bluuebunny/crossref_metadata_2025_split huggingface dataset:
Adipose‐tissue specific deletion of TGF‐β receptor 1 kinase protects mice from diet‐induced obesity and diabetes (577.4)
Authors: Wankhade Umesh , Wiese Mohmmad , Kulkarni Ashok , Rane Sushil 
Published: June 2021

The adipose tissue depot is one of main organs targeted during pathogenesis of diabetes and obesity. We previously showed that the TGF‐β/Smad3 signaling pathway regulates the appearance of brown adipocyte features in classical white adipose tissue (WAT). Global Smad3 knockout mice exhibit a phenotypic transition and functional transformation of WAT to BAT‐like features that provide protection to the mice from obesity and diabetes (Yadav et al 2011, Cell Metabolism). We also observed increased circulating levels of TGF‐β ligand in obese mice and human obese subjects suggesting an involvement of TGF‐ β receptors in the WAT‐BAT transition and regulation of glucose metabolism. To examine this possibility, we have generated an adipose tissue specific TGF‐β receptor 1 kinase knockout mouse model (TβRI‐KO). Epididymal WAT (EWAT) from TβRI‐KO mice showed increased BAT mRNA transcripts. Further, stromal vascular fractions (SVF) and pure adipocyte fractions from T[[Unsupported Character ‐ Symbol Font □]]RI‐KO mice showed increased levels of BAT progenitor markers. High fat diet (HFD) fed TβR1‐KO mice gain less weight, exhibit leaner body composition and improved glucose tolerance. EWAT and anterior subcutaneous WAT (AWAT) depots show smaller sized adipocytes with UCP‐1 stained multi‐locular cells interspersed in the WAT that resemble brown adipocytes. Consistent with this, EWAT from TβR1‐KO shows elevated BAT gene transcripts and significantly increased expression of BAT progenitor markers. Together, these results suggest that the TGF‐β receptor 1 kinase plays a vital role in adipose tissue biology by promoting its morphological and functional transformation to a metabolically beneficial brown fat like tissue with protective functions against obesity and diabetes.Grant Funding Source: Supported by NIDDK/NIH
In vitro analysis of novel inhibitors for Escherichia coli MurA (582.4)
Authors: Haile January , Frost Andrea , Boone Benjamin , Song Vanessa , Muzyka Jennifer 
Published: June 2021

The widespread of use of antibiotics to treat bacterial infection has selected for antibiotic resistant strains of bacteria. Fosfomycin is an example of a broad spectrum antibiotic that has become less effective because of increased bacterial resistance. Fosfomycin inhibits synthesis of peptidoglycan, a structural component of the bacterial cell wall. Fosfomycin covalently binds to the phosphoenolpyruvate binding site of the enzyme MurA, an essential enzyme in the peptidoglycan synthesis pathway. MurA catalyzes the addition of the enolpyruvate to UDP‐N‐acetylglucosamine. The additional product, an inorganic phosphate, is detected by the indicator, malachite green. Muzyka, et al. has identified putative inhibitors through DOCK and AMBER studies. IC50 values have been determined for BCB33b and VS5 of these molecules. Additional molecules with similar functional groups are being synthesized and evaluated as putative inhibitors. The site of inhibitor binding will be determined with in silico studies and site‐directed mutagenesis.Grant Funding Source: Centre College Faculty Development Fund and the Alcock Interdisciplinary Fund
Isolation and genomic annotation of two novel bacteriophage known to infect Mycobacterium smegmatis (589.5)
Authors: Paschalis Valerie , Patel Shivani , Aziz Froogh , Chen Alston , Monsen Kirsten , Adams Sandra , Vega Quinn 
Published: June 2021

Tuberculosis (TB), a disease caused by Mycobacterium tuberculosis, kills over 1 million people annually worldwide. With the development of multiple drug resistant strains of TB, interest has increased in the development of alternative methods for controlling this disease. As part of the HHMI‐SEA PHAGES project, bacteriophages capable of infecting Mycobacterium smegmatis, an organism in the same genus as M. tuberculosis, were isolated from soil samples in New Jersey. The phages were characterized by studying plaque morphology and by the use of Transmission Electron Microscopy. In addition, novel mycobacteriophages ShiVal and Alsfro were isolated and sequenced. The phage genomes were then annotated using DNA Master, BLAST, Glimmer, and GeneMark. Lastly, the information obtained was published in PhagesDB and GenBank. ShiVal is a B cluster, B1 subcluster phage with a genome size of 68K, 101 putative genes and 66.5% GC content. Alsfro is an A cluster, A1 subcluster phage with a genome size of 52K, 101 putative genes and a 63.3% GC content. Discovering and cataloging these novel phages is significant because a database of novel phages will be available to be tested for activity against M. tuberculosis.Grant Funding Source: HHMI SEA‐PHAGE
Epigenetic regulation of OPA1 sensitizes hepatocellular carcinoma to sorafenib‐induced apoptosis (59.1)
Authors: Puszyk William , Dong Hui‐Jia , Zhao Frank , Shisgal Preston , Cabrera Roniel , Neslon David , Liu Chen 
Published: June 2021

