bluuebunny/crossref_metadata_2025_split huggingface dataset:
SUN-638 Perinatal DDE Exposure Disrupts Thermogenesis Early in Development
Authors: vonderEmbse Annalise , Tong Jason W , Lein Pamela J , Merrill Michele La 
Published: May 2020

Abstract
Numerous epidemiological studies have identified a positive association of exposure to the endocrine-disrupting pesticide dichlorodiphenyltrichloroethane (DDT) and its primary metabolite dichlorodiphenyldichloroethylene (DDE) with the risk of obesity. Of particular concern is the persistent metabolic dysfunction resulting from early life DDT and DDE exposures, evidenced as obesity, glucose intolerance, dyslipidemia, and impaired thermogenesis in adult rodents. However, little is known about the developmental timing and etiopathogenesis of this long-term DDT-related metabolic phenotype. This study aimed to address these knowledge gaps by evaluating endocrine phenotypes and thermogenic parameters at two postnatal (P) time points, P0 and P12, in C57BL/6J mice. Dams were orally gavaged with p,p’-DDT (1.7 mg/kg body weight [BW]), p,p’-DDE (1.31 mg/kg BW), or organic olive oil (vehicle) daily from gestational day (GD) 11.5-P0 or P5. Infrared analysis was then performed during a cold challenge. We further attempted to rescue the cold-challenged P12 mice by administering the β3-adrenergic receptor (AR)-agonist CL316,243 (CL), a direct stimulator of brown adipose (BAT) thermogenesis. At P0, area under the curve analysis revealed higher body temperatures in both males and females exposed to DDE compared to controls during the 9 min cold challenge. In addition, DDE-exposed females lost body heat at a significantly faster rate than sex-matched controls over the 9 min cold challenge. This occurred without any treatment-related differences in resting glucose, suprascapular BAT weight, or body weight for either sex at P0. To assess BAT-autonomous response to β3-AR stimulation during a cold challenge, P12 mice were exposed to the pharmacological β3-AR agonist, or PBS control in a 2x2 exposure design and suprascapular BAT temperature was evaluated via infrared analysis of the suprascapular surface at 10 min intervals. Blood glucose, BAT weight, and BW remained equivocal across all P12 treatment groups. However, BAT temperature was significantly decreased 10 min after cold- challenge in female P12 mice with perinatal DDE-exposure when compared to sex- and perinatal treatment-matched controls. This DDE effect among P12 female mice was rescued by CL. These data suggest that thermogenic dysfunction consequent to perinatal DDE exposure is detectable during postnatal development, well before the emergence of endocrine phenotypes. The CL rescue of the BAT thermogenic impairments observed after perinatal DDE exposure are consistent with DDE toxicities upstream in the β3-AR nerve circuitry. Additionally, the data highlight the emergence of an early postnatal sex divergence in DDE-related thermogenic dysfunction. We speculate that the effects of DDE on suprascapular BAT heat production result from prenatal alterations in sympathetic circuitry.
SUN-572 Estrogen Synergistically Interacts with Optic Atrophy Protein 1 to Promote Thrombosis
Authors: Souvenir Rhonda A , Renata Alambert , Campbell Robert , Hinton Antentor O , Rondina Matthew T , Abel Evan Dale 
Published: May 2020

