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  1 / 7071 MEDLINE  
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[PMID]:29030319
[Au] Autor:Dutta S; Celestine MJ; Khanal S; Huddleston A; Simms C; Arca JF; Mitra A; Heller L; Kraj PJ; Ledizet M; Anderson JF; Neelakanta G; Holder AA; Sultana H
[Ad] Endereço:Department of Biological Sciences, Old Dominion University, Norfolk, VA, USA.
[Ti] Título:Coordination of different ligands to copper(II) and cobalt(III) metal centers enhances Zika virus and dengue virus loads in both arthropod cells and human keratinocytes.
[So] Source:Biochim Biophys Acta;1862(1):40-50, 2018 01.
[Is] ISSN:0006-3002
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Trace elements such as copper and cobalt have been associated with virus-host interactions. However, studies to show the effect of conjugation of copper(II) or cobalt(III) metal centers to thiosemicarbazone ligand(s) derived from either food additives or mosquito repellent such as 2-acetylethiazole or citral, respectively, on Zika virus (ZIKV) or dengue virus (serotype 2; DENV2) infections have not been explored. In this study, we show that four compounds comprising of thiosemicarbazone ligand derived from 2-acetylethiazole viz., (E)-N-ethyl-2-[1-(thiazol-2-yl)ethylidene]hydrazinecarbothioamide (acetylethTSC) (compound 1), a copper(II) complex with acetylethTSC as a ligand (compound 2), a thiosemicarbazone ligand-derived from citral (compound 3) and a cobalt(III) complex with a citral-thiosemicarbazone ligand (compound 4) increased DENV2 and ZIKV replication in both mosquito C6/36 cells and human keratinocytes (HaCaT cells). Treatment of both cell lines with compounds 2 or 4 showed increased dengue viral titers at all three tested doses. Enhanced dengue viral plaque formation was also noted at the tested dose of 100µM, suggesting higher production of infectious viral particles. Treatment with the compounds 2 or 4 enhanced ZIKV and DENV2 RNA levels in HeLa cell line and primary cultures of mouse bone marrow derived dendritic cells. Also, pre- or post treatments with conjugated compounds 2 or 4 showed higher loads of ZIKV or DENV2 envelope (E) protein in HaCaT cells. No changes in loads of E-protein were found in ZIKV-infected C6/36 cells, when compounds were treated after infection. In addition, we tested bis(1,10-phenanthroline)copper(II) chloride ([Cu(phen) ]Cl , (compound 5) and tris(1,10-phenanthroline)cobalt(III) chloride ([Co(phen) ]Cl , (compound 6) that also showed enhanced DENV2 loads. Also, we found that copper(II) chloride dehydrate (CuCl ·2H O) or cobalt(II) chloride hexahydrate (CoCl ·6H O) alone had no effects as "free" cations. Taken together, these findings suggest that use of Cu(II) or Co(III) conjugation to organic compounds, in insect repellents and/or food additives could enhance DENV2/ZIKV loads in human cells and perhaps induce pathogenesis in infected individuals or individuals pre-exposed to such conjugated complexes. IMPORTANCE: Mosquito-borne diseases are of great concern to the mankind. Use of chemicals/repellents against mosquito bites and transmission of microbes has been the topic of interest for many years. Here, we show that thiosemicarbazone ligand(s) derived from 2-acetylethiazole or citral or 1,10-phenanthroline upon conjugation with copper(II) or cobalt(III) metal centers enhances dengue virus (serotype 2; DENV2) and/or Zika virus (ZIKV) infections in mosquito, mouse and human cells. Enhanced ZIKV/DENV2 capsid mRNA or envelope protein loads were evident in mosquito cells and human keratinocytes, when treated with compounds before/after infections. Also, treatment with copper(II) or cobalt(III) conjugated compounds increased viral titers and number of plaque formations. These studies suggest that conjugation of compounds in repellents/essential oils/natural products/food additives with copper(II) or cobalt(III) metal centers may not be safe, especially in tropical and subtropical places, where several dengue infection cases and deaths are reported annually or in places with increased ZIKV caused microcephaly.
