bluuebunny/crossref_metadata_2025_split huggingface dataset:
Novel Regulation of Lipid Metabolism by a Phosphatidylinositol Transfer Protein and a Phosphatidylinositol 4‐Kinase
Authors: Wang Yaxi , Yuan Peihua , Rodriguez Martin , Tripathi Ashutosh , Lönnfors Max , Schuldiner Maya , Moye‐Rowley W. Scott , Bankaitis Vytas 
Published: June 2021

Membrane contact sites (MCSs) are regions in cells where membranes of two organelles are in close apposition. MCSs are proposed to facilitate non‐vesicular trafficking of lipids. Lipid transfer proteins (LTPs) are one of the most commonly‐found components in the MCS system and are speculated to mediate lipid transfer between membranes. However, both the biological functions of MCSs and the functional role of LTP in MCSs remain largely elusive. Herein, we examine in detail a previously‐proposed MCSs model, the ER‐Golgi/endosome MCS facilitating the phosphatidylserine (PtdSer) decarboxylation 2 (Psd2) pathway, which converts PtdSer into phosphatidylethanolamine (PtdEtn) in Saccharomyces cerevisiae. Particularly, we investigate the biological role of a yeast LTP, one of the yeast Phosphatidylinositol (PtdIns)‐Transfer Proteins (PITPs), Sfh4, and a PtdIns 4‐OH kinase Stt4, in this Psd2‐MCS system.We found that Psd2, Sfh4, and Stt4 are the only essential components facilitating Psd2‐dependent PtdEtn synthesis while other previously proposed elements constituting the Psd2‐MCS (Scs2, Scs22, and Pbi1) are dispensable. Surprisingly, neither the PtdIns‐transfer activity of Sfh4 nor its capacity to activate Stt4 is required to stimulate the Psd2 pathway. Instead, Sfh4 activates the Psd2 pathway via an Sfh4‐Psd2 physical interaction involving the F175 residue on the surface of Sfh4. Stt4 also displays physical interaction with Psd2 in an Sfh4‐independent manner. Instead of directly activating Psd2, Stt4 regulates the substrate pool accessible to Psd2, thereby indirectly controlling the Psd2‐mediated synthesis of PtdEtn. These results demonstrate that the ER‐endosomal MCS model is an inaccurate description of the Psd2 system in yeast, and provide an outstanding example where PITP biological function is uncoupled from its ‘canonical’ activity as a PtdIns transfer protein. Additionally, our findings reveal a novel mechanism in which a PITP and a PtdIns 4‐OH kinase regulate cellular phospholipid homeostasis.Support or Funding InformationThis work was supported by grants from the National Institutes of Health (GM44530) and the Robert A. Welch Foundation (BE‐0117) to VAB. The authors declare no financial conflicts.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sulforaphane Induces Colorectal Cancer Cell Proliferation through Nrf2 Activation in a p53‐Dependent Manner
Authors: Oh Jisun , Kim Jong‐Sang 
Published: June 2021

Sulforaphane is a well‐known phytochemical that stimulates nuclear factor erythroid 2‐related factor 2 (Nrf2)‐mediated expression of antioxidant and detoxifying enzymes. In this study, we found that sulforaphane promotes cell proliferation in HCT116 human colon cancer cells harboring a normal p53 gene in a dose‐dependent but biphasic manner. Since p53 has been reported to contribute to cell survival by regulating various metabolic pathways to adapt to mild stress, we further examined cellular responses in both p53‐wild‐type (WT) and p53‐knockout (KO) HCT116 cells exposed to sulforaphane in vitro and in vivo. Results demonstrated that sulforaphane treatment (i) activated Nrf2‐mediated antioxidant enzymes in both p53‐WT and p53‐KO cells, (ii) decreased apoptotic protein expression in WT cells but increased in KO cells in a dose‐dependent manner, (iii) marginally changed mitochondrial DNA abundance in WT cells but decreased in KO cells, and (iv) increased the expression of a mitochondrial biogenesis marker PGC1α in WT cells but decreased in KO cells. Moreover, a low dose of sulforaphane promoted tumor growth, upregulated the Nrf2 signaling pathway, and decreased apoptotic cell death in p53‐WT HCT116 xenografts compared to that in p53‐KO HCT116 xenografts in BALB/c nude mice. These findings suggest that sulforaphane can influence colon cancer cell proliferation and mitochondrial function through a crosstalk between the Nrf2 signaling pathway and p53 axis.Support or Funding InformationThis work was supported by the National Research Foundation of Korea (NRF) grant funded by the Ministry of Science and ICT (MSIT), Republic of Korea (Grant No. 2017R1A2B4005087).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Delineating the Link Between Dilated Cardiomyopathy and Arrhythmogenic Symptoms
Authors: Muser Tyler , Feyen Dries , Bruyneel Arne , Mercola Mark 
Published: June 2021

