bluuebunny/crossref_metadata_2025_split huggingface dataset:
PTP1B in Calpain Dependent Feedback Regulation of VEGFR2 in Endothelial Cells: Implication in VEGF‐Dependent Angiogenesis and Diabetic Wound Healing
Authors: Zhang Yixuan , Li Qiang , Youn Jiyoun , Cai Hua 
Published: November 2023

The VEGF/VEGFR2/Akt/eNOS/NO pathway is essential to VEGF‐induced angiogenesis. We have previously discovered a novel role of calpain in mediating VEGF‐induced PI3K/AMPK/Akt/eNOS activation through Ezrin. Here, we sought to identify possible feedback regulation of VEGFR2 by calpain via cleavage and activation of its substrate protein phosphotyrosine phosphatase 1B (PTP1B), and the relevance of this pathway to VEGF‐induced angiogenesis, especially in diabetic wound healing. Over‐expression of PTP1B inhibited VEGF‐induced VEGFR2 and Akt phosphorylation in bovine aortic endothelial cells (BAECs), while PTP1B siRNA increased both VEGFR2 and Akt phosphorylation in response to VEGF stimulation. Calpain inhibitor induced VEGFR2 activation can be completely blocked by PTP1B over‐expression. These data implicate calpain/PTP1B feedback regulation of VEGFR2. We next examined a potential role of PTP1B in VEGF‐induced angiogenesis. Endothelial cells transfected with PTP1B siRNA showed faster wound closure in response to VEGF. Aortic discs isolated from PTP1B siRNA transfected mice also had augmented endothelial outgrowth. To further define the role of PTP1B in physiological angiogenesis in vivo, wound healing assay in diabetic mice was carried out. Furthermore, we found that PTP1B over‐expression or calpain inhibition significantly deteriorated wound healing in STZ‐induced diabetic mice. In conclusion, our data for the first time demonstrate a calpain/PTP1B/VEGFR2 negative feedback loop. A critical role of PTP1B as a negative regulator in VEGF‐induced angiogenesis is revealed. Modulation of local calpain and PTP1B activities may prove beneficial in the treatment of defective wound healing in diabetes.Support or Funding InformationThis study has been supported by National Heart, Lung and Blood Institute (HL088975).
A Novel Rat Model of Chronic Thromboembolic Pulmonary Hypertension
Authors: Neves Evandro Neto , Brown Mary Beth , Zaretskaia Maria , Goodwill Adam , Kline Jeffrey 
Published: November 2023

Chronic thromboembolic pulmonary hypertension (CTEPH) pathophysiology is still unclear and deserves more attention. There is a lack of animal models which replicate major pathogenesis of human CTEPH. We hypothesized that the combination of two deleterious insults would be required to damage the pulmonary vasculature and produce sustained pulmonary hypertension and right ventricular (RV) dysfunction found in human CTEPH.CTEPH was induced in Sprague‐Dawley rats (400–450 g) with a combination of microspheres (221.000/100g, 85 μm, i.v.) and VEGF receptor antagonist (SU 5416, 20mg/kg, s.c.) in the PE+SU group. Additional groups received volume‐matched saline (SHAM), microspheres (PE) or SU5416 (SU) separately. CTEPH rats exhibited sustained pulmonary hypertension (RV systolic pressure, 62±12 and 53±9 mmHg at 3 and 6 weeks, respectively) compared to the Sham group (26±2 and 25±2 mmHg). Echocardiography showed robust RV dysfunction, hypertrophy and decreased cardiac output after CTEPH (P<0.05). Similarly, impaired RV contractile function (developed pressure and dP/dtmax) was observed by Langendorff's technique and was accompanied by increased RV mass in PE+SU group (>2 fold compared to Sham group, P<0.05). Approximately 25% decrement in aerobic capacity was observed in rats in PE and PE+SU groups after 3 days post‐injection. While more robust decreases were found after three (40%) and six (32%) weeks in PE+SU group, rats in PE group had their aerobic capacity recovered (P<0.05). Abundant lung vascularization can be observed in the PE group, whereas lung vascular rarefaction along with elevated endothelial injury makers were found in PE+SU group.We conclude that at least two injuries are required to replicate pulmonary obstruction and microvasculature injury to reproduce CTEPH in rats. Interestingly, progressive PH, RV dysfunction and marked hypertrophy were found associated with deficient lung vascularization in our rat model.Support or Funding InformationAHA 14POST20170013 (E Neves PI). This study was made possible by the Lilly Endowment, Inc. Physician Scientist (J Kline PI).
The Blurry Vision Project
Authors: Biro Mark P , Brantley Thomas , Freeman Barbara 
Published: November 2023

