bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Locomotor Activity Study of the Effects of L‐Serine to Neurodegeneration Impacts of Beta‐Methylamino‐L‐alanine in Drosophila melanogaster
Authors: Nguyen Quang Thanh , Rivera Lemuel Vince , Goto Joy June 
Published: April 2020

Amyotrophic Lateral Sclerosis‐Parkinsonism Dementia Complex (ALS‐PDC) is a neurodegenerative disease that described by loss of motor function, tremors, and dementia due to neurodegeneration. The disease is thought to be caused by Beta‐Methylamino‐L‐Alanine (BMAA), a neurotoxin from blue‐green algae. Since it has a structure that is similar to glutamate, BMAA act as an agonist to the glutamate receptor N‐methyl‐D‐aspartate which lead to excitotoxicity and degenerate neurons. L‐serine is an essential amino acid that has shown to interfere with the effect of BMAA in vivo studies. The purpose of this research is to further test the novel impact of L‐serine to BMAA‐induced fly in long‐term study. The interaction of BMAA and L‐serine was analyzed through circadian rhythm activity and viability of wild‐type Drosophila melanogaster in ALS‐PDC model. We used two Drosophila Activity Monitor (DAM) systems to measure activity of 64 fruit flies (Gender and Age‐Match). The experimental groups were fed with L‐serine, BMAA, BMAA+L‐serine and the control group was fed with standard fly agar. Fly activity was recorded by the number of fly pass the IR beam in the monitor for 10‐day (12L/12D at 22°C). After 10 days, circadian rhythm data showed BMAA flies increased activity over the first few days, and decreased gradually to the end of the period. BMAA‐induced flies (n = 12) does not show pattern of regular activity in L/D as compared to Control (n = 15) and L‐serine (n = 16). The co‐fed BMAA+L‐serine (n = 7) showed a similar pattern as the Control. For daytime vs nighttime activity (mean ± s.e.m), Control (n = 15) shown regular activity (day: 137 ± 13%, night: 21 ± 5%) as compared to BMAA (n = 12) that shown high activity in dark cycle (day: 162 ± 10%, night: 155 ± 21%). Moreover, the cofed L‐serine + BMAA (n = 7) exhibited normal daytime activity but not nighttime activity (day: 138 ± 14%, night: 167 ± 31%). The data from DAM shows the abnormal activity of flies in long term when inducing with BMAA such as abrupt sleep due to high activity during dark phase. L‐serine shows to reduce down the high activity when cofed with BMAA. This supports our hypothesis of L‐serine ameliorates the effects of BMAA in fly. This preliminary data is a starting point for further research in drug discovery for ALS‐PDC.Support or Funding InformationAcknowledgementsThis work was funded by the Fresno State Undergraduate Research Grant Award (2019–2020), and Howard P. Kubo, M.D. Award for Outstanding Achievement in Biomedical Science.
Identification Of An Actin Fold Containing Metabolic Enzyme That Undergoes Regulated Polymerization
Authors: Wilhelm Jim , Carver Michael , Kyriakakis Phillip , Monfort Elena , Barry Rachael , Leschziner Andres 
Published: April 2020

Cytoskeletal actins are thought to have evolved from actin fold enzymes, such as hexokinase or hsp70, that acquired the ability to polymerize while losing all other enzymatic functions apart from ATP hydrolysis. However, the discovery of metabolic enzymes, such as CTP synthetase, whose enzyme activity is regulated by self‐assembly into filaments has suggested another route to evolving cytoskeletal polymers: a filament forming metabolic enzyme that lost its role in metabolism and acquired novel cellular functions. In order to investigate this possibility, we surveyed all of the actin fold sugar kinases in yeast for their ability to form filaments. This screen identified one metabolic actin, the yeast glucokinase, Glk1p, that was capable of forming filaments in vivo. We have reconstituted Glk1p polymerization in vitro and have found that either ATP/Glucose or glucose‐6‐phosphate is sufficient to trigger filament formation. Consistent with this, we have also found that increasing glucose‐6‐phosphate levels in vivo triggers Glk1p polymerization. Furthermore, the addition of glucose and nonhydrolyzable ATP analogs to Glk1p also causes filaments to form. Thus, ATP hydrolysis is not required for Glk1p polymerization. Interestingly, the addition of ADP causes rapid disassembly of glucose‐6‐phosphate/Glk1p filaments suggesting that the stability of the filament is sensitive to the hydrolysis of ATP. While this is reminiscent of the dynamic behavior of cytoskeletal actins, our studies of Glk1p argue that it forms filaments that are structurally distinct from cytoskeletal actin filaments. Since the early steps of glycolysis consume ATP before generating ATP, we propose that Glk1p polymerization acts as a brake on glycolysis to prevent catastrophic depletion of ATP under certain metabolic conditions. We are currently using our structural and biochemical insights into Glk1p polymerization to test this possibility. Because Glk1p is the first filament forming actin that has more enzymatic functions than simple ATP hydrolysis, our results provide novel insights into both the evolution of the actin cytoskeleton and how the plasticity of the actin fold can support both polymerization and essential enzymatic activities.Support or Funding InformationThis work was supported by a HHMI Collaborative Innovation Award
The Lipid Anticancer Drug Edelfosine Affects Gene Expression Regulation
Authors: Ponce Maria Laura Sosa , Cobb Jennifer A , Zaremberg Vanina 
Published: April 2020

