bluuebunny/crossref_metadata_2025_split huggingface dataset:
VEGFR2 signals with VE‐cadherin and PECAM to facilitate leukocyte transendothelial migration
Authors: Sullivan David P. , Fu Tao A. , Weber Evan W. , Gonzalez Annette P. , Dalal Prarthana J. , Rutledge Nakisha S. , Muller William A. 
Published: May 2022

The migration of leukocytes out of the blood and into tissue is a critical component of the inflammatory process. To gain access to the target tissue and exert their beneficial or deleterious effects, leukocytes undergo a complex series of adhesive interactions with endothelial cells, which line the vessel wall. In the putative committed step in this process, the leukocyte must find the junction between adjacent endothelial cells and crawl through. This process, commonly referred to as transendothelial migration (TEM), has long been known to require homophilic interaction between PECAM on leukocyte and the endothelial cells. Recently our group showed that the process also involves calcium signaling originating through the cation channel TRPC6.We hypothesized that PECAM may be functioning in TEM using some of the interactions recently shown to be involved in its role in the mechanotransduction of fluid sheer stress. Perhaps engagement of leukocyte PECAM with endothelial cell PECAM at the cell border provides the sheer stress in this case. Here we report the unexpected role of a macromolecular PECAM complex that facilitates signaling to TRPC6 in TEM. Specifically, we show that endothelial cell PECAM binds to VEGFR2 and VE‐cadherin via the transmembrane domain of VE‐cadherin. Swapping the VE‐cadherin transmembrane domain with that of N‐cadherin abrogated TEM. Likewise, deletion of the cytoplasmic c‐terminal tail of PECAM also abolished this signaling cascade. Stimulation of this complex with antibodies against PECAM induces the phosphorylation of VEGFR2 on tyrosine Y1175. Mutation of that critical residue to phenylalanine (Y1175F) prevented VEGFR2 phosphorylation and blocked TEM. Furthermore we show that this complex signals specifically to PLC1γ (and not PLCβ3) leading to its phosphorylation. Blocking its function with the inhibitor U73122 also blocked TEM. Diacylglycerol liberated by PLCγ1 activates TRPC6.We also confirmed the role of VEGFR2 in vivo using to separate models of inflammation. First, we examined the recruitment of neutrophils to inflamed skin using the croton oil dermatitis model. In control animals, neutrophils are robustly recruited to and migrate across post‐capillary venules. In VEGFR2 knockout (KO) mice, most neutrophils are found inside the vessel with a corresponding decrease in those found out in tissue. Similarly, live animal imaging (intravital microscopy) of the inflamed cremaster muscle showed no defect in neutrophil rolling or adhesion in the VEGFR2 KO animals, but a profound disruption in their ability to undergo TEM compared to control wild type mice.Taken together, these data confirm the role of a VEGFR2/PECAM/VE‐cadherin mechanotransduction complex that signals through PLC1γ to facilitate leukocyte TEM upstream of the action of TRPC6. Uncovering this signaling pathway opens up several new targets for potential therapeutic intervention.
Examining Vascular Responses to Passive Movement in Premenopausal Females: Comparisons Across Sex and Menstrual Cycle Phase
Authors: Weggen Jennifer B. , Hogwood Austin C. , Decker Kevin P. , Darling Ashley M. , Chiu Alex , Richardson Jacob , Garten Ryan S. 
Published: May 2022

