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https://www.readbyqxmd.com/read/29163915/field-deployable-quantitative-rapid-identification-of-active-ebola-virus-infection-in-unprocessed-blood
#1
Kavit Shah, Emma Bentley, Adam Tyler, Kevin S R Richards, Edward Wright, Linda Easterbrook, Diane Lee, Claire Cleaver, Louise Usher, Jane E Burton, James K Pitman, Christine B Bruce, David Edge, Martin Lee, Nelson Nazareth, David A Norwood, Sterghios A Moschos
The West African Ebola virus outbreak underlined the importance of delivering mass diagnostic capability outside the clinical or primary care setting in effectively containing public health emergencies caused by infectious disease. Yet, to date, there is no solution for reliably deploying at the point of need the gold standard diagnostic method, real time quantitative reverse transcription polymerase chain reaction (RT-qPCR), in a laboratory infrastructure-free manner. In this proof of principle work, we demonstrate direct performance of RT-qPCR on fresh blood using far-red fluorophores to resolve fluorogenic signal inhibition and controlled, rapid freeze/thawing to achieve viral genome extraction in a single reaction chamber assay...
November 1, 2017: Chemical Science
https://www.readbyqxmd.com/read/29151351/immucillins-in-infectious-diseases
#2
Gary B Evans, Peter C Tyler, Vern L Schramm
Transition state theory proposes that stable chemical mimics of enzymatic transition states will be powerful inhibitors of their cognate enzymes. The Immucillins are chemically stable analogues that mimic the ribocation and leaving-group features of N-ribosyltransferase transition states. Infectious disease agents often rely on ribosyltransferase chemistry in pathways involving precursor synthesis for nucleic acids, salvage of nucleic acid precursors from the host or synthetic pathways with nucleoside intermediates...
November 19, 2017: ACS Infectious Diseases
https://www.readbyqxmd.com/read/29144446/structure-and-assembly-of-the-ebola-virus-nucleocapsid
#3
William Wan, Larissa Kolesnikova, Mairi Clarke, Alexander Koehler, Takeshi Noda, Stephan Becker, John A G Briggs
Ebola and Marburg viruses are filoviruses: filamentous, enveloped viruses that cause haemorrhagic fever. Filoviruses are within the order Mononegavirales, which also includes rabies virus, measles virus, and respiratory syncytial virus. Mononegaviruses have non-segmented, single-stranded negative-sense RNA genomes that are encapsidated by nucleoprotein and other viral proteins to form a helical nucleocapsid. The nucleocapsid acts as a scaffold for virus assembly and as a template for genome transcription and replication...
November 16, 2017: Nature
https://www.readbyqxmd.com/read/29142131/single-dose-trivalent-vesiculovax-vaccine-protects-macaques-from-lethal-ebolavirus-and-marburgvirus-challenge
#4
Demetrius Matassov, Chad E Mire, Theresa Latham, Joan B Geisbert, Rong Xu, Ayuko Ota-Setlik, Krystle N Agans, Dean J Kobs, Morgan Q S Wendling, Amanda Burnaugh, Thomas L Rudge, Carol L Sabourin, Michael A Egan, David K Clarke, Thomas W Geisbert, John H Eldridge
Previous studies demonstrated that a single intramuscular (IM) dose of an attenuated vesicular stomatitis virus vector (Vesiculovax™, rVSV-N4CT1) expressing the glycoprotein (GP) from the Mayinga strain of Zaire ebolavirus (EBOV) protected nonhuman primates (NHP) from lethal challenge with EBOV Kikwit and Makona strains. Here we studied the immunogenicity of an expanded range of attenuated rVSV vectors expressing filovirus GP in mice. Based on data from those studies an optimal attenuated tri-valent rVSV vector formulation was identified which included rVSV vectors expressing EBOV, Sudan ebolavirus (SUDV) or Angola strain of Marburg marburgvirus (MARV) GPs...
November 15, 2017: Journal of Virology
https://www.readbyqxmd.com/read/29128378/ebola-virus-disease-report-from-the-task-force-on-tropical-diseases-by-the-world-federation-of-societies-of-intensive-and-critical-care-medicine
#5
REVIEW
Guy A Richards, Tim Baker, Pravin Amin
Ebola virus is a filovirus that can cause fatal hemorrhagic fever (HF) and five distinct species exist that vary in terms of geographical distribution and virulence. Once the more virulent forms enter the human population, transmission occurs primarily through direct contact with infected body fluids and may result in significant outbreaks. The devastating has been the recent West African outbreak. Clinically, signs and symptoms are similar to those of the other VHFs [4]. The incubation period is 2-21days, followed by fever, headache, myalgia, diarrhoea, vomiting and dehydration; thereafter, there may be recovery or deterioration with collapse, neurological manifestations and bleeding, that can lead to a fatal outcome...
