bluuebunny/crossref_metadata_2025_split huggingface dataset:
Caustic Lesions of Esophagus in Rats and Therapy with Stable Gastric Pentadecapeptide BPC 157
Authors: Peklic Marina , Drmic Domagoj , Pavlov Katarina Horvat , Blagaic Alenka Boban , Seiwerth Sven , Sikiric Predrag 
Published: June 2021

AimWe focused on caustic lesions of the esophagus in rat and therapy with the stable gastric pentadecapeptide BPC 157. Caustic lesions of the esophagus are one of the life‐threatening morbidities after ingestion of acid or alkali, as an accidental ingestion affecting mostly children. BPC 157 is an anti‐ulcer peptide, stable in human gastric juice, and shown as a novel mediator of Robert's cytoprotection with the beneficial effect in the whole gastrointestinal tract (Curr Pharm Des. 2017;23(27):4012–4028; Curr Pharm Des. 2014;20(7):1126–35; Curr Pharm Des. 2010;16(10):1224–34). Providing a direct cytoprotective effect of BPC 157 therapy on esophagus as well (Curr Pharm Des. 2010;16(10):1224–34), we assume its beneficial effect when given after caustic substance ingestion.MethodsDeeply anesthetized Albino Wistar rats, 200 g b.w., received directly 0.5 mL of the caustic substance (10% NaOH or 20% HCl respectively) at the proximal part of the esophagus with the free passage to the stomach, and immediately thereafter, BPC 157 10 μg/kg or saline (5 mL/kg) intraperitoneally. Assessment (macro/microscopy), was at the sacrifice, at 1, 2, 5 and 10 minutes. At 1 and 10 minutes, in proximal, middle and distal esophagus tissue samples, oxidative stress assessment was by thiobarbituric acid (TBA) reactivity as malondialdehyde equivalents (MDA). Likewise, at 1 and 10 minutes post‐injury, we determined nitric oxide (NO) levels in proximal, middle and distal esophagus tissue samples using the Griess reaction (Griess Reagent System, Promega, USA).ResultsGross assessment at all time intervals revealed a marked reduction of the lesions induced by both caustics in rats that received BPC 157 therapy. Microscopic examination was along with macroscopic evaluation. Thus, BPC 157‐rats exhibited markedly less damage in the proximal, middle and distal esophagus when challenged with caustics. Of note, in addition to the extensive esophageal caustic lesions, controls exhibited a huge increase of the oxidative stress and NO‐levels following caustic application. By contrast, rats damaged by caustic that subsequently received BPC 157 therapy had markedly less MDA and NO‐tissue level, even close to those values noted in the healthy tissue samples.ConclusionBPC 157 has a cytoprotective effect on the esophagus. Likewise, of particular importance is the evidence that this beneficial effect is obtained when the BPC 157 therapy is given after caustic ingestion. Thus, a practical importance of these findings should be emphasized.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
D‐Ribose Induced NLRP3 Inflammasome Activation in Podocytes by Increase in Advanced Glycation End‐Products
Authors: Hong Jinni , Zhang Qinghua , Li Guangbi , Ritter Joseph , Li Weiwei , Wang Xuemei , Li Pinlan 
Published: June 2021

