bluuebunny/crossref_metadata_2025_split huggingface dataset:
Repeat Problems: Combinatorial Effect of C9orf72‐Derived Dipeptide Repeat Proteins
Authors: Darling April , Braydo Leonid , Blair Laura , Koren John , Uversky Vladimir 
Published: June 2021

The most common genetic cause of both Amyotrophic Lateral Sclerosis and Frontotemporal Dementia is a microsatellite expansion mutation in the 5′ UTR region of C9orf72. Expansion of the hexanucleotide repeat region of C9orf72 leads to loss of function, RNA foci, and five species of non‐AUG RAN translated dipeptide repeat proteins (GA, GP, GR, PA, & PR). It has been observed that the sense and antisense dipeptide repeat proteins can be translated simultaneously in one cell, however the interplay between the different species has not been heavily investigated. Here we determined that the antisense repeat protein PR is localized to the nucleus, causes cytotoxicity, disrupts nuclear transport, and triggers PERK phosphorylation and p62 puncta accumulation. However, when PR and its sense partner GA were co‐expressed, PR's localization and cytotoxic outputs were significantly altered. In addition, we showed that GA and PR interact through direct binding that leads to structural and morphological changes. We also demonstrate the ability of dipeptide repeat protein species to be released from one cell and taken up by another. Importantly, we demonstrate that co‐expression of distinct DPRs enhances the release of individual peptides. Thus, the combined expression of distinct C9orf72 derived dipeptide repeat species produces cellular outcomes and structural differences that are unique compared to the expression of a single species.Support or Funding InformationNot fundedThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dietary Glycine Supplementation Extends Lifespan of Genetically Heterogeneous Mice
Authors: Brind Joel , Miller Richard A. , Strong Randy , Harrison David E. , Macchiarini Francesca 
Published: June 2021

Preliminary experiments suggested lifespan extension by glycine supplementation (GlyS) without the growth inhibition seen with methionine restriction in Fisher 344 rats. This suggested that the benefit GlyS, rather than owing to increased methionine clearance (as was originally envisioned), was attributable to increased levels of glycine itself, acting via the glycine receptor. The glycine receptor is a glycine‐gated chloride channel that is widespread among cell types, including macrophages, and GlyS has been shown by others to exhibit anti‐inflammatory and anti‐cancer properties in several rodent models, and even to reverse Type 2 diabetes in a human trial. GlyS has also recently been found by others to restore the youthful mitochondrial phenotype in senescent cultured human fibroblasts, whose senescence was linked to epigenetic down‐regulation of the principal pathways of endogenous glycine synthesis. The present study set out to replicate the life extension previously observed in rats, in large, reproducible populations of UM‐HET‐3, 4‐way cross mice, in a Phase I Interventions Testing Program (ITP) trial of the National Institute on Aging (NIA). Beginning at 9 months of age, GlyS mice were fed a diet containing 8% glycine (dry weight, compared to the level of 2.3% in the control diet) simultaneously at 3 sites (The Jackson Labs, Bar Harbor, ME; University of Michigan, Ann Arbor, MI; U.T. Health San Antonio, TX), employing approximately 50 each of male and female GlyS and 100 each of male and female controls at each site. Data were compiled after 90% of control mice had died at all 3 sites. Pooling data from all 3 sites (shown in Figure below), median lifespan was increased among GlyS mice by 3.7% among females and 1.3% among males, and the 90th percentile lifespan was increased by 1.8% among females and 3.6% among males (p = 0.00004 by log‐rank test and p = 0.002 by Wang‐Allison test for pooled males and females at all 3 sites combined). The observed lifespan extension is statistically robust, in that significant log‐rank p‐values were seen for each sex (pooling across site) and for each site (pooling across sexes). The Stage I ITP protocol measures only lifespan, and it is anticipated that Stage II trials will provide the opportunity for biochemical and other cross‐sectional measures to elucidate the mechanism(s) involved in the life extension observed in our system.Support or Funding InformationInterventions Testing Program, National Institute on AgingThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regulatory mechanisms controlling the subcellular localization and activity of the RhoA GEF Net1 in breast cancer
Authors: Frost Jeffrey A. , Zuo Yan , Ulu Arzu 
Published: June 2021

