bluuebunny/crossref_metadata_2025_split huggingface dataset:
C‐peptide and insulin at physiological ratios rescue low O2‐induced ATP release from erythrocytes (RBCs) of humans with type 2 diabetes: implications for the prevention of microvascular complications (1108.4)
Authors: Richards Jennifer , Yosten Gina , Kolar Grant , Bowles Elizabeth , Stephenson Alan , Ellsworth Mary , Sprague Randy 
Published: June 2021

RBCs participate in the matching of O₂ delivery with tissue need via release of O₂ and ATP when these cells are exposed to low pO₂. This response is deficient in DM2 RBCs. We reported that incubation of healthy human (HH) RBCs with insulin at postprandial levels inhibits low O₂‐induced ATP release. However, in vivo, insulin is co‐released with equimolar amounts of C‐peptide, but due to different degradation rates, C‐peptide levels exceed those of insulin. We reported that co‐incubation of these peptides, at physiological ratios and concentrations, prevents the adverse effects of insulin on ATP release from HH RBCs.Here we extend these studies to DM2 RBCs. Using Western analysis, we found that the C‐peptide receptor, GPR146, is present on both HH (n=2) and DM2 (n=5) RBCs and pre‐incubation of HH RBCs with an antibody to an external epitope of GPR146 inhibited C‐peptide‐mediated effects (n=3). We next determined that co‐incubation of DM2 RBCs with C‐peptide and insulin at physiological ratios of 1:1 (n=6) or 4:1 (n=8) restored low O₂‐induced ATP release. We also investigated if C‐peptide acts via stimulation of soluble guanylyl cyclase (sGC) activity. The selective sGC inhibitor, ODQ, prevented C‐peptide rescue of low O₂‐induced ATP release in DM2 RBCs. These data suggest a heretofore unrecognized synergism between C‐peptide and insulin that could be important for prevention of the microvascular complications of DM2.Grant Funding Source: Supported by ADA, AHA and NIH
Effect of vitamin D on insulin resistance and tissue inflammation in western and control diet‐fed mice (1108.7)
Authors: Mutt Shivaprakash , Raza Ghulam , Karhu Toni , Keinänen‐Kiukaanniemi Sirkka , Jarvelin Marjo‐Riitta , Herzig Karl‐Heinz 
Published: June 2021

INTRODUCTION: Pandemic Vitamin D (VitD) deficiency is associated with low‐grade chronic inflammation and type 2 diabetes. We hypothesize that vitD could inhibit the inflammatory response and increase the insulin sensitivity.METHODS: 6‐7 week old, male C57BL/6J mice were fed with ± VitD control (CD; 10% fat) or Western (WD; 41% fat) diets for 15 weeks. The VitD deficient (VDD) control and obese mice were further subdivided into two groups. One group was supplemented with vitD (i.p 50ng/mouse/3x/week), the other with saline for 4 weeks. ITT and GTT performed before and after supplementation. Cytokines and tissue inflammatory gene expressions were analyzed.RESULTS: In both VDD CD &amp; WD groups 25‐OH‐vitD levels were reduced compared to controls CD &amp; WD (6.3±1.5; 23.5±0.8 vs 93±5.4; 113.1±9.4, P <0.05), and after supplementation levels were significantly increased (73.3±7.8; 62.1±2.6). Both VDD groups and WD mice had an impaired glucose tolerance compared to the CD mice. After VitD supplementation insulin resistance was reduced and glucose tolerance improved. In addition reduced inflammatory cytokines (IL‐1β: 27.2±8.1 vs 167.6±91.9; RANTES: 79.5±13.3 vs 154±69.7) in CD VDD mice but not in WD‐VDD mice.CONCLUSION: VitD deficiency in both CD and WD fed mice groups induced insulin resistance. Supplementation with VitD in the CD‐VDD improved insulin sensitivity and reduced inflammation, but a similar effect has not been observed in the VDD obese mice.Grant Funding Source: Supported by Academy of Finland, EVO & Finnish Cultural Foundation North Ostrobothnia Regional funds
Bacterial overload enhances duodenal anion secretion via PKA upregulation (1109.15)
Authors: Akiba Yasutada , Strugatsky David , Kaji Izumi , Kaunitz Jonathan 
Published: June 2021

