bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Comprehensive Assay for CFTR Mutational Analysis Using Next‐Generation Sequencing
Authors: Abou Tayoun Ahmad N. , Tunkey Christopher D. , Tafe Laura J. , Levandowsky Elizabeth , Spangler Jessica , Dindinger Matthew , Ross Tristen , Shah Minita , McLaughlin Stephen , Lee Clarence C. , Harkins Timothy T. , Raia Gabrielle , Amos Christopher I. , Wells Wendy A. , Tsongalis Gregory J. 
Published: June 2021

BackgroundCystic fibrosis (CF) is a life‐threatening genetic disorder with over 1000 causative mutations in the CFTR gene detected to date. Current CF genotyping assays target a limited number of mutations and risk missing deleterious ones, both known and novel. Here, we evaluate the performance of a CFTR assay using PCR target enrichment and next‐generation sequencing.Methods68 DNA samples were blindly sequenced on the Ion Torrent Personal Genome Machine (PGM). The CFTR gene was targeted using a custom AmpliSeqTM panel, which consists of 72 amplicons covering all exons and splice sites. Base calls, read alignments, and variant calling were performed using the Torrent SuiteTM software.ResultsBy correctly genotyping 66/68 (97%) of the cases (3/3 normal, 41/42 carriers, 22/23 CF), the first iteration of our assay demonstrates a high sensitivity of calling various pathogenic mutations in CFTR. There was 100% reproducibility of mutation detection between independent runs. The average coverage was greater than 2380 per base pair when 12 samples were pooled on a 318 chip, thus allowing the use of different PGM chips to scale to the desired amount of multiplexing and depth of coverage.ConclusionsThis assay offers clinical laboratories a comprehensive, scalable and cost‐effective means of CFTR genetic testing.
Glycogen storage disease II in Lapland dogs
Authors: Dillard Kati Johanna , Vainio‐Siukola Katri , Vidgren Gabriele , Anttila Marjukka 
Published: June 2021

Glycogen storage diseases (GSD) are rare disorders in animals. Five different types of GSDs have been reported in dogs and cats. They are all due to inborn errors in the function of enzymes involved in glycogen metabolism. GSD type II has been reported in Lapland dog and an autosomal recessive inheritance is suspected. GSD II develops as a result of decreased activity of lysosomal acid α‐glucosidase enzyme. In humans the clinical outcome is variable partly due to the variable amount of remaining activity of the enzyme and the disease has an infantile and late onset form. In Lapland dogs the pathology in GSD II is very similar to the infantile form with the exception of megaesophagus which is present in the affected dog, however, the clinical outcome is different from human.We examined a 1,5 year old female Lapland dog from a litter of three affected dogs. The bitch exhibited progressive muscle weakness and was euthanized due to severe clinical illness. In necropsy markedly enlarged heart and dilated esophagus were present. At the light microscopic level there was accumulation of PAS‐positive material in the cytoplasm and vacuolar degeneration of myocytes within the heart, skeletal muscle and esophagus. Electron microscopy (EM) and measurement of the enzyme activity of the acid α‐glucosidase in blood leucocytes were performed. Further characterization of the cellular pathology is currently under investigation.
The Engraftment Problem: Identifying Proteins that Mediate Stem Cell Adhesion
Authors: Prisco Anthony , Hoffmann Brian R , Kaczorowski Catherine C , Greene Andrew S 
Published: June 2021

