bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of Innate Immunity in Immune Enhancing Effects of Candida Skin Test Reagent
Authors: Nakagawa Mayumi , Coleman Hannah , Sikes Jim , Daniels Jaclyn , Nagarajan Uma 
Published: June 2021

OBJECTIVESSerial intra‐lesional injections of common warts with skin testing reagents such as Candida are effective in regressing them. The goal of this study was to test our hypothesis that the Candida skin testing reagent enhances anti‐HPV T‐cell responses through activation of pattern recognition receptors (PRRs) on Langerhans cells and subsequent cytokine(s) secretion.METHODSLangerhans cells were treated with Candin® (Allermed, San Diego, CA). LPS and/or zymosan were used as positive controls. Cells were harvested at 8 and 24 hours for RNA extraction, and qRT‐PCR analysis was performed for IL‐1beta, IL‐6, IL‐8, IL‐10, IL‐12p40, IL‐23Ap19, IL‐27p28, IFN‐gamma, and TNF‐alpha. The threshold cycles were normalized to GAPDH expression. In order to remove a small amount of organic solvent (0.4% phenol), Candin was dialyzed using PBS in some experiments. In addition, expressions of PRRs known to associate with Candida albicans (DC‐SIGN, Dectin‐1, Dectin‐2, Galectin‐3, Mincle, mannose receptor, TLRs 1, 2, 4, 6, and 9) were examined. The role of Dectin‐1 was examined by antibody blocking. Supernatants were analyzed for the presence of IL‐12p40 and IL‐12p70 proteins.RESULTSExpression of IL‐12p40 mRNA (≥ 5 fold increase over untreated at 8 hour) was detected most frequently with Candin (15 of 16 experiments using Candin and 5 of 6 experiments using dialyzed Candin). All other cytokines examined were detected in multiple experiments in the following order of decreasing frequency: IFN‐gamma, IL23Ap19, IL‐1beta, IL‐27p28, IL‐6, IL‐8, TNFalpha, and IL‐10. IL‐12p40 and IL‐12p70 proteins were detected. All PRRs examined were detected. Dectin‐1 appeared to have a role in IL‐12 production in some subjects.CONCLUSIONSCandin appears to promote T‐cell responses through inducing IL‐12 production by Langerhans cells, and may be an attractive new adjuvant for HPV therapeutic vaccines.
Interleukin‐33 drives a proinflammatory endothelial activation that selectively targets non‐quiescent cells
Authors: Haraldsen Guttorm 
Published: June 2021

Interleukin‐33 is an alarmin released from stressed or damaged cells. We investigated the effects of interleukin (IL)‐33 on human umbilical vein endothelial cells comparing it to IL‐1β and observing highly similar phosphorylation of signaling molecules and transcription profiles. On the other hand, intradermally injected IL‐33 elicited significantly less endothelial activation when compared to IL‐1β and led us to observe that quiescent endothelial cells (ppRblowp27high) were strikingly resistant to IL‐33. Accordingly, the IL‐33 receptor was preferentially expressed in non‐quiescent cells, corresponding to selective induction of adhesion molecules and chemokines. Multiparameter phosphoflow cytometry confirmed that signaling driven by IL‐33 was stronger in non‐quiescent cells. Manipulation of nuclear IL‐33 expression by siren or adenoviral transduction revealed no functional link between nuclear, endogenous IL‐33 and exogenous IL‐33‐responsiveness.We conclude that in contrast to other inflammatory cytokines IL‐33 selectively targets non‐quiescent endothelial cells. By this novel concept quiescent cells may remain non‐responsive to a proinflammatory stimulus that concomitantly triggers a powerful response in cells that have been released from contact inhibition.
Ethanol‐evoked reduction in myocardial contractility and autonomic dysregulation in freely moving female rats
Authors: Ibrahim Badr Mostafa , Fan Ming , Abdel‐Rahman Abdel A 
Published: June 2021