[Mh] Termos MeSH primário: Cobalto
Complexos de Coordenação
Cobre
Vírus da Dengue/metabolismo
Queratinócitos/virologia
Carga Viral/efeitos dos fármacos
Zika virus/metabolismo
[Mh] Termos MeSH secundário: Animais
Cercopithecus aethiops
Cobalto/química
Cobalto/farmacologia
Complexos de Coordenação/química
Complexos de Coordenação/farmacologia
Cobre/química
Cobre/farmacologia
Culicidae
Células HeLa
Seres Humanos
Queratinócitos/metabolismo
Queratinócitos/patologia
Células Vero
Proteínas do Envelope Viral
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T; RESEARCH SUPPORT, U.S. GOV'T, NON-P.H.S.
[Nm] Nome de substância:
0 (Coordination Complexes); 0 (Viral Envelope Proteins); 3G0H8C9362 (Cobalt); 789U1901C5 (Copper)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180309
[Lr] Data última revisão:
180309
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171015
[St] Status:MEDLINE


  2 / 7071 MEDLINE  
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[PMID]:28456663
[Au] Autor:Singh S; Anupriya MG; Sreekumar E
[Ad] Endereço:Molecular Virology Laboratory, Rajiv Gandhi Centre for Biotechnology (RGCB), Thycaud P.O., Thiruvananthapuram 695014, Kerala, India.
[Ti] Título:Comparative whole genome analysis of dengue virus serotype-2 strains differing in trans-endothelial cell leakage induction in vitro.
[So] Source:Infect Genet Evol;52:34-43, 2017 Aug.
[Is] ISSN:1567-7257
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo: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. There was a reciprocal correlation in infectivity and vascular leakage induced by these strains, with the less infective strain inducing more trans-endothelial cell leakage in HMEC-1 monolayer upon infection. The cells infected with the strain capable of inducing more permeability were found to secrete more Non-Structural protein (sNS1) into the culture supernatant. A whole genome analysis revealed 37 predicted amino acid changes and changes in the secondary structure of 3' non-translated region between the strains. But none of these changes involved the signal sequence coded by the C-terminal of the Envelope protein and the two glycosylation sites within the NS1 protein critical for its secretion, and the N-terminal NS2A sequence important for surface targeting of NS1. The strain that secreted lower levels of NS1 and caused less leakage had two mutations within the NS1 protein coding region, F103S and T146I that significantly changed amino acid properties. A comparison of the sequences of the two strains with published sequences of various DENV strains known to cause clinically severe dengue identified a number of amino acid changes which could be implicated as possible key genetic differences. Our data supports the earlier observations that the vascular leakage induction potential of DENV strains is linked to the sNS1 levels. The results also indicate that viral genetic determinants, especially the mutations within the NS1 coding region, could affect this critical phenotype of DENV strains.
[Mh] Termos MeSH primário: Vírus da Dengue/fisiologia
Células Endoteliais/virologia
Hepatócitos/virologia
Proteínas não Estruturais Virais/genética
[Mh] Termos MeSH secundário: Regiões 3' não Traduzidas
Animais
Permeabilidade Capilar
Linhagem Celular
Vírus da Dengue/genética
Células Endoteliais/citologia
Variação Genética
Genoma Viral
Hepatócitos/citologia
Seres Humanos
Estrutura Secundária de Proteína
Análise de Sequência de RNA
Proteínas não Estruturais Virais/química
Proteínas não Estruturais Virais/secreção
Replicação Viral/fisiologia
[Pt] Tipo de publicação:COMPARATIVE STUDY; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (3' Untranslated Regions); 0 (NS1 protein, Dengue virus type 2); 0 (Viral Nonstructural Proteins)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180309
[Lr] Data última revisão:
180309
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170501
[St] Status:MEDLINE


  3 / 7071 MEDLINE  
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[PMID]:29385205
[Au] Autor:Di Paola N; Freire CCM; Zanotto PMA
[Ad] Endereço:Laboratory of Molecular Evolution and Bioinformatics, Department of Microbiology, Biomedical Sciences Institute, University of Sao Paulo, Sao Paulo, Brazil.
[Ti] Título:Does adaptation to vertebrate codon usage relate to flavivirus emergence potential?
