bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sex bias in drug‐induced immune mediated hepatitis
Authors: Li Zhaoxia , Talor Monica Vladut , Rose Noel R. , Njoku Dolores B. 
Published: June 2021

Many autoimmune diseases are more prevalent in women than in men. Drug hapten‐induced immune‐mediated hepatitis (DIH) from halogenated volatile anesthetics or other drugs is also more prevalent in women than in men. However, precise mechanisms responsible for sex bias are unclear. We investigated sex bias in female and male BALB/c mice by immunizing with hepatic S100 proteins covalently linked to the trifluoroacetyl chloride (TFA) hapten formed from CYP2E1 metabolism of anesthetics. After three weeks we measured hepatitis, serum TFA and S100 antibodies, and assessed liver and spleen supernatant cytokines. We also measured splenic foxp3+CD4+CD25+ regulatory T cells two weeks following immunizations. We found that female mice developed more severe hepatitis and had higher anti‐TFA and −S100 antibodies than males (p < 0.01). Elevated pro‐inflammatory cytokines IL‐2, IL‐9, IL‐12p70, and IL‐17 were found in female livers (p < 0.01). Interestingly, male mice had higher levels of splenic IL‐10 (p < 0.05) and elevated splenic foxp3+CD25+CD4+ T cells at two weeks (p < 0.05). Sex bias in DIH is partially due to elevated anti‐inflammatory cytokine IL‐10 and induced regulatory T cells in males. Further investigations in these areas may uncover therapeutic agents to treat or prevent DIH.Supported by AARDA, Mr. and Mrs. Joseph Scoby, The Gail I Zuckerman Foundation and NIHR21DK075828.
Scheduling process at DEA ‐ the example of cannabidiol
Authors: Ghozland Sandy G , Tolliver James M , Carr Susan M , Sannerud Christine A 
Published: June 2021

DEA collects and reviews scientific, medical and other data on substances to determine their abuse potential and placement into the Controlled Substances Act (CSA). The scientific data reviewed includes in vitro pharmacology and pre‐clinical abuse liability studies. In particular, data on discriminative stimulus effects and rewarding properties is essential to determining whether a substance belongs in a class of drugs already scheduled, and to predict its abuse potential. The medical data reviewed includes laboratory‐controlled clinical studies. Databases and surveys on drug abuse patterns, illicit trafficking and seizures provide DEA with valuable information on the actual abuse of a drug and its consequences on health, public health and public safety.Cannabidiol (CBD) is one of many cannabinoids present in marijuana, and as such is in schedule I of the CSA. DEA is currently conducting a scientific review of CBD to elucidate its pharmacology and abuse liability and to identify gaps in the published literature.
Early environmental exposure to methanol as a sensitization factor for Parkinson's disease: a closer look into the etiology of PD
Authors: Mackey Veronica Renee , Charlton Clivel G. 
Published: June 2021

Parkinson's disease (PD) is a neurodegenerative disorder characterized by a progressive loss of substantia nigra (SN) dopaminergic neurons that project to the striatum. The etiology of PD is unknown; there is the possibility of early exposure to toxicants creating a vulnerability stage for PD to occur later in life. Here, we investigate whether methanol (MeOH) exposure in early life causes sensitization of the basal ganglia to challenges, such as 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP) that precipitates the symptoms of PD. C57/bl6 mice were exposed to sub‐chronic levels of MeOH in utero. At 12 wks of age, the offspring were treated with MPTP for seven days. Tyrosine hydroxylase (TH) protein was analyzed in the midbrain and striatum. MPTP increases the expression of TH in the midbrain and reduces it in the striatum, suggesting that the increase of TH in the midbrain may be due to impairment of its transport from cell bodies in the SN to the terminals in the striatum. Lastly, the fetal MeOH sensitized the mice to MPTP reducing effects, suggesting that fetal exposure to MeOH may be a pre‐disposing factor for PD.This project was supported, in part by NIH Grant 1R21 NS 049623‐01A1
Acute drug‐induced inactivation of the h5‐HT7 receptor: a pharmacological profile
Authors: Knight Jessica , Smith Carol , Toohey Nicole , Teitler Milt 
Published: June 2021

