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https://www.readbyqxmd.com/read/28435072/development-of-a-pseudovirus-based-assay-for-measuring-neutralizing-antibodies-against-coxsackievirus-b5
#1
Pan Chen, Xing Wu, Yao Su, Xiaotian Hao, Qunying Mao, Zhenglun Liang
Coxsackievirus B5 (CV-B5), an important Coxsackie B virus from genus Enteroviruse within the family Picornaviridae, has also been isolated from Hand, Foot, and Mouth Disease (HFMD) patients, and often associated with neurological manifestations. In this study, we found out that Coxsackievirus B3 (CV-B3) replicon RNA could be encapsidated with CV-B5 capsid to assemble infectious CV-B5 pseudovirus. We then utilized this single round infection system of CV-B5 to develop a neutralizing antibody quantification assay...
April 20, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28432094/the-use-of-synthetic-hybrid-strains-to-determine-the-role-of-replicon-3-in-virulence-of-the-burkholderia-cepacia-complex
#2
Kirsty Agnoli, Roman Freitag, Margarida C Gomes, Christian Jenul, Angela Suppiger, Olga Mannweiler, Carmen Frauenknecht, Daniel Janser, Annette C Vergunst, Leo Eberl
The Burkholderia cepacia complex (Bcc) displays a wealth of metabolic diversity with great biotechnological potential, but the utilisation of these bacteria is limited by their opportunistic pathogenicity to humans. The third replicon of the Bcc, megaplasmid pC3 (0.5 to 1.4 Mb, previously chromosome 3), is important for various phenotypes, including virulence, antifungal and proteolytic activities, and the utilization of certain substrates. Approximately half of plasmid pC3 is well-conserved throughout sequenced Bcc members, while the other half is not...
April 21, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/28430437/discovery-of-an-hcv-ns5b-replicase-palm-site-allosteric-inhibitor-bms-929075-advanced-to-phase-1-clinical-studies
#3
Kap-Sun Yeung, Brett R Beno, Kyle Parcella, John A Bender, Katherine A Grant-Young, Andrew Nickel, Prashantha Gunaga, Prakash Anjanappa, Rajesh Onkardas Bora, Kumaravel Selvakumar, Karen Rigat, Ying-Kai Wang, Mengping Liu, Julie A Lemm, Kathy Mosure, Steven Sheriff, Changhong Wan, Mark Witmer, Kevin Kish, Umesh M Hanumegowda, Xiaoliang Zhuo, Yue-Zhong Shu, Dawn Parker, Roy Haskell, Alicia Ng, Qi Gao, Elizabeth Colston, Joseph Raybon, Dennis M Grasela, Kenneth S Santone, Min Gao, Nicholas A Meanwell, Michael W Sinz, Matthew G Soars, Jay O Knipe, Susan B Roberts, John F Kadow
The hepatitis C virus NS5B replicase is a prime target for the development of direct-acting antiviral drugs for the treatment of chronic HCV infection. Inspired by the overlay of bound structures of three structurally distinct NS5B palm site allosteric inhibitors, the high throughput screening hit anthranilic acid 4, the known benzofuran analog 5 and the benzothiadiazine derivative 6, an optimization process utilizing the simple benzofuran template 7 as a starting point for a fragment growing approach was pursued...
April 21, 2017: Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28427827/antiviral-screen-identifies-ev71-inhibitors-and-reveals-camptothecin-target-dna-topoisomerase-1-as-a-novel-ev71-host-factor
#4
Kan Xing Wu, Justin Jang-Hann Chu
Enterovirus 71 (EV71) is one of the causative agents of hand, foot and mouth disease (HFMD) associated with severe neurological disease. EV71's pathogenesis remains poorly understood and the lack of approved antiviral has led to its emergence as a clinically important neurotropic virus. The goals of this study were to: (i) identify novel anti-EV71 compounds that may serve as lead molecules for therapeutics; and (ii) investigate their targets in downstream studies. We screened a 502-compound library of highly purified natural products for anti-EV71 activities in a cell-based immunofluorescence assay that were then confirmed in viral plaque reduction assays...