Recent studies have demonstrated that D‐ribose levels are increased in patients with type II diabetes mellitus and that increased blood D‐ribose levels are involved in the development of diabetic complications such as diabetic encephalopathy and nephropathy. However, the mechanisms mediating the pathogenic role of D‐ribose remain poorly understood. In the present study, we treated podocytes with different concentrations of D‐ribose and examined its potential effects on NLRP3 inflammasome activation and podocyte function. Using confocal microscopy, D‐ribose was found to markedly increase the co‐localization of caspase‐1 and NLRP3 by 1.8 folds compared to control, which suggests enhanced NLRP3 inflammasome formation. By ELISA, we demonstrated that in podocytes treated with D‐ribose, a major product of NLRP3 inflammasome‐IL‐1β was significantly increased by 2.05 folds, indicating the activation of NLRP3 inflammasomes. Western blot analysis further confirmed that D‐ribose increased the level of active caspase‐1 in podocytes by 2.09 folds, another parameter for activation of NLRP3 inflammasomes, which was substantially attenuated by caspase‐1 inhibitor, YvAD and ROS scavenger, NAC, respectively. In addition, D‐ribose‐induced NLRP3 inflammasome activation could be blocked by inhibitor of advanced glycation end‐products (AGEs) formation and their chemical breaker or receptor antagonist, aminoguanidine, aylthiazolium chloride and RAGE siRAGE by 43%, 50% and 73%, respectively. These results suggest that D‐ribose induces NLRP3 inflammasome activation in podocytes in an AGE‐dependent manner, which may be a triggering mechanism of diabetic nephropathy.Support or Funding InformationSupported by NIH grants DK54927 and HL075316 to Pinlan Li, NSFC grants 81573763 to Weiwei Li, and NSFC grants 81530099 to Xuemei WangThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differential inactivation mechanism and covalent adduct formation of ALDH1A1 and ALDH1A2 by WIN18,446
Authors: Zhong Guo , Zelter Alex , Davis Trisha N. , Hoopmann Michael R. , Mortiz Robert L. , MacCoss Michael , Nelson Wendel L. , Isoherranen Nina 
Published: June 2021

WIN18,446 [N,N′‐1,8‐octamethylenebis‐(2,2‐dichloroacetamide)] was discovered as a reversible male contraceptive, which inhibits spermatogenesis in many species, including humans. The contraceptive activity of WIN18,446 is believed to be due to inhibition of the biosynthesis of the critical regulator of spermatogenesis, intratesticular all‐trans‐retinoic acid (atRA), via inhibiting aldehyde dehydrogenase 1A1 (ALDH1A1) and ALDH1A2. However, the usefulness of WIN18,446 is limited due to its off‐target inhibition of ALDH2, the key enzyme eliminating acetaldehyde. Previous studies have shown that in the presence of NAD+, WIN18,446 is a reversible inhibitor of human ALDH1A1, and a time‐dependent, irreversible inhibitor of ALDH1A2. However, neither the mechanism of ALDH1A2 inactivation by WIN18,446 nor the discrepancy between ALDH1A1 and 1A2 inhibition is understood. Using Q‐TOF MS, we identified an aldehyde metabolite of WIN18,446 formed by ALDH1A2, in the presence of cofactor NAD+. In contrast, the aldehyde metabolite was formed by ALDH1A1 only in the absence of NAD+. Similarly, the inactivation of ALDH1A2 by WIN18,446 was NAD+ dependent while ALDH1A1 was only inactivated in the absence of NAD+. Adducts of WIN18,446 with ALDH1A2 were identified with shotgun proteomics and a modified version of Kojak software that allows searching of any mass modified peptides. Using the proteomic MS data, a novel WIN18,446 derived adduct of Cys320 and Cys319 of ALDH1A2 was identified with mass change of 295Da. Based on the identity of this adduct, a mechanism of ALDH1A2 inactivation by WIN18,446 and the pathway to the aldehyde metabolite was proposed (Fig. 1). Interestingly, the proposed mechanism of inactivation does not require NAD+ suggesting that NAD+ acts as an allosteric modulator affecting WIN18,446 binding orientation within the ALDH1A active site instead of contributing to the catalysis. This interpretation is consistent with existing knowledge of ALDH family enzyme structure‐function. In conclusion, using novel MS proteomics methods we have identified the inactivation mechanism of ALDH1A by WIN18,446, which provides important information to guide development of novel male contraceptive inhibitors of ALDH1A enzymes and to assess structure‐function of ALDH1A inhibitors.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Pharmacological Evaluation of Novel Chloroquine Analogues in Breast Cancer Cells
Authors: Cao Peter , Patel Dhvani , Ventura Dominic , Heakal Yasser 
Published: June 2021

