bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identifying Mechanisms of Interleukin‐19‐Mediated HuR‐Dependent Reduction in Vascular Cell mRNA Stability
Authors: England Ross Nevin , Kelemen Sheri , Ellison Stephen , Autieri Michael 
Published: June 2021

IL‐19 reduces TNF‐α‐driven mRNA abundance and protein abundance of intercellular adhesion molecule‐1 (ICAM‐1) and vascular cell adhesion molecule‐1 (VCAM‐1) (p<0.01) in cultured coronary artery endothelial cells (ECs) as measured by RT‐PCR and immunoblot. IL‐19 treatment produced no change in activation of the transcription factor NF‐κB, as measured by immunoblot analysis of p65 phosphorylation, translocation, and IkB degradation. IL‐19 decreases stability of ICAM‐1 and VCAM‐1 mRNA in actinomycin‐D treated cells. IL‐19 treatment inhibits cytoplasmic translocation of the mRNA stability factor HuR as assayed by immunoblot and immunofluorescence. siRNA knock‐down of HuR also decreases ICAM‐1 and VCAM‐1 mRNA stability. IL‐19 decreases HuR cytoplasmic translocation and phosphorylation as measured by phospho‐serine immunoprecipitation of HuR. In cultured coronary artery smooth muscle cells (SMCs) and ECs, IL‐19 increases expression of putative HuR regulator microRNA‐133, measured using the miScript PCR system (Qiagen), and treatment with miR‐133 mimic reduced expression of HuR mRNA.These data support our hypothesis that IL‐19 exerts its anti‐inflammatory effects by HuR‐mediated decrease in mRNA stability. We propose inhibition of HuR phosphorylation and translocation and upregulation of miR‐133 as possible mechanisms.
Modulation of the receptor ALX/FPR2 in vascular inflammation
Authors: Perretti Mauro , Bena Stefania , Brancaleone Vincenzo , Flower Roderick J 
Published: June 2021

The receptor ALX/FPR2 conveys the tissue protective properties of lipoxin A4 and Annexin A1 (AnxA1). Here, we tested if ALX was modulated during ischemia/reperfusion (IR) to attenuate secondary organ injury. A mouse colony nullified for ALX and with a GFP reporter was used. Acute myocardial infarct (25/120 min IR) was characterised by high mortality (~40%) in Alx null, but not wild type, mice. Using 90 min reperfusion revealed a higher degree of heart necrosis in the null mouse. In the lung, Alx promoter activity was evident in resting conditions, with signal in bronchial epithelia and scattered macrophages, and was markedly upregulated after IR (qPCR and immunohistochemistry). Higher KC, TNFα and IL6 levels were quantified in lung extracts of Alx null mice and were associated with augmented MPO activity values. These data indicate profound modulatory functions of ALX/FPR2 in IR with rapid up‐regulation and non‐reduntant functions in distant organs such as the lung.Supported by The Wellcome Trust
Differential expression of pro‐inflammatory and pro‐resolving lipid signaling genes in ARDS
Authors: Abdulnour Raja‐Elie Edward , Howrylak Judie A , Dolinay Tamas , Choi Augustine , Levy Bruce D 
Published: June 2021

