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https://www.readbyqxmd.com/read/28456663/comparative-whole-genome-analysis-of-dengue-virus-serotype-2-strains-differing-in-trans-endothelial-cell-leakage-induction-in-vitro
#1
Sneha Singh, M G Anupriya, Easwaran Sreekumar
The role of genetic differences among dengue virus (DENV) in causing increased microvascular permeability is less explored. In the present study, we compared two closely related DENV serotype-2 strains of Cosmopolitan genotype for their in vitro infectivity phenotype and ability to induce trans-endothelial leakage. We found that these laboratory strains differed significantly in infecting human microvascular endothelial cells (HMEC-1) and hepatocytes (Huh7), two major target cells of DENV in in vivo infections...
April 27, 2017: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/28441781/dengue-virus-non-structural-protein-5
#2
REVIEW
Abbas El Sahili, Julien Lescar
The World Health Organization estimates that the yearly number of dengue cases averages 390 million. This mosquito-borne virus disease is endemic in over 100 countries and will probably continue spreading, given the observed trend in global warming. So far, there is no antiviral drug available against dengue, but a vaccine has been recently marketed. Dengue virus also serves as a prototype for the study of other pathogenic flaviviruses that are emerging, like West Nile virus and Zika virus. Upon viral entry into the host cell and fusion of the viral lipid membrane with the endosomal membrane, the viral RNA is released and expressed as a polyprotein, that is then matured into three structural and seven non-structural (NS) proteins...
April 24, 2017: Viruses
https://www.readbyqxmd.com/read/28411164/high-levels-of-local-inter-and-intra-host-genetic-variation-of-west-nile-virus-and-evidence-of-fine-scale-evolutionary-pressures
#3
Dylan J Ehrbar, Kiet A Ngo, Scott R Campbell, Laura D Kramer, Alexander T Ciota
West Nile virus (WNV; Flaviviridae, Flavivirus) has been endemic in New York State (NYS) since its 1999 introduction, yet prevalence in Culex mosquitoes varies substantially over small spatial and temporal scales. It is unclear if viral genetics plays a role in this variability, as genetic and phenotypic characterization on local scales has generally been lacking. In addition, intrahost diversity of circulating strains have not been fully characterized despite the documented role of minority variants in viral fitness and virulence...
April 12, 2017: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/28388487/characterization-of-virus-specific-vesicles-assembled-by-west-nile-virus-non-structural-proteins
#4
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
#5
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/28261172/the-complement-system-in-flavivirus-infections
#6
Jonas N Conde, Emiliana M Silva, Angela S Barbosa, Ronaldo Mohana-Borges
The incidence of flavivirus infections has increased dramatically in recent decades in tropical and sub-tropical climates worldwide, affecting hundreds of millions of people each year. The Flaviviridae family includes dengue, West Nile, Zika, Japanese encephalitis, and yellow fever viruses that are typically transmitted by mosquitoes or ticks, and cause a wide range of symptoms, such as fever, shock, meningitis, paralysis, birth defects, and death. The flavivirus genome is composed of a single positive-sense RNA molecule encoding a single viral polyprotein...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28214373/improved-detection-of-zika-virus-rna-in-human-and-animal-specimens-by-a-novel-highly-sensitive-and-specific-real-time-rt-pcr-assay-targeting-the-5-untranslated-region-of-zika-virus
#7
Jasper Fuk-Woo Chan, Cyril Chik-Yan Yip, Kah-Meng Tee, Zheng Zhu, Jessica Oi-Ling Tsang, Kenn Ka-Heng Chik, Terance Gi-Wai Tsang, Chris Chung-Sing Chan, Vincent Kwok-Man Poon, Siddharth Sridhar, Feifei Yin, Ivan Fan-Ngai Hung, Sandy Ka-Yee Chau, Anna Jinxia Zhang, Kwok-Hung Chan, Kwok-Yung Yuen
OBJECTIVE AND METHOD: We developed and evaluated five novel real-time RT-PCR assays targeting conserved regions in the 5'-untranslated region (5'-UTR), envelope (E'), non-structural protein 2A (NS2A), NS5 and 3'-UTR of the ZIKV genome. RESULTS: The ZIKV-5'-UTR assay exhibited the lowest in vitro limit of detection (5-10 RNA copies/reaction and 3.0 × 10(-1) plaque-forming units/ml). Compared to the modified version of a widely adopted RT-PCR assay targeting the ZIKV-E gene, the ZIKV-5'-UTR assay showed better sensitivity in human clinical specimens, and representative mouse specimens, including many organs which are known to be involved in human ZIKV infection but difficult to obtain in clinical settings...
