bluuebunny/crossref_metadata_2025_split huggingface dataset:
MON-477 Anti-Streptavidin Interference in Multiple Hormones Immunoassays: Case Report
Authors: Brito Luciana Pinto , Lenczuk Aline , Leopoldino Luciana , Lando Valeria Samuel , Mendonca Berenice Bilharinho , Lima-Valassi Helena 
Published: May 2020

Abstract
Automated immunoassays are the most commonly current methods used in clinicial laboratory to hormone tests, due short turnaround time and high specificity and sensibility. However immunoassays are not free from interferences. Several immunoassays use biotin-streptavidin interaction to anchor antigen-antibody complexes in solid phase and so are susceptible to biotine intake or anti-streptavidin antibody. In sample with these interferents, the results in competitive assays are falsely high and in non-competitive assays are falsely low. OBJECTIVE: To report a case of multiple hormones immunoassay interference by an anti-streptavidin antibody. MATERIAL AND METHODS: A 11-years-old boy with child obesity, and no other symptoms, had discordant laboratory results with markedly elevated free T4, total T4 and total T3 with normal TSH. Repeated measurements in the same sample and in a new collected sample confirmed the initial results and revealed low PTH and high TRAb without hypocalcemia or hyperthiroidism clinical pictures. All the parameters were measured on the automated Cobas e602, Roche plataform.The patient and your family denied biotin ingestion, so anti-streptavidin interference could explain the multiple interference.The serum was pre-incubated with streptavidin microparticles during an hour and centrifuged for 10min (3000 rpm) to removed the analytical interference. RESULTS: The results of post-incubation with microparticles were all in the normal range. Three control samples were not affected by the incubation. Since luminescence is inversely proportional to analyte concentration in competitive assays, low signals lead to falsely high levels (like in free T4, total T4, Total T3 and TRAb in our patient). The reverse occurs with immunometric assays, in which low signals result in falsely reduced values (like in PTH in our patient). CONCLUSION: In literature, streptavidin antibodies seem to be a rare occurrence. In samples with biotin or anti-streptavidin suspection, pre-incubation with streptavidin microparticles is a simple and effective procedure to remove this interference.
SAT-200 Clinical Dilemmas of Adrenal Incidentaloma, Is Change in HU Clinically Significant?
Authors: Niaz Azra , Jun John , Ali Sophia Y 
Published: May 2020

Abstract
Introduction
There has been a recent increase in the incidence of adrenal incidentaloma (AI), defined as an adrenal mass that’s unintentionally discovered on imaging obtained for an indication other than suspected adrenal pathology, mainly because of increased availability and improved quality of imaging modalities.
Differentiation based on malignant vs benign and secretory vs non secretory becomes the most important question on finding such mass. Another important question is long term follow up of these AI. We present a case that highlights the importance of follow up due to the potential of AI to increase in size and density over 3 year.
Case
A 58 years old male with a history of ESRD secondary to hypertension, controlled hypertension, CAD s/p CABG was seen for a right 1.9 cm AI (found in 11/2016 on noncontrast CT abdomen) with low attenuation of approximately 10 Hounsfield units (HU).
He was asymptomatic, normal vitals, and an unremarkable physical exam. Hormonal work up was negative for hyperaldosteronism and Cushing syndrome.
About 1 year later, repeat CT abdomen reported an unchanged right, 2.0 x 1.7 cm adrenal mass, 22 HU. Hormonal work-up was not completed and patient was lost to follow up.
Two years later, repeat CT abdomen reported increase size of AI to 2.7 cm and increase in attenuation of 24 HU precontract which increased to 55 HU on the immediate postcontrast exam and 55 HU on 13-minute delayed images. Hormonal work up was negative for Cushing syndrome and hyperaldosteronism. Plasma metanephrine was 0.50 with elevated plasma normetanephrine of 3.85.
In light of being a renal transplant patient and due to increase in size, change in HU, and significantly elevated normetanephrine levels patient was referred for right adrenalectomy.
Discussion
Change in adrenal mass size is known to be a significant predictor of malignant potential. An unenhanced attenuation value &lt;10 HU is characteristic of a lipid-rich adenoma, can predict benign adrenal adenoma with 98% specificity, and has a very low likelihood of a pheochromocytoma.
European Society of Endocrinology recommends if the AI is homogenous, &lt; 4 cm, with a density ≤ 10 UH, no imaging follow up is recommended and biochemical testing for ruling out pheochromocytoma is indicated in adrenal tumors with an unenhanced attenuation value &gt;10HU. However, the AACE/AAES guidelines recommends AI ≥ 1cm and less than 4cm, repeat imaging with noncontrast CT should be performed at 3–6 months and annually for 1-2 years and if the mass grows or becomes hormonally active then adrenalectomy should be performed.
Conclusion
There are no prospective studies of the optimal frequency and duration of follow up for AI. Considering the increasing incidence of AI, further studies on the clinical significance of change in HU, such as risk of developing pheochromocytoma, along with standardized international guidelines would be helpful for clinicians in managing patients with AI.
SUN-736 Knockout of Membrane Androgen Receptor ZIP9 Results in Reduced Female Fecundity and Abnormal Egg Activation in Zebrafish
Authors: Converse Aubrey , Thomas Peter 
Published: May 2020

