bluuebunny/crossref_metadata_2025_split huggingface dataset:
O‐GlcNAcylase (OGA): A Sugar Cleaver
Authors: Gallen Corey , Rios‐Rosales Anapaula , Jacus Isabella G. , Cobb Allison , Nashman Zarrar , Grant Raheim , Shor Audrey C. 
Published: June 2021

A basic component of learning undergraduate biology concepts is grasping the interplay between carbohydrates and proteins. O‐GlcNAcylation is a post‐translational modification that covalently attaches O‐linked N‐acetylglocosamine (O‐GlcNAc) moieties to target proteins. Two enzymes regulate O‐GlcNAcylation; O‐GlcNAc transferase catalyzes the addition of O‐GlcNAc to specific serine and threonine residues, and O‐GlcNAcylase (OGA) hydrolyses the monosaccharide from the substrate. The focus of this project was to design and build a physical model that illustrates the key functional features of human OGA. Two isoforms of the enzyme exist in humans; 1 and 3 which are expressed in the cytoplasm and nucleus, respectively. The cytoplasmic isoform includes a full length protein, while the nuclear isoform is shorter that lacks a C‐terminal acetyltransferase‐like domain and demonstrates reduced enzymatic activity in vitro. Human OGAs possess a fugitive substrate binding groove that is highly conserved through metazoans. Two aspartic acid residues 174 and 175 comprise the catalytic activity of the enzyme and are conserved across all domains. Residues significant to binding O‐GlcNAc are close to the sugar/catalytic machinery or occupy positions shown to undergo a conformational change upon inhibitor binding. Mutations targeting the substrate binding groove identified residues that are not required for hydrolysis, but serve roles in recognition and binding of substrate. The differences between the residues required for the turnover of these substrates may reflect a change in molecular movement in OGA sites. Further elucidation of this mechanism could help advance our treatment and/or prevention of diseases like neurological disorders and diabetes.Support or Funding InformationFunded in part by NSF‐DUE 1725940 for the CREST Project.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Validation of Arid1a as a Mammary Tumor Driver in Mice
Authors: Winters Marquita , Kartha Nithya , Schimenti John 
Published: June 2021

Distinguishing commonly deleted genes as drivers or passengers of human cancers are key to delineating mechanisms of tumor initiation. The “Chaos3” mouse model of spontaneous breast cancer carries a missense mutation in MCM4 (minichromosome maintenance 4), causing destabilization of the MCM2‐7 replicative helicase, which in turn causes elevated genomic instability. Approximately 80% of female mice homozygous for this mutation in the C3H/FeJ background develop mammary tumors with an average latency of 12 months. Genomic analysis of these tumors revealed that ~70% had monoallelic deletion of tumor suppressor Arid1a. Arid1a is a member of the SWI/SNF chromatin remodeling complex and is responsible for conferring specificity of SWI/SNF complexes to their targets. Previous studies in the lab have shown that introduction of a second Arid1a allele in monoallelically deleted, cultured tumor cells rescued the tumorgenic phenotype following transplantation into host mammary glands. We propose that deletion of Arid1a early in postnatal development is one driver of Chaos3 mammary tumors. Here, we will perform intraductal mammary injections of adeno‐cre virus into Chaos3 females bearing a floxed Arid1a allele. Loss of Arid1a has been implicated as a driver of tumorigenesis in many cancers such as clear cell ovarian carcinoma and endometrioid carcinoma, and is deleted in ~5–11% of human breast cancers. Identifying Arid1a as a driver of mammary tumors in Chaos3 mice would validate its potential role in development of sporadic breast cancer in people.Support or Funding InformationBreast Cancer Coalition of RochesterThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
CD8+ T CELLS REGULATE LIVER INJURY IN OBESITY‐RELATED NONALCOHOLIC FATTY LIVER DISEASE
Authors: Kennedy Arion , Pacheco Cristina , Washington Mary Kay , Hasty Alyssa 
Published: June 2021