[So] Source:PLoS One;13(1):e0191652, 2018.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Codon adaptation index (CAI) is a measure of synonymous codon usage biases given a usage reference. Through mutation, selection, and drift, viruses can optimize their replication efficiency and produce more offspring, which could increase the chance of secondary transmission. To evaluate how higher CAI towards the host has been associated with higher viral titers, we explored temporal trends of several historic and extensively sequenced zoonotic flaviviruses and relationships within the genus itself. To showcase evolutionary and epidemiological relationships associated with silent, adaptive synonymous changes of viruses, we used codon usage tables from human housekeeping and antiviral immune genes, as well as tables from arthropod vectors and vertebrate species involved in the flavivirus maintenance cycle. We argue that temporal trends of CAI changes could lead to a better understanding of zoonotic emergences, evolutionary dynamics, and host adaptation. CAI appears to help illustrate historically relevant trends of well-characterized viruses, in different viral species and genetic diversity within a single species. CAI can be a useful tool together with in vivo and in vitro kinetics, phylodynamics, and additional functional genomics studies to better understand species trafficking and viral emergence in a new host.
[Mh] Termos MeSH primário: Códon/genética
Flavivirus/genética
Flavivirus/patogenicidade
[Mh] Termos MeSH secundário: Adaptação Fisiológica
Aedes/genética
Aedes/virologia
Animais
Culex/genética
Culex/virologia
Vírus da Dengue/genética
Vírus da Dengue/patogenicidade
Vírus da Dengue/fisiologia
Evolução Molecular
Flavivirus/fisiologia
Genes Essenciais
Genoma Viral
Interações Hospedeiro-Patógeno/genética
Seres Humanos
Mosquitos Vetores/genética
Mosquitos Vetores/virologia
Filogenia
Vírus do Mosaico do Tabaco/genética
Vírus do Mosaico do Tabaco/patogenicidade
Vírus do Mosaico do Tabaco/fisiologia
Vertebrados/genética
Vertebrados/virologia
Vírus do Nilo Ocidental/genética
Vírus do Nilo Ocidental/patogenicidade
Vírus do Nilo Ocidental/fisiologia
Vírus da Febre Amarela/genética
Vírus da Febre Amarela/patogenicidade
Vírus da Febre Amarela/fisiologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Codon)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180302
[Lr] Data última revisão:
180302
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180201
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0191652


  4 / 7071 MEDLINE  
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[PMID]:29335200
[Au] Autor:Lu D; Liu J; Zhang Y; Liu F; Zeng L; Peng R; Yang L; Ying H; Tang W; Chen W; Zuo J; Tong X; Liu T; Hu Y
[Ad] Endereço:State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Shanghai 201203, China; University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing 100049, China.
[Ti] Título:Discovery and optimization of phthalazinone derivatives as a new class of potent dengue virus inhibitors.
[So] Source:Eur J Med Chem;145:328-337, 2018 Feb 10.
[Is] ISSN:1768-3254
[Cp] País de publicação:France
[La] Idioma:eng
[Ab] Resumo:Using a dengue replicon cell line-based screening, we identified 3-(dimethylamino)propyl(3-((4-(4-fluorophenyl)-1-oxophthalazin-2(1H)-yl)methyl)phenyl)carbamate (10a) as a potent DENV-2 inhibitor, with an IC value of 0.64 µM. A series of novel phthalazinone derivatives based on hit 10a were synthesized and evaluated for their in vitro anti-DENV activity and cytotoxicity. The subsequent SAR study and optimization led to the discovery of the most promising compound 14l, which displayed potent anti-DENV-2 activity, with low IC value against DENV-2 RNA replication of 0.13 µM and high selectivity (SI = 89.2) with acceptable pharmacokinetics profiles.
[Mh] Termos MeSH primário: Antivirais/farmacologia
Vírus da Dengue/efeitos dos fármacos
Descoberta de Drogas
Ftalazinas/farmacologia
[Mh] Termos MeSH secundário: Aedes/citologia
Aedes/virologia
Animais
Antivirais/síntese química
Antivirais/química
Linhagem Celular
Relação Dose-Resposta a Droga
Seres Humanos
Masculino
Camundongos
Testes de Sensibilidade Microbiana
Simulação de Acoplamento Molecular
Estrutura Molecular
Ftalazinas/síntese química
Ftalazinas/química
Relação Estrutura-Atividade
Replicação Viral/efeitos dos fármacos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antiviral Agents); 0 (Phthalazines)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180302
[Lr] Data última revisão:
180302
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180117
[St] Status:MEDLINE


  5 / 7071 MEDLINE  
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[PMID]:27774649
[Au] Autor:Tezuka K; Kuramitsu M; Okuma K; Nojima K; Araki K; Shinohara N; Matsumoto C; Satake M; Takasaki T; Saijo M; Kurane I; Hamaguchi I
[Ad] Endereço:Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
[Ti] Título:Development of a novel dengue virus serotype-specific multiplex real-time reverse transcription-polymerase chain reaction assay for blood screening.