Background/ObjectiveNeuroanatomy students traditionally learn the visual pathway, along with other neurologic pathways, in a two‐dimensional format. This approach is widely used but not thoroughly tested against other methods of learning. The experimental objective was to use the visual pathway as a platform to compare learning methods using a two‐dimensional approach versus a novel three‐dimensional model.MethodsThirty‐four students from the neuroanatomy course offered at the Case Western Reserve School of Medicine were recruited for the study. Subjects represented a number of graduate programs including the medical school, masters of anatomy, speech language pathology, biomedical engineering and neuroscience doctorate program. These students were separated into control and experimental groups prior to the experiment. The control group consisted of 13 students and the experimental group had 21 students. Each group completed a similar lesson plan reviewing components of the visual pathway and visual deficits secondary to lesions in the pathway. The control group used pencil and paper to reconstruct and explore the visual pathway, while the experimental group followed along with a three‐dimensional model of the visual pathway. Students in each experimental limb worked in groups of three to four students and given forty minutes to complete the first portion of the experiment. Upon completion, students were provided 20 minutes to individually take a quiz composed of fifteen multiple‐choice questions and a five question Likert scale questionnaire regarding their experience.DataAll data was compared using two‐tailed t‐tests for independent samples and significance determined by p‐value < .05. Objective data from the experiment indicated students in experimental limb had higher mean scores (10.71) than control students (8.92) and outperformed them on twelve of fifteen total questions (all but 5,9 & 12), but these values only approached significance. Subjectively students from the experimental group had higher mean values for Likert scale questionairres (21.14 vs 19.84) and higher means on all individual questions, but similarly values only approached significance. Additionally, experimental groups had a 105% attendance rate, whereas the control group had 62% attendance rate.ConclusionsFrom the experiment it cannot be concluded that students preformed significantly better in the three‐dimensional construct group compared to the two‐dimensional learning approach, despite higher mean subjective and objective data. This may in part due to overrepresentation of the experimental group and limited power of the experiment. Future experiments with a larger student population and additional subjective and objective questions will be necessary to determine if this novel three‐dimensional model approach to neuroanatomy education is superior to traditional approaches.Support or Funding InformationSupport for the experiment was provided by the Case Western Reserve University School of Medicine.
A Novel Model of Obstructive Sleep Apnea for Studying Cardiovascular Disease
Authors: Heintz Timothy , Lux Jacob , McLerie Meredith , Durgan David J , Panama Brian K 
Published: November 2023

BackgroundObstructive sleep apnea (OSA) is associated with cardiovascular disease. OSA is characterized by transient upper airway closures during sleep (apneas), and up to 24% of adults in the United States have OSA, many of whom are unaware of their condition. Patients with OSA exhibit daytime sleepiness, may have mood disorders, and are at increased risk for arrhythmias, such as a trial fibrillation. Despite the association between OSA and cardiac disease the specific mechanisms remain elusive. We have initiated a recently developed novel rat model of OSA which closely recapitulates the human condition by incorporating true apneas. Our goal was to examine the effects of OSA on several genes associated with cardiac fibrosis.Methods and ResultsTo generate our OSA model, a silicone balloon was surgically implanted in the trachea of a rat (aged 45–70 days). To produce apneas, the balloon was inflated to transiently obstruct the airway. Unanesthetized, freely‐ranging rats were randomly given sixty 10‐second obstructions (apneas) per hour during their sleep phase (09:00–17:00) for 14 days. Control rats had obstruction devices implanted but the balloons were not inflated. Balloon inflations caused decreases in O2 saturations in OSA rats from 95.6±0.2% to 74.8±1.2%, and there were no differences in O2 saturations when balloons were deflated. O2 saturation decreases caused by apneas were also consistent throughout the experimental period. Measurements of mRNA from the ventricles after two weeks of OSA showed 2‐fold increases in the inflammatory cytokines TNF‐α and TGFβ‐2.ConclusionsOSA is associated with increased cardiac expression of genes implicated in cardiac pathology. Our data demonstrates the utility of this novel rat model to study OSA‐induced cardiac disease.
Maternal High‐fat Diet and Diabetes Impairs Cardiac Metabolism and Mitochondrial Dynamics in Adult Offspring of Diabetic Pregnancy
Authors: Mdaki Kennedy S , Larsen Tricia D , Louwagie Eli J , Baack Michelle L 
Published: November 2023

