bluuebunny/crossref_metadata_2025_split huggingface dataset:
OR02-05 Effect of Testosterone Replacement Therapy Added to Intensive Lifestyle Intervention on Cognitive Functions in Frail, Older Veterans with Hypogonadism and Obesity: A Randomized Clinical Trial
Authors: Gregori Giulia , Celli Alessandra , Barnouin Yoann , Paudyal Arjun , Villareal Reina Condevillamar , Quallas Clifford , Villareal Dennis Tan 
Published: May 2020

Abstract
Background: Both hypogonadism and obesity are common in older men which might additively exacerbate their age-related decline in cognitive functions. We tested the hypothesis that the addition of testosterone replacement therapy to intensive lifestyle intervention would enhance the benefits of intensive lifestyle intervention on cognition in older men with hypogonadism and obesity.
Methods: Eighty-three older (age≥65 years) male veterans with obesity (BMI≥30 kg/m2) and evidence of persistently low AM testosterone (&lt;300 ng/dl) associated with frailty (modified Physical Performance Test score &lt;31) were randomized to six months of: 1) lifestyle therapy (diet-induced weight loss and supervised aerobic and resistance exercise training) + testosterone replacement therapy (LT+Test) or 2) lifestyle therapy + placebo (LT+Pbo). In this secondary analyses, outcomes were changes in cognition as assessed through comprehensive cognitive test battery (Modified Mini-Mental State Exam, Word Fluency Test, Trail Making Test Parts A and B Rey Auditory Verbal Learning Test, Stroop Color and Word Test, and Symbol Digit Modalities Test). We used z scores of changes in the cognitive tests to assess changes in attention, memory, executive function, language, global, and composite cognitive functions in response to the lifestyle and hormonal interventions.
Results: After 6 months, body weight decreased similarly in the LT+Test group and LT+Pbo group (decrease of 9.7 kg vs. 10.3 kg, respectively; P=0.91) whereas testosterone levels increased more in the LT+Test than in the LT+Pbo group (increase of 324 ng/dl vs 88 ng/dl, respectively; P&lt;0.001). Memory z-score increased more in the LT+Test group than in the LT+Pbo group (0.73 vs. 0.39, respectively; P=0.03). Moreover, attention z-score increased more in the LT+Test group than in the LT+Pbo group (0.89 vs. 0.38, respectively; P=0.01). On the other hand, changes in executive function z-score, language z-score, and global z-score did not significantly differ between the LT+Test group and LT+Pbo group (0.45 vs 0.37, 0.34 vs 0.07, and 0.55 vs 0.29, respectively; P=0.13 to 0.56). More importantly, the composite cognitive z-score obtained by averaging all z-scores from each domain increased more in the LT+Test group than in the LT+Pbo group (0.56 vs 0.27; P=0.003).
Conclusion: These findings suggest that in the specific population of older men with hypogonadism and obesity associated with frailty, testosterone replacement therapy can augment the positive effects on cognition from intensive lifestyle intervention by diet-induced weight loss and combined aerobic and resistance exercise.
MON-708 Characterizing DNA Methylation Signatures in Adipose Tissue from Metabolic Impaired Asymptomatic Individuals
Authors: Escalante-Araiza Fabiola , Barajas-Olmos Francisco , Martínez-Hernández Angélica , Huerta-Ávila Eira , Peschard Vanessa-Giselle , Rodríguez-Ayala Ernesto , Bastarrachea Raul A , Orozco Lorena 
Published: May 2020

Abstract
Obesity remains as a global epidemic characterized by progressive metabolic dysregulation in glucose homeostasis. Along with a genetic association in the development of T2D, epigenetic regulation has been suggested as a significant contributor in altered gene expression. Recent studies have described DNA methylation changes in insulin-sensitive tissues involved in T2D pathogenesis, however epigenetic dynamics on early stages to metabolic alterations is still unclear.
We investigated potential DNA methylation signatures in 34 asymptomatic individuals from the GEMM family study. We compared differentially methylated CpG sites (DMC: B value&gt;0 and delta Beta &gt;|10%|; Infinium EPIC array) from subcutaneous adipose tissue (SCAT) in different groups of individuals according to BMI (kg/m2) and HbA1c (%) levels as follow: Group A Control (C): n=9, 22.0±1.9 kg/m2, 4.8±0.3%; Group B Overweight (OW) with normal HbA1c: n=6, 27.8±1.6 kg/m2, 5±0.2%; Group C Obese (OB) with normal HbA1c: n=6, 34.6±4.2 kg/m2, 5.2±0.2%; Group D Prediabetes (PD): n=7, 31.1±5.7 kg/m2, 5.9±0.2% and Group E T2D: n=6, 30.6±7.3 kg/m2, 7.2±0.9%.