April 17, 2017: Antiviral Research
https://www.readbyqxmd.com/read/28427794/occurrence-of-a-novel-class-1-integron-harboring-qnrvc4-in-salmonella-rissen
#5
Sirirat Luk-In, Chaiwat Pulsrikarn, Aroon Bangtrakulnonth, Tanittha Chatsuwan, Wanla Kulwichit
We described qnrVC4 in S. Rissen 166ANSS50, a swine isolate, which was detected in the study on quinolone resistance mechanisms of nontyphoidal Salmonella in Thailand. The isolate was found to harbor a ̴17-kb non-conjugative plasmid carrying qnrVC4 within 8.91kb of a novel In4-like class 1 integron (In805). It contained the multi-drug resistance gene cassettes of qnrVC4-qacH4-aacA4-cmlA7-blaOXA-10-aadA1-dfrA14 and unusual 3'-CS of mobC-IS6100. This 1014-bp qnrVC4 cassette included with promoter (PqnrVC4: -35 TTGAGA and -10 TAGTCT) showed high homology with qnrVC4 in superintegron of V...
March 31, 2017: Diagnostic Microbiology and Infectious Disease
https://www.readbyqxmd.com/read/28416600/induction-of-an-ifn-mediated-antiviral-response-by-a-self-amplifying-rna-vaccine-implications-for-vaccine-design
#6
Timothy Pepini, Anne-Marie Pulichino, Thomas Carsillo, Alicia L Carlson, Farid Sari-Sarraf, Katrin Ramsauer, Jason C Debasitis, Giulietta Maruggi, Gillis R Otten, Andrew J Geall, Dong Yu, Jeffrey B Ulmer, Carlo Iavarone
RNA-based vaccines have recently emerged as a promising alternative to the use of DNA-based and viral vector vaccines, in part because of the potential to simplify how vaccines are made and facilitate a rapid response to newly emerging infections. SAM vaccines are based on engineered self-amplifying mRNA (SAM) replicons encoding an Ag, and formulated with a synthetic delivery system, and they induce broad-based immune responses in preclinical animal models. In our study, in vivo imaging shows that after the immunization, SAM Ag expression has an initial gradual increase...
April 17, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28416549/antiviral-activity-and-resistance-analysis-of-ns3-4a-protease-inhibitor-grazoprevir-and-ns5a-inhibitor-elbasvir-in-hepatitis-c-virus-gt4-replicons
#7
Ernest Asante-Appiah, Stephanie Curry, Patricia McMonagle, Paul Ingravallo, Robert Chase, David Nickle, Ping Qiu, Anita Howe, Frederick C Lahser
Although genotype (GT) 4-infected patients represent a minor overall percentage of the global hepatitis C virus (HCV)-infected population, the high prevalence of the genotype in specific geographic regions coupled with substantial sequence diversity makes it an important genotype to study for antiviral drug discovery and development. We evaluated two direct-acting antiviral agents: grazoprevir, an HCV NS3/4A protease inhibitor, and elbasvir, an HCV NS5A inhibitor, in GT4 replicons prior to clinical studies in this genotype...
April 17, 2017: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/28410411/hepatitis-c-virus-impairs-natural-killer-cell-activity-via-viral-serine-protease-ns3
#8
Chang Mo Yang, Joo Chun Yoon, Jeon Han Park, Jae Myun Lee
Hepatitis C virus (HCV) infection is characterized by a high frequency of chronic cases owing to the impairment of innate and adaptive immune responses. The modulation of natural killer (NK) cell functions by HCV leads to an impaired innate immune response. However, the underling mechanisms and roles of HCV proteins in this immune evasion are controversial, especially in the early phase of HCV infection. To investigate the role of HCV nonstructural proteins especially NS3 in the impairment of NK functions, NK cells were isolated from the PBMCs by negative selection...
