bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of a Humanized Anti‐Cocaine Monoclonal Antibody on the Pharmacokinetics of Cocaine in Rats and Mice
Authors: Gooden Felicia CT , Tabet Michael R , Bartley Erin L , Ball William J , Norman Andrew B 
Published: June 2021

Monoclonal antibody (mAb) h2E2 is a further humanized version of the high‐affinity anti‐cocaine mAb 2E2. h2E2 is a potential immunotherapeutic for cocaine abuse that retains the same high‐affinity (4 nM Kd) and specificity as 2E2 for cocaine over its inactive metabolites. Infused h2E2 (1.6 μmol/kg i.v.) in both mice and rats produced dramatic increases in the area under the plasma cocaine concentration‐time curve (AUC) and a concomitant decrease of >;90% in the brain AUC following an i.v. injection of an equimolar cocaine dose. Consequently, h2E2 substantially reduced cocaine's volume of distribution in both species by sequestering cocaine in plasma, indicating potential clinical efficacy. Furthermore, h2E2 did not inhibit the clearance of cocaine in either rats or mice. Interestingly, h2E2 restricted cocaine distribution and metabolism from plasma to a single‐compartment model in mice but cocaine disappearance remained multi‐compartmental in rats. This differential effect of h2E2 on cocaine distribution between species may be related to the metabolism of cocaine predominantly to ecgonine methylester (EME) in mice but to benzoylecgonine (BE) in rats. The metabolism of cocaine to BE and EME in humans is between that in mice and rats. Thus, these studies will help determine whether differences in cocaine metabolism are relevant to h2E2's clinical efficacy.Supported by NIH grant DP1DA031386.
Effects of some angiotensin‐converting enzyme inhibitors and angiotensin II receptor blockers on experimental inflammations
Authors: Magyar Ioan , Cret Mihai , Cuparencu Barbu 
Published: June 2021

MethodThe experiments were carried out in Wistar albino rats, weighing 150–200 g. The inflammations were induced as folows: 1. the hind paw edema evoked by the intraplantar injection of 0.1 mL from a 10% aqueos kaolin suspension; the measurements of the paw volume increases were performed plethismometrically; 2. the insertion of a cotton pellet, weighing 50 mg into the scapulo‐humeral region. The following bood inflammatory markers were measured: fibrinogen and C‐reactive protein (CRP). The tested drugs by intraperitoneally route were: captopril (0.4–4.0–6.0 mg), enalapril (0.2–2.0–4.0 mg), lisinopril (0.2–2.0–4.0 mg), quinapril (0.2–2.0–4.0 mg) and valsartan (4.0–8.0–16.0 mg). The statistical evaluation was carried out by chi square and “size effect” procedures.ResultsAll ACEI exhibited in a dose‐dependent and a time‐related manner a proinflammatory activity. The most active compounds were: quinapril and enalapril and lisinopril. Valsartan was ineffective. Fibrinogen and CRP blood levels were increased by all ACEI. However, no clear dose‐response relationships might be seen.DiscussionConclusionFrom these data it may be inferred that the proinflammatory activity of ACEI was due to an accumulation of bradykinin, a phlogogenic peptide. It is well known that the converting enzyme, besides its splitting effect of angiotensin I, leading to the genesis of angiotensin II has a kininase activity.
FORMULATION AND OPTIMIZATION OF SOLID DISPERSION AS A SUSTAINED RELEASE MATRIX FOR ORAL KETOROLAC TROMETHAMINE
Authors: Abdul Rasool Bazigha K. , Aziz Uday S. , Abu‐Gharbieh Eman , Khan Saeed 
Published: June 2021

