bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human Stem Cell Derived Cardiomyocyte Maturation is Regulated by Glucose Levels and Metabolic Hormone Supplementation
Authors: Klug Heather May , Karbassi Elaheh , Yang Xiulan , Pabon Lil , Murry Charles 
Published: June 2021

Methods to differentiate stem cells into cardiomyocytes have been well established. However, a limitation for the successful application of these cells for research and medicine has been their fetal‐like phenotype with respect to cell size, contractility, calcium handling, metabolism, and electrophysiology. We sought to increase the maturity of human pluripotent stem cell derived cardiomyocytes (hPSC‐CMs) through metabolic pathway regulation. We hypothesized that switching the main metabolic substrates from glucose to fatty acids, mimicking the switch from placental to breast milk nutrient consumption that occurs during development, will increase hPSC‐CM maturation. RUES2 embryonic stem cells were differentiated into cardiomyocytes and then treated with base media with varying glucose and calcium concentrations and fatty acid supplementation. Using qRT‐PCR, we measured an inverse relationship between glucose levels and markers of cardiomyocyte maturation: increased expression of cardiac troponin I relative to skeletal troponin I isoforms (TNNI3:TNNI1), increased expression of metabolic (CPT1B, PPARGC1A) and electrical maturity markers (KCNJ2, RYR2). These maturation markers were not influenced with fatty acid supplementation but were enhanced upon the addition of thyroid hormone and dexamethasone. To understand the mechanisms by which media nutrients and signaling molecules cause phenotypic changes we investigated the relationship between maturation and global epigenetic state. Western blot revealed increases in global acetylation levels, measured by histone H3 acetylation, linked to maturation signaling. These findings depict a direct relationship between glucose metabolism and the development of a mature phenotype in hPSC‐CMs, mediated by epigenetic mechanisms.Support or Funding InformationWashington Research Foundation Undergraduate FellowshipThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Toxoplasma gondii alters NMDAR signaling and induces signs of Alzheimer's disease in wild‐type, C57BL/6 mice
Authors: Torres Luisa , Bynoe Margaret S 
Published: June 2021

Alzheimer's disease (AD) is a progressive neurodegenerative disease associated with cognitive decline, resulting in a complete loss of basic functions. The ubiquitous apicomplexan parasite Toxoplasma gondii (T. gondii) infects up to one third of the world's population and is implicated in AD. We infected C57BL/6 wild‐type male and female mice with 10 T. gondii ME49 cysts and assessed whether infection led to behavioral and anatomical effects using immunohistochemistry, immunofluorescence, Western blotting, cell culture assays, as well as an array of mouse behavior tests. We previously reported that T. gondii infection induced two major hallmarks of AD in the brains of C57BL/6 male and female mice: beta‐amyloid (Aβ) immunoreactivity and hyperphosphorylated Tau. Infected mice showed significant neuronal death, loss of N‐methyl‐D‐aspartate receptor (NMDAR) expression, and loss of olfactory sensory neurons. T. gondii infection also caused anxiety‐like behavior, altered recognition of social novelty, altered spatial memory, and reduced olfactory sensitivity. We have new data that suggests that T. gondii might be recruiting the NMDA receptor to the parasitophorous vacuole, a structure that protects T. gondii from the phagolysosomes of its host. This indicates that T. gondii might use the NMDA receptor of the host to survive and might explain the reduction of NMDAR that we observed with T. gondii infection. Since reduced NMDAR expression coincides with Aβ immunoreactivity, it is possible that T. gondii infection leads to reduced NMDAR expression which then leads to Aβ immunoreactivity.Support or Funding InformationThis work was supported by NIH grant R01 NS063011 to M.S BynoeThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protease‐Activated Receptor 2 Drives Colonic Epithelial Wound Healing via EGFR Transactivation
Authors: Bandara Mahesha N.S. , MacNaughton Wallace Keith 
Published: June 2021