The aim of this study is to identify epigenetic markers of DNA methylation for the Optic Atrophy gene OPA1, which is an indication of Sorafenib sensitivity in HCC. Sorafenib has been shown to act as a multi‐kinase inhibitor in other cancers, and is used in the treatment of un‐resectable hepatocellular carcinoma (HCC). Patient response to this therapy is variable. We have previously shown that treatment with Sorafenib does not inhibit the AKT pathway in hepatocellular carcinoma cells and instead induces rapid mitochondrial fragmentation, which is associated with the deregulation of mitochondria‐fusion‐related protein OPA1, thus inducing apoptosis. Exposure of HCC cells or isolated mitochondria to Sorafenib induces cytochrome c release. Our data indicate that siRNA‐mediated OPA1 knockdown significantly sensitizes HCC cells to Sorafenib‐induced apoptosis. Furthermore, Sorafenib inhibits HCC xenograft tumor growth in vivo, and does not induce apoptosis in healthy primary hepatocytes. Murine xenograft tumor tissue samples revealed mitochondria fusion protein OPA1 expression levels are strongly associated with the sensitivity of HCC to Sorafenib treatment. We have shown that Sorafenib suppresses the tumorigenesis of HCC through the induction of mitochondrial injury via OPA1. We now show that epigenetic analysis of the OPA1 gene promoter indicates a role for DNA methylation in the suppression of OPA1 expression, and a potential biomarker which can be utilized for the identification of patients whom may benefit from treatment with Sorafenib. Our results provide new insights into the pathogenesis of HCC and suggest that OPA1 is a novel therapeutic target in patients with HCC.Grant Funding Source: NIH
The amyloid precursor protein of Alzheimer’s disease associates with axoplasmic organelles (596.10)
Authors: Stevenson James , Moran Sean , Cyr Jennifer , DeGiorgis Joseph 
Published: June 2021

The Amyloid Precursor Protein (APP) is a causative agent in Alzheimer’s disease (AD) as the Aβ fragment of this protein builds up in brain lesions of AD patients and mutations in APP lead to heritable forms of the disorder. APP contains a single transmembrane domain and a highly conserved region at the tip of the C‐terminus. Here, we show that APP associates with isolated axoplasmic organelles purified from the axoplasm of the squid giant axon. We have also identified the coding sequence of squid APP from a cDNA library of the North Atlantic Long‐finned Squid, Loligo pealei. We have cloned the full‐length coding sequence and are currently over‐expressing and purifying recombinant squid APP to establish a standard curve for quantitative Western blot to determine the concentration of APP in whole axoplasm and in isolated organelle fractions. Squid provide the only source of pure axoplasm as this organism contains a large axon up to 1 mm in diameter, allowing for the extrusion of cytoplasm away from the cell body, cell membrane, and other types of neuronal tissues. Quantifying the amount of APP in axoplasm and the number of APP molecules per organelle may provide insight into the wild‐type function of APP.Grant Funding Source: Supported by the Rhode Island INBRE Program NIMGS grant 5 P20 GM103430‐13
The highly conserved C‐terminus and transmembrane domain of the amyloid precursor protein of Alzheimer’s disease is found in highly divergent species (596.11)
Authors: O'Donnell Erin , Granato Juliana , DeGiorgis Joseph 
Published: June 2021

The Amyloid Precursor Protein is a causal agent in the pathogenesis of Alzheimer’s disease. A fragment of APP termed A‐beta builds up in the brain lesions of AD patients and mutations in the APP gene have been show to cause familial forms of the disorder. APP exhibits a single transmembrane domain that spans the lipid bilayer of membrane bound vesicles and a C‐terminal that contains a highly conserved sequence that extends into the neuronal cytoplasm. Here, we carried out a blast analysis of the highly conserved C‐terminal and then assayed hits for the presence of a transmembrane domain. Although, in some cases species have lost the homologous region of APP that causes AD (A‐bata) in humans, the C‐terminal and transmembrane domain are found in a wide array of organisms. Currently, we are using antibodies against the C‐terminal fragment to assay the presence and distribution of APP in distantly related species at the confocal and EM levels.Grant Funding Source: Supported by the Rhode Island INBRE Program NIMGS grant 5 P20 GM103430‐13
A new function for an old enzyme: Src protein tyrosine kinase directs excitotoxic neuronal death in stroke (596.3)
Authors: Cheng Heung‐Chin , Hossain Mohammed , Roulston Carli , Ng Dominic , Williamson Nicholas , Hoque Ashfaqul , Hill Andrew 
Published: June 2021

