bluuebunny/crossref_metadata_2025_split huggingface dataset:
REAVER: An Improved Image Analysis Pipeline for Quantifying Microvascular Networks
Authors: Corliss Bruce A. , Doty Richard , Matthews Corbin , Peirce Shayn M. 
Published: June 2021

IntroductionAlterations in microvessel networks, including angiogenesis and capillary regression, play key roles in disease, injury, and development. Imaging of microvascular networks can capture their spatial structures, but effective study of network architecture requires methods to accurately quantify them. We present REAVER (Rapid Editable Analysis of Vessel Elements Routine), a tool to analyze blood vessel networks, and demonstrate that REAVER has superior performance compared to the state‐of‐the‐art1–3.Material and MethodsRetinas from C57Bl/6J mice were labeled with IB4 Lectin Alexa Flour 647 and imaged with a Nikon TE‐2000E point scanning confocal microscope. A total of 36 z‐stacks from a variety of tissues were flattened to 2D images with a maximum intensity projection and used as a benchmark dataset. To establish ground truth, all images were manually analyzed in ImageJ. REAVER is written in MATLAB 2018a (Figure 1A).Results and DiscussionREAVER demonstrated markedly higher performance in segmentation and quantification of vessel networks compared to currently available software packages. For vessel length, REAVER had lower mean relative error to ground truth (−1.4% ± 0.091) compared to Angioquant (62.8% ± 0.10), Angiotool (27.0% ± 0.08), and RAVE (43.1% ± 0.17) (Figure 1B). REAVER also had lower error to ground truth with branchpoint count (10.1% ± 0.3) compared to Angioquant (−649% ± 5.06), Angiotool (−341.2% ± 2.90), and RAVE (−1884% ± 12.36) (Figure 1C). Higher accuracy in quantification of vessel networks will allow for improved ability to discern alterations in vessel architecture for microvascular research.Support or Funding InformationFunding: This work was supported by NIH R21 EY028868‐01 and The Hartwell Foundation (to S.M.P.).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ROS Play a Role in Long‐term Gamma Radiation‐induced Heart Damage
Authors: de Oliveira Amanda Almeida , Somasekhar Likitha , Mitra Kunal , Bowles Dawn Elizabeth , Nunes Kenia Pedrosa 
Published: June 2021

Cardiovascular problems are common side effects of cancer therapy, and the heart is one of the major organs that are of concern for potential damage following radiation exposure. Radiation directly affects biological molecules. However, the most impact of radiation‐mediated cell death derives from the generated free radicals, which increase oxidative stress. Reactive oxygen species (ROS) are a hallmark in cardiovascular diseases (CVD), mainly because of its interplay with nitric oxide (NO). NO offers cardioprotection against ROS‐induced damage and ROS, in turn, limits the beneficial effects of NO. Furthermore, radiation induces adaptive alterations in the vasculature leading to vascular dysfunction. Although pathways triggering cardiovascular damage following radiation exposure involve vascular damage in humans, the long‐term mechanisms by which radiation‐induced oxidative stress affects the heart have not yet been clearly defined. We hypothesized that increased ROS contribute to long‐term heart damage following radiation exposure. Flash frozen heart (apex) samples from male C5BL/6J mice (5 months) sacrificed nine months' post radiation was provided by the Department of Surgery at Duke University. Briefly, live animals were subjected to whole body radiation (dose: 200 cGy) at the Brookhaven National Lab. Control animals received Sham treatment. Transverse sections (10 μm) of heart from irradiated and non‐irradiated mice were obtained in a cryostat. Representative images of cardiac fibers were revealed with H&E staining. Additionally, ROS and NO production were evaluated by confocal microscopy. Briefly, the fluorescent probes, Dihydroethidium (DHE) and DAF‐FM Diacetate, were diluted in PBS (5 μM) and the samples were incubated in a humidified chamber at 37° C for 30 minutes. Slides were imaged using a 20x objective and analyzed using ImageJ software (NIH, Bethesda, MD, USA). Protein expression for NOS isoforms was also assessed using a standard Western blotting technique. Our histological analysis indicates loss of cardiac tissue (heart ‐ apex) in irradiated animals compared to Sham animals, suggestive of anatomical remodeling. Confocal microscopy showed an increase in intracellular ROS in irradiated tissue (n=5; p≤0.05), but no significant difference was observed in NO production (n=5; p>0.05). Different NOS isoforms evaluation showed elevated iNOS protein expression levels in irradiated animals. In conclusion, gamma radiation stimulates sustained ROS production in heart tissue and activation of iNOS might be a mechanism for counterbalancing the disruption in the heart antioxidant response system.Support or Funding InformationNASA Florida space Grant Consortium 2018This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Acute Effect of Electronic Hookah Smoking on Endothelial Function in Young Healthy Male Adults
Authors: Ikharo Ebahi , Rezk‐Hanna Mary 
Published: June 2021