BackgroundCompromised intestinal barrier function is a prominent feature of inflammatory bowel diseases. Epithelial restitution is required in the resolution of inflammation. For this to occur epithelial cells need to transition to migratory phenotype from its normal barrier phenotype. The inflammatory milieu of the intestinal epithelium contains a variety proteases which are upregulated in IBD patients and which are agonists of protease activated receptors (PARs). PAR2 is membrane‐spanning cell surface receptor which is widely present in the gastrointestinal tract. PAR2 activation can induce expression of cyclooxygenase‐2 (COX‐2) and transactivation of epidermal growth factor receptor (EGFR) in the intestinal epithelium. However, the specific roles of PAR2 activation in IBD remain unclear.HypothesisWe hypothesized that PAR2 activation drives COX‐2 and EGFR dependent epithelial wound healing.MethodsCMT93 mouse intestinal epithelial cells were grown to confluence. PAR2 mRNA expression was detected by RT‐PCR and confirmed by cDNA sequencing. PAR2 A5 antibody was used to detect total PAR2 by immunocytochemistry and the tight junction protein, ZO‐1, demarcated apical and basolateral surfaces. PAR2 was activated by selective activating peptide 2‐furoyl‐LIGRLO‐NH2 (2fLI). Functionality of PAR2 was tested by measuring intracellular Ca2+, since it has been shown that PAR2 activation elicits Ca2+‐dependent signaling. Scratch wounds were made in post‐confluent CMT93 monolayers and imaged using the IncuCyte™ live‐cell imaging system over 24 hours. COX‐2 protein was measured by western blotting and inhibited in wound healing assays with NS‐398 (10 μM). EGFR was inhibited by the EGFR tyrosine kinase inhibitor PD153035 (10 nM) and activated using human EGF (10 ng/ml and 5 ng/ml).ResultsImmunocytochemistry revealed that PAR2 is expressed on mostly basolateral membranes with some expression on apical membrane and punctate immunoreactivity in the cytoplasm in polarized cells. Concentration‐response studies revealed that 2.5 μM 2fLI elicited the highest intracellular Ca2+ response (EC50 = 0.5 μM). PAR2 activation at 10 μM 2fLI significantly increased wound healing (P≤ 0.0001). However, PAR2 activation by 2fLI did not induce COX‐2 expression, nor did COX‐2 inhibition alter PAR2‐induced wound healing. PAR2‐induced wound healing was inhibited by the EGFR tyrosine kinase inhibitor (P ≤ 0.001). Moreover, 2fLI induced wound healing is comparable to the that of EGF (5 ng/ml).ConclusionPAR2 activation drives wound healing in CMT93 epithelial cells through the transactivation of EGFR and not through induction of COX‐2. This study enhances our understanding of how proteases in the inflammatory milieu may drive epithelial restitution and mucosal healing.Support or Funding InformationCIHRThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mitochondrial derived reactive oxygen species activates NLRP3 inflammasome in cardiac fibroblasts in sepsis
Authors: Yan Jun , Zhang Hao , Wang Hao , Tao Aibin , Rui Tao 
Published: June 2021

