bluuebunny/crossref_metadata_2025_split huggingface dataset:
Proteomic Characterization of Epicardial‐Myocardial Signaling Reveals Novel Regulatory Networks Including a Role for NF‐κB in Epicardial EMT
Authors: Li Yanyang , Urban Alexander , Midura Devin , Simon Hans‐Georg , Wang Q. Tian 
Published: November 2023

Reciprocal signaling between the epicardium and underlying myocardium not only is essential for heart development, but also plays a critical role during heart regeneration in regeneration‐competent vertebrates. The complex molecular interactions and regulatory networks involved in this communication are not well understood. In this study, we integrated mass spectrometry with bioinformatics to systematically characterize the secretome of embryonic chicken EPDC‐heart explant (EHE) co‐culture. The 150‐protein secretome dataset established here greatly expands the knowledge base of the molecular players involved in epicardial‐myocardial signaling. We identified proteins and pathways that are implicated in epicardial‐myocardial signaling for the first time, as well as new components of pathways that are known to regulate the crosstalk between epicardium and myocardium. The large size of the dataset enabled bioinformatics analysis to deduce functional networks for the regulation of specific biological processes and predicted signal transduction nodes within the networks. We performed functional analysis on one of the predicted nodes, NF‐κB, and demonstrate that NF‐κB activation is an essential step in TGFβ2/PDGFBB‐induced epithelial‐to‐mesenchymal transition. In summary, we have generated a global perspective of epicardial‐myocardial signaling for the first time, and our findings open exciting new avenues for investigating the molecular basis of heart development and regeneration.Support or Funding InformationThis work was funded by the Chicago Biomedical Consortium with support from the Searle Funds at The Chicago Community Trust.
Pravastatin&[trade] Prevents Venous Shear Stress Dependent Induction of Aquaporin 1 Protein Expression in Human Umbilical Vein Endothelial Cells in vitro
Authors: Crum Raphael J. , Krane Carissa 
Published: November 2023

BackgroundAquaporins (AQPs) are transmembrane water channels that facilitate osmotically‐driven transmembrane water flux. AQP1 is abundantly expressed in vascular endothelium, where it has been shown to function in endothelial cell migration, wound healing and cell volume regulation in response to mechanical stimuli. AQP1 protein abundance is enhanced in cultured human umbilical vein endothelial cells (HUVECs) in response to fluid flow shear stress. Statins, a pharmacological class of HMG‐CoA reductase inhibitors, have been shown to suppress the early development of vein graft intimal hyperplasia by improving endothelial cell function, likely by affecting endothelial response to arterial shear stress. Statins have also been shown to differentially regulate the expression of specific AQPs by altering their abundance and/or subcellular localization. Therefore, it is hypothesized that AQP1 may function as a component of the mechanosensory complex in endothelial cells, and is subject to shear‐stress induced gene regulation and modulation of expression by statins.MethodsHUVECs were cultured in microfluidic chambers under static (0 dynes/cm2) and venous fluid shear stress (6 dynes/cm2) in the presence or absence of 5 μM Pravastatin™. Fluorescent immunocytochemistry was used to detect AQP1 protein expression and subcellular localization. Images were captured using confocal laser scanning microscopy. Mean fluorescent intensity (in arbitrary units) of AQP1 immunofluorescence under each condition was determined using ImageJ software, and normalized to control conditions (time zero, static culture with no Pravastatin™). 25–30 cells from two independent experiments were analyzed for each experimental condition.ResultsAQP1 immunofluorescence increased 1.3‐fold (p<0.05) after 24 hours, and 1.7‐fold (p<0.05) after 48 hours in cells cultured under venous fluid shear stress as compared to the control. This shear stress‐induced expression was blocked by incubation with Pravastatin™ (p>0.05; control vs. venous shear stress with Pravastatin™). Pravastatin™ had no effect on AQP1 immunofluorescence in static cultures (p>0.05 vs. control). These data indicate that Pravastatin™ prevents shear stress dependent induction of AQP1 in cultured HUVECs.ConclusionsThe results presented show that the shear stress dependent induction of AQP1 protein expression is prevented by Pravastatin™. These results further demonstrate a possible tissue‐independent function of statin drugs in regulating the expression of AQPs in general and specifically a potential cardioprotective benefit of statins, independent of their cholesterol‐lowering function, related to the regulation of AQP1 in vascular endothelial cells.Support or Funding InformationThis research was funded by the Schuellein Chair in the Biological Sciences (CMK), University of Dayton Honors Program (RJC), and a 2015 UGSURF Award to RJC.
Bronchodilator Therapy and Survival in Heart Failure Patients
Authors: Uithoven Katelyn E , Snyder Eric M , Olson Thomas P 
Published: November 2023