Smoking is a major cause of cardiovascular morbidity and mortality worldwide. Over recent years, alternative forms of traditional tobacco smoking have emerged, including electronic (e‐) cigarettes and e‐hookah bowls, with claims that they are safer tobacco alternatives. Unlike e‐cigarettes, e‐hookah bowls are used through traditional water‐pipes, allowing the vapor—containing aerosolized nicotine, propylene glycol, glycerin, and flavorings—to pass through a water‐filled base, potentially altering the vapor, before it is inhaled to the user's mouth. While e‐cigarettes have been shown to cause acute endothelial dysfunction, the effect of e‐hookah is unknown. In eight overtly healthy, young, male adult hookah smokers who do not smoke cigarettes (age 25±3 years; body mass index, BMI 23.9±2.7 kg·m2 mean±SD), we measured heart rate, blood pressure and endothelial function (as measured by brachial artery flow‐mediated dilation (FMD)) before and after a 30‐minute e‐hookah smoking session. Hookah smoking increased heart rate (Δ heart rate: +10±13 beats.min−1) and blood pressure (ΔSBP: +12±7 mm Hg; ΔDBP: +8±5 mm Hg). Importantly, after e‐hookah smoking, FMD decreased from 6.59 ± 2.61 to 5.33 ± 1.97 %Δ, indicating impaired endothelial function. As functional impairment of the endothelium is one of the first recognizable signs of development of atherosclerosis, this new preliminary data provide an initial line of scientific evidence to counter claims that smoking e‐hookah is a safe tobacco alternative. Future studies are indicated to elucidate the long‐term effects of e‐hookah smoking on vascular health, which overtime could accelerate the development of cardiovascular diseases.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Reversal of Endothelial Exosomes‐induced Smooth Muscle Phenotype Transition by 7‐ ketocholesterol Stimulation: Role of NLRP3 Inflammasome Activation
Authors: Yuan Xinxu , Bhat Owais M. , Lohner Hannah , Huang Dandan , Li Pin‐Lan 
Published: June 2021

Recent studies have indicated that endothelial Nod‐like receptor pyrin domain 3 (NLRP3) inflammasome activation during hypercholesterolemia (HC) triggers inflammatory inflammation and consequent atherosclerosis. However, the cell‐cell communication mechanisms between endothelium and smooth muscle cells (SMCs) during inflammation remain poorly understood. The present study hypothesized that activation of coronary endothelial NLRP3 inflammasomes during HC enhances the phenotypic transition of SMCs via exosomes to accelerate atherogenesis. In the primary cultures of carotid endothelial cells (CECs), we found that 7‐ketocholesterol (7‐Ket) treatment activated NLRP3 inflammasome as we showed previously and at the same time it led to a dose‐dependent increase in exosome release as measured by Nanoparticle Tracking Analysis (NTA). The increased exosomes from CECs were found to contain inflammasome products such as IL‐1β, which was associated with the decreased interaction of lysosome with multivesicular bodies (MVBs), as shown by the colocalization of MVB markers VPS16 with IL‐1β or Lamp‐1 under the confocal microscope. Mechanistically, we demonstrated that 7‐keto‐induced exosome secretion with IL‐1β and reduced interaction of lysosomes with MVBs could be enhanced by acid ceramidase (AC) inhibitor‐carmofur, lysosome inhibitor‐bafilomycin A1, or AC gene deletion. When arterial SMCs were incubated with the exosomes collected from CECs with inflammasome activation by 7‐Ket, their phenotypic transition to more synthetic was observed, as shown by increased vimentin or vimentin/SM22 ratio in the Western blot analysis and by enhanced cell migration as measured using wound healing assay method. These effects of purified exosomes from CECs upon NLRP3 inflammasome activation by 7‐Ket were opposite to that induced by exosomes from vehicle‐treated CECs, where decreased vimentin level and cell migration occurred. These results suggest that activation of the NLRP3 inflammasome in CECs by 7‐Ket releases its products via exosome secretion. These exosomes act on SMCs to promote their phenotypic transition to synthetic and also to enhance SMC migration. However, exosomes from CECs without treatment of 7‐Ket control SMC differentiation and reduce their migration, which may maintain normal SMC phenotype and function.Support or Funding InformationSupported by HL057244, HL075316, and DK120491This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hyperglycemia impairs mitochondrial respiration in human brain microvascular endothelial cells
Authors: Chen Allen L. , Sakamuri Siva S. V. P. , Sure Venkata N.. , Evans Wesley R. , Zheng Sufen , Katakam Prasad V.G. 
Published: June 2021