PurposeVascular responses to passive movement represent a novel assessment of vascular function and serves as a critical component for adequate blood flow regulation during exercise. Although alterations to estradiol (E2) throughout the menstrual cycle, as well as sex differences, are known to impact vascular function, limited research exists in determining whether vascular responses to passive movement are altered in premenopausal females when compared across sex or menstrual cycle phase. We hypothesized that there would be no differences in leg blood flow responses to passive leg movement (PLM) in premenopausal females when compared to age‐matched males or when evaluated across follicular menstrual cycle phases, when E2 fluctuates the most.MethodsVascular responses to one‐minute of PLM were assessed in eleven premenopausal females (n = 21; 23 ± 4 yrs) and age‐matched males (n = 21; 24 ± 4 yrs). All the females completed the PLM assessment during the early (EF) and late follicular (LF) phase of their menstrual cycle, based on self‐reporting of menses. Lower limb microvascular function was assessed during PLM via changes in leg blood flow measured as peak change from baseline (LBFΔPEAK) and area‐under‐the‐curve (LBF AUC). Thigh volume, a known modulator of PLM responses, was also assessed in the passively moved limb in all participants via anthropometric measurements. Blood samples were obtained at rest from the antecubital vein of the arm to determine differences in E2 between the EF and LF phases of the menstrual cycle.ResultsThigh volume differed significantly between the females and age‐matched males (F: 4.9 ± 1.5 L; M: 8.2 ± 1.6 L; p < 0.01) and, when examined in all participants, thigh volume was significantly correlated to PLM‐induced hyperemia evaluated as both LBFΔPEAK (r = 0.6; p < 0.01) and LBF AUC (r = 0.4; p = 0.014). When the group differences in thigh volume were accounted for in the PLM analysis by employing thigh volume as a co‐variate, no significant differences in LBFΔPEAK (F: 542 ± 444; M: 794 ± 444 ml; p = 0.15) and LBF AUC (F: 164 ± 227; M: 249 ± 227 ml; p = 0.3) were revealed. The females reported no significant differences in PLM‐induced blood flow responses across menstrual cycle phases (EF vs LF) evaluated as LBFΔPEAK (EF: 385 ± 186; LF: 386 ± 154 ml; p = 0.9) or LBF AUC (EF: 123 ± 100; LF: 58 ± 84 ml; p = 0.11). Plasma E2 levels were significantly higher in females in the LF phase (215 ± 149 pg/ml) when compared to the EF phase (149 ± 98 pg/ml) of the menstrual cycle (p = 0.01).ConclusionThis study revealed no differences in PLM‐induced LBF responses in premenopausal females when compared to age‐matched males or when assessed across follicular menstrual cycle phases.
Exercise Training Alleviates Alzheimer’s Disease‐Associated Aortic Endothelial Dysfunction
Authors: Lee Junghoon , Zarezadehmehrizi Aliasghar , Hong Junyoung , Lee Jonghae , Park Yoonjung 
Published: May 2022

BackgroundsA strong association between cardiovascular disease and Alzheimer’s disease (AD) has been reported, but its possible mechanism has been unknown. We have tested whether aortic endothelial function in AD is impaired and investigated the protective effect of exercise training (EX) on endothelial function in aorta of AD mice.METHODSWild type (C57BL/6, WT) and AD (APP/PS1dE9, AD) mice underwent treadmill training (WT‐EX and AD‐EX) or remained sedentary for 12 weeks. The expression of total‐and phosphorylated‐endothelial nitric oxide synthase (t‐eNOS and p‐eNOS), beta‐amyloid, and superoxide dismutase 2 (SOD2) were quantified by Western blot analysis.RESULTSt‐eNOS and p‐eNOS expressions in AD were decreased compared to WT but EX increased both t‐eNOS and p‐eNOS in AD (P < 0.01 and P = 0.09). Beta‐amyloid levels in AD were higher compared to WT but EX decreased it (P < 0.05). SOD2 levels in AD were increased by EX (P < 0.05) whereas there was no change in WT.CONCLUSIONRegular aerobic exercise alleviates e‐NOS‐mediated endothelial dysfunction in aorta of AD mice possibly via reduced beta‐amyloid and increased SOD2 levels.
Genesis of Antibiotic Resistance (AR) LXXXVI: Turbulence Modeling(TM) of Hemodynamics in Simplified Severe Sepsis Protocol‐2 (SSSP‐2)‐NCT01663701: Mechanism(s) of ISP Afloat Transient APG Thrust VKVS free shear flow augmented cerebral vessel collapse, GCS3, & IHM
Authors: Montoya Hector , Flores Jorge , Kannan Subburaj 
Published: May 2022

