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Curcumin has well documented beneficial effects against diabetes but its poor bioavailability limits its use. Here, we investigated the effects of two monocarbonyl curcumin analogues - (2E,6E) -2,6-bis[ (2-trifluoromethyl) benzylidene]cyclohexanone (C66) and (2E,6E) -2,6-bis (2-bromobenzylidene) cyclohexanone) (B2BrBC) - on the expression of genes involved in insulin signaling and oxidative stress in pancreas samples from streptozotocin (STZ) -induced diabetic rats. Animals were divided into two experiments: (Experiment 1) STZ injection (60 mg/kg) followed by 30 days treatment with C66 or B2BrBC (both 50 µmol/kg, daily) or (Experiment 2) single-dose treated with C66 or B2BrBC (125 µmol/kg) , 6 hours before STZ injection, and sacrifice after 72 hours. Quantitative RT-PCR analysis was used to assess the expression of: insulin (Ins1) , insulin receptor substrate 2 (Irs2) , Slc2a2 solute carrier family 2 member 2 (Slc2a2) (s. Glut2) , mitogen-activated protein kinase 14 (Mapk14) , protein kinase AMP-activated catalytic subunit alpha 1 (Prkaa1) , peroxisome proliferator-activator receptor gamma (Pparg) , BCL2 apoptosis regulator (Bcl2) , BCL2-associated X apoptosis regulator (Bax) , and NF-κB inhibitor alpha (Nfkbia) . Results demonstrated that when C66 and B2BrBC were administered at lower, chronic doses after STZ application (Experiment 1) , they both reversed the effect of STZ on Irs2 mRNA levels in the pancreas, with the effect of C66 being significantly higher. At higher, acute doses (Experiment 2) , C66 ameliorated Ins1 mRNA levels after STZ treatment. Also, although not statistically significant, at higher doses, both C66 and B2BrBC showed a trend to normalize the suppressing effect of STZ on Glut2 mRNA levels. Overall, our data demonstrate that these two monocarbonyl curcumin analogues have direct effects on the expression of insulin signaling pathway-related genes in the pancreas. Disclosure N. Hadzi-Petrushev: None. M. Mladenov: None. J.B. Bogdanov: None. L. Poretsky: None. D. Avtanski: None. Funding Gerald J. and Dorothy R. Friedman New York Foundation for Medical Research

Objective: The diagnosis of diabetes mellitus is associated with an increased risk of hospital readmissions. The goal of this study was to determine whether there was a difference in the rates of 30-day and 365-day hospital readmissions between diabetic patients who, upon their discharge, received diabetes care in a standard primary care setting and those who received their care in a specialized multidisciplinary diabetes program. Methods: This was a randomized controlled prospective study. Results: One hundred and ninety two consecutive patients were recruited into the study, 95 (49%) into standard care (control group) and 97 (51%) into a multidisciplinary diabetes program (intervention group). The 30-day overall hospital readmission rates (including both emergency department and hospital readmissions) were 19% in the control group and 7% in the intervention group (P = .02). The 365-day overall hospital readmission rates were 38% in the control group and 14% in the intervention group (P = .0002). Conclusion: Patients with diabetes who are assigned to a specialized multidisciplinary diabetes program upon their discharge exhibit significantly reduced hospital readmission rates at 30 days and 365 days after discharge.

Polycystic ovary syndrome (PCOS) is a common endocrinopathy affecting 46XX individuals of reproductive age. Cardinal features of PCOS include hyperandrogenism, irregular periods, and insulin resistance. Pathogenesis is unclear but likely involves hypothalamic, pituitary, or ovarian abnormalities leading to increased androgen production. In addition, alternative insulin signaling pathways are activated to preserve ovarian sensitivity to insulin while other “classical” tissues (e.g. liver, adipose, muscle) are insulin resistant. Treatment targets specific symptoms and the most common regimens include weight loss, metformin, oral contraceptives, anti-androgen compounds, and fertility treatments. Observations of individuals with gene mutations affecting androgen metabolism suggest that androgens may influence the development of gender identity. We reviewed studies exploring the relationship between gender identity and PCOS to further elucidate this relationship. Rates of PCOS in hormone-naïve transmasculine (TM) individuals appear to be higher than in the general population as cited by small, early studies using convenience samples and inconsistent criteria for PCOS. A more recent, larger study using established guidelines for PCOS did not show this to be true. Further, other studies show that although PCOS patients are less likely to identify with a traditional feminine gender scheme compared to age-matched peers, the prevalence of gender incongruence in PCOS patients is not higher than in the general population. Larger systematic studies with control groups using modern diagnostic criteria for both PCOS and gender incongruence are needed to clarify the relationship between PCOS and gender identity.

