bluuebunny/crossref_metadata_2025_split huggingface dataset:
Axonal pathology in hPSC‐based models of Parkinson's Disease results from loss of Nrf2‐transcriptional activity at the Map1b gene locus
Authors: Ryan Tammy , Drolet Jennifer , Sherriff Keith , Coackley Carla C. , Ryan Scott D. 
Published: June 2021

IntroductionNeurological dysfunction in Parkinson's Disease (PD) is highly associated with pathological deposits of aggregated α‐synulein (α‐syn) in neurons of the brain, and yet an understanding of how early α‐syn deposits bring about neuronal pathology remains elusive. While mutations in the SNCA gene (a‐syn) are causal in rare familial forms of PD, the prevalence of α‐syn aggregates in the cortices of sporadic disease cases emphasizes the need to understand the link between α‐syn accumulation and axonal neuropathology.ApproachThis study exploits recent advances in isogenic stem cell technology to model PD using human pluripotent stem cell (hPSC) models of the familial disease. By employing 1) a combination hPSCs that harbor the SNCA‐A53T mutation contrasted against isogenic controls and 2) substantia nigra from PD patients analyzed post‐mortem, we performed gene expression profiling in search of transcriptional hubs that are disrupted early in disease pathogenesis and remain altered as the disease progresses. We then evaluated the relationship between differentially expressed genes and axonal pathology.ResultsWe show that the early accumulation of α‐syn in SNCA‐A53T hNs results in changes in gene expression consistent with the expression profile of substantia nigra from PD patients, analyzed post‐mortem. Differentially expressed genes from both PD‐patient substantia nigra and SNCA‐A53T hNs clustered to genes with regulatory motifs transcriptionally activated by the antioxidant response pathway, particularly, Nrf2 gene targets. Differentially expressed gene targets were also enriched for gene ontologies related to microtubule binding processes. We thus assessed the relationship between Nrf2‐mediated gene expression and axonal pathology in SNCA‐A53T hNs. Here, we provide direct evidence that axonal pathology in PD neurons results from impaired Nrf2‐activity on Antioxidant Response Elements (ARE) localized to a microtubule associated protein (Map1b) gene enhancer. Moreover, we show that forced activation of Map1B, rescues axonal pathology in PD neurons.SignificanceThis is the first demonstration of Nrf2‐mediated modulation of cytoskeletal elements as a potential therapy against PD‐induced axonal pathology.Support or Funding InformationThis work was supported in part by the Parkinson Society of Canada (2014‐685 to SDR), the Natural Sciences and Engineering Research Council of Canada (RG060805 to SDR)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ANATOMY BOARD GAMES PROMOTE SIGNIFICANT LEARNING ABOUT THE CARDIOVASCULAR SYSTEM
Authors: FREITAS FRANCISCO ORLANDO RAFAEL , Barreto João Erivan Façanha , de Sousa Ramos Erirlane Aquino , de Freitas Ana Paula Fragoso , de Lucena Jalles Dantas , Scafuri Ariel Gustavo , Cerqueira Gilberto Santos 
Published: June 2021

Playfulness is an important tool in the process of teaching and learning anatomy. The playful is an environment that allows pleasure, play fun and playfulness thus promoting meaningful learning. Based on these premises the objective of this work was to investigate the role of playful board games in the teaching and learning process of anatomy and physiology of the cardiovascular system. A descriptive exploratory study with a quantitative approach was carried out with the group of 136 students of both sexes. A validated standard questionnaire on anatomy and physiology of the cardiovascular system was used as a pre‐test. After the pre‐test the students used an educational board game about anatomy and physiology of the cardiovascular system composed of board, dice, hourglass and charts with anatomical and physiological questions about the cardiovascular system. After the game, a post‐test questionnaire was applied. For statistical analysis, the Mann Whitney test was considered significant when they presented a p <0.05. The mean age of the interviewees was 18.23 ± 0.73 for males and 18.13 ± 0.43 for females. He observed a statistically significant difference in the students' learning after the application of the playful educational game, that is, after applying the game the students presented a greater number of answers in the questions involving the knowledge of anatomy and physiology of the cardiovascular system. In this way, it was found that playful educational games promote significant learning as well as stimulate creativity, attention, memory, among other abilities, functioning as an important tool for teaching and learning the anatomy and physiology of the cardiovascular systemSupport or Funding InformationUFPIThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of Retinoic Acid Signaling in Epicardial‐Related Events During Late Heart Development
Authors: Moise Alexander R. , Wang Suya R. , Yu Jianshi R. , Kane Maureen R. , Xavier‐Neto Jose 
Published: June 2021