RATIONALELipid mediators play important roles in the pathophysiology of the acute respiratory distress syndrome (ARDS), so we investigated the differential expression of genes associated with lipid signaling in patients with ARDS.METHODSFor patients that met clinical criteria for sepsis with ARDS (n= 19) and sepsis alone (n=29) whole blood was obtained on admission to the medical intensive care unit (day 0) and on day 7. RNA was extracted and microarray analysis performed to compare the expression profile of genes related to eicosanoid biosynthesis, function, and regulation between the two groups. Validation of select candidate genes was performed in experimental ARDS in BALB/c mice (50μl 0.1N HCl, pH 1.0 or saline) by semi‐quantitative PCR on lung RNA.RESULTSSeveral pro‐inflammatory lipid genes were upregulated at ARDS day 0 and downregulated at day 7 including PLA2G2A, ALOX5, ALOX5AP, PTGS2, PTGER3, and LTB4R. In contrast, genes linked to resolution displayed distinct patterns of expression; ALOX15, ALOX15B and PTGER4 were downregulated whereas AHR, GPR32 and ALX/FPR2 were upregulated. Genes involved in phosphatidic acid, sphingomyelin, and polyisoprenyl diphosphate metabolism were differentially regulated. For example, PLD1 and SGMS2 were upregulated at day 0 and downregulated at day 7, while PPAP2C and PPAPDC2 were upregulated at both time points. Analysis of gene expression in a murine model of acute lung injury confirmed selected target genes.CONCLUSIONThese findings emphasize that pro‐inflammatory and pro‐resolving lipid signaling genes are subject to temporally distinct regulation in the early phases of ARDS, and have uncovered several new potential points of regulation. The work is supported in part by the NIH (T32‐HL07633, R01‐HL068669, P01‐GM095467)
Identification of differentially expressed microRNAs in rat brain neuronal culture induced by Angiotensin II
Authors: Singh Neha , Guo Lirong , Zhong Tiecheng , O'Rourke Stephen T. , Sun Chengwen 
Published: June 2021

The brain renin–angiotensin system (RAS), particularly Angiotensin II (Ang II) plays an important role in the central control of blood pressure. However, the role of microRNAs (miRs) in this process is unclear. Our aim was to identify the miRs that are altered by Ang II treatment in rat brainstem and hypothalamus neurons. A genome wide miRNA array Quantitative PCR was performed and analyzed in neuronal cultures treated with or without Ang II (100 nM, 24h). Analysis of miRNA arrays revealed several miR levels were altered by Ang II (100 nM, 24h) as compared to control neurons. Five miRs that were most up‐regulated by Ang II were miR‐219–1‐3p (18.4 fold), miR‐466c (15.7 fold), miR‐136 (10.2 fold), miR‐27a* (10.1 fold), miR‐25* (10 fold); and those that were most down‐regulated were miR‐9* (139 fold), miR‐20b‐5p (89.5 fold), miR‐20a (83.6 fold), miR‐26a (74.3fold) and miR‐124 (74.1 fold). Several critical genes have been identified by bioinformatic analysis for the validated target genes of these miRs, such as cannabinoid receptors (CB1R), Neuregulin 1 and STK39, suggesting that Ang II might be acting through these differentially expressed miRs to regulate other gene expression. In this study, we identified several miRs altered by Ang II treatment, which may shift neuronal gene profile, contributing to the long‐term blood pressure regulation.Research Support: NDSU‐COBRE, ND‐EPSCoR
Interplay between complement factor B and Toll‐like receptors and its role in septic cardiomyopathy
Authors: Zou Lin , Li Yan , Feng Yan , Zhang Ming , Chen Chan , Chao Wei 
Published: June 2021

ObjectiveThe goal of the current study was to determine whether complement factor B (cfB) would interplay with Toll‐like receptors (TLRs) during sepsis and whether or not such interplay would contribute to septic cardiomyopathy.MethodsCardiomyocytes (CMs) were isolated from wild‐type (WT) mice or mice deficient of TLR2 or TLR4. Polymicrobial sepsis was created by cecum ligation and puncture (CLP). cfB mRNA and protein were determined by qRT‐PCR and Western blot, respectively. Cytokines were determined using a fluorescent multiplex assay. Left ventricular function was assessed in a Langendorff perfusion system.ResultsBoth Pam3cys and lipopolysaccharide (LPS) markedly increased cfB mRNA expression in CMs via TLR2‐ and TLR4‐dependent manner (fold, 217±54 and 578±54, respectively), but not complement 3, 4 or 5. TLR2, TLR3, TLR4 ligands also induced marked cfB protein expression in and release from CMs. In vivo, CLP led to a marked and time‐dependent increase in cfB mRNA and protein expression in septic heart via MyD88‐dependent manner. Importantly, compared with septic WT mice, septic cfB‐KO mice had improved LV function, better survival rate, and decreased cytokine production.ConclusioncfB is up‐regulated in response to TLR activation and polymicrobial sepsis via TLR‐dependent manner. cfB deficiency confers a beneficial effect in cardiac function during sepsis.This research has been supported by NIH R01‐GM080906 and R01‐GM097259.
Regulation of hypoxic responses by Sprouty2 in hepatocellular carcinoma
Authors: Hicks Kristin C , Patel Tarun B 
Published: June 2021

