# ent

id: cord-000933-nn9gj0z1
type: PROTEIN
entity:
  Millipore anti F monoclonal ab43812 abcam anti P rabbit polyclonal 3 V Biosciences Menlo Park USA anti ZO 1 goat anti mouse goat anti rabbit


id: cord-018771-xqlb74px
type: DNA
entity:
  Zubereitungen zur Verfügung Gewaschene Erythrozytenkonzentrate Diese können dann transfundiert werden


id: cord-018771-xqlb74px
type: CHEMICAL
entity:
  Patienten mit einer chronischen Anämie sollten daher bei einer Hämoglobinkonzentration unter 7 8 g dl Hämatokrit 21 24 transfundiert werden Evidenzgrad 1 C sofern andere Maßnahmen wie


id: cord-018771-xqlb74px
type: CHEMICAL
entity:
  Patienten beatmungspflichtig Therapeutisch stehen die Sicherung einer adäquaten Oxygenierung sowie eine


id: cord-018771-xqlb74px
type: DISEASE
entity:
  durch die Antikörperbildung bei einer Schwangerschaft mit einem Rhesus positiven Kind ebenfalls zu einer Rhesusunverträglichkeitsreaktion kommen kann


id: cord-258678-0atfsivf
type: ORG
entity:
  Systematic Investigation of Optimal Aptamer Immobilization for Proteinâ Microarray Applications Minor


id: cord-001123-n2e4s7bu
type: WORK_OF_ART
entity:
  The Soluble Form of the EIAV Receptor Encoded by an Alternative Splicing Variant Inhibits EIAV Infection of Target Cells


id: cord-001123-n2e4s7bu
type: DISEASE
entity:
  human immunodeficiency virus HIV 1 simian immunodeficiency virus SIV and feline immunodeficiency virus FIV


id: cord-002076-7t4d4vvo
type: DISEASE
entity:
  human immunodeficiency virus type 1 activity Structure based discovery of Middle East respiratory syndrome coronavirus


id: cord-307067-cpc1yefj
type: PROTEIN
entity:
  interleukin 2 IL 2 IL 6 IL 8 IL 18 monocyte chemotactic protein 1 MCP 1 macrophage inflammatory protein 1a MIP 1


id: cord-307067-cpc1yefj
type: PROTEIN
entity:
  granulocyte colony stimulating factor G CSF granulocyte macrophage CSF interferon IL 1 IL 1 receptor antagonist IL 2 IL 4 IL 5 IL 6 IL 8 IL 10 IL 12 23 p40 IL 13 IL 15 IL 17 IL 18 MCP 1


id: cord-102184-8u73adnk
type: GGP
entity:
  bioRxiv DOI 10 1101 2020 02 07 935924 sha 12970fd08012cac78df72cea4a9cda70582acf0b doc_id 102184 cord_uid 8u73adnk The ability to integrate


id: cord-319517-denczc6t
type: PROTEIN
entity:
  Aequorea victoria green fluorescent protein Science Improved green fluorescence Fluorescence proteins


id: cord-001270-l8aa9cl3
type: PROTEIN
entity:
  GFP JL8 1 5000 Clontech α tubulin mouse monoclonal antibody 1 8 000 Sigma HA high affinity anti HA antibody rat monoclonal 1 1 000 Roche cat 11867423001


id: cord-315483-l6dm82pp
type: DNA
entity:
  SCIentIFIC REPORtS 2018 8 6794 DOI 10 1038 s41598 018 24905 y duck and human IFIT5 Fig 1C One of the structural hallmarks


id: cord-315483-l6dm82pp
type: CELL_LINE
entity:
  42 Immortalized chicken fibroblasts DF 1 human embryonic kidney cells 293T HEK 293T and Madin Darby canine kidney MDCK cells


id: cord-275307-d7htyfcl
type: GGP
entity:
  n 61 SOX n 178 B Frequency distribution histogram of log likelihood scores for the 20 nt surrounding


id: cord-317142-qd61qvch
type: PROTEIN
entity:
  N and C terminal pro sequences Q 1N Q306N are 35 723 and 40 526 Da respectively However under some conditions the mobility of GFP fused proteins


id: cord-317142-qd61qvch
type: DNA
entity:
  oligonucleotides 5 CAATCACTTCTGCTGTTCTGA ACAGTGGTTTTAGGAAAATGGC 3 and 5 GCCATT TTCCTAAAACCACTGTTCAGAACAGCAGAAGTGA TTG 3


id: cord-317142-qd61qvch
type: DNA
entity:
  Q30 9N and 5 GGTTCTTTCCTTAATGGATCAGCTGGTA GTGTTGGTTT TAAC 3 and 5 GTTAAAACCAACAC TACCAGCTGATCCATTAAGGAAAGAACC 3


id: cord-317142-qd61qvch
type: PROTEIN
entity:
  creatine phosphate 250 lgÁmL À1 creatine kinase 0 64 mM 3 5 cyclic AMP 68 lM L 5 formyl 5 6 7 8 tetrahydrofolic acid 175 lgÁmL À1 E


id: cord-317142-qd61qvch
type: CHEMICAL
entity:
  N terminal processing site for WT based on their molecular weights estimated by SDS PAGE Fig 1B Measuring the activity of 3CL


id: cord-317142-qd61qvch
type: CHEMICAL
entity:
  N and C terminal processing sites were estimated Table 2 from the data in Table 1 Based on the assumption that between any dimeric protomer species the monomer dimer exchange rate is so fast that the population of each dimer at each time is proportional to the population of the protomers forming the dimer the maturation process was simulated as shown in Fig 5A This assumption is


id: cord-317142-qd61qvch
type: CHEMICAL
entity:
  N terminal processing site of a protomer was cleaved by the active site of the other protomer cis cleavage After cleavage of both N termini of the protomers rearrangement of the protomers occurs to form the same dimeric structure


id: cord-332881-mkm4ygh6
type: PROTEIN
entity:
  human signaling lymphocytic activation molecule SLAM CD150 Respiratory syncytial virus infection of monocyte


id: cord-346554-a98pjtxs
type: CELL_LINE
entity:
  CCL 2TM Ceropithecus aethiops epithelial kidney Vero ATCC CCL 81 human epithelial HeLa contaminant carcinoma HEp 2


id: cord-345651-admlzeu4
type: ORG
entity:
  The N Terminal Domain of Spike Protein Is Not the Enteric Tropism Determinant for Transmissible Gastroenteritis Virus


id: cord-319842-4mnaicki
type: DISEASE
entity:
  picornaviruses poliovirus rhinovirus 2 or rhinovirus 14 or transfected with plasmids that express poliovirus proteins 2BC and 3A see Figures 2


id: cord-353467-wbtzvm4i
type: CHEMICAL
entity:
  peptide N glycosidase F PNGase F which converted the gp27 form of S to its faster migrating p24 form The GFP S and S YFP


id: cord-334855-s0ci3r8w
type: ORG
entity:
  Bovine Viral Diarrhoea Virus BVDV Replication Characterization of Dengue Virus Resistance to Brequinar in Cell Culture


id: cord-348799-qu4zin3o
type: CELL_TYPE
entity:
  human peripheral blood T lymphocyte Jurkat Jurkat human peripheral blood monocytic THP 1 cells THP 1


id: cord-348799-qu4zin3o
type: DNA
entity:
  ORF 5 GATCACTAATACGACTCACTATAGGTGGG CCTCAAAGGGTGAGGTTTTAGAGCTAGAAAT 3 In vitro Transcription T7 Kit 6140 TaKaRa


id: cord-348799-qu4zin3o
type: DISEASE
entity:
  Venezuelan equine encephalitis viruses Rhabdoviridae Vesicular Stomatitis virus VSV Orthomyxoviridae Influenza virus Retroviridae Human Immunodeficiency Type 1 virus Hepadnaviridae Hepatitis B


id: cord-342189-ya05m58o
type: DISEASE
entity:
  influenza A encodes a protein that modulates mRNA splicing to degrade the mRNA that encodes RIG I Kochs


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  H1 A M Polyanichko 1 H Wieser 2 1 Faculty of Physics Saint Petersburg State University Russia 2 Dept


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  antitumor nonclassical platinum complexes O Vrana V Kohoutkova V Brabec Institute of Biophysics AS CR Královopolská 135 CZ 61265


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  CYP450 L S Goto 2 C O Hokka 1 J F Lima 2 O R Nascimento 2 A P U Araújo 2 1 Grupo de Engenharia Bioquímica UFSCar BR 2


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  Complex I A L Maniero 1 C Bergamini 2 M Bortolus 1 R Fato 2 S Leoni 2 G Lenaz 2 1 Università degli Studi di Padova Padova Italy 2 Università degli studi di Bologna Bologna Italy Complex I NADH Dehydrogenase


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  arXiv 0805 0696v1 S Soria 1 F Quercioli 3 R Mercatelli 3 F Bianco 2 I Cacciari 2 G Righini 2 1 Centro Studi e Ricerche E Fermi P Del Viminale 2 00184 Rome Italy 2


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  plague F1 capsular antigen A Soliakov 1 J R Harris 1 M R Hicks 2 A Rodger 2 R Woody 3 A Watkinson 4 J H Lakey 1 1 Cell


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  membrane proteins C V Kulkarni 3 O Ces 1 S Iwata 2 R H Templer 1 1 Chemical Biology Centre and Department of Chemistry Imperial College London United Kingdom 2 Division of Molecular


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  lambda bacteriophage epigenetic switch L Finzi Physics Department emory University 400 dowman Dr Atlanta GA 30322 USA


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  topoisomerase II activity H Seidel 1 T Klose 2 H Lilie 2 C G Hübner 1 1 Institute of Physics Ratzeburger Allee 160 23538 Lübeck Germany 2 Institute of Biochemistry and Biotechnology Kurt Mothes Str 3 06120 Halle Germany One essential element


id: cord-004534-jqm1hxps
type: DNA
entity:
  human MEF2D gene W Zhou 1 L Ying 2 1 Molecular Medicine National Heart and Lung Institute Imperial College London London SW7 2AZ UK 2 Chemical Biology


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  OmpF channel E Hajjar 1 A Kumar 1 M Winterhalter 2 P Ruggerone 1 M Ceccarelli 1 1 Department of Physics University of Cagliari Italy 2


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  protein N Maghelli 1 V Krstic 2 N Pavin 2 F Julicher 2 I Tolic Norrelykke 1 1 MPI CBG Dresden Germany 2 MPI PKS Dresden Germany


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  integrin αIIbβ3 M Rocco 1 C Rosano 1 J W Weisel 2 D Horita 3 R R Hantgan 3 1 Istituto Nazionale per la Ricerca sul Cancro IST Genova Italy 2


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  redox cofactor L Erokhova 1 P Kügler 2 P Pohl 1 1 Institute of Biophysics Johannes Kepler University Linz Austria 2


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  cation channels Y Sawada 1 M Murase 2 M Sokabe 1 1 Dept Physiol Nagoya Univ Grad Sch Med Nagoya Japan 2


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  Na K ATPase C H Chang 1 T Y Tsong 2 1 Institute of Physics National Chiao Tung university Physics Division National Center for Theoretical Sciences Hsinchu Taiwan 2 Institute of Physics Academy of Sciences Taipei 115 Taiwan Na K ATPase


id: cord-004534-jqm1hxps
type: CELL_TYPE
entity:
  permeabilized skeletal muscle fibres D S Ushakov D Ibanez Garcia T G West P M W French M A Ferenczi Imperial College London UK


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  extracellular matrix molecule hyaluronic acid modulates L type voltage dependent Ca2 channels E Dvoretskova 1 G Kochlamazashvili 2 O Bukalo 2 C Henneberger 3 D Rusakov 3 M Schachner 2 A Dityatev 1 1 Italian Institute of Technology Genova Italy 2


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  colicin translocon C L Johnson 1 A Solovyova 1 P Callow 2 S A Holt 3 L A Clifton 3 K Weiss 4 J H Lakey 1 1


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  human 3 phosphoglycerate kinase P Lallemand 1 J Rouhana 1 L Chaloin 1 B Roy 2 S Arold 3 T Barman 1 C Lionne 1 1 CPBS UMR 5236 4 bd Henri IV


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  Green Fluorescent Protein G Jung Biophysical Chemistry Saarland University 66123 Saarbruecken Germany Biotechnological design of the Green Fluorescent Protein GFP


id: cord-004534-jqm1hxps
type: PROTEIN
entity:
  J Strömqvist 1 S Johansson 1 Y Ohsugi 3 K Andersson 2 L Xu 1 M Kinjo 3 P Höglund 2 J Widengren 1 1 Experimental biomolecular Physics KTH Stockholm Sweden 2


id: cord-004534-jqm1hxps
type: GGP
entity:
  Alzheimer s disease Some diabetes drugs appear to decrease the cognitive decline associated with Alzheimer s


id: cord-004534-jqm1hxps
type: GGP
entity:
  human 3 phosphoglycerate kinase P Lallemand 1 J Rouhana 1 L Chaloin 1 B Roy 2 S Arold 3 T Barman 1 C Lionne 1 1


id: cord-004534-jqm1hxps
type: ORG
entity:
  Institute for Problems of Cryobiology and Cryomedicine National Academy of Sciences of Ukraine Kharkov Ukraine


id: cord-004534-jqm1hxps
type: ORG
entity:
  Physics Department University of Genoa Italy 2 Biochemical Sciences Department University of Florence Italy Short


id: cord-004534-jqm1hxps
type: ORG
entity:
  Molski Adam Mickiewicz University Laboratory for Dynamics of Physicochemical Processes Poznan Poland RNAzymes


id: cord-004534-jqm1hxps
type: ORG
entity:
  Department of Physics Faculty of Science Messina University Italy 2 Department of Physics Faculty of Science Perugia University Italy Important


id: cord-004534-jqm1hxps
type: ORG
entity:
  Chemistry University of Oxford Oxford U K 2 Department of Electronics Computer Science and Systems University of Bologna Bologna Italy Potassium


id: cord-004534-jqm1hxps
type: ORG
entity:
  Institute of Physics National Chiao Tung university Physics Division National Center for Theoretical Sciences Hsinchu


id: cord-004534-jqm1hxps
type: ORG
entity:
  Department of Physics University of Warwick U K 2 Department of Chemistry University of Warwick U K Information


id: cord-004534-jqm1hxps
type: ORG
entity:
  Biotechnology Jagiellonian University 2 Department of Pharmacodynamics Medical College Jagiellonian University


id: cord-004534-jqm1hxps
type: LAW
entity:
  A Isvoran 1 E Quiniou 2 C Craescu 2 L Mouawad 2 1 West University of Timisoara Department of Chemistry Pestalozzi


id: cord-004534-jqm1hxps
type: WORK_OF_ART
entity:
  A Martino D Crane I M Feavers B Bolgiano Division of Bacteriology National Institute for Biological Standards and Control Potters


id: cord-004534-jqm1hxps
type: CHEMICAL
entity:
  E Del Favero 1 G Di Fede 2 F Tagliavini 2 M Salmona 3 L Cantù 1 1 University of Milan Segrate Italy 2 Carlo


id: cord-004534-jqm1hxps
type: CHEMICAL
entity:
  L D Alfonso 1 M Collini 1 M Caccia 1 G Tallarida 2 S Caprioli 2 G Chirico 1 1 Dipartimento di Fisica Università di Milano Bicocca Italy 2 Laboratorio


id: cord-004534-jqm1hxps
type: CHEMICAL
entity:
  1 D Magatti 1 B Cardinali 2 A Profumo 2 M Rocco 2 1 Dipartimento di Fisica e Matematica Università dell Insubria


id: cord-274210-uaaj66xq
type: DISEASE
entity:
  abnormal nerve cell morphology unnatural protein accumulation vesicular transport or neuronal gene transcription disorders


id: cord-023026-2r84ndzv
type: CELL_LINE
entity:
  x3 Balanced OB Deficient OD or Supplemented OS rats We have evaluated synaptic efficacy and plasticity electrophysiological recording astroglial regulations glutamate uptake and GFAP expression neuronal markers glutamate transporters


id: cord-023026-2r84ndzv
type: PROTEIN
entity:
  oligomeric proteins hamster polyomavirus major capsid protein VP1 human metapneumovirus nucleocapsid protein


id: cord-023026-2r84ndzv
type: PROTEIN
entity:
  vascular adhesion proteins ICAM 1 VCAM 1 ITGA9 proangiogenic cytokines PlGF Il 8 HB EGF metalloproteinase MMP 9


id: cord-023026-2r84ndzv
type: CL
entity:
  astroglial CPE and SgIII were analyzed under basal and stimulated conditions in primary cultures We found that exposure of astrocytes


id: cord-023026-2r84ndzv
type: CL
entity:
  neuronal cells 1 2 3 4 5 Atomic force microscopy is a powerful tool not only to study the morphology in terms of high resolution imaging and roughness measurements but also to map mechanical and adhesive properties of the sample cells


id: cord-023026-2r84ndzv
type: CL
entity:
  astroglial and microglial processes The deprivation also resulted in gradual increase of mitochondrial


id: cord-023026-2r84ndzv
type: GGP
entity:
  AMPA receptors evokes Ca 21 entry in isolated Bergmann glia processes and Ca 21 dependent vesicular glutamate