February 18, 2017: Tropical Medicine & International Health: TM & IH
https://www.readbyqxmd.com/read/28196656/a-cdna-clone-launched-platform-for-high-yield-production-of-inactivated-zika-vaccine
#8
Yujiao Yang, Chao Shan, Jing Zou, Antonio E Muruato, Diniz Nunes Bruno, Barbosa de Almeida Medeiros Daniele, Pedro F C Vasconcelos, Shannan L Rossi, Scott C Weaver, Xuping Xie, Pei-Yong Shi
A purified inactivated vaccine (PIV) using the Zika virus (ZIKV) Puerto Rico strain PRVABC59 showed efficacy in monkeys, and is currently in a phase I clinical trial. High-yield manufacture of this PIV is essential for its development and vaccine access. Here we report an infectious cDNA clone-launched platform to maximize its yield. A single NS1 protein substitution (K265E) was identified to increase ZIKV replication on Vero cells (a cell line approved for vaccine production) for both Cambodian FSS13025 and Puerto Rico PRVABC59 strains...
March 2017: EBioMedicine
https://www.readbyqxmd.com/read/27940224/the-calmodulin-antagonist-w-7-n-6-aminohexyl-5-chloro-1-naphthalenesulfonamide-hydrochloride-inhibits-denv-infection-in-huh-7-cells
#9
Patricia Bautista-Carbajal, Ruben Soto-Acosta, Antonio H Angel-Ambrocio, Margot Cervantes-Salazar, Circe I Loranca-Vega, Mayra Herrera-Martínez, Rosa M Del Angel
Dengue virus (DENV) replicative cycle occurs in the endoplasmic reticulum where calcium ions play an important role in cell signaling. Calmodulin (CaM) is the primary sensor of intracellular Ca(2+) levels in eukaryotic cells. In this paper, the effect of the calmodulin antagonist W-7 in DENV infection in Huh-7 cells was evaluated. W7 inhibited viral yield, NS1 secretion and viral RNA and protein synthesis. Moreover, luciferase activity, encoded by a DENV replicon, was also reduced. A decrease in the replicative complexes formation was clearly observed in W7 treated cells...
January 15, 2017: Virology
https://www.readbyqxmd.com/read/27746901/illustrating-and-homology-modeling-the-proteins-of-the-zika-virus
#10
Sean Ekins, John Liebler, Bruno J Neves, Warren G Lewis, Megan Coffee, Rachelle Bienstock, Christopher Southan, Carolina H Andrade
The Zika virus (ZIKV) is a flavivirus of the family Flaviviridae, which is similar to dengue virus, yellow fever and West Nile virus. Recent outbreaks in South America, Latin America, the Caribbean and in particular Brazil have led to concern for the spread of the disease and potential to cause Guillain-Barré syndrome and microcephaly. Although ZIKV has been known of for over 60 years there is very little in the way of knowledge of the virus with few publications and no crystal structures. No antivirals have been tested against it either in vitro or in vivo...
2016: F1000Research
https://www.readbyqxmd.com/read/27667714/trim52-inhibits-japanese-encephalitis-virus-replication-by-degrading-the-viral-ns2a
#11
Wenchun Fan, Mengge Wu, Suhong Qian, Yun Zhou, Huanchun Chen, Xiangmin Li, Ping Qian
The members of tripartite-motif containing (TRIM) protein participate in various cellular processes and play an important role in host antiviral function. TRIM proteins exert their antiviral activity either directly by degrading viral proteins through their E3 ligase activity, or indirectly by promoting host innate immunity. This study demonstrated for the first time that TRIM52 is a novel antiviral TRIM protein against Japanese encephalitis virus (JEV) infection. Overexpression of TRIM52 restricted JEV replication in BHK-21 and 293T cells...