BackgroundSepsis is a common disease in intensive care unit in hospital. It is caused by exaggerated host response to infection. Septic myocardial dysfunction plays an important role in pathogenesis of multiple organ failure and subsequent high mortality of patients with sepsis. Our previous studies indicate that activation of NLRP3 inflammasome in cardiac fibroblasts (CF) contributes to sepsis‐induced myocardial dysfunction. The aim of the present study is to identify the role of mitochondrial ROS (mtROS) in the activation of the NLRP3 inflammasome in CF.MethodsCultured CF was primed with LPS (1 ng/mL) followed by ATP (3 mmol/L) treatment for activation of NLRP3 inflammasome. CF mtROS was detected with MitoSOX reagent, a probe specifically detects mitochondrial ROS. NLRP3 inflammasome activation was assessed with Western blot, ELISA and co‐immunoprecipitation.ResultsATP challenge of CF primed with LPS increased CF mtROS production which was attenuated by pretreatment of the LPS primed CF with mito‐TEMPO (25 mmol/L), an ROS scavenger targeted at mtROS. ATP challenge of the LPS primed CF activated the NLRP3 inflammasome as indicated by increase in intracellular levels of NLRP3, pro‐IL‐1β and released IL‐1β. Mito‐TEMPO pretreatment of the CF with LPS+ATP prevented the increase of released IL‐1β in CF. Finally, ATP challenge of the LPS primed CF promoted NLRP3 interaction with the ASC. The Mito‐TEMPO pretreatment of the CF diminished the interaction between the NLRP3 and ASC.ConclusionOur data suggest that mitochondrial derived ROS plays a pivotal role in the activation of the CF NLRP3 inflammasome in sepsis.Support or Funding InformationThe study was supported by Natural Science Foundation of Jiangsu Province (BK20151332) and Natural Science Foundation of China (81370333).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Actions of Angeli's salt, a nitroxyl (HNO) donor, on Trypanosoma cruzi infection
Authors: Pinge‐Filho Phileno , Pereira Rito Santo , Malvezi Aparecida Donizette , Lucchetti Bruno Fernando Cruz , Cecchini Rubens , Yamauchi Lucy Megumi , Yamada‐Ogatta Sueli Fumie , Verri Waldiceu Aparecido , Martins‐Pinge Marli Cardoso , Tatakihara Vera Lucia Hideko 
Published: June 2021

Chagas disease, caused by Trypanosoma cruzi infection, is endemic to much of Latin America, but also present in the United States (U.S.) The goal of treatment is to eliminate the parasite and decrease the probability of cardiomyopathy and interrupt the cycle of disease transmission. Currently, only benznidazole (BZN) and nifurtimox are recognized by the World Health Organization as effective drugs for treatment of Chagas disease, but both present low cure rates in the chronic phase and often have serious side effects. T. cruzi infection produces an intense inflammatory response in several tissues which is critical for controlling parasites proliferation and evolution of Chagas disease. Compounds liberating nitric oxide (NO donors) have been used as therapeutic agents against T. cruzi. Here we test HNO donor, Angeli's salt (AS) in the outcome of T. cruzi infection. C57BL/6 mice were infected with T. cruzi (Y strain, 5 × 103 trypomastigotes) intraperitoneally (ip). After 15 minutes of infection and the subsequent 12 days, the animals were treated with different concentrations of AS ip. The control group received 100 uL of PBS by the same route. Treatment with AS showed decreased parasitemia and in the number of nests in the cardiac tissue, decreased oxidative stress in erythrocytes and improved leukopenia and thrombocytopenia resulting in reduced disease severity. The in vitro treatment was able to reduce the parasite internalization rate and release of infective trypomastigotes by these cells. Our results suggest for the first time the therapeutic potential of AS in a mouse model of Chagas disease.Support or Funding InformationCAPES, Fundação Araucária, CNPQ, UniZambeze and MCTESTP Mozambique.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Association of Factors Involved in the Pathogenesis of Chagas Disease
Authors: Hagström Luciana , Wesley Moisés , Fereira Tatiana Shiroma Borges , de Souza Alves Jehny Pereira , de Cássia Rosa Ana , Dallago Bruno , Nitz Nadjar , Hecht Mariana Machado 
Published: June 2021