Many methods exist to treat heart failure; however no current treatment induces new vessel or cardiomyocyte formation. Recently, researchers have used bone marrow derived endothelial progenitor cells (EPCs) to reverse heart failure in rodents. Unfortunately, results from human studies using similar approaches have been inconsistent. One reason is that only a small percentage of injected cells ultimately engraft into damaged tissue. The first step in engraftment following an IV injection of EPCs is binding of the EPC to the vascular endothelium (VE) within the damaged tissue.Data from our lab has demonstrated that low doses of TNFα (a pro‐inflammatory cytokine) increases the adhesion of EPCs to VE cells in vitro. We hypothesize the protein pairs responsible for mediating the EPC/VE interaction are differentially regulated by TNFα and are located on the surface of the cell. The goal of this study was to identify proteins responsible for mediating EPC/VE adhesion. To identify protein pairs we conducted a novel membrane targeted proteomic based analysis on both EPCs and VE cells treated with TNFα. Candidates were identified based upon stringent, predetermined criteria. Our analysis revealed a high‐confidence list of candidate proteins that are likely responsible for mediating EPC/VE adhesion.
Inhibiton of IRF7 prevents developing corticosteroid‐induced osteonecrosis of the femoral head
Authors: OKAZAKI Shunichiro , NAGOYA Satoshi , KATADA Ryuichi , MIZUO Keisuke , SASAKI Mikito , WATANABE Satoshi , YAMASHITA Toshihiko , MATSUMOTO Hiroshi 
Published: June 2021

ObjectivesWe previously reported that corticosteroid induced osteonecrosis of the femoral head (ONFH) occurs under activation of NF‐ƒÈB and IRF7 by TLR ligand administration, followed by reduction of NF‐ƒÈB activation after corticosteroid treatment. In the present study, we proved that the reduction of IRF7 activity by an IkB kinase inhibitor prevents ONFH.MethodsMale Wistar rats (n=66) were randomly divided into two groups. Each group rats were given ligands of TLR7 on day 1 and methylprednisolone with or without an IkB kinase inhibitor on day 2. Animals were sacrificed 1 or 14 days after last injection. Histopathological and biochemical analyses were then performed.ResultsThe incidence of ONFH was decreased dose dependently by the IkB kinase inhibitor. The activity of IRF7 was significantly decreased after IkB kinase inhibitor administration, but the activity of NF‐kB did not changed.ConclusionsThe present study shows that inhibition of IRF7 activity by the kinase inhibitor prevents ONFH.
Differential inhibition of tissue factor mediated activation of thrombin generation by Dabigatran, Apixaban, Rivaroxaban and Otamaxaban. Pharmacologic implications
Authors: Fareed Jawed , Kahn Daniel , Hoppensteadt Debra , Jeske Walter , Lewis Bruce 
Published: June 2021

IntroductionNewer anticoagulant drugs Dabigatran (D), Rivaroxaban (R), Apixaban (A) and Otamaxaban (O) are monotherapeutic agents targeting thrombin and factor Xa. Beside inhibiting these enzymes these agents also inhibit the generation of thrombin. The purpose of this study was to investigate relative inhibitory effects of D, R, A and O on thrombin generation by using mass spec and functional methods.Materials and MethodsSeveral commercially available prothrombin complex concentrates (PCCs), and normal human plasma (NHP) were supplemented with each of the different inhibitors and proteomic profile was obtained in SELDI‐TOF analysis in the molecular weight (MW) range of 0–150,000. Functional thrombin generation studies were carried out using chromogenic and fleurogenic substrate methods.ResultsAll of the prothrombin complexes exhibited 2 major native peaks at 72 and 13.9 kDa and upon thromboplastin activation showed a 36 and 12.6 kDa peaks representing thrombin generation. D did not inhibit the generation of the 36 and 12.6 peaks whereas O,A and R inhibited the generation of these peaks in a conc dependent manner. Functional generation of thrombin was also inhibited by O, A and R but not by D.ConclusionsThe generation of 36 and 12.6 kDa peak signifies thrombin formation which is strongly inhibited by Xa inhibitors such as O, A and R. D is a relatively weaker inhibitor of this process.
Optimized Caveolin‐1 Scaffolding Domain‐derived peptides for increasing protective nitric oxide production in vascular diseases
Authors: Trane Andy E , Bernatchez Pascal N 
Published: June 2021

