bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of scutellarin on vascular impairment by hypoxia reoxygenation in isolated coronary artery from rat (LB562)
Authors: Yang Weimin , Chen Yajuan , Wang Lei , Li Yang , Chen Chen , Qing Chen 
Published: June 2021

Scutellarin (SCU), a flavonoid from a traditional Chinese medicinal plant, is clinically used to treat cardiovascular diseases. In this study, we investigated the vasoprotective effects of SCU against hypoxia reoxygenation (HR) in isolated rat coronary artery (CA). HR treatment significantly impaired acetylcholine (ACh,0.001‐100 µM)‐induced endothelium‐dependent relaxation in isolated CA rings and these effects were reversed by pre‐incubation with SCU (500 µM). SCU pre‐incubation did not affect ACh response in normal CA without HR treatment. HR treatment did not significantly altered nitroglycerin (NTG, 0.001‐10 µM) induced endothelium‐independent vasodilation in CA and SCU (500 µM) had no significant effect on NTG response in CA with or without HR treatment. The PKG inhibitor Rp‐8‐Br‐cGMPS (4 µM) blocked both the vasorelaxative effects of SCU in CA rings without HR treatment and the vasoprotective effects of SCU in HR‐treated CA. These results demonstrate that SCU repairs HR‐induced vascular impairment in endothelium‐dependent vasodilation and might be via activation of the PKG pathway.Grant Funding Source: Supported by grants from the National Natural Science Foundation of China (Nos. 30960450 and 81173110) and Yunnan Provincial Science and Technology Department (Nos. 2011FA022, 2012BC012, 2008CD054, and 2008ZC111M).
Activation of G‐protein Coupled Estrogen Receptor 1 Protects the Heart Against Ischemia/Reperfusion Injury by Inhibiting the mPTP Opening Via MEK/ERK/GSK‐3β Pathway (LB564)
Authors: Harris Tanoya , Singh Harpreet , Olde Bjorn , Leeb‐Lundberg Fredrik , Toro Ligia , Stefani Enrico , Bopassa Jean Chrisostome 
Published: June 2021

Introduction: Estrogen effect can be mediated by three receptors: Classical estrogen receptors: alpha (ERα) and beta (ERβ), and recently identified G protein‐coupled estrogen receptor1 (GPER1).Hypothesis: We investigated the role of ERα, ERβ and GPER1 in mediating rapid estrogen‐induced cardioprotection in male mice hearts subjected to ischemia/reperfusion using wild type (WT) and gene specific knockout animals.Methods: Isolated hearts from wild type (WT: C57BL/6NCrL), ERα‐/‐, ERβ‐/‐ and GPER1‐/‐ were perfused using Langendorff apparatus with Krebs Henseleit buffer (control) or with the addition of estrogen (40 nM). Hearts were subjected to 18 min global ischemia followed by 60 min reperfusion. Cardiac function was recorded during the entire experiment and myocardial infarct size measured by TTC staining at the end of the reperfusion. Mitochondria calcium retention capacity (CRC) required to induce the mitochondrial permeability transition pore (mPTP) opening was assessed after 10 min reperfusion. Protein levels were measured by Western Blot in whole heart lysates after 5 min treatment just before ischemia, and after 10 min reperfusion. LY294002 and U0126 were used as inhibitor of PI‐3K/Akt, and MAPK/ERK translocation, respectively.Results: In WT, ERα‐/‐ and ERβ‐/‐, estrogen treatment significantly improved cardiac functional recovery, reduced infarct size and improved mitochondrial CRC. However, estrogen effects were completely absent in GPER1‐/‐. Estrogen treatment during 5 min before ischemia induced up‐regulation of Akt, GSK‐3β, and ERK1/2 phosphorylation in WT mouse as compared with control but not in GPER1‐/‐. U126 abolished estrogen effect on mitochondrial CRC while LY294002 could not prevent estrogen effect on GSK‐3β observed in WT. P<0.05 and n=3‐6.Conclusion: Rapid activation of GPER1 induces cardioprotection effect against ischemia/ reperfusion injury. Estrogen effects through GPER1 are associated with phosphorylation of GSK‐3β and ERK1/2, and inhibition of the mPTP opening.Grant Funding Source: American Heart Association, NIH
Active hexose correlated compound moderates lps‐induced gut injury in rats (LB576)
Authors: Doursout Marie‐Francoise , Segal Graeme , Ahn Stephanie , Devakottai Sundar , Liang Yangyan , Wakame Koji , Sundaresan Alamelu , Kulkarni Anil 
Published: June 2021