September 26, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27665292/a-simple-method-for-developing-an-infectious-cdna-clone-of-japanese-encephalitis-virus
#12
Hao Zheng, Xuchen Zheng, Wu Tong, Fei Liu, Chao Liang, Tao Wang, Fei Gao, Liwei Li, Tongling Shan, Guoxin Li, Guangzhi Tong
Flavivirus cDNA clones frequently demonstrate genetic instability in transformed bacteria, which hampers the construction and manipulation of cDNAs for infectious flaviviruses. In this study, we developed a stable, full-length cDNA clone, pJEHEN, of a GI JEV strain HEN0701 using a medium-copy-number pBR322 vector and propagating cDNA clones at room temperature. The virus vJEHEN recovered from the infectious clone was indistinguishable from the parent virus HEN0701 with respect to plaque morphology, growth kinetics, and virulence characteristics...
February 2017: Virus Genes
https://www.readbyqxmd.com/read/27590716/computational-prediction-and-analysis-of-potential-antigenic-ctl-epitopes-in-zika-virus-a-first-step-towards-vaccine-development
#13
Manas R Dikhit, Md Yousuf Ansari, Vijaymahantesh, Kalyani, Rani Mansuri, Bikash R Sahoo, Budheswar Dehury, Ajay Amit, Roshan K Topno, Ganesh C Sahoo, Vahab Ali, Sanjiva Bimal, Pradeep Das
The Zika virus disease is an Aedes mosquito-borne disease caused by the ZIKA virus. The unavailability of vaccines or proper chemotherapeutic treatment emphasizes the need for the development of preventive and therapeutic vaccines. T cell specific epitopes have been used as vaccine candidates to generate desired immune responses against a variety of viral pathogens. Herein, the immune-informatics approach was used for the screening of potential major histocompatibility complex class I restricted epitopes, which may be competent to generate a cell-mediated immune response in humans...
August 31, 2016: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/27443615/hla-drb1-alleles-are-associated-with-different-magnitudes-of-dengue-virus-specific-cd4-t-cell-responses
#14
Daniela Weiskopf, Michael A Angelo, Alba Grifoni, Patrick H O'Rourke, John Sidney, Sinu Paul, Aruna D De Silva, Elizabeth Phillips, Simon Mallal, Sunil Premawansa, Gayani Premawansa, Ananda Wijewickrama, Bjoern Peters, Alessandro Sette
BACKGROUND: Each year dengue virus (DENV) infects 400 million human but causes symptomatic disease in only a subset of patients, suggesting that host genetic factors may play a role. HLA molecules that restrict T-cell responses are one of the most polymorphic host factors in humans. METHODS: Here we map HLA DRB1-restricted DENV-specific epitopes in individuals previously exposed to DENV, to identify the breadth and specificity of CD4(+) T-cell responses. To investigate whether HLA-specific variations in the magnitude of response might predict associations between dengue outcomes and HLA-DRB1 alleles, we assembled samples from hospitalized patients with known severity of disease...
October 1, 2016: Journal of Infectious Diseases
https://www.readbyqxmd.com/read/27296393/predicting-zika-virus-structural-biology-challenges-and-opportunities-for-intervention
#15
Bryan D Cox, Richard A Stanton, Raymond F Schinazi
BACKGROUND: Zika virus is an emerging crisis as infection is implicated in severe neurological disorders-Guillain-Barré syndrome and fetal microcephaly. There are currently no treatment options available for Zika virus infection. This virus is part of the flavivirus genus and closely related to Dengue Fever Virus, West Nile Virus, and Japanese Encephalitis Virus. Like other flaviviruses, the Zika virus genome encodes three structural proteins (capsid, precursor membrane, and envelope) and seven nonstructural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5)...
August 2015: Antiviral Chemistry & Chemotherapy
https://www.readbyqxmd.com/read/27273223/first-isolation-of-west-nile-virus-from-a-dromedary-camel
#16
Sunitha Joseph, Ulrich Wernery, Jade Ll Teng, Renate Wernery, Yi Huang, Nissy Ag Patteril, Kwok-Hung Chan, Shyna K Elizabeth, Rachel Yy Fan, Susanna Kp Lau, Jörg Kinne, Patrick Cy Woo
Although antibodies against West Nile virus (WNV) have been detected in the sera of dromedaries in the Middle East, North Africa and Spain, no WNV has been isolated or amplified from dromedary or Bactrian camels. In this study, WNV was isolated from Vero cells inoculated with both nasal swab and pooled trachea/lung samples from a dromedary calf in Dubai. Complete-genome sequencing and phylogenetic analysis using the near-whole-genome polyprotein revealed that the virus belonged to lineage 1a. There was no clustering of the present WNV with other WNVs isolated in other parts of the Middle East...