Nonalcoholic Fatty Liver Disease (NAFLD) is ranked as one of the top causes of liver disease in Western countries. We found an elevation in CD8 staining in livers from obese patients with NASH or cirrhosis that positively correlated with an elevation in smooth muscle actin (SMA). To examine the role of CD8+ T cells in the development of NAFLD, we used an obese and hyperlipidemic mild NASH model, the low density lipoprotein receptor deficient (LDLR−/−) mice fed a western diet (WD). CD8+ T‐cells were elevated by 3.5‐fold in the livers of mild NASH mice. Similarly, hepatic CD8 gene expression was significantly increased, along with cytokines Tnf and Il10 as well as hepatic stellate cell associated markers. Isolated hepatic CD8+ T‐cells from the mild NASH mice, had increased secretion of IL‐10, TNFa, IL‐6, IL‐4 and IL‐2 compared to chow fed mice. Chronic treatment with antibody against CD8 led to significant reductions in the gene expression of Il10, Tnf, Il2, Il4, Acta2 and Tgfb. Depletion of CD8+ T cells also led to a reduction in the recruitment of macrophages to the liver. In contrast, adoptive transfer of hepatic CD8+ T cells from the mild NASH mice into chow fed mice significantly increased hepatic Acta2 expression. In co‐culture studies, isolated CD8+ T cells from mild NASH mice induced hepatic stellate cell activation. In a non‐obese choline deficient mouse model of aggressive NASH, depletion and knockdown of CD8 T cells did not impact liver inflammation or hepatic stellate cell activation. In conclusion, our findings suggest that mild NASH – induced by obese and hyperlipidemia, is characterized by CD8+ T‐cell infiltrations of the liver, subsequently regulating the recruitment of macrophages and activation of hepatic stellate cells.Support or Funding InformationThis work was supported by an American Diabetes Association Mentor‐Based Postdoctoral Fellowship (7‐10‐MI‐05), UNCF‐Merck Postdoctoral Science Research Fellowship, the Vanderbilt CTSA grant UL1 TR000445 from NCATS/NIH and K01HL121010 to AK. AHH is supported by a Merit Award from the Veterans Affairs (5I01BX002195), an Established Investigator Award from the American Heart Association (12EIA827), and an Innovation Award from the American Diabetes Association (1‐17‐IBS‐140). Confocal Microscopy was performed through the use of the VUMC Cell Imaging Shared Resource (supported by NIH grants CA68485, DK20593, DK58404, HD15052, DK59637 and EY08126). Flow Cytometry experiments were performed in the VMC Flow Cytometry Shared Resource, which is supported by the Vanderbilt Digestive Disease Research Center (DK058404) from whom AHH received scholarship funds.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
SIRT3 Activation Inhibits Development of Diabetic Kidney Disease
Authors: Myakala Komuraiah , Wang Xiaoxin , Lewien Pierce , Wang Dong , Luo Yuhuan , Herman‐Edelstein Michal , Rosenberg Avi Z. , Levi Moshe 
Published: June 2021

Diabetes continues to be the leading causes of kidney disease in the U.S. and around the world. Despite beneficial interventions implemented in patients with diabetes that mitigate some of its negative effects, kidney disease still progresses in most of these patients because the pathogenesis of diabetic kidney disease remains poorly understood. Recent evidence indicates that impairments in mitochondrial dynamics and function plays an important role in pathogenesis of diabetic kidney disease and brings promise to treatment strategies for improving mitochondrial function and the related pathways that may prevent or slow the progression of kidney disease.We aimed to test the influence of one key mitochondrial protein deacetylase, Sirtuin 3 (SIRT3) as a potential target for therapy in diabetic kidney disease. In kidney biopsies from human patients with diabetic nephropathy, there are decreases in SIRT3 mRNA and protein abundance in glomeruli and tubules. To test the effect of SIRT3 on the prevention of diabetic kidney disease, we conducted a study comparing three groups of mice: A control group, a diabetic group (db/db), and a diabetic group treated with nicotinamide riboside (NR), a precursor for NAD+, which is a co‐substrate required for SIRT3 activity. Treatment of diabetic mice with NR resulted in a) increase in renal SIRT3 activity as determined by significant decreases in total acetylated proteins and acetylated SOD2, significant decreases in b) albuminuria, c) mesangial expansion and glomerular area, d) accumulation of collagen IV protein, e) loss of synaptopodin and f) accumulation of 4‐HNE. In conclusion, SIRT3 plays an important role in diabetic kidney disease and increasing the SIRT3 activity may prevent the progression of diabetic kidney disease.Support or Funding InformationNIHThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Activator Protein 1 and Caspase 8 Mediate Palmitate‐Induced Cardiomyocyte Apoptosis
Authors: Oh Charles , Lee John , D'Souza Karen , Migrino Raymond , Thornburg Kent , Reaven Peter 
Published: June 2021