November 3, 2017: Journal of Critical Care
https://www.readbyqxmd.com/read/29128377/viral-hemorrhagic-fever-in-the-tropics-report-from-the-task-force-on-tropical-diseases-by-the-world-federation-of-societies-of-intensive-and-critical-care-medicine
#6
REVIEW
Jorge Luis Hidalgo Marroquin, Guy A Richards, Juan Ignacio Silesky Jiménez, Tim Baker, Pravin Amin
Viral hemorrhagic fevers (VHFs) are a group of illnesses caused by four families of viruses namely Arenaviruses, Filoviruses, Bunyaviruses, and Flaviviruses. Humans are not the natural reservoir for any of these organisms and acquire the disease through vectors from animal reservoirs. In some conditions human to human transmission is possible increasing the risk to healthy individuals in the vicinity, more so to Health Care Workers (HCW). The pathogenesis of VHF, though poorly understood, varies according to the viruses involved...
November 4, 2017: Journal of Critical Care
https://www.readbyqxmd.com/read/29118454/production-and-purification-of-filovirus-glycoproteins-in-insect-and-mammalian-cell-lines
#7
Elizabeth C Clarke, Amanda L Collar, Chunyan Ye, Yíngyún Caì, Eduardo Anaya, Derek Rinaldi, Britney Martinez, Sarah Yarborough, Christine Merle, Manfred Theisen, Jiro Wada, Jens H Kuhn, Steven B Bradfute
Filoviruses are highly virulent pathogens capable of causing severe disease. The glycoproteins of filoviruses are the only virally expressed proteins on the virion surface and are required for receptor binding. As such, they are the main candidate vaccine antigen. Despite their virulence, most filoviruses are not comprehensively characterized, and relatively few commercially produced reagents are available for their study. Here, we describe two methods for production and purification of filovirus glycoproteins in insect and mammalian cell lines...
November 8, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29116224/comparison-of-transcriptomic-platforms-for-analysis-of-whole-blood-from-ebola-infected-cynomolgus-macaques
#8
Emily Speranza, Louis A Altamura, Kirsten Kulcsar, Sandra L Bixler, Cynthia A Rossi, Randal J Schoepp, Elyse Nagle, William Aguilar, Christina E Douglas, Korey L Delp, Timothy D Minogue, Gustavo Palacios, Arthur J Goff, John H Connor
Ebola virus disease (EVD) is a serious illness with mortality rates of 20-90% in various outbreaks. EVD is characterized by robust virus replication and strong host inflammatory response. Analyzing host immune responses has increasingly involved multimodal approaches including transcriptomics to profile gene expression. We studied cynomolgus macaques exposed to Ebola virus Makona via different routes with the intent of comparing RNA-Seq to a NanoString nCounter codeset targeting 769 non-human primate (NHP) genes...
November 7, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29109373/development-of-a-lethal-intranasal-exposure-model-of-ebola-virus-in-the-cynomolgus-macaque
#9
Kendra J Alfson, Laura E Avena, Gabriella Worwa, Ricardo Carrion, Anthony Griffiths
Ebola virus (EBOV) is a filovirus that can cause Ebola virus disease (EVD). No approved vaccines or therapies exist for filovirus infections, despite an urgent need. The development and testing of effective countermeasures against EBOV requires use of animal models and a thorough understanding of how the model aligns with EVD in humans. The majority of published studies report outcomes of parenteral exposures for emulating needle stick transmission. However, based on data from EVD outbreaks, close contact exposures to infected bodily fluid seems to be one of the primary routes of EBOV transmission...
October 29, 2017: Viruses
https://www.readbyqxmd.com/read/29106386/sirna-rescues-nonhuman-primates-from-advanced-marburg-and-ravn-virus-disease
#10
Emily P Thi, Chad E Mire, Amy Ch Lee, Joan B Geisbert, Raul Ursic-Bedoya, Krystle N Agans, Marjorie Robbins, Daniel J Deer, Robert W Cross, Andrew S Kondratowicz, Karla A Fenton, Ian MacLachlan, Thomas W Geisbert
Ebolaviruses and marburgviruses belong to the family Filoviridae and cause high lethality in infected patients. There are currently no licensed filovirus vaccines or antiviral therapies. The development of broad-spectrum therapies against members of the Marburgvirus genus, including Marburg virus (MARV) and Ravn virus (RAVV), is difficult because of substantial sequence variability. RNAi therapeutics offer a potential solution, as identification of conserved target nucleotide sequences may confer activity across marburgvirus variants...