Nonalcoholic Fatty Liver Disease (NAFLD) is a multifactorial metabolic disorder that was first described in 1980. It has been prevalent and on the rise for many years and is associated with other metabolic disorders such as obesity and type 2 diabetes mellitus (T2DM). NAFLD can be best described as a metabolic dysfunction that stems from insulin resistance-induced hepatic lipogenesis. This lipogenesis increases oxidative stress and hepatic inflammation and is often potentiated by genetic and gut microbiome dysfunction. As NAFLD progresses from simple steatosis to non-alcoholic steatohepatitis (NASH) and to cirrhosis and hepatocellular carcinoma (HCC), the odds of complications including cardiovascular disease (CVD), chronic kidney disease (CKD), and overall mortality increase. The aim of this review is to describe the metabolic causes and consequences of NAFLD while examining the risks that each stage of NAFLD poses. In this review, the etiology of “lean” NAFLD, the impact of obesity, T2DM, genetics, and microbiome dysbiosis on NAFLD progression are all explored. This review will also discuss the core issue behind the progression of NAFLD: insulin resistance (IR). Upon describing the causes and consequences of NAFLD, the effectiveness of diet modification, lifestyle changes, and glucagon-like peptide 1 receptor (GLP-1) agonists to retard NAFLD progression and stem the rate of complications is examined.

Aims/Hypothesis Hyperglycemia and hypoglycemia are associated with increased morbidity and mortality in the inpatient setting. Standard point of care capillary glucose testing (POCT) is commonly used in hospitalized patients to monitor their glucose levels. The goal of this study was to examine the relationships between the glucose readings obtained by a continuous glucose monitoring system (CGMS) (Freestyle Libre) and the capillary blood glucose results obtained by the inpatient glucose POCT meter (Accuchek Inform II) as well as between CGMS readings and the serum glucose values obtained by the hospital laboratory. Study participants had either primary or secondary diagnosis of diabetes mellitus and were admitted to non-critical units. We hypothesized that there exists an acceptable agreement between the capillary blood glucose results obtained by the inpatient glucose POCT meter (Accuchek Inform II) and the readings obtained by the CGMS (Freestyle Libre); and that there exists an acceptable agreement between the serum glucose levels and the glucose values obtained by the CGMS. Methods This was an Institutional Review Board approved prospective cohort study for the non– critical inpatient setting. Fifty-two hospitalized patients with diabetes were recruited. After informed consent was obtained, patients were instructed on the application and use of the CGMS. The data were assessed using a standard regression analysis and modified Bland Altman analysis. All analyses were conducted using SAS, release 3.8 Enterprise Edition (SAS Institute Inc., Cary, NC). Results Fifty-two subjects recruited into the study represented a sample of convenience. There were a total of 467 AccuChek-Libre pairs, The regression analysis showed a negative bias between. Libre and AccuChek, R2 = 0.83, with Libre glucose readings on average being lower than those of AccuChek. Using Bland-Altman analysis, 42% of the 467 Libre-AccuChek pairs had a difference in glucose reading more than 15%. Mean absolute relative difference (MARD) between Libre and AccuChek was 15.6%; mean relative difference (MRD) between Libre and AccuChek was −11.4%. The regression analysis showed a negative bias between Libre and serum glucose, R2 = 0.89. Using Bland Altman analysis, 36% of the 44 Libre-serum pairs had a difference in glucose reading more than 15%. Mean absolute relative difference (MARD) between Libre and serum glucose was 13.2%; mean relative difference (MRD) between Libre and serum glucose was −12.5%. A review of the data pairs showed that 71/467 Accuchek-Libre pairs had one result that was either below 70 mg/dl or above 200 mg/dl (combined American Diabetes Association-ADA-, American College of Physicians-ACP- and American College of Endocrinology-AACE- goals). Thus 85%, of these pairs would have yielded results that engendered the same intervention (e.g. treatment for hypoglycemia or hyperglycemia). Likewise 5/45 Serum-Libre pairs had one result that was either below 70 mg/dl or above 200 mg/dl; thus 89% of these pairs would have yielded results requiring the same intervention. Conclusion/Interpretation These findings confirm the existent literature and indicate acceptable agreement between the standard POCT and the CGMS as well as between serum glucose and the CGMS values. Because of the advantages of the CGMS over capillary blood glucose testing (reduced patient discomfort and reduced staff exposure to patients in isolation) CGMS use may be preferable to the current bedside capillary blood glucose testing in hospitalized patients with diabetes mellitus. As with other laboratory measures, clinical judgement needs to be exercised when the laboratory values are used to guide patient care.

