bluuebunny/crossref_metadata_2025_split huggingface dataset:
Nox1 Inhibition Attenuates Non‐Alcoholic Fatty Liver Disease and De Novo Lipogenesis
Authors: Larion Sebastian , Thompson Jennifer , Butcher Josh , Haigh Stephen , Mintz James , Khurana Sandeep , Fulton David , Stepp David 
Published: November 2023

ObjectiveInvestigate the role of NADPH oxidase‐1 (Nox1) in the development of non‐alcoholic fatty liver disease (NAFLD).MethodsNox1 mRNA expression was investigated in human liver repository data [GDS4881] and confirmed in hepatic tissue of db/db mice. Deletion studies investigating the role of Nox1 in the development of NAFLD were conducted by generating constitutive Nox1 knockout mice bred on a db/db background. Metabolic syndrome was assessed using biochemical and histological indices. Hepatic mRNA expression was assessed using microarray (Affymetrix) with genes of interest confirmed using RT‐PCR. Lipid species accumulation was assessed by GC‐MS with ROS production evaluated by 8‐OHdG and nitrotyrosine immunofluorescence.ResultsNox1 expression was significantly increased in human liver biopsies from patients with simple steatosis over healthy or obese patients without NAFLD. Increased expression of Nox1 and associated subunits was also evident in liver from db/db mice vs. lean (all p<0.05). Mouse studies revealed that obese mice had significantly greater food intake, anthropomorphic (body weight, visceral adiposity, lean muscle) and biochemical evidence of metabolic syndrome including dyslipidemia (serum triglycerides, cholesterol, free fatty acids), and insulin resistance (insulin, fasting blood glucose, HgbA1c, glucose tolerance test) than lean controls, with no difference vs obese mice with Nox1 deletion. Liver mass was increased in obese vs lean controls, but significantly attenuated (−36%; p<0.05) in obese mice with Nox1 deletion. Obese mice with Nox1 deletion also had a striking reduction in macroscopic nodularity and pale yellow coloring, with reduced ORO staining and triglyceride content (−69%; p<0.05) than controls. NMR revealed significantly increased fat accumulation in obese mice vs lean controls, which was reduced (−23%; p<0.05) in obese mice with Nox1 deletion. Similarly, SFA species and ROS production were increased in obese mice, but significantly attenuated in obese mice with Nox1 deletion (all p<0.05). Measurement of mRNA expression via RT‐PCR revealed increased expression of hepatic SREBF1 and its downstream effector SCD1, the rate limiting enzyme of hepatic MUFA production, in obese mice vs controls, which were both significantly decreased in obese mice with Nox1 deletion. Pathway analysis disclosed SREBF1 as a major regulator in the highest scoring network: Lipid metabolism (score 35).ConclusionNox1 inhibition attenuates hepatic steatosis in a genetically obese mouse model, likely through alterations in de novo lipogenesis. Nox1 signaling may prove to be a viable therapeutic target in NAFLD.Support or Funding InformationNIHHL124691
Pharmacologic activation of the sarco‐endoplasmic reticulum ATPase (SERCA) reduces palmitate‐mediated endoplasmic reticulum stress in liver cells
Authors: Estrada Andrea L , Stewart Claire M , Kim Paul Y , Wang Dong , Wei Yuren , Pagliassotti Michael 
Published: November 2023

PurposeElevated fatty acids, in particular saturated fatty acids, have been shown to induce endoplasmic reticulum (ER) stress in numerous cell types including hepatocytes. However, the specific mechanisms that link saturated fatty acids to ER stress are poorly understood. Calcium is a critical cofactor for chaperone‐mediated protein folding in the ER. Previous studies have demonstrated that palmitate, a saturated fatty acid, reduces ER lumenal calcium stores, which are maintained by the activities of three ER‐bound proteins: the inositol triphosphate receptor, the ryanodine receptor, and SERCA. In the present study, we examined whether palmitate‐induced ER stress could be mitigated via activation of SERCA.MethodsH4IIE liver cells (n=3) were treated with control media or control media supplemented with palmitate (250μM), a saturated fatty acid that induces ER stress, oleate (250μM), an unsaturated fatty acid that does not induce ER stress, or a combination of both palmitate and oleate that does not induce ER stress. Treatments were administered in the presence or absence of CDN1163 (10μM), a SERCA pump activator, for 6 hours.ResultsOnly palmitate treated cells were characterized by increased ER stress. Co‐incubation with CDN1163 reduced palmitate‐mediated ER stress. These data suggest that palmitate‐mediated ER stress involves a reduction in SERCA activity.Support or Funding InformationNIH: DIK072017
Fecal Supernatants from Active Ulcerative Colitis Patients Impair Colonic Epithelial Barrier Integrity and Activate Pelvic Mucosal Extrinsic Afferent Nerves
Authors: Wardill Hannah R , Dmochowska Nicole , Campaniello Melissa A , Mavrangelos Chris , Bowen Joanne M , Andrews Jane M , Costello Sam P , Hughes Patrick A 
Published: November 2023