Abstract
Recently, our research group cloned and characterized a putative membrane androgen receptor from teleost ovarian tissue that was found to be homologous with the zinc transporter protein ZIP9 (Slc39a9). To date, ZIP9 is the only zinc transporter that is known to be ligand activated or possess steroid receptor activity. Since the discovery of its androgen receptor activity, ZIP9 has been found to mediate androgen actions in a variety of tissues including teleost ovarian follicle cells, human cancer cell lines, and murine Sertoli cells. However, ZIP9 has not been examined in an in vivo model so the precise physiological functions of this receptor remain unclear. A ZIP9-mutant strain of zebrafish was developed using a CRISPR-Cas9 system in order to examine the role of the protein in teleost reproduction. While ZIP9-mutant males had similar breeding occurrence and fertilization rates to wild-type fish, mutant females exhibited severe reductions in fecundity compared to wild-type fish. ZIP9-mutant females spawn significantly fewer eggs of which a high proportion failed to undergo chorion elevation, a characteristic of normal egg activation. Eggs that showed this failed chorion elevation phenotype had significantly lower fertilization rates and produced larvae that exhibit a high incidence of pericardial/yolk sac edema and reduced growth compared to larvae hatched from wild-type eggs. However, no differences were observed in the proportions of oocytes at later stages of development between ZIP9-mutant and wild-type fish, suggesting the observed phenotypes are not related to abnormal oogenesis. We observed that mature wild-type eggs have numerous cortically located vesicles that are autofluorescent under ultraviolet light and decrease in number when the eggs undergo activation, suggesting they undergo exocytosis during the cortical reaction. While zinc is known to be stored in vesicles that undergo exocytosis in mammalian eggs, the role of zinc in teleost egg activation is currently unknown. In eggs from wild-type fish, we observed an increase in extracellular zinc levels upon egg activation and treatment with a zinc ionophore (zinc pyrithione) significantly reduced the number of eggs that undergo normal chorion elevation when activated. This suggests a role for zinc in zebrafish egg activation similar to that observed in mammals. Of interest, ZIP9-mutant eggs that did not undergo chorion elevation had significantly smaller vesicles than those found in wild-type fish eggs. This abnormal vesicle morphology and failure to undergo chorion elevation suggest a role of ZIP9 in egg activation. Additional insight into the role of zinc in zebrafish egg activation and the mechanism by which ZIP9 disruption leads to abnormal cortical vesicles and egg activation will help determine if ZIP9 plays a role in zinc transport and flux in zebrafish eggs during activation.
OR04-04 Identification of a Novel Transcriptional Regulator of Metabolic Disease in Circulating and Central Myeloid Cells
Authors: Sweet David R , Vasudevan Neelakantan T , Fan Liyan , Booth Chloe E , Keerthy Komal S , Jain Mukesh K 
Published: May 2020

