BASIC - Pituitary Cells as Metabolic Sensors (9:30 AM - 11:00 AM) Somatotropes as Metabolic Sensors Rhonda Denise Kineman Jesse Brown VA Medical Center/University of Chicago-Illinois, Chicago, IL Weight gain lowers circulating GH levels. Since GH is lipolytic and anabolic, the decline in GH observed with weight gain may exacerbate metabolic dysfunction. The mechanism(s) causing GH levels to fall with weight gain remains a subject of debate. However, evidence suggests hyperinsulinemia plays a key role. This hypothesis is based on the fact that GH release is negatively associated with insulin levels, where weight loss sufficient to lower insulin levels restores GH output. Insulin may work centrally to modify hypothalamic input to the pituitary somatotrope and/or act directly at the level of the pituitary to suppress somatotrope function. We are interested in determining if the pituitary somatotrope is a (patho)physiologic target of insulin. In order to investigate insulin[apos]s role, one must also consider the actions of IGF-I since: 1) IGF-I is a primary negative feedback signal controlling GH secretion, 2) INSR and IGFIR are structurally/functionally related, therefore high insulin levels can activate IGFIR, 3) the pituitary receptors for insulin (INSR) and IGF-I (IGFIR) are expressed at comparable levels; and 4) INSR and IGFIR may interact by forming hybrid receptors or via crosstalk of downstream intracellular signaling pathways, thereby modifying the response to their respective ligands. In order to tease apart these complicated systems we have used the Cre/loxP system to inactivate the somatotrope INSR or/and IGFIR, [italic]in vitro[/italic] and [italic]in vivo[/italic]. Several novel concepts have emerged from this series of studies. First, although insulin and IGF-I ultimately inhibit GH secretion, their [italic]in vitro[/italic] and [italic]in vivo[/italic] mechanisms of action are distinct. Second, somatotrope-specific loss of INSR or/and IGFIR increases GH levels, which cannot be compensated for by an intact hypothalamic feedback system, thereby establishing the somatotrope as a primary sensor for changes in circulating insulin/IGF-I. Third, under lean conditions, somatotrope-specific loss of INSR has a more profound effect on circulating GH/IGF-I than loss of IGFIR, suggesting insulin plays a primary role in regulating short-term changes in GH secretion. Finally, in the context of diet-induced obesity, the direct effects of insulin (or IGF-I) on somatotrope function cannot fully account for the fall in GH levels and therefore must be driven by insulin-induced changes in hypothalamic function or other systemic factors that act centrally or directly on the somatotrope to reduce GH output.[br][br]Sources of Research Support: Veteran Affairs Merit Award BX001114; NIH DK088133 awarded to RDK.[br][br]Nothing to Disclose: RDK 2012-06-23T09:30:00 Room 371 2012-06-23T00:00:00 1899-12-30T09:30:00 2528 23 34 S6-1 T01-S07 Saturday 30 2012


2790 ENDO12L_S6-2 SYMPOSIUM SESSION: BASIC - Pituitary Cells as Metabolic Sensors (9:30 AM - 11:00 AM) AMPK: Integrating Energy Balance [amp] Fertility in Gonadotropes Joelle Dupont Institute National de la Recherche Agro, Nouzilly, France AMP-activated protein kinase (AMPK) is a serine/threonine protein kinase, which serves as a fuel sensor in regulating energy metabolism in many cell types. It is activated by phosphorylation when a cellular stress increases the AMP:ATP ratio due to limited generation of ATP or increased ATP depletion and, consequently, AMP production. In addition, it is also activated by several hormones and cytokines such as adiponectin, leptin and ghrelin, and by the antidiabetic drug metformin. Little is known about the regulation and the effects of AMPK in the anterior pituitary. To our knowledge, the sole papers reporting on the modulation of AMPK activity in pituitary cells in response to hormones or metabolic stressors suggest a role of AMPK in regulating luteinizing hormone (LH) secretion in gonadotrope cells, ACTH precursor expression in corticotroph cells and somatotroph cell function. In our laboratory, we have shown that the different subunits of AMPK are expressed in rat primary pituitary cells and in the mouse gonadotrope cell line, LBetaT2 cells. Furthermore we have observed that activation of AMPK by metformin can directly impact the signaling pathways involved in gonadotropin synthesis and secretion. More precisely, we have shown that in rat pituitary cells, AMPK activation induced by metformin treatment decreases gonadotropin release induced by GnRH and activin and this mechanism is associated with inhibition of the MAPK ERK1/2 and SMAD2 signaling pathways, respectively.[br]Thus, AMPK could be one of the signaling pathways controlling the interactions between energy balance and reproductive functions at the pituitary levels. The reproductive system is tightly coupled with energy balance, and thereby metabolic abnormalities can lead to the development of some alterations in fertility such as the polycystic ovary syndrome (PCOS). Some studies indicate that metformin treatment in PCOS patient is also able to decrease LH secretion. Since AMPK is involved in the mechanism of action of metformin at the pituitary levels, it appears as an ideal target for the development of novel therapies.[br][br]Nothing to Disclose: JD 2012-06-23T10:00:00 Room 371 2012-06-23T00:00:00 1899-12-30T10:00:00 2400 23 35 S6-2 T01-S07 Saturday 31 2012


2791 ENDO12L_S6-3 SYMPOSIUM SESSION: BASIC - Pituitary Cells as Metabolic Sensors (9:30 AM - 11:00 AM) Insulin Suppression of LH in PCOS Mark A Lawson University of California-San Diego, La Jolla, CA Polycystic Ovarian Syndrome is generally characterized by elevated androgen, luteinizing hormone (LH), and insulin levels, as well as other factors. Inappropriate gonadotropin secretion has been attributed to altered gonadotropin-releasing hormone (GnRH) pulsatility, disrupted regulatory feedback, or to other endocrine factors contributing to the reproductive endocrine axis. Association of elevated insulin levels and the predisposition of women with PCOS toward metabolic syndrome or type II DM led to the hypothesis that insulin input into the reproductive endocrine axis could promote inappropriate gonadotropin secretion. This was supported by the ability of insulin to promote LH secretion in primary pituitary cell culture. However, the complex endocrine profiles of women with PCOS has made it difficult to correlate insulin, androgen, or other factors with LH or to use these to develop a useful descriptive model from which predictions of endocrine action could be made or potential interventions be developed.[br]We developed a descriptive model of mean 12h LH by considering multiple endocrine and physiological parameters in both normal and PCOS women. Using this approach, we identified three independent factors correlating with mean LH; testosterone, insulin, and body mass index (BMI). Androgen was positively correlated with elevated LH in this model whereas insulin and BMI were negatively correlated. Using the euglycemic-hyperinsulinemic clamp, we directly examined the effect of elevated insulin on baseline LH and pituitary sensitivity by measuring excursion from baseline and area under the curve after GnRH challenge. These studies confirmed the predicted suppressive effect of insulin on LH secretion. Overall the correlation of these factors and the effect of insulin were most pronounced in women with PCOS.[br]Further examination of insulin action in vitro using a murine gonadotrope cell model has yielded more complex results. We and others have shown that an insulin signaling system exists in gonadotropes but differences from other tissues are apparent. Insulin effects on LH protein synthesis and [italic]Lhb[/italic] gene expression are dependent on the context of insulin and GnRH exposure. Overall, we conclude that insulin has direct input to the reproductive axis at the pituitary and may inform the axis on metabolic or energy status. We are pursuing factors related to increased BMI to discern their effect on gonadotrope function in both normal and PCOS populations.[br][br]Sources of Research Support: NIH Grants R21 DK68167, R01 HD43758, MO1 RR00827, and U54HD012303.[br][br]Nothing to Disclose: MAL 2012-06-23T10:30:00 Room 371 2012-06-23T00:00:00 1899-12-30T10:30:00 2515 23 36 S6-3 T01-S07 Saturday 32 2012


2792 ENDO12L_S7-1 SYMPOSIUM SESSION: TRANSLATIONAL - Prolactin [amp] Cancer (9:30 AM - 11:00 AM) Targeting the Prolactin Receptor in Ovarian Cancer Ameae M Walker University of California, Riverside, CA Standard treatment of ovarian cancer involves surgical de-bulking and chemotherapy, but even with this treatment 20-30% of patients never experience a period of remission. Even among those who do, the majority will eventually relapse, making it the most lethal of the gynecological cancers. Clearly, better treatment regimens are needed. Recent reports have suggested a relationship between prolactin (PRL) levels and ovarian cancer (1). Furthermore, in a small sampling, 90% of primary human ovarian cancers were shown to express PRL receptors (PRLR) (2). Our functional analysis of the role of PRL has used three ovarian cancer lines as examples derived from an ascites (OV90), a clear cell adenoma (TOV21G) and an endometrioid adenoma (TOV112D). Each cell line expresses both PRL and PRLRs, and uses secreted PRL as an autocrine growth/survival factor. We therefore tested the potential therapeutic effects of two PRLR antagonists. PRL promoted, and each antagonist reduced, cell growth, survival, and migration in each cell line. The disease-promoting effects of PRL were enhanced by increased expression of the long form (LF) PRLR and antagonized by increased expression of SF1b PRLR. However, increased expression of SF1b was not the mechanism by which the receptor antagonists achieved their beneficial effects. BRCA1 mutations increase risk of ovarian cancer, illustrating the importance of this tumor suppressor. We therefore examined the effect of PRL and one of the antagonists on the BRCA1-p21 axis. Surprisingly, both PRL and the antagonist promoted BRCA1 expression, nuclear entry, and binding to the p21 promoter. However with PRL, binding to the p21 promoter did not result in activation, whereas with the antagonist, there was increased p21 expression. Further analysis demonstrated that phosphorylation of Stat5 by PRL, which occurs only at the LF PRLR, produced a complex between phospho-Stat5 and BRCA1 that was inactive on the p21 promoter. The antagonist blocked phosphorylation of Stat5 in response to autocrine as well as added PRL. Knockdown of the LF PRLR decreased cell number. PRL also increased phosphorylation of p21, the result of which is retention of p21 in the cytosol where it is anti-apoptotic. By contrast, the antagonist decreased phosphorylation of p21, allowing nuclear entry and inhibition of the cell cycle/promotion of apoptosis. We conclude that PRLR antagonists and agents that reduce LF PRLR may have utility as therapeutics in this devastating disease.[br][br](1)Mor G et al., Proc Nat Acad Sci USA 2005; 102: 7677. (2)Asai-Sato M et al., Int J Cancer 2005; 115: 539.[br][br]Sources of Research Support: DOD 0C073294.[br][br]Nothing to Disclose: AMW 2012-06-23T09:30:00 Room 342 ABDE 2012-06-23T00:00:00 1899-12-30T09:30:00 2444 24 37 S7-1 T06-S03 Saturday 34 2012


2793 ENDO12L_S7-2 SYMPOSIUM SESSION: TRANSLATIONAL - Prolactin [amp] Cancer (9:30 AM - 11:00 AM) Targeting Prolactin Signaling in Prostate Cancer Marja T Nevalainen Thomas Jefferson University, Philadelphia, PA Organ-confined prostate cancer (PCa) is treated by surgery or radiation, while options for disseminated PCa include hormone therapy, radiation, or chemotherapy. Response to hormone therapy is of limited duration and metastatic PCa inevitably becomes castrate-resistant (CR), a stage for which there is no cure. The molecular mechanisms underlying progression of PCa to castrate-resistant and metastatic disease are poorly understood. Prolactin (Prl) is a mitogen and survival factor for androgen-deprived prostate cancer. Signal Transducer and Activator of Transcription 5a/b (Stat5a/b) are the key molecules mediating the biological effects of Prl in prostate cancer. Stat5a/b is critical for PCa cell viability and PCa xenograft tumor growth. While Stat5a/b physically interacts with androgen receptor (AR) in PCa cells and promotes nuclear localization and transcriptional activity of the AR, inhibition of Stat5a/b induces extensive apoptotic death of both AR-positive and AR-negative PCa cells suggesting that Stat5a/b acts through AR-dependent and AR-independent molecular mechanisms. Stat5a/b induces metastatic behavior of human PCa cells [italic]in vitro[/italic] and in experimental metastases assays [italic]in vivo[/italic]. Nuclear Stat5a/b protein levels are increased in high grade PCas, in CR PCas and distant metastases. High nuclear Stat5a/b expression predicts early disease recurrence and PCa-specific death in intermediate grade primary human PCas. Based on these findings, Stat5a/b represents a therapeutic target protein for advanced PCa. Progress with small-molecule inhibitors of Jak2 or Stat5a/b in blocking castrate-resistant PCa xenograft tumor growth and in induction of apoptotic death of clinical prostate cancers [italic]ex vivo[/italic] in explant organ cultures will be presented.[br][br]Sources of Research Support: NCI 1RO1CA113580-01A1. American Cancer Society RSG-04-196-01-MGO. Department of Defense W81XWH-05-01-0062, W81XWH-06-01-0076, W81XWH-07-1-0411.[br][br]Nothing to Disclose: MTN 2012-06-23T10:00:00 Room 342 ABDE 2012-06-23T00:00:00 1899-12-30T10:00:00 2535 24 38 S7-2 T06-S03 Saturday 35 2012


2794 ENDO12L_S7-3 SYMPOSIUM SESSION: TRANSLATIONAL - Prolactin [amp] Cancer (9:30 AM - 11:00 AM) Prolactin [amp] Breast Cancer: Insights from Mouse Models Linda A Schuler University of Wisconsin, Madison, WI Prolactin (PRL) is critical for mammary alveolar morphogenesis and differentiation. Accumulating epidemiological studies have also implicated PRL in the development of particularly ER[alpha]+ breast cancer, and cancer progression, metastases, therapeutic resistance and poor survival. Multiple ligands, including circulating PRL and growth hormone, from multiple sources, including both pituitary-produced and locally synthesized PRL, can activate PRL receptors in the human breast. Our murine transgenic model, NRL-PRL, mimics this local mammary PRL production, offering the opportunity to examine the actions of this hormone on breast pathology in the dynamic[italic] in vivo[/italic] environment. Increased mammary exposure to PRL results in diverse aggressive adenocarcinomas, independent of ovarian hormones. Many of these carcinomas express ER[alpha] and features of the luminal subtype of human breast cancer. Although early lesions are responsive to estrogen, advanced carcinomas are insensitive despite continued ER[alpha] expression, modeling anti-estrogen resistant tumors. In combination with other genetically modified models, we have examined interactions of PRL with growth factors, oncogenic mutations and intracellular mediators in breast pathogenesis. Together, our findings provide insight into the pathogenesis of luminal tumors, responsiveness to estrogens and PRL actions in this disease. They support therapeutic targeting of PRL signals alone or in combination with other therapies for breast cancer.[br][br]Nothing to Disclose: LAS 2012-06-23T10:30:00 Room 342 ABDE 2012-06-23T00:00:00 1899-12-30T10:30:00 2509 24 39 S7-3 T06-S03 Saturday 36 2012


2795 ENDO12L_S8-1 SYMPOSIUM SESSION: BASIC - Thyroid Hormone [amp] Energy Expenditure (9:30 AM - 11:00 AM) Local Control of Thyroid Hormone Action Via the Deiodinases Stephen Huang Children[apos]s Hospital Boston, Boston, MA Disclosure Incomplete: SH 2012-06-23T09:30:00 Room 320 2012-06-23T00:00:00 1899-12-30T09:30:00 6027 25 40 S8-1 T02-S10 Saturday 38 2012


2796 ENDO12L_S8-2 SYMPOSIUM SESSION: BASIC - Thyroid Hormone [amp] Energy Expenditure (9:30 AM - 11:00 AM) Thyroid Hormone Action on the Nervous System Bjorn Vennstrom Karolinska Institute, Solna, Sweden Disclosure Incomplete: BV 2012-06-23T10:00:00 Room 320 2012-06-23T00:00:00 1899-12-30T10:00:00 6028 25 41 S8-2 T02-S10 Saturday 39 2012


2797 ENDO12L_S8-3 SYMPOSIUM SESSION: BASIC - Thyroid Hormone [amp] Energy Expenditure (9:30 AM - 11:00 AM) Selective Modulation of Thyroid Hormone Signaling [amp] Brown Fat Paul Webb Methodist Hospital Research Institute, Houston, TX Obesity is arguably the worst health problem affecting the western world. Current approaches to treatment of obesity and associated diseases such as type 2 diabetes mellitus (T2DM) have not reduced these epidemics; in spite of aggressive efforts to modify lifestyles in the United States; success will require many years. It is essential to develop innovative treatments for these disorders. Thyroid hormones (THs) and their cognate nuclear receptors (TRs [alpha] and [beta]) are master regulators of metabolism and TH excess leads to deleterious effects such as elevated heart rate, arrythmias and muscle and bone catabolism along with beneficial effects such as reduced cholesterol and increased fat burning leading to fat loss. Our team developed selective TR modulators (STRMs) to safely elicit beneficial effects of TH excess without harmful effects; STRMs are TR[beta] selective, engineered into ligands because TR[beta] mediates cholesterol lowering, and exhibit adventitious liver selective uptake and limited gene selectivity. STRMs show promise for treatment of elevated serum cholesterol in animals and at least three have entered human clinical trials for dyslipidemias. However, some STRMs exhibit other potentially beneficial effects in rodents, including reduction of adipose tissue mass, reductions in blood glucose in diabetic animals and reversal of non-alcoholic fatty liver disease (NAFLD). We show that rapid weight loss is achieved safely in short periods of STRM treatment and that it is closely linked to activation of brown adipose tissue (BAT) thermogenic activity. Reduction of blood glucose is also linked to BAT activation. Comparison of modestly and highly liver-selective ligands (GC-1 and KB2115) suggests that fat loss and glucose-reductions require extra-hepatic actions and we show that GC-1 and similar compounds rescue a defect in thermogenic response in obese rodents via direct effects on BAT. In contrast, both GC-1 and KB2115 reverse NAFLD suggesting that this involves direct effects on liver. This action is not connected with insulin sensitization; in fact, highly liver selective STRMs may stimulate gluconeogenesis and worsen insulin response. We conclude that different STRMs are best suited for different aspects of metabolic disorders, that STRMs could be novel therapeutics to treat obesity and NAFLD and that some STRMs could be used to reduce elevated blood glucose in a manner that is completely independent of insulin.[br][br]Sources of Research Support: This work was supported by NIH RC4 DK090849-01. Optimal Selective Thyroid Hormone Analogs for Metabolic Syndrome.[br][br]Nothing to Disclose: PW 2012-06-23T10:30:00 Room 320 2012-06-23T00:00:00 1899-12-30T10:30:00 2534 25 42 S8-3 T02-S10 Saturday 40 2012


2798 ENDO12L_S9-1 SYMPOSIUM SESSION: TRANSLATIONAL - Treatment Challenges in Osteoporosis (9:30 AM - 11:00 AM) Matching Therapy To Fit the Patient Felicia Cosman Helen Hayes Hospital, West Haverstraw, NY Disclosure Incomplete: FC 2012-06-23T09:30:00 Theater A 2012-06-23T00:00:00 1899-12-30T09:30:00 6036 26 43 S9-1 T03-S04 Saturday 42 2012


2799 ENDO12L_S9-2 SYMPOSIUM SESSION: TRANSLATIONAL - Treatment Challenges in Osteoporosis (9:30 AM - 11:00 AM) When to Stop & Start Anti-Resorptive Therapy Richard S Bockman Hosp for Special Surgery, New York, NY 2012-06-23T10:00:00 Theater A 2012-06-23T00:00:00 1899-12-30T10:00:00 6286 26 44 S9-2 T03-S04 Saturday 43 2012


2800 ENDO12L_S9-3 SYMPOSIUM SESSION: TRANSLATIONAL - Treatment Challenges in Osteoporosis (9:30 AM - 11:00 AM) Treatment Challenges in Osteoporosis: Mechanisms of Action of Emerging Therapies Roland E Baron Harvard School of Dental Medicine, Boston, MA The therapeutic options for the treatment of osteoporosis have so far comprised mostly anti-resorptive drugs, in particular bisphosphonates and more recently RANKL antibodies (denosumab). These drugs have, however, limitations and in particular the fact that they lead to a low turnover state where bone formation decreases with the decrease in bone remodeling activity. Novel anti-resorptives (odanacatib) may have a different profile with effective inhibition of bone resorption associated with minimal reduction in bone formation and bone turnover and, possibly even some cortical stimulation of bone formation. An alternative class of osteoporosis drugs, i.e. bone anabolics have recently reached the market or clinical development. We will discuss their biology and the perspectives they offer for our therapeutic armamentarium. We will focus on the two main osteo-anabolic pathways identified as of today: parathyroid hormone (PTH), the only anabolic drug currently on the market, and activation of canonical Wnt signaling through inhibition of the endogenous inhibitors sclerostin and dickkopf1 (Dkk1). Each approach is based on a different molecular mechanism but most recent evidence suggests that these two pathways may actually converge, at least in part. Whereas recombinant human PTH (rhPTH) treatment is being revisited with different formulations and attempts to regulate endogenous PTH secretion via the calcium sensing receptor (CaSR), antibodies to sclerostin and Dkk1 are currently in clinical trials and may prove to be even more efficient at increasing bone mass, possibly independent of bone turnover. Each of these anabolic approaches has its own limitations and safety issues, but offer good prospects for the anabolic therapy of osteoporosis.[br][br]Nothing to Disclose: REB 2012-06-23T10:30:00 Theater A 2012-06-23T00:00:00 1899-12-30T10:30:00 2506 26 45 S9-3 T03-S04 Saturday 44 2012


2801 ENDO12L_S-LB1-1 LATE-BREAKING SYMPOSIA: CLINICAL - Results of the Origin Trial (9:30 AM - 11:00 AM) To be determined TBD To be determined TBD, TBD Disclosure Incomplete: Author to be determined 2012-06-23T09:30:00 Grand Ballroom AB 2012-06-23T00:00:00 1899-12-30T09:30:00 6264 27 46 S-LB1-1 S-LB1 Saturday 46 2012


2802 ENDO12L_S-LB1-2 LATE-BREAKING SYMPOSIA: CLINICAL - Results of the Origin Trial (9:30 AM - 11:00 AM) To be determined TBD To be determined TBD, TBD Disclosure Incomplete: Author to be determined 2012-06-23T10:00:00 Grand Ballroom AB 2012-06-23T00:00:00 1899-12-30T10:00:00 6265 27 47 S-LB1-2 S-LB1 Saturday 47 2012


2803 ENDO12L_S-LB1-3 LATE-BREAKING SYMPOSIA: CLINICAL - Results of the Origin Trial (9:30 AM - 11:00 AM) To be determined TBD To be determined TBD, TBD Disclosure Incomplete: Author to be determined 2012-06-23T10:30:00 Grand Ballroom AB 2012-06-23T00:00:00 1899-12-30T10:30:00 6266 27 48 S-LB1-3 S-LB1 Saturday 48 2012


2804 ENDO12L_MC1-1 MASTER CLINICIAN: CLINICAL - Difficult Type 1 Diabetes Cases (11:15 AM - 12:15 PM) Difficult Type 1 Diabetes Cases John (Jack) L Leahy University of Vermont College of Medicine, Colchester, VT Session supported by: Lilly USA, LLC[br][br]Disclosure Incomplete: JLL 2012-06-23T11:15:00 Grand Ballroom AB 2012-06-23T00:00:00 1899-12-30T11:15:00 6108 43 110 MC1-1 MC1 Saturday 50 2012


2805 ENDO12L_MC1-2 MASTER CLINICIAN: CLINICAL - Difficult Type 1 Diabetes Cases (11:15 AM - 12:15 PM) Difficult Type 1 Diabetes Cases Irl B Hirsch Univ of Washington Med Ctr, Seattle, WA Session supported by: Lilly USA, LLC[br][br]Disclosure Incomplete: Author to be determined 2012-06-23T11:15:00 Grand Ballroom AB 2012-06-23T00:00:00 1899-12-30T11:15:00 6270 43 111 MC1-2 MC1 Saturday 51 2012


2806 ENDO12L_CDW1 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Setting up a Lab (11:15 AM - 11:45 AM) Setting up a Lab Andrea C Gore University of Texas at Austin, Austin, TX Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: ACG 2012-06-23T11:15:00 Trainee Career Center 2012-06-23T00:00:00 1899-12-30T11:15:00 6216 44 112 CDW1 CDW1 Saturday 52 2012


2807 ENDO12L_SCS1-1 SPECIAL COMPETITION SESSION: CLINICAL - Clinical Research Fellowship Oral Presentations in Women’s Health (11:15 AM - 12:45 PM) To be determined Cevon M Rariy Brigham [amp] Williams Hospital, Boston, MA Session supported by: Pfizer, Inc[br][br]Disclosure Incomplete: Author to be determined 2012-06-23T11:15:00 Room 310 2012-06-23T00:00:00 1899-12-30T11:15:00 6277 45 113 SCS1-1 SCS1 Saturday 55 2012


2808 ENDO12L_SCS1-2 SPECIAL COMPETITION SESSION: CLINICAL - Clinical Research Fellowship Oral Presentations in Women’s Health (11:15 AM - 12:45 PM) To be determined Alexander T Faje Massachusetts General Hospital General Hospital General Hospital, Boston, MA Session supported by: Pfizer, Inc[br][br]Disclosure Incomplete: Author to be determined 2012-06-23T11:30:00 Room 310 2012-06-23T00:00:00 1899-12-30T11:30:00 6278 45 114 SCS1-2 SCS1 Saturday 56 2012


2809 ENDO12L_SCS1-3 SPECIAL COMPETITION SESSION: CLINICAL - Clinical Research Fellowship Oral Presentations in Women’s Health (11:15 AM - 12:45 PM) To be determined Grace Huang Boston Medical Center, Boston, MA Session supported by: Pfizer, Inc[br][br]Disclosure Incomplete: Author to be determined 2012-06-23T11:45:00 Room 310 2012-06-23T00:00:00 1899-12-30T11:45:00 6279 45 115 SCS1-3 SCS1 Saturday 57 2012


2810 ENDO12L_SCS1-4 SPECIAL COMPETITION SESSION: CLINICAL - Clinical Research Fellowship Oral Presentations in Women’s Health (11:15 AM - 12:45 PM) To be determined Lauren W Roth University of Colorado, Aurora, CO Session supported by: Pfizer, Inc[br][br]Disclosure Incomplete: Author to be determined 2012-06-23T12:00:00 Room 310 2012-06-23T00:00:00 1899-12-30T12:00:00 6280 45 116 SCS1-4 SCS1 Saturday 58 2012


2811 ENDO12L_SCS1-5 SPECIAL COMPETITION SESSION: CLINICAL - Clinical Research Fellowship Oral Presentations in Women’s Health (11:15 AM - 12:45 PM) To be determined Carolina C Di Blasi Seattle Children's Hospital, Seattle, WA Session supported by: Pfizer, Inc[br][br]Disclosure Incomplete: Author to be determined 2012-06-23T12:15:00 Room 310 2012-06-23T00:00:00 1899-12-30T12:15:00 6281 45 117 SCS1-5 SCS1 Saturday 59 2012


2812 ENDO12L_SCS1-6 SPECIAL COMPETITION SESSION: CLINICAL - Clinical Research Fellowship Oral Presentations in Women’s Health (11:15 AM - 12:45 PM) To be determined Pornpoj Pramyothin Boston University School of Medicine, Boston, MA Session supported by: Pfizer, Inc[br][br]Disclosure Incomplete: Author to be determined 2012-06-23T12:30:00 Room 310 2012-06-23T00:00:00 1899-12-30T12:30:00 6282 45 118 SCS1-6 SCS1 Saturday 60 2012


2813 ENDO12L_CDW2 CAREER DEVELOPMENT WORKSHOP: CLINICAL - Setting up a Clinical Practice (12:00 PM - 12:30 PM) Setting up a Clinical Practice Elliot G Levy University of Miami Miller School of Medicine, Miami Beach, FL Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: EGL 2012-06-23T12:00:00 Trainee Career Center 2012-06-23T00:00:00 1899-12-30T12:00:00 6253 46 119 CDW2 CDW8 Saturday 61 2012


2815 ENDO12L_W0-1 WORKSHOP: CLINICAL - MOC Made Easy: Renewing Your Certification (2:45 PM - 3:30 PM) MOC Made Easy: Renewing Your Certification Graham T McMahon Brigham and Women[apos]s Hosp, Boston, MA Disclosure Incomplete: Author to be determined 2012-06-23T14:45:00 Theater C 2012-06-23T00:00:00 1899-12-30T14:45:00 6273 89 867 W0-1 W0 Saturday 63 2012


2816 ENDO12L_CMF2-1 CASE MANAGEMENT FORUM: CLINICAL - Cushing[apos]s Disease: Dialogue between an Endocrinologist [amp] a Neurosurgeon (2:45 PM - 3:30 PM) Cushing[apos]s Disease: Dialogue between a Surgeon [amp] an Endocrinologist Specializing in Pituitary Disease James W Findling Medical College of Wisconsin, Menomonee Falls, WI Nothing to Disclose: JWF 2012-06-23T14:45:00 Theater B 2012-06-23T00:00:00 1899-12-30T14:45:00 6260 78 854 CMF2-1 CMF-A1 Saturday 64 2012


2817 ENDO12L_CMF2-2 CASE MANAGEMENT FORUM: CLINICAL - Cushing[apos]s Disease: Dialogue between an Endocrinologist [amp] a Neurosurgeon (2:45 PM - 3:30 PM) Cushing[apos]s Disease: Dialogue between a Surgeon [amp] an Endocrinologist Specializing in Pituitary Disease Brooke Swearingen Massachusetts General Hospital, Boston, MA Disclosure Incomplete: BS 2012-06-23T14:45:00 Theater B 2012-06-23T00:00:00 1899-12-30T14:45:00 6250 78 855 CMF2-2 CMF-A1 Saturday 65 2012


2818 ENDO12L_CMF3-1 CASE MANAGEMENT FORUM: CLINICAL - Diagnosis [amp] Management of Male Hypogonadism in the Older Man (2:45 PM - 3:30 PM) Diagnosis [amp] Management of Male Hypogonadism in the Older Man Frances J Hayes St Vincent[apos]s University Hospital, Dublin, Ireland Disclosure Incomplete: FJH 2012-06-23T14:45:00 Theater A 2012-06-23T00:00:00 1899-12-30T14:45:00 6168 79 856 CMF3-1 CMF-MR1 Saturday 66 2012