IntroductionUlcerative Colitis is characterised by colonic mucosal inflammation. Microbial dysbiosis is well documented in active UC, but little is understood regarding the physiological implications this has on epithelial barrier integrity or colonic extrinsic afferent signalling. We aimed to determine the effects of fecal supernatants from active UC on epithelial barrier and colo‐rectal mucosal extrinsic pelvic afferent nerves.MethodsExcreted fecal samples from 10 patients with active UC (endoscopic MAYO subscore ≥2) were mixed with Ringers (Ussing) or Krebs solution (electrophysiology) at 1.5gm / 3 ml, centrifuged, and the supernatant filtered and pooled. Pooled fecal supernatants (FSN) were run over a 30kDa size exclusion column. Neat, high and low molecular weight FSN were applied to CACO‐2 monolayers or healthy mouse colon tissue in Ussing chamber experiments over a 2 hour period. Extracellular electrophysiological responses from pelvic colonic extrinsic afferent nerves were recorded from healthy mice. Mucosal afferents were characterised as responding to fine mucosal stroking with 10 mg Von Frey Hair. Neat FSN was applied to mucosal afferents for 5 min and mechanical sensitivity to graded mucosal stroking with Von Frey Hair re‐assessed.ResultsNeat UC FSN potently decreased the resistance of CACO‐2 monolayers and mouse colon over a 2 hour period compared to vehicle. Low molecular weight FSN also decreased the resistance of mouse colon, yet increased the resistance of CACO‐2 monolayers. High molecular weight FSN had no effect on either mouse tissue or CACO‐2 monolayers resistance. Neat UC FSN directly activated mucosal colonic afferent endings immediately following addition, but had no effect on mechanical sensitivity to mucosal stroking.DiscussionMediators present in faecal samples impair the resistance of the epithelial barrier and directly activate mucosal afferent nerve endings. The <30kDa fraction had the most potent activity on epithelial barrier. Further characterisation of the active mediator(s) and mechanisms has the potential to lead to new targets for the treatment of barrier dysfunction and nerve activation in active UC.Support or Funding InformationSupported by NHMRC Australia
Control of lupus nephritis by the gut microbiota
Authors: Mu Qinghui , Zhang Husen , Luo Xin 
Published: November 2023

Dysbiosis of gut microbiota promotes autoimmune disorders that damage extraintestinal organs. Here we report the role of gut microbiota in the pathogenesis of renal dysfunction in systemic lupus erythematosus. In a classical model of lupus nephritis, MRL/lpr, we found marked depletion of Lactobacilli in the gut microbiota. Increasing Lactobacilli in the gut improved renal function of these mice. Further studies revealed that MRL/lpr mice possessed a “leaky” gut, which was reversed by Lactobacilli. Lactobacillus treatment decreased IL‐6 and increased IL‐10 in the gut, thereby providing an anti‐inflammatory environment. It also increased IL‐10 and decreased IgG2a in the circulation, the latter a major immune deposit in the kidney of MRL/lpr mice. Inside the kidney, Treg‐Th17 balance was skewed towards Treg with Lactobacillus treatment. These effects of Lactobacilli were present in female and castrated male mice, but not unaltered males, suggesting that gut microbiota controls lupus nephritis in a sex hormone‐dependent manner.Support or Funding InformationThis work was supported by Xin Luo's start‐up funds provided by Virginia Tech College of Veterinary Medicine.
Lactobacillus casei Cell‐Surface Components and Their Impact on Immunomodulation and Cecal Microbiata Composition
Authors: Aktas Busra , De Wolfe Travis J. , Safdar Nasia , Darien Benjamin J. , Steele James L. 
Published: November 2023