November 6, 2017: Journal of Clinical Investigation
https://www.readbyqxmd.com/read/29040311/putative-endogenous-filovirus-vp35-like-protein-potentially-functions-as-an-ifn-antagonist-but-not-a-polymerase-cofactor
#11
Tatsunari Kondoh, Rashid Manzoor, Naganori Nao, Junki Maruyama, Wakako Furuyama, Hiroko Miyamoto, Asako Shigeno, Makoto Kuroda, Keita Matsuno, Daisuke Fujikura, Masahiro Kajihara, Reiko Yoshida, Manabu Igarashi, Ayato Takada
It has been proposed that some non-retroviral RNA virus genes are integrated into vertebrate genomes. Endogenous filovirus-like elements (EFLs) have been discovered in some mammalian genomes. However, their potential roles in ebolavirus infection are unclear. A filovirus VP35-like element (mlEFL35) is found in the little brown bat (Myotis lucifugus) genome. Putative mlEFL35-derived protein (mlEFL35p) contains nearly full-length amino acid sequences corresponding to ebolavirus VP35. Ebola virus VP35 has been shown to bind double-stranded RNA, leading to inhibition of type I interferon (IFN) production, and is also known as a viral polymerase cofactor that is essential for viral RNA transcription/replication...
2017: PloS One
https://www.readbyqxmd.com/read/29034123/ecological-niche-modeling-for-filoviruses-a-risk-map-for-ebola-and-marburg-virus-disease-outbreaks-in-uganda
#12
Luke Nyakarahuka, Samuel Ayebare, Gladys Mosomtai, Clovice Kankya, Julius Lutwama, Frank Norbert Mwiine, Eystein Skjerve
INTRODUCTION: Uganda has reported eight outbreaks caused by filoviruses between 2000 to 2016, more than any other country in the world. We used species distribution modeling to predict where filovirus outbreaks are likely to occur in Uganda to help in epidemic preparedness and surveillance. METHODS: The MaxEnt software, a machine learning modeling approach that uses presence-only data was used to establish filovirus - environmental relationships. Presence-only data for filovirus outbreaks were collected from the field and online sources...
September 5, 2017: PLoS Currents
https://www.readbyqxmd.com/read/29031163/ebola-virus-requires-phosphatidylinositol-3-5-bisphosphate-production-for-efficient-viral-entry
#13
Shirley Qiu, Anders Leung, Yuxia Bo, Robert A Kozak, Sai Priya Anand, Corina Warkentin, Fabiola D R Salambanga, Jennifer Cui, Gary Kobinger, Darwyn Kobasa, Marceline Côté
For entry, Ebola virus (EBOV) requires the interaction of its viral glycoprotein with the cellular protein Niemann-Pick C1 (NPC1) which resides in late endosomes and lysosomes. How EBOV is trafficked and delivered to NPC1 and whether this is positively regulated during entry remain unclear. Here, we show that the PIKfyve-ArPIKfyve-Sac3 cellular complex, which is involved in the metabolism of phosphatidylinositol (3,5) bisphosphate (PtdIns(3,5)P2), is critical for EBOV infection. Although the expression of all subunits of the complex was required for efficient entry, PIKfyve kinase activity was specifically critical for entry by all pathogenic filoviruses...