Background Animal models of diet‐induced obesity (DIO) are commonly used in medical research for mimicking human diseases. There is no universal animal model, and careful evaluation of variety of factors needs to be considered when designing new experiments. Here, we investigated the effect of 9 weeks high‐fat diet (HFD) intervention, providing 60% energy from fat, on parameters of inflammation and insulin resistance in male C57BL/6J mice. Methods Six weeks old mice were initiated on regular diet (RD) or HFD providing 60 kcal energy from fat for 9 weeks. Fasting blood glucose levels were measured by glucometer, and fasting plasma levels of insulin and proinflammatory cytokines by Luminex assay. Insulin sensitivity was evaluated by using QUICKI and HOMA2 indexes. Results HFD mice showed ~ 40% higher body weight and ~ 20% larger abdominal circumference, due to an increase in the white adipose tissue mass. Liver examination revealed increased size and higher hepatic lipid accumulation in livers from HFD mice compared to their RD counterparts. Animals from the HFD group were characterized with significantly higher presence of crown‐like structures (CLS) in WAT and higher plasma levels of proinflammatory cytokines (TNF‐α, IL‐6, leptin, MCP‐1, PAI‐1, and resistin). HFD‐fed mice also demonstrated impaired insulin sensitivity (lower QUICKI, higher HOMA‐insulin resistance (HOMA‐IR), and lower HOMA‐percent sensitivity (HOMA‐%S)) index values. Conclusion Male C57BL/6J mice on 9 weeks HFD providing 60 kcal energy from fat display impaired insulin sensitivity and chronic inflammation, thus making this DIO mouse model appropriate for studies of early stages of obesity‐related pathology.

Energy metabolism and reproduction are closely linked; however, the role of the individual energy metabolism hormones in this association is still poorly understood. Irisin is a hormone produced mostly by skeletal muscle cells during exercise or shivering, and leptin is a satiety adipokine synthesized by the white adipose tissue. The aim of this study was to investigate the in vitro effects of irisin and leptin on steroidogenesis in cultured human granulosa cells. The cells were purified from follicular fluid samples from patients undergoing in vitro fertilization (IVF). Primary cell cultures were treated with increasing concentrations of irisin (0, 125, 250, 500, 1,000, and 2,000 ng/ml) or leptin (0, 25, 50, 100, 200, and 400 ng/ml) for 24 hours and subjected to qRT-PCR analysis for the expression of key steroidogenic enzymes (CYP11A1, CYP19A1, CYP21A1, HSD3B1, and HSD17B3). Concentrations of progesterone, testosterone and estradiol in the conditioned cell culture medium were measured using ELISA. Results demonstrated that both irisin and leptin significantly upregulated CYP19A1 mRNA expression. Leptin also increased the expression of HSD3B1 mRNA levels. Although not statistically significant, leptin produced a trend of increasing estradiol production after 24 hours of treatment. Taken together, these data support the hypothesis that irisin and leptin may affect steroid hormone metabolism in the ovary by modulating the expression of key steroidogenic enzymes. Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

Introduction: Standard point of care capillary blood glucose testing (POCT) is commonly used in hospitalized patients to monitor their glucose levels. This technique is associated with increased staff/patient contact and patient discomfort. We examined the correlation of the capillary blood glucose results obtained by the POCT device (Accuchek Inform II) with the serum glucose values obtained by the hospital laboratory and with the glucose readings obtained by the FDA-waived for in-patient use (because of COVID-19 pandemic) Freestyle Libre continuous glucose monitoring system (CGMS). Methods: For this prospective cohort study fifty-two adult participants who were diagnosed with diabetes mellitus and admitted to non-critical units were recruited. Among them, 41 (79%) were male, 50 (96%) had type 2 diabetes mellitus. CGMS sensors were placed on patients’ arms, POCT and serum glucose measurements were taken as ordered. Descriptive statistics were calculated and linear mixed models were used to determine correlation. Results: There were a total of 467 Accuchek-Libre pairs with a highly statistically significant association between pairs, p Conclusion: These findings indicate excellent correlation between the standard POCT and the CGMS as well as between serum glucose and the CGMS values. Because of the advantages of the CGMS glucose readings over blood capillary glucose testing (reduced patient discomfort and reduced staff exposure to patients during the pandemic) hospitals should consider replacing capillary blood glucose testing with CGMS. Disclosure R. Murray-bachmann: None. K. Ziskovich: None. M. Sarbanes: None. T. Leung: None. A. Myers: None. S. Murthi: None. L. Poretsky: None.