[So] Source:Transfusion;56(12):3094-3100, 2016 12.
[Is] ISSN:1537-2995
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Dengue fever is caused by four related RNA viruses of the genus Flavivirus, dengue virus (DENV)-1, -2, -3, and -4, which are transmitted to humans by mosquitoes. Although DENV is not endemic in Japan, an autochthonous dengue outbreak occurred in 2014. Several transfusion-transmitted cases have also been reported after the use of blood and plasma products in DENV-endemic countries. The aim of this study was to develop a novel multiplex reverse transcription-polymerase chain reaction (RT-PCR) assay for DENV blood screening. STUDY DESIGN AND METHODS: Large-scale oligonucleotide screening was performed to obtain DENV-specific primers and probes using a variety of DENV clinical isolates. A multiplex RT-PCR assay was then developed using the identified oligonucleotides and the ability of this assay to detect DENV RNA was evaluated. RESULTS: A number of oligonucleotides suitable for DENV RNA detection were identified and a novel DENV serotype-specific multiplex RT-PCR assay was successfully established. Comparative analysis revealed that the multiplex assay could detect levels of viral contamination as low as 100 viral copies/mL. CONCLUSION: This established serotype-specific multiplex RT-PCR assay provides a simple, sensitive, and quantitative detection method for DENV, which could be applied in the screening of blood samples to prevent transfusion-transmitted DENV infection.
[Mh] Termos MeSH primário: Vírus da Dengue/genética
Dengue/diagnóstico
Reação em Cadeia da Polimerase/métodos
Sorogrupo
Reação Transfusional
[Mh] Termos MeSH secundário: Segurança do Sangue
Dengue/prevenção & controle
Dengue/transmissão
Seres Humanos
Reação em Cadeia da Polimerase Multiplex
RNA Viral/análise
RNA Viral/sangue
Reação em Cadeia da Polimerase em Tempo Real
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (RNA, Viral)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:180301
[Lr] Data última revisão:
180301
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161028
[St] Status:MEDLINE
[do] DOI:10.1111/trf.13875


  6 / 7071 MEDLINE  
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[PMID]:27770704
[Au] Autor:Wichit S; Ferraris P; Choumet V; Missé D
[Ad] Endereço:Laboratory of MIVEGEC, UMR 224 IRD/CNRS/UM1, Montpellier, France.
[Ti] Título:The effects of mosquito saliva on dengue virus infectivity in humans.
[So] Source:Curr Opin Virol;21:139-145, 2016 12.
[Is] ISSN:1879-6265
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Arboviruses such as Dengue, Chikungunya, and Zika viruses represent a major public health problem due to globalization and propagation of susceptible vectors worldwide. Arthropod vector-derived salivary factors have the capacity to modulate human cells function by enhancing or suppressing viral replication and, therefore, modify the establishment of local and systemic viral infection. Here, we discuss how mosquito saliva may interfere with Dengue virus (DENV) infection in humans. Identification of saliva factors that enhance infectivity will allow the production of vector-based vaccines and therapeutics that would interfere with viral transmission by targeting arthropod saliva components. Understanding the role of salivary proteins in DENV transmission will provide tools to control not only Dengue but also other arboviral diseases transmitted by the same vectors.
[Mh] Termos MeSH primário: Culicidae/virologia
Vírus da Dengue/patogenicidade
Dengue/transmissão
Transmissão de Doença Infecciosa
Interações Hospedeiro-Patógeno
Saliva/metabolismo
[Mh] Termos MeSH secundário: Animais
Seres Humanos
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW; RESEARCH SUPPORT, NON-U.S. GOV'T
[Em] Mês de entrada:1710
[Cu] Atualização por classe:180221
[Lr] Data última revisão:
180221
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161023
[St] Status:MEDLINE


  7 / 7071 MEDLINE  
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[PMID]:29293524
[Au] Autor:Liu WJ; Aaskov JG
[Ad] Endereço:Australian Army Malaria Institute, Weary Dunlop Drive, Enoggera, Brisbane, Australia.
[Ti] Título:Fitness peaks of dengue virus populations.
