bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD40 activation is angioproliferative in pulmonary microvascular endothelial cells
Authors: Baker Jeffrey T , Moore Timothy M 
Published: June 2021

IntroductionEndothelial CD40 is important during inflammation as an angiogenic stimulus. We tested whether CD40 expression in PMVEC is associated with angioproliferative potential.MethodsWestern analysis for CD40 expression was performed. Growth curves, barrier measurements, and scratch‐wound assays were carried out using an ECIS apparatus (Applied Biophysics) and a digital image system composed of an inverted microscope.ResultsBasal CD40 expression was high in PMVEC, whereas expression was absent in rat pulmonary arterial‐derived EC (PAEC). CD40 expression could be up‐regulated in PAEC following over‐expression of NAP‐1, an endothelial progenitor cell‐associated protein. Exposure of PMVEC to sCD40L (1 μg/mL) for 1 ‐ 48 hours increased proliferation and improved barrier integrity. Exposure to sCD40L also produced an accelerated PMVEC migration response with decreased wound closure times. These effects were inhibited by pre‐treatment with anti‐CD40 antibodies.ConclusionsOur data indicate activation of PMVEC‐expressed CD40 results in angioproliferative responses. These responses may be inherent to cell phenotypes expressing NAP‐1. Identification of PMVEC expressing CD40 in situ may localize pulmonary resident endothelial progenitor cells. Support: Parker B. Francis Foundation and AHA (0765421B) to T.M.M.
Maternal low protein diet: Developmental origin of adult hypertension
Authors: Goyal Ravi , Leitzke Arthur , Goyal Dipali , Longo Lawrence D. 
Published: June 2021

ObjectiveWe hypothesized that maternal isocaloric, low‐protein, diet during pregnancy programs the Renin Angiotensin System in the fetus leading to hypertension in the adult. We conducted studies on pregnant mice, on isocaloric, normal or low protein diet (50% and 33% less protein).MethodsProteins and mRNA levels were measured using western blot and real time PCR technique and blood pressure was recorded by the non‐invasive tail cuff system.ResultsMaternal low protein diet (MLPD) resulted in low birth weight and rapid catch up growth in both male and female offspring. MLPD induced hypertension in female mice offspring at 11 weeks of age, whereas no hypertension was observed in male offspring by 20th week of age. Our results indicate increased AGT and ACE, but decreased AT2, expression of mRNA, with no change in the protein expression of AGT but decreased protein expression of both, ACE and AT2 receptors with MLPD. Promoter methylation analysis of ACE gene showed hypo‐methylation of the CpG islands as a consequence of maternal protein deprivation.ConclusionsOur studies establish that MLPD programs hypertension in female offspring by epigenetic programming of the renin‐angiotensin system.
Effects of Metabolic Syndrome (MetS) on Male and Female Ossabaw Miniature Pig Detrusor Muscle Contractility and Compliance
Authors: Francis Samuel J , Yang David , Sharma Richa , Alloosh Mouhamad , Sturek Michael S , Packer C Subah 
Published: June 2021

Diabetic bladder dysfunction is not understood. Previous work suggests that male MetS Ossabaw pigs have stiffer and more contractile detrusor muscle compared with lean male pigs. The purpose of the current study was to determine if altered detrusor muscle function in MetS pigs is gender dependent. Detrusor strips from male and female Ossabaw pigs maintained on an atherogenic or control diet were mounted at slack length in muscle baths. Following equilibration, muscle strips were stimulated with supramaximal electrical field stimulation (EFS). Following relaxation, cycles of incremental stretches (0.5‐1.5 mm) followed by EFS‐induced contractions were repeated every 5 minutes until maximum total tension was achieved or the tissue yielded. Normalized passive and active length‐tension (L‐T) curves were generated. Both mean passive and active L‐T curves for MetS bladder muscle are elevated above the respective control curves indicating increased contractility and decreased compliance of bladder muscle in both male and female MetS pigs. Stiffer bladders that are hypercontractile could explain both incomplete voiding and increased urinary frequency experienced in diabetes characteristic of metabolic syndrome.(D Yang is an Indy Project SEED Scholar; S Francis is an IUSM Summer Research in Academic Medicine Scholar; R Sharma was a 2007 APS Undergraduate Summer Research Fellow; Support: NIH RR013223, HL062552)
The large conductance Ca2+‐ and voltage‐activated K+ channel is the primary K+ channel that controls cell excitability and contractility in human urinary bladder smooth muscle
Authors: Hristov Kiril L. , Chen Muyan , Kellett Whitney F. , Rovner Eric S. , Petkov Georgi V. 
Published: June 2021