Lactobacillus casei are commonly utilized as a probiotic in a wide‐range of fermented and unfermented dairy products. Previously it has been shown that L. casei have extensive strain‐to‐strain genetic variation and that significant strain‐to‐strain differences are present in their ability to alter the gut microbiota and modulate the host innate immune system. The objective of this study is to evaluate L. casei for potential mechanisms resulting in the observed probiotic function. We utilized comparative in silico analysis to investigate genes putatively encoding surface components, including: proteins exposed to the surface; pili; wall teichoic acids (WTA); and lipoteichoic acids (LTA). We also evaluated in vitro the ability of six L. casei strains to modulate cytokine production (IL‐10 and IL12) by intestinal epithelial cells (IEC‐6). Additionally, we investigated the contribution of cell surface components and small molecules produced by L. casei to their probiotic function by examining the effect of live, UV‐killed, and supernatants from L. casei strains on cytokine production by IEC‐6. To evaluate the ability of UV‐killed L. casei to alter the gut microbiota and modify the immune system in vivo, we fed mice L. casei 32G at dose of 108 CFU/day/mouse for seven days and sacrificed 3.5h after the last administration. The cecal content and the ileum tissue were collected for microbiota analysis and immune profiling, respectively. The in silico results demonstrated that there is an extensive genetic diversity within L. casei cell surface components. The in vitro cytokine assay revealed that the strains modulate cytokine expression in a strain specific manner. The cell surface component of all the strains examined reduced the IL‐10 expression while small molecules were determined to modulate IL‐12 expression in some strains, indicating that both cell surface components and small molecules participate in immunomodulation. The in vivo results demonstrated that only live 32G significantly altered the overall microbiota. The effect of live and UV‐killed 32G on the expression of genes involved in innate immune system was gene specific. In conclusion, significant variation occurs between L. casei strains in the mechanism by which they signal with the host innate immune system and hence their mechanisms of probiotic action.Support or Funding InformationThis work was funded by the DuPont Inc (Grant no. PRJ19MQ) and by the WARF (Grant no. PRJ66JZ)
Feeding obese diabetic mice a genistein diet induces thermogenic and metabolic change
Authors: Rockwood Schuyler , Sandoval‐Skeet Noemy , Broderick Thomas L , Al‐Nakkash Layla 
Published: November 2023

Obesity is associated with elevated plasma levels of glucocorticoids, which is known to affect food intake and energy expenditure. Further, tissue specific glucocorticoid metabolism is altered in obesity, and may increase insulin resistance and cardiometabolic risk. We have previously demonstrated that obese (ob/ob) female mice fed a genistein‐based diet resulted in a significant decrease in body weight, fat pad weight, and improved cardiovascular risk markers. The goal of this study was to examine if those changes were sex‐dependent. In order to test our hypothesis, lean and ob/ob male mice were fed either a genistein‐rich diet (600 mg genistein/kg) or a genistein‐free diet, over a 4‐week period. During the 4‐week study, measures of daily diet intake, body weights, and body temperatures were taken. At the conclusion of the diet study, mice were euthanized and organs were removed, weighed and immediately frozen for further analysis. Ob/ob mice weighed significantly more (51.50±0.72 g, n=6, P<0.05) than leans (30.38±2.08 g, n=5) and this was associated with significant increases in liver and total fat pad weight. While there was no genistein effect on weight loss, genistein‐fed ob/ob mice showed a significant increase in overall food consumption in weeks 3 and 4 compared to control ob/ob mice. Interestingly, body temperatures of ob/ob mice after 4‐weeks on the diets were significantly less (30.80±0.33 °C, n=6, P<0.05) than those of lean controls (33.30±0.28 °C, n=5), and genistein diet reversed this (32.66±0.36 °C, n=5, P<0.05), indicating a thermogenic effect. Western blot analysis of liver samples revealed that 11βHSD‐I, which converts inactive corticosterone into active corticosterone, was significantly decreased in ob/ob mice (0.34±0.05, n=6, P<0.05) compared to leans (0.79±0.18, n=4), and there was no genistein‐effect. Protein analysis of kidney 11βHSD‐II, which converts corticosterone into in its inactivated form, was increased with genistein (0.93±0.03, n=5) compared to ob/ob controls (0.68±0.06, n=5, P<0.05). Based on these results, we predict a corresponding improvement in serum corticosterone levels as well as a potential rescue effect from excessive glucocorticoid production by the kidney in genistein‐fed ob/ob mice.Support or Funding InformationFinancial support: MWU intramural funds (LA and TLB), DAREF (LA) and Department of Biomedical Sciences (SR, NS‐S)
G protein‐coupled receptor 30 (GPR30), a novel estrogen receptor, plays an independent role in the pathogenesis of estrogen‐induced cholesterol gallstones in female mice
Authors: Wang David Q , de Bari Ornella , Wang Helen H 
Published: November 2023