Abstract
Thrombosis is a major concern in: premenopausal females on oral contraceptives, menopausal women undergoing hormone replacement therapy, post-menopausal women and transgender individuals receiving estrogen supplementation. The mechanisms linking estrogen exposure with increased thrombosis risk is incompletely understood. Analysis of platelet transcripts in the Framingham cohort identified Optic atrophy 1 (OPA1) expression as being highly predictive of female sex and correlating with increased risk of diabetes and cardiovascular disease. OPA1 regulates mitochondrial fusion, electron transport chain (ETC), complex assembly, cristae morphology and apoptosis. Thus, to determine the functional relationship between platelet OPA1 expression and platelet function in relation to sex steroids we generated mice with platelet specific deletion of OPA-1 protein (pOPA1KO). Male pOPA1KO exhibited structurally and functionally compromised mitochondria with a 50% reduction in mitochondrial DNA and respiration. Male pOPA1KO mice exhibited a prothrombotic phenotype they had: increased agonist-induced activation, shortened time to stable occlusion of the carotid artery as assessed in vivo by (rose Bengal) photochemical injury (~25 min knockout vs ~ 35 min control), and were more prone to develop a thrombus (14/15 knockouts vs. 4/8 controls) following permanent ligation of the inferior vena cava. In contrast, female pOPA1KO mice had normal mitochondrial structure, function and DNA. Both agonist-induced platelet activation and thrombus formation was unchanged in pOPA1KO females. Paradoxically, pOPA-1 KO female mice had increased time to stable occlusion of the carotid artery as assessed by photochemical injury (~75 min Knockout vs ~35 min control). Notably, when platelets from pOPA-1 KO or control males were transferred into females following depletion of endogenous platelets, the reconstituted male platelets acquired the phenotype of female pOPA-1 KO mice. Thus, the time to stable occlusion of the carotid artery following photochemical injury was increased. Similarly, reconstitution of male mice with female pOPA1KO platelets were no longer prothrombotic. Gonadectomized pOPA1KO males had an increased time to stable occlusion and gonadectomized female pOPA1KO no longer exhibited increased time to stable carotid artery occlusion. Eugonadal pOPA1KO male mice treated with estrogen exhibited the pOPA1KO female thrombotic phenotype, with increased time to stable occlusion relative to control males. Additionally, OPA-1 levels were positively correlated with increased platelet aggregation and increased estrogen levels in third trimester pregnant human females. Together, these findings reveal a synergistic interaction between platelet OPA1 levels and estrogen to promote thrombosis.
MON-254 Sympromic Hypogonadism: Co- Existence of Morsier and Klinefelter Syndromes
Authors: romero Fabiola , Arzamendia Sady , Ferreira Dahiana , de Souza Claudia Neves , Lopez Helen , Castellano Barbara , Valinotti Elizabeth , Ayala Alejandro 
Published: May 2020

Abstract
INTRODUCTION: Morsier Syndrome is a rare congenital malformation, characterized by hypoplasia / aplasia of the septum pellucidum and hypoplasia / aplasia of the optic nerves, in addition to pituitary and hypothalamic hormonal deficiencies. Klinefelter Syndrome is a sexual chromosomal genetic alteration, a frequent cause of male hypogonadism, The association of Morsier syndrome and Klinefelter is described below.
CLINICAL CASE
We report he case of a 12 year-old boy with psychomotor retardation and nystagumsho presented at 14 months of age with growth hormone deficiency (low weight and height,) and diabetes insipuidus with hypernatremia of of 159 mEq and low urinary density (less than 1,005).
MRI showed an absence of septum pellucidum, thick right frontal cortical dysplasia with asymmetric appearance of the grooves, small optic chiasma, hypoplastic pitutary gland (3 mm height), compatible with Morsier syndrome.
The physical examination draws attention to tall stature, and long lower limbs, facies with prominent forehead and hypertelorism, gynecomastia and small external genitalia for age.
Hormonal evaluation revealed hypergonadotropic hypogonadism with a 47 XXY karyoteype suggeting Klinefelter syndrome.
CONCLUSION:
We report the first case of Morsier syndrome, associated to Klinefelter syndrome. Both syndromes may present with hypogonadism. However, the diagnosis of klinefelter syndrome was made based on the phenotypic characteristics of this patient including hyeprgonadotroic hypogonadism and abnormal karyotype analysis.
SUN-LB104 Metabolic Inflexibility: Is It a Feature of Obesity or a Characteristic of Metabolically Unhealthy Youth?
Authors: Gebara Nour Y , Kim Joon Young , Bacha Fida , Lee SoJung , Arslanian Silva A 
Published: May 2020