Dilated cardiomyopathy (DCM) is a common cause of heart failure and frequently involves life‐threatening ventricular arrhythmia and sudden cardiac death. In addition, DCM patients often show a heightened susceptibility to drug‐induced arrhythmia. The mechanisms that link DCM to ventricular arrhythmia are not well understood but are thought to involve structural alterations in the heart wall and in the ion channel makeup of individual cardiomyocytes. About 50 genetic mutations are known to cause congenital forms of DCM. Induced pluripotent stem cell‐derived cardiomyocytes (iPSC‐CMs) generated from patient tissue recapitulate many pathophysiological phenotypes characteristic of congenital heart disease and offer an unprecedented tool to study disease mechanisms. To model DCM, we used iPSC‐CMs harboring a DCM‐causing mutation in phospholamban (PLN), a protein involved in calcium regulation. Using the PLN mutant cells, we tested whether the disease would increase arrhythmic susceptibility to drugs that target certain ion channels. Initial experiments from my internship found that both Sodium and human Ether‐à‐go‐go Related channel agonists elicited an exacerbated proarrhythmic response from mutant cells. Action potential durations were disproportionately increased in the DCM‐afflicted mutants. If these data are sustained throughout the experiment, they support the model that the PLN mutant DCM involves a channelopathy that increases proarrhythmic sensitivity to drugs.Support or Funding InformationTM acknowledges support from the American Heart Association and the Stanford Cardiovascular Institute.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A comparative analysis between PrPc and PrPsc to determine the structure‐function relationship of protein misfolding in Scrapie prion disease
Authors: Peterson Alison , Borisova Catherine , Bunday Priscilla , Gong Meili , Merlin Daniela Hernandez , Kachkovsky Nicholas , Quist Eli , Yeoh Ming Ying 
Published: June 2021

The Minnetonka MSOE Center for BioMolecular Modeling SMART Team used 3D modeling and printing technology to examine structure‐function relationships of PrPsc. Scrapies is caused by prions, misfolded proteins in animals and humans, resulting in the development of fatal neurodegenerative encephalitis. Scrapies was one of the first prion diseases to be isolated and studied, and was found to be caused by the misfolding of the PrP protein into the PrPsc prion. We will do this by creating, annotating, and comparing two models, one of the original PrP protein, and one of the PrPsc prion. Scrapies disease is caused when the alpha‐helix rich prion protein (PrP) is converted into into a beta‐structure‐rich insoluble conformer (PrPsc) and is thought to be highly infectious. These structural changes in PrPsc contribute to its build up in the brain of sheep and goats PrP, leading to a deadly swelling of the brain. The buildup is caused by the inability of proteinase K (PK) to fully degrade PrPsc. PK has the ability to fully degrade PrP, but, interacting with PrPsc, can only create N terminally truncated PrPsc, resulting in a PK resistant core. There is currently no cure for Scrapies; however, further research is being done to discover how to reverse the functional effects of prion diseases. Our goal is to present which structural changes occur in the original protein PrP that cause the defective protein PrPsc, as well as determine how this misfolding occurs and what effect it has on the protein's function.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Elucidating the Role of an Atypical Anti‐Termination Two‐Component System in Mycobacterial Gene Regulation and Pathogenesis
Authors: Davenport Haley , Rohde Kyle 
Published: June 2021