BackgroundEstrogen is an important risk factor for gallstone formation because gallstone prevalence is significantly higher in women than in men. The classical estrogen receptor α (ERα), but not ERβ, in the liver plays a critical role in estrogen‐induced gallstones in female mice. Understanding the molecular mechanisms underlying the lithogenic effect of estrogen on gallstone formation became more complicated with the identification of GPR30 as a new gallstone gene Lith18. Our aim is to explore whether GPR30 produces an independent lithogenic action, or works in conjunction with ERα, to promote estrogen‐induced gallstone formation in female mice.MethodsThe biliary and gallstone phenotypes were studied by physical‐chemical methods in ovariectomized (OVX) female GPR30 (−/−), ERα (−/−), and WT mice injected daily with the potent GPR30‐selective agonist G‐1 at 0 or 200 ng/day and fed a lithogenic diet for 8 wk. Gene expression was studied by qPCR and Western blotting.ResultsActivation of GPR30 by G‐1 enhanced cholelithogenesis by suppressing expression of cholesterol 7α‐hydroxylase (CYP7A1), the rate‐limiting enzyme for the classical pathway of bile salt synthesis. This abnormality led to biliary cholesterol hypersecretion associated with reduced bile salt secretion, causing cholesterol‐supersaturated gallbladder bile and accelerating cholesterol crystallization. The prevalence of gallstones was 80% in OVX ERα (−/−) and WT mice treated with G‐1 compared to 10% in OVX WT mice receiving no G‐1. Moreover, G‐1 treatment did not influence gallstone formation in OVX GPR30 (−/−) mice. G‐1 treatment led to a significant increase in mRNA levels and protein concentrations of the epidermal growth factor receptor (EGFR) and a significant decrease in expression of liver receptor homolog‐1 (LRH‐1), coupled with reduced expression of CYP7A1 in OVX ERα (−/−) and WT, but not GPR30 (−/−) mice. In vitro studies using cultured primary hepatocytes isolated from ERα (−/−) and WT mice showed that when EGFR was inhibited by AG1478, a highly potent EGFR kinase inhibitor, expression of CYP7A1 was unchanged even though GPR30 was activated by G‐1.ConclusionsGPR30 plays an independent role in the pathogenesis of estrogen‐induced gallstones in female mice. EGFR is required for GPR30 to exert a non‐transcriptional regulatory role in suppressing hepatic expression of CYP7A1 and the classical pathway of bile salt synthesis, leading to the availability of excess amounts of cholesterol for hepatic hypersecretion and the formation of supersaturated bile. Both GPR30 and ERα are potential targets for preventing cholesterol gallstones, particularly in women and patients who expose to high levels of estrogen.Support or Funding InformationThis work was supported in part by research grants DK101793 and DK106249, both from the National Institutes of Health (US Public Health Service).
Inhibition of p53 with its specific inhibitor pifithrin‐α prevents hepatic oxidative stress and inflammation, probably via up‐regulating Sirt1/Nrf2 pathway
Authors: Qian Lingbo , Gu Junlian , Ji Honglei , Cai Lu 
Published: November 2023