BackgroundDuring the final stages of heart development, the myocardium grows and becomes vascularized with the aid of paracrine factors and cell progenitors derived from the epicardium. Here, we study the involvement of retinoic acid (RA), a metabolite of vitamin A, in epicardial‐related developmental processes involving the formation of the coronary vasculature and the growth of the ventricular compact zone.MethodsTo explore the developmental consequences of altered RA signaling in late cardiogenic events, we employed a previously characterized model of embryonic RA excess based on mouse embryos deficient in the retinaldehyde reductase DHRS3. We also employed a complementary model based on embryonic RA deficiency generated by administering WIN 18,446, a bis(dichloroacetyl)diamine inhibitor of RA synthesis to pregnant dams. Using these two models we studied the formation of the coronary plexus and the formation and recruitment of epicardial‐derived mural cells by coronary vessels. We further explored the effect of altered RA in vitro by studying the epicardial epithelial‐to‐mesenchymal transition (EMT), and the formation, migration and differentiation of epicardial‐derived precursor cells (EPDCs).ResultsOur results demonstrated that both excess and deficiency of embryonic RA‐signaling led to a thin ventricular myocardium and aberrant coronary vessel formation and remodeling. In the case of both excess, or deficient RA‐signaling, the endothelial tubes did not become invested with epicardial‐derived vascular smooth muscle cells (VSMCs). In vitro assays of epicardial EMT combined with transcriptomic and proteomic analyses revealed that RA signaling not only affects the differentiation of EPDCs, but also controls the cytoskeletal reorganization and secretion of angiogenic and cardiotrophic factors by the epicardium.ConclusionWe report that RA signaling affects epicardial signaling and plays critical roles in the development of the myocardium and the coronary vasculature.Support or Funding InformationThis work was supported in part by the grant R01HD077260 (ARM, MAK) from the National Institutes of Health and by a Fulbright U.S. Scholar Award to ARM funded by the U.S. Department of State's Bureau of Educational and Cultural Affairs, the Commission for Educational Exchange between the United States of America and Brazil and CAPES, the Brazilian Federal Agency for Support and Evaluation of Graduate Education. Additional support to MAK was provided by the University of Maryland School of Pharmacy Mass Spectrometry Center (SOP1841‐IQB2014) and by startup funds to ARM from the Northern Ontario School of Medicine. Conflicts of interest: none.
Determining the Enhancer Proteomes in Primary Cells and Native Tissue
Authors: Steger David , Cohen Daniel , Remsberg Jarrett , Sidoli Simone , Garcia Benjamin 
Published: June 2021

Enhancers play a critical role for gene expression, yet their cell‐type‐specific establishment and maintenance remains incompletely understood. To investigate this, we surveyed the proteomic composition of enhancers in naive mesenchymal stem cells, differentiating adipocytes, and liver tissue using ChIP with selective isolation of chromatin‐associated proteins (ChIP‐SICAP). ChIP‐SICAP revealed that H3K27ac‐marked chromatin is enriched for over 500 proteins, including sequence‐specific transcription factors (TFs), chromatin remodeling complexes, and histone variants that collectively define the landscape of active enhancers in differentiating adipocytes and hepatocytes. Moreover, we observe a pivotal role for CEBP TFs at enhancers in both settings. To gain further insight into the roles of CEBPs, we performed ChIP‐SICAP for CEBPb and identified a subset of the proteins comprising the H3K27ac enhancer proteome. Although CEBPb‐bound chromatin is commonly enriched for the nuclear receptor GR and PAR‐family bZip proteins, other co‐localizing TFs, such as TEADs and HNF1a, show strict cell‐type‐specificity. Moreover, we observe that chromatin remodelers and transcriptional co‐activators such as BRD2 and MED8 are selectively enriched at sites of CEBP occupancy in response to adipogenic stimuli, suggesting these factors may play important roles with CEBPs to establish active chromatin during cell differentiation. Collectively, these observations reveal both constitutive and tissue‐specific determinants of the enhancer landscape, and provide new insights into mechanisms employed by lineage‐determining TFs to direct gene programs controlling cell development and function.Support or Funding InformationNIH grant R01 DK098542This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Importance of Unique Secondary Structures in Genomic RNA1 3′ CITE in Blackcurrant Reversion Nepovirus Translation
Authors: Baquero Galvis Laura D. , Shields Elizabeth , Filbin‐Wong Megan E. 
Published: June 2021