2819 ENDO12L_CMF3-2 CASE MANAGEMENT FORUM: CLINICAL - Diagnosis [amp] Management of Male Hypogonadism in the Older Man (2:45 PM - 3:30 PM) Diagnosis [amp] Management of Male Hypogonadism in the Older Man Alvin M Matsumoto VA Puget Sound Health Care System, Seattle, WA Disclosures: AMM: Principal Investigator, Abbott Laboratories, GlaxoSmithKline, Ascend; Consultant, Novartis Pharmaceuticals. 2012-06-23T14:45:00 Theater A 2012-06-23T00:00:00 1899-12-30T14:45:00 6172 79 857 CMF3-2 CMF-MR1 Saturday 67 2012


2820 ENDO12L_M6 MEET-THE-PROFESSOR: CLINICAL - Challenging Multiple Endocrine Neoplasia Cases (2:45 PM - 3:30 PM) Challenging Multiple Endocrine Neoplasia Cases Robert F Gagel University of Texas MD Anderson Cancer Center, Houston, TX Nothing to Disclose: RFG 2012-06-23T14:45:00 Room 351 2012-06-23T00:00:00 1899-12-30T14:45:00 6210 80 858 M6 M1 Saturday 68 2012


2821 ENDO12L_M7 MEET-THE-PROFESSOR: CLINICAL - Chronic Kidney Disease [amp] Bone (2:45 PM - 3:30 PM) Chronic Kidney Disease [amp] Bone Mark Stuart Cooper University of Birmingham, Birmingham, UK Nothing to Disclose: MSC 2012-06-23T14:45:00 Room 371 2012-06-23T00:00:00 1899-12-30T14:45:00 6144 81 859 M7 B7 Saturday 69 2012


2822 ENDO12L_M8 MEET-THE-PROFESSOR: CLINICAL - Dietary Issues in Ethnic Groups (2:45 PM - 3:30 PM) Dietary Issues in Ethnic Groups Abhimanyu Garg University Texas Southwest Medical Center, Dallas, TX Nothing to Disclose: AG 2012-06-23T14:45:00 Room 362 2012-06-23T00:00:00 1899-12-30T14:45:00 6181 82 860 M8 D9 Saturday 70 2012


2823 ENDO12L_M9 MEET-THE-PROFESSOR: CLINICAL - Difficult Thyroid Function Tests (2:45 PM - 3:30 PM) Difficult Thyroid Function Tests Douglas S Ross Massachusetts General Hospital, Waban, MA Disclosures: DSR: Speaker, Genzyme Corporation; Researcher, Genzyme Corporation. 2012-06-23T14:45:00 Room 332 2012-06-23T00:00:00 1899-12-30T14:45:00 6161 83 861 M9 TH4 Saturday 71 2012


2824 ENDO12L_M10 MEET-THE-PROFESSOR: CLINICAL - Hyponatremia (2:45 PM - 3:30 PM) Hyponatremia Joseph G Verbalis Georgetown University, Washington, DC Disclosures: JGV: Consultant, Astellas, Cardiokine, Otsuka, Sanofi-Aventis. 2012-06-23T14:45:00 Room 361 ABDE 2012-06-23T00:00:00 1899-12-30T14:45:00 6166 84 862 M10 PIT3 Saturday 72 2012


2825 ENDO12L_M11 MEET-THE-PROFESSOR: CLINICAL - Insulin Pumps (2:45 PM - 3:30 PM) Insulin Pumps Howard Wolpert Joslin Diabetes Center, Boston, MA Session supported by: Lilly USA, LLC and Medtronic Diabetes[br][br]Disclosure Incomplete: HW 2012-06-23T14:45:00 Room 342 ABDE 2012-06-23T00:00:00 1899-12-30T14:45:00 6249 85 863 M11 D4 Saturday 73 2012


2826 ENDO12L_M13 MEET-THE-PROFESSOR: CLINICAL - Non-Cancer Thyroid [apos]Hot Topics[apos] in Children [amp] Adolescents (2:45 PM - 3:30 PM) Non-Cancer Thyroid [apos]Hot Topics[apos] in Children [amp] Adolescents Stephen H LaFranchi Oregon Health Sciences University, Portland, OR Nothing to Disclose: SHL 2012-06-23T14:45:00 Room 310 2012-06-23T00:00:00 1899-12-30T14:45:00 6167 87 865 M13 PED5 Saturday 74 2012


2827 ENDO12L_M14 MEET-THE-PROFESSOR: CLINICAL - Non-PTH Hypercalcemia (2:45 PM - 3:30 PM) Non-PTH Hypercalcemia Gregory A Clines University of Alabama Birmingham, Birmingham, AL Nothing to Disclose: GAC 2012-06-23T14:45:00 Room 320 2012-06-23T00:00:00 1899-12-30T14:45:00 6141 90 868 M14 B2 Saturday 75 2012


2828 ENDO12L_M15 MEET-THE-PROFESSOR: CLINICAL - Premature Ovarian Failure ( Insufficiency) (2:45 PM - 3:30 PM) Premature Ovarian Insufficiency Nanette F Santoro University of Colorado School of Medicine, Aurora, CO Nothing to Disclose: NFS 2012-06-23T14:45:00 Room 352 DEF 2012-06-23T00:00:00 1899-12-30T14:45:00 6151 88 866 M15 FR5 Saturday 76 2012


2829 ENDO12L_PS1-1 PRESIDENTIAL SYMPOSIUM SESSION: TRANSLATIONAL - Endocrine Disruptors with NIEHS Director Linda Birnbaum (3:45 PM - 5:15 PM) How Environmental Chemicals Target Multiple Endocrine Pathways: Perspectives from the NIEHS Director [mdash] Linda Birnbaum Linda S Birnbaum National Institute of Environmental Health Sciences/NIH, Research Triangle Park, NC The endocrine system is a highly organized system of glands and hormones that regulates vital functions such as growth, response to stress, sexual development and behavior, production and utilization of insulin, metabolism, intelligence and behavior, and the ability to reproduce. It is now clear that this system can be perturbed by environmental chemicals that were designed for one effect but have been shown to interfere with endocrine signaling.[br]To date, there are over 800 of these so-called endocrine disrupting chemicals (EDCs) which include pharmaceuticals, dioxin and dioxin-like compounds, polychlorinated biphenyls, DDT and other pesticides, and components of plastics such as bisphenol A (BPA), phthalates, flame retardants and solvents. EDCs are found in many everyday products--including plastic bottles, metal food cans, detergents, foam and fabric, food additives, toys, cosmetics, and household products.[br]EDCs were originally thought to exert their actions solely through nuclear hormone receptors, including receptors for estrogen, androgen, progesterone, thyroid hormones, and retinoids. However, recent evidence shows that the mechanisms by which EDCs, as well as endogenous hormones, act are much broader than originally recognized. Studies have shown that in addition to altering nuclear receptor signaling, EDCs are capable of acting through membrane receptors, nonsteroid receptors, transcriptional coactivators and repressors, that can all converge upon endocrine, reproductive, metabolic, and neurological systems, among others. Many EDCs can perturb multiple endocrine pathways, and effects are context (time and tissue) dependent.[br]Chemical exposures during early life stages can disrupt normal patterns of development and thus dramatically alter disease susceptibility later in life. EDC exposures during developmental windows have been shown to affect endocrine signaling pathways that can lead to endocrine cancers, obesity, diabetes, infertility, premature puberty, learning disabilities, cognitive and brain development problems, sexual development problems including feminizing of males or masculine effects on females, etc. In short, any system in the body controlled by hormones, can be derailed by EDCs.[br][br]Nothing to Disclose: LSB 2012-06-23T15:45:00 Room 332 2012-06-23T00:00:00 1899-12-30T15:45:00 2475 92 869 PS1-1 SSS01 Saturday 78 2012


2830 ENDO12L_PS1-2 PRESIDENTIAL SYMPOSIUM SESSION: TRANSLATIONAL - Endocrine Disruptors with NIEHS Director Linda Birnbaum (3:45 PM - 5:15 PM) Novel Perspectives on Hormone-Receptor Interactions [amp] Implications for Environmental Chemicals Donald Patrick McDonnell Duke University School of Medicine, Durham, NC Nothing to Disclose: DPM 2012-06-23T16:15:00 Room 332 2012-06-23T00:00:00 1899-12-30T16:15:00 6204 92 870 PS1-2 SSS01 Saturday 79 2012


2831 ENDO12L_PS1-3 PRESIDENTIAL SYMPOSIUM SESSION: TRANSLATIONAL - Endocrine Disruptors with NIEHS Director Linda Birnbaum (3:45 PM - 5:15 PM) Children[apos]s Vulnerability to Environmental Chemicals Philip J Landrigan Mt Sinai School of Medicine, New York, NY Disclosure Incomplete: PJL 2012-06-23T16:45:00 Room 332 2012-06-23T00:00:00 1899-12-30T16:45:00 6171 92 871 PS1-3 SSS01 Saturday 80 2012


2832 ENDO12L_S10-1 SYMPOSIUM SESSION: TRANSLATIONAL - Advances in Adrenal Tumors (3:45 PM - 5:15 PM) Wnt Signaling in Adrenal Tumorigenesis Isabelle Bourdeau CHUM, St Lambert, Canada In the past few years, the key role of Wnt signaling in adrenal gland development and tumors has been better understood. We will review canonical Wnt signaling alterations in adrenocortical tumorigenesis. In the absence of a Wnt signal, [beta]-catenin is captured by APC and axin, facilitating its phosphorylation by casein kinase 1 [alpha] (CK1[alpha]) and glycogen synthase kinase 3[beta] (GSK3[beta]). Phosphorylated [beta]-catenin is then available for proteosomal degradation. In contrast, Wnt activation inhibits GSK-3[beta]; consequently, [beta]-catenin accumulates and forms complexes with Tcf/Lef proteins in the nucleus, stimulating the expression of various Wnt target genes. [beta]-catenin gene ([italic]CTNNB1[/italic]) mutations are found in more than 20% of sporadic adrenocortical tumors. Most [beta]-catenin mutations occur at critical serine and tyrosine residues of exon3 of [italic]CTNNB1[/italic] gene, which leads to cytoplasmic and nuclear accumulation of the [beta]-catenin protein. Somatic [beta]-catenin mutations were also identified in large PPNAD nodules. Cytoplasmic and nuclear [beta]-catenin protein accumulation is seen in a higher percentage of tumors than [italic]CTNNB1[/italic] mutations, indicating that other factors of the Wnt/[beta]-catenin signaling pathway may evoke [beta]-catenin accumulation in adrenocortical tumors. The [italic]APC[/italic] gene is among the candidates that were studied. Mostly silent somatic mutation and polymorphisms were found in [italic]APC[/italic] in ACC. Recently, we identified genetic alterations in the [italic]AXIN2[/italic] gene in 7% of AA and 17% of ACC and in the human ACC H295R cell line in association with [beta]-catenin protein accumulation. In a cohort of ACC, [beta]-catenin nuclear staining and [italic]CTNNB1[/italic] mutations were linked with decreased overall and disease-free survival.[br][italic]In vitro[/italic] studies of the H295R ACC cell line demonstrated that activating [beta]-catenin mutation leads to efficient T-cell factor dependent transcription and that its inhibition with Tcf/[beta]-catenin antagonist decreased cell proliferation. Transgenic mice expressing a constitutively active [beta]-catenin in their adrenal cortex developed adrenal hyplasia and dysplasia. Over a 17-month period, transgenic adrenals developed malignant characteristics and primary hyperaldosteronism. Altogether, these findings suggest that therapeutic modulation of the Wnt pathway may be an attractive possibility for treating patients with adrenocortical carcinoma.[br][br]Sources of Research Support: Fonds de la Recherche en Sant[eacute] du Qu[eacute]bec (Grant FRSQ-6519/5360) and The Cancer Research Society.[br][br]Nothing to Disclose: IB 2012-06-23T15:45:00 Theater A 2012-06-23T00:00:00 1899-12-30T15:45:00 2540 93 872 S10-1 T01-S01 Saturday 82 2012


2833 ENDO12L_S10-2 SYMPOSIUM SESSION: TRANSLATIONAL - Advances in Adrenal Tumors (3:45 PM - 5:15 PM) Pathology Algorithms for ACC Thomas J Giordano University of Michigan Health System, Ann Arbor, MI Disclosure Incomplete: TJG 2012-06-23T16:15:00 Theater A 2012-06-23T00:00:00 1899-12-30T16:15:00 6002 93 873 S10-2 T01-S01 Saturday 83 2012


2834 ENDO12L_S10-3 SYMPOSIUM SESSION: TRANSLATIONAL - Advances in Adrenal Tumors (3:45 PM - 5:15 PM) Results from New Trials for Andrenocortical Cancer Bruno Allolio University of Wuerzburg, Wuerzburg, Germany In 2012 the final results of the first randomized trial for advanced adrenocortical cancer (ACC) comparing etoposide, doxorubicin, and cisplatin plus mitotane (EDP-M) with streptozotocin plus mitotane (Sz-M) will become available. As compared to Sz-M in first-line (N=153), patients assigned to EDP-M first (N=151) had a significantly higher response rate (23.2% vs. 9.2%, P[lt]0.001), a longer median progression-free survival (5.0 vs. 2.1 months, HR 0.55, 95%CI 0.43[ndash]0.69, P[lt]0.001), while overall survival was not significantly different (14.8 vs. 12.0 months, HR 0.79, 95%CI 0.61[ndash]1.02; P=0.071), as most patients switched to the alternate regimen in case of progressive disease. Patients not treated with the alternate therapy in second-line, had better overall survival with first-line EDP-M (17.1 months, N=67) than Sz-M (4.7 months; N=52). Serious adverse event rates did not differ significantly between treatments. These results indicate that EDP-M should become the standard of care in advanced ACC not responding to mitotane. For patients progressing after EDP-M treatment with gemcitabine plus metronomic fluoropyrimidines is well tolerated and promising with [gt]40% of patients not progressing after 4 months. In contrast, neither inhibition of EGF receptor nor targeting VEGF resulted in a benefit in advanced ACC. Overexpression of IGF-2 in [gt]90% of ACCs has led to trials investigating inhibition of IGF-1R signaling with monoclonal antibodies (figitumomab, cixutumomab) or small -molecule inhibitors (OSI-906). Early trials have shown promising activity, but results of a more informative phase III trial using OSI-906 are still pending. Combination of IGF1-R blockade with other targeted therapies (e.g. temsirolimus) may lead to improved activity. Recently it has become evident that mitotane by strongly inducing Cyp3A4 impacts on the efficacy of tyrosine kinase inhibitors, as standard doses may lead to insufficient drug levels. Accordingly, in a phase II trial with sunitinib (n=35) the rate of stable disease was higher in patients without concomitant mitotane suggesting that drug monitoring may be essential in trials using targeted treatments in combination with mitotane. As an alternative option, a first series of patients (n=11) with advanced ACC was treated with 131I-Iodometomidate. The results indicate promising activity in tumors expressing high levels of Cyp11B enzymes.[br][br]Nothing to Disclose: BA 2012-06-23T16:45:00 Theater A 2012-06-23T00:00:00 1899-12-30T16:45:00 2425 93 874 S10-3 T01-S01 Saturday 84 2012


2835 ENDO12L_S11-1 SYMPOSIUM SESSION: CLINICAL - Androgens [amp] Prostate Cancer: 75 Years after Huggins [amp] Hodges (3:45 PM - 5:15 PM) Chemoprevention of Prostate Cancer by 5 Alpha-Reductase Inhibitors (5ARIs): Emergent Data [amp] Controversies Roger S Rittmaster Dalhousie University , Camden, ME Two placebo-controlled studies have demonstrated the ability of 5ARIs to decrease the risk of being diagnosed with prostate cancer. In the Prostate Cancer Prevention Trial (PCPT), a 7-year trial involving over 18,000 men, finasteride reduced the risk of prostate cancer by 25% in men over age 55 with a PSA level [le] 3.0 at baseline.(1) In the REDUCE trial, a 4-year trial involving over 8,000 men, dutasteride reduced the risk of prostate cancer by 23% in men over age 50 who had a baseline PSA [ge] 2.5 and a negative prostate biopsy done for suspicion of prostate cancer.(2) The prostate cancer risk reduction in both studies was due to a reduction in low grade cancer (Gleason score [le] 6), and in both studies there was an increase in the detection of high grade cancer (Gleason 7-10 in the PCPT and 8-10 in REDUCE). Whether finasteride and dutasteride increase the growth of high grade cancers or enhance the detection of such cancers has been widely debated. The difference is critical: enhanced detection of potentially lethal cancers would be a benefit. Design features in both studies could have led to an enhanced diagnosis of high grade cancers in the 5ARI group. In both studies the rates of clinically advanced prostate cancer were similar in the two study arms.[br]It has been argued that the cancers prevented by 5ARIs are [ldquo]insignificant[rdquo], based on pathological criteria. However, up to 90% of low grade prostate cancers are aggressively treated, and from the patient[apos]s perspective, there are few cancers that are clinically insignificant. Hence, one benefit of prostate cancer chemoprevention is a reduction in the morbidity and mortality of aggressive treatment of indolent prostate cancers. This would be a dubious benefit if it were not for PSA screening, which has led to the overdiagnosis and overtreatment of prostate cancer. Nevertheless, it is a difficult argument to propose a medical treatment to help overcome the limitations of a poor screening test.[br]One mechanism by which 5ARIs could reduce the risk of prostate cancer is by shrinking existing cancers, low grade more than high grade. In the recently-reported REDEEM study, in which dutasteride or placebo was given to 302 men with low grade prostate cancer being followed by active surveillance, the dutasteride group had a lower rate of prostate cancer progression and less likelihood of prostate cancer being detected on follow-up biopsy.(3)[br][br](1) Thompson IM et al., New Engl J Med 2003; 349:215. (2) Andriole GL et al., New Engl J Med 2010; 362:1192. (3) Fleshner NE et al., Lancet 2012. Epub 2012/01/27.[br][br]Disclosures: RSR: Consultant, Abbott Laboratories, GlaxoSmithKline; Ad Hoc Consultant, Sanofi-Aventis. 2012-06-23T15:45:00 Grand Ballroom AB 2012-06-23T00:00:00 1899-12-30T15:45:00 2388 94 875 S11-1 T05-S04 Saturday 86 2012


2836 ENDO12L_S11-2 SYMPOSIUM SESSION: CLINICAL - Androgens [amp] Prostate Cancer: 75 Years after Huggins [amp] Hodges (3:45 PM - 5:15 PM) Castration-Resistant Prostate Cancer: Mechanisms [amp] Management Howard Scher Memorial Sloan Kettering Cancer Center, New York, NY Tumor regrowth despite castrate levels of testosterone (T) results in part from a series of oncogenic changes in the androgen receptor (AR) and the AR signaling axis including upregulation of the androgen biosynthetic machinery leading to increased intratumoral androgens, AR overexpression, AR mutations and ligand-independent activation by growth factor signaling pathways. These findings have been validated in phase 3 trials of the CYP17 inhibitor abiraterone acetate (AA) and the androgen signaling inhibitor MDV3100 in patients with docetaxel-treated CRPC. In COU-301, AA + prednisone increased the median OS from 10.9 to 14.8 months relative to placebo (HR=0.65; p[lt]0.001)(1) while in AFFIRM, MDV3100 was shown to prolong median OS from 13.6 to 18.4 months (HR=0.63; p[lt]0.0001)(2). The results establish definitively that tumors progressing despite castrate levels of T are not hormone-refractory. Regarding tumors as refractory to additional hormone therapy may deny patients effective life prolonging treatment. With the development of additional agents targeting the AR signaling cascade, designing trials to determine how to maximize their benefit alone or in combination and in what sequence is increasingly important. The CYP17 inhibitors TAK-700 and TOK-001 and the AR signaling inhibitor ARN-509, shown to have greater efficacy than MDV3100 in xenograft models(3) are also in phase 3 development. Critical for their advancement, particularly as it pertains to drug approvals, is the co-development of efficacy-response surrogate biomarkers for survival. A qualification effort in collaboration with the FDA using circulating tumor cell biomarkers is ongoing.[br]A similar pattern of response to these agents exists: dramatic and durable responses in some, an initial response followed by progression in others, while a proportion are resistant de novo. Key is identifying predictive biomarkers of sensitivity, establishing analytically valid assays for them, and developing trials in sequence to generate evidence to qualify them for specific contexts of use. Early work focused on the presence or absence of TMPRSS2:ERG fusions, which place an ETS transcription factor under androgen regulation(4). Building on the observation in laboratory models showing the reciprocal negative feedback between AR and PI3K signaling(5), multiple trials have been developed sequencing AA with PI3K/Akt/mTOR inhibitors. Potential predictive biomarkers of sensitivity are embedded into these trials.[br][br](1) Fizazi K, et al., Eur J Cancer 2011; 47:S483. (2) Scher HI, et al., J Clin Oncol 2012; 30:LBA1. (3) Clegg NJ, et al., Cancer Res 2012. (4) Danila DC, et al., Eur Urol 2011; 60:897. (5) Carver BS, et al., Cancer Cell 2011; 19:575.[br][br]Disclosures: HS: Consultant, Amgen, Bristol-Myers Squibb, Sanofi-Aventis, Dendreon, Endo Pharmaceuticals Inc., Ortho Biotech Oncology Research & Development, Millennium, Novartis Pharmaceuticals, Senior Scientific LLC; Principal Investigator, Medivation, Ortho Biotech Oncology Research & Development, Aragon, Bristol-Myers Squibb, Exelixis, Inc., Veridex. 2012-06-23T16:15:00 Grand Ballroom AB 2012-06-23T00:00:00 1899-12-30T16:15:00 2520 94 876 S11-2 T05-S04 Saturday 87 2012


2837 ENDO12L_S11-3 SYMPOSIUM SESSION: CLINICAL - Androgens [amp] Prostate Cancer: 75 Years after Huggins [amp] Hodges (3:45 PM - 5:15 PM) Management of Side Effects of Androgen Deprivation Therapy Jeffrey D Zajac University of Melbourne, Heidelberg, Australia Androgen deprivation therapy (ADT) has major beneficial effects on patients with prostate cancer. It is widely used for palliation in metastatic disease but also in localized disease and for biochemical recurrence. ADT is now the commonest diagnosed cause of severe hypogonadism in adult men. However this very effective therapy causes substantial metabolic and other side effects (1,2). The commonest cause of death in men with prostate cancer is cardiovascular disease and ADT worsens the risk profile. Osteoporosis, changes in glucose metabolism and changes in mood and sexual function are common.[br]ADT results in an increase in fat mass of 5-10% at 6 months, in particular an increase in abdominal fat mass, as well as a 3% loss of muscle mass. These metabolically unfavorable changes in body composition promote insulin resistance and increase the risk of diabetes. These and associated changes in the lipid profile worsen the cardiovascular risk. Bone loss is identifiable at 6 months with changes in microarchitecture. The rate of loss increases 5-10 fold and there is an increase in fracture risk. Substantial loss of muscle mass increases the risk of falls and fractures. A 15% drop in hemoglobin occurs but it is not clear if this translates into morbidity.[br]ADT has profound effects on sexual function. Less than 20% of men on ADT have any sexual interest. In one study erectile dysfunction rose from 38% to 74%. Hot flushes are common and changes in body image, mood and cognitive function have been documented. Many of the studies describing these effects are small and short term with no adequate controls. Most rely on community-based controls rather than prostate cancer patients not receiving ADT.[br]Management of metabolic problems is well characterized in the general population and although evidence in prostate cancer patients is lacking it is prudent to treat these issues in such patients in a similar fashion. Evidence exists for the management of the bone loss with antiresorptive agents.[br]The side effects of ADT are manageable and where there is evidence of survival benefit ADT is indicated. However given that patients with prostate cancer are already at significant risk of osteoporotic fracture, diabetes and cardiovascular disease before they start ADT particular attention should be given to these issues when they do start. Where the evidence of benefit is lacking due consideration should be given to the side effects in any decision to commence ADT.[br][br](1) Grossmann M, Hamilton EJ, Gilfillan C, Bolton D, Joon DL, Zajac JD. Bone and metabolic health in patients with non-metastatic prostate cancer who are receiving androgen deprivation therapy. Med J Aust 194:301-306, 2011. (2) Grossmann M, Zajac JD. Management of side effects of androgen deprivation therapy. Endocrinol Metab Clin North Am 40:3; 655-71,2011.[br][br]Sources of Research Support: NHMRC.[br][br]Nothing to Disclose: JDZ 2012-06-23T16:45:00 Grand Ballroom AB 2012-06-23T00:00:00 1899-12-30T16:45:00 2483 94 877 S11-3 T05-S04 Saturday 88 2012


2854 ENDO12L_S17-2 SYMPOSIUM SESSION: CLINICAL - Pediatric Implications of Reproductive Endocrine Technologies (3:45 PM - 5:15 PM) The Female Adolescent [amp] Young Adult at Risk for Ovarian Failure: Fertility Preservation [amp] Assisted Reroduction Options in 2012 Kutluk Oktay New York Medical College, Valhalla, NY Preservation of ovarian function and future fertility are largely neglected in children who are faced with conditions that can result in premature ovarian failure. Hence compared to adults, children experience health care disparity in the field of fertility preservation. In addition to chemotherapy and radiotherapy, genetic conditions such as FMR1 premutations or Turner[apos]s syndrome, or metabolic disorders such as galactosemia can result in premature ovarian failure in adolescents or young adults. In the recent decade, numerous options have emerged for preserving fertility in the presence of high risk factors for early loss of ovarian function and fertility in young females. These include ovarian cryoreservation and transplantation, as well as oocyte cryopreservation (1, 2, 3, 4). There is also the promise of ceramide-induced death pathway inhibitors in preventing chemotherapy-induced germ cell death (5) When already faced with premature ovarian failure, future fertility options include oocyte donation, surrogacy (gestational carriers) and adoption. Although recent research suggested there maybe ovarian germ stem cells in adult ovaries, the practical applications of these new findings is unknown. Our current research also indicates that oocytes have the ability to repair DNA damage that occurs with genotoxic stress which may bring the theoretical possibility of oocyte [quot]rejuvenation[quot] (6). Given the current state of the field of fertility preservation and assisted reproduction, young females should be properly counseled and immediately referred to appropriate fertility specialists when faced with the possibility of early loss of gonadal function.Those who have not yet experienced premature ovarian failure but at risk for early menopause can be identified early by serum AMH levels and antral follicle counts (2,3) and can be offered fertility preservation or other assisted reproduction techniques to preserve or restore fertility.(7)[br][br](1)Rodriguez-Wallberg & Oktay, Fertility preservation medicine: options for young adults and children with cancer. J Pediatr Hematol Oncol. 2010 Jul;32(5):390-6. (2)Oktay K and Karlikaya G. Ovarian function after transplantation of frozen, banked autologous ovarian tissue. N Engl J Med. 2000 Jun 22;342(25):1919. (3)Purushothaman R, Lazareva O, Oktay K, Ten S.Markers of ovarian reserve in young girls with Turner[apos]s syndrome. Fertil Steril. 2010 Sep;94(4):1557-9. Epub 2010 Jan 25. (4)Oktay K, Rodriguez-Wallberg KA, Sahin G. Fertility preservation by ovarian stimulation and oocyte cryopreservation in a 14-year-old adolescent with Turner syndrome mosaicism and impending premature ovarian failure. Fertil Steril. 2010 Jul;94(2):753.e15-9. (5) Soleimani R, Heytens E, and Oktay K. Prevention of chemotherapy-induced primordial follicle death in human ovarian xenografts. Abstract. 2010 Annual Meeting of American Society of Reproductive Medicine. (6)Soleimani R, Heytens E, Darzynkiewicz Z, Oktay K.Mechanisms of chemotherapy-induced human ovarian aging: double strand DNA breaks and microvascular compromise. Aging (Albany NY). 2011 Aug;3(8):782-93. (7)www.fertilitypreservation.org.[br][br]Sources of Research Support: The National Institute of Health NICHD grants, RO1HD053112 and R21HD061259, awarded to KO.[br][br]Nothing to Disclose: KO 2012-06-23T16:15:00 Room 310 2012-06-23T00:00:00 1899-12-30T16:15:00 2543 100 894 S17-2 T07-S04 Saturday 111 2012


2855 ENDO12L_S17-3 SYMPOSIUM SESSION: CLINICAL - Pediatric Implications of Reproductive Endocrine Technologies (3:45 PM - 5:15 PM) Pre-Implantation Genetic Diagnosis for Families Living with Serious Hereditary Endocrine Diseases Mark R Hughes Genesis Genetics Institute, Detroit, MI Nothing to Disclose: MRH 2012-06-23T16:45:00 Room 310 2012-06-23T00:00:00 1899-12-30T16:45:00 6196 100 895 S17-3 T07-S04 Saturday 112 2012


