bluuebunny/crossref_metadata_2025_split huggingface dataset:
Big data and genomics of Insulin to expand our knowledge
Authors: Cook Taylor , Prokop Jeremy 
Published: May 2023

Physiology research must be guided moving forward by our significant investments represented by deposited data. We hypothesize that integrating these resources, namely our investments into genomics, makes it possible to discover many aspects of well-studied physiological pathways. Within this work, we build a genomics integration for the Insulin gene/protein, which serves as an example of data integrations that can be applied to any physiological system. The INS gene region has 2,076 unique variants from population genetics of hundreds of thousands of individuals (gnomAD, ClinVar, Geno2MP, Avada, dbSNP). Several variants are found near the transcriptional start site, enhancers, and following the INS transcripts that might influence the readthrough fusion-transcript INS- IGF2. This INS- IGF2 transcript splice site is confirmed within hundreds of pancreas RNAseq samples, lacks drift based on human genome sequencing, and has a possible elevated expression due to viral regulation within the pancreas/liver. Moreover, a rare, poorly characterized African-specific variant of INS-IGF2 results in a loss of the stop codon. INS transcript UTR variants rs689 and rs3842753, highly associated with Type 1 Diabetes, are found in hundreds of pancreas RNAseq datasets with an elevation of the 3’UTR alternatively spliced INS transcript. Finally, by combining literature, evolutionary profiling, and structural biology, we map rare missense variants involved in type-1 diabetes, type-2 diabetes, and monogenic/neonatal diabetes that influence Pre-Pro-Insulin translation, Pro-Insulin processing, dimer/hexamer secretory storage, receptor activation, and C-peptide detection for quasi-Insulin blood measurements. Finally, through these complex Insulin pathways and multiple animal model species, we highlight the many genes involved and how environmental factors such as cellular stress may interact with genetics to increase the prevalence of diabetes. Overall, our data integration strategy yields a platform for merging our investments into animal physiological model systems, human clinical genomics, and growing investments into genome sequencing to highlight the many exciting future research needs of physiological pathways. This research was funded by the Gerber Foundation (to JWP), National Institutes of Health (K01ES025435 to JWP), and Michigan State University. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Objective cardiovascular autonomic abnormalities persist over time in patients with Post-Acute Sequelae of COVID-19 (PASC)
Authors: Hira Rashmin , Siddiqui Tanya , Baker Jacquie , Bourne Kate , Ranada Shaun , Soroush Ateyeh , Karalasingham Kavithra , Ahmad Hyeqa , Morillo Carlos , Sheldon Robert , Raj Satish 
Published: May 2023

BACKGROUND: The novel coronavirus (SARS-CoV-2) has negatively impacted the health and economy of the world. While most patients recover, many are suffering from symptoms several months after resolution of the acute illness (“Post-Acute Sequalae of COVID-19” [PASC]). Symptoms include fatigue, light-headedness, and tachycardia/palpitations, similar to symptoms of cardiovascular autonomic disorders such as Postural Orthostatic Tachycardia Syndrome (POTS), Initial Orthostatic Hypotension (IOH), Orthostatic Hypotension (OH), and Inappropriate Sinus Tachycardia (IST). We have previously shown a high prevalence of autonomic abnormalities among PASC patients. We currently do not know if these autonomic abnormalities are chronic and persist in PASC patients over time. OBJECTIVE: We aimed to determine if the autonomic abnormalities seen in patients after COVID-19 infection are persistent or transient, by comparing hemodynamic criterion between baseline and follow-up visits. We hypothesize that patients will recover, and abnormalities will not persist between the visits. METHODS: PASC patients (n=64; F=51; 46±12y) completed baseline visits 411±133 days after their initial COVID-19 infection and follow-up visits 97±9 days later. Patients underwent autonomic function testing with beat-to-beat hemodynamics during 10min supine rest followed by a 10min active stand. Patients were evaluated for hemodynamic criterion for POTS (­­­­↑HR≥30bpm within 10min), IOH (transient ↓SBP≥40mmHg within 15s), OH (↓SBP≥20mmHg within 3min), and IST (supine HR>100bpm). Categorical data were analyzed with a McNemar’s test. RESULTS: 19 (30%) participants met POTS criterion at baseline vs. 15 (23%) at follow-up (p=0.1). 43 (67%) participants met IOH criterion at baseline vs. 37 (58%) at follow-up (p=0.08). The OH criterion was seen in 2 (3.1%) vs 5 (7.8%) patients (p=0.2) and IST was seen in 1 patient at both visits (1.6%). Overall, 48 (75%) patients met the hemodynamic criterion for at least one autonomic disorder at baseline and 43 (67%) patients met at least one criterion at follow-up (p=0.09). CONCLUSION: Many patients with PASC have evidence of cardiovascular autonomic abnormalities, most commonly IOH, followed by POTS. The prevalence of these abnormalities does not decrease over time. These findings suggest patients with PASC are not recovering from autonomic complications due to their COVID-19 infections. Treatment and management strategies for autonomic abnormalities are needed to improve the quality of life for these patients. Funding: Canadian Institutes of Health Research (CIHR) This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Osteopontin Expression is Upregulated in Gonadal Adipose Tissue During Early Menopause Stage in Female Mice
Authors: Hossain Mehorunnesa , Maddie Nicole , Carrillo-Sepulveda Maria 
Published: May 2023