Chagas disease, caused by the protozoan Trypanosoma cruzi, represents a serious health public problem with around seven million infected people worldwide. Approximately 30% of infected individuals will develop cardiac disorders during the chronic phase. However, the pathogenesis of Chagas disease remain incompletely elucidated with fundamental questions partially unanswered as the factors that lead to the development of clinical manifestations. Thereby, this study aims to investigate the importance of the association of T. cruzi strain, host sex, host immunological factors and changes in heart tissue in the progression of cardiac lesions in experimental Chagas disease. For this, 60 BALB/c mice were divided into three groups according to the T. cruzi strain (discrete typing unit TcI, TcII and TcVI). They were analyzed in acute phase (30 days pos‐infection ‐ 30dpi) and chronic phase (90 dpi). Ten uninfected mice formed the control group. Each group were formed by five males and five females. After the experimental period, mice were euthanized and blood and heart were collected. Anti‐T cruzi antibody and anti‐cardiac tissue antibody production were evaluated by ELISA. Parasite load in the heart was determined by qPCR. Histopathological analysis of heart was also performed. Most of the animals produced anti‐T. cruzi IgM antibody in the acute phase, whereas all mice were seropositive for IgG in chronic phase. TcI was characterized by elevated antibody levels with autoantibodies titers against the heart in chronic phase significantly higher than the other groups. Besides that TcI presented a greater number of parasites both in acute and chronic phase as well as more damage in heart tissue. No difference was observed between males and females. Amastigote nests were seen in mice analyzed in acute phase in TcI and TcVI, but not in TcII. The results confirm the participation of autoimmune response in chronic Chagas disease, which varies according to T. cruzi strain. It seems that the TcI was the most aggressive. However, the relationship between the studied factors and the severity of disease has not been established.Support or Funding InformationThis study was financed by Fundação de Apoio a Pesquisa do Distrito Federal (FAPDF), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Different Roles of Epigenetic Regulators and Inflammasome in Hepatocellular Carcinoma Tumorigenesis in Patients with ASH vs. NASH
Authors: Jia Yue , Ji Ping , Nguyen Luan , French Barbara , Tillman Brittany , French Sameul W 
Published: June 2021

As the fifth most common cancer and the second leading cause of cancer related deaths worldwide, hepatocellular carcinoma (HCC) is an aggressive tumor type with poor prognosis causing 250,000 to one million deaths annually. Alcoholic steatohepatitis (ASH) and non‐alcoholic steatohepatitis (NASH) are the two major growing risk factors and both may develop liver fibrosis or even HCC. However, compared with the rate of patients with ASH progressing to HCC annually, it is much lower in NASH patients. The present study is to clarify the protein expression of epigenetic regulators and Inflammasome components in the oncogenesis pathway between ASH and NASH. By using the immunofluorescence method and morphometrically quantitating the fluorescence intensity in liver biopsied specimens from NASH and ASH patients, we studied the protein expression within hepatocytes cytoplasm of candidate epigenetic regulators G9A and LHPP and inflammasome component ASC. Compared with the control group patients, the expression levels of all three proteins were upregulated in the ASH and NASH group of patients (p < 0.001 in all molecules). While compared with the ASH group of patients, only the expression levels of G9A were significantly lower in the NASH group of patients (p < 0.01). The other two molecules, LHPP and ASC did not change. These results are consistent with our previous work that there are significant differences of many molecules including epigenetic modulators and the inflammasome pathway in both NASH and ASH compared to the control group. Thus, we conclude that there are significantly different molecules and pathways involved during the pathogenesis of HCC development in NASH compared to ASH which may focus the light on developing prevention methods and targeted treatments in NASH and ASH patients.Support or Funding InformationThis study was funded by NIH/AAA grant # UO‐21898‐05.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Combined effects of phenmetrazine and nor‐binaltorphimine on dopamine function and motivation to self‐administer cocaine
Authors: Estave Paige , Jones Sara 
Published: June 2021