The h5‐HT7R is one of 13 5‐HT receptors expressed in humans. It is involved in the control of cognition, emotion, and cardiovascular tone. Recently we reported that a 30 min exposure to nM levels of the antipsychotic drug risperidone results in an inactivation of the h5‐HT7R‐mediated stimulation of cAMP production (Smith et al, Mol. Pharmacol., 2006). To further investigate this unusual effect of this highly prescribed drug we have been testing other drugs for similar inactivating properties. 9‐OH‐risperidone, the active metabolite of risperidone, and methiothepin, a potent 5‐HT receptor antagonist, also potently inactivate the receptor. Preliminary studies indicate amoxapine, ritanserin, metergoline, lisuride, bromocryptine, SB‐269970 also produce this effect: clozapine, TFMPP and tenilapine did not produce the inactivating effects. It appears that a majority of h5‐HT7 receptor antagonists produce this unusual and dramatic effect. h5‐HT7‐mediated MAPK phosphorylation is also inactivated by risperidone, 9‐OH‐risperidone, and methiothepin, consistent with data indicating an irreversible or pseudo‐irreversible interaction is occurring between the drugs and the receptor. Further studies are underway to investigate the mechanism and in vivo relevance of these observations.Supported by MH56650 (M.T.)
Nitric Oxide Scavenging by Environmental Particles and Engineered Nanoparticles: Size and Composition Dependency
Authors: Nurkiewicz Timothy R. , Dreher Kevin L. 
Published: June 2021

Inhaled particulate matter (PM) elicits systemic biological responses such as impaired arteriolar dilation and increased venular leukocyte adhesion. One mechanism that may contribute to such effects is NO scavenging by pulmonary translocated particles. The purpose of this study was to determine if PM scavenges NO in vitro, and also determine if PM size or composition influences any such activity. PM was classified as combustion source particles, engineered nanoparticles or nuisance dusts. NO solutions were generated by SNAP and were measured with a NO sensor. A concentration of 1–2.5 μg/ml was used for each particle, and responses were normalized to a bolus SNAP dose, [NO]=841 nm. Among fly ashes, residual oil fly ash was the most potent scavenger, decreasing the signal by 67%. Coal fly ash from Western Kentucky and Montana, as well as Mt. St. Helens ash did not significantly alter NO levels. Diesel exhaust particles attenuated the NO signal by 19%. Among nanoparticles, TiO2 decreased the NO signal by 39%. Single and multi‐walled carbon nanotubes did not produce an effect. These findings suggest that air particulate pollution as well as some engineered nanoparticles have the potential to scavenge NO, and this ability is heavily dependent upon particle size, composition and/or surface properties.Support: HEI #4730 and NIH RO1 ES015022 to TRN(This abstract does not necessarily reflect EPA policy)
PACE therapy and its influence on microcirculatory hemodynamics and leukocyte‐endothelial interactions
Authors: Krokowicz Lukasz , Klimczak Alesandra , Duggan William , Grykien Christopher , Mielniczuk Mariusz , Siemionow Maria 
Published: June 2021