IntroductionLipoapoptosisof cardiomyocytes may be the basis for diabetic cardiomyopathy. Wepreviously demonstrated that p38α mitogen activated protein kinase (MAPK) mediatespalmitate (PA)‐induced cardiomyocyte apoptosis, and that specific p38α siRNAmarkedly and dose‐dependently attenuates apoptosis in cardiomyocytes treated with PA. We tested the hypothesis that p38α‐dependent pathway consists, in part, of c‐fos subunit of Activator Protein 1 (AP‐1) and caspase8, and that this p38α/AP‐1/caspase 8 cascade mediate PA‐induced cardiomyocyte apoptosis.MethodsHuman adult ventricular cardiomyocytes line (AC16 cells) were exposed to high physiological levels of PA. p38α‐dependent pathway was evaluated using p38α and c‐fos siRNA knockdown, and caspase 8 inhibitor.ResultsWith PA treatment, dose‐dependent increases in c‐fos mRNA (1.83 fold and 3.13 fold control, at 150 μM and 300 μM respectively, n=6, p<0.01), in c‐fos protein (1.00 fold and 2.67 fold, at 150 μM and 300 μM respectively, n=3, p<0.05), and a much greater increase in phospho‐c‐fos (2.64 fold and 8.94 fold, at 150 μM and 300 μM respectively, n=, p<0.01) were measured. Transcription of caspase 8 also increased (1.40 fold and 1.80 fold, at 150 μM and 300 μM respectively, n=5, p<0.05) but procaspase 8 protein level did not.p38α knockdown dose‐dependently blocked the PA‐induced increase in phospho‐c‐fos protein level(to 62%, 45%, and 30% of control (si‐control + PA 300 μM group) for 30 pmol, 60 pmol, and 120 pmol si‐p38α knockdown groups, n=5, p<0.01), but total c‐fos level did not change significantly. Interestingly, procaspase 8 level was also dose‐dependently reduced by p38α knockdown (75%, 65%, and 59% of control (si‐control + PA 300 μM group) for 30 pmol, 60 pmol, and 120 pmolsi‐p38α knockdown groups, n=5, p<0.05).With c‐fos knockdown, PA‐induced apoptosis was reduced 39% (n=3, p<0.05) 24 h post‐transfection, and by 53% after 48 h post‐transfection (n=3, p<0.01). Caspase 8 inhibition for 24 h reduced apoptosis by 39% (n=3, p<0.01).ConclusionsThis study in cardiomyocytes demonstrated that PA increases expression and phosphorylation of c‐fos subunit of AP‐1, and caspase 8 transcription, and further that p38α knockdown attenuates PA‐induced increases in c‐fos and phospho‐c‐fos, and decreases caspase 8. Translational knockdown of c‐fos attenuated PA‐induced cardiomyocyte apoptosis, as did inhibition of caspase 8. Our results support the hypothesis that PA‐induced apoptosis is mediated by p38α, AP‐1/c‐fos, and caspase8.Support or Funding InformationThis work has been funded by VA VISN 18 New Investigator Grant (to C.O.), CSR&D5I01CX000598grant (to P.R.), and VA Merit grant BLRDI01BX007080(to R.M.). The study was supported by VA Employment and Carl T. Hayden Medical Research Foundation. The contents and the views do not represent the views of the Department of Veterans Affairs or the US government.
Targeting and tracing cardiac myocytes with Fgf1 expression in F1A‐CreERT2 transgenic mice
Authors: Chiu Ing‐Ming , Chung Yu‐Fen , Chen Mei‐Shu , Jiang Si‐Tse , Hsu Yi‐Chao 
Published: June 2021

Fibroblast growth factor 1 (FGF1) regulates many biological and physiological processes. In mice, Fgf1 gene contains at least three upstream promoters and spliced to the first protein coding exon alternatively, giving rise to different Fgf1 mRNA variants (1A, 1B and 1G). Among them, the Fgf1A transcript is predominantly expressed in heart. FGF1 can induce cardiomyocyte regeneration and cardiogenesis in vitro and in vivo. Here, we generated a novel mouse line Fgf1A promoter (F1A)‐driven inducible Cre recombinase (CreERT2). We firstly demonstrated that the highest mRNA expression of CreERT2 were detected in the heart specifically of F1A‐CreERT2 founder mice, similar to that of Fgf1A mRNA. The F1A‐CreERT2 mice were crossed with ROSA26R mice, and the F1 mice were analyzed. The punctate blue LacZ‐positive signals were detected exclusively in the heart after tamoxifen administration. The CreERT2‐mediated recombination in the tissues is monitored through punctate blue LacZ‐positive signals, indicating the in situ localization of F1A‐positive cells. Interestingly, these F1A‐positive cells with punctate blue signals were colocalized with cardiomyocytes expressing a‐actinin, a‐myosin heavy chain and cardiac troponin T, suggesting cardiomyocyte‐specific expression of Fgf1A. Our data suggested that the F1A‐CreERT2 mouse line could be used for time‐dependent and lineage tracing of Fgf1‐expressing cells in vivo.Support or Funding InformationMinistry of Science and Technology, Taiwan;Ministry of Health and Welfare, Taiwan;National Health Research Institutes, TaiwanThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Understanding the role of pancreatic β‐cell CD36 in the development of Type 2 Diabetes
Authors: Kolar Elizabeth Anne , Gajrawala Eshaa , Deeney Jude , Hamilton James 
Published: June 2021