Abstract
Derangement in systemic metabolic homeostasis is tightly associated with widespread activation of resident and circulating immune cells, a phenomenon known as ‘metaflammation’. Numerous studies have explored the role of tissue resident and circulating macrophages in contributing to metaflammation, obesity, and their sequelae; however, there is a dearth of information regarding targetable transcriptional regulators of the genesis and persistence of metabolic disease. Here, we identify myeloid Krüppel-like factor 2 (KLF2) as a novel regulator of metabolic disease. Previous reports demonstrate that KLF2 serves as a critical regulator of myeloid cell quiescence and is downregulated in numerous acute and chronic inflammatory states. Specifically in the context of chronic metaflammation, we note that KLF2 expression is decreased in circulating immune cells of obese patients and in adipose tissue macrophages of high fat diet (HFD) fed mice, which is consistent with the hypothesis that KLF2 regulates metaflammation. To explore this further, we utilized mice with myeloid cell-specific deletion of KLF2 (K2KO) which exhibit accelerated obesity and insulin resistance. K2KO mice have widespread central (i.e. CNS) and peripheral metaflammation both in the basal and HFD-stimulated states. To discern whether the effect of myeloid deletion of KLF2 on metabolism is due to deletion in microglia in the feeding centers of the hypothalamus or in peripheral immune cells, bone marrow chimeras with head shielding were created. 50% reconstitution of circulating immune cells with K2KO cells in wildtype (WT) mice was sufficient to maintain the metabolic disease phenotype, while mice with K2KO microglia + WT circulating cells had only slightly improved outcomes compared to K2KO mice. Conversely, ablation of microglia in K2KO mice using PLX5622 formulated in HFD also successfully attenuated the aberrant feeding behavior, weight gain, and glucose dyshomeostasis seen in K2KO mice. Together, these data demonstrate a role for loss of KLF2 in hematopoietic and CNS resident cells in causing metabolic disease. Given that myeloid KLF2 expression decreases under metabolic stress in WT mice and humans, we sought to explore whether maintenance of KLF2 expression in these cells would be protective against diet-induced metabolic disease. Indeed, mice with myeloid-specific overexpression of KLF2 demonstrated a markedly improved metabolic phenotype when challenged with HFD, providing evidence that targeting KLF2 expression in myeloid cells may prove to be a therapeutic option against metaflammation.
MON-588 Anthropometric Parameters, Body Fat Percentage and Metabolic Profile in Sarcopenic Women with Recommendation for Bariatric Surgery
Authors: Carvalho Nara Nóbrega Crispim , Limeira Caio Chaves de Holanda , Borges Heloisa Calegari , Lima Débora Nóbrega , Mesquita Patricia Nunes , Rego Gitana Silva , Holanda Narriane Chaves Pereira , Martins Vinícius José Baccin , Filho João Modesto , Pimenta Flávia Cristina Fernandes , Alves José Luiz de Brito 
Published: May 2020

Abstract
INTRODUCTION: Sarcopenia (SARC) is a musculoskeletal disorder that predisposes several complications, including metabolic ones. Obesity also provides higher risk for metabolic complications, however, there is lack of evidences regarding the association of obesity with SARC on metabolic parameters in non-elderly individuals. OBJECTIVE: To evaluate anthropometric parameters, body fat percentage (BFP) and metabolic parameters in women with and without SARC preceding Bariatric Surgery (BS). METHODS: A cross-sectional study involving 60 obese women in the outpatient care in a public Brazilian University Hospital between March to September 2018. Body composition was given by bio-impedance (inbody-370), multifrequency (5, 50, 250Hz) with 12 hours fasting, dominant Handgrip Strength (HS) was evaluated by Jamar dynamometer (3 measurements; 30 sec interval). Were also evaluated fasting blood glucose, HbA1c, homeostatic model assessment-insulin resistance (HOMA-IR), total cholesterol (TC), low density lipoprotein (LDL), high density lipoprotein (HDL), triglycerides and high-sensitive C-reactive protein (hs-CRP). SARC was defined by the association of a low muscle mass index (weight-adjusted appendicular skeletal muscle mass: ASMM/weight x 100%) and decreased HS, using as cutoff points the smallest quintile for each variable. Data were expressed as mean ± standard deviation and independent t-test was used for comparison between groups. Statistics were made by SPSS software, 20th version (IBM Corp., Armonk, NY). RESULTS: The mean age, weight, body mass index and BFP of sarcopenic and non-sarcopenic women were: 40.75 ± 11 x 39.23 ± 8.92 years old (p=0.665), 102.93 ± 9.58 x 109.19 ± 14.25 Kg (p=0.237), 44.88 ± 2.7 x 42.24 ± 4.79 Kg/m2 and 54.12 ± 1.11 x 51.44 ± 3.43% (p=0.052), respectively. Regarding the laboratory parameters of women with and without SARC: fasting blood glucose 89.25 ± 14.48 x 98.40 ± 27.48 mg/dL (p=0.359), HbA1c 5.83 ± 0.33 x 6.21 ± 1.18% (p=0.185), HOMA-IR 3.61 ± 1.28 x 5.31 ± 4.74 (p=0.160), TC 170.87 ± 39.36 x 180.82 ± 34.51 mg/dL, LDL 94.67 ± 26.63 x 105.60 ± 30.85 mg/dL, HDL 53.37 ± 18.50 x 50.84 ± 10.32 mg/dL, triglycerides 114.12 ± 38.84 x 127.30 ± 75.04 mg/dL and hs-CRP 8.51 ± 6.50 x 7.51 ±6.52 mg/L (p=0.792). CONCLUSION: Women with SARC and recommendation for BS when compared to women without SARC had similar anthropometric, metabolic and BFP parameters.
OR19-06 Sepsis-Induced Critical Illness in Mice Alters Key Regulators of ACTH Production and Secretion Within the Anterior Pituitary Gland
Authors: Téblick Arno , Perre Sarah Vander , Langouche Lies , Van den Berghe Greet 
Published: May 2020