M. tuberculosis, the etiologic agent of tuberculosis, caused 1.3 million deaths and infected 10 million people globally in 2017. During infection, many bacteria respond to their environments using two‐component systems (TCS) consisting of a membrane‐bound histidine kinase (HK) and a response regulator (RR) of transcriptional initiation. Rv3220c (HK) and Rv1626 (RR) are an atypical TCS with a cytosolic HK and an RR acting as a transcriptional anti‐terminator via its ANTAR domain. This domain causes formation of an anti‐terminator structure to allow transcription to proceed past its normal termination point. Rv1626 was previously identified by a transposon screen as involved in cholesterol metabolism, but the role of this TCS in pathogenesis is unknown.To determine the function of this TCS, knock‐out (KO) constructs were created using PCR, TOPO cloning, and recombineering. Due to the safety, faster growth, and genetic similarity of M. smegmatis to M. tuberculosis, we targeted the orthologs of the HK and RR in this surrogate organism, MSMEG_1918 and MSMEG_3246 respectively, for inactivation. After generating KO mutants, a phenotypic study was done to determine if cholesterol metabolism was inhibited by the gene deletion. Secondly, mycobacterial protein fragment complementation was used to confirm protein‐protein interactions between the putative TCS proteins. Findings from M. smegmatis will be validated in future work by conducting similar studies in pathogenic mycobacteria.Support or Funding InformationThe project was supported by startup funds to Dr. Kyle Rohde from University of Central Florida College of Medicine.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Prognostic Value of Ephrin Type‐A2 Receptor (EPHA2) and Ki67 in Renal Cell Carcinoma Patients: An Immunohistochemical and Bioinformatical Approach
Authors: Talaat Iman Mamdouh , Okap Israa Sobhy , El Sheikh Samar Mohamed , Youssif Tamer Mohammed Abou , Hachim Ibrahim Yaseen , Hachim Mahmood Yaseen 
Published: June 2021

BackgroundThe incidence of renal cell carcinoma (RCC), the most common malignant renal epithelial tumour, is increasing worldwide. Patients usually present with advanced disease and mostly have unpredicted clinical behaviour. A variety of prognostic factors have been proposed as useful parameters, however many of them showed limited clinical value. Thus, the discovery of novel prognostic markers, which might help in predicting patients' outcome and additional new targets to treat this disease are still in great demand. EphA2, a receptor tyrosine kinase, was found to be overexpressed in several malignancies and its expression in different tumours was found to be associated with poor prognostic features. In addition, a group of emerging strategies was recently introduced to target this receptor and found to show anti‐tumorigenic activity in various pre‐clinical cancer models.AimTo investigate the prognostic value of EphA2 expression in RCC patients and its association with other clinicopathological parameters as well as Ki67 expression, which is a well‐known proliferative and prognostic marker.Materials and MethodsThe expression of EphA2 and Ki67 was investigated immunohistochemically, and the results were correlated with the different clinicopathological parameters in 50 primary tumour blocks obtained from RCC cases surgically managed in the Urology Department, Alexandria University Main Hospital, during the period (2012–2015). In addition, the association between the EphA2 mRNA expression and the tumour stage as well as the patient outcome was also evaluated using two large publicly available databases.ResultsOur results showed a significant association between EphA2 expression and the tumour size, the nuclear grade, the tumour stage, the patient's outcome and the Ki‐67 expression (P < 0.05 for all). The same trend was also observed with EphA2 mRNA expression using larger patients' cohorts in two publicly available databases. Notably, EphA2 protein expression showed higher levels of co‐expression with the proliferative marker Ki67.ConclusionWe concluded that higher expression of EphA2 and Ki67 in tumour tissues predicts a locally aggressive behaviour and poor outcome of RCC patients. Moreover, our results give a rationale for the potential benefits of using novel therapeutic strategies with the aim of targeting EphA2 receptor in RCC that might help in improving their outcome.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evaluation of Dry Eye Syndrome Induced by Urban Particulate Matter Using Aerosol Exposure System
Authors: Lee Tae Gu , Hyun Soo‐Wang , Park Bongkyun , Jo Kyuhyung , Lee Ik Soo , Kim Jin Sook , Kim Chan‐Sik 
Published: June 2021