ObjectiveDiabetes is an important risk factor for the stroke. Hyperglycemia‐induced changes in the cerebral vasculature play critical role in adverse effects of diabetes on stroke. Previously, we reported the regulation of vascular function by mitochondria. In this study, we characterized for the first time the effect of hyperglycemia on mitochondrial respiratory function in human brain microvascular endothelial cells (HBMECs) in vitro using the Seahorse XFe24 instrument.MethodsHBVECs were plated on seahorse cell pates and exposed to endothelial cell media containing normal glucose (5 mmol/L), high glucose (25 mmol/L) and normal glucose (5 mmol/L) with mannitol (20 mmol/L) for 24 hours. Basal respiration, ATP production and maximal respiration were measured along with the spare respiratory capacity, proton leak and non‐mitochondrial respiration. Parameters were reported as oxygen consumption rate normalized to the protein concentration of cell lysates in each well (OCR, picomoles/L of O2 per microgram protein). Experiments were repeated three times with 7 to 8 technical replicates for each treatment in each experiment.ResultsBasal respiration was not significantly altered by hyperglycemia, whereas maximal respiration was decreased by 27.5 % when compared to HBMECs treated with normal glucose (7.31±0.32 vs 10.1±0.29, p=0.0002). Spare respiratory capacity was decreased by 47.5% (2.20±0.30 vs 4.25±0.24, p=0.0003) by hyperglycemia, whereas no significant changes were observed in ATP production and non‐mitochondrial respiration. Proton leak showed a strong trend towards decrease (1.09±0.07 vs 1.45±0.03, p=0.06) in high glucose‐treated HBMECs when compared to normal glucose‐treated cells. The observed respiratory changes were due to hyperglycemia but not due to elevated osmotic pressure, as HBMECs treated with mannitol did not exhibit similar respiratory changes.ConclusionsHyperglycemia diminishes the ability of endothelial cells to meet the increased energy demands placed by cellular stress such as neurovascular coupling (physiological) stroke (pathological). Thus, mitochondrial respiratory dysfunction in HBMECs is a novel link between hyperglycemia and cerebrovascular dysfunction.Support or Funding InformationSupport: National Institute of Health: National Institute of General Medical Sciences and National Institute of Neurological Disorders and Stroke (Katakam: R01NS094834).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exposure to WTC Dust Alters Rat Cardiac Gap Junctional Proteins
Authors: Tanwar Vineeta , Bhattacharya Sayak , Rustagi Sachit , Bulgart Hannah , Cohen Mitchell , Wold Loren STATE 
Published: June 2021