Background The association between coronary artery disease (CAD) and diabetes mellitus (DM) is strong but the physiologic mechanisms responsible for this association remain unclear. Patients with DM exhibit high circulating levels of glycated proteins and lipoproteins called advanced glycation end products (AGEs) which have been implicated in the development of oxidative damage to vascular endothelium. We examined the relationships between the presence and extent of CAD and AGEs in patients undergoing elective coronary artery catheterization in an urban teaching hospital. Methods Patients with possible CAD (n = 364) were recruited prior to elective cardiac catheterization (52% male, 48% diabetic). Regression and correlation analyses were used to examine the relationship between serum AGE concentrations, soluble AGE receptor (sRAGE) concentration, HbA1c, LDL and the presence of obstructive CAD along with the burden of CAD measured by SYNTAX and SYNTAX II scores. Results AGE and sRAGE levels did not significantly correlate with any of the studied coronary artery disease parameters. HbA1c showed positive correlation with both SYNTAX and SYNTAX II scores in patients with and without diabetes. Conclusion In this cross-sectional study of patients with possible CAD, serum AGEs and sRAGE concentrations did not correlate with SYNTAX or SYNTAX II scores regardless of diabetic status. HbA1C correlated positively with the SYNTAX and SYNTAX II scores in both diabetic and non-diabetic populations., Highlights • Serum AGE levels do not correlate with severity of CAD as measured by SYNTAX or SYNTAX II scores. • SYNTAX scores are correlated with HbA1C regardless of diabetes status. • Serum AGE levels do not correlate with LDL levels.

Introduction: DM is associated with a higher risk of readmissions. We examined the effects of a structured diabetes outpatient program on RR. Methods: Patients hospitalized with a primary or secondary diagnosis of DM were randomized to standard care (control group [C]) or a structured diabetes program (intervention group [I]) upon discharge. The structured program included a visit to the CDE nurse practitioner (NP) who developed individualized plan for every subject. Subsequent visits were with an NP, nutritionist, social worker and endocrinologist as needed. RR data were generated from electronic medical records and included emergency department (ED) as well as hospital admissions. RR were calculated by dividing the number of readmissions by the number of initial (index) admissions and expressed as %. Results: 192 subjects were recruited (95 into [C] and 97 into [I]). The mean age was 56±13 yrs [C] and 60±12 yrs [I]. The [C] had 56 (29%) males and 39 (20%) females; [I] had 60 (31%) males and 37 (19%) females. 66 (35%) subjects were African American, 2 (1%) American Indian, 1 (0.5%) Hawaiian, 57 (30%) Caucasian, 11 (5%) Asian, 12 (6%) identified as more than one race, 43 (22%) of unknown race. There were no statistically significant differences between the groups in these characteristics. The 30-day RR: overall (ED and Hospital): 19% [C] and 7% [I], p=0.01; Hospital: 9% [C] and 4% [I], p=0.14; ED: 17% [C] and 4% [I], p=0.003. The 365-day RR: overall 38% [C] and 14% [I], p=0.0002; Hospital: 33% [C] and 8% [I], p Conclusion: A structured outpatient diabetes program reduced RR in DM patients when compared with standard care. 30 day RR were reduced primarily due to a reduction in the ED visits, whereas 365 day RR were reduced due the reduction in both ED visits and hospital admissions. Larger multicenter studies are needed to confirm these data. Disclosure A. Bhalodkar: None. H. Sonmez: None. M. Lesser: None. T. Leung: None. K. Ziskovich: None. D. Inlall: None. L. Poretsky: None. M. Krymskaya: None. R. Murray-Bachmann: None.

Hypoxia is a common phenomenon in solid tumor development caused by a decrease in either oxygen concentration or oxygen pressure as a result of rapid tumor cell growth. Hypoxia is characterized by stabilization of the alpha subunit of the hypoxia-inducible factor (HIF-1α) and its nuclear translocation and heterodimerization with HIF-1β. Activation of this signaling pathway involves multiple downstream effectors including carbonic anhydrase 9 (CA9, s. CAIX). A reliable method to mimic hypoxia utilizes cobalt(II) chloride (CoCl2), which directly induces the expression of HIF-1α. The aim of this study was to optimize the experimental conditions for CoCl2 treatment of breast cancer cells in vitro using three human breast cancer cell lines (MDA-MB-231, T-47D, and MCF-7 cells). We performed time- and concentration-response experiments, using various concentrations of CoCl2 (50, 100, 200, and 300 μM) for 24 and 48 hours, and measured the expression of HIF-1α and CA9 by qRT-PCR and Western blot analyses. Results demonstrated that CoCl2 downregulated HIF-1α mRNA levels but upregulated CA9 mRNA levels in a concentration- and time-dependent manner. Concomitantly, CoCl2 treatment resulted in a significant induction of HIF-1α protein levels. We further investigated the effect of the CoCl2 concentrations listed above on cell apoptosis using an in situ apoptosis detection kit. The results demonstrated that concentrations of CoCl2 up to 100 μM had no significant effect on cell apoptosis.