Dry eye syndrome (DES) is caused by multi‐factors and it is featured as tear deficiency and eye irritation. Air pollutions, represented as particulate matter (PM), are emerging threat of human health which induce various diseases including DES. However, the pathogenic mechanisms of DES via PM exposure are not completely identified. Though, several in vivo models of PM induced DES have been suggested, these were not properly reflect the phenomena of human who exposed to PM. In this study, we utilized inhalation device to mimic the atmospheric environment of PM exposure. Rats were exposed to urban PM (UPM) for 5 hours a day during 5 days. After exposure, tear volume was reduced in UPM exposure group. In addition, corneal irregularity and corneal epithelial cell apoptosis were significantly increased by UPM exposure. UPM also affected the tear film disruption via mucin‐4 reduction. Our purpose in this study was to demonstrate the application of new in vivo exposure model for DES induced by PM. We suggest that aerosol exposure system of PM can be used for risk assessment of DES.Support or Funding InformationThis study was carried out with the support of ‘R&D Program for Forest Science Technology (Project No. 2017035A00‐1919‐BA01)’ provided by Korea Forest Service (Korea Forestry Promotion Institute). This work was also supported by Korea Institute of Oriental Medicine (Project No. KSN1812080).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Optimizing Student Use of Quiz Questions as a Learning Tool
Authors: Peffley Dennis , Di Rito Meghan , Bickett Skye , Lee Jiehyun M 
Published: June 2021

OBJECTIVEStudents often utilize practice questions to improve understanding and retention of session content and to prepare for exams. However, a major challenge is aligning questions with session learning objectives. The goal of this study is to change student behavior to use questions as a learning tool, create intuitive question searching at the correct level, and align commercial and professor‐written question banks.METHODA developmental model was created combining faculty‐generated questions with commercial question bank questions. Three surveys were sent out to assess perceptions of question bank utilization. The first two surveys were sent out simultaneously, one to faculty and one to students, to assess how practice questions were used for study purposes. Following the developmental model, a pilot study was created using two practice quizzes with faculty‐generated practice questions for a pharmacy anatomy course. After the completion of the pharmacy anatomy course, the third survey was sent out to pharmacy students to assess their perception of the utility of practice quizzes and to assess whether student utilization of quizzes had changed.RESULTSTwenty‐three percent of faculty responded to the survey, indicating that most faculty create their own questions, but do not direct students to use commercial question banks. Seventy‐two students (a 12.04% response rate) responded to the first survey, indicating that 53% of students use faculty provided questions while 40% use commercial question banks and 34% use both. Most students reported using question banks to study the week before a test, three days before, or the day before the test. The first survey indicates that roughly 15% and 5.6% of students quiz themselves the same week of the lecture and the day of the lecture, respectively. In comparison, the post‐survey after the pilot study with a 15% response rate demonstrates that students began studying earlier using quiz questions. The post‐survey indicates 28.6% and 7.1% of students in the pilot study test themselves the week of the lecture and the day of the lecture, respectively. The percentage of students who quizzed themselves a week before the test also jumped from 44.4% to 57.1%. Additionally, in the original survey, only 37.5% of students said that they always incorporate practice questions as part of their studying. In the follow up survey, this jumped to 50%.CONCLUSIONThe initial survey showed students inhibit their study strategies by not quizzing themselves early enough. However, the curated set of progressive questions helped students incorporate question banks into their studying. Additionally, they used the question banks to start studying more effectively by quizzing themselves earlier.Support or Funding InformationPhiladelphia College of Osteopathic Medicine GeorgiaThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
EXAMINING THE PRE‐POST MATRICULATION ACADEMIC METRICS AS CORRELATES OF OSTEOPATHIC MEDICAL STUDENT CLINICAL PERFORMANCE
Authors: Eldeeb Khalil , Dogbey Godwin 
Published: June 2021