Introduction: We hypothesize that Active Hexose Correlated Compound (AHCC), a fermented mushroom extract, restores immune function in lipopolysaccharide (LPS) induced inflammation in the gut, especially when endogenous nitric oxide (NO) signaling pathway is impaired. We identified the molecular mechanisms in the gut in rats subjected to LPS‐induced immune dysfunction, treated and non‐treated with AHCC, a potent promoter of immune support.Method: Six rats were instrumented under anesthesia to record mean arterial blood pressure (MAP) and heart rate (HR) and for drug administrations. Following recovery, conscious animals were pretreated with AHCC in drinking water (6%) prior to be subjected to LPS; 20 mg/kg; IV to induce inflammation. MAP and HR were continuously recorded for 3 hrs. following LPS administration. Simultaneously to hemodynamic measurements, blood samples were collected for NO and cytokines production. Animals were sacrificed at 3 hrs. and gut harvested to assess histology.Results and Conclusions: Ourpreliminary data have shown that AHCC induced reductions in LPS‐induced hypotension by 20% and gut edema by 30%. Cytokines e.g. TNF‐α, TGF β and IL‐1 were reduced by 40% in LPS rats pre‐treated with AHCC. Administration of AHCC prior to LPS challenge ameliorates LPS‐induced microvilli disruption in the gut. In summary, our data show that AHCC restored immune function in rats subjected to inflammation and specifically decreased excessive inflammation/oxidative stress in the gut.Grant Funding Source: None
A Humanized Anti‐Cocaine Monoclonal Antibody Decreases the Urinary Excretion of Cocaine from Mice (LB588)
Authors: Gooden Felicia , Tabet Michael , Nieman Michelle , Ball William , Norman Andrew 
Published: June 2021

The recombinant humanized monoclonal antibody (mAb) h2E2 is a lead candidate for immunotherapy of cocaine abuse. This anti‐cocaine mAb has been shown in vivo to inhibit cocaine’s distribution to mouse brain by 19‐fold compared to controls, with an equivalent increase in plasma cocaine concentrations. Despite binding to h2E2 in plasma, cocaine appears to be eliminated more rapidly than in controls. However, in vitro studies using pH‐buffered (1:1) mouse plasma demonstrated that h2E2 protects cocaine from metabolism to the major inactive metabolites of cocaine, ecgonine methyl ester (EME) and benzoylecgonine (BE). As cocaine and its metabolites are also excreted renally, tests for abuse measure urine concentrations of cocaine and BE. It is hypothesized ‐ based upon h2E2’s high affinity and effects on cocaine’s distribution and metabolism ‐ that the presence of h2E2 would decrease urinary excretion of cocaine, EME, and BE. This hypothesis was tested in male Swiss‐Webster mice pre‐treated with an i.v. infusion of vehicle or h2E2 (1.6 µmol/kg) 1 hour prior to i.v. bolus injection of an equimolar dose of cocaine HCl. Excreted urine was collected 17‐20 minutes after cocaine administration and cocaine, EME, and BE concentrations were determined using GC/MS. The mAb h2E2 caused dramatic 8‐fold (88%), 7.7‐fold (87%), and 3.6‐fold (72%) decreases in urinary cocaine, BE, and EME concentrations, respectively, as compared to levels in the absence of h2E2. If similar effects occur in humans, caution should be used when interpreting efficacy measures of anti‐cocaine vaccines and mAbs that are based on urine cocaine and metabolite concentrations.Grant Funding Source: Supported by NIH grant DP1DAP31386.
Role of glucocorticoid receptor and pregnane X receptor in dexamethasone induction of rat hepatic aryl hydrocarbon receptor nuclear translocator and NADPH‐cytochrome P450 oxidoreductase (LB597)
Authors: Riddick David , Hunter Sarah , Vonk Alex 
Published: June 2021