Protective vascular nitric oxide (NO), a critical regulator of vascular homeostasis, is produced by endothelial NO synthase (eNOS). Our goal is to promote therapeutic endothelial NO release as a therapeutic avenue for treatment of vascular diseases characterized by decreased NO bio‐availability, which is broadly associated with increased risk of cardiovascular events, such as atherosclerosis. Normally, association of eNOS to Caveolin‐1, the coat protein of caveolae, leads to eNOS inhibition. Using glutathione‐S‐transferase pull down, we identified 10 amino acids from the Cav‐1 Scaffolding Domain (CSD), the section responsible for eNOS inhibition, which binds eNOS. When converted into a cell permeable peptide, we find that the sequence was capable of increasing eNOS dependent NO release from endothelial cells while possessing improved effects on cell morphology compared to full length CSD peptides. In conclusion, we identified a peptide sequence that may serve as the basis for a novel pharmacophore for promoting NO release, providing a new target in cardiovascular diseases, as well as provide insight into caveolae signaling.We would like to thank CIHR, MSFHR and BC Heart and Stroke for funding support.
CDP‐choline protects against sepsis‐induced acute tissue injury in rats.
Authors: Yilmaz Mustafa Sertac , Sevim Cigdem , Altinbas Burcin , Ozyigit Musa Ozgur , Savci Vahide , Yalcin Murat 
Published: June 2021

In the present study, we aimed to determine whether CDP‐choline can improve the outcome of cecal ligation and incision (CLI) in rats. Male Sprague‐Dawley rats were randomly divided into the following four experimental groups: sham, CLI and treated with saline or CDP‐choline. Saline (1 ml/kg) or CDP‐choline (50 and 100 mg/kg) was intravenously injected just after the CLI. Mean arterial pressure (MAP) and heart rate (HR) of all animals were evaluated through the experiment. At 6h after CLI; lung, liver, kidney and spleen tissues were collected. Optical microscopy was performed to examine the pathologic changes. All tissues were stained with hematoxylin‐eosin before examination under light microscopy.In CLI‐animals, MAP decreased significantly throughout observation time compared with sham animals. First significant changes between both groups were observed 3 h after the respective surgical procedure. There was no any significant change observed in HR of the animals. Treatment with CDP‐choline at doses of 50 and 100 mg/kg, i.v. significantly attenuated the CLI‐induced tissue inflammation compared to control animals.In conclusion, intravenously injected CDP‐choline prevents tissue injury induced by CLI. The protective effect of CDP‐choline is not related to its pressor effect.This study was supported by The Commission of Scientific Research Projects of Uludag University, Project number: UAP(T)‐2011/13.
High‐intensity aerobic training restores the ability to modulate sympathetic vasoconstriction in contracting skeletal muscle of hypertensive and healthy middle‐aged individuals
Authors: Mortensen Stefan P , Nyberg Michael , Gliemann Lasse , Thaning Pia , Hellsten Ylva 
Published: June 2021

In young individuals, contracting skeletal muscle can override sympathetic vasoconstrictor activity (functional sympatholysis). To investigate the role of exercise training on functional sympatholysis in hypertension, we measured leg hemodynamics before and after an 8 week period of high intensity exercise training (3–4 times/week) in 8 untrained hypertensive (46±2 years) and 8 healthy controls (47±5 years) during leg exercise and arterial ATP infusion with and without infusion of tyramine. There was no difference in the vasodilatory response to infused ATP between groups. Tyramine induced a similar vasoconstrictor response in both groups and did not alter the vasodilatory response to ATP in either group. Before the training period, exercise hyperemia was lower in the hypertensive subjects (P<0.05) and tyramine lowered exercise hyperemia in both groups (P<0.05). After the training period, exercise hyperemia was similar in both groups and tyramine did not change exercise hyperemia in any of the groups. These results demonstrate that functional sympatholysis is impaired in the leg of both untrained healthy and hypertensive middle‐aged individuals and that exercise training normalizes functional sympatholysis. The vasodilatory and sympatholytic effect of plasma ATP does not appear to be affected by hypertension or the physical activity level.Supported by the Danish Research Council
Sex difference in low dose of angiotensin (ANG) II sensitizing effect on pressor effect of subsequent high dose of ANG II
Authors: Xue Baojian , Zhang Zhongming , Guo Fang , Hay Meredith , Johnson Alan Kim 
Published: June 2021