id: cord-023026-2r84ndzv
type: LAW
entity:
  L Vargova 1 M Cicanic 1 2 E Sykova 1 2 1 Charles University 2nd Faculty of Medicine Prague Czech Republic 2 Institute of Experimental Medicine v


id: cord-023026-2r84ndzv
type: WORK_OF_ART
entity:
  A Grimaldi G D Alessandro C Lauro M Catalano C Limatola Sapienza University of Rome Rome Italy Glioblastoma GBM


id: cord-023211-kt5gt26t
type: DNA
entity:
  poly ubiquinated F508 CFTR Wang Y 1 Jiang Y 1 Zhu N 1 Feng X 1 Yang H 1 2 Ma T 1 2 1 Membrane Channel Research Laboratory Northeast Normal University Changchun China 2 Biopharmaceutical Center Liaoning Normal University Dalian China Deletion


id: cord-023211-kt5gt26t
type: PROTEIN
entity:
  NIH 5 RO1HL074175 06 and 5 T32 DK007301 29 1 1 Physiology Dartmouth Medical School Hanover NH USA 2 Microbiology and Immunology Dartmouth Medical School Hanover NH USA


id: cord-023211-kt5gt26t
type: CELL_LINE
entity:
  NIH 5 RO1 HL074175 Hamai H 1 Keyserman F 1 Worgall T S 1 2 1 Pathology Columbia University New York NY USA 2 Pediatrics Columbia University New York NY USA Cystic fibrosis CF


id: cord-023211-kt5gt26t
type: CELL_LINE
entity:
  NB DGJ Human bronchial IB3 1 CFTR F508del W1282X CuFi 1 CFTR F508del F508del and NuLi 1 CFTR w t cells


id: cord-023211-kt5gt26t
type: CELL_LINE
entity:
  Ad FOXJ1 EGFP treated cultures 0 6 0 2 µA cm2 n 9 The increase in Isc was blocked by INH172 an inhibitor of CFTR mediated secretion Under these conditions Ad


id: cord-023211-kt5gt26t
type: CELL_LINE
entity:
  North Carolina Hospitals Chapel Hill NC USA 4 Dept Pathology and Laboratory Medicine University of North Carolina Hospitals Chapel Hill NC USA


id: cord-023211-kt5gt26t
type: GGP
entity:
  Tx 18 2 1 7 g L vs 22 2 3 9 g L p 0 026 ALP was higher 1 month post lung Tx 389 5 vs 137 9 p 0 033 and ALT


id: cord-023211-kt5gt26t
type: FAC
entity:
  Chapel Hill Chapel Hill NC USA 3 Center for Environmental Health and Susecptibility Univeristy of North Carolina


id: cord-023211-kt5gt26t
type: WORK_OF_ART
entity:
  Pediatrics National Jewish Medical and Research Center Denver CO USA 2 Integrated Department of Immunology National Jewish Medical and Research Center Denver CO USA 3 Department of Medicine Cystic Fibrosis Pulmonary Research and Treatment center


id: cord-023211-kt5gt26t
type: ORG
entity:
  University of North Carolina Hospitals Chapel Hill NC USA 3 School of Public Health University of North Carolina Hospitals Chapel Hill NC USA 4 Dept Pathology and Laboratory Medicine University of North Carolina Hospitals Chapel Hill NC USA Introduction Proportion


id: cord-023211-kt5gt26t
type: WORK_OF_ART
entity:
  Pediatrics Women Children s Hospital Buffalo NY USA 2 Pediatrics University of Rochester Rochester NY USA Background Airway


id: cord-023211-kt5gt26t
type: ORG
entity:
  Physical Role Vitality Emotion Social Body Image Eating Treatment Burden Health Status Weight Respiratory Symptoms and Digestion


id: cord-023211-kt5gt26t
type: ORG
entity:
  3 School of Medicine University of Pittsburgh Pittsburgh PA USA 4 Dept of Medicine Clinic Sattelduene


id: cord-023211-kt5gt26t
type: ORG
entity:
  Whittaker L A Tilluckdharry L Christian R Medicine University of Vermont Burlington VT USA OBJECTIVE Bone


id: cord-023211-kt5gt26t
type: CHEMICAL
entity:
  bisaminomethylbithiazole Corr 4a N 2 5 chloro 2 methoxyphenylamino 4 methyl 4 5 bithiazol 2 yl benzamide


id: cord-023211-kt5gt26t
type: DISEASE
entity:
  Aspergillus fumigatus Aspergillus versicolor Pichia guilliermondii Rhodotorula sp Trichosporon sp Aureobasidium pullulans Fusarium sp Mucor hiemalis Bionectria ochroleuca Gibberella pulicaris


id: cord-023211-kt5gt26t
type: CHEMICAL
entity:
  Feranchak A P 1 1 Department of Pediatrics UT Southwestern Medical Center Children s Medical Center Dallas TX USA 2 Medicine UT Southwestern Dallas TX USA


id: cord-257167-rz4r5sj7
type: CELL_LINE
entity:
  cat V1 Ayako Ishikawa 1 Satoshi Shimegi 2 Hiroyuki Kida 3 Hiromichi Sato 2 1 Grad Sch Front Biosci Osaka Univ Osaka Japan 2 Grad Sch Med Osaka Univ Japan 3 Grad Sch Eng Sci Osaka Univ Japan


id: cord-257167-rz4r5sj7
type: DNA
entity:
  rat Norio Ishizuka Yoshitomo Umitsu Department of Brain Structure Tokyo Metropolitan Institute for Neuroscience Tokyo Japan Longitudinal fiber systems


id: cord-257167-rz4r5sj7
type: CELL_LINE
entity:
  rat pituitary Katsutoshi Taguchi 1 Haruko Kumanogoh 2 Shun Nakamura 2 Seiji Miyata 3 Shohei Maekawa 1 1 Department of Biosystems Science Kobe University Kobe Japan 2 Division of Biochemistry and Cellular Biology National Institute of Neuroscience


id: cord-257167-rz4r5sj7
type: DNA
entity:
  CA3 region Ryozo Aoki 1 Hiroshi Wake 2 Hitoshi Sasaki 1 Kiyokazu Agata 3 1 Dept Physiol Biosignal Osaka Univ Grad Sch Med Suita Japan 2 Dept Elec Eng Elec Col Industri Tech Amagasaki Japan 3 Dept Biophys Kyoto Univ Grad Sch Sci Kyoto Japan


id: cord-257167-rz4r5sj7
type: PROTEIN
entity:
  Scrapper protein PS2P A003 Presynaptic membrane potential dependent regulatory mechanism of transmitter release Tetsuya Hori Tomoyuki Takahashi Department of Neurophysiology University of Tokyo Graduate School of Medicine Tokyo Japan In simultaneous pre and postsynaptic recordings at the calyx of Held


id: cord-257167-rz4r5sj7
type: PROTEIN
entity:
  JAK STAT Hiroko Inoue 1 Haruhisa Kawasaki 2 Ritsuko Fujii 2 Tohru Yoshioka 2 Kazunori Sango 3 Toshihiko Kadoya 4 Hidenori Horie 2 1 Sch


id: cord-257167-rz4r5sj7
type: PROTEIN
entity:
  collapsin response mediator protein 2 CRMP 2 Masumi Iketani 1 Yanai Shigeki 1 Fumio Nakamura 1 Yoshio Goshima 1 2 Kohtaro Takei 1 2 1 Dept of Mol Pharmacol Neurobiol Yokohama City Univ Grad Sch


id: cord-257167-rz4r5sj7
type: PROTEIN
entity:
  protein tyrosine phosphatase receptor type O Takafumi Shintani 1 2 Masaru Ihara 1 2 Hiraki Sakuta 1 2 Hiroo Takahashi 1 2 Ikuko Watakabe 1 Masaharu Noda 1 2 1


id: cord-257167-rz4r5sj7
type: PROTEIN
entity:
  EGFP GluR1 Hamid Gholamipour 1 Shirin Babri 2 Khameneh Saied 3 1 Department of Physiology University


id: cord-257167-rz4r5sj7
type: DNA
entity:
  SERT Junichi Kitanaka 1 Nobue Kitanaka 1 Tomohiro Tatsuta 1 2 Yoshio Morita 2 Motohiko Takemura 1 1 Department of Pharmacology Hyogo College of Medicine Nishinomiya Japan 2 Department of Neuropsychiatry Hyogo College of Medicine Nishinomiya Japan


id: cord-257167-rz4r5sj7
type: PROTEIN
entity:
  proteins neuronal nuclear antigen NeuN glial fibrillary acidic protein GFAP glutamic acid decarboxylase GAD67


id: cord-257167-rz4r5sj7
type: CELL_LINE
entity:
  rat and mouse Yuriko Furukawa 1 2 Nahoko Kasai 1 Akiyoshi Shimada 1 Keiichi Torimitsu 1 2 Kunihiko Obata 3 Yuchio Yanagawa 2 4 Tadaharu Tsumoto 2 3 1 NTT Basic Research Laboratories Kanagawa Japan 2


id: cord-257167-rz4r5sj7
type: PROTEIN
entity:
  Ishikawa 1 Jun Shiota 1 Hiroyuki Tsutsumishita 1 Hiroyuki Sakagami 2 Masaaki Tsuda 1 Akiko Tabuchi 1 1 Dept Biol Chem Fac Pharm Sci Univ Toyama Toyama Japan 2


id: cord-257167-rz4r5sj7
type: DNA
entity:
  N terminal Otone Endo 1 2 Masaaki Mizuno 3 Yasukazu Kajita 2 Jun Yoshida 2 1 Department of Neurosurgery JA Kainan Hospital Aichi Japan 2 Department of Neurosurgery Nagoya University Nagoya Japan 3 Department of Molecular Neurosurgery Nagoya University Nagoya Japan Primate


id: cord-257167-rz4r5sj7
type: PROTEIN
entity:
  Yuka Okazaki 1 2 Arman Abrahamyan 3 Catherine Stevens 3 Andreas A Ioannides 1 2 1 Brain Science Institute


id: cord-257167-rz4r5sj7
type: PROTEIN
entity:
  Pax6 mutant mouse limbic system Hasumi Haba 1 Tadashi Nomura 1 Yoshinobu Hara 1 2 Noriko Osumi 1 2 1 Div Dev Neurosci CTAAR Tohoku Univ Sch Med Sendai Japan 2 CREST JST Japan Core


id: cord-257167-rz4r5sj7
type: DNA
entity:
  electroconvulsive shock Yoshifumi Ueta 1 Ryo Yamamoto 1 Shigeki Sugiura 2 Kaoru Inokuchi 3 Nobuo Kato 1 1 Dept Integrat Brain Sci Grad Sch Med Kyoto Univ 2 Nara Med Univ 3 Mitsubishi Kagaku Inst Life Sci


id: cord-257167-rz4r5sj7
type: CELL_LINE
entity:
  human cerebral hemisphere Taiko Kitamura Jinzo Yamada Department of Anatomy Tokyo Medical University Tokyo Japan


id: cord-257167-rz4r5sj7
type: DNA
entity:
  SNPs Keiko Yasumatsu 1 Sachiko Saito 2 Yuko Murata 3 Ding Ming 4 Tatsu Kobayakawa 5 Robert F Margolskee 4 Yuzo Ninomiya 1 1 Sect Oral Neurosci Grad Sch Dent Sci Kyushu Univ Fukuoka Japan 2


id: cord-257167-rz4r5sj7
type: PROTEIN
entity:
  ryanodine receptor type3 knockout mouse Suzuko Ohsako 1 Koichi Tanda 1 2 Nobuyuki Yamasaki 1 Keiko Toyama 1 Hiroshi Takeshima 3


id: cord-257167-rz4r5sj7
type: PROTEIN
entity:
  AMPA kainate glutamate receptors Tadahiro Katayama 1 Eiichi Jodo 1 Yoshiaki Suzuki 2 Ken yo Hoshino 2 Yukihiko Kayama 1 1


id: cord-257167-rz4r5sj7
type: CL
entity:
  stem and spinal myoclonus Cortical myoclonus is short in duration 50 ms By back averaging EEG or MEG time locked to spontaneous myoclonus a cortical activity is demonstrated in the corresponding area of the contralateral primary motor cortex immediately preceding the myoclonus by 20 ms for hand It is mediated by fact conducting corticospinal pathway Cortical myoclonus is often stimulus sensitive cortical reflex myoclonus showing extremely enhanced cortical responses to somatosensory or visual stimulus and enhanced longloop transcortical reflexes These findings together with transcranial magnetic stimulation suggest increased excitability of sensorimotor cortex in cortical myoclonus Mark Hallett NINDS Bethesda MD USA There have been many recent advances in the understanding of the physiology of focal dystonia Three main avenues of research have shown abnormalities in cortical inhibition sensory processing including sensorimotor integration and plasticity This lecture will emphasize the abnormal inhibition Abnormal inhibition appears to be the most direct cause of unwanted muscle contractions that make up both the involuntary spasms and the overflow movements also seen in this condition A loss of inhibition is seen in spinal and brainstem reflexes but these changes are likely secondary to cortical abnormalities Cortical inhibition is also diminished as demonstrated most clearly with transcranial magnetic stimulation GABA content may be decreased as shown with magnetic resonance spectroscopy A particular type of defective inhibition is surround inhibition the inhibition that normally operates to sharpen fine skilled movements Studies are now in progress to determine the synaptic


id: cord-257167-rz4r5sj7
type: GGP
entity:
  cytochrome C release and caspase activation induced by 6 OHDA ROS and p quinone SY3 2 07 6 Environmental factors


id: cord-257167-rz4r5sj7
type: GGP
entity:
  Hiromichi Sato 2 1 Grad Sch Front Biosci Osaka Univ Osaka Japan 2 Grad Sch Med Osaka Univ Japan 3 Grad Sch Eng Sci


id: cord-257167-rz4r5sj7
type: GGP
entity:
  Hiromichi Sato 1 1 Grad Sch Med Osaka Univ Japan 2 Med Sch Osaka Univ Japan 3 Grad Sch Front Biosci Osaka Univ Osaka Japan Spatial summation property of


id: cord-257167-rz4r5sj7
type: GGP
entity:
  Hiromichi Sato 2 3 1 Med Sch Osaka Univ 2 Grad Sch Med Osaka Univ 3 Grad Sch Front Biosci Osaka Univ Osaka Japan In the primary visual cortex


id: cord-257167-rz4r5sj7
type: GGP
entity:
  Yuko Sassa 1 2 Motoaki Sugiura 3 Hyeonjeong Jeong 1 4 Keisuke Wakusawa 1 5 Kaoru Horie 6 Shigeru Sato 6 Ryuta Kawashima 1 1


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Toshihisa Ohtsuka Department of Clinical and Molecular Pathology Faculty of Medicine Graduate School of Medicine University of Toyama


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Hideki Enomoto Laboratory for Neuronal Differentiation and Regeneration RIKEN Center for Developmental Biology Kobe


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Nature Hiroyuki Kato Center for Clinical Medicine and Research International University of Health and Welfare Nasushiobara


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Keiichiro Susuki Department of Neurology Dokkyo University School of Medicine Tochigi Japan Gangliosides


id: cord-257167-rz4r5sj7
type: ORG
entity:
  TrkA Kozo Kaibuchi Department of Cell Pharmacology Nagoya University Graduate School of Medicine Japan Neurons


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Hiroshi Toyoda Department of Cerebral Research National Institute for Physiological Sciences Okazaki Japan


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Neurosurgery Jichi Medical University Tochigi Japan 2 Department of Neurosurgery Tokyo Metropolitan Police Hospital Tokyo Mechanism


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Tamami Fukushi Research Institute of Science and Technology for Society RISTEX Japan Science and Technology Agency


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Tokyo Japan 2 Department of Applied System Neuroscience Nihon University Graduate School of Medical Science Tokyo


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Yoshiki Kaneoke Department of Integrative Physiology National Institute for Physiological Sciences Magnetoencephalography


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Setsuro Ibayashi Hiroaki Ooboshi Department of Medicine and Clinical Science Graduate School of Medical Sciences Kyushu University Fukuoka


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Toshiharu Iwasaki Noriyuki Koibuchi Department of Integrative Physiology Gunma University Graduate School of Medicine


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Singapore Singapore 2 Department of Neurophysiology Tokushima Bunri University Kagawa Japan 3 Department of Pharmacology Nippon Medical School Tokyo Japan Postsynaptic P2Y


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Komori Shigeki Moriguchi Department of Pharmacology Graduate School of Pharmaceutical Sciences Tohoku


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Psychology and Center for Neural Science New York University USA 2 Laboratory for Cognitive Brain Mapping RIKEN Brain Science Institute


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Takao Hirai Hiroaki Nishio Department of Molecular Pharmacology Faculty of Pharmacy and Pharmaceutical Sciences Fukuyama University


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Senzaki Takashi Shiga University of Tsukuba Graduate School of Comprehensive Human Sciences Tsukuba Japan Recent


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Mitsuko Sato Masako Miwa Mitsunobu Yoshii Division of Psychobiology Tokyo Institute of Psychiatry Tokyo


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Human Nutrition Notre Dame Seishin University Okayama Japan 2 Department of Curriculum Teaching and Memory Naruto University of Education


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Institute of Neuroscience Chinese Academy of Sciences China 2 Institute of Biophysics Chinese Academy of Sciences


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Hironobu Sakaguchi Department of Physiology and Biological Information Dokkyo University School of Medicine Japan Avian