ObjectiveIn the present study, solid dispersion is employed in the formulation of sustained‐release matrix particles of KT with chitosan (CT) as a carrier.MethodsThe matrix solid dispersion was prepared in three different ratios of KT: CH (1:3, 1:5, and 1:7) by spray‐drying method. The matrix particles were characterized by angle of repose, compressibility, scanning electron microscopic (SEM), FTIR and differential scanning calorimetry. The release profile from the matrix particles was studied at pH 1.2 and 6.8. In vivo biological screening in mice and rats for the analgesics, anti‐inflammatory and ulcerogenicity was also performed.ResultsThe formation of composite particles was confirmed by SEM. DSC and FTIR spectroscopy analysis suggested an interaction between the carbonyl group of KT and the amino group of CH formed a hydrogen bond. Optimum sustained in vitro release, retained analgesic and anti‐inflammatory activities with significantly reduced ulcerogenicity was obtained from the matrix particles (1:7) compared to the parent drug.ConclusionKT‐CH solid dispersion prepared by spray‐drying is useful as a sustained release preparation.
Matrix Metalloproteinase 2 (MMP‐2) Increases Atherosclerotic Aorta Rings Vasoconstriction.
Authors: Azevedo Aline , do Prado Alejandro Ferraz , Pernomian Laena , Bendhack Lusiane Maria , Gerlach Raquel Fernanda 
Published: June 2021

The present study aimed to evaluate the vascular function of atherosclerotic aortic rings after intraluminal injection of MMP‐2. Thoracic aorta isolated from New Zealand rabbits with atherosclerosis (feed 1% cholesterol for 8 weeks) and controls (normal diet) were isolated and tested for vascular reactivity function after intraluminal exposure to Krebs solution (vehicle) or MMP‐2 at 1.2 μg/mL, at 37º C for 30 minutes. Curves for Acetylcholine (ACh) and Phenylephrine (PE) were performed in endothelium‐intact rings. The vascular contraction was normalized by grams of tension by dry tissue (g/g) and the relaxation by % of PE contraction. In the vascular reactivity of atherosclerotic vessels, MMP‐2 increased Emax of PE (1842,08±138,77g/g, n=6) versus vehicle (1432,28±124,13g/g, n=7; p<0.05), but no differences were observed on Emax between vehicle in control vessels (1416,13±120,87g/g, n=7) and MMP‐2 in those vessels (1550,44±110,93g/g, n=7). Atherosclerotic vessels filled with vehicle had an impaired relaxation to ACh (80,98±4,63%, n=7) compared to control vessels filled with vehicle (99,86±3,13%, n=7), and MMP‐2 had no effect on relaxation. Since MMP‐2 was able to increase the effect of PE, it can be speculated that it may worsen the endothelial dysfunction of atherosclerotic arteries.Supported by the State of Sao Paulo Research Foundation (FAPESP), SP, Brazil.
Conditioning of human embryonic stem cell‐derived endothelial cells with PBMCs confers TLR4 sensing in co‐culture conditions
Authors: Reed Daniel Mark , Foldes Gabor , Gashaw Hime H , Harding Sian E , Mitchell Jane A 
Published: June 2021

Human embryonic stem cell‐derived endothelial cells (hESC‐EC) are being investigated as potential therapies for vascular disease. We have previously shown that hESC‐EC do not have functional Toll‐like receptor‐4 (TLR4) responses but do respond avidly to cytokines such as TNFα and IL‐1β1,2, which illustrates that these cells do have fully functional TLR signaling via MyD88 and NF‐κB. In this study we show that, whilst hESC‐EC do not respond to LPS (1μg/ml) to release CXCL8 when treated in mono‐culture, they mediate a synergy when co‐cultured with peripheral blood mononuclear cells (PBMCs). No synergy was found in co‐cultures stimulated with TNFα (10ng/ml) suggesting that the conditioning effects of PBMCs on hESC‐EC is above the level of NF‐κB and downstream pathways leading to CXCL8 release. This data is highly novel but preliminary and more studies need to be performed to establish how this conditioning effect is mediated. These findings will be significant in the use of hESC‐EC in vessel engineering and highlight the importance of cell interactions on stem cell and endothelial cell function.This work was funded by the Welcome Trust, BHF and an MRC studentship awarded to DMR.
Heart rate responses to osteopathic manipulations in healthy young men
Authors: Ioudina Marina , Toder Eric , Boudreaux Brittany 
Published: June 2021