We found 43 overlapping genes with shared pathways in all groups, mainly those related to metabolism and adipogenesis. We also documented particular altered methylated genes, in each group (OW: 386, OB:1005, PD:76 and T2D:189). Pathway enrichment analysis in OB and T2D was mainly related to glucose metabolism, while in OW and PD was NOTCH signaling. All groups displayed a consistent hypermethylation in RARA, ESR1 and NCOR2, well known genes involved in lipid metabolism. Additionally, we describe for the first time, a progression toward hypomethylation in ARHGAP15 and MTAP, related with an impaired metabolic status. Otherwise, analysis of overlapping CpG sites revealed a consistently hypermethylated state in OW (86.42%), OB (86.48%) and PD (51.72%), in contrast with the hypomethylation state (56.3%) observed in the T2D group, previously observed elsewhere (1).
In conclusion, comparison of methylation in SCAT obtained from OW, OB, PD and T2D individuals, display potential pathways and DMC signatures specific in each group. Common novel overlapping genes in global DNA methylation profiles of SCAT, were also observed.
Reference: (1) Barajas-Olmos et al., BMC Med Genet. 2018 Feb 21;19(1):1-8.
Nothing to Disclose: FE, FB, AM, EH, GEMM, ER, RB, LO.
OR31-04 Androgen Signaling Regulates Female Fertility Through Modulation of H3K27me3 in Granulosa Cells
Authors: Roy Sambit , Sinha Niharika , Huang Binbin , Cline-Fedewa Holly , Wang Jianrong , Sen Aritro 
Published: May 2020

Abstract
Androgens play a major role in female fertility and in women’s health, in general. However, the underlying mechanism and downstream physiological targets of androgen are not fully understood. Androgen actions are mediated by “nuclear” transcriptional signals or “extra-nuclear” kinase actions. Primary androgen receptor (AR) target genes are those at which AR occupies a genomic androgen response element (ARE) and regulates gene transcription. Subsequently, androgens also regulate other genes in an AR-ARE independent fashion, involving membrane-initiated androgen signaling. In the last couple of years, we have reported that androgens may also influence gene expression through histone modifications. We have found that H3K27me3 (tri-methyl lysine 27 histone3) is a downstream target of androgen actions. H3K27me3 is a gene silencing mark, regulated by Enhancer of Zeste Homologue 2 (Ezh2), a histone methyltransferase that promotes tri-methylation of K27 and androgens inhibit the expression and activity of Ezh2. In this study, we report that androgens also regulate the expression of a histone demethylase called Jumonji domain containing protein 3 (JMJD3/KDM6B), that is responsible of removing the H3K27me3 mark. We find that in granulosa cells (GCs), androgen through the PI3K/Akt pathway, in a transcription-independent fashion, increases hypoxia-inducible factor 1 alpha (HIF1α) protein levels, which in turn induce JMJD3 expression. ChIP studies reveal increased binding of HIF1α on the JMJD3 promoter region with DHT treatment. To understand the global impact of androgens on ovarian gene expression and the contribution of androgen-induced decrease of H3K27me3, we have performed RNA-seq and ChIP-seq analysis with H3K27me3 antibody in primary mouse GCs treated with DHT or vehicle. Results show 190 significantly differentially expressed genes in DHT treated sample vs vehicle, out of which 129 and 61 genes were significantly up- and downregulated, respectively. Moreover, comparison of the RNA-seq and ChIP-seq data reveals that a number of upregulated genes have significantly lower H3K27me3 enrichment in the enhancer and/or promoter region in the DHT treated samples vs vehicle. This establishes that in the ovary, androgen-induced modulation of H3K27me3 mark through regulation Ezh2 and JMJD3 expression/activity is an important regulatory mechanism for ovarian gene expression. To delineate the physiological importance of JMJD3 in normal ovarian function and female fertility, we have also developed a GC-specific JMJD3 knockout mice. We find that GC-specific JMJD3KO mice are sub-fertile with longer estrous cycles and dysregulated follicular development. This phenotype is very similar to GC-specific ARKO mice. Thus, we propose that one of the critical actions of androgens in regulating follicular function and female fertility is through the regulation of JMJD3 expression.