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Yoshinori Izaki Tatsuo Akema Department of Physiology St Marianna University School of Medicine Japan


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Radiology and Nuclear Medicine Institute of Development Aging and Cancer Tohoku University


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Cognition and Action Group National Institute of Advanced Industrial Science and Technology AIST Ibaraki Japan 2 Department of Biomolecular Science Faculty of Science


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Tohru Kurotani Komatsu Yukio Department of Visual Neuroscience Research Institute of Environmental Medicine


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Kazuyoshi Kawa Department of Neurophysiology Tohoku University Graduate School of Medicine Sendai Japan Using


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Yasuyuki Ishikawa Sadao Shiosaka Division of Structural Cellular Biology Nara Institute of Science and Technology


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Kiyohisa Natsume Department of Brain Science and Engineering Kyushu Institute of Technology Kitakyushu


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Kyungpook National University School of Dentistry South Korea 2 Kyungpook National University School of Medicine


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Complexity Science and Engineering University of Tokyo Chiba Japan 4 Brain Science Institute RIKEN Saitama


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Masao Norita Department of Sensory and Integrative Medicine Niigata University Graduate School of Medical


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Bioengineering Nagaoka University of Technology Nagaoka Japan 2 Department of Clinical Pharmacy and Pharmacology Kagoshima University


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Kaizo Wen Jie Song Department of Electrical Electronic and Information Engineering Graduate School of Engineering


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Radiology National Hospital for Geriatric Medicine Obu Japan 2 Department of Neurosurgery National Hospital for Geriatric Medicine Obu Japan


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Dental University Tokyo Japan 2 Laboratory of Symbolic Cognitive Development RIKEN Brain Science Institute Wako Japan


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Neurophysiology Brain Research Institute Niigata University 2 Department Plastic Surgery School of Medicine Niigata University


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Anatomy Shinshu University School of Medicine Matsumoto Japan 2 Department of Pediatrics Shinshu University School of Medicine Matsumoto Japan


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Physiology School of Medicine University of Occupational and Environmental Health Kitakyushu Japan 2 Laboratory of Physiology Department of Marine Bioscience Ocean Research Institute University of Tokyo Japan 3 Department of Integrative Physiology University of Fukui


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Physiology School of Medicine University of Occupational and Environmental Health Kitakyushu Japan 2


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Physiology School of Medicine University of Occupational and Environmental Health Kitakyushu


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of physiology School of medicine University of Occupational and Environmental Health Japan 3 Department of physiology Jichi Medical School


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Suguru Kawato Department of Biophysics and Life Sciences Graduate School of Arts and Sciences University


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Physiology School of Medicine University of Occupational and Environmental Health Kitakyushu Japan 2 Peptide Institute


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Pharmacology 3 Department of Immunology School of Medicine University of Occupational and Environmental


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Anesthesiology and Critical Care Medicine Jichi Medical University Graduate School Tochigi Japan 2 Department of Physiology Jichi Medical University Tochigi Japan


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Curriculum Teaching and Memory Naruto University of Education Tokushima Japan 2 Department of Human Nutrition Notre Dame Seishin University Okayama Japan 3 Department of Physiology Saitama Medical School Saitama


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of pharmacology University of Kyoto Kyoto Japan 2 Department of Physiology University of Kurume


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Suzuki Hiroyoshi Miyakawa Laboratory of Cellular Neurobiology Tokyo University of Pharmacy and Life Science


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Otolaryngology Chiba University Chiba Japan 2 Department of Otolaryngology Kimitsu Central Hospital Chiba


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Integrative Neurophysiology Chiba University Chiba Japan 4 Section for Human Neurophysiology Research Center


id: cord-257167-rz4r5sj7
type: FAC
entity:
  Tatsuo Sato Iwao Fukui Harunori Ohmori Department of Physiology Faculty of Medicine Kyoto University Interaural


id: cord-257167-rz4r5sj7
type: ORG
entity:
  COE Program Iwate University Morioka Japan 2 Department of Welfare Engineering Faculty of Engineering Iwate University Morioka Japan


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Japan 5 Special Division for Human Life Technology National Institute of Advanced Industrial Science and Technology


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Pharmacology Graduate School of Medicine University of Tokyo Japan 3 Department of Anatomy Fukushima Medical University


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Applied Biology Kyoto Institute of Technology Kyoto Japan 2 Division of Bioscience Graduate School of Science and Technology Kobe


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Neuroscience Hyogo College of Medicine Hyogo Japan 2 Department of Otorhinolaryngology Hyogo College of Medicine Hyogo Japan


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Pharmacology Nippon Medical School Tokyo Japan 2 Pharmaceutical Research Center Kyowa Hakko Kogyo


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Katafuchi Megumu Yoshimura Department of Integrative Physiology Graduate School of Medical Sciences Kyushu University


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Narihito Iwashita Natsu Koyama Department of Physiology Shiga University of Medical Science Otsu Japan


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Miyagi University of Education Sendai Japan 4 Department of Pediatrics School of Medicine Tohoku University


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Anatomy Shiga University of Medical Science Otsu Japan 2 Fujita Health University School of Medicine Japan


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Graduated School of Integrated Sience Yokohama City University Yokohama Japan 2 Department of Psychology Hunter College City University of New York


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Japan 2 Department of Brain Science Graduate School of Medicine and Dentistry Okayama University Japan Disrupted


id: cord-257167-rz4r5sj7
type: ORG
entity:
  BSI Wako Japan 2 Department of Biophysics Kyoto University Kyoto Japan 3 Lab for Developmental Neurobiology


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Graduate School of Life Science and Systems Engineering Kyushu Institute of Technology Fukuoka Japan


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Okazaki Institute for Integrative Bioscience National Institute for Physiological Sciences Okazaki Japan Schizophrenia


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Mental Disorder Research National Institute of Neuroscience National Center of Neurology and Psychiatry Kodaira


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Neuropsychiatry School of Medicine Keio University 2 Department of Electrical and Electronics Engineering Sophia University


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Toshio Aoyagi 2 1 Department of Physics Kyoto University Kyoto Japan 2 Graduate School of Informatics Kyoto University Kyoto Japan


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Applied Biology Kyoto Institute of Technology Kyoto Japan 2 Department of Polymer Science and Engineering Kyoto Institute of Technology Kyoto


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Tokyo Japan 2 Department of Molecular Neurobiology Brain Research Institute Niigata University Niigata


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Developmental Biology Osaka University Graduate School of Dentistry Osaka Japan 2 First Department of Oral and Maxillofacial Surgery Osaka University Graduate School of Dentistry Osaka Japan Postsynaptic


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Rikako Kato Department of Developmental Physiology National Institute for Physiological Sciences Okazaki Japan Saccadic


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Dental University Tokyo Japan 2 Department of Physiology Ibaraki Prefectural University of Health Sciences Ibaraki


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Japan Science and Technology Agency JST Research Institute of Science and Technology for Society RISTEX


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Department of Physical Therapy University of Aino Osaka Japan 2 Institute for Frontier Medical Science Kyoto University Kyoto Japan Regeneration


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Anatomy Division of Microscopic and Developmental Anatomy Kurume University School of Medicine Kurume Japan Stimulation


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Iinuma Kenzo Hirose Department of Cell Physiology Nagoya University Graduate School of Medicine Nagoya Japan Glutamate


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Institute for Biomedical Engineering Consolidated Research Institute for Advanced Science and Medical Care Waseda University


id: cord-257167-rz4r5sj7
type: ORG
entity:
  Japan 2 Department of Medical System Engineering Faculty of Engineering Chiba University Chiba Japan


id: cord-257167-rz4r5sj7
type: CHEMICAL
entity:
  coconut juice N Radenahmad P Subhadhirasakul PSU Thailand Young coconut juice YCJ Cocos nucifera Arecaceae


id: cord-257167-rz4r5sj7
type: CHEMICAL
entity:
  N terminal BAR the central clathrin AP 2 binding and the C terminal SH3 domains and the variant specific two novel insertions A and B Insert A


id: cord-257167-rz4r5sj7
type: DISEASE
entity:
  cognitive functions Natsue Yoshimura 1 Daisuke Horiuchi 1 Tomoyuki Miyashita 2 Minoru Saitoe 2 Hitoshi Okazawa 1 1 Department of Neuropathology Medical Research Institute Tokyo Medical and Dental University Tokyo Japan 2 Tokyo Metropolitan Institute


# bib

id: cord-000933-nn9gj0z1
author: Krzyzaniak, Magdalena Anna
date: 2013-04-11
txt: ./txt/cord-000933-nn9gj0z1.txt
cache: ./cache/cord-000933-nn9gj0z1.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3623752/
doi: 10.1371/journal.ppat.1003309
summary:
  To study the entry process in human tissue culture cells (HeLa, A549), we used fluorescence microscopy and developed quantitative, FACS-based assays to follow virus binding to cells, endocytosis, intracellular trafficking, membrane fusion, and infection. To determine the pathway of RSV entry into HeLa and A549 cells, we developed quantitative fluorescence-activated cell sorting (FACS) assays and complemented them with confocal microscopy to monitor cell binding of RSV, endocytosis, fusion, and infection. Since we did not detect free capsids that would stain only for N or P (data not shown), we used the presence of the capsid antigens to distinguish between intact RSVs and VLPs. When purified virus preparations were incubated with HeLa cells at 4uC, immunoblotting after SDS-PAGE showed that more than half of the input N and P associated with the cells indicating that RSV binding in the cold was efficient (Fig. 1C) .

title:
  Host Cell Entry of Respiratory Syncytial Virus Involves Macropinocytosis Followed by Proteolytic Activation of the F Protein

abstract:
  Respiratory Syncytial Virus (RSV) is a highly pathogenic member of the Paramyxoviridae that causes severe respiratory tract infections. Reports in the literature have indicated that to infect cells the incoming viruses either fuse their envelope directly with the plasma membrane or exploit clathrin-mediated endocytosis. To study the entry process in human tissue culture cells (HeLa, A549), we used fluorescence microscopy and developed quantitative, FACS-based assays to follow virus binding to cells, endocytosis, intracellular trafficking, membrane fusion, and infection. A variety of perturbants were employed to characterize the cellular processes involved. We found that immediately after binding to cells RSV activated a signaling cascade involving the EGF receptor, Cdc42, PAK1, and downstream effectors. This led to a series of dramatic actin rearrangements; the cells rounded up, plasma membrane blebs were formed, and there was a significant increase in fluid uptake. If these effects were inhibited using compounds targeting Na(+)/H(+) exchangers, myosin II, PAK1, and other factors, no infection was observed. The RSV was rapidly and efficiently internalized by an actin-dependent process that had all hallmarks of macropinocytosis. Rather than fusing with the plasma membrane, the viruses thus entered Rab5-positive, fluid-filled macropinosomes, and fused with the membranes of these on the average 50 min after internalization. Rab5 was required for infection. To find an explanation for the endocytosis requirement, which is unusual among paramyxoviruses, we analyzed the fusion protein, F, and could show that, although already cleaved by a furin family protease once, it underwent a second, critical proteolytic cleavage after internalization. This cleavage by a furin-like protease removed a small peptide from the F1 subunits, and made the virus infectious.


id: cord-001123-n2e4s7bu
author: Lin, Yue-Zhi
date: 2013-11-22
txt: ./txt/cord-001123-n2e4s7bu.txt
cache: ./cache/cord-001123-n2e4s7bu.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3838338/
doi: 10.1371/journal.pone.0079299
summary:
  title: The Soluble Form of the EIAV Receptor Encoded by an Alternative Splicing Variant Inhibits EIAV Infection of Target Cells In addition to the previously described membrane-associated form of ELR1, two other major alternative splicing variant mRNAs were identified in equine monocyte-derived macrophages (eMDMs). This result strongly indicated that overexpressed recombinant sELR1 was able to inhibit EIAV infection in vitro, probably by competing with the membrane-associated receptor for the binding of virions on target cells. Thus, the soluble form of ELR1, tagged with HA, was overexpressed in FED cells, a major target of EIAV in vitro, by transfection with the sELR1 expression vector pcDNA3.1-ELR1-IN. The regulated expression levels of both the soluble and membrane-associated forms of ELR1 by EIAV reveal the possible role of sELR1 in the interaction between virus and host.

title:
  The Soluble Form of the EIAV Receptor Encoded by an Alternative Splicing Variant Inhibits EIAV Infection of Target Cells

abstract:
  Equine lentivirus receptor 1 (ELR1) has been identified as the sole receptor for equine infectious anemia virus (EIAV) and is a member of the tumor necrosis factor receptor (TNFR) superfamily. In addition to the previously described membrane-associated form of ELR1, two other major alternative splicing variant mRNAs were identified in equine monocyte-derived macrophages (eMDMs). One major spliced species (ELR1-IN) contained an insertion of 153 nt, which resulted in a premature stop codon situated 561 nt upstream of the predicted membrane spanning domain. The other major species (ELR1-DE) has a deletion of 109 nt that causes a shift of the open reading frame and generates a stop codon 312 nt downstream. Because ELR1-DE presumably encodes a peptide of a mere 23 residues, only ELR1-IN was further analyzed. The expression of a soluble form of ELR1 (sELR1) by ELR1-IN was confirmed by Western blot and immunofluorescence analyses. Similar to ELR1, the transcription level of ELR1-IN varied among individual horses and at different time points in the same individuals. The ratio of ELR1-IN mRNA species to ELR1 mRNA was approximately 1∶2.5. Pre-incubation of the recombinant sELR1 with EIAV significantly inhibited EIAV infection in equine macrophages, the primary in vivo target cell of the virus. Fetal equine dermal (FED) cells are susceptible to EIAV in vitro, and the replication of EIAV in FED cells transiently transfected with ELR1-IN was markedly reduced when compared with replication in cells transfected with the empty vector. Finally, the expression levels of both forms of the EIAV receptor were significantly regulated by infection with this virus. Taken together, our data indicate that sELR1 acts as a secreted cellular factor that inhibits EIAV infection in host cells.


id: cord-001270-l8aa9cl3
title: Antiviral restriction factor transgenesis in the domestic cat
author: Wongsrikeao, Pimprapar
date: 2011-09-11
txt: ./txt/cord-001270-l8aa9cl3.txt
cache: ./cache/cord-001270-l8aa9cl3.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4006694/
doi: 10.1038/nmeth.1703
summary:
  In contrast to primates, feline species lack antiviral TRIM5α genes 11 but have potently restrictive APOBEC3 proteins 9, 10 , which sets up intriguing possibilities for testing such genes at the whole-animal level, for conferring gene-based immunity with them or engineered variants 12, 13 , and potentially for HIV-1 disease model development 10 . In experiments summarized in Supplementary Table 1 , we subjected 195 in vitro-matured grade I and II domestic cat oocytes to perivitelline space microinjection (PVSMI) with lentiviral vector TSinG 5 ; we performed injection 10-12 h before or 10-12 h after in vitro fertilization (IVF) (Supplementary Fig. 1 ). We consistently observed embryo-pervasive, abundant expression of both proteins encoded by dual gene vectors in cat blastocysts when we injected lentiviral vector before IVF ( Fig. 1b and Supplementary Table 2 ).

abstract:
  Studies of the domestic cat have contributed to many scientific advances, including the present understanding of the mammalian cerebral cortex. A practical capability for cat transgenesis is needed to realize the distinctive potential of research on this neurobehaviorally complex, accessible species for advancing human and feline health. For example, humans and cats are afflicted with pandemic AIDS lentiviruses that are susceptible to species-specific restriction factors. Here we introduced genes encoding such a factor, rhesus macaque TRIMCyp, and eGFP, into the cat germline. The method establishes gamete-targeted transgenesis for the first time in a carnivore. We observed uniformly transgenic outcomes, widespread expression, no mosaicism and no F1 silencing. TRIMCyp transgenic cat lymphocytes resisted feline immunodeficiency virus replication. This capability to experimentally manipulate the genome of an AIDS-susceptible species can be used to test the potential of restriction factors for HIV gene therapy and to build models of other infectious and noninfectious diseases. SUPPLEMENTARY INFORMATION: The online version of this article (doi:10.1038/nmeth.1703) contains supplementary material, which is available to authorized users.


id: cord-001365-6u80p5sj
title: A Novel MVA Vectored Chikungunya Virus Vaccine Elicits Protective Immunity in Mice
author: Weger-Lucarelli, James
date: 2014-07-24
txt: ./txt/cord-001365-6u80p5sj.txt
cache: ./cache/cord-001365-6u80p5sj.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4109897/
doi: 10.1371/journal.pntd.0002970
summary:
  Despite the presence of high levels of neutralizing antibodies elicited by most of the VACV vectored VEEV vaccine candidates, they were ineffective in providing protection against airborne infection, suggesting they were unable to elicit a sufficient T cell mediated immune response, which has been shown to be critical for protection against lethal VEEV encephalitis [34, 35] . Most importantly, depletion of CD4 + T cells in vaccinated mice resulted in loss of protection, with 100% succumbing to infection upon challenge with wild-type CHIKV, indicating an indispensable role of MVA-CHIK immune CD4+ T cells in protection. Expression of E2 was observed in the supernatant of CHIKV infected cells, but there Prior to boost (where applicable) and challenge, mice were bled and serum was monitored for both total Ig(G+M) and neutralizing activity by TCID 50 was no detection of E2 expression in MVA-CHIK, suggesting that the protein was not being secreted.