Osteopathic manipulative treatment (OMT) is used to treat cardiac patients. Some manipulations are hypothesized to affect the cardiovascular system by altering autonomic tone, although physiological effects of individual techniques have not been well defined. Previous studies from our laboratory have shown that cranial manipulations (CMs) as well as articulatory (A) and inhibitory (I) rib raising techniques decreased cardiac output in healthy individuals. This study measured responses to individual manipulations on heart rate (HR) in healthy men in order to further our understanding of OMT effects on cardiovascular physiology.25–35‐yr old men participated. HR was measured in response to manipulations thought to stimulate parasympathetic activity (CM), stimulate (A), and inhibit (I) sympathetic activity. ECG recordings were used to measure changes in HR. Manipulations were performed by an osteopathic physician.CMs caused a significant 6% decrease in HR during manipulations. Both (A) and (I) techniques did not have a significant effect on HR.These data demonstrate that CMs have a significant effect on HR, likely via an increase in parasympathetic activity. No effects of (A) and (I) rib raising manipulations on HR responses were observed. Future studies should include a larger number of subjects of various age groups.
Effects of training status on circulating angiogenic cell paracrine activity in young men and women
Authors: Landers‐Ramos Rian Q , Jenkins Nathan T , Spangenburg Espen E , Hagberg James M 
Published: June 2021

PURPOSETo determine if a relationship exists between physical fitness and the ability of CD34+ or CD34−/CD31+ circulating angiogenic cells (CAC) to influence angiogenic function through paracrine activity.METHODSPeripheral blood was drawn from healthy inactive (n = 7), active (n = 10) and endurance‐trained (n = 7) men and women aged 25–39. Mononuclear cells were isolated by density centrifugation and CD34+ and CD34−/CD31+ fractions were purified by immuno‐magnetic selection. Conditioned media (CM) from cultured CACs was retrieved and used for a HUVEC‐based angiogenesis tube formation assay. Tube length and complexity were imaged and quantified.RESULTSCM from CD34+ CACs of endurance‐trained subjects resulted in 21% greater tube length (P<0.05) and 48% greater complexity (P<0.05) than inactive subjects, and 22% greater complexity than active subjects (P<0.05). CD34+ CM from active subjects produced greater length and complexity than inactive subjects (P<0.05, 18% and 33%, respectively). Tube length was 22% greater and complexity was 42% greater (P<0.05 each) with CD34−/CD31+ CAC CM of endurance‐trained subjects compared to inactive individuals.CONCLUSIONSThese preliminary data provide promising support for a chronic exercise‐induced increase in paracrine‐mediated angiogenic function of CD34+ and CD34−/CD31+ CACs.Support: NIH AGT3200068 and UMD KNES Graduate Research Initiative Fund
Inhibition of the mammalian Target of Rapamycin (mTOR) does not alter nitric oxide or superoxide levels in human skin
Authors: Zhao Joan L , Roman Linda J , Wu Yubo , Johnson John M , Kellogg Dean L 
Published: June 2021

In laboratory animals, mTOR inhibition for 7 days causes significant reductions in nitric oxide (NO) levels with concomitant increases in superoxide production. We hypothesized mTOR inhibition by rapamycin would effect similar changes in humans. 8% rapamycin ointment (RAPA) was applied twice daily for 7 days to an 8×8 cm area of skin on the abdomen while a similar second site was treated with the ointment vehicle only. 5 middleaged persons participated. Selective amperometric electrodes placed intradermally within both skin areas monitored NO levels. Superoxide levels were monitored by perfusing the intradermal microdialysis probes at both skin areas with a solution of saline containing oxidized ferricytochrome c and analyzing for reduced cytochrome c in the collected perfusates. Superoxide in the cutaneous interstitial space diffuses across the probe membrane, forming reduced ferrocytochrome c and altering the spectrophotometric absorbance.ResultsNO levels at RAPA treated sites averaged 811±72nM and 526±56nM at vehicle treated sites. These values did not differ between RAPA and vehicle treated sites (p>;0.50 between site). Superoxide levels at RAPA sites averaged 11.73±2.17μM and 11.29±1.90μM at vehicle treated sites. These levels did not differ between sites (p>;0.40). We conclude that chronic RAPA treatment does not alter NO or superoxide levels in human skin. (supported by NIH Grant AG041365)
Muscle‐specific VEGF knockout disrupts thermoregulation without altering mitochondrial morphometry or activity
Authors: Mandler William Kyle , Audet Gerald N , Olfert I Mark 
Published: June 2021