Excitotoxicity resulting from over‐stimulation of ionotropic glutamate receptors is a major cause of neuronal death in cerebral ischemic stroke. The over‐stimulated receptors exert their neurotoxic effects in part by over‐activation of calpains which induce neuronal death by catalyzing limited proteolysis of specific cellular proteins. Here, we report that in cultured cortical neurons and in vivo in a rat model of focal ischemic stroke, the tyrosine kinase Src is cleaved by calpains at a site in the N‐terminal unique domain. This generates a truncated Src fragment of ~52 kDa localised predominantly in the cytosol. A synthetic cell membrane‐permeable fusion peptide derived from the Src unique domain prevents calpains from cleaving Src in neurons and protects against excitotoxic neuronal death. To explore the role of the truncated Src fragment in neuronal death, we expressed a recombinant truncated Src fragment in cultured neurons, and demonstrated that expression of this fragment, which lacks the unique domain, was sufficient to induce neuronal death. Furthermore, inactivation of the pro‐survival kinase Akt is a key step in its neurotoxic signaling pathway. As Src maintains neuronal survival, our results implicate calpain cleavage as a key step of neuronal death in excitotoxicity, and suggest blockade of calpain cleavage of Src as a potential therapeutic strategy to minimize brain damage in ischemic stroke.Grant Funding Source: NHMRC Australia
Retrieval trafficking plays an important role in homeostasis of hERG expression at the plasma membrane (604.2)
Authors: Chen Jeffery , Guo Jun , Li Wentao , Yang Tonghua , Zhang Shetuan 
Published: June 2021

The human ether‐a‐go‐go‐related gene (hERG) encodes the α‐subunit of the rapidly activating delayed rectifier potassium channel (IKr). A reduction in hERG expression on the plasma membrane causes long QT syndrome, predisposing affected individuals to high risk of cardiac arrhythmias and sudden death. We previously reported that plasmalemmal hERG channels are vigorously internalized under certain conditions such as hypokalemia. Whether internalized channels can recycle back to the membrane remains unknown. In this study, using Western blot and patch clamp techniques, we observed that an enhanced recovery rate of IKr is associated with an increased endocytosis of hERG channels after exposure of cells to hypokalemic conditions. This observation is not due to increased synthesis since hERG mRNA expression was not increased by hypokalemic treatment and the enhanced recovery was not affected by the inhibition of protein synthesis using cycloheximide. Using fluorescent labeling, we directly monitored the retrieval trafficking of internalized hERG channels to the plasma membrane. Co‐immunoprecipitation experiments reveal that hERG channels are associated with the GTPase Rab11, but not Rab4. Interference with Rab11 function but not Rab4 led to decreased hERG expression and function. Our data provide novel insights into the recycling mechanisms that regulate the homeostasis of membrane‐bound hERG channels.Supported by the Canadian Institutes of Health Research ‐ CIHR
Intercellular adhesion molecule‐1 deficiency correlates with impaired inflammatory response in congenital disorder of glycosylation (607.2)
Authors: He Ping , Freeze Hudson 
Published: June 2021

Congenital Disorders of Glycosylation (CDGs) result from mutations in various N‐glycosylation genes. Many patients are prone to infections, but the mechanism is unknown. We found that CDG patient fibroblasts have less intercellular adhesion molecule‐1 (ICAM‐1), and because of its role in innate immune response, we hypothesized that its reduction may explain recurrent infections. In support of this, tunicamycin blocks TNF‐α induced ICAM‐1 production, suggesting that many CDG cells may show poor ICAM‐1 response. We, therefore, investigated a mouse model of MPI‐CDG, deficient in phosphomannose isomerase. We challenged MPI‐deficient mice with an intraperitoneal injection of pro‐inflammatory mediator (Zymosan) and found decreased neutrophil extravasation. Immunohistochemical staining of mesenteries showed attenuated neutrophil egress, presumably due to poor ICAM‐1 response to acute peritonitis. Since MPI‐CDG patients and their cells improve glycosylation when given mannose, we provided MPI‐deficient mice with mannose‐supplemented water for 7 days. This increased ICAM‐1 expression on mesenteric endothelial cells of Zymosan‐challenged mice and enhanced neutrophils exudation compared to untreated controls. Attenuated inflammatory response in glycosylation‐deficient mice may result from ICAM‐1 deficiency on the vascular endothelial surface and may help explain high mortality in patients.Grant Funding Source: supported by the Rocket Fund & NIH R01 DK55615
Regulator of G protein Signaling 6 suppresses Ras and DNA‐methyltransferase 1 expression and tumorigenesis in a mouse model of urinary bladder cancer (609.10)
Authors: Yang Jianqi , Platt Lance , Ibeawuchi Stella‐Rita , Stewart Adele , Maity Biswanath , Huang Jie , Fisher Rory 
Published: June 2021