CD36 is a transmembrane scavenger receptor for oxidized LDL (oxLDL). Expression of CD36 and subsequent uptake of oxLDL into macrophages is central to foam cell formation in atherosclerotic plaques. CD36 is also reported to play a pivotal role in glucotoxicity‐induced β‐cell dysfunction in Type 2 Diabetes (T2D). The aim of this study is to investigate mechanistic functions of CD36 that contribute to β‐cell dysfunction and the onset of T2D. For this study, rat INS‐1 pancreatic β‐cells were cultured in either low (5 mM) or high (11 mM) glucose conditions. Cells grown in high glucose exhibited a left shift in glucose stimulated insulin secretion (GSIS), showing that the cells are more sensitive to lower glucose levels. These cells also contained more lipid droplets, as shown by Nile Red staining. Using immunofluorescence and Western blotting, we demonstrated that CD36 expression increased with increasing glucose concentrations. When cells were incubated with DiI‐labeled oxLDL to investigate uptake, cells that were grown in high glucose exhibited increased labeling. Interestingly, oxLDL uptake by INS‐1 cells grown in 11 mM glucose can be suppressed by the addition of the ω‐3 fatty acid, docosahexaenoic acid (DHA). Studies investigating the effect of DHA and additional fatty acids on uptake of oxLDL by INS‐1 cells and effects on GSIS are ongoing. We propose that CD36 plays a role in how β‐cells respond to high glucose levels, similarly found in T2D. The increase in oxLDL uptake via CD36 in these cells may contribute to glucolipotoxic effects in β‐cells that results in aberrant insulin secretion in T2D.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Designing a Model Skin Cell Membrane to Investigate the Extent of Nanoparticle Absorption
Authors: Yoder Brandi , Sostarecz Audra Goach 
Published: June 2021

The Langmuir Monolayer Technique is a useful method for modeling the epithelial cell membrane and for investigating the organization of membrane molecules. Liposomes with fluorescence analysis are useful in interpreting molecule interactions with the hydrophilic and hydrophobic regions of the membrane. Zinc oxide particles were introduced to monolayers of lipids found in the stratum corneum and also to liposomes of the same composition to investigate how these particles interact within the cell membrane. This process will allow us to ultimately determine if the nanoparticles in over the counter beauty products embed our skin. In our lab, the compression isotherms of four lipids, cholesterol, cholesterol sulfate, lignoceric acid, and N‐oleoyl‐d‐sphingosine, which are major components of the epithelial cell membrane have been established at body temperature conditions. Ceramide molecules have also been analyzed and incorporated to create a more realistic model system. Liposomes were used as a technique to study the bilayer due to the ability to analyze them via fluorescence spectroscopy by incorporation of a fluorescent ceramide. The ultimate goal is to determine whether or not the addition of the zinc oxide will increase the fluidity of our membrane system utilized for modeling epithelial cells.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Harnessing glycocalyx interactions to modulate differentiation and development
Authors: Huang Mia L. , Michalak Austen L. , Tota Ember M. , Smith Raymond Alexander , Trieger Greg W. , Godula Kamil 
Published: June 2021

The cellular fate of embryonic stem cells and their subsequent development into differentiated cells are governed by precise growth factor signaling events at the stem cell surface. Such events are often orchestrated by heparan sulfate proteoglycans (HSPGs), which template growth factor binding to their corresponding receptors. Given the structural complexity of HSPGs, however, methods to manipulate them to study their function has been limited. Here, we report a suite of chemical strategies that exploit the function of HSPGs to control pluripotency or trigger the differentiation of mouse embryonic stem cells. To this end, we employ small molecules targeting the glycosaminoglycan (GAG) side chains of HSPGs to maintain pluripotency, and design GAG‐mimetic molecules that can recapitulate the functions of native GAGs.Support or Funding InformationNIH Pathway to Independence Award (1K99HD09029201, to MLH)NIH Director's New Innovator Award (1DP2HD08795401, to KG)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mapping Specific Cellular Sialoglycans Using Glycosyltransferases
Authors: Wu Zhengliang L. , Person Anthony D. , Burroughs Barbara , Singh Ravinder , Tatge Timothy , Manning Timothy , Wu Guoping , Sackstein Robert , Kalabokis Vassili 
Published: June 2021