SUN-654 Dynamic and Regional Variation of Pancreatic Innervation in Diabetes
Authors: Alvarsson Alexandra , Rosselot Carolina , Jiménez-González Maria , Li Rosemary , Tzavaras Nikolaos , Wu Zhuhao , Stewart Andrew F , Garcia-Ocaña Adolfo , Stanley Sarah A 
Published: May 2020

Abstract
Background: The pancreas is a highly heterogeneous organ, with regional anatomical, developmental and functional differences. The endocrine pancreas is densely innervated, and neural signals play a significant role in glucose regulation by modulating pancreatic hormone release. However, relatively little is known about the anatomical relationships between islets and nerves across the whole pancreas. Since thin filamentous structures, such as nerves, are difficult to quantify and trace over large volumes using thin section histology, there is a need for high resolution imaging and rendering of intact pancreatic tissue in 3D. Aim: To use optical clearing, whole organ imaging, and 3D rendering to quantify islets and innervation across the whole pancreas in healthy mice, in two mouse models of diabetes, and in pancreatic samples from nondiabetic and diabetic human donors. Methods: Whole-mount staining and clearing was performed using iDISCO+ to quantify innervation, defined by the neuronal marker NF200, and beta cells in pancreata from C57Bl/6 mice, non-obese diabetic (NOD) mice, streptozotocin (STZ)-treated mice, and in pancreatic samples from nondiabetic and diabetic human donors. Z-stacked optical sections were acquired with an Ultramicroscope II at 4x or 12x magnification. Imaris was used to create digital surfaces covering the NF200+ innervation and islets to automatically determine innervation density and islet/nerve interactions. Results: Beta cell volumes were 1-4% in the human pancreas, and 1-2% in the healthy mouse pancreas, with regional variations in islet volume and insulin intensity. There were also significant differences in islet biology between the diabetes models. Innervation of the endocrine pancreas was significantly enriched compared to the surrounding exocrine pancreas, with regional variation. Islets were closely associated with pancreatic innervation and decreased in size with increasing distance from nerves in both mouse and human pancreatic tissue. Innervated islets were relatively preserved in models of diabetes. Finally, islet innervation and expression of neural markers were higher in human samples from diabetic patients and in mouse models of diabetes, with temporal and regional differences. Conclusions: 3D imaging and unbiased analysis across the whole pancreas provides comprehensive measurement of pancreatic nerve volumes and distribution. It allows detailed analysis of the anatomical relationship between nerves and islets, and reveals a close association that is maintained across species. The relative enrichment of innervated islets in diabetes and dynamic changes in islet innervation during the development of diabetes suggest further work is needed to examine the role of pancreatic nerves in preserving and protecting beta cells.
OR02-03 Role of Phosphorylated Gonadotropin-Regulated Testicular RNA Helicase (GRTH/DDX25) in the Regulation of Germ Cell Specific MRNAs in Chromatoid Bodies During Spermiogenesis
Authors: Anbazhagan Rajakumar , Kavarthapu Raghuveer , Dufau Maria L 
Published: May 2020

Abstract
Gonadotropin-regulated testicular RNA helicase (GRTH/DDX 25) is a member of the DEAD-box family of RNA helicases which play an essential role in spermatogenesis. There are two species of GRTH, the 56 kDa non-phospho and 61 kDa phospho forms. Our early studies revealed a missense mutation (R242H) of GRTH in the Japanese azoospermic men which resulted in the lack of phospho-GRTH (pGRTH) in in vitro studies. GRTH knock-in (KI) mice with insertion of the human mutant GRTH gene show loss of the cytoplasmic 61 KDa phospho-species with preservation of the non-phospho nuclear form. KI mice are sterile, lack elongated spermatids and spermatozoa with arrest at step 8 of round spermatids (RS) which contain chromatoid bodies (CB) markedly reduced in size. CB is a non-membranous, cytoplasmic organelle present adjacent to the nucleus of RS, where mRNAs bound to GRTH transported from nucleus to cytoplasmic sites are temporarily stored, translationally repressed for later transport to polyribosomes for translation at specific stages of spermiogenesis. Owing to the specific function of CBs and importance of pGRTH in spermatid elongation, CBs isolated from germ cells of WT and GRTH KI mice were used for subsequent experiments. CBs isolated from GRTH KI mice are smaller, highly condensed and lack the nuage texture of CBs in WT mice. We observed the absence of pGRTH in CB of round spermatids of GRTH KI mice. Also, MVH protein (recognized CB marker protein) was decreased in the CB of GRTH KI mice. Expression of genes related to spermatid regulation, chromatin compaction, remodeling (TP1 and 2, PRM1 and 2, GRTH, TSSK6, HMG2, GCNF, RNF8, TDRD 1, 6, 7 and 9) analyzed by qPCR were markedly reduced in the CB of GRTH KI mice compared to WT. No change was observed in the expression of bromodomain mRNAs and protein, indicating that pGRTH does not participate in the translational regulation of this protein class at the level of this organelle. Notably, mRNAs of TP2, PRM2 and GRTH which associated with GRTH protein were co-localized with MVH protein in the CB. This indicated the relevance of GRTH as a binder/transport protein of key chromatin remodelers for ensuring their mRNA repression/stability within the CB. In addition, GRTH binding to genes essential for spermatid development and regulation (TP1 and 2, PRM1 and 2, GRTH, TSSK6, RNF8 and GCNF) were also found to be markedly decreased in the CB KI mice. These results demonstrate the importance of pGRTH in the maintenance of biochemical composition/structure of the CB and role in spermatid regulation, chromatin compaction, spermatid development and completion of spermatogenesis.