Targeting autophagy, a conserved cellular process required for the degradation and recycling of proteins and cellular organelles, has emerged as a potential therapeutic approach for cancer treatment. Although, its role in cancer progression remains controversial, autophagy inhibition has been shown to be potentially beneficial in pre‐clinical studies. Constitutively active autophagy due to cellular stress has been implicated in tumor progression and therapeutic resistance. Pharmacological inhibition of autophagy using chloroquine, an approved antimalarial agent, has been investigated in preclinical cancer models and is currently being investigated in several ongoing clinical trials. However, several problems may prevent the translation of preclinical findings into the clinic such as the potential adverse events associated with chloroquine. Additionally, most preclinical studies utilize high chloroquine concentrations to inhibit autophagy which may not be achieved clinically. Therefore, we hypothesized that the design and synthesis of novel chloroquine polyamine conjugates may result in compounds that inhibit autophagy in breast cancer cells at lower concentrations due to enhanced cellular uptake through the polyamine transporter. Assessment of the biological activity of the chloroquine analogs using MTT assay demonstrates the chloroquine polyamine conjugates induce cell death at lower concentrations relative to chloroquine. Additionally assessment of the ability of these compounds to inhibits autophagy indicate that they inhibit autophagy at lower concentrations as indicated by the accumulation of LC3B‐II and p62, a protein known to undergo autophagy‐dependent degradation. Future studies will aim at investigating the mechanism of the enhanced cytotoxic activity of these compounds.Support or Funding InformationD'Youville College and D'Youville College School of PharmacyThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
PAR1 and p38 MAPK Regulation of Endothelial Pro‐Inflammatory Responses
Authors: Rada Cara , Grimsey Neil , Canto Isabel , Mejia‐pena Hilda , Trejo JoAnn 
Published: June 2021

Endothelial cells line the lumen of blood vessels and create a semi‐permeable barrier that is disrupted during vascular inflammation resulting in increased permeability, tissue edema, and subsequent organ failure. Endothelial dysfunction also results in cytokine production and leukocyte recruitment. However, the molecular mechanisms behind this pro‐inflammatory state remains to be elucidated. Protease‐activated receptor‐1 (PAR1) is a G protein‐coupled receptor for the coagulant protease thrombin and mediates hemostasis, thrombosis, and inflammatory response to vascular injury. However, the distinct mechanism of how endothelial PAR1 promotes vascular pro‐inflammatory responses represents a major gap in knowledge. We recently discovered that thrombin activation of PAR1 stimulates p38 MAPK activation through a non‐canonical pathway mediated by TAB1 to promote endothelial permeability. PAR1‐induced p38 activation has been linked to vascular barrier disruption; however, the mechanism as to how p38 may be integrated into major signaling pathways that promote pro‐inflammatory responses is not known. We found that thrombin‐stimulated p38 signals independent of the classic g‐protein pathways of RhoA activation and MLC phosphorylation to induce endothelial barrier permeability. Instead, thrombin‐stimulated p38 appears to be signaling through phosphorylation of the MK2 and HSP27. We further show MK2 activity is PAR1 and p38 dependent and regulates differential phosphorylation of its direct substrate, HSP27. Additionally we show HSP27 activity is p38 and PAR1‐dependent. The p38‐MK2‐HSP27 signaling pathway also appears to modulate HUVEC monolayer permeability. These studies show thrombin‐induced p38 signals through MK2 and HSP27 cascade to regulate endothelial barrier permeability.Support or Funding InformationT32 Pharmocological Sciences Training Grant (NIGMS T32 GM007752) (C. Rada) NIH NIGM R01 GM116597 (J. Trejo)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Novel PRKAG2 Mutation (K475E): Early‐Onset Cardiac Phenotype and Targeted Therapy
Authors: XU YANCHUN , Liu Hong , Phornphutkul Chanika , Hardie Grahame , Dudley Samuel C. , Padbury James F. , Tseng Yi‐Tang 
Published: June 2021