The aryl hydrocarbon receptor (AHR) nuclear translocator (ARNT), as the AHR’s heterodimerization partner, and NADPH‐cytochrome P450 oxidoreductase (POR), as the key electron donor for all microsomal P450s, are important players in the adaptive and toxic responses to polycyclic and halogenated aromatic hydrocarbons. Expression of both ARNT and POR in rat liver is induced by dexamethasone (DEX), a synthetic glucocorticoid known to activate both the glucocorticoid receptor (GR) and the pregnane X receptor (PXR). To study the role of GR in ARNT and POR regulation, we treated male rats with a low DEX dose (0.5 mg/kg), shown to activate GR selectively, and a high DEX dose (50 mg/kg), shown to activate both GR and PXR, in the absence and presence of the GR antagonist RU486 (50 mg/kg). The role of PXR was assessed by comparing the responses to similar low (1 mg/kg) and high (50 mg/kg) DEX doses in wild‐type and PXR‐knockout rats. In both studies, rats were euthanized at 6 and 24 h after DEX dosing. The induction of ARNT mRNA levels by low‐dose DEX and the increase in ARNT protein levels by high‐dose DEX were prevented by RU486, whereas the induced levels of ARNT mRNA and protein did not differ between wild‐type and PXR‐knockout rats. RU486 did not inhibit the induction by DEX of POR expression at the mRNA, protein, and catalytic activity levels. The induction of POR mRNA levels by low‐ and high‐dose DEX was significantly attenuated in PXR‐knockout rats and the increase in POR protein levels and catalytic activity by high‐dose DEX was completely absent in PXR‐knockout rats. These results are consistent with important roles for GR in the DEX induction of rat hepatic ARNT and PXR in the DEX induction of rat hepatic POR.Grant Funding Source: Supported by the Canadian Institutes of Health Research
The Pharmacokinetics/Pharmacodynamics of Resveratrol and Mechanism‐based PK‐PD Modeling Research (LB598)
Authors: Chen Hui , Liu Xinxia , Wan Heng , Zeng Can 
Published: June 2021

Objective: The aim of this research was to develop a mechanism‐based pharmacokinetic‐pharmacodynamic(PK‐PD) model that could quantitatively characterize the time‐dependent relationship between the PK and PD of Resveratrol(Res) in melanoma tumor‐bearing mice for elucidation of its functional mechanisms and molecular targets. Methods: The plasma concentration and AUC of Res and its main metabolites (Res‐glucuronide,Rse‐G; Res‐sulfate,Res‐S) were respectively fitted to the activities, mRNA and protein levels of tyrosinase and COX‐2 for screening of PK input and PD endpoint. Results: The administration of Res could reduce the growth speed of tumor, inhibit the activities of tyrosinase and COX‐2, and down regulate the mRNA and protein expression of COX‐2 in tumor‐bearing mice. Sigmoid Emax model combined with exponential equation was fitted to the PK‐PD modeling for the effects of Res treatment on tumor volume, simple Emax model was fitted to the PK‐PD modeling for the effects on enzyme activity, mRNA and protein expression of COX‐2, while sigmoid Emax model was fitted to the effects on activities of tyrosinase. Conclusion: Res‐G and Res‐S may play roles in the in vivo activities of Res, which could be attributed to their own activities or the deconjugation reactions of metabolite in tumor tissues. PK‐PD models were developed for the effects of Res treatment on enzyme activity of tyrosinase and COX‐2, as well as mRNA and protein levels of COX‐2, which could quantitatively describe the correlation between the PK and PD of Res, Res‐G and Res‐S.Grant Funding Source: Supported by National Natural Science Foundation of China (30973586)
Effects of Diabetes‐Like Altered Glycemia on BBB Gating Functions: Impact on Efflux Transporters (LB600)
Authors: Sajja Ravi , Cucullo Luca 
Published: June 2021

Altered glycemic conditions, such as those observed in diabetic patients, impair blood‐brain barrier (BBB) endothelial physiology and serve as potential risk factor for the onset and progression of major CNS disorders (such as Alzheimer’s disease). In this study, we assessed the impact of diabetes‐like hyper and hypoglycemic conditions on the expression and functionality of major BBB efflux transporters (P‐glycoprotein and MRP4). Confluent monocultures of hCMEC/D3 endothelial cells (a well characterized human BBB endothelial cell line) were exposed to normal (5.5mM), hypo (2.2mM) ‐ or hyper (35mM)‐glycemic conditions of D‐glucose for 3, 6, 12 and 24 h. Following treatment, cells were either lysed or fixed for subsequent protein expression and distribution analysis by western blotting and immunofluorescence. Further, parallel hCMEC/D3 cultures were chronically exposed to identical experimental conditions, but for a period of 3 days, and profiled for genome‐wide transcriptional changes using Illumina® DNA microarray. Our results indicate an up‐regulation of P‐glycoprotein (P‐gp) and MRP4 protein following 12 ‐24 h acute exposure to hypoglycemia (~1.1 fold increase, vs. control), while no differences were observed at 3 h. Hyperglycemic exposure resulted in a time‐dependent alteration of P‐glycoprotein and MRP4 expression with an initial increase at 3 h followed by restoration to normal (MRP4) or down‐regulation (P‐glycoprotein). These results were further confirmed by using L‐glucose as an osmotic control. Gene chip analysis revealed a moderate increase of P‐gp and MRP4 gene expression following repeated exposures to hyperglycemia. In contrast, chronic exposure to cycles of hyper ‐ hypoglycemia significantly decreased P‐gp gene expression (≍ 50% vs. control) while the MRP4 expression was mostly unaffected. In summary, our data demonstrate that acute or recurrent exposures to hypo‐ and hyperglycemia potentially alter the protein/gene expression of major BBB efflux transporters that would impact the BBB integrity and gating function as well as brain drug disposition. Additional studies are currently in progress to determine the changes in the functionality of these transporters in relation to their altered expression.Grant Funding Source: Supported by A.R.D.F and in part by NIH/NIDA R01‐DA029121‐01A1 to LC
Targeted temperature management with novel adenosine agonist (LB604)
Authors: Bailey Isaac , Bogren Lori , Drew Kelly 
Published: June 2021