2856 ENDO12L_L2-1 PLENARY: TRANSLATIONAL - Clinical Investigator Award Lecture: Cellular Mechanisms of Insulin Resistance: Implications for Obesity, Lipodystrophy, Type 2 Diabetes [amp] the Metabolic Syndrome (5:30 PM - 6:30 PM) Clinical Investigator Award Lecture: Cellular Mechanisms of Insulin Resistance [mdash] Implications for Obesity, Lipodystrophy, Type 2 Diabetes [amp] the Metabolic Syndrome Gerald I Shulman Yale University School of Medicine/Howard Hughes Medical Institute, New Haven, CT Insulin resistance is a major factor in the pathogenesis of type 2 diabetes and the metabolic syndrome. Here we will review recent studies, using magnetic resonance spectroscopy, that have implicated ectopic lipid deposition in liver and skeletal muscle as a causal and unifying factor in the pathogenesis of insulin resistance associated with obesity, lipodystrophy, type 2 diabetes and the metabolic syndrome.[br][br](1) Shulman GI. Cellular mechanisms of insulin resistance. J Clin Invest. 2000;106(2):171-6. PMID:10903330, PMC314317. (2) Petersen KF, Oral EA, Dufour S, Befroy D, Ariyan C, Yu C, Cline GW, DePaoli AM, Taylor SI, Gorden P, Shulman GI. Leptin reverses insulin resistance and hepatic steatosis in patients with severe lipodystrophy. J Clin Invest. 2002;109(10):1345-50. PMID:12021250, PMC150981. (3) Petersen KF, Dufour S, Befroy D, Lehrke M, Hendler RE, Shulman GI. Reversal of nonalcoholic hepatic steatosis, hepatic insulin resistance, and hyperglycemia by moderate weight reduction in patients with type 2 diabetes. Diabetes. 2005;54(3):603-8. PMID:15734833, PMC2995496. (4) Samuel VT, Liu ZX, Qu X, Elder BD, Bilz S, Befroy D, Romanelli AJ, Shulman GI. Mechanism of hepatic insulin resistance in non-alcoholic fatty liver disease. J Biol Chem. 2004;279(31):32345-53. PMID:15166226. (5) Samuel VT, Liu ZX, Wang A, Beddow SA, Geisler JG, Kahn M, Zhang XM, Monia BP, Bhanot S, Shulman GI. Inhibition of protein kinase Cepsilon prevents hepatic insulin resistance in nonalcoholic fatty liver disease. J Clin Invest. 2007;117(3):739-45. PMID:17318260, PMC1797607. (6) Erion DM, Shulman GI. Diacylglycerol-mediated insulin resistance. Nat Med. 2010;16(4):400-2. PMID:20376053. (7) Samuel VT, Petersen KF, Shulman GI. Lipid-induced insulin resistance: unravelling the mechanism. Lancet. 2010;375(9733):2267-77. PMID:20609972, PMC2995547. (8) Petersen KF, Dufour S, Hariri A, Nelson-Williams C, Foo JN, Zhang XM, Dziura J, Lifton RP, Shulman GI. Apolipoprotein C3 gene variants in nonalcoholic fatty liver disease. N Engl J Med. 2010;362(12):1082-9. PMID:20335584, PMC2976042. (9) Lee HY, Birkenfeld AL, Jornayvaz FR, Jurczak MJ, Kanda S, Popov V, Frederick DW, Zhang D, Guigni B, Bharadwaj KG, Choi CS, Goldberg IJ, Park JH, Petersen KF, Samuel VT, Shulman GI. Apolipoprotein CIII overexpressing mice are predisposed to diet-induced hepatic steatosis and hepatic insulin resistance. Hepatology;54(5):1650-60. 2011. PMID:21793029, PMC3205235. (10) Jornayvaz FR, Birkenfeld AL, Jurczak MJ, Kanda S, Guigni BA, Jiang DC, Zhang D, Lee HY, Samuel VT, Shulman GI. Hepatic insulin resistance in mice with hepatic overexpression of diacylglycerol acyltransferase 2. Proc Natl Acad Sci U S A. 2011;108(14):5748-52. PMID:21436037, PMC3078388. (11) Kumashiro N, Erion D, Zhang D, Kahn M, Beddow S, Chu X, Still C, Gerhard G, Han X, Dziura J, Petersen K, Samuel V, Shulman GI. Cellular Mechanism of Insulin Resistance in Nonalcoholic Fatty Liver Disease. Proc Natl Acad Sci U S A. 2011;108(39):16381-5. PMID: 21930939, PMC3182681. (12) Samuel, VT and Shulman, GI Integrating Mechanisms for Insulin Resistance: Common Threads and Missing Links. Cell. (in press 2012).[br][br]Sources of Research Support: National Institutes of Health: DK-40936, DK-49230, DK-085638. Howard Hughes Medical Institute.[br][br]Disclosure Incomplete: GIS 2012-06-23T17:30:00 Grand Ballroom AB 2012-06-23T00:00:00 1899-12-30T17:30:00 2433 114 916 L2-1 PL02-1 Saturday 113 2012


2857 ENDO12L_L2-2 PLENARY: TRANSLATIONAL - Transcriptional Control of Adipose Biology [amp] Energy Homeostasis: Toward Novel Therapeutics (5:30 PM - 6:30 PM) Transcriptional Control of Adipose Biology [amp] Energy Homeostasis: Toward Novel Therapeutics Bruce Spiegelman Dana Farber Cancer Institute/Harvard Medical School, Boston, MA Nothing to Disclose: BS 2012-06-23T17:30:00 Grand Ballroom AB 2012-06-23T00:00:00 1899-12-30T17:30:00 6120 115 917 L2-2 PL02-2 Saturday 114 2012


2858 ENDO12L_NS1 ENDOCRINE NURSES SYMPOSIA: CLINICAL -Pancreas [amp] Islet Cell Transplantation & Biologic Insulin Replacement: Where Are We Now [amp] Where Are We Going? (7:30 AM - 8:30 AM) Islet Cell Transplantation: Current Status Alvin C Powers Vanderbilt University Medical Center, Nashville, TN Disclosure Incomplete: ACP 2012-06-24T07:30:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T07:30:00 6230 121 923 NS1 ENS1 Sunday 115 2012


2859 ENDO12L_L3-1 PLENARY: TRANSLATIONAL - Gerald D. Aurbach Award Lecture: Novel Targets for Insulin Signaling in the Heart [amp] Vasculature (8:00 AM - 9:15 AM) Gerald D. Aurbach Award Lecture: Novel Targets for Insulin Signaling in the Heart [amp] Vasculature E Dale Abel University of Utah School of Medicine, Salt Lake City, UT Nothing to Disclose: EDA 2012-06-24T08:00:00 Grand Ballroom AB 2012-06-24T00:00:00 1899-12-30T08:00:00 6121 122 924 L3-1 PL03-1 Sunday 116 2012


2860 ENDO12L_L3-2 PLENARY: TRANSLATIONAL - Causes [amp] Consequences of Obesity: Insights from Genetics (8:00 AM - 9:15 AM) Causes [amp] Consequences of Obesity: Insights from Genetics I Sadaf Farooqi University of Cambridge Metabolic Research Laboratories, Cambridge, UK Disclosure Incomplete: ISF 2012-06-24T08:00:00 Grand Ballroom AB 2012-06-24T00:00:00 1899-12-30T08:00:00 6123 123 925 L3-2 PL03-2 Sunday 117 2012


2861 ENDO12L_NS2 ENDOCRINE NURSES SYMPOSIA: CLINICAL - Atypical Hip Fractures Related to IV Bisphosphonates (4:00 PM - 5:00 PM) Atypical Hip Fractures Relatled to Bisphosphonates Kenneth Mathis The Methodist Hospital, Houston, TX Disclosure Incomplete: KM 2012-06-24T08:30:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T08:30:00 6231 231 1812 NS2 ENS2 Sunday 118 2012


2862 ENDO12L_CMF4-1 CASE MANAGEMENT FORUM: CLINICAL - Endocrine Disease [amp] Bone: Diagnostic [amp] Management Issues (8:30 AM - 9:15 AM) Endocrine Disease [amp] Bone: Diagnostic [amp] Management Issues Lynn Kohlmeier Spokane Osteoporosis, Spokane, WA Nothing to Disclose: LK 2012-06-24T08:30:00 Room 310 2012-06-24T00:00:00 1899-12-30T08:30:00 6143 125 927 CMF4-1 CMF-B1 Sunday 119 2012


2863 ENDO12L_CMF4-2 CASE MANAGEMENT FORUM: CLINICAL - Endocrine Disease [amp] Bone: Diagnostic [amp] Management Issues (8:30 AM - 9:15 AM) Endocrine Disease [amp] Bone: Diagnostic [amp] Management Issues Jan M Bruder University of Texas HSC at San Antonio, San Antonio, TX Nothing to Disclose: JMB 2012-06-24T08:30:00 Room 310 2012-06-24T00:00:00 1899-12-30T08:30:00 6193 125 928 CMF4-2 CMF-B1 Sunday 120 2012


2864 ENDO12L_CMF5-1 CASE MANAGEMENT FORUM: CLINICAL - Primary Hyperaldosteronism (8:30 AM - 9:15 AM) Primary Hyperaldosteronism Robert M Carey University of Virginia Health System, Charlottesville, VA Nothing to Disclose: RMC 2012-06-24T08:30:00 Room 362 2012-06-24T00:00:00 1899-12-30T08:30:00 6173 126 929 CMF5-1 CMF-A2 Sunday 121 2012


2865 ENDO12L_CMF5-2 CASE MANAGEMENT FORUM: CLINICAL - Primary Hyperaldosteronism (8:30 AM - 9:15 AM) Interaction of Aldosterone [amp] Salt on Cardiovascular Remodeling Michael Stowasser University of Queensland School of Medicine, Brisbane, Australia It is now well recognized that aldosterone (aldo) excess in primary aldosteronism (PA) results in cardiovascular (CV) and renal damage and increased CV morbidity independent of its effects on BP. Experimental studies have indicated that the combined effects of aldo excess and high-salt ingestion are necessary for induction of target-organ damage. We have studied the role of salt in determining urinary protein (Uprot) excretion and changes in left ventricular (LV) structure and the relationship between aldo and salt appetite in patients with PA. Subjects (n=24) with aldo-producing adenoma (APA) underwent measurement of 24 h Uprot and UNa before and after unilateral adrenalectomy (ADX - follow-up 15[plusmn]12 months SD). Following ADX, mean clinic systolic BP fell (150[plusmn]18 vs 135[plusmn]15mmHg, p=0.0008), despite reduction in antihypertensive medications, and Uprot decreased (211[plusmn]102 vs 106[plusmn]42mg per day, P[lt]0.0001). There was a positive correlation between Uprot and UNa both before (r=0.55, p=0.006) and after (r=0.51, p=0.011) ADX and changes in UNa independently predicted Uprot reduction (p=0.019). In a case-control study of 21 patients with PA and 21 with essential hypertension (EH) matched for age, gender, duration of hypertension and 24h systolic and diastolic BP, PA patients had significantly greater mean LV wall thicknesses and mass. UNa positively correlated with, and was an independent predictor of, LV wall thicknesses and LV mass in PA but not EH patients. We also compared pre- and post-ADX UNa, in 93 patients with APA collected on the last day (day 4) of fludrocortisone suppression testing (FST), and, in 35 patients, collected out of hospital. Day 4 UNa decreased from 328[plusmn]79 mmol/day pre-operatively to 272[plusmn]69 mmol/day (p[lt]0.0001) postoperatively despite equal salt supplementation. Out of hospital UNa fell from 172[plusmn]57 to 147[plusmn]56 mmol/day (p=0.0183). Interestingly, UNa on day 4 FST fell significantly only in patients whose FST after ADX showed biochemical cure of PA (n=78), but not in those with remaining autonomous aldo production (n=15). The above results indicate that dietary salt modulates aldo-induced renal and cardiac damage in patients with PA. Hence, apart from that afforded by reduction of aldo effects (by ADX or mineralocorticoid receptor blockade), dietary salt restriction should provide additional target-organ protection in patients with PA. This may be easier to achieve in treated patients as aldo reduction appears to reduce salt appetite.[br][br]Sources of Research Support: Irene Hunt Hypertension Research Trust.[br][br]Nothing to Disclose: MS 2012-06-24T08:30:00 Room 362 2012-06-24T00:00:00 1899-12-30T08:30:00 2486 126 930 CMF5-2 CMF-A2 Sunday 122 2012


2866 ENDO12L_M16 MEET-THE-PROFESSOR: CLINICAL - Adult Turner Syndrome (8:30 AM - 9:15 AM) Adult Turner Syndrome Carolyn Ann Bondy NIH - NICHD, Bethesda, MD Disclosure Incomplete: CAB 2012-06-24T08:30:00 Room 361 ABDE 2012-06-24T00:00:00 1899-12-30T08:30:00 6242 127 931 M16 FR3 Sunday 123 2012


2867 ENDO12L_M17 MEET-THE-PROFESSOR: CLINICAL - Causes of the Condition [amp] the Epidemic (8:30 AM - 9:15 AM) Childhood Obesity: Causes Tamara S Hannon Riley Hospital for Children, Indianapolis, IN Nothing to Disclose: TSH 2012-06-24T08:30:00 Room 360 2012-06-24T00:00:00 1899-12-30T08:30:00 6191 128 932 M17 PED1 Sunday 124 2012


2868 ENDO12L_M18 MEET-THE-PROFESSOR: CLINICAL - Hereditary Pheochromocytoma [amp] Paraganglioma (8:30 AM - 9:15 AM) Hereditary Pheochromocytoma [amp] Paraganglioma Karel Pacak PREB, NICHD, Bethesda, MD Disclosure Incomplete: KP 2012-06-24T08:30:00 Room 370 2012-06-24T00:00:00 1899-12-30T08:30:00 6154 129 933 M18 M2 Sunday 125 2012


2869 ENDO12L_M19 MEET-THE-PROFESSOR: CLINICAL - NASH/NAFLD: How To Treat [amp] Whom To Treat? (8:30 AM - 9:15 AM) NASH/NAFLD: How To Treat [amp] Whom To Treat? Kenneth Cusi University Texas Health Science Center San Antonio, San Antonio, TX Disclosure Incomplete: KC 2012-06-24T08:30:00 Room 352 DEF 2012-06-24T00:00:00 1899-12-30T08:30:00 6152 130 934 M19 L4 Sunday 126 2012


2870 ENDO12L_M20 MEET-THE-PROFESSOR: CLINICAL - Pituitary Incidentaloma (8:30 AM - 9:15 AM) Pituitary Incidentaloma Mark E Molitch Northwestern University Feinberg Medical School, Chicago, IL Disclosure Incomplete: MEM 2012-06-24T08:30:00 Room 342 ABDE 2012-06-24T00:00:00 1899-12-30T08:30:00 6211 131 935 M20 PIT1 Sunday 127 2012


2871 ENDO12L_M21 MEET-THE-PROFESSOR: CLINICAL - Molecular Markers for the Diagnosis of Thyroid Cancer by Fine Needle Aspiration (8:30 AM - 9:15 AM) Role of Molecular Markers in Cancer Jennifer Anne Sipos Ohio State University, Columbus, OH Session supported by: Genzyme Corporation[br][br]Disclosure Incomplete: JAS 2012-06-24T08:30:00 Room 320 2012-06-24T00:00:00 1899-12-30T08:30:00 6159 132 936 M21 TH2 Sunday 128 2012


2872 ENDO12L_M22 MEET-THE-PROFESSOR: CLINICAL - The Very Insulin-Resistant Diabetes Patient: Work Up [amp] Treatment (8:30 AM - 9:15 AM) The Very Insulin-Resistant Diabetes Patient: Work Up [amp] Treatment Lisa R Tannock University of Kentucky, Lexington, KY Session supported by: Lilly USA, LLC[br][br]Nothing to Disclose: LRT 2012-06-24T08:30:00 Room 372 2012-06-24T00:00:00 1899-12-30T08:30:00 6148 133 937 M22 D10 Sunday 129 2012


2873 ENDO12L_M23 MEET-THE-PROFESSOR: CLINICAL - There[apos]s an App for That: Mobile Health in the Management of Diabetes (8:30 AM - 9:15 AM) There[apos]s an App for That: Mobile Health in the Management of Diabetes [amp] Endocrine Conditions Ronald Tamler Mt Sinai Medical Center, New York, NY Disclosures: RT: Consultant, Sanofi-Aventis. 2012-06-24T08:30:00 Room 332 2012-06-24T00:00:00 1899-12-30T08:30:00 6192 134 938 M23 D5 Sunday 130 2012


2874 ENDO12L_PS2-1 PRESIDENTIAL SYMPOSIUM SESSION: CLINICAL - Leptin Revisited (9:30 AM - 11:00 AM) Endogenous Leptin Manipulation Samuel Dagogo-Jack University of Tennessee - Memphis, Memphis, TN There is marked inter-individual variability in circulating leptin levels, even among persons with similar degree of obesity. We hypothesized that the variability in leptin levels among persons of similar adiposity reflects differences in insulin sensitivity. Using euglycemic clamps, we assessed insulin sensitivity in non-diabetic adults stratified by fasting leptin and body mass index. Our data show an inverse correlation between leptin and insulin sensitivity (r= -0.6), and support the conclusion that obese subjects with fasting leptin [lt] 15 ng/ml are [sim]50% more insulin sensitive than BMI-matched controls with leptin [gt] 15 ng/ml. Work from our lab characterized glucocorticoids (Gc) as leptin secretagogues in humans. The significance of the Gc-leptin axis has been investigated in four separate studies. First, the leptin response to Gc occurs at physiological doses[italic]. [/italic]Second, Gc-induced hyperleptinemia is abolished by fasting[italic].[/italic] Thirdly, inhibition of endogenous steroidogenesis decreases leptin production. Fourth, Gc-induced hyperleptinemia modulates food intake in humans. We have compared basal and Gc-stimulated plasma leptin levels across gender and ethnicity. Fasting leptin was higher in African American (AA) women compared to European American (EA) women (50.6 [plusmn] 3.33 vs. 39.5 [plusmn] 3.51 ng/ml, P=0.001) (+SEM), refelecting higher BMI of AA women. Fasting leptin was similar among AA and EA men. The correlation of fasting leptin and total fat mass did not differ by race (r=0.8, P [lt]0.0001 in AA and EA). The peak Gc-stimulated leptin secretory response (%baseline) was greater in AA than EA(227 [plusmn] 9.6 % vs 205 [plusmn] 6.6). Sensitivity analysis showed 29 hyperresponders (15 AA, 14 EA) with delta leptin [gt]90th percentile and 74 hyporesponders (38 AA, 36 EA) at [lt]25th percentile of dynamic leptin secretion. Predictors of dynamic leptin leptin response include fasting leptin (r= -0.29, P[lt]0.0001), total fat mass (r= -0.26, P[lt]0.0001), BMI (r=0.26, P[lt]0.0001). Massively obese subjects showed impaired dynamic leptin secretion in response to Gc as well as insulin. Using Gc as leptin secretagogue, we identified a leptin secretory defect in diabetic patients. This [ldquo]diabetic dysleptinemia[rdquo], by permitting hunger, could exacerbate obesity and insulin resistance and worsen glycemic control in diabetic patients. Initial evidence suggests that [ldquo]diabetic dysleptinemia[rdquo] may be a complication of rather than a contributor to diabetes; however, the mechanisms remain to be elucidated.[br][br]Sources of Research Support: NIH Grants R01 DK067269, MO1 RR00211 and the American Diabetes Association Clinical Research Award.[br][br]Disclosures: SD-J: Principal Investigator, Astra Zeneca, Novo Nordisk, Boehringer Ingelheim; Ad Hoc Consultant, Eli Lilly & Company; Consultant, Merck & Co. 2012-06-24T09:30:00 Room 320 2012-06-24T00:00:00 1899-12-30T09:30:00 2438 135 939 PS2-1 SSS02 Sunday 131 2012


2875 ENDO12L_PS2-2 PRESIDENTIAL SYMPOSIUM SESSION: CLINICAL - Leptin Revisited (9:30 AM - 11:00 AM) Treatment of Hypothalamic Amenorrhea Corrine Kolka Welt Massachusetts General Hospital, Boston, MA Functional hypothalamic amenorrhea (HA) is caused by stresses that lead to low energy availability, which disrupt hypothalamic GnRH secretion and reproductive function. In addition, rare variants in genes associated with idiopathic hypogonadotropic hypogonadism are found in women with HA and may contribute to the variable susceptibility of women to HA. Leptin administration for the relative leptin deficiency in women with hypothalamic amenorrhea appears to improve reproductive, thyroid, and growth hormone axes and markers of bone formation. Dosing or mode of administration may alleviate some of the weight loss associated with leptin treatment in these women. Standard treatment should always begin with alleviating the stressors that resulted in HA. In addition, estrogen replacement and pulsatile GnRH or gonadotropin therapy can be considered depending on the desire for pregnancy.[br][br]Nothing to Disclose: CKW 2012-06-24T10:00:00 Room 320 2012-06-24T00:00:00 1899-12-30T10:00:00 2476 135 940 PS2-2 SSS02 Sunday 132 2012


2876 ENDO12L_PS2-3 PRESIDENTIAL SYMPOSIUM SESSION: CLINICAL - Leptin Revisited (9:30 AM - 11:00 AM) Treatment in Type 1 Diabetes Roger H Unger University of Texas Southwestern Medical Center, Dallas, TX Here we show that glucagon, not insulin lack, causes all catabolic features of insulin deficiency. First, glucagon suppressors, such as leptin, somatostatin, GLP1 analogs and GABA, suppress the metabolic manifestations of type 1 diabetes. Second, total [beta]-cell destruction does not cause diabetes or impair glucose tolerance in glucagon-receptor null mice unless the receptor is restored by adenoviral delivery, in which severe diabetes appears. Third, spontaneous disappearance of the receptor reverses the diabetes. Fourth, anti-insulin serum causes marked hyperglucagonemia, evidence of paracrine suppression of glucagon by undiluted intra-islet insulin. We conclude 1) that glucose-responsive [beta]-cells normally regulate juxtaposed [alpha]-cells; 2) without intra-islet insulin, unregulated [alpha]-cells hypersecrete glucagon, which directly causes the diabetes; 3) peripherally injected insulin cannot match the high intra-islet levels of insulin without grossly over-insulizing its peripheral targets; 4) therefore, optimal glucoregulation will require physiologic peripheral insulin levels coupled with glucagon suppression by a non-insulin suppressor.[br][br]Nothing to Disclose: RHU 2012-06-24T10:30:00 Room 320 2012-06-24T00:00:00 1899-12-30T10:30:00 2541 135 941 PS2-3 SSS02 Sunday 133 2012


2877 ENDO12L_S19-1 SYMPOSIUM SESSION: TRANSLATIONAL - B-Cell Preservation (T1 [amp] T2) (9:30 AM - 11:00 AM) Basic Physiology of Islet [amp] Beta Cell Function in Diabetes, Emphasizing the Adverse Effects of Chronic Hyperglycemia John (Jack) L Leahy University of Vermont College of Medicine, Colchester, VT Session supported by: Amylin Pharmaceuticals, Inc and Lilly USA, LLC [amp] Novo Nordisk Inc.[br][br]Disclosures: JLL: Advisory Group Member, Aventis Pharmaceuticals, Bristol-Myers Squibb, Jansen Pharmaceuticals, Merck & Co., Novo Nordisk, Takeda. 2012-06-24T09:30:00 Theater C 2012-06-24T00:00:00 1899-12-30T09:30:00 6094 136 942 S19-1 T09-S02 Sunday 135 2012


2878 ENDO12L_S19-2 SYMPOSIUM SESSION: TRANSLATIONAL - B-Cell Preservation (T1 [amp] T2) (9:30 AM - 11:00 AM) Clinical Trials Involving Immune Modulation [amp] Other Mechanisms To Prevent Beta Cell Loss in Type 1 Diabetes Jennifer Byrne Marks University of Miami Miller School, Miami, FL Type 1 diabetes arises from the destruction of [beta]-cells in pancreatic islets, through a selective, immunologically mediated mechanism, resulting in insulin deficiency (1). This occurs in genetically susceptibility individuals and is likely triggered by as yet unidentified environmental factors. The process evolves over years, and is presumably a specific reaction to one or more [beta]-cell proteins or autoantigens. A secondary humoral response is heralded by the appearance in the circulation of autoantibodies that serve as markers of the destructive immune process. Clinical type 1 diabetes becomes evident when the majority of [beta]-cell function is lost and presumably most [beta]-cells are destroyed.[br]Investigation to date has been directed both at interrupting the type 1 diabetes disease process during its stage of evolution and at the time of disease onset.[br]The goal of intervention before overt disease onset is to stop the immune destruction and delay or prevent clinical disease. The ability to identify and predict type 1 diabetes risk using a combination of genetic, immunologic, and metabolic parameters has led to studies designed to explore whether the disease can be prevented by intervening in individuals with autoimmunity. To date, such studies have been without clear success for a number of reasons - therapies targeted at one specific component of the immune system, or one specific autoantigen, may be inadequate to halt the disease process, and a combination-of-interventions approach may be needed.[br]The goal of intervention at or soon after diagnosis of clinical disease is preservation of residual [beta]-cell function, which will presumably result in easier glycemic management and reduce hypoglycemia and chronic complications. A number of interventions has been explored to date (2), some of which have shown potential benefit but were limited by toxicity or short duration of benefit. Others have failed to show efficacy, while others have shown mixed results.[br]If success is to be lasting, it will be necessary to include an approach which involves agents that enhance [beta]-cell function and potentially stimulate repair, regeneration, or neogenesis of [beta]-cells, or a process that protects [beta]-cells from attack.[br]Much work remains before type 1 diabetes is preventable, but the potential to prevent the disease is within reach. Despite current impediments, the progress in recent years has been greater than in the past, and we must be up to the challenge of overcoming those that remain.[br][br]Session supported by: Amylin Pharmaceuticals, Inc and Lilly USA, LLC [amp] Novo Nordisk Inc.[br][br](1) Eisenbarth GS, Diabetes 2010;59:759. (2) Rewers M et al., Diabetes Care 2009;32:1769.[br][br]Disclosures: JBM: Investigator, Bristol-Myers Squibb, Eli Lilly & Company, Novartis Pharmaceuticals. 2012-06-24T10:00:00 Theater C 2012-06-24T00:00:00 1899-12-30T10:00:00 2474 136 943 S19-2 T09-S02 Sunday 136 2012


2879 ENDO12L_S19-3 SYMPOSIUM SESSION: TRANSLATIONAL - B-Cell Preservation (T1 [amp] T2) (9:30 AM - 11:00 AM) Beta Cell Function in Type 2 Diabetes, Including the Effects of Medications Such as TZDs [amp] GLP-1 Analogues Ele Ferrannini Consiglio Nazionale delle Rice, Pisa, Italy Session supported by: Amylin Pharmaceuticals, Inc and Lilly USA, LLC [amp] Novo Nordisk Inc.[br][br]Disclosure Incomplete: EF 2012-06-24T10:30:00 Theater C 2012-06-24T00:00:00 1899-12-30T10:30:00 6096 136 944 S19-3 T09-S02 Sunday 137 2012


2880 ENDO12L_S20-1 SYMPOSIUM SESSION: CLINICAL - [apos]Non-Neoplastic[apos] Pituitary Diseases (9:30 AM - 11:00 AM) Rathke[apos]s Cleft Cysts: An Ongoing Problem? Niki Karavitaki Oxford Center for Diabetes, Endocrinology and Metabolism, Oxford, UK Rathke[apos]s cleft cysts (RCCs) are benign, sellar and/or suprasellar lesions originating from the remnants of Rathke[apos]s pouch. They are the most common incidentally discovered sellar lesions and in routine autopsies they are encountered in 13-33% of normal pituitaries. When symptomatic, the presenting manifestations are related to pressure effects to surrounding structures (mainly headaches, visual disturbance, hypopituitarism).[br]The natural history of symptomatic and asymptomatic RCCs is not clear. In series of non-operated presumed RCCs, the percentage of those not showing progress ranges between 26-94% during follow-up periods up to 9 years. The increase in their size is associated with an imbalance between secretion and absorption of the cyst content, intracystic haemorrhage or infection.[br]Small, asymptomatic RCCs do not require surgical intervention but clinical/imaging surveillance is recommended; its duration, at present, is not clear.[br]In symptomatic ones, surgery is indicated aiming to drain the cyst content and safely remove the capsule. Headaches and major visual field defects improve or resolve in 40-100% and 33-100% of the cases, respectively. Partial hypopituitarism may recover post-surgery (14-50% of the cases), whereas panhypopituitarism seldom resolves. Aggressive surgery with resection of the cyst wall is more commonly associated with new endocrine morbidity.[br]Their recurrence rates after surgery and during variable follow-up periods range between 0-33%. (1) Notably, the lowest relapse rates have been described in reports with relative short mean observation time (less than 3 years), whereas in those with longer follow-up, the relapse rate increases. In a recently published series with a mean follow-up of 48 months, the recurrence rate was 48% at 4 years suggesting that the previously thought optimal course of RCCs may not reflect their true prognosis (2). Most of the relapses are detected within the first 5-6 years suggesting that close follow-up with 12-monthly scans for at least 5 years after surgery is required. Risk factors for recurrence include presence of squamous metaplasia in the cyst wall, cyst size, residual cyst demonstrated on postoperative MRI and the presence of inflammation. The data on the extent of resection/aggressiveness of surgery and risk of relapse are not consistent. The management of recurrent symptomatic lesions includes repeat surgery and the role of irradiation in preventing further relapse has not been clarified.[br][br](1)Trifanescu R et al., Clin Endocrinol 2012;76:151. (2)Trifanescu R et al., Eur J Endocrinol 2011;165:33.[br][br]Nothing to Disclose: NK 2012-06-24T09:30:00 Theater A 2012-06-24T00:00:00 1899-12-30T09:30:00 2423 137 945 S20-1 T01-S03 Sunday 139 2012


2881 ENDO12L_S20-2 SYMPOSIUM SESSION: CLINICAL - [apos]Non-Neoplastic[apos] Pituitary Diseases (9:30 AM - 11:00 AM) Head Trauma [amp] Hypopituitarism Fahrettin Kelestimur Erciyes University, Kayseri, Turkey Head trauma (HT) is a worldwide public health problem. Pituitary dysfunction due to HT was first reported 90 years ago and so far it has been accepted as a rare cause of hypopituitarism. For the last 15 years, a number of cross-sectional and prospective studies have revealed that hypopituitarism is a common complication after HT which may be due to car crashes, falls, child abuse, violence, war accidents and sports injuries including boxing and kickboxing. The mechanisms underlying the pituitary dysfunction after HT have not been fully clarified. Direct mechanical trauma to the hypothalamo-pituitary region, vascular insult, hypoxic damage, autoimmunity and genetic predisposition are among the most commonly proposed explanations for hypothalamo-pituitary injury leading to hormonal abnormalities. Clinico-radiological data suggest that diffuse axonal injury and basal skull fracture are associated with increased prevalence of HT-induced hypopituitarism. The frequency of hypopituitarism in HT patients with positive anti-pituitary antibodies has been found to be significantly higher than in the anti-pituitary-antibodies negative HT patients. Since hypopituitarism is related to the anti-hypothalamic-antibodies instead of anti-pituitary-antibodies in boxers and kickboxers, it seems that hypothalamic damage in sports-related injuries may be more important than pituitary damage. Lower prevalence of hypopituitarism in subjects with APO E3/E3 than the subjects without APO E3/E3 genotype indicates that genetic predisposition plays a role in the development of pituitary dysfunction after HT. Pituitary function may improve over time in a considerable number of patients with HT, but although rarely, may also worsen over the years. The prevalence of hypopituitarism is variable among the studies and ranges from 2% to 35% being GH deficiency the commonest. A great number of HT patients with hypopituitarism are underdiagnosed and therefore untreated. The most suitable time for hormonal investigation is one year after HT and the best candidates for screening are the patients with moderate or severe HT. However, those patients with mild HT who need hospitalization for at least 24 h are also at risk of hypopituitarism and needs screening.[br][br]Nothing to Disclose: FK 2012-06-24T10:00:00 Theater A 2012-06-24T00:00:00 1899-12-30T10:00:00 2437 137 946 S20-2 T01-S03 Sunday 140 2012