Urinary bladder cancer, the fifth most common cancer in the U.S., frequently results from exposure to toxic chemicals including those present in cigarette smoke. A previous epidemiological study demonstrated that a polymorphism in RGS6 leading to increased translation/stability of RGS6 mRNA was associated with a pronounced reduction in the risk of bladder cancer, particularly in smokers. However, no studies have evaluated whether RGS6 functions as a tumor suppressor in bladder. Here, we employed RGS6‐/‐ mice to investigate the role of RGS6 in BBN (N‐butyl‐N‐(4‐hydroxybutyl) nitrosamine)‐induced bladder cancer, a model which matches well to its human counterpart. We found that RGS6 was highly enriched in mouse bladder, particularly in urothelium where transitional cell carcinoma originates. RGS6 loss was associated with increases in Ras and Dnmt1 expression in the bladder urothelium and marked decreases in Ras‐association domain family 1 isoform A (RASSF1A), a tumor suppressor and target of Dnmt1‐mediated silencing. A similar phenomenon was observed in BBN‐treated WT mice, an effect further exacerbated by BBN treatment of RGS6‐/‐ mice. In keeping with these observations, RGS6‐/‐ mice exhibited accelerated bladder tumor formation. Our findings suggest that RGS6 is a previously unrecognized, but critical repressor for bladder carcinogenesis.Grant Funding Source: Supported by NIH CA161882, AHA 11SDG7580008
A role for the tyrosine kinase Pyk2 in depolarization‐induced contraction of vascular smooth muscle (609.3)
Authors: Mills Ryan , Mita Mitsuo , Sutherland Cindy , Walsh Michael 
Published: June 2021

Background: Depolarization of vascular smooth muscle induces rapid (phasic) contraction followed by sustained (tonic) contraction. The latter is due to activation of the RhoA/Rho‐kinase pathway via genistein‐sensitive tyrosine phosphorylation.Objective: To determine if the Ca2+‐dependent tyrosine kinase Pyk2 is involved in depolarization‐induced sustained contraction.Methods: Depolarization of endothelium‐free rat caudal arterial smooth muscle was induced by high extracellular [K+] in the absence or presence of Pyk2 inhibitors. Isometric force was measured with a force transducer and Pyk2 activation (autophosphorylation of Y402) was quantified by western blotting with phosphospecific antibodies.Results: Depolarization induced Pyk2‐Y402 phosphorylation in a time‐dependent manner. Pyk2 inhibitors (sodium salicylate, PF431396 and others) prevented Y402 phosphorylation and inhibited K+‐induced contraction. Vanadate (tyrosine phosphatase inhibitor)‐, but not calyculin A (Ser/Thr phosphatase inhibitor)‐induced contraction was also inhibited by Pyk2 inhibition.Conclusion: Membrane depolarization induces phosphorylation and activation of Pyk2, which is required for the tonic phase of K+‐induced contraction.
Low density lipoprotein‐related protein 1 regulates lung inflammation (609.9)
Authors: Garcia‐Arcos Itsaso , Cummins Neville , Dabo Abdoulaye , Pillai Manju , Geraghty Patrick , Goldberg Ira , Foronjy Robert 
Published: June 2021