Purpose: Effect of Pulsed Acoustic Cellular Therapy (PACE) therapy on muscle microcirculatory hemodynamics, neovascularization, leukocyte‐endothelial interactions and muscle flap healing was assessed.Methods: Cremaster muscles were dissected in 42 Lewis rats, divided into 5 groups: 1) controls (n=10); 2) PACE with 200 or 3) 500 impulses (n=8 each) immediately before dissection; 4) PACE with 200 or 5) 500 impulses (n=8 each) 24h before dissection, followed by microcirculatory hemodynamic recording at 1,2,3,4 h. Immunohistochemistry assessed expression of cell adhesion molecules and proangiogenic factors: VEGF and vWF.Results: PACE increased diameter of first (A‐1 in group 3) and second order arterioles (A‐2 in groups 2 and 3) (P<0.05). In groups 2, 3 and 4 rolling and sticking leukocytes were significantly decreased compared to controls (P<0.05). 200 impulses of PACE increased number of transmigrating leukocytes compared to 500 PACE impulses and to controls (P<0.05). Leukocyte‐endothelial interaction correlated with upregulation of ELAM‐1, CD15s and VCAM‐1. Synergistic expression of proangiogenic factors was significantly upregulated 24h after 500 impulses of PACE.Conclusions: PACE therapy increased main arterioles diameters and has direct effect on activation of leukocyte‐endothelial interactions and stimulation of expression of proangiogenic factors at microcirculatory levels
Fatty acid dependent regulation of renin transcription by nuclear hormone receptor HNF‐4
Authors: Brinkmeier Thomas , Steege Andreas , Fähling Michael , Kirschner Karin , Martinka Peter , Persson Pontus B. , Scholz Holger , Mrowka Ralf 
Published: June 2021

Production and release of renin are pivotal in controlling the Renin‐angiotensin‐system (RAAS), a main long‐term regulator of blood pressure and electrolyte homeostasis. In this context, the regulation of renin transcription by hormonal and nutritional agents is of fundamental interest.Fatty acid dependent transcription factors LXRalpha and PPARgamma have already been shown to affect renin gene transcription. We focussed our attention on another member of the nuclear hormone receptor family, HNF‐4, which is bound by fatty acyl CoA modulating its transcriptional activity. In co‐transfection experiments, HNF‐4 activated a luciferase reporter construct containing 5797bp upstream the transcriptional start site of the renin gene more than five‐fold. This effect could be completely abolished by the addition of omega3 fatty acids at a concentration of 500μM. Using computational methods, we identified five potential binding sites for HNF‐4 in this sequence. There is first evidence that HNF‐4 binds to these sites depending on the presence of fatty acyl CoAs.As HNF‐4 is expressed abundantly in proximal tubular cells our results could shed light on the regulation of local proximal tubular RAAS by fatty acids, indicating a new potential role of HNF‐4 in the kidney. Down‐regulation of renin gene expression may contribute to the antihypertensive effect of fish oil omega3 fatty acids.
Renin release is differentially sensitive to clinically used calcineurin inhibitors
Authors: Friis Ulla G , Madsen Kirsten , Skøtt Ole , Hansen Pernille B , Jensen Boye L 
Published: June 2021

Renin release from juxtaglomerular cells (JG) is paradoxically inhibited by a rise in [Ca2+]i. We hypothesized that the calmodulin‐dependent phosphatase calcineurin in necessary for the inhibitory effect of [Ca2+]i. The hypothesis was tested in isolated JG‐cells by whole cell patch clamp and by renin release studies. In rat JG‐cells, the calcineurin inhibitor cyclosporine A (CsA, 5 μM) increased membrane capacitance (Cm), an index of cell surface area, in single JG‐cells by 10.7% ± 2.9% (n=5; P<0.05). A structurally distinct calcineurin inhibitor tacrolimus did not change Cm significantly when applied directly to JG‐cells. Similarly, renin release from JG‐cell cultures was stimulated by cyclosporine but not tacrolimus. Binding proteins for both inhibitors were detected in afferent arterioles (cyclophilin and FKBP). The effect on Cm was mimicked by the calmodulin‐inhibitor W‐13 (100 μM) in the patch pipette. When the experiments were repeated with a protein kinase A‐blocker (RpcAMPs (5 μM)) in the patch pipette, CsA and W‐13 still increased Cm significantly (15.8 % ± 4.2 % (n = 4) and 9.3 ± 3% (n = 4), respectively) indicating that the increase in Cm is independent on the cAMP‐PKA pathway. We conclude that calcineurin exerts a tonic suppressor effect on renin release from juxtaglomerular cells and that renin release is differentially sensitive to clinically used calcineurin inhibitors.This study was supproted by grants from Carlsbergfondet, the Novo Nordisk Foundation and the Danish Heart Foundation.
Vasomotor changes in controlled and uncontrolled hypertension
Authors: Frances Maria F , Zamir Mair , Usselman Charlotte W. , Goswami Ruma , Masse Nick , Fleischhauer Arlene , Nolan Robert , Shoemaker J. Kevin 
Published: June 2021