The duodenal mucosa senses luminal bacteria via pattern recognition receptors. Rats, as coprovores, likely populate their foregut with fecal flora, which we hypothesized would enhance anion secretory responses in the duodenum.We measured short‐circuit current (Isc) in Ussing chambered muscle‐stripped mucosa‐submucosa preparations of rat duodenum. Animals were treated with a suspension of homogenized feces (Fecal, 0.1 g/kg, ig) with or without vehicle (Veh) or omeprazole treatment (OPZ, 10 mg/kg, ig) up to 7 days. Duodenal microflora were assessed by culturing duodenal mucosal scrapes on Brucella blood agar or Lactobacillus‐MRS agar plates.Basolateral application of forskolin (FK, 10 µM) increased Isc, which was further increased by the addition of carbachol (CCh, 10 µM). The Isc increase in response to FK and CCh was markedly enhanced in OPZ+Fecal group, compared to Veh group, whereas no differences were noted among Veh+Fecal, OPZ, or Veh groups. Acetazolamide‐sensitive Isc was increased in OPZ+Fecal group, consistent with enhanced HCO3‐ secretion. Expression of protein kinase A (PKA) catalytic domain a and b, and regulatory domain 1a and 2a was significantly increased in Veh+Fecal and OPZ+Fecal groups, while the expression of the cystic fibrosis transmembrane conductance regulator (CFTR), cyclooxygenase (COX)‐2 or tumor necrosis factor (TNF)‐α was not changed. OPZ treatment increased the detection frequency of lactobacillus in the duodenum, and OPZ+Fecal treatment markedly increased lactobacillus colony counts.These results suggest that increased foregut bacterial exposure enhances the anion secretory response to FK in the duodenal mucosa. Since lactobacillus is a major probiotics, upregulation of PKA by probiotics may be a novel pathway to modulate the duodenal mucosal defenses.Grant Funding Source: Supported by VA Merit Review, NIH R01 DK54221
Intestinal epithelial barrier dysfunction protects against pathogen translocation during initial infection (1111.2)
Authors: Edelblum Karen , Singh Gurminder , El Bissati Kamal , McLeod Rima , McCormick Beth , Turner Jerrold 
Published: June 2021

Myosin light chain kinase (MLCK) is essential for pathophysiological epithelial tight junction regulation. Transgenic mice with intestinal epithelial‐specific expression of constitutively‐active MLCK (CA‐MLCK) demonstrate increased permeability and susceptibility to immune‐mediated colitis. However, the effect of barrier dysfunction on enteric infection remains unknown. Within 30 min after mucosal application of DsRed‐labeled Salmonella typhimurium (SL3201 108 CFU), CA‐MLCK mice exhibited a 5‐fold reduction in bacterial translocation compared to wildtype (WT). This protection also occurred 1 h after oral infection with Toxoplasma gondii, in which tachyzoite invasion was reduced in CA‐MLCK mice relative to WT, 1.5±0.6 vs 4.0±1.0 tachyzoite/mm2 villus, respectively. Intraepithelial lymphocytes expressing the γδ T‐cell receptor (γδ IEL) protect against pathogen invasion. Time‐lapse confocal microscopy revealed enhanced γδ IEL surveillance of the villous epithelium in CA‐MLCK mice compared to WT with more rapid migration (4.2±0.1 vs 3.8±0.1μm/min) and reduced localization to the lateral intercellular space (26±1.7 vs 32±3.3%). However, loss of γδ T cells in CA‐MLCK mice did not alter Salmonella transmigration compared to WT. These data indicate that increased γδ IEL epithelial surveillance is not sufficient for the reduced pathogen invasion observed during early infection in CA‐MLCK mice.Grant Funding Source: Supported by NIH DK61931 and DK68271, UChicago DDRCC, Gastro‐intestinal Research Foundation
Stabilisation of epithelial hypoxia‐inducible factor reverses colitis through accelerated restitution (1111.3)
Authors: Goggins Bridie , Minahan Kyra , Kostakis Anna , Shalwitz Robert , Horvat Jay , Keely Simon 
Published: June 2021