The decreased metabolism and body temperature (Tb) of hibernating mammals may increase resistance to loss of bone density during prolonged periods of disuse and ischemia/reperfusion injury through targeted temperature management. The A1 adenosine receptor (A1AR) agonist N6‐cyclohexyladenosine (CHA) induces a torpor‐like state in rats, however, CHA has not been tested in humans and full A1AR agonists have cardiovascular risks. Capadenoson (CAP) is a partial A1AR agonist designed to minimize cardiovascular risk and is currently approved for use in humans. The study objective was to test the hypothesis that CAP would decrease Tb in rats in a manner similar to CHA with less cardiovascular risk as a potential means to decrease Tb and metabolism through non‐invasive measures.Procedures were approved by UAF IACUC. Adult, male Sprague‐Dawley rats (n=10), instrumented with IP iButton dataloggers (Maxim Integrated, San Jose, CA) were housed at 23°C on 12L:12D and fed ad lib. Rats were moved to a Ta of 17°C 20‐24h prior to treatments then moved back to Ta of 23°C 24h after treatments with no additional supplemental warming. Tb was monitored at 10min intervals, HR was monitored every 2h using a Littmann© Model 4000 electronic stethoscope (3M, St. Paul, MN) and Beat Counter app (http://vetapps.co.uk). Treatments (vehicles, 1mg/kg CHA; 1 and 2 mg/kg CAP) were administered IP per a balanced block design with 1 week between tests.CHA decreased Tb by 1°C to 17°C, and HR by 12 to 80% from baseline; the time to return to baseline varied. CAP gave more consistent but modest responses with Tb and HR drops of 1.3 and 1.6°C and 24 and 30% for 1 and 2mg/kg doses. All rats returned to baseline levels without evidence of stress indicated by porphyrin accumulation around the eyes.In contrast to expectations, full and partial A1AR agonists differed significantly in effects on Tb and HR in adult rats.Grant Funding Source: Supported by NIH R15NS070779, Alaska Space Grant Program, Undergraduate research fellowship
Estrogen sulfotransferase(est/sult1e1) promotes human adipogenesis (LB606)
Authors: Ihunnah Chibueze , Philips Brian , Ravuri Sudheer , Gibbs Robert , Kirisci Levent , Rubin J , Marra Kacey , Xie Wen 
Published: June 2021

The estrogen sulfotransferase (EST/SULT1E1) is known to catalyze the sulfoconjugation and deactivation of estrogens. The goal of this study is to determine whether and how EST plays a role in human adipogenesis. By using human primary adipose derived stem cells (ASCs) and whole fat tissues from the abdominal fat of obese and non‐obese subjects, we showed that the expression of EST was low in pre‐adipocytes but increased upon differentiation. Overexpression and knockdown of EST in ASCs promoted and inhibited differentiation, respectively. The pro‐adipogenic activity of EST in humans was opposite to the anti‐adipogenic effect of the same enzyme in rodents. Mechanistically, EST promoted adipogenesis by deactivating estrogens. The pro‐adipogenic effect of EST can be recapitulated by using an estrogen receptor (ER) antagonist or ERα knockdown. In contrast, activation of ER in ASCs inhibited adipogenesis by decreasing the recruitment of the adipogenic peroxisome proliferator activated receptor(PPARγ) onto its target gene promoters, whereas ER antagonism increased the recruitment of PPARγ to its target gene promoters. Linear regression analysis revealed a positive correlation between the expression of EST and body mass index (BMI)as well as a negative correlation between the ERα expression and BMI. We conclude that EST is a pro‐adipogenic factor, which may serve as a druggable target to inhibit the turnover and accumulation of adipocytes in obese patients.Grant Funding Source: Supported by NIH Ruth L. Kirschstein National Research Service Awards for Individual Predoctoral Fellowships to Promote Diversity in Health‐Related Research
Crotalphine: fluorescent analogues, structural analysis and its effect on co‐cultures of peritoneal macrophages and DRG neurons (LB615)
Authors: Remuzgo Cesar , Sampaio Sandra , Picolo Gisele , Machini Maria , Cury Yara 
Published: June 2021