Previous studies using an Induction‐Delay‐Expression (I‐D‐E) experimental design have demonstrated that one‐week ANG II pre‐treatment can sensitize the brain to produce an enhanced hypertensive response to subsequent ANG II. It has been shown that estrogen can act through the CNS to attenuate ANG II hypertension. The present study was to test whether there is a sex difference in the sensitizing effects of ANG II and the involvement of central estrogen in this sensitization process. Male, intact and ovariectomized (OVX) female rats were implanted for telemetered blood pressure (BP) recording. During I low doses of ANG II alone or concurrent administration with icv estrogen was given for 1 week. After1 week (D) rest, slow pressor doses of ANG II was given for 2 weeks (E).In males and OVX females, ANG II had no sustained effect on BP during I and D. However, during E ANG II‐induced hypertension was greater in the groups receiving ANG II during I in comparison to those groups receiving vehicle. Central administration of estrogen during I blocked low dose of ANG II‐induced sensitization. In intact females, low dose of ANG II actually induced a decrease in BP during I and D. Subsequent high doses of ANG II during E did not result in an enhanced increase in BP in either group. The results indicate that estrogen can play a protective role during the sensitization process and attenuate the development of ANG II‐induced hypertension.
L‐arginine Transporters: A New Treatment Target in Obesity Induced Hypertension?
Authors: Rajapakse Niwanthi W , Karim Florian , Head Geoffrey A , Kaye David M 
Published: June 2021

Cationic amino acid transporter‐1 (CAT‐1) is the predominant L‐arginine transporter expressed in endothelial cells. We hypothesised endothelial CAT‐1 over‐expression can ameliorate obesity induced hypertension.With the use of telemetry, baseline mean arterial pressure (MAP) and responses of MAP to the ganglionic blocker, pentolinium (5 mg/kg) were measured in conscious wild type mice (WT; n = 10) and mice over expressing endothelial CAT1 (CAT+; n=15) fed a normal or a high fat diet for 20 weeks.Both genotypes of mice developed obesity when placed on a high fat diet (Pdiet ≤ 0.007). MAP was greater by 12 ± 2 mmHg (Pdiet < 0.001) in WT mice fed a high fat diet, compared to those fed a normal fat diet. In contrast, MAP was not significantly different in CAT+ mice fed a normal or a high fat diet (Pdiet*genotype= 0.002). Reduction in MAP to pentolinium was greater in WT mice fed a high fat diet (−27 ± 2 mmHg), compared to WT mice fed a normal fat diet (−16 ± 2 mmHg; Pdiet = 0.004) suggesting that the obesity induced hypertension is due to increased activity of the sympathetic nervous system. In CAT+ mice, MAP responses to pentolinium were similar in mice fed a normal or a high fat diet (Pdiet = 0.7; Pdiet*genotype=0.01). These data suggest that endothelial CAT1 over expression can separate obesity from hypertension by buffering the activity of the sympathetic nervous system.
PVN Gαi2 protein‐gated signal transduction ‐ a renal nerve dependent mechanism required for sodium homeostasis and blood pressure regulation in Sprague‐Dawley rats
Authors: Wainford Richard David , Kuwabara Jill Toshiko 
Published: June 2021