Inflammation is a critical component of tumor initiation and progression. Chronic inflammation triggers molecular events that can promote carcinogenesis, tumor vascularization and metastasis. As inflammatory mediators, cytokines play an important role in the interplay between the tumor cells and tumor microenvironment. Cytokines released by the tumor-associated macrophages modulate cancer cell survival, stemness, invasiveness, and tumor vascularization. Breast cancer cells, however, also produce a variety of cytokines, whose role in cancer development is poorly understood. The aim of our study was to characterize the basal cytokine secretory activity in commonly used human breast cancer cell lines (MDA-MB-231, MCF-7, BT-474, and T-47D). Using MILLIPLEX assay, we measured the expression of 41 cytokines, including interleukins, monokines, interferons and growth factors. We also compared cytokine expression profile of breast cancer cells with those of non-tumorigenic human breast epithelial MCF-10A cells. Further, we investigated whether hypoxia modulates cytokine secretion of breast cancer cells. Using cobalt(II) chloride (CoCl2) to mimic hypoxia, we compared the effect of various treatment doses and intervals on cytokine production in the breast cancer cells. Results demonstrated that CoCl2 affects the release of multiple cytokines in a dose- and concentration-dependent manner, thus highlighting the role of cancer cell-derived cytokines on breast tumor progression. Understanding the molecular actions of cytokines in the tumor microenvironment is important for understanding the mechanism of cancer initiation and progression.

Fertility and energy metabolism are closely associated, and the cytokines produced by the adipose and muscle tissue play a role in this association. Leptin, predominantly produced by the white adipose tissue, and irisin, produced by the brown adipose and skeletal muscle tissues, are cytokines that are important in balancing energy metabolism. This study aimed to investigate the effects of leptin and irisin on steroidogenic enzyme gene expression in human ovarian granulosa cells in vitro. Granulosa cells were retrieved and isolated from ovarian follicular fluid during in vitro fertilization (IVF) procedures. Cells were placed in primary in vitro cultures and treated with increasing concentrations of leptin (25, 50, 100, 200, and 400 ng/ml) or irisin (125, 250, 500, 1,000, and 2,000 ng/ml) for 24, 48, and 72 hours. mRNA expression levels of CYP11A1, CYP19A1, CYP21A2, HSD3B1, and HSD17B3 were measured by qRT-PCR analysis. Leptin treatment of granulosa cells resulted in significant upregulation of CYP21A2 mRNA levels, while irisin significantly downregulated mRNA levels of CYP11A1, CYP19A1, and HSD3B1. Taken together, these early experiments demonstrate that leptin and irisin may affect steroid hormone production in the ovary by targeting the gene expression of key steroidogenic enzymes. Additional experiments are in progress.

Metastatic breast cancer is the most advanced stage of breast cancer and the leading cause of breast cancer mortality. Although understanding of the cancer progression and metastasis process has improved, the bi-directional communication between the tumor cell and the tumor microenvironment is still not well understood. Breast cancer cells are highly secretory, and their secretory activity is modulated by a variety of inflammatory stimuli present in the tumor microenvironment. Here, we characterized the cytokine expression in human breast cancer cells (MDA-MB-231, MCF-7, T-47D, and BT-474) in vitro using 41 cytokine MILLIPLEX assay. Further, we compared cytokine expression in breast cancer cells to those in non-tumorigenic human breast epithelial MCF-10A cells.