Crotalphine (CRP) is a 14‐mer peptide isolated from the venom of C. durissus terrificus “rattlesnake” that triggers long‐lasting antinociception (3‐5 days). This effect is mediated by activation of peripheral CB2 cannabinoid receptors, the subsequent release of dynorphin A and activation of opioid receptors. In inflamed tissues, granulocytes and macrophages are the main source of opioid peptides, which play a substantial role in the modulation of pain via interactions with opioid receptors located in peripheral sensory nerve fibers. In this work, we synthesized CRP and fluorescent analogues, analyzed its structure and investigated its effect on peritoneal macrophages and co‐cultures of dorsal root ganglia (DRG) neurons and peritoneal macrophages. All procedures were approved by ethics committee of the Butantan Institute (CEUAIB: 946/12) and performed in accordance to the International Association for the Study of Pain guidelines. CRP and the obtained analogues [Glu1]‐CRP, carboxyfluorescein (CF)‐[Glu1]‐CRP, CF‐[Gln1]‐CRP were able to induce antinociception. Circular dichroism spectra of CRP and [Glu1]‐CRP have shown bands indicative of the presence of random and helix structure and has allowed the construction of a 3D‐structure model of CRP. Fluorescence microscopy and mass spectra analysis of peritoneal resident or LPS‐activated macrophages treated with CF‐[Gln1]‐CRP (1 µM) showed that this peptide internalizes and is distributed in the cytoplasm of the cells, releasing dynorphin A (1‐13) and Met‐enkephalin to extracellular medium. Internalization of κ‐opioid receptors into small DRG neurons in co‐cultures of peritoneal macrophages and DRG neurons treated with bradykinin (10 μM) and CRP (1 μM), was also observed. On the other hand, we do not observed internalization of κ‐opioid receptors into macrophages. In conclusion, the data suggest a functional crosstalk between macrophages and DRG neurons in the presence of CRP. This phenomenon could contribute to the in vivo antinociceptive effect of this molecule.Grant Funding Source: Supported by FAPESP (2011/09333‐7), CeTICs/CEPID/FAPESP (2013/07467‐1) and INCTTox/CNPq (150848/2010‐2)
Further characterization of the allosteric site of action of amiodarone at Gq‐linked muscarinic receptors (LB616)
Authors: Ellis John , Jayasuriya Gihan , Elmslie Gwendolynne , Burstein Ethan 
Published: June 2021

We have previously reported that amiodarone interacts with a novel allosteric site on muscarinic receptors. Amiodarone’s most striking effect is to enhance the maximal response elicited by muscarinic agonists at the M1, M3, and M5 receptors. Furthermore, the quaternary analog N‐ethylamiodarone (NEA) is inhibitory at these receptors and appears to compete with amiodarone at that allosteric site. In the present studies, we show that dronedarone (DRON) also modulates Gq‐mediated responses, although in a more discriminating manner. For example, DRON markedly enhances pilocarpine‐stimulated release of arachidonic acid from CHO cells, via the M3 receptor subtype, but does not affect the acetylcholine‐stimulated response. In comparison to these effects at M3, DRON is strongly inhibitory toward both pilocarpine and acetylcholine at the M1 subtype. The effects of DRON are consistent with an interaction at the amiodarone site: DRON inhibits the enhancement of acetylcholine’s response produced by amiodarone at the M3 subtype; and, NEA reverses the enhancement of pilocarpine’s response at M3 produced by either DRON or amiodarone. In studies with the M1‐selective allosteric agonist AC260584, amiodarone enhanced the maximal response observed, whereas DRON was inhibitory. On the other hand, BQCA, the well‐known PAM that dramatically enhances the potency of acetylcholine at the M1 subtype, had no effect on the response profile of AC260584. In summary, DRON acts at Gq‐linked muscarinic receptors in a manner that complements amiodarone and provides an additional tool with which to investigate this novel allosteric site.Grant Funding Source: Supported by PHS R01 005214 to JE