BackgroundOffspring of diabetic mothers are at risk of cardiovascular disease at birth and throughout life. We recently reported that maternal high‐fat (HF) diet impairs cardiac function in newborn offspring of diabetic pregnancy through metabolic stress and mitochondrial dysfunction. Whether this in utero exposure to maternal hyperglycemia and dyslipidemia can have lasting consequences on the developing heart is still unknown.ObjectiveTo determine the long‐term effects of prenatal exposure to maternal diabetes and high‐fat (HF) diet on cardiac metabolism in the adult offspring.MethodsFemale Sprague Dawley rats received controlled (CD) or HF diet throughout the study. At gestational day 14, dams were injected with either citrate buffer (CB) placebo or streptozotocin (STZ) to induce diabetes producing offspring from the following groups: controls (CD‐CB), diabetic exposed (CD‐STZ), HF exposed (HF‐CB), and combination exposed (HF‐STZ). Litters were culled to equal size and pups were cross‐fostered to normal dams. Cellular bioenergetics, oxidative injury, and mitochondrial dynamics were evaluated using Seahorse® XF24 analyses, malondialdehyde assay, and confocal live cell imaging of primary isolated cardiomyocytes from offspring at 6 months of age. Outcomes were interrogated for diet, diabetes and interaction effect using ANOVA with significance set at p≤0.05.ResultsHF diet‐exposed cardiomyocytes demonstrated abnormal cardiac bioenergetics profiles, increased lipid peroxidation, and a relatively decreased ability to recover from FCCP challenge with signs of mitochondrial dysfunction on live cell imaging. Combination‐exposed cardiomyocytes were most severely affected.ConclusionLate gestation diabetes alters cardiac metabolism in the developing heart and the effects are exacerbated with prenatal exposure to a maternal HF diet. These cardiometabolic consequences persist into adulthood. The FCCP challenge demonstrates that combination exposed offspring have an impaired ability to maintain ATP production under stress which is a significant risk factor for heart disease. Together, our findings highlight that cardiac metabolism plays a significant role in developmentally programmed cardiovascular disease in offspring of diabetic pregnancy.Support or Funding InformationNIH‐NICHD K08HD078504, Sanford Research, SSOM‐USD Faculty Grant, Sanford Health– SDSU Collaborative Research Seed Grant, Institutional Development Awards (IDeA) NIH ‐GMS P20GM103620‐01A1 (Pediatrics) and P20GM103548 (Cancer) COBRE grants
Elementary TRPV4 sparklets in the endothelium dilate mouse pulmonary arteries
Authors: Marziano Corina , Hong Kwangseok , Cope Eric L , Sonkusare Swapnil K 
Published: November 2023

Endothelial dysfunction is a hallmark of pulmonary disorders including pulmonary hypertension, pulmonary edema, acute lung injury and ischemia. Ca2+ influx through TRPV4 (transient receptor potential vanilloid 4) channels plays an important role in regulating the function of pulmonary endothelium, and abnormal TRPV4 channel activity has been associated with lung injury, ischemia, and pulmonary edema. However, the function of TRPV4 channels and their regulation by physiological stimuli in the pulmonary endothelium remain unknown. The objectives of this study were to elucidate the regulation of TRPV4 channels in pulmonary endothelium under physiological conditions; and to identify the Ca2+‐sensitive targets that mediate TRPV4‐induced vasodilation in these arteries. Third or higher order pulmonary arteries (PAs) were isolated, slit‐open and pinned down, and the Ca2+ signals were recorded using a spinning disk confocal imaging system. In the presence of cyclopiazonic acid to deplete the internal stores of Ca2+, TRPV4 channel function was recorded as Ca2+ influx events‐TRPV4 sparklets (Sonkusare et al., Science, 2012). The activity of TRPV4 sparklets, estimated as fluorescence integral over a field of view, was increased by 3‐ and 17‐fold in the presence of 3 and 10 nM GSK1016790A (TRPV4 agonist), respectively, when compared to the baseline. The TRPV4 sparklets were inhibited by TRPV4 channel inhibitor HC067047 (1 μM); and were absent in the presence of 0 mM extracellular Ca2+ in both the endothelium‐denuded arteries and the arteries from TRPV4 knockout mice. A multi‐Gaussian curve fit to the all‐points histogram of the fractional fluorescence traces revealed a single channel amplitude of 0.18 ΔF/F0 in the mice that express Ca2+ biosensor GCaMP2 selectively in the endothelium, and 0.28 ΔF/F0 in the arteries loaded with fluo‐4AM. The TRPV4 sparklet amplitudes were not affected by a change in the agonist concentration, or in the presence of TRPV4 inhibitor. The effect of TRPV4 channel activation on arterial diameter was studied in the PAs cannulated and pressurized to 10–20 mm Hg and pre‐constricted with U46619 (50–100 nM). GSK101 (30 nM) induced a 67% vasodilation which was mostly inhibited by NG‐nitro‐L‐Arginine (nitric oxide synthase (NOS) inhibitor, 100 μM). Our results support the concept that TRPV4 sparklets in PA endothelium represent single TRPV4 channel openings that cause vasodilation via NOS activation. These results will enable direct studies of TRPV4 channel regulation in PAs under healthy and diseased conditions.Support or Funding InformationSupported by NIH award HL121484 to SKS
Resting Measures of Cardiovascular Autonomic Control are not Influenced by Energy Drink Ingestion or Postural Changes in Healthy Adults
Authors: Leatherwood Matt D , Herron Robert L , Carter Stephen J 
Published: November 2023