PURPOSEThe clinical years of medical school are critical in the training of medical students. It is a period that informs and shapes their professional decisions. Results from studies have been perverse about students' academic metrics that predict their clinical success. This study seeks to examine the academic performance metrics that are correlates of osteopathic medical students' clinical performance.METHODSThis is an IRB approved study involving a retrospective review of admission and academic performance records of the first‐two graduated student cohorts from the Campbell University Jerry M. Wallace School of Osteopathic Medicine in North Carolina. For categorical variables, summary descriptive measures—frequencies/percentages will be generated while means, standard deviations/ranges computed for continuous variables. Correlational analyses will be conducted between the academic and clinical performance variables. Secondary exploratory analyses will be conducted inferentially as necessary. All statistical inferences will be considered significant whenever, P ≤0.05.RESULTS (Preliminary)Entering science, non‐science and total GPA are positively correlated with third and fourth‐year medical school GPAs. While entering science and total GPA are positively correlated with COMLEX‐USA Level 2‐Cognitive Evaluation (CE) total score they are not positively correlated with any of the Clinical Sciences Disciplines scores or COMLEX‐USA Level 3 total score or any of the Clinical Sciences Disciplines scores. Entering non‐science GPA is positively correlated with COMLEX‐USA Level 3 ‐ Family Medicine only.While the Physical component of the MCAT is not correlated with enrolled third and fourth‐year medical school GPAs or board exams level 2 or 3, the verbal component is positively correlated with fourth‐year medical school GPAs only. The biology component and the Total MCAT score of the MCAT is positively correlated with both COMLEX‐USA Level 2‐Cognitive Evaluation (CE) total score and COMLEX‐USA Level 3 total score. The biology component also positively correlate with level 2 family and internal medicine and level 3 emergency and internal medicine scores. The third and fourth‐year medical school GPAs are positively correlated with both COMLEX‐USA Level 2‐Cognitive Evaluation (CE) total score and COMLEX‐USA Level 3 total score and all Clinical Sciences Disciplines scores.CONCLUSIONPreadmission academic variables are associated with matriculation academic and second and third‐level national board examinations performance of osteopathic medical students. Admission processes should consider the importance of these metrics in student recruitment to maximize their clinical success post‐matriculation.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interaction between Calmodulin and Lysosomal Vesicle‐Associated Membrane Protein 8
Authors: Yao Annie , Bloomfield Christina , Tran Quang‐Kim 
Published: June 2021

Fusion of intracellular membranes is an important step in key processes such as exocytosis. This process is directly controlled by multiple vesicle‐associated membrane proteins (VAMPs). However, its regulatory inputs remain incompletely understood. Calmodulin is the key transducer of intracellular calcium signals that has been shown to control neurotransmitter release via its interaction with VAMP2. VAMP8 is a distinct protein located on the lysosomal membrane and is important for fusions of intracellular membranes. We hypothesized that calmodulin directly interacts with VAMP8, thereby controlling lysosomal fusion. In this study, we identified the calmodulin‐binding domain on VAMP8 and characterized the biochemical properties of the interaction.Using a new biosensor method to screen for calmodulin‐binding sites, we identified a 20‐a.a. domain on VAMP8 as its calmodulin‐binding domain; truncation of three C‐terminal residues on this sequence significantly reduced binding affinity. To determine the concentrations of calcium at which this interaction can occur, we simultaneously monitored the responses of VAMP8 biosensors and a spectrally suitable calcium indicator in the presence of saturating calmodulin concentration and incremental addition of calcium. Control experiments demonstrated absolute absence of bleed‐over in this system. These determinations revealed that calmodulin interaction with VAMP8 is calcium‐dependent and can occur across the physiological range of cytoplasmic calcium concentration.These data indicate novel roles for calcium and calmodulin in regulating VAMP8 function. The results also suggest potential roles in autophagy for factors that control calcium signals, calmodulin expression and its dynamic target interactions.Support or Funding InformationNational Institutes of Health Grant HL112184 and Des Moines University Grant IOER‐090717This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.