bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibition of Peroxiredoxin 6 PLA2 Activity Decreases Oxidative Stress and the Severity of Acute Lung Injury in the Mouse Cecal Ligation and Puncture Model
Authors: Chatterjee Shampa , Dodia Chandra , Tao JianQin , Feinstein Sheldon I. , Fisher Aron B. 
Published: May 2022

IntroductionALI is associated with bacterial infection (sepsis), either as an initiating factor or as a secondary complication. Activation of polymorphonuclear leucocytes (PMN) recruited into the lungs causes the release of free radicals and reactive oxygen species (ROS) that are responsible for many of the clinical manifestations of ALI. Since NADPH oxidase (NOX2) is the major source of ROS (i.e., superoxide anion) in PMN, NOX2 blockade has been proposed for the treatment of Acute Lung Injury (ALI). We reasoned that NOX2 blockers would be counter‐productive in ALI associated with bacterial infection by interfering with the bactericidal activity of PMN.HypothesisPhospholipase A2 inhibitory peptide (PIP‐2) that inhibits NOX2 activation (by preventing activation of Rac, a necessary NOX2 co‐factor) would inhibit the bactericidal activity of PMN and thereby limit clearance of bacteria from the lung; we posited that this effect of PIP‐2 could be overcome by the administration of appropriate antibiotics.MethodsUsing the cecal ligation and puncture (CLP) model of ALI associated with sepsis, we investigated the effect of inhibiting NOX2derived ROS production by administration of PIP‐2.ResultsThe number of colony‐forming bacteria in lungs and peritoneal fluid of mice with CLP approximately doubled at 18 h after intravenous treatment with 2 µg PIP‐2 /gram body weight (wt). Treatment with 10 µg PIP‐2/g body wt resulted in 100% mortality within 18 hr. Antibiotic treatment abolished both the increase in lung bacteria with low dose PIP‐2 and the increased mortality with high dose PIP‐2. Treatment with PIP‐2 in the presence of antibiotics resulted in significantly improved lung histology, decreased PMN infiltration, decreased lung fluid accumulation, and decreased oxidative lung injury compared to antibiotics alone.ConclusionAdministration of PIP‐2 provides partial protection against lung injury in a model of ALI due to bacterial infection while concurrent antibiotic treatment abolishes the deleterious effects of PIP‐2 on lung bacterial clearance. These results suggest that addition of PIP‐2 to the antibiotic regimen would be beneficial for treatment of ALI associated with bacterial infection.
Hepatic Lipidomic Analysis of Dietary Different Type of Fats and Fructose Interactions in the Pathogenesis of Nonalcoholic Fatty Liver Diseases
Authors: Shi Hongxue , McClain Craig , Song Ming 
Published: May 2022