VEGF is believed to enhance thermogenesis in response to cold exposure through promoting angiogenesis, enhancing metabolism, and upregulating mitochondrial biogenesis. Using muscle‐specific VEGF gene deleted mice, we tested the hypothesis that loss of VEGF in skeletal muscle will affect thermoregulation.VEGF knockout (KO) and wild‐type (WT) mice were exposed to 4°C for 6 hours. Core temperature was measured in 15 minute increments. Flow cytometry analysis was performed on isolated subsarcolemmal (SSM) and interfibrillar (IFM) mitochondrial subpopulations taken from cardiac and triceps surae muscle tissue of KO or WT to determine mitochondrial size and complexity.Initial temperature was lower in the KO. During cold exposure, temperatures dropped more quickly in the KO group. The average time to reach a core temperature decrease of 8°C was shorter in the KO group. There were no differences in skeletal muscle SSM or IFM in size or complexity. In the cardiac tissue, both SSM and IFM exhibited greater size in the KO group. There were no significant differences in mitochondrial density for any tissue or subpopulation.Muscle‐specific VEGF KO mice are unable to maintain core temperature to the same degree as WT in the face of a 4°C challenge and have a lower core temperature at room temperature. Skeletal muscle mitochondrial density and morphology does not appear to play a role in this effect.
Molecular mechanisms underlying aflatoxin‐induced mutagenesis
Authors: Lin Ying‐Chih , Li Liang , Burgers Peter M. , Stone Michael P. , Lloyd R. Stephen 
Published: June 2021

Abstract Aflatoxin B1 (AFB1) is a known mutagen and carcinogen that is associated with hepatocellular carcinoma (HCC). Although the underlying mechanisms contributing to these events are far from complete, our investigations provide insight into the molecular mechanisms of aflatoxin‐accelerated carcinogenesis. Site‐specific mutagenesis assays in primate cells were used to examine the mutagenic potential of the primary and persistent AFB1‐DNA adducts, AFB1‐N7‐Gua and AFB1‐FAPY‐Gua, respectively. Both AFB1‐N7‐Gua and AFB1‐FAPY were highly mutagenic at frequencies of 45% and 97%, respectively, with the predominant base substitution being G to T transversions, a mutation that is consistent with observations made in aflatoxin‐associated HCC samples. Moreover, we present the first biochemical evidence that the replicative polymerase δ and the specialized translesion synthesis (TLS) polymerase ξ4 could bypass AFB1‐N7‐Gua accurately, while the other two TLS polymerases κ and ξ were responsible for G to T mutations that were induced by AFB1‐N7‐Gua and AFB1‐FAPY.This work was supported by NIH R01 CA055678.
Submaximal cycling exercise stimulates mTOR signaling pathway in human skeletal muscle
Authors: Kakigi Ryo , Naito Hisashi , Yoshihara Toshinori , Ozaki Hayao , Kobayashi Hiroyuki , Okada Takao 
Published: June 2021