AimWe examined the role of PVN Gαi2 proteins, which are upregulated by high salt intake, in maintaining salt‐resistance.MethodsNaïve, bilateral renal denervated (RDNX) or sham RDNX Sprague‐Dawley rats (N=6/gp) received a bilateral PVN infusion of a scrambled (S) or Gαi2 oligodeoxynucleotide (ODN‐300ng/side/day) and a 7‐day normal 0.4% (NS) or high 8% NaCl (HS) diet. On day‐7 MAP, 24h Na+ balance, FENa, plasma norepinephrine (NE), plasma renin activity (PRA), urinary angiotensinsogen (UAGT), and natriuresis to i.v. hydrochlorothiazide (HCTZ‐1.5mg/kg) were assessed.ResultsHS‐intake increased FENa, suppressed plasma NE, PRA, UAGT & natriuresis to HCTZ (P<0.05) without altering MAP in PVN S infused rats. HS intake in intact & sham RDNX rats receiving a PVN Gαi2 infusion evoked elevated plasma NE (NE [nmol/L]; S intact HS 43±4 vs Gαi2 intact HS 108±8, P<0.05), increased natriuresis to HCTZ (peak ΔUNaV to HCTZ [μeq/min]; S intact HS 7.4±0.6 vs Gαi2 intact HS 14.7±0.9, P<0.05), hypertension (MAP [mmHg]; S intact HS 128±2 vs Gαi2 intact HS 147±3, P<0.05), reduced FENa (FENa [%]; S intact HS 0.9±0.1 vs Gαi2 intact HS 0.2±0.1, P<0.05) and suppressed PRA and UAGT. RDNX in HS maintained PVN Gαi2 infused rats prevented hypertension (MAP [mmHg]; Sham RDNX HS Gαi2 149±3 vs RDNX HS Gαi2 132±4, P<0.05), elevated plasma NE, sodium retention and enhanced natriuresis to HCTZ (peak ΔUNaV to HCTZ [μeq/min]; Sham RDNX HS Gαi2 14.8±1 vs RDNX HS Gαi2 8.5±0.7, P<0.05).ConclusionRenal nerve dependent dysregulation of sympathetically driven, but not RAAS mediated, sodium retaining mechanisms, potentially involving altered NCC regulation, occurred following PVN Gαi2‐protein down‐regulation. Therefore PVN Gαi2 protein‐gated pathways represent a central mechanism required to regulate NE mediated sodium homeostasis and maintain salt‐resistance ‐ HL107330.
Early life stress induces altered expression of epigenetic chromatin modification enzymes in aorta and renal vessels
Authors: Ho Dao H. , Loria Analia S. , Pollock Jennifer S. 
Published: June 2021

Early life stress (ELS) is linked to increased risk of adult cardiovascular disease (CVD). Epigenetic pathways are important in cardiovascular maintenance. The current aim is to elucidate whether ELS alters chromatin modification enzyme expression in aorta and renal vessels in adult male Wistar Kyoto rats. Maternal separation (MatSep), a model of ELS, consisted of separating pups from the mother for 3h/day from postnatal day 2 to 14. Unseparated pups served as controls. Gene expression of 84 chromatin remodeling enzymes was assessed in thoracic aorta and isolated renal vessels of 16 week old rats by large‐scale qRT‐PCR array. mRNA expression of histone deacetylase 2 (HDAC2) and protein arginine N‐methyltransferase 2 were significantly decreased in aortas of MatSep rats (0.78±0.05 and 0.76±0.11 fold change relative to control rats (RFC), respectively; p<0.02; n=3/group). MatSep increased renal vessel mRNA expression of genes involved in DNA methylation (DNA methyltransferase 1; RFC=1.2±0.001), and histone phosphorylation (cyclin‐dependent kinase 2; RFC=1.35±0.09), methylation (histone‐lysine‐N‐methyltransferase SUV39H1; RFC=2.49±0.82), and acetylation (HDAC3 and HDAC10; RFC=1.29±0.11 and 1.41±0.06, respectively; p<0.05; n=3/group). Future studies will test whether ELS‐mediated CVD in adulthood is due to derangement of epigenetic pathways in vascular tissue.NIH F32 HL116145; NIH P01 HL69999
SHB deficient mice display an increased GFR and augmented renal arteriolar contractions to both Adenosine and Ang II
Authors: Gao xiang , Sällström Johan , Ma Zufu , Welsh Michael , Persson Erik 
Published: June 2021