The spatial and temporal decoration of glycans with neuraminic acid (sialylation) on cell surface involves in many biological processes, ranging from providing a physical barrier, shielding galactose and mannose residues from respective receptors, to generating specific epitopes that guide cell adhesion events. For examples, sialylated Lewis X structure serves as a ligand for selectins and is essential for leukocyte homing process; sialylated T/Tn antigens are involved in oncogenic processes and cancer invasiveness; while α‐(2,3)‐linked sialic acids serve as receptors for avian influenza vial infection, α‐(2,6)‐linked sialic acids serve as receptors for human influenza viral infection. Mapping the distribution of these glycans on cell surface is essential for us to understand their biological roles. Here, using HELA, HUVEC and CHO cells as models, we describe techniques for imaging some specific sialo‐glycans using glycosyltransferases including ST6Gal1, ST3Gal1, ST6GalNAc1, FUT2, and FUT9, with respective clickable nucleotide sugar donors. Our data suggest that while small percentages of terminal lactosamine (precursors for Lewis X structure) and Tn antigen may not be sialylated, almost all N‐glycans and Core‐1 O‐glycans on HELA cells are sialylated. Our data also indicate that while N‐glycan and lactosamine are more enriched on cell edges, where they may participate in cell‐cell interactions, sialylated Core‐1 O‐glycan and Tn antigen are more evenly distributed on cell surface. Despite the notion that CHO cells show no evidence for ST6Gal1 gene expression, we found that CHO cells are covered with α‐(2,6)‐linked sialic acid, which is also confirmed by sialic acid probing on CHO expressed proteins.Support or Funding InformationThis work was supported by R&D Systems/Bio‐techneThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Multi‐omics Studies Identify the Role of PPAR Pathway in Rheumatic Heart Disease Patients with Chronic Atrial Fibrillation
Authors: Li Ming‐Yang , Chen Huan‐Xin , Hou Hai‐Tao , Wang Jun , Yuan Chao , Yang Qin , He Guo‐Wei 
Published: June 2021

BackgroundAtrial fibrillation (AF) is one of the most common arrhythmia encountered in the clinical practice. The estimated prevalence of AF in the general population is as high as 1–2%. In this study, we aimed to identify the molecular changes and mechanisms responsible for chronic atrial fibrillation in rheumatic heart disease (RHD).MethodsPatients with rheumatic heart diseases (N=162) were enrolled in this study and classified into chronic AF group and sinus rhythm (SR) group based on their clinical manifestation. The atrial tissue (N=62) and plasma (N=100) samples were collected from the patients undergoing valve replacement surgery. Multi‐omics Studies were fulfilled with RNA Sequencing (RNA‐seq), isobaric tags for relative and absolute quantification (iTRAQ) and gas chromatography‐mass spectrometer (GC‐MS). ELISA was used to validate the proteins.ResultsThe differential genes/proteins were identified in the tissue or in the plasma samples between RHD‐AF patients and RHD‐SR patients. Combined analysis of transcriptomics, proteomics, and metabolomics revealed that the PPAR signaling pathway plays an important role in AF. The expression of multiple proteins were associated with PPARα,β,γ in tissue samples. On the other hand, only PPARα downstream alternations were identified in the study of plasma samples (APO‐A1 and APO‐A2). These proteins may be potential biomarkers for the diagnosis and treatment of AF in the future.ConclusionsWe for the first time identified the differentially expressed genes/proteins and related metabolites from plasma and tissue in chronic AF patients. Our study suggests that AF may impair the energy metabolism in the heart by altering the PPAR pathway, thus presenting metabolic remodeling in the heart.Support or Funding InformationSupported by grants from the National Natural Science Foundation of China (81641017), Zhejiang Provincial Natural Science Foundation (LY15H020008), & Tianjin Binhai Key Platform for Creative Research Program (2012‐BK110004), Binhai New Area Health Bureau (2014BHKY010, 2012BWKZ008, 2016BWKY007, 2016BWKZ003).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.