2882 ENDO12L_S20-3 SYMPOSIUM SESSION: CLINICAL - [apos]Non-Neoplastic[apos] Pituitary Diseases (9:30 AM - 11:00 AM) Drug-Induced Lymphocytic Hypophysitis [amp] the Hypophysitis Spectrum Maria Fleseriu Oregon Health and Science University, Portland, OR Autoimmune hypophysitis, initially thought to be an autoimmune phenomenon occurring primarily in postpartum women, is now known to be linked to other phenomena affecting both men and women at every age. Autoimmune hypophysitis is classified based on anatomic location, etiology, and histopathological appearance.[br]Clinical manifestations include anterior hypopituitarism with a characteristic pattern of ACTH deficiency, followed by TSH, gonadotrophs and PRL deficiency or hyperprolactinemia. In addition, diabetes insipidus could be an important feature. MRI typically shows symmetrical and homogenous contrast enhancement with a thickened stalk.[br]The current classification includes two new additions to the spectrum of autoimmune hypophysitis: drug- induced autoimmune hypophysitis (Ipilimumab and Ribavirin) and immunoglobulin G4 (IGG- 4) associated autoimmune hypophysitis.[br]Ipilimumab, a cytotoxic T-lymphocyte antigen-4 (CTLA-4) antibody, was recently approved by the FDA for treatment of metastatic melanoma. This therapy represents a novel approach to cancer treatment via disruption of immune tolerance to antigens on tumor cells. The cost of this disruption, however, is a host of immune related adverse events (IRAEs); in clinical trials, autoimmune hypophysitis was reported in up to 18 % of patients. In acute cases, the immediate threat is compromised vision and development of hypopituitarism. A pituitary tumor must be excluded. The initial symptoms may be subtle and clinicians will need a high index of suspicion and an early MRI scan to detect pituitary enlargement before the onset of complete pituitary failure. The current treatment is high-dose glucocorticoid administration and/or other immunosuppressive therapies. The optimal dose and duration of these therapies is not completely defined. There is no clear evidence that glucocorticoid therapy adversely impacts CTLA-4 treatment efficacy, but more studies are needed.[br]A second new clinical entity is IgG4-related autoimmune hypophysitis. IgG4- related plasmacytic disease is characterized by elevated serum IgG4 and tissue infiltration by IgG4-positive plasma cells. This disease is very responsive to glucocorticoid therapy.[br]Drug related autoimmune hypophysitis is a challenging new entity. Familiarity with the full spectrum of IRAEs associated with these novel therapies is important. Continued patient surveillance is essential, as relapses and development of subsequent hormonal deficits occur.[br][br]Nothing to Disclose: MF 2012-06-24T10:30:00 Theater A 2012-06-24T00:00:00 1899-12-30T10:30:00 2406 137 947 S20-3 T01-S03 Sunday 141 2012


2883 ENDO12L_S21-2 SYMPOSIUM SESSION: TRANSLATIONAL - Notch Signaling in Skeletal Metabolism [amp] Pathophysiology (9:30 AM - 11:00 AM) Notch Signaling Mutations in Human [amp] Mouse Skeletal Diseases Jianning Tao Baylor College of Medicine, Houston, TX Notch signaling is an evolutionarily conserved ligand-mediated transmembrane pathway that governs cell fate specification, differentiation, and patterning. Dysregulation of Notch signaling results in a wide range of birth defects, adult onset diseases, and cancers. During the last two decades, the number of components in mammalian Notch signaling pathway has increased to more than fifty. They include Notch receptors, ligands, effectors, bHLH targets, and enzymatic modifiers. Human genetic studies showed that some of those components are associated with disorders in diverse organs such as the skeleton. The natural history of these disorders suggested that Notch signaling may play an important role in postnatal skeletal homeostasis. Indeed, we and others have since shown that Notch signaling is important in osteoblast and osteoclast differentiation in genetically engineered mouse models. Recent reports of Notch mutations in human skeletal dysplasias have corroborated the predictions of mechanistic studies. Moreover, dysregulation of Notch signaling also contributes to the pathogenesis of human and mouse osteosarcoma, the most common primary bone cancer. Together, our and others[apos] studies provide genetic evidence that the effects of Notch signaling are context dependent with different consequences in chondrocytes verse osteoblasts. This may be explained in part by the actions of canonical or Rbpj-dependent and of poorly characterized noncanonical effectors of Notch signaling. Ongoing studies on the relative contribution of noncanonical verse canonical signaling will provide insight on pathogenesis of Notch-related human skeletal diseases.[br][br]Sources of Research Support: Cancer Prevention Institute of Texas Grant RP101017 awarded to BL; NIH Grant AR061565 awarded to JT.[br][br]Nothing to Disclose: JT 2012-06-24T10:00:00 Room 342 ABDE 2012-06-24T00:00:00 1899-12-30T10:00:00 2440 138 948 S21-2 T03-S06 Sunday 144 2012


2884 ENDO12L_S21-3 SYMPOSIUM SESSION: TRANSLATIONAL - Notch Signaling in Skeletal Metabolism [amp] Pathophysiology (9:30 AM - 11:00 AM) Notch Signaling [amp] Skeletal Malignancies Dennis PM Hughes Children[apos]s Cancer Hospital at MD Anderson Cancer Center, Houston, TX Disclosure Incomplete: DPMH 2012-06-24T10:30:00 Room 342 ABDE 2012-06-24T00:00:00 1899-12-30T10:30:00 6040 138 949 S21-3 T03-S06 Sunday 145 2012


2885 ENDO12L_S22-1 SYMPOSIUM SESSION: TRANSLATIONAL - Novel Therapeutic Approaches to Metastatic Thyroid Cancer (English) (9:30 AM - 11:00 AM) Signaling Pathways in Thyroid Cancer: Potential Therapeutic Targets Garcilaso Riesco-Eizaguirre Hospital University Ersitario La Paz, Madrid, Spain Session supported by: Genzyme Corporation[br][br]Disclosure Incomplete: GR-E 2012-06-24T09:30:00 Theater B 2012-06-24T00:00:00 1899-12-30T09:30:00 6015 139 950 S22-1 T02-S02 Sunday 147 2012


2886 ENDO12L_S22-2 SYMPOSIUM SESSION: TRANSLATIONAL - Novel Therapeutic Approaches to Metastatic Thyroid Cancer (English) (9:30 AM - 11:00 AM) Targeting MAPK Signaling: Promises [amp] Challenges James A Fagin Memorial Sloan-Kettering Cancer Center, New York, NY Session supported by: Genzyme Corporation[br][br]Disclosure Incomplete: JAF 2012-06-24T10:00:00 Theater B 2012-06-24T00:00:00 1899-12-30T10:00:00 6016 139 951 S22-2 T02-S02 Sunday 148 2012


2887 ENDO12L_S22-3 SYMPOSIUM SESSION: TRANSLATIONAL - Novel Therapeutic Approaches to Metastatic Thyroid Cancer (English) (9:30 AM - 11:00 AM) New Therapies [amp] Clinical Trials in Medullary [amp] Differentiated Thyroid Cancer Nelson Wohllk Universidad de Chile, Santiago, Chile Options for treatment in differentiated (DTC) or medullary thyroid cancer (MTC) include the use of radioiodine, targeted surgical reduction, radiotherapy and chemotherapy; these last 2 options with little effect. Metastatic DTC concentrates iodine poorly as the disease becomes progressively less differentiated. Patients with progressive MTC or DTC should be enrolled in ongoing clinical trials (CT) of kinase inhibitors (1). In phase III, the primary endpoint is to compare the effect of the drug and placebo on the Progression-Free Survival (PFS) according to RECIST criteria (2). Vandetanib, an inhibitor of RET kinase, VEGFR, and EGFR signaling, has previously shown antitumor activity in a phase II study of patients with advanced hereditary MTC. There is only one phase III trial in patients with progressive MTC that has been completed to date. This trial with vandetanib showed that median PFS was 11 months longer in the group randomly assigned to vandetanib and 45% had an objective rate response (3). Vandetanib was approved by FDA on April 2011. Cabozantinib (XL184), is a potent dual inhibitor of the MET and VEGF pathways designed to block MET driven tumor escape. The ongoing phase III clinical trial of cabozantinib in patients with MTC, known as the EXAM trial met its primary endpoint of improving PFS. Cabozantinib significantly improved median PFS by 7.2 months compared with placebo. In DTC there is one drug ongoing in phase III: E7080(Lenvatinib) in 131I-Refractory DTC(4). A phase II trial studied the effect of lenvatinib (which targets VEGFR1-3, FGFR 1-4, RET, c Kit and PDGFRBeta) in 58 patients with progressive, radioiodine-resistant DTC. Partial response was observed in 50% of patients, and median PFS was 12.6 months (5). Sorafenib (inhibitor of VEGFR2, VEGFR3, RET and BRAF) and sunitinib (inhibitor of all three VEGFRs, RET, and RET/PTC subtypes 1 and 3) have already been approved by FDA and EMEA in other tumors, thus potentially available for off-label use in patients with metastatic DTC and MTC. Both drugs have been mostly associated with stable disease in patients with MTC and DTC (6). Pazopanib is an oral multikinase inhibitor. Phase II studies of pazopanib for MTC are ongoing. There are some other phase II CT, using sorafenib plus everolimus or tenserolimus, everolimus, etc (7). Adverse events for any kinase inhibitors occur frequently, including diarrhea, rash, hypertension, QTc prolongation, dermatitis acneiform/Acne, fatigue and hypothyroidism.[br][br]Session supported by: Genzyme Corporation[br][br]1.-Gild, M. L. et al. Nat. Rev. Endocrinol 2011 Aug 23;7:617-24. 2.-Therasse P. J Natl Cancer Inst 2000; 92:205-16. 3.-Wells SA Jr, et al. J Clin Oncol 2011; 28:767-772. 4.-http://www.exelixis.com/cabozantinib/clinical-trial-information/medullary-thyroid-cancer. 5.-Sherman, S. I. et al. A phase II trial of the multitargeted kinase inhibitor E7080 in advanced radioiodine (RAI)-refractory differentiated thyroid cancer (DTC) [abstract]. J. Clin. Oncol. 29, a5503 (2011). 6.-Cabanillas, M. E. et al. J. Clin. Endocrinol. Metab. 95, 2588-2595 (2010). 7.- http://www.clinicaltrials.gov/.[br][br]Nothing to Disclose: NW 2012-06-24T10:30:00 Theater B 2012-06-24T00:00:00 1899-12-30T10:30:00 2525 139 952 S22-3 T02-S02 Sunday 149 2012


2888 ENDO12L_S23-1 SYMPOSIUM SESSION: TRANSLATIONAL - Nuclear Receptors [amp] Metabolic Disease (9:30 AM - 11:00 AM) Nuclear Receptors Versus Diabetes David D Moore Baylor College of Medicine, Houston, TX Several pathways and pathologies have been suggested as mediators of the connection between obesity and diabetes. These include inflammation in adipose and other tissues, general lipotoxicity and specific toxic lipids, endoplasmic reticulum stress, and steatosis. One specific proposal is that insulin resistance induces a vicious cycle in which hyperinsulinemia increases hepatic lipogenesis and steatosis, which is associated with whole body insulin resistance. The subsequent increase in insulin levels further induces lipogenesis, exacerbates fatty liver and further drives insulin resistance. The remarkable fact that activation of least seven very different nuclear receptors results in antidiabetic effects raises the question of whether there is a common mechanistic thread that connects these effects. Our own results along with those from many other laboratories suggest that the self reinforcing reversal of this vicious cycle via suppression of the lipogenic transcription factor SREBP-1c is such a common thread linking the antidiabetic actions of CAR, LRH-1, TR[beta], ER[alpha] and FXR/SHP. Suppressing hepatic lipogenesis by inhibiting SREBP-1c presents an attractive approach for treating not only steatosis, but also insulin resistance.[br][br]Sources of Research Support: NIH Grants DK46546 and DK085372, and USDA ARS grant 6250-52000-055 awarded to DDM, and the Diabetes and Endocrinology Research Center (P30 DK-079638) at Baylor College of Medicine.[br][br]Nothing to Disclose: DDM 2012-06-24T09:30:00 Room 310 2012-06-24T00:00:00 1899-12-30T09:30:00 172 140 953 S23-1 T08-S04 Sunday 151 2012


2889 ENDO12L_S23-2 SYMPOSIUM SESSION: TRANSLATIONAL - Nuclear Receptors [amp] Metabolic Disease (9:30 AM - 11:00 AM) Glucocorticoids, Cholesterol Sensors [amp] Diabetes Carolyn L Cummins University of Toronto, Toronto, Canada Although widely prescribed for their potent anti-inflammatory actions, glucocorticoid drugs (e.g., dexamethasone) cause undesirable side effects that are features of the metabolic syndrome, including hyperglycemia, fatty liver, insulin resistance, and Type II diabetes. LXRs (liver x receptors) are nuclear receptors that respond to cholesterol metabolites and regulate the expression of a subset of glucocorticoid target genes. We have previously shown that LXR[beta] is required to mediate many of the negative side-effects of glucocorticoids (1). Mice lacking LXR[beta] (but not LXR[alpha]) are resistant to dexamethasone-induced hyperglycemia, hyperinsulinemia, and hepatic steatosis, but remain sensitive to dexamethasone-dependent repression of the immune system. In vivo and in vitro studies found that LXR[beta] promotes these hepatic effects by potentiating dexamethasone-induced expression of the key hepatic gluconeogenic gene, phosphoenolpyruvate carboxykinase. This unexpected relationship between LXR[beta] and the glucocorticoid/GR axis suggested a new avenue for the creation of safer glucocorticoid drugs through a mechanism of selective glucocorticoid receptor transactivation. Using chemical probes targeting LXR[beta], we will test whether partial agonists or antagonists of LXR are able to disrupt the GR/LXR[beta] interplay required for the maximal induction of PEPCK by Dex. Here, we present our unpublished data using a potent pan-LXR antagonist (tested in LXR[alpha]-/- mice) showing that this strategy is effective at inhibiting PEPCK induction by Dex. Furthermore, our data using chromatin IP assays indicate that the mechanism by which LXR[beta] antagonists inhibit GC-induced transactivation may be distinct from the mechanism by which LXR[beta] loss of function results in the same outcome.[br][br](1) Patel R. et al. J Clin Invest 2011; 121:431.[br][br]Sources of Research Support: Canadian Institutes of Health Research (MOP-97904), Natural Sciences and Engineering Research Council of Canada (356873-08), Canadian Foundation for Innovation.[br][br]Nothing to Disclose: CLC 2012-06-24T10:00:00 Room 310 2012-06-24T00:00:00 1899-12-30T10:00:00 2514 140 954 S23-2 T08-S04 Sunday 152 2012


2890 ENDO12L_S23-3 SYMPOSIUM SESSION: TRANSLATIONAL - Nuclear Receptors [amp] Metabolic Disease (9:30 AM - 11:00 AM) Estrogen-Related Receptors (ERRs) in Physiology and Disease Vincent Giguere McGill University, Montreal, Canada Orphan nuclear receptors, in a manner comparable to classic steroid hormone receptors, regulate key developmental and physiological processes. However, the lack of appropriate pharmacological tools has often hindered the identification and study of their biological functions. In this presentation, I will demonstrate that functional and physiological genomics are effective alternatives to discover biological functions associated with orphan nuclear receptors. Indeed, I will document that these approaches have allowed for the unambiguous identification of the estrogen-related receptors (ERRs) [alpha], [beta] and [gamma] (NR3B1, 2 and 3) as global regulators of cellular energy metabolism. We further show that although the three ERR isoforms control analogous gene networks, each isoform performs unique biological functions in a tissue-specific manner in response to a variety of physiological stressors. Finally, I will discuss how the activity of the three ERR isoforms contributes to the development and progression of metabolic diseases as well as to the adaptation of cancer cells to their unique bioenergetic requirement.[br][br]Sources of Research Support: Canadian Institutes for Health Research; Terry Fox Foundation, GenomeQuebec; Canadian Foundation for Innovation.[br][br]Nothing to Disclose: VG 2012-06-24T10:30:00 Room 310 2012-06-24T00:00:00 1899-12-30T10:30:00 2409 140 955 S23-3 T08-S04 Sunday 153 2012


2891 ENDO12L_S24-1 SYMPOSIUM SESSION: CLINICAL - Obesity [amp] Its Complications: Hope for the Future (9:30 AM - 11:00 AM) Bariatric Surgery: Mechanisms of Surgery-Based Therapy for Obesity and Diabetes, More Than Just Rearranging the Plumbing Marzieh Salehi University of Cincinnati, Cincinnati, OH Bariatric surgeries are the most effective treatment for obesity and yet the mechanisms by which these procedures induce weight loss have not been completely understood. Gastric banding, sleeve gastrectomy, and gastric bypass account for more than 95% of bariatric procedures with various proportions in different countries. Weight loss produced by gastric bypass surgery has been consistently greater and more durable than that produced by gastroplasty procedures independent of the degree of gastric restriction. Also, patients with type 2 diabetes have been reported to have near complete diabetes remission within days of having gastric bypass, well before any significant weight loss. The mechanism for this weight-independent diabetes resolution is consistent with a process rapidly engaged by the gastrointestinal alterations of the surgery. There is accumulating evidence supporting changes in the secretion or action of gastrointestinal hormones to mediate this effect, and recent consideration has been given to neurally activated mechanisms involved in glucose homeostasis or energy balance as well as nutrient processing and sensing. Understanding the physiologic mechanisms whereby bariatric surgeries affect weight loss and glucose homeostasis could lead to development of less invasive options for treatment of obesity or type 2 diabetes.[br][br]Sources of Research Support: American Heart Association, 0635181N, American Society for Metabolic and Bariatric Surgery, National Institute of Health, DK083554, and in part by USPHS, UL1 RR026314 from the National Center for Research Resources, NIH.[br][br]Nothing to Disclose: MS 2012-06-24T09:30:00 Room 332 2012-06-24T00:00:00 1899-12-30T09:30:00 2533 141 956 S24-1 T04-S02 Sunday 155 2012


2892 ENDO12L_S24-2 SYMPOSIUM SESSION: CLINICAL - Obesity [amp] Its Complications: Hope for the Future (9:30 AM - 11:00 AM) Pharmacologic Control of Energy Balance with Single Molecule Gut Hormone Combinations Mark W Sleeman Manash University, Victoria, Australia Disclosure Incomplete: MWS 2012-06-24T10:00:00 Room 332 2012-06-24T00:00:00 1899-12-30T10:00:00 6043 141 957 S24-2 T04-S02 Sunday 156 2012


2893 ENDO12L_S24-3 SYMPOSIUM SESSION: CLINICAL - Obesity [amp] Its Complications: Hope for the Future (9:30 AM - 11:00 AM) Novel Therapy for Non-Alcoholic Fatty Liver Disease Kenneth Cusi University Texas Health Science Center San Antonio, San Antonio, TX Nothing to Disclose: KC 2012-06-24T10:30:00 Room 332 2012-06-24T00:00:00 1899-12-30T10:30:00 6044 141 958 S24-3 T04-S02 Sunday 157 2012


2894 ENDO12L_S25-1 SYMPOSIUM SESSION: TRANSLATIONAL - Preservation of Female [amp] Male Gonadal Functions (9:30 AM - 11:00 AM) [italic]In Vitro[/italic] Maturation from Primordial Follicles: Present Experience and Future Endeavors Mary Beth Zelinski Oregon National Primate Research Center, Beaverton, OR Recent developments in techniques to preserve female fertility following cancer treatment have improved the outlook for patients facing premature infertility due to cancer therapies. Ovarian tissue cryopreservation is the only option for preserving fertility in pre-pubertal girls and premenopausal cancer patients requiring immediate treatment. The combination of successful ovarian tissue cryopreservation with [italic]in vitro[/italic] maturation of preantral follicles to produce competent oocytes is emerging as a potential clinical option for fertility preservation, especially for young patients and those for whom ovarian tissue transplantation poses a risk for reintroduction of malignant cells. Nonhuman primate and human secondary follicles can grow to the antral stage in a 3D culture system using an alginate matrix. Survival of macaque follicles in 3D culture depends on animal age, phase of the menstrual cycle at collection and the presence of FSH in the media. Surviving follicles were heterogeneous with regard to their growth potential to reach the antral stage, and were categorized as non-, slow- and fast-growing follicles. These follicle cohorts also differed in their production of steroid hormones and local paracrine factors (AMH, VEGF) which was stage-dependent. Oocytes derived from antral follicles could reinitiate meiotic maturation, fertilize in vitro, and undergo early embryonic cleavage. Macaque secondary follicles isolated from vitrified ovarian tissue, or vitrified individually, survive, grow, form an antrum and produce steroids in 3D culture, indicating functional preservation post-thaw. Human primordial follicles within cortical pieces can grow to the secondary stage in 6 days when cultured in serum-free media without a supporting matrix. Subsequent isolation and further culture of these preantral follicles to the antral stage can occur within an additional 4 days. Production of developmentally competent human oocytes during this short-term, multi-step culture system is under investigation. Future endeavors will optimize the development of both fresh and cryopreserved preantral follicles during 3D and multi-step cultures to consistently produce antral follicles containing mature oocytes capable of yielding live offspring. A robust system for in vitro human follicle development and oocyte maturation will enhance fertility preservation options for young girls and women faced with gametotoxic cancer therapies.[br][br]Jing Xu, Alison Y. Ting, Richard R. Yeoman, Maralee S. Lawson, and Richard L. Stouffer.[br][br]Sources of Research Support: NIH UL1RR024926, R01-HD058293, R01-HD058294, PL1EB008542, U54 HD018185, 8P51OD011092-53.[br][br]Nothing to Disclose: MBZ 2012-06-24T09:30:00 Room 362 2012-06-24T00:00:00 1899-12-30T09:30:00 2482 142 959 S25-1 T05-S03 Sunday 159 2012


2895 ENDO12L_S25-2 SYMPOSIUM SESSION: TRANSLATIONAL - Preservation of Female [amp] Male Gonadal Functions (9:30 AM - 11:00 AM) Cryopreservation [amp] Ovarian Autotransplantation: Current Practice [amp] Possible Risks W Hamish B Wallace Royal Hospital for Sick Children, Edinburgh, UK Disclosure Incomplete: WHBW 2012-06-24T10:00:00 Room 362 2012-06-24T00:00:00 1899-12-30T10:00:00 6107 142 960 S25-2 T05-S03 Sunday 160 2012


2896 ENDO12L_S25-3 SYMPOSIUM SESSION: TRANSLATIONAL - Preservation of Female [amp] Male Gonadal Functions (9:30 AM - 11:00 AM) Gonadal Function Preservation in Male Cancer Patients Stefan Schlatt University of Muenster, M[uuml]nster, Germany Disclosure Incomplete: SS 2012-06-24T10:30:00 Room 362 2012-06-24T00:00:00 1899-12-30T10:30:00 6055 142 961 S25-3 T05-S03 Sunday 161 2012


2897 ENDO12L_S26-1 SYMPOSIUM SESSION: BASIC - Steroid [amp] Circadian Regulation of Reproductive Neuroendocrine Function (9:30 AM - 11:00 AM) SCN/Kisspeptin Interactions in Surge Generation Horacio O de la Iglesia University of Washington, Seattle, WA Disclosure Incomplete: HOdlI 2012-06-24T09:30:00 Room 361 2012-06-24T00:00:00 1899-12-30T09:30:00 6066 143 962 S26-1 T06-S04 Sunday 163 2012


2898 ENDO12L_S26-2 SYMPOSIUM SESSION: BASIC - Steroid [amp] Circadian Regulation of Reproductive Neuroendocrine Function (9:30 AM - 11:00 AM) Steroid Feedback Control of GnRH/GnIH Jeremy Troy Smith Monash University, Melbourne, Australia The neuroendocrine reproductive axis is governed through meticulous communication between the brain, pituitary, and gonads. At the top of this hierarchical regulatory system is the pulsatile release of gonadotropin-releasing hormone (GnRH) from neurons terminating at the median eminence. In turn, gonadal sex steroids act via negative feedback to modulate GnRH secretion. In females, there is an additional switch to acute estrogen-positive feedback leading to the GnRH/LH surge, which is required for ovulation. Because GnRH neurons do not express the requisite steroid hormone receptors to facilitate feedback, these effects must be transferred by other steroid-sensitive neurons. Kisspeptin and gonadotropin inhibitory hormone (GnIH) are recently characterized neuropeptides that have key roles in the regulation of GnRH secretion, the former proving to be a major conduit for transmission of sex steroid feedback. Kisspeptin neurons are located in several discrete brain regions, including the anteroventral periventricular nucleus (AVPV) and arcuate nucleus (ARC) of the hypothalamus and are major stimulators of GnRH secretion. In turn, kisspeptin neurons are robustly regulated by sex steroids and virtually all co-express sex steroid receptors. Significantly, sex steroids can differentially regulate kisspeptin neurons in a region-specific manner, a finding that has illuminated the cellular mechanism of sex steroid positive and negative feedback regulation of GnRH secretion. GnIH, on the other hand, is an inhibitor of GnRH and gonadotropin secretion and is produced in neurons of the dorsomedial nucleus and ventral paraventricular nucleus. Expression of the GnIH gene is reduced during the follicular phase of the estrous cycle and infusion of the peptide inhibits the pulsatile secretion of luteinizing hormone. This suggests that lowering of GnIH tone is permissive for estrogen action to induce the preovulatory LH surge. The precise site of action of GnIH to mediate these effects has not been defined and may do so via modulating the action of GnRH at the level of the pituitary gonadotrope. GnIH neurons project to the external zone of the median eminence and the peptide is secreted into the hypophysial portal blood. Whether this secretion has a role in the steroid feedback control of GnRH action is yet to be determined.[br][br]Sources of Research Support: The National Health and Medical Research Council of Australia project grants 606538 and 1024346 and the Australian Research Council Future Fellowship FT0990986 and project grant DP120100521.[br][br]Nothing to Disclose: JTS 2012-06-24T10:00:00 Room 361 2012-06-24T00:00:00 1899-12-30T10:00:00 2419 143 963 S26-2 T06-S04 Sunday 164 2012


2899 ENDO12L_S26-3 SYMPOSIUM SESSION: BASIC - Steroid [amp] Circadian Regulation of Reproductive Neuroendocrine Function (9:30 AM - 11:00 AM) Circadian Coordination of Gonadotropin-Inhibitory Hormone in Female Reproductive Function Lance J Kriegsfeld University of California, Berkeley, Berkeley, CA Converging lines of evidence implicate a critical role for circadian timing in successful female reproduction across mammalian species, including humans. Women with irregular work or sleep cycles, for example, exhibit reduced fertility and an increased rate of spontaneous abortions, suggesting the importance of circadian functioning in ovulation and pregnancy maintenance. In Syrian hamsters (Mesocricetus auratus), the preovulatory luteinizing hormone (LH) surge occurs precisely every 96 hours when the master brain clock, the suprachiasmatic nucleus (SCN), stimulates the gonadotropin-releasing hormone (GnRH) system in the presence of high estradiol concentrations (positive feedback). Importantly, during the remainder of the estrous cycle, estradiol inhibits LH release via negative feedback. The neural circuitry and neurochemical systems participating in this coordinated switch from estrogen-mediated negative to positive feedback remain unspecified. Our initial investigations suggested a role for rodent RFamide-related peptide-3(RFRP-3), the mammalian homolog of avian gonadotropin-inhibitory hormone (GnIH), in mediating the negative feedback actions of estrogen. Given this potential role of GnIH, we hypothesized that the SCN coordinates the removal of estradiol negative feedback at the time of the LH surge, at least in part, through circadian-controlled disinhibition of GnIH restraint. Our work to date suggests that the SCN serves a dual role, both coordinating gonadotropic axis stimulation with GnIH-mediated negative feedback disinhibition to initiate the LH surge and ovulation and points to a novel, circadian-controlled neural circuit that participates in the transition from estradiol negative to positive feedback.[br][br]Sources of Research Support: NIH Grant HD050470.[br][br]Nothing to Disclose: LJK 2012-06-24T10:30:00 Room 361 2012-06-24T00:00:00 1899-12-30T10:30:00 2427 143 964 S26-3 T06-S04 Sunday 165 2012


2900 ENDO12L_NS3 ENDOCRINE NURSES SYMPOSIA: CLINICAL - Primary Hyperparathyroidism: A Journey over Time (8:30 AM - 9:30 AM) Hyperparathyroidism Camilo Jimenez University of Texas MD Anderson Cancer Center, Houston, TX Disclosure Incomplete: CJ 2012-06-24T09:45:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T09:45:00 6232 124 926 NS3 ENS3 Sunday 166 2012


2901 ENDO12L_NS4 ENDOCRINE NURSES SYMPOSIA: CLINICAL - Emerging Diabetes Therapies (9:45 AM - 10:45 AM) Emerging Diabetes Therapies Saundra Hendricks The Methodist Hospital, Houston, TX Disclosure Incomplete: SH 2012-06-24T10:45:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T10:45:00 6233 144 965 NS4 ENS4 Sunday 167 2012


2902 ENDO12L_Y1 ENDOCRINE YEAR IN BASIC/CLINICAL SCIENCE: CLINICAL - The Year in Diabetes (11:15 AM - 12:00 PM) The Year in Diabetes Robert S Sherwin Yale University School of Medicine, New Haven, CT Disclosures: RSS: Consultant, Amylin Pharmaceuticals, Johnson & Johnson, Mannkind, McKinsey and Company, Novartis Pharmaceuticals, Pfizer, Inc., Bristol-Myers Squibb; Owner, insulet. 2012-06-24T11:15:00 Grand Ballroom AB 2012-06-24T00:00:00 1899-12-30T11:15:00 6135 156 1026 Y1 YI1 Sunday 168 2012