ObjectiveDietary nutrient interactions are key determinants to the development of nonalcoholic fatty liver disease (NAFLD). Fat and fructose are two major components of Western diet. This study aimed to determine the combined effects of different type of dietary fats and fructose on the development of NAFLD in a murine model.MethodsMale adult C57BL/6J mice were assigned to six groups and fed with diets (AIN‐93G based) enriched with different types of fat, such as saturated fat (SF‐beef tallow), n‐6 polyunsaturated fat (n6USF‐corn oil), or n‐3 polyunsaturated fat (n3USF‐menhaden fish oil), respectively, with or without fructose supplementation through 10% fructose (w/v) in the drinking water ad lib for 20 weeks. 42% of calories was derived from the fat. Metabolic phenotypes were characterized by liver histology, plasma ALT and AST, glucose tolerance test, hepatic mRNA expression of genes involved in lipid metabolism, and lipidomic analysis of hepatic fatty acid composition.ResultsWhile both SF and n6USF fed mice developed obesity and hepatic steatosis with or without fructose, SFA fed mice were characterized with hepatomegaly, liver injury and fibrosis, which were further exacerbated by fructose; conversely, plasma triglyceride levels were lower compared to controls. Interestingly, n6USF fed mice exhibited steatosis without hepatomegaly, and this was associated with markedly elevated plasma triglyceride and free fatty acid levels, which were blunted by fructose. Moreover, fructose and n6USF led to markedly elevated glucose intolerance tests compared to controls. Hepatic lipidomic analysis revealed that total monounsaturated fatty acids, but not total SFA, exhibited a stepwise decrease from SF to n6USF and n3USF fed mice across most of the lipid pools, which parallels the hepatic triglyceride content, irrespective the presence of fructose. In line with this, hepatic mRNA levels of lipogenesis genes, including Srebf1, Scd1, Fasn, Acacaand Acacb, displayed a similar pattern of alterations (SF>n6USF>n3USF), suggesting that increased lipogenesis contributes to hepatic steatosis in SF and n6USF fed mice. Of note, fructose play a mild, but significant role in the alterations of specific individual fatty acids in certain fatty acid pools, despite the predominant role played by the dietary fats.ConclusionOur data suggest that excess energy from SF intake results in fat storage in the liver, likely due to impaired triglyceride secretion; whereas excess energy from n6USF intake is stored in the periphery. Both effects were exacerbated by fructose supplementation. n3USF is beneficial even in the presence of fructose.
Effects of Methanolic Extract of Moringa oleifera on an In Vitro Model of Diabetic Nephropathy
Authors: Tsotakos Nik , Mitra Debashree , Castaneira Ryan , Morris Daniel , Byler Josiah J. 
Published: May 2022

Diabetic nephropathy (DN) is a severe diabetic complication and a leading cause of chronic kidney disease and renal failure worldwide. The high prevalence of end stage renal disease (ESRD) due to diabetes, coupled with the lack of efficient treatments, prompts the need for newer therapeutic strategies that overcome the drawbacks of the existing ones. In recent years, Moringa oleiferahas been identified as a plant with multiple nutritional and medicinal advantages. Importantly, previous studies have reported antioxidant and anti‐hyperglycemic properties of Moringa leaf extracts in diabetic rats. The growing interest in the potential use of nutraceuticals over conventional drugs along with the recent evidence of the beneficial effects of M. oleifera extract led us to investigate the effects of the methanol extract of M. oleifera on an in vitro model of human glomerular epithelial cells (HGEC).HGEC were chronically cultured in the presence of physiological (5mM) or high glucose (25mM) levels and treated with different concentrations of methanolic extract of M. oleifera(MOE). We evaluated the effect of MOE on the production of advanced glycation end‐products (AGE), cell viability, and response to oxidative stress. Our results indicate that MOE can effectively inhibit AGE formation in vitro and provide a cytoprotective effect to cells following an oxidative challenge. However, in line with prior data indicating that HGEC chronically exposed to high glucose are primed for apoptosis, these cells showed reduced viability following exposure to MOE compared to cells exposed to normal glucose. These results suggest that acute, high doses of MOE might not be beneficial following chronic exposure of kidney cells to glucose, but the possibility of a protective effect of MOE when administered early is worth exploring.
Synthesis of MUC1 Peptide Backbone Bearing Sialyl‐Tn Antigen for Structural and Functional Studies With Endogenous Lectins
Authors: Ayyalasomayajula Ramya , Cudic Mare 
Published: May 2022