Abstract
Obese individuals have metabolic inflexibility evidenced by diminished fasting fat oxidation and blunted increase in respiratory quotient (RQ) from fasting to insulin-stimulated state. Metabolically unhealthy obese (MUHO) adolescents, unlike their metabolically healthy obese (MHO) peers, have unfavorable metabolic characteristics despite having comparable adiposity. We investigated if metabolic inflexibility is a characteristic of obesity per se or is unique to MUHO compared with MHO youth.
Obese youth (n=188; age 14.1 ± 0.1 yrs [SE]; BMI 33.6 ± 0.4 kg/m2) were divided into 137 MUHO (age 14.1 ± 0.2 yrs; BMI 35.4 ± 0.5 kg/m2) and 51 MHO (age 13.9 ± 0.3 yrs; BMI 29.0 ± 0.7 kg/m2) based on cut points for in vivo insulin sensitivity (IS) [MHO within 1.5 SD and MUHO &lt;1.5 SDs of 72 normal-weight (NW) adolescents’ IS values]. RQ (by indirect calorimetry) at fasting and during a hyperinsulinemic (80mu/m2/min)-euglycemic clamp was measured, and ∆RQ calculated. Body composition (by DEXA), visceral adipose tissue (VAT) (by CT and MRI), hepatic IS (HIS) (calculated from fasting hepatic glucose production by [6,6-2H2]glucose and fasting insulin), adipose IS (ATIS) (calculated from whole body lipolysis by [2H5]glycerol and fasting insulin), and peripheral IS were assessed.
MUHO vs. MHO youth had blunted ∆RQ (0.088 ± 0.004 vs. 0.107 ± 0.007, p=0.035), but MHO was not different from NW (0.098 ± 0.004, p=0.893). Further, MUHO vs. MHO youth had lower HIS (15.3 ± 0.7 vs. 24.3 ± 1.6 (mg/kg/min·uU/mL)-1, p&lt;0.0001) and lower ATIS (9.8 ± 0.5 vs. 22.3 ± 3.1 (umol/kg/min·uU/mL)-1, p&lt;0.0001), but HIS and ATIS were not different between MHO and NW youth (24.3 ± 1.6 vs. 20.8 ± 1.2 (mg/kg/min·uU/mL)-1, and 22.3 ± 3.1 vs. 22.0 ± 1.4 (umol/kg/min·uU/mL)-1, p=ns for both). ∆RQ correlated with HIS (r=0.535), ATIS (r=0.288) and VAT (r=-0.309) (p&lt;0.0001 for all), but not with BMI, BMI Z-scores or % body fat. The differences between MUHO and MHO youth in ∆RQ, HIS and ATIS remained significant after adjusting for % body fat, race, pubertal status and VAT.
The present study reveals that metabolic inflexibility is not a feature of obesity, rather it is a characteristic of MUHO youth who have significantly lower ∆RQ compared with MHO youth, with no difference between MHO and NW youth. Moreover, MUHO compared with MHO youth have worse metabolic profile, represented in lower HIS and ATIS.
SAT-LB58 Molecular Investigation of Recessive Inheritance by Exome Sequencing of Patients With Congenital Hypopituitarism
Authors: Pereira Ferreira Nathalia Garcia Bianchi , Madeira Joao Luiz do Oliveira , Kertsz Renata , Gergics Peter , Benedetti Anna Flavia Figueredo , Biscotto Isabela Peixoto , Fang Qing , Ma Qianyi , Ozel Bilge Ayse , Li Jun , Camper Sally Ann , Jorge Alexander Augusto Lima , Mendonca Berenice Bilharinho , Arnhold Ivo J , Carvalho Luciani Renata Silveira 
Published: May 2020

Abstract
Background: Growth hormone deficiency (GHD) occurs in ~ 1/8000 individuals, and 14% of the patients have mutations in five major candidate genes. However, over 30 genes have been implicated in hypopituitarism. WES (Whole Exome Sequencing) is a promising approach for molecular diagnosis of patients with GHD because it offers the opportunity to screen for all known genes in addition to novel disease gene discovery. Methods: WES was performed for 13 unrelated patients with congenital hypopituitarism born from consanguineous parents. The variants were filtered assuming autosomal recessive inheritance, rare variants in population databases, in silico analysis predicted as deleterious and pituitary and/or hypothalamus gene expression. To determine whether variants in CDH2 that were predicted to be deleterious were functionally significant, L1 fibroblast lines that have no endogenous CDH2 protein were stably transfected with either human wild type or variant CDH2, the transfected cells were labelled with lipophilic dyes, and cell adhesion properties were assessed. Results: Homozygous pathogenic or likely pathogenic allelic variants were found in 2 of the 13 patients. First, a female patient with GH, TSH, ACTH and LH/ FSH deficiencies presenting ectopic posterior pituitary lobe, non-visualized stalk, and hypoplastic anterior pituitary lobe had two homozygous rare variants predicted as deleterious: PLA2G4A p.Asn703Lys and CDH2 c.865G&gt;A (p.Val289Ile). Only CDH2 is known to be expressed in the pituitary, and Pla2g4a null mice have a pleiotropic phenotype without obvious hypopituitarism. The CDH2 variant is rare and classified as deleterious. Sanger sequencing of CDH2 in four family members of the affected proband revealed that the unaffected parents and two unaffected siblings were heterozygous carriers. The effect of the CDH2 variant on cell aggregation was assessed in cell culture. Large cell aggregates formed in cells transfected with wild type CDH2, but cell aggregates were small or absent in cells that were either non-transfected or transfected with the CDH2 variant. Second, a patient with isolated GHD and no MRI abnormalities was identified with a rare, likely deleterious, homozygous GH1 c.171delT (p. Phe 57Leufs*43) variant. He had a sister who died at the age of 5 and had features of GHD. Conclusion: In a cohort of congenital hypopituitarism from consanguineous parents we had 15% molecular diagnosis using WES. We identified a variant in a known gene, GH1 c.171delT and a variant in a novel gene, CDH2 p.Val289I.
SAT-LB97 MiRNA-99a and mTOR2 Mediate Enhanced Endothelial Mineralocorticoid Receptor Signaling-Induced Activation of Sodium Channel and Endothelium Stiffness
Authors: Jia Guanghong , Aroor Annayya R , Habibi Javad , Yang Yan , DeMarco Vincent G , Hill Michael A , Whaley-Connell Adam T , Jaisser Frederic , Jaffe Iris Zamir , Sowers James R 
Published: May 2020