Translation initiation and regulation commonly involve end‐to‐end interactions between proteins (initiation factors) bound to a 7‐methyl guanosine triphosphate cap at the 5′ end, and at a poly(A) tail, at the 3′ end of the messenger RNA. End‐to‐end communication is also prevalent in many RNA viruses, such as the Blackcurrant Reversion Nepovirus (BRV), which has two bipartite genomic RNAs (RNA1 and RNA2), both of which contain the 3′ poly(A) tail but lack the 5′ cap. The mechanism implemented by BRV during translation initiation is not clearly defined. In other cap‐less, positive‐sense RNA plant viruses, structured RNA in the 5′ and 3′ UTRs bind via proposed RNA‐RNA kissing‐loop interactions indicating that the recruitment of protein synthesis machinery is directly related to their primary, secondary and higher‐order structure. One form of non‐canonical (cap‐independent) translation involves structured RNA called the cap‐independent translation enhancers (CITEs). These components are proposed to bind to initiation factors as a 5′cap‐replacement and play an important role in the efficient translation of uncapped RNA genomes. Considering the end‐to‐end communication and recruitment of translation factors is dependent upon RNA structure, our main goal focuses on determining the secondary and possible tertiary structures of the 3′ CITE in the genomic RNAs found in the BRV, specifically RNA1. With this in mind, we have probed our purified RNA with a series of covalent nucleobase, sugar and backbone modifiers in addition to testing the function of a series of structural mutants via reporter assays. We found that the RNA1 CITE structure plays an important role in the process of viral protein synthesis by either facilitating end‐to‐end interaction with the 5′ UTR and/or directly recruiting important translation protein machinery. These findings shed light on structure‐based mechanisms used for end‐to‐end communication in non‐canonical mRNAs.Support or Funding InformationMSU Denver Colorado‐Wyoming Alliance for Minority Participation Undergraduate Research Grant, MSU Denver Dean's Grant, MSU Denver Provost GrantThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Yck3 Dependent Phosphorylation of Env7 and its Regulation During Cell Cycle in Saccharomyces cerevisiae
Authors: Manandhar Surya , Valencia Sara Patrice , Alvarado Carlos , Mansoora Ikha , Gharakhanian Editte 
Published: June 2021

Yeast vacuolar fusion and fission is a highly dynamic mechanism that responds to intracellular and extra‐cellular changes working to keep cells intact. This mechanism has been well conserved from yeast to humans. We discovered that Env7 is a conserved vacuolar membrane kinase that is involved in negative regulation of vacuolar fusion during budding and stress, similar to that of Yck3. We have shown that these two vacuolar kinases interact genetically and double delete shows the compromised phenotype [Manandhar, S; Gharakhanian, E., FEMS Yeast Res., 472–80, 2014] Here we also show that the double delete is defective in autophagy further supporting the possible interaction between these two genes. Based on gel shift assay, we have established that the phosphorylation of native Env7 is dependent on Yck3 and can be complemented by exogenously expressed Yck3 in an expression dependent manner. Here we also show that Yck3 directly interacts and phosphorylates Env7 at specific sites. By using site‐directed mutagenesis we have identified the Yck3 dependent phosphorylation sites of Env7. Our data strongly suggest that serine residues at positions 323 and 331 (S323 and S331) are the Yck3‐mediated phosphorylation sites of Env7 in consistent to the prediction by Bioinformatic analysis. Phosphorylation of Env7 is required not only for the stability of Env7 but also for regulating vacuolar morphology dynamics. Since we have shown that Env7 is essential for normal vacuoles dynamics during budding, studies in phosphorylation state and localization of Env7 during cell cycle arrest and progression using confocal microscopy are in progress. We are also currently working on exploring the possible downstream substrates that are likely involved in above mentioned cellular events.Support or Funding InformationThis project is funded by NIH‐SC1 GM112560‐01 grant to Dr. Editte GharakhanianThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Fyn‐dependent voltage‐gated potassium channel Kv1.3 modulates neuroinflammation and neurodegeneration in Parkinson's disease models
Authors: Sarkar Souvarish , Ngyen Hai Min , Malovic Emir , Langley Monica , Luo Jie , Singh Neeraj , Jin Huajun , Ananthram Vellareddy , Kanthasamy Arthi , Wulff Heike , Kanthasamy Anumantha 
Published: June 2021