ObjectiveTo synthesize the tumor associated analogs of MUC1 for use in structural and functional studies between sialyl‐Tn (sTn) antigen with macrophage galactose‐specific lectin (MGL).AbstractMucin 1 (MUC1) is a heavily glycosylated transmembrane protein, altered in both expression and glycosylation pattern in human carcinomas of the epithelium. The presence of incomplete or truncated glycan structures, often capped by sialic acid, commonly known as tumor‐associated carbohydrate antigens (TACAs), on the cell surface is a well‐known cancer biomarker. Accumulating evidence suggests that TACAs expression is associated with tumor escape from immune defenses through interaction with endogenous carbohydrate binding proteins (lectins). These interactions often lead to creation of a protumor microenvironment, favoring tumor initiation, progression, metastasis, and immune evasion. Macrophage galactose binding lectin (MGL) is a C‐type lectin receptor found on antigen‐presenting cells (APCs) which facilitates the uptake of carbohydrate antigens for antigen presentation, modulating the immune response homeostasis, autoimmunity, and cancer. Considering the crucial role of tumor‐associated forms of MUC1 with MGL in tumor immunology, a thorough understanding of this interaction is essential for it to be exploited for cancer vaccine strategies. The specific aim of this research is to synthesize structurally well‐defined chemical probes, mono and multiple glycosylated MUC1 glycopeptide models carrying the Tn or sTn, that allow for the control of the complexity of the chemical space of the multivalent ligands. For this purpose, a concise scheme was developed for the large synthesis of the sTn antigen building block. Thiophenyl glycoside donors, in the presence TfOH/NIS or TMSOTf/NIS as promoter system, were used for the galactosylation and sialylation steps of the amino acid building block synthesis, respectively. We explored the effect of activator, temperature, solvent, and excess equivalent of sialic acid thioglycoside donor on sialylation reaction. The most favorable reaction conditions improved the desired α‐stereoselectivity, decreased the competing 2,3‐elimination reaction on the sialyl donor during activation, leading to overall increase in the reaction yield. This building block was used in the stepwise solid‐phase peptide synthesis of MUC1 glycopeptide models. The secondary structure of synthesized peptides was determined by circular dichroism (CD) spectroscopy.
Proteomics‐informed physiologically‐based pharmacokinetic (PBPK) modeling revealed differential effects of UGT2B17 variability on the pharmacokinetics of diclofenac following intravenous and oral administration
Authors: Ahire Deepak S. , Prasad Bhagwat 
Published: May 2022

Uridine 5′‐diphospho‐glucuronosyltransferase 2B17 (UGT2B17) demonstrates dramatic inter‐individual variability (>3000‐fold) in the activity and expression in humans, which is associated with age, sex, genotype, and ethnicity.1We recently found that diclofenac is a UGT2B17 substrate, however, the quantitative contribution of UGT2B17 in the metabolism and pharmacokinetics (PK) of diclofenac is not well studied. Moreover, we hypothesized that the higher intestinal abundance of UGT2B17 could further amplify the magnitude and variability in its in vivo PK. We first investigated diclofenac glucuronide (DG) formation kinetics in human liver microsomes (HLM) and human intestinal microsomes (HIM) that were isolated from human donors containing the deletion genotype and the highest‐expressor of UGT2B17 in our tissue bank. Diclofenac showed 4‐ and 16‐fold difference in maximum DG formation rate (Vmax) in UGT2B17 highest‐expressor versus deletion in HLM and HIM, respectively (Fig. 1). Proteomics‐informed PBPK modeling2 estimated the quantitative contribution (fm) of UGT2B17 in the intestinal and hepatic glucuronidation of diclofenac, which revealed fm of 0.77, 0.24, and 0.0 in the highest, average, and null UGT2B17 expressors, respectively. Interestingly, intestinal fm of UGT2B17 was 0.94, 0.90, and 0.0 in the highest, average, and null expressors, respectively. The organ‐specific changes in the fm were also substantiated by the predicted fraction gut escaped (fg), which was primarily associated with UGT2B17 abundance in the enterocytes. A PBPK model was first developed and validated in the UGT2B17 average expressor and then extended to other populations utilizing UGT2B17 abundance data. Predicted plasma levels (AUC) of diclofenac were 20 % and 100 % higher following an IV dose (50 mg) in UGT2B17 deletion versus the average and the highest expressor subjects, respectively. Similalry, after 50 mg oral dose, ~2‐fold and 8.5‐fold higher AUC was observed in UGT2B17 deletion as compared to that in the average and the highest UGT2B17 expressors, respectively. Sex differences in UGT2B17 abundance explained ~2‐fold higher diclofenac AUC in females after oral administration. In summary, the unique intestinal abundance and the highly prevalent gene deletion of UGT2B17 could likely contribute to the variable bioavailability of its substrate drugs. Similarly, poor metabolism of diclofenac in children and in UGT2B17 deletion subjects could predispose these patients to high GI discomfort.References