Abstract
In diet induced obesity enhanced endothelial cell (EC) mineralocorticoid receptor (MR) (ECMR) and downstream sodium channel (EnNaC) activity increases oxidative stress and inflammation, thereby promoting vascular stiffness and associated impaired endothelial mediated relaxation. For example, consumption of a Western diet (WD) containing excess fat (46%) and fructose (17.5%) for 16 weeks elevated plasma aldosterone levels and increased vascular MR expression in conjunction with increased endothelial and vascular stiffness in female mice. EC specific deletion of either the ECMR or EnNaC significantly attenuated this diet induced endothelial/vascular stiffness. Emerging information suggests that abnormal expression of miR-99a may be involved in these processes. To this point, we recently observed that aldosterone (10-7 mol/L) causes a reduction in miR-99a that was prevented by the MR antagonist, spironolactone (10µM) in in vitro ECs. By using RNA sequencing, we also demonstrated that ECMR activation reduced arterial miR-99a expression in diet induced obesity. Since the mammalian target of rapamycin (mTOR2)/SGK1 signaling pathway is involved in aldosterone activation of ENaC we then explored the effects of miR-99a on mTOR2 expression. Indeed, miR-99a reduced mTOR2. We further observed that inhibition of mTOR2 with PP242 inhibited EnNaC activity as determined by patch clamping of ECs. Collectively these data suggest that consumption of a WD induced ECMR activation and increased EnNaC activity and endothelial stiffness, in part, by reducing the tonic inhibitory effects exerted by miR-99a on mTOR2 mediated EnNaC activation.
SUN-LB94 Impact of the Gut Microbiome and Renin-Angiotensin-Aldosterone System in Hypertensive Patients With Low-Salt Intake
Authors: Taniguchi Kouki , Nagase Satoshi , Karashima Shigehiro , Kometani Mitsuhiro , Aono Daisuke , Higashitani Takuya , Yoshikura Syohei , Takeda Yoshiyu , Nambo Hidetaka , Yoneda Takashi , Okamoto Shigefumi 
Published: May 2020

Abstract
Salt intake is one of most important environmental factors responsible for triggering the onset of hypertension. Renin-angiotensin-aldosterone system (RAAS) plays a key role in adjusting sodium homeostasis and blood pressure. Recently, the potential role of the gut microbiome (GM) in altering the health of the host has drawn considerable attention. We investigated the impact of intestinal microflora and RAAS in hypertensive patients with low-salt or high-salt intake using an observational study.
A total of 239 participants were enrolled and their GMs and clinical backgrounds examined, including the renin-angiotensin-aldosterone system and inflammatory cytokine levels. On the basis of enterotypes—determined by cluster analysis—and salt intake, the participants were classified into four groups, low salt/GM enterotype 1, low salt/GM enterotype 2, high salt/GM enterotype 1, and high salt/GM enterotype 2.
The prevalence of hypertension was significantly lower in the low-salt intake (low salt/GM enterotype 1 = 47% vs low salt/GM enterotype 2 = 27%, p = 0.04) groups. No significant difference in the prevalence of hypertension was observed for the two GM enterotype groups with high-salt intake (GM enterotype 1 = 50%, GM enterotype 2 = 47%; p = 0.83). Plasma aldosterone concentration was significantly different among the four groups (p &lt; 0.01). Furthermore, the relative abundance of Blautia, Bifidobacterium, Escherichia-Shigella, Lachnoclostridium, and Clostridium sensu stricto was also significantly different among these enterotypes. This suggested in certain individuals (with specific gut bacteria composition) changing dietary habits—to low salt—would be ineffective for regulating hypertension through RAAS. Our findings provide a new strategy for controlling blood pressure and preventing the development of hypertension through restoring GM homeostasis.
SAT-LB122 Characterization and Functional Rescue of a Nephrogenic Diabetes Insipidus Causing S127F Substitution in V2 Vasopressin Receptor
Authors: Hunyady Laszlo 
Published: May 2020