The relationship between arterial stiffness and cardiovascular disease has been implicated in the development of isolated systolic hypertension. This study tested the hypothesis that hypertension is related to altered vascular mechanics and hence altered hemodynamics in skeletal muscle. The ability to detect such changes was tested using a lumped Windkessel model that incorporates several parameters of vascular control including resistance (R), compliance (C), viscoelasticity (K) and inertance (L). Two groups of middle aged hypertensive individuals were tested, one with normal blood pressure (NBP) following treatment (n=10), and the other with uncontrolled BP (UBP) (n=27). Baseline beat‐by‐beat measures of heart rate (ECG), mean arterial BP (Finometer), and forearm blood flow (FBF; Doppler ultrasound) were collected. Systolic BP, pulse pressure and mean arterial pressure were higher in the UBP group (p<0.01). Furthermore, values of C (0.008 for NBP vs. 0.006 mL/mmHg for UBP; p<0.02) and K (0.161 for NBP vs. 0.233 mmHg/mL/min for the UBP group; p=0.1) indicated that indeed the vessel walls in the UBP group were less compliant and offered more viscous resistance to stretch. Values of R were the same in the two groups. These findings suggest that hypertensive individuals with controlled BP are better able to maintain the function of skeletal muscle vasculature for muscle perfusion.Supported by the Heart & Stroke Foundation of Canada and the Canadian Stroke Network.
Atrial natriuretic peptide inhibits angiotensin II‐induced cell proliferation in fetal sheep cardiomyocytes in vitro
Authors: O'Tierney Perrie Faye , Chattergoon Natasha , Louey Samantha , Giraud George , Thornburg Kent 
Published: June 2021

Objectives: The anti‐hypertrophic properties of atrial natriuretic peptide (ANP) in cardiomyocytes have been well‐demonstrated; ANP inhibits the effects of several potent hypertrophic agents (ie. Angiotensin II, endothelin‐1, phenylephrine). During the last 1/3 of gestation the fetal heart grows by both proliferation and hypertrophy. We hypothesized that ANP also has an anti‐proliferative effect on cardiomyocyte (CMC) growth during fetal development.Methods: Actively dividing cardiomyocytes were isolated from right and left ventricles of late‐gestation fetal sheep. Cultured fetal CMCs were treated with 10uM Bromodeoxyuridine (BrdU) and increasing doses of ANP (3.25, 32.5, 100, 325, 650 nM) with and without 100nM angiotensin II. Following 48 hours of treatment, cells were fixed and stained for myosin and BrdU. BrdU incorporation was used as an index of cell proliferation.Results: Angiotensin II induced BrdU uptake in fetal CMCs and this was inhibited by ANP in a dose‐dependent manner (P=0.024). Physiological doses of ANP alone have no significant effect on BrdU uptake, though at supraphysiological levels (650nM), ANP increased BrdU uptake by fetal CMCs (P=0.051). No significant differences were seen between ventricles.Conclusion: Physiological doses of ANP inhibit fetal cardiomyocyte proliferation induced by angiotensin II.This research was supported by a NICHD (P01HD34430) and a N.L. Tartar Trust research fellowship.
Inositol 1,4,5‐trisphosphate Pathway Mediates Testosterone‐Induced Cardiac Hypertrophy
Authors: Estrada Manuel , Oyarce Cesar , Toyos Marcela , Altamirano Francisco 
Published: June 2021