Net1 is a RhoA/B‐specific guanine nucleotide exchange factor that is overexpressed in human breast cancer and is required for efficient cancer cell motility and invasion in vitro. Net1 knockout mice are viable but exhibit defects in tumor initiation and lung metastasis in a genetic model of breast cancer. Net1 is unusual among RhoGEFs in that it localizes to the nucleus in quiescent cells, but relocalizes to the cytoplasm in response to motility stimuli. However, regulatory mechanisms controlling its cytoplasmic accumulation are not well understood. In the present work we show that accumulation of the Net1A isoform in the cytosol requires CRM1‐dependent nuclear export and also requires phosphorylation by stress activated MAP kinases. Overexpression of constitutively active MKK7 or MKK3, which stimulate the Jnk and p38 MAPKs respectively, stimulates Net1A cytosolic localization in a ligand independent manner. Mutation of the main Jnk phosphorylation site, Ser52, to alanine prevents ligand stimulated cytosolic localization. Mechanistically, Net1A phosphorylation by Jnk prevents nuclear re‐import of cytosolic Net1A. Glutamic acid substitution of Ser52 promotes constitutive cytosolic localization and results in elevated RhoA activation and signaling. Expression of Net1A S52E promotes increased motility, extracellular matrix invasion, and invadopodia formation in breast cancer cells that are normally not invasive. Taken together, these data identify a novel regulatory mechanism controlling the subcellular localization and proinvasive functions of Net1A, and suggest novel therapeutic avenues to target Net1‐dependent cancers.Support or Funding InformationThis work was supported by NCI grant to CA172129 J.A.F. and a CPRIT fellowship RP160015 to A.U.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterization of Acid Phosphatase Adsorption to Montmorillonite
Authors: McTaggart William Stuart 
Published: June 2021

Phosphorus is important in agriculture because it limits the growth of plants and can influence the size of crop yields. The phosphorus cycle takes significant time in replenishing phosphorus lost to runoff and human activities — in the short‐term, phosphorus is non‐renewable. This project aims to better understand how naturally‐occurring enzymes influence the phosphorus cycle, particularly how these enzymes adsorb to soil. The project is taking advantage of microplate assays to analyze how acid phosphatase (AP) from wheat interacts with montmorillonite, a clay. Key experiments investigate the time it takes for the binding of AP to montmorillonite to equilibrate, the effect of AP concentration on adsorption and the total amount of binding. The data showed that binding came to equilibrium in approximately two hours, that greater binding occurs with more AP, and that the percentage of AP adsorbed increases significantly with the amount of montmorillonite. A fuller understanding of phosphorus cycling may allow for manipulation of the cycle to make phosphorus use more efficient.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Composition and viability of the microbial flora of the sigmoid colon obtained from embalmed human cadavers
Authors: Fracassi Rebeca Elizabeth , Holz Jonathan , Olivieri Marion Pace , Davie Jeremiah 
Published: June 2021

The gut microbiome contributes to proper digestion and nutrition. Abnormalities affecting the normal gut physiology can be responsible for disrupting the gut microbiome. Disruptions to the gut microbiome can be the cause of several pathologies. This research sought to determine the efficacy of utilizing cadavers for studying the gut microbiome. Previous studies utilizing DNA from fixed tissues for enzyme based assays, such as PCR were constrained by the chemicals in the fixative that researchers found it was difficult to eliminate these chemicals. The aim of this study was to determine if there were viable bacteria present in the sigmoid colon of embalmed cadavers. If present, these bacteria could be grown and expanded for further molecular based studies. The sigmoid colons of three male cadavers were isolated. The contents of the colon were assayed for the presence of culturable bacteria. Additionally, the sigmoid samples were sectioned for analysis of tissue morphology. These preliminary baseline histological studies suggest that the implemented embalming procedure of normal tissue architecture in the sigmoid colon resulted in elimination of all culturable bacteria. These results further suggest that, if viable bacteria are present, other methods are needed to fully access the presence of bacteria and microbiome in the sigmoid colon of embalmed cadavers.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 network approach to vertical transfer and articulation for student success in biology: A fourth workshop hosted by the Northwest Biosciences Consortium RCN‐UBE
Authors: Kruchten Anne , Baumgartner Erin , Beadles‐Bohling Amy , Brown Jeffrey , Duncan Jason , Kayes Lori , Kiser Stacey , Seidel Shannon , Shriner Walter , Stavrianeas Stasinos , Tillberg Chad 
Published: June 2021

The Northwest Bioscience Consortium (NWBC) is an NSF funded RCN‐UBE group working to align introductory biology curricula in the Pacific Northwest with the concepts and competencies of Vision and Change. During the last four years, the NWBC has hosted three workshops providing faculty development opportunities for colleagues across the region focused on classroom inclusivity, designing curricula for majors' introductory biology classes, and designing curricula for non‐majors introductory biology courses. We now report on our fourth winter workshop, entitled “A network approach to vertical transfer and articulation for student success in biology.” Held February 8–9, 2018, this workshop brought together faculty and administrative participants to collaborate on the NWBC model that all students are transfer students, whether they are transitioning from one institution to another or from introductory courses to advanced courses within the same institution. Day one of the workshop addressed issues affecting faculty, advisors, and administrators working with students in introductory biology courses, including state level initiatives mandating course topic coverage. The second day facilitated faculty groups working on common curricular outcomes to help students be successful in introductory biology courses and in vertical transfer to advanced biology courses. Here we present an overview of our workshop accomplishments and future plans for the NWBC and its RCN‐UBE funding.Support or Funding InformationThis work is funded by NSF RCN‐UBE 1248121.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mentorship for developing course‐based undergraduate research experiences (CUREs): The Council on Undergraduate Research (CUR) Mentorship for Integrating Research Into the Classroom (MIRIC) program
Authors: Wolyniak Michael J. , Resendes Karen K. 
Published: June 2021