INTRODUCTIONBronchodilator therapy via β‐agonists has been considered a controversial treatment option for patients with cardiovascular disease, including heart failure (HF), due to previous studies suggesting increased rates of mortality. A major limitation to several previous studies is the lack of similar baseline characteristics between groups; therefore, a matched control comparison is yet to be explored.PURPOSETo evaluate the relationship between bronchodilator therapy and survival in HF patients.METHODSA retrospective analysis was performed on 96 HF patients who underwent a cardiopulmonary exercise test between the years 1994–2010. Patients were divided into two groups: 48 patients on bronchodilator therapy (BDT) and 48 matched controls (CTL). Patients were matched based on age (yrs), height (cm), weight (kg), body mass index (BMI), New York Heart Association (NYHA) classification, and cardioselective β‐blocker medication during the time of their cardiopulmonary exercise test. Patients were followed for an average of ten years for an endpoint of cardiovascular mortality. An independent samples t‐test was performed comparing BDT and CTL for age (yrs), height (cm), weight (kg), and BMI, to determine comparable baseline characteristics. A Chi‐Squared test compared NYHA classification, cardioselective β‐blocker status, and sex between BDT and CTL. A log rank test was performed to determine differences in survival distributions between BDT and CTL.RESULTSThere were no significant demographic differences between groups (age: 59.8 vs. 60.3yrs; height: 172.1 vs. 171.9cm; weight: 86.9 vs. 86.2kg; and BMI: 28.9 vs. 28.6kg/m2 for BDT vs. CTL, respectively). NYHA classification (χ2=0.00; p=1.0), cardioselective β‐blocker status (χ2=0.14; p=0.71) and sex (χ2=0.202; p=0.65) were not different between groups. BDT patients demonstrated an event rate of 51.1% compared to 42.6% for CTL patients over the same time period. The survival distribution for BDT compared to CTL was not statistically different, χ2(1) = 1.49, p=0.22.CONCLUSIONThese data suggest that HF patients on bronchodilator therapy demonstrate no significant difference in survival compared to matched HF controls. In contrast to previous work, our data suggests bronchodilator therapy does not elicit increased mortality in these HF patients.
A Regulatory Role for SDF‐1/CXCR4 Axis In Promoting TNF‐mediated Cardiac Damage Associated With Alcoholic Cardiomyopath
Authors: Little Janay , Abad Adam , Peeran Ahmad , Haddad George , Tarzami Sima 
Published: November 2023

INTRODUCTIONAlcoholic cardiomyopathy is a form of heart disease caused by alcohol abuse. Tumor necrosis factor alpha (TNF‐α) plays a role in cardiomyopathy, and can lead to cardiomyocyte apoptosis. Our preliminary data suggests that stromal cell‐derived factor 1 (SDF‐1) stimulates TNF‐α mRNA and protein secretion in cardiomyocytes. Since cardiac expression of SDF‐1 is enhanced in inflammatory cardiomyopathy, we have suggested a regulatory role for SDF‐1/CXCR4 axis in promoting TNF‐α mediated cardiac damage associated with cardiomyopathy.HYPOTHESISOur central hypothesis is that increased levels of SDF‐1 correlate directly with production of TNF‐α and the severity of pathological changes observed in the setting of alcoholic cardiomyopathy. Vascular endothelial growth factor (VEGF) is linked to vasculogenesis, which is seen in healthy hearts. The decrease in quantity of VEGF has been seen as a contributor to damage of heart tissues along with the increase in SDF‐1 and TNF‐α; studies have presented this to be true, but has yet to be shown or seen in alcoholic hearts.METHODSLittermate adult rats were put on a 3‐months isocaloric Lieber‐Decarli liquid diet with high alcohol (HA: 100mM) levels. Hearts were taken out and concentrations of TNF‐α, SDF‐1, and VEGF were analyzed in protein lysates that were prepared from those hearts. TNF‐α, SDF‐1, and VEGF protein expression was quantified using ELISA assay. To evaluate whether SDF‐1 upregulation mediated TNF‐α expression and subsequent cell death, adult rat cardiomyocytes were freshly isolated and treated with SDF‐1 at different concentrations and apoptosis was assessed using TUNEL at 24hrs post treatment.RESULTSOur data suggests that high alcohol consumption is associated with reduced cardiac contractile function. Moreover, in the setting of alcoholic cardiomyopathy, our preliminary results suggests that SDF‐1 can stimulate production of TNF‐α and subsequent apoptosis possibly contributing to the observed pathophysiology. The ELISA reported that the amount of TNF‐α and SDF‐1 increased in the alcoholic tissues compared to the controls; however, there is a trend toward VEGF downregulation in the alcoholic tissues compared to the controls which can further exacerbate the severity of damage. Treatment of isolated adult rat cardiac myocyte with SDF‐1 at higher concentrations (>30nM) upregualted TNF‐α protein expression and induced subsequent apoptosis.CONCLUSIONOur data suggests a SDF‐1/CXCR4 network regulates TNF‐α production in cardiomyocytes leading to pathological changes observed in alcoholic cardiomyopathy e.g. myocyte apoptosis.Support or Funding InformationResearch report in this publication was supported by the American Physiological Society under grant number NHLBI; R25 HL115473‐01
Assessment of cardiac function during the progression of heart failure induced by intracoronary embolizations in the canine model: monitoring in conscious and anesthetized states.
Authors: Krishnan Abhinav C , Lovelace Abe , Senador Danielle , Kaur Jasdeep , Alvarez Alberto , Hanna Hanna W , Levanovich Peter E , Dombrowski Mary D , Mannozzi Joseph T , Aung Kimberly , Haddad Samuel , Sala‐Mercado Javier , Levy Phillip D , O'Leary Donal S 
Published: November 2023