2903 ENDO12L_CDW3-1 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Careers in Industry (11:30 AM - 12:45 PM) Careers in Industry Merav Baz Hecht Bristol-Myers Squibb, Boston, MA Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: MBH 2012-06-24T11:30:00 Trainee Career Center 2012-06-24T00:00:00 1899-12-30T11:30:00 6217 157 1027 CDW3-1 CDW2 Sunday 169 2012


2904 ENDO12L_CDW3-2 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Careers in Industry (11:30 AM - 12:45 PM) Careers in Industry Scott Struthers Crinetics Pharmaceuticals Inc, San Diego, CA Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: SS 2012-06-24T11:30:00 Trainee Career Center 2012-06-24T00:00:00 1899-12-30T11:30:00 6218 157 1028 CDW3-2 CDW2 Sunday 170 2012


2905 ENDO12L_CDW3-3 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Careers in Industry (11:30 AM - 12:45 PM) Careers in Industry David G Maggs Amylin Pharmaceuticals, Inc, San Diego, CA Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: DGM 2012-06-24T11:30:00 Trainee Career Center 2012-06-24T00:00:00 1899-12-30T11:30:00 6219 157 1029 CDW3-3 CDW2 Sunday 171 2012


2906 ENDO12L_CDW3-4 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Careers in Industry (11:30 AM - 12:45 PM) Careers in Industry Frank A Quinn Abbott Diagnostics, Abbott Park, IL Session supported by: Novo Nordisk Inc. [br][br]Disclosure Incomplete: FAQ 2012-06-24T11:30:00 Trainee Career Center 2012-06-24T00:00:00 1899-12-30T11:30:00 6220 157 1030 CDW3-4 CDW2 Sunday 172 2012


2907 ENDO12L_W1-1 WORKSHOP: CLINICAL - The PATH to Accurate Hormone Assays: Testosterone, Estradiol [amp] Vitamin D (12:00 PM - 1:00 PM) Current Status of The Partnership for the Accurate Testing of Hormones: The PATH to Truth in Hormone Assays William Rosner St Luke[apos]s/Roosevelt Hospital Center, New York, NY Nothing to Disclose: WR 2012-06-24T12:00:00 Theater A 2012-06-24T00:00:00 1899-12-30T12:00:00 6197 158 1031 W1-1 W1 Sunday 174 2012


2908 ENDO12L_W1-2 WORKSHOP: CLINICAL - The PATH to Accurate Hormone Assays: Testosterone, Estradiol [amp] Vitamin D (12:00 PM - 1:00 PM) Rational Diagnosis of Androgen Deficiency in Men Based on Population-Based Harmonized Reference Ranges Shalender Bhasin Boston University School of Medicine, Weston, MA Disclosure Incomplete: SB 2012-06-24T12:00:00 Theater A 2012-06-24T00:00:00 1899-12-30T12:00:00 6198 158 1032 W1-2 W1 Sunday 175 2012


2909 ENDO12L_W1-3 WORKSHOP: CLINICAL - The PATH to Accurate Hormone Assays: Testosterone, Estradiol [amp] Vitamin D (12:00 PM - 1:00 PM) Performance of Testosterone [amp] Estradiol Assays in the CDC Hormone Standardization Program Hubert W Vesper Centers for Disease Control and Prevention, Atlanta, GA Disclosure Incomplete: HWV 2012-06-24T12:00:00 Theater A 2012-06-24T00:00:00 1899-12-30T12:00:00 6199 158 1033 W1-3 W1 Sunday 176 2012


2910 ENDO12L_W1-4 WORKSHOP: CLINICAL - The PATH to Accurate Hormone Assays: Testosterone, Estradiol [amp] Vitamin D (12:00 PM - 1:00 PM) Vitamin D Standardization Program (VDSP): A New International Program To Standardize the Measurement of 25(OH)D Christopher T Sempos NIH, Rockville, MD Disclosure Incomplete: CTS 2012-06-24T12:00:00 Theater A 2012-06-24T00:00:00 1899-12-30T12:00:00 6200 158 1034 W1-4 W1 Sunday 177 2012


2912 ENDO12L_CP1 CLINICAL PRACTICE GUIDELINES: CLINICAL - Osteoporosis in Men (12:15 PM - 1:00 PM) Moderator Sundeep Khosla College of Medicine, Rochester, MN Session supported by: Medtronic Diabetes, Merck [amp] Co., Inc. [amp] sanofi-aventis, U.S.[br][br]Disclosure Incomplete: Author to be determined 2012-06-24T12:15:00 Theater B 2012-06-24T00:00:00 1899-12-30T12:15:00 6267 159 1035 CP1 CPG1 Sunday 179 2012


2913 ENDO12L_MC2-1 MASTER CLINICIAN: CLINICAL - Diagnosis [amp] Treatment of Peripheral Precocious Puberty (12:15 PM - 1:15 PM) Diagnosis [amp] Treatment of Peripheral Precocious Puberty Jean-Claude Carel Pediatric Endocrinology and Diabetes, Paris, France Disclosure Incomplete: J-CC 2012-06-24T12:15:00 Grand Ballroom AB 2012-06-24T00:00:00 1899-12-30T12:15:00 6110 160 1036 MC2-1 MC2 Sunday 181 2012


2914 ENDO12L_MC2-2 MASTER CLINICIAN: CLINICAL - Diagnosis [amp] Treatment of Peripheral Precocious Puberty (12:15 PM - 1:15 PM) Diagnosis [amp] Treatment of Peripheral Precocious Puberty Erica A Eugster Indiana University School of Medicine, Indianapolis, IN Disclosures: EAE: Study Investigator, Astra Zeneca. 2012-06-24T12:15:00 Grand Ballroom AB 2012-06-24T00:00:00 1899-12-30T12:15:00 6111 160 1037 MC2-2 MC2 Sunday 182 2012


2915 ENDO12L_SBS1-1 SPECIAL BASIC SCIENCE SESSION: BASIC - GWAS Versus Extreme Phenotype (1:00 PM - 1:45 PM) Genome-Wide Association Studies Related to Diabetes and/or Metabolism Jose C Florez Harvard Medical School/Massachusetts General Hospital, Boston, MA Over the past five years there has been an explosion of genome-wide association studies (GWAS) for phenotypes related to type 2 diabetes and metabolism. These GWAS have occurred on the background of genotyping arrays populated by common single nucleotide polymorphisms (SNPs), deployed in various cohorts that have coalesced to form large international consortia. As a result, we have begun to accumulate lists of genetic loci that influence type 2 diabetes, related quantitative glycemic traits, adiposity, serum lipid levels and blood pressure. Genome-wide association findings have typically illustrated novel pathways, pointed toward fundamental biology, confirmed prior epidemiological observations, drawn attention to the role of [beta]-cell dysfunction in type 2 diabetes, explained a fraction of disease heritability, tempered our expectations with regard to their use in clinical prediction, and provided possible targets for pharmacotherapy and pharmacogenetic clinical trials. On the other hand, the causal variants have only been identified for a handful of these loci, and a substantial proportion of the heritability of these phenotypes remain unexplained. The latter is likely due to insufficient sample sizes to detect small effects, a nearly exclusive focus on populations of European descent, an imperfect capture of uncommon genetic variants, an incomplete ascertainment of alternate (non-SNP) forms of genetic variation, and the lack of exploration of additional genetic models. As the community embraces complementary approaches that include systematic fine-mapping, custom-made replication, denser genotyping arrays, platforms that focus on functional variation, next-generation sequencing techniques, and expansion to non-European populations, the coming years will continue to witness exponential growth in our understanding of the genetic architecture of metabolic phenotypes. Whether these findings will prove useful in disease prediction or therapeutic decision-making must be tested in rigorously designed clinical trials.[br][br]Disclosures: JCF: Ad Hoc Consultant, Novartis Pharmaceuticals, Lilly USA, LLC, Pfizer, Inc. 2012-06-24T13:00:00 Theater C 2012-06-24T00:00:00 1899-12-30T13:00:00 2493 171 1050 SBS1-1 SBS01 Sunday 183 2012


2916 ENDO12L_SBS1-2 SPECIAL BASIC SCIENCE SESSION: BASIC - GWAS Versus Extreme Phenotype (1:00 PM - 1:45 PM) Genetic Studies of the Extreme Phenotype Jonathan C Cohen Uniiversity of Texas SW Medical Center, Dallas, TX Disclosure Incomplete: JCC 2012-06-24T13:00:00 Theater C 2012-06-24T00:00:00 1899-12-30T13:00:00 6133 171 1051 SBS1-2 SBS01 Sunday 184 2012


2917 ENDO12L_NS5-1 ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (1:00 PM - 2:00 PM) Carbohydrate Counting Lynn C Coppolo Bassett Healthcare, Cherry Valley, NY Disclosure Incomplete: LCC 2012-06-24T13:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T13:00:00 6239 172 1052 NS5-1 ENS5 Sunday 185 2012


2918 ENDO12L_NS5-2 ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (1:00 PM - 2:00 PM) Exercise Beth Lucasey TVAS BioMedical, Inc, Lenexa, KS Disclosure Incomplete: BL 2012-06-24T13:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T13:00:00 6234 172 1053 NS5-2 ENS5 Sunday 186 2012


2919 ENDO12L_NS5-3 ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (1:00 PM - 2:00 PM) Endocrine Stimulation Testing Roberta M Hower Lehigh Valley Hospital, Allentown, PA Disclosure Incomplete: RMH 2012-06-24T13:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T13:00:00 6235 172 1054 NS5-3 ENS5 Sunday 187 2012


2920 ENDO12L_NS5-4 ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (1:00 PM - 2:00 PM) ENDO 101/Pituitary Michelle H Gurel Massachusetts General Hospital, Boston, MA Disclosure Incomplete: MHG 2012-06-24T13:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T13:00:00 6236 172 1055 NS5-4 ENS5 Sunday 188 2012


2921 ENDO12L_NS5-5 ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (1:00 PM - 2:00 PM) Research Margaret E Eckert-Norton SUNY Downstate, Brooklyn, NY Disclosure Incomplete: MEE-N 2012-06-24T13:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T13:00:00 6237 172 1056 NS5-5 ENS5 Sunday 189 2012


2922 ENDO12L_NS5-6 ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (1:00 PM - 2:00 PM) Research Patricia S Via McGuire Veterans Affairs Medical Center, Richmond, VA Disclosure Incomplete: PSV 2012-06-24T13:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T13:00:00 6238 172 1057 NS5-6 ENS5 Sunday 190 2012


2923 ENDO12L_NS5-7 ENDOCRINE NURSES SYMPOSIA: CLINICAL - Breakout Sessions (1:00 PM - 2:00 PM) Insulin Management Elaine McLeod Tennessee Valley Healthcare System, Nashville, TN 2012-06-24T13:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T13:00:00 6291 172 1058 NS5-7 ENS5 Sunday 190 2012


2924 ENDO12L_CDW4 CAREER DEVELOPMENT WORKSHOP: TRANSLATIONAL - Opportunities for International Trainees (2:00 PM - 2:45 PM) Opportunities for International Trainees Zhenqi Liu University of Virginia Health System, Charlottesville, VA Disclosure Incomplete: ZL 2012-06-24T14:00:00 Trainee Career Center 2012-06-24T00:00:00 1899-12-30T14:00:00 6254 205 1756 CDW4 CDW10 Sunday 191 2012


2925 ENDO12L_CMF6-1 CASE MANAGEMENT FORUM: CLINICAL - Inpatient Management of Diabetes (2:45 PM - 3:30 PM) Inpatient Management of Diabetes Andrew J Ahmann Oregon Health Sciences University, Portland, OR Disclosure Incomplete: AJA 2012-06-24T14:45:00 Theater A 2012-06-24T00:00:00 1899-12-30T14:45:00 6209 207 1764 CMF6-1 CMF-D1 Sunday 192 2012


2926 ENDO12L_CMF6-2 CASE MANAGEMENT FORUM: CLINICAL - Inpatient Management of Diabetes (2:45 PM - 3:30 PM) Inpatient Management of Diabetes Guillermo Enrique Umpierrez Emory University School of Medicine, Atlanta, GA Nothing to Disclose: GEU 2012-06-24T14:45:00 Theater A 2012-06-24T00:00:00 1899-12-30T14:45:00 6187 207 1765 CMF6-2 CMF-D1 Sunday 193 2012


2927 ENDO12L_CMF7-1 CASE MANAGEMENT FORUM: CLINICAL - Reconciling Diabetes Treatment Algorithms Based on Evidence (2:45 PM - 3:30 PM) Reconciling Diabetes Treatment Algorithms Based on Evidence Lawrence Blonde Ochsner Medical Center, New Orleans, LA Session supported by: Lilly USA, LLC[br][br]Disclosures: LB: Principal Investigator, Eli Lilly & Company Novo Nordisk, Sanofi-Aventis; Speaker, Novo Nordisk, Boehringer Ingelheim, Merck & Co., Johnson & Johnson Diabetes Institute, L.L.C., SANTARUS; Scientific Board Member, Novo Nordisk, GlaxoSmithKline, Merck & Co., Sanofi-Aventis; Consultant, Amylin Pharmaceuticals, SANTARUS; Speaker Bureau Member, Amylin Pharmaceuticals; Owner, Pfizer, Inc., Amylin Pharmaceuticals. 2012-06-24T14:45:00 Room 332 2012-06-24T00:00:00 1899-12-30T14:45:00 6189 208 1766 CMF7-1 CMF-D2 Sunday 194 2012


2928 ENDO12L_CMF7-2 CASE MANAGEMENT FORUM: CLINICAL - Reconciling Diabetes Treatment Algorithms Based on Evidence (2:45 PM - 3:30 PM) Reconciling Diabetes Treatment Algorithms Based on Evidence Jaime A Davidson University of Texas Southwestern Medical Center, Dallas, TX Session supported by: Lilly USA, LLC[br][br]Disclosure Incomplete: JAD 2012-06-24T14:45:00 Room 332 2012-06-24T00:00:00 1899-12-30T14:45:00 6247 208 1767 CMF7-2 CMF-D2 Sunday 195 2012


2929 ENDO12L_M24 MEET-THE-PROFESSOR: CLINICAL - Adrenal Insufficiency (2:45 PM - 3:30 PM) Adrenal Insufficiency Baha M Arafah Case Western Reserve University, Cleveland, OH Nothing to Disclose: BMA 2012-06-24T14:45:00 Room 372 2012-06-24T00:00:00 1899-12-30T14:45:00 6139 209 1768 M24 A5 Sunday 196 2012


2930 ENDO12L_M25 MEET-THE-PROFESSOR: CLINICAL - Changing Sex Hormones in Transgender Persons (2:45 PM - 3:30 PM) Changing Sex Hormones in Transgender Persons Wylie C Hembree College of Physicians and Surgeons, Woodcliff Lake, NJ Nothing to Disclose: WCH 2012-06-24T14:45:00 Room 360 2012-06-24T00:00:00 1899-12-30T14:45:00 6170 210 1769 M25 M4 Sunday 197 2012


2931 ENDO12L_M26 MEET-THE-PROFESSOR: CLINICAL - Glucocorticoid-Induced Osteoporosis (GIOP) (2:45 PM - 3:30 PM) Glucocorticoid-Induced Osteoporosis (GIOP) Robert A Adler McGuire Veterans Affairs Medical Center, Richmond, VA Disclosures: RAA: Investigator, Eli Lilly & Company, Merck & Co., Amgen, Novartis Pharmaceuticals; Principal Investigator, Genentech, Inc. 2012-06-24T14:45:00 Room 351 2012-06-24T00:00:00 1899-12-30T14:45:00 6142 211 1770 M26 B3 Sunday 198 2012


2932 ENDO12L_M27 MEET-THE-PROFESSOR: CLINICAL - Growth Hormone Deficiency (2:45 PM - 3:30 PM) Growth Hormone Deficiency Ken Ho Princess Alexandra Hospital, Brisbane, Australia Nothing to Disclose: KH 2012-06-24T14:45:00 Room 370 2012-06-24T00:00:00 1899-12-30T14:45:00 6157 212 1771 M27 PIT2 Sunday 199 2012


2933 ENDO12L_M28 MEET-THE-PROFESSOR: CLINICAL - Klinefelter Syndrome (2:45 PM - 3:30 PM) Klinefelter Syndrome Ronald S Swerdloff LABioMedicine at Harbor UCLA Medical Center, Torrance, CA Nothing to Disclose: RSS 2012-06-24T14:45:00 Room 371 2012-06-24T00:00:00 1899-12-30T14:45:00 6223 213 1772 M28 MR1 Sunday 200 2012


2934 ENDO12L_M29 MEET-THE-PROFESSOR: CLINICAL - Lp(a) [amp] Other CVD Risks: Worth Checking [amp] in Whom? (2:45 PM - 3:30 PM) Lp(a) [amp] Other CVD Risks: Worth Checking [amp] in Whom? Janet B McGill Washington University in St Louis, Saint Louis, MO Nothing to Disclose: JBM 2012-06-24T14:45:00 Room 310 2012-06-24T00:00:00 1899-12-30T14:45:00 6190 214 1773 M29 L3 Sunday 201 2012


2935 ENDO12L_M30 MEET-THE-PROFESSOR: CLINICAL - Medullary Thyroid Cancer (2:45 PM - 3:30 PM) Medullary Thyroid Cancer Richard T Kloos Ohio State University, Columbus, OH Disclosure Incomplete: RTK 2012-06-24T14:45:00 Room 342 ABDE 2012-06-24T00:00:00 1899-12-30T14:45:00 6162 215 1774 M30 TH5 Sunday 202 2012


2936 ENDO12L_M31 MEET-THE-PROFESSOR: CLINICAL - PCOS: Use of Metformin To Allow Pregnancy: Other Treatments (2:45 PM - 3:30 PM) PCOS: Use of Metformin To Allow Pregnancy: Other Treatments Richard S Legro Pennsylvania State University College of Medicine, Hershey, PA Nothing to Disclose: RSL 2012-06-24T14:45:00 Room 352 DEF 2012-06-24T00:00:00 1899-12-30T14:45:00 6149 216 1775 M31 FR1 Sunday 203 2012


2937 ENDO12L_M4 MEET-THE-PROFESSOR: CLINICAL - Pediatric Bone Disease (2:45 PM - 3:30 PM) Pediatric Bone Disease Erik Allen Imel Indiana University School of Medicine, Indianapolis, IN Nothing to Disclose: EAI 2012-06-24T14:45:00 Room 362 2012-06-24T00:00:00 1899-12-30T14:45:00 6246 219 1780 M4 B8 Sunday 204 2012


2938 ENDO12L_M32 MEET-THE-PROFESSOR: CLINICAL - Pediatric Calcium Disorders (2:45 PM - 3:30 PM) Pediatric Calcium Disorders Rachel Ilana Gafni NIH, Bethesda, MD Nothing to Disclose: RIG 2012-06-24T14:45:00 Room 320 2012-06-24T00:00:00 1899-12-30T14:45:00 6214 217 1776 M32 PED6 Sunday 205 2012


2939 ENDO12L_PMW1-1 PRACTICE MANAGEMENT WORKSHOP: CLINICAL - Managing the Transition of Care for Patients with Type 1 Diabetes (2:45 PM - 3:30 PM) Managing the Transition of Care for Patients with Type 1 Diabetes Mary Carol Greenlee Western Slope Endo, Grand Junction, CO Nothing to Disclose: MCG 2012-06-24T14:45:00 Room 361 2012-06-24T00:00:00 1899-12-30T14:45:00 6206 218 1777 PMW1-1 PMW1 Sunday 206 2012


2940 ENDO12L_PMW1-2 PRACTICE MANAGEMENT WORKSHOP: CLINICAL - Managing the Transition of Care for Patients with Type 1 Diabetes (2:45 PM - 3:30 PM) Managing the Transition of Care for Patients with Type 1 Diabetes Celeste Brickler Hart North Florida Thyroid Center, Tallahassee, FL Disclosure Incomplete: CBH 2012-06-24T14:45:00 Room 361 2012-06-24T00:00:00 1899-12-30T14:45:00 6212 218 1778 PMW1-2 PMW1 Sunday 207 2012


2941 ENDO12L_PMW1-3 PRACTICE MANAGEMENT WORKSHOP: CLINICAL - Managing the Transition of Care for Patients with Type 1 Diabetes (2:45 PM - 3:30 PM) Managing the Transition of Care for Patients with Type 1 Diabetes Katharine Creskoff Garvey Children[apos]s Hospital Boston, Boston, MA Disclosure Incomplete: KCG 2012-06-24T14:45:00 Room 361 2012-06-24T00:00:00 1899-12-30T14:45:00 6213 218 1779 PMW1-3 PMW1 Sunday 208 2012


2942 ENDO12L_NS6 ENDOCRINE NURSES SYMPOSIA: CLINICAL - Body Composition Changes [amp] Use of GHRH in HIV Lipodystrophy (10:45 AM - 11:45 AM) GHRH/HIV Lypodystrophy Steven Kyle Grinspoon Massachusetts General Hospital, Boston, MA Disclosure Incomplete: SKG 2012-06-24T15:15:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T15:15:00 6240 145 966 NS6 ENS6 Sunday 209 2012


2943 ENDO12L_PS3-1 PRESIDENTIAL SYMPOSIUM SESSION: CLINICAL - Geographic [amp] Ethnic Variations in Fracture Risk: Clinical Implications (3:45 PM - 5:15 PM) Variation in Fracture Rates: Impact on FRAX Marjorie Luckey St Barnabas Osteoporosis and Metabolic Bone Diseases Center, Livingstone, NJ Disclosure Incomplete: ML 2012-06-24T15:45:00 Grand Ballroom AB 2012-06-24T00:00:00 1899-12-30T15:45:00 6032 221 1782 PS3-1 T03-S01 Sunday 211 2012


2944 ENDO12L_PS3-2 PRESIDENTIAL SYMPOSIUM SESSION: CLINICAL - Geographic [amp] Ethnic Variations in Fracture Risk: Clinical Implications (3:45 PM - 5:15 PM) BMD Variation across Ethnicities [amp] Populations William D Leslie St Boniface General Hospital, Winnipeg, Canada Differences in bone mineral density (BMD) have been observed between ethnic groups, but these only partially account for observed variations in fracture rates. The basis for these variations is complex, and contributing factors include ethnic differences in body size/composition, skeletal size, genetics, lifestyle, and disease comorbidities. Despite this, the gradient of risk for fracture from BMD and other clinical risk factors appears to be similar across ethnic groups. This presentation will explore factors associated with ethnicity and how these may alter the relationship between BMD and fracture risk. Clinical implications for fracture risk assessment are explored in specific populations.[br][br]Nothing to Disclose: WDL 2012-06-24T16:15:00 Grand Ballroom AB 2012-06-24T00:00:00 1899-12-30T16:15:00 2496 221 1783 PS3-2 T03-S01 Sunday 212 2012


2945 ENDO12L_PS3-3 PRESIDENTIAL SYMPOSIUM SESSION: CLINICAL - Geographic [amp] Ethnic Variations in Fracture Risk: Clinical Implications (3:45 PM - 5:15 PM) Insights from New Imaging Techniques Marcella Donovan Walker Columbia University College of Physicians [amp] Surgeons, New York, NY Racial differences in bone mineral density (BMD), as measured by dual x-ray absorptiometry (DXA), do not completely explain racial differences in fracture risk. Differences in other aspects of bone quality may partially account for such discrepancies. Measurement of other facets of bone quality, which independently contribute to bone strength, in addition to BMD holds promise to improve fracture risk prediction in the future. DXA-based techniques can be used to estimate bone geometry and strength. Newer non-invasive imaging technologies, including central and peripheral quantitative computed tomography (QCT), high resolution peripheral quantitative computed tomography (HRpQCT) and micro magnetic resonance imaging ([micro]MRI) can provide information on bone size, geometry, volumetric bone density and bone microarchitecture. Bone structural information obtained using these techniques can be incorporated into finite element models (FEM) which provide estimates of bone strength. Recent work suggests that these techniques are able to discriminate those with and without fracture and elucidate effects of disease states on bone microarchitecture. This presentation will provide an overview of new imaging technologies and review recent investigations which have identified key racial differences in bone size, geometry, volumetric density, microarchitecture and estimated bone strength that may contribute to racial differences in fracture risk.[br][br]Sources of Research Support: NIH Grant K23AR053507, National Osteoporosis Foundation, New York Academy of Medicine [ndash] Mary and David Hoar Grant.[br][br]Nothing to Disclose: MDW 2012-06-24T16:45:00 Grand Ballroom AB 2012-06-24T00:00:00 1899-12-30T16:45:00 2428 221 1784 PS3-3 T03-S01 Sunday 213 2012


2946 ENDO12L_PS4-1 PRESIDENTIAL SYMPOSIUM SESSION: BASIC - Nuclear Receptors in Inflammation [amp] Immune Regulation (3:45 PM - 5:15 PM) The Glucocorticoid Receptor Goes Redneck John A Cidlowski NIEHS/NIH, Research Triangle Pk, NC Glucocorticoids are necessary for life after birth and regulate numerous biological processes in man, including glucose homeostasis, protein catabolism, respiratory function, behavior and inflammation. Medically, synthetic glucocorticoids are widely prescribed as anti-inflammatory agents in the treatment of numerous diseases. The mechanisms mediating these anti-inflammatory actions of glucocorticoids are complex and often tissue- and/or cell-type specific. In the studies presented in this lecture, we describe a newly identified cytoplasmic glucocorticoid receptor-interacting protein, HIC, that modulates the ability of the glucocorticoid receptor to regulate inflammatory genes. Mechanistic studies suggest that HIC has a critical role for glucocorticoid phosphorylation in the process.[br][br]The author would like to acknowledge the following co-authors: John M. Busillo and Robert H. Oakley,Laboratory of Signal Transduction, NIEHS/NIH, Research Triangle Park, North Carolina, USA.[br][br]Nothing to Disclose: JAC 2012-06-24T15:45:00 Room 332 2012-06-24T00:00:00 1899-12-30T15:45:00 2446 222 1785 PS4-1 SSS03 Sunday 215 2012


2947 ENDO12L_PS4-2 PRESIDENTIAL SYMPOSIUM SESSION: BASIC - Nuclear Receptors in Inflammation [amp] Immune Regulation (3:45 PM - 5:15 PM) Glucocorticoid Regulation of Cytokine Gene Expression: The Role of Transcriptional Cofactor GRIP1 Inez Rogatsky Hospital for Special Surgery, New York, NY For decades, the management of inflammatory and autoimmune diseases relied on glucocorticoids (GCs), which repress the transcription of numerous inflammatory mediators, yet the underlying molecular mechanisms remain obscure. In addition, we found that panels of inflammatory genes are activated at distinct stages of the transcription cycle (Adelman et al, PNAS 2009); whether their susceptibility to and/or mechanisms of repression differ accordingly is unknown.[br]We report that in primary mouse macrophages genes encoding inflammatory cytokines were repressed by the liganded glucocorticoid receptor (GR) irrespective of the TLR pathway responsible for the induction, or whether they were activated via RNA Polymerase (Pol)2 recruitment and transcription initiation vs. Pol2 release from the early elongation block. In both cases, GR occupied NFkB binding sites and repressed transcription without affecting p65 occupancy; the transcriptional coregulator GRIP1, suggested to play a role in GR-mediated repression by our earlier studies, was recruited concomitantly with GR. We generated mice enabling a hematopoietic cell-restricted GRIP1 deletion and found that genes of both classes were dose-dependently derepressed as a function of GRIP1 KD. Furthermore, RNA-Seq-based transcriptome analysis revealed an unexpectedly broad derepression of LPS-induced GC-sensitive inflammatory mediators in GRIP1 KD macrophages while GR-activated anti-inflammatory genes were unaffected suggesting that GRIP1 co-repressor activity may be dominant in this context. Corroborating gene expression data, GRIP1 KD mice were sensitized to a systemic LPS challenge implicating GRIP1 in anti-inflammatory actions of GCs in vivo.[br]These previously unrecognized molecular functions of GRIP1 pose a question as to what enables its activating or repressing properties, or selective engagement in physiologically antagonistic pathways. We found that GRIP1 undergoes GC-induced, GR interaction-dependent phosphorylation, identified one constitutive and six inducible phosphorylation sites, and two putative GRIP1 kinases. GRIP1 phosphorylation was conserved across cell types and species, and affected GR-regulated transcription and GRE-specific phospho-GRIP1 recruitment to native GR targets. We propose that cofactor engagement by GR is neither passive nor stochastic; rather, GR actively imparts modifications that dictate GRIP1 function in a subset of complexes adding a layer of specificity to GR transcriptional control.[br][br]Nothing to Disclose: IR 2012-06-24T16:15:00 Room 332 2012-06-24T00:00:00 1899-12-30T16:15:00 2488 222 1786 PS4-2 SSS03 Sunday 216 2012


2948 ENDO12L_PS4-3 PRESIDENTIAL SYMPOSIUM SESSION: BASIC - Nuclear Receptors in Inflammation [amp] Immune Regulation (3:45 PM - 5:15 PM) Nuclear Receptor Signaling in Dendritic Cells and Macrophages from a Genome-Wide Perspective Laszlo Nagy University of Debrecen Medical [amp] Health Science, Debrecen, Hungary A key issue in the immune system is to generate specific cell types often with opposing activities. The mechanisms of differentiation and subtype specification of macrophages and dendritic cells are critical in order to understand the regulatory principles and logic of the immune system. Besides cytokines and pathogens it is increasingly appreciated that lipid signaling also has a key role in differentiation and subtype specification. In recent years we have explored how intracellular lipid signaling via nuclear hormone receptors, regulates cellular differentiation, subtype specification, immune as well as metabolic homeostasis. Based on genome-wide transcriptome analyses it appears that RXR heterodimeric nuclear receptors provide these cells with a coordinated and interrelated network of transcriptional regulators for interpreting the lipid milieu and the metabolic changes to bring about gene expression changes leading to subtype and functional specification. It has been also established that nuclear hormone receptor mediated pathways interact with cytokine signaling to initiate gene expression changes. For example Interleukin 4 mediated STAT6 signaling collaborates with PPARg to regulate alternatively activated macrophage specific transcription. An important question is to explore how these pathways interact at a genome-wide, cistromic level. We have addressed this by determining the genomic localization and cistromic interactions of these transcription factors along with histone modifications and regulated primary transcripts using the combination of ChIP[ndash]Seq, RNA-Seq and GRO-Seq in mouse primary macrophages. The combined analyses of these data along with the emerging model of cistromic interactions will be presented and discussed.[br][br]Nothing to Disclose: LN 2012-06-24T16:45:00 Room 332 2012-06-24T00:00:00 1899-12-30T16:45:00 2477 222 1787 PS4-3 SSS03 Sunday 217 2012