The anabolic hormone testosterone is associated with cardiac hypertrophy, but the signal transduction mechanisms involved are poorly known. In this work, primary cultures of neonatal cardiomyocytes where used to study testosterone effects on intracellular Ca2+ and their relation with cardiac hypertrophy markers: β‐MHC and skeletal α‐actin (SKA). Testosterone (100 nM) induced a rapid increase in intracellular Ca2+ that was blocked by inositol 1,4,5‐trisphosphate (IP3) inhibitors (U73122, 2APB and Xestospongin C) or by overexpression of the Ca2+ buffer protein parvalbumin. Neither nifedipine nor ryanodine affected the testosterone‐induced Ca2+ response. Stimulation with testosterone resulted in an increase of β‐MHC (12h) and SKA (24h) expression. Both in the presence of inhibitors of the IP3 pathway or in cells transfected with parvalbumin, β‐MHC and SKA expression were inhibited. Moreover, testosterone induced an increase in NFAT activation measured as luciferase activity that was inhibited by FK506. NFAT inhibition in turn blocked the hypertrophy markers increase. Cyproterone, an inhibitor of androgen receptor, had no effects on the intracellular Ca2+ or NFAT increase, but attenuated β‐MHC and SKA expression. These results suggest a synergism between pathways involving intracellular signal cascades and the classical genomic actions of androgens for testosterone‐induced cardiac hypertrophy.Fondecyt 1060077 FONDAP 15010006
Direct measurement of periarterial nitric oxide (NO) in resistance vessels of aged WKY rats reveals elevated basal and suppressed flow‐mediated production which is normalized by apocynin.
Authors: Zhou Xiaosun S , Bohlen H Glenn , Unthank Joseph L , Miller Steven J 
Published: June 2021

Abnormal regulation of the nitric oxide (NO) system may be responsible for the age‐related impairment in tissue perfusion and collateral growth. Direct, real time, in vivo measurements of NO in aging resistance vessels are not available. This study tested the hypothesis that aging is correlated with reduced basal and flow‐mediated NO production, which can be reversed by anti‐oxidant therapy. Our mesenteric model was used in young (Y) and retired breeder (RB) WKY and RB pre‐treated with apocynin (RB+Apo). NO‐ specific electrodes measured mesenteric periarterial NO simultaneously with arterial flow. After NO baseline measurement, collateral flow was varied from 50–200% of control by placement of microvascular clamps on adjacent arteries. RB had significantly increased baseline NO compared with Y (2053 ± 504 vs. 1059 ± 160 nM, p< 0.05). Periarterial NO levels were altered in proportion to blood flow changes in Y but not in RB rats (Figure 1). In RB+Apo, NO baseline was reduced (760 ± 102 nM) and flow‐mediated NO production restored in RB+Apo (Figure 1). Immunoblotting showed that RB had significantly increased phospho‐eNOS levels compared to either Y or RB+Apo. This suggests that the elevated baseline and suppressed flow‐mediated NO production may result from near maximal activation of eNOS in the RB. eNOS stimulation by reactive oxygen species produced by NADPH oxidase is a mechanism to be explored.Support: HL42898 and HL20605
TIMP3 regulates p38 MAPK‐dependent myogenesis via TACE
Authors: Li Yi‐Ping , Chen Shuen‐Ei , Liu Huijie , Jin Bingwen 
Published: June 2021