abstract:
  BACKGROUND: Chikungunya virus (CHIKV) is a re-emerging arbovirus associated with febrile illness often accompanied by rash and arthralgia that may persist for several years. Outbreaks are associated with high morbidity and create a public health challenge for countries affected. Recent outbreaks have occurred in both Europe and the Americas, suggesting CHIKV may continue to spread. Despite the sustained threat of the virus, there is no approved vaccine or antiviral therapy against CHIKV. Therefore, it is critical to develop a vaccine that is both well tolerated and highly protective. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we describe the construction and characterization of a modified Vaccinia virus Ankara (MVA) virus expressing CHIKV E3 and E2 proteins (MVA-CHIK) that protected several mouse models from challenge with CHIKV. In particular, BALB/c mice were completely protected against viremia upon challenge with CHIKV after two doses of MVA-CHIK. Additionally, A129 mice (deficient in IFNα/β) were protected from viremia, footpad swelling, and mortality. While high anti-virus antibodies were elicited, low or undetectable levels of neutralizing antibodies were produced in both mouse models. However, passive transfer of MVA-CHIK immune serum to naïve mice did not protect against mortality, suggesting that antibodies may not be the main effectors of protection afforded by MVA-CHIK. Furthermore, depletion of CD4(+), but not CD8(+) T-cells from vaccinated mice resulted in 100% mortality, implicating the indispensable role of CD4(+) T-cells in the protection afforded by MVA-CHIK. CONCLUSIONS/SIGNIFICANCE: The results presented herein demonstrate the potential of MVA to effectively express CHIKV E3-E2 proteins and generate protective immune responses. Our findings challenge the assumption that only neutralizing antibodies are effective in providing protection against CHIKV, and provides a framework for the development of novel, more effective vaccine strategies to combat CHIKV.


id: cord-002076-7t4d4vvo
author: Li, Yongfeng
date: 2016-05-06
txt: ./txt/cord-002076-7t4d4vvo.txt
cache: ./cache/cord-002076-7t4d4vvo.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4885082/
doi: 10.3390/v8050127
summary:
  Commonly used tests based on wild-type viruses, such as immunostaining, cannot meet the demands for rapid detection of viral replication, high-throughput screening for antivirals, as well as for tracking viral proteins or virus transport in real time. This article reviews the applications of RCREVs in diagnostic and molecular virology, including rapid neutralization tests, high-throughput screening systems, identification of viral receptors and virus-host interactions, dynamics of viral infections in vitro and in vivo, vaccination approaches and others. Replicating-competent reporter-expressing viruses (RCREVs) are one type of artificially modified viruses that not only retain the viral genetic characteristics but also possess the new properties of the reporter genes, which represent a useful tool for quantitative analysis of viral replication and tracking viral protein transport in both living cells and animals.

title:
  Applications of Replicating-Competent Reporter-Expressing Viruses in Diagnostic and Molecular Virology

abstract:
  Commonly used tests based on wild-type viruses, such as immunostaining, cannot meet the demands for rapid detection of viral replication, high-throughput screening for antivirals, as well as for tracking viral proteins or virus transport in real time. Notably, the development of replicating-competent reporter-expressing viruses (RCREVs) has provided an excellent option to detect directly viral replication without the use of secondary labeling, which represents a significant advance in virology. This article reviews the applications of RCREVs in diagnostic and molecular virology, including rapid neutralization tests, high-throughput screening systems, identification of viral receptors and virus-host interactions, dynamics of viral infections in vitro and in vivo, vaccination approaches and others. However, there remain various challenges associated with RCREVs, including pathogenicity alterations due to the insertion of a reporter gene, instability or loss of the reporter gene expression, or attenuation of reporter signals in vivo. Despite all these limitations, RCREVs have become powerful tools for both basic and applied virology with the development of new technologies for generating RCREVs, the inventions of novel reporters and the better understanding of regulation of viral replication.


id: cord-002320-m99amd4y
author: Mathur, Kalika
date: 2016-11-30
txt: ./txt/cord-002320-m99amd4y.txt
cache: ./cache/cord-002320-m99amd4y.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5128919/
doi: 10.1038/srep38065
summary:
  Systematic analysis of all CHIKV proteins using a Sf21 RNAi sensor cell line based assay revealed that non-structural proteins nsP2 and nsP3 exhibited RNAi suppressor activity. Using cell lysate of the transfected cell expressing nsP2 and nsP3, we evaluated their ability to bind double stranded RNA by using [γ 32P] labeled shRNA of GFP and running the bound complex in a 6% polyacrylamide gel (Fig. 3e) . Having confirmed that both nsP2 and nsP3 exhibit RNAi suppressor activities, efforts were taken to characterise the proteins better and to identify those specific domains that were and NS4B (dengue virus) were taken as positive controls and empty vector was used as negative control. Taken together, the current study has identified CHIKV proteins nsP2 and nsP3 to exhibit RNAi suppressor activity in the in-vitro system that was further demonstrated in its natural hosts, namely, an Aedes and mammalian cell line.

title:
  Analysis of chikungunya virus proteins reveals that non-structural proteins nsP2 and nsP3 exhibit RNA interference (RNAi) suppressor activity

abstract:
  RNAi pathway is an antiviral defence mechanism employed by insects that result in degradation of viral RNA thereby curbing infection. Several viruses including flaviviruses encode viral suppressors of RNAi (VSRs) to counteract the antiviral RNAi pathway. Till date, no VSR has been reported in alphaviruses. The present study was undertaken to evaluate chikungunya virus (CHIKV) proteins for RNAi suppressor activity. We systematically analyzed all nine CHIKV proteins for RNAi suppressor activity using Sf21 RNAi sensor cell line based assay. Two non-structural proteins, namely, nsP2 and nsP3 were found to exhibit RNAi suppressor activity. We further validated the findings in natural hosts, namely in Aedes and in mammalian cell lines and further through EMSA and Agrobacterium infiltration in GFP silenced transgenic tobacco plants. Domains responsible for maximum RNAi suppressor activity were also identified within these proteins. RNA binding motifs in these domains were identified and their participation in RNAi suppression evaluated using site directed mutagenesis. Sequence alignment of these motifs across all species of known alphaviruses revealed conservation of these motifs emphasizing on a similar role of action in other species of alphaviruses as well. Further validation of RNAi suppressor activity of these proteins awaits establishment of specific virus infection models.


id: cord-003705-ekhj8ae8
author: Azkanaz, Maria
date: 2019-06-14
txt: ./txt/cord-003705-ekhj8ae8.txt
cache: ./cache/cord-003705-ekhj8ae8.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6570483/
doi: 10.7554/elife.45205
summary:
  To verify the HS-induced accumulation of PcG proteins in the nucleolus, we isolated nucleoli from heat shocked and untreated GFP-CBX8 expressing K562 cells ( Figure 2A ). Whereas initially considered as heat-induced damage, several lines of independent observations have suggested that this might rather reflect a regulated, HSP-dependent process in which the nucleolus serves as a temporal storage site for unfolded proteins during proteotoxic stress (Nollen et al., 2001; Ohtsuka et al., 1986; Welch and Feramisco, 1984) . Intra-nucleolar levels of H3K27me3 and H2AK119ub were not increased in heat shocked cells versus untreated cells, suggesting that PcG proteins are not involved in Polycomb-mediated silencing of nucleolar chromatin ( Figure 3E ). Independent LC-MS/MS analysis with K562 GFP-CBX8 cells confirmed these findings, implying that HS-induced nucleolar accumulation of various chromatin regulators, protein chaperones and proteasomal subunits is a conserved biological phenomenon (Figure 3-figure supplement 4A-E, Supplementary file 3).

title:
  Protein quality control in the nucleolus safeguards recovery of epigenetic regulators after heat shock

abstract:
  Maintenance of epigenetic modifiers is of utmost importance to preserve the epigenome and consequently appropriate cellular functioning. Here, we analyzed Polycomb group protein (PcG) complex integrity in response to heat shock (HS). Upon HS, various Polycomb Repressive Complex (PRC)1 and PRC2 subunits, including CBX proteins, but also other chromatin regulators, are found to accumulate in the nucleolus. In parallel, binding of PRC1/2 to target genes is strongly reduced, coinciding with a dramatic loss of H2AK119ub and H3K27me3 marks. Nucleolar-accumulated CBX proteins are immobile, but remarkably both CBX protein accumulation and loss of PRC1/2 epigenetic marks are reversible. This post-heat shock recovery of pan-nuclear CBX protein localization and reinstallation of epigenetic marks is HSP70 dependent. Our findings demonstrate that the nucleolus is an essential protein quality control center, which is indispensable for recovery of epigenetic regulators and maintenance of the epigenome after heat shock.


id: cord-004534-jqm1hxps
title: Abstract
author: nan
date: 2009-06-09
txt: ./txt/cord-004534-jqm1hxps.txt
cache: ./cache/cord-004534-jqm1hxps.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7079852/
doi: 10.1007/s00249-009-0478-1
abstract: nan
summary:
  HIV-1 to efficiently complete a replication cycle has to integrate its genome into the host cellular DNA.After HIV-1 enters target cells,neosynthesized viral DNA forms along with other proteins the pre-integration complex (PIC).PICs are then transported into the nucleus where integration,catalyzed by the viral integrase,takes place.HIV-1 viral particles engineered to incorporate integrase fused to EGFP have proven effective to study PICs within nuclei of infected cells.In this study we report the live imaging analysis of nuclear PIC dynamics obtained by time-lapse microscopy.Intranuclear trajectories of IN-EGFP-labeled PIC were collected in three dimensions and examined by both mean squared displacement (MSD) and cage diameter (CD) analysis.In CD the maximum distances measured between two positions occupied by a PIC in a time window of 2 minutes were calculated while in our MSD analysis 5-minute long trajectory segments were considered.Remarkably,MSD revealed the presence of an underlying active transport mechanism.To test the possible role of actin filaments,PIC nuclear trafficking was analyzed in cells treated with latrunculin B (actin polymerization inhibitor).Preliminary results suggest that the disruption of actin function impairs the active nuclear movement of PICs. Second harmonic generation microscopy reveals sarcomere contractile dynamics of cardiomyocytes N.


id: cord-018771-xqlb74px
title: Bluttransfusion
author: Turinsky, Sebastian
date: 2013-04-04
txt: ./txt/cord-018771-xqlb74px.txt
cache: ./cache/cord-018771-xqlb74px.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7123737/
doi: 10.1007/978-3-642-34433-6_4
summary:
  Ein leukozytendepletiertes EK wird unmittelbar vor der Transfusion gewaschen und muss dann innerhalb von 6 h ohne weitere Lagerung transfundiert werden. Sowohl der zunehmende Mangel an Blutprodukten als auch diverse andere Umstände wie die Ablehnung durch den Patienten oder eine verzögerte Verfügbarkeit von Blutprodukten im Notfall machen es erforderlich, Alternativen zur Transfusion von Fremdblut zu kennen. B. bei schwerer Blutungsanämie, durch Beatmung mit einer FiO 2 = 1,0 eine Zunahme des physikalisch im Blut gelösten Sauerstoffs erreicht werden, die etwa dem Effekt der Transfusion von 1-2 EK entspricht. Bei schweren anaphylaktischen Transfusionsreaktionen, wie sie häufiger bei Patienten mit IgA-Mangel und der Ausbildung von Anti-IgA-Antikörpern auftreten können, besteht die Indikation zur Transfusion gewaschener Blutprodukte. Ist im Rahmen einer lebensbedrohlichen Situation eine Transfusion mit Rhesus-positivem Blut unvermeidlich, so kann die Bildung von Antikörpern durch eine Immunisierung gegen das Rhesusantigen verhindert werden (Gabe von Anti-D-Immunglobulin, z.

abstract:
  Bei einem 58-jährigen Patienten mit einem Hb-Wert von 6,5 g/dl, der nach operativer Ausschaltung eines Bauchaortenaneurysmas auf der Intensivstation unerwartet nachblutet, soll eine Bluttransfusion durchgeführt werden. Wenige Minuten nach der Anforderung aus der Blutbank treffen die bestellten Erythrozytenkonzentrate ein – es sind Erythrozytenkonzentrate der Blutgruppe A. Sie sind eindeutig auf den Namen des Patienten ausgezeichnet, die Identität des Patienten wird erneut überprüft. Der durchgeführte Bedside-Test zeigt aber die Blutgruppe B! Was muss der Intensivarzt nun tun?


id: cord-023026-2r84ndzv
title: Posters
author: nan
date: 2013-06-14
txt: ./txt/cord-023026-2r84ndzv.txt
cache: ./cache/cord-023026-2r84ndzv.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7165910/
doi: 10.1002/glia.22530
abstract: nan
summary:
  Thus, this work provides the basis to identify molecular pathways regulated by distinct niche/environmental signals and involved in the heterogeneity of adult OPCs. Multiple sclerosis (MS) is a chronic inflammatory and neurodegenerative demyelinating disease of the central nervous system (CNS) characterized by inflammation, which leads to formation of demyelinating areas due to loss of oligodendrocytes, astrogliosis and, finally, axonal degeneration. Taken together, these results demonstrate the important role of miR-200b in modulating the MAPK pathway via c-Jun which in turn affects different aspects of the inflammatory process accompanying microglia activation including cytokine response, NO production, phagocytosis and neuronal cell death. For this purpose, coronal cryostat free-floating sections from the brain of both adult transgenic mice and their corresponding wild-type (Wt) littermates, were processed for the study of astrocytes using GFAP immunohistochemistry and microglia using antibodies against Iba1 and several markers commonly related to the activated phenotype of these microglial cells, such as CD16/32 (Fc receptor), F4/80, CD11b, CD206, CD150 and MHC-II.


id: cord-023211-kt5gt26t
title: Poster Session Abstracts
author: nan
date: 2007-08-29
txt: ./txt/cord-023211-kt5gt26t.txt
cache: ./cache/cord-023211-kt5gt26t.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7167830/
doi: 10.1002/ppul.20700
abstract: nan
summary:
  Previous studies performed using fluorescence halide efflux measurements and short-circuit current voltage clamp have shown that treatment with PPARγ (peroxisome proliferator activated receptor gamma) agonists, such as pioglitazone and FLL (FMOC-L-leucine), resulted in an increased biosynthesis and trafficking of ∆F508-CFTR to the cell surface. Physiology, School of Medical Sciences, University of Bristol, Bristol, United Kingdom Recent progress in the development of small molecule correctors and potentiators capable of restoring CFTR function have increased the need for pre-clinical test models including cultured airway epithelial cells from human CF patients as well as CF mouse models. Clinical studies have linked increased sputum and peripheral blood neutrophil MPO activity with increased airflow obstruction in cystic fibrosis (CF) patients of the same age, gender, airway bacterial flora, and CFTR genotype. Because patients expressing low levels of normal CFTR mRNA (5-20%) have mild disease symptoms, these studies demonstrate that the incorporation of the ciliated cell-specific FOXJ1 promoter into gene therapy vectors may be useful for treatment of CF.


id: cord-102184-8u73adnk
title: Visual input into the Drosophila melanogaster mushroom body
author: Li, Jinzhi
date: 2020-05-02
txt: ./txt/cord-102184-8u73adnk.txt
cache: ./cache/cord-102184-8u73adnk.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1101/2020.02.07.935924
doi: 10.1101/2020.02.07.935924
summary:
  Here, we use a range of anatomical and genetic techniques to identify two novel types of mushroom body input neuron that connect visual processing centers — namely the lobula and the posterior lateral protocerebrum — to the dorsal accessory calyx of the mushroom body. Altogether, based on the results obtained using both the GRASP technique and the dye/photo-labeling technique, we conclude that LOPN is a true input 265 neuron of the a/bp Kenyon cells and that it conveys information from the lobula, a visual processing center, to the dorsal accessory calyx. To identify the neurons that project to the post-synaptic terminals formed by the PLPPNs in the posterior lateral protocerebrum, a poorly characterized visual processing center, we used the targeted photo-labeling technique described above ( Figure 8A ). Our results suggest that the LOPN and PLPPNs that we identified in our screen connect to the a/bp Kenyon cells and that, together, these two types of projection neuron represent a large fraction of the input neurons of the dorsal accessory calyx.

abstract:
  The ability to integrate input from different sensory systems is a fundamental property of many brains. Yet, the patterns of neuronal connectivity that underlie such multisensory integration remain poorly characterized. The Drosophila melanogaster mushroom body — an associative center required for the formation of olfactory and visual memories — is an ideal system to investigate how different sensory channels converge in higher-order brain centers. The neurons connecting the mushroom body to the olfactory system have been described in great detail, but input from other sensory systems remains poorly defined. Here, we use a range of anatomical and genetic techniques to identify two novel types of mushroom body input neuron that connect visual processing centers — namely the lobula and the posterior lateral protocerebrum — to the dorsal accessory calyx of the mushroom body. Together with previous work that described a pathway conveying visual information from the medulla to the ventral accessory calyx of the mushroom body (Vogt et al., 2016), our study defines a second, parallel pathway that is anatomically poised to convey information from the visual system to the dorsal accessory calyx. This connectivity pattern — the segregation of the visual information into two separate pathways — could be a fundamental feature of the neuronal architecture underlying multisensory integration in associative brain centers.