Background: Menopausal women are at high risk of developing atherosclerosis. Reduced ovarian hormone production is a key feature of menopause, and it often leads to increased visceral adiposity and atherosclerosis. Osteopontin (OPN) is an inflammatory cytokine found in high levels in menopausal women, and in human atherosclerotic plaque. While recent studies report that macrophages in male epididymal white adipose tissue are able to produce OPN, less is known about the gonadal white adipose tissue (gWAT) in females. We hypothesize that OPN protein expression is up-regulated in gWAT in early stages of menopause, conferring gWAT as a source of OPN.Methods: To address our hypothesis, a bilateral ovariectomy in a surgical menopausal murine model was utilized. Twelve-week-old female C57BL/6 sexually mature mice were randomized into two experimental groups: Sham group (n=5) whose ovaries remained intact, and OVX group (n=6), who underwent ovariectomies, and were observed for four weeks therafter. At terminal experiments, gonadal white adipose tissue (gWAT) was collected for biochemical analysis. Results: As expected, OVX mice exhibited increased body weights (24.07 ± 0.85 g vs 21.06 ± 0.38 g controls, p<0.05) and increased gWAT mass (13.69 ± 1.421 mg/g vs 9.58 ± 0.55 mg/g control, p<0.05), indicating that early decline in female hormones is able to promote adiposity. Strikingly, greater OPN expression (2.57 ± 0.50 a.u vs. 0.76 ± 0.13 a.u sham group, n=3, p<0.05) was observed in the gWAT from the OVX group. Conclusion: Our results indicate that OPN expression in gWAT is increased during early stages of menopausal status, and suggest that gWAT could be a potential source of high levels of OPN in menopause. Our data suggest that female gWAT is not only an expanded fat depot but a metabolically active tissue contributing to an inflammatory state in menopause by an increased production of circulating cytokines such as OPN, which can ultimately contribute to inflammatory processes, including atherosclerosis. Funding: Diabetes Action to Maria Alicia Carrillo Sepulveda This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Sodium channel protein alpha subunit genomics: building an integrated knowledgebase to translate between family members
Authors: Zieba Jacob , Wilstermann Amy , Hernandez Angel , Prokop Jeremy 
Published: May 2023

The sodium channel alpha proteins consist of ten homologous human proteins (SCN1A, SCN2A, SCN3A, SCN4A, SCN5A, SCN7A, SCN8A, SCN9A, SCN10A, SCN11A), many involved in various human physiological pathways and clinical disorders/syndromes. Many human variants connected to these genes remain uncertain for human disease. To better support the understanding of challenging human variants, we have built an ortholog database of the alpha subunit proteins for the sodium channel. We combined 1,680 vertebrate species sequences of family members, protein modeling, molecular dynamic simulation of lipid embedded protein, screening of human variants (gnomAD, ClinVar, Geno2MP), and analysis of a large number of human RNAseq and single-cell RNA-seq to understand the function of each amino acid in the human proteins and how the amino acids are conserved across family members to contribute to sodium channel functions within diverse cells of human. There are 610 amino acids conserved across the alpha subunit family, representing the most conserved amino acids for each member. These amino acids drive the structural fold, membrane interactions, and channel opening. A cluster of amino acids is conserved between a subset of family members that defines subclusters of alpha subunit proteins. Expression analysis of each member reveals clusters of members co-expressed in various cells, where heart and brain datasets show the specificity of cell types that contribute to genotype-to-phenotype associations of each member. Further, to support variant characterizations, the ortholog alignment data provides a unique ability to discover loss vs. gain of function variants found between members, allowing for known pathogenic variants of one member to aid in the characterization of another member's uncertain variants. For example, a clinical variant of SCN8A linked to seizures was supported as loss-of-function through variants within SCN1A (epilepsy), SCN2A (seizures), SCN4A (Familial hyperkalemic periodic paralysis), and SCN5A (Brugada syndrome). Overall, we show an exciting and promising new tool for ortholog genomic mapping that can support clinical variant insights into physiological outcomes (genotype-to-phenotype relationship), which should be applied to other cellular ion channels and transporters. This research was funded by the Gerber Foundation (to JWP), National Institutes of Health (K01ES025435 to JWP), and Michigan State University. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Transgenerational effects of opioids on cardiovascular outcomes of the rat offspring
Authors: Ahmed Nermin , LORIA ANALIA 
Published: May 2023