A chronic neuroinflammatory event mediated by persistent microglia activation has been well recognized as a major pathophysiological contributor to the progression of neurodegenerative processes in Parkinson's disease (PD). Identification of key targets contributing to sustained microglia activation and the regulation of these targets could provide potential treatments to halt disease progression. In this study, we show that microglial Kv1.3, a voltage‐gated potassium channel, was highly upregulated in aggregated α‐synuclein (αSyn)‐stimulated primary microglia cultures, animal models of PD, as well as in human PD postmortem samples. Importantly, patch‐clamp electrophysiological studies confirm that the observed Kv1.3 upregulation translates to increased Kv1.3 channel activity. We further demonstrate that Fyn, a non‐receptor tyrosine kinase, modulated the transcriptional upregulation of microglial Kv1.3. Using multiple state‐of‐the‐art techniques, including DuoLink PLA technology, we show that Fyn directly binds to Kv1.3 and post‐translationally modified its channel activity. Furthermore, we demonstrate the functional relevance of Kv1.3 with respect to neuroinflammation by using Kv1.3 knockout (KO) microglia and the Kv1.3‐specific pharmacological inhibitor PAP‐1. Kv1.3 KO microglial cells treated with aggregated αSyn produced fewer pro‐inflammatory cytokines. PAP‐1 significantly attenuated aggregated αSyn‐induced inflammation in both a microglial cell line and primary microglia, thus highlighting Kv1.3's importance in inflammation. Oral administration of PAP‐1 significantly inhibited MPTP‐induced neurodegeneration and inflammation in vivo. PAP‐1 also significantly reversed behavioral deficits and dopamine loss in MitoPark mice, a progressive model of PD. Our results collectively show that the Fyn‐dependent Kv1.3 channel plays an important role in inflammation in PD and has potential therapeutic implications.Support or Funding InformationES026892, NS088206, NS100090, Llyod ChairThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ablation of Soluble Epoxide Hydrolase Expression Increases Beige Fat Over White Fat Through an Increase in Mitochondrial Integrity, Insulin Sensitivity and HO‐1‐PGC1α‐adiponectin in‐vitro and in‐vivo
Authors: Liu Lu , Vanella Luca , Bradbury J. Alyce , Zeldin Darryl C. , Abraham Nader G. 
Published: June 2021