ObjectiveFirst responders (FR) present at Ground Zero during the first 72 h after the World Trade Center (WTC) towers collapsed have exhibited increased incidences of cardiovascular disease (CVD) risk. Despite increasing evidences of CVDs, the mechanistic underpinnings behind adverse effects of WTC dust exposure are not well studied. Using a system that generated/delivered supercoarse (10–53 μm) WTC dusts (in a manner that mimicked FR exposures), this study sought to examine potential toxicities in the heart.MethodsIn this study, rats were exposed on two consecutive days for 2 h/day by intratracheal inhalation (ITIH) to supercoarse (10–53 μm) WTC dusts (while under isoflurane [ISO] anesthesia) or an air/ISO (naïve) mixture. This dose conservatively modeled likely exposures to nearly ≈250 mg dust/m3 FR exposure. Hearts were harvested 30 & 60 days post‐exposure and processed for qualitative and quantitative protein expression analyses.ResultsData obtained from right ventricular samples demonstrate decreased desmin expression in the WTC exposed group as compared to the control groups (naive and ISO). Interestingly, although no change in the expression of connexin‐43 was observed between the exposed and control groups, localization studies using confocal microscopy revealed disrupted organization of connexin‐43 at the cardiomyocyte junction. Additionally, confocal imaging suggested reduction of F‐actin expression in the WTC exposed group compared to control groups. Further, expression profiles of the PI3K/Akt signaling axis using western blot analysis revealed a marked decrease in activated phospho‐Akt in the WTC dust exposed group in comparison to control groups. Although blunted activation of Akt was observed, phospho‐PI3K levels were unaltered.ConclusionExposure to WTC dusts caused significant remodeling of gap junctions and altered expression of selected mechanical junction proteins at the intercalated disks. These results illustrate that a defect in a structural/gap junction protein can cause alterations in structural integrity leading to electrical and mechanical perturbations that can contribute to cardiac dysfunction.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evolution of Human Pulmonary Hemodynamics during Severe Sustained Hypoxia
Authors: Hoffmann Fabian , Limper Ulrich , Zaha Vlad , Reuter Hannes , Zange Leonora , Schulz‐Menger Jeannette , Hein Marc , Sadek Hesham , Baldus Stephan , Levine Benjamin , Jordan Jens , Tank Jens 
Published: June 2021

BackgroundSusceptible individuals experience hypoxia‐induced pulmonary hypertension when ascending to high altitude. Descent is considered the primary intervention to treat this condition. A human feasibility study testing influences of severe sustained normobaric hypoxia corresponding to >7,000 m altitude on the heart allowed us to follow the evolution of pulmonary hypertension using state‐of‐the‐art cardiovascular imaging.Material & MethodsTwo healthy, professional mountaineers (subject A, male 57 years, subject B, female 50 years) first pre‐acclimatized for one week in the field at 4559 m altitude. Then, they stayed at the :envihab facility of the German Aerospace Center in Cologne where normobaric hypoxia was achieved by nitrogen dilution starting at 13.5% O2 to a minimum of 8% O2 over three weeks. Afterwards, 8.5% O2 during the daytime and 8.8% O2 at night was kept for another 14 days. Velocity encoded single plane phase contrast MRI of the pulmonary artery (PA) and echocardiography were performed 1 month before, during and 1 and 3 months after hypoxia for assessment of right ventricular (RV) function, PA pressure, area, flow and distensibility.ResultsSystolic PA pressure (PAPsys) was closely related to the decrease in oxygen during acclimatization. In subject B PAPsys peaked at 67 mmHg, 48 mmHg higher than baseline. PAPsys decreased steadily to a minimum of 44 mmHg after two weeks while still at 8.55% oxygen. Peak values occurred with clinical symptoms of acute mountain sickness and a drop in exercise capacity but without signs of cerebral or pulmonary edema. In subject A peak PAPsys was 44 mmHg and remained stable over the entire hypoxia exposure.ConclusionIn healthy humans, severe normobaric hypoxia induces pulmonary hypertension with paradoxically preserved right ventricular function. Pulmonary hypertension may improve over time in some individuals, even in the face of sustained hypoxia. The findings attest to the remarkable physiological reserve of healthy human beings allowing for adaptation to massively increased pulmonary pressure.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The transcriptional regulation of periostin by scleraxis in cardiac myofibroblasts
Authors: Czubryt Michael P , Nagalingam Raghu S , Schwartz Leah Y , Al‐Hattab Danah S , Aroutiounova Nina 
Published: June 2021