SUN-589 MED1 Is a Lipogenesis Coactivator Required for Postnatal Adipose Tissue Expansion
Authors: Jang Younghoon , Tran Nhien , Ge Kai 
Published: May 2020

Abstract
Mediator is a multi-subunit transcription coactivator complex that controls gene activation by connecting enhancer-binding transcription factors (TFs) with RNA polymerase II (Pol II). The Med1 subunit of Mediator is required for PPARγ-stimulated conversion of mouse embryonic fibroblasts to adipocytes in culture. However, it has remained unclear whether MED1 is required for adipose tissue development in vivo. Using Med1 conditional knockout mice, here we report that MED1 is required for postnatal adipose tissue development/expansion, a process that involves dramatic induction of lipogenic enzymes such as SCD1. Mice with Myf5-Cre-mediated MED1 knockout in progenitor cells show normal embryonic development of brown adipose tissue (BAT) but become runts with reduced BAT and white adipose tissue (WAT) after birth. Furthermore, mice with Adipoq-Cre-mediated MED1 knockout in adipocytes show reduced accumulation of lipid droplets and impaired induction of SCD1 in BAT and WAT. These mice display severe lipodystrophy, cold intolerance, and insulin resistance. In culture, Med1 is dispensable for the induction of adipogenesis markers including PPARγ and C/EBPα in the early phase of preadipocyte differentiation. However, Med1 is required for SCD1 induction, lipogenesis and lipid accumulation in the late phase of adipogenesis. Our preliminary data suggest that MED1 controls the induction of SCD1 by facilitating the recruitment of Mediator and subsequently Pol II to active enhancers bound by the lipogenic TF ChREBP. Taken together, our ﬁndings indicate that MED1 is a lipogenesis coactivator required for postnatal adipose tissue expansion.
SUN-364 Enthesophytes Are a Common Feature of FGF23-Mediated Hypophosphatemia Due to Tumor-Induced Osteomalacia
Authors: Gafni Rachel Ilana , Spector Ellen J , Hartley Iris Ruth , Redd Bernadette , Mitnik Gabriela Lopez , Collins Michael T 
Published: May 2020

Abstract
Background: Tumor-induced osteomalacia (TIO) is a rare cause of FGF23-mediated hypophosphatemia in which mesenchymal tumors produce ectopic FGF23, leading to renal phosphate wasting, decreased 1,25-dihydroxy-vitamin D and hypophosphatemia. Clinical features include muscle weakness, fractures and bone pain. Entheses are sites where tendons, ligaments, fasciae and joint capsules attach to bones. Calcifications in entheses, called enthesophytes, are frequent in adults with X-linked hypophosphatemia (XLH), the most common genetic form of FGF23-mediated hypophosphatemia. One study reported 68% of XLH patients having enthesopathies at an average of 18 different insertion sites per person (Polisson, NEJM, 1985). However, the prevalence of enthesophytes in patients with TIO is not known.