Conversion of arachidonic acid epoxides to diols by soluble epoxide hydrolase (sEH) diminishes the beneficial cardiovascular properties of these epoxyeicosanoids. We therefore examined the possible biochemical consequences of inhibition of sEH and sEH deletion. We hypothesized that decreased sEH expression, would increase mitochondrial integrity, insulin sensitivity, EETs isomers and adipocyte function. To test this hypothesis, we examined the effect of sEH inhibitors on human MSC‐decreased adipocytes as well as sEH deletion in a mouse model treated with either HO‐1 lentiviral‐vector or cobalt protoporphyrin (CoPP) over an 8‐week period. Human MSc synthesized both EETs and EET metabolites (DHETs). MSc‐derived adipocyte treated with Auda, an inhibitor of sEH, exhibited a decreased number of large adipocytes, i.e. inflamed adipocyte but an increased number of healthy adipocytes that display increased levels of adiponectin, HO‐1 and PGC‐1a and decreased levels of TNFa. The addition of 8, 9, 14, and 15 EETs to adipocyte culture resulted in a reduction in large inflamed adipocytes, PPARg, FAS and lipid accumulation. In vivo studies demonstrated that sEH deletion resulted in a significant decrease in body weight and fasting glucose; p <0.05 and increased insulin sensitivity and serum adiponectin levels (p<0.03) and decreased visceral fat (p<0.05). Critically, sEH deletion was manifested by a significant increase in EETs isomers 5, 6; 8, 9; 11, 12; and 14, 15, EETs. Additionally, induction of HO‐1 potentiated the increases of EETs isomers, suggesting that increases antioxidant protected EETs from degradation by ROS. These novel observations highlighted the potential beneficial effects of EETs that resulted in increased mitochondrial integrity, HO‐1‐PGC1a and UCP1 levels. These results provide evidence that sEH deletion preserved vascular homeostasis, increased brown fat phenotype and that both EETs‐agonist and sEH inhibitors may have therapeutic potential in the treatment of metabolic syndrome and dyslipidemia.Support or Funding InformationNational Institutes of Health grant (HL34300 to NGA)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of sEH prevents high fructose‐induced impairement of myocardial oxygen comsumption in young rats
Authors: Froogh Ghezal , Kandhi Sharath , Duvvi Roopa , Weng Zan , Le Yicong , Alruwaili Norah , Ashe Johnathan , Sun Dong , Huang An 
Published: June 2021

High fructose diet increases tissue oxidative stress and has been shown to be detrimental to cardiac function. Epoxyeicosainoic acids (EETs) are metabolites of arachidonic acid, which possess cardioprotective properties and are metabolized by soluble epoxide hydrolase (sEH). Therefore, inhibition of sEH is proven to be beneficial to cardiovascular function. The specific impact of a high fructose diet and the role of EETs/sEH in the regulation of cardiac function in the adolescent population have not been studied. To test the hypothesis that sEH inhibition attenuates the adverse action of fructose consumption on the heart of young rats, 4 week old rats were fed a normal diet (as control; ND) or a high fructose diet (HFD) for 30 days, without or with concomitant administration of TPPU, a specific sEH inhibitor, by oral‐gavage (1mg/kg body weight: HFD‐TPPU).MethodsCardiac output was analyzed by echocardiography and blood pressure was recorded using tail‐cuff measurements. Myocardial oxygen consumption (MV̇o2) was measured in freshly isolated left ventricular tissue from all groups of rats under control conditions and following incubation of tissue with the nitric oxide (NO) synthase inhibitor Nω‐nitro‐l‐arginine methyl ester (L‐NAME; 10−4M), or NADPH oxidase inhibitor (Nox), apocynin (10−4M) for 1 hour. Cytosolic and mitochondrial superoxide levels were determined by HPLC‐fluorescence detection via incubation of myocardial tissues with dihydroethidium (DHE) or Mitosox. Nox 2 expression was measured via western blot analysis.ResultsFructose feeding significantly impaired MV̇o2 compared to hearts of ND rats, as indicated by the loss of NO‐mediated regulation of MV̇o2. Incubation of myocardial tissue isolated from the ND group with L‐NAME yielded the same pattern as fructose feeding, further confirming that the NO‐mediated response is compromised with high fructose consumption. The treatment of HFD rats with TPPU normalized their MV̇o2 to the level of ND rats. Furthermore, incubation of myocardial tissues isolated from HFD rats with apocynin elicited normalized MV̇o2, comparable to that observed in HFD‐TPPU rats. In addition, mitochondrial superoxide and Nox 2 expression were significantly increased in myocardial tissue of HFD rats compared to ND and HFD‐TPPU rats. Thus, we conclude that 1) high fructose‐induced increase in oxidative stress is responsible for impaired NO‐mediated regulation of MV̇o2. 2) Inhibition of sEH reversed the oxidative stress‐induced impairment of MV̇o2 via perhaps, increases in tissue levels of antioxidant EETs.Support or Funding InformationThis work was supported by HL070653 and HL129797This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.