[So] Source:PLoS One;13(1):e0189554, 2018.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The role of intra-host genetic diversity in dengue viral populations remains a topic of debate, particularly the impact on transmission of changes in this diversity. Several approaches have been taken to increasing and decreasing the genetic diversity of populations of RNA viruses and have drawn what appear to be contradictory conclusions. A 2-6 fold increase in genetic diversity of a wild type population of dengue virus serotype 1(DENV1) and of an infectious clone population derived from the wild type population, produced by treatment with nucleotide analogue 5 fluorouracil (5FU), drove the populations to extinction. Removal of 5FU immediately prior to extinction, resulted in a return to pre-treatment levels of fitness and genetic diversity, albeit with novel single nucleotide polymorphisms. These observations support the concept that DENV populations exist on fitness peaks determined by their transmission requirements and either an increase or a decrease in genetic diversity may result in a loss of fitness.
[Mh] Termos MeSH primário: Vírus da Dengue/fisiologia
[Mh] Termos MeSH secundário: Vírus da Dengue/efeitos dos fármacos
Vírus da Dengue/genética
Fluoruracila/farmacologia
Genes Virais
Variação Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
U3P01618RT (Fluorouracil)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180215
[Lr] Data última revisão:
180215
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180103
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0189554


  8 / 7071 MEDLINE  
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Texto completo SciELO Saúde Pública
[PMID]:28453108
[Au] Autor:Castro-Orozco R; Castro-García LR; Gómez-Camargo DE
[Ad] Endereço:Universidad de San Buenaventura, Cartagena de Indias, Colombia.
[Ti] Título:[Phylogenetic analysis of South American sequences of the nonstructural protein-1 (ns1) of dengue serotype 2 associated with severe clinical bleeding].
[Ti] Título:Análisis filogenético de secuencias suramericanas del gen ns1 del serotipo dengue-2, relacionadas con sangrado clínico severo..
[So] Source:Rev Salud Publica (Bogota);18(3):459-469, 2016 Jun.
[Is] ISSN:0124-0064
[Cp] País de publicação:Colombia
[La] Idioma:spa
[Ab] Resumo:Objective The objective of this in silico study was to compare nucleotide and amino acids DENV-2-NS1 sequences isolated from febrile patients, with and without disease severity, from different South American countries. Matherials and Methods A bayesian MCMC phylogenetic analysis was carried out using 28 complete sequences of the gene NS1 of the DENV-2 serotype (1 056 bp), using MrBayes v.3.2.0 software, with the model SYM+G (2.5 million generations). We also carried out a phylogenetic analysis with Neighbor-Joining method (Jukes-Cantor model). In addition, the amino acids sequences were aligned and compared with each other, using Clustal W included in MEGA v.5.2 software. Results In the amino acids sequences associated with bleeding, the most frequent substitution was isoleucine → threonine at posicion 93. These sequences showed a high percentage (94.6 %) of amino acid homology in comparison with the percentage of amino acids homology (74 %) of DENV-2 isolates not associated with bleeding. Five clades were identified that group the vast majority of the DENV-2-NS1 sequences analyzed (19/24; 79.2 %) with posterior probability values greater than or equal to 58 %. Seven sequences (87.5 %) associated with bleeding were phylogenetically related within clades 4 and 5, the posterior probability values were 58 % and 97 %, respectively. Conclusion Neither phylogenetic characteristics nor differences between amino acids of the DENV-2-NS1 sequences studied were found that could be associated directly with severity of the disease.
[Mh] Termos MeSH primário: Vírus da Dengue/genética
Hemorragia/virologia
Filogenia
Dengue Grave/virologia
Proteínas não Estruturais Virais/genética
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Sequência de Bases
Teorema de Bayes
Seres Humanos
Sorogrupo
Dengue Grave/complicações
Proteínas não Estruturais Virais/classificação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (NS1 protein, Dengue virus type 2); 0 (Viral Nonstructural Proteins)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180215
[Lr] Data última revisão:
180215
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170429
[St] Status:MEDLINE


  9 / 7071 MEDLINE  
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[PMID]:28453962
[Au] Autor:Iswardy E; Tsai TC; Cheng IF; Ho TC; Perng GC; Chang HC
[Ad] Endereço:Institute of Biomedical Engineering, National Cheng Kung University, Tainan 70101, Taiwan; Department of Physics, Syiah Kuala University, Banda Aceh 23111, Indonesia.
[Ti] Título:A bead-based immunofluorescence-assay on a microfluidic dielectrophoresis platform for rapid dengue virus detection.