Based on the theoretical inferences from the CFD pertaining to VKVS/vortices/shedding frequency/vortex‐induced vibration/ hysteresis effect, Shed Vortex, and Karman/symmetric vortex shedding, Courtesy: Flow‐3D HydroÒ, COMSOL Multiphysics®, © 2021, NASA‐U.S: https://earthobservatory.nasa.gov/images/85989/canary‐islands‐kick‐up‐von‐karman‐vortices, Harvard Natural Sciences Lecture Demonstrations http://sciencedemonstrations.fas.harv… https://www.youtube.com/watch?v=_Hbbkd2d3H8, Vortex shedding; https://www.youtube.com/watch?v=ahohd8ceGR4, and hemodynamic data from NCT01663701, here we present a plausible physiological basis for the effect of fluid bolus augmented VKVS, eliciting ISP, thrust skewed viscosity/velocity profile rupturing and cause the cerebral hemorrhage, that leads to precipitous ihm in the usual care cohort of SSSP‐2 (PMID: 28973227). Hypothesis:Fluid bolus augmented VKVS augment incompressible free shear flows in cerebral arteries ensue decomposition of glycocalyx/endothelium as a function of time, viscosity and velocity profiles. Geometry:RBC, WBC, platelets, p47; p53; Table 2.1 p63, Table 2.2, p64; Fig 2.15 p 78; ISBN:9780128024089. Governing Equation, Derivation and Prospects (Physiological Connotation): Subsequent to fluid bolus induced fluid volume expansion is a function altered viscosity (hematocrit) dependent velocity profile as a function of time augment five different free shear flow: a. far wake, downstream of ZoO, b. mixing layer, downstream of ZoO occurring between two parallel streams with different velocity profile, c. plane jet, d. round jet, occur downstream of ZoO, and e. constrained radial jet, occur when two jets of similar velocity profile, (possibly different viscosity) in a turbulent flow; Assumptions based on the hemodynamic data from NCT016637001: I. Aforesaid free shear flow has self‐similarity, self‐preserving; Velocity component: Equation, 3.22; II. two velocity profiles located downstream at locations x1 and x2 are not same: equation, U (x, y)/uo(x) [or (U‐U (x, y)/U0(x) for the wake); III. Boundary layer approximations are not valid for aforesaid free shear flows, IV. molecular transport momentum differs at different location; V: pressure not the same thought vessels, VI: no continuity (equation of motion refer equation 3.23; 3.24; VII: closure coefficient differs for different points in the cerebral/ coronary circulation, applicable governing equations for the eddy‐viscosity model, Eq:3.25, p60‐61; ISBN‐13: 978‐1928729082. Taken together, fluid bolus induced fluid volume expansion augment cyclical anaerobic/aerobic milieu conferring altered generation time(G): G=t/n, where time (t) per generation (n = number of generations) for ClPr exacerbating toxic shock expedite irreversible loss of vasculature penultimate ihm in usual care group of NCT01663701.
Effects of different types of resistance exercise failure training on the methylation status of genes that drive skeletal muscle hypertrophy
Authors: McIntosh Mason , Sexton Casey , Godwin Josh , Ruple Bradley , Osburn Shelby , Hollingsworth Blake , Agostinelli Philip , Kavazis Andreas , Ziegenfuss Timothy , Lopez Hector , Smith Ryan , Young Kaelin , Dwaraka Varun , Mobley Christopher , Sharples Adam , Roberts Michael 
Published: May 2022

ObjectiveWe sought to determine how one bout of resistance training to failure with either higher repetitions (30FAIL) or lower repetitions (80FAIL) affected the promoter methylation statuses of genes that drive skeletal muscle hypertrophy.HypothesesWe hypothesized that a bout of 80FAIL training would lead to a more robust hypomethylation of genes that regulate skeletal muscle hypertrophy compared to 30FAIL training.MethodsEleven previously‐trained college‐aged men (age: 23 ± 4 years, 11.42 ± 6.38 percent fat, 4 ± 3 years training experience) volunteered for this study. Each participant underwent two training bouts (spaced one week apart) involving either: i) 30FAIL training; 4 sets of back squats and 4 sets of leg extensors to failure at 30% of one‐repetition maximum (1RM), or: ii) 80FAIL training; 4 sets of both exercises at 80% of 1RM. Muscle biopsies from the vastus lateralis were collected prior to each bout (PRE), 3 hours following each bout (3hPOST), and 6 hours following each bout (6hPOST). Following the conclusion of the study, tissue was batch‐processed for DNA isolation, and DNA was subjected to the Illumina MethylationEPIC array. In an a priori fashion, genes that have been shown to induce skeletal muscle hypertrophy in genetic mouse models were the target of this investigation (Verbrugge et al. Frontiers Physiol, 2019).ResultsTotal training volume (sets x reps x load) between the 30FAIL and 80FAIL bouts were not significantly different (p= 0.571). Differentially methylated region changes for the following genes from PRE to 3hPOST and PRE to 6hPOST are presented herein: SKI, FST, AKT1, ACVR2B, MSTN, KLF10, RHEB, IGF1, PAPPA, PPARD, IKBKB, FSTL3, ATGR1, UCN3, MCU, JUNB, NCOR1, GPRASP1, GRB10, MMP9, DGKZ, PPARGC1A, SMAD4, LTBP4, BMPR1A, CRTC2, XIAP, DGAT1, THRA, ADRB2, ASB15, CAST, EIF2B5, BDKRB2, TPT1, NR3C1, NR4A1, GNAS, PLD1, CRYM, CAMKK1, YAP1, INHBA, TP53INP2, INHBB, NOL3, and ESR1. Additionally, significant differences between conditions at each time point are highlighted.ConclusionsThis study continues to display how different modalities of resistance training affect the skeletal muscle molecular milieu and furthers our scientific understanding of factors that contribute to training adaptations.
RBM20 regulates muscle hypertrophy through titin and calcium signaling
Authors: Gao‐Hu Jeffrey S. , Guo Wei 
Published: May 2022