BackgroundMutations in PRKAG2 have been implicated in hypertrophic cardiomyopathy (HCM). We identified a novel PRKAG2 mutation, K475E, in a neonate with prenatal onset of HCM noted on a 27‐week prenatal ultrasound. We have demonstrated in vitro that K475E mutation led to increases in cardiomyocytes size and area which can be reversed by rapamycin.ObjectiveTo characterize K475E‐induced cardiac phenotype in vivo, we developed two new transgenic mouse lines with cardiac‐specific expression of the human wild type (TgWT) or K475E mutant (TgK475E) PRKAG2. The goal of this study was to explore the effectiveness of rapamycin for targeted therapy for PRKAG2 mutation‐induced HCM.MethodsHuman WT and K475E DNA constructs were inserted into a pBluescript vector with the mouse α‐myosin heavy chain promoter. Transgenic mice were established by pronucleus microinjection. Rapamycin was treated from two‐week to six‐aged mice. Cardiac hypertrophy was examined and cardiac function changes were monitored by echocardiography.ResultsHypertrophy in TgK475E mice was confirmed by their significant increases in heart weight/body weight and heart weight/tibial length ratios versus the non‐transgenic control and TgWT mice. Histology revealed large vacuoles and high glycogen content in TgK475E myocytes throughout the ventricles. Echocardiography confirmed that LV mass dramatically increased in TgK475E mice starting from 5 weeks of age. Rapamycin treatment dramatically decreased 1) the ratios of heart weight/body weight and heart weight/tibia length and 2) glycogen overload in TgK475E mice. Moreover, K475E mutation caused inhibition of AMPK which was rectified by rapamycin treatment.ConclusionThese results indicate the transgenic mouse model reflects the early onset phenotype of the K475E mutation‐induced HCM. The mTOR signaling pathway is involved in K475E induced HCM. Our study suggests that mTOR inhibition strategy as targeted therapy for PRKAG2 induced HCM is effective and should be studied further.Support or Funding Information1. NIH P30GM114750 2. Oh‐Zopfi Research Award, Women & Infant's Hospital, RI.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Anti‐inflammatory effect of a new formulation: Palmitoylethanolamide/Baicalein in a murine model of Myocardial Ischaemia/Reperfusion Injury
Authors: Cuzzocrea Salvatore , D'Amico Ramona , Fusco Roberta , Gugliandolo Enrico , Crupi Rosalia , Impellizzeri Daniela , Di Paola Rosanna 
Published: June 2021

Myocardial ischemia/reperfusion (I/R) injury is the principal cause of death, happens after prolonged occlusion of the coronary arteries. The first intervention to limit myocardial damage is directed toward restoration of perfusion as soon as possible, however infarction triggers a powerful inflammatory response and a significant oxidative stress.Palmitoylethanolamide (PEA), is a well‐known fatty acid amide‐signaling molecule that possess an important anti‐inflammatory and analgesic effects. Nevertheless, PEA does not possess the ability to prevent free radicals formation. Baicalein, a bioactive component isolated from a Chinese herbal medicine, has multiple pharmacological activities, such as a strong anti‐oxidative effects. A combination of PEA and Baicalein could have beneficial effects on oxidative stress produced by inflammatory response. In the present study we investigated the effects of composite containing PEA and Baicalein in a model of myocardial I/R injury. Myocardial ischemia/reperfusion injury was induced by occlusion of the left anterior descending coronary artery for 30 min followed by 2 h of reperfusion. PEA‐Baicalein (10:1), was administered (10 mg/kg) 5 min before the end of ischemia and 1 h after reperfusion. In this study, we clearly demonstrated that PEA‐Baicalein treatment reduces myocardial tissue injury, neutrophils infiltration, markers for mast cell activation expression such as chymase and tryptase and pro‐inflammatory cytokines production (TNF‐α, IL‐1β). Moreover, PEA‐Baicalein treatment reduces nitrotyrosine and PAR formation, inhibits NF‐kB nuclear translocation and modulates apoptosis pathways.Accordingly, these results support the idea that the association between PEA and Baicalein should be further studied as a potent candidate for the treatment of myocardial I/R injury.Support or Funding Informationno fundingThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
AT1R Dependence of Proangiogenic Ang‐(1–7)/Mas Signaling in Endothelial Cells and Endothelial Progenitors
Authors: Exner Eric Charles , Stodola Timothy J. , Hoffmann Brian R. , Greene Andrew S. 
Published: June 2021