Background and objectiveUp‐regulation of the tumor suppressor p53 has been reported to mediate the diabetic hepatic injury, including oxidative stress and inflammation. However, its underlying mechanisms remain unclear. Nuclear factor erythroid‐derived 2‐like 2 (Nrf2) is a potent factor against diabetic oxidative stress by inducing the expression of its downstream antioxidants. It is revealed that Nrf2 activates its target genes through an antioxidant‐response element (ARE) when Sirtuin 1 (Sirt1) triggers the release of Nrf2 from Nrf2/Keap1 complex. Furthermore, inverse relationship between Sirt1 signaling and p53 protein abundance has been found in high glucose‐exposed hepatic cells. The aim of present study was to explore whether the hepatic protection from diabetes by p53 inhibition with pifithrin‐α is associated with up‐regulation of Sirt1/Nrf2 pathway in type 1 diabetic mice.Methods and ResultsFVB mice were treated with five daily injections of 50 mg/kg streptozotocin to induce type 1 diabetes. Both hyperglycemic and age‐matched control mice were treated with or without pifithrin‐α five times a week for 2 months and then kept without treatment until 3 months. At three months, diabetes significantly increased serum ALT activity, hepatic 3‐NT, 4‐HNE, IL‐6 and TNF‐α expressions, which was significantly prevented by inhibition of p53 expression with pifithrin‐α in diabetic mice. Mechanistically, the inhibition of p53 reversed diabetes‐increased hepatic expression of Keap1 and p47 phox and decreased Sirt1 expression and Nrf2 expression and function.ConclusionsThese results demonstrate that the inhibition of p53‐mediated down‐regulation of Sirt1/Nrf2 pathway by administration of diabetic mice with p53 specific inhibitor can effectively prevent the development of hepatic oxidative stress and inflammation. Therefore, intervention targeting to p53 may be a potential therapy for diabetic hepatic injury.Support or Funding InformationThis work was supported in part by the Scholarship awarded by China Scholarship Council (No. 201508330753), and the grant awarded by National Natural Science Foundation of China (No. 81470495).
Presence and Variations of the Palmaris Longus in a Small Population
Authors: Monin Lydia , Hurley Mary , Olivieri Marion P. 
Published: November 2023

Dr. Frank Jobe first performed the wezl‐known “Tommy John Surgery” on a Major League baseball player in 1974. This surgical procedure involves the reconstruction of the ulnar collateral ligament (UCL) of the elbow. The Palmaris longus, a tendon located in the forearm, has been successfully used as a replacement tendon for the reconstructive surgery. Part of the preoperative plan of the reconstruction includes examination of the Palmaris longus presence, whether bi‐lateral, unilateral or absent; variations may occur that lead to a change in the surgical steps taken during the procedure. More knowledge on the different possible variations of the tendon involved with this surgery will allow medical professionals to have a better understanding of the orientation of the muscle/tendon involved in Tommy John reconstruction. The Palmaris longus normally arises from the medial epicondyle of the humerus, and inserts to the palmar aponeurosis. It exhibits several variations including its overall presence. Thompson did a clinical study on 300 subjects observing the presence of the muscle/tendon in 2001 that concluded forty‐nine subjects had unilateral absence, and bilateral absence in 26 subjects. In those subjects with unilateral absence, the side more commonly affected was the right side. In this study, 12 cadavers were examined for the general presence of the Palmaris longus. Out of the 12 cadavers examined only 3 presented with an absence of the tendon, of those three cadavers all expressed a unilateral absence of the tendon in the left arm. This study indicates that unilateral absence was considerably more prevalent in the left arm than what was originally reported reported by Thompson in 2001, continuing studies will examine additional cadavers and assess significance.Support or Funding InformationSupported and Funded by the Department of Biology and Mathematics of D'Youville College, Buffalo NY
Anatomical Variation of Arterial Supply to the Prostate in a Cadaver Population
Authors: Danovskis Daniel , Gervasi Megan E , Hurley Mary , Olivieri Marion P 
Published: November 2023

For men in the United States, aside from the skin, the prostate is the most common cancer site. Roughly 1 of 7 males will be diagnosed with prostate cancer. This does not include the numerous undocumented signs of prostate cancer identified postmortem. In 2015, it was estimated that in 2016 there would be180,890 new cases of prostate cancer and the disease would cause 26,120 deaths. The vasculature of the prostate differs among individuals, and is not completely understood. Typically, the prostate is supplied by arteries arising from the internal pudendal artery, however there are many undefined variations. The position of the prostate makes it difficult to study during standard gross anatomy course dissection. This study involved development of a protocol for isolation and examination of cadaveric prostates.It was found that observation of the prostate gland was enhanced by isolating the pelvic region. This involved careful removal of the sigmoid colon followed by cuts through both thighs including the femur just inferior to the inguinal region ligament. The vertebral column was detached from the sacrum by cutting through the intervertebral disc between L5 and the sacrum. Once the pelvis was isolated it could be positioned to allow access to the internal iliac artery. The branches of internal iliac were then identified. This was then followed by a midsaggital cut through the entire pelvis enabling greater access to the prostate gland. This also allowed for confirmation of the targets of the branches of the internal iliac and for further dissection revealing how those branches continued into the prostate. Future studies will involve examination of the arterial supply of the prostates of 12 male cadavers. Further understanding of the prostate arterial supply may lead to innovation regarding treatment of prostate cancer.Support or Funding InformationWork supported by the Department of Biology and Mathematics, D'Youville College.