Advanced glycation end-products (AGEs) are endogenously produced or exogenously derived glycated proteins and lipoproteins. The accumulation of AGEs may have a variety of adverse effects since they cause oxidative stress and thus may act as contributing factors to the development of disease states such as, for example, complications of diabetes mellitus. The aim of this study was to investigate whether the levels of three AGEs (pentosidine, N(6)-carboxymethyllysine [CML], and methylglyoxal [MGO]) and the soluble form of the AGE receptor (sRAGE) may be used as markers for the presence and extent of coronary artery disease (CAD). We recruited 363 consecutive patients with suspected CAD who presented for cardiac catheterization at Lenox Hill Hospital in New York City (54% of patients were male, 48% of the patients had diabetes). At the time of this abstract submission 169 patients had circulating pentosidine levels measured (74 of these patients had diabetes). We examined relations between pentosidine levels, HbA1c, LDL, and the extend of CAD (measured by SYNTAX score). Serum pentosidine levels did not significantly correlate with any of other parameters. There was, however, a positive correlation between HbA1c levels and the SYNTAX score (Spearman correlation coefficients of 0.26, ρ = 0.0001 for the entire group; 0.18, ρ = 0.01 for patients without diabetes; and 0.29, ρ= 0.0001 for patients with diabetes). In conclusion, our preliminary results demonstrate that in patients with suspected CAD undergoing cardiac catheterization, SYNTAX score does not correlate with pentosidine levels but correlates positively with HbA1c. The data on all three AGEs and sRAGE in the entire cohort of 363 patients will be presented at the meeting. Disclosure C. Basman: None. D. Avtanski: None. K. Ziskovich: None. R. Jonas: None. S. Fishman: None. H. Sonmez: None. U. Rashid: None. A. Kodra: None. G. Stoffels: None. M. Lesser: None. V. Singh: None. L. Poretsky: None. Funding The Gerald J. and Dorothy R. Friedman New York Foundation for Medical Research

Resistin is an adipokine produced by the white adipocytes and adipose-derived macrophages, which mediates inflammation and insulin resistance Huang et al., 1997 and Renehan et al., 2008 Feb. Here, we provide data on the effect of resistin on epithelial to mesenchymal transition (EMT) in breast cancer cells in vitro. As model systems, we used human MCF-7 (low-metastatic) and MDA-MB-231 (high-metastatic) breast cancer cell lines. To optimize experimental conditions, we treated the cells with various concentrations of resistin (12.5, 25 and 50 ng/ml) for different time intervals (6 and 24 hours), and measured SOCS3 mRNA expression by using qRT-PCR analysis. Further, we used qRT-PCR and Western blot analyses to measure the expression of various epithelial (E-cadherin, claudin-1) and mesenchymal (SNAIL, SLUG, ZEB1, TWIST1, fibronectin, and vimentin) markers after resistin treatment. This data article is part of a study Avtanski et al., 2019 May, where detailed interpretation and discussion can be found. Keywords: Resistin, Epithelial to mesenchymal transition (EMT), Breast cancer

Resistin is an adipokine produced by the white adipose tissue that associates with breast cancer progression. Our previous studies demonstrated that resistin increases cancer motility by inducing epithelial to mesenchymal transition (EMT) and acquisition of cancer stem cell properties in breast cancer cells. AXL is a tyrosine kinase receptor belonging to the tumor-associated macrophage family which regulates a variety of processes including cell survival, growth, aggregation, migration, etc. Previous reports indicate that AXL also plays a significant role in tumor growth and progression, suppressing cell apoptosis and inducing angiogenesis and cancer metastasis. The aim of this study was to investigate whether the resistin effects on EMT are mediated by AXL. Using quantitative PCR array followed by qRT-PCR analysis, we found that in MCF-7 and MDA-MB-231 breast cancer cells resistin significantly upregulated mRNA expression levels of AXL and mesenchymal cell markers (SNAIL, SLUG, and ZEB1 transcription factors and their target genes fibronectin and vimentin), concomitantly inducing SNAIL and ZEB1 nuclear translocation. In order to investigate whether AXL participates in the molecular mechanism by which resistin affects EMT, we used short-interfering RNA to silence AXL mRNA expression or a chemical approach (R428) to suppress AXL activity. Either, AXL siRNA or R428, significantly blocked the effects of resistin on the expression of mesenchymal markers as well as inhibited resistin-induced nuclear translocation of SNAIL and ZEB1. Taken together, these results demonstrate that activation of AXL mediates the effects of resistin on EMT. Additional experiments are in progress.