2949 ENDO12L_S27-1 SYMPOSIUM SESSION: BASIC - Getting That GPCR to the Cell Surface (3:45 PM - 5:15 PM) Pharmacoperones: A New Class of Therapeutics P Michael Conn Oregon Health Sciences University, Beaverton, OR G protein-coupled receptors (GPCRs) are plasma membrane proteins that are utilized by cells to mediate responses to sensory stimuli, hormones and neurotransmitters. Together, these receptors constitute the largest family of validated drug targets with over 30% of all drugs targeting them. Mutations in GPCRs are the underlying cause of more than 30 diseases. Mutations lead to misfolded receptors that are recognized by the cell[apos]s quality control system (QCS) and retained in the endoplasmic reticulum, rather than trafficked to the plasma membrane. The assessment by the QCS is based on general chemical criteria (e.g., exposure of hydrophobic plates, unpaired cysteines), rather than functional criteria (e.g. ability to bind ligand, couple to effector). Using a model GPCR, the gonadotropin releasing hormone receptor (GnRHR), our studies show that misfolded GPCR mutants typically retain biological activity and, in most cases, can be restored to function when trafficked to the plasma membrane. This corrects an existing view that loss of function due to mutation results from loss of intrinsic function at the molecular level. The presentation will describe the underlying concepts that led to the development of a new class of drugs [ldquo]pharmacoperones[rdquo] (from pharmacological chaperones) that specifically rescue misrouted proteins, and restore them to function. This work has led to identification of molecular and cellular mechanism of action of these drugs, a high-throughput (HTS) screen for chemical libraries and an animal model which demonstrates the in vivo efficacy of this approach. Because proteins are processed by common mechanisms, this approach applies to receptors, ion channels, enzymes, and other proteins, suggesting that rescue by pharmacoperones might apply to an array of human diseases that result from misfolding, among these: cystic fibrosis, hypogonadotropic hypogonadism, nephrogenic diabetes insipidus, hypercholesterolemia, cataracts, neurodegenerative diseases (Huntington[apos]s, Alzheimer[apos]s, and Parkinson[apos]s diseases) and cancer. In the case of particular proteins, success with a striking number of different mutants supports the view that pharmacoperones will become powerful ammunition in our therapeutic arsenal.[br][br]Sources of Research Support: This work was supported by National Institutes of Health Grants RR030229, DK85040, RR000163 and HD19899. Others who have contributed to this work are: Jo Ann Janovick, M. David Stewart, Richard R. Behringer, Darla L. Jacob and LD Martin.[br][br]Nothing to Disclose: PMC 2012-06-24T15:45:00 Room 342 ABDE 2012-06-24T00:00:00 1899-12-30T15:45:00 2394 223 1788 S27-1 T08-S08 Sunday 219 2012


2950 ENDO12L_S27-2 SYMPOSIUM SESSION: BASIC - Getting That GPCR to the Cell Surface (3:45 PM - 5:15 PM) Role of Endocytosis [amp] Recycling in CRFR-Mediated Sensitization of 5HT2R Signaling Stephen SG Ferguson Robarts Research Institute, London, Canada Depression is a widespread and serious disorder that affects the daily lives of 10-15% of Canadians. There are multiple potential causes of depression, including genetic predisposition, but the majority of cases are influenced by environmental factors. Anatomical localization studies have shown that both serotonin (5HT) 2A receptor (5HT2AR) and corticotropin-releasing factor receptor 1 (CRFR1) are appropriately localized within the brain to regulate each others function at the molecular level. Moreover, several recent studies have reported effects of CRF peptide on 5HT mediated function and signal transduction. However, there is still a lack of information regarding how CRF modulates the 5-HT system at the molecular level. The current presentation will overview the role of GPCR endocytosis, and recycling in the regulation of 5HT2AR signalling following the prior activation of CRFR1. We find that CRFR1 activation leads to the co-internalization of both 5-HT2AR and 5-HT2CR, whereas 5-HT2AR activation does not stimulate CRFR1 endocytosis in either HEK 293 cells or primary mouse (E15-E16) cortical (10 DIV) neurons. We also show that the activation of the CRFR results in a 40 and 47% augmentation in the EMAX for 5-HT-stimulated IP formation in human embryonic kidney (HEK 293) cells co-expressing either human 5-HT2AR or human 5-HT2CR, respectively. This CRFR1-mediated increase in 5-HT receptor signalling is also observed following the activation of endogenous CRF receptors in primary mouse cortical neurons. The augmentation of 5-HT receptor signalling is dependent upon endocytosis as it is not observed in cells expression a dominant-negative dynamin mutant. In addition, CRFR1-mediated increases in 5-HT receptor signalling were blocked by the overexpression of a dominant-negative Rab4 mutant to block receptor recycling. Injection of CRF peptide into the prefrontal cortex of mice 30 minutes prior to challenge with the 5-HT2 receptor agonist DOI also results in enhance anxiety-related behaviours when compared to mice treated with either CRF or DOI alone. Thus, we conclude that the observed CRFR1-mediated increase in 5-HT2AR signalling requires both the endocytosis and recycling of the 5HT2AR and represents a novel mechanism for the regulation of GPCR signal transduction that may be involved in stress induced anxiety-related behaviour responses.[br][br]Nothing to Disclose: SSGF 2012-06-24T16:15:00 Room 342 ABDE 2012-06-24T00:00:00 1899-12-30T16:15:00 2410 223 1789 S27-2 T08-S08 Sunday 220 2012


2951 ENDO12L_S27-3 SYMPOSIUM SESSION: BASIC - Getting That GPCR to the Cell Surface (3:45 PM - 5:15 PM) From Phenotype to Pharmacology: RAMP Mice as GPCR Discovery Tools Kathleen M Caron University of North Carolina-Chapel Hill, Chapel Hill, NC Receptor activity modifying proteins (RAMPs) control G protein-coupled receptor (GPCR) forward-trafficking and recycling, ligand binding affinity and specificity and second messenger signaling. Our lab has taken a genetic and phenotype-driven approach to identify GPCR-RAMP interactions that could serve as pharmacological targets for the treatment of disease conditions. We have primarily focused on the CLR/RAMP interaction with respect to adrenomedullin (AM) signaling; but our laboratory has also developed sophisticated genetic mouse models for all mammalian RAMPs and numerous RAMP-associated GPCRs and their cognate ligands. Using this approach we have identified CLR/RAMP2/AM as the first GPCR signaling paradigm required for normal lymphatic vascular development during embryogenesis. Using inducible Cre-LoxP strategies, we have also recently shown that CLR is required for normal lymphatic function in adult animals. While we consistently find that the CLR/RAMP2/AM signaling axis plays an essential role in conferring cardioprotection and in maintaining normal female reproduction, an expanded repertoire of phenotypes in RAMP2 mice supports the concept that RAMPs have physiological functions that extend well-beyond the canonical CLR receptor. Our work thus far in animal models has helped to define the biological functions of RAMPs under normal and pathological conditions and has paved the way for potential exploitation of the GPCR-RAMP interaction as a unique and pharmacologically tractable interface for novel drug design targeted toward several common and severe cardiovascular disorders.[br][br]Sources of Research Support: NIH grants HL091973, HD060860 and ARRA-HD060860-01S1 (K.M.C.), the March of Dimes Birth Defects Foundation and the Burroughs Wellcome Fund to KMC.[br][br]Nothing to Disclose: KMC 2012-06-24T16:45:00 Room 342 ABDE 2012-06-24T00:00:00 1899-12-30T16:45:00 194 223 1790 S27-3 T08-S08 Sunday 221 2012


2952 ENDO12L_S28-1 SYMPOSIUM SESSION: TRANSLATIONAL - Long Term Effects of GH/IGF-I Reduction (3:45 PM - 5:15 PM) Clues to a Cancer [amp] Diabetes-Free Life Priya Balasubramanian University of Southern California, Los Angeles, CA Session supported by: Ipsen Biopharmaceuticals, Inc.[br][br]Nothing to Disclose: PB 2012-06-24T15:45:00 Room 320 2012-06-24T00:00:00 1899-12-30T15:45:00 6072 224 1791 S28-1 T07-S01 Sunday 223 2012


2953 ENDO12L_S28-2 SYMPOSIUM SESSION: TRANSLATIONAL - Long Term Effects of GH/IGF-I Reduction (3:45 PM - 5:15 PM) Animal Models of the GH/IGF Axis: What Do They Tell Us about Aging? Darlene E Berryman Ohio University, Athens, OH To study aging in mammals, mice are common because of their genetic similarity to humans, their relatively short lifespan, as well as the ability to easily manipulate their genome. Mice with alterations in the GH/IGF axis reveal the importance of this axis in aging. Overall, mice with a reduction in GH action (e.g. Ames, Snell, and growth hormone receptor gene disrupted mice (GHR-/-)) or IGF-1 (e.g. midi-mice, mice heterozygous for deletion of the IGF receptor) have been shown to have extended lifespans and exhibit symptoms of delayed aging as compared to control littermates. In contrast, mice with elevated GH signaling, such as GH transgenic (bGH) mice, have reduced lifespans. Interestingly, there are exceptions to this general trend. For example, growth hormone receptor antagonist transgenic (GHA) mice have a reduction in the action of the GH/IGF axis yet have no alteration in longevity. Comparing and contrasting the resultant physiological phenotypes of these animal models have provided valuable clues to the roles of GH and IGF-1 in aging. Discussed will be a comparison of these mice and the current understanding of how particular traits may influence the progression of aging. Special focus will be placed on the unique phenotype of GHA mice and why they may not experience the benefits in longevity despite a reduction in the GH/IGF-1 axis.[br][br]Session supported by: Ipsen Biopharmaceuticals, Inc.[br][br]The author would like to acknowledge the following co-authors: John J. Kopchick, Edward O. List, Ellen R. Lubbers, Vishakha Magon.[br][br]Sources of Research Support: This work was supported in part by the State of Ohio[apos]s Eminent Scholar Program that includes a gift from Milton and Lawrence Goll, by the AMVETS, by the Diabetes Research Initiative at Ohio University, and by NIH grants DK083729, AG031736.[br][br]Nothing to Disclose: DEB 2012-06-24T16:15:00 Room 320 2012-06-24T00:00:00 1899-12-30T16:15:00 2456 224 1792 S28-2 T07-S01 Sunday 224 2012


2954 ENDO12L_S28-3 SYMPOSIUM SESSION: TRANSLATIONAL - Long Term Effects of GH/IGF-I Reduction (3:45 PM - 5:15 PM) Effects of GHRH Antagonists on Aging in Mice William A Banks Puget Sound Health Care System, Seattle, WA Growth hormone releasing hormone (GHRH), first isolated from a pancreatic tumor, regulates GH and IGF-I release. However, its receptors are widespread throughout the body with direct actions that are independent of GH and IGF-I. GHRH receptors are located in the brain (e.g., hypothalamus, brain stem, cortex), loop of Henle in the kidney, heart myocytes, and reproductive tract. Many cancers release GHRH to act at their own GHRH receptors, stimulating their own growth in autocrine fashion. As such, GHRH receptor antagonists have potential both as pharmacologic probes to better understand GHRH physiology and as therapeutics. GHRH receptor antagonists can cross the blood-brain barrier (BBB), explaining how they can inhibit growth of brain tumors and exert actions on cognition, anxiety, depression, and sleep. We have examined the effects of the potent GHRH antagonist MZ-5-156 on various parameters in the aged SAMP8 mouse, a mouse used as a model of Alzheimer[apos]s disease. The SAMP8 is a natural mutation with a median life span of about 16-18 mo that with aging develops increases in amyloid beta peptide levels and oxidative stress, cholinergic deficits and cognitive impairments, all reversed with antisense directed against amyloid precursor protein. Starting at age 10 mo, SAMP8 mice received daily intraperitoneal injections of either 10 microg/mouse of MZ-5-156 or vehicle. An increase in brain GSH with a reduction in the oxidative enzyme GPx suggested positive effects on brain oxidation. Median life expectancy from start of treatment was increased from 26 weeks to 34 weeks. Tumor incidence was 1.7% in the MZ-5-156 treated group vs 10% in the control group. Cognition was improved with MZ-5-156 treatment at 12 and 14 mo, but not at 17 mo, as measured by the active avoidance T-maze, the lever press, step-down passive avoidance, and object recognition. Strength, balance, and metabolic parameters were largely unaffected by MZ-5-156. Telomerase activity was improved in heart, aorta, liver, and stomach, but an increase for ileum was not significant. Overall, these findings show positive benefits of treatment with a GHRH antagonist on median life expectancy, tumor incidence, cognition, CNS oxidative stress, and telomerase activity in the aged SAMP8 mouse.[br][br]Session supported by: Ipsen Biopharmaceuticals, Inc.[br][br]Sources of Research Support: VA Merit Review (WAB), NIH R0-1 AG029839 (WAB), the Medical Research Service of the Veterans Affairs Department, South Florida VA Foundation for Research and Education, and the U of Miami Miller School of Medicine, Department of Pathology and Medicine, Division of Hematology/Oncology (all A.V.S.).[br][br]Nothing to Disclose: WAB 2012-06-24T16:45:00 Room 320 2012-06-24T00:00:00 1899-12-30T16:45:00 2465 224 1793 S28-3 T07-S01 Sunday 225 2012


2955 ENDO12L_S29-1 SYMPOSIUM SESSION: BASIC - New Insights into Insulin Action (3:45 PM - 5:15 PM) mTOR Mediates Different Insulin Metabolic Actions Reuben J Shaw Salk Institute for Biological Studies, La Jolla, CA Session supported by: Lilly USA, LLC[br][br]Nothing to Disclose: RJS 2012-06-24T15:45:00 Room 310 2012-06-24T00:00:00 1899-12-30T15:45:00 6100 225 1794 S29-1 T09-S04 Sunday 227 2012


2956 ENDO12L_S29-2 SYMPOSIUM SESSION: BASIC - New Insights into Insulin Action (3:45 PM - 5:15 PM) Hepatic Insulin Signaling Controls Glucose [amp] Lipid Metabolism Morris F White Children[apos]s Hospital, Boston, MA Session supported by: Lilly USA, LLC[br][br]Disclosure Incomplete: MFW 2012-06-24T16:15:00 Room 310 2012-06-24T00:00:00 1899-12-30T16:15:00 6101 225 1795 S29-2 T09-S04 Sunday 228 2012


2957 ENDO12L_S29-3 SYMPOSIUM SESSION: BASIC - New Insights into Insulin Action (3:45 PM - 5:15 PM) Insulin Signaling in Skeletal Muscle: Implications in Type 2 Diabetes Juleen Rae Zierath Karolinska Institutet, Stockholm, Sweden DNA Methylation provides a mechanism by which environmental factors control insulin sensitivity. In this lecture, recent evidence linking obesity and exercise to the control of insulin sensitivity through DNA methylation will be presented. We assessed DNA methylation in skeletal muscle from obese women before and after gastric bypass (GBP). Clinical and molecular changes in obese women were compared against age-matched non-obese women. Obesity was associated with altered expression of a subset of genes enriched in metabolic process and mitochondrial function. After weight loss, the expression of all identified genes was normalized to levels observed in the normal weight woman. Amongst 16 genes analyzed, promoter methylation of eight genes showed a negative association between gene expression and promoter methylation, whereas 5 genes showed a positive association. Using bisulfite sequencing, we further show that promoter methylation of PGC-1[alpha] and PDK4 is altered with obesity and restored to non-obese levels after GBP-induced weight loss. Thus, obesity and GBP-induced weight loss have a dynamic effect on the epigenome. Exercise is the first line of defense against the development of insulin resistance in Type 2 diabetes. Exercise elicits gene expression changes driving structural and metabolic adaptations in skeletal muscle. In the second part of this lecture, evidence for a role of DNA methylation in exercise-induced gene expression will be presented. Whole genome methylation was decreased in skeletal muscle biopsies obtained from 14 young healthy sedentary men and women after an acute bout of exercise. In a separate cohort of eight young healthy sedentary men, exercise induced a dose-dependent expression of PGC-1[alpha], TFAM, PDK4 and PPAR-[delta] together with a marked hypomethylation on each respective promoter. Similarly, promoter methylation of PDK4, PGC-1[alpha], MEF2A and MYOD1 was markedly decreased in mouse soleus muscles 45 min after ex vivo contraction. In L6 myotubes, caffeine induced gene hypomethylation in parallel with an increase in the respective mRNA content. Our results provide evidence that acute gene activation is associated with a dynamic change in DNA methylation in skeletal muscle and suggest that DNA hypomethylation is an early event in contraction-induced gene activation. In conclusion, diet and physical activity are environmental factors that can impact insulin sensitivity through changes in DNA methylation.[br][br]Session supported by: Lilly USA, LLC[br][br]Nothing to Disclose: JRZ 2012-06-24T16:45:00 Room 310 2012-06-24T00:00:00 1899-12-30T16:45:00 2398 225 1796 S29-3 T09-S04 Sunday 229 2012


2958 ENDO12L_S30-1 SYMPOSIUM SESSION: BASIC - Non-Genomic Central Actions of Estrogens (3:45 PM - 5:15 PM) Acute Estrogen Modulation of Synapses in the Hippocampus Catherine Woolley Northwestern University, Evanston, IL Estrogens influence brain function through multiple mechanisms with time courses ranging from minutes to days. Interest in rapid nongenomic estrogen actions has resurged recently, in parallel with recognition that estrogens are produced as neurosteroids in the brains of both males and females. The hippocampus is among the brain regions now known to contain both the enzymatic machinery to synthesize a key estrogen, 17[beta]-estradiol, as well as extranuclear estrogen receptors (ERs) at or near synapses. This suggests that locally synthesized neurosteroid estradiol could acutely regulate synaptic function in the hippocampus in vivo. We have used electrophysiological recording in brain slices from adult rats to investigate mechanisms by which estradiol acutely modulates both excitatory and inhibitory synapses in the hippocampus. Previously, we reported that estradiol rapidly potentiates excitatory synaptic transmission in the CA1 region through an input-specific, ER[beta]-dependent increase in presynaptic glutamate release. More recently, we have found that estradiol also rapidly suppresses inhibitory synaptic transmission in CA1 through an input-specific, ER[alpha]-dependent suppression of GABA release. Further investigations of estradiol-induced suppression of inhibition support a model in which estradiol stimulates an interaction between extranuclear ER[alpha] and postsynaptic mGluR1 to mobilize endocannabinoids that retrogradely suppress GABA release at a subset of inhibitory synapses that contain CB1 receptors. Remarkably, this mechanism appears to be sex-specific, operating in females but not in males. Because estrogens are known to influence multiple hippocampal functions, including learning and memory, affective behaviors, and seizures in epilepsy, understanding the factors that control acute modulation of hippocampal synaptic function by estrogens may suggest new avenues to develop therapies for neurological disorders that involve the hippocampus.[br][br]Sources of Research Support: NIH Grant NS037324 awarded to CSW.[br][br]Disclosure Incomplete: CW 2012-06-24T15:45:00 Room 351 2012-06-24T00:00:00 1899-12-30T15:45:00 2471 226 1797 S30-1 T06-S06 Sunday 231 2012


2959 ENDO12L_S30-2 SYMPOSIUM SESSION: BASIC - Non-Genomic Central Actions of Estrogens (3:45 PM - 5:15 PM) Estradiol Synthesis at the Synapse: A Form of Classic Neuromodulation Barney A Schlinger University of California, Los Angeles, Los Angeles, CA Sex-steroids are recognized for their dramatic impact on brain and behavior. A widely-held perception is that these molecules are derived from peripheral sources and lack the spatial and temporal specificity ascribed to classical neuromodulatory systems. My lab has explored the idea that estradiol can function as a classic neuromodulator, even neurotransmitter, in the vertebrate brain. With a focus on songbirds, I will present evidence for the presence of the estrogen-synthetic enzyme aromatase in pre-synaptic terminals throughout the songbird brain. I will describe experiments demonstrating the acute regulation of pre-synaptic estradiol synthesis in vivo, as well as functional (electrophysiological and behavioral) post-synaptic estradiol actions. Our results meet all the criteria for a neuromodulatory system and shift our perception of estradiol as exclusively a peripheral reproductive signal to a signaling system intrinsic to the brain itself. We apply the term [ldquo]synaptocrine[rdquo] to describe this form of neuromodulation (Saldanha et al., 2011).[br][br]Saldanha, C.J., Remage-Healey, L., Schlinger, B.A. 2011. Synaptocrine Signaling: steroid synthesis and action at the synapse. Endocrine Revs. 32:532 [ndash] 549.[br][br]Sources of Research Support: NIMH Grant 061994.[br][br]Nothing to Disclose: BAS 2012-06-24T16:15:00 Room 351 2012-06-24T00:00:00 1899-12-30T16:15:00 381 226 1798 S30-2 T06-S06 Sunday 232 2012


2960 ENDO12L_S30-3 SYMPOSIUM SESSION: BASIC - Non-Genomic Central Actions of Estrogens (3:45 PM - 5:15 PM) Post-Ischemic Administration of Estradiol [amp] Analogs That Bind to Membrane Receptors Promotes Survival of CA1 Hippocampal Neurons in Middle-Aged Rats Anne M Etgen Albert Einstein College of Medicine, Bronx, NY Considerable evidence indicates that 17[beta]-estradiol (17[beta]-E2) is neuroprotective in animal models of focal and global ischemia. We recently showed that acute administration of 17[beta]-E2 and certain estrogenic compounds that do not bind classical estrogen receptors (ER), such as G1 and STX, also reduce hippocampal neuronal loss in middle-aged rats (Lebesgue et al. 5:e8642, 2010). Thus, the neuroprotective actions of estrogens are retained in older animals, and these effects may be mediated via membrane-associated receptors. We now report on the neuroprotective effects of post-ischemic administration of several estrogen analogs, including 17[alpha]-E2 and [italic]ent[/italic]-17[beta]-E2 (mirror image of 17[beta]-E2, which does not bind ERs) in middle-aged rats subjected to long-term hormone withdrawal. Two or 6 months after ovariectomy, animals underwent 10 min transient global ischemia. A single infusion of 17[beta]-E2, [italic]ent[/italic]-17[beta]-E2, 17[alpha]-E2, estrone or other estrogen analogs was administered into the lateral ventricle immediately after reperfusion. Infusion of 17[beta]-E2 (2.25 [micro]g) rescued 40-60% of CA1 pyramidal neurons in rats that were hormone-deprived for either 2 or 6 months. A high dose of [italic]ent[/italic]-17[beta]-E2 (50 [mu]g) was also neuroprotective (50-65% of neurons survived). However, 17[alpha]-E2, estrone, [italic]ent[/italic]-estrone and a variety of other estrogen analogs did not reduce cell death. To determine whether 17[beta]-E2 and other neuroprotective estrogen analogs have similar functional properties in synaptic transmission and plasticity in the hippocampal neurons, we measured field excitatory postsynaptic potentials, paired pulse facilitation and long-term potentiation (LTP) generated in the CA1 pyramidal cell layer by Schaffer collateral stimulation. Our electrophysiological data suggest that 17[beta]-E2, G1 and STX, but not other estrogens, facilitate synaptic responses in CA1 pyramidal neurons. Moreover, bath application of 1 nM 17[beta]-E2 onto brain slices derived from aged females after 6 months hormone deprivation significantly enhanced excitatory transmission at CA1 synapses evoked by Schaffer collateral stimulation, and normal LTP was induced in response to high frequency stimulation. The magnitude of the enhancement was indistinguishable from that recorded in slices from young rats. Our findings suggest that the neuroprotective effects of estrogens given after ischemia may be mediated through membrane-initiated action and that the aging brain remains responsive to these actions even after a long period of hormone withdrawal.[br][br]T. Inagaki, Albert Einstein Coll. Med. P.E.Castillo, Albert Einstein Coll. Med. D. Covey, Washington Univ. N. Kaneko, Albert Einstein Coll. Med. R.S. Zukin, Albert Einstein Coll. Med.[br][br]Sources of Research Support: NIH Grant R01 AG027702 to AE.[br][br]Nothing to Disclose: AME 2012-06-24T16:45:00 Room 351 2012-06-24T00:00:00 1899-12-30T16:45:00 2460 226 1799 S30-3 T06-S06 Sunday 233 2012


2961 ENDO12L_S31-1 SYMPOSIUM SESSION: TRANSLATIONAL - Novel Therapeutic Approaches to Metastatic Thyroid Cancer (Spanish) (3:45 PM - 5:15 PM) TGF[beta] Signaling Pathway in Thyroid Cancer Progression: A Potential Therapeutic Target Garcilaso Riesco-Eizaguirre Hospital University Ersitario La Paz, Madrid, Spain In addition to the well-established genetic events identified in thyroid cancer, signalling through growth factors and their receptors is considered essential for cancer progression. Some of these growth factors have been identified as modifiers of the behaviour of transformed thyroid cells. Among them, TGFbeta is becoming increasingly important as an oncogenic factor that cooperates with other genetic events and/or signalling pathways in promoting progression and dedifferentiation in thyroid cancer. The BRAF oncogene mediates signal transduction through the MEK-ERK pathway and is one of the most important genetic events in papillary thyroid carcinomas. It is associated with poorer clinicopathological outcomes and independently predicts recurrences, particularly those that have lost the ability to accumulate iodide, a property mediated by the sodium iodide symporter (NIS). The loss of NIS expression is one of the most significant hallmarks of advanced thyroid cancer, an event that leads to radioiodide-refractory metastatic disease and spells a markedly worse outcome for the affected patients. BRAF-positive tumors exhibit a significantly reduced NIS expression and BRAF represses NIS functional expression in thyroid cells grown in vitro. The mechanism by which BRAF induces NIS repression is based on the operation of an autocrine TGF[beta] loop: BRAF activation induces TGF-[beta] secretion which in turn downregulates Pax8 and evokes a Smad3-dependent inhibition of Pax8 binding to the NIS promoter. Additionally, TGFbeta in cooperation with the MEK-ERK pathway, promotes epithelial to mesenchimal transition (EMT) and tumor invasion. In human thyroid cancers, a high TGFbeta-Smad activity is present in invasive tumors and is associated with a poor clinical outcome of the patients. The majority of human thyroid cancer cell lines have lost the cytostatic response to TGFbeta and small-molecule TGFbeta inhibitors are able to suppress cell proliferation and cell invasion. Interestingly, TGFbeta inhibition mainly impairs single cell motility but not the collective movement of the cells. Single cell motility has been involved in the genesis of blood-borne distant metastases. Finally, analysis of a large panel of EMT-related genes revealed two novel factors that are regulated by TGFbeta that may be playing an important role in thyroid carcinogenesis. In conclusion, TGF[beta] has a crucial role in thyroid cancer progression being a molecular marker of poor prognosis.[br][br]Nothing to Disclose: GR-E 2012-06-24T15:45:00 Theater C 2012-06-24T00:00:00 1899-12-30T15:45:00 2521 227 1800 S31-1 T02-S11 Sunday 235 2012


2962 ENDO12L_S31-2 SYMPOSIUM SESSION: TRANSLATIONAL - Novel Therapeutic Approaches to Metastatic Thyroid Cancer (Spanish) (3:45 PM - 5:15 PM) Targeting MAPK Signaling: Promises [amp] Challenges James A Fagin Memorial Sloan-Kettering Cancer Center, New York, NY Disclosure Incomplete: JAF 2012-06-24T16:15:00 Theater C 2012-06-24T00:00:00 1899-12-30T16:15:00 6030 227 1801 S31-2 T02-S11 Sunday 236 2012


2963 ENDO12L_S31-3 SYMPOSIUM SESSION: TRANSLATIONAL - Novel Therapeutic Approaches to Metastatic Thyroid Cancer (Spanish) (3:45 PM - 5:15 PM) New Therapies [amp] Clinical Trials in Medullary [amp] Differentiated Thyroid Cancer Nelson Wohllk Universidad de Chile, Santiago, Chile Las opciones para el tratamiento del Cancer diferenciado (CDT) o Medular (CMT) incluyen el uso de radioyodo, reduccion quirurgica dirigida, radioterapia y quimioterapia; estas ultimas opciones con poco exito. CDT metastasico concentra pobremente el radioyodo en la medida que se hace menos diferenciado. Pacientes con CMT o CDT que progresan podrian ser enrolados en ensayos clinicos (EC) en curso con uso de inhibidores de quinasas (1). En EC fase III, el objetivo principal es generalmente comparar el efecfo de la droga y el placebo sobre sobrevida libre de progresion (PFS), de acuerdo con el criterio de RECIST (2). Vandetanib, un inhibidor de las senales quinasas RET, VEGFR y EGFR ha mostrado previamente actividad antitumoral en estudios de fase II en pacientes con CMT hereditario progresivo. En CMT existe uno solo EC en fase III que ha sido completado. Este EC mostro que la PFS mediana[br]fue 11 meses mas larga que en el grupo asignado aleatoriamente a vandetanib y que la tasa de respuesta objetiva fue de 45% (3). Vandetanib fue aprobado por la FDA en Abril de 2011. Cabozantinib (XL184) es potente inhibidor dual de las vias MET y VEFG disenado para bloquer el escape tumoral. Un EC en curso en fase III en pacientes con CMT conocido como EXAM, cumplio su primer objetivo final, mostrando una mejoria de 7,2 meses en la sobrevida promedio en pacientes con cabozantinib vs placebo (4). En CDT hay una droga en fase III reclutando pacientes: E7080(Lenvatinib) en CDT refractario a 131I. Un EC en fase II estudio el efeco de lenvatnib (cuyos blancos son VEGFR1-3, FGFR 1-4, RET, cKit and PDGFRBeta) en 58 pacientes y la PFS mediana fue de 12.6 meses (5). Sorafenib (inhibidor de VEGFR2, VEGFR3, RET y BRAF) y sunitinib (inhibidor de los 3VEGFRs, RET, y RET/PTC subtypes 1 and 3) han sido aprobados por la FDA y la EMEA en otros tumores, estando asi potencialmente disponibles para su uso no oficial en pacientes con CDT y CMT metastasico. Ambas drogas han tenido efecto mas biende estabilizacion de la enfermedad (6). Para Sorafenib existe EC en fase III en curso en CDT. Pazopanib es inhibidor multiquinasa con EC fase III en curso. Existen otros EC en fase II usando sorafenib junto a everolimus o tenserolimus, etc. (7). Eventos adversos para cualquier inhibidors de las quinasas ocurren frecuentmente siendo frecuente la diarrea, erupcion cutanea, hipertension, prolongacion de QTc, dermatitis aceneiforme/acne, fatiga e hipotiroidismo.[br][br]1.-Gild, M. L. et al. Nat. Rev. Endocrinol 2011 Aug 23;7:617-24. 2.-Therasse P. J Natl Cancer Inst 2000; 92:205-16. 3.-Wells SA Jr, et al. J Clin Oncol 2011; 28:767-772. 4.-http://www.exelixis.com/cabozantinib/clinical-trial-information/medullary-thyroidcancer. 5.-Sherman, S. I. et al. A phase II trial of the multitargeted kinase inhibitor E7080 in. advanced radioiodine (RAI)-refractory differentiated thyroid cancer (DTC) [abstract]. J. Clin. Oncol. 29, a5503 (2011). 6.-Cabanillas, M. E. et al. J. Clin. Endocrinol. Metab. 95, 2588-2595 (2010). 7.- http://www.clinicaltrials.gov/.[br][br]Nothing to Disclose: NW 2012-06-24T16:45:00 Theater C 2012-06-24T00:00:00 1899-12-30T16:45:00 2510 227 1802 S31-3 T02-S11 Sunday 237 2012