We examined the large conductance Ca2+‐activated K+ (BK) channel expression, electrophysiological properties, and possible physiological roles in human urinary bladder smooth muscle (UBSM). RT‐PCR with freshly isolated human UBSM cells shows mRNA expression of BK channel pore‐forming α, and regulatory β1/β4 subunits. The expression of all three subunits was confirmed by specific antibodies and immunocytochemistry on single cells. The majority of the whole‐cell outward current induced by voltage‐step depolarizations (‐40 to +80 mV) was inhibited by iberiotoxin (IBTX), a BK channel selective inhibitor. Voltage‐dependent spontaneous transient outward currents (STOCs) were recorded from ‐40 to +40 mV. STOCs were blocked by IBTX or ryanodine. In current‐clamp mode, IBTX caused ~3 mV depolarization. In whole cell configuration, in the absence of STOCs, single channels of 139.5 pS conductance with voltage‐dependent amplitude and open probability were recorded. These channels were blocked by IBTX, consistent with the BK channel. In isolated strips, IBTX increased spontaneous phasic contraction amplitude, force, and tone as well as the amplitude of electrical field induced contractions. In conclusion, the BK channel in the human bladder consists of α, β1 and β4 subunits and is the primary K+ channel that controls cell excitability and contractility of human UBSM.Supported by NIH DK‐070909 to G.V. Petkov.
Cardiac Microenviroment Program Human Aminotic Fluid‐derived Stem Cell To Cardiac Phenotype
Authors: Guan Xuan , Delo D. M. , Wang Z. , Soker Shay 
Published: June 2021

ObjectiveIt is believed that certain microenvironment can induce differentiation of stem cell into a specific cell lineage. Therefore, we carried out in vitro coculture experiments to investigate the potential of human amniotic fluid‐derived stem cell (hAFS) to differentiate along cardiac lineage. Our long‐term goal is to explore the feasibility of using this cell source for cardiac cell therapy.MethodsNeonatal rat Cardiomyocytes (NRVM) were obtained by enzymatic digestion of ventricles from neonatal Sprague‐Dawley rats and plated on 1% gelatin coated coverslips. hAFS cells were labeled by PKH26, cultured directly with NRVM. At different time interval, cells were stained with cardiac specific antibodies. For dye transfer experiment, NRVM were preloaded by calcein‐AM and cocultured with PKH26 labeled hAFS.ResultsAs early as 24 hours after initiation of coculture, contracting PKH26‐positive cells were observed. However, their number was low. After 14 days of coculture, immunostaining detected PKH26 positive cells colocalized with cardiac myosin heavy chain, cardiac Troponin T and connxin 43 positive cells. We further conducted dye transfer experiments to test coupling between NRVM and hAFS cells. 18 hour after initiation of coculture, calcein‐AM and PKH26 dual positive cells was observed, suggesting the existence of coupling between the two cell types, possibly through connexin 43.
Tendinous Architecture of Extensor Digitorum
Authors: Singh Keerti , Kesavi D. , Rajendran Melani 
Published: June 2021

OBJECTIVETo study the tendinous architecture of extensor digitorum during routine cadaveric dissections.METHODThirty (30) human cadavers were observed during routine dissections in the Department Of Anatomy of SRMC & RI, Chennai, India. The tendons of the extensor digitorum of these cadavers were examined on both sides for the number of extra slips. The origins and insertions of the extensor digitorum muscles were also observed.RESULTSOut of 30 cadavers, extra slips were observed in only thirteen (13) cadavers (43.3%). The adjacent tendons were connected by intertendinous slips.CONCLUSIONSThe middle finger or the ring finger received doubled or tripled slips, which is a new finding. The index finger received only a single slip. No extra slip was found to the little finger or the thumb. The intertendinous connections were found to be normal as reported earlier. These accessory slips can be used in procedures of tendon rerouting by the plastic surgeons.Grant Funding SourceUniversity
The ligaments and articular surfaces of the talocalcaneal joint: a cadaveric study
Authors: Billard Kathryn , Padfield Beverly , Hrycyshyn Alan , Johnson Marjorie 
Published: June 2021