Sprouty2 (Spry2) is an inhibitor of receptor tyrosine kinases (RTKs) and decreases in its levels have been documented in breast, lung, liver, and prostate cancer. Decreases in Spry2 levels have been correlated with poor patient prognosis in hepatocellular carcinoma (HCC). During tumor growth, cells experience hypoxic periods and adapt to these by altering a variety of cellular processes mainly through the Hypoxia Inducible transcription Factors (HIFs). HIFs are composed of an α subunit (HIF1α or HIF2α) and a β subunit (HIF1β a.k.a. aryl hydrocarbon receptor nuclear translocator (ARNT)). In hypoxia, the HIFα subunits are stabilized, translocate to the nucleus, and dimerize with ARNT. The HIFs have been shown to be upregulated in cancers correlating to poor patient prognosis. We have shown that Spry2 protein levels are elevated in hypoxia. The objective of our study was to determine the role of Spry2 in regulating hypoxic responses in HCC cells as it could have an important impact in tumor growth and progression. We found that endogenous Spry2 by modulating ARNT protein levels alters the transcriptional activity of HIF. Thus, our data suggest that Spry2 regulates HIF activity in HCC cells.Supported by NIH grant GM 073181
Cannabinoid 1 receptor as therapeutic target in preventing chronic epilepsy
Authors: Di Maio Roberto 
Published: June 2021

ObjectiveTemporal lobe epilepsy (TLE) is the most common form of refractory epilepsy. Cannabinoid type 1 (CB1) receptor regulates neuronal excitability and has been shown to mediate the anticonvulsant effects of cannabinoids in acute animal models of seizure. However, the potential of cannabinoids for preventing chronic epileptic damage as consequence of an inciting event (acute seizure) remains under explored.MethodsWe investigated the neuroprotective role of the CB1 receptor against the hippocampal damage occurring in a chronic pilocarpine (PILO) model of TLE in rats. We simulated a therapeutic approach by administering the CB1 agonist WIN55.212–2 to rats for 15 days, beginning 24 hours after the induction of acute epileptic syndrome with PILO.ResultsAfter 6 months, the animals treated with WIN55.212–2 for 15 days following PILO administration, showed (relative to vehicle control): decreased epileptic behavior, reduced abnormal fiber sprouting in the dentate gyrus, preservation of GABA‐ergic neurons, less oxidative injury through NADPH oxidase activation, and normalization of CB1 receptor distribution and expression. Taken together, these data suggest that novel compounds targeting CB1 receptor or downstream signaling pathways may be efficacious in the prevention of chronic TLE.InterpretationGiven the prevalence and severity of TLE, there is great potential of such compounds.
The malin‐laforin compex modulates glycogen catabolism through its effect on glycogen phosphorylase
Authors: Dukhande Vikas V , Paasch Bradley , Gentry Matthew S 
Published: June 2021