2964 ENDO12L_S32-1 SYMPOSIUM SESSION: CLINICAL - Origins [amp] Markers of the Polycystic Ovarian Syndrome (PCOS) (3:45 PM - 5:15 PM) Genome-Wide Association Studies of PCOS Joop S E Laven Erasmus MC, University Medical Center, Rotterdam, Netherlands PCOS is the commonest endocrine abnormality in women of reproductive age. It represents the major cause of anovulatory infertility and is also associated with androgen excess, hirsutism and acne[sup]1-3[/sup]. The typical biochemical features are elevated serum levels of testosterone and luteinizing hormone (LH) along with characteristic metabolic disturbances including insulin resistance and abnormalities of energy expenditure[sup]4,5[/sup]. Crucially, PCOS is now recognised as a major risk factor for the development of type 2 diabetes (TD2) and cardiovascular disease in later life[sup]6-8[/sup]. At least in part, this reflects the strong associations between PCOS and obesity, with the latter being an amplifier of PCOS[sup]9,10[/sup].[br]The aetiology of PCOS is unclear but exposure to excess androgen during intrauterine life in animals results in many of the features of human PCOS[sup]11,12[/sup]. Hence, PCOS is a genetically-determined disorder of ovarian function leading to hyperandrogenism both during fetal life and subsequently during puberty[sup]13,14[/sup]. Due to it[apos]s heterogeneous nature, historical disagreements about the definitions and the fact that the syndrome is restricted to women of reproductive age did hamper genetic family studies[sup]15-18[/sup]. There is familial clustering of cases and concordance of symptoms in identical twins compared with non-identical twins[sup]15,16,18[/sup]. Moreover, there is evidence for heritability of endocrine and metabolic features of PCOS[sup]16,19[/sup]. The mode of inheritance is complex and multifactorial[sup]15,20-22[/sup]. The search for genetic variants influencing PCOS susceptibility yielded few results identifying SNP[apos]s in the FSH, LH and AMH Receptor genes which were associated with the PCOS phenotype[sup]23,24[/sup]. Other studies in PCOS have dispelled previous claims of association overlap between T2D-susceptibility variants and PCOS risk[sup]25-27[/sup]. Most recently variants within the [italic]FTO [/italic]gene had a detectable impact on PCOS risk. However, in a replication study the BMI susceptibility gene distribution did not differ between controls and women with PCOS.[br]Finally, recent genome-wide association studies (GWAS) of PCOS have identified single nucleotide poplymorphisms (SNP[apos]s) in 3 loci in Chinese women with PCOS. Several genes are close to these newly identified loci such as the [italic]LHCGR[/italic] and the [italic]FSHR[/italic] genes. Currently, GWAS efforts are underway in Europe as well as in the United States and they will eventually provide evidence for similar and additional loci of interest that are associated with PCOS[sup]28,29[/sup].[br][br]1. Franks S, N Engl J Med 1995; 333:853. 2. Knochenhauer E et al., J Clin Endocrinol Metab 1998; 83:3078. 3. Ehrmann DA 2005 Polycystic ovary syndrome. N Engl J Med 352:1223-1236. 4. Robinson S et al., Clin Endocrinol (Oxf) 1992; 36:537. 5. Dunaif A. Endocr Rev 1997; 18:774. 6. Dahlgren E et al., Fertil Steril 1992; 57:505. 7. Wild S et al., Hum Fertil (Camb) 2000, 3:101. 8. Solomon CG et al., J Clin Endocrinol Metab 2002; 87:2013. 9. Legro RS et al., J Clin Endocrinol Metab 1999; 84:165. 10. Ehrmann DA et al., Diabetes Care1999;22:141. 11. Dumesic DA et al. Fertil Steril 1998;70:94. 12. Eisner JR et al., J Clin Endocrinol Metab 2000; 85:1206. 13. Abbott DH etal., J Endocrinol 2002; 174:1. 14. Franks S et al., Int J Androl 2002; 29:278. 15. Franks S et al., Reprod 1997; 12:2641. 16. Legro RS et al., Proc Natl Acad Sci U S A 1998; 95:14956. 17. Franks S et al., Rev Endocr Metab Disord 2004;5:69. 18. Vink JM et al., J Clin Endocrinol Metab 2005;91:2100. 19. Legro RS et al., J Clin Endocrinol Metab 2002;87:2128. 20. Hague WM et al., Clin Endocrinol (Oxf) 1988;29:593. 21. Simpson JL Oxford: Blackwell Scientific Publications 1992; 59. 22. Stewart DR et al., J Clin Endocrinol Metab 2006;91:4112. 23. Laven JS et al., Fertil Steril 2003;80:986. 24. Kevenaar ME et al., Hum Reprod 2007;22:1547. 25. Gaasenbeek M et al., J Clin Endocrinol Metab 2004;89:2408. 26. Powell BL et al., J Clin Endocrinol Metab. 2005;90:2988. 27. Ruokonen A et al. J Clin Endocrinol Metab 2005;90:2988. 28. Chen ZJ et al., Nat Genet. 2011;43:55. 29. Urbanek M. Nat Clin Pract Endocrinol Metab. 2007;3:103.[br][br]Sources of Research Support: None.[br][br]Nothing to Disclose: JSEL 2012-06-24T15:45:00 Theater B 2012-06-24T00:00:00 1899-12-30T15:45:00 2432 228 1803 S32-1 T05-S02 Sunday 239 2012


2965 ENDO12L_S32-2 SYMPOSIUM SESSION: CLINICAL - Origins [amp] Markers of the Polycystic Ovarian Syndrome (PCOS) (3:45 PM - 5:15 PM) Early Origins of PCOS Stephen Franks Imperial College London, London, UK Polycystic ovary syndrome (PCOS) frequently presents during adolescence and is the commonest cause of menstrual irregularity and hirsutism. The characteristic endocrine abnormalities include hypersecretion of androgens and LH. Metabolic dysfunction is also a feature of many young women with PCOS. Hyperinsulinemia and insulin resistance, which can be regarded as an exaggeration of the normal metabolic changes that occur during puberty, are further amplified by obesity. It is of concern that adults with symptoms of PCOS who have the most unfavorable metabolic profile are those who were obese as children. The etiology of PCOS remains uncertain but there is evidence for a primary abnormality of ovarian androgen production which is manifest at puberty but may have its origins in childhood or even during fetal development. We have proposed that polycystic ovary syndrome does indeed originate in fetal life (1,2). This hypothesis is based on data from animal models and is supported by clinical studies. Animal models include Rhesus monkeys or sheep that have been exposed prenatally to high doses of androgen (1,3), and rodents given androgens before or during puberty (4,5). It is suggested that, in human females, exposure to excess androgen, at any stage from fetal development of the ovary to the onset of puberty, leads to many of the characteristic features of PCOS, including abnormalities of LH secretion and insulin resistance. We hypothesize that, in humans with PCOS, the development of the PCOS phenotype results primarily from a genetic predisposition for the fetal ovary to hypersecrete androgen (1,2) but despite strong evidence for a major genetic contribution to the etiology of PCOS, the key gene (or more probably genes) involved remain to be clearly defined. At present, it is unclear whether the maternal environment directly influences the development of PCOS in the offspring. Maternal androgen excess is unlikely to affect the fetus, because the placenta presents an effective barrier, but metabolic disturbances during pregnancy could affect development of the syndrome in the fetus. In postnatal life the natural history of PCOS can be further modified by factors affecting insulin secretion and/or action, most importantly, nutritional.[br][br](1) Abbott DH, Dumesic DA, Franks S Developmental origin of polycystic ovary syndrome- a hypothesis. J Endocrinol. 2002 174:1-5. (2) Franks S, McCarthy MI, Hardy K. Development of polycystic ovary syndrome: involvement of genetic and environmental factors. Int J Androl. 2006 29:278-85; Review. (3) Dumesic DA, Abbott DH, Padmanabhan V. Polycystic ovary syndrome and its developmental origins. Rev Endocr Metab Disord. 2007 8:127-41; Review. (4) Tyndall V, Broyde M, Sharpe R, Welsh M, Drake AJ, McNeilly AS. Effect of androgen treatment during foetal and/or neonatal life on ovarian function in prepubertal and adult rats. Reproduction. 2012 143:21-33. (5) van Houten EL, Kramer P, McLuskey A, Karels B, Themmen AP, Visser JA. Reproductive and metabolic phenotype of a mouse model of PCOS. Endocrinology. 2012 Feb 14. [Epub ahead of print].[br][br]Sources of Research Support: Medical Research Council (UK) Programme Grant G0802782 awarded to SF.[br][br]Nothing to Disclose: SF 2012-06-24T16:15:00 Theater B 2012-06-24T00:00:00 1899-12-30T16:15:00 2414 228 1804 S32-2 T05-S02 Sunday 240 2012


2966 ENDO12L_S32-3 SYMPOSIUM SESSION: CLINICAL - Origins [amp] Markers of the Polycystic Ovarian Syndrome (PCOS) (3:45 PM - 5:15 PM) Ovarian Morphology in PCOS: A Critical Reappraisal of the Use of Ultrasonagraphy Judith Mary Adams Massachusetts General Hospital, Boston, MA Polycystic ovary syndrome, characterized by menstrual irregularities and hyperandrogenism is associated with a pathological appearance of the ovary. In 1985, we defined polycystic ovaries (PCOM) as the presence of [underline][gt][/underline]10cysts, arranged peripherally around a dense core of stroma. Despite the apparent importance of PCOM in PCOS, the diagnostic criteria for PCOS defined by the 1990 NIH workshop did not include ovarian morphology. A revised definition of PCOS proposed in 2003 at a consensus workshop in Rotterdam reemphasized the importance of PCOM in the diagnostic criteria, requiring a follicle number of 12 or more in either ovary and /or an ovarian volume of more than 10ml. The Rotterdam classification is controversial, since it broadens the spectrum of PCOS, incorporating women with hyperandrogenism and PCOM without menstrual dysfunction, or PCOM, oligomenorrhea, without hyperandrogenism. The Rotterdam prevalence estimates of PCOS are up to twice those obtained with the NIH criteria, raising the question of whether this is a more accurate reflection of the true prevalence of the disorder, or a gross overestimation. Recent studies from Denmark and Holland have shown a prevalence of PCOM in up to 80% of young healthy volunteers. This prevalence decreased with age. Our longitudinal and cross sectional studies of women with PCOS showed a decrease in ovarian volume and follicle number with age, emphasizing the need for age-based criteria to make the diagnosis.[br]It has been suggested that the follicle number and ovarian volume criteria be revised. Several studies have suggested the incorporation of serum AMH levels as an additional diagnostic marker. 3D ultrasound allows quantitative assessment of ovarian volume, blood flow and stromal echogenicity, and as technology advances, may become more widely available in a clinical setting.[br][br]Supported by NIH Grant U01 HD 4417; National Center for Research Resources General Clinical Research Centers Program Grant M01-RR-01066; 1R01HD065029. Nothing to Disclose: JMA.[br][br]Nothing to Disclose: JMA 2012-06-24T16:45:00 Theater B 2012-06-24T00:00:00 1899-12-30T16:45:00 2529 228 1805 S32-3 T05-S02 Sunday 241 2012


2967 ENDO12L_S33-1 SYMPOSIUM SESSION: TRANSLATIONAL - The Androgen Receptor: Eveready Battery of Prostate Cancer (3:45 PM - 5:15 PM) AR Splice Variants in Prostate Cancer Progression Scott M Dehm Masonic Cancer Center, Minneapolis, MN Prostate cancer is the most frequently diagnosed male cancer and second leading cause of male cancer deaths. The androgen receptor (AR) transcription factor is a master regulator of the prostate lineage during normal development, and AR activity continues to support prostate gland function in healthy adults. Importantly, the AR retains this master regulator function in prostate cancer cells. Therefore, blocking the production or action of androgens via androgen depletion therapy (ADT) is an effective systemic targeted therapy for locally advanced or metastatic disease. Invariably, a lethal castration-resistant prostate cancer phenotype emerges through various mechanisms of aberrant AR re-activation during ADT. One mechanism of ADT-resistance that has recently been revealed by our group and others is the synthesis of truncated AR variants lacking the canonical steroid receptor ligand-binding domain (LBD). Truncated AR variants arise through alterations in AR mRNA splicing, and various discrete species have been identified in prostate cancer cell lines, xenografts, and clinical specimens. Most commonly, these species consist of the AR NH[sub]2[/sub]-terminal transcriptional activation domain (NTD) and the DNA-binding domain (DBD) but have short, variable COOH-terminal extensions instead of an AR LBD. Through high-resolution copy number analysis and next-generation paired-end re-sequencing of the 180kb AR gene in models of castration-resistant prostate cancer, we have identified diverse focal AR gene rearrangements that underlie altered splicing patterns and high-level synthesis of truncated AR variants. Functionally, we have found that the AR NTD/DBD core is sufficient for truncated AR variants to access the nucleus, activate constitutive, ligand-independent expression of AR target genes, and support androgen-independent growth of prostate cancer cells. In summary, AR gene rearrangement and synthesis of truncated, constitutively active AR splice variants represent a novel mechanism by which prostate cancer cells are able to satisfy their ongoing lineage addiction to AR activity, and thereby continue to grow during ADT.[br][br]Sources of Research Support: Prostate Cancer Foundation Young Investigator Award; U.S. Department of Defense Prostate Cancer Research Program New Investigator Award W81XWH-10-1-0353; American Cancer Society Research Scholar Grant RSG-12-031-01-TBE.[br][br]Nothing to Disclose: SMD 2012-06-24T15:45:00 Room 362 2012-06-24T00:00:00 1899-12-30T15:45:00 2500 229 1806 S33-1 T02-S06 Sunday 243 2012


2968 ENDO12L_S33-2 SYMPOSIUM SESSION: TRANSLATIONAL - The Androgen Receptor: Eveready Battery of Prostate Cancer (3:45 PM - 5:15 PM) Androgenic Pathways in Prostate Cancer Progression Alice C Levine Mt Sinai Medical Center, New York, NY Prostate cancer (PCa) preferentially metastasizes to bone with a resulting mortality of approximately 70%. The metastases are associated with crippling complications including severe pain, fractures, spinal cord compression and bone marrow suppression. Unfortunately, there are currently no effective therapies for the prevention or treatment of PCa bone metastases. Androgens are essential for normal prostate development and are necessary but not sufficient for the development of prostate cancer. Androgen deprivation therapy (ADT) has long been the mainstay of treatment for PCa bone metastases, providing palliation of symptoms in the majority of patients, followed by relapse and progression, which is still driven by androgens and AR-signaling in castrate-resistant disease in the majority of cases. In spite of these demonstrated effects of androgens on the early and very late stages of PCa, less is known about androgenic effects during disease progression to incurable bone metastases. The majority of published preclinical studies demonstrate a stimulatory effect of androgens and androgen receptor (AR) signaling on the multistep process of PCa bone metastases, including androgenic promotion of local PCa growth, angiogenesis, invasion, bone-targeting, and enhancement of Wnt-signaling in osteoblasts. However, in spite of these demonstrated effects on PCa progression and bone metastases, the only proven survival benefit of early androgen deprivation therapy (ADT) is as neo-adjuvant treatment in men with high-risk, advanced localized disease receiving external beam radiotherapy. The lack of efficacy of early ADT in other settings is likely due to the profound effects of androgen deprivation on normal bone cells; ADT promotes osteoclastogenesis which may provide an entry for PCa cells into the bone and give them access to the rich storehouse of growth factors in the bone matrix.[br][br]Nothing to Disclose: ACL 2012-06-24T16:15:00 Room 362 2012-06-24T00:00:00 1899-12-30T16:15:00 2495 229 1807 S33-2 T02-S06 Sunday 244 2012


2969 ENDO12L_S33-3 SYMPOSIUM SESSION: TRANSLATIONAL - The Androgen Receptor: Eveready Battery of Prostate Cancer (3:45 PM - 5:15 PM) Novel Therapeutic Targeting Androgenic Pathways in Prostate Cancer Chris Logothetis MD Anderson Cancer Center, Houston, TX Disclosure Incomplete: CL 2012-06-24T16:45:00 Room 362 2012-06-24T00:00:00 1899-12-30T16:45:00 6024 229 1808 S33-3 T02-S06 Sunday 245 2012


2970 ENDO12L_S34-1 SYMPOSIUM SESSION: TRANSLATIONAL - Update in Endocrine Hypertension (3:45 PM - 5:15 PM) How Much Do We Gain by Measurement of Cortisol Metabolites Produced by Aldosterone Synthase? The Noninvasive Method Determining Aldosteronoma with a High Specificity Fumitoshi Satoh Tohoku University Hospital, Sendai, Miyagi, Japan Adrenal venous sampling (AVS) is a gold standard for subtype differentiation in primary aldosteronism (PA), but is an invasive examination and needs experienced radiologists. AVS has sometimes been criticized for its less standardization of techniques and protocols even if there have been international collaboration works (1). 18-oxocortisol (18-oxoF) is a derivative of cortisol (F) that is produced by aldosterone synthase (CYP11B2), and its clinical usefulness has not been clarified. We demonstrated that 18-oxoF levels were higher in the adrenal vein at the side of aldosteronoma compared with bilateral hyperplasia (2). We prospectively measured 18-oxoF, using highly sensitive liquid chromatography tandem mass spectrometry (LC/MS/MS), in peripheral plasma of patients with PA who underwent AVS to differentiate aldosterone producing adenoma (APA) from bilateral hyperplasia as previous reports (3). The study was performed in 116 patients with surgically-proven APA and 121 patients with bilateral hyperplasia diagnosed by successful AVS. The levels of 18-oxoF were significantly higher in the peripheral plasma from the patients with APA than those from the patients with bilateral hyperaldosteronism (BHA) diagnosed by AVS. The cutoff value (4.7 ng/dl) of 18-oxoF levels was calculated by ROC analysis of APA versus bilateral hyperplasia with sensitivity of 0.80 and specificity of 0.97. In 114 cases with CT-positive unilateral adrenal masses, the cutoff value (4.6 ng/dl) of 18-oxoF levels was calculated with sensitivity of 0.80 and specificity of 0.97. In these cases, the 18-oxoF levels in 60 of 87 cases (80%) with APA were higher than 4.6 ng/dl, and while, those in 29 of 30 cases (97%) with bilateral hyperplasia were lower than 4.6 ng/dl. According to peripheral blood levels of 18-oxoF, 80% of APA patients with unilateral adrenal masses might be able to undergo ipsilateral adrenalectomy without AVS. Moreover, ROC analysis of APA versus primary hypertension demonstrated that APA cases were diagnosed by 18-oxoF with sensitivity of 0.80 and specificity of 0.94. Thus, peripheral blood levels of 18-oxoF might be a clinically useful biomarker for differentiating APA from bilateral hyperplasia or primary hypertension.[br][br]1) Satoh F et al. Hypertension. 2012 Feb 13 on line. 2) Satoh F et al. J Clin Endocrinol Metab. 2011; 96(8). 3) Satoh F et al. Hypertens Res. 2007; 30(11).[br][br]Nothing to Disclose: FS 2012-06-24T15:45:00 Theater A 2012-06-24T00:00:00 1899-12-30T15:45:00 2491 230 1809 S34-1 T01-S05 Sunday 247 2012


2971 ENDO12L_S34-2 SYMPOSIUM SESSION: TRANSLATIONAL - Update in Endocrine Hypertension (3:45 PM - 5:15 PM) New Gene Mutations in Pheochromocytomas Massimo Mannelli University of Florence, Florence, Italy Pheochromocytomas (Pheos) and Paragangliomas (PGLs) are neural crest-derived tumors arising in the adrenal medulla or in the sympathetic or parasympathetic paraganglia. Sympathetic PGLs are extraadrenal abdominal/thoracic catecholamine secreting tumors (sPGLs) while parasympathetic PGLs occur in the head and neck region (HNPGL). Pheos and PGLs can occur as sporadic or hereditary. At present, 35-40% of these tumors are due to germ-line mutations in susceptibility genes (1), some of which have recently been discovered.[br]In addition to the [Prime]old known[Prime] [italic]VHL, RET[/italic]and [italic]NF1[/italic] genes, responsible for von Hippel-Lindau, MEN2 and Neurofibromatosis type 1 syndromes, other susceptibility genes have been discovered in the last 10 years. Some of them are involved in succinate-dehydrogenase (SDH) or mitochondrial complex II function and assembling. They are the four nuclear genes encoding the four SDH subunits ([italic]SDHA, SDHB, SDHC[/italic] and [italic]SDHD[/italic]) and the gene responsible for SDHA flavination ([italic]SDHAF2[/italic]). Among these, the most often mutated genes are [italic]SDHD[/italic] and [italic]SDHB[/italic] responsible for syndromes named PGL1 and PGL4, respectively, the first characterized by multiple HNPGLs, sometimes associated to sPGLs and the second by abdominal sPGLs presenting as malignant in high percentage (30-50%) and sometimes associated to renal cell carcinoma (2). Recently, two additional susceptibility genes have been discovered: [italic]TMEM127[/italic] (3) and [italic]MAX[/italic] (4), both causing Pheos, often bilateral. Transcriptional analyses of Pheo/PGL have shown that these tumors can be divided into two distinct clusters (5). Cluster 1 includes tumors affecting [italic]VHL[/italic] and [italic]SDH[/italic] mutation carriers and exhibiting a pseudohypoxia phenotype; Cluster 2 includes tumors affecting [italic]RET, NF1, MAX[/italic] and [italic]TMEM127[/italic] mutation carriers and exhibiting a RAS/RAF/MAPK signaling phenotype.[br]Two other genes have been associated to Pheos/PGL: [italic]KIF1B[beta][/italic], a tumor suppressor gene necessary for neuronal apoptosis, whose germ-line mutations have been found in two patients affected by bilateral Pheos (6) and [italic]EGLN1[/italic] a gene encoding a prolyl hydroxylase involved in HIF-[alpha] degradation, whose germ-line mutation has been found in a patient affected by erytrocytosis and recurrent PGLs (7).[br]Why mutations in so many different genes lead to the formation of similar tumors and why neural crest-derived tissues are preferentially affected by these gene mutations are still unanswered questions. The complete mechanisms leading from gene mutations to Pheo/PGL development are still undiscovered.[br][br]1) Mannelli M et al., J Clin Endocrinol Metab 2009;94:1541. 2) Gimenez-Roqueplo AP et al., Cancer Res 2003; 63: 5615. 3) Qin Y et al., Nat Genet 2010; 42:229. 4) Comino-Mendez I et al., Nat Genet 2011; 43:663. 5) Burnichon N et al., Hum Mol Genet 2011; 20:3974. 6) Yeh IT et al., Hum Genet 2008; 124:279. 7) Ladroue C et al., N Engl J Med 2008; 359: 2685.[br][br]Nothing to Disclose: MM 2012-06-24T16:15:00 Theater A 2012-06-24T00:00:00 1899-12-30T16:15:00 2393 230 1810 S34-2 T01-S05 Sunday 248 2012


2972 ENDO12L_S34-3 SYMPOSIUM SESSION: TRANSLATIONAL - Update in Endocrine Hypertension (3:45 PM - 5:15 PM) Interaction of Aldosterone [amp] Salt on Cardiac Remodeling Michael Stowasser University of Queensland Department of Medicine, Brisbane, Australia Nothing to Disclose: MS 2012-06-24T16:45:00 Theater A 2012-06-24T00:00:00 1899-12-30T16:45:00 6008 230 1811 S34-3 T01-S05 Sunday 249 2012


2973 ENDO12L_NS7 ENDOCRINE NURSES SYMPOSIA: CLINICAL - Addison[apos]s Disease (3:15:00 PM - 4:00 PM) Addison[apos]s Disease Baha M Arafah Case Western Reserve University, Cleveland, OH Nothing to Disclose: BMA 2012-06-24T16:00:00 Hilton Americas-Houston, Ballroom of the Americas BC 2012-06-24T00:00:00 1899-12-30T16:00:00 6248 220 1781 NS7 ENS7 Sunday 250 2012


2974 ENDO12L_L4-1 PLENARY: TRANSLATIONAL - Fukushima Daiichi Nuclear Disaster [amp] Radiation Health Risk Management (5:30 PM - 6:30 PM) Fukushima Daiichi Nuclear Disaster and Radiation Health Risk Management Shunichi Yamashita Nagasaki University, Nagasaki, Japan Fukushima Daiichi Nuclear Accident followed a multidimensional disaster that combined to destroy the local infrastructure on which the safety system depended. Countermeasures including evacuation, sheltering, and control of the food chain were implemented in a timely manner. To date there have been no acute radiation injuries from the nuclear accident. Stable iodine was not generally administered to the public. For all that, according to the reported monitoring results, the thyroid doses were low. Taking these factors into account, together with the magnitude of the reported levels of radioactive substances released into the atmosphere and the ocean, the radiation-related physical health consequences on the general public, including evacuees, is likely to be limited and much lower than that from Chernobyl, where the only conclusive short- and mid-term radiation-induced health effect in the population was thyroid cancer in children drinking milk contaminated with high levels of radioactive iodine. However, the social, psychological, and economic impact of the Fukushima nuclear accident is expected to be considerable. Because of these impacts, continued monitoring and characterization of the levels of radioactivity in the environment are vital for obtaining the informed consent to the decisions on various issues such as the extent to which populations can return to their homes. Now we are handling the official plans of the Fukushima Health Management Survey, which includes the basic survey and four detailed surveys: thyroid ultrasound examination, comprehensive health check-up, mental health and life-style survey, and pregnancy and child survey, to prospectively take care of all the residents in Fukushima for a long time. Also we need to realize the importance of radiation biology and stochastic effects of low-dose radiation exposure. Japan has considerable expertise in radiation related issues but transparency in risk assessment and decision-making and a reliable relationship with the public are urgent. The Japanese Government and the international organizations should, therefore, decide how to benefit best from the lessons learned from Chernobyl and Fukushima so that we will be able to effectively continue stronger cooperation in the long term. To overcome many difficulties existing in Fukushima just one year after the accident, the current problems and future planning in Fukushima will be presented at the standpoint of radiation health risk management.[br][br]Nothing to Disclose: SY 2012-06-24T17:30:00 Grand Ballroom AB 2012-06-24T00:00:00 1899-12-30T17:30:00 2415 244 1827 L4-1 PL04-1 Sunday 251 2012


2975 ENDO12L_L4-2 PLENARY: TRANSLATIONAL - Genetic Disorders of Thyroid Hormone Synthesis [amp] Action (5:30 PM - 6:30 PM) Genetic Disorders of Thyroid Hormone Synthesis [amp] Action V Krishna K Chatterjee University of Cambridge/Addenbrooke[apos]s Hospital, Cambridge, UK Nothing to Disclose: VKKC 2012-06-24T17:30:00 Grand Ballroom AB 2012-06-24T00:00:00 1899-12-30T17:30:00 6125 245 1828 L4-2 PL04-2 Sunday 252 2012


2976 ENDO12L_L5-1 PLENARY: TRANSLATIONAL - Roy O. Greep Award Lecture: Exploiting the Interface between Growth Factor [amp] Progesterone Receptor Signaling in Breast Cancer Models (8:00 AM - 9:15 AM) Roy O. Greep Award Lecture: Exploiting the Interface between Growth Factor [amp] Progesterone Receptor Signaling in Breast Cancer Models Carol A Lange University of Minnesota Cancer Research Center, Minneapolis, MN Nothing to Disclose: CAL 2012-06-25T08:00:00 Grand Ballroom AB 2012-06-25T00:00:00 1899-12-30T08:00:00 6124 248 1831 L5-1 PL05-1 Monday 253 2012


2977 ENDO12L_L5-2 PLENARY: TRANSLATIONAL - miR-33: Micro-Managing Lipoprotein Metabolism (8:00 AM - 9:15 AM) miR-33: Micro-Managing Lipoprotein Metabolism Kathryn J Moore New York University Medical Center, New York, NY Nothing to Disclose: KJM 2012-06-25T08:00:00 Grand Ballroom AB 2012-06-25T00:00:00 1899-12-30T08:00:00 6134 249 1832 L5-2 PL05-2 Monday 254 2012


2978 ENDO12L_CMF8-1 CASE MANAGEMENT FORUM: CLINICAL - Subtle/Subclinical Hyperthyroidism (8:30 AM - 9:15 AM) Subtle/Subclinical Hyperthyroidism Michael T McDermott University of Colorado Hospital, Denver, CO Nothing to Disclose: MTM 2012-06-25T08:30:00 Theater A 2012-06-25T00:00:00 1899-12-30T08:30:00 6177 250 1833 CMF8-1 CMF-TH1 Monday 255 2012