It is known that eccentric muscle contraction promotes muscle protein synthesis. However, effects of concentric contraction on muscle protein synthesis are not clear. We determined whether high repetition of concentric contraction by cycling exercise stimulates mTOR signaling pathway in human skeletal muscle. Six young subjects (21±1 years) performed 90% VO2max cycling exercise at 90rpm for 5 minutes on bicycle ergometer. Muscle biopsies (vastus lateralis) were obtained before exercise and, at 1 and 3 h post‐exercise. mTOR signaling and markers of autophagy (LC3BI and LC3BII protein) were analyzed by using western blotting. Plasma GH, IGF‐1 and insulin levels significantly increased in post‐exercise period (p<0.05). Akt phosphorylation, upstream of mTOR, significantly increased at 1 h post‐exercise (p<0.05). S6K1 phosphorylation, a marker of mTOR activity, significantly increased at 3 h post‐exercise (p<0.05). At the same time point, LC3BII to I ratio were significantly decreased (p<0.05). These data suggest that high repetition of concentric contraction such as cycling exercise can stimulate mTOR signaling in human skeletal muscle, which might leads to promotion of protein synthesis and attenuation of autophagic protein degradation.This study was partly supported by the Sasakawa Scientific Research Grant from the Japan Science Society, and a Grant‐in‐Aid for Young Scientists (B) No. 24700703.
Intramuscular and interstitial nitric oxide concentrations following doxorubicin administration in nude mice.
Authors: Ryan Talia , Fabris Sergio , MacLean David A 
Published: June 2021

The purpose of the present study was to examine intramuscular (IM) nitric oxide (NO) concentrations in the gastrocnemius (G) and quadriceps (Q) muscles as well as interstitial NO levels in the G of immunodeficient nude mice (n=6) following the administration of doxorubicin (10mg/kg). Muscles were collected at day 3 and 7 post injection and interstitial concentrations were determined using the microdialysis technique. Gastrocnemius IM concentrations of NO were increased (P<0.05) from control (626±18 μmol/kg dw) to 823 ±48 and 831±22 μmol/kg dw at day 3 and 7, respectively. Similarly, IM quadriceps concentrations of NO increased (P<0.05) from control (599±13 μmol/kg dw) to 738 ±18 and 756±46 μmol/kg dw at day 3 and 7, respectively. In contrast, G interstitial NO concentrations did not change (P>;0.05) from control values (14.0±2.5 μmol/l) for either collection period (day 3 = 16.4±1.7. day 7 = 15.7 ±1.6 μmol/l). These data clearly demonstrate that intramuscular NO concentrations are increased as a result of doxorubicin administration, and remain elevated for up to 7 days following initial dosing. Interestingly, doxorubicin administration did not alter interstitial NO concentrations suggesting that the effects of anthracyclines on stimulating NO production was isolated to intramuscular mechanisms. Overall, this study suggests that doxorubicin administration stimulates intramuscular NO production and may be a result of increased oxidative stress.Supported by NSERC
Changes in exercise hyperpnea are more attributable to limb movement frequency than pedal loading
Authors: Caterini Jessica Elizabeth , Wells Greg , Duffin James 
Published: June 2021

The significance of limb movement frequency in determining ventilation during exercise remains a contested topic in respiratory research. We used two sinusoidal exercise protocols on a cycle ergometer to separately assess the effects of limb movement frequency i.e. cadence (CP) and pedal load (LP) on ventilation. During the first test, cadence was altered sinusoidally while pedal load remained constant. In the second test, pedal load changed sinusoidally while cadence was constant. Variables were fitted to a sinusoid with parameters amplitude, mean, period, and phase angle lag. Five participants completed both protocols. Mean (SD) cadence amplitudes differed between protocols (CP: 23.5 (0.67) rpm, LP: 0.47 (0.3) rpm, p<0.001). The phase lag angles for ventilation also differed, with sinusoidal cadence significantly smaller than sinusoidal load (CP: 23.7 (12.7) deg., LP: 48.1 (14.6) deg., p= 0.023). However the mean ventilation was not different between tests (CP: 44.5 (3.9) L/min, LP: 46.8 (4.2) L/min, p=0.99). VO2 and VCO2 did not differ between tests; eliminating the possibility that changes in ventilatory response between protocols resulted from differences in metabolism. Therefore we concluded that changes in exercise hyperpnea are more attributable to limb movement frequency than pedal loading.Funding provided by the Faculty of Kinesiology and Physical Education at the University of Toronto
Diindolylmethane Attenuates TGF β Mediated Human Lung Fibroblast Proliferation
Authors: Galam Lakshmi , Bandyopadhyay Sayantani , Martinez Osvaldo , Abuelenen Toaa , Castillo Annie , Parthasarathy Prasanna , Lagishetty Venu , Lockey Richard , Kolliputi Narasaiah Kolliputi 
Published: June 2021