id: cord-103509-hynnba03
title: A self-assembled protein nanoparticle serving as a one-shot vaccine carrier
author: Wong, Ten-Tsao
date: 2020-09-18
txt: ./txt/cord-103509-hynnba03.txt
cache: ./cache/cord-103509-hynnba03.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1101/2020.09.16.299149
doi: 10.1101/2020.09.16.299149
summary:
  From these two clues, it was hypothesized that AH3-GFP fusion protein form stable protein complex through hydrophobic interaction mediated by N-terminal AH3 peptide. Using the same protocol, AH3-GFP was found to co-sedimented with the bacterial membrane (Fig. S3A) as well as the AH3-sfGFP-hM2e fusion protein but not a free GFP protein (Fig S3B) These results are consistent with our protein structure modeling that Arg11 serves as a main contact point for dimer stacking also it mediates the electrostatic interaction with mutated Glu13 (Fig. 3E ). These results suggest that the two point mutations of AH3 in I8L and K13E enable the formation of a stable, high antigenic protein complex that stimulates long lasting immune responses in a single immunization. The result suggests that although carrier protein AH3-sfGFP also has high antigenicity, it did not interfere with the immune response against the heterologous protein, hM2e (Fig. 5B, 5C) 

abstract:
  In this paper, we are exploring the role of an amphipathic helical peptide in mediating the self-assembly of a fusion protein into a protein nanoparticle and the application of the nanoparticle as a one-shot vaccine carrier. Out of several candidates, an amphipathic helical peptide derived from M2 protein of type A influenza virus is found to stimulate high antigenicity when fused to a fluorescent protein genetically. This fusion protein was found to form protein nanoparticle spontaneously when expressed and purified protein stimulates long-lasting antibody responses in single immunization. Through modeling peptide structure and nanoparticle assembly, we have improved this vaccine carrier in complex stability. The revised vaccine carrier is able to stimulate constant antibody titer to a heterologous antigen for at least six months in single immunization. The immune response against a heterologous antigen can be boosted further by additional immunization in spite of high immune responses to carrier protein.


id: cord-103868-iwpiti2h
title: The Ebola virus interferon antagonist VP24 undergoes active nucleocytoplasmic trafficking
author: Harrison, Angela R.
date: 2020-08-11
txt: ./txt/cord-103868-iwpiti2h.txt
cache: ./cache/cord-103868-iwpiti2h.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1101/2020.08.10.245563
doi: 10.1101/2020.08.10.245563
summary:
  Functions of the Ebola virus (EBOV) IFN antagonist VP24 include nucleocapsid assembly during cytoplasmic replication and inhibition of IFN-activated signalling by STAT1. Proteins of many viruses with cytoplasmic replication cycles similar to EBOV interact with the nuclear trafficking machinery, resulting in active nucleocytoplasmic shuttling important to immune evasion and other intranuclear functions. Thus, it appears that active In this study we have shown that EBOV VP24 undergoes active trafficking between the nucleus 275 and cytoplasm involving CRM1-dependent nuclear export via a NES at the VP24 C-terminus. Notably, the EBOV 287 matrix protein VP40 has also been reported to localise to the nucleus in infected and transfected 288 cells (16, 50); however, a direct role for active trafficking pathways to regulate localisation, 289 distinct from mechanisms such as diffusion or interaction with other host factors, has not been 290 defined.

abstract:
  Viral interferon (IFN) antagonist proteins mediate evasion of IFN-mediated innate immunity and are often multifunctional, having distinct roles in viral replication processes. Functions of the Ebola virus (EBOV) IFN antagonist VP24 include nucleocapsid assembly during cytoplasmic replication and inhibition of IFN-activated signalling by STAT1. For the latter, VP24 prevents STAT1 nuclear import via competitive binding to nuclear import receptors (karyopherins). Many viral proteins, including proteins from viruses with cytoplasmic replication cycles, interact with the trafficking machinery to undergo nucleocytoplasmic transport, with key roles in pathogenesis. Despite established karyopherin interaction, the nuclear trafficking profile of VP24 has not been investigated. We find that VP24 becomes strongly nuclear following overexpression of karyopherin or inhibition of nuclear export pathways. Molecular mapping indicates that cytoplasmic localisation of VP24 depends on a CRM1-dependent nuclear export sequence at the VP24 C-terminus. Nuclear export is not required for STAT1 antagonism, consistent with competitive karyopherin binding being the principal antagonistic mechanism while export mediates return of nuclear VP24 to the cytoplasm for replication functions. Thus, nuclear export of VP24 might provide novel targets for antiviral approaches. Importance Ebola virus (EBOV) is the causative agent of ongoing outbreaks of severe haemorrhagic fever with case-fatality rates between 40 and 60%. Proteins of many viruses with cytoplasmic replication cycles similar to EBOV interact with the nuclear trafficking machinery, resulting in active nucleocytoplasmic shuttling important to immune evasion and other intranuclear functions. However, exploitation of host trafficking machinery for nucleocytoplasmic transport by EBOV has not been directly examined. We find that the EBOV protein VP24 is actively trafficked between the nucleus and cytoplasm, and identify the specific pathways and sequences involved. The data indicate that nucleocytoplasmic trafficking is important for the multifunctional nature of VP24, which has critical roles in immune evasion and viral replication, identifying a new mechanism in infection by this highly lethal pathogen, and potential target for antivirals.


id: cord-253709-frauasts
author: Lan, Dongming
date: 2011-08-15
txt: ./txt/cord-253709-frauasts.txt
cache: ./cache/cord-253709-frauasts.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1016/j.ab.2011.04.034
doi: 10.1016/j.ab.2011.04.034
summary:
  title: An improved nonchromatographic method for the purification of recombinant proteins using elastin-like polypeptide-tagged proteases Abstract Proteins fused to the elastin-like polypeptide (ELP) tag can be selectively separated from crude cell extract without chromatography. Proteins fused to the elastin-like polypeptide (ELP) tag can be selectively separated from crude cell extract without chromatography. Among them, Meyer and Chilkoti reported a nonchromatographic method for purification of recombinant proteins using an elastin-like polypeptide (ELP) 2 tag [1] . This characteristic transition allows the recombinant ELP fusion protein to be isolated from the cell lysate by repeated steps of aggregation, centrifugation, and resolubilization without chromatography. Recombinant ELP fusion proteins at different expression levels can be efficiently recovered by the ITC method [4, 5] . ELP-3C protease was produced at a high level with an apparent molecular weight of approximately 58 kDa. To aggregate ELP-3C protease, solid NaCl was added into the cleared lysate to a final concentration of 2 M at room temperature to trigger the phase transition of ELP.

title:
  An improved nonchromatographic method for the purification of recombinant proteins using elastin-like polypeptide-tagged proteases

abstract:
  Abstract Proteins fused to the elastin-like polypeptide (ELP) tag can be selectively separated from crude cell extract without chromatography. To avoid the interference of the ELP tag on properties of the target protein, it is necessary to remove the ELP tag from target protein by protease digestion. Therefore, an additional chromatographic purification step is required to remove the proteases, and this is time- and labor-consuming. Here we demonstrate the utility of the ELP-tagged proteases for cleavage of ELP fusion proteins, allowing one-step removal of the cleaved ELP tag and ELP-tagged proteases without chromatography.


id: cord-257167-rz4r5sj7
title: Abstracts for the 29th Annual Meeting of the Japan Neuroscience Society (Neuroscience2006)
author: nan
date: 2006-12-31
txt: ./txt/cord-257167-rz4r5sj7.txt
cache: ./cache/cord-257167-rz4r5sj7.txt
extension: .txt
mime: text/plain
url: https://api.elsevier.com/content/article/pii/S016801020600085X
doi: 10.1016/j.neures.2006.04.004
abstract: nan
summary:
  SY1-3-11-3 SAD: A novel kinase implicated in phosphoproteome at the presynaptic active zone Toshihisa Ohtsuka Department of Clinical and Molecular Pathology, Faculty of Medicine/Graduate School of Medicine, University of Toyama, Toyama, Japan SAD is a serine/threonine kianse, which has been shown to regulate various neuronal functions during development, including clustering synaptic vesicles, maturation of synapses, and axon/dendrite polarization: these have recently been revealed by genetic studies in C. The results suggest that EAAT4 plays a major role in regulating the concentration of CF transmitters, possibly glutamate, in the route of its extrasynaptic diffusion, and determining the degree of CF-induced inhibition of GABA release from BCs depending on the regional difference of EAAT4 expression in postsynaptic PCs. Chitoshi Takayama 1 , Yoshiro Inoue 1 1 Department of Molecular Neuroanatomy, Hokkaido University School of Medicine, Sapporo, Japan GABA mediates inhibitory transmission in the adult central nervous system (CNS).


id: cord-258678-0atfsivf
title: A Universal Protein Tag for Delivery of SiRNA-Aptamer Chimeras
author: Liu, Hong Yan
date: 2013-11-07
txt: ./txt/cord-258678-0atfsivf.txt
cache: ./cache/cord-258678-0atfsivf.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/24196104/
doi: 10.1038/srep03129
summary:
  Here, we report the development of a small and simple protein tag that complements the therapeutic and targeting functionalities of chimera with two functional domains: a dsRNA binding domain (dsRBD) for siRNA docking and a pH-dependent polyhistidine to disrupt endosomal membrane. Therefore, it is of critical importance to design a delivery system that is simple for potential regulatory approval and mass production, universal for all siRNAaptamer chimera, neutral and siRNA-binding specific to ensure aptamer targeting, and small to avoid major alteration of chimera''s biodistribution profile. To assess their dsRNA binding activity, siRNA-aptamer chimera labeled with fluorophore FAM were incubated with the protein tags and probed with gel electrophoresis (1% agarose). To evaluate the targeting specificity of the aptamer block, PSMA-positive LNCaP cells and PSMA-negative PC3 cells were treated with complex of chimera and dsRBD-His 18 (chimera/protein tag molar ratio at 152, 100 nM chimera) in serum free medium for 2 hours, followed by incubation in complete medium for another 12 h.

abstract:
  siRNA-aptamer chimeras have emerged as one of the most promising approaches for targeted delivery of siRNA due to the modularity of their diblock RNA structure, relatively lower cost over other targeted delivery approaches, and, most importantly, the outstanding potential for clinical translation. However, additional challenges must be addressed for efficient RNA interference (RNAi), in particular, endosomal escape. Currently, vast majority of siRNA delivery vehicles are based on cationic materials, which form complexes with negatively charged siRNA. Unfortunately, these approaches complicate the formulations again by forming large complexes with heterogeneous sizes, unfavorable surface charges, colloidal instability, and poor targeting ligand orientation. Here, we report the development of a small and simple protein tag that complements the therapeutic and targeting functionalities of chimera with two functional domains: a dsRNA binding domain (dsRBD) for siRNA docking and a pH-dependent polyhistidine to disrupt endosomal membrane. The protein selectively tags along the siRNA block of individual chimera, rendering the overall size of the complex small, desirable for deep tissue penetration, and the aptamer block accessible for target recognition. More interestingly, we found that extending the c-terminal polyhistidine segment in the protein tag to 18 amino acids completely abolishes the RNA binding function of dsRBD.


id: cord-272018-txdc0c3j
title: Enhanced Autophagy Contributes to Reduced Viral Infection in Black Flying Fox Cells
author: Laing, Eric D.
date: 2019-03-14
url: https://doi.org/10.3390/v11030260
doi: 10.3390/v11030260
abstract:
  Bats are increasingly implicated as hosts of highly pathogenic viruses. The underlying virus–host interactions and cellular mechanisms that promote co-existence remain ill-defined, but physiological traits such as flight and longevity are proposed to drive these adaptations. Autophagy is a cellular homeostatic process that regulates ageing, metabolism, and intrinsic immune defense. We quantified basal and stimulated autophagic responses in black flying fox cells, and demonstrated that although black flying fox cells are susceptible to Australian bat lyssavirus (ABLV) infection, viral replication is dampened in these bat cells. Black flying fox cells tolerated prolonged ABLV infection with less cell death relative to comparable human cells, suggesting post-entry mechanisms interference with virus replication. An elevated basal autophagic level was observed and autophagy was induced in response to high virus doses. Pharmacological stimulation of the autophagy pathway reduced virus replication, indicating autophagy acts as an anti-viral mechanism. Enhancement of basal and virus-induced autophagy in bat cells connects related reports that long-lived species possess homeostatic processes that dampen oxidative stress and macromolecule damage. Exemplifying the potential that evolved cellular homeostatic adaptations like autophagy may secondarily act as anti-viral mechanisms, enabling bats to serve as natural hosts to an assortment of pathogenic viruses. Furthermore, our data suggest autophagy-inducing drugs may provide a novel therapeutic strategy for combating lyssavirus infection.


id: cord-273711-bxijla09
author: Zhao, Zhixun
date: 2012-02-17
txt: ./txt/cord-273711-bxijla09.txt
cache: ./cache/cord-273711-bxijla09.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/22340205/
doi: 10.1186/1743-422x-9-48
summary:
  title: RNA interference targeting virion core protein ORF095 inhibits Goatpox virus replication in Vero cells When Vero cells were transfected with shRNA expression vectors (p61/GFP, p70/GFP, p165/GFP and p296/GFP) and then infected with GTPV, GTPV-ORF095-70 was found to be the most effective inhibition site in decreasing cytopathic effect (CPE) induced by GTPV. These results suggest that the siRNA generated by in vitro transcription effectively and specifically inhibit the expression of GTPV ORF095 conserved regions in BHK-21 cells. To investigate whether or not knockout of ORF095 relieves cytopathic effect (CPE) induced by GTPV, Vero cells were transfected by plasmids expressing ORF095 protein-targeted shRNAs (p61/GFP, p70/GFP, p165/GFP and p296/GFP), respectively. Therefore, ORF095 gene is a good target to suppress GTPV replication by RNAi. In conclusion, this study demonstrates that vector-based shRNA methodology can effectively inhibit GTPV replication on Vero cells. RNA interference targeting virion core protein ORF095 inhibits Goatpox virus replication in Vero cells

title:
  RNA interference targeting virion core protein ORF095 inhibits Goatpox virus replication in Vero cells

abstract:
  BACKGROUND: Goatpox is an economically important disease in goat and sheep-producing areas of the world. Many vaccine strategies developed to control the disease are not yet completely successful. Hairpin expression vectors have been used to induce gene silencing in a large number of studies on viruses. However, none of these studies has been attempted to study GTPV. In the interest of exploiting improved methods to control goat pox, it is participated that RNAi may provide effective protection against GTPV. In this study we show the suppression of Goatpox virus (GTPV) replication via knockdown of virion core protein using RNA interference. RESULTS: Four short interfering RNA (siRNA) sequences (siRNA-61, siRNA-70, siRNA-165 and siRNA-296) against a region of GTPV ORF095 were selected. Sense and antisense siRNA-encoding sequences separated by a hairpin loop sequence were designed as short hairpin RNA (shRNA) expression cassettes under the control of a human U6 promoter. ORF095 amplicon was generated using PCR, and then cloned into pEGFP-N1 vector, named as p095/EGFP. p095/EGFP and each of the siRNA expression cassettes (p61, p70, p165 and p296) were co-transfected into BHK-21 cells. Fluorescence detection, flow cytometric analysis, retro transcription PCR (RT-PCR) and real time PCR were used to check the efficiency of RNAi. The results showed that the ORF095-specific siRNA-70 effectively down-regulated the expression of ORF095. When Vero cells were transfected with shRNA expression vectors (p61/GFP, p70/GFP, p165/GFP and p296/GFP) and then infected with GTPV, GTPV-ORF095-70 was found to be the most effective inhibition site in decreasing cytopathic effect (CPE) induced by GTPV. The results presented here indicated that DNA-based siRNA could effectively inhibit the replication of GTPV (approximately 463. 5-fold reduction of viral titers) on Vero cells. CONCLUSIONS: This study demonstrates that vector-based shRNA methodology can effectively inhibit GTPV replication on Vero cells. Simultaneously, this work represents a strategy for controlling goatpox, potentially facilitating new experimental approaches in the analysis of both viral and cellular gene functions during of GTPV infection.


id: cord-274210-uaaj66xq
author: Ding, Ning
date: 2017-02-28
txt: ./txt/cord-274210-uaaj66xq.txt
cache: ./cache/cord-274210-uaaj66xq.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.3389/fcimb.2017.00056
doi: 10.3389/fcimb.2017.00056
summary:
  These results show that PHEV infection induces atypical autophagy and causes the appearance of autophagosomes but blocks the fusion with lysosomes, which is necessary for the replication of PHEV in nerve cells. In this study, we confirmed that PHEV infection induces atypical autophagy and leads to the accumulation of autophagosomes while blocking their fusion with lysosomes, which creates conditions for the virus to replicate within nerve cells. In order to examine whether the autophagy is associated with viral replication in PHEV infection, we performed subcellular localization of viral proteins and LC3 or LAMP on PHEVinfected cells for the first time. In expect to recognize the effect of autophagy induced by PHEV infection on viral replication, we treated the Neuro-2a cells with 3-MA, the autophagic inhibitor targeting a class III phosphatidylinositol-3-kinase (PI3K) (Petiot et al., 2000) .