Bhatt et al., Drug Metab. Dispos. (2018).
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The activation status of integrin β1 determines the ACE‐2 expression and functions in renal epithelial and cancer cells
Authors: Mia Md Saimon , Shrestha Hum N. , Palamuttam Riya J. , Kumar Bony D. , Mathew Sijo 
Published: May 2022

BackgroundIntegrins are heterodimeric cell surface receptors that participate in intracellular signaling and regulate multiple cellular functions. These membrane receptors are also crucial for the organ development, structural maintenance, and disease pathologies of the kidney. Integrin β1 is the major integrin subunit present in the kidney. Alteration of integrin β1 expression is a characteristic of many kidney diseases such as kidney cancer and chronic kidney disease. The purpose of this study is to find the importance of integrin β1 in the expression and function of angiotensin‐converting enzyme 2 (ACE‐2) in the kidney. We hypothesized that the expression and function of ACE‐2 depend on the activation status of integrin β1 in the renal epithelial and cancer cells.MethodKidney epithelial‐specific integrin β1 KO mice were generated by crossing between ITGBfl/fland γGT‐Cre mice. PCR‐based analysis of genomic DNA was used to confirm the gene deletion. Expression of integrin β1 in the kidney cortex was monitored by qRT‐PCR. Immunoblotting and qRT‐PCR were performed to observe protein expression and mRNA level of ACE‐2 in mouse kidneys. The pharmacological compound, BTT 3033, was used to inhibit integrin β1 in HK‐2 and Caki‐1 cells. Integrin β1 activation status was monitored by immunocytochemistry using a conformation‐specific antibody. SARS‐CoV‐2 spike protein was used to find out the functional importance of the integrin β1‐ACE‐2 complex. Statistical significance of the observed data was carried out using a two‐tail t‐test.ResultsDeletion of integrin β1 from the renal epithelial cells reduced expression of ACE‐2 in the mice kidneys (n=3; p<0.05). Immunoblotting and qRT‐PCR showed, expression of integrin β1 (n=3; p<0.05) and ACE‐2 (n=3; p<0.05) were higher in renal cancer cells than renal epithelial cells indicating a positive correlation in their expression patterns. With the treatment of BTT 3033, there was a significant decrease in the activation status of integrin β1 in kidney epithelial (HK‐) and cancer (Caki‐1) cells (n=30; p<0.05). ACE‐2 mRNA levels decreased in HK‐2 and Caki‐1 cells when treated with an integrin β1 antagonist, BTT 3033 (n=3; p<0.05). Treatment of HK‐2 and Caki‐1 with BTT 3033 also decreased the internalization of the SARS‐CoV‐2 spike protein.ConclusionResults from this study demonstrate the importance of matrix proteins receptor integrin β1 in maintaining the ACE‐2 expression and function in the kidney. This study will have implications in various chronic diseases where significant changes in the composition of matrix proteins and integrin expressions are observed. Further studies are needed to understand the role of altered matrix composition in ACE‐2 mediated physiological functions.
Maternal Western‐style diet and obesity during pregnancy reduces mitochondrial Complex I‐linked oxidative capacity in tandem with decreased oxidative stress in skeletal muscle of adolescent Japanese macaque offspring
Authors: Greyslak Keenan T. , Hetrick Byron , Carey Kat , McCurdy Carrie E. 
Published: May 2022