June 8, 2016: Emerging Microbes & Infections
https://www.readbyqxmd.com/read/27169727/population-genomics-of-dengue-virus-serotype-4-insights-into-genetic-structure-and-evolution
#17
Vaishali P Waman, Sunitha Manjari Kasibhatla, Mohan M Kale, Urmila Kulkarni-Kale
The spread of dengue disease has become a global public health concern. Dengue is caused by dengue virus, which is a mosquito-borne arbovirus of the genus Flavivirus, family Flaviviridae. There are four dengue virus serotypes (1-4), each of which is known to trigger mild to severe disease. Dengue virus serotype 4 (DENV-4) has four genotypes and is increasingly being reported to be re-emerging in various parts of the world. Therefore, the population structure and factors shaping the evolution of DENV-4 strains across the world were studied using genome-based population genetic, phylogenetic and selection pressure analysis methods...
August 2016: Archives of Virology
https://www.readbyqxmd.com/read/27053551/transmembrane-domains-of-ns2b-contribute-to-both-viral-rna-replication-and-particle-formation-in-japanese-encephalitis-virus
#18
Xiao-Dan Li, Cheng-Lin Deng, Han-Qing Ye, Hong-Lei Zhang, Qiu-Yan Zhang, Dong-Dong Chen, Pan-Tao Zhang, Pei-Yong Shi, Zhi-Ming Yuan, Bo Zhang
UNLABELLED: Flavivirus nonstructural protein 2B (NS2B) is a transmembrane protein that functions as a cofactor for viral NS3 protease. The cytoplasmic region (amino acids 51 to 95) alone of NS2B is sufficient for NS3 protease activity, whereas the role of transmembrane domains (TMDs) remains obscure. Here, we demonstrate for the first time that flavivirus NS2B plays a critical role in virion assembly. Using Japanese encephalitis virus (JEV) as a model, we performed a systematic mutagenesis at the flavivirus conserved residues within the TMDs of NS2B...
June 15, 2016: Journal of Virology
https://www.readbyqxmd.com/read/26907325/in-vitro-and-in-vivo-evaluation-of-mutations-in-the-ns-region-of-lineage-2-west-nile-virus-associated-with-neuroinvasiveness-in-a-mammalian-model
#19
Katalin Szentpáli-Gavallér, Stephanie M Lim, László Dencső, Krisztián Bányai, Penelope Koraka, Albert D M E Osterhaus, Byron E E Martina, Tamás Bakonyi, Ádám Bálint
West Nile virus (WNV) strains may differ significantly in neuroinvasiveness in vertebrate hosts. In contrast to genetic lineage 1 WNVs, molecular determinants of pathogenic lineage 2 strains have not been experimentally confirmed so far. A full-length infectious clone of a neurovirulent WNV lineage 2 strain (578/10; Central Europe) was generated and amino acid substitutions that have been shown to attenuate lineage 1 WNVs were introduced into the nonstructural proteins (NS1 (P250L), NS2A (A30P), NS3 (P249H) NS4B (P38G, C102S, E249G))...
February 19, 2016: Viruses
https://www.readbyqxmd.com/read/26384477/the-a66g-back-mutation-in-ns2a-of-jev-sa14-14-2-strain-contributes-to-production-of-ns1-protein-and-the-secreted-ns1-can-be-used-for-diagnostic-biomarker-for-virulent-virus-infection
#20
Jingman Wang, Xinfeng Li, Jinyan Gu, Yu Fan, Peng Zhao, Ruibing Cao, Puyan Chen
Japanese encephalitis virus (JEV) is the most common cause of the prevalent encephalitis in Asia-Pacific region and poses a serious risk to public health. Here, we developed a reliable reverse genetics system based on the JEV SA14-14-2 strain to further explore the mechanism for the synthesis of NS1' protein and to investigate the function of NS1' protein during virus infection. NS1' is an additional form of NS1 protein with 52 amino acid carboxy-terminal extension and is expressed by the members of the Japanese encephalitis (JE) serogroup due to the translation frameshift...
December 2015: Infection, Genetics and Evolution
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