2017: PloS One
https://www.readbyqxmd.com/read/28405633/genomic-analysis-of-factors-associated-with-low-prevalence-of-antibiotic-resistance-in-extraintestinal-pathogenic-escherichia-coli-sequence-type-95-strains
#9
Craig M Stephens, Sheila Adams-Sapper, Manraj Sekhon, James R Johnson, Lee W Riley
Extraintestinal pathogenic Escherichia coli (ExPEC) strains belonging to multilocus sequence type 95 (ST95) are globally distributed and a common cause of infections in humans and domestic fowl. ST95 isolates generally show a lower prevalence of acquired antimicrobial resistance than other pandemic ExPEC lineages. We took a genomic approach to identify factors that may underlie reduced resistance. We fully assembled genomes for four ST95 isolates representing the four major fimH-based lineages within ST95 and also analyzed draft-level genomes from another 82 ST95 isolates, largely from the western United States...
March 2017: MSphere
https://www.readbyqxmd.com/read/28395182/sensitive-luminescent-reporter-viruses-reveal-appreciable-release-of-hepatitis-c-virus-ns5a-protein-into-the-extracellular-environment
#10
Nicholas S Eyre, Amanda L Aloia, Michael A Joyce, Monrat Chulanetra, D Lorne Tyrrell, Michael R Beard
The HCV NS5A protein is essential for viral RNA replication and virus particle assembly. To study the viral replication cycle and NS5A biology we generated an infectious HCV construct with a NanoLuciferase (NLuc) insertion within NS5A. Surprisingly, beyond its utility as a sensitive reporter of cytoplasmic viral RNA replication, we also observed strong luminescence in cell culture fluids. Further analysis using assembly-defective viruses and subgenomic replicons revealed that infectious virus production was not required for extracellular NS5A-NLuc activity but was associated with enrichment of extracellular NS5A-NLuc in intermediate-density fractions similar to those of exosomes and virus particles...
April 7, 2017: Virology
https://www.readbyqxmd.com/read/28394283/single-round-infectious-particle-antiviral-screening-assays-for-the-japanese-encephalitis-virus
#11
Chien-Yi Lu, Mann-Jen Hour, Ching-Ying Wang, Su-Hua Huang, Wen-Xiang Mu, Yu-Chun Chang, Cheng-Wen Lin
Japanese Encephalitis virus (JEV) is a mosquito-borne flavivirus with a positive-sense single-stranded RNA genome that contains a big open reading frame (ORF) flanked by 5'- and 3'- untranslated regions (UTRs). Nearly 30,000 JE cases with 10,000 deaths are still annually reported in East Asia. Although the JEV genotype III vaccine has been licensed, it elicits a lower protection against other genotypes. Moreover, no effective treatment for a JE case is developed. This study constructed a pBR322-based and cytomegaloviruses (CMV) promoter-driven JEV replicon for the production of JEV single-round infectious particles (SRIPs) in a packaging cell line expressing viral structural proteins...
April 10, 2017: Viruses
https://www.readbyqxmd.com/read/28390955/evidence-that-compatibility-of-closely-related-replicons-in-clostridium-perfringens-depends-on-linkage-to-parmrc-like-partitioning-systems-of-different-subfamilies
#12
Thomas D Watts, Priscilla A Johanesen, Dena Lyras, Julian I Rood, Vicki Adams
Clostridium perfringens produces an extensive repertoire of toxins and extracellular enzymes, many of which are intimately involved in the progression of disease and are encoded by genes on conjugative plasmids. In addition, many C. perfringens strains can carry up to five of these conjugative toxin or antimicrobial resistance plasmids, each of which has a similar 35kb backbone. This conserved backbone includes the tcp conjugation locus and the central control region (CCR), which encodes genes involved in plasmid regulation, replication and partitioning, including a parMRC partitioning locus...