IntroductionThe articulation between the talus and the calcaneus, the subtalar joint, is important in chronic ankle instability. The relationship between the ligaments and the articular surfaces of this joint has not been thoroughly discussed in the literature. This study aims to describe and quantify the relationship between the morphology and attachments of the interosseous talocalcaneal ligament (ITCL) and the total surface area and configuration of the joint facets on the calcaneus.Methods20 subtalar joints from embalmed cadavers will be dissected and distracted. Measurements of the ITCL and the joint facets on the calcaneus will be reported.ResultsPreliminary results suggest a positive correlation between thickness of ITCL and total surface area of the talar facets on the calcaneus.DiscussionSubtalar joint instability may contribute to recurrent ankle sprains. Knowledge of the ligaments and articular surfaces of the joint is critical for effective diagnosis and treatment. Reconstruction of the ITCL could be crucial in the treatment of chronic ankle instability. Precise information about the morphology, attachments and relationship of the ITCL to the joint facets is necessary to maximize the positive results of reconstructive surgery. This work was supported by the Anatomy & Cell Biology Department at the University of Western Ontario and the Ontario Graduate Scholarship program.Grant Funding SourceOGS
Comparative histological methods for counting optic nerve fibers of the great horned owl, Bubo virginianus
Authors: Perkins Ashley Renee , Grady Shauna M , Walters Linda M , Suchocki Lawerence , Hall Margaret I 
Published: June 2021

Optic nerve fibers are the axons of retinal ganglion cells, and as such they can provide information about the visual acuity and sensitivity capabilities of an animal. Currently, consistent optic nerve fiber counts are not available for most avians. Those that are available are dependent on method, with different methods yielding different counts. Here, optic nerve fibers are counted at light level for the Great Horned Owl. Juxtaposed sections of a single optic nerve are stained using three different techniques: Bielschowsky's silver impregnation, Bodian's protargol, and Luxol Fast Blue stain. The optic nerve is also observed using H&E to demonstrate details of microanatomy. We test two hypotheses: 1) different staining methods yield different myelinated optic nerve fiber counts due to variations in nerve fiber staining; and 2) hand counts and computer‐generated counts will vary depending on the stain. Counts are estimated for the nerve from 25 digital pictures taken at 1000X at regular intervals. Hand counts are performed multiple times by different observers and compared. Computer‐generated counts are performed in Image J. Research support comes from Midwestern University.Grant Funding SourceAAA Annual Meeting Student/Postdoctoral & Young Faculty Travel Award Student Award
Attaching and Effacing Pathogen Infection Causes an Increase in Functional Connexin 43 Hemichannels to Generate Diarrhea.
Authors: Lin Ann En‐Ju , Li Yuling , Vogl A. Wayne , Bechberger John , Naus Christian , Finlay B. Brett , Guttman Julian A. 
Published: June 2021

The attaching and effacing (A/E) pathogens enterohemorrhagic Escherichia coli and enteropathogenic E. coli cause serious diarrheal diseases. These organisms colonize the apices of the intestinal mucosa and inject pathogenic effectors into host cells altering sub‐cellular epithelial cell components. During these infections small molecules are released from the cells into the lumen of intestinal tracts, contributing to diarrhea. We investigate the role of gap junctions, a group of intercellular channels involved in the transport of small molecules and water, during A/E pathogen infection. Gap junctions are composed of paired connexon channels; however single connexon hemichannels also exist. Each connexon is formed by connexin (Cx) monomers. Using Citrobacter rodentium, a natural murine A/E pathogen, we show that Cx43 protein levels are increased and re‐localized during infection. Cx43 that normally localizes only to the lateral membrane of colonocytes was also localized at the infected cell apices forming functionally open hemichannels. During C. rodentium infection, Cx43 mutant mice had abolished diarrheal phenotypes while maintaining the bacterial colonization levels of wild‐type infections. These results provide the first evidence of connexon hemichannel formation to generate diarrhea and provides a novel target for future therapeutics to prevent bacterially‐induced diarrheal disease.Grant Funding SourceCIHR and NSERC
Ability of n‐acetyl cysteine (NAC) to reverse TGF‐beta driven myofibroblast phenotype
Authors: Lu Daryn L , Vaughan Melville 
Published: June 2021