October 11, 2017: Virology
https://www.readbyqxmd.com/read/29021793/intracellular-crosslinking-of-filoviral-nucleoproteins-with-xintrabodies-restricts-viral-packaging
#14
Tamarand Lee Darling, Laura Jo Sherwood, Andrew Hayhurst
Viruses assemble large macromolecular repeat structures that become part of the infectious particles or virions. Ribonucleocapsids (RNCs) of negative strand RNA viruses are a prime example where repetition of nucleoprotein (NP) along the genome creates a core polymeric helical scaffold that accommodates other nucleocapsid proteins including viral polymerase. The RNCs are transported through the cytosol for packaging into virions through association with viral matrix proteins at cell membranes. We hypothesized that RNC would be ideal targets for crosslinkers engineered to promote aberrant protein-protein interactions, thereby blocking their orderly transport and packaging...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28986842/testing-experimental-therapies-in-a-guinea-pig-model-for-hemorrhagic-fever
#15
Gary Wong, Yuhai Bi, Gary Kobinger, George F Gao, Xiangguo Qiu
Hemorrhagic fever viruses are among the deadliest pathogens known to humans, and often, licensed medical countermeasures are unavailable to prevent or treat infections. Guinea pigs are a commonly used animal for the preclinical development of any experimental candidates, typically to confirm data generated in mice and as a way to validate and support further testing in nonhuman primates. In this chapter, we use Sudan virus (SUDV), a lethal filovirus closely related to Ebola virus, as an example of the steps required for generating a guinea pig-adapted isolate that is used to test a monoclonal antibody-based therapy against viral hemorrhagic fevers...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28986839/minigenome-systems-for-filoviruses
#16
Thomas Hoenen
Filoviruses are among the most pathogenic viruses known to man, and work with live viruses is restricted to maximum containment laboratories. In order to study individual aspects of the virus life cycle outside of maximum containment laboratories, life cycle modeling systems have been established, which use reporter-encoding miniature versions of the viral genome called minigenomes. With basic minigenome systems viral genome replication and transcription can be studied, whereas more advanced systems also allow us to model other aspects of the virus life cycle outside of a maximum containment laboratory...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28986836/hemorrhagic-fever-virus-budding-studies
#17
Ronald N Harty
Independent expression of the VP40 or Z matrix proteins of filoviruses (marburgviruses and ebolaviruses) and arenaviruses (Lassa fever and Junín), respectively, gives rise to the production and release of virus-like particles (VLPs) that are morphologically identical to infectious virions. We can detect and quantify VLP production and egress in mammalian cells by transient transfection, SDS-PAGE, Western blotting, and live cell imaging techniques such as total internal reflection fluorescence (TIRF) microscopy...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28978704/human-metapneumovirus-induces-formation-of-inclusion-bodies-for-efficient-genome-replication-and-transcription
#18
Nicolás Cifuentes-Muñoz, Jean Branttie, Kerri Beth Slaughter, Rebecca Ellis Dutch
Human metapneumovirus (HMPV) causes significant upper and lower respiratory disease to all age groups worldwide. The virus possesses a negative-sense single-stranded RNA genome of approximately 13.3 Kb encapsidated by multiple copies of the nucleoprotein (N), giving rise to helical nucleocapsids. In addition, copies of the phosphoprotein (P) and the large RNA polymerase (L) decorate the viral nucleocapsids. After viral attachment, endocytosis, and fusion mediated by the viral glycoproteins, HMPV nucleocapsids are released into the cell cytoplasm...
October 4, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28959255/immunological-control-of-viral-infections-in-bats-and-the-emergence-of-viruses-highly-pathogenic-to-humans
#19
Tony Schountz, Michelle L Baker, John Butler, Vincent Munster
Bats are reservoir hosts of many important viruses that cause substantial disease in humans, including coronaviruses, filoviruses, lyssaviruses, and henipaviruses. Other than the lyssaviruses, they do not appear to cause disease in the reservoir bats, thus an explanation for the dichotomous outcomes of infections of humans and bat reservoirs remains to be determined. Bats appear to have a few unusual features that may account for these differences, including evidence of constitutive interferon (IFN) activation and greater combinatorial diversity in immunoglobulin genes that do not undergo substantial affinity maturation...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28931675/comparative-transcriptomics-highlights-the-role-of-the-activator-protein-1-transcription-factor-in-the-host-response-to-ebolavirus
#20
James W Wynne, Shawn Todd, Victoria Boyd, Mary Tachedjian, Reuben Klein, Brian Shiell, Megan Dearnley, Alexander J McAuley, Amanda P Woon, Anthony W Purcell, Glenn A Marsh, Michelle L Baker
Ebolavirus and Marburgvirus comprise two genera of negative-sense single-stranded RNA viruses that cause severe hemorrhagic fevers in humans. Despite considerable research efforts, the molecular events following Ebola virus (EBOV) infection are poorly understood. With the view of identifying host factors that underpin EBOV pathogenesis, we compared the transcriptomes of EBOV-infected human, pig, and bat kidney cells using a transcriptome sequencing (RNA-seq) approach. Despite a significant difference in viral transcription/replication between the cell lines, all cells responded to EBOV infection through a robust induction of extracellular growth factors...
December 1, 2017: Journal of Virology
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