Abstract
Introduction
Critical illness is hallmarked by high plasma cortisol without elevated ACTH. This is partly explained by low cortisol-binding proteins and reduced binding affinity and by suppressed cortisol metabolism, 3 peripheral drivers of increased free cortisol (corticosterone (CORT) in rodents).1 We hypothesized that lack of elevated plasma ACTH is explained by CORT-induced feed-back inhibition either due to suppressed CRH or AVP pituitary signaling or to direct glucocorticoid receptor (GR)-mediated action within corticotrophs. To test this hypothesis, we documented alterations in ACTH precursors and key enzymes controlling ACTH maturation and secretion in a mouse model of sepsis-induced critical illness.
Methods
C57Bl/6 mice were randomly allocated to a healthy control group or to 4 critically ill groups sacrificed after increasing illness duration (30 hours (H), 3 days (D), 5D or 7D). Critical illness was induced by sepsis brought about by cecal-ligation and puncture followed by fluid-resuscitation and antibiotics treatment. The study was continued until 15 surviving animals per time cohort were reached (n=120).
We quantified pituitary pro-opiomelanocortin (POMC) gene/protein expression and POMC plasma concentrations, pituitary POMC intracellular trafficking and cleavage via intracellular POMC sorting/trafficking receptor Carboxypeptidase E (CPE) and prohormone convertase 1 (PC1/3) gene/protein expression. Gene expression of Annexin A1, an inhibitor of mature ACTH secretion, was quantified as marker of GR-mediated CORT-induced feedback inhibition at corticotroph level.
Results
Plasma CORT concentrations were median 3-fold increased during critical illness (p&lt;0.001 for all time cohorts) in the face of normal (for 30H, 3D and 5D cohorts) to low (7D time cohort; p=0.01) plasma ACTH concentrations. Plasma POMC concentrations were higher in critically ill than in control mice (p=0.05). POMC gene expression (but not protein, P=0.8) was a median 55% higher in critically ill mice than in controls (p&lt;0.05 for all time cohorts). In contrast, pituitary mature ACTH protein concentration was median 61% lower in critically ill than in control mice (p&lt;0.01). CPE gene expression was only increased in 30H time cohort (p&lt;0.001). PC1/3 gene and protein expression were positively correlated (R2 0.1; p=0.001) and were reduced (by 37% and 43%, respectively) during the entire course of critical illness (p&lt;0.01). Annexin A1 gene expression was increased during critical illness (p&lt;0.05 for all time cohorts).
Conclusion
Suppressed CRH or AVP signaling and GR-mediated action within corticotrophs explained lack of elevated plasma ACTH in critical illness, as indicated by impaired POMC processing and ACTH maturation. However, increased POMC gene expression suggests ongoing corticotroph activation, the driver of which needs to be identified.
1. Teblick A et al. Nat Rev Endocrinol 2019
MON-018 Fertility Rates in Rats Characterized by Increased Hypothalamic CRH Secretion
Authors: Zoumakis Emmanouil 
Published: May 2020