INTRODUCTIONEnergy drink consumption is common among young people and linked to acute alterations in cardiovascular‐autonomic profiles. However, the magnitude and relevance of such changes is equivocal. This study evaluated resting autonomic control, as evidenced by heart rate variability (HRV), during postural changes following ingestion of a commercially‐available energy drink (ED) and zero‐kcal control drink (CD).METHODSUsing a single‐blind, repeated measures, counter‐balanced crossover design, 14 (7 women) young (mean ± SD; 25 ± 3 y), healthy adults completed two testing sessions after an overnight fast. Participants were asked to avoid vigorous exercise and caffeine consumption the day prior to testing. Each session was separated by at least 48 h. Baseline HRV measures were recorded in the supine (Sup) and standing (St) positions prior to ingesting either an ED or control drink (CD) of equal volume. Following a 30‐min digestion period, HRV measures were repeated. The alternate drink treatment was consumed during the subsequent visit. The change in pre‐ to post‐drink HRV recordings were calculated and analyzed via repeated‐measures, two‐way ANOVA.RESULTSANOVA results (Drink x Body Position) did not detect mean differences in HRV time‐ or frequency‐domains of interest (Interaction and main effects p > 0.05, RMSSD: SupED = −0.11 ± 27.13 ms, SupCD = −2.59 ± 27.18 ms, StED = 14.64 ± 15.84 ms, StCD = 10.23 ± 16.34 ms; HF nu: SupED = −1.59 ± 14.87 nu, SupCD = 5.67 ± 16.62 nu, StED = 12.02 ± 15.69 nu, StCD = 6.36 ± 18.50 nu; LF/HF: SupED = 0.06 ± 0.66 nu, SupCD = −0.14 ± 0.74 nu, StED = −2.36 ± 4.45 nu, StCD = −2.40 ± 9.41 nu).CONCLUSIONAccording to these data, consumption of a commercially available ED does not alter cardiovascular autonomic control in young, healthy adults under resting conditions.
Suitability of Evasin‐3 as Molecular Imaging Marker for Early Coronary Neovascularization by Arteriogenesis
Authors: van Lier Monique GJTB , Schwarz Janina CV , de Vos Judith , de Bruin Cora M , Verberne Hein , Siebes Maria 
Published: November 2023

AimTo assess labeled molecular targets as early imaging biomarker for formation of coronary collaterals in ischemic rabbit hearts. Evasin‐3 binds specifically to CXCL1 and can provide a powerful molecular agent to visualize early arteriogenesis.MethodsWe induced regional myocardial ischemia in 10 New Zealand White rabbits by ameroid constriction of a major coronary artery. Animals were sacrificed 9 or 16 days after surgery. Fluorescently labeled (Alexa‐Fluor 488) evasin‐3 was injected by cardiac puncture and circulated for 1.5 or 3hrs prior to sacrifice in 5 rabbits. Coronary arteries of 5 excised hearts were filled with UV Blue fluorescent replica material. Using an imaging cryomicrotome, alternate cutting (14 μm slice thickness) and multispectral episcopic imaging (14 μm resolution) yielded detailed 3D representations of the heart and vasculature co‐registered with the labeled molecular marker. Vascular distribution of the fluorescent imaging biomarker was determined.ResultsFluorescence cryomicrotome imaging clearly depicted evasin‐3‐AF488 in coronary vessels. The overlay with vascular cast images revealed presence of evasin3 along all major vessels. However, no enhanced uptake was visualized in the ischemic area.ConclusionsThe distribution of fluorescently labeled evasin‐3 in coronary vessels was not restricted to vessels in the ischemic area. It is therefore not well suited as a specific imaging marker of neovascularization by arteriogenesis.Support or Funding InformationThis research was supported by the Center for Translational Molecular Medicine (EMINENCE).
Cardiovascular risk profile in Peruvian medical students
Authors: Fernández‐Rodríguez Lissett Jeanette , Bardales‐Zuta Victor Hugo , Avalos‐Alvarado Carlos Eduardo , Hilario‐Vargas Julio Santos 
Published: November 2023