Various animal models of acute and chronic heart failure exist including coronary artery occlusion via ligation or microembolization, rapid‐ventricular pacing, aortic banding, and fluid challenge. However, to our knowledge, no study has investigated whether anesthesia alters conclusions regarding hemodynamic effects associated with heart failure development. In this study, global ischemic cardiomyopathy, via repeated coronary artery microembolizations, (range: 5–15 embolization procedures per animal) was induced to lower ejection fraction to < 40% in ten chronically instrumented canines. We measured the progression of myocardial dysfunction over time, with each animal serving as its own control in conscious and anesthetized states to elucidate the effect of anesthesia on cardiovascular parameters. Hemodynamic measurements were taken in three conditions: anesthetized, immediately prior to each embolization, anesthetized 1.25 minutes after each embolization, and in a conscious, non‐anesthetized state at rest, 1 week after each embolization. We observed that anesthesia lowered arterial pressure (Δ−31.8 ±3.7%) and cardiac output (Δ−22.2 ±3.4%) when compared with the conscious state and that the effect of the embolization on ventricular function was better revealed in conscious animals. Thus, anesthesia affects the conclusions drawn regarding the extent of cardiovascular impairment induced via the embolization model.Support or Funding InformationSupported by Novartis Pharmaceuticals CRLX030AUSNC01T and NIH HL55473 and HL126706.
The Roles of Flanking DNA and Transcriptional Activators in Regulating SAGA‐Mediated Nucleosome Acetylation
Authors: Olson Sannie J. , Mittal Chitvan , Shogren‐Knaak Michael A. 
Published: November 2023

The SAGA family of transcriptional coactivators are prototypical nucleosome acetyltransferase complexes that regulate multiple steps of gene transcription. The size and complexity of both the SAGA enzyme and the chromatin substrate provide opportunities to regulate the acetylation process. To better probe this regulation, we have characterized the binding interactions and kinetics of acetylation using different nucleosomal substrates and the full SAGA complex purified from budding yeast. We find that SAGA‐mediated nucleosome acetylation proceeds through a multiple‐turnover burst phase, suggesting rapid nucleosome acetylation at multiple sites prior to nucleosome release. The rate of nucleosome acetylation is affected by DNA flanking the nucleosome, both by facilitating the binding of SAGA and by accelerating acetylation turnover. This stimulation requires flanking DNA on both sides of the nucleosome, where one side needs to be longer than 15 base pairs. Gal4‐VP16 activator can also augment nucleosome acetylation. However, contrary to our expectations, this stimulation does not appear to occur by stabilizing the binding of SAGA toward nucleosomes containing an activator binding site. Instead, stimulation of nucleosome acetylation occurs by increasing acetylation turnover by SAGA. Altogether, these studies uncover several novel mechanisms of SAGA regulation by chromatin substrates.Support or Funding InformationThis work was supported by American Cancer Society Research Scholar Grant 1206501 to M. A. S.‐K. Work by S. J. O. was supported in part by a Gary Roewe Faculty Research Award.
Mitofilin knockdown in Rat H9c2 Myoblasts Induces Cell Death by Apoptosis Via an AIF‐PARP‐Dependent Mechanism and Cell Cycle Arrest
Authors: Lie Mihaela , Madungwe Ngonidzashe Benson , Feng Yansheng , Bopassa Jean C. 
Published: November 2023