Abstract
Laura Szalai1, András Sziráki1, László Sándor Erdélyi1, András Balla1,2, László Hunyady1,2
1Department of Physiology, Semmelweis University, and 2MTA-SE Laboratory of Molecular Physiology, Budapest, Hungary
The concentrating function of the kidney is important to maintain the water homeostasis of the body. It is regulated by the arginine-vasopressin system through the type 2 vasopressin receptor (V2R). Loss-of-function mutations of V2R in kidney can lead to nephrogenic diabetes insipidus (NDI) which results several symptoms such as polyuria, polydipsia, and hyposthenuria.
In this study, we functionally characterized and investigated the potential rescue of a missense mutation (S127F) of the V2R. We monitored the cellular localization of the S127F mutant V2 receptor using HA-tagged receptors in confocal microscopy experiments. The S127F V2 receptor was detected only in the endoplasmic reticulum but not in the plasma membrane. Flow cytometry measurements revealed that only limited amount mutant receptor can be found on the cell surface compared to the wild type V2R. We also determined the cAMP signaling capability of the mutant receptor with BRET measurements. The S127F receptor was not able to increase the intracellular cAMP levels in response to vasopressin stimulation. Certain ER retention mutations can be rescued by pharmacological chaperones, which cause misfolded mutant receptors to present in the plasma membrane. We examined the effect of tolvaptan (a V2R antagonist) on the S127F V2 receptor. HEK293 cells were transiently transfected with the plasmid of the mutant receptor and after one day the cells were incubated for 18 hours with tolvaptan. After the pretreatment, the cells were exposed to vasopressin, and we were able to detect significant cAMP signal generation of the mutant receptor. We also checked whether the result after tolvaptan pretreatment was due to restored plasma membrane location of the receptor. We were able to demonstrate significant increase of the mutant receptors in the plasma membrane using flow cytometry. We also investigated the effect of MCF14 compound (a cell permeable high-affinity agonist for the V2R) on the mutant receptor and we found that the MCF14 was also capable to restore the cAMP signaling function of the receptor.
This work was supported by the National Research, Development and Innovation Fund (NKFI K116954 and NVKP_16-1-2016-0039).
SAT-LB135 A Novel Cytoplasmic Membrane Estrogen-Mediated Biogenic Signaling Pathway
Authors: Pestell Richard G 
Published: May 2020

Abstract
The estrogen receptor α (ERα) is known to convey both genomic and extra-genomic activities. The extra-nuclear estrogen signaling pathway is thought to involve a membrane-associated estrogen receptor (ERα), which activates PI3-kinase and Akt signaling. Maximal activation of Akt requires S473 phosphorylation. The essential G1-cyclin, CCND1, is a collaborative nuclear oncogene that is frequently overexpressed in cancer. D-type cyclins bind and activate CDK4/6, contributing to G1-S cell-cycle progression. Herein, cyclin D1 was shown to be located in the cytoplasmic membrane of patients with inflammatory breast cancer, human diploid fibroblasts and cancer cell lines (breast, prostate). The extra-nuclear vs. nuclear E2-induced signaling pathways can be distinguished using 17β-estradiol linked to a dendrimer conjugate (EDC), which excludes estradiol from the nucleus. In contrast with the nuclear-localized form of cyclin D1 (cyclin D1NL), the cytoplasmic membrane-localized form of cyclin D1 (cyclin D1CML) was sufficient to induce phosphorylation of the serine threonine kinase Akt (Ser473) and augmented extra-nuclear localized 17β-estradiol dendrimer conjugate (EDC)-mediated phosphorylation of Akt (Ser473). Cyclin D1CML was sufficient to induce G1-S cell-cycle progression, cellular proliferation, colony formation. In contrast with cyclin D1NL, the cyclin D1CML induced transwell migration and the velocity of cellular migration. Together these studies suggest distinct subcellular compartments of cell cycle proteins may convey distinct functions. The major adjuvant therapy for the ~70% of ERα expressing human breast cancer involves anti-estrogen therapy and the ERα/PI3K/Akt complex pathway is hyperactivated in aggressive breast tumors. The non-genomic actions of E2/ERα, mediated via cyclin D1CML may provide an important additional target. References. 1. 2. Casimiro MC et al Mol Endocrinol. 2013;27(9):1415-28. Di Sante, G, Expert Rev Anticancer Ther. 2019 Jun 20:1-19.