title:
  Induction of Atypical Autophagy by Porcine Hemagglutinating Encephalomyelitis Virus Contributes to Viral Replication

abstract:
  Autophagy is a basic biological metabolic process involving in intracellular membrane transport pathways that recycle cellular components and eliminate intracellular microorganisms within the lysosome. Autophagy also plays an important part in virus infection and propagation. However, some pathogens, including viruses, have evolved unique trick to escape or exploit autophagy. This study explores the mechanism of autophagy induction by porcine hemagglutinating encephalomyelitis virus (PHEV) in Neuro-2a cells, and examines the role of autophagy in PHEV replication. PHEV triggered autophagy in Neuro-2a cells is dependent on the presence of bulk double- or single-membrane vacuoles, the accumulation of GFP-LC3 fluorescent dots, and the LC3 lipidation. In addition, PHEV induced an incomplete autophagic effect because the degradation level of p62 did not change in PHEV-infected cells. Further validation was captured using LysoTracker and lysosome-associated membrane protein by indirect immunofluorescence labeling in PHEV-infected cells. We also investigated the change in viral replication by pharmacological experiments with the autophagy inducer rapamycin or the autophagy inhibitor 3-MA, and the lysosomal inhibitor chloroquine (CQ). Suppression of autophagy by 3-MA increased viral replication, compared with the mock treatment, while promoting of autophagy by rapamycin reduced PHEV replication. CQ treatment enhanced the LC3 lipidation in PHEV-infected Neuro-2a cells but lowered PHEV replication. These results show that PHEV infection induces atypical autophagy and causes the appearance of autophagosomes but blocks the fusion with lysosomes, which is necessary for the replication of PHEV in nerve cells.


id: cord-275307-d7htyfcl
author: Gaglia, Marta Maria
date: 2015-12-08
txt: ./txt/cord-275307-d7htyfcl.txt
cache: ./cache/cord-275307-d7htyfcl.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/26646420/
doi: 10.1371/journal.ppat.1005305
summary:
  Development of a novel bioinformatics pipeline to detect highconfidence SOX cleavage sites across the transcriptome following PARE Prior analyses of individual mRNAs indicated that the KSHV RNase SOX cuts at specific locations within the RNA, in a manner dependent on the sequence surrounding the cleavage site [5] . This example shows the expected distribution for a cut site followed by exonucleolytic degradation due PARE libraries from two replicates of SOX-expressing or GFP control cells and extracted the 5'' end of each mapped read, which represents the cleavage site (S1 Table) . Indeed, the sequences flanking the set of reproducible SOX cut sites (identified with a confidence level of 99.99%) were a closer match to the motif compared to those surrounding GFP-specific fragment ends, as shown by the distribution of the log likelihood scores (Fig 6A) .

title:
  Transcriptome-Wide Cleavage Site Mapping on Cellular mRNAs Reveals Features Underlying Sequence-Specific Cleavage by the Viral Ribonuclease SOX

abstract:
  Many viruses express factors that reduce host gene expression through widespread degradation of cellular mRNA. An example of this class of proteins is the mRNA-targeting endoribonuclease SOX from the gamma-herpesvirus Kaposi’s sarcoma-associated herpesvirus (KSHV). Previous studies indicated that cleavage of messenger RNAs (mRNA) by SOX occurs at specific locations defined by the sequence of the target RNA, which is at odds with the down-regulation of a large portion of cellular transcripts. In this study, we address this paradox by using high-throughput sequencing of cleavage intermediates combined with a custom bioinformatics-based analysis pipeline to identify SOX cleavage sites across the mRNA transcriptome. These data, coupled with targeted mutagenesis, reveal that while cleavage sites are specific and reproducible, they are defined by a degenerate sequence motif containing a small number of conserved residues rather than a strong consensus sequence. This degenerate element is well represented in both human and KSHV mRNA, and its presence correlates with RNA destabilization by SOX. This represents a new endonuclease targeting strategy, in which use of a degenerate targeting element enables RNA cleavage at specific locations without restricting the range of targets. Furthermore, it shows that strong target selectivity can be achieved without a high degree of sequence specificity.


id: cord-280454-etf32afd
author: Moustaqil, Mehdi
date: 2020-06-05
txt: ./txt/cord-280454-etf32afd.txt
cache: ./cache/cord-280454-etf32afd.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1101/2020.06.05.135699
doi: 10.1101/2020.06.05.135699
summary:
  title: SARS-CoV-2 proteases cleave IRF3 and critical modulators of inflammatory pathways (NLRP12 and TAB1): implications for disease presentation across species and the search for reservoir hosts Direct cleavage of IRF3 by NSP3 could explain the blunted TypeI IFN response seen during SARS-CoV-2 infections while NSP5 mediated cleavage of NLRP12 and TAB1 point to a molecular mechanism for enhanced production of IL-6 and inflammatory response observed in COVID-19 patients. In this report, we show that the viral proteases PLpro and 3CLpro of SARS-CoV2 lead to the in-vitro proteolytic cleavage of three important proteins of the host immune response: IRF3, TAB1 and NLRP12 (Fig. 1B) . The presence of the five human-like cleavage sites for IRF3, TAB1 and NLRP12 in a single species shows that it is possible that the SARS viruses could have gained the new functionality of cleaving these Human Innate Immune Proteins in a single reservoir host, potentially in Myotis Davidii.

title:
  SARS-CoV-2 proteases cleave IRF3 and critical modulators of inflammatory pathways (NLRP12 and TAB1): implications for disease presentation across species and the search for reservoir hosts

abstract:
  The genome of SARS-CoV-2 (SARS2) encodes for two viral proteases (NSP3/ papain-like protease and NSP5/ 3C-like protease or major protease) that are responsible for cleaving viral polyproteins for successful replication. NSP3 and NSP5 of SARS-CoV (SARS1) are known interferon antagonists. Here, we examined whether the protease function of SARS2 NSP3 and NSP5 target proteins involved in the host innate immune response. We designed a fluorescent based cleavage assay to rapidly screen the protease activity of NSP3 and NSP5 on a library of 71 human innate immune proteins (HIIPs), covering most pathways involved in human innate immunity. By expressing each of these HIIPs with a genetically encoded fluorophore in a cell-free system and titrating in the recombinant protease domain of NSP3 or NSP5, we could readily detect cleavage of cognate HIIPs on SDS-page gels. We identified 3 proteins that were specifically and selectively cleaved by NSP3 or NSP5: IRF-3, and NLRP12 and TAB1, respectively. Direct cleavage of IRF3 by NSP3 could explain the blunted Type- I IFN response seen during SARS-CoV-2 infections while NSP5 mediated cleavage of NLRP12 and TAB1 point to a molecular mechanism for enhanced production of IL-6 and inflammatory response observed in COVID-19 patients. Surprisingly, both NLRP12 and TAB1 have each two distinct cleavage sites. We demonstrate that in mice, the second cleavage site of NLRP12 is absent. We pushed this comparative alignment of IRF-3 and NLRP12 homologs and show that the lack or presence of cognate cleavage motifs in IRF-3 and NLRP12 could contribute to the presentation of disease in cats and tigers, for example. Our findings provide an explanatory framework for in-depth studies into the pathophysiology of COVID-19 and should facilitate the search or development of more effective animal models for severe COVID-19. Finally, we discovered that one particular species of bats, David’s Myotis, possesses the five cleavage sites found in humans for NLRP12, TAB1 and IRF3. These bats are endemic from the Hubei province in China and we discuss its potential role as reservoir for the evolution of SARS1 and SASR2.


id: cord-299509-7xjdryoq
author: Scholte, Florine E. M.
date: 2013-08-01
txt: ./txt/cord-299509-7xjdryoq.txt
cache: ./cache/cord-299509-7xjdryoq.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/23936484/
doi: 10.1371/journal.pone.0071047
summary:
  Infectious cDNA clones of viruses have become invaluable tools that allow reverse genetics studies to elucidate the contribution of specific amino acids or RNA structures to viraemia, virulence, antigenicity, replication kinetics, interactions with host factors, adaptation to new vectors, and many other aspects of the viral life cycle. To obtain a virus that -in terms of virulence, sensitivity to antiviral compounds, and CHIKV-host interactions -is expected to have the general characteristics of the E1-226V CHIKV strains that were circulating during the 2005-2009 outbreaks, we have constructed a completely synthetic CHIKV cDNA clone based on the consensus sequence of the aligned genomes of these recent isolates. Indirect immunofluorescence analysis of Vero E6 cells infected with CHIKV LS3, LS3-GFP, or ITA07-RA1 at various time points showed that the localization and expression kinetics of E2 and dsRNA were similar for the natural isolate and the synthetic viruses (Fig. 6) .

title:
  Characterization of Synthetic Chikungunya Viruses Based on the Consensus Sequence of Recent E1-226V Isolates

abstract:
  Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that re-emerged in 2004 and has caused massive outbreaks in recent years. The lack of a licensed vaccine or treatment options emphasize the need to obtain more insight into the viral life cycle and CHIKV-host interactions. Infectious cDNA clones are important tools for such studies, and for mechanism of action studies on antiviral compounds. Existing CHIKV cDNA clones are based on a single genome from an individual clinical isolate, which is expected to have evolved specific characteristics in response to the host environment, and possibly also during subsequent cell culture passaging. To obtain a virus expected to have the general characteristics of the recent E1-226V CHIKV isolates, we have constructed a new CHIKV full-length cDNA clone, CHIKV LS3, based on the consensus sequence of their aligned genomes. Here we report the characterization of this synthetic virus and a green fluorescent protein-expressing variant (CHIKV LS3-GFP). Their characteristics were compared to those of natural strain ITA07-RA1, which was isolated during the 2007 outbreak in Italy. In cell culture the synthetic viruses displayed phenotypes comparable to the natural isolate, and in a mouse model they caused lethal infections that were indistinguishable from infections with a natural strain. Compared to ITA07-RA1 and clinical isolate NL10/152, the synthetic viruses displayed similar sensitivities to several antiviral compounds. 3-deaza-adenosine was identified as a new inhibitor of CHIKV replication. Cyclosporin A had no effect on CHIKV replication, suggesting that cyclophilins -opposite to what was found for other +RNA viruses- do not play an essential role in CHIKV replication. The characterization of the consensus sequence-based synthetic viruses and their comparison to natural isolates demonstrated that CHIKV LS3 and LS3-GFP are suitable and representative tools to study CHIKV-host interactions, screen for antiviral compounds and unravel their mode of action.


id: cord-307067-cpc1yefj
title: A single dose of ChAdOx1 MERS provides protective immunity in rhesus macaques
author: van Doremalen, Neeltje
date: 2020-06-10
txt: ./txt/cord-307067-cpc1yefj.txt
cache: ./cache/cord-307067-cpc1yefj.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1126/sciadv.aba8399
doi: 10.1126/sciadv.aba8399
summary:
  For Middle East respiratory syndrome coronavirus (MERS-CoV), we show that rhesus macaques seroconverted rapidly after a single intramuscular vaccination with ChAdOx1 MERS. A prime-boost regimen of ChAdOx1 MERS boosted antibody titers, and viral replication was completely absent from the respiratory tract tissue of these rhesus macaques. Viral load was higher for lower respiratory tract tissue obtained from animals vaccinated with ChAdOx1 GFP (n = 6) than from animals receiving a prime-only (n = 6) or a prime-boost regimen of ChAdOx1 MERS (n = 2) (Fig. 4B ). Notably, antigenic differences have been reported between S proteins from the Middle East and Africa (8), potentially affecting the efficacy of a vaccine based In conclusion, we show that a single vaccination with ChAdOx1 MERS results in protection against disease progression and virus replication associated with MERS-CoV challenge in the rhesus macaque, and a prime-boost regimen reduced viral replication further.

abstract:
  Developing a vaccine to protect against the lethal effects of the many strains of coronavirus is critical given the current global pandemic. For Middle East respiratory syndrome coronavirus (MERS-CoV), we show that rhesus macaques seroconverted rapidly after a single intramuscular vaccination with ChAdOx1 MERS. The vaccine protected against respiratory injury and pneumonia and reduced viral load in lung tissue by several orders of magnitude. MERS-CoV replication in type I and II pneumocytes of ChAdOx1 MERS–vaccinated animals was absent. A prime-boost regimen of ChAdOx1 MERS boosted antibody titers, and viral replication was completely absent from the respiratory tract tissue of these rhesus macaques. We also found that antibodies elicited by ChAdOx1 MERS in rhesus macaques neutralized six different MERS-CoV strains. Transgenic human dipeptidyl peptidase 4 mice vaccinated with ChAdOx1 MERS were completely protected against disease and lethality for all different MERS-CoV strains. The data support further clinical development of ChAdOx1 MERS.


id: cord-315483-l6dm82pp
author: Santhakumar, Diwakar
date: 2018-05-01
txt: ./txt/cord-315483-l6dm82pp.txt
cache: ./cache/cord-315483-l6dm82pp.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/29717152/
doi: 10.1038/s41598-018-24905-y
summary:
  To confirm the sequence of the cDNA and to identify the genomic structure of the identified IFIT gene, the transcript was amplified from RNA extracted from NDV-infected CEFs. Complete sequence analysis of the gene revealed an open reading frame (ORF) of 1440 bps (479 amino acids excluding stop codon) with high sequence identity (48% and 67%) with the human and duck IFIT5 proteins, respectively ( Supplementary Fig. 1B) . The levels of chIFN-β gene induction was proportional to the NDV replication (Fig. 2D) , therefore, it can be inferred that the virus-induced expression of chicken IFIT5 is IFN-dependent and that chIFIT5 is an early-ISG with capacity to modulate initial steps of virus life cycle. To compare anti-viral effects of chIFIT5 (only IFIT gene identified in chickens so far) with its orthologous and homologous human proteins, huIFIT1, huIFIT2, huIFIT3 and huIFIT5 were expressed in chicken cells and antiviral affect was monitored (Fig. 5A ).

title:
  Chicken Interferon-induced Protein with Tetratricopeptide Repeats 5 Antagonizes Replication of RNA Viruses

abstract:
  The intracellular actions of interferon (IFN)-regulated proteins, including IFN-induced proteins with tetratricopeptide repeats (IFITs), attribute a major component of the protective antiviral host defense. Here we applied genomics approaches to annotate the chicken IFIT locus and currently identified a single IFIT (chIFIT5) gene. The profound transcriptional level of this effector of innate immunity was mapped within its unique cis-acting elements. This highly virus- and IFN-responsive chIFIT5 protein interacted with negative sense viral RNA structures that carried a triphosphate group on its 5′ terminus (ppp-RNA). This interaction reduced the replication of RNA viruses in lentivirus-mediated IFIT5-stable chicken fibroblasts whereas CRISPR/Cas9-edited chIFIT5 gene knockout fibroblasts supported the replication of RNA viruses. Finally, we generated mosaic transgenic chicken embryos stably expressing chIFIT5 protein or knocked-down for endogenous chIFIT5 gene. Replication kinetics of RNA viruses in these transgenic chicken embryos demonstrated the antiviral potential of chIFIT5 in ovo. Taken together, these findings propose that IFIT5 specifically antagonize RNA viruses by sequestering viral nucleic acids in chickens, which are unique in innate immune sensing and responses to viruses of both poultry and human health significance.


id: cord-316814-9fv9xrln
title: Use of GFP to Investigate Expression of Plant-Derived Vaccines
author: Li, Hong-Ye
date: 2009
txt: ./txt/cord-316814-9fv9xrln.txt
cache: ./cache/cord-316814-9fv9xrln.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1007/978-1-59745-559-6_19
doi: 10.1007/978-1-59745-559-6_19
summary:
  Agroinfiltration is a more recent technique that can be applied to investigate transient expression in plant cells by which an Agrobacterium liquid culture is infiltrated into intact plant leaves (7) . By simple infiltration of Agrobacterium cells carrying appropriate gene constructs into tobacco plants leaves, transient expression assays can be performed within 3 days without using expensive instruments or complicated procedures. Two days after agroinfiltration, expression and subcellular localization of the GFP fusion proteins in tobacco leaves can be determined by simple observation under fluorescence or confocal laser scanning microscopy. 7. Generate plasmid pCV12 (Fig. 1 ) or similar nuclear transformation vector for expression of a fusion protein consisting of the SARS-CoV S1 and GFP. Production of tobacco leaves transiently expressing a protein fusion consisting of the SARS-CoV S1 protein fused with the GFP was carried out using Agrobacterium-mediated transformation with plasmid pCV12.

abstract:
  Plants are low-cost bioreactors for the production of various biopharmaceuticals including oral vaccines. Plant-derived oral vaccines are potentially useful in combating viral infections involving mucosal immunity. Transgenic plants have been generated to successfully produce mucosal vaccines against cholera, hepatitis B, foot-and-mouth disease, and Norwalk virus. As a first step toward the generation of oral vaccines against the severe acute respiratory syndrome coronavirus (SARS-CoV), we have expressed a recombinant S1 protein of the SARS-CoV in transformed tobacco. Since plant transformation and regeneration of stable transformants require considerable time, we initially used a green fluorescent protein (GFP) to tag the antigen in transient expression. GFP was fused to the carboxy-terminus of S1 for expression of S1-GFP to show expression of recombinant S1 by agroinfiltration of tobacco leaves. The GFP tag enables a relatively quick confirmation of antigen expression in plant cells by fluorescent microscopy. Such analysis using GFP that precedes stable plant transformation will enable the rapid screening of multiple constructs to attain optimal recombinant protein expression. Furthermore, this approach determines the subcellular localization of the recombinant protein in plant cells, providing information on optimal subcellular targeting for production in plant bioreactors.