Sirtuins are highly conserved regulators of gene expression. A genetic screen performed by our group using budding yeast found that sensitivity to the anticancer lipid drug edelfosine is alleviated in cells lacking sirtuins (SIR complex in yeast). The mechanism by which sirtuins mediate sensitivity to edelfosine, which localizes to the nuclear envelope (NE), is unclear. Here, we investigated the effect of edelfosine on the nuclear membrane and regulation of the nucleus. Using fluorescence microscopy, we observed NE deformation and changes in clustering of the telomere‐binding proteins Rap1 and Sir4. Transcriptomic analysis of cells treated with edelfosine showed major downregulation of genes known to be bound by Rap1, with enrichment of ribosomal protein genes. The overall effect of edelfosine on gene expression was reduced in sir4Δ mutant cells. We also found that constitutive deacetylation of histone H4 Lys16, a target of the SIR complex, caused hypersensitivity to edelfosine. We speculate that edelfosine disrupts genomic regulation via a SIR‐mediated mechanism. We show here that lipid anticancer drugs can affect DNA regulation by disrupting the NE, which has interesting implications for potential combination therapies.Support or Funding InformationThis work has been financially supported by the Natural Sciences and Engineering Research Council of Canada to VZ and JAC. Also supported by a QEII scholarship to MLSP.
Evaluating the Metabolite Profile of HER2‐positive Breast Cancer Cells
Authors: Abuelreich Sarah , Leon Nicole , Medina Daisy , Sahni Malika , Krishnan Krish , Bush Jason 
Published: April 2020

Human epidermal growth factor receptor 2 (HER2)‐positive breast cancer represents about 20% of breast cancer cases and although treatments are available, some patients develop resistance to multiple exposures of anti‐HER2 therapies. New strategies need to be developed for a more accurate selection of patients who can benefit from these treatments. While much is known about HER2 biology, the biochemical impact of HER2 on breast cancer cell metabolism is not well understood. The goal of this project is to better understand HER2 metabolomics and associated molecular mechanisms using Nuclear Magnetic Resonance (NMR) spectroscopy. A series of eight different breast cancer cell lines ranging from low to high HER2 expression were adapted to a common media with negligible morphological change or alteration in HER2 expression levels. Metabolite profiles for experimental replicates of each cell line were determined using high‐resolution 1H‐NMR, principal components, and pathway analyses. This study identified pathways associated with metabolic reprogramming of nonessential amino acids. Specifically, pathway and enrichment analyses identified glycine and serine as the most significant which are known pathways supporting one‐carbon metabolism. Furthermore, the data showed an increased consumption of glutamate, which is necessary for energy production and nucleotide biosynthesis. These pathways support the altered metabolism and inefficient production of energy via aerobic glycolysis in breast cancer cells promoting tumor growth and progression. Ultimately, we hope to exploit these discoveries for metabolic targeting as a therapeutic strategy against HER2‐positive breast cancer cells.Support or Funding InformationNIH grant 5SC3GM122630 to JAB
Scaffolding Protein IQ Motif Containing GTPase Activating Protein 2 Regulates Liver Metabolic Homeostasis
Authors: Sen Anuhna , Youssef Sara , Wendt Karen , Schmidt Valentina , Anakk Sayeepriyadarshini 
Published: April 2020