Resistin is a proiflammatory adipokine produced by the white adipose tissue macrophages and adipocytes. Resistin expression is upregulated in breast cancer tissue and positively correlates with tumor stage, tumor size and lymph node metastasis. Previous studies found that resistin increases breast cancer invasiveness, stimulates epithelial to mesenchymal transition (EMT) and induces acquisition of cancer stem cell properties in breast cancer cells. Breast cancer cells express a variety of cytokines and their receptors, and resistin mRNA expression was previously detected in HCT-116 colorectal cancer cells. By using qRT-PCR analysis, we amplified resistin mRNA and compared its relative expression levels in a panel of breast cancer and breast non-carcinogenic cell lines, including MCF-7, T47D, BT474, HCC1806, MDA-MB-468, MDA-MB-231, and MCF-10A cells. We found that MCF-7, T47D, and HCC1806 cells express detectable levels of resistin mRNA. Using MCF-7 cells, we further investigated whether this expression can be modulated by treatment with different cytokines (resistin, TNFα, or TGFβ). Our results indicated that resistin (in concentration of 12.5 ng/ml), TNFα and TGFβ (in concentrations of 10 ng/ml) upregulated the expression of resistin mRNA in a time-dependent manner. In conclusion, we report that breast cancer cells express resistin mRNA, and that this expression is upregulated after treatment with resistin, TNFα, and TGFβ. To our best knowledge, this is the first study describing resistin mRNA expression in breast cancer cells and its alteration by proinflammatory factors. Further experiments are in progress.

Transgender individuals are medically underserved in the United States. Based on the National Transgender Discrimination Survey, 19% transgender population were refused healthcare, 50% had providers with no knowledge of transgender care and 28% had to delay medical care because of discrimination based on the sexual orientation [1]. The Friedman Transgender Health and Wellness Program (FTHWP) was established in 2017 at Lenox Hill Hospital to provide clinical, legal and social services to transgender individuals. These services include pediatric and adult endocrinology services, primary care, mental health, gynecological services, HIV/AIDS testing and care, referrals for hair removal, vocal cord training and gender confirming surgery, legal services, specialty referrals and social work assistance. There were 10 participants at FTHWP in June 2017. The number of participants increased to 60 by September 2018. The patients’ ages range from 11 years to 80 years old. 32 are Caucasians (53%), 13 are African American (21%), 10 are Asian (16%) and 6 are Hispanic (10%). 29 patients are trans-male, 28 patients are trans-female, 4 patients are gender-fluid. 39 patients have received previous hormone therapy, while 22 patients initiated hormonal therapy in our program. From this limited initial experience we concluded that the demand for comprehensive transgender medical services exists and that the patients display diverse demographic and clinical characteristics. Endocrinologists are well positioned to establish transgender health centers because of their background in both internal medicine and hormone therapy. Reference: [1] Grant JM, Mottet LA, Tanis J, Harrison J, Herman JL, Keisling M. Injustice at Every Turn: A Report of the National Transgender Discrimination Survey. Washington, DC: National Center for Transgender Equality and National Gay and Lesbian Task Force; 2011.

Breast cancer incidence and metastasis in postmenopausal women are known to associate with obesity, but the molecular mechanisms behind this association are largely unknown. We investigated the effect of adipokine resistin on epithelial to mesenchymal transition (EMT) and stemness in breast cancer cells in vitro. Previous reports demonstrated that the inflammatory actions of resistin are mediated by the adenylyl cyclase-associated protein 1 (CAP1), which serves as its receptor. As a model for our study, we used MCF-7 and MDA-MB-231 breast cancer and MCF-10A breast epithelial cells. We showed that in MCF-7 cells resistin increases the migration of MCF-7 and MDA-MB-231 cells and induces the formation of cellular protrusions through reorganization of F-actin filaments. Resistin upregulated the expression of mesenchymal markers involved in EMT (SNAIL, SLUG, ZEB1, TWIST1, fibronectin, and vimentin), and downregulated those of epithelial markers (E-cadherin and claudin-1). Resistin also potentiated the nuclear translocation of SNAIL protein, indicating initiation of EMT reprogramming. We further induced EMT in non-carcinogenic breast epithelial MCF-10A cells demonstrating that the effects of resistin on EMT were not breast cancer cell specific. In order to assess whether resistin-induced EMT depends on CAP1, we used siRNA approach to silence CAP1 gene in MCF-7 cells. Results demonstrated that when CAP1 was silenced, the induction of SNAIL, ZEB1 and vimentin expression by resistin as well as SNAIL and ZEB1 nuclear translocation, were abolished. Additionally, CAP1 silencing resulted in a suppression of MCF-7 cells migration. We performed quantitative PCR array profiling the expression of 84 genes related to cancer stem cells (CSC), pluripotency and metastasis and selected a set of genes (ALDH1A1, ITGA4, LIN28B, SMO, KLF17, PTPRC, PROM1, SIRT1, and PECAM1) that were modulated by resistin. Further experiments demonstrated that the effect of resistin on the expression of some of these genes (PROM1, PTPRC, KLF17, SIRT1, and PECAM1) was also dependent on CAP1. Our results demonstrate that resistin promotes the metastatic potential of breast cancer cells by inducing EMT and stemness and some of these effects are mediated by CAP1.