id: cord-317142-qd61qvch
author: Muramatsu, Tomonari
date: 2013-03-27
txt: ./txt/cord-317142-qd61qvch.txt
cache: ./cache/cord-317142-qd61qvch.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1111/febs.12222
doi: 10.1111/febs.12222
summary:
  In vitro 3CL pro autoprocessing system First we developed a SARS 3CL protease autoprocessing system by use of the Escherichia coli cell-free protein synthesis system, with the N-and C-terminal 10 amino acid pro-sequences accompanied by an S tag and a His tag, respectively (Fig. 1A) . Over the course of the cell-free protein synthesis reaction (30°C, 4 h), the N-and C-terminal processing sites of the catalytically active construct (wild-type, WT in Fig. 1A) were completely cleaved by the enzyme''s endogenous proteolytic activity, as shown in Fig. 1B ,C (WT). To investigate the activities of the pro-forms of SARS-3CL pro in detail, we developed a ''trans-cleaving assay'', using a substrate in which the core region of the 3CL protease was exchanged with GFP (green fluorescent protein) ( Fig. 2A) . We estimated the activity of the enzyme (or pro-enzyme) toward the N-and C-terminal processing sites in the GFP substrate on the basis of the dilution ratio at which 50% cleavage was achieved (Fig. 2C) .

title:
  Autoprocessing mechanism of severe acute respiratory syndrome coronavirus 3C‐like protease (SARS‐CoV 3CL (pro)) from its polyproteins

abstract:
  Like many other RNA viruses, severe acute respiratory syndrome coronavirus (SARS‐CoV) produces polyproteins containing several non‐structural proteins, which are then processed by the viral proteases. These proteases often exist within the polyproteins, and are excised by their own proteolytic activity (‘autoprocessing’). It is important to investigate the autoprocessing mechanism of these proteases from the point of view of anti‐SARS‐CoV drug design. In this paper, we describe a new method for investigating the autoprocessing mechanism of the main protease (M (pro)), which is also called the 3C‐like protease (3CL (pro)). Using our method, we measured the activities, under the same conditions, of the mature form and pro‐forms with the N‐terminal pro‐sequence, the C‐terminal pro‐sequence or both pro‐sequences, toward the pro‐form with both N‐ and C‐terminal pro‐sequences. The data indicate that the pro‐forms of the enzyme have proteolytic activity, and are stimulated by the same proteolytic activity. The stimulation occurs in two steps, with approximately eightfold stimulation by N‐terminal cleavage, approximately fourfold stimulation by C‐terminal cleavage, and 23‐fold stimulation by the cleavage of both termini, compared to the pro‐form with both the N‐ and C‐terminal pro‐sequences. Such cleavage mainly occurs in a trans manner; i.e. the pro‐form dimer cleaves the monomeric form. The stimulation by N‐terminal pro‐sequence removal is due to the cis (intra‐dimer and inter‐protomer) effect of formation of the new N‐terminus, whereas that by C‐terminal cleavage is due to removal of its trans (inter‐dimer) inhibitory effect. A numerical simulation of the maturation pathway is presented.


id: cord-319517-denczc6t
title: Recent advances in live cell imaging of hepatoma cells
author: Salipalli, Sandeep
date: 2014-07-08
txt: ./txt/cord-319517-denczc6t.txt
cache: ./cache/cord-319517-denczc6t.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1186/1471-2121-15-26
doi: 10.1186/1471-2121-15-26
summary:
  This review aims to provide an overview of the recent developments in the field of marker proteins (bioluminescence and fluorescent) and methodologies (fluorescent resonance energy transfer, fluorescent recovery after photobleaching and proximity ligation assay) employed as to achieve an improved imaging of biological processes in hepatoma cells. The success of live cell imaging relies on various factors including the specific imaging system, climate controlling devices for cultured cells under investigation, construction of recombinant plasmid DNA, transfer and expression of candidate genes and/or fluorescent proteins in mammalian cells. T4SS (Bartonella sp.) 60-70% 50% >70% >60% 50-60% [76] [77] [78] lipofection (1 μg) methods were used for transfection of plasmid encoding fluorescent protein (pEGFP and pEYFP) tagged to human and mouse ADRP genes in Huh-7 cells as to study the pathways of lipid droplet metabolism.

abstract:
  Live cell imaging enables the study of dynamic processes of living cells in real time by use of suitable reporter proteins and the staining of specific cellular structures and/or organelles. With the availability of advanced optical devices and improved cell culture protocols it has become a rapidly growing research methodology. The success of this technique relies mainly on the selection of suitable reporter proteins, construction of recombinant plasmids possessing cell type specific promoters as well as reliable methods of gene transfer. This review aims to provide an overview of the recent developments in the field of marker proteins (bioluminescence and fluorescent) and methodologies (fluorescent resonance energy transfer, fluorescent recovery after photobleaching and proximity ligation assay) employed as to achieve an improved imaging of biological processes in hepatoma cells. Moreover, different expression systems of marker proteins and the modes of gene transfer are discussed with emphasis on the study of lipid droplet formation in hepatocytes as an example.


id: cord-319842-4mnaicki
title: Subversion of Cellular Autophagosomal Machinery by RNA Viruses
author: Jackson, William T
date: 2005-04-26
txt: ./txt/cord-319842-4mnaicki.txt
cache: ./cache/cord-319842-4mnaicki.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/15884975/
doi: 10.1371/journal.pbio.0030156
summary:
  Infection of human cells with poliovirus induces the proliferation of double-membraned cytoplasmic vesicles whose surfaces are used as the sites of viral RNA replication and whose origin is unknown. Here, we explore the role of several constituents of autophagosome machinery in poliovirus-and rhinovirusinfected cells by monitoring: the presence of autophagosomal protein LC3 in virally induced vesicles, the acquisition of colocalization of LC3 and LAMP1 in virally infected cells, the viral induction of punctate structures that stain with monodansylcadaverine (MDC), and the effects of perturbing the autophagosomal pathway pharmacologically and via RNA interference on intracellular and extracellular virus yield. To determine whether the autophagosome-like membranes induced during poliovirus infection perform an antiviral function or facilitate viral replication, we tested the effect on poliovirus yield of pretreating H1-HeLa cells with either tamoxifen or rapamycin, known inducers of autophagy.

abstract:
  Infection of human cells with poliovirus induces the proliferation of double-membraned cytoplasmic vesicles whose surfaces are used as the sites of viral RNA replication and whose origin is unknown. Here, we show that several hallmarks of cellular autophagosomes can be identified in poliovirus-induced vesicles, including colocalization of LAMP1 and LC3, the human homolog of Saccharomyces cerevisiae Atg8p, and staining with the fluorophore monodansylcadaverine followed by fixation. Colocalization of LC3 and LAMP1 was observed early in the poliovirus replicative cycle, in cells infected with rhinoviruses 2 and 14, and in cells that express poliovirus proteins 2BC and 3A, known to be sufficient to induce double-membraned vesicles. Stimulation of autophagy increased poliovirus yield, and inhibition of the autophagosomal pathway by 3-methyladenine or by RNA interference against mRNAs that encode two different proteins known to be required for autophagy decreased poliovirus yield. We propose that, for poliovirus and rhinovirus, components of the cellular machinery of autophagosome formation are subverted to promote viral replication. Although autophagy can serve in the innate immune response to microorganisms, our findings are inconsistent with a role for the induced autophagosome-like structures in clearance of poliovirus. Instead, we argue that these double-membraned structures provide membranous supports for viral RNA replication complexes, possibly enabling the nonlytic release of cytoplasmic contents, including progeny virions, from infected cells.


id: cord-324720-acei0pn8
title: Inflammatory monocytes damage the hippocampus during acute picornavirus infection of the brain
author: Howe, Charles L
date: 2012-03-09
txt: ./txt/cord-324720-acei0pn8.txt
cache: ./cache/cord-324720-acei0pn8.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/22405261/
doi: 10.1186/1742-2094-9-50
summary:
  METHODS: The identity of brain-infiltrating leukocytes was determined using microscopy and flow cytometry at several acute time points following intracranial infection of mice with the Theiler''s murine encephalomyelitis virus. Likewise, the population of CD45 hi cells could be distinguished by surface F4/80 expression, with some F4/80 + Figure 1 Histological evidence of rapid immune cell infiltration into the hippocampus of mice acutely infected with Theiler''s murine encephalomyelitis virus (TMEV). We were unable to identify a population of inflammatory monocytes in the blood in the infected mice, suggesting either that these cells are not circulating at this time or that so many have been recruited to the brain that the Figure 2 Flow cytometric assessment of the infiltrate present in the brain of mice acutely infected with Theiler''s murine encephalomyelitis virus (TMEV).

abstract:
  BACKGROUND: Neuropathology caused by acute viral infection of the brain is associated with the development of persistent neurological deficits. Identification of the immune effectors responsible for injuring the brain during acute infection is necessary for the development of therapeutic strategies that reduce neuropathology but maintain immune control of the virus. METHODS: The identity of brain-infiltrating leukocytes was determined using microscopy and flow cytometry at several acute time points following intracranial infection of mice with the Theiler's murine encephalomyelitis virus. Behavioral consequences of immune cell depletion were assessed by Morris water maze. RESULTS: Inflammatory monocytes, defined as CD45(hi)CD11b(++)F4/80(+)Gr1(+)1A8(-), and neutrophils, defined as CD45(hi)CD11b(+++)F4/80(-)Gr1(+)1A8(+), were found in the brain at 12 h after infection. Flow cytometry of brain-infiltrating leukocytes collected from LysM: GFP reporter mice confirmed the identification of neutrophils and inflammatory monocytes in the brain. Microscopy of sections from infected LysM:GFP mice showed that infiltrating cells were concentrated in the hippocampal formation. Immunostaining confirmed that neutrophils and inflammatory monocytes were localized to the hippocampal formation at 12 h after infection. Immunodepletion of inflammatory monocytes and neutrophils but not of neutrophils only resulted in preservation of hippocampal neurons. Immunodepletion of inflammatory monocytes also preserved cognitive function as assessed by the Morris water maze. CONCLUSIONS: Neutrophils and inflammatory monocytes rapidly and robustly responded to Theiler's virus infection by infiltrating the brain. Inflammatory monocytes preceded neutrophils, but both cell types were present in the hippocampal formation at a timepoint that is consistent with a role in triggering hippocampal pathology. Depletion of inflammatory monocytes and neutrophils with the Gr1 antibody resulted in hippocampal neuroprotection and preservation of cognitive function. Specific depletion of neutrophils with the 1A8 antibody failed to preserve neurons, suggesting that inflammatory monocytes are the key effectors of brain injury during acute picornavirus infection of the brain. These effector cells may be important therapeutic targets for immunomodulatory or immunosuppressive therapies aimed at reducing or preventing central nervous system pathology associated with acute viral infection.


id: cord-330176-1ugzkf22
author: Weeratunga, Prasanna
date: 2016-01-05
txt: ./txt/cord-330176-1ugzkf22.txt
cache: ./cache/cord-330176-1ugzkf22.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/26727903/
doi: 10.1007/s12275-016-5555-4
summary:
  In vitro, an effective dose of Angelica tenuissima Nakai markedly inhibited the replication of Influenza A virus (PR8), Vesicular stomatitis virus (VSV), Herpes simplex virus (HSV), Coxsackie virus, and Enterovirus (EV-71) on epithelial (HEK293T/HeLa) and immune (RAW264.7) cells. To investigate the antiviral effects in immune cells, we first assessed the replication of divergent GFP-expressing viruses that were treated or untreated with cytotoxic-free (data not shown) Angelica tenuissima Nakai in RAW264.7 cells. These results suggest that the aqueous extract of Angelica tenuissima Nakai can induce the secretion of IFNs and pro-inflammatory cytokines, which can stimulate the cellular antiviral state for inhibition of viral replication. The aqueous extract of Angelica tenuissima Nakai inhibit the diverse viral infection through the induction of Type I IFN signaling and pro-inflammatory cytokines, leading to an antiviral state in epithelial and immune cells.

title:
  RETRACTED ARTICLE: Interferon-mediated antiviral activities of Angelica tenuissima Nakai and its active components

abstract:
  Angelica tenuissima Nakai is a widely used commodity in traditional medicine. Nevertheless, no study has been conducted on the antiviral and immune-modulatory properties of an aqueous extract of Angelica tenuissima Nakai. In the present study, we evaluated the antiviral activities and the mechanism of action of an aqueous extract of Angelica tenuissima Nakai both in vitro and in vivo. In vitro, an effective dose of Angelica tenuissima Nakai markedly inhibited the replication of Influenza A virus (PR8), Vesicular stomatitis virus (VSV), Herpes simplex virus (HSV), Coxsackie virus, and Enterovirus (EV-71) on epithelial (HEK293T/HeLa) and immune (RAW264.7) cells. Such inhibition can be described by the induction of the antiviral state in cells by antiviral, IFNrelated gene induction and secretion of IFNs and pro-inflammatory cytokines. In vivo, Angelica tenuissima Nakai treated BALB/c mice displayed higher survivability and lower lung viral titers when challenged with lethal doses of highly pathogenic influenza A subtypes (H1N1, H5N2, H7N3, and H9N2). We also found that Angelica tenuissima Nakai can induce the secretion of IL-6, IFN-λ, and local IgA in bronchoalveolar lavage fluid (BALF) of Angelica tenuissima Nakai treated mice, which correlating with the observed prophylactic effects. In HPLC analysis, we found the presence of several compounds in the aqueous fraction and among them; we evaluated antiviral properties of ferulic acid. Therefore, an extract of Angelica tenuissima Nakai and its components, including ferulic acid, play roles as immunomodulators and may be potential candidates for novel anti-viral/anti-influenza agents.


id: cord-331414-i0oxm5mr
author: Kautz, Tiffany F.
date: 2020-06-19
url: https://www.ncbi.nlm.nih.gov/pubmed/32575413/
doi: 10.3390/v12060660
title:
  A Low Fidelity Virus Shows Increased Recombination during the Removal of an Alphavirus Reporter Gene

abstract:
  Reporter genes for RNA viruses are well-known to be unstable due to putative RNA recombination events that excise inserted nucleic acids. RNA recombination has been demonstrated to be co-regulated with replication fidelity in alphaviruses, but it is unknown how recombination events at the minority variant level act, which is important for vaccine and trans-gene delivery design. Therefore, we sought to characterize the removal of a reporter gene by a low-fidelity alphavirus mutant over multiple replication cycles. To examine this, GFP was inserted into TC-83, a live-attenuated vaccine for the alphavirus Venezuelan equine encephalitis virus, as well as a low-fidelity variant of TC-83, and passaged until fluorescence was no longer observed. Short-read RNA sequencing using ClickSeq was performed to determine which regions of the viral genome underwent recombination and how this changed over multiple replication cycles. A rapid removal of the GFP gene was observed, where minority variants in the virus population accumulated small deletions that increased in size over the course of passaging. Eventually, these small deletions merged to fully remove the GFP gene. The removal was significantly enhanced during the passaging of low-fidelity TC-83, suggesting that increased levels of recombination are a defining characteristic of this mutant.


id: cord-332881-mkm4ygh6
author: Kirchhoff, Jana
date: 2014-02-18
txt: ./txt/cord-332881-mkm4ygh6.txt
cache: ./cache/cord-332881-mkm4ygh6.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/24548739/
doi: 10.1186/1297-9716-45-20
summary:
  We investigated the susceptibility of bovine airway epithelial cells (BAEC) to infection by the three major viruses associated with the BRDC: bovine respiratory syncytial virus (BRSV), bovine herpesvirus type 1 (BHV-1) and bovine parainfluenza virus type 3 (BPIV3). For this purpose, two culture systems for well-differentiated BAEC were used: the air-liquid interface (ALI) system, where filter-grown BAEC differentiate into a pseudostratified respiratory epithelium and precision-cut lung slices (PCLS) where BAEC are maintained in the original tissue organisation. We have reported recently that well-differentiated respiratory epithelial cells are susceptible to infection by BPIV3 whereas they are rather resistant to infection by BRSV [7] . BPIV3 efficiently infected the airway epithelial cells using sialic acids on the apical surface as a receptor determinant for virus entry from the apical side. When BHV-1-GFP was applied to the apical side of the well-differentiated airway epithelial cells, infected cells were detected only occasionally.

title:
  Three viruses of the bovine respiratory disease complex apply different strategies to initiate infection

abstract:
  Bovine respiratory disease complex (BRDC) is the major cause of serious respiratory tract infections in calves. The disease is multifactorial, with either stress or reduced immunity allowing several pathogens to emerge. We investigated the susceptibility of bovine airway epithelial cells (BAEC) to infection by the three major viruses associated with the BRDC: bovine respiratory syncytial virus (BRSV), bovine herpesvirus type 1 (BHV-1) and bovine parainfluenza virus type 3 (BPIV3). For this purpose, two culture systems for well-differentiated BAEC were used: the air-liquid interface (ALI) system, where filter-grown BAEC differentiate into a pseudostratified respiratory epithelium and precision-cut lung slices (PCLS) where BAEC are maintained in the original tissue organisation. Comparative infection studies demonstrated that entry and release of BPIV3 occurred specifically via the apical membrane with ciliated cells being the major target cells. By contrast, airway epithelial cells were largely resistant to infection by BHV-1. When the epithelial barrier was abolished by opening tight junctions or by injuring the cell monolayer, BHV-1 infected mainly basal cells. Respiratory epithelial cells were also refractory to infection by BRSV. However, this virus infected neither differentiated epithelial cells nor basal cells when the integrity of the epithelial barrier was destroyed. In contrast to cells of the airway epithelium, subepithelial cells were susceptible to infection by BRSV. Altogether, these results indicate that the three viruses of the same disease complex follow different strategies to interact with the airway epithelium. Possible entry mechanisms are discussed.