IntroductionMitochondrial activity plays an essential role in the efficient function of cardiomyocytes. Mitofilin is an inner membrane protein that has been defined as a mitochondria‐shaping protein in controlling and maintaining mitochondrial cristae structure and remodeling. In this study, we determined the role of mitofilin in the H9c2 myoblast survival by investigating the mechanism underlying mitofilin knockdown‐induced cell death by apoptosis.MethodsH9c2 cells were cultured and treated with mitofilin siRNA or scramble siRNA for 24 hours. Cell death (apoptosis), caspase 3 activity, and cell cycle phases were assessed by flow‐cytometry, cytochrome C release and intracellular ATP production were measured by ELISA. Mitofilin, AIF and PARP expression was measured by Western blot analysis and calpain activity assessed using calpain kit. Mitochondria images were taken using electron microscopy.ResultsWe found that mitofilin knockdown increases cell death by apoptosis mainly via activation of AIF pathway leading to nuclear fragmentation and subsequent PARP1 activation in the nucleus that is correlated with S phase arrest of the cell cycle. Knockdown of mitofilin in H9c2 myoblasts with siRNA led to mitochondrial swelling and damage of mitochondrial cristae that is associated with the increase in ROS production, decrease in intracellular ATP production and a marked decrease in mitochondrial membrane potential. Moreover, cells treated with mitofilin siRNA displayed an increase in calpain activity versus scramble siRNA.ConclusionTogether, these results indicate that mitofilin knockdown by siRNA increases calpain activity that presumably leads to mitochondrial structural degradation resulting in a critical reduction of mitochondrial function that is responsible for the increase in cell death by apoptosis via AIF‐PARP mechanism which is associated with nuclear fragmentation and S phase arrest of cell cycle.Support or Funding InformationVoelcker Fund and American Heart Association.
Vaping to Vascular Damage: The Role of E‐Cigarettes on Vascular Function
Authors: Clayton Stuart , DeVallance Evan , Branyan Kayla , Pitzer Christopher , Breit Matthew , Hoskinson Hannah , Klinkhachorn Powsiri , Erdreich Brett , Chantler Paul , Olfert Mark 
Published: November 2023

BackgroundElectronic cigarettes (E‐cigs) have exponentially increased in popularity and usage due to the perception that e‐cigs are safe, their usefulness as a cigarette‐smoking cessation tool, and the appeal of heavily marketed flavors. There is little known about the long‐term effects of E‐cig vapor exposure, particularly in the context of vascular dysfunction. We hypothesize that the long‐term use of E‐cig vapor decreased aortic function, and increase aortic stiffness.MethodsData were obtained from C57BL/6 female mice exposed to 3R4F reference cigarette (N=5–7), cappuccino flavored E‐vapor (18 mg/ml nicotine, N=7), or filtered air (N=7–8) for 4 h/day, 5 d/wk for 8 months. Food and water were administered ad libitum. Before and after the exposure, in‐vivo aortic stiffness (pulse wave velocity) was measured using B‐mode and Doppler ultrasound by obtain blood flow signals at the aortic arch and before the carotid bifurcation from a single image, which were gated to the EKG. At the end of the exposure the thoracic aorta was dissected, sectioned into rings and mounted onto an ex‐vivo wire tension myograph system. Force transduction was used to measure the changes in aortic tension in response to methacholine, or sodium nitroprusside.ResultsAortic stiffness increased (0.45±0.20 m/s) in the air‐exposed group, reflecting the normal aging process. However, an accelerated age‐associated aortic stiffness was noted in the cigarette (1.28±0.27 m/s) and E‐cig (1.14±0.24 m/s) groups (ANOVA, p<0.05). In animals exposed to filtered air, the maximal aortic relaxation achieved to methacholine was 90%, compared to 60% and 70% in the cigarette and E‐cig groups, respectively (p<0.05). No differences were noted in sodium nitroprusside dilation between groups.ConclusionOur data suggests that 8 months of E‐cig significantly accelerated the age‐associated increase in aortic stiffness, and significantly impaired aortic endothelial‐dependent but not endothelial‐independent dilation. These data show the E‐cigs induce similar vascular dysfunction to cigarette smoke exposure suggesting that E‐cig have similar risk to develop accelerated cardiovascular aging and disease.Support or Funding InformationNational Institutes of Health Grant: 1P20 GM‐109098West Virginia University – Marshall University Collaborative Health Research Grants
Critical role of smooth muscle Rac1 in neointimal formation following vascular injury
Authors: Lennon Frances E , Li Yuxin , Hong Zhigang , Liao James K 
Published: November 2023