ObjectiveIdiopathic pulmonary fibrosis (IPF) is a progressive scarring disorder characterized by the deposition of extracellular matrix causing impaired gas exchange with excessive proliferation and apoptosis‐resistant state of lung fibroblasts. Therefore, inhibitors of fibroblast proliferation offer considerable therapeutic promise. Diindolylmethane (DIM) is found in cruciferous vegetables and has been known for its anti‐cancer and anti‐proliferative properties. Surprisingly, DIM has never been studied in pulmonary fibrosis. In the current study, we tested the effect of DIM on human lung fibroblast proliferation and apoptosis.MethodsHuman lung fibroblasts were treated with TGF β, a fibroblast proliferation inducer, in the presence or absence of DIM. Proliferation and apoptosis were assessed by using commercially available kits. To further understand the mechanism of DIM, cell lysates were analyzed for gene expression analysis using RT PCR. Protein expressions were determined by western blot analysis.ResultsOur results showed that DIM suppressed TGF β induced proliferation in human lung fibroblasts but not apoptosis. In addition, DIM induced MMP1 expression and suppressed TGF β induced alpha smooth muscle actin and thrombospondin‐2 gene expression. Interestingly, DIM is not influencing ADAM‐TS2 expression (a pro‐fibrotic gene) in human lung fibroblasts.ConclusionsOur findings demonstrated that the natural dietary compound DIM attenuates TGF β mediated pro‐fibrotic effects through regulating of MMP‐1 and thrombospondin‐2 in human lung fibroblasts. These findings suggest that DIM may be a potential therapeutic target for pulmonary fibrosis.
P2X receptor‐dependent erythrocyte damage by α‐hemolysin from Escherichia coli triggers phagocytosis by THP‐1 cells
Authors: Fagerberg Steen K , Skals Marianne , Leipziger Jens , Praetorius Helle A. 
Published: June 2021

The pore‐forming exotoxin α‐hemolysin from E. coli causes a significant volume reduction of human erythrocytes that precedes the ultimate swelling and lysis. This shrinkage results from activation of Ca2+‐sensitive K+ (KCa3.1) and Cl− channels (TMEM16A) and reduced functions of both of these channels potentiate the HlyA‐induced haemolysis. This means that Ca2+‐ dependent activation of KCa3.1 and TMEM16A protects the cells against early haemolysis. Simultaneous to the HlyA‐induced shrinkage the erythrocytes show increased exposure of phosphatidyl‐serine (PS) in the outer plasma membrane leaflet, which is known to be a keen trigger for phagocytosis.We hypothesize that exposure to HlyA elicits removal of the damaged erythrocytes by phagocytic cells. Cultured THP‐1 cells as a model for erythrocytal phagocytosis was verified by a variety of methods including live cell imaging. We consistently found the HlyA to very potently trigger phagocytosis of erythrocytes by THP‐1 cells. The HlyA‐induced phagocytosis was prevented by inhibition of KCa3.1, which is known to reduce PS‐exposure in human erythrocytes exposed to both ionomycin and HlyA. Moreover, we show that the P2X receptor inhibition, which prevents the cell damages caused by HlyA, also reduced that HlyA‐induced PS‐exposure and phagocytosis. Based on these results, we propose that erythrocytes, damaged by HlyA‐insertion, are effectively cleared from the blood stream. This mechanism will potentially reduce the risk of intravascular hemolysis