The purpose of this study was to determine whether n‐acetyl cysteine (NAC) is able to reverse the Transforming Growth Factor‐β (TGF‐β) stimulated alpha‐smooth muscle actin (α‐sma) expression generated by myofibroblasts. Normal Carpal Tunnel fibroblasts and Dupuytren's diseased fibroblasts were grown on coverslips at a concentration of 0.5 x 105 cells per coverslip to allow sufficient time for growth to avoid quiescence. Three cell lines were used to help establish statistical significance along with one control.This experiment was conducted in two phases. The first phase involved a control and a group treated with TGF‐β. All initial cell lines were grown in the presence of TGF‐β for an incubation period of 48 hours to stimulate the growth of myofibroblasts. The cells were then fixed and stained with immunofluorescence for a protein characteristic of α‐sma. After the initial incubation period, cells were harvested to determine how much TGF‐β increased α‐sma to establish a base line for future comparison. The second phase involved the group treated with TGF‐β to be split and treated with two additional treatments. One group continued with additional TGF‐β. The other group was treated with both TGF‐β and NAC to determine whether the base line increased, decreased or remained constant after an incubation time of 48 hours. After all cell groups were fixed and stained, the data was quantified and analyzed for statistical significance using an Analysis of Variance test (ANOVA).It was expected that the group treated with additional TGF‐β would produce numerous quantities of myofibroblasts. It was also expected that the second group treated with both TGF‐β and NAC would remain constant without a significant increase in the myofibroblast population. Early results indicated NAC can reverse TGF‐β effects.Grant Funding SourceUniversity of Central Oklahoma Office of Research and Grants
Liver α2,6‐sialylation by ST6Gal‐1: Ablation of the hepatic promoter alters the α2,6‐sialylation pattern of serum glycoproteins but not the overall degree of sialylation
Authors: Migliore Marina Theresa , Jones Mark , Lau Joseph , Kazim Latif 
Published: June 2021

The sialyltransferase ST6Gal‐1, which mediates the synthesis of the α2,6‐sialyl linkage to Gal( ?1,4)GlcNAc, is expressed ubiquitously but with particular abundance in the liver. The acute phase response, the liver's response to insult or injury, results in addition of heavily sialylated proteins into circulation. The acute phase response is also accompanied by hepatic up‐regulation of ST6Gal‐1 expression, but the functional relevance of hepatic ST6Gal‐1 remains unclear.Previously, we generated a mutant mouse, Siat1ΔP1, by inactivation of the P1 region in the liver of the ST6Gal‐1 gene. Past experiments have shown myelopoetic disruption involving altered sialylation of serum glycoproteins. Initial examination demonstrated an essential unchanged degree of overall sialylation between wild‐type and δP1 animals, and serum glycoproteins in animals with globally inactivated ST6Gal‐1 remain fully sialylated. We use 2D gel electrophoresis, lectin affinity, and silver staining for analysis of serum glycoproteins to reveal significant qualitative and quantitative changes in the sialylation of serum glycoproteins between genotypes. These changes were further magnified after subcutaneous exposure of the animals to turpentine. We pay special attention to these mouse models at three crucial time points. Thus, the extent and types of sialyl linkages on serum glycoproteins are important.
Hexabromocyclododecane decreases the lytic function and ATP levels of human natural killer cells
Authors: Hinkson Natasha C , Whalen Margaret M 
Published: June 2021

This study investigates the effect of Hexabromocyclododecane (HBCD) on human natural killer (NK) cell lytic function, additionally; it investigates the effects of HBCD on ATP levels in NK cells. Human NK cells are a subset of lymphocytes that are capable of lysing tumor cells, virally infected cells, and antibody‐coated cells. Hexabromocyclododecane (HBCD) is a non‐aromatic, brominated cyclic alkane used primarily as an additive flame retardant. If HBCD interferes with NK cell function, this could increase risk of tumor development and/or viral infection. NK cells were exposed to various concentrations of HBCD for 24 h, 48 h, and 6 days before determining lytic function and ATP levels. ATP levels and lytic function were also determined in NK cells that were exposed to HBCD for 1 h followed by 24 h, 48 h, and 6 days in HBCD‐free media. The results indicated that exposure of NK cells to 10 µM HBCD for 24 h causes a very significant decrease in both NK cell ATP levels and lytic function (90.5% and 93.5%, respectively). Exposure of NK cells to 10 µM HBCD for 1 h followed by 24h in HBCD‐free media also showed a very significant decrease in lytic function (89.4%) and a slight decrease in ATP levels (46.1%). The results indicate that HBCD exposures decrease ATP levels and lytic function in NK cells.Supported by NIH grant SG06 GM008092‐33
Inhibition of fatty acid synthase by peptides from hydrolysates of β‐conglycinin enriched soybean
Authors: Roman Maxine , Martinez‐Villaluenga Cristina , Bringe Neal A , Gonzalez de Mejia Elvira 
Published: June 2021