During mucosal inflammation, changes in metabolic activity and vascular tissue damage, lead to a reduction in tissue‐oxygen tension (hypoxia), in which healing processes such as re‐epithelialisation occur. Previous work has shown that pharmacological induction of hypoxia‐inducible factor (HIF), a global transcriptional regulator of the hypoxic response, by prolyl hydroxylase inhibitors (PHDi) is protective in murine models of colitis.We have previously identified induction of integrin β1 (ITGB1) during hypoxia. As integrins play critical roles in epithelial wound healing, we hypothesised that pharmacological HIF activation accelerates epithelial restitution through integrin‐dependent pathways. We aimed to investigate the relative induction of integrin β1 heterodimers in epithelial cells by PHDi in vivo and in vitro.Colitis was induced in BalbC mice by rectal instillation of 2.5% TNBS. PHDi (Akebia‐4924, [5mg/Kg]) was administered by oral gavage. Colitis was monitored endoscopically over 7 Days with the Karl Storz coloview endoscope. Biopsies were taken on alternate days with a 20‐french biopsy forceps. Mice were euthanised and tissues and colonic epithelial cells were harvested for mRNA and protein. Scratch assays were performed on Caco‐2 monolayers seeded on 6 well plates.TNBS colitis led to a significant increase in ITGB1 transcript in epithelium, correlating (R2=0.71) disease severity. Mice treated with PHDi demonstrated advanced endoscopic healing compared to vehicle treated animals, concurrent with an earlier induction of ITGB1 and induction of epithelial integrins α3 and α6. Caco‐2 monolayers show accelerated wound healing in scratch assays when treated with 10mM PHDi which was inhibited by antibodies functionally targeting α3β1 and α6β1 integrin homodimers. These data suggest that mucosal HIF activation promotes healing through accelerated induction of epithelial intregrins.Grant Funding Source: National Health and Medical Research Council
Risk factors of diseases gastrointestinal path (1112.1)
Authors: Imashev M. , Dubitsky A. , Fursov A. 
Published: June 2021

Determination of risk factors of diseases of a gastrointestinal path at teenagers.Material. Under supervision there were 224 teenagers from 15 to 18 years, from them 112 teenagers with pathology of a digestive tract and 112 control group. Diagnostics was carried out on the basis of standard criteria.Results. Non‐compliance with a diet took place at 33 (29,5%) teenagers of the main group, at 11 (9,8%) control (3,0 points). The frequent use of hot and smoked dishes was observed at 17,8% teenagers of the main group and at 8% control (2,0 points). Negative emotions are established almost at each 5‐6 teenagers ‐ 17,8% the main group and at 8% control. Bad living conditions are established at 24,1% 1 groups and at 13,4% 2 groups 1,8 points. In 1 group have addictions more than 28%, from them smoke 22,3%, take alcoholic beverages 8,9%. In control group 7,1% and 2,7%, respectively (3,0 and 3,3 points).1 group (low risk) included teenagers at whom total amount of points on risk factors didn't exceed 5; the 2nd group included teenagers with a score from 5 to 10; the 3rd group ‐ high risk ‐ included children with the sum of the gained points 10,0 and more. Within a year medical and improving events at 35 teenagers of group of high risk were held. In group of high risk there were only 19 teenagers as they passed to groups of an average and low risk, and the part of teenagers was struck off the register. The indicator efficiency of improvement made 45,7%.
Nitric oxide synthase inhibition exacerbates liver injury in an obese rat model of NAFLD (1115.3)
Authors: Sheldon Ryan , Padilla Jaume , Jenkins Nathan , Laughlin M. , Rector R. 
Published: June 2021

We tested the hypothesis that chronic disruption of constitutive nitric oxide (NO) production via the non‐selective NO synthase (NOS) inhibitor Nω‐nitro‐L‐arginine methyl ester (L‐NAME) would intensify liver injury in a rat model of obesity, insulin resistance, and nonalcoholic fatty liver disease (NAFLD). Obese Otsuka Long‐Evans Tokushima Fatty (OLETF) and lean Long‐Evans Tokushima Otsuka (LETO) rats (16‐wks) received either control or L‐NAME (65‐70 mg/kg/d) containing drinking water (n=10/group) for 4‐wks. L‐NAME treated OLETF and LETO rats had significantly (p < 0.05) reduced serum NO metabolites (NOX) and food intake. Despite reducing % body fat (‐2.5%; p < 0.05) in OLETF rats, L‐NAME treatment increased hepatic triacylglycerol (TAG) content (+40%; p < 0.05) compared to control OLETF rats. In addition, L‐NAME treatment caused a shift toward M1 macrophage polarity as indicated by elevated CD11c (p<0.05) and IL‐1β (p=0.07) mRNA in the OLETF rats. Markers of total and resident macrophages (CD68 and CD163 mRNA, respectively), fibrosis (TGFβ mRNA), and cell adhesion (MCP‐1 and ICAM mRNA) were unaffected by L‐NAME treatment in both groups. In conclusion, chronic NOS inhibition appears to aggravate liver injury by increasing expression of macrophage activation‐related genes associated with insulin resistance and by accelerating hepatic TAG accumulation in obese, insulin resistant rats.Grant Funding Source: Supported by a Life Sciences Fellowship (RDS), NIH HL‐36088 (MHL), and VHA‐CDA2 BX001299‐02 (RSR)
Secretory and absorptive dysfunctions in ob/ob mouse jejunum are sex‐dependent (1117.1)
Authors: Al‐Nakkash Layla , Leung Lana 
Published: June 2021