Despite numerous clinical trials, the FDA has not approved any medication assisted treatment for cocaine addiction. Our laboratory has shown that chronic cocaine self‐administration reduced overall dopamine (DA) neurotransmission, resulting in a hypodopaminergic state. Other studies have shown that repeated, non‐contingent cocaine exposure resulted in a hyper‐functioning kappa opioid receptor (KOR) system. Thus, this study aims to examine the effectiveness of a combination therapy, targeting the dopamine transporter and the KOR, in reducing cocaine‐seeking behaviors in rats chronically exposed to cocaine.In order to understand the impacts of cocaine self‐administration on the DA and KOR systems, we used ex vivo fast scan cyclic voltammetry in the nucleus accumbens core to examine DA dynamics after chronic cocaine exposure (40 infusions of cocaine for 5 consecutive days). We found that cocaine exposure reduced stimulated DA release and attenuated cocaine potency at the dopamine transporter. Additionally, we examined KOR function through activation of KORs using the agonist U50,488, which inhibited DA release. We observed that responses to U50,488 were augmented post‐cocaine exposure.Male Sprague Dawley rats were used to determine the individual and combined behavioral effects of the DA releaser phenmetrazine and the KOR antagonist nor‐binaltorphimine (nor‐BNI). After rats self‐administered forty infusions of cocaine (1.5 mg/kg/infusion) on a fixed‐ratio 1 schedule for five consecutive days, a progressive ratio schedule was used to determine cocaine breakpoints (0.1875 mg/kg/infusion) at baseline, and following phenmetrazine (25 mg/kg/day; osmotic mini‐pump), nor‐BNI (10 mg/kg; i.p.), or a combination of the two drugs. Administration of phenmetrazine alone significantly reduced breakpoints (60% of baseline), while nor‐BNI had no effect as a monotherapy. Interestingly, cocaine breakpoints were further attenuated by the combination (20% of baseline), indicating augmented effects of targeting both the DA and KOR systems.In this study, voltammetry results showed that chronic cocaine exposure reduced DA transmission and augmented KOR function. Dual targeting of the DA and KOR systems through a combination therapy approach revealed a greater decrease in motivation to take cocaine compared to either monotherapy. Taken together, these data support the potential beneficial effects of the dopamine transporter and KOR as dual‐cellular targets to treat multiple aspects of the withdrawal syndrome, ultimately decreasing the risk of relapse in individuals with cocaine dependence.Support or Funding InformationSupport for this work was provided by R01 DA014030, P50 DA006634, and T32 DA041329This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Gut‐Brain Actions Underlying Comorbid Neuropsychiatric Disturb Associated with Inflammatory Bowel Disease
Authors: Cordaro Marika , Siracusa Rosalba , D'Amico Ramona , Gugliandolo Enrico , Peritore Alessio Filippo , Crupi Rosalia , Impellizzeri Daniela , Cuzzocrea Salvatore , Di Paola Rosanna 
Published: June 2021

Inflammatory bowel disease (IBD) is a chronic relapsing and remitting disorder characterized by inflammation of the gastrointestinal tract and its associated with different neurological and psychiatric disorders, which are integrated among the extra‐intestinal manifestation of this disease.In fact, epidemiological studies have shown an increased incidence of depression in patients with colitis, in which strong inflammation in the gut occurs. The possible mechanisms connecting gut inflammation and neuroinflammation in the brain could be via an increase in peripheral cytokines or via damaged nerve terminals in the gut.Recent evidence demonstrated that the gut‐brain‐axis has a central function in the perpetuation of IBS, and, for this reason it can be considered as a possible therapeutic target.N‐Palmitoylethanolamine‐oxazoline (PEA‐OXA) possesses anti‐inflammatory and potent neuroprotective effects. Although, recent studies have explained the neuroprotective properties of PEA‐OXA, nothing is known about its effects on gut‐brain axis during colitis.The aim of this study is to explore the mechanism and the effect of PEA‐OXA on gut brain axis in mice subjected to experimental colitis induced by intracolonic administration of Dextran sulfate sodium. Daily orally administration of PEA‐OXA 10 mg/kg daily o.s. was able to decrease body weight loss, macroscopic score and colon length and histology alteration after DSS induction. On the other hand was able to reduce histology modification and c‐FOS expression in the hippocampus.Additionally, we demonstrate that PEA‐OXA administration significantly increase neurogenesis and promoting neurotrophic grow factor release and decrease astroglial and microglial activation DSS‐induced. Moreover, PEA‐OXA, was able to decrease inflammatory marker in both hippocampus and colon.In conclusion, in our work, we demonstrate for the fort time the action of PEAOXA on gut‐brain axis during in a model of DSS induced colitis. induced. Moreover, PEA‐OXA, was able to decrease inflammatory marker in both hippocampus and colon. In conclusion, in our work, we demonstrate for the fort time the action of PEAOXA on gut‐brain axis during in a model of DSS induced colitis.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.