April 6, 2017: Plasmid
https://www.readbyqxmd.com/read/28389536/whole-genome-sequencing-of-drug-resistant-salmonella-enterica-isolated-from-dairy-cattle-and-humans-in-new-york-and-washington-states-reveals-source-and-geographic-associations
#13
Laura M Carroll, Martin Wiedmann, Henk den Bakker, Julie Siler, Steven Warchocki, David Kent, Svetlana Lyalina, Margaret Davis, William Sischo, Thomas Besser, Lorin D Warnick, Richard V Pereira
Multidrug-resistant (MDR) Salmonella enterica can be spread from cattle to humans through direct contact with animals shedding Salmonella, as well as through the food chain, making MDR Salmonella a serious threat to human health. The objective of this study was to use whole-genome sequencing to compare antimicrobial-resistant (AMR) S. Typhimurium, Newport, and Dublin isolated from dairy cattle and humans in Washington state and New York state at a genotypic and phenotypic level. A total of 90 isolates were selected for the study (37 Typhimurium, 32 Newport, 21 Dublin)...
April 7, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/28388487/characterization-of-virus-specific-vesicles-assembled-by-west-nile-virus-non-structural-proteins
#14
Li Yu, Kazuyo Takeda, Yamei Gao
Flavivirus genome encodes seven non-structural proteins (NSPs) and these NSPs are believed to be involved in their genomic RNA replication, of which the mechanism is unclear. We find that West Nile virus (WNV) NSPs were capable of self-assembling membranous vesicles in cells, which are composed of the host endoplasmic reticulum membrane integrated with viral NS1 and NS4A, and possibly NS2A. The vesicles can further organize into replication complex (RC)-associated vesicles which combine both the vesicle and predicted RC components...
June 2017: Virology
https://www.readbyqxmd.com/read/28381578/mutagenesis-of-dengue-virus-protein-ns2a-revealed-a-novel-domain-responsible-for-virus-induced-cytopathic-effect-and-interactions-between-ns2a-and-ns2b-transmembrane-segments
#15
Ren-Huang Wu, Ming-Han Tsai, Kuen-Nan Tsai, Jia Ni Tian, Jian-Sung Wu, Su-Ying Wu, Jyh-Haur Chern, Chun-Hong Chen, Andrew Yueh
The NS2A protein of Dengue virus (DENV) has eight predicted transmembrane segments (pTMS1-8) and participates in RNA replication, virion assembly, and host antiviral response. However, the roles of specific amino acid residues within the pTMS regions of NS2A during the viral life cycle are not clear. Here, we explored the function of DENV NS2A by introducing a series of alanine substitutions into the N-terminal half (pTMS1-4) of the protein in the context of a DENV infectious clone or subgenomic replicon. Six NS2A mutants (NM5, 7, 9, and 17-19) around pTMS1-2 displayed a novel phenotype showing a >1000-fold reduction in virus yield, an absence of plaque formation despite wild-type-like replicon activity, and infectious virus-like particle yields...
April 5, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28380311/spread-of-ctx-type-extended-spectrum-%C3%AE-lactamase-producing-escherichia-coli-isolates-of-epidemic-clone-b2-o25-st131-among-dogs-and-cats-in-japan
#16
Kumiko Kawamura, Tae Sugawara, Nao Matsuo, Kengo Hayashi, Chihiro Norizuki, Kiyoko Tamai, Takaaki Kondo, Yoshichika Arakawa
This study was performed to investigate the carriage rates of CTX-M-type extended-spectrum β-lactamase (ESBL)-producing Escherichia coli among ill companion animals in Japan. Among the 178 nonrepetitive E. coli isolates, including 131 from dogs and 47 from cats, collected between September and November 2015, 42 (23.6%) isolates from 29 dogs and 13 cats were identified as ESBL producers. The antimicrobial susceptibility, O serotype, phylogenetic group, β-lactamase genotype, plasmid replicon type, and sequence type (ST) of each isolate were analyzed...