Sex‐dependent differences in intestinal absorption and intestinal secretion in the ob/ob leptin‐deficient obese mouse are poorly understood. We used male and female ob/ob and lean mice aged ~14 weeks. The goal of this study was to characterize jejunal function in this clinically relevant mouse model. We measured transepithelial short circuit current (Isc), a measure of chloride secretion, across freshly isolated segments of jejunum from ob/ob and lean mice. Compared to lean controls, basal Isc was significantly decreased by 24 µA/cm2 (P<0.05, n=9) in ob/ob females and significantly decreased by 30 µA/cm2 (P<0.05, n=10‐11) in ob/ob males. Reduced basal Isc in ob/ob males is partly attributed to a significant 2‐fold decrease in CFTR expression. The contribution of key transporters involved in promoting chloride secretion are currently under evaluation. While crypt depth was unchanged in ob/ob males, the number of goblet cells/crypt was 2‐fold greater and this may also account for, in part, the reduced basal Isc in ob/ob males. Villi length and total small intestinal length were both significantly increased in ob/ob female and male mice, suggesting a greater surface area for absorptive function. These structural changes likely contribute towards the obese phenotype. Expression of the fructose transporter protein, GLUT5 (normalized to GAPDH), was significantly increased 1.9‐fold (P<0.05, n=12‐15) in male ob/ob mice compared to leans, which likely contributes to the diabetic/obese phenotype of ob/ob males. Expression of the glucose co‐transporter protein, SGLT1 (normalized to GAPDH), was significantly decreased 30% (P<0.05, n=8‐9) in female ob/ob mice compared to leans. These data suggest that basal jejunal Isc in ob/ob mice is deficient, and may contribute towards slower transit times in the gastrointestinal tract in the ob/ob mice. Interestingly, while both female and male ob/ob mice are obese, the specific secretory and absorptive dysfunctions are not comparable in both sexes.Grant Funding Source: Supported by NIH R15 DK071625‐01A2
Cultured milk bacterial isolates survive well in digestive tract conditions and could be potential probiotic candidates (1118.4)
Authors: Boyiri B , Kotala J , Lampson B , Onyango E 
Published: June 2021

Probiotics are viable non‐pathogenic microbes that positively affect host health. Our goal is to understand the influence of probiotics on the intestinal barrier function. In this study bacterial isolates from traditional cultured milk from Kenya were screened for their ability to withstand digestive tract conditions and thence their potential as probiotic candidates. Bacteria in the sample were isolated and identified by single colony PCR sequence match analysis of the 16s rRNA gene. Identified isolates included Lactobacillus spp (6 isolates), Acetobacter spp (2 isolates), Bacillus spp (2 isolates) and 1 Staphylococcus sp. Isolates were examined for acid stability over a 3‐h period, acid and/or bile stability over a 9‐h period, antimicrobial activity, and antibiotic sensitivity to ampicillin, bacitracin, chloramphenicol, erythromycin, kanamycin, penicillin, streptomycin and tetracycline. Two Lactobacillus spp isolates were stable in acid and/or bile, showed antimicrobial activity against K. pneumonia and P. aeruginosa and were sensitive to erythromycin, kanamycin, streptomycin and tetracycline. The remaining 4 Lactobacillus isolates were unstable in acid and/or bile, showed antimicrobial activity against K. pneumonia and P. aeruginosa and were sensitive to all tested antibiotics. The Bacillus spp isolates were acid and/or bile stable, showed no antimicrobial activity, and were susceptible to all the tested antibiotics. The Staphylococcus sp and 2 Acetobacter spp were acid and bile unstable and did not have antimicrobial activity and were sensitive to the antibiotics tested. In conclusion, the Lactobacillus isolates that showed stability in acid/bile conditions, demonstrated antimicrobial activity and were sensitive to antibiotics could be potential probiotic candidates.Grant Funding Source: East Tennessee State University Research Development Committee Small Grants Program
CagA+ Helicobacter pylori induces proliferation and expression of Sonic hedgehog in fundic organoid model (1119.4)
Authors: Schumacher Michael , Aihara Eitaro , Feng Rui , Zavros Yana 
Published: June 2021