Autosomal recessive mutations in genes encoding malin or laforin cause Lafora disease (LD). LD is a progressive myoclonic epilepsy characterized by pathological aberrant glycogen accumulations in brain and various other organs. Laforin is a novel glycogen phosphatase essential for maintaining glycogen in a soluble form. Malin, a RING‐type E3 ubiquitin ligase, is known to ubiquitinate proteins involved in glycogen metabolism. However, malin's role in LD pathogenesis is not clear. In this study, we aimed to discover malin substrate and define their function in LD pathogenesis. We used malin knockout and laforin knockout mice, immunoprecipitation, western analysis, immunocytochemistry, glucan‐binding assay, PAS staining and enzyme activity measurements. We discovered glycogen phosphorylase (GP) as a novel substrate of malin. GP is vital in catabolism of cellular glycogen. The expression of GP was decreased in liver and skeletal muscle tissues from malin knockout mice. Our data suggest that malin does not target GP for proteasomal degradation. Instead, we found that malin interacted with GP and increased nuclear localization of GP. Malin also altered the glucan‐binding and phosphorylation of GP. Thus, our data show that GP is a novel substrate of malin. Further exploration into the mechanisms of this regulation will unravel insights in glycogen metabolism, physiological functions of the malin‐laforin complex and will aid in betterment of LD pharmacotherapy.Supported by NIH grants R00NS061803, R01NS070899 and University of Kentucky College of Medicine startup funds to MSG and AHA Great Rivers Affiliate Postdoctoral fellowship to VVD.
G protein‐coupled receptor‐1 is an active signaling receptor for the adipokine chemerin
Authors: Rourke Jillian L , Shanmugam Muruganadan , Sinal Christopher J 
Published: June 2021

BACKGROUNDChemerin, is an adipose‐derived signaling protein (adipokine) that regulates adipocyte differentiation and metabolism through activation of chemokine‐like receptor‐1 (CMKLR1). Chemerin also binds to G protein‐coupled receptor 1 (GPR1), suggesting that this protein may be a second chemerin signaling receptor.OBJECTIVETo investigate differential signaling and expression of chemerin receptorsMETHODSReceptor signaling and tissue expression were assessed using a panel of in vitro luciferase‐ reporters and quantitative PCR, respectively.RESULTSGPR1 and CMKLR1 exhibit unique expression profiles which converge with the highest levels of each observed in white adipose tissue (WAT). CMKLR1 is highest in lipid‐filled adipocytes, while GPR1 expression predominates in the WAT stromal vascular fraction. Unlike CMKLR1, chemerin activation of GPR1 does not induce MAPK signaling, but does exhibit higher potency for arrestin recruitment. In common, chemerin activation of both CMKLR1 and GPR1 enhances serum‐response factor (SRF) – mediated gene transcription via a RhoA dependent pathway.CONCLUSIONSThese results demonstrate that GPR1 is an active chemerin receptor and identify RhoA/SRF as a novel chemerin signaling axis. Differential receptor expression suggests that GPR1 may play a distinct role in chemerin function in WAT.Supported by CIHR & Killam Trusts
Localization of fluorescent particles with nanometer precision using standard microscopy and a point spread function approach
Authors: Marki Alex , Ermilov Eugeny , Secomb Timothy W. , Pries Axel R. 
Published: June 2021

BackgroundResolution of optical microscopy is limited by wave optics to about 200 to 400 nm. However, diffraction patterns generated by individual fluorescent particles (point spread functions, PSF) contain location information with much higher precision. Here, we describe a user‐friendly approach for single fluorescence particle 3D localization and tracking with nanometer precision in a standard fluorescence microscope.MethodsImages of fluorescent particles were obtained over a large range (about 7 μm) of vertical positions and analysed by whole pattern (WP) comparison with a PSF recorded for the specific setup and measurement of the outermost ring radius (ORR).ResultsFor a vertical range of 4.5 μm, standard deviation (SD) from the predicted value, indicating validity, was 15.6 and 23.5 nm for WP and ORR. Underexposure or overlapping images led only to a slight increase in SD. The precision, determined as SD of the distance between two particles, ranged from 3 to 6 nm. Local viscosity may be determined from random particle movement with a precision of about 5% of the measured value.ConclusionWith a validity in the range of 20 nm and a precision of about 4–5 nm obtained by standard fluorescent microscopy, the PSF approach offers a valuable tool for a variety of experimental investigations of particle/structure localizations and local viscosity, e.g. in studies of the endothelial surface layer.
Skeletal muscle remodeling in myofiber VEGF‐deficient exercise trained mice
Authors: Delavar Hamid , Rich Benjamin , Wagner Peter D. , Breen Ellen C. 
Published: June 2021