Opioid use disorder (OUD) is one of the greatest public health threats in the US. In 2020, more than 69,000 Americans died of a drug overdose involving opioids. Starting in 2014, an alarming third wave is on the rise now caused by synthetic opioids, specifically illicit fentanyl (FEN). OUD among pregnant women is an understudied area related to the opioid epidemic, showing a 5-fold increase incidence of neonatal opioid withdrawal syndrome (NOWS). Currently, the only two FDA-approved medications indicated for medication assisted treatment (MAT) during pregnancy are methadone and buprenorphine. While some general guidelines have been established to place pregnant woman on MAT, the misuse of fentanyl has challenged the current pharmacotherapies, urging for the characterization of pre-clinical models.The aim of this study was to investigate the trasngenerational effects of opioids on the cardiovascular system. We subjected female rats to opioids exposure during pregnancy. We investigated: 1) In the newborns, the morphometrics and withdrawal symptoms; 2) In the adult offspring, the locomotor sensitization 3) The short and long-term plasma endogenous opioids peptides status, and 4) the transcriptomics of brain areas involved in the neural interface between motivation and action.We found that female and male neonates from opioids-treated dams are smaller through the lifespan compared to VEH-exposed counterparts (n=6, p<0.05). Also, adult offspring exposed to MOR in utero displayed sympathetic activation and increased cardiometabolic risk factors along with increased locomotor sensitization to MOR (n=4, p<0.05). In addition, we performed RNAseq analysis of the perivascular nucleolus of the hypothalamus in MOR-exposed offspring, showing increased FOSb expression (n=3 per group). ΔFosB, encoded by the fosB gene, is induced in the brain's reward regions by chronic exposure drugs of abuse and mediates sensitized responses to drug exposure.We also found that rat neonates exposed to opiods showed behavioral NOWS-like symptoms. Importantly, we found that the prenatal exposure to opioids induces a long-term dysregulation of the endogenous opioids peptides (EOP) pro-enkephalin and pro-dynorphin.Both endogenous and exogenous opioids can bind to opioid receptors, which are G-protein coupled receptors (GPCRs) that can dimerize and affect the function of other GPCRs with important neurobehavioral functions.This study provides insights regarding how opioids could affect the programming of the mechanisms regulating blood pressure control later in life. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
MLN alters SERCA1a and SERCA2a kinetics but does not alter the apparent coupling ratio
Authors: Juracic Emma Sara , Brahmbhatt Aditya , Tupling A. Russell 
Published: May 2023

The sarco(endo)plasmic reticulum Ca2+-ATPases (SERCAs) are responsible for inducing muscle relaxation and are integral to the maintenance of intracellular calcium (Ca2+) homeostasis. As such, their activity is modulated by multiple regulatory proteins. Myoregulin (MLN) is a newly discovered protein inhibitor of SERCAs that physically interacts with the pump to regulate Ca2+-handling in muscle. While MLN has emerged as a key regulator of Ca2+ homeostasis and muscle contractility, it is unknown whether MLN can uncouple Ca2+ transport from ATP hydrolysis by SERCAs and in doing so, alter SERCAs’ apparent coupling ratio. To that end, HEK-293 cells were co-transfected with cDNA encoding SERCA1a alone or with SERCA1a and MLN and SERCA2a alone or with SERCA2a and MLN. Both Ca2+ uptake and Ca2+-ATPase activity were measured on crude cell homogenate prepared from the transfected cells. Ca2+-dependent SERCA activity was assessed over Ca2+ concentrations ranging from pCa 6.85 to 4.80 in presence and absence of the Ca2+ ionophore A23187 using a spectrophotometric plate reader assay. SERCA-mediated Ca2+ uptake was measured in the presence and absence of the precipitating anion, oxalate, using the fluorescent dye Indo-1 and a fluorometer. SERCA coupling ratio was calculated by dividing Ca2+ uptake by Ca2+-ATPase rates across different pCa values. In both the presence (p < 0.05) and absence of ionophore (p < 0.05), MLN significantly depressed the maximal rate of ATP consumption (VMAX) and SERCA’s Ca2+ affinity, with this effect being more pronounced in the presence of ionophore. Similarly, MLN significantly reduced SERCA Ca2+ uptake in both conditions, with a greater effect in the presence of oxalate. The ability of MLN to reduce VMAX and impede Ca2+ uptake was greater for SERCA1a compared to SERCA2a. These results indicate that MLN does not affect the Ca2+/ATP coupling ratio of SERCA1a and SERCA2a pumps at maximal (presence of ionophore/oxalate) and physiological (absence of ionophore/oxalate) conditions. NSERC This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Bile acids altered by high fat feeding and oligofructose supplementation impact metabolic homeostasis
Authors: Meyer Rachel , Plett Rannon , Weninger Savanna , Wachsmuth Hallie , Duca Frank 
Published: May 2023