BACKGROUNDSrc homology 2 domain‐containing protein B (Shb) is an adapter protein which regulates several signal transduction cascades and endothelial cell functions. Adenosine‐angiotensin II (Ado‐Ang II) interaction plays important roles in regulation of glomerular filtration rate (GFR), vascular resistance and tubuloglomerular feedback. We used Shb‐knockout (Shb−/−) and wild‐type (Shb+/+) mice to investigate GFR and effectiveness of Ado and Ang II to renal resistance vessels.METHODSGFR was measured in conscious Shb−/− and Shb+/+ mice using FITC‐inulin. Isolated and perfused renal afferent arteriolar responses to Ang II doses (10−12 to 10−6M) or low‐dose Ado (10−8 mol/l) alone, as well as Ado (10−8 mol/l) in combination with Ang II doses were studied from both genotypes.RESULTSGFR was increased significantly in Shb−/− (371 ± 12 μL/min) comparing to Shb+/+ (321 ± 11 μL/min). The maximal arteriolar contraction to Ang II was significantly larger in Shb−/− (87 %) than in Shb+/+ (54 %). Low‐dose Ado alone contracted afferent arterioles (6% in Shb−/−; 7% in Shb+/+) and significantly enhanced Ang II arteriolar constrictions (6% in Shb−/− ; 18 % in Shb+/+).CONCLUSIONLow‐dose adenosine augments Ang II arteriolar constriction effectiveness, which indicates Ado‐Ang II interaction in both Shb−/− and Shb+/+ mice. The absence of Shb increases GFR and enhances renal arteriolar contractions to both Ado and Ang II.
Evidence for functional P2X receptors in a mouse cortical collecting duct cell line
Authors: Birch Rebecca , Scott‐Ward Toby , Unwin Robert J , Peppiatt‐Wildman Claire M , Schwiebert Erik , Wildman Scott S P 
Published: June 2021

P2 receptors are potent modulators of key solute and water transport mechanisms in the renal collecting duct (CD) [1]. Controversy exists over the P2 receptor subtype(s) responsible; most studies focus on metabotropic P2Y receptors and arguably ionotropic P2X receptors are largely overlooked.Using an M1 cell line derived from mouse CD and the excised outside‐out configuration of the patch‐clamp technique we have sought functional evidence for extracellular ATP‐gated P2X receptor ion channels. External bath solution contained 145 mM NaCl and the pipette solution contained 145 mM K+ (70 mM Cl−).In outside‐out patches, application of external ATP (100 μM) transiently activated flickery single channel currents (tact ~3s, tdes ~20s). At −100 mV, the mean inward single channel current amplitude (i), open probability (Po) and chord conductance of channels were 0.71±0.12 pA, 0.66±0.12, and 7.7±0.8 pS (n=5), respectively.We demonstrate that ATP directly activates receptors with singlechannel properties similar to those of P2X receptors. Consequently P2X receptors may regulate CD solute and water transport.Research supported by the Wellcome Trust, BBSRC and DiscoveryBiomed.
Renalase protects against cisplatin acute kidney injury in mice
Authors: Desir Gary V , Wang Ling , Velazquez Heino , Moeckel Gilbert , Safirstein Robert 
Published: June 2021

Acute kidney injury (AKI) is an important clinical syndrome for which effective therapies are currently unavailable. Renalase, a novel flavoprotein secreted by the kidney, participates in catechols metabolism protects against ischemic AKI by reducing tubular necrosis, inflammation and apoptosis.The aims of the current study were to investigate the efficacy of renalase in treating cisplatin‐induced AKI, and the mechanisms of its protective action. Three days post treatment with cisplatin (20 mg/kg), plasma creatinine was significantly higher in renalase KO mice compared to WT (1.82±0.56 vs 0.71±0.02 mg/dl, n= 6, P=0.021), as was the renal injury score (Jablonski scale: KO=3.5 vs WT=1.3) indicating that renalase deficiency worsens cisplatin mediated AKI. HK‐2 cells, (human proximal tubular line) exposed to cisplatin for 24 hours showed a 50% decrease in cell viability, and a marked reduction in renalase expression. The addition of exogenous renalase fully restored cellular renalase expression, and increased cell survival. This was associated with decreased BAD, BAX and cleaved caspase 3, and increased BCL2 expression.These data show that renalase protects against cisplatin AKI by decreasing pro‐aopoptotic, and increasing pro‐survival signals. Renalase could provide a useful therapeutic option for cisplatin AKI.