The life science education community has responded to the recommendations of the American Association for the Advancement of Science (AAAS) Vision and Change document with several initiatives designed to improve the way in which undergraduates learn science. These initiatives have often taken the form of one‐time workshops that generate awareness of and interest in developing authentic research experiences for undergraduate STEM classrooms. However, they have been less successful with respect to generating the sustainable change necessary to bring real reform to undergraduate science education. To create sustainable change, long‐term faculty development initiatives focused on mentorship are needed so that instructors seasoned in developing and implementing course‐based undergraduate research experiences (CUREs) can convey their experiences to mentees interested in using these pedagogical techniques as the centerpiece of their own teaching. The Council on Undergraduate Research (CUR) Biology Division and American Society for Cell Biology (ASCB) Education Committee are creating the Mentorship for Integrating Research Into the Classroom (MIRIC) program to provide a means for members with an interest in developing improved and sustainable active learning techniques to gain experience in this style of teaching through close, long‐term interaction with a veteran teaching mentor.Developed from the former ASCB Mentorship in Active Learning and Teaching (MALT) program, MIRIC focuses on the development of instructors who wish to develop a dynamic CURE. Current and future life science instructors pair themselves up with seasoned veterans of CURE development and work with them and their students over the course of a semester or longer to develop a CURE that will allow the mentee to bring authentic research into his or her classes.In our pilot studies, we collected qualitative and quantitative data based on participant interviews and coding videos of student and instructor actions during classroom activity (Smith et al., 2013), respectively, that suggest that MIRIC/MALT mentorships have made positive gains in promoting sustainable active learning techniques among participants. Going forward, we wish to use instruments like the Laboratory Course Assessment Survey (Corwin et al., 2015) and Experimental Design Ability Test (Sirum and Humburg 2011) to assess the effectiveness of the MIRIC laboratory intervention.MIRIC will undergo its full launch in 2018. At present, please consult http://www.ascb.org/mentoring‐in‐active‐learning‐and‐teaching‐malt/ for more information on becoming involved.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Graduate student professional development and a CURE‐style course and peer‐reviewed student publications
Authors: Baumgartner Jackson , Lee Jesi , Kuhn Misty L. 
Published: June 2021

Implementing Classroom Undergraduate Research‐based Experiences (CUREs) into an institution's curriculum has shown measurable benefits for students in the chemical sciences. However, several areas in biochemistry, such as protein structure, have shown resistance to a similar scale of implementation. Here, we describe a graduate level CURE‐style pilot program on protein structure/function that was held at San Francisco State University. In this class, first and second year Master's students investigated four individual crystal structures of proteins of the dTDP‐L‐rhamnose biosynthetic pathway. The students examined the biochemical and structural background literature of their assigned proteins, visualized and analyzed protein 3D structures using in silico methods, and wrote a manuscript for submission for publication. A weekly “show‐and‐tell” was held during each session where the students shared their findings and expertise with classmates. Students worked individually and in groups, which mimicked the free exchange of ideas and feedback typical of a scientific community's communication process. We chose structures of each protein that belonged to a single metabolic pathway to ensure multiple types of protein structures were observed, analyzed, and discussed during the course. Additionally, it provided a cohesive biological context for discussion and understanding of how proteins within a metabolic pathway coordinate to complete a cellular process. A manuscript‐style final paper and poster presentation at the University's annual research poster showcase were the primary classroom assessments for the course. One caveat of this approach regarding publication of results is that the instructor must be able to collaborate with the original structure depositor. Overall, this CURE‐like graduate course provided Master's students with a unique opportunity to experience the authentic preparation of a research publication, which is indispensable for preparing them for their future scientific careers. Three of the four manuscripts that were submitted for publication are currently in press.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mutation in MON2, Which Encodes a Protein Implicated in Vesicular Transport, Affects Response to Exogenous Fatty Acids in Yeast
Authors: Draper Riley , Gasparotto Anthony , Soni Mona , Gasper Brittany , Stukey Joseph , McDonough Virginia 
Published: June 2021