Abstract
Certain strains of rats are characterized by hyperactive Hypothalamic-Pituitary-Adrenal axis responses to stress, increased hypothalamic Corticotropin-Releasing Hormone (CRH) production and decreased fertility rates. Activation of the HPA-axis and CRH secretion has been associated with suppression of the Hypothalamic-Pituitary-Ovarian axis primarily as a result of glucocorticoids. Here we examined the hypothesis that Fischer rats have decreased fertility rates because of hypothalamic CRH hypersecretion. Antalarmin, a CRH receptor type 1 antagonist, is known to suppress adrenocorticotropin hormone secretion and other CRH receptor type 1-mediated responses. Adult female Fischer rats were injected with antalarmin or placebo, twice a day, for 16 days. Mating was evidenced by the presence of spermatozoa in the vaginal smear performed every morning. After 16 days, 20% of rats (20%) treated with placebo became pregnant and 55% rats treated with antalarmin became pregnant. We have previously reported that administration of antalarmin after the first day of pregnancy does not affect blastocyst implantation in Fischer rats. Our data suggest that antalarmin improves fertility rates in Fischer rats by antagonizing the direct antireproductive role of hypothalamic CRH.
SAT-032 Follistatin-Like 3 (FSTL3) and Early Postnatal Testicular Development
Authors: Mejia Randy Ballesteros , Xia Dong , O’Shaughnessy Peter , Mukherjee Abir 
Published: May 2020

Abstract
Activin, a TGFβ family ligand, induces Sertoli cell (SC) proliferation during the early postnatal stage in mice. Overexpression of activin, however, leads to the disruption of spermatogenesis. FSTL3, a glycoprotein inhibitor of activin, is highly expressed in the testis and its expression is induced by activin creating an inhbitory feedback loop. FSTL3 deletion in mice is therefore expected to produce a mouse model of increased activin action. Contrary to overexpression of activin, however, as we have shown before, global deletion of FSTL3 in mice results in increased numbers of SC and increased sperm production in older males. Stereological analyses show that although the overall number of SC increases with age in both genotypes during pre-pubertal stages, increase in SC numbers is significantly higher in FSTL3 KO males. Here we show our transcriptomic analyses of WT and FSTL3 KO mice at 3d and 8wk. mRNA sequencing data showed that more than 1000 genes are diffferentially regulated between WT and FSTL3 KO at 3d. There is a much lower number of genes differentially expressed at 8wk. Among several canonical pathways that are altered at 3d in FSTL3 KO mice compared to WT we investigated the “Sertoli-to-Sertoli cell Communication” pathway. We found increased expression of junction proteins, including those that are involved in the blood testis barrier (BTB) as well as earlier establishment of BTB in FSTL3 KO testis. Without the BTB, preleptotene spermatocytes cannot progress through the spermatogenic programme. Importantly, we found accelerated SYCP3 organisation, indicative of early entry into meiosis, concomitant with early establishment of BTB. To directly address whether FSTL3 deletion can induce SC proliferation we knocked down FSTL3 expression in the TM4 SC line. We found increased PCNA expression with FSTL3 siRNA compared to control siRNA transfection demonstrating FSTL3 impairment does induce SC proliferation. We are currently investigating the mechanisms for early establishment of BTB in FSTL3 KO testes and additional canonical pathways. The results of the study presented suggests that loss of FSTL3 promotes SC proliferation which likely allows establishment of SC-SC interactions including BTB formation earlier than in WT. Given that the difference in size between WT and FSTL3 KO testis persists at older ages, it is possible that the timing of increase in SC number and establishment of BTB has a sustained effect on testicular function later in life.
SUN-733 Analysis of Divergent Long Noncoding RNAs in Estrogen-Regulated Transcription
Authors: Lee Debra , Yang Barabara , Sedano Melina , Choudhari Ramesh , Gadad Shrikanth S 
Published: May 2020

Abstract
The role of long noncoding RNAs (lncRNAs) in cancer biology are just beginning to be elucidated and recent studies have shown that they could be therapeutic targets. In a previous study, combining powerful techniques, Global Run-On sequencing (GRO-seq) and subcellular fractionation RNA-seq in breast cancer cells identified a large number of estrogen-regulated unannotated long noncoding RNAs. Analysis of gene expression data from hundreds of samples representing 13 different tissue types including both cancer and normal tissue, revealed that many lncRNAs are differentially expressed in various cancers. Furthermore, a large number of lncRNAs are divergent transcripts and show distinct expression patterns across molecular subtypes of cancer. In functional assays, knockdown of selected lncRNA, such as lncRNA67, inhibits the growth of breast cancer cells. Amplified expression of lncRNA67 in luminal-subtype of breast cancer correlates with clinical outcome. LncRNA67 has now been fully annotated (transcription start and stop site, 5’ cap, polyA tail, and exon/intron structure), and cloned. Our preliminary molecular analyses indicate that lncRNA67 plays a critical role in ER-dependent and -independent pathways. Collectively, our results suggest that lncRNAs are an integral component of cancer biology.