The synthesis of fatty acids is a target pathway for chemotherapy development because it has been linked to the growth and survival of cancer cells. The inhibition of fatty acid synthase (FAS) causes apoptosis of cancerous cells. Due to the chemical instability and negative side effects of existing FAS inhibitors, it is important to search for alternative inhibitory sources. The objectives of this study were to study the inhibitory effect of soybean hydrolysates (SH) derived from β‐conglycinin (BC) enriched soybean on FAS and characterize the responsible peptides. Total RNA from 3T3‐L1 adipocytes was isolated and FAS RNA expression was measured using a real‐time quantitative PCR method with SYBR green fluorescence dye. SH (100 μM) derived from soybean enriched BC (up to 47% total protein) showed a marked decrease of FAS mRNA abundance relative to the negative control. However, SH from 24% BC did not affect FAS gene expression (P < 0.0001, R2 = ‐0.90). It was concluded that soybean enriched in β‐conglycinin generates peptides that down regulate gene expression of enzymes involved in fatty acid synthesis in vitro, which can inhibit FAS. The potential health implications of this observation warrant further investigation.Supported by The Monsanto Company, Cargill Co. and University of Illinois
Angiogenesis from endothelial progenitor cells is co‐regulated by vascular endothelial growth factor and matrix stiffness.
Authors: Hanjaya Putra Donny , Truitt Rachel , Yee Jane , Yee Derek , Ceci Doug , Gerecht Sharon 
Published: June 2021

Angiogenesis from endothelial progenitor cells is co‐regulated by vascular endothelial growth factor and matrix stiffness. Endothelial Progenitor Cells (EPCs) in the circulatory system have been suggested to maintain vascular homeostatis and contribute to adult vascular regeneration. These processes require that EPCs break down the extracellular matrix (ECM), migrate, differentiate into endothelial cells, and line the lumen of the blood vessels. Evidently the ECM plays a critical role by providing biochemical and biophysical cues that regulate cellular behavior. Using our controllable in vitro model of angiogenesis we show that vascular endothelial growth factor (VEGF) and matrix stiffness regulate angiogenesis from EPCs. When exposed to low levels of VEGF, EPCs proliferate independently of matrix stiffness. Exposure to high VEGF concentrations results in upregulation of MMPs, demonstrating EPCs migratory and invasive abilities. Under these conditions, the stiffness of the matrix affects the progression of angiogenesis. Immunofluorescence and TEM images confirm the formation of vacuoles, which fully develop into lumen within a chain elongated cell on the softest matrix. RNAi studies further suggest that EPCs sense matrix stiffness through signaling cascades downstream of integrin ligation leading to the activation of the RhoGTPase and Cdc42. These results suggest that dual regulation of VEGF and matrix compliances are required to modulate angiogenesis dynamics of EPCs.Grant Funding SourceAHA
Survey of biological active molecules in the Chinese traditional herbal formula Liu Wei Di Huang Wan for the treatment of Type II Diabetes
Authors: Mao Sena , Chuang G. , Tam W. , Tam G , Tam Chick 
Published: June 2021

ObjectiveIdentify biological active molecules in the ancient herbal formula Liu Wei Di Huang Wan (LWDHW) to treat diabetes in China.MethodsHandbook of Active Constituents from Chinese Materia Medica & Phytomedicine (ISBN: 9620731549) by Wing and Gen.ResultLWDHW (aka Rehmannia Six) is consist of six herbs: Rehmannia glutinosa (Shou Di Huang), Dioscorea opposita (Shan Yao), Poria cocos (Fu Ling), Paeonia moutan (Mu Dan), Cornus officinalis (Shan Zhu Yu) and Alisma plantago (Ze Xie). The emperor herb Rehmannia glutinosa contains jionoside A1 & B1, jiocarotenosides, four iridoid glucosides (catalpol, leonuride, aucubin, and melittoside); and four glycosides (rehmannioside A, B, C, and D) from the roots of species of var. purpurea and hueichingensis; glycosides (desrhamnosyl acteoside, forsythiaside, purpureaside A and B) from the leaves of species of Digitalis purpurea; and glycosides (purpureaside A & B & C, ateoside) from the callus of the same species.LWDHW improves hyperglycemia and increases insulin sensitivity.ConclusionLWDHW has the bioactive function for the treatment of diabetes based on Traditional Chinese Medicine (TCM).