The liver plays a central role in maintaining metabolic flexibility by regulating signaling pathways. Disruption in this process can lead to metabolic disease, such as obesity and cardiovascular risk. Scaffold proteins are well known to coordinate many signaling cascades. Since IQ Motif Containing GTPase Activating Protein 2 (IQGAP2) is a highly expressed scaffold protein in the liver, we examined its role in regulating metabolic programs. Adult wild type and Iqgap2−/− mice were fed ad libitum or fasted for 24 hours (9.00 am, or ZT4) and both male and female mice were used in this study to determine for sex differences in metabolism. We examined their gross morphology, performed serum biochemical assays and quantified expression of key enzymes regulating metabolic pathways including fat and carbohydrates. We found deficiency of IQGAP2 did not alter gross morphological features, liver‐to‐body weight ratio or white adipose tissue (WAT) weight. Iqgap2−/− male mice did not show pronounced differences in metabolic gene expression or fuel accumulation. In contrast, Iqgap2−/− female mice showed decreased expression of gluconeogenic, ketogenic and fatty acid oxidation genes. This basal decrease was compensated upon fasting with higher induction of ketogenic and gluconeogenic programs in Iqgap2−/− female mice. Our preliminary data suggest female Iqgap2−/− mice have defective nutrient storage. The two main hepatic fuel stores are glycogen and triglycerides. While hepatic triglycerides were not altered in female Iqgap2−/− mice, glycogen accumulation was significantly reduced. Histological analysis of hepatic glycogen accumulation showed distinct pericentral zonation which was apparent in the fed state. Consistent with this result, female Iqgap2−/− livers displayed reduced expression of genes regulating glycogen synthesis and degradation. Taken together, these data suggest a novel role for IQGAP2 in regulating hepatic fuel storage, especially glycogen levels.Support or Funding InformationIllinois Campus Research Board and American Cancer Society
The C‐terminal Domain of AAGAB Mediates Tetramerization and Stability
Authors: Yin Qian , Tian Yuan , Crisman Lauren Christine , Wang Bing , Li Suzhao , Shen Jingshi 
Published: April 2020

In clathrin‐mediated endocytosis (CME), the coat protein clathrin relies on
cargo adaptors such as the heterotetrameric AP2 adaptor complex to recruit cargo proteins to endocytic sites. It was previously thought that multimeric cargo adaptors such as AP2 adaptor assemble spontaneously. However, we recently discovered that AP2 adaptor assembly is an ordered process controlled by alpha and gamma adaptin binding protein (AAGAB). Without the assistance of AAGAB, AP2 adaptor fails to form, leading to CME defects. Heterozygous AAGAB mutations cause human diseases such as puncate palmoplantar keratoderma type 1 (PPKP1), a skin disorder characterized by lesions on palms and soles. However, the biochemical property and structure of AAGAB remain to be established. AAGAB is a soluble protein with two conserved regions – an N‐terminal G protein‐like domain (G domain) and a C‐terminal domain not similar to any known protein domain. We found that the full‐length AAGAB exists entirely as a homotetramer in solution and the tetramerization is mediated by its C‐terminal domain. Using X‐ray crystallography, we solved the atomic structure of the C‐terminal tetramerization domain (TD). The structure unveiled that the TD tetramer is formed by dimerization of two dimers. Furthermore, we observed that the expression level of the TD‐deficient AAGAB mutant (AAGABdeltaTD) in cells was drastically reduced compared to full‐length AAGAB. Together, our findings demonstrated that the TD mediates AAGAB tetramerization and is required for the stability of free AAGAB proteins prior to AP2 binding. These data also provide the first molecular explanation for
the disease‐causing nonsense mutations found on AAGAB lacking the C‐terminal tetramerization domain.Support or Funding InformationFlorida State University startup funds to Yin; NIH/NIDDK R56 DK095367 to Shen; NIH/NIAID R21 AI128443 to Shen and Li.
Sputtering of Nanometric Films As a Tool to Enhance Antimicrobial Properties
Authors: Beckford Daihlia , Ward Zack , Phillips Jacey , Repa Kristen Stojak , Miller Casey , Michel Lea Vacca 
Published: April 2020