Study objectiveRNA binding motif 20 (RBM20), an RNA binding protein, regulates alternative splicing of titin and calcium handling genes that mediate hypertrophy signaling. The objective of this study is to investigate whether RBM20 regulates muscle hypertrophy through titin and/or calcium signaling.MethodsSoleus (SOL) and extensor digitorum longus (EDL) were collected from wild‐type (WT) and Rbm20 knockout (KO) rats. Transcriptome profiling was analyzed by RNA sequencing (RNA‐seq). Hypertrophic signaling was examined using HeLa cells as well as differentiated and undifferentiated C2C12 myoblasts. RBM20 was knocked down in C2C12 myoblasts by siRNA transfection. Both immunostaining and western blotting were performed to test expression of hypertrophy associated proteins in both cell lines.ResultsRBM20 KO promoted SOL hypertrophy and EDL atrophy in rats. Hypertrophy‐associated hallmark proteins CaN, Erk and p38 were elevated in KO SOL, but not in KO EDL, while atrophy‐associated hallmark protein p62 was increased in KO EDL, but not in KO SOL. RNA‐seq data identified differentially expressed and spliced genes associated with hypertrophy in both EDL and SOL. Titin was switched to largest isoform as a result of alternative splicing in both KO SOL and EDL muscles. To test whether titin is the major mediator that regulatew hypertrophic signaling in RBM20 KO rats, we transfected Rbm20 gene into HeLa cells that do not express titin. The results revealed that the levels of hallmark proteins did not changed with overexpression of RBM20. To further test the role of titin in RBM20‐mediated hypertrophy, Rbm20 was knocked down in undifferentiated C2C12 cells which do not express titin as well, and no change was observed for hallmark proteins. C2C12 myoblasts were then differentiated to myotubes and reduced RBM20 in differentiated C2C12 cells promoted reduced expression of hypertrophic hallmark proteins.ConclusionOur results suggest that RBM20 regulates muscle hypertrophy in a muscle type‐dependent manner, and RBM20‐mediated hypertrophy is regulated via titin, a hypertrophic mediator.
The Impact of the COVID Pandemic on STEM Professionals
Authors: Benore Marilee A. , Esquivel Amanda O. , Marincean Simona 
Published: May 2022

Early reports indicated that the COVID 19 pandemic negatively impacted STEM professionals' research, resulting in fewer grants, patents, and publications, and disrupted student learning, especially in skills development. More concerning was the prediction that the most severe consequences would be felt by young STEM professionals, or those with family care responsibilities. In order to investigate the impact of the COVID pandemic on STEM professionals in the USA we conducted a survey. Questions included loss of research and lab time, data on papers, grants and publications, and the disruption to personal obligations and work life balance.The survey, conducted about one year into the pandemic, provided evidence that there were significant differences in productivity and responsibilities, with some differences and work load aligned with gender. Response comments reflected both negative and positive outcomes. This data is consistent with trends observed in other professional fields, as well as those reported in other nations.This poster will present the results of this research, along with outcomes from other studies. The concerns have spurred several calls to action by professional societies, government agencies and other groups. These recommendations attempt to ensure that STEM professionals can continue to progress in their careers. Strategic planning is necessary in order to ensure that the next generation of STEM researchers are properly supported in order to ensure a diverse workforce.
Polybromo‐1 missense mutations found in renal cancer patients affect bromodomain stability and biological function
Authors: Jiao Guanming , Goetz Christopher , Bursch Karina , Nuñez Raymundo , Olp Michael , Zimmerman Michael , Urrutia Raul , Smith Brian , Dykhuizen Emily 
Published: May 2022