VEGF plays an essential role in regulating angiogenesis and clearance of apoptotic cells. A loss of skeletal myofiber expressed VEGF inhibits exercise‐induced angiogenesis. We hypothesized that skeletal myofiber VEGF deficient mice would increase fibroblast proliferation in hind limb muscles; especially following repeated bouts of exercise.MethodsUsing an HSA‐Cre‐ERT2 X VEGFLoxP (skmVEGF−/−) mouse model, VEGF levels are reduced by >;80% in skeletal muscles of adult mice compared to control littermates (WT). 8 week exercise trained (EX) and untrained (UT) mice were administered BrdU daily during the final week. Proliferating cells (BrdU+) and fibroblasts (vimentin+) were detected by confocal microscopy.ResultsTotal BrdU+ cells/myofiber increased in UT and EX skmVEGF−/− mice compared to the WT‐UT and WT‐EX groups, respectively, in the plantaris (WT‐UT, 6%, skmVEGF−/−UT, 32%, WT‐EX, 37%, skmVEGF−/−EX, 47%) and gastrocnemius (WT‐UT, 2%, skmVEGF−/−UT, 43%, WT‐EX, 25%, skmVEGF−/−EX, 43%). Exercise training, alone, also increased BrdU incorporation in WT muscles. Gastrocnemius sections from exercise trained skmVEGF−/− mice displayed fibrotic foci containing numerous vimentin+/BrdU+ cells. These data suggest that VEGF may prevent muscle injury and fibrosis during exercise training.
Inhibition of Src kinase activity reduces heart rate with slight alteration of heart rate variability
Authors: Jiang Jyun Hao , Ho Jamie , Phan Nhut Nam , Tian Duong Thuy , Do Thi Duyen , Chang Howard , Lin Yen‐Chang 
Published: June 2021

Heart rate variability (HRV) is the cardiac index representing the oscillation of the intervals between consecutive heartbeats (RR intervals in the ECG). It reflects the influences of the autonomic nervous system (ANS) on the sinus node. In order to investigate the involvement of tyrosine phosphorylation in cardiac function of the whole rat system and its effect on HRV, we employed a potent selective inhibitor of Src family kinases, PP2, to inhibit the activity of Src kinases that include Src, Fyn, and Yes.Heart rate was decreased 20% by PP2 at the dose of 0.2 mg/kg. PP3, a negative control compound of PP2, which does not inhibit the activity of Src kinases decreased heart rate by 8% (when at the identical dose of 0.2 mg/kg) when compared with the basal. Interestingly, HRV was not significantly altered at a relatively low dose of PP2 (0.2 mg/kg) when compared with that of PP3 or the basal. Unlike that of PP2, HRV was significantly decreased after application of 0.4 μg/kg isoproterenol. Similar to that of PP2, ZD7288, a specific inhibitor of Hyperpolarized Cyclic‐Nucleotide gated (HCN) pacemaker channels, when at the low dose of 0.2 mg/kg, was able to decrease the heart rate without significant change of HRV in comparison to isoproterenol.Overall, PP2, like ZD7288, can serve as a potential candidate in the treatment of tachycardia‐induced arrhythmias via inhibition of Src‐mediated tyrosine phosphorylation.
Cardiopulmonary Fitness and Cognitive Function in Midlife: Associations with Central Elastic Arterial Stiffness and Regional Cerebral Perfusion
Authors: Tarumi Takashi , Gonzales Mitzi M , Fallow Bennett , Nualnim Nantinee , Tanaka Hirofumi , Haley Andreana P 
Published: June 2021