Sputtering is a high vacuum physical vapor deposition technique during which atoms are excited from the surface of a target by a plasma. The energized atoms create a thin layer (thin film) on the surface of a chosen substrate material. Film thickness is determined by the deposition time (longer time thicker films). Our project aims to enhance the antimicrobial properties of a variety of textiles, polymers, and other surfaces by sputtering a nanometric film of different metals such as copper (Cu), gold (Au), and silver (Ag). Here, we describe our efforts to develop and optimize our experimental methods to assess the antibacterial properties of our sputtered surfaces, using Escherichia coli (E. coli) as the bacterial source. Preliminary data from our experiments suggest that our sputtering technique, which employs a “seed layer” of metal, successfully deposits a metallized film onto some surfaces, and that some metals are more easily washed off those surfaces than others. We also describe our different approaches toward the assessment of the antibacterial properties of the metallized surfaces.Support or Funding InformationSchool of Chemistry and Materials Science, Rochester Institute of Technology
Sulfation of endothelial heparan sulfate mediates Staphylococcus aureus induced hepatotoxicity
Authors: Golden Gregory James , Toledo Alejandro Gómez , Cuello Hector , Nizet Victor , Esko Jeffrey 
Published: April 2020

Sepsis is defined as life‐threatening organ dysfunction caused by a dysregulated host response to infection and vascular pathology. Understanding the linkage of vascular pathology to organ failure could provide a strategy for treating the disease. Heparan sulfate proteoglycans are critical components of the vascular endothelial glycocalyx and play a key role in sepsis outcome in mice. To study how heparan sulfate modulates organ‐specific vascular pathology during sepsis, we induced Staphylococcus aureus sepsis in Ndst1f/fTie2Cre mice, which express under‐sulfated heparan sulfate in endothelial and myeloid tissues. Hepatic coagulopathy and infarction induced by S. aureus was reduced in Ndst1f/fTie2Cre mice, but not in Ndst1f/fLysMCre or Ndst1f/fPF4Cre mice. To examine how modifying heparan sulfate structure mitigates hepatic damage, pro‐inflammatory neutrophil trafficking and behavior was monitored in the liver during sepsis. Ndst1f/fTie2Cre mice contained fewer extravasated neutrophils in the liver early in sepsis. Intravital microscopy in the liver further verified that Ndst1f/fTie2Cre mice have neutrophil trafficking deficiencies after both sterile and septic injury. Further studies will focus on the role of heparan sulfate in neutrophil trafficking and downstream neutrophil‐driven vascular dysfunction in sepsis.Support or Funding InformationThis work is supported by NIH P01 HL131474 and the UCSD Microbial Sciences Graduate Research Initiative Fellowship.
A Realistic View of Proteins : Solution Scattering at the Life Sciences X‐ray Scattering Beamline (LIX)
Authors: Byrnes James , Chodankar Shirish , Lazo Edwin , Qian Kun , Shea-McCarthy Grace , Antonelli Stephen , Yang Lin 
Published: April 2020

Small angle X‐ray scattering is an important tool that reveals the behavior of proteins in their native environment. The scattering pattern produced from a SAXS experiment represents an average of all conformations present in the solution. Since most proteins in our body function in a solution environment, SAXS data can reveal additional biologically relevant states of a molecule or molecular complex. Furthermore, SAXS provides data about the shape and size of the molecule of interest and is a complementary technique to other structural methods. Combining SAXS data with X‐ray crystallography, NMR or Cryo‐EM can lead to a more robust understanding of the species being studied. High quality SAXS data can be used to build ab initio low resolution bead models, which provide a 3D envelope of your molecule. Here at the life sciences X‐ray scattering beamline (LiX), we collect data by flowing sample through the X‐ray path in a 3 channel flow cell. As the sample enters the X‐ray path, the simultaneous collection of SAXS and wide angle X‐ray scattering (WAXS) is obtained via a 3 detector setup. The detector setup has a q‐range of 0.006Å−1 to 3.2 Å−1, primarily to obtain scattering data from the water peak to normalize buffer subtraction. Automated sample handling at LiX allows for collection of up to 360 samples, packaging of data and processing. One challenge in obtaining high quality SAXS data is preparation of pure, monodisperse samples. For these complicated samples that have multiple species, our in‐line HPLC system seamlessly integrates with our beamline for SEC‐SAXS (size‐exclusion chromatography‐SAXS), and increases the chances of obtaining a more pure, monodisperse sample immediately before data collection.Support or Funding InformationLiX beamline is part of the Life Science Biomedical Technology Research resource, co‐funded by the National Institute of General Medical Sciences (NIGMS) under grant P41 GM111244 and by the DOE Office of Biological and Environmental Research under grant KP1605010, with additional support from NIH under grant S10 OD012331. The operation of NSLS2 is supported by US Department of Energy, Office of Basic Energy Sciences, under contract No. DE‐SC0012704.
Adrenergic signaling‐induced ultrastructural strengthening of intercalated discs via PG is crucial for positive adhesiotropy in murine cardiomyocytes.
Authors: Yeruva Sunil , Kempf Ellen , Egu Desalegn Tadesse , Kugelmann Daniela , Waschke Jens 
Published: April 2020