Metabolism of exogenous fatty acids requires cells to take up, traffic, and molecularly respond to fed fatty acids. Our lab has determined that a mutation in mon2 results in defective growth on the fatty acid 10‐undecenoic acid. Like wild type cells, mon2Δ mutants are capable of normal response to the feeding of long chain unsaturated fatty acids (UFAs) by reducing transcription of OLE1, (which encodes the Δ9 desaturase), and by reducing the accumulation of endogenously produced UFAs. However, wild type cells when fed 10‐undecenoic acid will continue to express OLE1 and increase their content of endogenously produced UFAs. In contrast, mon2Δ mutants do not differentially respond to the two fatty acids. Staining with a BODIPY‐labeled fluorescent fatty acid analog indicates that there is a difference in localization/amount of the supplied fatty acid in mon2Δ cells compared to wild type cells. Thus, it appears that the mon2 mutation decreases the cell's ability to differentiate between fatty acids, perhaps due to mislocalization of the fed fatty acid or regulatory proteins.Support or Funding InformationThis work was supported by a Jacob Nyenhuis Faculty Award (VM), NSF‐REU (AG, MS, BG) and a Summer Scholar award from the Dean for Natural and Applied Science (RD).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Human Follicle Stimulating Hormone Receptor Lipid Raft Residency is Hormone and Caveolin Dependent
Authors: Madden Hannah E. , Stewart Angelina , Cohen Brian D. 
Published: June 2021

Human follicle stimulating hormone (hFSH) is a gonadotropin responsible for the maturation of ovarian follicles in women and production of sperm in men. The receptor (hFSHR) is a G protein‐coupled receptor (GPCR); a class of receptors found embedded in the cell membrane to receive the hormone resulting in initiation of a complex downstream signaling cascade. Through previous research in the Cohen lab, hFSHR has been shown to be located within compartments of the cell membrane known as lipid rafts. Lipid rafts are microdomains of the membrane containing a higher concentration of cholesterol, glycosphingolipids, and a protein known as caveolin. Current research is focused on evaluating the mechanisms and effects of hFSHR lipid raft residency. Using discontinuous sucrose gradients we have shown that hFSH stimulation of an HEK293 cell line stably transfected with hFSHR cDNA results in alteration of hFSHR lipid raft residency in a time dependent manner.To better understand the mechanism and effect of hFSHR lipid raft residency the same cells were treated with a peptide mimetic of transmembrane domain 4 of hFSHR. This peptide includes a sequence consistent with a motif shown in other proteins to interact with caveolin (ϕXϕXXXXϕXXϕ where ϕ is an aromatic amino acid). The putative caveolin interaction motif of hFSHR was synthesized along with the HIV‐TAT leader sequence to allow passage through the membrane. A second peptide where the aromatic amino acids were replaced with leucine was also used. Cells treated with the wild type peptide showed elevated basal signaling in the absence of hFSH, an effect not observed with the mutant peptide.Together, these results suggest that hFSHR residency in lipid rafts is dependent on hormone treatment and interaction with caveolin. Through studying hFSHR lipid raft residency and signaling we hope to identify ways to control the receptor for improved methods for contraception and treatment of infertility.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Isolation of Phagosomes Containing Bovine Photoreceptor Outer Segments from Retinal Pigment Epithelial Cells by Magnetic Selection
Authors: Dorvilier Rachelle , Shelby Shameka 
Published: June 2021

Daily phagocytosis of the distal tips of photoreceptor cells by the retinal pigment epithelium (RPE) is known to be an essential mechanism in the maintenance of the retina. The outermost portions of the photoreceptor cells, termed outer segments, are ingested and subsequently processed by the RPE. Previous studies involving the isolation of phagosomes have utilized latex beads to induce phagocytosis in macrophages, which lacks specificity due to the large foreign body being ingested. Therefore, the purpose of this study was to develop a standard protocol for the isolation of outer segment enriched phagosomes from RPE cells. To generate phagosomes with the selected cargo, outer segments isolated from bovine retinas were covalently labelled with magnetite particles and then fed to RPE‐J cells. After feeding, the cells were washed and gently lysed with a Dounce homogenizer. The phagosomes containing the magnetite bound outer segments were collected via magnetic rack isolation and stored in a homogenization solution. Immunoblots using antibodies against Mertk, N and C‐terminal rhodopsin, and cathepsin D confirmed the presence of outer segment‐containing phagosomes of varying maturity in the isolated fraction. The protocol we have developed has successfully isolated outer segment enriched phagosomes from RPE‐J cells by magnetic selection. This study differentiates itself from other studies that employ latex beads to isolate phagosomes through density gradient centrifugation. The isolation of these phagosomes will potentially play an important role in investigating protein interactions that impact RPE membrane trafficking and maturation.Support or Funding InformationFlorida Southern College Faculty‐Student Collaborative Research GrantThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.