BackgroundRac1 is a small GTPase, which plays a critical role in cell movement, spreading, proliferation, transformation, and production of reactive oxygen species (ROS). Because smooth muscle cell (SMC) migration and proliferation are important for vascular remodeling after injury, we hypothesized that SMC Rac1 may play an important role in neointima formation.Methods and ResultsTo investigate the role of SMC Rac1 in neointimal formation, we used global haploinsufficient Rac1 knock out mice (Rac1+/−) and also generated SMC‐specific Rac1+/− (smRac1+/−) mice by crossing smMHC‐Cre mice with Rac1 flox/flox mice. Four weeks after carotid artery ligation, neointimal formation in Rac1+/− and smRac1+/− mice was substantially reduced compared to wild‐type (WT) or Rac1 flox/+ (control) mice (intima area, WT vs. Rac1+/−: 59.59±12.43 vs. 27.72±5.97 103 μm2; control vs. smRac1+/−: 50.69±9.75 vs. 14.32±2.43 103 μm2). Aortic SMC isolated from smRac1+/− mice exhibited normal cytoskeletal architecture, and similar migratory and adherent function as WT SMCs. However, smRac1+/− SMCs showed decreased chemotaxis, and proliferation compared with WT SMCs. Furthermore, SMCs from smRac1+/− mice showed decreased phosphorylation of PAK1 and ERK, and reduced ROS production in response to mechanical stretch compared to that of SMCs from WT mice. Inhibition of Rac1 in vitro using NSC23766 lead to decreases in vascular SMC migration and proliferation and significantly reduced SMC glycolytic metabolism measured via changes in ECAR and lactate production. The effects of Rac1 inhibition on migration and proliferation could be rescued via supplementation with the glycolytic end products lactate or pyruvate.ConclusionThese findings indicate that Rac1 regulation of SMC migration and proliferation is mediated in part via control of glycolytic metabolism in these cells. These findings illustrate the critical role of Rac1 and glycolysis in neointimal formation after vascular injury. Therapeutic modalities, which target Rac1 in SMCs, therefore, may be beneficial in preventing vascular proliferative diseases.Support or Funding InformationProject funded by NIH R01 HL052233 to James K Liao
Perinatal Taurine Supplementation Prevents Metabolic and Cardiovascular Effects of Maternal Dyslipidemia in Adult Rat Offspring
Authors: Thaeomor Atcharaporn , Tangnoi Chonticha , Chaisakul Janeyuth , Seanthaweesuk Suphaket , Somparn Nuntiya , Roysommuti Sanya 
Published: November 2023

Maternal obesity predisposes metabolic and cardiovascular disorders in adult offspring. This study tests the hypothesis that perinatal taurine supplementation prevents dyslipidemia and hypertension in adult offspring of dyslipidemic rat mothers. Female Wistar rats were fed normal rat chow and water with (Dyslipidemia group) or without dyslipidemic induction by intraperitoneal Triton WR‐1339 injection (Control group). After confirmed pregnancy, they were supplemented with (Control+T and Dyslipidemia+T) or without 3% taurine in water from conception to weaning. After weaning, male offspring were fed normal rat chow and water throughout the study. At 16 weeks of age, body weight, epididymis fat size, fasting blood glucose (Control, 84 ± 2 mg/dl; Control+T, 85 ± 3 mg/dl; Dyslipidemia, 96 ± 2 mg/dl; Dyslipidemia+T, 82 ± 2 mg/dl; P < 0.05), plasma low‐density lipoprotein (Control, 102 ± 6 mg/dl; Control+T, 100 ± 6 mg/dl; Dyslipidemia, 137 ± 7 mg/dl; Dyslipidemia+T, 107 ± 5 mg/dl; P < 0.05) and cholesterol (Control, 81 ± 5 mg/dl; Control+T, 81 ± 3 mg/dl; Dyslipidemia, 95 ± 3 mg/dl; Dyslipidemia+T, 81 ± 2 mg/dl; P < 0.05), mean arterial pressure, and heart rate increased but baroreflex sensitivity decreased significantly in Dyslipidemia compared to Control groups. While perinatal taurine supplementation did not affect these parameters in the Control, this treatment normalized, at least in part, the metabolic and cardiovascular effects of maternal dyslipidemia in adult offspring. In addition, plasma high‐density lipoprotein, triglyceride, leptin, alanine aminotransferase, aspartate aminotransferase, creatinine, and blood urea nitrogen were not significantly different among the four groups, irrespective of maternal dyslipidemia or taurine supplementation. These data demonstrate that perinatal taurine supplementation prevents the adverse effects of maternal dyslipidemia on metabolic and cardiovascular defects in adult male offspring.Support or Funding InformationSuranaree University of Technology