Acute ethanol reduces blood pressure (BP) in proestrus female rats via reduction in cardiac output, which reflected reduced cardiac contractility based on indirectly measured dP/dtmax (maximal rise of ventricular pressure over time). In this study we directly measured cardiac contractility indices [left ventricular developed pressure (LVDP), dP/dtmax and the ventricular isovolumic relaxation time constant (Tau)] in conscious freely moving female rats via fluid filled catheter. We validated this technique using isoproterenol, which produced the expected dose‐related increases in dP/dtmax. To test whether ethanol cardiac depressant effect precedes the hypotensive response in freely moving female rats, we directly measured cardiac function along with BP, HR, in two animal groups received either intragastric ethanol (1 g/kg) or water (control). Further, we conducted offline analysis of sympathovagal activity (spectral analysis) to elucidate ethanol effect on autonomic balance. Acute ethanol, compared to water, caused slowly developing reduction in BP that was significant at 40 min, reached its nadir at 70 min, and lasted till the experiment ended at 90 min. However, significant reductions in myocardial contractility indices (LVDP and dP/dtmax) started 10 min after ethanol, which preceded the hypotensive response. The high‐frequency (HFα) component of spectral analysis of RRI (cardiac vagal control) increased by intragastric ethanol starting at 20 min and continued throughout the experiment. However, the spectral measure of sympathetic activity, low‐frequency (LFα) component of RRI, was not affected by ethanol resulting in net decreases in the sympathovagal, LFRRI/HFRRI, ratio. Collectively, our findings directly implicate the reduction in cardiac contractility and enhancement of cardiac vagal control, at least partly, in acute ethanol‐evoked hypotensive effect in proestrus female rats.Supported by NIAAA Grant 2R01 AA14441–06
A novel fluorescent ligand binding assay for free fatty acid receptor FFA1
Authors: Watson Sarah‐Jane , Brown Alastair , Holliday Nick 
Published: June 2021

Obtaining affinity estimates at FFA1 receptors using radioligand binding is made difficult by lipophilic ligands. We characterised the properties of a new fluorescent ligand 40Ag‐Cy5 (CellAura) using 293TR cells expressing FLAG‐FFA1. Whole cell competition binding experiments using 40AG‐Cy5 were performed on 96‐well plates, followed by imaging and granularity analysis on a confocal platereader. Curve fitting used GraphPadPrism.Both 40Ag‐Cy5 and its non‐fluroescent precursor 40Ag‐cg were agonists at FFA1 (Fluo4 measurements of intracellular Ca2+), with respective pEC50 values of 5.8±0.2 and 7.9±0.3. In binding experiments 40Ag‐Cy5 (100 nM) was displaced by synthetic ligands such as GW1100 (pIC50 6.0 ± 0.2) and GW9508 (pIC50 6.8 ± 0.2). However oleic acid (up to 100 μM) did not displace 40Ag‐Cy5 binding. By solving the Cheng‐Prusoff correction for GW1100 competition curves using 100nM or 1μM 40Ag‐Cy5 (GW1100 pIC50 5.6), we obtained pKi estimates for 40‐Ag Cy5 (6.2), and GW1100 (6.0; equivalent to its functional pKb). Fluorescence correlation spectroscopy experiments also confirmed 40Ag‐Cy5 binding to FFA1. Thus 40Ag‐Cy5 provides a novel fluorescent ligand which enables estimation of synthetic ligand affinities at FFA1.Supported by EPSRC and AstraZeneca.
Ischemic stroke produces long‐term fiber track myelin loss that contributes to persistent sensory, motor and cognitive behavioral deficits
Authors: Poon Carrie , Zhou Jin , Zhuang Jian , Li Jie , Bloom Jonathan , Ooi Evelyn , Lax Daniel , Rosenbaum Daniel , Barone Frank 
Published: June 2021