Intercalated discs (ICD), which connect adjacent cardiomyocytes, are composed of desmosomes, adherens junctions (AJ) and gap junctions (GJ). Previous data demonstrated that adrenergic signaling enhances cardiac myocyte cohesion, referred to as positive adhesiotropy, via PKA‐mediated phosphorylation of plakoglobin (PG). Here, we further investigated the role of PG in adrenergic signaling‐mediated ultrastructural changes in ICD of cardiomyocytes. Quantitative transmission electron microscopy analysis of ICD demonstrated that cAMP elevation caused significant elongation of area composita and thickening of the ICD plaque, paralleled by enhanced cardiomyocyte cohesion, in WT but not PG‐deficient cardiomyocytes. STED microscopy analysis supported that cAMP elevation ex vivo enhanced Dsg2 and N‐cadherin (N‐cad) staining in ICDs of WT but not PG‐deficient cardiomyocytes. For dynamic analyses we utilized HL‐1 cardiomyocytes, in which cAMP elevation induced translocation of desmoglein‐2 (Dsg2) and PG but not of N‐cad to cell junctions. Nevertheless, depletion of N‐cad but not of Dsg2 resulted in a decrease in basal cell cohesion whereas positive adhesiotropy was abrogated in monolayers depleted for either Dsg2 or N‐cad. Moreover, AFM experiments showed that the cAMP elevation‐induced increase in Dsg2 binding events close to cell junctions was abolished by depletion of either Dsg2 or N‐cad. Our data demonstrate that in murine cardiomyocytes, adrenergic signaling via PG enhances N‐cad and Dsg2 in the ICD leading to strengthening of ICDs and thereby positive adhesiotropy.Support or Funding InformationThis work was supported by the Ludwig‐Maximilians‐Universität Munich with the Föfole program; and the Deutsche Forschungsgemeinschaft DFG grant number WA2474/11‐1 to Jens Waschke
Inflammation and Immune Markers in Psychological Health
Authors: Scholl Jamie L. , King Zach , Potter Kari , DeCino Daniel A. , Hertel Danielle , Pearson Kami , Graack Eric , Zeng Erliang , Brown-Rice Kathleen , Forster Gina L. 
Published: April 2020

There has been a growing emphasis on the relationship between inflammatory responses, gut microbiota and the immune system, and their influence on mental health outcomes. Stress can influence the composition of gut microbiota which, in turn, influences the central nervous systems response to future stressors. C‐Reactive Protein (CRP) is a biomarker for inflammation that has been linked to depression, anxiety disorders and PTSD that is released in response to pro‐inflammatory cytokines. The goal of this study was to test a direct association between behavior profiles such as anxiety and depression, the composition of the gut microbiota, basal levels of CRP and cytokines, cortisol and genetic predisposition. Measures of anxiety, depression, PTSD, substance use and overall health were obtained from adults with no recent illness or surgery, or exposure to antibiotics. Biological measures focused on pro‐ and anti‐inflammatory cytokines from plasma, cortisol levels from hair, and gut microbiome; as well as single nucleotide polymorphisms (SNPs) were assayed. Using factor analysis we determined a shared mental health panel including depression, anxiety and PTSD measures that was negatively correlated to related cytokines. Regression modeling was used to related quantitative traits to demographic and genetic factors, revealing significant relationships between inflammatory markers and mental health. Understanding these relationships will help to determine direct future research in assessing causal direction between all of these factors that can promote mental well‐being, and could in the future be used as biomarkers for vulnerability to mental illness, and even help re‐think how we treat mental health disorders.Support or Funding InformationThe project described was supported by the National Institute Of General Medical Sciences, 1U54GM115458‐01; The Center for Brain and Behavior Research (CBBRe), USD; Basic Biomedical Sciences, USD; and the South Dakota Governor’s Center for Genetics and Behavioral Health.