April 5, 2017: Microbial Drug Resistance: MDR: Mechanisms, Epidemiology, and Disease
https://www.readbyqxmd.com/read/28380016/carbapenemase-producing-enterobacteriaceae-recovered-from-a-spanish-river-ecosystem
#17
Núria Piedra-Carrasco, Anna Fàbrega, William Calero-Cáceres, Thais Cornejo-Sánchez, Maryury Brown-Jaque, Alba Mir-Cros, Maite Muniesa, Juan José González-López
The increasing resistance to carbapenems is an alarming threat in the fight against multiresistant bacteria. The dissemination properties of antimicrobial resistance genes are supported by their detection in a diverse population of bacteria, including strains isolated from the environment. The objective of this study was to investigate the presence of carbapenemase-producing Enterobacteriaceae (CPE) collected from a river ecosystem in the Barcelona metropolitan area (Spain). Identification of β-lactamases and other resistance determinants was determined as was the antimicrobial susceptibility profile...
2017: PloS One
https://www.readbyqxmd.com/read/28379732/prevalence-of-plasmid-mediated-fosfomycin-resistance-gene-fosa3-among-ctx-m-producing-escherichia-coli-isolates-from-chickens-in-china
#18
Wei Jiang, Shuai Men, Linghan Kong, Suzhen Ma, Yongqiang Yang, Yongxiang Wang, Qiwu Yuan, Guangyang Cheng, Wencheng Zou, Hongning Wang
The aim of this study was to investigate the prevalence of fosfomycin resistance gene fosA3 and characterize plasmids harboring fosA3 among CTX-M-producing Escherichia coli from chickens in China. A total of 234 CTX-M-producing E. coli isolates collected from chickens from 2014 to 2016 were screened for the presence of plasmid-mediated fosfomycin resistance genes (fosA, fosA3, and fosC2). Clonal relatedness of fosA3-positive isolates was determined by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST)...
April 2017: Foodborne Pathogens and Disease
https://www.readbyqxmd.com/read/28376860/expansion-of-highly-stable-bla-oxa-10-%C3%AE-lactamase-family-within-diverse-host-range-among-nosocomial-isolates-of-gram-negative-bacilli-within-a-tertiary-referral-hospital-of-northeast-india
#19
Anand Prakash Maurya, Debadatta Dhar, Mridul Kumar Basumatary, Deepjyoti Paul, Birson Ingti, Debarati Choudhury, Anupam Das Talukdar, Atanu Chakravarty, Shweta Mishra, Amitabha Bhattacharjee
BACKGROUND: The current study reports dissemination of highly stable bla OXA-10 family of beta lactamases among diverse group of nosocomial isolates of Gram-negative bacilli within a tertiary referral hospital of the northern part of India. METHODS: In the current study, a total number of 590 Gram negative isolates were selected for a period of 1 year (i.e. 1st November 2011-31st October 2012). Members of Enterobacteriaceae and non fermenting Gram negative rods were obtained from Silchar Medical College and Hospital, Silchar, India...
April 4, 2017: BMC Research Notes
https://www.readbyqxmd.com/read/28374252/alphavirus-based-vaccines
#20
Kenneth Lundstrom
Alphavirus-based vectors have been engineered from Semliki Forest virus, Sindbis virus, and Venezuelan equine encephalitis virus and applied for vaccine development. Immunization in preclinical animal models has been conducted with naked RNA replicons, recombinant viral particles and layered DNA-RNA vectors. Most commonly, the targets for the immunization have been viral surface proteins and tumor antigens, which have elicited strong immune responses and even provided protection against challenges with lethal doses of virus and tumor cells, respectively...
2017: Methods in Molecular Biology
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