ObjectiveTo investigate and characterize long‐term post‐ischemic stroke hemispheric and cellular loss, and fiber track injury. Also to investigate the sensory, motor and cognitive performance deficits for use in future restoration research.MethodsMiddle cerebral artery occlusion (stroke) or sham surgery was performed in male Sprague‐Dawley rats. Neurobehavioral (sensory, motor, beam balance, reflexes, hindlimb and forelimb function, and cylinder placement tests) and cognitive (complex active place avoidance; APA and simple passive avoidance retention; PA tests) performance was measured. Electroretinograms (ERG), ischemic hemisphere, hippocampus neuronal loss and fiber track injury were measured.ResultsStroke surgery produced significant (p<0.01) sustained (10 weeks) deficits in motor, sensory, beam balance and hindlimb performance. Stroke produced large and prolonged cognitive performance deficits in APA learning (p<0.01) but smaller effects on PA retention. No effects on ERG or hippocampus neurons occurred but fiber tracts in the external capsule and striatum exhibited significant (p<0.01) damage.ConclusionSpecific, long‐term deficits in neurobehavioral tests were verified as endpoints for stroke restorative therapeutic research. Fiber track injury contributes to these deficits and is especially important for the complex cognitive control required for APA learning.
Predicting Anthracycline‐induced Cardiotoxicity in Children – Genome‐Wide Association Study
Authors: Aminkeng Folefac , Ross Colin JD , Rassekh Rod S , Brunham Liam R , Weissman Cory , Dube Marie‐Pierre , Visscher Henk , Rieder Michael J , Carleton Bruce C , Hayden Michael R 
Published: June 2021

Identified genetic markers for anthracycline‐induced cardiotoxicity (ACT) explain only a small fraction of the variability of this phenotype, suggesting the presence of other, as‐of‐yet unidentified susceptibility loci. Therefore, our goal is to identify additional genetic markers with large effect size via genome‐wide association study (GWAS).Patients were recruited and clinically characterized (age at start of treatment, cumulative dose, gender, anthracycline and tumor type, radiation therapy involving the heart, follow‐up time and assessment of LV dysfunction) via the Canadian Pharmacogenomics Network for Drug Safety. We have recruited and clinically characterized over 400 patients from across Canada to serve as our discovery cohort and over 120 patients from the Emma Children's Hospital in Amsterdam, the Netherlands, to be used as our replication cohort. More patients are currently being recruited and the clinical characterization is ongoing. We are now genotyping samples with an Illumina GWAS panel and the statistical analyses will be underway soon using SVS/Helix Tree, SPSS, Epi Info, PLINK and R.Novel genetic predictors for ACT may become essential for screening patients before the start of treatment, comprehensive risk assessment and management, and evidence‐based treatment decisions, monitoring and prevention.This work was supported by CIHR, CFRI, and Genome BC.
Microcirculatory anatomy can explain the illusory cooperativity seen in the flow response to exogenous adenosine
Authors: Bindschadler Michael David , Bassingthwaighte James 
Published: June 2021

OBJECTIVETo demonstrate that illusory cooperativity can arise in the microcirculation as a result of competition and spatial localization.Adenosine in the myocardial interstitium binds to adenosine A2A receptors on vascular smooth muscle, leading to reduced tone and increased myocardial blood flow. The relationship between the concentration of supplied adenosine and the increase in flow is very steep (Hill coefficient 5–7), suggesting strong receptor binding site cooperativity. In fact, this apparent cooperativity may be illusory, and can arise solely from the anatomic geometry and cellular composition of myocardial tissue. Along the path from capillary to arteriolar smooth muscle cell, adenosine faces a permeability barrier (the endothelial clefts) and a diffusion distance (back from capillary to arteriole). Cellular transporters and metabolic enzymes compete for adenosine all along this path, influencing not only the apparent ED50 of arterially supplied adenosine, but also the shape of the dose‐response curve. Using a mathematical model, we demonstrate the independent influences of competition and access barriers (permeability barriers and diffusion distances), as well as how these effects can combine to make the dose‐response curve for a single binding site receptor (Hill coefficient 1) appear extremely cooperative.
Anti‐aggregatory effects of Sulforaphane on in vitro platelet function
Authors: Gillespie Scarlett , O'Malley Aislinn , Solomon Antonia , Emerson Michael , Gavins Felicity 
Published: June 2021