Methods: Skeletal surveys of 66 patients with TIO were reviewed by a single radiologist for the presence of enthesophytes, which were then grouped into the following sites: occiput, axial, upper extremities, pelvis/femur, tibia/fibula and feet. The data presented are from the 59 patients (33 men, 26 women) for whom near-complete skeletal surveys were available; feet radiographs were not available in 9 subjects. Descriptive statistics and regression analyses were performed, including analyses of concurrent intact FGF23 and phosphate levels. Data are presented as mean ± SD.
Results: At the time of the skeletal survey, the age of the subjects was 48.7 ± 14.4 years; 78% were over 40 years. The estimated duration of TIO was 6.6 ± 5.3 years. Mean phosphate level was 1.7 ± 0.5 mg/dL (normal 2.5-4.5 mg/dL) and intact FGF23 was 743 ± 1213 pg/mL (normal &lt; 50 pg/mL). Enthesophytes were identified in 51/59 patients (86.4%) with a mean of 4.5 ± 3.7 enthesophytes per patient (range 0-14). The most frequently affected site was the feet (35/50, 70%) followed by occiput (30/59, 51%), pelvis/femur (28/59, 48%), axial (22/59, 37%), upper extremities (18/59, 31%), tibia/fibula (18/59, 17%). In many subjects, more than one enthesophyte was seen within each region - the total number of enthesophytes in the cohort were: feet – 84, pelvis/femur -74, upper extremities – 40, occiput – 30, axial – 23, tibia/fibula – 16. Multiple linear regression demonstrated a significant positive relationship between number of enthesopathies with age and duration of TIO (p &lt; 0.001). Intact FGF23 and phosphate did not significantly correlate with enthesophyte number.
Conclusions: Similar to XLH, these data demonstrate that enthesopathies are a common feature of tumor-induced osteomalacia, increasing with both age and duration of disease. The underlying mechanism of enthesophytes in the general population is unclear and may be related to mechanical forces and/or inflammation. The additional factors of chronic hypophosphatemia and elevated FGF23 likely contribute to this mechanism.
SUN-283 Lithium Induced Partial Nephrogenic Insipidus: An Unusual Presentation
Authors: Akagi Roberto Kenji Mitsui , Olajide Omolola Bolaji 
Published: May 2020

Abstract
Background: Diabetes insipidus (DI) is characterized by hypotonic polyuria and polydipsia. Nephrogenic DI is the result of an inadequate response of the kidneys to arginine vasopressin (AVP), either due to hereditary causes or acquired from various drugs, most commonly lithium. Clinical case: A 30 year old male with past medical history of Hashimoto’s thyroiditis, severe mental impairment was admitted to the hospital for abdominal pain. Medical history was difficult to obtain since the patient was nonverbal. His thyroid function tests, electrolytes including sodium, potassium were all normal on admission. During the hospital course, he was made NPO for both an upper and lower GI endoscopy. He notably had a mild increase in sodium level with subsequent improvement after restitution of diet. Two days after the procedure, he was found to be drinking out of the toilet and also attempting to drink from the urine jug. Due to this odd behavior, he had to be put on physical restraints. The day after, he was found to have a serum sodium of 158 mEq/L. Hypotonic saline was started and urine output was notably ranging from 6-10 L/day with a negative balance. Desmopressin (DDAVP) test was done with a resultant urine osmolality of 170, 237, 257 and 264 mOsm/kg after 30, 60, 90 and 180 mins. Subcutaneous DDAVP was started with some improvement of serum sodium and decrease in urine output. Endocrinology service was consulted for the evaluation of hypernatremia and polyuria. Initial review of his medications did not show any potential cause of DI. Further inquiry of his prior medication history revealed that he had taken Lithium for over 10 years and stopped 4 months prior to the admission due to polyuria. After excluding other causes of polyuria, with partial improvement of urine osmolality to a level &lt;300 mOsm/kg and a clinical history of prior Lithium therapy, a diagnosis of partial nephrogenic DI was made. Patient was started on HCTZ and given adequate water intake with subsequent improvement of his serum sodium back to normal. Conclusion: Nephrogenic DI due to Lithium use usually recovers after treatment is stopped but could be persistent for several years after. In our case, the patient compensated for his polyuria with increased water intake. When his water intake was restricted due to physical restraints, polyuria and subsequently hypernatremia became evident. A thorough medication history including past drug use is essential as it can help unravel the offending agent which was Lithium in our case. There needs to be an increased awareness that the effect of Lithium in the kidneys could be persistent even after stopping the drug for up to many years. References: Thompson CJ. Persistent nephrogenic diabetes insipidus following lithium therapy. Scott Med J 1997; 42:16-17.