id: cord-334855-s0ci3r8w
title: Novel Antiviral Activities of Obatoclax, Emetine, Niclosamide, Brequinar, and Homoharringtonine
author: Andersen, Petter I.
date: 2019-10-18
txt: ./txt/cord-334855-s0ci3r8w.txt
cache: ./cache/cord-334855-s0ci3r8w.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.3390/v11100964
doi: 10.3390/v11100964
summary:
  Here, we identified novel activities of obatoclax and emetine against herpes simplex virus type 2 (HSV-2), echovirus 1 (EV1), human metapneumovirus (HMPV) and Rift Valley fever virus (RVFV) in cell cultures. The expected response of the emetine-obatoclax drug combination on the viability of FLUAV-and mock-infected RPE cells was calculated using Bliss reference model [29] . After the initial screening, we identified four compounds (obatoclax, emetine, niclosamide and ganciclovir) that at none-cytotoxic concentrations rescued cells from virus-mediated death (Figure 2A) . Table S1 : Compounds, their suppliers and catalogue numbers; Table S2 : Developmental status of broad-spectrum antivirals used in the study; Table S3 : Human viruses and associated diseases; Figure S1 : The expected response of the emetine-obatoclax drug combination on the viability of FLUAV-and mock-infected RPE cells, as measured with the CTG assay using the Bliss reference model; Figure S2 

abstract:
  Viruses are the major causes of acute and chronic infectious diseases in the world. According to the World Health Organization, there is an urgent need for better control of viral diseases. Repurposing existing antiviral agents from one viral disease to another could play a pivotal role in this process. Here, we identified novel activities of obatoclax and emetine against herpes simplex virus type 2 (HSV-2), echovirus 1 (EV1), human metapneumovirus (HMPV) and Rift Valley fever virus (RVFV) in cell cultures. Moreover, we demonstrated novel activities of emetine against influenza A virus (FLUAV), niclosamide against HSV-2, brequinar against human immunodeficiency virus 1 (HIV-1), and homoharringtonine against EV1. Our findings may expand the spectrum of indications of these safe-in-man agents and reinforce the arsenal of available antiviral therapeutics pending the results of further in vitro and in vivo tests.


id: cord-338811-2bi2edcw
title: Imaging-Based Reporter Systems to Define CVB-Induced Membrane Remodeling in Living Cells
author: Lennemann, Nicholas J.
date: 2020-09-25
txt: ./txt/cord-338811-2bi2edcw.txt
cache: ./cache/cord-338811-2bi2edcw.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.3390/v12101074
doi: 10.3390/v12101074
summary:
  To define the dynamic process of enterovirus membrane remodeling of major secretory pathway organelles, we have developed plasmid-based reporter systems that utilize viral protease-dependent release of a nuclear-localized fluorescent protein from the endoplasmic reticulum (ER) membrane during infection, while retaining organelle-specific fluorescent protein markers such as the ER and Golgi. To monitor enterovirus infection in real-time, we adapted cell-based reporter methodologies previously used for flaviviruses and hepatitis C virus that rely on viral protease cleavage-dependent translocation of a membrane-anchored cytoplasmic fluorescent proteins to the nucleus [27] [28] [29] . CVB infection of RepER expressing U2OS cells resulted in a clear translocation of GFP-NLS reporter to the nucleus and eventual dispersal due to cell lysis, while the mCherry-KDEL was maintained in the membranous structure of the ER (Video S1 and Figure 2a) . Live-cell imaging of CVB infected cells expressing these reporters allowed for the real-time visualization of virus-induced changes to the host cell, including the collapse of the peripheral ER network and loss of Golgi integrity.

abstract:
  Enteroviruses manipulate host membranes to form replication organelles, which concentrate viral and host factors to allow for efficient replication. However, this process has not been well-studied in living cells throughout the course of infection. To define the dynamic process of enterovirus membrane remodeling of major secretory pathway organelles, we have developed plasmid-based reporter systems that utilize viral protease-dependent release of a nuclear-localized fluorescent protein from the endoplasmic reticulum (ER) membrane during infection, while retaining organelle-specific fluorescent protein markers such as the ER and Golgi. This system thus allows for the monitoring of organelle-specific changes induced by infection in real-time. Using long-term time-lapse imaging of living cells infected with coxsackievirus B3 (CVB), we detected reporter translocation to the nucleus beginning ~4 h post-infection, which correlated with a loss of Golgi integrity and a collapse of the peripheral ER. Lastly, we applied our system to study the effects of a calcium channel inhibitor, 2APB, on virus-induced manipulation of host membranes. We found that 2APB treatment had no effect on the kinetics of infection or the percentage of infected cells. However, we observed aberrant ER structures in CVB-infected cells treated with 2APB and a significant decrease in viral-dependent cell lysis, which corresponded with a decrease in extracellular virus titers. Thus, our system provides a tractable platform to monitor the effects of inhibitors, gene silencing, and/or gene editing on viral manipulation of host membranes, which can help determine the mechanism of action for antivirals.


id: cord-342189-ya05m58o
title: SARS-CoV-2 disrupts splicing, translation, and protein trafficking to suppress host defenses
author: Banerjee, Abhik K.
date: 2020-10-08
txt: ./txt/cord-342189-ya05m58o.txt
cache: ./cache/cord-342189-ya05m58o.txt
extension: .txt
mime: text/plain
url: https://api.elsevier.com/content/article/pii/S0092867420313106
doi: 10.1016/j.cell.2020.10.004
summary:
  Here, we comprehensively define the interactions between each SARS-CoV-2 protein and human RNAs. We show that 10 viral proteins form highly specific interactions with mRNAs or ncRNAs, including those involved in progressive steps of host cell protein production. We show J o u r n a l P r e -p r o o f 5 that NSP16 binds to the mRNA recognition domains of the U1 and U2 RNA components of the spliceosome and acts to suppress global mRNA splicing in SARS-CoV-2-infected human cells. We identified several pathogenic functions of SARS-CoV-2 in human cells -including global inhibition of host mRNA splicing, protein translation, and membrane protein trafficking -and described the molecular mechanisms by which the virus acts to disrupt these essential cell processes.

abstract:
  SARS-CoV-2 is a recently identified coronavirus that causes the respiratory disease known as COVID-19. Despite the urgent need, we still do not fully understand the molecular basis of SARS-CoV-2 pathogenesis. Here, we comprehensively define the interactions between SARS-CoV-2 proteins and human RNAs. NSP16 binds to the mRNA recognition domains of the U1 and U2 splicing RNAs and acts to suppress global mRNA splicing upon SARS-CoV-2 infection. NSP1 binds to 18S ribosomal RNA in the mRNA entry channel of the ribosome and leads to global inhibition of mRNA translation upon infection. Finally, NSP8 and NSP9 bind to the 7SL RNA in the Signal Recognition Particle and interfere with protein trafficking to the cell membrane upon infection. Disruption of each of these essential cellular functions acts to suppress the interferon response to viral infection. Our results uncover a multipronged strategy utilized by SARS-CoV-2 to antagonize essential cellular processes to suppress host defenses.


id: cord-345651-admlzeu4
author: Wang, Gang
date: 2019-03-30
txt: ./txt/cord-345651-admlzeu4.txt
cache: ./cache/cord-345651-admlzeu4.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/30935078/
doi: 10.3390/v11040313
summary:
  Using a novel approach, the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) systems efficiently and rapidly rescued another recombinant virus with a 224-amino-acid deletion in the N-terminal domain of the TGEV Spike gene (S_NTD224), which is analogous to the N-terminal domain of porcine respiratory coronavirus. Homologous recombination was then performed using the ClonExpress II One Step Cloning Kit (Vazyme, Nanjing, Jiangsu, China) according to the manufacturer''s instructions using 200 ng of recycled linearized pTGEV-GFP BAC, 45 ng of PCR products of S_NTD and two pairs of primers (rec-672SF/δS-NTDR and rec-672SR/δS-NTDF) ( Table 2) . To construct an infectious clone of TGEV, six overlapping cDNA fragments designated A to F were generated by reverse transcriptase PCR (RT-PCR) using total RNA extracted from PK-15 cells infected with TGEV WH-1 ( Figure 1A ,B). Complete genomic sequences, a key residue in the spike protein and deletions in nonstructural protein 3b of US strains of the virulent and attenuated coronaviruses, transmissible gastroenteritis virus and porcine respiratory coronavirus

title:
  The N-Terminal Domain of Spike Protein Is Not the Enteric Tropism Determinant for Transmissible Gastroenteritis Virus in Piglets

abstract:
  Transmissible gastroenteritis virus (TGEV) is the etiologic agent of transmissible gastroenteritis in pigs, and the N-terminal domain of TGEV spike protein is generally recognized as both the virulence determinant and enteric tropism determinant. Here, we assembled a full-length infectious cDNA clone of TGEV in a bacterial artificial chromosome. Using a novel approach, the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) systems efficiently and rapidly rescued another recombinant virus with a 224-amino-acid deletion in the N-terminal domain of the TGEV Spike gene (S_NTD224), which is analogous to the N-terminal domain of porcine respiratory coronavirus. S_NTD224 notably affected the TGEV growth kinetics in PK-15 cells but was not essential for recombinant virus survival. In animal experiments with 13 two-day-old piglets, the TGEV recombinant viruses with/without S_NTD224 deletion induced obvious clinical signs and mortality. Together, our results directly demonstrated that S_NTD224 of TGEV mildly influenced TGEV virulence but was not the enteric tropism determinant and provide new insights for the development of a new attenuated vaccine against TGEV. Importantly, the optimized reverse genetics platform used in this study will simplify the construction of mutant infectious clones and help accelerate progress in coronavirus research.


id: cord-346554-a98pjtxs
title: Inhibitory effects of bee venom and its components against viruses in vitro and in vivo
author: Uddin, Md Bashir
date: 2016-11-26
txt: ./txt/cord-346554-a98pjtxs.txt
cache: ./cache/cord-346554-a98pjtxs.txt
extension: .txt
mime: text/plain
url: https://www.ncbi.nlm.nih.gov/pubmed/27888461/
doi: 10.1007/s12275-016-6376-1
summary:
  Co-incubation of non-cytotoxic amounts of BV and MLT, the main component of BV, significantly inhibited the replication of enveloped viruses such as Influenza A virus (PR8), Vesicular Stomatitis Virus (VSV), Respiratory Syncytial Virus (RSV), and Herpes Simplex Virus (HSV). To compare the antiviral effects of BV to VSV-GFP infection, the levels of viral replication (which reflected by GFP expression) were photographed (under 200X magnification) at 12 or 24 hpi of VSV-GFP, and calculated the virus titer by standard plaque assay with some modifications. To find out the effective time point which MLT exhibits virucidal activity, 2.0 μg/ml of MLT and VSV-GFP (MOI=0.2) were co-incubated for 5, 10, 20, and 30 min at 4°C, and then mixture was inoculated to HEK293T cell monolayers in 6 well plates.

abstract:
  Bee venom (BV) from honey bee (Apis Melifera L.) contains at least 18 pharmacologically active components including melittin (MLT), phospholipase A(2) (PLA(2)), and apamin etc. BV is safe for human treatments dose dependently and proven to possess different healing properties including antibacterial and antiparasitidal properties. Nevertheless, antiviral properties of BV have not well investigated. Hence, we identified the potential antiviral properties of BV and its component against a broad panel of viruses. Co-incubation of non-cytotoxic amounts of BV and MLT, the main component of BV, significantly inhibited the replication of enveloped viruses such as Influenza A virus (PR8), Vesicular Stomatitis Virus (VSV), Respiratory Syncytial Virus (RSV), and Herpes Simplex Virus (HSV). Additionally, BV and MLT also inhibited the replication of non-enveloped viruses such as Enterovirus-71 (EV-71) and Coxsackie Virus (H3). Such antiviral properties were mainly explained by virucidal mechanism. Moreover, MLT protected mice which were challenged with lethal doses of pathogenic influenza A H1N1 viruses. Therefore, these results provides the evidence that BV and MLT could be a potential source as a promising antiviral agent, especially to develop as a broad spectrum antiviral agent.


id: cord-348799-qu4zin3o
author: Wu, Nannan
date: 2019-10-10
txt: ./txt/cord-348799-qu4zin3o.txt
cache: ./cache/cord-348799-qu4zin3o.txt
extension: .txt
mime: text/plain
url: https://doi.org/10.1371/journal.ppat.1008093
doi: 10.1371/journal.ppat.1008093
summary:
  This lack of specificity was not present in cells however, given that the ectopic expression of ISG20 did not lead to the indiscriminate degradation of cellular RNAs (ribosomal as well as small nucleolar RNAs previously shown to be associated to ISG20 [2] ), nor of an Hepatitis C virus (HCV)-derived Luciferase reporter mRNA produced in vitro and then transfected into cells (S4C and S4D Fig) , suggesting that in cells the RNase activity of ISG20 is tightly controlled. Self mimicry allows the escape of target genes'' mRNAs from the effects of ISG20 VSV infection and ectopic DNA or RNA transfection do not bear much in common apart from the fact that both represent artificial injections into the cell of non-self genetic material from without. To further determine whether ISG20 acted on translation initiation and more specifically through specific initiation factors (eIFs), capped and polyadenylated mRNAs coding luciferase under the control of several 5'' untranslated regions (5'' UTRs) were generated in vitro and directly transfected in ISG20-expressing cells (Fig 3D) .

title:
  The interferon stimulated gene 20 protein (ISG20) is an innate defense antiviral factor that discriminates self versus non-self translation

abstract:
  ISG20 is a broad spectrum antiviral protein thought to directly degrade viral RNA. However, this mechanism of inhibition remains controversial. Using the Vesicular Stomatitis Virus (VSV) as a model RNA virus, we show here that ISG20 interferes with viral replication by decreasing protein synthesis in the absence of RNA degradation. Importantly, we demonstrate that ISG20 exerts a translational control over a large panel of non-self RNA substrates including those originating from transfected DNA, while sparing endogenous transcripts. This activity correlates with the protein’s ability to localize in cytoplasmic processing bodies. Finally, these functions are conserved in the ISG20 murine ortholog, whose genetic ablation results in mice with increased susceptibility to viral infection. Overall, our results posit ISG20 as an important defense factor able to discriminate the self/non-self origins of the RNA through translation modulation.


id: cord-353467-wbtzvm4i
title: Functional incorporation of green fluorescent protein into hepatitis B virus envelope particles
author: Lambert, Carsten
date: 2004-12-05
url: https://www.ncbi.nlm.nih.gov/pubmed/15527842/
doi: 10.1016/j.virol.2004.09.031
abstract:
  The envelope of hepatitis B virus (HBV), containing the L, M, and S proteins, is essential for virus entry and maturation. For direct visualization of HBV, we determined whether envelope assembly could accommodate the green fluorescent protein (GFP). While the C-terminal addition of GFP to S trans-dominant negatively inhibited empty envelope particle secretion, the N-terminal GFP fusion to S (GFP.S) was co-integrated into the envelope, giving rise to fluorescent particles. Microscopy and topogenesis analyses demonstrated that the proper intracellular distribution and folding of GFP.S, required for particle export were rescued by interprotein interactions with wild-type S. Thereby, a dual location of GFP, inside and outside the envelope, was observed. GFP.S was also efficiently packaged into the viral envelope, and these GFP-tagged virions retained the capacity for attachment to HBV receptor-positive cells in vitro. Together, GFP-tagged virions should be suitable to monitor HBV uptake and egress in live hepatocytes.


id: cord-354052-x4ckzw64
title: Manipulation of the Porcine Epidemic Diarrhea Virus Genome Using Targeted RNA Recombination
author: Li, Chunhua
date: 2013-08-02
url: https://doi.org/10.1371/journal.pone.0069997
doi: 10.1371/journal.pone.0069997
abstract:
  Porcine epidemic diarrhea virus (PEDV) causes severe economic losses in the swine industry in China and other Asian countries. Infection usually leads to an acute, often lethal diarrhea in piglets. Despite the impact of the disease, no system is yet available to manipulate the viral genome which has severely hampered research on this virus until today. We have established a reverse genetics system for PEDV based on targeted RNA recombination that allows the modification of the 3′-end of the viral genome, which encodes the structural proteins and the ORF3 protein. Using this system, we deleted the ORF3 gene entirely from the viral genome and showed that the ORF3 protein is not essential for replication of the virus in vitro. In addition, we inserted heterologous genes (i.e. the GFP and Renilla luciferase genes) at two positions in the viral genome, either as an extra expression cassette or as a replacement for the ORF3 gene. We demonstrated the expression of both GFP and Renilla luciferase as well as the application of these viruses by establishing a convenient and rapid virus neutralization assay. The new PEDV reverse genetics system will enable functional studies of the structural proteins and the accessory ORF3 protein and will allow the rational design and development of next generation PEDV vaccines.