The low density lipoprotein related protein 1 (LRP1) is a widely expressed transmembrane receptor that binds a large number of ligands‐ApoE being one of them‐and is involved in inflammatory signaling cascades in multiple cell types. Recently, LRP1 was linked with lung function in a genomic study, but the implications of this association are unknown. We aimed at describing the role of LRP1 in lung inflammation. Mice were treated with the LRP1 ligand COG133, an ApoE mimetic peptide, for 1 month and they were compared with vehicle‐treated mice. The treatment with COG133 increased whole‐lung baseline protein phosphatase 2A (PP2A) activity, which is considered a defense mechanism often observed after cigarette smoke exposure, and decreased the mRNA expression of TNFα and IL8 in bronchoalveolar cells. In cultured lung epithelial cells, silencing LRP1 increased mRNA expression of MCP1, IL1β and TNFα; activated the transcription factors STAT1 and STAT3; and prevented the activation of PP2A in response to a challenge with cigarette smoke extract. In conclusion, LRP1 can modulate the expression of inflammatory cytokines and the activation of interferon‐mediated signaling cascades in lung.Grant Funding Source: Supported by NIH 5R01HL098528‐05
NODs‐dependent OmPGRP‐L1 expression modulates the excessive inflammatory response to bacterial infection in rainbow trout liver cells (611.1)
Authors: Jang Ju Hye , Kim Hyun , Kim Yu Jin , Cho Ju Hyun 
Published: June 2021

Peptidoglycan recognition proteins (PGRPs) recognize bacterial peptidoglycan and function in antibacterial innate immunity. OmPGRP‐L1, a long form of PGRP from the rainbow trout, is known to have an anti‐inflammatory role in liver cells. In this study, we examined the regulation of the expression of OmPGRP‐L1 in a rainbow trout hepatoma cell line RTH‐149 upon stimulation with microbial ligands. γ‐D‐glutamyl‐meso‐DAP (iE‐DAP) and muramyl dipeptide (MDP) markedly increased the expression of OmPGRP‐L1, as well as pro‐inflammatory cytokines. Silencing of nucleotide‐binding oligomerization domain‐containing protein (NOD)1 and NOD2 specifically inhibited the upregulation of OmPGRP‐L1 expression induced by their cognate ligands. Suppression of NF‐κB activation prevented the induction of OmPGRP‐L1 expression induced by iE‐DAP and MDP. In silico analysis and electrophoretic mobility shift assay revealed that the promoter of OmPGRP‐L1 had NF‐κB binding sites, suggesting that OmPGRP‐L1 is produced through NODs‐NF‐κB signaling pathway. Loss‐of‐function and gain‐of‐function experiments indicated that OmPGRP‐L1 down‐regulated the induction of pro‐inflammatory cytokine expression induced by iE‐DAP and MDP. Overall, these findings suggest that rainbow trout liver cells may modulate the excessive inflammatory response to bacterial infection by producing OmPGRP‐L1 through NODs.Grant Funding Source: Supported by the National Research Foundation of Korea (NRF‐2011‐0015180 and NRF‐2013M3A6A8073556)
Use of mid‐upper arm muscle circumference to identify undernutrition in hospitalized adults in Hanoi, Vietnam (620.1)
Authors: Manders Aaron , Pham Thi Thu Huong , Nguyen Thi Lam , Nghiem Nguyet Thu , Tran Chau Quyen , Dinh Thi Kim Lien , Nguyen Quoc Anh , Henry Elizabeth , Young Lorraine , Apovian Caroline , Ziegler Thomas , Lenders Carine 
Published: June 2021

Objective: To determine the utility of MUAC to screen underweight hospitalized patients in a major Hanoi teaching hospital.Methods: Data from a one‐day study of patients 蠅19 years old admitted at Bach Mai Hospital, Hanoi, Vietnam (2010) were utilized. Data included actual weight and height, MUAC, age, sex, presence of edema, reason for admission, ward, and prior subjective weight loss (major, >10%) over six months. BMI was calculated and underweight defined as BMI < 18.5 (kg/m²; WHO standards). Receiver operating characteristic curve (ROC) analyses was performed.Results: A total of 420 inpatients were studied (51.4 ± 0.8 years old; 177 (42%) female. Patients were admitted for acute (n=174, 41%), chronic (n=186, 44%), elective (n=31, 7%), or other (n=29, 7%) reasons. Mean ± SD for BMI was 20.0 ± 0.2 kg/m² and for MUAC 24.5 ± 0.2 cm. Rates of underweight status, and moderate and severe, or severe thinness were 34%, 17%, and 8%. A total of 22% subjects exhibited edema and 15% reported major weight loss prior to admission. For underweight status, sensitivity of MUAC <23 cm was 71%, specificity 90%, (+) predictive value 79%, and (‐) predictive value 86%. Area Under Curve (AUC) for MUAC<23 cm was 0.81.Conclusion: MUAC <23 cm appears to be a good method to identify undernutrition in hospitalized adults in Vietnam (AUC 0.80‐0.90 is good). Further evaluation of MUAC in larger samples and association with hospital morbidity and mortality in Vietnam are needed.Grant Funding Source: Supported by the Abbott Fund