[So] Source:Biosens Bioelectron;95:174-180, 2017 Sep 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The proof of concept of utilizing a microfluidic dielectrophoresis (DEP) chip was conducted to rapidly detect a dengue virus (DENV) in vitro based on the fluorescence immunosensing. The mechanism of detection was that the DEP force was employed to capture the modified beads (mouse anti-flavivirus monoclonal antibody-coated beads) in the microfluidic chip and the DENV modified with fluorescence label, as the detection target, can be then captured on the modified beads by immunoreaction. The fluorescent signal was then obtained through fluorescence microscopy, and then quantified by ImageJ freeware. The platform can accelerate an immuno-reaction time, in which the on-chip detection time was 5min, and demonstrating an ability for DENV detection as low as 10 PFU/mL. Furthermore, the required volume of DENV samples dramatically reduced, from the commonly used ~50µL to ~15µL, and the chip was reusable (>50x). Overall, this platform provides a rapid detection (5min) of the DENV with a low sample volume, compared to conventional methods. This proof of concept with regard to a microfluidic dielectrophoresis chip thus shows the potential of immunofluorescence based-assay applications to meet diagnostic needs.
[Mh] Termos MeSH primário: Técnicas Biossensoriais
Vírus da Dengue/isolamento & purificação
Dengue/diagnóstico
Microfluídica
[Mh] Termos MeSH secundário: Bioensaio
Dengue/virologia
Vírus da Dengue/genética
Vírus da Dengue/patogenicidade
Seres Humanos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180214
[Lr] Data última revisão:
180214
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170429
[St] Status:MEDLINE


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[PMID]:29261661
[Au] Autor:Raquin V; Merkling SH; Gausson V; Moltini-Conclois I; Frangeul L; Varet H; Dillies MA; Saleh MC; Lambrechts L
[Ad] Endereço:Insect-Virus Interactions Group, Department of Genomes and Genetics, Institut Pasteur, Paris, France.
[Ti] Título:Individual co-variation between viral RNA load and gene expression reveals novel host factors during early dengue virus infection of the Aedes aegypti midgut.
[So] Source:PLoS Negl Trop Dis;11(12):e0006152, 2017 12.
[Is] ISSN:1935-2735
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Dengue virus (DENV) causes more human infections than any other mosquito-borne virus. The current lack of antiviral strategies has prompted genome-wide screens for host genes that are required for DENV infectivity. Earlier transcriptomic studies that identified DENV host factors in the primary vector Aedes aegypti used inbred laboratory colonies and/or pools of mosquitoes that erase individual variation. Here, we performed transcriptome sequencing on individual midguts in a field-derived Ae. aegypti population to identify new candidate host factors modulating DENV replication. We analyzed the transcriptomic data using an approach that accounts for individual co-variation between viral RNA load and gene expression. This approach generates a prediction about the agonist or antagonist effect of candidate genes on DENV replication based on the sign of the correlation between gene expression and viral RNA load. Using this method, we identified 39 candidate genes that went undetected by conventional pairwise comparison of gene expression levels between DENV-infected midguts and uninfected controls. Only four candidate genes were detected by both methods, emphasizing their complementarity. We demonstrated the value of our approach by functional validation of a candidate agonist gene encoding a sterol regulatory element-binding protein (SREBP), which was identified by correlation analysis but not by pairwise comparison. We confirmed that SREBP promotes DENV infection in the midgut by RNAi-mediated gene knockdown in vivo. We suggest that our approach for transcriptomic analysis can empower genome-wide screens for potential agonist or antagonist factors by leveraging inter-individual variation in gene expression. More generally, this method is applicable to a wide range of phenotypic traits displaying inter-individual variation.
[Mh] Termos MeSH primário: Aedes/virologia
Vírus da Dengue/genética
Dengue/virologia
Insetos Vetores/virologia
Proteínas de Ligação a Elemento Regulador de Esterol/genética
Transcriptoma
[Mh] Termos MeSH secundário: Animais
Sistema Digestório/virologia
Feminino
Perfilação da Expressão Gênica
Regulação da Expressão Gênica
Interações Hospedeiro-Patógeno
Seres Humanos
Proteínas de Insetos/genética
Interferência de RNA
RNA Viral/análise
Carga Viral
Replicação Viral
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Insect Proteins); 0 (RNA, Viral); 0 (Sterol Regulatory Element Binding Proteins)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180212
[Lr] Data última revisão:
180212
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171221
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pntd.0006152



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