Helicobacter pylori (H. pylori) infection leads to aberrant cell proliferation and dysregulated Hedgehog signaling in the stomach. The organoid model is an emerging in vitro system whereby isolated gastric glands are cultured into three‐dimensional spheres that express cells present in the original tissue. This model allows for the study of epithelial responses to infection in a system independent of recruited inflammatory cells. We hypothesize that H. pylori induces cell proliferation and acute expression of Sonic hedgehog (Shh) in the fundic organoid model in the absence of recruited circulating host factors. Fundic organoids were generated from dissociated gastric glands of control or transgenic mice lacking Shh expression in the parietal cell (PC‐ShhKO). Expression of mature cell lineage markers was confirmed by RT‐PCR and immunofluorescence staining. Colonization and epithelial attachment was confirmed by silver stain, live‐imaging and quantitative re‐culture. Organoids injected with cagA+ but not cagA‐deficient H. pylori had significantly increased cell proliferation as measured by EdU staining. Proliferation was associated with c‐Met phosphorylation as determined by western blot. H. pylori induced Shh at 1 day post‐injection in control organoids but not those generated from PC‐ShhKO mice, and this effect was inhibited by NFκB inhibition. Inhibition of CXCL1/2 signaling, a putative mediator of Shh induction, did not block H. pylori‐induced Shh expression. This report demonstrates a uniquely beneficial in vitro approach to study the host response to H. pylori interactions with the gastric epithelium, independent of recruited hematopoietic factors. Fundic organoids recapitulate the in vivo responses to H. pylori of cell proliferation and Shh induction and provide mechanistic insight demonstrating 1) cagA dependence and c‐Met phosphorylation association with H. pylori‐induced proliferation and 2) NFκB requirement for H. pylori‐induced gastric Shh expression.Grant Funding Source: Supported by NIH 1R01DK083402‐01 (Y. Zavros) and AGA: Funderburg Research Award (Y. Zavros)
Improving post‐traumatic stress disorder biomarkers through matching patients and controls on multiple criteria (1120.10)
Authors: Donohue Duncan , Miller Stacy‐Ann , Gupta Ellen , Yang Ruoting , Guatam Aarti , Hammamieh Rasha , Jett‐Tilton Marti 
Published: June 2021

Molecular typing of psychological disease depends on a variety of complicated environmental and phenotypic factors. The quality of results critically depends on the cohort selection criteria, including those criteria used to match case and controls. Our PTSD biomarker study initially matched patient groups on age and self‐reported race. Further analysis has shown striking differences in the types and amounts of medications used by PTSD patients vs. controls. Here, we present an approach to simultaneously match patient groups across age, ancestry, and medication use applying random sampling methods. We compare receiver operating characteristics to quantify the degree to which matching on these three variables generates blood mRNA expression biomarkers which better discriminate patients from controls. Work is in progress to fine‐tune matching by incorporating the relative effect size of these three measures.Disclaimer: Research was conducted in compliance with the Animal Welfare Act and all other Federal requirements. The views expressed are those of the authors and do not constitute endorsement by the U.S. Army. Human blood sample use protocol was conducted at USACEHR under HRPO Log number A‐15459.f.
Base‐resolution maps of 5‐methylcytosine and 5‐hydroxymethylcytosine in Dahl salt‐sensitive rats (1120.8)
Authors: Liu Yong , Liu Pengyuan , Yang Chun , Cowley Allen , Liang Mingyu 
Published: June 2021

Analysis of 5‐hydroxymethylcytosine (5hmC) at single‐base resolution has been largely limited to studies of stem cells or developmental stages. Given the potential importance of epigenetic events in hypertension, we have analyzed 5hmC and 5‐methylcytosine (5mC) at single‐base resolution in the renal outer medulla of the Dahl salt‐sensitive SS rat and examined the effect of disease‐relevant genetic or environmental alterations on 5hmC and 5mC patterns. Of CpG sites that fell within CpG islands, 11% and 1% contained significant 5mC and 5hmC, respectively. 5mC levels were substantially higher for genes with lower mRNA abundance and showed a prominent nadir around the transcription start site. In contrast, 5hmC levels were lower in genes with lower expression. Substitution of a 12.9 Mbp region of chromosome 13, which attenuates the hypertensive and renal injury phenotypes in SS rats, or exposure to a high‐salt diet, which induces the disease phenotypes, was associated with differential 5mC or 5hmC in several hundred CpG islands. Nearly 80% of the CpG islands that were differentially methylated in response to salt and associated with differential mRNA abundance were intragenic CpG islands. The substituted genomic segment had significant cis effects on mRNA abundance but not DNA methylation. The study established base resolution maps of 5mC and 5hmC in an in vivo model of disease and revealed several characteristics important for understanding the contribution of 5mC and 5hmC to hypertension and other complex diseases.Grant Funding Source: Supported by the NIH.