IntroductionSulforaphane (SFN) is a phytochemical with proven anti‐inflammatory effects in various inflammatory models. Platelet‐leukocyte‐endothelial cell interactions are integral to the inflammatory cascade. Research has investigated SFN's effects on leukocytes and endothelial cells, but its effects on platelets are previously unknown.AimElucidate the effect of SFN on platelets in‐vitroMethods and ResultsPlatelet aggregation was studied using light transmission aggregometry. Washed platelets (WP) were incubated with SFN (1, 40, 60 and 100μM) for 2/30mins, then challenged with collagen (2.5μg/ml). Incubation with SFN, 2 min (100μM) and 30min (40, 60 and 100μM), prior to collagen‐stimulated decreased platelet aggregation. Similar trends were seen following SFN incubations with platelet rich plasma (PRP) aggregations to ADP (1.56μM), however, this was not significant. Ca2+ mediated platelet signalling showed no detectable Ca2+ mobilisation following SFN addition. Lactate dehydrogenase (LDH) release following SFN incubation showed negligible differences to that associated with vehicle treatment or agonist stimulation.ConclusionSFN inhibited platelet aggregation to collagen in WP, and showed similar trends in PRP following ADP stimulated aggregations. SFN incubation did not appear to be associated with calcium mobilisation events, indicating these anti‐aggregatory effects were independent of intracellular Ca2+ movement, and low level LDH release suggested these effects were not via cytotoxicity. This novel work indicates an anti‐aggregatory ability of SFN in an agonist specific manner.Work funded by the Medical Research Centre, UK.
Experimental Diabetes Impairs Endothelial IKCa Channel Function in Uteroplacental Arteries from Pregnant Rats
Authors: Gokina Natalia I. , Phillips Julie K. , Bonev Adrian D. , Gokin Alexander P. , Goloman Gabriela 
Published: June 2021

Diabetic pregnancy is associated with adverse fetal complications in part due to impaired uteroplacental blood flow. In this study, we explored the role of endothelial cell (EC) SKCa and IKCa channels in impaired uteroplacental vasodilation during diabetic pregnancy. Diabetes was induced by i.p. injection of streptozotocin on 2nd day of pregnancy. We tested the effects of NS309 and CyPPA on arterial diameters, SMC [Ca2+]i and membrane potential using fura 2‐based fluorescence and glass microelectrodes. Whole‐cell patch clamp methodology was used to characterize SKCa and IKCa channel function in isolated ECs of control and diabetic rats. NS309‐induced vasodilation, SMC [Ca2+]i reduction and hyperpolarization were significantly impaired by diabetic pregnancy. Maximal responses to CyPPA were 50–60% smaller than those to NS309, and were not affected by diabetes. NS309 markedly potentiated endothelial K+ currents in ECs from control and diabetic rats. K+ currents were inhibited by appr. 80% and 20% with charybdotoxin (CTX) and apamin, respectively. Only CTX‐sensitive currents were reduced in diabetic ECs (6.5 ± 1.6 vs. 11.1 ± 1.1 pA/pF at 0 mV; n = 7).In summary, IKCa channels play a predominant role in the control of uteroplacental dilatation. Impaired function of IKCa channels contributes to diabetes‐induced endothelial dysfunction in the maternal uteroplacental circulation.Supported by NIH HL088245
EDHFs contribute to ACh‐mediated vasodilation in human skin in a dose‐dependent manner
Authors: Brunt Vienna E , Fujii Naoto , Minson Christopher T 
Published: June 2021