Central elastic arterial stiffness is associated with accelerated brain aging and cognitive decline. By contrast, habitual aerobic exercise reduces central arterial stiffness and is related to lower risk of dementia in late life.PurposeTo determine the associations among cardiopulmonary fitness, central arterial stiffness, cognitive function, and regional cerebral perfusion in sedentary and endurance‐trained middle‐aged adults.MethodsMaximal oxygen consumption (VO2max), neuropsychological assessment, central arterial stiffness, and regional cerebral perfusion were measured in 27 healthy sedentary (53±1 years) and 32 endurance‐trained (52±1 years) adults.ResultsThe endurance‐trained subjects exhibited greater levels of VO2max, cognitive functions in total composite, memory, and attention‐executive function, and occipito‐parietal blood flow and lower central arterial stiffness than in sedentary subjects (all P<0.05). Carotid distensibility was significantly associated with both VO2max (r=0.28, P=0.046) and total cognitive composite score (r=0.30, P<0.05). Furthermore, central artery stiffness was negatively correlated with occipito‐parietal blood flow (r=−0.41–0.44, P<0.05).ConclusionsLower central arterial stiffness in endurance‐trained adults is associated with greater cognitive function and occipito‐parietal perfusion.
Performing Resistance Exercise Prior to Aerobic Exercise Results in Higher Growth Hormone Levels during Exercise in Physically Active Individuals with Well‐Controlled Type 1 Diabetes
Authors: Yardley Jane Elizabeth , Sigal Ronald J. , Perkins Bruce A. , Riddell Michael C. , Kenny Glen P. 
Published: June 2021

BackgroundPerforming resistance exercise before aerobic exercise (rather than the reverse) can protect against exercise‐induced hypoglycemia in physically active well‐controlled individuals with type 1 diabetes (T1D). Growth hormone (GH) is a potential mediator of this phenomenon as it promotes lipolysis, thereby sparing blood glucose.PurposeThis research aimed to determine whether GH secretion is affected by the order in which resistance (R) and aerobic (A) exercise are performed in people with T1D.MethodsOn separate days, 6 physically active participants with T1D (HbA1c 7.0±1.0%) either ran on a treadmill for 45‐min prior to performing 45‐min of weight lifting (AR) or performed the resistance exercise before running (RA).ResultsGH (mean±SE) rose only transiently in AR, but the rise was larger and more sustained in RA. Differences between sessions were statistically significant for the last 30‐min of exercise (mean at 60 minutes: AR = 2.86±1.03 vs RA = 5.86±1.91 μg/L; at 75 minutes: AR = 1.98±0.78 vs RA = 7.17±2.54 μg/L; at 90 minutes: AR = 1.98±0.82 vs RA = 6.67±2.51 μg/L.ConclusionGH patterns are consistent with this hormone being a mediator in the stabilization of blood glucose levels when aerobic exercise is performed subsequent to resistance exercise in habitually active individuals with T1D. Supported by the Canadian Diabetes Association and the Ottawa Hospital Research Institute.
Gene expression associated with acute mountain sickness in healthy adults rapidly transported to high altitude
Authors: Herman Nicole Marie , Grill Diane E , Anderson Paul J , Miller Andrew D , O'Malley Kathy A , Johnson Jacob B , Richert Maile L Ceridon , Johnson Bruce D 
Published: June 2021

ObjectiveTo identify novel genes and pathways associated with acute mountain sickness (AMS).BackgroundThe uniform, rapid transfer of adults from McMurdo Station (sea level, SL) in Antarctica to the South Pole (SP, 2835m) provides a unique opportunity to study changes in gene expression related to the body's adaptation to hypobaric hypoxia.MethodsVenous blood was drawn at SL and after 2 d at SP in 98 subjects (age 37±9 y, 70% male). Peripheral blood mononuclear cells (PBMCs) were isolated and microarray analysis of the mRNA isolated from these cells was performed (Affymetrix HG U133 Plus 2 microarray chip). Analysis and the computation of expression fold changes and phenotypic regression were performed using the Bioconductor package in R. Pathway analysis was performed using MetaCore™.Results~3200 probe sets with sig. p‐values (p<0.05) and fold changes for each subject were used in a binary logistic regression with the presence or absence of AMS during the 2 d at altitude. Corresponding p‐values and z‐values were entered into MetaCore™ for pathways analysis. Identified pathways included smooth muscle contraction, calcium signaling, and acetylcholine regulation of nerve impulses.ConclusionsProbe sets associated with the presence of AMS had a significant representation in pathways involving aspects of the autonomic nervous system including smooth muscle contraction and nerve signaling. NSF B‐179‐M.