Individuals who identify as transgender are often faced with social, economic, medical, and political barriers that require guidance from knowledgeable organizations. Such organizations have experience in providing medical care, counseling, support groups, legal assistance, and social services. We have therefore compiled a list of some of the notable national and international resources for transgender individuals. Here we describe some of these in more detail.

Since the mid-twentieth century, transgender individuals have become increasingly visible. Strong advocacy group efforts and increasing government support have improved access to medical care for people with gender dysphoria. Physicians should be aware of the unique conditions and challenges affecting this population. Healthcare professional organizations such as the American Medical Association (AMA), the Endocrine Society, and the World Professional Association for Transgender Health have concluded that hormonal and surgical treatment of gender dysphoria is medically necessary to prevent long-term morbidity. Furthermore, physicians caring for transgender persons must have sufficient experience to recognize gender dysphoria as a spectrum of conditions, and should be adept in tailoring therapy to the individual patient. Many, but not all, gender dysphoric individuals presenting for care will ultimately seek endocrine therapy for the modulation of endogenous hormone production and exogenous hormone supplementation, to improve their quality of life.

Resistin is a pro-inflammatory adipokine produced by the white adipose tissue (WAT) adipocytes and macrophages. Obesity results in chronic inflammation of the WAT, marked by an increase in resistin and other inflammatory cytokines, and by infiltrating leukocytes. Elevated resistin levels are believed to play a major role in the development of insulin resistance in the peripheral tissues. Adenylyl cyclase-associated protein 1 (CAP1) was recently identified as a receptor for resistin. In the present study we aimed to investigate whether CAP1 mediates resistin actions which may affect insulin sensitivity in the liver. As a model we used BNL CL.2 mouse liver cell line. Concentration- and time-dependent experiments demonstrated that resistin upregulated TNFα, SOCS3, IL-1α, and IL-6 mRNA expression maximally when used in concentration of 12.5 ng/ml for 6 hours. In order to determine the CAP1 involvement in mediating resistin actions in the liver, we transfected BNL CL.2 cells with CAP1 siRNA and performed a real-time PCR array measuring the expression of 84 key genes involved in insulin signaling, adipokine signaling, and inflammation. Results demonstrated that resistin upregulated mRNA expression of IL-6; this effect was ameliorated when CAP1 was downregulated. Knock-down of CAP1 facilitated mRNA expression of genes involved in insulin signaling and adipokine signaling pathways, while it resulted in downregulation of infiltrating leukocyte markers expression. Taken together these results indicate that CAP1 is a mediator of resistin actions in the liver. Disclosure D. Avtanski: None. A. Garcia: None. P. Thangeswaran: None. B. Caraballo Bordon: None. L. Poretsky: None.

Polycystic ovary syndrome (PCOS), a heterogeneous and chronic condition, today affects about 5% of women of reproductive age. PCOS is strongly associated with states of insulin resistance and hyperinsulinemia. Risk factors include genetics, metabolic profiles, and the in utero environment. Long-term consequences of PCOS include metabolic complications such as diabetes, obesity, and cardiovascular disease. Dysregulation of insulin action is closely linked to the pathogenesis of PCOS. However, whether insulin resistance is the causative factor in the development of PCOS remains to be ascertained. Moreover, the mechanism by which insulin resistance may lead to reproductive dysfunction requires further elucidation.

The insulin/insulin-like growth factor (IGF) pathways and glucose metabolism act as mediators of human ovarian function and female fertility. In normal insulin action, insulin binds to its own receptors in the ovary to mediate steroidogenesis and act as a co-gonadotropin. Insulin with other factors may influence ovarian growth and cyst formation. The IGF pathway also seems to influence normal ovarian function. Insulin signaling affects reproductive function. Dysregulation of this pathway leads to altered puberty, ovulation, and fertility. Better understanding of the normal physiology and pathophysiology of insulin, IGF, and glucose effects on the human reproductive system will allow for better outcomes.

The recently discovered myo- and adipokine irisin affects insulin sensitivity in classical insulin target tissues (adipose tissue, skeletal muscle and liver), but the reproductive effects of this hormone, if any, remain largely unexplored. We hypothesized that irisin may have effects on the hypothalamic-pituitary-gonadal axis. To test this hypothesis, we used murine pituitary mPit12 and human ovarian granulosa cells. GnRH treatment resulted in significant (up to 2.5-fold, p