The renin‐angiotensin system (RAS) is a critical regulator of microvascular function and cardiovascular risk. Success of clinical interventions that interfere with the signaling of angiotensin II (AngII) at the angiotensin II receptor type 1 (AT1R) suggested signaling via AT1R is predominantly associated with pathology; however, recent studies have reaffirmed that AngII/AT1R signaling has a key role in maintaining vascular homeostasis. Angiotensin‐(1–7) (Ang‐(1–7)) is a RAS effector peptide that counteracts many effects of pathologically high AngII. In a previous study, we observed that at low doses, both AngII/AT1R and Ang‐(1–7) signaling through the receptor Mas restore angiogenesis in a low renin model of microvascular dysfunction. We have also observed beneficial effects of low dose AngII on endothelial progenitor cell (EPC) function. While competitive inhibition of ligand binding at the converse receptor did not abolish the effects of each peptide, AT1R was identified as a component of the Ang‐(1–7)/Mas signaling complex. It has been suggested that AT1R and Mas form heterodimers based on data showing rescued function of mutant AT1R upon coexpression of Mas. Here, we test the hypothesis that Ang‐(1–7) has proangiogenic effects in both endothelial cells (ECs) and EPCs that is dependent upon the presence of AT1R. In the Dahl salt‐sensitive (SS) rat, a low‐renin model of microvascular dysfunction, we confirm the beneficial effects of low‐dose Ang‐(1–7) on angiogenesis and observe Ang‐(1–7) mediated restoration of EPC function. In an AT1R knockout on the SS background, the ability of Ang‐(1–7) to restore angiogenesis and EPC function is lost. In addition, in vitro analogues of angiogenic capability recapitulate this phenotype in both ECs and EPCs. Using qPCR arrays and mass‐spectrometry based proteomics, we find both the transcriptional response and the Ang‐(1–7)/Mas signaling complex to be altered in the absence of AT1R. These results support the hypothesis that AT1R is a key component of the Ang‐(1–7)/Mas signaling pathway and further support the previous hypothesis by others and ourselves that AT1R and Mas heterodimerize. Computational models of AT1R and Mas have been constructed to examine the potential mechanism of AT1R and Mas heterodimerization, with biological confirmation to follow.Support or Funding InformationThis work was supported by NHLBI grant F30HL131153.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Neoadjuvant Chemotherapy Decreases Angiogenesis Potential and Microvascular Function in Human Breast Cancer Patients
Authors: Norwood Toro Laura E. , Linn Jasmine , Hockenberry Joseph C. , Kong Amanda L. , Flister Michael J. , Beyer Andreas M. 
Published: June 2021

RationalChemotherapy (CT) is a necessary treatment to prevent the growth and survival of cancer cells; however, CT is well established to have adverse impact on the cardiovascular (CV) system even years after cessation of treatment. Much work has been dedicated to studying the effects of CT drugs on the tumor vasculature, but little evidence exists showing the effects on the host microcirculation. Microvascular (MV) dysfunction is a known early indicator of numerous CV disease phenotypes, including heart failure. The goal of this study was to evaluate whether previous CT treatment has any lasting effect on the human MV.MethodsTo study the effect of CT on the MV, flow mediated dilation (FMD) was evaluated in freshly isolated adipose microvessels from cancer patients with and without previous CT. Angiogenic potential was assessed using matrigel plug assays with samples isolated from human adipose. Effect of CT on endothelial cell (EC) and cancer cell survival was determined using MTT; EC permeability was established using FITC‐dextran in transwells with a EC monolayer.ResultsIn cultured cells doxorubicin (dox; 100 nM) causes a decrease in MTT in human cancer, but not EC. Ex vivo treatment of the vessels with dox reduced nitric oxide (NO) mediated dilation by 90% compared to untreated controls (A). Similarly, MV FMD was severely decreased in vessels from patients with neoadjuvant CT (cessation of therapy ~ four weeks prior) compared to untreated control subjects (B). Smooth muscle dependent dilation to papaverine was not affected in either study (C). Using an ex vivo matrigel assay, we found a significant decrease in angiogenesis potential in samples treated with dox (D). In line with these findings, dox causes a decrease in EC cell permeability, suggesting a potential contributing mechanism.ConclusionsDox in clinically relevant doses causes increased lethality in cancer cells versus EC, but induces MV dysfunction, decreased angiogenic potential, and elevated EC permeability. Our studies show for the first time the effect of dox on human MV function and may lead to ways to prevent adverse CV side effects from CT without affecting cancer cell lethality.Support or Funding InformationThis work was supported by NIH R01 HL133029, We Care Foundation Grant; MCW‐Cardiovascular Center Pre‐PPG grant, Advancing a Healthier Wisconsin – Redox Biology Grant.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.