Activation of p38 MAPK is a critical step in the onset of myogenesis. However, the upstream signaling mechanism that activates p38 during myogenesis remains undefined. We have recently found that p38 activation in differentiating myoblasts and regenerating muscle is mediated by autocrine TNF‐α release which is the function of TNF‐α converting enzyme (TACE). In the present study, we tested a hypothesis that tissue inhibitor of metallopreoteinase3 (TIMP3), the endogenous inhibitor of TACE, is a physiological suppressor of myogenesis, and is a part of the regulatory mechanism for the activation of myogenesis. We observed a dramatic down‐regulation in TIMP3 level during the first 48 h of C2C12 myoblast differentiation. Overexpression of TIMP3 suppressed TNF‐α release and blocked p38 activation, myogenin and myosin heavy chain expression, and myotube formation. Conversely, addition of recombinant TNF‐α (50 pg/ml) to differentiation medium rescued the impaired myogenesis. Down‐regulation of TIMP3 was also observed in cardiotoxin‐injured mouse soleus muscle; and overexpressed TIMP3 inhibited p38 activation and myogenesis. In addition, the down‐regulation of TIMP3 is accompanied by a decline in TIMP3 mRNA level. These results indicate that TIMP3 is a physiological suppressor of myogenesis and its down‐regulation activates p38‐dependent myogenesis.Supported by National Institute of Health Grant AR049022
Use of a Planar Patch Electrode to Measure NBCe1‐A Function in Mammalian Cells
Authors: Kao Liyo , Abuladze Natalia , Newman Debra , Liu Weixin , Pushkin Alexander , Kurtz Ira 
Published: June 2021

Standard whole cell‐patch clamping is labor‐intensive whereas automated planar patch clamp devices offer the possibility of higher throughput. Planar patch clamp electrodes are also attractive because their geometry facilitates obtaining electrical and optical recordings simultaneously. Previous studies have utilized the Port‐a‐Patch system (Nanion Technologies GmbH), to study ion channel activity however there are no studies which have demonstrated the utility of this approach with electrogenic cotransporters, which in general have a lower conductance. In this study, we utilized a modified Port‐a‐Patch system to measure the functional activity of the electrogenic sodium bicarbonate cotransporter, NBCe1‐A. HEK‐293T cells permanently expressing NBCe1‐A were generated. Cell monolayers were detached using 0.025% Trypsin/0.22 mM EDTA in PBS and were re‐suspended at a concentration of 500,000 cells/ml in 50% DMEM/PBS. The cells were kept at room temperature until measurements were made. Approximately 2,500 cells were placed on a planar electrode chip from which a single cell was automatically selected. Planar electrode chips with variable resistances were assessed. Using chips with a resistance of 2 – 3.5 MOhm, we routinely obtained GOhm seals using this approach. Whole cell currents were elicited by voltage ramps from −100 mV to +30 mV from a holding potential of −100 mV. Cell membrane capacitance for the HEK‐293T cells was 7.14 ± 0.67 pF (n = 21) and access resistance was 8.6 ± 0.5 MOhm (n =21). The DIDS‐inhibitable Na+‐and HCO3−‐dependent NBCe1‐A‐mediated average whole cell current was 29.3 ± 3.2 pA (n =21) at +30 mV. These results represent the first reported use of a planar patch‐clamp device to study ionic currents mediated by a heterologously expressed electrogenic cotransporter in mammalian cells.Supported by the NIH
Graduate courses in statistical and model‐based analysis of physiological data
Authors: Moore Leon C , Solomon Irene C , Clausen Chris 
Published: June 2021

We have developed two modular graduate courses in statistical and model‐based analysis of physiological data. The motivation was to provide students with a common set of skills and tools that facilitate their research training. Each course lasts 5 weeks and has 2 hours of lecture and 2 hours of lab per week. The first course, Statistical Analysis of Physiological Data, consists of a review of statistical methods used in laboratory research. It focuses on the interaction between experimental design and statistical analysis. To pass, the students must analyze and present a set of 15 problems. The lab sessions consist of tutorials on software usage, reference database searching, graphic design and data presentation, as well as student presentations of the assigned problems. The second course, Model‐based Analysis of Physiological Data, focuses first on nonlinear curve fitting and how it is used to analyze transporter kinetics and band density on gels. The course concludes with sessions on programming in which the students are required to write a program that numerically solves a 4‐compartment model of lithium pharmacokinetics. Again, the students are given a set of problems that they must complete and present to pass the class. The courses have been well received by the students, and have markedly increased their ability to analyze data and solve quantitative problems.Funded by NIH Grant DK066124