BackgroundExposure to an obesogenic gestational environment during early development increases offspring risk of developing obesity and cardiometabolic diseases. Skeletal muscle of obese and insulin resistant individuals show signs of mitochondrial dysfunction including reduced oxidative capacity and increased oxidative stress. Subsequent oxidative damage is caused by reactive oxygen species (ROS) generated primarily by mitochondrial Complex I (CI). Previous work in our non‐human primate model shows decreased oxidative capacity in fetal skeletal muscle from obesogenic pregnancies, a phenotype we have confirmed is retained into adolescence. The purpose of this study was to evaluate how this reduced oxidative capacity correspond with ROS production in skeletal muscle of lean adolescent offspring.Experimental DesignLean (body fat <25%) adult female Japanese macaques were maintained on a control diet (CD) or Western‐style diet (WD) prior to and throughout pregnancy and lactation. Dams chronically consuming WD with body fat >30% were classified as obese and included in this study. Male and female offspring were weaned to CD and moved to independent housing at 7 months of age. Muscle samples were collected from offspring gastrocnemius (gastroc) and soleus at 40 months. Offspring data were analyzed by Student’s t‐test.ResultsExposure to maternal WD during pregnancy showed moderate but significant reductions in oxidative damage and a modest (but insignificant) decrease in ROS generation in gastroc of offspring from obese pregnancies. In the soleus, ROS production was significantly reduced in these offspring. Both nuclear‐ and mitochondrial‐encoded genes for CI were decreased with maternal obesity/WD pregnancy, indicating transcriptional regulation of CI in exposed offspring. Finally, CI protein abundance was also decreased in primary myotubes from these offspring in lieu of additional metabolic challenges, providing further evidence of lasting metabolic reprogramming in skeletal muscle.ConclusionsWe find that early life exposure to maternal WD‐induced obesity leaves a lasting impression on offspring skeletal muscle metabolism that persists into young adulthood. It is possible that transcriptional suppression of CI activity is a protective adaptation to mitigate oxidative damage caused by mitochondrial ROS. While long‐term consequences of altered oxidation patterns relating to ROS flux require further investigation, the loss of CI content and diminished oxidative capacity in these lean offspring likely drive their increased susceptibility for cardiometabolic diseases – particularly when faced with metabolic stress and environmental insults throughout the lifespan.
Alcohol Consumption Pattern and Nutritional Intake: Analysis of the 2017‐2018 National Health and Nutrition Examination Survey
Authors: Zirnheld Kara , Warner Jeffrey , Huirong Hu , Floyd Alison , Kong Maiying , McClain Craig , Kirpich Irina 
Published: May 2022

BackgroundPoor nutrition is often considered a component of alcohol use disorder as heavy drinkers substitute calories from macronutrients with ‘empty’ calories from ethanol. Indeed, the relationship between alcohol use and nutrition has long been of research interest due to the key role of diet in modifying individual susceptibility to alcohol‐induced multi‐organ pathology, such as alcohol‐associated liver disease. The goal of the current study was to evaluate nutritional intake among individuals with various drinking patterns through analysis of the 2017‐2018 National Health and Nutrition Examination Survey (NHANES). We hypothesized that individuals with the greatest alcohol intake would have the highest total caloric consumption, but that these individuals would have the lowest nutrient density in their overall diet.MethodsUsing the 2017‐2018 NHANES, we divided survey participants into the following four drinking categories based on guidelines from the National Institute on Alcohol Abuse and Alcoholism and the United States Department of Agriculture: never drinkers (n=500), non‐drinkers (past‐year abstainers, n=874), moderate drinkers (1 or 2 drinks per day for females or males, respectively, with no binge drinking, n=2658), and heavy drinkers (>1 or >2 drinks per day for females or males, respectively, or binge drinking, n=393). We analyzed 24‐hour dietary recall variables including macro‐ and micronutrient intake, as well as anthropometric measurements. One‐way ANOVA was used to determine significance by p values <0.05.ResultsCompared to never, non‐, and moderate drinkers, heavy drinkers consumed the highest total calories (by roughly 500 calories), coming from higher intake of carbohydrates, protein, fat, and alcohol. Further, heavy drinkers also consumed more of most micronutrients reported by the NHANES (apart from several antioxidant carotenoids such as alpha‐ and beta‐carotene and lutein/zeaxanthin). Interestingly, despite increased alcohol consumption and macro‐/micronutrient intake, heavy drinkers did not have an increased body mass index or waist circumference compared to other drinking groups.ConclusionThough poor nutrition has often been associated with excessive alcohol consumption, the results of this study showed that heavy drinkers consumed the most nutrient‐dense diet overall. Despite significant differences in total energy intake in heavy drinkers, anthropometric measurements were comparable across drinking groups. Future research should further investigate the relationship between alcohol use and nutrition in the NHANES and other sources.