Extracellular matrix (ECM) remodeling resulting in cardiac fibrosis is a crucial factor in determining the severity of myocardial dysfunction during heart failure. In response to stress or damage, cardiac fibroblasts undergo phenotype conversion to myofibroblasts, the primary cell type responsible for ECM synthesis and remodeling in the heart. Periostin, a marker of the myofibroblast phenotype, is a matricellular secreted protein that contributes to cardiac remodeling. Periostin plays important roles in mediating cell–matrix signal transduction and in promoting pro‐fibrotic TGF‐β signaling via coupling with matrix associated lysyl oxidase; it is highly expressed in myofibroblasts, but is not expressed in fibroblasts. We have shown that the transcription factor scleraxis induces the phenotype conversion of cardiac fibroblasts to myofibroblasts by directly transactivating the expression of proteins highly up‐regulated in myofibroblasts, including collagen 1α2, fibronectin and α‐smooth muscle actin. Here we report that the treatment of primary adult rat cardiac myofibroblasts with angiotensin II significantly increases periostin gene expression in parallel with increased scleraxis expression. Over‐expression of scleraxis in myofibroblasts increased periostin expression nearly 19‐fold at the mRNA level. TGF‐b treatment induced periostin expression in cardiac myofibroblasts isolated from wild type mice, but failed to show a similar effect in cells isolated from scleraxis null mice. Using in silico analysis, we identified putative E‐box sites within the periostin gene promoter to which scleraxis may bind to potentially regulate promoter transactivation. Our data suggests that scleraxis is necessary and sufficient for TGF‐β mediated periostin expression. Our findings are consistent with a potential role for scleraxis in cardiac fibrosis, where periostin and fibrillar collagen gene expression are elevated in parallel with scleraxis.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The salt‐sensitivity in C57Bl/6J mice is linked to increased renal protein expressions of SLC4A4 and SLC4A5
Authors: Zhu Gangyi , Yu Yanting , Ren Zhiyun , Asico Laureano , Jose Pedro , Wang Xiaoyan 
Published: June 2021

BackgroundRenal regulation of sodium reabsorption largely depends on the function and protein expression of exchanger /transporters /channels for sodium and other ions along the nephron. Decreased protein expression of NCC mediated aldosterone‐escape, a reflection of pressure natriuresis phenomenon. However, the role of renal ion transporters in salt‐sensitive mice is not well understood. Therefore, we studied the effect of varying amounts of sodium intake on the blood pressure (BP) and renal expression of exchanger /transporters/channels for sodium and other ions in salt‐sensitive C57Bl/6J mice and salt‐resistant BALB/c mice.MethodsMale C57BL6 and Balb/c mice were 3 months old, fed normal salt (0.6% NaCl, NS), low salt (<0.02% NaCl, LS) and high salt (4% NaCl, HS) diet, respectively, for 1 week. The mice were placed in metabolic cages for 24 hr urine collection. BP was measured under anesthesia before sacrificing the mice. The kidneys were removed for immunoblotting and immunostaining. Data were analyzed with one‐way ANOVA, Holm‐Sidak test (P<0.05 is significant).Results4% NaCl diet increased the BP (ΔBP=+20 mm Hg) of C57Bl/6J (n=5) but not of sex‐and agematched BALB/c mice (n=5). Additional studies show that C57Bl/6J mice on high NaCl intake (1.6%, 4%, 6%) had higher BP (ΔBP=+20 mm Hg) relative to when they were on low or normal NaCl intake (<0.09%, 0.6%, 0.8%). The dietary salt‐induced increase in BP was observed in anesthetized and conscious, un‐anesthetized mice, measured by telemetry. There were no differences in water/food intake, urinary excretion, and serum concentrations of Na+, K+, and Cl− between the two mouse strains. High salt intake increased the renal protein expressions of SLC4A5 (NBCe2) (162±8% vs. 100±5%) compared to normal salt diet in C57Bl/6J mice, which was not observed in BALB/c mice. High salt intake increased the renal protein expressions of SLC4A4 (NBC1) (163±12%) compared to normal salt diet in C57Bl/6J mice. By contrast, the 4% NaCl diet increased renal SLC26A6 (Pendrin L1) in both C57Bl/6J (190±1%) and BALB/c (161±8%) mice. Increased NaCl intake did not affect the renal protein expression of NHE3 in either mouse strain. Renal protein expressions of NCC and αENaC, but not of NKCC2 and γENaC, were decreased by high salt diet in BALB/c mice (normal response to HS intake), but not in C57Bl/6J mice. SLC4A5 was located in proximal tubules and distal convoluted tubules, colocalized with NHE3 and NCC respectively in the mouse renal cortex. We conclude that the salt‐sensitivity of BP of C57Bl/6J mice is associated with increased renal protein expressions of SLC4A4 and SLC4A5 and impaired salt‐mediated down‐regulation of NCC and αENaC.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.