The human cutaneous microcirculation has been shown to be reflective of globalized microvascular function, but the role of EDHFs in acetylcholine (ACh) mediated vasodilation has yet to be explored. In humans and animals, EDHFs appear to play only a modest role in ACh vasodilation, with a greater contribution at higher doses of ACh and under disease or NO‐deficient conditions. Five bolus doses of ACh (0.01, 0.1, 1, 10, and 100mM) were delivered via microdialysis fibers continuously infused with 1) Lactated Ringer's, 2) tetraethylammonium (TEA; calcium‐sensitive potassium channel & EDHF inhibitor), 3) L‐NNA + Ketorolac (to inhibit NO + COX), and 4) TEA + L‐NNA + Keto. The hyperemic response was characterized as area under the curve (AUC) and reported as % max cutaneous vascular conductance. In our preliminary findings (n=5), TEA had no effect on AUC compared to control at any of the lower doses of ACh, but did attenuate AUC to 100mM ACh (Control: 31,621±5,947% vs. TEA: 20,809±6,686%). We found similar results with combined NO + COX inhibition. TEA + L‐NNA + Keto did not attenuate AUC compared to L‐NNA + Keto until the 100mM dose of ACh (L‐NNA + Keto: 16,308±2831% vs. TEA + L‐NNA + Keto: 8,491±1,853%). Therefore, EDHFs appear to contribute to ACh vasodilation in the skin in a dose‐dependent manner, similarly to other vascular beds, confirming the skin to be a good model of overall microvascular function.Supported by NIH Grant HL081671
Post‐translational cleavage of Flt1 occurs very close to transmembrane domain releasing extracellular N‐terminal fragment which binds to PlGF
Authors: Raikwar Nandita S , Liu Kang Z , Thomas Christie P 
Published: June 2021

Vascular endothelial growth factor receptor‐1/fms‐related tyrosine kinase 1 (VEGFR1/ Flt1) is a membrane bound tyrosine kinase receptor that can bind VEGF and PlGF and is involved in angiogenesis and vasculogenesis. Protein kinase C agonist phorbol myristic acid (PMA) stimulates Flt1 mRNA and protein expression in vascular endothelial cells. We studied the regulation of post translational cleavage of tagged full length Flt1 expressed in HEK 293 cells. Flt1 expression and cleavage of its extracellular (EC) domain is robustly induced by PMA and heparin treatment further enhances release of the EC domain into conditioned media. Co‐expression of the metalloproteases ADAM10 or ADAM17 significantly enhanced EC domain cleavage and reduced the abundance of uncleaved Flt1. In contrast, knock down of ADAM10 or ADAM17 reduced Flt1 EC domain cleavage. Conditioned media from Flt1 adenoviral transduced cells bound PlGF reducing free PlGF measured by ELISA. By sequential deletion analysis we identified a 5 AA region adjacent to the Flt1 transmembrane (TM) domain that is necessary to abolish the Flt1 EC domain cleavage. Thus, we demonstrate that proteolytic cleavage of Flt1 occurs close to the TM domain and is regulated by metalloproteases, with release of an EC fragment that is functionally equivalent to sFlt1.NIDDK
Role of peripheral inflammation in central cytokine signaling, depression, and fear
Authors: Tronson Natalie Celia , Radulovic Jelena 
Published: June 2021