We report the results of a cross‐sectional study that queries cardiovascular risk in second year medical students enrolled at Antenor Orrego Private University, Trujillo, Peru. We selected 78 students (58 women, 20 men) randomly. At the beginning of the semester, we determined medical and family history, age, adverse habits, somatometrics, blood pressure, baseline blood glucose, lipid profile and electrocardiograms of the participants over one week. The average age was 18.90 ± 1.5 for women and 18.70 ± 1.2 men. 39.7% of women and 50% men came from Trujillo, 58.6 and 75% had a family history of diabetes mellitus or arterial hypertension, 55.2 and 45% had medical history, 5.2 and 10% smoke, 16 and 35% had systolic blood pressure ≥ 130 mmHg and 7 and 20% had diastolic pressure ≥ 85 mmHg. The 38 and 55% were overweight, 3% and 10% were in obesity grade I, 2% and 5% in grade II and 5% males in grade III. The waist ‐hip index for 71% of women was > 0.80 and was > 0.95 for 30% of men. 2 and 10% had high triglycerides. 9% of women presented total cholesterol at the upper limit. 69% and 50% had low HDL, 3 and 2% of women had high and very high LDL. 46.6% had electrocardiographic abnormalities. We found that cardiovascular risk factors such as changes in lipid metabolism and ECG, overweight and obesity were common among medical students. This study underlines that universities should implement primary prevention measures to reverse or slow this trend among late‐teenage students, as earlier intervention can improve lifespan and outcome in the long term.Support or Funding InformationAntenor Orrego Private University.
β‐ENaC associated with cilia indicates the distribution of absorptive epithelial cells in the lumen of small airways
Authors: Flores‐Delgado Guillermo , Quinton Paul Marquis 
Published: November 2023

Airway surface liquid (ASL) is crucial for optimal clearance of mucus, pathogens and debris from the pulmonary bronchial tree and is managed by a balance between secretory and absorptive activities along the luminal airway surfaces. Immunohistochemically, we recently demonstrated that in small airways, about half of the lining epithelium consists of cells with a secretory phenotype that are confined to the contraluminal regions (pleats) of the plicated luminal epithelium (1). However, the distribution/location of absorptive airway epithelial cells has not been demonstrated. The epithelial sodium channel (ENaC) is responsible for Na(+) absorption across the apical membranes of many salt absorbing epithelia including airways. Herein, we hypothesized that localization of β‐ENaC subunits would define the distribution of absorptive cells in airway epithelia.We obtained small pieces of peripheral porcine lung tissue of approximately 0.5 cm3 and pieces of trachea of approximately 0.5 cm2 that were dissected and immediately snap‐frozen at −80°C. Serial tissue sections (7 μm thick) containing cross‐sections of small airways, with a diameter between 0.5 and 1 mm, were used for immunohistochemistry. We used antibodies directed to b‐subunit of ENaC (courtesy C. Fuller), as well antibodies directed to β‐Tubulin and the tight junction protein, ZO1. Western blot analysis demonstrated ENaC expression in cell extracts derived from isolated small airways and trachea. Confocal microscopy of immunostained sections of airway tissue defined the distribution of β‐ENaC subunits in the apical regions of a sub‐set of cells in the small airways and trachea. In small airways, strong staining for β‐subunit was generally restricted to ciliated cells in the region around the tips of folds in the luminal epithelium well separated from the sub‐sets of secretory cells in base of the epithelial pleats where staining was generally absent or faint, but strong for the secretory marker, NKCC1, seen previously (1). In trachea, β‐ENaC subunit staining was mainly observed at the surface of the stratified ciliated epithelium, and absent in regions where submucosal glands were present. Importantly, we observed that β‐subunit of ENaC appeared to co‐localize with the distribution β‐tubulin, a biomarker for cilia.These findings suggest that small airway epithelium contains a subset of epithelial cells with absorptive phenotype, recognized by the high expression of β‐subunit of ENaC.Support or Funding InformationSupported by the Cystic Fibrosis Foundation‐MCCC, CFRI and the Nancy Olmsted Trust for Pediatric Respiratory Medicine