The gut microbiota is recognized as a contributor to host metabolic homeostasis, as obesity and diabetes are associated with unique gut microbiome signatures. Further, strategies aimed at beneficially shifting the gut microbiome, such as prebiotic treatment, show promise in ameliorating metabolic disease; however, the mechanisms are not yet understood. A potential contribution of the gut microbiota to host metabolism is via modification of host and diet-derived metabolites. For example, bile acids, crucial to lipid digestion and absorption, are modified by gut microbes in the small intestine. Bile acids have also been identified as signaling molecules in the intestine, liver, adipose, and nervous system, playing a role in metabolic homeostasis. Despite their importance, few studies have examined the impact of bile acid signaling on mediating the metabolic benefits of prebiotics. We hypothesized that bile acid homeostasis is altered with dietary factors that shift the gut microbiome, including high fat diet (HFD) feeding and supplementation with the prebiotic fiber oligofructose (OFS), and that taurocholic acid (TCA) and glycodeoxycholic acid (GDCA) that are increased with HFD feeding and OFS supplementation, respectively, would recapitulate the metabolic effects of these diets. Therefore, we conducted an untargeted metabolomics analysis on small intestinal contents of rats fed chow, HFD, or HFD supplemented with OFS, identifying bile acid metabolism as a primary contributor to the observed between-group differences. We then quantified bile acid levels in the small intestine, portal vein, and liver of rats fed chow, HFD, or HFD supplemented with OFS, and identified several bile acids of interest that are altered with HFD or OFS supplementation, including TCA and GDCA. Because these bile acids activate the bile acid receptors, FXR and TGR5, that known to impact metabolic homeostasis, we treated HFD-fed mice with 50 mg/kg GDCA or TCA daily for 12 weeks and measured body weight, adiposity, glucose tolerance, food intake, and indirect calorimetry. We found that GDCA treatment improved glucose tolerance, decreased body weight and adiposity, and increased energy expenditure compared to controls. Further, TCA worsened glucose tolerance independent of body weight. The results of this study implicate GDCA and TCA in the beneficial and detrimental effects of OFS and HFD-feeding, respectively; however, more research is necessary to determine the mechanism underpinning the metabolic effects of GDCA and TCA treatment. This work was supported by the NIH R01DK1804. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Respiratory instability in neonatal rats after exposure to gestational intermittent hypoxia
Authors: Sprenger Ryan , Marino Nicholas , Johnson Stephen , Baker Tracy , Watters Jyoti 
Published: May 2023

Sleep disordered breathing (SDB) during pregnancy is a growing concern because it causes adverse outcomes for infant offspring. Recently, our lab developed a rat model of SDB during pregnancy in which pregnant dams were exposed to gestational intermittent hypoxia (GIH; 15 episodes/hr of 10.5% oxygen for 8h daily from days 10-20 of pregnancy). Control dams were exposed in parallel to gestational intermittent normoxia (GNX). We found that male (but not female) GIH offspring had increased apneas in adulthood compared to their GNX control counterparts. Thus, in this study, we tested the hypothesis that GIH would increase respiratory instability (i.e. increase the number of apneas) in neonatal male, not female, offspring. Plethysmography was performed on GIH and GNX male and female pups at postnatal (P) days 0, 10, and 15, and the number of apneas was quantified. An apnea was defined as two missed breaths (2x the average period between breaths). Preliminary data suggest that apnea frequency was sexually dimorphic and changed both with development and treatment. At P10 and P15 male GIH offspring had more apneas compared to GNX offspring (p<0.05). However, at P0, male GIH offspring (16 ± 2.4 apneas/10m) had surprisingly fewer apneas compared to GNX offspring (28 ± 1.9 apneas/10m). In female offspring in contrast, there was no difference in apnea frequency between GIH and GNX treatment at P0, but at P10 and P15 respectively, female GIH offspring (17 ± 2.8 and 9 ± 2.3 apneas/10m) had more apneas compared to their GNX counterparts (8 ± 1.2 and 2.9 ± 0.57 apneas/10m). Apnea frequency decreased with increasing age in both sexes, regardless of GIH and GNX treatment. Together, as expected, these results suggest that the increased apnea phenotype observed in the adult GIH male emerges as early as postnatal day 10. Unexpected however is the observation that female neonatal GIH offspring also experience respiratory instability in during the postnatal period despite its absence in adult female GIH offspring. Studies are currently underway to better understand the mechanisms underlying these sex differences in respiratory neural control, and the protective responses utilized by adult female GIH offspring to mitigate these respiratory deficits induced by intermittent hypoxia exposure in utero. R01 HL142752 This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.