Activation of the innate immune system by chronic illness or injury is correlated with depression, anxiety and PTSD. In patients of heart attack (myocardial infarction, MI), rates of major depression and PTSD diagnoses are two‐ and three‐fold higher, respectively, than the population at large. Consistent with other patients, post‐MI incidence of PTSD and depression are correlated with increased levels of circulating pro‐ inflammatory cytokines. How circulating cytokines affect the brain remains unknown.We used a surgical model of myocardial infarction in mice to investigate the role of sustained peripheral inflammation on alterations in (1) fear conditioning and depression‐like behaviors (2) cytokine signaling in the brain and (3) sex differences in post‐ MI behavioral and inflammatory changes.Male, but not female mice exhibited enhanced fear conditioning soon after MI. Both males and females showed impaired memory and depression‐like behavior at later time points. 舲Within the hippocampus and prefrontal cortex, cytokine signaling and JAK/STAT3 were upregulated after MI in both female and male mice. The role of the cytokines and their downstream signaling in behavioral and emotional alterations are currently under investigation.These findings suggest cytokine‐dependent signaling as a candidate mechanism for susceptibility to, or development of depression and PTSD.
Hypothermia through TRPV1 activation provides neuroprotection in acute cerebral ischemic stroke when initiated 90 minutes after reperfusion
Authors: Cao Zhijuan , Balasubramanian Adithya , Marrelli Sean 
Published: June 2021

New treatment options are needed for ischemic stroke that are compatible with the conscious subject. We examined the therapeutic potential of producing hypothermia through activation of the transient receptor potential V1 (TRPV1) channel in a mouse model of focal stroke.Core temperature, BP, and HR were measured in conscious TRPV1 WT and KO mice following subcutaneous dihydrocapsaicin (DHC; TRPV1 agonist) treatment. Subsequently, mild hypothermia (33 °C, >; 8 hours) by DHC infusion was initiated in conscious focal cerebral ischemia/reperfusion (I/R) mice (by distal MCA plus CCA occlusion) at 90 minutes reperfusion. Behavior testing and brain TTC staining was performed after 24 hours reperfusion.DHC produced mild hypothermia in control and I/R mouse models, but not in TRPV1 KO mice. DHC treatment (1.25mg/kg) showed a small transient decrease (6 mmHg) in BP which could be completely blocked by atropine; there was no effect of DHC on HR. DHC‐induced hypothermia initiated at 90 minutes post‐reperfusion significantly decreased the infarct volume and improved behavioral neurofunction.Our data indicate that activation of TRPV1 is a novel and safe method to induce sustainable and reversible hypothermia which provides neuroprotection in acute cerebral I/R mice. We propose that this method may provide a more broadly applicable means to produce therapeutic hypothermia for stroke. Funded by R21NS077413.
Subjective Tinnitus: Morphological Abnormal Features in the Inner Ear
Authors: Tanioka Hisaya , Tanioka Sayaka 
Published: June 2021

BackgroundAs many as 40 million people in the United State have tinnitus. The reported prevalence range is from 7% to 32%. The evaluation of a patient with tinnitus requires a defaulted history and comprehensive audiologic evaluation to determine the appropriate radiologic study of the choice.PurposeDescribe the key anatomic or physiological issues, imaging findings or imaging technique the proposed paper will address. It is important to classify tinnitus as pulsatile or nonpulsatile for treatments. So, we must know how morphological features in the inner ear of subjective tinnitus.Subjects and MethodsImaging studies of 30 patients with clinically suspected nonvascular tinnitus were analyzed from our data. Pt not only arises from the vascular causes but can also originate from nonvascular structures and is classified as nonvascular tinnitus.3D membranous labyrinth technique was obtained by HR‐CT with direct volume rendering by the use of current workstation soft.ResultsUnilateral tinnitus showed the lateral lesion of the affected utricle was swelling. But, the size and shape of maculae were within normal limit within early phase by virtual histological study. And, some cases revealed abnormal image of the cochlear duct such as irregular surface or discontinuity.ConclusionAll tinnitus showed swelling of the affected utricle. This cause may be lack of uniformity of endolymphatic pressure or unbalance of metabolic process of the tissue of swelling wall of the utricle may be occurred. We thought this morphological finding was due to probably both the increased endolympatic pressure and partially decreased blood flow. Indeed, improved cases of subjective tinnitus showed improvement imaging after medical treatments and cardiovascular exercise.TreatmentsIt is conceivable that increased blood flow reduce the increased endolymphatic pressure in the inner ear by achieving a balance of endolymph. Pts take some vascular medicines and cardiovascular exercise.