Chromatin structure is important in modulating gene expression. A central component of chromatin structure is the covalent post‐translational modification (PTM) of histones. Distinct histone PTMs can determine the accessibility of chromatin and recruit different effector proteins and components of the transcriptional machinery. Histone are modified by “writer” and “eraser” enzymes that can add and remove PTMs, which are recognized by evolutionarily conserved reader domains. Bromodomain is one type of evolutionarily conserved “reader” domain composed of ~110 amino acids, including a conserved asparagine that specifically binds acetylated lysine. There are approximately 80 bromodomains in humans, primarily in chromatin‐associated proteins. Most proteins contain one or two bromodomains. The exception is Polybromo‐1, which contains six tandem bromodomains in addition to two bromo‐adjacent homolog (BAH) domains involved in protein‐protein interactions and a high mobility group (HMG) for interacting with DNA. PBRM1 is a subunit of the polybromo‐associated BAF (PBAF) chromatin remodeling complex, one of three major SWI/SNF subcomplexes found humans. These chromatin remodeling complexes alter chromatin accessibility by evicting, sliding and exchanging nucleosomes on DNA.In clear cell renal cell cancer carcinoma (ccRCC), the most common subtype of kidney cancer, PBRM1is the second most mutated gene after VHL, with PBRM1mutations found in approximately 40% of ccRCC patients. Mouse models indicate that PBRM1 deletion paired with VHL deletion in the kidney results in tumors, establishing PBRM1 as a bona fide tumor suppressor. While most PBRM1 mutations in patients are also loss of function mutations, ~15‐20% of PBRM1 mutations are missense mutations, with a large portion found at highly conserved residues in the fourth bromodomain (BD4). Using recombinant PBRM1 BD4 mutants, we have assessed how the most common missense mutations found in patients affect the stability and acetyllysine binding of PBRM1 BD4, revealing new insight into bromodomain structure and function. In addition we have expressed a subset of these PBRM1 missense mutations in renal cancer cell lines and assessed how they affect PBRM1 binding to acetyllysine substrates, and subsequent renal cancer growth.
Role of locus coeruleus expressing angiotensin type 1 receptors (AT1R) neurons in fear learning and stress‐induced anxiety
Authors: Yu Zhe , Marvar Paul 
Published: May 2022

BackgroundThe renin‐angiotensin system (RAS) has been implicated in stress‐related disorders, however the central mechanisms responsible for this remains unknown. The locus coeruleus (LC), a major noradrenergic nucleus of the brain, plays a critical role in modulating anxiety‐like behaviors. The LC has also been previously shown to express angiotensinogen (AGT), the pre‐cursor for angiotensin, as well as strong expression of angiotensin II receptors, but its role in stress‐related disorders has not been examined. Using angiotensin II type 1 receptor (AT1R)‐eGFP and Cre mice combined with neuroanatomical and behavioral approaches, we examined the role of LC expressing AT1R in fear‐ and anxiety‐related behavior.MethodsRNAscope® in situ hybridization assay was used to analyze cellular mRNA expression of AGT in the LC while in vivo chemogenetics combined with behavioral testing in AT1R‐Cre mice mouse was used to examine the role of LC‐AT1R cells in fear memory and stress‐induced anxiety. Further, dual immunohistochemistry plus retrograde tract tracing was used to characterize LC AT1R‐eGFP+ cells and limbic circuit connections.ResultsAGT mRNA and AT1R‐eGFP+ immunoactivity was found localized to the LC. The majority of AT1R‐eGFP+neurons (94%) in LC were co‐localized with tyrosine hydroxylase, a marker for norepinephrine‐containing neurons. Retrograde labeling revealed that some of these AT1R‐eGFP+ neurons send projections to the basolateral amygdala (BLA), an important brain structure for modulating stress induced fear related anxiety responses. Next, the AT1R+ neurons in LC were silenced using Cre‐dependent inhibitory designer receptor exclusively activated by designer drug (hM4Di DREADD) expression followed by Clozapine‐n‐oxide (CNO) administration. Silencing the LC AT1R‐expressing neurons prior to fear extinction training impaired the extinction of learned fear as shown by increased percent freezing during the training (time × drug interaction, F (7, 70) = 3.219, p<0.01, n=6) (Fig 1). Furthermore, restraint stress‐induced anxiety behavior was attenuated by LC AT1R+ neuron inhibition, as shown by increased center entries (22.7 ± 7.3 Saline v.s. 50.5 ± 8.8 CNO, p<0.05, n=6) and % time in center (2.97 ± 1.14 Saline v.s. 8.6 ± 1.4 CNO, p<0.05, n=6) in the open field test, and increased open arm entries (5 ± 1.3 Saline v.s. 12 ± 1.1 CNO, p<0.05, n=6) in the elevated plus maze test (Fig. 2).ConclusionThese findings provide evidence for a novel angiotensinergic LC cell type and position the LC AT1R as a potential mediator of noradrenergic regulation in learned fear and stress‐induced anxiety. Future studies are needed to fully characterize the underlying neurocircuits and neuropeptide modulators that likely interact with the LC AT1R expressing neurons during stress‐related behaviors.