2979 ENDO12L_CMF8-2 CASE MANAGEMENT FORUM: CLINICAL - Subtle/Subclinical Hyperthyroidism (8:30 AM - 9:15 AM) Subtle/Subclinical Hyperthyroidism Hossein Gharib Mayo Clinic College of Medicine, Rochester, MN Nothing to Disclose: HG 2012-06-25T08:30:00 Theater A 2012-06-25T00:00:00 1899-12-30T08:30:00 6178 250 1834 CMF8-2 CMF-TH1 Monday 256 2012


2980 ENDO12L_M34 MEET-THE-PROFESSOR: CLINICAL - Making Sense of the Numbers: Diagnostic [amp] Treatment Thresholds of Diabetes during Pregnancy (8:30 AM - 9:15 AM) Making Sense of the Numbers: Diagnostic [amp] Treatment Thresholds of Diabetes during Pregnancy Susan Ellen Kirk University of Virginia, Charlottesville, VA Session supported by: Novo Nordisk Inc.[br][br]Nothing to Disclose: SEK 2012-06-25T08:30:00 Room 362 2012-06-25T00:00:00 1899-12-30T08:30:00 6180 252 1836 M34 D6 Monday 257 2012


2981 ENDO12L_M36 MEET-THE-PROFESSOR: CLINICAL - Clinical Management of Adrenocortical Carcinoma (8:30 AM - 9:15 AM) Clinical Management of Adrenocortical Carcinoma Martin Fassnacht University of Wuerzburg, Wuerzburg, Germany Nothing to Disclose: MF 2012-06-25T08:30:00 Room 320 2012-06-25T00:00:00 1899-12-30T08:30:00 6137 253 1837 M36 A2 Monday 258 2012


2982 ENDO12L_M38 MEET-THE-PROFESSOR: CLINICAL - Evaluation of Premature Adrenarche (8:30 AM - 9:15 AM) Evaluation of Premature Adrenarche Sharon E Oberfield Columbia University Medical Center, New York, NY Nothing to Disclose: SEO 2012-06-25T08:30:00 Room 310 2012-06-25T00:00:00 1899-12-30T08:30:00 6155 254 1838 M38 PED2 Monday 259 2012


2983 ENDO12L_M39 MEET-THE-PROFESSOR: CLINICAL - Pubertal Disruption in Chronic Inflammatory Disease (8:30 AM - 9:15 AM) Pubertal Disruption in Chronic Inflammatory Disease Mark Daniel DeBoer University of Virginia, Charlottesville, VA Disclosure Incomplete: MDD 2012-06-25T08:30:00 Room 360 2012-06-25T00:00:00 1899-12-30T08:30:00 6156 255 1839 M39 PED7 Monday 260 2012


2984 ENDO12L_M40 MEET-THE-PROFESSOR: CLINICAL - Specialized Lipid Testing: The Pre-Treatment Lipoprotein Size Consequences of Adiposopathy ([quot]Sick Fat[quot]) [amp] the Clinical Pitfalls of Post-Treatment Lipoprotein Particle Size Analyses (8:30 AM - 9:15 AM) Specialized Lipid Testing: The Pre-Treatment Lipoprotein Size Consequences of Adiposopathy ([quot]Sick Fat[quot]) [amp] the Clinical Pitfalls of Post-Treatment Lipoprotein Particle Size Analyses Harold Edward Bays L-MARC Research Center, Louisville, KY Disclosures: HEB: Advisory Group Member, Merck & Co.; Speaker, Merck & Co.; Researcher, Merck & Co., Orexigen, Vivus USA, Arena, Novo Nordisk. 2012-06-25T08:30:00 Room 371 2012-06-25T00:00:00 1899-12-30T08:30:00 6222 256 1840 M40 L1 Monday 261 2012


2985 ENDO12L_M41 MEET-THE-PROFESSOR: CLINICAL - What To Do If My Country Doesn[apos]t Have a FRAX? (8:30 AM - 9:15 AM) What To Do If My Country Doesn[apos]t Have a FRAX? Michael R McClung Oregon Osteoporosis Center, Portland, OR Nothing to Disclose: MRM 2012-06-25T08:30:00 Room 361 ABDE 2012-06-25T00:00:00 1899-12-30T08:30:00 6261 257 1841 M41 B11 Monday 262 2012


2986 ENDO12L_PS5-1 PRESIDENTIAL SYMPOSIUM SESSION: TRANSLATIONAL - Imaging [amp] Invasive Testing of the Pituitary, Parathyroid [amp] Adrenal Glands (9:30 AM - 11:00 AM) An Endocrinologist[apos]s View of Pituitary Imaging [amp] Pitfalls in Performing Inferior Petrosal Sinus Sampling Amir Hekmat Hamrahian Cleveland Clinic Foundation, Cleveland, OH Magnetic resonance (MR) imaging has become the imaging study of choice for evaluation of sellar and parasellar pathologies. In contrast, computerized tomography (CT) is better at evaluating bony changes and calcifications. The endocrinologist[apos]s review of pituitary images can be complementary to the radiologist[apos]s report and has the advantage of being able to take into account the clinical history and endocrine work up. Pituitary adenomas usually appear hypointense or isointense to the normal pituitary gland on noncontrast T1-weighted MR images; however, since they are less vascular than normal pituitary tissue, they usually appear hypodense postcontrast. Some radiological features, such as calcification on CT scan, a lack of sellar remodeling, intense enhancement postcontrast or a dural tail sign, suggest sellar lesions other than pituitary tumors. The cystic part of a sellar mass or a cystic lesion such as Rathke[apos]s cleft cyst appears hyperintense on T2- weighted MR images. Generalized pituitary gland enlargement may be seen during menopause, pregnancy or in patients with untreated primary hypothyroidism or hypogonadism, and its proper identification is important to prevent unnecessary surgery. Endocrinologists may be consulted about other sellar findings such as empty sella, sellar anatomical variations, or a prominent posterior pituitary that is mistaken for a sellar mass. Intrasellar aneurysms should always be considered in the differential diagnosis of sellar masses and further appropriate imaging studies performed when appropriate, to avoid a surgical catastrophe.[br]In patients with ACTH-dependent Cushing syndrome, Inferior petrosal sinus sampling (IPSS) is the gold standard test to differentiate between a pituitary and ectopic source but it does not reliably predict the lateralization of the pituitary adenoma. A number of pitfalls in assessing the IPSS result must be recognized. False-negative results (the presence of Cushing disease in the absence of a diagnostic IPS:P ACTH ratio) occur in 1-10% of patients. Measurement of prolactin during IPSS can reduce the false negative rate. An IPS:P prolactin ratio of [ge] 1.3 is suggestive of proper IPS venous sampling. A recent study suggests that the prolactin-adjusted ACTH ratio may be helpful for tumor localization in patients with Cushing disease, with a potential for a favourable impact on surgical outcomes if such a role is confirmed in larger studies.[br][br]Nothing to Disclose: AHH 2012-06-25T09:30:00 Grand Ballroom AB 2012-06-25T00:00:00 1899-12-30T09:30:00 2461 258 1842 PS5-1 SSS04 Monday 264 2012


2987 ENDO12L_PS5-2 PRESIDENTIAL SYMPOSIUM SESSION: TRANSLATIONAL - Imaging [amp] Invasive Testing of the Pituitary, Parathyroid [amp] Adrenal Glands (9:30 AM - 11:00 AM) Parathyroid Robert Udelsman Yale University School of Medicine, New Haven, CT Parathyroid imaging is a preoperative localization procedure to enhance operative planning and limit the extent of surgery. It is not a diagnostic procedure.[br]Imaging techniques vary in sensitivity, specificity, cost and exposure to ionizing radiation. Selection of one or more imaging techniques is dependent on the diagnosis of 1[deg] versus 2[deg] or 3[deg] hyperparathyroidism (HPTH), availability of minimally invasive surgical techniques and whether the patient has undergone previous exploration.[br]Parathyroid imaging is employed in the majority of patients with 1[deg] HPTH. Patients who require remedial exploration should always undergo preoperative imaging.[br]Noninvasive imaging include ultrasound, nuclear medicine studies, CT and MRI scans. These studies can be formatted to display simultaneous fusion images. All are dependent on the quality of the imaging device, technique and interpretation.[br]Parathyroid ultrasonography may demonstrate a hypoechoic ovoid lesion that is highly suggestive of an enlarged parathyroid gland. False positive lesions include lymph nodes and thyroid nodules. False negative studies are usually due to posterior or small parathyroid glands ([lt]100 mg) or those obscured by bony structures. Sestamibi scans employ early and delayed, duel isotope or single photon emission tomography imaging. The sensitivity and specificity of high quality ultrasound and sestamibi scans are similar at 60-80%.[br]CT and MRI scans are not routinely employed due to their moderate sensitivity (50-60%) and expense. Recently 4-dimensional CT scans have demonstrated increased sensitivity (60-80%) and can be particularly helpful in the remedial cases. Radiation exposure to the thyroid is not insignificant.[br]Invasive imaging include angiography with an arterial phase and venous access to sample parathyroid hormone (PTH). Invasive imaging is not indicated in [italic]de novo[/italic] cases. A less invasive technique is sonographically-guided PTH aspiration.[br]Parathyroid imaging is performed to help guide the surgeon especially when minimally invasive techniques are employed. Preoperative imaging is essential when remedial cervical exploration is required. Negative imaging is common and should not deter referral to an expert parathyroid surgeon because surgical cure rates far exceed the sensitivity of the imaging studies.[br][br]Nothing to Disclose: RU 2012-06-25T10:00:00 Grand Ballroom AB 2012-06-25T00:00:00 1899-12-30T10:00:00 2507 258 1843 PS5-2 SSS04 Monday 265 2012


2988 ENDO12L_PS5-3 PRESIDENTIAL SYMPOSIUM SESSION: TRANSLATIONAL - Imaging [amp] Invasive Testing of the Pituitary, Parathyroid [amp] Adrenal Glands (9:30 AM - 11:00 AM) Imaging and Invasive Testing of the Adrenal Glands: Focus on Primary Aldosteronism Michael Stowasser University of Queensland School of Medicine, Brisbane, Australia When an adrenal mass is detected by imaging, critical issues are whether it is benign or malignant (often difficult to determine by its imaging appearance) and whether it is autonomously producing hormone (including aldosterone [aldo], cortisol, sex steroids or catecholamines). In patients with primary aldosteronism (PA), imaging helps determine the subtype which is critical for guiding optimal treatment as unilateral PA is potentially curable by unilateral adrenalectomy whereas bilateral PA is usually treated medically. Adrenal CT misses [sim]50% of aldo-producing adenomas (APAs) and, like MRI, can be frankly misleading by demonstrating a non-functioning nodule in bilateral PA or contralateral to an APA. Se-cholesterol scanning misses most small tumors. Early experience with 11C-metomidate PET-CT scanning appears promising. CT is important for detecting large ([gt]2.5cm) adrenal lesions that may warrant removal based on malignant potential. It is also useful for localizing adrenal veins (AVs), facilitating successful cannulation during subsequent AV sampling (AVS). AVS is the only dependable way to differentiate unilateral from bilateral PA, and should be performed in all with PA who desire and are candidates for surgery. In our Centre, sampling is performed in the morning after overnight recumbency to avoid effects of diurnal variation and posture on steroid levels and stress is minimized. To improve chances of cannulation success, several samples are collected from each side. Our Centre accepts an AV/ peripheral venous (PV) cortisol gradient of [ge]3.0 to indicate successful cannulation. Although the right AV (RAV) is often harder to locate and cannulate than the left for anatomical reasons, in experienced hands RAV cannulation success rates approach [ge]90% and risk of adrenal haemorrhage is low. Rapid [ldquo]point-of-care[rdquo] plasma cortisol assays can be used to establish whether cannulation has been successful at the time of AVS, reducing the need for multiple samples and repeat procedures. Calculation of the aldo/cortisol ratio for each AV sample corrects for differences in [quot]dilution[quot] of AV with non-AV blood. In our Centre, if the aldo/cortisol ratio on one side is [gt]2 times higher, and on the other side no higher, than the simultaneous PV ratio, the study is considered to show lateralization. Reliable assays for aldo and cortisol are critical for correct AVS interpretation and new, rapid throughput mass spectrometric methods are an important advance.[br][br]Sources of Research Support: Irene Hunt Hypertension Research Trust.[br][br]Nothing to Disclose: MS 2012-06-25T10:30:00 Grand Ballroom AB 2012-06-25T00:00:00 1899-12-30T10:30:00 2485 258 1844 PS5-3 SSS04 Monday 266 2012


2989 ENDO12L_S35-1 SYMPOSIUM SESSION: TRANSLATIONAL - CNS Control of Islet Function (9:30 AM - 11:00 AM) Innervation Patterns of the Human Endocrine Pancreas Alejandro Caicedo University of Miami, Miami Disclosure Incomplete: AC 2012-06-25T09:30:00 Room 372 2012-06-25T00:00:00 1899-12-30T09:30:00 6103 259 1845 S35-1 T09-S05 Monday 268 2012


2990 ENDO12L_S35-2 SYMPOSIUM SESSION: TRANSLATIONAL - CNS Control of Islet Function (9:30 AM - 11:00 AM) CRF Regulation of Insulin Secretion Mark O Huising The Salk Institute for Biological Studies, La Jolla, CA The discovery in 1981 by Wylie Vale and colleagues of corticotropin-releasing factor (CRF) as the principal hypothalamic regulator of the hypothalamus-pituitary-adrenal axis represents a watershed moment in the field of neuroendocrinology. The decades that followed have come with the recognition that CRF signaling is key to many aspects of physiology and metabolism. Most recently, we identified the presence of a local CRF system including both types of CRF receptor within the endocrine pancreas. Stimulation of the CRF1 receptor on beta cells leads to an array of responses reminiscent of those induced by incretins. These include stimulation of intracellular cAMP and phosphorylation of CREB and Erk1/2, leading to immediate early gene expression and beta cell proliferation, and potentiation of glucose stimulated insulin secretion. The CRF2 receptor responds to the CRF-like peptide Urocortin 3, which is abundantly expressed in mature beta cells, and is required for full incretin- and glucose stimulated insulin secretion. Stress and metabolic challenges have marked effects on the pancreatic CRF system. Glucocorticoids (GCs) acutely and reciprocally affect the expression of both types of CRF receptor, desensitizing the islets to the insulinotropic actions of CRF-family peptides to facilitate the elevation in plasma glucose that is an integral part of [apos]fight or flight[apos]. While normal CRF receptor levels are restored within hours, chronic GC exposure associated with diseases such as Cushing[apos]s or the metabolic syndrome, or as a consequence of medication, could permanently offset the balance between expression of both types of CRF receptor in the islet and thus contribute to GC-induced forms of diabetes. Pancreatic expression of CRF receptors is also severely reduced in response to obesity, essentially abrogating their actions. Experimental interventions are unable to restore normal islet expression patterns of CRF receptors secondary to obesity-induced inhibition. The presence of CRFR1 and CRFR2 within the endocrine pancreas adds a novel layer of complexity to the intricate network of paracrine and autocrine factors and their cognate receptors whose coordinated efforts dictate islet hormone output and regulate functional beta cell mass. These findings instruct on how dysregulation of the intra-islet CRF system could contribute to the onset and progression of metabolic disease and identify novel targets in pursuit of strategies to counter its consequences.[br][br]Sources of Research Support: This work was supported by award P01DK026741-30 from the National Institute of Diabetes and Digestive and Kidney Diseases, the Clayton Medical Research Foundation, Inc., the Juvenile Diabetes Research Foundation, the Salk Center for Nutritional Genomics and the Leona M. & Harry B. Helmsley Charitable Trust.[br][br]Nothing to Disclose: MOH 2012-06-25T10:00:00 Room 372 2012-06-25T00:00:00 1899-12-30T10:00:00 2481 259 1846 S35-2 T09-S05 Monday 269 2012


2991 ENDO12L_S35-3 SYMPOSIUM SESSION: TRANSLATIONAL - CNS Control of Islet Function (9:30 AM - 11:00 AM) Hypothalamic Inflammation Disrupts Islet Function Licio Augusto Velloso State University of Campinas, Campinas, Brazil Nothing to Disclose: LAV 2012-06-25T10:30:00 Room 372 2012-06-25T00:00:00 1899-12-30T10:30:00 6104 259 1847 S35-3 T09-S05 Monday 270 2012


2992 ENDO12L_S36-1 SYMPOSIUM SESSION: BASIC - Coregulators of Nuclear Receptors (9:30 AM - 11:00 AM) Corepressors, Alternative mRNA Splicing, and Adipogenesis Martin Lawrence Privalsky University of California - Davis, Davis, CA The SMRT and N-CoR family of corepressors both regulate and help mediate the functions of a wide assortment of nuclear hormone receptors. Significantly, alternative mRNA splicing of NCoR and of SMRT creates a series of distinct corepressor [ldquo]variants[rdquo] that are expressed in different cell types, partner with different nuclear hormone receptors, respond to distinct kinase signals, and differ in their biological properties.[br]We will report our recent studies showing that different splice variants of the same corepressor can exert highly divergent functions. Most notably, we will describe how two distinct splice variants of NCoR regulate different target genes and play opposing roles in metabolic regulation and in adipocyte differentiation. [br]We will also describe how nuclear hormone receptor mutants associated with certain human endocrine and neoplastic diseases have acquired a novel preference for specific SMRT and N-CoR splice variants that may help explain previously confusing aspects of these disorders.[br]We conclude that NCoR and SMRT are each an extended family of interrelated, yet quite divergent transcriptional regulators, and that this diversity must be taken into account when interpreting the actions of these corepressors.[br][br]Sources of Research Support: RO1 DK53528, R37 CA53394.[br][br]Nothing to Disclose: MLP 2012-06-25T09:30:00 Room 320 2012-06-25T00:00:00 1899-12-30T09:30:00 450 260 1848 S36-1 T08-S05 Monday 272 2012


2993 ENDO12L_S36-2 SYMPOSIUM SESSION: BASIC - Coregulators of Nuclear Receptors (9:30 AM - 11:00 AM) Role of Coregulators in Antiestrogen and Antiandrogen Action Monica Marayag Montano Case Western Reserve University/School of Medicine, Cleveland, OH Hormone independence is a major problem in the treatment of hormone dependent cancers such as breast and prostate cancer, resulting in resistance to current cancer therapeutics. We have previously reported that Hexamethylene bis-acetamide Inducible Protein 1 (HEXIM1) inhibits Estrogen Receptor (ER) activity by competing with ER for binding to cyclin T1, a subunit of Positive Transcription Elongation b (P-TEFb). This results in the inhibition of the phosphorylation of RNA polymerase II (RNAPII) at serine 2 and the inhibition of transcription elongation of ER target genes. Since HEXIM1 can inhibit ER activity, we examined whether it plays a critical role in the inhibitory effects of tamoxifen on ER. We observed that tamoxifen induced HEXIM1 recruitment to the promoter region of ER target genes and decreased the recruitment of cyclin T1 and serine 2 phosphorylated RNAPII to the coding regions of these genes. Conversely, in cells wherein HEXIM1 expression has been downregulated we observed attenuation of the inhibitory effects of tamoxifen on estrogen-induced cyclin T1 recruitment to coding regions of ER target genes. As a consequence, downregulation of HEXIM1 resulted in the attenuation of the repressive effects of tamoxifen on estrogen-induced gene expression and proliferation. Conferring clinical relevance to our studies is our analysis of human breast cancer tissue samples that indicated association of lower expression of HEXIM1 with tumor recurrence in patients who received tamoxifen.[br]HEXIM1 also interacts with and inhibits transcriptional control by the Androgen Receptor (AR). Analyses of HEXIM1 expression using human tissue samples indicate decreased expression of HEXIM1 during the progression to castration resistant prostate cancer. As we observed in breast cancer cells, prostate cancer cells wherein HEXIM1 expression has been downregulated we observed attenuation of the inhibitory effects of bicalutamide on androgen-induced cyclin T1 and RNAPII recruitment to coding regions of AR target genes. Downregulation of HEXIM1 also resulted in the attenuation of the repressive effects of bicalutamide on androgen-induced gene expression and proliferation. Our studies provide a better understanding of the mechanistic basis for the inhibitory effect of antihormonal agents on nuclear hormone receptor activity and may suggest new therapeutic targets for the treatment of hormone resistant cancers.[br][br]Sources of Research Support: NIH CA92440 awarded to MMM.[br][br]Nothing to Disclose: MMM 2012-06-25T10:00:00 Room 320 2012-06-25T00:00:00 1899-12-30T10:00:00 2442 260 1849 S36-2 T08-S05 Monday 273 2012


2994 ENDO12L_S36-3 SYMPOSIUM SESSION: BASIC - Coregulators of Nuclear Receptors (9:30 AM - 11:00 AM) The p160 Steroid Receptor Coactivator (SRC) Family [amp] Breast Cancer Jianming Xu Baylor College of Medicine, Houston, TX The p160 SRC family contains three potent transcriptional coactivators for nuclear hormone receptors such as estrogen, progesterone and androgen receptors and certain other transcription factors such as E2F1, PEA3, AP-1 and HIF1. These p160 coactivators are expressed at low levels in normal mammary epithelial cells. However, the expression of SRC-1 and SRC-3 are significantly elevated in subsets of human breast cancers and their overexpressions are usually associated with disease recurrence, metastasis, endocrine therapy resistance and reduced disease-free survival. Using various genetically engineered breast cancer mouse models induced by oncogene expression or chemical carcinogen treatment and breast cancer cell line models, we found that knockout of SRC-1 specifically suppresses polyoma middle T (PyMT)-induced mammary tumor cell metastasis without affecting primary mammary tumor initiation and growth; knockout of SRC-2 has no effect on PyMT-induced mammary tumor development and metastasis; and knockout of SRC-3 represses mammary tumor initiation, growth and metastasis induced by oncogenes including PyMT and Ras and a chemical carcinogen. These results suggest that each SRC coactivator has a distinct role in breast cancer. Our laboratory also investigated the molecular mechanisms responsible for SRC-1 and SRC-3 to promote breast cancer growth and/or metastasis. We found that SRC-1 and SRC-3 serve as potent coactivators for multiple transcription factors including PEA3, AP-1 and HIF1 to directly upregulate Twist, integrin 5alpha, MMP2, MMP9, CSF-1 and/or VEGF, which in turn drives breast cancer cell epithelial mesenchymal transition (EMT), migration, invasion and metastasis. In addition, we also found that SRC-3 upregulates the expression of multiple components of the IGF-I signaling pathway to enhance breast cancer cell proliferation. This presentation will summarize these recent findings about the roles for the p160 SRC family members in breast cancer and discuss the related underlying molecular mechanisms. This presentation will also discuss the relevance and significance of these findings in the context of studies published by other laboratories.[br][br]Kuang S-Q et al., Cancer Res 2004; 64:1875. Kuang S-Q et al., Cancer Res 2005; 65:7993. Qin L et al., Mol Cell Biol 2008; 28:5937. Wang S et al., Proc. Natl. Acad. Sci. USA 2009; 106:151. Qin L et al., Cancer Res 2009; 69:3819. Qin L et al., Cancer Res 2011; 71:1742. Xu J et al., Nat Rev Cancer 2009; 9:615.[br][br]Sources of Research Support: NIH R01 CA112403, NIH R01 CA058242, and CPRIT RP101251-P4.[br][br]Nothing to Disclose: JX 2012-06-25T10:30:00 Room 320 2012-06-25T00:00:00 1899-12-30T10:30:00 2454 260 1850 S36-3 T08-S05 Monday 274 2012


2995 ENDO12L_S37-1 SYMPOSIUM SESSION: BASIC - Endocrine Regulation of Lipid [amp] Carbohydrate Metabolism, with Consequences for Longevity: Insights from Animal Models (9:30 AM - 11:00 AM) Nutrient Regulation of Lifespan in [italic]Drosophila[/italic] Scott Pletcher University of Michigan, Ann Arbor, MI Dietary restriction (DR; the limitation of nutrient availability without malnutrition) has effectively increased the lifespan of nearly every species to which it has been applied. In mammals, enhanced longevity is accompanied by a broad-spectrum improvement in health during aging. Work from a variety of laboratories using a range of simple model organisms has contributed to a model of DR that invokes the activation of specific neurons, including sensory neurons, and that involves neuroendocrine responses that target conserved pathways in peripheral tissues. Some of the target processes identified from this work, including respiration, xenobiotic detoxification, lipid metabolism, and protein homeostasis, have since been manipulated in mammalian systems to improve longevity or other health-related phenotypes. Indeed, translation of discoveries from simple models to more medically relevant systems has been surprisingly effective. Nevertheless, the development of successful human interventions requires a much better mechanistic understanding of how neuronal inputs are integrated and how critical metabolic processes imple-ment the changes that underlie the effects of DR. In this talk I will address several aspects of how nutrient modulate lifespan and healthspan in the fruit fly, [italic]Drosophila melanogaster[/italic]. I will discuss the impact of specific nutrients as well as the mechanisms by which sensory neurons of nutrients and the metabolism of specific amino acids affect longevity.[br][br]Sources of Research Support: The research was supported by US National Institutes of Health (R01AG030593 and R01AG023166), the Glenn Foundation, the American Federation for Aging Research, the Ellison Medical Foundation.[br][br]Nothing to Disclose: SP 2012-06-25T09:30:00 Room 342 ABDE 2012-06-25T00:00:00 1899-12-30T09:30:00 2436 261 1851 S37-1 T06-S01 Monday 276 2012


3245 ENDO12L1_7012 Hormone Health Network [ndash]Cooking with Pleasure (June 23-25 1:00 PM - 3:00 PM) Hormone Health Network [ndash] Cooking with Pleasure Supported by Boehringer Ingelheim Pharmaceuticals, Inc. [amp] Lilly USA, LLC, Genzyme Corporation, and Pfizer, Inc. 2012-06-23T13:00:00 George R. Brown Convention Center 2012-06-23T00:00:00 1899-12-30T13:00:00 7012 48 121 9910 Saturday 0 2012


3246 ENDO12L1_7013 CONFERENCE EVENT: Lipids and Obesity MOC Live Learning Session (6:00 PM - 9:00 PM) Lipids and Obesity MOC Live Learning Session Registration and fee required 2012-06-23T18:00:00 George R. Brown Convention Center 2012-06-23T00:00:00 1899-12-30T18:00:00 7013 116 918 9911 Saturday 0 2012


3247 ENDO12L1_7014 CMES ANCILLARY SYMPOSIUM: Insulin Therapy Is Still Hot and Getting Hotter [ndash] A Hot Seat Program (6:30 PM - 9:30 PM) Insulin Therapy Is Still Hot and Getting Hotter [ndash] A Hot Seat Program preregistration [br][br]Supported by Novo Nordisk, Inc. 2012-06-23T18:30:00 Hilton Lanier Ballroom (G- L) 2012-06-23T00:00:00 1899-12-30T18:30:00 7014 118 920 9912b Saturday 0 2012


3248 ENDO12L1_7015 CMES ANCILLARY SYMPOSIUM: Challenges in the Management of Overweight and Obese Patients: Targeting Weight Loss and Comorbidities (6:30 PM - 9:30 PM) Challenges in the Management of Overweight and Obese Patients: Targeting Weight Loss and Comorbidities preregistration [br][br]Supported by Eisai, Inc. 2012-06-23T18:30:00 Hilton Lanier Ballroom (A-F) 2012-06-23T00:00:00 1899-12-30T18:30:00 7015 117 919 9912a Saturday 0 2012


3249 ENDO12L1_7016 CONFERENCE EVENT: Women in Endocrinology Reception and Dinner (6:30 PM - 10:00 pm) Women in Endocrinology Reception and Dinner Registration and fee required 2012-06-23T18:30:00 Hilton Ballroom of the Americas (A,B,C) 2012-06-23T00:00:00 1899-12-30T18:30:00 7016 119 921 9913 Saturday 0 2012


3250 ENDO12L1_7017 CONFERENCE EVENT: ENDO Trivia Cup Challenge (6:45 PM - 8:30 PM) ENDO Trivia Cup Challenge 2012-06-23T18:45:00 George R. Brown Convention Center 2012-06-23T00:00:00 1899-12-30T18:45:00 7017 120 922 9914 Saturday 0 2012


3251 ENDO12L1_7018 CMES ANCILLARY SYMPOSIUM: Growth Hormone Deficiency Treatment in Pediatric and Adult GH Deficiency: Strategies, Efficacy, and Long-Term Safety (6:00 AM - 8:00 AM) Growth Hormone Deficiency Treatment in Pediatric and Adult GH Deficiency: Strategies, Efficacy, and Long-Term Safety preregistration [br][br]Supported by Novo Nordisk, Inc. 2012-06-24T18:00:00 Hilton Lanier Ballroom (A-F) 2012-06-24T00:00:00 1899-12-30T18:00:00 7018 246 1829 9915 Sunday 0 2012


3252 ENDO12L1_7019 CMES ANCILLARY SYMPOSIUM: Navigating the Data Stream: An Expert-Led Journal Club of GLP-1 Receptor Agonist Studies in Type 2 Diabetes (6:30 PM - 9:00 PM) Navigating the Data Stream: An Expert-Led Journal Club of GLP-1 Receptor Agonist Studies in Type 2 Diabetes preregistration [br][br]Supported by Novo Nordisk, Inc. 2012-06-24T18:30:00 Hilton Lanier Ballroom (G-L) 2012-06-24T00:00:00 1899-12-30T18:30:00 7019 247 1830 9916 Sunday 0 2012


3253 ENDO12L1_7020 CMES ANCILLARY SYMPOSIUM: Beyond Pituitary Surgery: Discussions on Cushing[apos]s Disease (6:00 AM - 8:00 AM) Beyond Pituitary Surgery: Discussions on Cushing[apos]s Disease preregistration [br][br]Supported by Corce 2012-06-25T18:00:00 Hilton Lanier Ballroom (A-F) 2012-06-25T00:00:00 1899-12-30T18:00:00 7020 352 2664 9917 Monday 0 2012


3254 ENDO12L1_7021 CMES ANCILLARY SYMPOSIUM: Hyperglycemia [amp] the Kidney: Emerging Therapies for Glycemic Control Therapy (6:30 PM - 9:00 PM) Hyperglycemia [amp] the Kidney: Emerging Therapies for Glycemic Control Therapy preregistration [br][br]Supported by Bristol-Myers Squibb and AstraZeneca LP 2012-06-25T18:30:00 Hilton Lanier Ballroom (A-F) 2012-06-25T00:00:00 1899-12-30T18:30:00 7021 353 2665 9918 Monday 0 2012